Submitted by admin tede (tede@pucgoias.edu.br) on 2016-08-18T13:40:01Z No. of bitstreams: 1 Nathalie Borges Costa.pdf: 1419823 bytes, checksum: 9601be4b45901e83f91a699e9111d75f (MD5) / Made available in DSpace on 2016-08-18T13:40:01Z (GMT). No. of bitstreams: 1 Nathalie Borges Costa.pdf: 1419823 bytes, checksum: 9601be4b45901e83f91a699e9111d75f (MD5) Previous issue date: 2011-12-07 / Glaucoma encompasses a group of eye diseases characterized by retinal ganglion cell damage, optical nerve damage and visual field loss. It is a complex optic neuropathy of genetically heterogeneity which can lead to irreversible blindness. The visual field loss is a late manifestation symptom, therefore it is not a reliable symptom to detect glaucoma in its early stages. It is the second biggest cause of blindness worldwide, responsible for 12% of all the cases of complete vision loss, while cataracts are responsible for 48% of reversible blindness worldwide. A positive family history plays an important role as a risk factor for primary openangle glaucoma development. This is the most common form of glaucoma and it typically occurs after 40 years old of age. If the disease is neither detected nor treated properly the risk of blindness is higher. Patients with a positive family history or patients with mutations in genes that are known to be associated with glaucoma should have medical attention in order to prevent later complications. In this way it is extremely important to seek new genes that could be related to glaucoma. Up to this time three genes have been associated with primary open-angle glaucoma, the myocilin gene (MYOC), the optineurin gene (OPTN) and the repeat domain 36 gene (WDR36). The CYP1A1 gene, located at 15q22-q24, encodes an enzyme which is involved in the conversion of chemical substances into highly reactive species leading to unwanted cell damage, cell death or mutation. The m1 polimorphism is related to a high rate of enzyme activity and it comprises increased lung cancer susceptibility. The objective of this paper is to analyze the correlation of 100 patients with primary open-angle glaucoma and 52 normal controls by the RFLP method (Restriction Fragment Length polymorphism). The frequency of the homozygous wild-type (w1/w1) of CYP1A1 gene among patients with primary open-angle glaucoma (n=100) was 16%, for the genotype w1/m1 the frequency of was 77% and for m1/m1 it was 7%. Among the control group (n=52) the frequency of the homozygous wild-type (w1/w1) of CYP1A1 gene was 54%, the frequency of w1/m1 was 46% and m1/m1 was 0%. The qui-square test ( 2) considered the result statistically significant (P<0,0001), and this suggests that the CYP1A1m1 polymorphism is associated with primary open-angle glaucoma. / Glaucoma engloba inúmeras doenças oculares que têm como característica a lesão das células ganglionares da retina, com conseqüente lesão do nervo óptico e perda do campo visual. É considerada uma neuropatia óptica complexa e geneticamente heterogênea, levando à cegueira irreversível. A perda do campo visual é uma manifestação tardia do glaucoma, e portanto não é particularmente adequada para a detecção da doença em seu início. É a segunda causa de cegueira no mundo, respondendo por 12% dos casos, ficando atrás apenas da catarata (48%). A história familiar positiva é um importante fator de risco para o desenvolvimento do glaucoma primário de ângulo aberto, que é a forma mais comum da doença e que se manifesta principalmente após os 40 anos. O glaucoma, quando não diagnosticado e, portanto não tratado, pode levar a um risco maior de cegueira. Sendo assim, indivíduos com histórico familiar ou mesmo alterações nos genes associados ao glaucoma devem ter uma atenção médica mais dirigida aos seus aspectos preventivos, por isso a importância da procura de novos genes associados ao glaucoma. Até o momento foram identificados três genes relacionados ao glaucoma primário de ângulo aberto, o gene myocilin (MYOC ), o optineurin (OPTN) e o gene de domínio de repetição WD 36 (WDR36). O gene CYP1A1, localizado no cromossomo 15q22-q24, codifica enzima que está envolvida na conversão de produtos químicos em moléculas altamente reativas, que podem produzir dano celular indesejado, morte celular ou mutações. O polimorfismo m1 está associado a uma maior atividade enzimática, tendo sido referido como fator genético de suscetibilidade para o câncer de pulmão. Este trabalho teve como objetivo verificar a possível associação entre 100 doentes com glaucoma primário de ângulo aberto e 52 controles por RFLP (polimorfismo de comprimento de fragmentos de restrição). A frequência do gene CYP1A1 entre os pacientes com glaucoma primário de ângulo aberto (n=100) para o genótipo homozigoto selvagem w1/w1 foi de 16%; 77% para o genótipo w1/m1 e 7% para o genótipo m1/m1. Entre os pacientes do grupo controle (n=52) a freqüência do gene CYP1A1 foi de 54% para o genótipo w1/w1, sendo a freqüência para o genótipo w1/m1 igual a 46% e 0% para o genótipo m1/m1. Foi realizado o 2 que mostrou que o resultado deste trabalho é estatisticamente significativo (P<0,0001), sugerindo relação entre o polimorfismo CYP1A1m1 e o glaucoma primário de ângulo aberto.

Objetivos: Avaliar a capacidade do eletrorretinograma de padrão reverso (PERG) macular e multifocal (mf) de diferenciar pacientes com suspeita de glaucoma (SG) e glaucoma com defeito de campo hemianópico (GH) de controles, comparar a capacidade de discriminação do PERG e tomografia de coerência óptica (TCO) fourrier domain, e avaliar a relação entre as medidas do PERG e TCO. Métodos: Medidas do campo visual (CV) computadorizado, respostas do PERG transiente e modo estacionário e PERGmf foram obtidos dos SG (n=14, 24 olhos), GH (n=5, 7 olhos) e controles (n=19, 22 olhos). Os seguintes parâmetros de TCO foram investigados: camada de fibras nervosas da retina peripapilar (CFNRpp), espessura total da mácula e espessuras das camadas segmentadas da mácula. As medidas foram analizadas utilizando-se modelos lineares de efeito misto. Também foi avaliada a relação entre as medidas e a performance diagnóstica de cada tecnologia. Resultados: Comparado aos controles, a média do tempo de pico de P50 da resposta do PERG transiente estava reduzida nos SG e GH, enquanto que a fase, a amplitude do modo estacionário e respostas do PERGmf estavam anormais apenas no GH. A média das medidas da TCO de espessura macular e da CFNRpp nos SG e GH diferiram significativamente dos controles. Uma significativa relação foi observada entre o PERG e a maior parte dos dos parâmetros do CV central e TCO. A análise por regressão e componentes principais revelou que a TCO de nervo óptico e mácula, assim como o PERG transiente e PERGmf tiveram estatisticamente capacidade similar em discriminar os SG dos controles. Conclusões: Os parâmetros do PERG e da TCO podem estar anormais, com significativa relação entre as medidas, em uma porcentagem alta de olhos com SG com CV normal. Nossos achados sugerem que as duas tecnologias podem ser úteis e complementares na detecção precoce de glaucoma / Purpose: To evaluate the ability of macular and multifocal (mf) pattern electroretinogram (PERG) to differentiate glaucoma suspects (GS) and glaucoma with hemifield loss (GHL) from controls, to compare the discrimination ability of PERG and fourier-domain optical coherence tomography (fdOCT), and to assess the relationship between PERG and fdOCT measurements. Methods: Standard automated perimetry (SAP), steady-state and transient PERG responses and mfPERG measurements were obtained from GS (n=14, 24 eyes), GHL (n=5, 7 eyes) and controls (n=19, 22 eyes). The following fdOCT parameters were investigated: circumpapillary retinal nerve fiber layer (cpRNFL), full-thickness macula, and segmented macular layer thicknesses. Measurements were compared using mixed effects linear models. The relationships between measurements and the diagnostic performance of each technology were also assessed. Results: Compared to controls, average P50 peak time transient PERG responses were reduced in GS and GHL, whereas average phase and amplitude steady-state and mfPERG responses were abnormal only in GHL. The average fdOCT-measured cpRNFL and macular thickness measurements in GS and GHL differed significantly from controls. A significant relationship was found between PERG and most fdOCT or central SAP sensitivity parameters. Principal component regression analysis revealed that optic disc and macular OCT parameters, along with mfPERG and transient PERG parameters had statistically similar ability to discriminate GS from controls. Conclusions: PERG and OCT parameters may be abnormal, with significant correlations between measurements, in a high percentage of GS eyes with normal SAP. Our findings suggest that both technologies may be helpful and complementary in early glaucoma detection

Introdução: A cicatrização subconjuntival excessiva é a maior causa de falha cirúrgica no glaucoma. Uma vez que o ácido rosmarínico apresenta ação antifibrótica, é interessante avaliar seu uso como adjuvante na cirurgia filtrante do glaucoma. Objetivos: Comparar os achados clínicos e histopatológicos e a expressão gênica de fatores relacionados à cicatrização tecidual entre coelhos submetidos à cirurgia antiglaucomatosa experimental (CAGE) e tratados com injeções subconjuntivais de ácido rosmarínico ou com injeções de solução salina 0,9%. Material e Métodos: Estudo experimental intervencionista, do tipo ensaio clínico randomizado, no qual 41 coelhos New Zealand foram submetidos à CAGE no olho direito e divididos em dois grupos de tratamento: grupo A (n=21), tratados com injeções subconjuntivais de 0,1 ml de solução salina 0,9% e grupo B (n=20), tratados com injeções subconjuntivais de 0,1 ml de ácido rosmarínico 15 mg/ml (3 dias antes da cirurgia, ao final desta e a cada 3 dias até a eutanásia). Os efeitos do tratamento foram avaliados com cinco dias (grupos A1 e B1) e com 15 dias (grupos A2 e B2) por meio da avaliação da pressão intraocular (PIO) com Tonopen, de aspectos da ampola filtrante com base no Moorfields Bleb Grading System, do estudo histopatológico (contagem de neutrófilos/campo, vasos/campo, densidade de colágeno corado com Sirius Red, imuno-histoquímica anti--actina e anti-VEGF) e do RT-PCR (colágeno tipo 1 alfa-1 COL1A1 e TGF-2). As fotomicrografias foram avaliadas quantitativamente com o software ImageJ® 1.36 (Sirius Red, -actina e VEGF) e os dados comparados por testes não paramétricos, sendo p<0,05 considerado estatisticamente significativo. Resultados: Não houve diferença significativa nas comparações da PIO entre os grupos. Nas avaliações da ampola filtrante, apenas o grupo B2 apresentou maiores escores de área central (p=0,0004), área máxima (p=0,0007) e altura (p=0,0239). Os grupos B1 (p<0,0001) e B2 (p=0,0011) apresentaram escores significativamente menores no parâmetro vascularização que os seus respectivos controles. Houve diminuição significativa de neutrófilos/campo somente no grupo B2 (p=0,0159). O número de vasos/campo também foi significativamente menor nos grupos B1 e B2, em comparação aos seus respectivos controles (p=0,0079 e p=0,0345, respectivamente). Não foram observadas diferenças significativas na coloração com Sirius Red e na imuno-histoquímica para anti--actina entre os grupos, porém o grupo B1 apresentou menor marcação anti-VEGF (p=0,0190). As análises por RT-PCR para TGF-2 e COL1A1 não demonstraram diferenças entre os grupos, com exceção do grupo B2, que apresentou maior expressão de COL1A1 quando comparado ao seu controle (p=0,0159). Conclusões: Os resultados apontaram para os potenciais efeitos anti-inflamatórios e moduladores da neovascularização do ácido rosmarínico em cirurgias antiglaucomatosas. Apesar do grupo B demonstrar melhor aspecto clínico (exceto a PIO, que não demonstrou diferença entre os grupos), a ação antifibrótica do ácido rosmarínico não pôde ser comprovada. / Introduction: Excessive subconjunctival scarring is a major cause of surgical failure in glaucoma. Since rosmarinic acid has antifibrotic action, we evaluate its usage as an adjunctive treatment to glaucoma filtration surgery. Objectives: To compare clinical, histopathological and expression of factors related to the healing process between rabbit underwent experimental glaucoma surgery (EGS) and treated with subconjunctival injections of rosmarinic acid or with saline solution 0.9%. Methods: Interventional experimental study (randomized clinical trial) in which 41 New Zealand rabbits were submitted to EGS in the right eye and divided into two treatment groups: group A (n=21) - eyes treated with subconjunctival injections of 0.1 ml of saline solution 0.9% and B (n=20) - eyes treated with subconjunctival injections of 0.1 ml of rosmarinic acid 15mg/ml (three days prior to surgery, at the end of surgery and every three days until euthanasia). Treatment effects were assessed after five days (groups A1 and B1) and 15 days (groups A2 and B2) by evaluation of intraocular pressure (IOP) with Tonopen and bleb characteristics based on the Moorfields Bleb Grading System, as well as histopathology analysis (neutrophil/field count; vessels/field count; collagen density after stained with Sirius Red; and anti--actin and anti-VEGF immunohistochemistry) and by RT-PCR (for collagen type 1 alpha 1 - COL1A1 and TGF-2). The photomicrographs were evaluated quantitatively with ImageJ® 1.36 software (Sirius Red, -actin and VEGF) and data were compared through non-parametric tests, considering p<0.05 as statistically significant. Results: No significant difference was observed in IOP measurements between groups. Group B2 displayed significant higher scores of bleb central area (p=0.0004), maximum area (p=0.0007) and height (p=0.0239). Both B1 (p<0.0001) and B2 (p=0.0011) groups presented significantly lower vascularization scores than their respective controls. A significant decrease in neutrophils/field counting was observed only in group B2 (p=0.0159), however vessels/field counting was significantly lower in groups B1 and B2, compared to their controls (p=0.0079 and p=0.0345, respectively). No significant differences were observed in the collagen staining with Sirius Red and in the anti--actin immunohistochemistry between the all groups comparison, but group B1 displayed lower anti-VEGF measurement (p=0.0190) than group A1. RT-PCR analysis for TGF-2 and COL1A1 showed no difference between groups, except by group B2, which presented higher expression of COL1A1, compared to its control (p=0.0159). Conclusions: The findings point to the potential anti-inflammatory and anti-neovascular effects of rosmarinic acid in glaucoma surgery. Although group B show better clinical aspect (except IOP, which showed no difference between groups), the antifibrotic action of rosmarinic acid could not be proved.

INTRODUÇÃO: O blefaroespasmo essencial é uma distonia focal caracterizada por contrações involuntárias, espasmódicas e bilaterais dos músculos protratores das pálpebras. O glaucoma é a principal causa de cegueira irreversível em adultos de países desenvolvidos e a elevação da pressão intra-ocular é o maior fator de risco para o desenvolvimento da doença. A contração voluntária forçada das pálpebras pode causar aumentos da pressão intra-ocular de até 90 mmHg. Uma vez que pacientes com blefaroespasmo essencial apresentam contrações palpebrais freqüentes e de forte intensidade, eles poderiam compor um grupo de risco para o desenvolvimento do glaucoma. MÉTODOS: Vinte e oito pacientes com blefaroespasmo essencial e 28 pacientes de grupo controle, formado por indivíduos com doenças palpebrais ou no seu pós-operatório, foram submetidos a um exame oftalmológico completo. No grupo de pacientes com blefaroespasmo, a avaliação foi realizada em 8 a 11 dias após o tratamento com toxina botulínica. Efetuou-se exame de refração e medida de melhor acuidade visual corrigida com tabela de Snellen, biomicroscopia em lâmpada de fenda, tonometria de aplanação, campo visual computadorizado, teste de sobrecarga hídrica e biomicroscopia de fundo para avaliação da papila óptica. RESULTADOS: 1) A prevalência de glaucoma nos pacientes com blefaroespasmo foi significativamente maior do que nos indivíduos do grupo controle, sendo diagnosticada em 14,3% e 3,6% dos casos, respectivamente (p=0,008). 2) A pressão intra-ocular foi significativamente maior nos pacientes com blefaroespasmo (15,80 ± 3,80 mmHg) do que nos indivíduos do grupo controle (13,90 ± 2,75 mmHg) (p= 0,004). 3) O pico da pressão intra-ocular na prova de sobrecarga hídrica foi significativamente maior nos pacientes com blefaroespasmo essencial (18,82 ± 4,47 mmHg) do que nos indivíduos do grupo controle (16,27 ± 2,69 mmHg) (p=0,0421). DISCUSSÃO: Diversos estudos descreveram a influência da compressão palpebral sobre a pressão intra-ocular. A prova de sobrecarga hídrica tem sido considerada uma ferramenta indireta para se avaliar a capacidade do fluxo de drenagem do trabeculado, além de exibir correlação com os picos de pressão intra-ocular que muitas vezes não são detectados em exames de rotina. Os pacientes com blefaroespasmo essencial apresentaram pressões intra -oculares e picos de pressão intra-ocular na prova de sobrecarga hídrica mais elevados do que os pacientes do grupo controle, o que poderia indicar um baixo fluxo de drenagem. Estas altíssimas variações de pressão intra -ocular a que estes indivíduos são submetidos constantemente, em pacientes com menor fluxo de drenagem poderiam causar um aumento crônico da pressão intra -ocular e o desenvolvimento do glaucoma. CONCLUSÕES: Os resultados observados sugerem que estes pacientes compõem um grupo de risco para o desenvolvimento de glaucoma e esta doença deve ser pesquisada de maneira sistemática na avaliação inicial e durante o seguimento desses casos / INTRODUCTION: Essential blepharospasm is a focal distonia characterised by involuntary, spasmodic, bilateral contractions of eyelid protractors. Glaucoma is the most important cause of irreversible blindness in adults in developed countries and high intraocular pressure is the major risk factor for development of the disease. Voluntary forced eyelid closure can produce an intraocular pressure increase of 90 mmHg. As patients with essential blepharospasm present frequent and strong eyelid contractions, they could be at risk for glaucoma development. METHODS: Twenty eight patients with essential blepharospasm and 28 patients of a control group, with eyelid diseases or in the post-operative period, were submitted to a complete ophthalmic examination. In the group of patiens with blepharospasm, the evaluation was done between 8 to 11 days after botulinum toxin treatment. We performed refractometry and best corrected visual acuity with Snellen chart, slitlamp biomicroscopy, applanation tonometry, automated perimetry, water drinking test and dilated funduscopy to evaluate optic discs. RESULTS: 1) Prevalence of glaucoma in patients with blepharospasm (14,3%) was higher than in the individuals from control group (3,6%) (p=0,008). 2) Intraocular pressure was higher in patients with blepharospasm (15,80 ± 3,80 mmHg) than in the individuals from control group (13,90 ± 2,75 mmHg) (p=0,004). 3) Intraocular pressure peaks in the water drinking test were higher in patients with essential blepharospasm (18,82 ± 4,47 mmHg) than in the individuals from control group (16,27 ± 2,69 mmHg) (p=0,0421). DISCUSSION: Several studies reported the influence of eyelid compression over intraocular pressure. The water drinking test has been considered as an indirect tool to measure outflow facility and it has good correlation with intraocular pressure peaks which are frequently missed in routine examinations. Patients with essential blepharospasm had higher mean intraocular pressures and intraocular pressure peaks in the water drinking test than patients from control group, which could indicate low outflow facility. These high intraocular pressure variations in patients with low outflow facility could increase the intraocular pressure chronically and lead to development of glaucoma. CONCLUSION: Our results suggest that patients with essential blepharospasm be at higher risks for development of glaucoma and this disease should be always investigated at presentation and during follow-up of these cases

Peçonhas de aranhas são uma rica fonte de moléculas, dentre as quais se destacam os peptídeos neuroativos, que atuam no tecido nervoso de insetos e mamíferos, tais como receptores colinérgicos e glutamatérgicos A retina constitui um neuroepitelio, uma das membranas do segmento posterior do olho, é uma extensão do sistema nervoso central. A lesão isquêmica nesse tecido desencadeia um processo de degeneração celular, sendo os neurônios os principais afetados. Várias patologias oculares, como o glaucoma, estão associadas a uma degeneração neuronal secundária à isquemia, na qual o excesso de L-glutamato (L-Glu) extracelular é lesivo aos neurônios. A peçonha da aranha Parawixia bistriata contém componentes com grande potencial neuroprotetor, como a Parawixina10 (Pwx10), que atua potencializando o transporte de L-Glu e glicina para o meio intracelular. Neste contexto, diante da necessidade de se buscar novas terapias para o tratamento de neuropatologias e de se entender a lesão isquêmica, a Pwx10 surge como potencial fármaco neuroprotetor. Portanto, o objetivo deste trabalho foi analisar o potencial neuroprotetor da Pwx10, em um modelo de isquemia retiniana aguda, com e sem reperfusão, em ratos Wistar. Durante os experimentos de isquemia (ISQ), a pressão intra-ocular (PIO) foi aumentada para 120 mmHg, e mantida por 45 minutos. Nos experimentos em que houve reperfusão (ISQ/REP), após a isquemia, a pressão foi reduzida aos níveis normais e mantida por mais 15 minutos, de forma a restaurar o fluxo sanguíneo e os níveis basais da PIO. As drogas utilizadas para tratamento foram injetadas por via intravitrea, 15 minutos antes do início da isquemia. Após a cirurgia foram realizados os processos histológicos que envolvem técnicas de H-E e Fluoro-Jade C. Em seguida, foram analisadas as densidades de células viáveis na camada nuclear interna (CNI) e camada de células ganglionares (CCG). Os resultados mostraram que os tratamentos com a Pwx10 protegeram as células da CNI tanto em ISQ como ISQ/REP. Comparada com o Riluzole, a Pwx10 foi mais eficaz em CCG ISQ em 15% e CNI ISQ/REP em 23%. Portanto, a Pwx10 apresenta efeitos neuroprotetores em ratos Wistar submetidos à isquemia retiniana aguda, seguida por reperfusão ou não. / Spider venoms are a rich source of molecules, among which stand out the neuroactive peptides that act on the nervous tissue of insects and mammals, such as cholinergic and glutamatergic receptors. The retina is a neuroepithelium, a membrane lining the cavity of the eyeball, it being an extension of the central nervous system. Ischemic injury that tissue triggers a cell degeneration process, and the neurons affected major. Various eye diseases such as glaucoma, are associated with neuronal degeneration secondary to ischemia in which excess L-glutamate (L-Glu) extracellular is harmful to neurons. The venom of Parawixia bistriata spider contains components with high neuroprotective potential, as Parawixina10 molecule (Pwx10), which operates enhancing the transport of L-Glu and glycine to the intracellular medium. In this context, on the need to seek new therapies for the treatment of these diseases and to understand the ischemic injury, Pwx10 emerges as potential neuroprotective drug. Therefore the aim of this study was to evaluate the neuroprotective potential of Pwx10 on an acute retinal ischemia model, with and without reperfusion in rats. During the experiments ischemia (ISC), the intraocular pressure (IOP) was increased to 120 mmHg and maintained at this level for 45 minutes. In experiments in which there was reperfusion (I/R) after the period of ischemia, the pressure was reduced to normal levels and maintained there for 15 minutes in order to restore blood flow and baseline IOP. The drugs used for the treatment were intravitreally injected 15 minutes before the onset of ischemia. After surgery were performed histological techniques involving procedures H-E and Fluoro-Jade C. Then, viable cell densities in the inner nuclear layer were analyzed (INL) and ganglion cell layer (GCL). The results showed that the treatments with Pw10 protected the INL cells both in ISC as IR. Compared with Riluzole, the Pwx10 was more effective in GCL ISC 15% and INL I/R 23%. Therefore, Pwx10 shows neuroprotective effects in Wistar rats with acute retinal ischemia followed by reperfusion or not.

青光眼作為一種慢性進展性視神經病變，已經成為世界眼科病變中導致不可逆盲的首要原因。青光眼的早期診斷和治療對於降低疾病進展的風險至關重要。光學相干斷層掃描（OCT）可以提供在體視網膜橫斷面的視圖，從而實現了對視網膜神經纖維層（RNFL）改變的客觀測量，這些改變已經被證明了與青光眼引起的視神經損害相關，並已成為診斷青光眼的重要參考依據。 / 頻域OCT是最新一代的光學相干斷層掃描，它具有比時域OCT更快的掃描速度和更高的圖像解析度，因此，頻域OCT可以提供更可靠的RNFL厚度測量和RNFL缺損評估。本文的研究目的在於評估頻域OCT對RNFL厚度的重測再現性，以及探討影響RNFL厚度測量的因素，這些因素包括（1）影像平均法的應用，（2）RNFL分層錯誤，和（3）視網膜血管的影響。此外，由於RNFL攝影是一個評估青光眼RNFL缺損的臨床參考標準，我們還將其對RNFL缺損的測量與頻域OCT的RNFL厚度偏差圖所作出的測量進行了比較。 / 首先，為了評估頻域OCT對RNFL厚度測量的重測再現性，15名正常人和15名青光眼患者連續四周每週均接受一次OCT掃描。正常組和青光眼組的RNFL厚度再現性係數分別為4.77-12.65微米和4.53-16.66微米，由於組內相關性係數均大於0.773，說明頻域OCT所作出的RNFL厚度測量是具備可重複性的。 / 其次，通過分析54隻眼（25名正常志願者和29名青光眼患者）的RNFL厚度測量值，本文對圖像平均法的應用是否會影響RNFL厚度的測量這一問題進行了探討。分析中，每一隻眼均接受了3次OCT掃描，3次掃描的圖像分別使用2、8、和16張連續的圖像進行影像平均。結果顯示，除了青光眼組的鼻下象限RNFL厚度測量值之外（P=0.036），不同的圖像幀數並不會對兩組的總體和其它各象限的RNFL厚度測量值產生顯著的影響（P≥0.055）。雖然圖像平均法的應用對RNFL厚度測量的影響並不顯著，但是視網膜血管和RNFL分層錯誤對青光眼,尤其是對RNFL非常薄的晚期青光眼患者的RNFL厚度測量有影響。結論來自對60個正常人，66個輕至中度青光眼（MD≥-6 dB）患者和54個嚴重青光眼（MD<-6 dB）患者的共180張OCT圖像的分析。視網膜血管相對於平均RNFL厚度的比例均值在正常組，輕至中度青光眼組，和嚴重青光眼組分別為11.2±2.3，12.6±2.5，和16.6±3.9。在人為調整了RNFL界限以糾正RNFL分層錯誤的前後，總體RNFL厚度的差異範圍在正常組為-3.0-2.5微米，輕至中度青光眼組為-2.5-5.0微米，嚴重青光眼組為-11.0-9.5微米組。 / 最後，通過對41名青光眼患者的51隻眼的RNFL缺損面積，位置，和覆蓋角度進行測量，本文將頻域OCT作出的測量結果和共焦鐳射掃描檢眼鏡（CSLO）RNFL反射影像圖的測量結果進行了比較，結果顯示：OCT不但可以檢測到所有出現在CSLO的RNFL反射影像圖上的RNFL缺損，更重要的是，OCT還可以檢測出額外的並未在RNFL反射影像圖上出現的RNFL缺損。 / 總之，頻域OCT是一種可提供高再現性RNFL厚度測量的影像方法。對青光眼,尤其是晚期青光眼的RNFL厚度測量值的詮釋，應當考慮到視網膜血管和RNFL分層錯誤的影響。OCT具備對RNFL缺損進行多維度量化（包括厚度，面積，位置，和覆蓋角度）的能力，在青光眼RNFL改變的檢測和監測方面，相對于傳統的RNFL攝影，OCT無疑是更有效的選擇。 / Glaucoma, a chronic progressive optic neuropathy, is the leading cause of irreversible blindness in the world. An early diagnosis and treatment of glaucoma is vital to reduce the risk of disease progression. Providing a cross-sectional view of the retina in vivo, optical coherence tomography (OCT) can objectively measure the changes of retinal nerve fiber layer (RNFL), which has been shown to be of relevance and importance in detecting glaucomatous damage of the optic nerve. / The latest generation of OCT, the spectral-domain OCT, has a faster scan speed and a higher image resolution compared to the time-domain OCT. It is expected that the spectral-domain OCT would allow a more reliable measurement of the RNFL thickness and assessment of RNFL defects. The objectives of this research project were to examine the test-retest reproducibility of spectral-domain OCT RNFL measurement and investigate factors including (1) image averaging, (2) segmentation failure, and (3) contribution of retinal blood vessels that might affect the measurement of RNFL thickness. As RNFL photography is a reference standard to evaluate RNFL defects in glaucoma, we also evaluated whether RNFL defects measured in the spectral-domain OCT RNFL thickness map would be comparable to those detected in RNFL photographs. / To evaluate the test-retest reproducibility of RNFL measurements obtained by the spectral-domain OCT, 15 normal individuals and 15 glaucoma patients were followed and imaged weekly for 4 consecutively weeks. The reproducibility coefficients of RNFL thicknesses ranged between 4.53 and 16.66 μm for the normal group, and 4.77 and 12.65 μm for the glaucoma group. The intraclass correlation coefficients were all above 0.773, indicating RNFL measurement with spectral-domain OCT was reproducible. / We then investigated if multiple-image averaging would influence the measurement of RNFL thickness. A total of 54 eyes from 25 normal volunteers and 29 glaucoma patients with RNFL images captured and averaged with 2, 8, and 16 consecutive image frames were analyzed. For both groups, there were no significant differences in global or sectoral RNFL thicknesses among the image series averaged with different number of image frames (all with P≥0.055) except for the inferonasal sector in the glaucoma group (P=0.036). Although the impact of image averaging on RNFL measurement was insignificant, the presence of retinal blood vessels and segmentation errors were influential on the measurement, particularly in advanced glaucoma patients when the RNFL was thin. Analyzing a total of 180 eyes from 60 normal individuals, 66 mild to moderate (MD≥-6 dB) and 54 advanced (MD<-6 dB) glaucoma patients, the mean proportion of retinal blood vessels relative to the average RNFL thickness was 11.2±2.3%, 12.6±2.5% and 16.6±3.9%, respectively. After correcting the segmentation errors by manually refining the RNFL boundaries, the differences in average RNFL thickness ranged from -3.0 to 2.5 m in the normal, -2.5 to 5.0 m in the mild to moderate glaucoma and -11.0 to 9.5 m in the advanced glaucoma groups. / Finally, we compared the area, the angular location, and the angular width of RNFL defects from 51 eyes of 41 glaucoma patients measured with the spectral-domain OCT and RNFL reflectance images obtained by a confocal scanning laser ophthalmoscope (CSLO). OCT was able to detect areas of RNFL abnormalities in all eyes with RNFL defects which were evident in the CSLO RNFL reflectance images. More important, OCT could identify additional RNFL thinning not apparent in RNFL reflectance images. / In summary, spectral-domain OCT could offer an effective approach in measuring RNFL with high reproducibility. Interpretation of RNFL measurement should take the contribution of the retinal blood vessels and segmentation errors into consideration, particularly in advanced glaucoma when the RNFL is thin. With the ability to quantify multiple dimensions of RNFL defects (thickness, area, angular location, and angular width), OCT could provide a useful alternative to detect and monitor RNFL changes in glaucoma. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Ye, Cong. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 117-130). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / ABSTRACT --- p.i / 摘要 (ABSTRACT IN CHINESE) --- p.v / DEDICATION --- p.viii / ACKNOWLEDGEMENT --- p.ix / TABLE OF CONTENTS --- p.x / PUBLICATIONS --- p.xiv / ABBREVIATIONS --- p.xvi / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter 1.1 --- Glaucoma --- p.2 / Definition of Glaucoma --- p.2 / Epidemiology of Glaucoma --- p.3 / Pathogenesis of Glaucoma --- p.4 / Diagnosis of Glaucoma --- p.7 / Chapter 1.2 --- Retinal Nerve Fiber Layer --- p.13 / Anatomy of Retinal Nerve Fiber Layer --- p.13 / Visualization of Retinal Nerve Fiber Layer --- p.14 / Retinal Nerve Fiber Layer Defect in Glaucoma --- p.16 / Significance of Detecting Retinal Nerve Fiber Layer Defect in Glaucoma --- p.18 / Chapter 1.3 --- Optical Coherence Tomography --- p.20 / Principle of Optical Coherence Tomography --- p.20 / Retinal Nerve Fiber Layer Imaging with OCT --- p.21 / Optic Nerve Head Imaging with OCT --- p.27 / Advantages and Disadvantages of Optical Coherence Tomography --- p.29 / Chapter 1.4 --- Research Objectives --- p.30 / Chapter CHAPTER 2: --- GENERAL MATERIALS AND METHODS --- p.32 / Chapter 2.1 --- Subject Enrollments --- p.33 / Chapter 2.2 --- Clinical Ophthalmic Examination --- p.34 / Chapter 2.3 --- Visual Field Examination --- p.35 / Definition of Normal and Glaucoma Groups --- p.35 / Chapter 2.4 --- Optical Coherence Tomography Imaging --- p.37 / Cirrus HD-OCT Imaging --- p.37 / Spectralis OCT Imaging --- p.37 / Chapter 2.5 --- Statistical Analysis --- p.39 / Chapter CHAPTER 3: --- RETINAL NERVE FIBER LAYER IMAGING WITH SPECTRAL-DOMAIN OPTICAL COHERENCE TOMOGRAPHY --- p.40 / Chapter 3.1 --- Reproducibility and Agreement of Retinal Nerve Fiber Layer Measurement --- p.41 / Introduction and Study Objectives --- p.41 / Methods --- p.42 / Results --- p.45 / Discussion --- p.47 / Tables and Figures --- p.51 / Chapter 3.2 --- Effect of Multiple B-scans Averaging on Retinal Nerve Fiber Layer Measurement --- p.58 / Introduction and Study Objectives --- p.58 / Methods --- p.59 / Results --- p.61 / Discussion --- p.62 / Tables and Figures --- p.67 / Chapter 3.3 --- Impact of Blood Vessels and Segmentation Failure on Retinal Nerve Fiber Layer Measurement --- p.73 / Introduction and Study Objectives --- p.73 / Methods --- p.75 / Results --- p.78 / Discussion --- p.80 / Tables and Figures --- p.84 / Chapter 3.4 --- Agreement of Localized Retinal Nerve Fiber Layer Defect Assessment with Confocal Scanning Laser Ophthalmoscopy --- p.95 / Introduction and Study Objectives --- p.95 / Methods --- p.97 / Results --- p.101 / Discussion --- p.103 / Tables and Figures --- p.108 / Chapter CHAPTER 4: --- GENERAL CONCLUSIONS --- p.115 / REFERENCES --- p.117

Leung Yuk Fai. / "June 2002." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (p. 132-166). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Submitted by admin tede (tede@pucgoias.edu.br) on 2017-09-04T12:48:49Z No. of bitstreams: 1 EDUARDO EUSTÁQUIO DE ALMEIDA FILHO.pdf: 1586241 bytes, checksum: ac6c2a57234dbdb8dcad9ea043e60dee (MD5) / Made available in DSpace on 2017-09-04T12:48:49Z (GMT). No. of bitstreams: 1 EDUARDO EUSTÁQUIO DE ALMEIDA FILHO.pdf: 1586241 bytes, checksum: ac6c2a57234dbdb8dcad9ea043e60dee (MD5) Previous issue date: 2017-06-12 / Glaucoma can be defined as a progressive degeneration of the retinal nerve fiber layer that results in death of the ganglion cells, eventually causing blindness. It is the first cause of irreversible blindness in the world. The pathophysiology of glaucoma is not fully elucidated, although several risk factors such as genetic, racial, age and increased intraocular pressure are known. This study was a case-control study in which 116 samples were analyzed: 32 patients with primary open-angle glaucoma (18 males and 14 females; age range 65-89 years) and 84 controls (44 males and 40 females; Aged 65-86 years). The samples were submitted to DNA extraction, then to PCR and analyzed on 2% agarose gel, stained with 5 g / mL ethidium bromide. The G894T polymorphism of the eNOS gene was found in the 0% (0/32) group of homozygotes for the wild-type (GG) allele, 93.75 % (30/32) of heterozygotes (GT) and 6.25% (2/32) homozygous for the polymorphic (TT) allele. In the control group, 9.52% (8/84) of homozygotes for the wild-type (GG), 82.14% (69/84) heterozygotes (GT) and 8.33% (7/84) homozygotes for the polymorphic allele (TT). There was no statistically significant difference. As for the allelic frequency of the eNOS gene (G894T) in the case group, 46.8% of the (G) allele and 53,2% of the T allele were found, whereas the G allele frequency was 50.5% and 49, 5% of the T allele. There was no statistically significant difference. Likewise, there was no statistically significant relationship between the polymorphism with hypertension, gender, diabetes and ethnicity. There are many possible interactions of these polymorphisms and the development of glaucoma, but further studies are needed to further elucidate these associations. / Glaucoma pode ser definido como uma degeneração progressiva da camada de fibras nervosas da retina que resulta na morte das células ganglionares, eventualmente causando cegueira. É a primeira causa de cegueira irreversível no mundo. A fisiopatologia do glaucoma não é totalmente elucidada, embora se tenha conhecimento de vários fatores de risco como alterações genéticas, raciais, idades e aumento da pressão intraocular. Este estudo trata-se de um casocontrole onde foram analisadas 116 amostras sendo 32 pacientes com glaucoma primário de ângulo aberto (18 homens e 14 mulheres; variação de idade 65-89 anos) e 84 controles (44 masculinos e 40 femininos; variação de idade 65-86 anos). As amostras foram submetidas à extração de DNA, em seguida à PCR e analisadas em gel de agarose a 2%, corados com brometo de etídio a 5 g/mL. Na investigação do polimorfismo G894T do gene eNOS foi encontrado no grupo caso 0% (0/32) de homozigotos para o alelo selvagem (GG), 93,75% (30/32) de heterozigotos (GT) e 6,25% (2/32) homozigotos para o alelo polimórfico (TT). No grupo controle foi encontrado 9,52% (8/84) de homozigotos para o alelo selvagem (GG), 82,14% (69/84) de heterozigotos (GT) e 8,33% (7/84) de homozigotos para o alelo polimórfico (TT). Não houve diferença estatística significante. Quanto a frequência alélica do gene eNOS (G894T) no grupo caso foi encontrado 46,8% do alelo selvagem (G) e 53,2% do alelo T, já no grupo controle a frequência do alelo G foi de 50,5% e 49,5% do alelo T. Não houve diferença estatística significante. Da mesma forma não houve relação estatisticamente significativa do polimorfismo com hipertensão arterial, gênero, diabetes e etinia. São muitas as possíveis interações desses polimorfismos e o desenvolvimento de glaucoma, mas ainda são necessários mais estudos para uma maior elucidação destas associações.

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 115-121).

Myocilin, expressed in the trabecular meshwork of the eye, has been linked to inherited primary open-angle glaucoma (POAG). The biological function of myocilin is unknown, but mutant myocilin exhibits a gain-of-function mechanism, aggregating within the endoplasmic reticulum of human trabecular meshwork cells, causing cell stress and eventually apoptosis. After apoptosis occurs, the trabecular meshwork is compromised, leading to an increase in intraocular pressure, a symptom of glaucoma. In this thesis, I have expressed and purified the wild-type olfactomedin (OLF) domain and 24 reported disease-causing variants. I developed a facile thermal stability assay using differential scanning fluorimetry, which follows the unfolding of a protein through the fluorescence of a dye sensitive to hydrophobic regions of a protein. Also in this thesis I have determined melting temperatures for the wild-type and for each of the disease-causing mutants. I have tested the stability of the mutants in the presence of seven osmolytes, with sarcosine and trimethylamine-N-oxide restoring the melting temperature closest to wild-type. Additionally, I expressed and purified three reported single nucleotide polymorphisms (SNPs) (E352Q, E396D, K398R), which are considered benign variants. Variants were also compared by circular dichroism, revealing high b-sheet content and wild-type structure. When compared to previous studies, there is a positive correlation between the melting temperature, and previously reported qualitative assays, which measure the mutant myocilin solubility in detergent, secretion from mammalian cells, and aggregation propensity. Taken together, these data give insight into the relationship between glaucoma genotypes and phenotypes.