<p>Megakaryocyte cultures are a strong tool for the in vitro investigation of platelet production in platelet disorders. Peripheral blood derived hematopoietic progenitor cells (PB-HPCs) are the most accessible source of HPCs with high potential to produce mature megakaryocytes in vitro; however, they are present in low numbers making peripheral blood an inefficient source. Additionally, a megakaryocyte culture with an optimized thrombopoietin (TPO) concentration is required which can reliably allow the investigation of suppressive effects of antibodies/plasma from immune thrombocytopenia (ITP) patients. In this study, we developed a megakaryocyte culture with the utilization of human PB-HPCs in an efficient fashion resulting in the production of high purity megakaryocytes in a TPO-dependent manner.</p> <p>The mononuclear fraction was collected from 180 mL of peripheral whole blood and CD34+ cells were isolated by a positive selection yielding the average of 5.5 x 105 ± 2.5 x 105 CD34+ cells (n = 18). Using 96-well tissue-culture plates and seeding 10,000 CD34+ cells/well, the average of 13 experiments in triplicate can be set up utilizing isolated CD34+ in an efficient manner. Capitalizing on a TPO dose-dependent megakaryocyte production experiment, 20 ng/mL was established as the TPO concentration which resulted in the production of mature megakaryocytes without reaching the plateau in megakaryopoiesis response. On day 11 of culture, the expression of megakaryocytic lineage (CD41/61+) and maturation (CD41/61+CD42+) markers peaked at 90.65% and 76.10%. In conclusion, this culture system has broad application for investigation of platelet disorders and drug discovery which can be accessible to all researchers.</p> / Master of Science (MSc)

O exercício da profissão docente requer uma sólida formação, não apenas nos conteúdos científicos da disciplina, mas também nos aspectos correspondentes a sua didática e ao encaminhamento dos diferentes saberes que caracterizam a docência no ensino superior. A formação científica e pedagógica dos professores universitários é um dos fatores básicos na qualidade da universidade, pesquisas apontam que a maioria dos professores possuem lacunas na formação pedagógica. A Universidade de São Paulo USP, lócus desta pesquisa, institui em 1999 o Programa de Aperfeiçoamento de Ensino (PAE) para aprimorar a formação do estudante stricto sensu por meio de duas etapas: Preparação Pedagógica e Estágio Supervisionado em Docência. Dessa maneira o objetivo desse estudo é investigar se o Programa de Aperfeiçoamento de Ensino (PAE) tem-se constituído em espaço formativo para a docência superior a partir da análise das etapas de Preparação Pedagógica e Estágio Supervisionado em Docência. Para tanto, a coleta de dados ocorreu em diferentes momentos. No primeiro momento foram levantados os dados nos Programa de Pós-graduação Stricto Sensu da Faculdade de Educação (humanas), na Escola Politécnica (exatas) e na Escola de Enfermagem (biológicas) do Campus São Paulo Capital, no período de 2001 a 2011. No segundo momento e a partir dos dados coletados, um questionário foi enviado aos pós-graduandos que participaram das duas Etapas do PAE nos Programas investigados. De natureza qualitativa, o procedimento metodológico adotado foi o Estudo de Caso Único Integrado, seguidas da análise documental e análise de conteúdo. As análises apontaram que grande parte dos pós-graduandos, participantes do Programa PAE reconhecem: a ausência da formação pedagógica do professor universitário nos programas de pós-graduação stricto sensu, e identificam contribuições do PAE para o exercício da docência em ambas as etapas (Preparação Pedagógica e Estágio Supervisionado em Docência) possibilitando o aprimoramento e aprofundamento profissional. Com relação à Etapa de Preparação Pedagógica os dados revelam que o conjunto de conferências é a atividade mais procurada pelos alunos, seguida das disciplinas e do núcleo de atividades. Embora mais procurada, percebem o conjunto de conferências como uma proposta pouco efetiva para a formação docente. Quanto à Etapa do Estágio Supervisionado em Docência, os participantes, destacam ser este o verdadeiro momento de formação pedagógica, por proporcionar a reflexão sobre os saberes do campo pedagógico de maneira sistemática, organizada, os aproximando da realidade da sala de aula na graduação. Por fim, a conclusão deste estudo revela que o PAE representa na Universidade de São Paulo, um espaço de formação para a docência, entretanto, não explora todo seu potencial formativo. Ressaltamos a necessidade de sanar as ações isoladas que fragmentam e fragilizam a formação pedagógica do pós-graduando e, ainda, destacamos a urgência da criação de políticas institucionais e diretrizes específicas que tratem dessa formação no Programa PAE e consequentemente nos programas de pós-graduação da Universidade de São Paulo. / The professor professional practice requires a solid education, not only in the scientific discipline content but also in the aspects corresponding to its didactic and in the leading of different knowledge which characterizes teaching in higher education. The scientific and pedagogic education of university professors is one of the ground factors in the quality of the university; researches show that the majority of the professors show gaps in their pedagogic education. The University of São Paulo USP, lócus of this research, established in 1999 the Improvement Teaching Program (ITP) to perfect strictosensustudents education in two stages: Pedagogic Preparation and Supervised Internship in Teaching. As a result, the aim of this study is to investigate whether the Improvement Teaching Program (ITP) has been an educational environment to higher education professors considering the analysis of different stages (Pedagogic Preparation and Supervised Internship in Teaching). In order to do that,data collection occurred in two different instances. In a first instance, data collection was made from 2001 to 2011 in Education College StrictoSensuPost-Graduation Program (Humanities), Polytechnic School (Exact science) and Nursing School (Biological science), São Paulo campus capital. In a second instance, a questionnaire was sent to the students who participated in ITP in the above mentioned teaching unities. Concerning qualitative nature, the adopted methodological procedure was Case Study followed by the documental and content analysis.Analysis show that the great part of post-graduating, taking part in ITP Program acknowledge the lack of pedagogic education of university professor in strictosensupost-graduation programs and identified contributions of ITP for higher education practice in both stages (Pedagogic Preparation and Supervised Internship in Teaching), allowing the professional deepening and improvement. Concerning the Pedagogic Preparation, data reveals that the conference group is the activity which interested students the most, followed by the disciplines and by the activity nucleus. Although it was the one that interested students the most, they realize the conference group as a fairly effective proposal to teaching education. Regarding to Supervised Internship in Teaching, the participants highlighted it as the actual moment of pedagogic education as it provides reflection on the pedagogic field knowledge in a systematic and organized way approaching them to the reality of a classroom in graduation. All in all, the conclusion of this study reveals that ITP represents at University of São Paulo a higher education environment, however, it does not explore its formative potential. It is worth pointing out the need to resolve isolated actions that fragment and undermine the post-graduation pedagogic education, and the urgency of the creation of institutional and specific policies which addresses to this education at ITP Program and consequently in post-graduation programs of University of São Paulo.

Il n'existe à l'heure actuelle pratiquement pas de copolymères greffés à fonctions cationiques et/ou amphotères présentant une dégradabilité intrinsèque. L'objectif de cette thèse est de générer des composés répondant à ces critères à partir d'une méthode de modification chimique par voie anionique de poly(e-caprolactone) (PCL). A cette fin, deux stratégies principales ont été suivies à partir d'un même intermédiaire réactionnel, un macropolycarbanion dérivé de PCL. La première est fondée sur l'emploi du macropolycabanion en tant qu'agent nucléophile qui réagit avec de petites molécules organiques électrophiles. Ces réactions de substitution ont permis l'obtention de copolymères à base PCL possédant entre 10 et 15% de motifs substitués par des groupes cationiques (ammonium, phosphonium), ou hydrosolubilisants (amine, acide carboxylique). Ces squelettes PCL fonctionnalisés peuvent être utilisés pour effectuer des post-modifications (réactions de couplages, dérivation de fonctions). La seconde stratégie utilise le macropolycarbanion en tant que macroamorceur de polymérisation anionique. Ce type de réaction a été appliqué à des dérivés vinyliques (4-VP, N-VP), des dérivés acryliques (DMAEM, MAPEG), des dérivés acrylamides (DMA, DMAPMA) et des dérivés cycliques (NCA d'acides a-aminés) ce qui a permis d'obtenir des copolymères greffés hydrosolubles à chaîne principale PCL. Ces composés ont des structures partiellement dégradables et forment en solution aqueuse des objets de dimensions nanométriques. Enfin, l'utilisation d'un dérivé polyiodé de PCL en tant que macro-polyagent de transfert est abordée ce qui constitue une nouvelle méthode de polymérisation radicalaire contrôlée par transfert dégénératif d'iode permettant l'obtention de structures greffées.

O exercício da profissão docente requer uma sólida formação, não apenas nos conteúdos científicos da disciplina, mas também nos aspectos correspondentes a sua didática e ao encaminhamento dos diferentes saberes que caracterizam a docência no ensino superior. A formação científica e pedagógica dos professores universitários é um dos fatores básicos na qualidade da universidade, pesquisas apontam que a maioria dos professores possuem lacunas na formação pedagógica. A Universidade de São Paulo USP, lócus desta pesquisa, institui em 1999 o Programa de Aperfeiçoamento de Ensino (PAE) para aprimorar a formação do estudante stricto sensu por meio de duas etapas: Preparação Pedagógica e Estágio Supervisionado em Docência. Dessa maneira o objetivo desse estudo é investigar se o Programa de Aperfeiçoamento de Ensino (PAE) tem-se constituído em espaço formativo para a docência superior a partir da análise das etapas de Preparação Pedagógica e Estágio Supervisionado em Docência. Para tanto, a coleta de dados ocorreu em diferentes momentos. No primeiro momento foram levantados os dados nos Programa de Pós-graduação Stricto Sensu da Faculdade de Educação (humanas), na Escola Politécnica (exatas) e na Escola de Enfermagem (biológicas) do Campus São Paulo Capital, no período de 2001 a 2011. No segundo momento e a partir dos dados coletados, um questionário foi enviado aos pós-graduandos que participaram das duas Etapas do PAE nos Programas investigados. De natureza qualitativa, o procedimento metodológico adotado foi o Estudo de Caso Único Integrado, seguidas da análise documental e análise de conteúdo. As análises apontaram que grande parte dos pós-graduandos, participantes do Programa PAE reconhecem: a ausência da formação pedagógica do professor universitário nos programas de pós-graduação stricto sensu, e identificam contribuições do PAE para o exercício da docência em ambas as etapas (Preparação Pedagógica e Estágio Supervisionado em Docência) possibilitando o aprimoramento e aprofundamento profissional. Com relação à Etapa de Preparação Pedagógica os dados revelam que o conjunto de conferências é a atividade mais procurada pelos alunos, seguida das disciplinas e do núcleo de atividades. Embora mais procurada, percebem o conjunto de conferências como uma proposta pouco efetiva para a formação docente. Quanto à Etapa do Estágio Supervisionado em Docência, os participantes, destacam ser este o verdadeiro momento de formação pedagógica, por proporcionar a reflexão sobre os saberes do campo pedagógico de maneira sistemática, organizada, os aproximando da realidade da sala de aula na graduação. Por fim, a conclusão deste estudo revela que o PAE representa na Universidade de São Paulo, um espaço de formação para a docência, entretanto, não explora todo seu potencial formativo. Ressaltamos a necessidade de sanar as ações isoladas que fragmentam e fragilizam a formação pedagógica do pós-graduando e, ainda, destacamos a urgência da criação de políticas institucionais e diretrizes específicas que tratem dessa formação no Programa PAE e consequentemente nos programas de pós-graduação da Universidade de São Paulo. / The professor professional practice requires a solid education, not only in the scientific discipline content but also in the aspects corresponding to its didactic and in the leading of different knowledge which characterizes teaching in higher education. The scientific and pedagogic education of university professors is one of the ground factors in the quality of the university; researches show that the majority of the professors show gaps in their pedagogic education. The University of São Paulo USP, lócus of this research, established in 1999 the Improvement Teaching Program (ITP) to perfect strictosensustudents education in two stages: Pedagogic Preparation and Supervised Internship in Teaching. As a result, the aim of this study is to investigate whether the Improvement Teaching Program (ITP) has been an educational environment to higher education professors considering the analysis of different stages (Pedagogic Preparation and Supervised Internship in Teaching). In order to do that,data collection occurred in two different instances. In a first instance, data collection was made from 2001 to 2011 in Education College StrictoSensuPost-Graduation Program (Humanities), Polytechnic School (Exact science) and Nursing School (Biological science), São Paulo campus capital. In a second instance, a questionnaire was sent to the students who participated in ITP in the above mentioned teaching unities. Concerning qualitative nature, the adopted methodological procedure was Case Study followed by the documental and content analysis.Analysis show that the great part of post-graduating, taking part in ITP Program acknowledge the lack of pedagogic education of university professor in strictosensupost-graduation programs and identified contributions of ITP for higher education practice in both stages (Pedagogic Preparation and Supervised Internship in Teaching), allowing the professional deepening and improvement. Concerning the Pedagogic Preparation, data reveals that the conference group is the activity which interested students the most, followed by the disciplines and by the activity nucleus. Although it was the one that interested students the most, they realize the conference group as a fairly effective proposal to teaching education. Regarding to Supervised Internship in Teaching, the participants highlighted it as the actual moment of pedagogic education as it provides reflection on the pedagogic field knowledge in a systematic and organized way approaching them to the reality of a classroom in graduation. All in all, the conclusion of this study reveals that ITP represents at University of São Paulo a higher education environment, however, it does not explore its formative potential. It is worth pointing out the need to resolve isolated actions that fragment and undermine the post-graduation pedagogic education, and the urgency of the creation of institutional and specific policies which addresses to this education at ITP Program and consequently in post-graduation programs of University of São Paulo.

No description available.

Les cellules lymphoïdes innées (ILCs) sont des populations cellulaires d’identification récente, mais leur rôle in vivo chez l’homme reste mal connu. Dans une 1ère étude, nous avons pu montrer qu’un déficit sévère en NK au cours de déficits immunitaires communs variables est associé à un risque accru de manifestations non infectieuses et infectieuses bactériennes sévères, suggérant un rôle protecteur non redondant des cellules NK lorsque le système immunitaire adaptatif n’est pas fonctionnel. Dans une 2ème étude, nous avons montré que des patients atteints de déficits immunitaires combinés sévères ɣc et JAK3 déficients n’ont pas d’ILCs. Après allogreffe de moelle osseuse, le nombre d’ILCs circulantes reste indétectable, sans manifestation clinique notable associée. Ces résultats sont en faveur d’une redondance des fonctions des ILCs chez l’homme, lorsque les fonctions T et B sont conservées. Nous avons ensuite étudié les modifications phénotypiques et fonctionnelles des cellules NK au cours du purpura thrombopénique immunologique, et observé un défaut de production d’interféron-ɣ par les cellules NK circulantes et une augmentation de la cytotoxicité dépendante des anticorps des cellules NK spléniques. Une inhibition des fonctions des cellules NK par les immunoglobulines polyvalentes est également mise en évidence. Enfin, une étude des ILCs circulantes au cours de la maladie associée aux IgG4 ainsi qu’une revue de la littérature sur l’étude des ILCs au cours des pathologies inflammatoires sont rapportées. En conclusion, l’apparente redondance des ILCs chez l’homme ainsi que leur implication en pathologies inflammatoires en font de potentielles cibles thérapeutiques. / Innate lymphoid cells (ILCs) are recently identified components of the immune system, but their functions in vivo in humans are still elusive. In a first study, we show in patients with common variable immunodeficiency that non-infectious inflammatory complications and severe bacterial infections were more frequent in patients with severe NK cell lymphopenia, indicating potential non-redundant immune functions of NK cells when the adaptive immune response is not optimal. In a second study, we observe that in patients with ɣc and JAK3 severe combined immunodeficiencies, all ILC subsets are absent. After hematopoietic stem cell transplantation, ILCs remain indetectable with no susceptibility to disease, suggesting that ILCs might be redundant and dispensable in humans, if T and B cells functions are preserved. In the second part of this thesis, we study phenotypic and functional modifications of NK cell compartment in primary immune thrombocytopenia. Interferon gamma production by the peripheral blood NK cells of ITP patients is decreased. In contrast, splenic NK cells of ITP patients tend to be more efficient in antibody-dependent cell cytotoxicity. Intravenous polyvalent immunoglobulins lead to the inhibition of blood NK cell activation. Finally, we present the preliminary results of a study investigating the modifications of circulating ILCs in IgG4-related disease, and present an extensive litterature review concerning the role of ILCs in inflammatory diseases. In conclusion, the apparent redundancy of ILCs for protective immunity and their pathogenic role in inflammatory diseases make their targeting in humans for therapeutic purposes particularly promising.

Title: The effect of airway control on stance dynamic stability Objectives: The aim of this thesis is to find out whether and how will the influence of vocal cords modulation be manifested on the dynamic stabilization of the standing body during translational shifts of the supporting surface of different intensities and A-P directions. Thus, building on the findings of Massery et al (2013). Methods: The thesis has the character of qualitative research. The experiment was attended by 23 healthy probands, of which 7 men and 16 women aged 20-40 years. Spirometry was used to test the objectivity of airway airflow during breathing / phoning maneuvers with different vocal cords positioning and dynamic computer posturography using the Neurocom Smart Equi Test System and its Motor Control Test, which evaluated the effectiveness of automatic postural responses. We connected the posturograph with the spirometer using the Kistler accelerometer (type 8766A100BB). The course of the experiment was simultaneously recorded by a camera (GoPro Hero 7). The Smart EquiTest System generated three postural perturbations of different intensity (S - sub treshold, M - threshold, L - saturating) in two directions (anterior translation / posterior translation). The measured data were then processed in the program Neurocom...

No description available.

No description available.

Formal verification of programs can be done with the aid of an interactive theorem prover. The program to be verified is represented in an intermediate language representation inside the interactive theorem prover, after which statements and their proofs can be constructed. This is a process that can be automated to a high degree. This thesis presents a proof procedure to efficiently generate a theorem stating the weakest precondition for a program to terminate successfully in a state upon which a certain postcondition is placed. Specifically, the Poly/ML implementation of the SML metalanguage is used to generate a theorem in the HOL4 interactive theorem prover regarding the properties of a program written in BIR, an abstract intermediate representation of machine code used in the PROSPER project. / Bevis av säkerhetsegenskaper hos program genom formell verifiering kan göras med hjälp av interaktiva teorembevisare. Det program som skall verifieras representeras i en mellanliggande språkrepresentation inuti den interaktiva teorembevisaren, varefter påståenden kan konstrueras, som sedan bevisas. Detta är en process som kan automatiseras i hög grad. Här presenterar vi en metod för att effektivt skapa och bevisa ett teorem som visar sundheten hos den svagaste förutsättningen för att ett program avslutas framgångsrikt under ett givet postvillkor. Specifikt använder vi Poly/ML-implementationen av SML för att generera ett teorem i den interaktiva teorembevisaren HOL4 som beskriver egenskaper hos ett program i BIR, en abstrakt mellanrepresentation av maskinkod som används i PROSPER-projektet.