Submitted by Priscila dos Santos Oliveira (priscilaspitty@hotmail.com) on 2018-08-29T22:43:22Z No. of bitstreams: 1 Merged_Priscila Oliveira.pdf: 1205779 bytes, checksum: 2ed8db0dfdefdc0a97d7a31fad8acaa9 (MD5) / Approved for entry into archive by Silvana Alvarez null (silvana@ict.unesp.br) on 2018-09-03T14:47:25Z (GMT) No. of bitstreams: 1 Oliveira_mp_me_sjc.pdf: 1205779 bytes, checksum: 2ed8db0dfdefdc0a97d7a31fad8acaa9 (MD5) / Made available in DSpace on 2018-09-03T14:47:25Z (GMT). No. of bitstreams: 1 Oliveira_mp_me_sjc.pdf: 1205779 bytes, checksum: 2ed8db0dfdefdc0a97d7a31fad8acaa9 (MD5) Previous issue date: 2018-08-06 / Resumo: A epidemiologia das infecções causadas por Candida sofreu mudanças nos últimos anos. Embora Candida albicans ainda seja o principal patógeno causador de candidíase, relatos recentes reportam o aumento da incidência de infecções causadas por espécies de Candida não-albicans. A toxicidade dos fármacos antifúngicos utilizados juntamente com o desenvolvimento da resistência tem se tornado um sério problema clínico. Com isso, atualmente existe a urgência na descoberta de novos compostos com atividades antifúngicas. Objetivos: avaliar a atividade antifúngica do fármaco Cloridrato de Verapamil, determinar a Concentração Inibitória Mínima (CIM) e a Concentração Fungicida Mínima (CFM), frente à Candida albicans ATCC 18804, Candida krusei ATCC 6258, Candida parapsilosis ATCC 90018 e Candida glabrata ATCC 9030. Além de avaliar a citotoxicidade do fármaco em queratinócitos humanos (HaCaT). Metodologia: a determinação da CIM foi realizada pela técnica de microdiluição de acordo com The European Committee on Antimicrobial Susceptibility Testing (EUCAST). A CFM foi determinada por plaqueamento em ágar Sabouraud de alíquotas provenientes das diluições do ensaio de microdiluição. A avaliação da citotoxicidade foi realizada pela técnica de redução da resazurina, sendo possível avaliar a atividade mitocondrial das células HaCaT. Resultados: o fármaco Cloridrato de Verapamil demonstrou atividade antifúngica contra as quatro espécies patogênicas de Candida, com o valor de CIM de 1250 µM; valor de CFM maior que 1250 µM, sendo assim este fármaco foi considerado fungistático. Além disso, o Cloridrato de Verapamil não apresentou citotoxicidade nas concentrações avaliadas no estudo, pois, a redução de células viáveis nas concentrações mais elevadas não ultrapassa 30%. Com relação à redução de biomassa em biofilme de Candida, após tratamento com Cloridrato de Verapamil, houve redução (de 10 a 20%) para as quatro espécies de Candida em estudo, quando utilizadas concentrações de fármaco correspondentes a CIM; quando utilizadas concentrações de fármaco correspondentes a cinco vezes o valor de CIM, houve um aumento significativo na redução de biomassa (de 25% a 60%) em biofilme formado pelas quatro espécies de Candida. Conclusão: o fármaco Cloridrato de Verapamil apresentou atividade antifúngica para Candida albicans e Candida não-albicans, sendo considerado um fármaco fungistático, além de não apresentar citotoxicidade em queratinócitos humanos, e demonstrar atividade na redução de biomassa em biofilme formado pelas quatro espécies de Candida. Demais estudos são necessários para verificar a ação desse fármaco em diferentes mecanismos de virulência frente a células de Candida spp. / Abstract: The epidemiology of Candida infections has changed in recent years. Although Candida albicans is still the main pathogen causing candidiasis, recent reports have reported an increase in the incidence of infections caused by nonalbicans Candida species. The toxicity of the antifungal drugs used together with the development of resistance has become a serious clinical problem. With this, there is now an urgency in the discovery of new compounds with antifungal activities. Objectives: To evaluate the antifungal activity of the drug Verapamil Hydrochloride, to determine the Minimum Inhibitory Concentration (MIC) and the Minimum Fungicidal Concentration (CFM) against Candida albicans ATCC 18804, Candida krusei ATCC 6258, Candida parapsilosis ATCC 90018 and Candida glabrata ATCC 9030 In addition to evaluating the cytotoxicity of the drug in human keratinocytes (HaCaT). Methodology: MIC determination was performed by the microdilution technique according to The European Committee on Antimicrobial Susceptibility Testing (EUCAST). The CFM was determined by plating on Sabouraud agar from aliquots from the dilutions of the microdilution assay. The evaluation of cytotoxicity was performed using the resazurin reduction technique, and it was possible to evaluate the mitochondrial activity of HaCaT cells. Results: The drug Verapamil Hydrochloride demonstrated antifungal activity against the four pathogenic species of Candida, with MIC value of 1250 μM; CFM value greater than 1250 μM, so this drug was considered fungistatic. In addition, Verapamil Hydrochloride did not present cytotoxicity in the units evaluated in the study, because the reduction of viable cells in the most cells is not exceeded by 30%. Regarding the biomass reduction in Candida biofilm, after treatment with Verapamil Hydrochloride, there was a reduction (from 10 to 20%) for the four Candida species in study, when the use of drug corresponding to MIC; when the use of drug corresponding to 5 times of MIC, there was a significant increase in the biomass reduction (from 25% to 60%) in the biofilm molded by the four Candida species. Conclusion: Thus, the drug Verapamil Hydrochloride led to the antifungal activity of Candida albicans and non-albicans Candida species, being considered a fungicidal drug, besides not presenting cytotoxicity in human serotonizers, and were submitted to biomass reduction in biofilm molded by four Candida species. More studies are needed to verify the action of the drug on different mechanisms of virulence against Candida spp cells.

This diploma thesis aims to optimalization the process of magnetron sputtering and creating of thin layers for use in advanced oxidation processes. During the work was created range of TiOx layers. For this process was used physical method of sputtering called PVD. The photocatalytic activity of the deposited films was tested by degradation of organic dyes Acid Orange 7. Furthermore, the layer was analyzed on surface morphology (SEM) and the layer thickness (profilometry). Study of created layers was focused on the link between the characteristics of each layer, deposition parameters and photocatalysis properties. Based on these results, the layers were applied in a system using AOP for the decomposition of organic substances. The first chapter is devoted to a summary of existing knowledge of photocatalysis and its principles. Another chapter is devoted to the theory and methods of applying thin layers and summary of knowledge of the low-pressure discharges. In the exprimental section there are described various components of the apparatus. Furthermore, the experimental part of the work focuses on the analysis of the optimization process of applying thin layers on titanium oxide. The last chapter of the thesis contains the results of the experiments on the basis of is designed another research progress of this issue.