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Thesis (Ph. D.)--University of Missouri--Columbia, 2003. / "May 2003." Typescript. Vita. Includes bibliographical references (leaves 139-154).

Thesis (Ph. D.) -- University of Texas Southwestern Medical Center at Dallas, 2003. / Vita. Bibliography: 160-174.

PURPOSE. CD36 is a Class B scavenger receptor that is constitu- tively expressed in the corneal epithelium and has been impli- cated in many homeostatic functions, including the homeosta- sis of the epidermal barrier. The aim of this study is to determine (1) whether CD36 is required for the maintenance of the corneal epithelial barrier to infection, and (2) whether CD36-deficient mice present with an increased susceptibility to bacterial keratitis. METHODS. The corneas of CD36- /- , TSP1- /- , TLR2- /- , and C57BL/6 WT mice were screened via slit lamp microscopy or ex vivo analysis. The epithelial tight junctions and mucin layer were assessed via LC-biotin and Rose Bengal staining, respec- tively. Bacterial quantification was performed on corneal but- tons and GFP-expressing Staphylococcus aureus was used to study bacterial binding. RESULTS. CD36-/- mice develop spontaneous corneal defects that increased in frequency and severity with age. The mild corneal defects were characterized by a disruption in epithelial tight junctions and the mucin layer, an infiltrate of macro- phages, and increased bacterial binding. Bacterial quantifica- tion revealed high levels of Staphylococcus xylosus in the corneas of CD36-/- mice with severe defects, but not in wild-type controls. CONCLUSIONS. CD36 -/- mice develop spontaneous bacterial keratitis independent of TLR2 and TSP1. The authors conclude that CD36 is a critical component of the corneal epithelial barrier, and in the absence of CD36 the barrier breaks down, allowing bacteria to bind to the corneal epithelium and result- ing in spontaneous keratitis. This is the first report of sponta- neous bacterial keratitis in mice. (Invest Ophthalmol Vis Sci. 2011;52:256–263) DOI:10.1167/iovs.10-5566

Thesis (Ph. D.)--University of Missouri--Columbia, 2003. / Typescript. Vita. Includes bibliographical references (leaves 139-154).

Orientador: Alexandre Lima Andrade / Banca: Flávia de Almeida Lucas / Banca: Silvia Maria Caldeira Franco Andrade / Resumo: As ceratites ulcerativas são comuns na rotina do médico veterinário de pequenos animais, visto que a córnea é uma estrutura vulnerável devido sua exposição ao meio externo. Por compreender uma das estruturas responsáveis pela refração luminosa e participante do mecanismo de formação da imagem, é de extrema importância a integridade da anatomia e transparência da córnea para esta continuar desempenhando suas funções. Estudos ainda são realizados com o intuito de viabilizar opções terapêuticas cirúrgicas para o tratamento de úlceras de córnea complicadas, tentando alcançar uma cicatrização com um menor dano à estrutura corneal e menor opacidade cicatricial, devolvendo a transparência da córnea. Visto que os polímeros biodegradáveis tem se mostrado alvo de pesquisas atuais para aplicações médicas, com este estudo objetivou-se avaliar originalmente a biocompatibilidade da membrana do polímero poli(butileno adipato-co-tereftalato) (PBAT) na córnea e posterior aplicação clínica deste material nas ceratites ulcerativas profundas. Para tal, foi realizada avaliação clínica experimental e histopatológica após enxertia interlamelar do PBAT em córnea de coelhos, ao longo de 60 dias, e acompanhamento da evolução clínica do emprego da membrana de PBAT no tratamento de ceratites ulcerativas complicadas em cães, comparando ao uso do enxerto conjuntival, ao longo de 90 dias. Devido aos baixos sinais de inflamação corneal clínicos e histopatológicos nos coelhos e a cicatrização corneal em todo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Ulcerative keratitis are common in the routine of a small animals veterinary doctor, given that the cornea is a vulnerable structure due to its exposure to external environment. For containing one of the structures responsible for light refraction and participant in the mechanism of formation of images, it is of extreme importance the integrity and transparency of the cornea in order for it to continue to develop its functions. Studies are still performed with the intent of making feasible surgical therapeutic options for the treatment of complicated corneal ulcers, trying to reach cicatrization with less damage to the corneal structure and less scar opacity, seeking corneal transparency. Given that biodegradable polymers have been target of current research for medical applications, the objective of this study was to evaluate originally the biocompatibility of the poly (butylene adipate-co-terephthalate) polymer (PBAT) in the cornea and posterior clinical application of this material in deep ulcerative keratitis. It was performed clinical and histopathological evaluation after interlamellar grafting of PBAT in the cornea of rabbits, over 60 days, and follow-up of the clinical evolution of PBAT membrane in the treatment of complicated ulcerative keratitis in dogs, comparing to the use of conjunctival graft, over 90 days. Due to low levels of clinical and histopathological corneal inflammation found in rabbits and corneal cicatrization in all dogs, without occurence of visual ... (Complete abstract click electronic access below) / Mestre

O tratamento da ceratite por Acanthamoeba é longo, inespecífico e de baixa adesão do paciente. Assim, a busca por novas estratégias de tratamento é necessária. Extratos não aquosos de algumas espécies de Pterocaulon exibem efeito antimicótico e antiparasitário, sendo esta atividade correntemente relacionada à presença de cumarinas. Neste contexto, o presente estudo teve por objetivo o desenvolvimento de nanoemulsões contendo extrato hexânico de Pterocaulon balansae Chodat (rico em cumarinas) visando à obtenção de um produto de uso ocular com atividade amebicida. Em uma primeira etapa, o extrato hexânico foi caracterizado por cromatografia líquida de alta eficiência com detector de arranjo de diodos, detectando-se a presença de quatro cumarinas majoritárias, entre elas, a 5-metoxi-6,7-metileno dioxicumarina (5MMDC), selecionada como marcador químico do extrato vegetal. O extrato foi na sequência incorporado em nanoemulsões constituídas de um núcleo de triglicerídeos de cadeia média estabilizado por lecitina de gema de ovo, preparadas por emulsificação espontânea. Tal procedimento conduziu à obtenção de nanoemulsões monodispersas com diâmetro médio de gotícula de aproximadamente 200-300 nm, independente da quantidade de extrato adicionado à formulação (1,0 a 5,0 mg/mL). A atividade amebicida das formulções contra Acanthamoeba castellanii foi dependente da dose e do tempo de incubação, sendo 24 horas e a concentração de 1,25 mg/mL de extrato considerada como ótima (~5% de viabilidade), com efeito similar ao controle clorexidina. Enfim, os estudos de citotoxicidade in vitro demonstraram que as células de epitélio de córnea humana (HCE) não foram afetadas com a incubação com as nanoemulsões através do ensaio de MTT. Esses resultados sugerem o potencial do extrato hexânico, rico em cumarinas de Pterocaulon balansae, associado a nanoemulsões como uma nova estratégia para o tratamento da ceratite ocular causada por Acanthamoeba. / Treatment for keratitis caused by Acanthamoeba is unspecific, long term and with low patient compliance, being the search of new treatment strategies a need. Non-aqueous extracts of some species of Pterocaulon exhibit antimycotic and anti-parasitic activity, usually attributed to the presence of coumarins. In this scenario, the aim of this work is the development of nanoemulsion to associate the coumarin-rich n-hexane extract of Pterocaulon balansae. Previously, the extract was prepared and characterized by high performance liquid chromatography with photodiode array detector. The presence of 4 major coumarins was detected, where 5-methoxy-6,7-methylene dioxycoumarin (5MMDC) was selected as chemical marker. This extract was associated to nanoemulsions composed of egg lecithin and medium chain triglycerides, prepared by spontaneous emulsification. The physicochemical characterization showed the formation of monodisperse nanoemulsions with 200-300 nm diameter, regardless the amount of extract incorporated (1.0-5.0 mg/mL). The amoebicidal activity of the formulations against Acanthamoeba castellanii was both dose-dependent and incubation time-dependent, being 24h of incubation and concentration of 1.25 mg/mL of the extract the optimal (~5% viability), with effect similar to chlorexidine control. Finally, in vitro cytotoxicity studies showed that human corneal epithelial cells were unaffected after incubation with nanoemulsions by MTT assay. These results suggest a potential of the coumarin-rich n-hexane extracts of Pterocaulon balansae associated to nanoemulsion as a new strategy for the treatment of ocular keratitis caused by Acanthamoeba.

Acanthamoeba are ubiquitous protists that play an environmental role in regulating microbial diversity; they also occasionally cause infections of the eye (Acanthamoeba keratitis) and brain (granulomatous amoebic encephalitis). These organisms exhibit two distinct phenotypes. The trophozoite form dominates in favorable conditions, in which the Acanthamoeba move through the extension of pseudopodia, engulfing microbes and other particles. During stressful conditions, the Acanthamoeba undergo a process of encystment, in which they build a double cell wall and become relatively inactive. The cyst form can survive years until more favorable conditions arise, at which point they may excyst. For this study, multiple laboratory encystment methods were compared to determine the percent encystment and the different viabilities of laboratory-produced cysts. Furthermore, four different encystment genes were targeted for development of a primer library for reverse-transcription, polymerase chain reaction expression studies. The library was developed using sequences accessed from various databases, including NCBI and EMBL; primers were screened through polymerase chain reaction, and those primers producing positive results were used to further screen cellular RNA that was extracted from encysting cells over various time points during the encystment process, and using various encystment media. Using these methods, target gene involvement in the encystment process was compared between species and encystment methods. These studies lay the foundation for quantitative gene expression analysis, and provide the basis for comparison of various encystment methods.

Thesis (Ph. D.)--School of Pharmacy and Dept. of Chemistry. University of Missouri--Kansas City, 2007. / "A dissertation in pharmaceutical sciences and chemistry." Advisor: Ashim K. Mitra. Typescript. Vita. Description based on contents viewed July 16, 2008; title from "catalog record" of the print edition. Includes bibliographical references (leaves 173-182). Online version of the print edition.

Submitted by Aline Cardoso Pereira (aline.cardosopereira@hotmail.com) on 2018-08-02T22:27:37Z No. of bitstreams: 1 DISSERTAÇÃO DE MESTRADO - ALINE C. PEREIRA.pdf: 2161021 bytes, checksum: 07880c29666b475277e0461208a4bf5a (MD5) / Approved for entry into archive by Isabel Pereira de Matos (isabel@fmva.unesp.br) on 2018-08-03T18:51:33Z (GMT) No. of bitstreams: 1 pereira_ac_me_araca_int.pdf: 2161021 bytes, checksum: 07880c29666b475277e0461208a4bf5a (MD5) / Made available in DSpace on 2018-08-03T18:51:33Z (GMT). No. of bitstreams: 1 pereira_ac_me_araca_int.pdf: 2161021 bytes, checksum: 07880c29666b475277e0461208a4bf5a (MD5) Previous issue date: 2018-06-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As ceratites ulcerativas são comuns na rotina do médico veterinário de pequenos animais, visto que a córnea é uma estrutura vulnerável devido sua exposição ao meio externo. Por compreender uma das estruturas responsáveis pela refração luminosa e participante do mecanismo de formação da imagem, é de extrema importância a integridade da anatomia e transparência da córnea para esta continuar desempenhando suas funções. Estudos ainda são realizados com o intuito de viabilizar opções terapêuticas cirúrgicas para o tratamento de úlceras de córnea complicadas, tentando alcançar uma cicatrização com um menor dano à estrutura corneal e menor opacidade cicatricial, devolvendo a transparência da córnea. Visto que os polímeros biodegradáveis tem se mostrado alvo de pesquisas atuais para aplicações médicas, com este estudo objetivou-se avaliar originalmente a biocompatibilidade da membrana do polímero poli(butileno adipato-co-tereftalato) (PBAT) na córnea e posterior aplicação clínica deste material nas ceratites ulcerativas profundas. Para tal, foi realizada avaliação clínica experimental e histopatológica após enxertia interlamelar do PBAT em córnea de coelhos, ao longo de 60 dias, e acompanhamento da evolução clínica do emprego da membrana de PBAT no tratamento de ceratites ulcerativas complicadas em cães, comparando ao uso do enxerto conjuntival, ao longo de 90 dias. Devido aos baixos sinais de inflamação corneal clínicos e histopatológicos nos coelhos e a cicatrização corneal em todos os cães, sem ocorrência de déficit visual, com o presente estudo acreditamos ter viabilizado mais uma opção terapêutica para as ceratites ulcerativas na medicina veterinária. / Ulcerative keratitis are common in the routine of a small animals veterinary doctor, given that the cornea is a vulnerable structure due to its exposure to external environment. For containing one of the structures responsible for light refraction and participant in the mechanism of formation of images, it is of extreme importance the integrity and transparency of the cornea in order for it to continue to develop its functions. Studies are still performed with the intent of making feasible surgical therapeutic options for the treatment of complicated corneal ulcers, trying to reach cicatrization with less damage to the corneal structure and less scar opacity, seeking corneal transparency. Given that biodegradable polymers have been target of current research for medical applications, the objective of this study was to evaluate originally the biocompatibility of the poly (butylene adipate-co-terephthalate) polymer (PBAT) in the cornea and posterior clinical application of this material in deep ulcerative keratitis. It was performed clinical and histopathological evaluation after interlamellar grafting of PBAT in the cornea of rabbits, over 60 days, and follow-up of the clinical evolution of PBAT membrane in the treatment of complicated ulcerative keratitis in dogs, comparing to the use of conjunctival graft, over 90 days. Due to low levels of clinical and histopathological corneal inflammation found in rabbits and corneal cicatrization in all dogs, without occurence of visual deficit, with this study we believe it was made feasible another therapeutic option for ulcerative keratitis in veterinary medicine.