Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Submitted by Rafael Vieira (vieira.rafa1986@gmail.com) on 2016-01-06T18:57:51Z No. of bitstreams: 1 dissertacao final rafael.pdf: 11211535 bytes, checksum: e982b9dd12cb7bf43a451db3f5e30076 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-01-06T19:24:55Z (GMT) No. of bitstreams: 1 vieira_r_me_araiq_par.pdf: 1125273 bytes, checksum: 201f41df3c870e0118ce7d37d6eca396 (MD5) / Made available in DSpace on 2016-01-06T19:24:55Z (GMT). No. of bitstreams: 1 vieira_r_me_araiq_par.pdf: 1125273 bytes, checksum: 201f41df3c870e0118ce7d37d6eca396 (MD5) Previous issue date: 2015-12-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Este trabalho de mestrado tem, como foco principal, analisar o comportamento do fungo Xylaria sp. perante variações físico-químicas, através da abordagem OSMAC (One Strain, Many Compounds). Para realizar tais induções e explorar a variabilidade da rede metabólica deste micro-organismo, um planejamento fatorial foi desenhado visando induzir a variabilidade (ou a potencialização) de metabólitos. Diante de respaldos quimiométricos, induções planejadas foram impostas ao micro-organismo e observou-se variações no perfil químico nos extratos brutos. Somado a isso, técnicas de desreplicação foram usadas a fim de estudar quimicamente as matrizes complexas produzidas por este fungo visando a produção de metabólitos de alto valor agregado e determinou-se condições específicas para produção de moléculas de citocalasina D e de griseofulvina. / This work has as its main focus, analyze the behavior of the fungus Xylaria sp. before physicochemical variations via the OSMAC approach (One Strain, Compounds Many). To achieve such induction and exploit the variability of the metabolic network of this micro-organism, a factorial design was designed to induce variability (or potentiating) metabolites. Before chemometric backrests, planned inductions were imposed on the micro-organism and it was observed variations in the chemical profile in crude extracts. Added to this, dereplication techniques were used to study the chemically complex matrices produced by this fungus aimed at producing high added value and metabolites was determined specific conditions for the production of molecules cytochalasin D and griseofulvin.

As dermatofitoses dos carnívoros domésticos são infecções fúngicas superficiais, causadas habitualmente por dois gêneros fúngicos: Microsporum sp e Trichophyton sp. Trata-se de uma antropozoonose, com importância na saúde pública Objetivou-se: caracterizar a população de cães e gatos acometidos por dermatofitose atendidos no Serviço de Dermatologia do HOVET /USP num período de 27 meses; determinar a valia do exame histopatológico como metodologia diagnóstica; e comparar a eficácia da griseofulvina e da terbinafina na terapia das dermatofitoses. Foram atendidos 76 animais com diagnóstico de dermatofitose, 47,3% pertenciam à espécie fetina e 52,7% à canina O agente etiológico isolado com maior freqüência em caninos e felinos foi Microsporum canis. Não se observou distribuição sazonal. Os cães de raça definida foram os mais acometidos (75,0%), dentre estes, aqueles da raça Yorkshire Temer (23,3%). Os felinos com e sem definição racial igualmente acometidos, porém os Persas (93,7%) foram os mais acometidos dentre aqueles com plena definição. Observou-se que a maioria dos animais infectados apresentava menos de um ano de idade (65,8%). As lesões mais observadas foram: alopecia, eritema, escamas e crostas. A maioria das lesões apresentava configuração circular e estavam localizadas nas regiões cefálicas, de tronco e de membros. O prurido esteve ausente em 50,0% dos caninos e em 88,8% dos felinos. As lesões dos cães tinham caráter mais inflamatório. O exame histopatológico mostrou-se pouco sensível (28,6%) e com alto número de falsos negativos (71,4%). A griseofulvina (5Omg/kg/dia) foi eficaz em l00,0% dos casos, sem acarretar efeitos colaterais, com média de tempo para cura de 41 dias. Já a terbinafina na dose de cinco mg/kg/dia, apresentou eficácia de 81,8%, sem induzir efeitos colaterais e com êxito terapêutico em 21 dias. Demonstrou-se assim, que a dose de 20 mg/kg/dia demonstrou a mesma eficácia que a dose de cinco, porém, com efeitos colaterais em 16,6% dos animais tratados, com tempo médio para cura de 33 dias. Demonstrou-se que a terbinafina é uma boa alternativa terapêutica, porém, a griseofulvina ainda se constitui na droga de eleição para o tratamento das dermatofitoses de caninos e felinos. / Dermatophytosis in domestic carnivorous are superficial infections caused mainly by two genus of fungus: Microsporum sp and Trichophyton sp. This disease is an anthropozoonosis important for public health. The goals of this study were: characterize the population of cats and dogs with dermatophytosis treated in the Dermatology Service of HOVET FMVZ/USP in a period of 27 months; evaluate the validity of the histopathological exam as a methodology of diagnosis; and compare the efficacy of griseofulvin and terbinafine in the therapy of the dermatophytosis. Seventy six animals (47,3% were felines and 52,7% were canines) were evaluated in this study. The more frequent isolated etiological agent in canines and felines was Microsporum canis. Seasonality was not observed. The dogs with a defined breed were more predisposed (75,0%) and the Yorkshire Terrier dogs had a higher proportion of positive cultures (23,3%). The felines with or without breed definition got the same frequency, however, the Persians (93,7%) were more predisposed among those of pure breed. It was noticed that the majority of infected animals were under one year of age (65,8%). The most observed lesions were: alopecia, crusts, erythema and scales. The majority of the lesions had a circular form and were found in the cephalic, trunk and limb regions. There was no pruritus in 50,0% of the canines and 88,8% of the felines. The lesions were more inflammatory in dogs. The histopathological exam had a low sensitivity (28,6%) and a high number of false negatives (71,4%).Griseofulvin (50 mg/kg/day) was effective in 100,0% of the cases, with no side effects, the average time for cure was 41 days. The terbinafine used at the dosage of 5 mg/kg/day showed an efficacy of 81,3%, no side effects were observed as well and average time for cure was 21 days. The same efficacy using the dosage of 20 mg/kg/day of terbinafine was observed, although side effects were observed in 16,6% of the animals treated with an average time for cure of 33 days. The present study demonstrated that terbinafine is a good therapeutical alternative, although griseofulvin is still the first choice drug for the treatment of dermatophytosis in dogs and cats.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / The low solubility of griseofulvin in water (1.4 and 1.95 mg/dL at 25 and 37 ÂC, respectively) is a limiting factor for its administration. Despite of this, micellar solutions prepared with copolymers are being used to increase the solubility of these drugs. In addition, these mixtures of surfactants have low toxicity, are efficient in the dilution process and present high potential as carriers of insoluble drugs. These factors are the main reason for the interest in the research of these copolymers. This project proposes the investigation of the use of copolymers P123 (E21P67E21), E45S8 (denominated S8) and E45S17 (denominated S17) and their mixtures to obtain higher solubility of the drug griseofulvin through the method of direct dilution. The project also aims to investigate of the potential of these systems as a drug carrier model for griseofulvin through the study of cytotoxicity of these carriers through human neutrophil, the study of the gelation properties of the two diblocks S8 and S17 and its mixtures with triblock P123 in the proportion of 50/50 and 30/70. To determine the critical micelle concentration (cmc) the method of dye solubilisation was used to measure the fluorescence. The solubility of the drug griseofulvin in the micellar systems was quantified by UV-Vis spectrophotometer and by 1H NMR. The systems with pure S17 and with encapsulated griseofulvin were characterised by infrared spectroscopy and through x-ray diffraction. Experiments of drug release in vitro were carried out with the best encapsulating systems or carriers. The copolymers S8 and S17 and their mixtures with the P123 showed to be promising for the administration of hydrophobic drugs as they presented low toxicity and excellent values for solubilisation capacity (Scp). The mixtures P50/S8 and P50/S17 at 37 oC were the ones that presented the best results of Scp with equally low values of cmc, therefore showing the efficiency of these micellar systems as carriers of the drug griseofulvin. / A baixa solubilidade da griseofulvina em meio aquoso (1,4 e 1,95 mg/dL a 25 e 37 ÂC, respectivamente) Ã fator limitante para sua administraÃÃo. No entanto, soluÃÃes micelares preparadas com copolÃmeros vÃm sendo utilizadas para aumentar a solubilidade desses fÃrmacos. AlÃm disso, essas misturas de copolÃmeros apresentam baixa toxicidade, eficiÃncia no processo de dissoluÃÃo como tambÃm apresentam elevado potencial como nanocarreadores de fÃrmacos poucos solÃveis, sendo estes fatores determinantes para despertar interesse nas pesquisas com copolÃmeros. Esse trabalho propÃe investigar o uso dos copolÃmeros P123 (E21P67E21), E45S8 (denominado S8) e E45S17 (denominado S17) e suas misturas para serem utilizados como carreadores do fÃrmaco modelo griseofulvina, desenvolvendo estudos de liberaÃÃo, solubilizaÃÃo e citotoxicidade em neutrÃfilo humano, com o objetivo de se obter capacidades mais elevadas de solubilizaÃÃo do fÃrmaco griseofulvina. Utilizou-se a tÃcnica de inversÃo de tubo para o estudo das propriedades geleificantes dos diblocos S8 e S17 e suas misturas com o tribloco P123 nas proporÃÃes 50/50 e 30/70. A mistura P50/S8 foi a que melhor conservou as propriedades termorresponsivas do P123. Os sistemas com o S17 puro e com a griseofulvina encapsulada foram caracterizados por tÃcnicas de espectroscopia de absorÃÃo na regiÃo do infravermelho (FT-IR) e por difraÃÃo de raios-X. Os resultados de FT-IR nÃo apresentaram mudanÃas significativas para S17 e ele encapsulado com a griseofulvina. Entretanto, os resultados de raios-x confirmaram que o fÃrmaco encontra-se encapsulado no sistema polimÃrico na forma amorfa. Experimentos de liberaÃÃo in vitro foram realizados para os melhores sistemas encapsulantes ou carreadores. A mistura P50/S17 foi a que apresentou maior porcentagem de liberaÃÃo do fÃrmaco que foi de 48%. A citotoxicidade foi investigada atravÃs da atividade da LDH em neutrÃfilo humano (5 x 106 cÃlulas/mL). Em geral, os copolÃmeros nÃo mostraram citotoxicidade. A solubilidade do fÃrmaco griseofulvina nos sistemas micelares foi quantificada por espectrofotometria UV-Vis e por RMN de 1H. Para a determinaÃÃo da concentraÃÃo micelar crÃtica (cmc) utilizou-se o mÃtodo de solubilizaÃÃo de corante medida por fluorescÃncia. As misturas P50/S8 e P50/S17 a 37 ÂC foram as que apresentaram melhores resultados de Scp como tambÃm baixos valores de cmc, mostrando, portanto, a eficiÃncia desses sistemas micelares como carreadores do fÃrmaco griseofulvina.

Indiana University-Purdue University Indianapolis (IUPUI) / Prevention of pathological angiogenesis is a key strategy for treatment of common blinding ocular diseases such as retinopathy of prematurity, proliferative diabetic retinopathy, and wet age-related macular degeneration. The current treatment strategies are associated with partial vision loss and are ineffective in a significant patient population. Hence novel drugs as well as new ways to target ocular angiogenesis are needed for treating these diseases. I pursued a natural antiangiogenic compound, cremastranone, to develop novel drug leads and to find new targets. The objective of my doctoral thesis project was to elucidate cremastranone’s molecular mechanism of action and optimize its structureactivity relationship (SAR). In order to achieve this goal, with the help of chemistry collaborators cremastranone was synthesized for the first time. I showed that cremastranone has 50-fold more potency against endothelial cells as compared to nonendothelial cells, and also tested a novel active isomer, SH-11052. By SAR studies I identified a potent molecule, SH-11037, that has 10-fold more selectivity against retinal endothelial cells as compared to macrovascular endothelial cells. I then elucidated cremastranone’s molecular mechanism using a chemical proteomic approach. I identified ferrochelatase (FECH) as a specific interacting protein partner of cremastranone using photoaffinity chromatography. Hence, I hypothesized that cremastranone exerts its antiangiogenic activities through modulation of the functions of FECH. Cremastranone inhibited the enzymatic activity FECH in endothelial cells. Therefore, I investigated the role of FECH in ocular angiogenesis. Partial loss of FECH, using a siRNA-based knock down approach, decreased retinal angiogenesis both in vitro and in vivo in mouse models. Knock down of FECH decreased the expression levels of key proangiogenic proteins HIF-1α, eNOS, and VEGFR2. This work suggests that ferrochelatase plays an important, previously undocumented role in angiogenesis and that targeting of this enzyme by cremastranone might be exploited to inhibit pathological angiogenesis in ocular diseases.