La reconnaissance moléculaire constitue l'une des questions fondamentales les plus discutées dans le domaine de la chimie supramoléculaire. Cette thèse présente la conception, la synthèse et l'étude des propriétés de capsules hélicoïdales auto-organisées par repliement d'oligoamides aromatiques. Ces récepteurs sont constitués d'une chaîne oligomérique se repliant en hélice et comprenant une séquence d'unités codant pour des diamètres différents. Le repliement de l'oligomère donne naissance à une cavité pouvant accueillir des molécules invitées. La grande modularité des séquences, permettant une évolution contrôlée des structures des foldamères, donne lieu à la reconnaissance sélective et anticipée de substrats d'intérêts biologiques. Le phénomène d'encapsulation a été mis en évidence en solution par spectroscopie RMN et CD, et dans le solide par diffraction des rayons X.

L'analyse chimique est indispensable à de nombreuses étapes de la mise au point et dusuivi des procédés de retraitement des combustibles nucléaires usés, de la gestion des déchetsnucléaires, ou encore de l'optimisation des combustibles du futur. Le cycle global d'analysecomprend généralement plusieurs étapes de séparations chimiques longues, manuelles etdifficiles à mettre en oeuvre en raison de leur confinement en boite à gants. Il apparaîtaujourd'hui nécessaire de proposer des solutions innovantes et viables dans le butd'automatiser ces étapes mais aussi de réduire le volume de déchets radioactifs en fin de cycleanalytique. Une solution est alors la conception de plateformes analytiques miniaturiséesautomatisées et jetables.L'objectif de cette thèse est de concevoir un système miniaturisé alternatif à lapremière étape actuelle d'analyse des combustibles usés séparant, par chromatographied'échange d'ions, l'Uranium et le Plutonium des autres éléments constituant l'échantillon. Cesystème doit permettre à la fois de conserver les performances analytiques du processusactuel, de réduire drastiquement l'exposition des expérimentateurs par l'automatisation, ainsique le volume de déchets produits en fin de cycle analytique. Ainsi, la séparation a étéimplantée sur un microsystème jetable en plastique (COC), au design spécialement adapté àl'automatisation : un lab-on-CD.Le prototype développé intègre une micro-colonne séparative d'échange d'anionsremplie d'un polymère monolithique dont la synthèse in-situ ainsi que la fonctionnalisation desurface ont été optimisées spécifiquement pour la séparation voulue. Le développement duprotocole de séparation adapté à ces micro-colonnes a été réalisé à l'aide d'un outil desimulation de l'élution des différents éléments d'intérêts. Cet outil permet de prévoir lagéométrie de la colonne (section et longueur) afin d'obtenir, en fonction de l'échantillon, desfractions de collecte de l'Uranium et du Plutonium pures.Finalement, le prototype actuel est capable de conduire simultanément 4 séparationsde façon automatisée et permet de réduire à la fois le nombre de manipulations, le tempsd'analyse mais aussi de diviser approximativement par 1000 le volume des déchets liquidesgénérés.

[Truncated abstract] Failure to fully reconstitute the immune system is a common clinical problem in HIV patients who were severely immunodeficient before responding to combination antiretroviral therapy (CART). This can manifest as a deficiency in the number or function of CD4+ T-cells and occurs most often in patients who had a nadir CD4+ T-cell count below 100/μl when CART was commenced. Observational studies of large cohorts of HIV patients, such as the D:A:D study, have demonstrated that patients with low CD4+ T-cell counts have increased rates of death compared with patients who have normal CD4+ T-cell counts. Furthermore, individual case studies suggest that impaired recovery of pathogen-specific immune responses during CART is associated with opportunistic infections or disease progression. This thesis addresses possible causes of deficiencies in CD4+ T-cell number or function in HIV patients who were very immunodeficient prior to treatment and are responding (virologically) to CART. Firstly, the role of the thymus in producing naive CD4+ T-cells and the effects of persistent immune activation on the recovery of CD4+ T-cell numbers were assessed in patients with either low or high CD4+ T-cell counts after long-term CART. ... Proportions of antigen presenting cell (APC) subpopulations were examined in HIV patients with low or high CMV-specific CD4+ T-cell responses after long-term CART. HIV patients had significantly lower proportions of plasmacytoid dendritic cells (pDC) than HIV-negative controls. Furthermore, the proportions of pDC were positively correlated with CMV-specific CD4+ T-cell responses in HIV patients. Proportions of myeloid dendritic cells (mDC) were significantly higher in HIV patients than controls, and were also increased in patients with low CMV-specific CD4+ T-cell responses. Proportions of M-DC8+ dendritic cells or CD14+ monocytes did not differ between patients and controls, nor were they associated with CMV-specific CD4+ T-cell responses. Quantitation of cytokine (interferon-α, tumour necrosis factor-α, interleukin (IL) -12, IL-23, IL-15, IL-18 and IL-10) mRNA in unstimulated, purified populations of the APC described above revealed few significant differences between patients with low or high CD4+ T-cell IFN-γ responses to CMV. The only notable difference was the slight elevation of IL-15 mRNA levels in patients compared to controls. Since patients in the high responder group had the highest levels of IL-15 mRNA, this association may reflect the anti-apoptotic properties of IL-15. These studies provide valuable insights into the causes of persistent CD4+ T-cell deficiency and dysfunction in HIV patients on CART and may lead to better monitoring and treatments.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Thesis (Ph.D.) -- University of Texas Southwestern Medical Center at Dallas, 2006. / Not embargoed. Vita. Bibliography: 181-228.

O aumento da expectativa de vida da população mundial tem se associado com uma maior prevalência de doenças neurodegenerativas. A Doença de Alzheimer (DA) é a doença neurodegenerativa mais comum e a principal causa de demência em indivíduos com mais de 60 anos, sendo caracterizada por um declínio progressivo na memória e função mental dos pacientes. Esses sintomas são acompanhados por alterações histopatológicas no cérebro desses indivíduos, incluindo a presença de uma grande quantidade de placas senis, formadas pela deposição do peptídeo beta-amiloide (Aβ), e de emaranhados neurofibrilares formados pela hiperfosforilação da proteína Tau. Estudos indicam que a deposição do Aβ é uma das principais responsáveis pelo desenvolvimento da DA, causando dano neuronal através da ativação de várias vias pró-apoptóticas e dando origem aos sintomas de demência típicos dessa doença. Até o momento, não existem tratamentos eficazes para o combate à DA, de forma que a maior parte das intervenções farmacológicas é destinada apenas ao tratamento de alguns de seus sintomas. A proteína translocadora (TSPO) se localiza em pontos de contato entre as membranas mitocondriais interna e externa e está relacionada com o transporte de colesterol para o interior da mitocôndria e com a regulação da esteroidogênese e da apoptose. Estudos mostram que ligantes da TSPO apresentam efeitos neuroprotetores em diferentes modelos experimentais de lesão cerebral e doenças neurodegenerativas. Especificamente em relação à DA, um estudo indicou que o 4’-clorodiazepam (4’-CD), um ligante da TSPO, apresenta efeitos neuroprotetores em um modelo animal dessa doença, sendo um possível candidato para o seu tratamento. Dessa forma, o objetivo desse estudo foi verificar o efeito neuroprotetor do 4’-CD em diferentes modelos in vitro de toxicidade induzida pelo Aβ, além de seus efeitos sobre o desenvolvimento de neurônios hipocampais. Inicialmente, demonstramos que o 4’-CD reduziu a morte celular de células SH-SY5Y expostas a um modelo de toxicidade induzida pela administração de Aβ. Esses efeitos estiveram associados com a redução da expressão da proteína pró-apoptótica Bax e com um aumento da expressão da survivina, uma proteína anti-apoptótica. A expressão das proteínas Bcl-xl e procaspase-3, por outro lado, não foi alterada pelos tratamentos. Posteriormente, estudamos os efeitos neuroprotetores do 4’-CD contra a toxicidade induzida pela administração do Aβ em culturas organotípicas de hipocampo. Nesses experimentos, foi demonstrado que o 4’-CD reduz a morte celular de culturas organotípicas de hipocampo expostas ao Aβ através de um aumento na expressão da enzima SOD, sem alterar, no entanto, a expressão das proteínas Akt e procaspase-3. Por fim, foi avaliado o efeito do 4’-CD sobre o desenvolvimento de culturas primárias de neurônios hipocampais de camundongos machos e fêmeas. Foi observado que as culturas de neurônios hipocampais das fêmeas apresentaram um desenvolvimento mais rápido do que as dos machos. O 4’-CD acelerou a maturação e aumentou a ramificação neurítica dos neurônios hipocampais dos machos, mas não exerceu qualquer efeito sobre os neurônios das fêmeas. Em suma, foi observado que o 4’-CD apresenta efeitos neuroprotetores contra o Aβ em células SH-SY5Y e em culturas organotípicas do hipocampo, apresentando-se como um fármaco em potencial para o tratamento da DA. Além disso, foi observado que o 4’-CD exerceu um efeito dependente do sexo sobre o desenvolvimento de culturas primárias de neurônios hipocampais, estimulando o desenvolvimento e a ramificação neurítica de neurônios hipocampais de machos, mas não de fêmeas. / The increase in life expectancy of the world population has been associated with a higher prevalence of neurodegenerative diseases. The Alzheimer’s Disease (AD) is the most common neurodegenerative disorder and the main cause of dementia among people over 60 years, being characterized by a progressive decline in the memory and mental function of the patients. These symptoms are associated with histopathological changes in the brain of these patients, including the presence of senile plaques, formed by the deposition of amyloid-beta (Aβ), and neurofibrillary tangles, which are related to the hyperphosphorylation of Tau protein. Studies indicate that Aβ deposition is a major contributor to AD progression, promoting neuronal damage through the activation of different pro-apoptotic pathways and giving rise to the typical dementia symptoms of this disease. To date, there are no effective treatments for AD, so that most of the pharmacological intervention is intended for the treatment of some of its symptoms. The translocator protein (TSPO) is located in contact sites between the outer and the inner mitochondrial membranes and is involved in the cholesterol transport into the mitochondria and in the regulation of steroidogenesis and apoptosis. Studies show that TSPO ligands present neuroprotective effects in different experimental models of brain injury and neurodegenerative diseases. Specifically regarding AD, a study indicated that 4’-chlorodiazepam (4’-CD), a TSPO ligand, is neuroprotective in an animal model of this disease, being a possible candidate for its treatment. Therefore, the aim of this study was to evaluate the neuroprotective effect of 4’-CD in different experimental models of Aβ- induced neurotoxicity in vitro, as well as its effects on the development of hipocampal neurons. First, it was demonstrated that 4’-CD decreased the cell death of SH-SY5Y cells exposed to the Aβ. This effect was associated with the inhibition of the Aβ-induced upregulation of Bax, a pro-apoptotic protein, and downregulation of survivin, a prosurvival protein. On the other hand, the expression of Bcl-xl and procaspase-3 was not change by the treatments. After, it was studied the neuroprotective effects of 4’-CD against Aβ in organotypic hipocampal cultures. In these experiments, it was shown that 4’-CD decreases the cell death of organotypic hippocampal slices exposed to the Aβ by increasing the protein expression of SOD, but without changing the expression of Akt and procaspase-3. Finally, due to the importance of the processes of neuronal development and maturation in the regeneration of CNS after injury, it was evaluated the effect of 4’-CD on the development of primary hippocampal neurons of male and female mice. It was observed that female primary hippocampal neurons presented an increased rate of development than male neurons. 4’-CD stimulated the development and increased the neuritic branching of male but not from female neurons. In summary, it was observed that 4’-CD presented a neuroprotective effect against Aβ in SH-SY5Y cells and in rat organotypical hippocampal slices, presenting itself as a promising agent for the treatment of AD. Also, it was observed that 4’-CD modulates the development of hippocampal neurons in a sex-dependent manner, stimulating the development of male but not from female cells.

The significance of capacity development programmes, as key driver for sustaining development goals, is anchored in all international fora. This research complements and extends our present understanding of the contribution of capacity development approaches to development and achievement of the SDGs. This is achieved by critically assessing the impact of capacity development initiatives sponsored by the World Bank. This thesis has focused on analysis of implementation strategies and critical assessment of the impact of the projects using multidisciplinary approach, utilising a range of quantitative and qualitative methods. It provides a sound empirical basis for assessing the complexities of these projects. This empirical investigation has identified a wide range of disparities of implementation strategies utilised for capacity development initiatives among the major international players (World Bank and UNDP). These findings clearly indicate that there is no single strategy for the implementation of capacity development initiatives. Thus, based on empirical evidence, as well as a critical review of the literature, the study proposes a model for achieving critical sustainable capacity development based on broad and long-term strategies; input, process, output, and outcome which defines the appropriateness of policies and practices that support sustainable development. It is concluded that capacity development initiatives are relevant and essential ensuring national development and sustainable results. The recommendations include the focus on individual, organisational, and societal factors when planning, developing and adopting strategies for implementing all government/national programmes.

Made available in DSpace on 2015-07-22T20:50:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-04-29. Added 1 bitstream(s) on 2015-08-11T03:25:40Z : No. of bitstreams: 1 Publico-062a.pdf: 992519 bytes, checksum: 41750ff4e6e38ad6517d6c7541049844 (MD5). Added 1 bitstream(s) on 2015-08-11T03:25:40Z : No. of bitstreams: 2 Publico-062a.pdf: 992519 bytes, checksum: 41750ff4e6e38ad6517d6c7541049844 (MD5) Publico-062b.pdf: 1669897 bytes, checksum: 5ca81b8b1f9a75f45902de9ce4bc36f7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O GB vírus C (GBV-C) está constituído por uma fita única de RNA de polaridade positiva e pertence à família Flaviviridae. Possui uma seqüência e organização genómica parecida ao vírus da hepatite C, (HCV). A infecção pelo GBV-C não foi associada a nenhuma patologia, embora, na co infecção com o HIV, tenha sido associada a uma sobrevida maior e retardo no desenvolvimento da imunodeficiência. O efeito benéfico do GBV-C parece ser mediado por alterações na resposta imune celular; contudo, os possíveis mecanismos para explicar esse efeito ainda não foram esclarecidos. Neste trabalho investigamos a freqüência e características genotípicas assim como o impacto da infecção pelo GBV-C nos indivíduos infectados pelo HIV-1. No primeiro manuscrito examinamos os conhecimentos descritos na literatura referentes à coinfecção e propusemos algumas hipóteses para explicar esses efeitos. Posteriormente, descrevemos a taxa de infecção, a prevalência, incidência e características genotípicas do GBV-C nesta população. Assim, uma considerável freqüência de infecção pelo GBV-C foi observada e a análise filogenética dos isolados de GBV-C mostraram ser do genótipo 1 e 2. Foi observada também uma correlação inversa entre a carga viral do GBV-C e a carga viral do HIV na inclusão e um ano depois, assim como uma correlação positiva, mas não significativa, entre a carga viral do GBV-C e a contagem de linfócitos T CD4+. Finalmente, avaliamos o efeito da viremia pelo GBV-C na ativação celular em recém infectados pelo HIV-1. Os pacientes foram agrupados em GBV-C viremicos e não virémicos e foram avaliados para a contagem de linfócitos T, marcadores de ativação celular e carga viral do GBV-C e HIV-1. Foram realizadas análises de univariada e multivariada para identificar variáveis associadas com ativação celular. Demonstramos que a viremia pelo GBV-C foi correlacionada com uma diminuição da ativação celular nos indivíduos HIV positivos e este efeito mostrou se independente da carga viral do HIV. Assim, esta associação entre a replicação do GBV-C e menor ativação celular pode explicar, pelo menos em parte, a proteção conferida pelo GBV-C na progressão da doença nos indivíduos infectados pelo HIV-1. / GB virus C (GBV-C) is a single stranded positive sense RNA virus, which is a member of the Flaviviridae. It has a close sequence homology and genomic organization to hepatitis C virus (HCV). No disease has been associated with GBV-C infection but coinfection with human immunodeficiency virus (HIV) leads to improved morbidity and mortality for the HIV infected subjects. The mechanism of the beneficial effect of GBV-C appears to be mediated by alterations in the cellular immune response. In this study we investigated the frequency and genotyping characteristics as well as the impact of the GBV-C infection among recently HIV-1 infected individuals. In the first manuscript we examined the current knowledge concerning this co-infection and developed hypotheses to explain its effects. Subsequently, we described the rate of infection, the prevalence, incidence and genotypic GBV-C characteristics in this population. In that regard, a considerable frequency of GBV-C infection was observed and the phylogenetic analysis of the GBVC isolates revealed the predominance of genotypes 1 and 2. Also, it was observed an inverse correlation between GBV-C load and HIV-1 load at the enrollment and after one year of follow-up, and a positive, but not statistically significant, correlation between GBV-C load and CD4+ T lymphocyte counts. Finally, we have investigated the effect of GBV-C viremia on T cell activation in early HIV-1-infection. The volunteers were enrolled into two groups: GBV-C viremic and non viremic, all co-infected with HIV-1. They were evaluated for T cell counts, cellular activation markers, GBV-C RNA detection, and HIV-1 viral load. Non-parametric univariate and multivariate analyses were carried out to identify the variables associated with cellular activation. We demonstrated that the GBV-C viremia is correlated with a lower T cell activation in HIV-1-infected individuals and this effect was independent of HIV-1 viral load. The association between GBV-C replication and lower T-cell activation may explain, at least in part, the protection conferred by this virus against disease progression to immunodeficiency in HIV-1-infected patients. / TEDE / BV UNIFESP: Teses e dissertações

O aumento da expectativa de vida da população mundial tem se associado com uma maior prevalência de doenças neurodegenerativas. A Doença de Alzheimer (DA) é a doença neurodegenerativa mais comum e a principal causa de demência em indivíduos com mais de 60 anos, sendo caracterizada por um declínio progressivo na memória e função mental dos pacientes. Esses sintomas são acompanhados por alterações histopatológicas no cérebro desses indivíduos, incluindo a presença de uma grande quantidade de placas senis, formadas pela deposição do peptídeo beta-amiloide (Aβ), e de emaranhados neurofibrilares formados pela hiperfosforilação da proteína Tau. Estudos indicam que a deposição do Aβ é uma das principais responsáveis pelo desenvolvimento da DA, causando dano neuronal através da ativação de várias vias pró-apoptóticas e dando origem aos sintomas de demência típicos dessa doença. Até o momento, não existem tratamentos eficazes para o combate à DA, de forma que a maior parte das intervenções farmacológicas é destinada apenas ao tratamento de alguns de seus sintomas. A proteína translocadora (TSPO) se localiza em pontos de contato entre as membranas mitocondriais interna e externa e está relacionada com o transporte de colesterol para o interior da mitocôndria e com a regulação da esteroidogênese e da apoptose. Estudos mostram que ligantes da TSPO apresentam efeitos neuroprotetores em diferentes modelos experimentais de lesão cerebral e doenças neurodegenerativas. Especificamente em relação à DA, um estudo indicou que o 4’-clorodiazepam (4’-CD), um ligante da TSPO, apresenta efeitos neuroprotetores em um modelo animal dessa doença, sendo um possível candidato para o seu tratamento. Dessa forma, o objetivo desse estudo foi verificar o efeito neuroprotetor do 4’-CD em diferentes modelos in vitro de toxicidade induzida pelo Aβ, além de seus efeitos sobre o desenvolvimento de neurônios hipocampais. Inicialmente, demonstramos que o 4’-CD reduziu a morte celular de células SH-SY5Y expostas a um modelo de toxicidade induzida pela administração de Aβ. Esses efeitos estiveram associados com a redução da expressão da proteína pró-apoptótica Bax e com um aumento da expressão da survivina, uma proteína anti-apoptótica. A expressão das proteínas Bcl-xl e procaspase-3, por outro lado, não foi alterada pelos tratamentos. Posteriormente, estudamos os efeitos neuroprotetores do 4’-CD contra a toxicidade induzida pela administração do Aβ em culturas organotípicas de hipocampo. Nesses experimentos, foi demonstrado que o 4’-CD reduz a morte celular de culturas organotípicas de hipocampo expostas ao Aβ através de um aumento na expressão da enzima SOD, sem alterar, no entanto, a expressão das proteínas Akt e procaspase-3. Por fim, foi avaliado o efeito do 4’-CD sobre o desenvolvimento de culturas primárias de neurônios hipocampais de camundongos machos e fêmeas. Foi observado que as culturas de neurônios hipocampais das fêmeas apresentaram um desenvolvimento mais rápido do que as dos machos. O 4’-CD acelerou a maturação e aumentou a ramificação neurítica dos neurônios hipocampais dos machos, mas não exerceu qualquer efeito sobre os neurônios das fêmeas. Em suma, foi observado que o 4’-CD apresenta efeitos neuroprotetores contra o Aβ em células SH-SY5Y e em culturas organotípicas do hipocampo, apresentando-se como um fármaco em potencial para o tratamento da DA. Além disso, foi observado que o 4’-CD exerceu um efeito dependente do sexo sobre o desenvolvimento de culturas primárias de neurônios hipocampais, estimulando o desenvolvimento e a ramificação neurítica de neurônios hipocampais de machos, mas não de fêmeas. / The increase in life expectancy of the world population has been associated with a higher prevalence of neurodegenerative diseases. The Alzheimer’s Disease (AD) is the most common neurodegenerative disorder and the main cause of dementia among people over 60 years, being characterized by a progressive decline in the memory and mental function of the patients. These symptoms are associated with histopathological changes in the brain of these patients, including the presence of senile plaques, formed by the deposition of amyloid-beta (Aβ), and neurofibrillary tangles, which are related to the hyperphosphorylation of Tau protein. Studies indicate that Aβ deposition is a major contributor to AD progression, promoting neuronal damage through the activation of different pro-apoptotic pathways and giving rise to the typical dementia symptoms of this disease. To date, there are no effective treatments for AD, so that most of the pharmacological intervention is intended for the treatment of some of its symptoms. The translocator protein (TSPO) is located in contact sites between the outer and the inner mitochondrial membranes and is involved in the cholesterol transport into the mitochondria and in the regulation of steroidogenesis and apoptosis. Studies show that TSPO ligands present neuroprotective effects in different experimental models of brain injury and neurodegenerative diseases. Specifically regarding AD, a study indicated that 4’-chlorodiazepam (4’-CD), a TSPO ligand, is neuroprotective in an animal model of this disease, being a possible candidate for its treatment. Therefore, the aim of this study was to evaluate the neuroprotective effect of 4’-CD in different experimental models of Aβ- induced neurotoxicity in vitro, as well as its effects on the development of hipocampal neurons. First, it was demonstrated that 4’-CD decreased the cell death of SH-SY5Y cells exposed to the Aβ. This effect was associated with the inhibition of the Aβ-induced upregulation of Bax, a pro-apoptotic protein, and downregulation of survivin, a prosurvival protein. On the other hand, the expression of Bcl-xl and procaspase-3 was not change by the treatments. After, it was studied the neuroprotective effects of 4’-CD against Aβ in organotypic hipocampal cultures. In these experiments, it was shown that 4’-CD decreases the cell death of organotypic hippocampal slices exposed to the Aβ by increasing the protein expression of SOD, but without changing the expression of Akt and procaspase-3. Finally, due to the importance of the processes of neuronal development and maturation in the regeneration of CNS after injury, it was evaluated the effect of 4’-CD on the development of primary hippocampal neurons of male and female mice. It was observed that female primary hippocampal neurons presented an increased rate of development than male neurons. 4’-CD stimulated the development and increased the neuritic branching of male but not from female neurons. In summary, it was observed that 4’-CD presented a neuroprotective effect against Aβ in SH-SY5Y cells and in rat organotypical hippocampal slices, presenting itself as a promising agent for the treatment of AD. Also, it was observed that 4’-CD modulates the development of hippocampal neurons in a sex-dependent manner, stimulating the development of male but not from female cells.

This diploma thesis analyses instances of the initial position of the information focus, the rheme, in present-day English. Initial placement of the rheme is found in structures that go against the principle of end-focus. The aim of this work is to identify which clause constituents, and under what circumstances may function as the initial rheme. The initial section of the theoretical part deals with a description of word-order, and the basic concepts of functional sentence perspective, such as communicative dynamism. Then it continues with the rheme and the factors affecting FSP, and, particularly, with cases where initial rheme occurs. The practical part contains analysis of examples of initial rheme in different genres and text types, including oral communication. The frequency of occurrence is summarized at the end of the work.