Submitted by Geandra Rodrigues (geandrar@gmail.com) on 2018-06-19T12:42:02Z No. of bitstreams: 1 lorenaevangelistafernandes.pdf: 3413787 bytes, checksum: 3ec0f338e3d4a5a74fb584252625026d (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2018-06-27T11:14:53Z (GMT) No. of bitstreams: 1 lorenaevangelistafernandes.pdf: 3413787 bytes, checksum: 3ec0f338e3d4a5a74fb584252625026d (MD5) / Made available in DSpace on 2018-06-27T11:14:53Z (GMT). No. of bitstreams: 1 lorenaevangelistafernandes.pdf: 3413787 bytes, checksum: 3ec0f338e3d4a5a74fb584252625026d (MD5) Previous issue date: 2018-02-28 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / A tradição e a qualidade na produção de queijos no estado de Minas Gerais é destaque no cenário nacional. A Microrregião do Campo das Vertentes foi reconhecida oficialmente como região produtora do queijo Minas artesanal pelo Instituto Mineiro de Agropecuária (IMA) em 2009 e conta com um grande número produtores familiares que preservam a produção em pequena escala com a utilização de leite cru e métodos tradicionais. No Brasil, queijos elaborados a partir de leite cru possuem exigências legislativas que controlam seu comércio nas esferas municipais, estaduais e federais. Sendo assim, o presente trabalho teve como objetivo criar uma alternativa aos laticínios para produzir um queijo similar aqueles encontrados no mercado local e regional a partir de leite pasteurizado e culturas lácteas industriais, podendo ser comercializado em qualquer região do país sob inspeção. Para isso, após a identificação da unidade industrial (laticínio) e queijaria tradicional, localizados na Microrregião do Campo das Vertentes, foi elaborado um plano de fabricação, maturação, coleta e transporte dos queijos junto aos produtores. Foram feitas análises do perfil de textura (TPA), de aspectos físico-químicos, microbiológicas e sensoriais dos queijos artesanais tradicionais e dos queijos produzidos a partir da tecnologia desenvolvida ao longo de 60 dias de maturação. Adotou-se o delineamento em parcelas subdivididas no tempo e os resultados obtidos foram submetidas à análise estatística ANOVA e a Teste de Tukey em nível de 5% de significância por meio do software SISVAR. O emprego de leite pasteurizado e de culturas lácteas mistas não influenciou no perfil de textura dos queijos industriais, avaliados aos 2, 20, 40 e 60 dias após a fabricação, em relação aos artesanais (p>0,05). Da mesma forma, o perfil microbiológico, analisado aos 2 e 60 dias de maturação apresentou contagens de coliformes a 30 ºC, coliformes a 45 ºC, Staphylococcus aureus, Listeria spp. e Salmonella spp. abaixo dos limites estabelecidos pela legislação vigente, não apresentando diferenças entre tratamentos. A análise de aspectos físico-químicos, feita aos 2, 20, 40 e 60 dias, mostrou que a tecnologia não influenciou os teores de cloreto de sódio, umidade e resíduo mineral fixo dos queijos (p>0,05). Atividade de água, gordura, pH, proteína total, extensão e profundidade de proteólise apresentaram diferenças significativas em períodos pontuais entre os queijos artesanais e industriais (p<0,05). Dos atributos sensoriais avaliados pela equipe técnica de pesquisa da EPAMIG ILCT, textura, gosto salgado e odor não diferiram estatisticamente entre tratamentos (p>0,05). Já o aspecto global dos queijos foi melhor avaliado nos queijos artesanais aos 40 dias quando comparados aos industriais no mesmo período. A consistência, o aroma e o sabor obtiveram notas mais altas ao fim da maturação (60 dias) e a cor interna e sabor residual obtiveram melhor avaliação a partir dos 20 dias nos queijos artesanais. Nos testes de intenção de compra (FACT) e aceitação (Escala Hedônica) não foi observada influência do fator tratamento (p>0,05). A tecnologia desenvolvida para fabricação de QMA com emprego de leite pasteurizado e cultivos lácteos industriais é uma alternativa possível para produção em laticínios, podendo gerar benefícios sociais e econômicos como forma de auxílio na consolidação do estado de Minas Gerais como principal e mais tradicional produtor de queijos do Brasil, seja em nível artesanal como industrial. / Cheese-making tradition and quality in the state of Minas Gerais has stood out in the national scenario. The Campo das Vertentes region was officially recognized as a Minas cheese producing region by the Minas Gerais Institute of Agriculture (IMA) in 2009, with on-farm small-scale cheese making using raw milk and traditional methods. In Brazil, cheeses made from raw milk have legislative requirements that regulate marketing in the municipal, state, and federal spheres. Thus, the present study aimed to study an alternative to the dairy industry to produce cheese similar to those found in the local market, using pasteurized milk and industrial cultures, thus allowing product commercialization throughout the country under inspection. For that, after the selection of the dairy processing unit and the traditional cheese factory, located in the Campos das Vertentes region, a plan of manufacture, ripening, collection, and transport of cheeses was established together with the producers. The traditional artisanal cheese and industrial cheese produced using the technology under study were characterized for the texture profile (TPA), and physicochemical, microbiological, and sensory evaluation during 60 days of ripening. A split plot design was used, and the results were submitted to ANOVA and Tukey's test at a 5% level of significance using the SISVAR software. The use of pasteurized milk and mixed starter cultures did not affect the texture profile of the industrial cheeses after 2, 20, 40, and 60 days of ripening, when compared to the artisanal cheeses (p>0.05). Concerning the microbiological profile after 2 and 60 days of ripening, counts of coliforms at 30ºC, coliforms at 45ºC, Staphylococcus aureus, Listeria spp., and Salmonella spp. were below the limits established by the current legislation, with no significant differences between the treatments. With regard to the physicochemical characteristics, sodium chloride, moisture levels, and fixed mineral residue of cheeses were not affected after 2, 20, 40, and 60 days of ripening (p>0.05). In contrast, significant differences (p<0.05) were observed for water activity, fat, pH, total protein, extent and depth of proteolysis between the artisanal and industrial cheeses during storage. The sensory evaluation performed by the EPAMIG ILCT research team showed no significant differences for texture, salty taste, and odor between treatments (p>0.05). On the other hand, higher overall impression scores were obtained for the artisanal cheeses when compared to the industrial cheeses at 40 days of ripening. In addition, higher consistency, aroma, and flavor scores were observed at the end of ripening (60 days), while the attributes internal color and residual flavor of the artisanal cheeses exhibited higher scores after 20 days of ripening. The treatment did not affect the purchase intention (FACT) and acceptance test (hedonic scale) (p>0.05). The technology developed to make artisanal Minas cheese using pasteurized milk and industrial cultures can be an effective alternative for the dairy industry, and can generate social and economic benefits, allowing the consolidation of the state of Minas Gerais as the leading cheese producer at both the artisanal and industrial levels in Brazil.

A terapia fotodinâmica (TFD) é uma modalidade clínica para tratar uma variedade de neoplasias, doenças de pele e representa um promissor tratamento estético. O presente trabalho avaliou os aspectos histopatológicos e moleculares do tratamento pela TFD mediada por azul de metileno (TFD-AM) no modelo experimental in vivo do Carcinoma Espinocelular Cutâneo (CEC) e na pele sadia de camundongos Swiss. O protocolo da TFD foi de uma única sessão, com aplicação da solução de AM a 1% seguido por irradiação com laser diodo na dose total de 24 J/cm2 nos tecidos tumorais e sadios. Os animais foram sacrificados em dois períodos, 24 horas e 15 dias após TFD. Alterações morfológicas foram pouco marcantes nos tecidos tumorais tratados, entretanto, foram mais pronunciadas nos tecidos sadios. Podemos concluir que os efeitos de uma única sessão da TFD mediada pelo AM na dose aplicada não conferiu melhora no tratamento do CEC. Estes resultados motivam novos estudos com ajustes no protocolo para melhorar a eficácia desta terapia. / Photodynamic therapy (PDT) is a clinical method for treating a variety of tumors, skin disorders and represents a promising cosmetic treatment. This study evaluated the histopathological and molecular aspects of the treatment by PDT mediated by methylene blue (PDT-MB) in vivo experimental model of cutaneous squamous cell carcinoma (SCC) and in healthy skin of swiss mice. The PDT protocol was a single session with the application of MB 1% solution followed by irradiation with diode laser at a total dose of 24 J/cm2 in tumor and healthy tissue. The animals were sacrificed at two periods, 24 hours and 15 days after PDT. Morphological changes were less marked in the tumor tissues treated, however, were more pronounced in healthy tissues treated. We can conclude that the effects of a single session of PDT mediated by MB in applied dose conferred no improvement in the treatment of SCC. These results motivate further studies with adjustments in the protocol to improve the effectiveness of this therapy.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Soursop (Annona muricata L.) is a tree of Annonaceae family widely distributed in all tropical regions of the world, produces fruit known as graviola and its leaves are used in traditional medicine as phytotherapy. Some compounds present in A. muricata have been related to antioxidant activity, anti-inflammatory and antimicrobial activities of this plant. This study aimed to evaluate the antioxidant, anti-inflammatory and antibacterial activities of hexane (HE) and ethanol (EE) soursop leaves extracts. To this end, we used soursop leaves for preparation of extracts for further analyzes of phytochemical composition, antioxidant capacity, ear edema assays in vivo and in vitro antibacterial activity. For analysis of the antioxidant activity of hexane and ethanol extracts were determined total polyphenols, yellows flavonoids and anthocyanins and total antioxidant capacity by ABTS&#9679;+ and DPPH methods. To evaluate the anti-inflammatory activity of the extracts was performed ear edema model induced by xylene or TPA in mice (Swiss females, 25-35g)-treated intragastrically (ig) for 7 consecutive days or topically (single dose) of different concentrations of the extracts (EE or HE):10, 100 and 1000 mg/kg. The antimicrobial activity of the extracts was evaluated against strains of Staphylococcus aureus (ATCC 14458), Listeria monocytogenes (ATCC 33090), Escherichia coli (ATCC 11775) and Salmonella choleraesuis (ATCC 14028). The extracts showed wide variety of phytochemical compounds, especially EE, and high antioxidant activity. Pretreatment via i.g. with the extracts was able to inhibit the edema induced by xylene, with a maximum percentage of inhibition of 61.72% for EE 10 and 57.08% for HE 100, but no effect on TPA-induced edema. The topical application with HE inhibited in a dose-dependent form the xylene-induced edema with maximum inhibition of 78.89% for HE1000, while for EE1000 was 67.95%, with no significant difference EE10 and EE100. As for the TPA-induced edema, HE (10, 100 and 1000 mg / kg) inhibited edema at 84.39%, 68.68% and 65.96%, while for EE (10, 100 and 1000 mg / kg ) the inhibition was 75%, 56% and 51%, respectively. Moreover, the extracts showed no antibacterial activity against all tested strains. The hexane and ethanol extracts of soursop leaves exhibit high differentiated antioxidant and anti-inflammatory activity compared to edema induced by xylene and TPA. / A gravioleira (Annona muricata L.) à uma Ãrvore da famÃlia Annonaceae largamente distribuÃda em todas as regiÃes tropicais do mundo, produz fruto conhecido como graviola e suas folhas sÃo usadas na medicina tradicional como fitoterÃpicos. Alguns compostos presentes na A. muricata jà foram relacionados com as atividades antioxidante, anti-inflamatÃria e antimicrobiana dessa planta. Esse trabalho teve por objetivos avaliar as atividades antioxidante, anti-inflamatÃria e antibacteriana dos extratos hexÃnico (EH) e etanÃlico (EE) das folhas de gravioleira. Para tanto, foram utilizadas folhas de gravioleira para o preparo dos extratos utilizados em anÃlises da composiÃÃo fitoquÃmica, da capacidade antioxidante total, ensaios de edema de orelha in vivo e da atividade antibacteriana in vitro. Para a anÃlise da atividade antioxidante dos extratos hexÃnico e etanÃlico foram determinados os polifenÃis totais, flavonoides amarelos e antocianinas totais e a capacidade antioxidante total pelos mÃtodos ABTS&#9679;+ e DPPH. Para avaliaÃÃo da atividade anti-inflamatÃria dos extratos, foi utilizado o modelo de edema de orelha induzido por xileno ou TPA em camundongos (Swiss fÃmeas, 25-35g) submetidos a tratamentos por via intragÃstrica (i.g.) por 7 dias consecutivos ou por via tÃpica (dose Ãnica) com diferentes concentraÃÃes dos extratos (EH ou EE): 10, 100 e 1000 mg/kg. A atividade antimicrobiana dos extratos foi avaliada contra cepas de Sthaphylococcus aureus (ATCC 14458), Listeria monocytogenes (ATCC 33090), Escherichia coli (ATCC 11775) e Salmonella choleraesuis (ATCC 14028). Os extratos apresentaram grande variedade de compostos fitoquÃmicos, principalmente o EE, alÃm de elevada atividade antioxidante. O prÃ-tratamento por via i.g. com os extratos foi capaz de inibir o edema induzido por xileno, com um percentual mÃximo de inibiÃÃo de 61,72% para o EE10 e de 57,08% para o EH100, mas nenhum efeito no edema induzido por TPA. O tratamento tÃpico por EH inibiu de forma dose-dependente o edema induzido por xileno, com mÃximo de inibiÃÃo de 78,89% para EH1000, enquanto para EE1000 foi de 67,95%, sem diferenÃa significativa com EE10 e EE100. Jà para o edema induzido por TPA, EH (10, 100 e 1000 mg/kg) inibiram o edema em 84,39%, 68,68% e 65,96%, enquanto para EE (10, 100 e 1000 mg/kg) a inibiÃÃo foi de 75%, 56% e 51%, respectivamente. Ademais, os extratos nÃo apresentaram atividade antibacteriana frente Ãs cepas testadas. Os extratos hexÃnico e etanÃlico das folhas de gravioleira apresentam elevada capacidade antioxidante e atividade anti-inflamatÃria diferenciada em relaÃÃo aos edemas induzidos por xileno e TPA.

A terapia fotodinâmica (TFD) é uma modalidade clínica para tratar uma variedade de neoplasias, doenças de pele e representa um promissor tratamento estético. O presente trabalho avaliou os aspectos histopatológicos e moleculares do tratamento pela TFD mediada por azul de metileno (TFD-AM) no modelo experimental in vivo do Carcinoma Espinocelular Cutâneo (CEC) e na pele sadia de camundongos Swiss. O protocolo da TFD foi de uma única sessão, com aplicação da solução de AM a 1% seguido por irradiação com laser diodo na dose total de 24 J/cm2 nos tecidos tumorais e sadios. Os animais foram sacrificados em dois períodos, 24 horas e 15 dias após TFD. Alterações morfológicas foram pouco marcantes nos tecidos tumorais tratados, entretanto, foram mais pronunciadas nos tecidos sadios. Podemos concluir que os efeitos de uma única sessão da TFD mediada pelo AM na dose aplicada não conferiu melhora no tratamento do CEC. Estes resultados motivam novos estudos com ajustes no protocolo para melhorar a eficácia desta terapia. / Photodynamic therapy (PDT) is a clinical method for treating a variety of tumors, skin disorders and represents a promising cosmetic treatment. This study evaluated the histopathological and molecular aspects of the treatment by PDT mediated by methylene blue (PDT-MB) in vivo experimental model of cutaneous squamous cell carcinoma (SCC) and in healthy skin of swiss mice. The PDT protocol was a single session with the application of MB 1% solution followed by irradiation with diode laser at a total dose of 24 J/cm2 in tumor and healthy tissue. The animals were sacrificed at two periods, 24 hours and 15 days after PDT. Morphological changes were less marked in the tumor tissues treated, however, were more pronounced in healthy tissues treated. We can conclude that the effects of a single session of PDT mediated by MB in applied dose conferred no improvement in the treatment of SCC. These results motivate further studies with adjustments in the protocol to improve the effectiveness of this therapy.

A Thesis submitted to The University of Arizona College of Medicine - Phoenix in partial fulfillment of the requirements for the Degree of Doctor of Medicine. / Current stroke imaging protocol at Barrow Neurological Institute calls for a noncontrast computed tomography (NCCT), a computed tomography angiography (CTA), and a computed tomography perfusion (CTP) at the time of presentation to the emergency department (ED), and follow up imaging includes magnetic resonance diffusion weighted imaging (MR‐DWI). This information is used to determine the appropriateness and safety of tissue plasminogen activator (tPA) administration. Previous studies have shown the risk for post‐tPA hemorrhagic conversion rises significantly as the size of the infarct core increases. Thus, it is of great importance to have an accurate method of measuring core infarct size in patients presenting with acute ischemic stroke. The purpose of our study is to determine if CTP correctly identifies the infarct core and if post‐tPA hemorrhagic conversion is related to the size of the infarct core and/or the accuracy of CTP in identifying the infarct core. The ultimate goal is to improve patient outcomes by decreasing the morbidity and mortality associated with tPA administration. This study is a retrospective chart review of all patients who presented to the ED during a one year period with signs and symptoms of acute ischemic stroke who then subsequently received tPA. Imaging was also reviewed, including the NCCT, CTA, CTP, and MRDWI for each patient. In this study, MR‐DWI is used as the gold standard for determining the presence or absence of an infarct core. CTP and MR‐DWI are in agreement of the presence of an infarct core in 7 patients, or 10 percent of the time. Similarly, CTP and MR‐DWI are in agreement of the absence of an infarct core in 31 patients, or 44 percent of the time. In the other 32 patients, CTP and MR‐DWI are in disagreement. The percent correlation between CTP and MR‐DWI was found to be 24 percent with a p‐value < 0.05. As for post‐tPA hemorrhagic conversion, 12 percent of patients had hemorrhagic conversion, and when the hemorrhage rate was compared to the size of the infarct core, the odds of post‐tPA hemorrhagic conversion were 56 times higher in the group of patients with infarct cores larger than one‐third of a vascular territory than in patients with smaller infarct cores with a p‐value < 0.001. Although no significant correlation was found between the accuracy of CTP data and the rate of post‐tPA hemorrhagic conversion, patients with concordant CTP and MR data had a 46% lower likelihood of post‐tPA hemorrhagic conversion than did patients with contradictory CTP and MR‐DWI data. Conclusion: Because patients with infarct cores larger than one‐third of a vascular territory are 56 times more likely to hemorrhage than patients with smaller infarct cores and CTP is less accurate than MR‐DWI in identifying the infarct core in patients presenting with acute ischemic stroke, CTP studies should not be part of the acute stroke imaging protocol. Another imaging modality, such as MR‐DWI, may be preferential in the setting of acute ischemic stroke to identify the infarct core.

Cette thèse présente l’étude des effets des radiations sur des composants analogique du commerce. Il est mis en place une méthode de caractérisation expérimentale de la sensibilité des composants électroniques par le biais de simulation électrique et de stimulations laser. Grâce à la simulation électrique, nous classons par sensibilité des structures académiques, composant un amplificateur opérationnel, face à la propagation d’un signal transitoires. Le laser impulsionnel a permis d’extraire la sensibilité des composants testés en fonction de leur topologie. La technique d’absorption deux photons est utilisée afin d’extraire en trois dimensions le volume sensible d’un transistor / In space, natural radiation induces dysfunction in electronic device. Particle strikes silicon and generate transient current. We study this single event transient in fast commercial amplifiers. We investigate sensitivity of devices with simulation and laser property. SPICE simulation allows us to investigate the propagation of SET inside classical structure like current mirror or push pull. This work was applied at each stage of the amplification chain of an amplifier. In the other part we use laser bench with non linear property, due to non-linear absorption in silicon, to extract the sensitive volume

Universidade Federal Rural da Amazônia / In the Amazon In the Amazon there are areas of soil affected by prehistoric man, these areas are characterized by dark coloring, remains of archaeological material (ceramic fragments and lithic artifacts). Such soils are known as Archaeological Black Earth (TPA), Terra Preta de Indio or Terra Preta. These soils generally have high fertility, high levels of P, Ca, Mg, Zn, Mn, and high content of stable organic matter besides very different physical properties of the soils that originated them. The present work aims to analyze the morphological properties and to detail the behavior of the physical-water properties of areas with different concentrations of ceramic fragments as well as an area without anthropic interference of the Archaeological Site of São José located on the banks of the Xingu River in the low Xingu region , Para. For this purpose, deformed and undisturbed samples were collected from four different areas of an archaeological site in order to evaluate the behavior of the main soil properties in the different areas of occupation and concentration of archaeological material. Soil density (Ds), particle density, soil water retention curve (CRAS), total porosity, penetration resistance (PR) and stability of aggregates via water percolation were determined. Aiming at this, to increase the knowledge of Terra Preta Archaeological sites in the Amazon region in order to subsidize works of management and recovery of degraded soils. The results showed that there was dominance of the sand fraction in the studied areas, the soils being classified with a sandy loam texture. The areas with higher concentrations of ceramic materials and presence of anthropic horizon showed a marked physical quality in relation to total porosity, percolation stability and available structural water content when compared to the respective adjacent soil. Keywords: Archaeological Black Earth. Ceramic fragments. Anthropogenic soils. / Na Amazônia existem áreas de solos afetadas consideravelmente pelo homem pré-histórico, estas áreas são caracterizadas pela coloração escura, restos de material arqueológico (fragmentos cerâmicos e de artefatos líticos). Tais solos são conhecidos como Terra Preta Arqueológica (TPA), Terra Preta de Índio ou Terra Preta. Estes solos geralmente apresentam alta fertilidade, elevados teores de P, Ca, Mg, Zn, Mn, e teor elevado de matéria orgânica estável além de propriedades físicas bem distintas dos solos que as originaram. O presente trabalho tem como objetivo analisar as propriedades morfológicas e detalhar o comportamento das propriedades físico-hídrica de áreas com diferentes concentrações de fragmentos cerâmicos assim como uma área sem interferência antrópica do Sitio arqueológico São José localizado as margens do Rio Xingu na região do baixo Xingu, Pará. Para isso foram coletadas amostras deformadas e indeformadas de quatro áreas distintas de um sitio arqueológico afim de se avaliar o comportamento das principais propriedades do solo nas diferentes áreas de ocupação e concentração de material arqueológico do mesmo. Determinou-se, granulometria, densidade do solo (Ds), densidade da partícula, curva de retenção de água no solo (CRAS), porosidade total, Resistencia a penetração (RP) e estabilidade de agregados via percolação de água. Visando assim, ampliar os conhecimentos de sítios Arqueológicos de Terra Preta na região amazônica a fim de subsidiar trabalhos de manejo e recuperação de solos degradados. Os resultados demonstraram que houve a dominância da fração areia nas áreas estudadas sendo os solos classificados com textura franco argilo arenosa. As áreas com maiores concentrações de materiais cerâmicos e presença de horizonte antrópico apresentaram uma acentuada qualidade física com relação a porosidade total, estabilidade de percolação e conteúdo de água disponível estrutural quando comparada ao respectivo solo adjacente.

Malignant glioma consists of a group of diseases where the localisation and the nature of the disease makes treatment an extreme challenge. Two important biological features of malignant glioma cells are their infiltrative growth and their ability to induce angiogenesis. Glioma cells migrate extensively behind the blood-brain barrier and infiltrate the surrounding brain making radical treatment with surgery and radiotherapy almost impossible. The aims of this thesis were to investigate factors of importance for glioma cell migration and angiogenesis and to evaluate if an anti-angiogenesis approach alone or in combination with current treatment modalities could inhibit glioma growth. For this purpose we used the BT4C orthotopic rat glioma model and investigated treatment effects of the vascular endothelial growth factor (VEGF) receptor-2 and epidermal growth factor (EGF) receptor tyrosine kinase inhibitor ZD6474 alone or in combination with temozolomide or radiotherapy. Altered protein expression pattern after anti-angiogenesis treatment was measured using a mass-spectrometric proteomic method, followed by multivariate data-analysis. The tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1), and VEGF showed altered temporal and spatial mRNA expression during glioma progression. In early stages of tumour progression the expression was found throughout the tumour while in later stages, the expression was more predominant in the invasive tumour border. ZD6474 in monotherapy significantly inhibited tumour growth in the BT4C glioma model. The effect was further enhanced when combined with radiotherapy or temozolomide. Using mass-spectrometric methods an altered protein expression pattern after ZD6474 treatment was observed implicating the possibility to use proteomic methods for finding predictive biomarkers for anti-angiogenesis treatment. In conclusion, this thesis demonstrates a co-expression of factors important for glioma growth and angiogenesis and that treatment with an angiogenesis inhibitor has additive effects on glioma growth when combined with radiotherapy and chemotherapy. Finally, an altered protein expression pattern after anti-angiogenesis treatment is evident and detectable. Hopefully this work will contribute to and encourage further research to reach a better understanding of how to combine and evaluate different treatment approaches in malignant glioma.

The healing of tympanic membrane (TM) perforations is a complex wound healing process including inflammation, migration of keratinocytes and tissue remodelling. Most TM perforations in human heal spontaneously, however some perforations become chronic, and the reason to why is still largely unknown. In cutaneous wound healing plasminogen (plg) has been shown to play an important role. Plg is converted into the protease plasmin regulated by two plasminogen activators (PA), urokinase type PA (uPA) and tissue-type PA (tPA). The aim of the present thesis was to evaluate the role of plg in healing of TM perforations, both in vivo and in vitro. The main objectives were to determine the healing capacity of the TM, the involvement of keratinocytes, fibrin(ogen) and inflammatory cells in the healing process. The studies were performed in plg deficient and uPA deficient mice, with littermate wild type (wt) mice as controls It was shown that myringotomies of the TMs in plg deficient mice still remained open 143 days following a perforation. The wound area was characterized by an abundant recruitment and accumulation of inflammatory cells; mainly macrophages and neutrophils, an arrested keratinocyte migration and a fibrin deposition covering the surface of the TM. The TM perforations in the wt mice all healed within 11 days. Interestingly, the myringotomies of the plg deficient mice could be closed by reconstitution with systemic injections of plg, whereas injections of PBS had no affect on the healing. To characterize mechanisms involved in the development of persistent TM perforations in plg deficient mice after a myringotomy the early inflammatory response during the first 48 hours was studied. The recruitment and accumulation of inflammatory cells in the perforated TMs was found to be similar between the plg deficient and the wt mice. Myringotomized TMs in uPA deficient mice healed similar to perforations of wt controls. Neither did the keratinocyte migration nor the occurrence of inflammatory cells differ between these genotypes. In the in vitro experiments TMs from plg deficient and wt mice, were dissected out, perforated and cultured in absence or surplus of plg. A decrease in perforation size was seen in all groups regardless of genotype or amount of plg in the medium. In conclusion, the present studies show: • Plg is essential for the healing of TM perforations in mice. • The altered healing process after a myringotomy in plg deficient mice involves a disturbed keratinocyte migration, a massive deposition of fibrin and an abundant accumulation of inflammatory cells in the wound area. • Plasminogen deficiency does not alter the early inflammatory response, following a myringotomy. • Deficiency of uPA does not influence the healing of TM perforations. • During in vitro conditions healing of TM perforations is initiated irrespectively of genotype of the explant (plg deficient or wt) or supply of plg. The increased knowledge of the involvement of plg in the healing of TM perforations may open therapeutical possibilities in the treatment of chronic TM perforations in humans.

Ce travail de thèse a contribué au développement de la techniques de test par faisceau laser en mode d'absorption deux photons (TPA). Cette technique a plus spécifiquement été utilisée pour la détection de défauts et l'injection de fautes dans les circuits intégrés.