Made available in DSpace on 2014-10-09T12:41:53Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:09:30Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Dissertação (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:11/05408-2

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Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2015-12-10T17:52:55Z No. of bitstreams: 0 / Made available in DSpace on 2015-12-10T17:52:55Z (GMT). No. of bitstreams: 0 / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Made available in DSpace on 2015-04-01T12:09:00Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1738258 bytes, checksum: 9ea7722b27270b8d6f33deb6238bb982 (MD5) Previous issue date: 2010-12-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Streptococcus mutans is the main etiologic agent of dental caries, especially due to its ability to adhesion to the tooth surface. This bacterium produces glycosyltransferases that synthesize polymers of soluble and insoluble glucan from sucrose, which increases the colonization of cariogenic bacteria and promote the formation of biofilm on the surface of the teeth. Chlorhexidine is the most frequent topical antibiotic used in dentistry and is considered standard in the various dental specialties, but there are few studies on this drug at the molecular level. The aim of the present study was to investigate the antibacterial activity of chlorhexidine gluconate against in vitro planktonic and biofilm Streptococcus mutans cells in a dose- and timedependent manner. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of chlorhexidine in planktonic cells and MBC in biofilms were determined by microdilution method. Total S. mutans RNA from either planktonic cells or biofilms exposed or non-exposed (controls) to chlorhexidine were extracted, purified and reversely transcribed to cDNA. Real-time reverse transcription- PCR was used to quantify the relative levels of 16S rRNA, gtfB, gtfC and gtfD transcription of S. mutans in the presence of CHX. The activity of CHX in the initial biofilm structure for 2 and 4 h and morphological alterations in planktonic cells, under a range of CHX concentrations, was examined by Scanning Electron Microscopy (SEM). CLX MIC and MBC for planktonic cells were 2.2 mg/L and 18 mg/L, respectively, while MBC for biofilm was 800 mg/L. Planktonic cells exposed to CLX 4.5 mg/L and 9 mg/L were reduced almost by 6- and 20-fold, whereas biofilm counts were reduced (2.5-fold or more) in concentrations above 500 mg/L. In planktonic cells, exposition to CHX 4.5 mg/L increased gtfC and gtfD expression by 11-fold and 4-fold (p<0.01), respectively. In biofilm, expression of gtfB, gtfC and gtfD were reduced (<1.6-fold p<0.01) at concentrations above 500mg/L. Cell surfaces did not show any change when planktonic cells were exposed to CHX at 4.5 mg/L for 2 or 4h, but after 6 h, several wilted S. mutans cells with lost intracellular material could be observed. A decrease in the cells chain length and matrix was found when the initial biofilm was exposed from 1.1 mg/L to 4.5 mg/L of CHX, while 1200 mg/L and 2000 mg/L caused extensive precipitation of unknown material on the slide. Thus, we conclude that the CHX effects against bacteria depend on the kind of growth organization and the concentration/time of exposure to the drug. CHX may affect cell walls and intervene with mechanisms of the biofilm formation, and at sub-lethal concentrations, affect the expression of gtfs, which may exert an anticariogenic effect. / Streptococcus mutans é o principal agente etiológico da cárie dentária, especialmente devido à sua habilidade de adesão à superfície dentária. Esta bactéria produz glicosiltransferases, que sintetizam polímeros de glicano solúveis e insolúveis a partir da sacarose, que aumentam a colonização de bactérias cariogênicas e promovem a formação de biofilme dental na superfície dos dentes. A clorexidina é o antimicrobiano tópico mais utilizado na Odontologia, sendo considerado padrão nas diversas especialidades odontológicas, porém existem poucos estudos com esta droga em nível molecular. O objetivo do presente estudo foi investigar a atividade do gluconato de clorexidina em Streptococcus mutans UA159, plantônicos e organizados em biofilme, em diferentes concentrações e períodos de crescimento. A Concentração Inibitória Mínima (MIC) e a Concentração Bactericida Mínima (MBC) da clorexidina nas células plantônicas e MBC em biofilmes, foram determinadas pelo método da microdiluição. O RNA total das S. mutans plantônicos e organizados em biofilme, expostos ou não (controles) à clorexidina foram extraídos, purificados e reversamente transcritos a Cdna. O PCR quantitativo em tempo real foi utilizado para quantificar os níveis relativos de transcrição dos genes 16S rRNA, gtfB, gtfC e gtfD de S. mutans na presença ou ausência de clorexidina. A atividade da clorexidina na estrutura do biofilme inicial de 2 e 4h, e as alterações morfológicas nas células plantônicas, sobre concentrações variadas, foi examinada por Microscopia Eletrônica de Varredura (MEV). A MIC e a MBC para células plantônicas foram 2,2 mg/L e 18 mg/L, respectivamente, e a MBC para os biofilmes foi de 800 mg/. A exposição à clorexidina, nas concentrações de 4,5 mg/L e 9 mg/L, reduziu a contagem das células plantônicas por 6 e 20 vezes, respectivamente, enquanto a contagem das células do biofilme foram reduzidas (2,5 vezes ou mais) em concentrações acima de 500 mg/L. Nas células plantônicas, a exposição a 4.5 mg/L de CHX, aumentou a expressão das gtfC e gtfD por 11 e 4 vezes (p<0.01), respectivamente. Em biofilme, a expressão das gtfB, gtfC e gtfD foram reduzidas (<1,6 x, p<0.01) em concentrações acima de 500 mg/L. As superficies celulares aparentemente não mostraram modificação quando as células plantônicas foram expostas às concentrações de 4,5 mg/L por 2 ou 4h, mas depois de 6h, várias células murchas de S. mutans com material intracelular extravasado, foram observadas. Um decréscimo no comprimento das cadeias das células e também da matriz foram encontradas quando os biofilmes iniciais foram expostos a 1,1 mg/L e 4,5 mg/L de clorexidina, enquanto as concentrações de 1200 mg/L e 2000 mg/L causaram extensa precipitação de material desconhecido nas lamínulas. Assim, concluiu-se que os efeitos da clorexidina contra S. mutans dependem do tipo de organização celular, período de crescimento e concentração utilizada da droga. A clorexidina pode afetar a parece celular e interferir com mecanismos de formação do biofilme e, em concentrações subletais, reduz a expressão de algumas gtfs, o que pode exercer um efeito anticariogênico.

Thesis (MTech (Chemical Engineering))--Cape Peninsula University of Technology, 2012. / Recently much research and development has been done into the creation of desalination systems in South Africa, with particular emphasis on the commercialisation of desalination plants that serve local communities. This has been successful - there are currently plants running at Bitlerfontien, Bushmans River Mouth and Robben Island - although membrane fouling and scaling remains a problem associated with membrane desalination, as it does worldwide The aim of this study was to Investigate the performance of different type of antiscalants on artificially scaled membranes which we prepared inside the lab as well as on scaled membranes which were used in pilot plant. We used two type of anti-sealants in our research: Vitec 3000 and Zinc ions. The effects of these anti-sealants on the membrane were determind and the RO performances of the treated and untreated membrane compared. A suitable autopsy procedure was established and was then used to autopsy the XLE 4040 membranes. The autopsied membranes were characterized by scanning electron microscopy (SEM) and optical microscopy (OM). The SEM and OM results clearly showed that scaling had taken place: deposits were observed for both the shell and core samples, which were not present in the images of the untreated membrane, especially when looking at high SEM images magnification. Results also showed that the anti-sealants reduced the fouling and scaling on the membrane surface. As a result, the membrane rejection improved. Rejection and flux results indicated that commercial anti-sealant was more effective in preventing scaling than the Zinc ions. For Vitec anti-sealant case, the flux was in steady state at 36.8 Imh (5% less) after 5 hours compared to around 35 Imh (8% less) for zinc ions case. In addition; SEM images showed that less deposited particles are formed when the membrane was treated with commercial anti-sealant.

Orientador: Carlos Alberto de Souza Costa / Banca: Osmir Batista de Oliveira Júnior / Banca: Maria Salete Machado Cândido / Banca: Alcebíades Nunes Barbosa / Banca: Adair Luiz Stefanello Busato / Resumo: Este estudo teve como objetivo avaliar a capacidade de formação de barreira de tecido duro quando da realização de pulpotomias, onde as feridas pulpares foram capeadas com três diferentes materiais. Hidróxido de cálcio, Agregado de Trióxido Mineral (MTA), e PROROOT® MTA foram estudados quanto à morfologia e localização da barreira de tecido reparador formada, e também quanto à extensão do reparo. Três métodos de avaliação (microscopia eletrônica de varredura, lupa estereoscópica e microscópio de luz) foram empregados para analisar as imagens da barreira de tecido duro, e comparados entre si para avaliar a abrangência do espectro de leitura de cada método. Cinco cães Beagle, com idade de doze meses foram utilizados, sendo as pulpotomias realizadas nos segundos, terceiros e quartos pré-molares. Os dentes foram capeados com os materiais experimentais e controle e as cavidades restauradas com amálgama de prata. Após o período mínimo de 90 dias e máximo de 105 os animais foram sacrificados e os dentes submetidos ao processamento laboratorial para avaliação em microscopia eletrônica de varredura. Um programa analisador de imagens foi empregado para delimitar e determinar as áreas dos condutos e compará-las com as áreas das barreiras de tecido duro formadas no local. Os dados da microscopia eletrônica de varredura foram utilizados para verificar a morfologia da barreira. A localização e extensão desta estrutura reparadora foi também avaliada com auxílio de lupa esteroscópica e microscópio de luz. Os resultados obtidos permitiram demonstrar que nos espécimes pertencentes ao grupo PROROOT® ocorreu maior número de reparações completas, caracterizadas pela deposição de barreira de tecido duro, a qual apresentava característica tubular, quando comparado ao grupo hidróxido de cálcio.... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This in vivo study was carried out to evaluate the reparative potential of pulps mechanically exposed following pulpotomy with three different capping agents. Calcium hydroxide, Mineral Trioxide Aggregate (MTA), and PROROOT® MTA were evaluated regarding the morphology, location, and extension of the reparative hardbarrier deposition. Three methods were used to evaluate the images from the reparative area (scanning electron microscope - SEM, stereo microscope - Lupe, and optical microscope). The methods were compared to evaluate the reading spectrum of each applied method. Five Beagle dogs, twelve months-old were used. Pulpotomies were performed in the second, third and forth premolars. The exposed pulps were capped with the selected experimental or control materials and the cavities restored with amalgam. After 90 to 106 days the animals were sacrificed and the teeth surgically removed in bloc were processed for SEM assessment. An image analyzer was used to measure the total area of the exposition and compare it to the area occupied by the hard-barrier. Data from SEM were used to evaluate morphology and location of the hard-barrier formation as well as the percentage of pulpal wound obliteration, which was also measured by stereo microscope and optical microscope. Larger number of samples in which PROROOT® was applied as pulp-capping agent exhibited complete hard-barrier formation when compared to calcium hydroxide and MTA. For most of PROROOT® samples, the hardbarrier exhibited several dentinal tubules. On the other hand, calcium hydroxide samples presented the lowest number of total repairs and the hard-barrier observed in a few samples exhibited amorphous histological characteristics. Finally, it was demonstrated that SEM evaluation does not allow detailed assessment of the hard-barrier formation such as its extension and location... (Complete abstract, click electronic address below) / Doutor