Submitted by Mariane Bertolucci Casalenovo null (mary_bc14@hotmail.com) on 2017-09-14T00:20:08Z No. of bitstreams: 1 DISSERTAÇÃO FINAL.pdf: 10749096 bytes, checksum: 4412c078d6e18d64f519d7ac0952d26d (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-09-15T13:49:22Z (GMT) No. of bitstreams: 1 casalenovo_mb_me_bot.pdf: 10749096 bytes, checksum: 4412c078d6e18d64f519d7ac0952d26d (MD5) / Made available in DSpace on 2017-09-15T13:49:22Z (GMT). No. of bitstreams: 1 casalenovo_mb_me_bot.pdf: 10749096 bytes, checksum: 4412c078d6e18d64f519d7ac0952d26d (MD5) Previous issue date: 2017-08-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A infecção por Mycobacterium leprae (M. leprae) desencadeia alterações no status funcional das células de Schwann (SCs), responsáveis pela produção da bainha de mielina e homeostasia neural. Estudos prévios indicam que lesões nos nervos periféricos modulam a expressão de fatores-chave envolvidos na diferenciação e maturação das SCs. Na hanseníase, alterações funcionais nessas células podem estar envolvidas na patogênese do dano neural. O presente estudo buscou determinar a expressão dos fatores de transcrição KROX-20, SOX-10, JUN e do receptor p75NTR, envolvidos nos processos de diferenciação e maturação de SCs. Os dados foram obtidos a partir de cultura primária de SCs murinas expostas ao M. leprae durante sete e 14 dias, com diferentes multiplicidades de infecção (MOI 100:1, MOI 50:1). Os mesmos fatores foram avaliados in vivo, em nervos ciáticos de camundongos nude (NUFoxn1nu), após seis e nove meses de infecção. Comparando-se grupos experimentais em relação aos controles, nossos resultados em cultura de SCs demonstram redução significativa de KROX-20 e SOX-10, bem como elevação na frequência de células imunomarcadas por p75NTR. Paralelamente, nervos ciáticos de camundongos nude infectados por M. leprae apresentaram queda de KROX-20 e aumento de p75NTR, quando comparados aos animais não infectados. Em conjunto, nossos resultados corroboram achados prévios sobre a interferência de M. leprae na expressão de fatores-chave envolvidos na maturação celular, favorecendo a manutenção de fenótipo não mielinizante nas SCs, com possíveis implicações na reparação de nervos periféricos adultos. / Mycobacterium leprae infection (M. leprae) triggers changes in the functional status of Schwann cells (SCs), which are responsible for the production of myelin sheath and neural homeostasis. Previous studies indicate that peripheral nerve injury modulates the expression of key factors involved in the differentiation and maturation of SCs. In leprosy, functional changes in these cells may be involved in the pathogenesis of neural damage. The present study sought to determine the expression of the transcription factors KROX-20, SOX-10, JUN and the neurotrophic receptor p75NTR, involved in the differentiation and maturation processes of SCs. Data were obtained from primary murine SCs culture exposed to M. leprae for seven and 14 days, with different multiplicities of infection (MOI 100:1, MOI 50:1). The same factors were evaluated in vivo on sciatic nerve of nude mice (NU-Foxn1nu), after six and nine months of infection. By comparing experimental groups versus controls, our SC culture results demonstrate significant reduction of KROX-20 and SOX-10, as well as elevation in the frequency of cells immunolabelled by p75NTR. In parallel, sciatic nerves of nude mice infected with M. leprae showed a decrease in KROX-20 and an increase of p75NTR when compared to uninfected animals. Our results corroborate previous findings on M. leprae interference in the expression of key factors involved in cell maturation, favoring the maintenance of a non-myelinating phenotype in SCs, with possible implications for the repair of adult peripheral nerves.

Thesis (Master Diploma (Medical Technology) -- Cape Technikon, Cape Town, 1992 / The steady state in a tumour rapidly changes with its growth and the subsequent deteriorating blood and nutrient supply. This adaptation in the steady state of the tumour is shown in the increased lactate dehydrogenase and acid phosphatase activity in the tumour during it's growth. These alterations in the tumour metabolism places an increased burden on the body to supply nutrient and to discard the waste products of the tumour. This is demonstrated at the macroscopic level by the decreasing body weight and food intake when the tumour burden increases, and also at the metabolic levels by the responses of certain glycolytic and Cori cycle enzymes. Furthermore three distinct stages were observed in the Corl cycle response to the influence of the tumour namely, a silent or preclinical stage, a hypermetabolic stage and a hypometabolic stage. Although the decreasing body weight cannot be directly linked to the process of gluconeogenesis, the onset of anorexia appeared to coincide with the end of the hypermetabolic stage and the beginning of the hypometabolic stage in gluconeogenesis. This clearly shows that the body's steady state is adversely affected by the presence of the tumour and that the conditions at the metabolic level seem to cause the anorexia. Furthermore, it is well known that the success of cancer therapies depends entirely on the effectiveness o{the modality to kill the tumour cell and on the ability . of the host to absorb the damage caused by the modality without being destroyed in the process itself. The second part of this study demonstrates the radioprotective effects of ATP at all levels. It is clear from this work that ATP had a bigger influence in protecting the normal tissue than it had on the tumour tissue. This was demonstrated by the response of acid phosphatase (AP) and glucose-6-phosphate dehydrogenase (G-6-PDH) in the tumour and testis. Furthermore, it would seem that ATP has a multifactorial interaction with the cell, two possible mechanisms of protection are indicated by these results. The fIrst of these interactions is through the receptors of the cell to stimulate enhanced glycolysis, for higher energy production and thus repair. The second possibility is the interaction of ATP with the receptor of the cell to inhibit the production of free radicals and thus damage, as demonstrated by the response of G-6-PDH and AP.

Thesis (BTech (Biomedical Technology))--Cape Technikon, 1992. / The steady state in a tumour rapidly changes with its growth and the subsequent deteriorating blood and nutrient supply. This adaptation in the steady state of the tumour is shown in the increased lactate dehydrogenase and acid phosphatase activity in the tumour during it's growth. These alterations in the tumour metabolism places an increased burden on the body to supply nutrient and to discard the waste products of the tumour. This is demonstrated at the macroscopic level by the decreasing body weight and food intake when the tumour burden increases, and also at the metabolic levels by the responses of certain glycolytic and Cori cycle enzymes. Furthermore three distinct stages were observed in the Cori cycle response to the influence of the tumour namely, a silent or preclinical stage, a hypermetabolic stage and a hypo metabolic stage. Although the decreasing body weight cannot be directly linked to the process of gluconeogenesis, the onset of anorexia appeared to coincide with the end of the hypermetabolic stage and the beginning of the hypometabolic stage in gluconeogenesis. This clearly shows that the body's steady state is adversely affected by the presence of the tumour and that the conditions at the metabolic level seem to cause the anorexia. Furthermore, it is well known that the success of cancer therapies depends entirely on the effectiveness ofthe modality to kill the tumour cell and on the ability' of the host to absorb the damage caused by the modality without being destroyed in the process itself. The second part of this study demonstrates the radioprotective effects of ATP at all levels. It is clear from this work that ATP had a bigger influence in protecting the normal tissue than it had on the tumour tissue. This was demonstrated by the response of acid phosphatase (AP) and glucose-6-phosphate dehydrogenase (G-6-PDH) in the tumour and testis. Furthermore, it would seem that ATP has a multifactorial interaction with the cell, two possible mechanisms of protection are indicated by these results. The first of these interactions is through the receptors of the cell to stimulate enhanced glycolysis, for higher energy production and thus repair. The second possibility is the interaction of ATP with the receptor of the cell to inhibit the production of free radicals and thus damage, as demonstrated by the response of G-6-PDH and AP.

Thesis (MTech (Medical Technology))--Cape Technikon, 1992. / The steady state in a tumour rapidly changes with its growth and the subsequent deteriorating blood and nutrient supply. This adaptation in the steady state of the tumour is shown in the increased lactate dehydrogenase and acid phosphatase activity in the tumour during it's growth. These alterations in the tumour metabolism places an increased burden on the body to supply nutrient and to discard the waste products of the tumour. This is demonstrated at the macroscopic level by the decreasing body weight and food intake when the tumour burden increases, and also at the metabolic levels by the responses of certain glycolytic and Cori cycle enzymes. Furthermore three distinct stages were observed in the Cori cycle response to the influence of the tumour namely, a silent or preclinical stage, a hypermetabolic stage and a hypometabolic stage. Although the decreasing body weight cannot be directly linked to the process of gluconeogenesis, the onset of anorexia appeared to coincide with the end of the hypermetabolic stage and the beginning of the hypometabolic stage in gluconeogenesis. This clearly shows that the body's steady state is adversely affected by the presence of the tumour and that the conditions at the metabolic level seem to cause the anorexia. Furthermore, it is well known that the success of cancer therapies depends entirely on the effectiveness of the modality to kill the tumour cell and on the ability' of the host to absorb the damage caused by the modality without being destroyed in the process itself. The second part of this study demonstrates the radioprotective effects of ATP at all levels. It is clear from this work that ATP had a bigger influence in protecting the normal tissue than it had on the tumour tissue. This was demonstrated by the response of acid phosphatase (AP) and glucose-ó-phosphate dehydrogenase (G-6-PDH) in the tumour and testis. Furthermore, it would seem that ATP has a multifactorial interaction with the cell, two possible mechanisms of protection are indicated by these results. The first of these interactions is through the receptors of the cell to stimulate enhanced glycolysis, for higher energy production and thus repair. The second possibility is the interaction of ATP with the receptor of the cell to inhibit the production of free radicals and thus damage, as demonstrated by the response of G-6-PDH and AP.

Since before the opening of the University of British Columbia medical school in 1950, a group of Vancouver citizens has contested the use of laboratory animals by local scientists. The resulting debate has consistently centered around questions of the cruelty and scientific value of animal experimentation. Although antivivisectionists received little coverage in Vancouver's decidedly pro-vivisectionist mainstream press between 1950 and 1980, they nevertheless caused Vancouver researchers to employ a number of tactics to foster a positive image of their animal care practices during this period. By the early 1980s, Vancouver antivivisectionists had succeeded in disseminating highly graphic descriptions of animals undergoing experimentation via local community newspapers, and in using direct action tactics to link these images with specific Vancouver laboratories. In response, medical researchers heightened their longstanding efforts to conceal their experimental practices from public view. The limited public visibility of the animal lab and the commonly held belief in the necessity of animal use for medical progress both helped to limit opposition to animal experimentation between 1950 and 1990, despite an increasingly widespread acknowledgement of the cruelty of this set of practices. / Arts, Faculty of / History, Department of / Graduate

Neurogenesis is an important and vastly under-explored area in reptiles. While the ability to generate new brain cells in the adult mammalian brain is limited, reptiles are able to regenerate large populations of neuronal cells. Pythons exhibit a characteristic specific dynamic action (SDA) response after food intake with an increase in metabolic rate that facilitates processing the meal. Associated with this change in SDA, pythons (Python spp.) also exhibit impressive plasticity in their digestive and cardiovascular physiology due to the sheer magnitude of the increase in organ growth that occurs after a meal to speed digestion, absorption, and assimilation of nutrients. While this systemic growth in response following food consumption is well documented, whether the python brain exhibits associated changes in cell proliferation following food consumption and digestion is currently unexplored. For this study, juvenile male ball pythons (Python regius) were used to test the hypothesis that postprandial neurogenesis is associated with food consumption. We used the thymidine analog 5-bromo-12’-deoxyuridine (BrdU) to quantify and compare cell proliferation in the brain of fasted snakes and at two time points: two days and six days after a meal, which span time periods of during and after SDA response, respectively. Quantification of BrdU-labeled cells in the ventricular regions relealed that – consistent with other reptile species – the retrobulbar and olfactory regions had the highest numbers of proliferating cells in the python brain, regardless of sampling time. Throughout the telencephalon, cell proliferation was significantly greater in the six-day post-feeding group, with no difference between the two-day post-feeding group and controls. Most other postprandial systemic plasticity occurs within a day or two after a meal and decreases thereafter; however, the brain displays a more delayed response, with a surge of cell proliferation after most of the digestion and absorption is complete. Our results support our hypothesis that food consumption does affect cell proliferation in the python brain, and indicates that the degree of increased proliferation is dependent on the time since feeding.

Investigations of human biology and disease have been hindered by the use of animal models. The information obtained from such studies often results in clinically irrelevant results and drug trial failures. Additionally, several governing bodies have been formulating legislation to move away from animal models and toward more ethical and efficient testing platforms for drug discovery and cosmetic research. As an answer to these issues, "body-on-a-chip" systems have been a rapidly developing field which easily recapitulates in vivo functionality, providing a more relevant, repeatable, and ethical testing platform to better predict biology. These systems can be used as human-based testing platforms to evaluate human physiology, disease progression, and drug responsiveness for specific cell types and multi-organ systems. Diseases such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) have significant research challenges, specifically with translating research findings into treatment plans. The complexity of the neuromuscular reflex arc, the biological system affected by these diseases, is difficult to study with traditional molecular techniques, namely because the many components of this disease system interact with each other using complex pathways. This work pushes the existing platform to a more complete human model of neuromuscular disease with the incorporation of gamma motoneurons, development of the first human induced pluripotent cell (iPSC) derived intrafusal fibers, and proposals to incorporate nociceptive neurons all on a functionally interrogative platform. The incorporation of these components will allow for a more complete, clinically relevant model to study neuromuscular disorders and for preclinical dug discovery.

Across species, the face and more specifically the mouth, serves as an essential facet of everyday life. Amongst humans the mouth serves as a tool for the ingestion of food, a marker for facial recognition and a medium for communication. In order for the mouth to properly form, a series of precise growth and fusion events are needed. In order to insure that these events are orchestrated properly is a wide array of signals, transcription factors and epigenetic regulators. Due to the needed precision of these events, congenital birth defects of the face such as cleft lip and cleft palate are some of the most common worldwide. In order to support existing and identify new developmental processes involved in mouth formation, we have utilized the effective model, Danio to study the molecules and events implicated in orofacial development. This was accomplished by developing a novel confocal imaging technique that allows for visualization of the forward facing zebrafish. Using this imaging technique we were able to establish when the embryonic mouth first forms in zebrafish. Additionally, we recapitulated cleft-palate phenotypes shown in previous literature with the imaging method. Utilizing this technique, we then sought to further establish the role of Ca2+ signaling in proper orofacial morphogenesis and determine if the serine/threonine protein kinase, Ca2+/calmodulin-dependent protein kinase type-II (CaMK-II), has a role in proper orofacial developmental.

O presente trabalho propõe-se a analisar a controvérsia da experimentação animal por meio do método da dialética materialista a partir de duas unidades que compõem a temática: o teor dos argumentos usados por diferentes atores na controvérsia e as contradições que engendram os sistemas de atividade dos sujeitos. Por meio da dialética, buscou-se superar a dualidade dos argumentos apresentados, na busca por novas formas de divulgação científica. Sem julgar a validade e o mérito das argumentações, foram analisados o discurso de duas pesquisadoras, dois ativistas, um político e um representante do Conselho Nacional de Controle de Experimentação Animal. A partir do arcabouço estrutural da Teoria da Atividade, proposto por Engeström, buscamos compreender onde se localizam as contradições mais evidentes no discurso dos sujeitos e discutir como ações de divulgação científica poderiam propiciar um ambiente favorável para a superação dessas contradições e o desenvolvimento qualitativo do sistema de atividade. Por essa razão, queremos trazer, para esta investigação, as conferências de consenso como ferramenta inovadora de comunicação da ciência, em uma abordagem deliberativa e com participação ativa da sociedade, que nos auxilie no avanço desses conflitos. / This study proposes to analyze the controversy of animal experiments by the method of materialist dialectics from two units that make up the theme: the content of the arguments used by different actors in the controversy and contradictions that engender the activity systems subject. Through the dialectic, he sought to overcome the duality of the arguments made in the search for new forms of science communication. Without judging the validity and the merits of the arguments, we analyzed the speech of two researchers, two activists, a politician and a representative of the National Council for Animal Experimentation Control. From the structural framework of Activity Theory, proposed by Engeström, we try to understand where there are the most obvious contradictions in the discourse of subjects and discuss how science communication actions could provide a favorable environment for overcoming these contradictions and the qualitative development of the system activity. For this reason, we want to bring to this research, consensus conferences as an innovative communication tool of science, in a deliberative approach and active participation of society, to assist in the advancement of these conflicts.

Thesis (M.A.)--University of North Carolina at Wilmington, 2003. / Includes bibliographical references (leaves : [84]-89).