As sensing technology develops, it plays an important role in context-aware systems. Using context information improves the user experience of ubiquitous computing. One use of sensed information is to detect a meeting in progress in an office or a conference room. In our system, sensors gather context information from an office environment and act as a presence user agent to update a presence server with context changes. These context changes can be utilized by context-aware services. The presence messaging uses the SIP for Instant Messaging and Presence Leveraging Extensions (SIMPLE) protocol and the presence information is described in eXtensible Makeup Language (XML) format. In this thesis we present a context-sensing component that recognizes meetings in a typical office environment. A context-aware system is able to use this occupancy information to infer that the room is empty, an individual is alone in the room, or a meeting is taking place in the meeting room. Context-aware services might utilize this environmental information to automatically forward a user's incoming calls to their voice mail server. This and other example applications were developed to show the usefulness of this context information. / Så som sensor tekniken utvecklas, spelar de en viktig roll i kontextmedvetna system. Genom att använda kontextuell information förbättras användarupplevelsen av 'ubiquitous computing'. Ett användningsområde för sensorinsamlad information är att upptäcka ett möte som pågår i ett kontor eller konferenslokal. I vårt system samlar sensorer information från en kontorsmiljö och uppdaterar en närvaroserver med kontextuella förändringar. Dessa förändringar kan sedan utnyttjas av kontextmedvetna tjänster. För att förmedla den närvarostatusen använder närvaroservern SIP och ’Presence Leveraging Extensions’ (SIMPLE) protokoll. Närvaro information levereras i 'eXtensible Makeup Language' (XML) format. I denna avhandling presenterar vi en kontextsensorkomponent som känner av möten i en typisk kontorsmiljö. Ett kontextmedvetet system kan använda denna komponent för att dra slutsatsen att lokalen är tom, en person är ensam i lokalen, eller ett möte äger rum i lokalen. Kontextmedvetna tjänster kan utnyttja denna information för att automatiskt vidarebefordra en användares inkommande samtal till deras röstbrevlåda. Detta och andra exempel, har utvecklats för att visa nyttan av denna kontextuella information.

Le cancer de la vessie représente un vrai problème de santé publique, avec une surveillance et suivi clinique à long terme en raison de l’importance des fréquences de récidives. La chimiothérapie reste souvent inefficace. L’objectif de cette thèse est donc la recherche de marqueurs sérologiques et moléculaires à valeur pronostique dans le cancer de la vessie qui peuvent servir à prédire la maladie. D’abord, nous avons étudié trois facteurs de transcriptions des lymphocytes T activées qui sont T-bet, GATA-3 et Bcl-6. Nous avons montré une surexpression de T-bet chez les malades à stade invasif et de haut grade, cependant, la surexpression de GATA-3 et Bcl-6 a été corrélée au stade superficiel et de bas grade. La survie a été corrélée avec le groupe des malades sans histoires de récidive ou progression et avec la surexpression de Bcl-6 et GATA-3. Cependant les malades qui expriment fortement T-bet répondent mieux au BCG. Ensuite, nous avons visé la détection de FasL et TRAIL solubles dans le sérum des malades atteints du cancer vésicale. Nous avons montré une surexpression de sFasL et sTRAIL chez les malades à stade superficiel et de bas grade. Le rôle anti-tumoral de ces cytokines a été confirmé sur deux lignées du cancer de la vessie montrant que le traitement avec le sérum riche en sFasL ou en sTRAIL diminue la viabilité cellulaire in vitro. A la fin de cette thèse, nous avons testé l’activation p-Akt dans la tumeur vésicale. Nous avons montré une surexpression de p-Akt au sein des tumeurs comparées au tissu sain adjacent, et au sein des malades à stade invasif et de haut grade. Akt semble être un marqueur de progression tumorale dans le cancer de la vessie. / Bladder cancer is the first most common urogenital cancer in men in Tunisia, with a high recurrence rate. Patients with muscle-invasive disease develop metastasis. The need for expensive continuous surveillance. In this thesis we try to search some candidate biomarkers. Their use for cancer staging and personalization of therapy at the time of diagnosis in order to identify a better treatment could improve patient care. The aim of this first part of our study was to investigate the clinical significance of three immune cell-related transcription factors, T-bet, GATA-3 and Bcl-6 in Tunisian patients with bladder cancer. We found that T-bet level was significantly higher in invasive carcinoma with high- grade. However, T-bet is predictive of response to BCG. On the contrary, the expression of GATA-3 and Bcl-6 was significantly higher in non-invasive carcinoma with low grade. We furthermore studied the effect of activation of soluble FasL and TRAIL molecule in bladder cancer. We demonstrate that the mean serum level of sFasL was higher in patients than in normal donors. sFasL was only higher than in sera of healthy donors where patients had superficial stage and low- and medium-grade cancer. sTRAIL was significantly lower in sera from patients with invasive and high-grade bladder carcinoma than in controls. Finally, we demonstrate that p-Akt levels in patients with invasive carcinoma and high-grade bladder cancer were significantly elevated compared to patients with non-invasive and low grade bladder cancer. Altogether, our results suggest that Akt activation can provide useful prognostic information.

Made available in DSpace on 2016-04-28T20:37:35Z (GMT). No. of bitstreams: 1 Eduardo Goldenstein.pdf: 419720 bytes, checksum: f5e2cdb1bed51504885c6b81f7885255 (MD5) Previous issue date: 2011-05-10 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / As medicine has become increasingly scientific and technology-driven, medical care costs have increased and physicians had to specialize more and more, while working conditions have deteriorated for most of them. Because of the country s health policies, a vast majority of physicians took jobs in public hospitals where they do day and night duty shifts. Although salaries are low in the public sector, their tenure and labor rights are assured. In order to earn enough to meet their needs, a large part of these physicians work long hours, in day and night shifts, often times with no possibility of resting or engaging in leisure activities in their free time. This research focuses on physicians leisure. It was designed to understand how a group of physicians working in a public high-tech children s hospital in the city of São Paulo use their free time for leisure, and especially what their understanding of leisure is. Those physicians were asked to fill out a questionnaire and mark the number of working hours and the number of hours dedicated to other activities such as sleeping, resting, being with their families, and especially leisure considering two situations: a typical week and an atypical week. With this data in hand the author contacted seven physicians and long interviews were conducted to discuss the topic, i.e. leisure and physicians. Six of these seven physicians are pediatricians who passed an open competition and work duty shifts at the hospital s emergency room. One is a female plastic surgeon at the hospital where she also works as a volunteer. To analyze the interviews a specific methodology for qualitative research was used which is based on Heidegger s hermeneutic ontology. Based on the quantitative research the author concluded that although physicians believed they had leisure time this was not borne out by the facts. The data obtained in the qualitative research in turn allowed the author to conclude that for the group interviewed enjoying leisure time or not was directly related with the history or background of each one of them. For many of them leisure was yet another form of work, that is, it followed the same type of capitalistic logic, the logic of the perpetuum mobile of production. Leisure was also dependent on their availability to engage in a certain type of activity that could be considered as leisure or not. It also became apparent in the interviews that the physicians of this group had no difficulty in thinking about their leisure, although they were surprised with the topic at first. One last conclusion was that for this group of physicians leisure was not always equated with quality of life, or with pleasure or relaxation / Na medida em que a medicina atual foi se tornando mais e mais científica e tecnológica, seus custos foram se elevando, os médicos obrigados a cada vez mais se especializarem e as condições de trabalho para a grande maioria destes médicos se deteriorando. Por conta das políticas de saúde vigentes a maioria dos médicos do país passaram a trabalhar em hospitais ligados a rede pública em regime de plantões diurnos e noturnos, já que nesse setor, apesar de baixos salários, eles têm garantido estabilidade de emprego e direitos trabalhistas. Para poder fazer frente às suas necessidades financeiras, uma grande parte desses médicos trabalham muitas horas, diurnas e noturnas, muitas vezes sem possibilidade de descansar e ocupar seu tempo livre com atividades de lazer. O foco dessa pesquisa é o lazer dos médicos. O interesse é poder entender a forma como um grupo de médicos lotados em um hospital público infantil de alta tecnologia da cidade de São Paulo disponibilizam seu tempo livre para o lazer e, especialmente, a compreensão que eles têm desse lazer. Para tanto, inicialmente solicitamos aos médicos lotados na instituição que preenchessem um questionário onde se pedia que assinalassem tanto o número de horas voltadas para o trabalho quanto o número de horas voltadas para outras atividades como dormir, descansar, estar com a família e especialmente ao lazer, considerando-se duas situações diferentes: uma semana atípica e uma semana atípica. Com esses dados em mãos foram contatados sete médicos com os quais foram feitas entrevistas de longa duração a respeito do tema proposto, o lazer entre os médicos. Para a análise destas entrevistas adotou-se uma metodologia específica de pesquisa qualitativa referenciadas pela ontologia hermenêutica de Heidegger. A partir da pesquisa quantitativa pudemos concluir que apesar dos médicos pesquisados considerarem dispor sempre de tempo para o lazer isto não se confirmava na prática. Já os dados obtidos com a pesquisa qualitativa nos permitiram concluir que, para este grupo de médicos entrevistados a vivência ou não do lazer estava diretamente relacionada à história e experiência de vida de cada um deles; que para muitos o lazer se constituía numa outra forma de trabalho, ou seja, seguia a mesma lógica capitalista do moto contínuo de produção; e ainda que o lazer dependia da disponibilidade de cada um para um certo tipo de atividade que poderia ou não ser considerado lazer. Também ficou patente com a entrevista que os médicos desse grupo não tinham, apesar da surpresa inicial para com o tema, qualquer dificuldade de pensar no seu lazer. Por último, foi possível concluir que para este grupo de médicos nem sempre o lazer significava uma boa qualidade de vida, nem sempre se confundia com prazer ou relaxamento

Made available in DSpace on 2016-04-28T20:39:56Z (GMT). No. of bitstreams: 1 Peggy Coelho Carvalho.pdf: 501518 bytes, checksum: 6149c62f1458e0b29d66c0c53c49c3ac (MD5) Previous issue date: 2008-12-19 / This work aims to investigate some functions of silence on the psychoanalytic pratice. In his psychoanalytic space, where the core of the work is talking, dialoguing, the silent experiences can upset the analyst s capacity of observation and apprehension of the psychic reality, since some observable elements become unavailable. The adopted method is my own poetic style and, through the poetic bond established with the reader, I propose to expand the knowledge, reflections and comprehension of silent situations. I make an analogy with the musical technique of counterpoint using trhee principal and simultaneous events, or voices: Beethoven s, John Milton s and my own, interrelated around the theme of silence during all the narrative. My hypothesis is that it is possible that the being/not being can reveal and/or hide itself through silence, pregnant of mental pain. In this trajectory, I did not come across the silence by itself, but with the process of silence and, at the end, I found the silence as mental pain and pleasure / O objetivo deste trabalho é investigar algumas funções do silenciar na clínica psicanalítica. Neste espaço psicanalítico, no qual o cerne do trabalho é o conversar, o dialogar, as experiências silenciosas podem dificultar a capacidade de observação e apreensão da realidade psíquica por parte do analista, uma vez que alguns elementos observáveis ficam indisponíveis. O método adotado é o meu próprio estilo poético e, através do vínculo poético estabelecido com o leitor, proponho expandir nossos conhecimentos, nossas reflexões e compreensão sobre as situações silenciosas. Faço analogia com a técnica contrapontística musical no sentido de que utilizo três principais acontecimentos simultâneos, ou, vozes: de Beethoven, de John Milton e a minha própria voz que se inter-relacionam em torno do tema do silenciar durante toda a narrativa. Minha hipótese é a de que seja possível que o ser/não ser possa revelar-se e/ou ocultar-se através do silêncio, pregnante de dor mental. Nesta trajetória, fui deparando não com o silêncio, coisa-em-si, mas com o processo de silenciar e, ao final, encontrei o silenciar como dor e prazer mentais

Made available in DSpace on 2016-12-23T13:56:09Z (GMT). No. of bitstreams: 1 Dissertacao Fernanda Magri de Carvalho PDF.pdf: 4813468 bytes, checksum: f874502e73ece86d6ada6693a89b3fc4 (MD5) Previous issue date: 2009-03-19 / A incidência do carcinoma hepatocelular (CHC) apresenta grandes variações geográficas associadas a diferentes prevalências dos seus fatores etiológicos mais comuns. Embora não se conheça a sua real incidência no Espírito Santo, o CHC é freqüentemente diagnosticado no estado, associado à infecção com os vírus da hepatite B (VHB) e da hepatite C. No entanto, cerca de 40% dos casos não apresentam evidências destes fatores etiológicos. Por essa razão, buscou-se verificar se outro fator aflatoxinas poderia estar envolvido na etiologia do CHC no ES. Nas regiões tropicais, as aflatoxinas são consideradas um importante fator etiológico do CHC, especialmente quando associadas à infecção com o VHB. O objetivo da presente investigação foi estudar a prevalência da mutação específica ser-249 no gene TP53, cuja presença é um indicador indireto de contaminação alimentar com estas micotoxinas e sua relação com a infecção pelo VHB. Foram investigadas 41 amostras de CHC e 74 amostras de cirrose hepática sem CHC, utilizando-se a metodologia de PCR-RFLP e sequenciamento. O DNA do VHB foi pesquisado com iniciadores específicos para seis diferentes regiões do seu genoma. Os iniciadores utilizados foram escolhidos considerando as regiões mais conservadas do genoma viral e englobando os oito genótipos já identificados do VHB. A presença do DNA do VHB foi observada em 46% (19/41) dos casos de carcinoma hepatocelular estudados. A mutação ser-249 TP53 foi observada em 15% (6/41) e 1,5% (1/69) dos casos de CHC e de cirrose hepática sem CHC associado, respectivamente, não havendo correlação estatisticamente significativa com a presença do DNA do VHB. Foram encontradas outras quatro mutações no códon 250 do gene TP53. A prevalência total de 24% (10/41) de mutações ser-249 e mutações no códon 250 nos pacientes com carcinoma hepatocelular possivelmente também associadas à contaminação com aflatoxinas pode indicar a provável exposição alimentar às aflatoxinas no Espírito Santo / The hepatocellular carcinoma (HCC) incidence has large geographic variation, which is associated with the different prevalences of its most common etiological factors. Although its real incidence in the Espirito Santo State is not known, the HCC is frequently diagnosed in ES associated with hepatitis B (HBV) and C virus infections. However, approximately 40% of the cases do not present known etiological factors. Therefore, it was the purpose of this study to verify if another factor - aflatoxins - could be involved in the HCC etiology in ES. In tropical areas, the aflatoxins are considered an important etiological factor of CHC, especially when associated with HBV infection. The aim of this investigation was to study the prevalence of the specific ser-249 mutation in the TP53 gene was studied, that indicates indirect food contamination with aflatoxins and its association with HBV infection. Forty-one HCC samples and seventy-four hepatic cirrhosis samples were investigated, using PCRRFLP methodology and sequencing. The HBV DNA was amplified with specific primers targeting six different regions of its genome. Primers were chosen considering the most conserved regions of the viral genome and encompassing the eight known genotypes of HBV. Presence of HBV DNA was observed in 46% (19/41) of the hepatocellular carcinoma cases studied. The ser-249 TP53 mutation was observed in 15% (6/41) and 1,5% (1/69) of the HCC and cirrhosis cases respectively, not having statistical correlation with the presence of HBV DNA. Other four mutations in TP53 codon 250 were found, yielding a total mutation prevalence of 24% (10/41), considering both ser-249 and codon 250 mutations, possibly also associated with food contamination by aflatoxins, and suggesting an alimentary exposition to aflatoxins in Espirito Sant

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2016-09-26T11:26:03Z No. of bitstreams: 2 Tese - Ema Marta Dunck Cintra - 2016.pdf: 9425370 bytes, checksum: 2c06326bfc683abfeeea650905ca39a0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-26T11:52:58Z (GMT) No. of bitstreams: 2 Tese - Ema Marta Dunck Cintra - 2016.pdf: 9425370 bytes, checksum: 2c06326bfc683abfeeea650905ca39a0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-26T11:52:58Z (GMT). No. of bitstreams: 2 Tese - Ema Marta Dunck Cintra - 2016.pdf: 9425370 bytes, checksum: 2c06326bfc683abfeeea650905ca39a0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-08-26 / How can an ethnic identity that has been wiped out, numbed, silenced by colonization process be identified? In the case of the Chiquitano in the Portal do Encantado-MT, who had positioned themselves as indigenous people to have the right to remain in their territory, the first strategy was to trigger the mother tongue which had taken refuge in the memory of their elders. Thus, the focus of this thesis is to present how, through “memory bilingualism” , this people could trigger an interactive process of resistance, cultural and political positioning of identity assertion. An identity that those who remembered (the elders) were able to reconstruct, given the common unitary point (belonging) of representation and impulsion to the experience of traditional knowledge. This knowledge, inherited from a common past, provoked/articulated/triggered a fight in view of the hegemony imposed by the settlers and their ways of homogenization of knowledge, culture and life. Hence, based on the results of this study, which entangles decolonial authors and authors who treat the language as discourse, it is possible to state that the Chiquitano, when triggered the mother tongue which was stored in their memory, could reach the dwelling for their being and gradually establish a decolonial process, identifying their identity and their place of belonging. That’s because, collectively, they acted and promoted the interaction of different generations of people. Through reflection, they found themselves, creating themselves and their “new” world. By demanding their own school, they produced science. Giving value to the ancestral knowledge, they defined the curriculum and pedagogic practices. In this way, they recorded their culture, which was dormant in the language memory. They produced an authorial and coping pedagogy. The valorization of their knowledge and their being created the possibilities for fighting the dominant power. Memory bilingualism was the guiding basis for the training of teachers and their pedagogic practices, as well as for the assertion of an identity. Everything together, provided the resurgence of a “being more” (FREIRE, 2014) and led the sociocultural vitalization of the Chiquitano do Brasil in the Portal de Encantado-MT. / Cómo marcar una identidad étnica apagada, dormida, silenciada a causa del proceso colonizador? En el caso de los Chiquitano del Portal do Encantado-MT, que necesitaron posicionarse como indígenas para tener el derecho de permanecer en su territorio, la primera estrategia fue accionar la lengua materna que estaba refugiada en la memoria de los ancianos. Es de eso que trata esta tesis, al presentar cómo, a través del “bilingüismo de memoria” ese pueblo consiguió desarrollar un proceso interactivo de resistencia, posicionamiento cultural y político de afirmación de la identidad. Esta, por su vez, se dio a través de los ancianos que recordaban la lengua dado el punto de unidad (pertenencia) común de representación y de impulsión para la vivencia de los saberes tradicionales. Esos saberes, heredados de un pasado común, provocaron/articularon/accionaron una lucha en contra de la hegemonía impuesta por el colonizador y sus formas de homogenización del conocimiento, de la cultura y de la vida, De esta manera, con base en los resultados de este estudio, que articula autores decoloniales y autores que tratan de la lengua como discurso, es posible afirmar que los Chiquitano, al accionar la lengua materna que estaba guardada en su memoria, lograron llegar a la morada de su ser y paulatinamente establecer un proceso decolonial, demarcando su identidad y su lugar de pertenencia. Eso porque, colectivamente, actuaron y promovieron la interacción de las diferentes generaciones del pueblo. Reflexionando, fueron reencontrándose, constituyéndose a sí mismos y a su “nuevo” mundo. Con la solicitud de una escuela propia para ellos, produjeron ciencia. Valorando el conocimiento ancestral, definieron el currículo y las prácticas pedagógicas. Registraron, así, su cultura, que estaba dormida en la memoria de la lengua. Produjeron una pedagogía autoral y de enfrentamiento. La valoración de su saber y de su ser creó las posibilidades de lucha frente al poder dominante. El bilingüismo de memoria fue la base orientadora para la formación de los profesores y de sus prácticas pedagógicas, así como para la formación de una identidad. Todo eso, en conjunto, propició el resurgimiento de un “ser más” (FREIRE, 2014) y provocó la vitalización sociocultural de los Chiquitano de Brasil en el Portal do Encantado- MT. / Como marcar uma identidade étnica apagada, adormecida, silenciada em virtude do processo colonizador? No caso dos Chiquitano do Portal do Encantado-MT, que tiveram de se posicionar como indígenas para terem o direito de permanecer em seu território, a primeira estratégia foi acionar a língua materna que estava refugiada na memória dos mais velhos. E é disso que esta tese trata, ao apresentar como, por meio do “bilinguismo de memória” , esse povo conseguiu desencadear um processo interativo de resistência, posicionamento cultural e político de afirmação da identidade. Identidade essa que os lembradores (anciãos) puderam reconstituir, dado o ponto unitário (pertencimento) comum de representação e de impulsão para a vivência dos saberes tradicionais. Esses saberes, herdados de um passado comum, provocaram/articularam/acionaram uma luta em face da hegemonia imposta pelo colonizador e suas formas de homogeneização do conhecimento, da cultura e da vida. Assim, com base nos resultados deste estudo, que enreda autores decoloniais e autores que tratam a língua como discurso, é possível afirmar que os Chiquitano, ao acionarem a língua materna que estava guardada em sua memória, puderam chegar à morada do seu ser e gradativamente estabelecer um processo decolonial, demarcando sua identidade e seu lugar de pertencimento. Isso porque, coletivamente, agiram e promoveram a interação das diferentes gerações do povo. Refletindo, foram se reencontrando, constituindo-se a si mesmos e o seu “novo” mundo. Ao demandarem uma escola própria para eles, produziram ciência. Valorizando o conhecimento ancestral, definiram o currículo e as práticas pedagógicas. Registraram, assim, a sua cultura, que estava adormecida na memória da língua. Produziram uma pedagogia autoral e de enfrentamento. A valorização do seu saber e do seu ser criou as possibilidades de luta diante do poder dominante. O bilinguismo de memória foi a base orientadora para a formação dos professores e para a fundamentação de suas práticas pedagógicas, assim como para a afirmação de uma identidade. Tudo isso, associado, propiciou o ressurgimento de um “ser mais” (FREIRE, 2014) e provocou a vitalização sociocultural dos Chiquitano do Brasil no Portal de Encantado-MT.

Made available in DSpace on 2017-07-10T18:26:26Z (GMT). No. of bitstreams: 1 Tiago Monteiro Violante.pdf: 313159 bytes, checksum: 7044975ab27c799a0e9e21c94af00aea (MD5) Previous issue date: 2011-08-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / One of the constants of Sartre's theoretical concerns is the question of the constitution of the ego. It can be seen a detailed siege by the French philosopher, in search of this existential problem par excellence: the relationship of consciousness to self. The ego is a structure of consciousness immanent constitute or is a product of a transcendent poetic creation held by such transcendental field? What this paper intends to present is how Sartre captures the features, functions and meanings that the ego is in his existentialism. It is necessary, however, cut this issue on the horizon of an argumentative progression on the foundations of human reality, presented since his first work, The Transcendence of the Ego, to his most important and significant, Being and Nothingness. / Uma das constantes preocupações teóricas de Sartre é a questão da constituição do ego. Percebe-se uma detalhada perseguição, por parte do filósofo francês, em busca desta problemática existencial por excelência: a relação da consciência com seu eu. O ego é uma estrutura imanente da consciência constituinte, ou é um produto transcendente de uma criação poética realizada por tal campo transcendental? O que este trabalho pretende apresentar é como Sartre apreende as características, funções e sentidos que o ego assume em seu existencialismo. Torna-se necessário, entretanto, recortar esta temática no horizonte de uma progressão argumentativa sobre os fundamentos da realidade humana, apresentada desde sua primeira obra, A transcendência do Ego, até sua obra mais importante e expressiva, O Ser e o Nada.

No description available.

Collagen is one of the most abundant human proteins. It exists as a right-handed superhelix called the triple helix. The triple helix consists of three left-handed polyproline type II (PPII helices) that intertwine around a common axis. Each PPII helix has the repeating peptide sequence (Gly–Xaa–Yaa)n with a high content of (2S)-proline (Pro) in the Xaa position (ca. 28%) and (2S,4R)-hydroxyproline (Hyp) in the Yaa position (ca. 38%). Unique to the prolyl amide is the ease of cis-trans isomerization. Since the triple helix necessitates that all peptide bonds be in the trans conformation, isomerization is the rate-limiting step in collagen folding. However, eliminating isomerization with a trans-locked alkene isostere destabilizes collagen-like peptides. Collagen is stabilized by electronic interactions, namely the n→π* interaction. Halo-alkene isosteres may be used to recapture these electronic interactions and stabilize a collagen-like peptide. An in-depth conformational analysis was conducted at the MP2/6-311+G(2d,p) level of theory to determine the viability of conformationally-locked halo-alkene isosteres. Fluoro-alkenes and chloro-alkenes were modeled at both the Gly–Pro and Pro–Pro (as a Pro–Hyp mimic) amide positions. Compared to the collagen crystal structure PDB ID: 1K6F, we found the fluoro-alkenes were closer geometric matches to both Gly–Pro and Pro–Pro than the corresponding chloro-alkenes. The chloro-alkene was predicted to have stronger n→π* interactions. The trans-locked proteo-alkene was also analyzed to understand why it destabilized the triple helix. We found that these models had other local minima close to the desired PPII geometry, likely leading to enhanced backbone flexibility. This deleterious flexibility was not predicted for either fluoro-alkene or chloro-alkene models. The conformationally-locked halo-alkene isostere Fmoc–Gly–Ψ[(Z)CF=C]-Pro–Hyp(tBu)–OH was designed and synthesized as a (Z)-fluoro-alkene Gly–Pro isostere. We used the chiral catalyst, L-Thr, for asymmetric aldol addition to cyclopentanone, which inadvertently enhanced the yield of the wrong enantiomer, in contrast with aldol addition to cyclohexanone. A Mg2+-promoted Horner-Wadsworth-Emmons reaction afforded the (Z)-fluoro-alkene over the (E)-fluoro-alkene in about a 2:1 ratio. The two diastereomers, Fmoc–Gly–Ψ[(Z)CF=C]-L-Pro–Hyp(tBu)–OH and Fmoc–Gly–Ψ[(Z)CF=C]-D-Pro–Hyp(tBu)–OH were separated by supercritical CO2 chromatography. The collagen-like peptides Ac–(Gly–Pro–Hyp)3–Gly–Ψ[(Z)CF=C]-L-Pro–Hyp–(Gly–Pro–Hyp)4–Gly–Gly–Tyr–NH2, Ac–(Gly–Pro–Hyp)3–Gly–Ψ[(Z)CF=C]-D-Pro–Hyp–(Gly–Pro–Hyp)4–Gly–Gly–Tyr–NH2, and the control peptide Ac–(Gly–Pro–Hyp)8–Gly–Gly–Tyr–NH2 were synthesized on solid-phase resin. The CD spectra of all three peptides showed the characteristic collagen triple-helix signature. The folding stability was determined by thermal melting (Tm). The peptide with the fluoro-alkene guest, Gly–Ψ[(Z)CF=C]-L-Pro–Hyp, was found to have a Tm value of 42.2 °C. The Tm of the control peptide was found to be 49.0 °C, a difference in stability of only ΔTm –6.8. Thus, the (Z)-fluoro-alkene as a Gly–Pro isostere forms a relatively stable triple helix. The peptide with the Gly–Ψ[(Z)CF=C]-D-Pro–Hyp guest was shown to have a linear relationship between ellipticity and temperature, indicating that a stable triple helix did not form. The enhanced stability of the (Z)-fluoro-alkene compared to the (E)-alkene Gly–Pro isostere (Tm = 28.3 °C) may be due to a stabilizing n→π* interaction, as determined by NMR deshielding of the 19F nucleus in the collagen-like peptide. In biological systems, isomerization of the prolyl amide is catalyzed by enzymes called PPIases. The PPIase Pin1 specifically catalyzes isomerization of the pSer–Pro sequence from the cis-conformation to the trans-conformation. Pin1 plays a crucial role in the G2→M transition of the cell cycle, implying the importance of cis-trans isomerization. The dipeptides H–Ser–Ψ[(Z)CH=C]-Pro–OH, H–Ser–Ψ[(E)CH=C]-Pro–OH and native H–Ser–Pro–OH were synthesized by literature methods, and activated for aminoacylation of tRNACUA for in vitro transcription-translation. Aminoacylation by chemical methods required the synthesis of a pdCpA dinucleotide. Formation of the dipeptide-dinucleotide complex was not completed because protection of the Ser side chain was problematic. On the other hand, conversion of the dipeptide into the 3,5-dinitrobenzyl ester conjugate allowed for enzymatic aminoacylation using the dFx flexizyme, an RNA enzyme. The native dipeptide was successfully coupled to tRNACUA and is ready for incorporation into a full-length Bora protein by in vitro transcription-translation. Both cis- and trans-locked alkene mimics have been converted to their respective 3,5-dinitrobenzyl ester conjugates. / Doctor of Philosophy / The proline amide (Xaa–Pro) in peptides and proteins is unique in that it allows for cis-trans isomerization. The triple-helix region of human collagen consists mostly of the repeating sequence (Gly–Pro–Hyp)n. Xaa–Pro amide-bond isomerization is rate-limiting for triple-helix formation. We eliminated isomerization at one position in a collagen-like peptide with a locked alkene mimic of Gly–Pro to attempt to stablize the triple-helix. Our computational results predicted that a fluoro-alkene Gly–Pro isostere would be a close geometric match for the native amide. Experimental results showed that a collagen-like peptide with a fluoro-alkene Gly–Pro isostere has an unfolding temperature that is 6.9 °C lower than the native control peptide. 19F NMR data of the collagen-like peptide shows a surprising deshielding of the fluorine nucleus, suggesting its participation in a stabilizing n→π* electronic interaction, similar to the native amide. Isomerization also plays a key role in proper cell division. We followed established methods to synthesize the cis- and trans-locked alkene mimics of Boc–Ser–Pro–OH and converted them into the 3,5-dinitrobenzyl ester conjugates. The 3,5-dinitrobenzyl ester is recognized by the dinitrobenzyl flexizyme (dFx) for enzymatic aminoacylation of tRNA. Once the alkene isosteres are aminoacylated, they will be incorporated into a full-length cell cycle regulatory protein called Bora to determine whether the cis- or trans-Pro state is necessary for healthy human mitosis, and which results in cancerous human mitosis.

L’augmentation de la densité et la réduction de la tension d’alimentation des circuits intégrés rend la contribution des effets singuliers induits par les radiations majoritaire dans la diminution de la fiabilité des composants électroniques aussi bien dans l’environnement radiatif spatial que terrestre. Cette étude porte sur la modélisation des mécanismes physiques qui conduisent à ces aléas logiques (en anglais "Soft Errors"). Ces modèles sont utilisés dans une plateforme de simulation,appelée TIARA (Tool suIte for rAdiation Reliability Assessment), qui a été développée dans le cadre de cette thèse. Cet outil est capable de prédire la sensibilité de nombreuses architectures de circuits (SRAM,Flip-Flop, etc.) dans différents environnements radiatifs et sous différentes conditions de test (alimentation, altitude, etc.) Cette plateforme a été amplement validée grâce à la comparaison avec des mesures expérimentales effectuées sur différents circuits de test fabriqués par STMicroelectronics. La plateforme TIARA a ensuite été utilisée pour la conception de circuits durcis aux radiations et a permis de participer à la compréhension des mécanismes des aléas logiques jusqu’au noeud technologique 20nm. / Aggressive integrated circuit density increase and power supply scaling have propelled Single Event Effects to the forefront of reliability concerns in ground-based and space-bound electronic systems. This study focuses on modeling of Single Event physical phenomena. To enable performing reliability assessment, a complete simulation platform named Tool suIte for rAdiation Reliability Assessment (TIARA) has been developed that allows performing sensitivity prediction of different digital circuits (SRAM, Flip-Flops, etc.) in different radiation environments and at different operating conditions (power supply voltage,altitude, etc.) TIARA has been extensively validated with experimental data for space and terrestrial radiation environments using different test vehicles manufactured by STMicroelectronics. Finally, the platform has been used during rad-hard digital circuits design and to provide insights into radiation-induced upset mechanisms down to CMOS 20nm technological node.