Glaucoma is the second leading cause of irreversible blindness in the United States and is the leading cause of blindness in African Americans. Cupping or excavation of the optic nerve, which sends the visual signal from the photoreceptors in the eye to the brain, is a chief feature of glaucoma. A similar excavated appearance of the optic nerve is also the primary clinical sign of other congenital malformations of the eye including optic nerve head coloboma, optic pit, and morning glory disc anomaly collectively termed cavitary optic disc anomaly (CODA). Clinical similarities between CODA and glaucoma have suggested that these conditions may have overlapping pathophysiology. Although risk factors are known, such as the elevated intraocular pressure (IOP) observed in some glaucoma subjects, the biological pathways and molecular events that lead to excavation of the optic disc in glaucoma and in CODA are incompletely understood, which has hindered efforts to improve diagnosis and treatment of these diseases. Consequently, there is a critical need to clarify the biological mechanisms that lead to excavation of the optic nerve, which will lead to improvements in our understanding of these important disease processes. Because of their similar clinical phenotypes and the limited therapy geared at lowering IOP in glaucoma patients, our central hypothesis is that genes involved in Mendelian forms of CODA would also be involved in a subset of glaucoma cases and may provide insight into glaucomatous optic neuropathy. The purpose of my research project has been to identify and functionally characterize the gene that causes congenital autosomal dominant CODA in a multiplex family with 17 affected members. The gene that causes CODA was previously mapped to chromosome 12q14 and following screening of candidate genes within the region that did not yield any plausible coding sequence mutations, a triplication of a 6KB segment of DNA upstream of the matrix metalloproteinase 19 (MMP19) gene was subsequently identified using comparative genomic hybridization arrays and qPCR. This copy number variation (CNV) was present in all affected family members but absent in unaffected family members, a panel of 78 normal control subjects, and the Database of Genomic Variants. In a case-control study of singleton CODA subjects, CNVs were also detected; we detected the same 6KB triplication in 1 of 24 subjects screened. This subject was part of another 3-generation autosomal dominant CODA pedigree where affected members each have the same CNV identified in the larger CODA pedigree. A separate case-control study with 172 glaucoma cases (primary open angle glaucoma = 84, normal tension glaucoma = 88) was evaluated for MMP19 CNVs, however none were detected. Although our cohort of CODA patients is small limiting our ability to accurately determine the proportion of CODA caused by MMP19 mutations, our data indicates that the MMP19 CNV is not an isolated case and additional CODA subjects may have MMP19 defects. Because of the location of the CNV, we evaluated its effect on downstream gene expression with luciferase reporter gene assays. These assays revealed that the 6KB sequence spanned by the CNV in CODA subjects functioned as a transcriptional enhancer; in particular, a 773bp segment had a strong positive influence (8-fold higher) on downstream gene expression. MMP19, a largely understudied gene, was further characterized by expression studies in the optic nerve and retina. Using frozen sections from normal donor eyes, we demonstrated that MMP19 is predominantly localized to the optic nerve head in the lamina cribrosa region with moderate labeling in the postlaminar region, and weak labeling in the prelaminar region and retina. We also evaluated MMP19 expression in relation to the cell types that populate the optic nerve such as astrocytes and retinal ganglion cells. The pattern of expression is consistent with MMP19 being a secreted protein accumulating in the extracellular spaces and basement membranes of the optic nerve. Our studies have identified the first gene associated with CODA and future research is focused on recapitulating CODA phenotypes in animal models and assessing the mechanism of MMP19 involvement during development.

Orientadores: Mônica Barbosa de Melo, José Paulo Cabral de Vasconcellos / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-25T15:53:03Z (GMT). No. of bitstreams: 1 Oliveira_MarianaBorges_M.pdf: 6901699 bytes, checksum: b84b4ac92949fdb082625b01ad6c66a1 (MD5) Previous issue date: 2014 / Resumo: O glaucoma engloba um grupo de doenças que têm como característica comum a atrofia progressiva do disco óptico com alteração correspondente de campo visual, decorrente da perda de células ganglionares da retina. O aumento da pressão intraocular (PIO) é o principal fator de risco para o desenvolvimento do glaucoma, mas tem sido sugerida a existência de outros aspectos relevantes, tais como alterações no metabolismo do óxido nítrico, na regulação do fluxo vascular, sinais de estresse oxidativo e alterações no sistema imunológico. O glaucoma primário de ângulo aberto (GPAA), a forma mais prevalente entre os glaucomas, está associado a uma série de fatores de risco para sua instalação e desenvolvimento, além da PIO. Os demais fatores de risco são: idade (relação direta), raça (mais frequente e grave em indivíduos da raça negra), miopia e história familiar de glaucoma. A maioria dos casos de GPAA apresenta um padrão complexo de hereditariedade, em que variantes de suscetibilidade contribuem para a gravidade da doença. O objetivo deste estudo do tipo caso-controle foi avaliar o papel de polimorfismos nos genes da interleucina alfa (IL1A), interleucina beta (IL1B) e fator de necrose tumoral alfa (TNFA) em relação à suscetibilidade ao GPAA. Após avaliação oftalmológica, foram recrutados 214 indivíduos não relacionados portadores de GPAA. A análise molecular foi realizada por meio de técnicas de PCR e sequenciamento direto. Neste estudo observou-se que o alelo C do polimorfismo -31 C/T, presente na região promotora do gene IL1B, confere risco para o glaucoma (p=0,002). Este alelo está contido em um "TATA" box, influenciando na transcrição deste gene. Existe um desequilíbrio de ligação entre os SNPs -31 C/T e -511 T/C do gene IL1B e este último também está associado a um risco aumentado para o glaucoma (p=0,006). Com relação aos demais SNPs estudados (+3954 C/T do gene IL1B, -889 C/T e +4845 G/T do gene IL1A e -238 G/A e -308 G/A do gene TNFA), não houve associação com o glaucoma. Este estudo sugere que há relação das variantes "C" do SNP -31 C/T e "T" do SNP -511 T/C do gene IL1B com a etiologia do GPAA em uma amostra da população brasileira / Abstract: Glaucoma comprises a group of diseases characterized by progressive atrophy of the optic disc and corresponding visual field loss, as consequence of retinal ganglion cells death. Augmented intraocular pressure (IOP) is the main risk factor for glaucoma development, but it has been suggested that there are other relevant aspects that may lead to neuronal and trabecular meshwork (TM) damage and contribute to the development of glaucoma, including alterations in nitric oxide metabolism, oxidative stress and vascular and immune system dysregulations. Primary open angle glaucoma (POAG), the most prevalent form of glaucoma, is associated with several risk factors to its initiation and progression besides IOP. The additional risk factors are: age (direct correlation), race (more frequent in black subjects), myopia and family history of glaucoma. The majority of POAG cases presents a complex pattern of heritability, in which susceptibility variants contribute to disease severity. The aim of this case-control study was to evaluate the role of single nucleotide polymorphisms (SNPs) in the interleukin alpha (IL1A), interleukin beta (IL1B) and tumor necrosis factor alpha (TNFA) genes in relation to POAG susceptibility. After ophthalmologic evaluation, 214 unrelated subjects with POAG were recruited. The molecular analysis was performed by PCR and direct sequencing techniques. In this study it was observed that the C allele of the -31 C/T promoter polymorphism of the IL1B gene confers risk for glaucoma (p=0.002). This allele disrupts a "TATA" box, influencing the transcription of the IL1B gene. There is linkage disequilibrium between the -31 C/T e -511 T/C SNPs, and the latter is also associated with an increased risk for glaucoma (p=0.006). Regarding the others studied SNPs (+3954 C/T of the IL1B gene, -889 C/T and +4845 G/T of the IL1A gene and -238 G/A and -308 G/A of the TNFA gene), there is no association with glaucoma. This study suggests the relationship of the "C" variant of -31C/T and "T" variant of -511T/C of IL1B gene with POAG etiology in a sample of the Brazilian population / Mestrado / Oftalmologia / Mestra em Ciências Médicas

Glaucoma is the third most common cause of blindness in the world. There are several types of glaucomma, which makes early diagnosis of this illness harder. One posibble way for early diagnosis could be analysis of a retinal vessel pulsation. Data in this work were captured by experimental device called video ophtalmoscope. Several methods for optic disc segmentation were designed. From segmented parts, pulsation curve was recorded. Analysis of the pulsation curve was provided in two ways: Analysis of the whole pulsation curve and Averaged pulse analysis. Both methods were tested with diferent reference signals.

Title: Gradient boosted segmentation of retinal fundus images Author: Matúš Goliaš Department: Department of Software and Computer Science Education Supervisor: Doc. RNDr. Elena Šikudová PhD., Department of Software and Computer Science Education Abstract: Over the recent years, there has been an increase in the use of automatic methods in medical diagnosis. A significant number of publications have analysed eye disorders and diseases. One of the most severe eye conditions is glaucoma. It damages optic nerves and causes gradual loss of vision. An essential step towards a faster diagnosis of this disease is accurate segmentation of the optic disc and cup. This task is difficult due to many retinal defects, different image acquisition techniques, and artefacts caused by imaging devices. This thesis describes an iterative threshold-based algorithm for extraction of the optic disc. An objective function quantifying object similarity to the optic disc is defined to direct the iteration. Following that, we introduce a superpixel-based classification algorithm for extraction of the optic cup. We propose the use of gradient boosted decision trees which outperform random forest and support vector machine. In addition, we evaluate the proposed algorithms and their alternatives on a publicly available retinal fundus...

Amblyopia is the leading cause of monocular visual impairment in children. Therapy for amblyopia is extremely beneficial in some children but ineffective in others. It is critical that the reasons for this discrepancy are understood. Emerging evidence indicates that current clinical protocols for the diagnosis of amblyopia may not be sufficiently sensitive in identifying individuals who, on more detailed examination, exhibit subtle structural defects of the eye. Presently, the magnitude of this problem is unknown. The aim of this study was to establish the prevalence of subtle retinal/optic nerve head defects in eyes diagnosed with amblyopia, to distinguish between possible explanations for the origin of such defects and to investigate the relationship between quantitative measures of retinal structure, retinal nerve fibre layer thickness and optic nerve head dimensions. Using the imaging technique of Optical Coherence Tomography (OCT) retinal structure has been investigated in detail, following the visual pathway across the retina from the fovea, via the paramacular bundle to the optic disc, where peripapillary retinal nerve fibre thickness has been imaged and subjected to detailed measures along with optic disc size and shape. The study formed two phases, the first imaging the eyes of visually normal adults and children, comparing them to amblyopes, both adults and children who had completed their treatment. The second phase, a longitudinal study, investigated retinal structure of amblyopic children undertaking occlusion therapy for the first time. By relating pre-therapy quantitative measures to the visual outcome the second phase of the study aimed to examine whether OCT imaging could identify children achieving a poor final outcome. The results show a clear picture of inter-ocular symmetry structure in all individuals, visually normal and amblyopic. Optic disc characteristics revealed no structural abnormalities in amblyopes, in any of the measured parameters, nor was there any association between the level of visual acuity and the measured structure. At the fovea differences were shown to occur in the presence of amblyopia, with thickening of the fovea and reduction of the foveal pit depth. The structural changes were found to be both bilateral and symmetrical with the fellow eye also affected. In the longitudinal phase of the study these changes were demonstrated to a greater extent in children who ¿failed¿ to respond to treatment. This bilateral, symmetrical structural change found at the fovea, which has not been previously reported, cannot therefore be the primary cause of the visual loss which has been diagnosed as amblyopia.

The optic disc is the region of the retina where the optic nerve exits the back of the eye. A number of conditions can cause the optic disc to swell. Papilledema, optic disc swelling caused by raised intracranial pressure (ICP), and nonarteritic anterior ischemic optic neuropathy (NAION), swelling caused by reduced blood flow to the back of the eye, are two such conditions. Rapid, accurate diagnosis of the cause of disc swelling is important, as with papilledema the underlying cause of raised ICP could potentially be life-threatening and may require immediate intervention. The current clinical standard for diagnosing and assessing papilledema is a subjective measure based on qualitative inferences drawn from fundus images. Even with the expert training required to properly perform the assessment, measurements and results can vary significantly between clinicians. As such, the need for a rapid, accurate diagnostic tool for optic disc swelling is clear. Shape analysis of the structures of the retina has emerged as a promising quantitative tool for distinguishing between causes of optic disc swelling. Optic disc swelling can cause the retinal surfaces to distort, taking on shapes that differ from their normal arrangement. Recent work has examined how changes in the shape of one of these surfaces, Bruch's membrane (BM), varies between different types of optic disc swelling, containing clinically-relevant information. The inner limiting membrane (ILM), the most anterior retinal surface and furthest from BM, can take on shapes that are distinct from the more posterior layers when the optic disc becomes swollen. These unique shape characteristics have yet to be explored for their potential clinical utility. This thesis develops new shape models of the ILM. The ultimate goal of this work is to develop noninvasive, automated diagnostic tools for clinical use. To that end, a necessary first step in establishing clinical relevance is demonstrating the utility of retinal shape information in a machine learning classifier. Retinal layer shape information and regional volume measurements acquired from spectral-domain optical coherence tomography scans from 78 patients (39 papilledema, 39 NAION) was used to train random forest classifiers to distinguish between cases of papilledema and NAION. On average, the classifiers were able to correctly distinguish between papilledema and NAION 85.7±2.0% of the time, confirming the usefulness of retinal layer shapes for determining the cause of optic disc swelling. The results of this experiment are encouraging for future studies that will include more patients and attempt to differentiate between additional causes of optic disc edema.

Optical coherence tomography (OCT) is a safe and non-invasive imaging technique providing high axial resolution. A spectral-domain OCT scanner capable of acquiring volumetric data of the retina is becoming an increasingly important modality in ophthalmology for the diagnosis and management of a variety of retinal diseases such as glaucoma, diabetic retinopathy and age related macular degeneration (AMD) which are major causes of a loss of vision. To analyze and track these ocular diseases, developments of the automated methods for detecting intraretinal layers, optic discs and retinal blood vessels from spectral-domain OCT scans are highly required recently. The major contributions of this thesis include: 1) developing a fast method that can automatically segment ten intraretinal layers in the spectral-domain macular OCT scan for the layer thickness analysis, 2) developing a method that can automatically segment the optic disc cup and neuroretinal rim in the spectral-domain OCT scan centered at the optic nerve head (ONH) to measure the cup-to-disc ratio, an important structural indicator for the progression of glaucoma, and 3) developing a method that can automatically segment the 3-D retinal blood vessels in the spectral-domain ONH-centered OCT scan to extract 3-D features of the vessels for the diagnosis of retinal vascular diseases.

This thesis deals with the analysis of retinal images from digital fundus camera, especially with structure of optic disc (OD). The theoretic part describes main features of the human visual system and princip of eyeground examination. The paper discussed available methods that have been used for localization of optic disc. For further work is selected proposal of the geometrical model of vessel structure by using the method of segmentation of vessels by matched filtering. Original images are tested with semi-automatic method for optic disc localization.

This thesis deals with the analysis of retinal images from digital fundus camera, especially with structure of optic disc (OD). The theoretic part describes main features of the human visual system and princip of eyeground examination. The paper discussed sevaral available methods that have been used for localization of the optic disc. In the second part is proposed the automatic detector of the optic disc based on the principle of genetic algorithm by using the method of segmentation of vessels by matched filtering. Original images are tested with this automatic method.

Approximate Nearest-Neighbour Field (ANNF\ maps between two related images are commonly used by computer vision and graphics community for image editing, completion, retargetting and denoising. In this work we generalize ANNF computation to unrelated image pairs. For accurate ANNF map computation we propose Feature Match, in which the low-dimensional features approximate image patches along with global colour adaptation. Unlike existing approaches, the proposed algorithm does not assume any relation between image pairs and thus generalises ANNF maps to any unrelated image pairs. This generalization enables ANNF approach to handle a wider range of vision applications more efficiently. The following is a brief description of the applications developed using the proposed Feature Match framework. The first application addresses the problem of detecting the optic disk from retinal images. The combination of ANNF maps and salient properties of optic disks leads to an efficient optic disk detector that does not require tedious training or parameter tuning. The proposed approach is evaluated on many publicly available datasets and an average detection accuracy of 99% is achieved with computation time of 0.2s per image. The second application aims to super-resolve a given synthetic image using a single source image as dictionary, avoiding the expensive training involved in conventional approaches. In the third application, we make use of ANNF maps to accurately propagate labels across video for segmenting video objects. The proposed approach outperforms the state-of-the-art on the widely used benchmark SegTrack dataset. In the fourth application, ANNF maps obtained between two consecutive frames of video are enhanced for estimating sub-pixel accurate optical flow, a critical step in many vision applications. Finally a summary of the framework for various possible applications like image encryption, scene segmentation etc. is provided.