Dental plaque forms sequentially, with Fusobacterium nucleatum facilitating the adhesion of pathogenic late colonizers. We hypothesize that a single-chain variable fragment (scFv) antibody library will enable the identification of F. nucleatum adhesins and help elucidate the molecular mechanisms of coaggregation between F. nucleatum and other bacteria. A 4X10^8 clones scFv phage display library was created using spleen RNA from a mouse immunized with F. nucleatum. The library was enriched by biopanning against F. nucleatum 6 times and 292 individual clones tested by ELISA reacted strongly to F. nucleatum. Sixty-two of those clones inhibited F. nucleatum coaggregation with Streptococcus sanguinus. Analysis of select clones revealed differences in coaggregation inhibition, recognition of outer membrane proteins, and BstOI restriction pattern. DNA sequencing showed 6 unique scFvs and of them 3 strongly inhibited interaction with 5 Streptococcus species. These scFvs recognize the outer membrane autotransporter protein RadD (Fn1526), as determined by mass spectrometry. / Farhan Khan placed second in the International Association for Dental Research/Unilever Hatton Competition in the Senior Basic Science Research Category representing Canada, while presenting the research contained in this dissertation. This international competition took place during the 90th General Session & Exhibition of the International Association for Dental Research in Iguaçu Falls, Brazil in June 2012.

Terminal restriction fragment length polymorphism (T-RFLP) analysis and pyrosequencing were used to characterize the microbiota of the tonsil of the soft palate of 126 unfit and 18 healthy pigs. The T-RFLP analysis method was first optimized for the study of the pig tonsil microbiota and the data compared with culture-based identification of common pig pathogens. Putative identifications of the members of the microbiota revealed that the phyla Firmicutes, Proteobacteria and Bacteroidetes were the most prevalent. A comparison of the T-RFLP analysis results grouped into clusters to clinical conditions revealed paleness, abscess, PRRS virus, and Mycoplasma hyopneumoniae to be significantly associated with cluster membership. T-RFLP analysis was also used to select representative tonsil samples for pyrosequencing. These studies confirmed Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Proteobacteria to be the core phyla of the microbiota of the tonsil of the soft palate of pigs. / OMAFRA Animal Health Strategic Investment

Ce travail a porté sur une nouvelle protéine impliquée dans l'apoptose, la mitogaligine, et plus particulièrement sur le fragment interne [31-53] responsable de son adressage à la mitochondrie. L'objectif général du projet est de comprendre au niveau atomique son mécanisme d'action sur les membranes mitochondriales. Le fragment d'adressage est à lui seul cytotoxique. C'est pourquoi j'ai concentré l'essentiel de mon travail de thèse sur son étude. Nous avons défini sa toxicité sur des cellules humaines et montré qu'il perturbait l'intégrité membranaire, excluant certaines protéines de la mitochondrie. Ce phénomène concorde avec le relargage de cytochrome c, à l'origine du déclenchement de l'apoptose par la voie mitochondriale. Pour mieux comprendre le mode d'action du fragment d'adressage et le rôle joué par la cardiolipine, lipide caractéristique des membranes mitochondriales, j'ai étudié par différentes techniques biophysiques complémentaires l'effet du milieu membranaire sur la structuration du peptide et l'effet du peptide sur la membrane. Le peptide a une très forte affinité (13nM) pour des membranes contenant de la cardiolipine. Il se place à plat sur la membrane, s'enfouissant dans l'interface, sans induire d'organisation particulière des lipides. De plus, nous avons mis en évidence que le peptide était capable d'induire une courbure positive de la membrane, ce qui va interférer avec de nombreux processus vitaux pour la cellule. Enfin, pour réaliser les études structurales et fonctionnelles de la protéine entière, j'ai participé aux premières étapes de production de mitogaligine, qui s'est avéré très délicate aussi bien par voie d'expression que par synthèse chimique.

Universiẗat, Diss., 1999--Wuppertal.

Locating a ligand-binding site is an important first step in structure-guided drug discovery, but current methods typically assess the pocket as a whole, doing little to suggest which regions and interactions are the most important for binding. This thesis introduces Fragment Hotspot Maps, a grid-based method that samples atomic propensities derived from interactions in the Cambridge Structural Database (CSD) with simple molecular probes. These maps specifically highlight fragment-binding sites and their corresponding pharmacophores, offering more precision over other binding site prediction methods. The method is validated by scoring the positions of 21 fragment and lead pairs. Fragment atoms are found in the highest scoring parts of the map corresponding to their atom type, with a median percentage rank of 98%. This is reduced to 72% for lead atoms, showing that the method can differentiate between the hotspots, and the warm spots later used during fragment elaboration. For ligand-bound structures, they provide an intuitive visual guide within the binding site, directing medicinal chemists where to grow the molecule and alerting them to suboptimal interactions within the original hit. These calculations are easily accessible through a simple to use web application, which only requires an input PDB structure or code. High scoring specific interactions predicted by the Fragment Hotspot Maps can be used to guide existing computer aided drug discovery methods. The Hotspots Python API has been created to allow these work flows to be executed programmatically through a single Python script. Two of the functions use scores from the Fragment Hotspot Maps to guide virtual screening methods, docking and field-based ligand screening. Docking virtual screening performance is improved by using a constraint selected from the highest scoring polar interaction. The field-based ligand screener uses modified versions of the Fragment Hotspot Maps directly to predict and score the binding pose. This workflow gave comparable results to docking, and for one target, Glucocorticoid receptor (GCR), showed much better results, highlighting its potential as an orthogonal approach. Fragment Hotspot Maps can be used at multiple stages of the drug discovery process, and research into these applications is ongoing. Their utility in the following areas are currently being explored: to assess ligandability for both individual structures and across proteomes, to aid in library design, to assess pockets throughout a molecular dynamics trajectory, to prioritise crystallographic fragment hits and to guide hit-to-lead development.

No description available.

The objective of this work is to analyze the relation of a literary text to one's identity, to its understanding and to its formation through the written language. The relation between man and the world founded on the awareness of one's own visibility serves as the basis for this analysis. Further, the analysis draws on a hypothesis that one's own visibility within the world is perceived as both a threat and as something inauthentic. Thus, human beings make considerable effort to form themselves according to their own principles or to become someone else with the help of literary text, to take on various disguises and masks, but also to use language as a tool enabling them to uncover their own authenticity. On the onset, such approach was represented by Stendhal's work that is considered as the founding stone of literary egotism. The reason is that, in his work, Stendhal focuses exclusively on himself, which is manifested by a double gesture: self-analysis and self-construction. Both these gestures, performed by the means of language, have a number of successors in various literatures, including Czech literature and the work of Jakub Deml. In Deml's texts, too, the double gesture of self-analysis and self-construction can be recognized. By modifying his own identity, mainly through the inclusion of other...

Cette dissertation commence avec les questions et les inquiétudes qui émergent de la lecture de l’oeuvre de Hila Hilst et aussi d'un désir théorique qui traverse mon chemin académique. Quand je mets les textes de l'auteur face au concept de la dispersion de Maurice Blanchot, je n'ai pas l'intention de m'enfermer dans une analyse littéraire dépliée en ses genres, ici ou ailleurs. Où va Hilda Hilst ? Où va la théorie littéraire? À son disparition, comme Blanchot dit en réponse à la question: “Où va la littérature?" Dans ce sens je m'inquiète de m’éloigner du vouloir-saisir des théories, pour que les mots de Hilst apparaissent dans l'état de devenir poétique, comme en fait ils le sont. Alors, les choix que j'ai faits des textes littéraires de Hilst ont suivi deux critères : le plaisir de la lecture et le désir de la dispersion. D’ailleurs je me sers de la possibilité de la pluralité de l'objet littéraire dans la postmodernité pour apporter aux textes des réflexions qui surgissent pas seulement de la littérature mais aussi comme de possibles dépliements de ces textes dans le théâtre et dans les arts plastiques. De cette façon, par l'intersection des théories, étant elles surtout la théorie littéraire et la théorie théâtrale, je cherche d'autres voies pour la compréhension des textes de Hilda Hilst, bien comme de l'objet de la littérature. Sous l'inclination du critique littéraire Roland Barthes, mon écriture est construite à travers les notions d'écriture et de fragment. C'est en raison d'une envie de déplacement que la disposition de ces éléments dans un ensemble devient possible. De l'immensité au geste, la littérature d'Hilst s'échappe d'une représentation et est conduite à ce que je comprends par théâtralité. Ainsi le déplacement est aperçu au long d'un parcours instable et inachevé d'une analyse littéraire et y compris d'un corps qui se promène pour d'autres langages, où il est possible de se retrouver au bord de ce chemin Walter Benjamin. Le « je » qui traverse mon écriture n'es pas effacé, puisque pour que je devienne une autre il est nécessaire détendre ce « je » à travers des langages qui le constituent. Parmi les divers faits et gestes de la littérature comparée, rien ne m'intéresse autant que les traces d'une liberté qui est là offerte au domaine théorique de la littérature, même devant beaucoup d'impasses. La liberté qui est ici assumée dans son écriture et dans son responsabilité. / Esta dissertação parte dos questionamentos e das inquietudes que surgem da leitura da obra de Hilda Hilst e de um pulsar teórico que transpassa minha trajetória acadêmica. Uma vez que coloco os textos da autora diante do conceito de dispersão de Maurice Blanchot, não intento me delimitar a uma análise literária desdobrada em seus gêneros, nesse ou naquele lugar. Para onde vai Hilda Hilst? Para onde vai a teoria literária? Ao seu desaparecimento, como diz Blanchot em resposta à pergunta: “Para onde vai a literatura?” Nesse sentido, preocupo-me em distanciar-me do querer-agarrar das teorias, para que as palavras de Hilst apareçam em estado de devir poético, como de fato elas são. Sendo assim, as escolhas que fiz dos textos literários de Hilst levaram em conta dois critérios: o prazer na leitura e o desejo de dispersão. Além disso, utilizo-me da possibilidade de pluralidade do objeto literário na pós-modernidade para trazer ao texto reflexões que surgem não só da literatura, mas também de seus possíveis desdobramentos no teatro e nas artes plásticas. De tal modo, através do entrecruzamento de teorias, sendo elas, principalmente, a literária e a teatral, busco outras vias à compreensão dos textos literários da autora, bem como do objeto da literatura. Sob o viés da crítica literária de Roland Barthes, minha escrita é construída por meio dos conceitos de escritura e fragmento. É devido a um anseio de deslocamento que a disposição desses elementos em conjunto torna-se possível. Da imensidão ao gesto, a literatura hilstiana escapa de uma representação e é conduzida ao que entendo por teatralidade. Assim, o deslocamento é percebido no percurso instável e inconcluso de uma análise literária e, inclusive, de um corpo que passeia por outras linguagens, onde Walter Benjamin aparece à margem desse caminho. O eu que atravessa minha escrita não é obliterado, já que, para tornar-me outra, é necessário distender esse eu através das linguagens que o constituem. Entre os diversos fazeres da literatura comparada, nada me interessa tanto quanto os rastros de uma liberdade que ali é disposta ao teórico da literatura, mesmo que diante de tantos impasses. Liberdade que é aqui assumida em sua escrita e também em sua responsabilidade.

Orientador: Vera Lex Engel / Banca: Sebastião Venâncio Martins / Banca: Renata Cristina Batista Fonseca / Resumo: A Mata Atlântica é um dos biomas de maior biodiversidade do planeta, que há séculos tem sido sistematicamente destruída pelo desmatamento e fragmentação florestal, principalmente no domínio da Floresta Estacional Semidecidual. Os fragmentos remanescentes frequentemente têm sofrido distúrbios adicionais, tanto antrópicos quanto naturais. Deste modo, para a sua conservação e restauração é fundamental o conhecimento da resposta desses ecossistemas aos distúrbios. Este trabalho teve como objetivo geral caracterizar a estrutura da vegetação e a regeneração natural de um fragmento de floresta estacional semidecidual com diferentes históricos de perturbações e conhecer o papel das clareiras na regeneração de algumas das suas espécies arbóreas mais representativas. A pesquisa foi realizada em um fragmento de floresta estacional semidecidual com aproximadamente 615 ha (Reserva Particular do Patrimônio Natural (RPPN) Olavo Egydio Setúbal), no município de Lençóis Paulista (22. 27'S, 48.57W), SP. A amostragem foi feita através da demarcação de parcelas permanentes contíguas (10 x 10 m), totalizando um hectare em cada uma de três área na floresta( três hectares no total), sendo: área A, local que sofreu efeitos de um tornado ocorrido há cinco anos atrás; área B: área mais bem conservada, sem histórico recente de perturbações antrópicas e, área C, com registros de extração seletiva de madeira até meados da década de 70. Em cada área foram sorteadas 50 parcelas de 10 x 10 m para o levantamento fitossociológico do estrato arbóreo (indivíduos com altura 1,30 m) e regenerante (altura 0,50 e <1,30 m, levantado em 3 sub-parcelas de 10 x 1 m). Para a regeneração natural, cada subparcela amostrada foi categorizada como sendo de clareira (definição de Brokaw, 1982) ou de dossel fechado. O monitoramento do microclima foi feito quatro vezes... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The Atlantic Forest is one of the richest planest's biomes in biodiversity, which has been systematically destroyed by deforestation and forest fragmentation., mainly at the Seasonal Semideciduous Forest domain. The remaining forest fragments have often suffered further disturbances, be natural or anthropogenic. To conserve or restore these ecosystems is fundamental to know how do they respond to the disturbances. This paper has the objective of characterizing the vegetation structure and the natural regeneration of a forest fragment with different disturbances histories, assessing the role of gaps in the regeneration of its most representatives tree species..The research was realized in a 615 ha seasonal semideciduous forest fragment (Private Natural Patrimony Reserve-RPPN "Olavo Egydio Setúbal"), at Lençóis Paulista Municipality (22. 27'S, 48.57W), SP. Sampling method was by contiguous permanent plots (10 x 10m), totalizing one hectare in each of three study sites within the forest (three ha in the total: Area A: site subjected to a tornado disturbance five years ago; Area B: site well conserved, with no recent anthropogenic disturbances events and, Area C: site were selective logging was registered until meddle 70's. In each site 50 plots (10 x 10 m) were randomly assorted for the phytosociological (stems 1.30 height) and natural regeneration (1.30 < height 0.50 m, surveyed in three 1 x 10 m sub-plots within each plot) assessment. Each sub-plot established for natural regeneration survey was categorized as gap (Brokaw's definition, 1982) or closed canopy. Microclimate was monitored four times in the year, within three consecutive days, in pairs of gap and closed canopy plots, in each of the three study sites. The preference of some tree species for regenerating in gap or closed canopy habitats was also tested. Within the three samples sites we sampled 10.570 individuals... (Complete abstract click electronic access below) / Mestre

Esta pesquisa discute a obra de Jorge Guinle (1947-1987) e o diálogo que o artista travou com a pintura moderna e contemporânea brasileira e internacional. Argumenta que a pintura de Jorge Guinle, embora concentrada apenas nos últimos sete anos da vida do artista, estabeleceu um diálogo profundo com a arte do século XX, sem, com isso, deixar de se lançar a uma indagação de seu próprio tempo. / This research discusses the work of Jorge Guinle (1947-1987) and the dialogue the artist sustained with the Brazilian and international modern and contemporary painting. It argues that Jorge Guinle\'s painting, although concentrated only in the last seven years of the artist\'s life, established a profound dialogue with the art of the twentieth century, without failing, however, to undertake an investigation of its own time.