Made available in DSpace on 2015-05-14T13:16:39Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1196115 bytes, checksum: 0025bcb835c1d455b4efe0621ecbb6c3 (MD5) Previous issue date: 2010-10-01 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Studies regarding the teaching work in Brazil point out to a frame of precarious work conditions, low wages, full-time journey, social discredit and devaluation. Thus, the present study had as general objective to analyze the work activity of elementary public teaching female teachers from João Pessoa PB, in the perspective of learning the possible subjective mobilization of these workers in a work adverse quotidian. Specifically, we seek to learn the elements of the conditions and forms of the teachers work organization; we identify the regulation strategies of this activity and analyze the competences which the teachers utilize to accomplish their activities. In a way to assist this analysis, we have fallen back mainly upon the theoretical-methodological contributions, guarding its differences, from the Activity Ergonomics and the Work Psychodynamics, operated under the ergological perspective. The operationalization of this study has been developed from the articulation between different techniques which have made possible to emerge the wealth of human relations and the industrial activity in the work situations. To achieve that, we have utilized activity systematic observations, the counterpart instruction technique and mainly the constitution of Research Amplified Communities (CAPs). The analysis of the materials produced has been accomplished following the speech dialogic perspective, from suggestions of the bakhtinian thought. Twenty-five teachers have participated of the research, one man and twenty-four women, all of them proceeding from three elementary public schools, between the ages 21 and 60 years old. From the analysis of the materials produced, we have realized that the main reasons for the insertion and the professional formation of these workers in the mastership are deeply marked by the social relations of gender and class. We have observed the naturalization of the teaching work as a female work , what seems to have implications in the way of performing such work. Regarding the public policies which have been implanted in schools, we have realized that the teachers see them in general in a negative, limited way, since they do not change effectively the reality in schools. The teachers have related that there is not a previous discussion about the implantation of government programs in schools, what many times, instead of favoring their work, has generated overload, what makes difficult the accomplishment of their activities. Concerning the work conditions, we have observed that they are inadequate to the development of the work. The teachers complain, for instance, that the classrooms have inadequate acoustic and ventilation. Besides, they feel paralyzed by the lack of material resources in the exercise of their autonomy. We have verified a discrepancy between prescribed work and real work. The teachers see themselves before a prescription unable to foresee the diversity of situations which they face in the quotidian of schools, thus developing diverse ways of activity regulation, dealing with and inventing different manners of articulating the work, in order to guaranteeing the accomplishment of their activity. We have also learned the ingenuity of sensible intelligence mobilized by the workers in the development of their activities. We have verified yet the living of the nonrecognition of the teachers work by the society and the schools directions, in spite of the signaling of the recognition from students and colleagues. We have come to realize, however, that despite the difficulties found in the work situations, the teachers, through the use of their sensitiveness and creativity, develop diverse ways of activity regulation, giving new forms to the work and creating different ways of articulating themselves to it. / Estudos referentes ao trabalho docente no Brasil apontam para um quadro de condições de trabalho precárias, baixos salários, duplas jornadas de trabalho, desprestígio e desvalorização social. Assim, o presente estudo teve como objetivo geral analisar a atividade de trabalho de professoras da primeira fase do ensino público fundamental do município de João Pessoa-PB, na perspectiva de apreender a possível mobilização subjetiva dessas trabalhadoras em um cotidiano adverso de trabalho. Especificamente, buscamos identificar os elementos das condições e formas de organização do trabalho das professoras; apreender as estratégias de regulação dessa atividade e analisarmos as competências que as professoras utilizam para realizar suas atividades. De forma a subsidiar esta análise, recorremos principalmente às contribuições teórico-metodológicas, resguardando as suas diferenças, da Ergonomia da Atividade, da Psicodinâmica do Trabalho, operadas sob a perspectiva ergológica. A operacionalização desse estudo foi desenvolvida a partir da articulação entre diferentes técnicas que possibilitaram emergir a riqueza da atividade industriosa nas situações de trabalho. Para isso, utilizamos observações sistemáticas da atividade, a técnica de instrução ao sósia e principalmente a constituição de comunidades ampliadas de pesquisa (CAPs). A análise dos materiais produzidos foi realizada seguindo a perspectiva dialógica do discurso, a partir das sinalizações do pensamento bakhtiniano. Participaram da pesquisa vinte e cinco professores, sendo um homem e vinte quatro mulheres, todos provenientes de três escolas do ensino público fundamental, com idade variando de 21 a 60 anos. A partir das análises dos materiais produzidos, percebemos que os principais motivos da inserção e formação profissional dessas trabalhadoras no magistério são profundamente marcados pelas relações sociais de gênero e classe. Observamos a naturalização do trabalho docente como um trabalho feminino , o que parece ter implicações na forma de se exercer tal ofício. Com relação às políticas públicas que vêm sendo implantadas nas escolas, percebemos que as professoras as vêem em geral de forma negativa e/ou limitadas, já que estas não mudam efetivamente a realidade das escolas. As docentes relatam que não há uma discussão prévia sobre a implantação dos programas governamentais nas escolas, o que muitas vezes, ao invés de favorecer seu trabalho, tem gerado sobrecarga, o que dificulta a realização de suas atividades. No que tange às condições de trabalho, observamos que estas são inadequadas para o desenvolvimento do trabalho. É visível a desvalorização expressa nos baixos salários e as professoras se queixam que as salas de aula têm acústica e ventilação inadequadas. Além disso, elas se sentem tolhidas pela falta de recursos materiais e no exercício de sua autonomia. Verificamos uma discrepância entre o trabalho prescrito e o trabalho real. As professoras se vêem diante de uma prescrição incapaz de prever a diversidade de situações com as quais se deparam no cotidiano das escolas, e assim desenvolvem diversas formas de regulação da atividade, antecipando e inventando diferentes maneiras de articular-se ao trabalho, a fim de garantir a realização de sua atividade. Apreendemos ainda a engenhosidade da inteligência prática mobilizada pelas trabalhadoras no desenvolvimento de suas tarefas. Verificamos ainda a vivência do não reconhecimento do trabalho das professoras por parte da sociedade, do governo municipal e das direções das escolas, apesar da sinalização do reconhecimento por parte dos alunos e colegas. Percebemos, entretanto, que apesar das dificuldades encontradas nas situações de trabalho, as professoras por intermédio do uso de suas sensibilidades e criatividades, desenvolvem diversas formas de regulação da atividade, dando novas formas ao trabalho e inventando diferentes maneiras de articular-se a ele.

To unravel the mysteries of complex biological processes carried out by biomolecules it is necessary to adopt a multifaceted approach, which involves employing a wide variety of tools both computational and experimental. In order to gain a clear understanding of the function of biomolecules their three dimensional structure is required. X-ray crystallography and Nuclear Magnetic Resonance (NMR) spectroscopy are the only two methods capable of providing high-resolution three-dimensional structure of biomolecules. NMR has the advantage of allowing the study of structure of biomolecules in solution and is better equipped to characterize the dynamics of the protein. Protein structure determination by NMR spectroscopy consists of recombinant expression of isotopically labeled proteins, purification, data collection, data processing, resonance assignment, distance restraint and angular restraint generation, structure calculation and structure validation. Apart from 3D structure determination of biomolecules NMR has become the method of choice for studying transient protein-protein interactions, which are notoriously difficult to study at higher resolution by other methods. Mass spectrometry plays an important role in enabling rapid identification of biomolecules and their modifications. The high sensitivity and resolution mass spectrometry offers makes it the method of choice for studying post-transitional modification of proteins. Use of computers in biology has played an essential role in elucidating those structure function relationships in biomolecules that are not possible to study by experimental techniques. The first chapter of this thesis deals with the introduction of methods used in this study. A brief introduction about the theory of Nuclear Magnetic Resonance (NMR) spectroscopy is given. Protein NMR methods used for structure determination of medium sized proteins are discussed. A part of this chapter discusses about the application of mass spectrometry in biochemistry and the use of tandem MS/MS experiments in identification of proteins and peptide fragments. Finally, the last part of this chapter gives an introduction about the theory of molecular dynamics and techniques used in the post processing of MD trajectories to elucidate the dynamics of proteins. The second chapter of this thesis is concerned with NMR characterization of a novel protein-protein interaction between the glycolytic enzyme Triosephosphate isomerase and the redox protein Thioredoxin. Chemical shift perturbation studies have been done to map the binding interfaces of these proteins. The structure of the complex was then modeled using NMR restraints based docking using the known 3D structure of these proteins. The docked complex reveals crucial insights into the glutathione mediated redox regulation of Triosephosphate isomerase and the role of thioredoxin as a deglutathionylating agent. Enzyme activity assays of Triosephosphate isomerase were done to show the inhibitory effects of s-glutathionylation of Cys217 and the role of thioredoxin as a deglutathionylating agent. The third chapter of the thesis is aimed to address some important issues related to the inhibition of Plasmodium falciparum Triosephosphate isomerase by S-glutathionylation. Oxidative stress induces protein glutathionylation which is a reversible post translational modification consisting of the formation of a mixed disulfide between protein cysteines and glutathione. Mass spectrometric analysis of the kilnetics of glutathionylation along with enzyme activity assays clearly show that gluthionylation of either Cys-13 (situated in the dimmer interface) or Cys-217 (situated in Helix G) can render the enzyme inactive. Molecular dynamics simulations provide a mechanistic basis of inhibition and predict that glutathionylation at Cys217 allosterically induces loop 6 disorder. The fourth chapter of this thesis addresses the stabilizing effect of introduction of a cross-strand disulfide bond across a non-hydrogen bonded position of an antiparallel beta sheet. Multidimensional heteronuclear NMR experiments have been used to get the backbone and side-chain resonance assignments, distance and angular restraints. In addition RDC based restraints have been used to calculate the structure of oxidixed form of L79C, T89C thiroedoxin. The observation of predominantly –RH staple conformation among the NMR ensemble in typical of cross-strand disulfides. The fifth chapter of this thesis deals with the dynamics of thioredoxin using computational methods.In this chapter analysis of known complexes of thiroedoxin was done to determine binding hot spot residues using free energy calculations. The physicochemical basis for the multispecificity of thioredoxin is probed using molecular dynamics simulations. In this chapter it has been shown that conformational selection plays a very important role in thioredoxin target recognition.