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This dissertation reports on the technology of cell-free protein synthesis (CFPS) including 1) stabilized lyophilized cell-free systems and 2) enhanced heterogeneous cell extracts. This work further considers applications of CFPS systems in 1) rapid vaccine development, 2) functional virus-based nanoparticles, 3) site-specific protein immobilization, and 4) expanding the language of biology using unnatural amino acids. CFPS technology is a versatile protein production platform that has many features unavailable in in vivo expression systems. The primary benefit cell-free systems provide is the direct access to the reaction environment, which is no longer hindered by the presence of a cell-wall. The “openness" of the system makes it a compelling candidate for many technologies. One limitation of CFPS is the necessity of freezing for long-term viable storage. We demonstrate that a lyophilized CFPS system is more stable against nonideal storage than traditional CFPS reagents. The Escherichia coli-based CFPS system in this work is limited by the biocatalytic machinery found natively in E. coli. To combat these limitations, exogenous biocatalysts can be expressed during fermentation of cells prepared into extract. We demonstrate that simple adjustments in the fermentation conditions can significantly increase the activity of the heterogeneous extract. Towards virus-based particles and vaccines, we demonstrate that the open nature of CFPS can be utilized for coexpression of virus proteins and self-assembly of virus particles. This technique allows for the rapid production of potential vaccines and novel functional virus-based nanoparticles. Unnatural amino acids expand the effective language of protein biology. Utilizing CFPS as an expression system, we demonstrated that the incorporation of a single specific unnatural amino acid allows for site-specific immobilization, thus stabilizing the protein against elevated temperatures and chemical denaturants. Current unnatural amino acid incorporation technologies are limited to one or few simultaneous incorporations and suffer from low efficiency. This work proposes a system that could potentially allow for upwards of 40 unnatural amino acids to be simultaneously incorporated, effectively tripling the protein code. These projects demonstrate the power and versatility of CFPS technologies while laying the foundation for promising technologies in the field of biotechnology.

Made available in DSpace on 2016-04-26T20:30:29Z (GMT). No. of bitstreams: 1 Livia Cristina Rocha.pdf: 815766 bytes, checksum: 82635d5039ae0bb942d92f31298e9467 (MD5) Previous issue date: 2010-06-16 / This study aims to analyze the many issues raised in relation to the reassignment of sex in their ways: neocolpovulvoplastia and neofaloplastia. Among them: surgical inefficiency and the possibility of repentance, the body naturally created; finding injury of serious nature, and especially the consequences brought by this surgery in the legal world. The reassignment of sex is only compatible with the figure of the transsexual, except for his help in other cases the behavioral states of sexuality (intersex, hermaphroditism, homosexuality, bisexuality and transvestism). The transsexual is defined as the person who experience an ongoing conflict between their physical sex and psychological, in other words, their mental gender identity does not match with the biological, which leads the transsexual vehemently deny their sexual organs, and in some cases the injured themselves or commit suicide. Thus, the reassignment of sex is shown as the only proper means to repair the suffering of transsexuals and is the only effective treatment. She is the extent of body-mind balance and essential to the preservation of life, health, health (physical and mental) and well-being of the patient. Once accomplished, brings with anatomical transformation for the transsexual, but how it reflects the legal world? Currently, the transsexual surgery is recognized the right to change the first name and sex in its civil register. Emphasized that it is not a unanimous position. At this point, the course includes the three main current jurisprudence, the first one in favor of changing the civil registry and its dual knowledge (first name and gender), the second most favorable only to registration under the justification of preserving rights of third parties and continuity of existing Public and, finally, one that begs the insertion of the term "transsexual" in the cartouche would register, so it would be acting under the aegis of the veracity of public record. Beyond these points, see that the only way to promote social inclusion of transsexuals will be with the completion of the process begun with the reassignment of sex, which is just a phase, complemented with the conclusion of the civil registry change that, because only thus can preserve and realize the much-trumpeted principle of human dignity / O presente trabalho tem como objetivo analisar as questões suscitadas em relação à cirurgia de redesignação de sexo em suas modalidades: neocolpovulvoplastia e neofaloplastia. Dentre elas: ineficácia cirúrgica; possibilidade de arrependimento; naturalidade do órgão criado; constatação de lesão corporal de natureza grave; e, especialmente, os reflexos trazidos pela realização dessa cirurgia no mundo jurídico. A cirurgia de redesignação de sexo apenas é compatível com a figura do transexual, excluindo de seu amparo os demais casos de estados comportamentais da sexualidade (intersexualismo; hermafroditismo; homossexualismo; bissexualismo e travestismo). O transexual é definido como aquela pessoa que vivencia um conflito permanente entre seus sexos físico e psíquico, em outras palavras, sua identidade sexual psíquica não corresponde com a biológica, o que leva o transexual recusar veemente seus órgãos sexuais, chegando em alguns casos a lesionar-se ou suicidar-se. Destarte, a cirurgia de redesignação de sexo se mostra como o único meio hábil para reparar o sofrimento do transexual, constituindo a única forma de tratamento efetivo. Ela é o alcance do equilíbrio corpo-mente e essencial à preservação da vida, da saúde, da integridade (física e psíquica) e do bem-estar do paciente. Uma vez realizada, traz relevante transformação anatômica para o transexual, mas como isso reflete no mundo jurídico? Atualmente, ao transexual operado é reconhecido o direito à alteração do prenome e do sexo em seu registro civil. Ressalta que não é uma posição unânime. Neste ponto, serão estudadas as três principais correntes jurisprudências, a primeira delas favorável à alteração do registro civil e seu duplo conhecimento (prenome e sexo); a segunda delas favorável apenas à averbação, sob a justificativa de preservar direito de terceiros e a continuidade do registro público; e, por último, aquela que suplica pela inserção da denominação transexual na cártula registrária, assim estaria agindo sob a égide da veracidade do registro público. Ultrapassados esses pontos, verá que o único modo de promover a inclusão social do transexual será com a completude do processo iniciado com a cirurgia de redesignação de sexo, que é apenas uma fase, sendo complementada com a efetivação da alteração do registro civil daquele, pois, somente assim poderá preservar e efetivar o tão proclamado princípio da dignidade humana

De nombreux signaux physiques incluant des signaux audio (musique, parole), médicaux (ECG, PCG), de mammifères marins ou d'ondes gravitationnelles peuvent être modélisés comme une superposition d'ondes modulées en amplitude et en fréquence (modes AM-FM), appelés signaux multicomposantes (SMCs). L'analyse temps-fréquence (TF) joue un rôle central pour la caractérisation de tels signaux et, dans ce cadre, diverses méthodes ont été développées au cours de la dernière décennie. Néanmoins, ces méthodes souffrent d'une limitation intrinsèque appelée le principe d'incertitude. Dans ce contexte, la méthode de réallocation (MR) a été développée visant à améliorer les représentations TF (RTFs) données respectivement par la transformée de Fourier à court terme (TFCT) et la transformée en ondelette continue (TOC), en les concentrant autour des lignes de crête correspondant aux fréquences instantanées. Malheureusement, elle ne permet pas de reconstruction des modes, contrairement à sa variante récente connue sous le nom de transformée synchrosqueezée (TSS). Toutefois, de nombreux problèmes associés à cette dernière restent encore à traiter tels que le traitement des fortes modulations en fréquence, la reconstruction des modes d'un SMC à partir de sa TFCT sous-échantillonnée or l'estimation des signatures TF de modes irréguliers et discontinus. Cette thèse traite principalement de tels problèmes afin de construire des nouvelles méthodes TF inversibles plus puissantes et précises pour l'analyse des SMCs.Cette thèse offre six nouvelles contributions précieuses. La première contribution introduit une extension de TSS d'ordre deux appliqué à la TOC ainsi qu'une discussion sur son analyse théorique et sa mise en œuvre pratique. La seconde contribution propose une généralisation des techniques de synchrosqueezing construites sur la TFCT, connue sous le nom de transformée synchrosqueezée d'ordre supérieur (FTSSn), qui permet de mieux traiter une large gamme de SMCs. La troisième contribution propose une nouvelle technique utilisant sur la transformée synchrosqueezée appliquée à la TFCT de second ordre (FTSS2) et une procédure de démodulation, appelée DTSS2, conduisant à une meilleure performance de la reconstruction des modes. La quatrième contribution est celle d'une nouvelle approche permettant la récupération des modes d'un SMC à partir de sa TFCT sous-échantillonnée. La cinquième contribution présente une technique améliorée, appelée calcul de représentation des contours adaptatifs (CRCA), utilisée pour une estimation efficace des signatures TF d'une plus grande classe de SMCs. La dernière contribution est celle d'une analyse conjointe entre l'CRCA et la factorisation matricielle non-négative (FMN) pour un débruitage performant des signaux phonocardiogrammes (PCG). / Many physical signals including audio (music, speech), medical data (ECG, PCG), marine mammals or gravitational-waves can be accurately modeled as a superposition of amplitude and frequency-modulated waves (AM-FM modes), called multicomponent signals (MCSs). Time-frequency (TF) analysis plays a central role in characterizing such signals and in that framework, numerous methods have been proposed over the last decade. However, these methods suffer from an intrinsic limitation known as the uncertainty principle. In this regard, reassignment method (RM) was developed with the purpose of sharpening TF representations (TFRs) given respectively by the short-time Fourier transform (STFT) or the continuous wavelet transform (CWT). Unfortunately, it did not allow for mode reconstruction, in opposition to its recent variant known as synchrosqueezing transforms (SST). Nevertheless, many critical problems associated with the latter still remain to be addressed such as the weak frequency modulation condition, the mode retrieval of an MCS from its downsampled STFT or the TF signature estimation of irregular and discontinuous signals. This dissertation mainly deals with such problems in order to provide more powerful and accurate invertible TF methods for analyzing MCSs.This dissertation gives six valuable contributions. The first one introduces a second-order extension of wavelet-based SST along with a discussion on its theoretical analysis and practical implementation. The second one puts forward a generalization of existing STFT-based synchrosqueezing techniques known as the high-order STFT-based SST (FSSTn) that enables to better handle a wide range of MCSs. The third one proposes a new technique established on the second-order STFT-based SST (FSST2) and demodulation procedure, called demodulation-FSST2-based technique (DSST2), enabling a better performance of mode reconstruction. The fourth contribution is that of a novel approach allowing for the retrieval of modes of an MCS from its downsampled STFT. The fifth one presents an improved method developed in the reassignment framework, called adaptive contour representation computation (ACRC), for an efficient estimation of TF signatures of a larger class of MCSs. The last contribution is that of a joint analysis of ACRC with non-negative matrix factorization (NMF) to enable an effective denoising of phonocardiogram (PCG) signals.

L'objectif de cette thèse est d'optimiser l'utilisation d'actionneurs redondants pour un avion de transport lorsqu’une défaillance des actionneurs arrive en vol. La tolérance aux pannes résulte ici de la redondance des actionneurs présents sur l’avion. Différents concepts et méthodes classiques liés aux chaînes de commande de vol tolérantes aux pannes sont d'abord examinés et de nouveaux concepts utiles pour l'analyse requise sont introduits. Le problème qui est abordé ici est de développer une méthode de gestion des pannes des commandes de vol dans le cas d'une défaillance partielle des actionneurs, qui va permettre à l'avion de poursuivre en toute sécurité la manœuvre prévue. Une approche de commande en deux étapes est proposée et appliquée à la fois à l'évaluation de la manoeuvrabilité restante et à la conception de structures de commande tolérante aux pannes. Dans le premier cas, une méthode d'évaluation hors ligne des qualités de vol basée sur la commande prédictive est proposée. Dans le second cas, une structure de commande tolérante aux pannes basée sur la commande non linéaire inverse et la réaffectation des actionneurs en ligne est développée. Dans les deux cas, un problème de programmation linéaire quadratique (LQ) est formulé. Différents cas de pannes sont considérés lorsqu'un avion effectue une manoeuvre classique. Trois solveurs numériques sont appliqués aux solutions en ligne et hors ligne des problèmes LQ qui en résultent. / The objective of this thesis is to optimize the use of redundant actuators for a transportation aircraft once some actuators failure occurs during the flight. Here, the fault tolerant ability resulting from the redundant actuators is mainly considered. Different classical concepts and methods related to a fault tolerant flight control channel are first reviewed and new concepts useful for the required analysis are introduced. The problem which is tackled here is to develop a design methodology for fault tolerant flight control in the case of a partial actuator failure which will allow the aircraft to continue safely the intended maneuver. A two stages control approach is proposed and applied to both the remaining maneuverability evaluation and a fault tolerant control structure design. In the first case, an offline handling qualities assessment method based on Model Predictive Control is proposed. In the second case, a fault tolerant control structure based on Nonlinear Inverse Control and online actuator reassignment is developed. In both cases, a Linear Quadratic (LQ) programming problem is formulated and different failure cases are considered when an aircraft performs a classical maneuver. Three numerical solvers are studied and applied to the offline and online solutions of the resulting LQ problems.

Theories are tools with which we critically analyze society‟s structure and understand experiences that are not our own. When theories no longer describe the narratives that they claim, we must update them. This thesis proposes to do just that. The trans theories of J. Halberstam and Jay Prosser will be analyzed using ten autobiographies written by self-identified FTM transsexuals and transmen. Prosser and Halberstam are two well-known theorists in the field of trans theory. J. Halberstam approaches FTM transsexuality from the perspective of a lesbian feminist, who has an understanding of transgender butch individuals. Jay Prosser writes from the perspective of being a FTM transsexual himself. I chose publications by these theorists because they had sections that focused specifically on FTM narratives, and they both used at least two of the autobiographies I use and they were published the same year. Therefore, I found them to be two theoretical frameworks that could be compared on several different levels. The publication dates of the ten narratives range from the 1970s to the early 2000s. Applicability of the theories will be measured by comparing them to what the autobiographers say about their lives. The tool of analysis will be five focus areas that are represented, to some degree, in each narrative. They are: Trying to fit into female roll/body/world; Discovering one is Trans/Coming Out; Acquiring Hormones and Sexual Reassignment Surgery; Trying to Establish/Re-establish a Confident Identity; and Romantic/Sexual Relationships.. I will discuss how helpful each theory is in accordance with these narratives. The aim of this thesis is to assess how well each theorist addresses the issue of transsexual identity. The field of transsexual research is still new and developing, there are many interesting parts of these narratives that can be delved into further. I will conclude by proposing a new trans theoretical view point that would better help one to understand the process of female-to-male transition. What one should take away from this thesis is that despite differences in sex and gender formation, these people are, like the rest of the world, trying to create an identity that they not only feel comfortable with as a representation of themselves, but also one that they can be proud of.

A presente dissertação analisa a implementação do Processo Transexualizador no âmbito do Sistema Único de Saúde (SUS), à luz do ideário do Movimento de Reforma Sanitária e de uma perspectiva histórica da política de saúde pública brasileira, detendo-se nas particularidades do Estado do Rio de Janeiro. Discutem-se alguns aspectos da transexualidade relacionados à esfera pública e à efetiva materialização dos direitos da população LGBT, em particular o acesso à saúde de pessoas transexuais. O recorte temporal compreende o período de 1970, quando se iniciam as primeiras cirurgias de transgenitalização no Brasil, a 2008, ano das portarias que instituíram o referido processo. Como instrumentos e técnicas de investigação qualitativa, foram privilegiados o trabalho de campo e a entrevista semiestruturada, tendo sido entrevistados(as) profissionais que atuaram em instituições de saúde que dispunham de programas voltados especificamente à população transexual no Estado do Rio de Janeiro e usuários(as) atendidos(as) por estas instituições. Diante do cenário de discriminação e estigma, muitas vezes fruto do desconhecimento e de informações deturpadas sobre transexualidade, pretende-se conferir maior visibilidade às demandas por direitos de pessoas transexuais, evidenciando a complexidade de tais demandas, bem como as fragilidades do modelo de atenção à saúde subjacente aos mencionados programas. Pretende-se, ainda, contribuir para o fomento da produção acadêmica do Serviço Social, relativamente limitada nesta área. / This dissertation examines the implementation of the sex reassignment process in the Unified Health System (SUS), from the ideals of the Health Reform Movement and a historical perspective of the Brazilian public health policy, focusing on the particularities of the State of Rio de Janeiro. Some aspects of transsexuality related to the state's sphere and to the effective realization of the rights of the LGBT population are discussed, particularly the access to health care for transsexual people. The time frame covers the period from 1970, when the first reassignment surgeries were started in Brazil, and 2008, when such process was officially instituted. The applied methodology favored, as tools and techniques of qualitative research, were fieldwork and semi structured interview with professionals who have worked in the health institutions that had programs geared specifically to transsexual population in the State of Rio de Janeiro and users of health services of these programs. In this scenario of discrimination and stigma, often the result of ignorance and misrepresented information about transsexuality, it is intended to give greater visibility to the health demands of transsexuals, showing the complexity of such demands as well as the weaknesses of the model of healthcare behind the aforementioned programs. It is also intended to contribute to the fomentation of academic production of Social Work, which is relatively limited in this area.

Objectif et problématique : Le transsexualisme ainsi que son traitement par la transformation hormonale et chirurgicale (THC) sont des phénomènes au carrefour de toutes les limites : corps, subjectivité, société, psyché… Face au constat de cette complexité, partant des approches structurale et déconstructiviste, cette thèse a pour objectif de définir et proposer une nouvelle approche « transversale » du transsexualisme, et dans la continuité de celle-ci de l’identité sexuée, avec les éléments de la théorie psychanalytique que sont la pulsion sexuelle de mort, la troisième topique de l’Inconscient clivé, la réalisation pulsionnelle par la perception et la clinique de l’extrême et de l’ordinaire.Matériel et méthode : Sur un échantillon de six sujets transsexuels masculin vers féminin ayant actuellement recours ou ayant eu recours à la THC depuis plus de deux ans, nous avons analysé les résultats à un entretien clinique inspiré de l’approche structurale, aux tests projectifs du rorschach et du TAT selon l’école projectiviste de Paris Descartes, ainsi qu’au MMPI-2. Nos trois hypothèses sont que la problématique identitaire transsexuelle renverrait à la pulsion sexuelle de mort, à l’Inconscient amential et à la clinique de l’extrême, puis que la THC aurait le potentiel d’agir telle une réalisation pulsionnelle par la perception.Résultats : Les résultats valident les deux premières hypothèses sur la pulsion sexuelle, l’Inconscient amential et la clinique de l’extrême, mais pas celle sur la réalisation pulsionnelle par la perception.Conclusion et discussion : Ces résultats montrent l’intérêt d’une approche transversale pour appréhender un phénomène limite comme le sont le transsexualisme et l’identité sexuée qui se voient éclairés par les éléments théoriques transversaux proposés (pulsion sexuelle de mort, Inconscient amential et clinique de l’extrême). Concernant, les effets de la THC au niveau psychique, il semble nécessaire de poursuivre notre investigation, la réalisation pulsionnelle par la perception ayant été validée chez un de nos deux sujets ayant terminé la THC depuis plus de deux ans, mais exclue chez l’autre. / Objective and problematic: the transsexualism as well as its treatment by the hormonal and surgical sex reassignment are phenomena in the crossroads of all the limits: body, subjectivity, society, psyche … In front of this complexity, running with the approaches structural and deconstructivist, this thesis has for objective to define and to propose a new "transversal" approach of the transsexualism, and in the continuity of this one of the sexed identity, with the elements of the psychoanalytical theory that are the sexual death drive, the third topical of the splited Unconscious, the impulsive realization by the perception and the clinical of the extreme and the ordinary.Material(Equipment) and method: on sample of six subjects transsexuals male to female resorting at present or having resorted to the sex reassignment for more than two years, we analyzed the results in a clinical interview inspired by the structural approach, in the projective tests of the rorschach and the TAT according to the projectivist method of Paris Descartes, as well as in the MMPI-2. Our three hypotheses are that the transsexual identity problematic would send back to the sexual death drive, to the amential Unconscious and to the clinical of the extreme, then that the sex reassignment would have the potential to act such as an impulsive realization by the perception.Results: the results validate both first ones hypothesis on the sex drive, the amential Unconscious and the clinical of the extreme, but not that one on the impulsive realization by the perception.Conclusion and discussion: these results show the interest of a transversal approach to dread a phenomenon borderline as are it the transsexualism and the sexed identity which see each other lit by the transversal theoretical proposed elements (sexual death drive, amential Unconscious and private hospital of the extreme). Concerning, the effects of psychic sex reassignment at the level, it seems necessary to pursue our investigation, the impulsive realization by the perception has been validated to one of our subject but excluded to the other one.

This work wishes to present the problem of trans-sexuality and its treatment, especially its most controversial part, i.e. hormonal and surgical reassignment. It brings the information about disease as such, shows treatment possibilities and points out also certain ethical problems which the patient faces, especially when he is a believer. Second part of this work studies the life quality of trans-persons and brings author?s research within a small sample group of transsexual believers. The last part of this work presents summary of results.

A presente dissertação analisa a implementação do Processo Transexualizador no âmbito do Sistema Único de Saúde (SUS), à luz do ideário do Movimento de Reforma Sanitária e de uma perspectiva histórica da política de saúde pública brasileira, detendo-se nas particularidades do Estado do Rio de Janeiro. Discutem-se alguns aspectos da transexualidade relacionados à esfera pública e à efetiva materialização dos direitos da população LGBT, em particular o acesso à saúde de pessoas transexuais. O recorte temporal compreende o período de 1970, quando se iniciam as primeiras cirurgias de transgenitalização no Brasil, a 2008, ano das portarias que instituíram o referido processo. Como instrumentos e técnicas de investigação qualitativa, foram privilegiados o trabalho de campo e a entrevista semiestruturada, tendo sido entrevistados(as) profissionais que atuaram em instituições de saúde que dispunham de programas voltados especificamente à população transexual no Estado do Rio de Janeiro e usuários(as) atendidos(as) por estas instituições. Diante do cenário de discriminação e estigma, muitas vezes fruto do desconhecimento e de informações deturpadas sobre transexualidade, pretende-se conferir maior visibilidade às demandas por direitos de pessoas transexuais, evidenciando a complexidade de tais demandas, bem como as fragilidades do modelo de atenção à saúde subjacente aos mencionados programas. Pretende-se, ainda, contribuir para o fomento da produção acadêmica do Serviço Social, relativamente limitada nesta área. / This dissertation examines the implementation of the sex reassignment process in the Unified Health System (SUS), from the ideals of the Health Reform Movement and a historical perspective of the Brazilian public health policy, focusing on the particularities of the State of Rio de Janeiro. Some aspects of transsexuality related to the state's sphere and to the effective realization of the rights of the LGBT population are discussed, particularly the access to health care for transsexual people. The time frame covers the period from 1970, when the first reassignment surgeries were started in Brazil, and 2008, when such process was officially instituted. The applied methodology favored, as tools and techniques of qualitative research, were fieldwork and semi structured interview with professionals who have worked in the health institutions that had programs geared specifically to transsexual population in the State of Rio de Janeiro and users of health services of these programs. In this scenario of discrimination and stigma, often the result of ignorance and misrepresented information about transsexuality, it is intended to give greater visibility to the health demands of transsexuals, showing the complexity of such demands as well as the weaknesses of the model of healthcare behind the aforementioned programs. It is also intended to contribute to the fomentation of academic production of Social Work, which is relatively limited in this area.