O albinismo e uma alteracao genetica rara que compromete a producao de melanina. As alteracoes clinicas consistem na falta de pigmentacao da pele, cabelo e pelos. Apresenta alteracoes oftalmologicas importantes, que comprometem a acuidade visual de forma severa, na grande maioria dos casos. As alteracoes oftalmologicas sao, ametropias, nistagmo, rarefacao do epitelio pigmentado da iris e da retina, hipoplasia foveal e decussacao anomala do nervo optico. Este estudo foi dividido em tres diferentes partes com os seguintes objetivos principais: Parte 1: Analise comparativa entre o grau de transparencia da iris (GTI), o grau de transparencia da retina (GTR) e a espessura da regiao macular com a acuidade visual nos pacientes com albinismo. Parte 2: Teste de sensibilidade ao contraste espacial de luminancia e ofuscamento com lentes de contato filtrantes em ambientes claros e escuros. Parte 3: Avaliacao da visao de cores, utilizando\\se o teste de Pranchas de Ishihara e o Cambridge Color Test (CCT). Material\' e\' Métodos: Participaram do estudo 121 individuos albinos, com idade media de 31,18 (} 15,47) anos, o que totalizou 242 olhos. Os participantes foram divididos em diferentes grupos nas tres partes do estudo, alguns participaram de uma ou mais partes. Na primeira parte os participantes foram submetidos a consulta oftalmologica, classificacao do grau de transparencia da iris e da retina em uma nova classificacao baseada em quatro diferentes graus de transparencia, alem de avaliacao foveal pelo exame de Tomografia de Coerencia Optica (OCT). Na segunda parte, os participantes foram submetidos ao teste de contraste computadorizado, utilizando lentes de contato filtrante e transparentes, em ambientes claros e escuros. Nesta etapa foi avaliada a influencia da lente filtrante na sensibilidade de contraste visual dos individuos com albinismo. Na terceira parte, os participantes foram submetidos aos testes de Pranchas de Ishihara e de Cambridge Colour Test (CCT), para avaliacao da visao de cores nos pacientes albinos. Resultados: Os resultados foram os seguintes: Parte\'1: A correlacao entre AV logMAR e GTI foi positiva (+0,569) e significativa (p<0,001). A correlacao entre AV logMAR e GTR foi positiva (+0,531) e significativa (p<0,001). A correlacao entre AV logMAR e a espessura macular nao foi significativa (p=0,105). A correlacao entre GTI e GTR foi positiva (+ 0,627) e significativa (p<0,001). A correlacao entre espessura macular e GTI nao foi significativa (p=0,397). A correlacao entre espessura macular e GTR nao foi significativa (p=0,458). Parte 2: Houve melhora estatisticamente significante da sensibilidade ao contraste com as lentes filtrantes em relacao as lentes transparentes no ambiente de ofuscamento (claro), na frequencia de 0,3 cpg, 0,6 cpg e 1,0 cpg (ciclos por grau). Nao houve melhora significante nas frequencia 2,0 cpg e 4,0 cpg. Nao houve melhora estatisticamente significante da sensibilidade ao contraste de lentes filtrantes em relacao as lentes transparentes no ambiente escuro (penumbra) em nenhuma das frequencias espaciais estudadas. Parte\' 3: Todos os individuos com albinismo nao apresentaram nenhuma alteracao de visao de cores no teste de Pranchas de Ishihara. O CCT demonstrou piora no limiar de deteccao de cores significativa nos eixos protan (p=0,021) e deutan (p=0,017), mas nao houve diferenca no eixo tritan (p=0,212). Os testes estatisticos sugerem que uma amostra maior seja estudada para validacao dos resultados / Albinism is a rare genetic alteration that compromises the production of melanine in all body tissues. The clinical alterations are the lack of pigment in the skin and hair. It presents important ophthalmological changes that compromise the visual acuity, in most cases. The ophthalmological changes are: ametropia, nystagmus, iris pigmented epithelium rarefaction, retina pigmented epithelium rarefaction, foveal hipoplasia and abnormal optic nerve decussation. This study has been divided in three different parts and had the following main objectives: Part 1, comparative analysis of the iris transparency degree (ITD), the retina transparency degree (RTD) and macular thickness with the visual acuity in albino patients. Part 2: Spatial contrast sensitivity test of luminance and outshine wearing contact lenses with filters in light and dark environments. Part 3: color vision evaluation using the Ishihara Test and the Cambridge Color Test (CCT). For the study 121 individuals with albinism were included with an average of 31,18 years old } 15,47, a total of 242 eyes. The participants were divided in different groups in the three parts of the study, some participated in one or more parts. In the first part the participants have been through an ophthalmological exam, classification of the iris transparency degree (ITD) and the retinal transparency degree (RTD) by a new classification based in 4 different degrees of transparency. Foveal evaluation by the Optic Coherence Tomography exam (OCT) was made in the first part of this study. In the second part, the participants have been submitted to the computerized contrast test, using filtered and transparent contact lenses in light and dark environments. In this part the influence of the filtering lenses in the visual contrast sensitivity in the albinism patients has been evaluated. In the third part, the participants have been submitted to Ishihara Test and Cambridge Color Test in order to evaluate the color vision in albino patients. The results are as follows: Part\' 1: the correlation between visual acuity in Logarithm of the Minimum Angle of Resolution (logMAR) and ITD was positive (+0,569) and significant (p<0,001). The correlation between visual acuity in logMAR and RTD was positive (+0,531) and significant (p<0,001). The correlation between visual acuity in logMAR and the macular thickness was not significant (p=0,105). The correlation between ITD and RTD was positive (+ 0,627) and significant (p<0,001). The correlation between the macular thickness and the ITD was not significant (p=0,397). The correlation between the macular thickness and the RTD was not significant (p=0,458). Part\'2: There has been a statistically significant improvement of the contrast sensitivity wearing the filtering lenses in relation to the transparent ones in bright environment, in the frequencies of 0,3 cycles per degree (cpd), 0,6 cpd and 1,0 cpd. There has been no significant improvement in the frequencies 2,0 cpd and 4,0 cpd. There has been no statistically significant improvement in contrast sensitivity wearing the filtering lenses in relation to the transparent one in the dark environment (shadow) in any of the spatial frequencies studied. Part\' 3: all albinism patients did not present any color vision change in the Ishihara Test. The CCT has shown a worsening in the color detection threshold in the protan axis (p=0,021) and deutan axis (p=0,017), but there has been no difference in the tritan axis (p=0,212). The statistic tests show that a bigger sample is recommended to confirm part 3 results

No / Retinal dystrophies are an overlapping group of genetically heterogeneous conditions resulting from mutations in more than 250 genes. Here we describe five families affected by an adult-onset retinal dystrophy with early macular involvement and associated central visual loss in the third or fourth decade of life. Affected individuals were found to harbor disease-causing variants in DRAM2 (DNA-damage regulated autophagy modulator protein 2). Homozygosity mapping and exome sequencing in a large, consanguineous British family of Pakistani origin revealed a homozygous frameshift variant (c.140delG [p.Gly47Valfs∗3]) in nine affected family members. Sanger sequencing of DRAM2 in 322 unrelated probands with retinal dystrophy revealed one European subject with compound heterozygous DRAM2 changes (c.494G>A [p.Trp165∗] and c.131G>A [p.Ser44Asn]). Inspection of previously generated exome sequencing data in unsolved retinal dystrophy cases identified a homozygous variant in an individual of Indian origin (c.64_66del [p.Ala22del]). Independently, a gene-based case-control association study was conducted via an exome sequencing dataset of 18 phenotypically similar case subjects and 1,917 control subjects. Using a recessive model and a binomial test for rare, presumed biallelic, variants, we found DRAM2 to be the most statistically enriched gene; one subject was a homozygote (c.362A>T [p.His121Leu]) and another a compound heterozygote (c.79T>C [p.Tyr27His] and c.217_225del [p.Val73_Tyr75del]). DRAM2 encodes a transmembrane lysosomal protein thought to play a role in the initiation of autophagy. Immunohistochemical analysis showed DRAM2 localization to photoreceptor inner segments and to the apical surface of retinal pigment epithelial cells where it might be involved in the process of photoreceptor renewal and recycling to preserve visual function.

Åldersrelaterad makuladegeneration (AMD) är en kronisk ögonsjukdom och är den vanligaste orsaken till irreversibel synnedsättning hos äldre personer i den industriella världen. Bakomliggande orsaker kan vara både genetiska och miljömässiga varför prevention och behandling av ögonsjukdomar kan öka livskvaliteten samt minska nedsatt syn och de funktionshinder som synnedsättning medför. Syftet med studien var att belysa ögonsjuksköterskans hälsofrämjande arbete med patienter som har åldersrelaterad makuladegeneration. Denna integrativa litteraturstudie visade att kunskapsnivån om AMD var låg både hos den allmänna befolkningen och patienter. Även patienter med allvarlig synnedsättning på grund av en eller flera ögonsjukdomar var omedvetna om sin diagnos. Patienter med AMD upplevde att de inte fick tillräckligt med information om sjukdomen, riskfaktorer, undersökningar och vilka behandlingar som fanns. Att drabbas av AMD innebar en stor påverkan på patienternas livskvalitet. Det visade sig att det fanns ett starkt samband mellan förlust av synfunktionen och symtom på depression. Ingen av studierna var från Sverige eller belyste svenska förhållanden men belyser vikten av att hälsofrämjande arbete och ökad medvetenhet kring kroniska ögonsjukdomar och dess riskfaktorer är mycket viktigt. Det vore av stort intresse att undersöka de svenska förhållandena som jämförelse. / Age-related macular degeneration (AMD) is a chronic eye disease and is the most common cause of irreversible vision loss amongst elderly people within the industrial world. Underlying causes can be both genetical and environmental, prevention and treatment of AMD can increase quality of life and reduce visual impairment and all dysfunctionalities related to it. The aim of this study was to highlight ophthalmic nurses' health promoting work with patients with age-related macular degeneration. This integrative literature review showed that the level of awareness of AMD was low among both the general population and patients. Even patients with severe vision loss due to one or several eye diseases was unaware of their diagnosis. Patients with AMD felt that they were not given enough information about the disease, the risk factors, available health examinations and what possible treatments were available. To be affected with AMD entails a significant negative effect on patients quality of life. It was shown that there was a strong connection between vision loss and symptoms of depression. None of the studies were from Sweden or highlighted the Swedish conditions but highlights the importance of health promotion, awareness about chronic eye disease and its risk factors as very important. It would be of great interest to investigate the Swedish conditions for comparison.

The plasma protein complement factor H (fH, 155 kDa) regulates the activity of the alternative pathway of complement activation. Factor H is monomeric, and its 20 CCP modules are arranged in a predominantly elongated conformation, joined by linking sequences that vary in length, with the longest linkers occurring in the central portion of the molecule. CCP modules 1 through 4 of fH host its capacity to act as a cofactor for fI-mediated proteolytic degradation of C3b and its ability to accelerate the decay of the C3 convertase, C3bBb, thereby regulating the so-called tick-over activation of the alternative pathway. Mutations in this part of fH might compromise its function and lead to underregulation of the alternative pathway. It is hypothesized that this can cause predisposition to diseases such as atypical haemolytic uraemic syndrome (aHUS) and age-related macular degeneration (AMD). In the current work, the known disease-associated mutations R53H and R78G were compared to wild-type in terms of fluid-phase cofactor assays, C3b-binding affinity and the ability to accelerate the decay of the convertase. In addition, the protective variant, I62, was also inspected because its protective role might be explained by an increased regulatory activity. The second, linked, aim of this project was to employ Forster resonance energy transfer (FRET) to study the link between conformation and function in fH. FRET is valuable for obtaining long-distance restraints up to a maximum of 100 °A and is therefore particularly useful for inferring domain orientations within multidomain proteins. This approach to measure long-range inter- and intramolecular distances is a convenient way to complement NMR-based structural investigations, which rely on short-range restraints. It is also a valuable complement to X-ray crystallography since it is a solution technique that can be conducted under physiological conditions. By using site-directed mutagenesis in the current work, free cysteines were introduced into CCP modules 1-4 at strategic points, which were then used for attachment of fluorescent tags. C3 possesses an internal thioester which can be labelled with a fluorophore upon activation to C3b. Intermolecular FRET measurements were thus undertaken to gain information about the interaction between the two proteins that is crucial for understanding functional activity. The CCP modules in the centre of fH may be responsible for introducing a bend into fH that brings the N-teminus close to the C-terminus (the latter is important for host versus non-host discrimination) joined by the longest linkers occurring in the whole molecule. This coincidence of two relatively small CCP modules, 12 and 13, with the highest number of eight amino acids between them, is hypothesised to reflect some unique architectural features. To explore the structural details of this portion of fH by FRET, single-labelled cysteine mutants were further modifed to provide a recognition site for transglutaminase (TGase), which can be enzymatically labeled with a second fluorophore. This stoichiometrically-labelled protein was used for intramolecular FRET studies.

The aims of this study were to analyse known and potential biomarkers of common and genetically complex human disorders and to identify genetic and environmental variation associated with plasma biomarker concentrations. Two groups of protein biomarkers were analysed. First, plasma complement factor H (CFH) was selected as a potential biomarker for age-related macular degeneration (AMD), since common variants in the CFH gene show strong association with this disorder. Secondly, two isoforms of amyloid-β (Aβ40 and Aβ42) were selected as biomarkers for Alzheimer disease (AD) since Aβ deposits are major constituents of the amyloid plaques characteristic of this disorder. Physiological and anthropometric measurements and samples of human and genomic DNA were collected from a population sample of 1,021 individuals from the Croatian island of Vis. Quantitative determination of plasma Aβ40 and Aβ42 concentrations was performed using enzyme-linked immunosorbent assays. Heritabilities and significant covariate effects were estimated for each trait in the Croatian data set. Genome-wide linkage and association analyses were conducted for the biomarker traits. A novel finding was the genome-wide significant association between a CFH and several polymorphisms close to and within the CFH gene. The strongest association was with an intronic SNP within CFH, which explained 28% of the total trait variance (P < 10-50). The association was also replicated in a Dutch sample set. A SNP haplotype was identified which accounted for a higher proportion of the phenotypic variance. Conditional haplotype analysis showed that the effect of this haplotype on plasma CFH concentration was independent of the CFH Y402H variant, and significantly stronger than a deletion of the adjacent CFHR3/CFHR1 which was already known to affect AMD susceptibility. Genetic analysis of 382 AMD cases and 201 controls was consistent with the CFH Y402H variant being the strongest AMD susceptibility locus. Variation in plasma CFH concentration was found to explain up to 1.8% of the variation in susceptibility to AMD with an odds 2.1 (95% C.I. 1.3-3.4, P = 0.003). SNPs that were strongly associated with a CFH concentration also influenced AMD susceptibility (P < 0.05) independently of the CFH Y402H polymorphism. Functional analysis of genomic regions associated with plasma CFH is needed to identify the causal variants. Associations were observed between plasma Aβ40 concentration and several novel candidate loci, spanning regions of approximately 0.2 Mb, on chromosomes 9 and X. Similarly, novel associations with plasma Aβ42 were found in several regions, each spanning 0.2-0.4 Mb, on chromosomes 2, 5, 9, 15 and 20. The proportion of the phenotypic variance in plasma Aβ42 explained by these putative associations ranged between 1.8 and 2.8%. However, none of the associated SNPs was significant after correction for multiple testing, therefore replication is required. Finally, attempts were made to identify and quantitate new protein biomarkers of disease in human plasma using mass spectrometry. Development and optimisation of techniques was initially undertaken to deplete high-abundance plasma proteins and improve signal:noise ratio. This allowed the assessment of downstream proteomic approaches including MALDI-TOF mass spectrometry (MS), capillary electrophoresis (CE) and ion exchange chromatography (IEC), each with the potential for large-scale quantitation of plasma proteins. Although the analysis of single protein analytes, using CE and IEC proved promising, the results highlighted the difficulty associated with MALDI-TOF and protein ionisation techniques in analysing complex mixtures such as plasma.

In patients with age-related macular degeneration, a monolayer of cells in the eyes called retinal pigment epithelium differ from healthy ones in morphology. It is therefore important to quantify the morphological changes, which will help us better understand the physiology, disease progression and classification. Classification of the RPE morphometry has been accomplished with whole tissue data. In this work, we focused on the spatial aspect of RPE morphometric analysis. We used the second-order spatial analysis to reveal the distinct patterns of cell clustering between normal and diseased eyes for both simulated and experimental human RPE data. We classified the mouse genotype and age by the k-Nearest Neighbors algorithm. Radially aligned regions showed different classification power for several cell shape variables. Our proposed methods provide a useful addition to classification and prognosis of eye disease noninvasively.

A Thesis submitted to The University of Arizona College of Medicine - Phoenix in partial fulfillment of the requirements for the Degree of Doctor of Medicine. / Genetic association studies simplify genotype‐phenotype relationship investigation by considering only the presence of a given polymorphism and the presence or absence of a given downstream phenotype. Although such associations do not indicate causation, collections of phenotypes sharing association with a single genetic polymorphism may provide valuable mechanistic insights. In this thesis we explore such genetic pleiotropy with Deep Phenotype Association Studies (DeePAS) using data from the Age‐Related Eye Study 2 (AREDS2). We also employ a novel text mining approach to extract pleiotropic associations from the published literature as a hypothesis generation mechanism. Is it possible to identify pleiotropic genetic associations across multiple published abstracts and validate these in data from AREDS2? Data from the AREDS2 trial includes 123 phenotypes including AMD features, other ocular conditions, cognitive function and cardiovascular, neurological, gastrointestinal and endocrine disease. A previously validated relationship extraction algorithm was used to isolate descriptions of genetic associations with these phenotypes in MEDLINE abstracts. Results were filtered to exclude negated findings and normalize variant mentions. Genotype data was available for 1826 AREDS2 participants. A DeePAS was performed by evaluating the association between selected SNPs and all available phenotypes. Associations that remained significant after Bonferroni‐correction were replicated in AREDS. LitWAS analysis identified 9372 SNPs with literature support for at least two distinct phenotypes, with an average of 3.1 phenotypes/SNP. PheWAS analyses revealed that two variants of the ARMS2‐HTRA1 locus at 10q26, rs10490924 and rs3750846, were significantly associated with sub‐retinal hemorrhage in AMD (rs3750846 OR 1.79 (1.41‐2.27), p=1.17*10‐7). This associated remained significant even in populations of participants with neovascular AMD. Furthermore, odds ratios for the development of sub‐retinal hemorrhage in the presence of the rs3750846 SNP were similar between incident and prevalent AREDS2 sub‐populations (OR: 1.94 vs 1.75). This association was also replicated in data from the AREDS trial. No literature‐defined pleiotropic associations tested remained significant after multiple‐testing correction. The rs3750846 variant of the ARMS2‐HTRA1 locus is associated with sub‐retinal hemorrhage. Automatic literature mining, when paired with clinical data, is a promising method for exploring genotype‐phenotype relationships.

La rétinopathie diabétique (RD) se compose d’une part d’une ischémie rétinienne périphérique et d’autre part d’une exsudation rétinienne responsable d’un œdème maculaire diabétique, première cause de cécité chez les moins 55 ans. Les traitements utilisés actuellement sont non spécifiques et traitent les complications tardives de la RD. Les phases précoces de la RD ne sont donc pas ciblées. L’hyperglycémie chronique entraine un stress oxydant et une activation des PKC qui participent à l’altération des BHR. L’objectif de ce travail a été 1°) d’étudier l’implication de la PKCζ et de la voie Rho/ROCK/Myosine II sur la physiopathogénie de la RD et 2°) de montrer l’effet bénéfique de leurs inhibiteur sur les BHR et sur la reperfusion des capillaires rétiniens. Nous avons confirmé l’hyperactivation de la PKCζ et de la voie Rho/ROCK/Myosine II chez les rats diabétiques et leur participation à la rupture de la BHR externe. Le traitement par leurs inhibiteurs respectifs normalise l’activation des deux enzymes et restaure l’intégrité anatomique et fonctionnelle de la BHR externe. De plus l'hyperactivation de ROCK altère la perfusion rétinienne par 1) constriction focale artériolaire, 2) protrusions membranaires endoluminales des cellules endothéliales (blebbing) et 3) vasoconstriction capillaire diffuse. Nous avons montré que l'ensemble de ces phénomènes étaient réversibles par traitement intravitréen de son inhibiteur le Fasudil. De manière importante le traitement par Fasudil induit également une diminution du VEGF rétinien responsable de la perméabilité des barrières et témoin indirect de l’ischémie rétinienne. Ces travaux éclairent la physiopathogénie de la RD et ouvre des perspectives thérapeutiques permettant de cibler les événements précoces de la RD. / Diabetic retinopathy (DR) mainly results from peripheral retinal ischemia and exudation leading to sight threatening complications such as retinal neovascularization or macular edema. This latter represents the main cause of visual loss among working age individuals. Current treatments address late complications of DR and are non-specific. Therefore, early events are currently not addressed. Chronic hyperglycemia increases oxidative stress and activates PKC leading to blood retinal barrier (BRB) breakdown. The aims of the present work were two fold. First, to assess the implication of PKCζ and the Rho/ROCK/Myosin II pathway on the pathogenesis of DR and second, to assess whether their specific inhibitors have the potential to restore the phenotype. Herein we have demonstrated the pathogenic role of PCKζ and ROCK hyperactivation on the development of diabetes induced external BRB breakdown. Furthermore their inhibitors restored the morphologic and functional aspect of the external BRB. We also found that ROCK hyperactivation was responsible for altered retinal perfusion through several mechanism namely 1) focal constriction of retinal arterioles; 2) endoluminal protrusions of the endothelial cell membrane (blebs) and 3) capillary diffuse vasoconstriction. We were able to demonstrate that all this aspects were reversible by Fasudil, a ROCK inhibitor, administrated into the vitreous. Of importance this treatment decreased also retinal VEGF that is a well-known factor responsible for barrier breakdown and a surrogate marker for retinal ischemia. To conclude the present findings not only shed light on the mechanisms of DR but also open new therapeutic avenues addressing the early events of DR a current unmet medical need.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-03-22T12:41:44Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_VariabilidadeAlelicaExpressao.pdf: 6674673 bytes, checksum: 0ec2d9f9a77ab69b8cf5669cccae4516 (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-03-27T11:34:28Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_VariabilidadeAlelicaExpressao.pdf: 6674673 bytes, checksum: 0ec2d9f9a77ab69b8cf5669cccae4516 (MD5) / Made available in DSpace on 2017-03-27T11:34:28Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_VariabilidadeAlelicaExpressao.pdf: 6674673 bytes, checksum: 0ec2d9f9a77ab69b8cf5669cccae4516 (MD5) Previous issue date: 2015-06-24 / A maculopatia autossômica recessiva de Stargardt se caracteriza pela perda progressiva simétrica da visão na região central do campo visual em indivíduos entre a primeira e segunda décadas de vida. Também fazem parte do diagnóstico clínico a presença de lesões maculares pontuais de coloração amarelo-esbranquiçadas encontradas nas regiões foveal e parafoveal, além da deposição de lipofuscina gerando hipofluorescência e atrofia córiocapilar e do epitélio pigmentado da retina. A acuidade visual dos portadores da degeneração de Stargardt decresce com o seu progresso e tende a se estabilizar entre 20/200 (1,0 logMAR) a 20/400 (1,3 logMAR). Esta tese teve como objetivos a investigação da ocorrência de associações entre as variantes alélicas (mutações) e os níveis de expressão do gene ABCA4 com a morfologia dos granulócitos periféricos e os fenótipos clínicos, eletrofisiológicos e de estrutura da retina em pacientes portadores da maculopatia de Stargardt. Nos onze sujeitos selecionados foram identificadas quatro variantes do tipo não-sinônimas e três do tipo sinônimas. A combinação alélica L1395P/D1817E foi encontrada em 64% dos 22 cromossomos analisados sugerindo a ocorrência do efeito do fundador. Não foi possível a associação entre os genótipos e os fenótipos analisados. Os valores diferenciais de expressão do gene ABCA4 obtidos entre pacientes e grupo controle, bem como a prevalência de neutrófilos bastonados na circulação sanguínea periférica sugerem a possibilidade de servirem como biomarcadores para Stargardt. As imagens retinográficas e angiográficas obtidas permitiram a classificação em estágios I, II e III de comprometimento retiniano nos sujeitos investigados. O valores de espessura da região central da mácula dos pacientes foram bem menores do que aqueles obtidos para os controles, evidenciando a perda da camada de fotorreceptores. Os achados eletrorretinográficos de campo total em função do tempo de adaptação ao claro possibilitaram a caracterização das perdas funcionais nos sujeitos investigados. / The autosomal recessive Stargardt's maculopathy is characterized by symmetrical progressive loss of central vision in patients’ first and second decades of life. It is also part of the clinical diagnosis the presence of yellowish-white color specific macular lesions found in the parafoveal and foveal regions, besides lipofuscin deposition generating hypofluorescence and atrophy of choriocapillaris and retinal pigment epithelium. Visual acuity of the Stargardt’s degeneration carriers decreases with their progress and tend to stabilize between 20/200 (1.0 logMAR) to 20/400 (1.3 logMAR). This thesis aimed to investigate the occurrence of association between allelic variants (mutations) and ABCA4 gene expression levels with the peripheral granulocytes morphology and clinical phenotypes, electrophysiological and retinal structure in patients harboring Stargardt’s maculopathy. Four non-synonymous and three synonymous allelic variants were identified among eleven selected subjects. The allelic combination L1395P/D1817E was found in 64% of the 22 chromosomes analyzed suggesting the occurrence of founder effect. There were no association between genotypes and clinical phenotypes analyzed. The differential values obtained for ABCA4 gene expression between patients and controls, as well as the prevalence of banded neutrophils in the peripheral blood circulation suggest the possibility they could serve as biomarkers for Stargardt. The eye fundus and angiographic images were used for the classification in the stages I, II and III of retinal impairment in subjects investigated. The thickness values of the central region of the macula among patients were much lower than those obtained for the controls, indicating a loss of photoreceptor layer. Full-field electroretinographic findings of light adapted eyes during the course of time enabled the characterization of the functional losses in investigated subjects.

Dissertação de Mestrado em Nutrição Clínica apresentada à Faculdade de Ciências da Nutrição e Alimentação da Universidade do Porto / Resumo da dissertação:Introdução: A degenerescência macular da idade (DMI) é a principal causa de perda visual grave acima dos 50 anos, nos países do Ocidente. A DMI exsudativa é uma das formas mais graves de apresentação da doença. Objectivos: Tendo em conta que esta doença parece partilhar alguns factores de risco com a síndrome metabólica, os objectivos neste trabalho foram procurar a existência de associações entre a síndrome metabólica e DMI exsudativa, avaliando também a composição corporal por antropometria e outros parâmetros bioquímicos. Métodos: Realizou-se um estudo caso controlo incluindo 81 indivíduos com mais de 50 anos, sendo 21 controlos e 60 casos com DMI exudativa em tratamento com Ranibizumab. A estes indivíduos foi aplicado um questionário estruturado (idade, estado civil, escolaridade, profissão e fumador actual ou anterior); posteriormente foram realizadas medições antropométricas (peso, altura, perímetro da cintura, perímetro da anca, bio-impedancia eléctrica), análises bioquímicas (colesterol total, colesterol LDL, apolipoproteína a1, apolipoproteína b, lipoproteina (a) e proteína C-reactiva) e exame oftalmológico para avaliação da DMI exsudativa. Os critérios usados para a classificação de síndrome metabólica foram os recomendados pela da Internacional Diabetes Federation (2005). Para a análise estatística foram calculadas as frequências absolutas e relativas para as variáveis categóricas, médias e desvios padrão para as variáveis contínuas. De seguida, foram efectuados testes t de student e testes de qui-quadrado. Por fim, foram feitas regressões logísticas pelo método passo-a-passo retrospectivo, usando como critério a razão de verosimilhança (backward stepwise - likelihood ratio). Considerou-se um nível de significância de 0,05.(...) / Dissertation abstract:Introduction: Age-related macular degeneration (AMD) is the leading cause of severe visual loss over the age of 50 in the western world. Exudative AMD is one of the most serious manifestations of the disease. Objectives: Having into account that this disease seems to share some of the risk factors with the metabolic syndrome, the main goal of this thesis was to look for the existence of associations between metabolic syndrome and Exudative AMD, as well as, associations between body composition, anthropometry and other biochemical parameters. Methods: A case control study was conducted and included 81 subjects with over 50 years, 21 controls and 60 cases with Exudative AMD that were treated with Ranibizumab. Subjects were submitted to a structured questionnaire (age, marital status, education, occupation and current or former smoker), anthropometric measurement (weight, height, waist circumference, hip circumference, bio-electrical impedance), biochemical analysis and an ophthalmologic evaluation. For the standard classification of metabolic syndrome the International Diabetes Federation (2005) criteria were used. For statistical analysis the absolute and relative frequencies for categorical variables, means and standard deviations for continuous variables were calculated. t Student test and chi-square were used. Finally, logistic regressions by the method step-by-step retrospective, using as criteria the likelihood ratio (backward stepwise - likelihood ratio) were performed. a significance level of 0.05 was considered. (...)