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Completando 100 anos da descoberta da molestia, a Doenca de Chagas, causada pelo agente etiologico Trypanosoma cruzi, ainda e considerada nas Americas Central e do Sul um problema de saude publica, atingindo mais de 8 milhoes de individuos. Sua transmissao ocorre de distintas maneiras, como por vetores, transfusao sanguinea, transplante de orgaos, acidentes em laboratorios, via oral. O parasita e conhecido por sua heterogeneidade no genotipo e fenotipo, baseado em mutacoes cumulativas em diferentes sub-populacoes do parasita. A cepa CL Brener e uma cepa padrao para pesquisa com diversas peculiaridades interessantes como baixa infectividade em animais, resposta aos tratamentos existentes e seus marcadores geneticos serem estaveis. O clone CL B5 de T. cruzi e originado a partir da cepa CL Brener modificada geneticamente. A cepa possui um gene reporter, o LacZ de Escherichia coli que sintetiza uma enzima, a À-galactose que pode catalisar uma reacao colorimetrica com o substrato vermelho de clorofenol À-galactopiranosideo (CPRG). A atividade enzimatica e diretamente proporcional ao numero de parasita. No presente trabalho propomos a introducao de uma tecnica colorimetrica para caracterizacao de parametros para quantificacao do parasitismo tecidual e comparacao com a tecnica histologica e Real Time-PCR. Camundongos Balb/C machos foram divididos em grupos controle (nao infectado) e experimental variando a via de inoculacao, a saber, intraperitoneal (IP), subcutanea (SC) e oral (OR). As curvas parasitemicas foram realizadas por meio da coleta de sangue da cauda do animal. Apos morte dos mesmos, foram estudados os tecidos cardiaco, hepatico, esplenico, enterico e muscular (musculo esqueletico). Os orgaos foram retirados, divididos em tres porcoes equitativas para analise histologica, enzimatica e molecular. Na atividade enzimatica, comparando os tecidos em suas vias de inoculacao, o figado apresentou resultado significativo no grupo inoculado via IP. Tecido cardiaco apresentou diferenca estatistica (P<0,05) entre as vias de inoculacao OR e SC, apresentando maior atividade enzimatica na primeira, o que esta relacionado a uma maior quantidade de parasita tecidual. Nos cortes histologicos o grupo SC mostrou-se mais infectado com grande numero de ninhos de forma amastigota em praticamente todos os tecidos, enquanto nos demais grupos foram observados ninhos de amastigota em sua maioria no coracao e viii musculo esqueletico. A presenca do parasita nos demais tecidos foi confirmada pela eletroforese em gel de agarose 1,5% apos tecnica de PCR convencional. A Real Time-PCR nao se mostrou satisfatoria no trabalho devido a dificuldade na sua padronizacao. Concluindo, a metodologia enzimatica se mostrou favoravel e adequada na quantificacao parasitaria tecidual, podendo ser aperfeicoada para corroborar sua eficacia. / One hundred years after the discovery of Chagas Disease, an illness caused by blood born protozoan Trypanosoma cruzi, is still consider a public health problem. It is a major cause of morbidity and mortality in Central and South Americas where more than 8 million individuals are already infected. This illness can be contracted by different forms as vector transmission, blood transfusion, tissue transplant, laboratory accidents and oral route. The parasite has a heterogeneous genotype and phenotype due to accumulating mutation in their sub-populations. CL Brener is considered as a pattern strain due to some intrinsic characteristics as low infectivity, good response to therapy drugs and a stable genetic heritage. The CL B5 clone of T. cruzi was originated by a genetic modification of the CL Brener strain which has a reporter gene LacZ of Escherichia coli, which induces the syntesis of â-galactosidase which is able to catalyze a colorimetric reaction using the substrat Chlorophenol red-â-D-galactopyranoside (CPRG). The enzymatic activity is directly related with the number of the parasites. In this study we proposed a new colorimetric assay to quantify T. cruzi load in different animal tissues by comparing with other methodologies such histological and molecular assay (Real Time-PCR). Male Balb/C mice were separated into control and experimental groups according to the route of infection (IP-intraperitoneal, SC-subcutaneous, OR-oral). Parasitemic curves were made by collecting blood samples from the tail of experimental animals. Fifteen days after, animals were euthanized and their tissues removed (heart, liver, spleen, intestine and skeletal muscles) and divided in three portions for the colorimetric analyses, histopathology and molecular assay. The enzymatic activity was performed comparing the number of parasites in different tissues according to the route of infection. Comparing them, it was observed that heart and liver displayed the highest number of parasites as compared with the other studied tissues. In the liver, IP route triggered the highest number of parasites while in the heart OR and SC routes displayed enhanced parasites. The histopathology analysis revealed that SC group presented the highest number of amastigote nests. For IP and OR groups, nests were mostly observed in heart and skeletal muscles. T. cruzi DNA were detected using electrophoreses 1,5 % agarose gel after conventional PCR technique, displaying characteristic bands of DNA. The Real Time-PCR was not a satisfactory x assay for this study due to difficulties on its patronization. To conclude, the enzymatic methodology was advantageous and appropriated on quantifying tissue parasites. Some further experiments will be needed to improve methodology and its efficacy.

Made available in DSpace on 2015-04-14T13:35:37Z (GMT). No. of bitstreams: 1 444726.pdf: 2086146 bytes, checksum: 971613dd60f265767b032ee73f8f457d (MD5) Previous issue date: 2012-10-04 / BACKGROUND: Personality traits contribute to drug related behavior, but their role on cocaine experimentation, abuse and addiction is not yet clear. Our aim was to evaluate how emotional traits and affective temperament were associated with cocaine experimentation, abuse and dependence with a recently developed scale for the assessment of emotional (e.g. Anger, Volition) and affective temperaments (e.g. cyclothymic). An anonymous web-survey provides the optimal means to evaluate sensitive issues such as drug related behavior in the general population. METHODS: the data was collected in the Brazilian Internet Study on Temperament and Psychopathology (BRAINSTEP), which included the Affective and Emotional Composite Temperament Scale (AFECTS) and the Alcohol, Smoking and Substance Involvement Screening Test (ASSIST). The final sample consisted of 28,587 subjects (26.6% males, mean age=30.8 ? 9.8 yrs). Traits analysis controlled for age, gender, ethanol and marijuana use. RESULTS: For emotional traits, Inhibition (particularly caution), Coping and Control were significantly lower in the cocaine groups compared to controls, particularly in those with cocaine dependence. Anger and Desire increased in relation to the degree of cocaine involvement. The associations with Emotional Sensitivity and Volition were less robust. For affective temperaments, increased cocaine use was related to a lower proportion of stable types (obsessive, euthymic and hyperthymic) and the anxious type, and a higher proportion of cyclothymic and euphoric temperaments in both sexes. CONCLUSION: specific externalized and unstable traits were associated with cocaine related behavior. Addressing these traits may be important for recovery and prevention strategies. / JUSTIFICATIVA: Os tra?os de personalidade contribuem para o comportamento do consumo de drogas, mas seu papel na experimenta??o, abuso e depend?ncia ? coca?na ainda n?o ? claro. Nosso objetivo foi avaliar como os tra?os emocionais e o temperamento afetivo estavam associados ? experimenta??o, ao abuso, e a depend?ncia ? coca?na com uma escala desenvolvida recentemente para a avalia??o dos tra?os emocionais (por ex: Raiva, Vontade) e dos temperamentos afetivos (por ex: ciclot?mico). Um levantamento an?nimo via web fornece o meio ideal para avaliar quest?es delicadas, tais como, o comportamento em rela??o ao consumo de drogas il?citas na popula??o em geral. M?TODOS: Os dados foram coletados atrav?s de um amplo levantamento via web chamado BRAINSTEP (Brazilian Internet Study on Temperament and Psychopathology), que inclui a Escala Composta de Temperamento Emocional e Afetivo (AFECTS) e o teste que afere o envolvimento com ?lcool, tabaco e outras subst?ncias (ASSIST). A amostra final foi de 28.587 indiv?duos (26,6% homens, com idade m?dia de 30,8 ? 9,8 anos). An?lise das vari?veis foi controlada para idade, sexo, uso de etanol e maconha. RESULTADOS: Para os tra?os emocionais, a Inibi??o (particularmente a cautela), o Coping, e o Controle foram significativamente menores nos grupos que usaram coca?na em compara??o com os controles, principalmente no grupo com depend?ncia ? coca?na. A Raiva e o Desejo estavam aumentados de acordo com o grau de envolvimento com a droga. As associa??es com a Sensibilidade e a Vontade foram menos robustas. Para o temperamento afetivo, o uso de coca?na foi relacionado a uma menor propor??o de tipos est?veis (obsessivos, eut?micos e hipert?micos) e do tipo ansioso, e uma maior propor??o dos temperamentos ciclot?micos e euf?ricos em ambos os sexos. CONCLUS?O: Tra?os espec?ficos externalizados e inst?veis foram associados com o comportamento relacionado ao uso de coca?na. O conhecimento destas caracter?sticas pode ser importante para o desenvolvimento de estrat?gias de preven??o e controle do uso desta droga.

La tuberculose (TB) est une maladie très répandue qui provoque la mort de plus d'un million de personnes dans le monde chaque année. Cela représente un véritable problème de santé publique qui nécessite le développement de nouveaux médicaments et traitements afin de mieux lutter contre cette maladie destructrice. Dans ce contexte, il est important de comprendre la relation entre l'agent étiologique, Mycobacterium tuberculosis (Mtb) et le système immunitaire. Les mycobactéries sont recouvertes de structure glycolipidiques qui pourraient être reconnus par des récepteur spécifiques appelés lectine de type C. Ici, nous avons étudié le rôle de deux lectines récemment identifiées qui n'ont pas été étudié dans un contexte de TB, CL-LK et DCIR. Au cours des dernières années, de nouvelles lectines solubles ont été identifiées et parmi elles ; CL-K1 a été définie par un criblage de divers ligands glycosylés, comme étant un potentiel récepteur de motifs présents sur la paroi de Mtb. Dans le sang, CL-K1 est associé avec une autre lectine soluble du nom de CL-L1, formant un complexe du nom de CL-LK. Par des expériences in vitro, notamment via des approches biochimique et d'analyses cytométriques, nous avons pu confirmer que CL-LK peut se lier à Mtb et encore plus particulièrement à la coiffe mannose présent sur le lipoarabinomannane, un constituant majeur de la mycomembrane. Les souris déficientes en CL-K1, une sous unités de CL-LK, ne présentent pas d'altération particulière de susceptibilité à Mtb. Cependant, nous avons pu mettre en évidence que la quantité de CL-LK dans le sérum de patient atteint d'une TB active est réduite comparé aux patients contrôles, et que cela corrèle de manière inverse à la réponse immune induite par le pathogène. Ces résultats indiquent que CL-LK pourrait présenter un intérêt comme élément de futur diagnostics ou suivi de traitement. Dans une seconde partie de la thèse, nous nous sommes concentré sur la lectine DCIR dans la réponse immune envers l'agent étiologique de la TB. DCIR est exprimé dans les lésions pulmonaires de macaques infectés par Mtb à la fois durant les infections asymptomatiques et dans les infections conduisant à une TB active. In vitro, une analyse globale de l'expression génique couplée à des validations par RT-qPCR ont pu révéler que suite à une infection par Mtb, l'expression d'un grand nombre de gènes répondant à l'interféron (IFN) de type I est inhibée dans les cellules dendritiques issues de moelle osseuses de souris déficientes en DCIR comparées à celles issues de souris sauvages. Cette inhibition corrèle avec une phosphorylation excessive de SHP2 dans les cellules dépourvues de DCIR, suivies d'une déphosphorylation de STAT1, laquelle est impliquée dans la régulation de la réponse à l'IFN de type I. L' IFN de type I est connu comme pouvant inhiber la production d' IL-12 dans les cellules dendritiques, et en effet, nous avons pu démontrer que l'inhibition de la signalisation relative à l'IFN de type I dans les cellules dendritiques déficientes en DCIR est associée avec une augmentation de la production d'IL12p70 ainsi qu'à une plus grande capacité de ces cellules à stimuler la prolifération de lymphocytes Th1 producteur d'IFN?. Les souris déficientes en DCIR contrôlent mieux l'infection que les souris sauvages, ce qui corrèle également avec une plus grande production d'IL12p70, une plus forte réponse des lymphocytes Th1, ainsi qu'à une augmentation de l'inflammation et de l'infiltration cellulaire dans les poumons des souris déficientes. Cette inflammation excessive est caractérisée par une augmentation de production du TNF-a et de iNOS dans les poumons. En conclusion, nos résultats révèlent une nouvelle voie moléculaire par laquelle les lectines peuvent moduler l'équilibre entre l'inflammation induite par l'infection et le contrôle du pathogène par l'intermédiaire de la modulation de la signalisation relative à l'IFN de type I dans les cellules dendritiques. / Tuberculosis (TB) is a wide-spread disease which causes the death of more than one million of people around the world each year. This represents a really harmfull healthcare and new drugs and treatments are always in development to better fight this destructive illness. In this context, it is crucial to understand the relation between the etiological agent, Mycobacterium tuberculosis (Mtb) and the immune system. Mycobacteria are recovered by some glycolipid structure which could be recognized by specific receptor called C-Type Lectin. Here we investigated the role of two recently highlights C-type lectins which haven't been studied in a tuberculosis context, CL-LK (Collectin Liver Kidney) and DCIR (Dendritic Cell (DC) ImmunoReceptor). In the recent years, novel soluble lectins were identified and belong these; CL-K1 (Collectin Kidney 1) was identified by a glycan array as a potential receptor in the sugar complex recognition of M.tb. In blood, CL-K1 is linked with another soluble lectin called CL-L1 (Collectin Liver 1) and form a complex CL-LK molecule. With some in vitro experiment, notably with biochemistry approach and cytometer analysis, we were able to confirm that CL-LK can bind Mtb and more particularly the mannose residue presents on the Lipoarabinomannan, a major constituent of the mycomembrane. Mice deficient in CL-K1, one of the CL-LK subunits, do not display altered susceptibility to Mtb. However, we found that the amount of CL-LK in the serum of patients with active TB is reduced, compared to that in controls, and correlates inversely to the magnitude of the immune response to the pathogen. These findings indicate that CL-LK might be of interest for future diagnostic and treatment monitoring purposes. In a second part of this thesis, we have focused our interest on the C-Type lectin DCIR in immunity to the tuberculosis agent, Mtb. DCIR is expressed in pulmonary lesions in Mtb-infected non-human primates during both symptomless infection and active TB disease. In vitro, global gene expression profiling coupled to RT-qPCR validation revealed that upon Mtb infection, the expression of a number of interferon (IFN)-responsive genes was impaired in DCIR-deficient murine bone marrow-derived DCs, compared to in wild-type cells. This inhibition correlated with an excessive phosphorylation of Src homology 2 domain tyrosine phosphatase (SHP)2 in DCIR-KO cells, followed by a subsequent dephosphorylation of Signal transducer and activator of transcription1(STAT1) which is crucially involved in the regulation of the type I IFN.Type I IFNs are known to inhibit interleukin (IL)-12 production in DCs, and indeed, we found that impaired IFN signaling in DCIR-deficient DCs was associated with an increased production of IL-12p70 and an increased ability of Mtb-infected cells to stimulate IFN?-producing Th1 lymphocyte proliferation. DCIR-deficient mice controlled Mtb infection better than wild-type animals, which correlated with an increased production of IL-12p70, an increased proliferation of Th1 lymphocytes, and an increased inflammation and cell infiltration in the lungs of DCIR-KO animals. This excessive inflammation is characterized by an increased production of tumor necrosis factor alpha (TNFa) and inducible nitric oxide synthase (iNOS) in the lungs. Taken together, our results reveal a novel pathway by which a C-Type lectin modulates the equilibrium between infection-driven inflammation and pathogen's control through sustaining type I IFN signaling in DCs.

Completando 100 anos da descoberta da molestia, a Doenca de Chagas, causada pelo agente etiologico Trypanosoma cruzi, ainda e considerada nas Americas Central e do Sul um problema de saude publica, atingindo mais de 8 milhoes de individuos. Sua transmissao ocorre de distintas maneiras, como por vetores, transfusao sanguinea, transplante de orgaos, acidentes em laboratorios, via oral. O parasita e conhecido por sua heterogeneidade no genotipo e fenotipo, baseado em mutacoes cumulativas em diferentes sub-populacoes do parasita. A cepa CL Brener e uma cepa padrao para pesquisa com diversas peculiaridades interessantes como baixa infectividade em animais, resposta aos tratamentos existentes e seus marcadores geneticos serem estaveis. O clone CL B5 de T. cruzi e originado a partir da cepa CL Brener modificada geneticamente. A cepa possui um gene reporter, o LacZ de Escherichia coli que sintetiza uma enzima, a À-galactose que pode catalisar uma reacao colorimetrica com o substrato vermelho de clorofenol À-galactopiranosideo (CPRG). A atividade enzimatica e diretamente proporcional ao numero de parasita. No presente trabalho propomos a introducao de uma tecnica colorimetrica para caracterizacao de parametros para quantificacao do parasitismo tecidual e comparacao com a tecnica histologica e Real Time-PCR. Camundongos Balb/C machos foram divididos em grupos controle (nao infectado) e experimental variando a via de inoculacao, a saber, intraperitoneal (IP), subcutanea (SC) e oral (OR). As curvas parasitemicas foram realizadas por meio da coleta de sangue da cauda do animal. Apos morte dos mesmos, foram estudados os tecidos cardiaco, hepatico, esplenico, enterico e muscular (musculo esqueletico). Os orgaos foram retirados, divididos em tres porcoes equitativas para analise histologica, enzimatica e molecular. Na atividade enzimatica, comparando os tecidos em suas vias de inoculacao, o figado apresentou resultado significativo no grupo inoculado via IP. Tecido cardiaco apresentou diferenca estatistica (P<0,05) entre as vias de inoculacao OR e SC, apresentando maior atividade enzimatica na primeira, o que esta relacionado a uma maior quantidade de parasita tecidual. Nos cortes histologicos o grupo SC mostrou-se mais infectado com grande numero de ninhos de forma amastigota em praticamente todos os tecidos, enquanto nos demais grupos foram observados ninhos de amastigota em sua maioria no coracao e viii musculo esqueletico. A presenca do parasita nos demais tecidos foi confirmada pela eletroforese em gel de agarose 1,5% apos tecnica de PCR convencional. A Real Time-PCR nao se mostrou satisfatoria no trabalho devido a dificuldade na sua padronizacao. Concluindo, a metodologia enzimatica se mostrou favoravel e adequada na quantificacao parasitaria tecidual, podendo ser aperfeicoada para corroborar sua eficacia. / One hundred years after the discovery of Chagas Disease, an illness caused by blood born protozoan Trypanosoma cruzi, is still consider a public health problem. It is a major cause of morbidity and mortality in Central and South Americas where more than 8 million individuals are already infected. This illness can be contracted by different forms as vector transmission, blood transfusion, tissue transplant, laboratory accidents and oral route. The parasite has a heterogeneous genotype and phenotype due to accumulating mutation in their sub-populations. CL Brener is considered as a pattern strain due to some intrinsic characteristics as low infectivity, good response to therapy drugs and a stable genetic heritage. The CL B5 clone of T. cruzi was originated by a genetic modification of the CL Brener strain which has a reporter gene LacZ of Escherichia coli, which induces the syntesis of â-galactosidase which is able to catalyze a colorimetric reaction using the substrat Chlorophenol red-â-D-galactopyranoside (CPRG). The enzymatic activity is directly related with the number of the parasites. In this study we proposed a new colorimetric assay to quantify T. cruzi load in different animal tissues by comparing with other methodologies such histological and molecular assay (Real Time-PCR). Male Balb/C mice were separated into control and experimental groups according to the route of infection (IP-intraperitoneal, SC-subcutaneous, OR-oral). Parasitemic curves were made by collecting blood samples from the tail of experimental animals. Fifteen days after, animals were euthanized and their tissues removed (heart, liver, spleen, intestine and skeletal muscles) and divided in three portions for the colorimetric analyses, histopathology and molecular assay. The enzymatic activity was performed comparing the number of parasites in different tissues according to the route of infection. Comparing them, it was observed that heart and liver displayed the highest number of parasites as compared with the other studied tissues. In the liver, IP route triggered the highest number of parasites while in the heart OR and SC routes displayed enhanced parasites. The histopathology analysis revealed that SC group presented the highest number of amastigote nests. For IP and OR groups, nests were mostly observed in heart and skeletal muscles. T. cruzi DNA were detected using electrophoreses 1,5 % agarose gel after conventional PCR technique, displaying characteristic bands of DNA. The Real Time-PCR was not a satisfactory x assay for this study due to difficulties on its patronization. To conclude, the enzymatic methodology was advantageous and appropriated on quantifying tissue parasites. Some further experiments will be needed to improve methodology and its efficacy.

Small and medium-sized enterprises (SMEs) in Malaysia, particularly In the information and communications technology (lCT) sector, are faced with an increasingly volatile environment. The Malaysian business scene has opened up their markets to the world where smaller businesses find themselves competing with newly launched multinational subsidiary and subdivision companies, along with the large local firms. The Malaysian Government has launched several campaigns and support for smaller local businesses to be more competitive and to continuously compete at par with these larger companies. This research project supports the Malaysian Government's objective of instilling a more structured approach towards a more competitive SME by focusing on the management of competitive information related to these companies. In recognising the rising need for competitive support, management and executives are increasingly relying on a concept called Competitive Intelligence (Cl), a systematic and ethical process for gathering, analysing, and managing information that can affect a company's plans, decisions, and operation. In managing competitive information, several companies have emerged especially to develop online tools and software that would enhance the Cl process and the value competitive intelligence brings to organisations. The success of these Cl software tools depends, however, on the sophistication of an organisation's understanding of the Cl process and scope of usage. Different companies derive different values from different approaches to competitive intelligence, and therefore require a flexible tool that is very specific to the company's needs. Therefore, this research investigated the structures and contexts of Malaysian Small and Mediumsized Enterprises (SMEs) based on competitive intelligence (Cl) concepts to derive a more customised approach to the use of Cl for SMEs in the ICT sector, as well as in the selection of appropriate Cl software. Mintzberg's approaches to analysing organisational structures and contexts, Bouthillier and Shearer's Intelligence Cycle, Herring's Key Intelligence Topics, and Davis' concept of effectiveness were used in two main stages. The first stage involved identifying the nature and range of SMEs, which exist under Malaysia's Multimedia Super Corridor, a government benchmarking body for local businesses. This gives an account, on the basis of cluster analysis, of a taxonomy of SME categories consisted of ten clusters. The relationships between the categories were also examined in the first stage of the research. The relationships and clusters found in the first part of the research offered the basis for the second part of the research, which constructs the criteria for evaluating online tools and software for competitive intelligence. The evaluation criteria are then used to evaluate eight Cl-ready software packages in finding suitable tools for the different categories of SMEs. Finally, the research concludes with a study of the prospective users' perceptions of effectiveness in SMEs drawn from the identified clusters. This 'multiple constituency' approach to understanding effectiveness evaluates both Davis' concept of effectiveness (usefulness), as well as the differential evaluations of perceived effectiveness. The research findings provide evidence of a range of SME structures in a variety of contexts. Levels of importance placed on different levels in the Cl process are identified, as well as aspects that need support, automation and/or augmentation. The software evaluation in the second part of the research provided ten recommendations of suitable software package(s) for each SME cluster. However, an initial review by SME managers of perceived effectiveness mostly did not reveal results that were parallel to the findings from the software evaluation study. All in all, the research confirms that SMEs can be analysed by clusters but further research would be necessary to confirm the effectiveness of using the recommended Cl software over a longer period of time.

Made available in DSpace on 2016-04-04T18:27:26Z (GMT). No. of bitstreams: 1 Regina Furigo.pdf: 3253353 bytes, checksum: 40a7772625492bb6a193cb7c1f5d4f54 (MD5) Previous issue date: 2002-12-11 / The purpose of the present study was to investigate the experience undergone by both the undergraduate students and the supervisor of the clinical practicum training during the fifth year of the Psychology course. The data collection was done through the writing disclosures of the students soon after a sequence of three meetings were realized with the purpose of introducing a diversity of themes to the participating students. The themes were previously determined by the researcher to function as a triggering subject. The meetings were successively carried out during the month of November, 2001 lasting approximately one and a half hours each, so that the themes could be truly discussed. The chosen themes were based not only on the relationship provided by the close contact between the supervisor and the students of the practicum clinical experience course, but also the empathic comprehension among the involved parts, in addition to and specially for the identification that the supervisor had with a crises moment that the course was going through. This situation demanded that the students articulated their experiences in relation to the graduation process in the clinical psychology arena, providing than a great opportunity to help in the process of improving the status of the course through the achievement of a renewed comprehension of its processes. In order to do this, three meetings were carried out with each one of the three-practicum experience classes under the supervision of the researcher (and clinical supervisor). All of the students of each class (approximately 15 students) agreed on participating in the study, adding to a total of forty-five students and fifteen hundred of collected disclosures. The process was done in the University setting in which the students mastered their undergraduate degree in Psychology. The fifth year students were selected to participate in this study because it was assumed that they should be in a more appropriate position to evaluate the context of the subject of study, since they already had throughoutly experienced the academic process in the psychology arena. The data was collected immediately after the end of each meeting through the disclosures of the students so that it could reflect the sense of the students experiences. The data analyzes was based on the qualitative exploration of the phenomenological orientation. The results indicated that there are meaningful themes present in the clinical formation process of the students pursuing the under-graduation degree in Psychology. Despite the fact that the students do severely criticize the graduating school, they also feel glad and trust the experience provided to them. They experienced moments of anxiety and tension during the starting period of clinical work due to the difficulty that they had in linking theory to practice. They also had difficulty to deal with the clinical system of the University, which functions as an institution that rather privilege the academic scheme instead of the professional formation of the students in order to prepare them to a competitive and demanding job-market. It was observed that during a demanding period such as the fifth year of the psychology course, the students need a structure that supports them, so they can feel accepted and better able to compensate their insecurity and anxiety. Such situations were experienced during some therapeutic circumstances that were reflected during the supervisions, due to the supervisor s capacity to form alliances with the students. In addition, there were a great number of disclosures emphasizing situations of empathy and acceptance of the students in relation to their clients worlds. However, there were also expressions of concern around the incomprehension felt by many students playing the role of a beginner therapist in relation to their clients backgrounds. This resulted in an internal conflict between their world-view and own personal experiences, and their clients . The most valuable experiences granted during the graduation process were the possibility given to the students to participate in group supervisions where they had the opportunity to exchange their own personal and academic experiences with other students and the supervisor. Finally, the mostly significant experience seems to have been the possibility of acting as a clinical psychologist for the first time. / O objetivo deste trabalho foi o de investigar a viv?ncia de forma??o cl?nica de graduandos e supervisora, de ?ltima s?rie de um curso de gradua??o. Como porta de acesso, optouse pela coleta de depoimentos escritos, logo ap?s tr?s reuni?es, cujas tem?ticas foram previamente pensadas pela pesquisadora e apresentadas ao grupo como quest?es disparadoras. As reuni?es foram consecutivas e visaram o aprofundamento do tema enfocado.Ocorreram durante o m?s de novembro de 2001, tendo a dura??o de aproximadamente uma hora e trinta minutos cada. Os temas escolhidos nasceram da conviv?ncia estreita com estagi?rios, da compreens?o emp?tica entre supervisora e supervisionando, mas, principalmente, pela identifica??o do momento de crise, no qual se inseria o curso e da necessidade dos alunos expressarem suas viv?ncias, podendo colaborar para a melhoria futura do curso, a partir de uma compreens?o renovada do mesmo. Realizaram-se ent?o, tr?s reuni?es com cada um dos tr?s grupos envolvidos, por ser este o n?mero de classes sob a responsabilidade da supervisora-pesquisadora e todos os alunos decidiram-se por participar. Cada turma tinha aproximadamente quinze alunos, num total de quarenta e cinco participantes e cento e quinze depoimentos coletados. Os encontros foram realizados nas depend?ncias da Universidade, onde os alunos conclu?am a gradua??o em Psicologia. Optou-se por alunos quintanistas por considerar-se que os mesmos, ao estarem concluindo a gradua??o, teriam condi??es de avaliar o contexto pesquisado com maior abrang?ncia, por j? terem vivido na totalidade as experi?ncias acad?micas na ?rea cl?nica. Os dados coletados atrav?s de depoimentos escritos imediatamente ap?s o t?rmino das reuni?es, por supervisionandos e supervisora, possibilitaram a apreens?o do sentido das viv?ncias, analisadas segundo pressupostos da pesquisa qualitativa explorat?ria de orienta??o fenomenol?gica. Os resultados apontaram que existem grandes temas perpassando a forma??o cl?nica dos alunos de um curso de gradua??o em Psicologia. Do mesmo modo que tecem severas cr?ticas ? agencia formadora, sentem-se gratificados e confiantes na experi?ncia que tiveram. Viveram momentos de angustia e apreens?o, ao iniciarem os seus atendimentos cl?nicos, pela dificuldade em articular ci?ncia e pr?tica. Tiveram dificuldade tamb?m em lidar com a Cl?nica-Escola enquanto Institui??o, por entenderem-na arbitr?ria, funcionando em um esquema que mais privilegia a vida acad?mica do que a pr?profissionalizac?o do aluno para um mercado de trabalho competitivo e exigente. Percebeu-se que em um ano t?o exigente, como ? o ?ltimo ano da gradua??o, o aluno precisa de estruturas institucionais que o ap?iem, por tratar-se justamente de uma Cl?nica-Escola, para que possam sentir-se um pouco mais amparados e assim compensar sua inseguran?a e ansiedade. Facilidades vividas foram apontadas em rela??o ao manejo de algumas situa??es terap?uticas, decorr?ncia direta das supervis?es. A grande habilidade sentida foi com a capacidade para forma??o de v?nculos. Foram freq?entes os relatos de situa??es de empatia e aceita??o em rela??o ao mundo do cliente, embora tamb?m tenham sido expressas dificuldades decorrentes de uma incompreens?o do referencial subjetivo de clientes, levando o terapeuta iniciante a um confronto entre o seu referencial individual e o do cliente. Os grandes momentos vividos ao longo da gradua??o localizam-se na possibilidade de participa??o em grupos de supervis?o envolvendo trocas de experi?ncias e compartilhamento de aspectos de vida pessoal e acad?mica com os parceiros de grupo e supervisor. A experi?ncia mais significativa parece ter ocorrido diante da possibilidade de atua??o como psic?logos cl?nicos.

Made available in DSpace on 2017-07-24T19:38:04Z (GMT). No. of bitstreams: 1 Juliane Nadal Dias.pdf: 2277080 bytes, checksum: cfb6cfa9f705c82f0634bb16f025fd4f (MD5) Previous issue date: 2010-09-03 / Fundação Araucária de Apoio ao Desenvolvimento Científico e Tecnológico do Paraná / Photodynamic therapy (PDT) is a kind of treatment minimally invasive which depends on two components: photosensitizer and light. It consists of the administration of a drug which after reaching its action site is activated by light, whose wavelength is absorbed and catalyses the production of free radicals oxygen, and singlet oxygen, which trigger the cellular death of the unhealthy tissue, either by necrosis or apoptosis. As this technique appears as very promising in the treatment of bacteria infections and cancers (mainly skin cancer of the non-melanoma), several studies seek new photosensitizers compounds which are efficient and present little collateral effect. The aim of this study was to investigate the photodynamic efficiency of [5, 10, 15, 20-tetrakis(4-sulfonatophenyl) porphyrinato Iron (III)], chloride, [Fe(TPPS4)]Cl, and 5, 10, 15, 20-tetrakis(N-methyl-4-pyridyl) porphyrin p-toluenesulfonate, [H2(TMPyP)], compounds, through spectroscopic and cytotoxic analysis, using LED irradiation system, for later use in photodynamic therapy. The uric acid method was employed in order to determine the photodynamic activity, analyse the spectroscopic behavior regarding alterations in the environment pH, with and without CTAB and interpret photobleaching reactions. Toxicity was evaluated through in vitro experiments with microorganisms (Escherichia coli, Staphylococcus aureus e Candida albicans) and Artêmia salina. Results presented in this work with [H2(TMPyP)] porphyrin, using LED irradiation, demonstrated that the porphyrin:LED system can be used in PDT, once it presented satisfactory photodynamic activity, and significant inhibition of CFUs (Colony forming units), regarding the micro organisms under study, mainly when the red LED was used. Metalporphyrin [Fe(TPPS4)]Cl presented lower photodynamic activity values in both AU and microorganisms tests under the experimental conditions employed when compared to [H2(TMPyP)]. / A Terapia Fotodinâmica (TFD) é uma modalidade de tratamento minimamente invasiva que depende de dois componentes: fotossensibilizador (FS) e luz. Consiste na administração de um fármaco que, ao atingir o seu sítio de ação, é irradiado por luz, cujo comprimento de onda é absorvido, e catalisa a produção de radicais de oxigênio, e de oxigênio singlete, que desencadeiam a morte celular do tecido atingido, por necrose ou apoptose. Por ser promissora no tratamento em infecções bacterianas e em cânceres (principalmente em câncer de pele do tipo não melanoma), muitos estudos buscam novos compostos fotossenssibilizadores que sejam eficazes e apresentem poucos efeitos colaterais. O objetivo desse trabalho foi investigar a eficiência fotodinâmica dos compostos cloreto de [5, 10, 15, 20-tetrakis(4-sulfonatofenil)porfirinato ferro(III)], [Fe(TPPS4)]Cl, e p–toluenosulfonato de 5, 10, 15, 20-tetrakis(N-metil-4-piridil)porfirina, [H2(TMPyP)], através de ensaios espectroscópicos e citotóxicos utilizando sistema de LED para irradiação, visando posterior utilização em Terapia Fotodinâmica. Utilizou-se o método do ácido úrico para determinação da atividade fotodinâmica, a análise do comportamento espectroscópico a partir de alterações do pH do meio, na ausência e presença de CTAB, interpretações de reações de fotobranqueamento, e avaliou-se a atividade fotodinâmica através de ensaios in vitro com micro-organismos (Escherichia coli, Staphylococcus aureus e Candida albicans) e Artêmia salina. Os resultados apresentados neste trabalho com a porfirina [H2(TMPyP)] utilizando-se LED na irradiação, demonstraram que o sistema porfirina/LED tem potencial para ser utilizado em TFD, uma vez que apresentou atividade fotodinâmica satisfatória, e uma inibição significativa nas UFCs dos micro-organismos estudados, principalmente quando se fez uso de LED vermelho. A metaloporfirina [Fe(TPPS4)]Cl apresentou menores valores de atividade fotodinâmica no teste de AU e nos testes com micro-organismos, nas condições experimentais utilizadas, quando comparada com [H2(TMPyP)].

Las membranas celulares contienen un canal de Cl- (Maxi-Cl-) que es modulado por estrógenos y antiestrógenos. El trabajo experimental de mi tesis doctoral ha consistido en el estudio de la modulación de este canal y su identidad molecular. El resultado de mi trabajo ha demostrado que la base molecular del canal Maxi-Cl- es una isoforma de la proteína mitocondrial VDAC y que la activación del canal por los antiestrógenos y su inhibición por los estrógenos implica procesos de defosforilación y fosforilación, respectivamente. / Cellular membranes contain a Maxi-Cl- channel that is modulated by oestrogen and antioestrogens. The experimental work of my PhD Thesis has focussed in the study of the molecular identity of the Maxi-Cl- channel and its regulation. My results demonstrated that an isoform of the mitochondrial protein VDAC is the molecular correlate of the Maxi-Cl- channel and that the activation of the channel by antioestrogens and its inhibition by oestrogen requires a dephosphorylation and phosphorylation process, respectively.

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