Dans la levure S. cerevisiae, la mitose nécessite le positionnement du fuseau mitotique le long de l’axe cellule mère-bourgeon (future cellule fille) afin d‘assurer une bonne ségrégation des chromosomes. Ce phénomène requiert le fonctionnement de deux mécanismes impliquant les protéines Kar9 et Dyn1. Durant la métaphase, Kar9 se positionne de manière asymétrique le long du fuseau mitotique, avec une accumulation notable sur les microtubules qui émanent de l’ancien « spindle pole body » (SPB; l’équivalent du centrosome dans les vertébrés), qui est normalement dirigé vers le bourgeon. Dans le cas d’un défaut d’alignement du fuseau mitotique, un mécanisme appelé « Spindle Position Checkpoint » (SPOC) inhibe la sortie de mitose et la cytokinèse, afin de permettre un réalignement correct du fuseau mitotique. La principale cible de ce checkpoint est une GTPase Tem1. Dans le cas d’alignement correct du fuseau mitotique, Tem1 active une voie de signalisation appelée le « Mitotic Exit Network » (MEN) qui permet de mener à la sortie de mitose et à la cytokinèse. Lors de la transition métaphase/anaphase Tem1 se positionne asymétriquement sur les SPBs jusqu’à se concentrer majoritairement sur l’ancien SPB. Des données récentes ont montré que des composants du MEN, Tem1 inclus, sont également impliqués dans la régulation de la localisation de la protéine Kar9 à l’SPB, et dans l’établissement d’une polarité correcte des SPBs durant la métaphase. En effet, Kar9 se positionne plus symétriquement dans le cas des mutants du MEN que dans le type sauvage, ce qui engendre des problèmes d’orientation du fuseau et de ségrégation des SPBs. Nous cherchons à élucider comment l’activité du MEN régule la localisation de Kar9 et l’orientation du fuseau mitotique en métaphase alors que les fonctions du MEN liées à la sortie de mitose restent bloquées jusqu’à la télophase. Nous avons émis l’hypothèse que les modifications post-traductionnelles de Tem1 pourraient jouer un rôle dans la régulation du MEN. Il a été montré que les résidus Y40 et Y45 sont phosphoryles in vivo. Afin de disséquer le rôle de ces résidus nous les avons mutés en phénylalanines. Ces mutations peuvent complémenter la létalité induite par la délétion de TEM1, suggérant que ce mutant conserve les fonctions essentielles de Tem1. Par ailleurs, la cinétique de progression du cycle cellulaire du mutant est la même que celle du type sauvage, signifiant que la perte de phosphorylation sur Tem1 ne semble pas agir sur la sortie de mitose. De plus, l’allèle mutant n’affecte pas la localisation aux SPBs de Tem1 ni celle de sa « GTPase-activating protein » Bub2/Bfa1 durant le cycle cellulaire. Bien que l’activité GTPasique de la protéine Tem1-Y40F,Y45F soit réduite in vitro, les mutations ne causent pas des défauts de SPOC in vivo et le mutant répond efficacement au mauvais alignement de fuseau mitotique en s’arrêtant en anaphase. Tous ces résultats nous suggèrent que la perte de phosphorylation de Tem1 n’affecte pas les fonctions de fin de mitose de cette GTPase. Par contre, nous avons découvert que la phosphorylation de Tem1 est requise pour la localisation asymétrique de Kar9 sur les SPBs, ainsi que pour l’alignement correct du fuseau mitotique durant la métaphase (la distribution de Kar9 est plus symétrique dans les cellules TEM1-Y40F,Y45F et que le fuseau mitotique n’est pas aligné correctement). Nous cherchons alors à trouver quelle kinase phosphoryle Tem1 et régule son activité. Les kinases potentielles sont la protéine Swe1 (la seule vraie kinase phosphorylant les tyrosines dans la levure) ainsi que la kinase Mps1 (kinase qui contrôle la duplication des SPBs). Nous développons actuellement des outils nous permettant de vérifier l’implication de ces deux candidats. Mots clés : Tem1, Kar9, cycle cellulaire, Mitotic Exit Network (MEN), Spindle Position Checkpoint (SPOC), phosphorylation on tyrosines. / In the budding yeast Saccharomyces cerevisiae a faithful mitosis requires positioning of the mitotic spindle along the mother-bud axis to ensure proper chromosome segregation. This is achieved by two distinct but functionally redundant mechanisms that require the APC (adenomatous polyposis coli)-like protein Kar9 and dynein (Dyn1), respectively. During metaphase, Kar9 localizes asymmetrically on the mitotic spindle, with a prominent accumulation on astral microtubules emanating from the old spindle pole body (SPB – i.e. the yeast equivalent of the centrosome) that is normally directed towards the bud. In case of spindle misalignment, a surveillance mechanism called Spindle Position Checkpoint (SPOC) inhibits mitotic exit and cytokinesis, thereby providing the time necessary to correct spindle alignment. The main target of the SPOC is the small GTPase Tem1, which activates a signal transduction cascade called Mitotic Exit Network (MEN) that drives cells out of mitosis and triggers cytokinesis. Tem1 is localized at SPBs, with an increasingly asymmetric pattern during the progression from metaphase to anaphase, when Tem1 is concentrated on bud-directed old SPB. Recent data have implicated MEN components also in the regulation of Kar9 localization at SPBs and in setting the right polarity of SPBs inheritance during metaphase. In particular, Kar9 localizes more symmetrically in MEN mutants than in wild type cells and this leads to spindle orientation and SPB inheritance defects (i.e. with the new SPB being oriented towards the bud). A key question emerging from these data is how MEN activity is regulated to promote proper Kar9 localization and spindle positioning in metaphase, while being restrained until telophase for what concerns its mitotic exit and cytokinetic functions. We hypothesised that Tem1 post-translational modifications might be relevant for this control and for this reason we have been focusing on the role of Tem1 phosphorylation. Tem1 was found in a wide phosphoproteomic study to be phosphorylated on two tyrosines (Y40 and Y45) located at its N-terminus. We constructed a non-phosphorylatable mutant, TEM1-Y40F,Y45F, where the two phosphorylated tyrosines were mutated to phenylalanine. This mutant allele was able to rescue the lethality caused by TEM1 deletion, suggesting that it retains all its the essential functions. The kinetics of cell cycle progression of TEM1-Y40F,Y45F cells was similar to that of wild type cells, suggesting that lack of Tem1 phosphorylation is unlikely to affect mitotic exit. In addition, the TEM1-Y40F,Y45F allele did not affect the SPB localization of Tem1 and its regulatory GTPase-activating protein Bub2/Bfa1 during the cell cycle. Moreover, although the Tem1-Y40F,Y45F mutant protein showed reduced GTPase activity in vitro, it did not cause SPOC defects in vivo and could efficiently respond to spindle mispositioning. Altogether, these results suggest that lack of Tem1 phosphorylation does not affect the late mitotic functions of the GTPase. In contrast, we found that Tem1 phosphorylation is required for Kar9 asymmetry at SPBs and proper spindle positioning during metaphase. Indeed, TEM1-Y40F,Y45F cells display a more symmetric pattern of Kar9 distribution at SPBs in this cell cycle stage, as well as spindle position and orientation defects. We are currently investigating if Tem1 phosphorylation also regulates the pattern of SPB inheritance. Finally, an important question that we are trying to answer is “what is the kinase that phosphorylates Tem1?” The best candidates are the wee1-like kinase Swe1, which is the only true tyrosine kinase of budding yeast, and Mps1, a dual-specificity protein kinase controlling SPB duplication. While we are developing specific tools to study Tem1 phosphorylation and ultimately identify its promoting kinase, we gained preliminary data suggesting that both kinases might be involved in spindle positioning.

Propõe-se um controlador difuso Takagi-Sugeno com índice de desempenho H \'INFINITO\' para regulagem da marcha lenta em motores de ciclo Otto. Obtém-se uma representação difusa do motor de ciclo Otto operando em marcha lenta. Esse modelo é utilizado para a síntese do controlador. O controlador difuso com desempenho H \'INFINITO\' é projetado para rejeitar o efeito de distúrbios de cargas externas sobre a rotação do motor em regime de marcha lenta. As ações de controle, posição da borboleta de aceleração e ponto de ignição, são limitadas a uma faixa de operação específica para a marcha lenta. O controlador projetado apresenta ação de controle por realimentação de estados. Os ganhos de realimentação para cada regra do controlador difuso são obtidos a partir de um problema de otimização formulado através de LMIs. / It is proposed an Otto cicle engine idle speed H \'INFINITE\' Takagi-Sugeno fuzzy control. It is presented an Otto cicle engine idle speed fuzzy model. This model is then used to control design. The H \'INFINITE\' fuzzy control is designed to reject external load disturbance effect at idle speed engine rotation. A specifc idle speed operation range is defined to both actions control, throttle plate position and spark advance. A state feedback control is designed. A LMI optmization problem is used to find the state feedback gains at each fuzzy control rule.

O presente estudo está inserido na Linha de Pesquisa Educação, Linguagem e Psicologia do Programa de Pós-Graduação da Faculdade de Educação da Universidade de São Paulo (USP). Considerando o ensino da língua escrita como a função primeira da escola a fim de formar leitores e escritores proficientes, justifica-se o Ciclo de Alfabetização como âmbito privilegiado de atenção, sobretudo quando se tem em vista os resultados insatisfatórios das pesquisas de avaliação do desempenho escolar. Nessa perspectiva, o presente trabalho tem como objetivo o estudo das práticas pedagógicas nas classes de 1º ao 3º ano do Ensino Fundamental. Partindo da hipótese de que, no contexto escolar, pode haver oscilação nas propostas de ensino com diferentes implicações para o processo de aprendizagem, o foco da investigação está nas práticas e intervenções docentes. Assim, o estudo constitui-se pela observação de seis turmas do Ciclo de Alfabetização em uma escola da rede pública estadual paulista, na cidade de São José dos Campos, em dois diferentes momentos (2014 e 2015), com o intuito de registrar as propostas de trabalho no ensino da Língua Portuguesa a partir de quatro eixos: a natureza da atividade, a natureza da demanda feita ao aluno, a natureza linguística da proposta e a natureza interacional na dinâmica de produção da turma. Os dados foram coletados com base nas concepções dialógica de língua (Bakhtin), e interacionista de aprendizagem (Piaget e Vygostky); mais especificamente, na alfabetização como processo constituinte da pessoa pela progressiva aproximação do aluno com as culturas do escrito em situações de reflexão e efetiva comunicação social (Geraldi, Ferreiro, Teberosky, Lerner, Weisz e Sanchez). As conclusões do estudo apontam para o predomínio de atividades de escrita em uma perspectiva notacional, nas quais prevalecem o objeto e a razão do escrever em detrimento da definição de interlocutores. Em outras palavras, a escrita na escola configura-se mais como objeto de aprendizagem do que como prática comunicativa. O ensino, por sua vez, aparece geralmente centrado no professor, isto é, com poucas oportunidades de reflexão e interação entre alunos. No conjunto dos dados, comprovou-se a oscilação entre turmas do mesmo ano e também na progressão de cada turma, o que põe em evidência a difusão das atividades de língua portuguesa em planos de trabalho nem sempre coerentes com o perfil das turmas ou com as diretrizes de ensino. A análise desses dados remete à necessidade de se repensar os planos de ensino e a progressão do trabalho na escola, além de subsidiar projetos de formação docente. / The present study is inserted in the Research Line \"Education, Language and Psychology\" of the Post-Graduation Program of the Faculty of Education of the University of São Paulo (USP). Considering the teaching of written language, the schools primary function in a perspective of forming proficient readers and writers, warrants the Literacy Cycle as a privileged scope of attention, mainly when bearing in mind the unsatisfactory results of evaluation researches about the school performance. In this regard, the present research aimed to study pedagogical practices in 1st to 3rd grades of elementary schools. Since in the school environment there can be oscillation in the teaching proposals with different implications for the learning process, the focus of this investigation is on teachers practices and interventions. Therefore, this study was formed by the observation of six groups of the Literacy Cycle in a state school in São José dos Campos São Paulo, in two different moments (2014 and 2015), in order to register forms of teaching the Portuguese Language, based on four paths: the nature of the activity, the nature of the demand directed to the student, the linguistic nature of the proposal and the interactional nature in the class production dynamics. The data were collected based on dialogical concepts of the language (Bakhtin) and interactionist concept of the learning process (Piaget and Vygotsky); more specifically, in literacy as a human process resulted from the progressive approach of the student with the written cultures in situations of reflection and real social communication (Geraldi, Ferreiro, Teberosky, Lerner, Weisz and Sanchez). This research concluded the predominance of activities based on a notational perspective, in which the object and the reasons to write prevail over the definition of interlocutors. In other words, in this research writing at school was more an object of learning than a communicative practice. Teaching, in turn, was centered in the teacher, that is, students had few opportunities to reflect or interact among themselves. All this information proves the oscillation amid classes of the same grade, as well as in each groups progress, which emphasizes the diffusion of Portuguese activities in plannings not always coherent with the groups profile or with the official teaching directions. The analysis of these information brings the urge to rethink about teaching plannings and progression of the work at school, besides, it subsidizes projects of professional development for teachers.

This thesis project is focused on the study of the signaling module formed by the NIMA-related protein Nek6, Nek7, and Nek9 and their function during early mitosis, with particular interest in centrosome separation and maturation. Nek9/Nercc1 was identified by Dr. Joan Roig. Nek9 is expressed in all cell lines and tissues studied is inactive during interphase while during mitosis is activated through phosphorylation by Plk1 which is in fact able to bind Nek9 and subsequently phosphorylates Nek9 on its activation loop. During mitosis Nek6 and Nek7 bind the C-terminal of Nek9. Once active, Nek9 can phosphorylate Nek6 and Nek7, thus activating them. Active Nek9 localizes at centrosome, suggesting that Nek9/Nek6-7 has important functions in the organization of microtubules during cell division. Confirming this idea, it has been shown that the microinjection of anti-Nek9 module induces arrest in prometaphase with disorganized spindle structures and misaligned chromosomes, or leads to abnormal mitosis resulting in aneuploidy. In the same direction, interference with the function of Nek7 or Nek6 leads to abnormal mitotic progression and spindle formation. We described how the Nek9/Nek6-7 module could provide a link connecting Plk1 and Eg5 in the context of centrosome separation. we analyzed the effects of Plk1, Eg5, Nek9, Nek6 or Nek7 down-regulation by RNAi on the extent of separation of duplicated centrosomes in prophase cells and we observed how this downregulation was affecting centrosome separation. We determine whether the activation of Nek9 or Nek6 could induce centrosome separation trasfecting cells with the active form of these two kinases; a considerable amount of cells that were in interphase shown separate centrosome demonstrating that Nek9/Nek6 are sufficient to induce centrosome separation. To test whether active Nek9 and Nek6 exerted their effect through the regulation of Eg5 we simultaneously transfected the cells with Eg5 siRNAs and we completely lost the centrosome separation described above. We demonstrated by immunofluorescence that the key event during centrosome separation was the recruitment of Eg5 at centrosomes and that the down-regulation of Plk1, Nek6, Nek7 or Nek9 resulted in prophase cells with unseparated centrosomes because Eg5 was not properly recruited. To prove whether the phosphorylation on Ser-1033 controls the accumulation of Eg5 to centrosomes and centrosome separation during early mitosis we transfected cells with wild type Eg5 or Eg5 S1033A; the wild type form of the kinesin was able to localize at centrosome and rescue the normal phenotype while Eg5 S1033A was not able to localize and resulted in cells delayed in mitosis. Plk1, the Nek9 activator, is involved in the regulation of centrosome maturation during early mitosis. Centrosome maturation refers to the process through which centrosomes increase size and microtubule nucleation activity and requires the accumulation of γ-TuRC complexes at centrosome. This recruitment depends on Nedd1 that acts as γ-Tubulin targeting factor. Plk1 depletion prevents accumulation of Nedd1 at centrosome. Our experiments show the importance of Nek9 in the regulation of centrosome maturation downstream of Plk1. Depletion of Nek9 by siRNA determined a decrease of γ-Tubulin and Nedd1 at centrosome. Further we investigated the upstream role of Plk1 depleting Plk1 and trasfecting active Nek9 and it was able to rescue the normal phenotype. Nek9 can interact with Nedd1 during mitosis and phosphorylates it provoking its accumulation at centrosome. The no-phosphorylable form of Nedd1 was not able to accumulate at centrosome and support the accumulation of γ-Tubulin there, determining a delay of the cells in prometaphase. Our results show that Nek9 is the link between Plk1 activity and the recruitment of Nedd1 to the centrosome and that the pathway formed by Plk1/Nek9/Nedd1 can be a key element in the control of mitotic centrosome maturation.

Existeixen moltes evidències que indiquen que les diferents funcions descrites de p21(Cip1) es deuen, en certa part, a la seva localització intracel·lular. Mentre que la p21(Cip1) nuclear inhibeix la proliferació cel·lular, la p21(Cip1) citoplasmàtica és capaç de regular la supervivència i la mobilitat cel·lular. El fet que les funcions de p21(Cip1) en cada un dels compartiments cel·lulars tinguin papers oposats fa que sigui d'un gran interès l'estudi dels canvis de localització de p21(Cip1) i els mecanismes que regulen aquesta translocació, així com les possibles vies que p21(Cip1) pugui fer servir per a localitzar-se en els diferents compartiments cel·lulars.En aquesta tesi s'han realitzat dos estudis diferenciats, però amb un objectiu comú ja que tots dos es centren en l'anàlisi dels mecanismes reguladors de la localització cel·lular de p21(Cip1). En la primera part d'aquesta tesi, observem la importància de la fosforilació de p21(Cip1) per part de la proteïna cinasa C (PKC). Aquesta fosforilació té lloc en el residu Ser153, molt proper a la senyal de localització nuclear (NLS) de la p21(Cip1) i es capaç de regular la localització cel·lular de p21(Cip1). Aquests resultats, juntament amb altres treballs, demostren que la fosforilació de p21(Cip1) afavoreix la seva localització en el citoplasma. Diferents aproximacions experimentals ens van permetre observar com aquesta fosforilació inhibeix la unió entre p21(Cip1) i CaM. D'aquest primer treball se'n deriva un model de regulació de la localització cel·lular de p21(Cip1) en el qual la unió a CaM i la fosforilació per PKC tenen papers oposats. D'una banda, la unió de p21(Cip1) a CaM inhibeix la seva fosforilació per part de PKC i afavoreix la localització de p21(Cip1) en el nucli. D'altra banda, la fosforilació de p21(Cip1) en el residu Ser153 indueix una localització citoplasmàtica de p21(Cip1). Treballs posteriors ens van permetre observar com la sortida de p21(Cip1) del nucli cap al citoplasma també està regulada. Després del dany al DNA, els nivells cel·lulars de p21(Cip1) incrementen, especialment en el nucli, per tal d'assegurar una aturada del cicle cel·lular. Observem com en resposta al dany al DNA, p21(Cip1) s'acumula no només en el nucleoplasma de les cèl·lules sinó que també s'acumula en el nuclèol. En les cèl·lules danyades els components nucleolars es troben desorganitzats i el nuclèol perd els contactes amb l'embolcall nuclear i presenta unes estructures esfèriques en el seu interior. La p21(Cip1) present en les estructures esfèriques del nuclèol està en un equilibri dinàmic amb la p21(Cip1) del nucleoplasma i la presència de p21(Cip1) en aquestes estructures correlaciona amb una inhibició de l'export p21(Cip1) cap al citoplasma. Aquests resultats donen suport a l'existència d'una via d'export de proteïnes nuclears a través del nuclèol, semblant a la que intervé en l'export de ribosomes, la qual es veuria afectada amb la desestructuració dels nuclèols en resposta al dany cel·lular. A més, amb els resultats obtinguts no descartem la possibilitat que p21(Cip1) pugui ser modificada en el nuclèol ja que en resposta al dany al DNA també s'hi localitzen altres proteïnes implicades en diferents vies de modificació post-traduccional. Així doncs, l'acumulació nuclear de p21(Cip1) en resposta al dany al DNA no es deu únicament a un increment de la seva transcripció sinó que aquesta acumulació també es deguda a la inhibició de la sortida de p21(Cip1) cap al citoplasma a través del nuclèol.En conclusió, tant l'entrada com la sortida de la p21(Cip1) del nucli és un mecanisme altament regulat que farà que la localització de p21(Cip1) pugui variar en diferents situacions fisiològiques de la cèl·lula. Aquest fet és de gran importància per al correcte funcionament cel·lular ja que com hem descrit anteriorment, p21(Cip1) és una proteïna amb funcions oposades depenent de la seva localització intracel·lular: oncogènica al citoplasma i supressora de tumors al nucli. / It is well known that p21(Cip1) is a protein with a dual function in oncogenesis depending mainly on its intracellular localization: tumor suppressor in the nucleus and oncogenic in the cytoplasm. The importance of p21(Cip1) cellular localization indicates that it has to be precisely regulated.On one side we observed the importance of p21(Cip1) phosphorylation by PKC inducing its cytoplasmic localization. PKC phosphorylates p21(Cip1) at Ser 153 and when phosphorylated, p21(Cip1) can not bind to CaM. From this study we conclude that CaM and PKC have an opposite role in the regulation of p21(Cip1) localization: CaM binding to p21(Cip1) prevents its phosphorylation by PKC at Ser153 and consequently allows its nuclear localization; while when phosphorylated at Ser153, p21(Cip1) is located at the cytoplasm.On the other side the export of p21(Cip1) from the nucleus to the cytoplasm is also regulated. After DNA damage, p21(Cip1) increases and accumulates in the nucleus to ensure cell cycle arrest. We observed that after DNA damage p21(Cip1) accumulates not only in the nucleoplasm but also in the disrupted nucleoli. In damaged cells the nucleolar components are disorganized and nucleoli have lost their contacts with the nuclear envelope and appear with spherical structures inside. The nucleolar p21(Cip1) forms a dynamic equilibrium between the nucleolus and the nucleoplasm and correlates with the inhibition of p21(Cip1) nuclear export. This result proves the existence of a nucleolar export route to the cytoplasm for p21(Cip1) similar to the one described for the ribosome export. Moreover, the results obtained suggested that p21(Cip1) could be modified in the nucleolus in response to DNA damage as different proteins involved in post-translation modifications also localize in the nucleoli after the damage. Thus, after DNA damage, p21(Cip1) accumulates in the nucleus due to an increase in its transcription and due to an inhibition of its export to the cytoplasm.All this results together indicate the importance of p21(Cip1) localization depending on the cellular context and that its localization is precisely regulated by different pathways.

Flotillin-1 és una proteïna associada a membrana plasmàtica implicada en processos de trànsit de vesícules, reordenació del citoesquelet i transducció de senyals. Estudis previs en el laboratori han demostrat que Flotillin-1 és capaç de translocar-se a nucli en resposta a un estímul mitogènic i afavorir la proliferació de diverses línies cel·lulars. Els mecanismes mitjançant els quals provoca aquests efectes són desconeguts i objecte del present estudi.D'una banda demostrem que Flotillin-1 és un factor regulador de la cinasa Aurora B, una proteïna que intervé en el control de la mitosi i més concretament en el anaphase checkpoint. El knock-down de Flotillin-1 provoca events mitòtics aberrants, acompanyats del descens tant en l'expressió d'Aurora B com de la seva activitat mesurada com els nivells de fosforilació de la histona H3. Flotillin-1 interacciona amb Aurora B i evita la seva degradació per la via del proteasoma.D'altra banda, Flotillin-1 interacciona amb el receptor transmembrana Notch1, implicat en nombrosos processos de regulació de proliferació, diferenciació, apoptosi, etc. Flotillin-1 regula la localització subcel·lular de Notch1 així com la seva capacitat com activador transcripcional. La depleció o mutació de Flotillin-1 dificulta l'entrada de Notch1 a nucli i l'expressió dels gens diana de les famílies Hes/Hrt. En conjunt, es presenta a Flotillin-1 com una proteïna capaç d'actuar a diferents nivells i regular processos i vies de senyalització cel·lular que li confereixen un paper com a regulador de la proliferació cel·lular. / Flotillin-1 is a protein associated to plasma membrane involved in vesicle trafficking, cyotskeleton reorganization and signal transduction. Previous findings in our laboratory has shown that Flotillin-1 is able to translocate the nucleus under mitogenic stimulus and increase proliferation rates of several cell lines. The mechanisms of action are unknown and object of the present study.First, we show that Flotillin-1 is a regulator factor of the mitotic kinase Aurora B, a protein involved in control of mitosis and, specifically, in the anaphase checkpoint. The knock-down of Flotillin-1 causes aberrant mitotic events, decrease in Aurora B levels and its activity, measured as protein levels of phosporilated histone H3. Flotillin-1 interacts with Aurora B and avoid its degradation by the proteasome pathway.In addition, Flotillin-1 interacts with the transmembrane receptor Notch1, involved in many regulatory processes of proliferation, differentiation, apoptosis, etc. Flotillin-1 regulates the subcellular localization of Notch1 and its activity as transcriptional activator. The mutation or depletion of Flotillin-1 difficult the entry of Notch1 in the nucleus and the expression of its target genes Hes/ HRT. Overall, Flotillin-1 is a protein capable of acting at different levels, processes and signaling pathways in order to be a regulator of cell proliferation.

El tema objecte d'aquesta tesi doctoral prové d'una necessitat personal, d'anys d'experiència en l'àmbit educatiu i de diferents estudis realitzats. El plantejament sorgeix del perquè la pràctica educativa continua essent tant difícil, complexa i problemàtica. Aquesta qüestió dóna pas a l'intent de trobar alternatives per a canviar i facilitar la tasca docent, i alhora millorar l'aprenentatge de l'alumnat que hi ha a les escoles.En els darrers cursos en els que s'ha portat a terme una tasca assessora en diferents centres educatius, i s'han atès demandes del professorat referides a alumnes amb necessitats educatives especials, ha provocat la necessitat de cercar estratègies de treball que facilitin al professorat l'atenció a la diversitat. L'orígen d'aquesta tesi es planteja en la recerca de noves estratègies o alternatives a oferir als docents per fer front a l'alumnat que hi ha a les aules. La principal aspiració és la de conseguir un estudi assumible personalment i per als qui va dirigit, amb resultats pràctics i funcionals. En aquest sentit apareix la necessitat d'acotar el treball i es pren la decisió d'emmarcar l'estudi en l'Etapa d'Educació Infantil. La pretensió darrera és la de fer un treball d'observació i anàlisi de la pràctica educativa a l'aula, per tal de recollir totes aquelles actuacions del professorat que afavoreixen l'atenció a la diversitat. Per poder-ho dur a terme, es disposa de l'oferiment de diferents centres educatius de la població de Santa Coloma de Gramenet, on alguns professionals es mostren oberts i disposats a col.laborar en aquesta investigació.El fet de centrar l'estudi i recerca d'estratègies en l'àmbit de l'aula, no pretén oblidar la necessitat de tenir molt present la inclusió d'aquestes estratègies a nivell global de centre, tant a nivell del Projecte Curricular de Centre com d'aspectes organitzatius i de funcionament. L'elecció però de centrar la investigació a l'aula ve motivada per la necessitat de resoldre el "dia a dia" de l'acció educativa, essent però molt conscient de que, per a un bon resultat, cal que hi hagi un plantejament i recolçament de tot el centre educatiu al darrera. En definitiva la pràctica educativa és una interacció de diversos sistemes i entre ells l'aula, amb els seus participants, la seva organització i el seufuncionament propis.Del contrast d'informacions que s'obté i l'anàlisi realitzat sobre els diferents elements d'interacció a l'aula (interrelació alumne/professor, els continguts i tasques, i la dimensió temporal del procés d'ensenyament i aprenentatge), els interessos dels nens i nenes de 3 a 6 anys, l'opinió de les famílies i dels professors, respecte l'aprenentatge dels seus fills i alumnes respectivament, s'elabora una proposta d'estratègies amb models d'intervenció per a poder atendre la diversitat a l'aula.L'estructura de la tesi està formada per dos grans blocs. El primer (corresponent al volum I de l'original) es troba dividit en tres parts; a la Part I es realitza tot un estudi teòric, respecte a l'educació infantil, el currículum, els plantejaments de la reforma i la intervenció educativa, com a fonamentació de la posterior recerca. En la Part II s'exposa tot el procés de la recerca: el disseny de l'estudi, la mostra, la justificació i descripció dels instruments elaborats per a la recollida d'informació, la metodologia utilitzada, les dades i resultats obtinguts, i un recull d'estratègies en relació a aquests. En la IPart III s'elabora una proposta d'estratègies didàctiques per l'atenció a la diversitat a l'aula, d'alumnes d'educació infantil, oferint una breu presentació o explicació per cadascuna d'elles i exemplificant-les amb una projecció a la realitat educativa que serveixi de model per al professorat. Per finalitzar, es realitzen les conclusions finals així com una proposta de possibles treballs de recerca a dur a terme en un futur. També es fa esment de tota la bibliografia i documentació utilitzada. L'altre bloc està integrat per tota la documentació recollida per tal de fonamentar la tesi, i que ocupava els volums II a IV de la tesi.

Cell cycle control by trophic factors has a key role in regulation of cell proliferation in all organisms. Nutrients are one of these important factors needed by cells to reproduce, so very well regulated mechanisms must exist that connect nutrient availability to cell cycle. Hence the importance on studying how exactly nutrient-dependent signaling pathways work. Cln3, the most upstream G1 cyclin in Saccharomyces cerevisiae, is one well demonstrated common effector of multiple nutrient-dependent signaling pathways. Moreover, its role in cell cycle is crucial. So it is a good candidate to regulate cell cycle progression in response to nutrient availability. One important question is to find the protein that could directly modulate Cln3 levels in response to nutrient availability. This protein could play as a nutrient sensor and as a cell cycle regulator at the same time. In the present thesis, Pho85 is founded to be the protein that could run these two highly different tasks, because of its well-characterized properties on sensing phosphate availability and the well-known functions on modulating cell cycle as CDK. The results of the present work clearly demonstrate that when phosphate is present, Pho85 regulates Cln3 levels by increasing the stability of the cyclin through specific phosphorylations, promoting cell cycle progression. Contrary, under phosphate depletion conditions, Pho85 become inactive and Cln3 is rapidly degraded, leading to a cell cycle arrest in order to maintain cell chronological lifespan. / El control del cicle cel•lular per factors tròfics té un paper important en la proliferació cel•lular de tots els organismes. Els nutrients són uns d’aquests factors importants requerits per les cèl•lules per reproduir-se, per tant deuen existir mecanismes molt ben regulats que connecten la disponibilitat de nutrients amb el cicle cel•lular. Per això, l’estudi de com funciona la senyalització cel•lular de nutrients i com afecta a la progressió del cicle és altament rellevant. Cln3, la ciclina de G1 més primerenca a Saccharomyces cerevisiae, és un efector comú de múltiples vies de senyalització de nutrients. A més, el seu paper en el cicle cel•lular és crucial. Per tant aquesta proteïna és una bona candidata per regular la progressió del cicle cel•lular en resposta a la disponibilitat de nutrients. Una qüestió important a resoldre és trobar la proteïna que podria modular directament els nivells de Cln3 depenent de la presència de nutrients. Aquesta proteïna actuaria com a sensor de nutrients i com a reguladora del cicle cel•lular alhora. A la present tesi, es mostra a Pho85 com la proteïna que pot fer aquestes dues tasques, tant per les seves propietats ben conegudes en la detecció de fosfat, com per les seves funcions de CDK modulant el cicle cel•lular. Els resultats d’aquesta tesi demostren clarament que quan el fosfat és present, Pho85 modula els nivells de Cln3 incrementant l’estabilitat de la ciclina mitjançant fosforilacions específiques, promovent la progressió del cicle cel•lular. Per altra banda, sota condicions de manca de fosfat, Pho85 esdevé inactiva i Cln3 és degradada ràpidament, conduint a un arrest del cicle cel•lular per mantenir la longevitat de la cèl•lula. / El control del ciclo celular por factores tróficos tiene un papel importante en la proliferación celular de todos los organismos. Los nutrientes son uno de estos factores importantes requeridos por las células para reproducirse, por lo tanto deben existir mecanismos muy bien regulados que conecten la disponibilidad de nutrientes con el ciclo celular. Por ello, el estudio de cómo funciona la señalización celular de nutrientes y cómo afecta a la progresión del cicle es altamente relevante. Cln3, la ciclina de G1 más temprana en Saccharomyces cerevisia, es un efector común de múltiples vías de señalización de nutrientes. Además, su papel en el ciclo celular es crucial. Por lo tanto esta proteína es una buena candidata para regular la progresión del ciclo celular en respuesta a la disponibilidad de nutrientes. Un tema importante a resolver es encontrar la proteína que podría modular directamente los niveles de Cln3 dependiendo de la presencia de nutrientes. Esta proteína actuaría como sensor de nutrientes y como reguladora del ciclo celular. En la presente tesis, se muestra a Pho85 como la proteína que puede hacer estas dos tareas, tanto por sus propiedades bien conocidas en la detección de fosfato, como por sus funciones de CDK modulando el ciclo celular. Los resultados de esta tesis demuestran claramente que cuando el fosfato está presente, Pho85 modula los niveles de Cln3 incrementando la estabilidad de la ciclina mediante fosforilaciones específicas, promoviendo la progresión del ciclo celular. Por otro lado, bajo condiciones de ausencia de fosfato, Pho85 es inactivada y Cln3 se degrada rápidamente, conduciendo a una parada del ciclo celular para mantener la longevidad de la célula.

Submitted by Milena Rubi (milenarubi@ufscar.br) on 2017-02-14T18:20:18Z No. of bitstreams: 1 PINTO_JUNIOR_Marcos_2016.pdf: 74569511 bytes, checksum: 32a68b14280a0775bacb90a6092e5223 (MD5) / Approved for entry into archive by Milena Rubi (milenarubi@ufscar.br) on 2017-02-14T18:20:29Z (GMT) No. of bitstreams: 1 PINTO_JUNIOR_Marcos_2016.pdf: 74569511 bytes, checksum: 32a68b14280a0775bacb90a6092e5223 (MD5) / Approved for entry into archive by Milena Rubi (milenarubi@ufscar.br) on 2017-02-14T18:20:39Z (GMT) No. of bitstreams: 1 PINTO_JUNIOR_Marcos_2016.pdf: 74569511 bytes, checksum: 32a68b14280a0775bacb90a6092e5223 (MD5) / Made available in DSpace on 2017-02-14T18:20:47Z (GMT). No. of bitstreams: 1 PINTO_JUNIOR_Marcos_2016.pdf: 74569511 bytes, checksum: 32a68b14280a0775bacb90a6092e5223 (MD5) Previous issue date: 2016-08-22 / Não recebi financiamento / This research is an exploratory study presenting the Design for Environment - DfE as one of the environmental management practices in Industrial Ecology in the product development cycle. The DfE should examine the entire life cycle of a product, such as its development, manufacture, use and disposal proposing changes to the project in order to minimize its environmental impact. This product design technique makes it possible to achieve the usual goals in their design, such as performance, reliability and cost of manufacturing. These together with environmental goals, for example to reduce environmental damage, reduced use of resources, increasing energy efficiency and material recycling. Thus, there is need for replacement almost obligatory for some product types. However, the rapid replacement of these products, as well as its disposal can create serious environmental problems, by volume, the material in its composition that take a long time to decompose, such as plastic, glass and metal, but especially because of the heavy metals that can compose, are highly damaging to human health. Not being enough, there are still suitable locations for disposal of many products into disuse. The growth of the contingent of consumers who prefer to buy goods and services that respect the nature is one of the factors that drives the treatment of environmental problems, in addition to the rapid popularization of products and obsolescence of some models. Aspects of reduction of natural resources can be developed into a company that include one Lean Manufacturing environment that aims at a reduction of waste, seeking to improve productivity and quality. This relationship contribute to sustainable development suggesting the existence of an enabling environment for the realization of efforts aimed at reducing or eliminating waste. Through a case study it became clear that DfE practices is related to some Lean tools, as part of its purpose. Also some tools that the company uses environmental exert influence, may be applied in product design to minimize the environmental impact. This work presents the practices observed by a Literature Review Systematic comparison with the case study and the DfE practices that relate to Lean. / Esta pesquisa é um estudo exploratório apresentando o Design for Environment - DfE como sendo uma das práticas de gestão ambiental em Ecologia Industrial no ciclo de desenvolvimento do produto. O DfE deve examinar todo o ciclo de vida de um produto, como seu desenvolvimento, fabricação, uso e disposição final propondo alterações no projeto, de forma a minimizar seu impacto ambiental. Esta técnica de projeto de produto possibilita atingir os objetivos usuais em sua concepção, como desempenho, confiabilidade e custo de manufatura. Estes em conjunto com os objetivos ambientais, por exemplo a redução em danos ambientais, redução do uso de recursos naturais, incremento da eficiência energética e reciclagem de materiais. Desta forma, há necessidade de substituição quase obrigatória para alguns tipos de produto. Todavia, a rápida substituição desses produtos, assim como o seu descarte pode gerar sérios problemas ambientais, pelo volume, pelos materiais em sua composição que demoram muito tempo para se decompor, tais como: o plástico, o vidro e o metal, mas, especialmente, por causa dos metais pesados que pode o compor, são altamente prejudiciais à saúde humana. Não sendo suficiente, ainda faltam locais apropriados para a disposição final de muitos produtos em desuso. O crescimento do contingente de consumidores que preferem comprar produtos e serviços que respeitem a natureza é um dos fatores que impulsiona o tratamento dos problemas ambientais, além da rápida popularização de produtos e obsolescência de alguns modelos. Os aspectos de redução de recursos naturais podem ser desenvolvidos em uma empresa que contemplem um ambiente de Lean Manufacturing que objetiva uma redução de desperdícios, buscando a melhoria da produtividade e da qualidade. Essa relação contribuir para o desenvolvimento sustentável sugerindo a existência de um ambiente propício para a realização de esforços voltados para a redução ou eliminação de desperdícios. Por meio de um estudo de caso foi possível evidenciar que práticas de DfE tem relação com algumas ferramentas de Lean, sendo parte de seu propósito. Também algumas ferramentas que a empresa utiliza exercem influência ambientais, podendo ser aplicadas na concepção do produto para minimização do impacto ambiental. Assim, este trabalho apresenta as práticas observadas por meio de uma Revisão Bibliográfica Sistemática comparando com o caso estudado e as práticas de DfE que se relacionam com o Lean.

Capes / O desenvolvimento de produtos é apontado na literatura e meio empresarial como uma questão desafiadora. Além disso, no atual cenário em que as empresas estão inseridas, discussões sobre o desenvolvimento sustentável é cada vez mais presente e essencial. Neste sentido, a Avaliação do Ciclo de Vida (ACV) é uma ferramenta importante para ajudar a garantir a sustentabilidade adequada por meio da avaliação dos impactos ambientais no desenvolvimento de produtos. Este trabalho tem como objetivo propor uma metodologia para integração da ACV ao processo de desenvolvimento de produto. Para isso, este estudo foi realizado em três fases. Na primeira fase, foi realizado o desenvolvimento teórico dos temas. Na segunda fase, foi realizada uma revisão sistemática da literatura, onde foi possível analisar o estado da arte sobre a consideração da ACV no processo de desenvolvimento de produto (PDP). E, na terceira fase, a estruturação da metodologia para integração da ACV ao PDP foi desenvolvida. A metodologia proposta, foi elaborada com base na ACV e nas fases do processo de desenvolvimento apresentado pela ISO/TR 14062. O objetivo de considerar a ACV no PDP é auxiliar na identificação de potenciais de melhoria ambiental e na definição de metas ambientais, além de, possibilitar o desenvolvimento de novas estratégias para o produto. Deste modo, a metodologia proposta para integração da ACV no PDP é realizada em três macro fases: Pré-integração, Integração e Pós-integração. Essas macro fases são constituídas por quatro fases: escolha do produto de referência, ACV do produto de referência, integração da ACV no PDP e análise da integração da ACV no PDP. Sendo que cada fase apresenta atividades especificas a serem realizadas. Na fase de integração da ACV no PDP são apresentadas ações para integração em cada fase do PDP e a análise dos resultados da integração é realizada por meio de uma matriz de avaliação que considera as categorias de impacto e as fases do ciclo de vida do produto. A estrutura da metodologia proposta é apresentada de modo genérico e flexível o que possibilita sua aplicação em diferentes processos, podendo ser adaptada as características e necessidades das organizações. Desta forma, a metodologia desenvolvida, poderá auxiliar as empresas no processo de desenvolvimento de produtos mais sustentáveis aumentando o nível de competitividade da organização e auxiliando as empresas a identificarem novas oportunidades para os produtos desenvolvidos. / Product development is touted as a challenging issue in academia and industry. In addition, in the current scenario in which companies are inserted, discussions on sustainable development are increasingly present and essential. In this sense, Life Cycle Assessment (LCA) is an important tool to help ensure adequate sustainability by assessing environmental impacts in product development. Therefore, this work aims to propose a methodology for the integration of LCA to the process of product development in the industry. For this, this study was carried out in three phases. In the first phase, the theoretical development of the themes was carried out. In the second phase, a systematic review of the literature was carried out, where it was possible to analyze the art study about the consideration of LCA in the product development process (PDP). And, in the third phase, the structuring of the methodology for integrating LCA into the PDP was developed. The proposed methodology was developed based on the LCA and in the stages of the development process presented by ISO / TR 14062. The objective of considering LCA in the PDP is to help identify environmental improvement potentials and to set environmental goals, besides, to enable the development of new strategies for the product. Thus, the suggested methodology for integrating the LCA into the PDP is carried out in three macro phases: Pre-integration, Integration and Post-integration. These macro phases consist of four phases: choice of the reference product, ACV of the reference product, integration of the ACV in the PDP and analysis of the integration of the ACV in the PDP. Each phase has specific activities to be carried out. In the integration phase of the LCA in the PDP, actions are presented for integration in each phase of the PDP and the analysis of the results of the integration is performed through an evaluation matrix that considers the impact categories and the phases of the product life cycle. The structure of the proposed methodology is presented in a generic and flexible way. This allows its application in different processes, and the characteristics and needs of the organizations can be adapted. In this way, the methodology developed will help companies in the process of developing more sustainable products by increasing the level of competitiveness of the organization and helping companies to identify new opportunities for the products developed.