Suprimento de Nitrogênio para Culturas de Verão pela Aplicação Antecipada em Azevém Pastejado por Ovinos

RESUMO Busca-se nos Sistemas Integrados de Produção Agropecuária (SIPA) maior ciclagem do nitrogênio (N). Este trabalho objetivou avaliar a adubação antecipada em azevém pastejado por ovinos no índice de nutrição nitrogenada (INN) das culturas de verão em SIPA. O estudo foi realizado em um protocolo de longa duração manejado, no período de inverno, sob dois métodos de pastoreio, contínuo e rotativo, e duas intensidades de pastejo, moderada e baixa, com quatro repetições. No verão, a área foi subdividida em dois sistemas de cultivos, soja e rotação soja/milho. A fertilização foi feita na fase pastagem, com 75 kg de N e 60 kg de P2O5 e K2O ha-1. Avaliaram-se a massa de forragem residual (MFR) da pastagem e o rendimento, o teor de N e o INN da fitomassa das culturas de verão. No milho, houve efeito das intensidades de pastejo para INN, ao contrário da soja. Não houve efeito dos métodos de pastoreio. A MFR é importante fonte de N para a cultura de verão subsequente. Menores intensidades de pastejo geraram maiores MFR e INN para a cultura do milho. A adubação antecipada não influenciou a soja, pois essa atende parte da sua demanda por N pela fixação biológica.

Molecular insights into T cell activation : study of the TRAF, Bcl10 and Carma 1 proteins

Abstract : Activation of naïve T lymphocytes is essential for the onset of an adaptive immune response against a pathogenic threat. T lymphocytes are activated through the engagement of their highly specific cell surface antigen-receptor (TCR), together with co-stimulatory receptors, by activated antigen-presenting cells that display antigenic peptide fragments from the pathogen that they have detected. Dissection of the mechanisms that modulate TCR- and co-stimulation- induced signals is therefore crucial for the understanding of the molelcular basis of adaptive immune responses. Following antigen-receptor triggering, the Carma1, Bcl10 and Malt1 (CBM) proteins assemble into an oligomeric complex, which is essential for activation of the NF-κB and JNK signaling pathways in lymphocytes. In this work, by using human epithelial and lymphocytic cell lines, we identified the TNF-receptor-associated factor (TRAF) proteins TRAF3 and TRAF7 as new binding partners of Bcl10 and Carma1, respectively. We could show that TRAF3 is required for the proper transcriptional upregulation of IL-2 in activated T cells, and that endogenous TRAF3 is recruited to Bcl10 following TCR engagement. Although the mechanisms used by TRAF3 to modulate the transcriptional activation of the IL-2 promoter are not elucidated, the stimulus-dependent association ofTRAF3 with its direct binding partner Bcl10 suggests that TRAF3 is regulating Bcl10 function in TCR-activated lymphocytes. We also demonstrated that TRAF7 acts as a negative regulator of Carma1-induced NFκB-and AP1-dependent transcription by overexpression in 293T cells. These data suggest that TRAF7 could contribute to the negative regulation of TCR-dependent Carma1 functions. Finally, we showed that Carma1 is processed upon antigen-receptor triggering in B and T cell lines, as well as in primary human CTLs, and that this processing is dependent on the proteolytic activity of Malt1. Collectively, this work contributes to describe new proteins and regulatory mechanisms that modulate CBM-dependent functions in activated lymphocytes. Furthermore, it uncovers new tracks that could lead to a better molecular understanding of the complex interplay between the activatory and inhibitory regulators associated with the CBM complex. Résumé : L'activation des lymphocytes T naifs est une étape essentielle à la mise en place d'une réponse immunitaire adaptative pour combattre une infection. Après la détection d'un pathogène, les cellules présentatrices d'antigènes exposent à leur surface des fragments peptidiques provenant du pathogène, qui activent le récepteur à antigène (TCR) spécifique des lymphocytes T, ainsi que des molécules co-stimulatrices qui contribuent à l'activation complète des lymphocytes T. La caractérisation des mécanismes qui modulent les cascades de signaux émanant du TCR et des récepteurs de co-stimulation est essentielle à la compréhension du fonctionnement moléculaire de la réponse immunitaire adaptative. La ligation du TCR induit la formation d'un complexe oligomérique comprenant les protéines Carma1, Bcl10 et Malt1, qui est essentiel à l'activation des voies de signalisation cellulaires NF-κB et JNK induisant l'activation complète des lymphorctes T. Dans cette étude, à l'aide de lignées de cellules humaines épithéliales et lymphocytaires, nous avons identifié que deux protéines de la famille des TRAF (Tumor Necrosis Factor Receptor-Associated Factor), TRAF3 et TRAF7, s'associent à Bc110 et à Carma1, respectivement. Les TRAFs sont d'importants régulateurs des voies de signalisation dans les cellules du système immunitaire inné et adaptatif. Nous avons démontré que TRAF3 était important pour permettre la transcription de l'interleukine-2 (IL-2) dans les lymphocytes T activés, et que TRAF3 s'associait à Bc110 à la suite de la stimulation du TCR Les mécanismes que TRAF3 utilise pour moduler l'activation du promoteur de l'IL-2 ne sont pas connus, mais l'association de TRAF3 à Bc110 suite à la stimulation du TCR suggère que TRAF3 régule la fonction de Bc110. Nous avons également identifié TRAF7 comme un nouveau régulateur négatif des voies NF-κB et JNK induites par surexpression de la protéine Carma1. Nos données suggèrent que TRAF7 pourrait également contribuer à la régulation négative de la fonction de Carma1 dans les lymphocytes activés. Enfin, nous avons découvert que Carma1 était clivé suite à la stimulation du TCR, et que ce clivage dépendait de l'activité protéolytique de Malt1. Cette étude contribue ainsi à la description de nouvelles protéines et de nouveaux mécanismes qui modulent l'activité du complexe CBM dans les lymphocytes activés, et ouvre la voie à la caractérisation moléculaire de ces nouveaux mécanismes importants pour la régulation de la réponse immunitaire adaptative.

Persistent infection of arenaviruses : molecular mechanisms and pathogenesis

Arenaviruses are enveloped negative single strand RNA viruses that include a number of important human pathogens. The most prevalent human pathogen among the arenaviruses is the Old World arenavirus Lassa virus (LASV) which is endemic in West Africa from Senegal to Cameroon. LASV is the etiologic agent of a severe viral hemorrhagic fever named Lassa fever whose mortality rate can reach 30% in hospitalized patients. One of the hallmarks of fatal arenavirus infection in humans is the absence of an effective innate and adaptive immune response. In nature, arenaviruses are carried by rodents which represent the natural reservoirs as well as the vectors for transmission. In their natural rodent reservoir, arenaviruses have the ability to establish persistent infection without any overt signs and symptoms of pathology. We believe that the modulation of the host cell's innate immunity by arenaviruses is a key determinant for persistence in the natural host and for the pathogenesis in man. In this thesis, we studied the interaction of arenaviruses with two main branches of the host's innate anti-viral defense, the type I interferon (IFN) system and virus-induced mitochondrial apoptosis. The arenavirus nucleoprotein (NP) is responsible for the anti-IFN activity of arenaviruses. Specifically, NP blocks the activation and the nuclear translocation of the transcription factor interferon regulatory factor 3 (IRF3) which leads to type I IFN production. LASV and the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) NPs contain a 3'-5'exoribonuclease domain in the C terminal part that has been linked to the anti-IFN activity of NP. In the first project, we sought to identify cellular component(s) of the type I IFN induction pathway targeted by the viral NP. Our study revealed that LCMV NP prevents the activation of IRF3 by blocking phosphorylation of the transcription factor. We found that LCMV NP specifically targets the IRF-activating kinase IKKs, and this specific binding is conserved within the Arenaviridae. We could also demonstrate that LCMV NP associates with the kinase domain of IKKs involving NP's C-terminal region. Lastly, we showed that the binding of LCMV NP inhibits the kinase activity of IKKs. This study allowed the discovery of a new cellular interacting partner of arenavirus NP. This newly described association may play a role in the anti-IFN activity of arenaviruses but potentially also in other aspects of arenavirus infection. For the second project, we investigated the ability of arenaviruses to avoid and/or suppress mitochondrial apoptosis. As persistent viruses, arenaviruses evolved a "hit and stay" survival strategy where the apoptosis of the host cell would be deleterious. We found that LCMV does not induce mitochondrial apoptosis at any time during infection. Specifically, no caspase activity, no cytochrome c release from the mitochondria as well as no cleavage of poly (ADP-ribose) polymerase (PARP) were detected during LCMV infection. Interestingly, we found that virus-induced mitochondrial apoptosis remains fully functional in LCMV infected cells, while the induction of type IIFN is blocked. Since both type IIFN production and virus- induced mitochondrial apoptosis critically depend on the pattern recognition receptor (PRR) RIG-I, we examined the role of RIG-I in apoptosis in LCMV infected cells. Notably, virus- induced mitochondrial apoptosis in LCMV infected cells was found to be independent of RIG- I and MDA5, but still depended on MAVS. Our study uncovered a novel mechanism by which arenaviruses alter the host cell's pro-apoptotic signaling pathway. This might represent a strategy arenaviruses developed to maintain this branch of the innate anti-viral defense in absence of type I IFN response. Taken together, these results allow a better understanding of the interaction of arenaviruses with the host cell's innate immunity, contributing to our knowledge about pathogenic properties of these important viruses. A better comprehension of arenavirus virulence may open new avenues for vaccine development and may suggest new antiviral targets for therapeutic intervention against arenavirus infections. - Les arenavirus sont des virus enveloppés à ARN simple brin qui comportent un grand nombre de pathogènes humains. Le pathogène humain le plus important parmi les arenavirus est le virus de Lassa qui est endémique en Afrique de l'Ouest, du Sénégal au Cameroun. Le virus de Lassa est l'agent étiologique d'une fièvre hémorragique sévère appelée fièvre de Lassa, et dont le taux de mortalité peut atteindre 30% chez les patients hospitalisés. L'une des caractéristiques principales des infections fatales à arenavirus chez l'Homme est l'absence de réponse immunitaire innée et adaptative. Dans la nature, les arenavirus sont hébergés par différentes espèces de rongeur, qui représentent à la fois les réservoirs naturels et les vecteurs de transmission des arenavirus. Dans leur hôte naturel, les arenavirus ont la capacité d'établir une infection persistante sans symptôme manifeste d'une quelconque pathologie. Nous pensons que la modulation de système immunitaire inné de la cellule hôte par les arenavirus est un paramètre clé pour la persistance au sein de l'hôte naturel, ainsi que pour la pathogenèse chez l'Homme. L'objectif de cette thèse était d'étudier l'interaction des arenavirus avec deux branches essentielles de la défense antivirale innée de la cellule hôte, le système interféron (IFN) de type I et l'apoptose. La nucléoprotéine virale (NP) est responsable de l'activité anti-IFN des arenavirus. Plus spécifiquement, la NP bloque 1'activation et la translocation nucléaire du facteur de transcription IRF3 qui conduit à la production des IFNs de type I. La NP du virus de Lassa et celle du virus de la chorioméningite lymphocytaire (LCMV), l'arénavirus prototypique, possèdent dans leur extrémité C-terminale un domaine 3'-5' exoribonucléase qui a été associé à l'activité anti-IFN de ces protéines. Dans un premier projet, nous avons cherché à identifier des composants cellulaires de la cascade de signalisation induisant la production d'IFNs de type I qui pourraient être ciblés par la NP virale. Nos recherches ont révélé que la NP de LCMV empêche 1'activation d'IRF3 en bloquant la phosphorylation du facteur de transcription. Nous avons découvert que la NP de LCMV cible spécifiquement la kinase IKKe, et que cette interaction spécifique est conservée à travers la famille des Arenaviridae. Notre étude a aussi permis de démontrer que la NP de LCMV interagit avec le domaine kinase d'IKKe et que l'extrémité C-terminale de la NP est impliquée. Pour finir, nous avons pu établir que l'association avec la NP de LCMV inhibe l'activité kinase d'IKKe. Cette première étude présente la découverte d'un nouveau facteur cellulaire d'interaction avec la NP des arenavirus. Cette association pourrait jouer un rôle dans l'activité anti-IFN des arénavirus, mais aussi potentiellement dans d'autres aspects des infections à arénavirus. Pour le second projet, nous nous sommes intéressés à la capacité des arénavirus à éviter et/ou supprimer l'apoptose mitochondriale. En tant que virus persistants, les arénavirus ont évolué vers une stratégie de survie "hit and stay" pour laquelle l'apoptose de la cellule hôte serait néfaste. Nous avons observé qu'à aucun moment durant l'infection LCMV n'induit l'apoptose mitochondriale. Spécifiquement, aucune activité de caspase, aucune libération mitochondriale de cytochrome c ainsi qu'aucun clivage de la polymerase poly(ADP-ribose) (PARP) n'a été détecté pendant l'infection à LCMV. Il est intéressant de noter que l'apoptose mitochondriale induite par les virus reste parfaitement fonctionnelle dans les cellules infectées par LCMV, alors que l'induction de la réponse IFN de type I est bloquée dans les mêmes cellules. La production des IFNs de type I et l'apoptose mitochondriale induite par les virus dépendent toutes deux du récepteur de reconnaissance de motifs moléculaires RIG-I. Nous avons, par conséquent, investigué le rôle de RIG-I dans l'apoptose qui a lieu dans les cellules infectées par LCMV lorsqu'on les surinfecte avec un autre virus pro-apoptotique. En particulier, l'apoptose mitochondriale induite par les surinfections s'est révélée indépendante de RIG-I et MDA5, mais dépendante de MAVS dans les cellules précédemment infectées par LCMV. Notre étude démontre ainsi l'existence d'un nouveau mécanisme par lequel les arénavirus altèrent la cascade de signalisation pro-apoptotique de la cellule hôte. Il est possible que les arénavirus aient développé une stratégie permettant de maintenir fonctionnelle cette branche de la défense antivirale innée en l'absence de réponse IFN de type I. En conclusion, ces résultats nous amènent à mieux comprendre l'interaction des arénavirus avec l'immunité innée de la cellule hôte, ce qui contribue aussi à améliorer notre connaissance des propriétés pathogéniques de ces virus. Une meilleure compréhension des facteurs de virulence des arénavirus permet, d'une part, le développement de vaccins et peut, d'autre part, servir de base pour la découverte de nouvelles cibles thérapeutiques utilisées dans le traitement des infections à arénavirus.

Protection from lethal Gram-negative bacterial sepsis by targeting Toll-like receptor 4

Rapport de synthèse : L'immunité innée regroupe les mécanismes moléculaires et cellulaires formant la première ligne de défense contre les infections microbiennes. La détection des micro-organismes pathogènes est assurée par des cellules sentinelles (cellules dendritiques et macrophages) qui jouent un rôle fondamental dans l'initiation des mécanismes de défense de l'hôte. Au contact de produits microbiens, ces cellules produisent un large échantillonnage de molécules, dont des cytokines, impliquées dans le développement de la réponse inflammatoire. La régulation de cette réponse relève d'un équilibre délicat, son insuffisance tant que son excès pouvant compromettre le devenir des patients infectés. La sepsis sévère et le choc septique représentent les formes les plus sévères d'infection, et leur mortalité demeure élevée (25 à 30% pour la sepsis sévère et 50 à 60% pour le choc septique). De plus, l'incidence de la sepsis tend à augmenter, atteignant en 2000 plus de 240 cas pour 100'000 personnes en Grande-Bretagne. La sepsis est caractérisée dans sa phase aiguë par une réponse inflammatoire exubérante. La plupart des thérapies visant à la bloquer ont toutefois montré des bénéfices incertains lors de leur application clinique. Il est donc impératif d'identifier de nouvelles cibles thérapeutiques. Les "Toll-like receptors" (TLRs) sont une famille de récepteurs qui jouent un rôle fondamental dans la détection des micro-organismes par les cellules du système immunitaire inné. Parmi eux, TLR4 est indispensable à la reconnaissance du lipopolysaccharide (LPS) des bactéries Gram-négatives. L'interaction entre TLR4 et le LPS représentant un élément précoce de la réponse de l'hôte à l'infection, nous avons émit l'hypothèse que TLR4 pourrait représenter une cible de choix en vue du développement de nouvelles thérapies contre la sepsis. Dans l'objectif de valider ce concept, nous avons, dans un premier temps, démontré que des souris génétiquement déficientes en TLR4 étaient totalement résistantes au choc septique induit par Escherichia coli (E. coli), une bactérie Gram-négative fréquemment responsable de sepsis. Forts de cette observation, nous avons développé une molécule recombinante composée du domaine extracellulaire de TLR4 fusionné à la partie IgGi-Fc. Cette molécule soluble, qui inhibait la réponse des macrophages au LPS in vitro, a été utilisée pour générer des anticorps anti-TLR4 chez le lapin. La spécificité et l'efficacité de ces anticorps ont été prouvées en démontrant que les anti-TLR4 bloquaient les signaux d'activation intracellulaire et la production de TNF et d'IL-6 en réponse au LPS et aux bactéries Gram-négatives in vitro et in vivo. Enfin, l'efficacité des ces anticorps a été testée dans des modèles de sepsis chez la souris. Ainsi, l'injection prophylactique (-lh) ou thérapeutique (+3h) d'anticorps anti-TLR4 réduisait la production de TNF et protégeait les animaux de la mort. De manière spectaculaire, ces anticorps réduisaient également la production de TNF et protégeaient de la sepsis à E. coli lorsqu'ils étaient administrés de manière prophylactique (-4h) et thérapeutique, jusqu'à 13 heures après l'initiation de l'infection. Ces résultats indiquent donc qu'il est possible de bloquer le développement de la réponse inflammatoire et de protéger du choc septique à bactéries Gram-négatives en utilisant des thérapies ciblant TLR4. Par ailleurs, ils suggèrent qu'une fenêtre d'opportunité de plusieurs heures pourrait être mise à profit pour initier un traitement chez les patients septiques. Ces résultats devraient encourager la poursuite des essais cliniques en cours qui visent à tester l'efficacité de thérapies dirigées contre TLR4 comme traitement complémentaire de la sepsis.

La sala de cine vacía y el espectador expulsado

El presente trabajo aborda el problema que plantea el plano fijo de cinco minutos de una sala de cine vacía, que aparece hacia el final de la película Goodbye Dragon-Inn. Para entender las características del plano, en el contexto del film, se propone un recorrido por las distintas maneras en que el cine ha mostrado el interior de una sala de cine en la ficción, se establece una tipología de pantallas dentro de la pantalla y se reflexiona sobre cómo éstas han reflejado la figura del espectador. De esta manera, y en paralelo al estudio del mencionado plano, se van generando una serie de interrogantes (algunos resueltos y otros no) en torno a la situación del cine hoy en relación al espacio de proyección y su conexión con el espectador.

Studies on integrating kinematic design method with mechanical systems simulation techniques

Theultimate goal of any research in the mechanism/kinematic/design area may be called predictive design, ie the optimisation of mechanism proportions in the design stage without requiring extensive life and wear testing. This is an ambitious goal and can be realised through development and refinement of numerical (computational) technology in order to facilitate the design analysis and optimisation of complex mechanisms, mechanical components and systems. As a part of the systematic design methodology this thesis concentrates on kinematic synthesis (kinematic design and analysis) methods in the mechanism synthesis process. The main task of kinematic design is to find all possible solutions in the form of structural parameters to accomplish the desired requirements of motion. Main formulations of kinematic design can be broadly divided to exact synthesis and approximate synthesis formulations. The exact synthesis formulation is based in solving n linear or nonlinear equations in n variables and the solutions for the problem areget by adopting closed form classical or modern algebraic solution methods or using numerical solution methods based on the polynomial continuation or homotopy. The approximate synthesis formulations is based on minimising the approximation error by direct optimisation The main drawbacks of exact synthesis formulationare: (ia) limitations of number of design specifications and (iia) failure in handling design constraints- especially inequality constraints. The main drawbacks of approximate synthesis formulations are: (ib) it is difficult to choose a proper initial linkage and (iib) it is hard to find more than one solution. Recentformulations in solving the approximate synthesis problem adopts polynomial continuation providing several solutions, but it can not handle inequality const-raints. Based on the practical design needs the mixed exact-approximate position synthesis with two exact and an unlimited number of approximate positions has also been developed. The solutions space is presented as a ground pivot map but thepole between the exact positions cannot be selected as a ground pivot. In this thesis the exact synthesis problem of planar mechanism is solved by generating all possible solutions for the optimisation process ¿ including solutions in positive dimensional solution sets - within inequality constraints of structural parameters. Through the literature research it is first shown that the algebraic and numerical solution methods ¿ used in the research area of computational kinematics ¿ are capable of solving non-parametric algebraic systems of n equations inn variables and cannot handle the singularities associated with positive-dimensional solution sets. In this thesis the problem of positive-dimensional solutionsets is solved adopting the main principles from mathematical research area of algebraic geometry in solving parametric ( in the mathematical sense that all parameter values are considered ¿ including the degenerate cases ¿ for which the system is solvable ) algebraic systems of n equations and at least n+1 variables.Adopting the developed solution method in solving the dyadic equations in direct polynomial form in two- to three-precision-points it has been algebraically proved and numerically demonstrated that the map of the ground pivots is ambiguousand that the singularities associated with positive-dimensional solution sets can be solved. The positive-dimensional solution sets associated with the poles might contain physically meaningful solutions in the form of optimal defectfree mechanisms. Traditionally the mechanism optimisation of hydraulically driven boommechanisms is done at early state of the design process. This will result in optimal component design rather than optimal system level design. Modern mechanismoptimisation at system level demands integration of kinematic design methods with mechanical system simulation techniques. In this thesis a new kinematic design method for hydraulically driven boom mechanism is developed and integrated in mechanical system simulation techniques. The developed kinematic design method is based on the combinations of two-precision-point formulation and on optimisation ( with mathematical programming techniques or adopting optimisation methods based on probability and statistics ) of substructures using calculated criteria from the system level response of multidegree-of-freedom mechanisms. Eg. by adopting the mixed exact-approximate position synthesis in direct optimisation (using mathematical programming techniques) with two exact positions and an unlimitednumber of approximate positions the drawbacks of (ia)-(iib) has been cancelled.The design principles of the developed method are based on the design-tree -approach of the mechanical systems and the design method ¿ in principle ¿ is capable of capturing the interrelationship between kinematic and dynamic synthesis simultaneously when the developed kinematic design method is integrated with the mechanical system simulation techniques.

Optimising approaches to active immunotherapy of cancer

The design of therapeutic cancer vaccines is aimed at inducing high numbers and potent T cells that are able to target and eradicate malignant cells. This calls for close collaboration between cells of the innate immune system, in particular dendritic cells (DCs), and cells of the adaptive immune system, notably CD4+ helper T cells and CD8+ cytotoxic T cells. Therapeutic vaccines are aided by adjuvants, which can be, for example, Toll¬like Receptor agonists or agents promoting the cytosolic delivery of antigens, among others. Vaccination with long synthetic peptides (LSPs) is a promising strategy, as the requirement for their intracellular processing will mainly target LSPs to professional antigen presenting cells (APCs), hence avoiding the immune tolerance elicited by the presentation of antigens by non-professional APCs. The unique property of antigen cross-processing and cross-presentation activity by DCs plays an important role in eliciting antitumour immunity given that antigens from engulfed dead tumour cells require this distinct biological process to be processed and presented to CD8+T cells in the context of MHC class I molecules. DCs expressing the XCR1 chemokine receptor are characterised by their superior capability of antigen cross- presentation and priming of highly cytotoxic T lymphocyte (CTL) responses. Recently, XCR1 was found to be also expressed in tissue-residents DCs in humans, with a simitar transcriptional profile to that of cross- presenting murine DCs. This shed light into the value of harnessing this subtype of XCR1+ cross-presenting DCs for therapeutic vaccination of cancer. In this study, we explored ways of adjuvanting and optimising LSP therapeutic vaccinations by the use, in Part I, of the XCLl chemokine that selectively binds to the XCR1 receptor, as a mean to target antigen to the cross-presenting XCR1+ DCs; and in Part II, by the inclusion of Q.S21 in the LSP vaccine formulation, a saponin with adjuvant activity, as well as the ability to promote cytosolic delivery of LSP antigens due to its intrinsic cell membrane insertion activity. In Part I, we designed and produced XCLl-(OVA LSP)-Fc fusion proteins, and showed that their binding to XCR1+ DCs mediate their chemoattraction. In addition, therapeutic vaccinations adjuvanted with XCLl-(OVA LSP)-Fc fusion proteins significantly enhanced the OVA-specific CD8+ T cell response, and led to complete tumour regression in the EL4-OVA model, and significant control of tumour growth in the B16.0VA tumour model. With the aim to optimise the co-delivery of LSP antigen and XCLl to skin-draining lymph nodes we also tested immunisations using nanoparticle (NP)-conjugated OVA LSP in the presence or absence of XCLl chemokine. The NP-mediated delivery of LSP potentiated the CTL response seen in the blood of vaccinated mice, and NP-OVA LSP vaccine in the presence of XCLl led to higher blood frequencies of OVA-specific memory-precursor effector cells. Nevertheless, in these settings, the addition XCLl to NP-OVA LSP vaccine formulation did not increase its antitumour therapeutic effect. In the Part II, we assessed in HLA-A2/DR1 mice the immunogenicity of the Melan-AA27L LSP or the Melan-A26. 35 AA27l short synthetic peptide (SSP) used in conjunction with the saponin adjuvant QS21, aiming to identify a potent adjuvant formulation that elicits a quantitatively and qualitatively strong immune response to tumour antigens. We showed a high CTL immune response elicited by the use of Melan-A LSP or SSP with QS21, which both exerted similar killing capacity upon in vivo transfer of target cells expressing the Melan-A peptide in the context of HLA-A2 molecules. However, the response generated by the LSP immunisation comprised higher percentages of CD8+T cells of the central memory phenotype (CD44hl CD62L+ and CCR7+ CD62L+) than those of SSP immunisation, and most importantly, the strong LSP+QS21 response was strictly CD4+T cell-dependent, as shown upon CD4 T cell depletion. Altogether, these results suggest that both XCLl and QS21 may enhance the ability of LSP to prime CD8 specific T cell responses, and promote a long-term memory response. Therefore, these observations may have important implications for the design of protein or LSP-based cancer vaccines for specific immunotherapy of cancer -- Les vacans thérapeutiques contre le cancer visent à induire une forte et durable réponse immunitaire contre des cellules cancéreuses résiduelles. Cette réponse requiert la collaboration entre le système immunitaire inné, en particulier les cellules dendrites (DCs), et le système immunitaire adaptatif, en l'occurrence les lymphocytes TCD4 hdper et CD8 cytotoxiques. La mise au point d'adjuvants et de molécules mimant un agent pathogène tels les ligands TLRs ou d'autres agents facilitant l'internalisation d'antigènes, est essentielle pour casser la tolérance du système immunitaire contre les cellules cancéreuses afin de générer une réponse effectrice et mémoire contre la tumeur. L'utilisation de longs peptides synthétiques (LSPs) est une approche prometteuse du fait que leur présentation en tant qu'antigénes requiert leur internalisation et leur transformation par les cellules dendrites (DCs, qui sont les mieux à même d'éviter la tolérance immunitaire. Récemment une sous-population de DCs exprimant le récepteur XCR1 a été décrite comme ayant une capacité supérieure dans la cross-présentation d'antigènes, d'où un intérêt à développer des vaccins ciblant les DCs exprimant le XCR1. Durant ma thèse de doctorat, j'ai exploré différentes approches pour optimiser les vaccins avec LSPs. La première partie visait à cibler les XCR1-DCs à l'aide de la chemokine XCL1 spécifique du récepteur XCR1, soit sou s la forme de protéine de fusion XCL1-OVA LSP-Fc, soit associée à des nanoparticules. La deuxième partie a consisté à tester l'association des LSPs avec I adjuvant QS21 dérivant d'une saponine dans le but d'optimiser l'internalisation cytosolique des longs peptides. Les protéines de fusion XCLl-OVA-Fc développées dans la première partie de mon travail, ont démontré leur capacité de liaison spécifique sur les XCRl-DCs associée à leur capacité de chemo-attractio. Lorsque inclues dans une mmunisation de souris porteuse de tumeurs établies, ces protéines de fusion XCL1-0VA LSP-Fc et XCLl-Fc plus OVA LSP ont induites une forte réponse CDS OVA spécifique permettant la complète régression des tumeurs de modèle EL4- 0VA et un retard de croissance significatif de tumeurs de type B16-0VA. Dans le but d'optimiser le drainage des LSPs vers es noyaux lymphatiques, nous avons également testé les LSPs fixés de manière covalente à des nanoparticules co- injectees ou non avec la chemokine XCL1. Cette formulation a également permis une forte réponse CD8 accompagnée d'un effet thérapeutique significatif, mais l'addition de la chemokine XCL1 n'a pas ajouté d'effet anti-tumeur supplémentaire. Dans la deuxième partie de ma thèse, j'ai comparé l'immunogénicité de l'antigène humain Melan A soit sous la forme d un LSP incluant un épitope CD4 et CD8 ou sous la forme d'un peptide ne contenant que l'épitope CD8 (SSP) Les peptides ont été formulés avec l'adjuvant QS21 et testés dans un modèle de souris transgéniques pour les MHC let II humains, respectivement le HLA-A2 et DR1. Les deux peptides LSP et SSP ont généré une forte réponse CD8 similaire assoc.ee a une capacité cytotoxique équivalente lors du transfert in vivo de cellules cibles présentant le peptide SSP' Cependant les souris immunisées avec le Melan A LSP présentaient un pourcentage plus élevé de CD8 ayant un Phénotype «centra, memory» (CD44h' CD62L+ and CCR7+ CD62L+) que les souris immunisées avec le SSP, même dix mois après I'immunisation. Par ailleurs, la réponse CD8 au Melan A LSP était strictement dépendante des lymphocytes CD4, contrairement à l'immunisation par le Melan A SSP qui n'était pas affectée. Dans l'ensemble ces résultats suggèrent que la chemokine XCL1 et l'adjuvant QS21 améliorent la réponse CD8 à un long peptide synthétique, favorisant ainsi le développement d'une réponse anti-tumeur mémoire durable. Ces observations pourraient être utiles au développement de nouveau vaccins thérapeutiques contre les tumeurs.

Mellom tradisjon og spel : didaktikk for tekstil folkekunst

Denne avhandlinga er resultatet av eit behov for å forske på og utvikle ein didaktikk for tekstilfaget ved Folkekunststudiet, Institutt for folkekultur, Høgskolen i Telemark, Noreg. Studiet med folkekunst som fagfelt er eit relativt ungt studium på høgskolen, som vart etablert i 1984. Problemstillinga i denne avhandlinga er korleis ein kan utvikle ein forskingsbasert didaktikk der dei grunnleggjande prinsippa som særmerkjer den tradisjonelle folkekunsten, vert tekne vare på. I arbeidet med avhandlinga har eg prøvd på å klårgjere problemstillinga ut frå ulike perspektiv. Forskingsarbeidet har fokus på kommunikasjon og arbeidsmåtar i ljos av ulike teoriar. Det er ei hermeneutisk tilnærming som er vald for den didaktiske forståinga. Det fyrste drøftingstemaet har søkjeljos på kommunikasjon og dialog ved vidareføring av tekstil folkekunst. Både estetiske, teoretiske, praktiske og sosiale aspekt er nedfelte i læreplanen for studiet og skal utgjere grunnlaget for kommunikasjon og arbeidsmåtar. I den skapande og kopierande prosessen er det utvikla språklege reiskapar for både den sosiale og den estetiske sida der den teoretiske og praktiske faktoren er integrert. Møte med døme på tekstile tradisjonar så vel som praktisk forming av tekstilar har ført til refleksjon og dialog som involverer kontemplasjon, korrespondens og imaginasjon. Det andre temaet som er drøfta, er vidareføring av tradisjonelt visuelt formspråk. Her er merksemda retta mot kva som har skjedd formalt med ei gruppe tradisjonelle formelement i tekstilar i den institusjonelle vidareføringa over eit lengre tidsperspektiv. Resultatet syner at mange tradisjonelle formelement er borte frå den institusjonelle produksjonen. Formelementa kan ha fått ei meir naturalistisk utforming, eller dei er overførte til andre tradisjonelle tekstilteknikkar enn dei som var utgangspunktet. Rombeforma i den institusjonelle produksjonen er utført i færre variasjonar og kombinasjonar enn i den tradisjonelle produksjonen. Konklusjonen på drøftingstemaet er at spelereglar og spatialitet i høve til den formale komponenten i utvalet med tradisjonelle tekstilar ikkje er vidareført i alle gruppene av institusjonelle produkt. Resultatet kan få innverknad og fylgjer for utforming av ein framtidig didaktikk for faget. Arbeidsmåte og erfaring frå vidareføring av tekstile tradisjonar er det tredje temaet som er drøfta i avhandlinga. Kopiering og skapande prosessar er arbeidsmåtar som er brukte ved studiet i dag, og dei utgjer grunnlag for drøftingar i relasjon til vidareføring og erfaring. Konklusjonen er at i den skapande prosessen korrigerer tradisjonen utforminga, medan i kopiprosessen er eigen stilvilje og improvisering resultat av prosessane. Personar som deltek i prosessane, har sett seg sjølve og si historiske forankring inn i spelet, der visuelt og verbalt språk er resultat av integrasjon av tradisjonar. Dei tre drøftingstemaa utgjer grunnlaget for at didaktikk for folkekunst og tekstil kan bli intersubjektiv forståing av kva ein didaktikk for vidareføring av folkekunst med vekt på tekstil kan vere i utdanninga i dag. Indre og ytre dialog i skapande og kopierande prosessar femner om estetiske, materielle og tekniske faktorar sett i høve til spelereglar, spelerom, spatialitet og samspel i møte med tekstile tradisjonar. Samla utgjer det ein forskingsbasert didaktikk for faget der den overordna intensjonen er samtykke mellom tradisjon og spel.

Eleven som aktør i dialog med kunst : ungdoms erfaring med kunstundervisningens innhold og metode i faget kunst og håndverk i norsk grunnskole

Avhandlingen har sitt utspring i mitt engasjement for elevers møte med kunst i grunnskolen i faget kunst og håndverk og mitt syn på ungdom som kompetente bidragsytere til forskningen om fenomener som angår deres liv. Elevene er informanter til, eller aktører i, forskning på fenomenet dialog med kunst. Dialog med kunst er her definert som en helhetlig prosess som innlemmer alt fra elevenes møte med visuelle kunstverk til deres eget skapende arbeid. At avhandlingens fagdidaktiske problemområde er elevers praktisk skapende virksomhet, knytter undersøkelsen til slöjdpedagogisk forskning. Avhandlingens overgripende hensikt er å bidra til utvikling av fagdidaktikken i kunst og håndverk med utgangspunkt i elevenes erfaringer med kunstundervisningens innhold og metode på ungdomsskoletrinnet. Studien består av kasusstudier på to ungdomsskoler. Data ble innsamlet igjennom intervjuer, deltakende observasjon, dokumenter, prosessbøker og foto av formingsprodukter. Ungdoms dialog med kunst i skolen blir analysert og fremstilt ut fra et erfart og et operasjonalisert perspektiv. Funnene speiles i ulike fagdidaktiske tendenser, det vil si ulike hovedoppfatninger i debatten om det moderne samfunn, og i et virksomhetsteoretisk perspektiv. Resultatene fra undersøkelsen utfordrer oss til en fagdidaktisk nyorientering når det gjelder ungdoms møte med kunstverk i skolen, i retning av et mer ungdomskulturelt innhold og relasjonelle kunstmøter som er narrative, tolkningsorientert, opplevelsesorientert, dialogiske og flerstemmige. Undersøkelsen viser at elevene liker det praktisk skapende arbeidet, men at undervisningen i sterkere grad bør ta i bruk digital kunnskap og handle om hvordan kunst kan brukes som utgangspunkt for skapende arbeid, og den bør legge til rette for det læringspotensialet som ligger i dialogen elevene imellom. Elevene liker en undervisning som ikke bare handler om estetiske virkemidler, materialer og teknikker, men også om kommunikasjon og ytringsfrihet. Resultatene viser at det frie skapende arbeid består av tre likeverdige aspekter: det individuelle, det kulturelle og det sosiale. Både funnene og avhandlingens virksomhetsteoretiske perspektiv kan bidra til diskursen om kreativitetsbegrepet og identitetskonstruksjon i vårt moderne samfunn. Virksomhetssystemet blir i denne avhandlingen utviklet til en teori for skapende arbeid i faget kunst og håndverk, et overgripende fagdidaktisk rammeverk for bild/bildkonst og slöjdfaget satt inn i et nordisk utdanningsperspektiv.

Spectroscopic Studies of III-V Semiconductor Materials for Improved Devices

Defects in semiconductor crystals and at their interfaces usually impair the properties and the performance of devices. These defects include, for example, vacancies (i.e., missing crystal atoms), interstitials (i.e., extra atoms between the host crystal sites), and impurities such as oxygen atoms. The defects can decrease (i) the rate of the radiative electron transition from the conduction band to the valence band, (ii) the amount of charge carriers, and (iii) the mobility of the electrons in the conduction band. It is a common situation that the presence of crystal defects can be readily concluded as a decrease in the luminescence intensity or in the current flow for example. However, the identification of the harmful defects is not straightforward at all because it is challenging to characterize local defects with atomic resolution and identification. Such atomic-scale knowledge is however essential to find methods for reducing the amount of defects in energy-efficient semiconductor devices. The defects formed in thin interface layers of semiconductors are particularly difficult to characterize due to their buried and amorphous structures. Characterization methods which are sensitive to defects often require well-defined samples with long range order. Photoelectron spectroscopy (PES) combined with photoluminescence (PL) or electrical measurements is a potential approach to elucidate the structure and defects of the interface. It is essential to combine the PES with complementary measurements of similar samples to relate the PES changes to changes in the interface defect density. Understanding of the nature of defects related to III-V materials is relevant to developing for example field-effect transistors which include a III-V channel, but research is still far from complete. In this thesis, PES measurements are utilized in studies of various III-V compound semiconductor materials. PES is combined with photoluminescence measurements to study the SiO2/GaAs, SiNx/GaAs and BaO/GaAs interfaces. Also the formation of novel materials InN and photoluminescent GaAs nanoparticles are studied. Finally, the formation of Ga interstitial defects in GaAsN is elucidated by combining calculational results with PES measurements.

Out of the darkness and into the light : a study of inauthentic and authentic speech

Introduction Man can be described as the being who shows himself in speech, and from birth to death is continually speaking. Communication is so close to us, so woven into our very being, that we have little understanding of the way it is constituted; for it is as hard to obtain distance from communication as it is to obtain distance from ourselves. All communication is not alike. There are two basic modesl of communication, the inauthentic and the authentic, between which there occurs a constant tension. It is in the inauthentic mode, points out Heidegger, that we find ourselves "proximately and for the most part"; 1. Being and Time, pg. 68 Dasein decides as to the way it will comport itself in taking up its task of having being as an issue for it. " •.• it~, in its very being 'choose' itself and win itself; it can also lose itself and never win itself or only "seem" to do so. But only in so far as it is essentially something which can be authentic--that is, something of its own--can it have lost itself and not yet won itself." 2. therefore Heidegger also terms it "everydayness".2 Caught up in the world of everydayness, our speaking covers over and conceals3 our rootedness in being, leaving us in the darkness of untruth. The image of darkness may be inferred from Heidegger's use of the image of "clearing,,4 to depict being as 2. ibid. pg. 69 "Dasein's average everydayness, however, is not to be taken as a mere 'aspect'. Here too, and even in the mode of inauthenticity, the structure of existentiality lies ~ priori and here too Dasein's being is an issue for it in a definite way; and Dasein comports itself towards it the mode of average everydayness, even if this is only the mode of fleeing in the face of it and forgetfulness thereof." 3. ibid. pg. 59 "covering over" and "concealing" are 1;yays Dasein tries to flee its task of having being as an issue for itself. " ••• This being can be covered up so extensively that it becomes forgotten and no question arises about it or its meaning ••• n How everyday speaking accomplishes this will be taken up in detail in the second chapter which explores Dasein's everyday speech. 4. ibid, pg. 171 lI ••• we have in mind nothing other than the Existential - ontological structure of this entity (Dasein), that it is in such a way as to be its 'there'. To say that it is -' illuminated' [tlerleuchtet"] means that as Being-in-theworld it is cleared [gelichtetJ in itself7 not through any other entity, but in such a way that it is itself the clearing. Only for an entity which is eXistentially cleared in this way does what is present-at-hand become accessible in the light or hidden in the dark •••• " 3 dis-coveredness and truth. Our first task will be to explore the nature of communication in general and then to explore each of the modes manifested in turn. The structure of the inauthentic mode of communication can be explored by asking the following questions: What is this speaking about? Who is it that is speaking and who is spoken to? Does this speaking show man in his speech? The authentic mode is distinguished by the rarity with which we encounter it; as the inauthentic conceals, so the authentic reveals our rootedness in being. Yet this rarity makes it difficult to delineate its elusive structure clearly. Its constituent elements can be brought into focus by asking the same questions of this mode that we previously asked of the inauthentic mode. Our initial response to the disclosure of the authentic mode is to attempt to abandon the inauthentic mode and leave the darkness behind dwelling only in the "lighted place". All through the ages, some men pushing this to extreme, have, upon uncovering their relatedness to being, experienced a deep longing to dwell in such a "place" of pure truth and oft times denigrated or attempted to exclude the everyday world. Such 4. flight is twice mistaken: first it atbempts to fix truth as unchanging and static and secondly, it opposes this to untruth which it seeks to abolish. This is both the wrong view of truth and the wrong view of untruth as Heidegger points out in The Origin of The-Work of Art: The Way-to-be of truth, i.e., of discoveredness, is under the sway of refusal. But this refusal is no lack or privation, as if truth could be simply discoveredness rid of all covers. If it could be that, it would no longer be itself . ••• Truth in its way-to-be is untruth.5 Pure light is not the nature of Being nor is pure unconcealedness possible for man. Failure to remember this is the failure to realize that communication destroys itself in such flight because it no longer maintains the contingency of its task, i.e., the dis-closedness of being. We are reminded of the strong attraction this flight from darkness held for Plato. Light, truth and Being are all beyond the darkness and have nothing to do with it. In Book VII of the R~public, Socrates' explanation of the Allegory of the Cave to Glaucon points to a decided preference men have for the "lighted place". 5. The Origin Of The Work Of Art, pg. 42 5. Come then, I said, and join me in this further thought, and do not be surprised that those who attained to this height are not willing to occupy themselves with the affairs of men, but their souls ever feel the upward urge and yearning for that sojourn above. For this, I take it, is likely if in this point too the likeliness of our image holds. 6 Despite the attraction to pure truth, human communication is more complex than putting down one mode of communication and picking up another. Due to the fact that we are always on the way, the title of my thesis will have to be amended: OUT OF THE DARKNESS AND INTO THE LIGHT--AGAIN AND AGAIN. It must be this way because this is what it means to be human. This is the point made by Mephisto to Faust in pointing out that man, standing between God and the devil, needs both darkness and light: Er findet sich in einem ewigen Gl~t Uns hat er in die Finsternis gebracht, Und euch taugt einzig Tag und Nacht. 7 6. Republic z (517 c & d) It should be noted however, that while the philosopherking must be compelled to return to the cave for purely political reasons, once he has taken adequate view of the "brightest region of being" he has the full truth and his return to darkness adds nothing to the truth. 7. Faust, pg. 188 6. This thesis proposes to examine the grounds that give rise to communication, uncovering the structure of its inauthentic and authentic modes and paying close attention to tpeir interrelationship and to their relationship to language as "the house of Being": language that both covers and opens up man's rootedness in Being, transforming him as he moves along his way, taking up his "ownmost task" of becoming who he is. roots. He is the being who shows himself inn that reflects his forgetfulness or remembrance of his rootedness in being. Man comes into an already existent world and is addressedl through things in the world which are c

Niagara Falls Next to the River - Postcards

Board with six postcards of Niagara Falls. First postcard, In Victoria Park, Niagara Falls, Ont, Canada. Second postcard, entrance to Ontario Power Co. 's Works, Niagara Falls, Canada. Third postcard, The Refectory, Queen Victoria Park, Niagara Falls, Canada. Fourth postcard, Niagara Glen Inn, Niagara Glen, Canada. Fifth postcard, The Restaurant, Victoria Park, Niagara Falls, Canadian Side. Sixth postcard, The Administration Building, Victoria Park, Niagara Falls, Canada.

Photograph - Lorenzo Lamas, Laura Dickson, Donald Ziraldo

A photograph of Lorenzo Lamas, Laura Dickson and Donald Ziraldo at the Winetasting Inn on the Park November 1986.

Letter - E.C. Schmon to Arthur A. Schmon, 13 July 1917

The letter describes a party with family at the Pell Tree Inn where they had dinner and dancing. She also mentions that Arthur was made an instructor of topography and may soon do his examinations for second lieutenant.

Letter - E.C. Schmon to Arthur A. Schmon, 15 December 1918

The letter begins by describing her Red Cross uniform to be worn during the Xmas campaign. She also mentions an outing to see a show "Glorianna", then dancing at Murrays, then Hunter Island Inn for dinner. This letter is labelled number 202.