Characteristics Of Age-related Macular Degeneration Research Articles

LncRNA NORAD defects deteriorate the formation of age-related macular degeneration.

Long noncoding RNAs (lncRNAs) play important roles in the development of age-related macular degeneration (AMD). However, the effect of long non-coding RNA activated by DNA damage (NORAD) on AMD remains unknown. This study aimed to investigate the effect of NORAD on RPE cell senescence and degeneration. Irradiated adult retinal pigment epithelial cell line-19 (ARPE-19) and sodium iodate-treated mice were used as in vitro and in vivo AMD models. Results showed that irradiation-induced AMD characteristics of ARPE-19 and NORAD-knockdown aggravated cell cycle arrest in the G2/M phase, cell apoptosis and cell senescence along with the increased expression of phosphorylated P53 (p-P53) and P21. AMD factors C3, ICAM-1, APP, APOE, and VEGF-A were also increased by NORAD-knockdown. Moreover, NORAD-knockdown increased irradiation-induced reduction of mitochondrial homeostasis factors, (i.e., TFAM and POLG) and mitochondrial respiratory chain complex genes (i.e., ND1 and ND5) along with mitochondrial reactive oxygen species (ROS). We also identified a strong interaction of NORAD and PGC-1α and sirtuin 1 (SIRT1) in ARPE-19; that is, NORAD knockdown increases the acetylation of PGC-1α. In NORAD knockout mice, NORAD-knockout accelerated the sodium iodate-reduced retinal thickness reduction, function impairment and loss of retinal pigment in the fundus. Therefore, NORAD-knockdown accelerates retinal cell senescence, apoptosis, and AMD markers via PGC-1α acetylation, mitochondrial ROS, and the p-P53-P21signaling pathway, in which NORAD-mediated effect on PGC-1α acetylation might occur through the direct interaction with PGC-1α and SIRT1.

Food colloid-based delivery systems for tackling age-related macular degeneration by enhancing carotenoid bioavailability: A review

Over the past century, there has been a large increase in the life expectancy of people around the globe, which means there has been a rise in diseases of the elderly. Age-related macular degeneration (AMD) is a sight-threatening condition of the eye characterized by a loss of central vision. Research suggests that macular pigment carotenoids, such as lutein and zeaxanthin, may inhibit the onset of this disease by protecting the eye from damaging light and oxidation. These carotenoids are chemically reactive hydrophobic molecules with a low water-solubility, chemical stability, and bioavailability. Consequently, many carotenoid-rich foods and supplements are not absorbed by the human body and do not exhibit their beneficial effects on eye health. In this article, we review the prevalence and characteristics of AMD, the sources of macular pigment carotenoids in foods, the factors limiting their bioavailability, and analytical approaches available to study their bioavailability and bioactivity. We then discuss different strategies for increasing the concentration of bioavailable macular pigment carotenoids in the human diet, including dietary sources, supplements, functional foods, and excipient foods, with an emphasis on colloidal systems that can be used for this purpose.

Exploring association between pseudoexfoliation syndrome and ocular aging

PurposeWithin a population-based follow-up study, to examine the 10-year incidence of pseudoexfoliation syndrome (PEX), possible risk factors for PEX and its association with ocular aging of the cornea, lens and retina.MethodsThe baseline examination was conducted in 2006 on a random sample of 1,033 adult participants from Kaunas city (Lithuania) population of whom 631 had ophthalmic examination data at attendance of the 10-year follow-up in 2016. Detailed examination of the anterior and posterior segment of the eye was carried out. After diagnostic mydriasis PEX was diagnosed by the presence of typical grayish-white exfoliation material on the anterior capsule surface of the lens. The participants were divided to PEX and non-PEX groups.ResultsPEX prevalence increased from 9.8 to 34.2% from baseline to 10-year follow-up. Nuclear cataract was common both in the PEX group (66.7%) and in those without PEX (72.2%), but this difference did not reach statistically significantly increased risk of developing cataract in those with PEX (OR 1.2; p = 0.61). Central corneal thickness (CCT) was thinner in the PEX group (529 ± 34 μm) and in the oldest group (525 ± 36 μm) (p < 0.001). Compared to baseline, corneal curvature (CC) became flatter in both groups (7.6 ± 0.27 vs 7.7 ± 0.26 mm; p < 0.001) during the follow-up, but the difference did not reach significance between groups. Corneal astigmatism was most commonly with-the-rule in both groups (37 (50.0%) vs 148 (68.5%); p > 0.05). Age, sex and PEX had no influence on age-related macular degeneration distribution.ConclusionThe prevalence of PEX increased significantly with age in our population, with those with PEX having thinner and flatter corneae, but no difference in cataract and age-related macular degeneration characteristics.

Customized strategies for high-yield purification of retinal pigment epithelial cells differentiated from different stem cell sources

Retinal pigment epithelial (RPE) cell dysfunction and death are characteristics of age-related macular degeneration. A promising therapeutic option is RPE cell transplantation. Development of clinical grade stem-cell derived RPE requires efficient in vitro differentiation and purification methods. Enzymatic purification of RPE relies on the relative adherence of RPE and non-RPE cells to the culture plate. However, morphology and adherence of non-RPE cells differ for different stem cell sources. In cases whereby the non-RPE adhered as strongly as RPE cells to the culture plate, enzymatic method of purification is unsuitable. Thus, we hypothesized the need to customize purification strategies for RPE derived from different stem cell sources. We systematically compared five different RPE purification methods, including manual, enzymatic, flow cytometry-based sorting or combinations thereof for parameters including cell throughput, yield, purity and functionality. Flow cytometry-based approach was suitable for RPE isolation from heterogeneous cultures with highly adherent non-RPE cells, albeit with lower yield. Although all five purification methods generated pure and functional RPE, there were significant differences in yield and processing times. Based on the high purity of the resulting RPE and relatively short processing time, we conclude that a combination of enzymatic and manual purification is ideal for clinical applications.

Histologic Cell Shape Descriptors for the Retinal Pigment Epithelium in Age-Related Macular Degeneration: A Comparison to Unaffected Eyes.

PurposePhenotype alterations of the retinal pigment epithelium (RPE) are a main characteristic of age-related macular degeneration (AMD). Individual RPE cell shape descriptors may help to delineate healthy from AMD-affected cells in early disease stages.MethodsTwenty-two human RPE flatmounts (7 eyes with AMD [early, 3; geographic atrophy, 1; neovascular, 3); 15 unaffected eyes [8 aged ≤51 years; 7 aged >80 years)] were imaged at the fovea, perifovea, and near periphery (predefined sample locations) using a laser-scanning confocal fluorescence microscope. RPE cell boundaries were manually marked with computer assistance. For each cell, 11 shape descriptors were calculated and correlated with donor age, cell autofluorescence (AF) intensity, and retinal location. Statistical analysis was performed using an ensemble classifier based on logistic regression.ResultsIn AMD, RPE was altered at all locations (most pronounced at the fovea), with area, solidity, and form factor being the most discriminatory descriptors. In the unaffected macula, aging had no significant effect on cell shape factors; however, with increasing distance to the fovea, area, solidity, and convexity increased while form factor decreased. Reduced AF in AMD was significantly associated with decreased roundness and solidity.ConclusionsAMD results in an altered RPE with enlarged and deformed cells that could precede clinically visible lesions and thus serve as early biomarkers for AMD onset. Our data may also help guide the interpretation of RPE morphology in in vivo studies utilizing high-resolution single-cell imaging.Translational RelevanceOur histologic RPE cell shape data have the ability to identify robust biomarkers for the early detection of AMD-affected cells, which also could serve as a basis for automated segmentation of RPE sheets.

Age-related macular degeneration eyes presenting with cuticular drusen and reticular pseudodrusen

This study aimed to describe the clinical characteristics of age-related macular degeneration (AMD) eyes with both cuticular drusen (CD) and reticular pseudodrusen (RPD). Clinical records of patients diagnosed with CD or RPD with multimodal imaging was reviewed for patients diagnosed with both CD and RPD. The distribution patterns of CD (macular and diffuse type) and RPD (localized, intermediate, and diffuse type), presence of soft drusen, large drusen (> 200 µm), variant subretinal drusenoid deposits, and macular complications were investigated. Of the 220 eyes of 110 patients diagnosed with CD and 926 eyes of 463 patients diagnosed with RPD, 13 eyes of seven patients met the diagnostic criteria for both CD and RPD. The mean age at initial presentation was 71.4 ± 8.8 years and six patients were female. The mean subfoveal choroidal thickness was 143.8 ± 25.1 µm. The distribution of CD was of the macular type in all eyes. Distribution of RPD was localized in 11 eyes (84.6%) and intermediate in two eyes (15.4%). Soft drusen, large drusen, and variant subretinal drusenoid deposits were present in 13 (100%), 12 (92.3%) and, seven (53.8%) eyes, respectively. Macular neovascularization was observed in two eyes (15.4%). CD and RPD can coexist in eyes with AMD. Multimodal imaging should be used for AMD eyes with features suggestive of CD and RPD, considering the high likelihood of developing late AMD.

Profil épidémiologique et clinique de la dégénérescence maculaire liée à l’âge en milieu camerounais

To determine the epidemiological and clinical characteristics of age-related macular degeneration in patients in Yaoundé. A multicenter, analytical and cross-sectional study was carried out from March 2017 to June 2018 in four hospitals in the city of Yaoundé. Included were all consenting patients aged at least 40 years with an established diagnosis of AMD both clinically and by imaging. The data were analyzed with CSPro software version 7.0. Chi2 and Student's "t" tests in univariate mode and logistic regression in multivariate mode were used with a significance of P≤5%. Of the 9,989 patients who were seen during the 16-month study period, 38 met our inclusion criteria - a frequency of 0.4%. The sex ratio was 0.3, and the mean age was 68±11 years. As a function of geographic origin, AMD is more common in patients from the forest and mountain areas. The systemic comorbidities associated recorded were hypertension (47.4%) and diabetes (21.1%). After optical correction, 60 of 76 eyes (78.9%) had useful visual acuity according to WHO criteria, and the rate of blindness was 9.2%. Fundus examination revealed fewer lesions than optical coherence tomography, which demonstrated miliary drusen in 57 (75%) eyes, serous drusen in 27 (35.5%), retinal atrophy in 19 (25%) and neovascularization in 3 (3.9%). The clinical forms were dominated by age-related maculopathy, found in 45 eyes (69.2%), followed by atrophic AMD in 17 (26.2%) and finally by exudative AMD in 3 (4.6%), for a total of 65 out of 76 eyes. Age was related to the risk of MLA and atrophic AMD (P≤0.05). AMD is an uncommon pathology in our setting, predominant in women over the age of 60 years.

Inhibition of prolyl oligopeptidase: A promising pathway to prevent the progression of age-related macular degeneration

Dry age-related macular degeneration (AMD) is a currently untreatable vision threatening disease. Impaired proteasomal clearance and autophagy in the retinal pigment epithelium (RPE) and subsequent photoreceptor damage are connected with dry AMD, but detailed pathophysiology is still unclear. In this paper, we discover inhibition of cytosolic protease, prolyl oligopeptidase (PREP), as a potential pathway to treat dry AMD. We showed that PREP inhibitor exposure induced autophagy in the RPE cells, shown by increased LC3-II levels and decreased p62 levels. PREP inhibitor treatment increased total levels of autophagic vacuoles in the RPE cells. Global proteomics was used to examine the phenotype of a commonly used cell model displaying AMD characteristics, oxidative stress and altered protein metabolism, in vitro. These RPE cells displayed induced protein aggregation and clear alterations in macromolecule metabolism, confirming the relevance of the cell model. Differences in intracellular target engagement of PREP inhibitors were observed with cellular thermal shift assay (CETSA). These differences were explained by intracellular drug exposure (the unbound cellular partition coefficient, Kpuu). Importantly, our data is in line with previous observations regarding the discrepancy between PREP’s cleaving activity and outcomes in autophagy. This highlights the need to further explore PREP’s role in autophagy so that more effective compounds can be designed to battle diseases in which autophagy induction is needed. The present work is the first report investigating the PREP pathway in the RPE and we predict that the PREP inhibitors can be further optimized for treatment of dry AMD.

Sleeping pattern and activities of daily living modulate protein expression in AMD.

Degeneration of macular photoreceptors is a prominent characteristic of age-related macular degeneration (AMD) which leads to devastating and irreversible vision loss in the elderly population. In this exploratory study, the contribution of environmental factors on the progression of AMD pathology by probing the expression of candidate proteins was analyzed. Four hundred and sixty four participants were recruited in the study comprising of AMD (n = 277) and controls (n = 187). Genetics related data was analyzed to demonstrate the activities of daily living (ADL) by using regression analysis and statistical modeling, including contrast estimate, multinomial regression analysis in AMD progression. Regression analysis revealed contribution of smoking, alcohol, and sleeping hours on AMD by altered expression of IER-3, HTRA1, B3GALTL, LIPC and TIMP3 as compared to normal levels. Contrast estimate supports the gender polarization phenomenon in AMD by significant decreased expression of SLC16A8 and LIPC in control population which was found to be unaltered in AMD patients. The smoking, food habits and duration of night sleeping hours also contributed in AMD progression as evident from multinomial regression analysis. Predicted model (prediction estimate = 86.7%) also indicated the crucial role of night sleeping hours along with the decreased expression of TIMP-3, IER3 and SLC16A8. Results revealed an unambiguous role of environmental factors in AMD progression mediated by various regulatory proteins which might result in intermittent AMD phenotypes and possibly influence the outcome of anti-VEGF treatment.

Hypothetical pathogenesis of age-related macular degeneration and pachychoroid diseases derived from their genetic characteristics.

Genetic studies have investigated the pathogenesis of age-related macular degeneration (AMD). The pachychoroid concept has recently garnered attention as a possible explanation for AMD pathogenesis; the genetic characteristics of pachychoroid diseases have also been elucidated. In this review, we summarize previously reported genetic characteristics of AMD and pachychoroid diseases, and analyze these data to understand the pathogenesis of AMD and pachychoroid diseases. Previous studies show that VIPR2 and the CFH I62V A allele promote development of pachychoroid and central serous chorioretinopathy (CSC), while the CFH I62V G allele promotes development of drusen, pachychoroid neovasculopathy (PCN/PNV), and AMD. ARMS2/HTRA1 also promotes development of drusen, PCN/PNV, and AMD. TNFRSF10A and GATA5 are associated with CSC but not with pachychoroid, and TNFRSF10A is associated with AMD that includes PCN/PNV. These genetic characteristics suggest the following mechanisms of developing AMD and pachychoroid diseases. VIPR2 and the CFH I62V A allele promote pachychoroid development, which can result in CSC development. The CFH I62V G allele promotes a common step during PCN/PNV and AMD development induced by pachychoroid or drusen, such as damage of Bruch's membrane or retinal pigment epithelium (RPE). ARMS2/HTRA1 also promotes damage of Bruch's membrane or RPE, while the association with drusen formation is stronger in ARMS2/HTRA1 than in CFH. TNFRSF10A and GATA5 promote blood-retinal-barrier breakdown to induce CSC, which could lead to PCN/PNV development. Furthermore, recently reported genetic associations with the natural course of CSC suggest the importance of reconsidering the subtype classification of CSC. These associations would enable the development of personalized/precision medicine for CSC and.

An in vitro model of chronic wounding and its implication for age-related macular degeneration.

Degeneration of the retinal pigment epithelium (RPE) plays a central role in age-related macular degeneration (AMD). Throughout life, RPE cells are challenged by a variety of cytotoxic stressors, some of which are cumulative with age and may ultimately contribute to drusen and lipofuscin accumulation. Stressors such as these continually damage RPE cells resulting in a state of chronic wounding. Current cell-based platforms that model a state of chronic RPE cell wounding are limited, and the RPE cellular response is not entirely understood. Here, we used the electric cell-substrate impedance sensing (ECIS) system to induce a state of acute or chronic wounding on differentiated human fetal RPE cells to analyze changes in the wound repair response. RPE cells surrounding the lesioned area employ both cell migration and proliferation to repair wounds but fail to reestablish their original cell morphology or density after repetitive wounding. Chronically wounded RPE cells develop phenotypic AMD characteristics such as loss of cuboidal morphology, enlarged size, and multinucleation. Transcriptomic analysis suggests a systemic misregulation of RPE cell functions in bystander cells, which are not directly adjacent to the wound. Genes associated with the major RPE cell functions (LRAT, MITF, RDH11) significantly downregulate after wounding, in addition to differential expression of genes associated with the cell cycle (CDK1, CDC6, CDC20), inflammation (IL-18, CCL2), and apoptosis (FAS). Interestingly, repetitive wounding resulted in prolonged misregulation of genes, including FAS, LRAT, and PEDF. The use of ECIS to induce wounding resulted in an over-representation of AMD-associated genes among those dysregulated genes, particularly genes associated with advanced AMD. This simple system provides a new model for further investigation of RPE cell wound response in AMD pathogenesis.

Mitochondrial oxidative stress in the retinal pigment epithelium (RPE) led to metabolic dysfunction in both the RPE and retinal photoreceptors

Age-related macular degeneration (AMD) is the leading cause of vision loss in the western world. Recent evidence suggests that RPE and photoreceptors have an interconnected metabolism and that mitochondrial damage in RPE is a trigger for degeneration in both RPE and photoreceptors in AMD. To test this hypothesis, this study was designed to induce mitochondrial damage in RPE in mice to determine whether this is sufficient to cause RPE and photoreceptor damage characteristic of AMD. In this study, we conditionally deleted the gene encoding the mitochondrial antioxidant enzyme, manganese superoxide dismutase (MnSOD encoded by Sod2) in the retinal pigment epithelium (RPE) of albino BALB/cJ mice. VMD2-Cre;Sod2flox/flox BALB/cJ mice were housed in either 12-h dark, 12-h 200 lux white lighting (normal light), or 12-h dark, 12-h <10 lux red lighting (dim light). Electroretinography (ERG) and spectral-domain optical coherence tomography (SD-OCT) were performed to assess retinal function and morphology. Immunofluorescence was used to examine protein expression; quantitative RT-PCR was used to measure gene expression. Sod2 knockout (KO) mice had reduced RPE function with age and increased oxidative stress compared to wild type (WT) controls as expected by the cell-specific deletion of Sod2. This was associated with alterations in RPE morphology and the structure and function of RPE mitochondria. In addition, data show a compensatory increase in RPE glycolytic metabolism. The metabolic shift in RPE correlated with severe disruption of photoreceptor mitochondria including a reduction in TOMM20 expression, mitochondrial fragmentation, and reduced COXIII/β-actin levels. These findings demonstrate that mitochondrial oxidative stress can lead to RPE dysfunction and metabolic reprogramming of RPE. Secondary to these changes, photoreceptors also undergo metabolic stress with increased mitochondrial damage. These data are consistent with the hypothesis of a linked metabolism between RPE and photoreceptors and suggest a mechanism of retinal degeneration in dry AMD.

Morphologic features of large choroidal vessel layer: age-related macular degeneration, polypoidal choroidal vasculopathy, and central serous chorioretinopathy.

To compare choroidal vascular characteristics of age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV), and central serous chorioretinopathy (CSC) by qualitative and quantitative analyses using swept-source en face optical coherence tomographic (OCT) images. Eyes with non-neovascular AMD (n = 32), neovascular AMD (n = 30), thick and thin choroid PCV (n = 33 and 27), and CSC (n = 34) were enrolled. Subfoveal choroidal thickness (SFCT) and the presence and patterns of pachyvessels were assessed. En face images of the large choroidal vessel layer were converted to binary images for the analysis of vascular density. Pachyvessels were identified in 8 (25%), 14 (46%), 28 (85%), 26 (96%), and 34 (100%) non-neovascular AMD, neovascular AMD, thin choroid PCV, thick choroid PCV, and CSC eyes, respectively (P < 0.001). The pattern of pachyvessels was focal in non-neovascular AMD (100%), neovascular AMD (79%), and thin choroid PCV (89%) while the pattern was mostly diffuse in CSC (88%) and thick choroid PCV (81%). The mean choroidal vascular density in a 6 × 6mm2 macular area of each group was 45.3%, 46.9%, 47.0%, 52.5%, and 54.8%, respectively (P < 0.001). Post hoc analysis revealed significantly higher vascular density in CSC compared with other types (all P < 0.001) except PCV with thick choroid (P = 0.066). Similarities in vascular density of the large choroidal vessel layer and pachyvessel pattern were between CSC and thick choroid PCV and between AMD and thin choroid PCV, suggesting common pathophysiology involving choroidal changes in these eyes.

The role of complement system and other inflammatory factors in the development of age-related macular degeneration

The article is a review of literature on the role of complement system and inflammatory factors in the development of age-related macular degeneration. The review uses materials of domestic and foreign researchers. The clinical characteristics of age-related macular degeneration are presented, the role of genetic factors, complement factors, biomarkers of inflammation and alternative pathway of complement activation in the pathogenesis and risk of age-related macular degeneration is determined. Age-related macular degeneration is a chronic progressive multifactorial disease that affects macular area of the retina and is the main cause of loss of central vision in patients of older age group. The most important genetic factors are chromosome 1 (1q32) including complement factor H and complement factor H related genes and chromosome 10 (10q31). Variants associated with a moderate effect on developmental risk were identified in C3, complement factor I and complement factor B genes. In the pathogenesis of age-related macular degeneration, the key role is played by the damaged regulation of the alternative complement pathway. Single nucleotide polymorphisms in complement genes that affect the risk of development of age-related macular degeneration are predominantly involved in the alternative pathway of activation of the complement system. In pathomorphological studies, the initial localization of the pathological process of this pathology was established to be a complex of retinal pigment epithelium, Bruch’s membrane, and choriocapillaries followed by loss of photoreceptor function. The review of studies of systemic inflammatory biomarkers, cytokines, vascular endothelial growth factors in peripheral blood, blood serum, aqueous humour at various stages and forms of age-related macular degeneration is presented.

ID2 protects retinal pigment epithelium cells from oxidative damage through p-ERK1/2/ID2/NRF2

Age-related macular degeneration (AMD) is the leading cause of blindness during aging. The degeneration of retinal pigment epithelium (RPE) is the main pathologic characteristic of AMD. ID2 is a member of the Inhibitor of DNA binding proteins (ID) family and is involved in regulation of cell proliferation and differentiation. However, currently the role of ID2 in oxidative injury response in RPE cells remains unknown. Here we showed that oxidative stress increased ID2 expression in RPE cells. Knockdown of ID2 promoted cell apoptosis and increased ROS level in RPE cells that were subjected to oxidative damage. In addition, over-expression of ID2 attenuated the oxidative damage response in RPE cells. Mechanistically, ID2 protected RPE cells from oxidative damage through activating NRF2. Furthermore, phosphorylation of ERK1/2 positively regulated the protective function of ID2. Finally, we confirmed that the oxidative damage increased Id2 expression and over-expression of Id2 elevated Nrf2 expression in primary mouse RPE cells. Therefore, ID2 protects RPE cells from oxidative damage through the p-ERK1/2/ID2/NRF2 pathway. Our study contributes to a better understanding of the mechanisms underlying oxidative stress in AMD and may present a new strategy for AMD treatment.

Choroidal Angioarchitecture According to Ultra-Widefield Indocyanine Green Angiography in Age-Related Macular Degeneration

Purpose: To examine choroidal blood flow using ultra-widefield indocyanine green angiography (ICGA) in patients with age-related macular degeneration (AMD) and to compare these characteristics with those in normal controls. Methods: We reviewed the findings of ultra-widefield ICGA in patients with AMD and age-matched normal controls for comparison between April 2016 and November 2016. The primary outcome measure was the comparison of the vascular patterns of the choroid in the venous phase. The secondary outcome was abnormalities of the arterioles or venules in the macula in each group. Results: The study included 113 eyes of 84 patients with AMD (26 eyes with nonneovascular AMD, 58 eyes with typical AMD, and 29 eyes with polypoidal choroidal vasculopathy), 50 fellow eyes without the clinical characteristics of AMD, and 27 eyes of 24 age-matched normal controls. Patients with AMD were more likely than normal controls to have the region of greater hyperfluorescence of the ICGA with asymmetry of choroidal vasculature or abnormality of the vessels in the macula. Abnormalities of the vessels in the macula could be classified into 3 types: a small vortex vein, anastomosis between the choroidal veins, and the large choroidal vessels. Conclusions: Using ultra-widefield ICGA, we found that having an asymmetry of choroidal venules or abnormality of the vessels in the macula may be a predictive risk factor for the development or progression of AMD. These findings also support the association of alterations in choroidal hemodynamics with the pathogenesis of AMD.

Interactome Mapping Guided by Tissue-Specific Phosphorylation in Age-Related Macular Degeneration.

The current study aims to determine the molecular mechanisms of age-related macular degeneration (AMD) using the phosphorylation network. Specifically, we examined novel biomarkers for oxidative stress by protein interaction mapping using in vitro and in vivo models that mimic the complex and progressive characteristics of AMD. We hypothesized that the early apoptotic reactions could be initiated by protein phosphorylation in region-dependent (peripheral retina vs. macular) and tissue-dependent (retinal pigment epithelium vs. retina) manner under chronic oxidative stress. The analysis of protein interactome and oxidative biomarkers showed the presence of tissue- and region-specific post-translational mechanisms that contribute to AMD progression and suggested new therapeutic targets that include ubiquitin, erythropoietin, vitronectin, MMP2, crystalline, nitric oxide, and prohibitin. Phosphorylation of specific target proteins in RPE cells is a central regulatory mechanism as a survival tool under chronic oxidative imbalance. The current interactome map demonstrates a positive correlation between oxidative stress-mediated phosphorylation and AMD progression and provides a basis for understanding oxidative stress-induced cytoskeletal changes and the mechanism of aggregate formation induced by protein phosphorylation. This information could provide an effective therapeutic approach to treat age-related neurodegeneration.

All-trans-retinal dimer formation alleviates the cytotoxicity of all-trans-retinal in human retinal pigment epithelial cells.

Effective clearance of all-trans-retinal (atRAL) from retinal pigment epithelial (RPE) cells is important for avoiding its cytotoxicity. However, the metabolism of atRAL in RPE cells is poorly clarified. The present study was designed to analyze metabolic products of atRAL and to compare the cytotoxicity of atRAL versus its derivative all-trans-retinal dimer (atRAL-dimer) in human RPE cells. We found that all-trans-retinol (atROL) and a mixture of atRAL condensation metabolites including atRAL-dimer and A2E were generated after incubating RPE cells with atRAL for 6h, and the amount of atRAL-dimer was significantly higher than that of A2E. In the eyes of Rdh8-/- Abca4-/- mice, a mouse model with defects in retinoid cycle that displays some symbolic characteristics of age-related macular degeneration (AMD), the level of atRAL-dimer was increased compared to wild-type mice, and was even much greater than that of A2E & isomers. The cytotoxicity of atRAL-dimer was reduced compared with its precursor atRAL. The latter could provoke intracellular reactive oxygen species (ROS) overproduction, increase the mRNA expression of several oxidative stress related genes (Nrf2, HO-1, and γ-GCSh), and induce ΔΨm loss in RPE cells. By contrast, the abilities of atRAL-dimer to induce intracellular ROS and oxidative stress were much weaker versus that of concentration-matched atRAL, and atRAL-dimer exhibited no toxic effect on mitochondrial function at higher concentrations. In conclusion, the formation of atRAL-dimer during atRAL metabolic process ameliorates the cytotoxicity of atRAL by reducing oxidative stress.

Automated detection and segmentation of drusen in retinal fundus images

The druse, an abnormal yellow/white deposit on retina, is a dominant characteristic of age-related macular degeneration (AMD) which is a retinal disorder associated with age. The early detection of drusen is useful for ophthalmologists to diagnose the patients that suffer from AMD. An automated method has been proposed in this work to detect and segment drusen using retinal fundus images by (i) gradient based segmentation to find true edges of drusen, (ii) connected component labeling to remove suspicious pixels from drusen region and (iii) edge linking to connect all labeled pixels into a meaningful boundary. The proposed method outperforms other existing methods in detection of drusen with an accuracy/sensitivity/specificity of 96.17/89.81/99.00 on two publicly available retinal image databases. In order to grade the severity of AMD, the detected drusen by the proposed method are further quantified into small, intermediate and large with an accuracy of 88.46, 98.55, and 88.37%, respectively.

Quality of Life in Patients Suffering from Active Exudative Age-Related Macular Degeneration: The EQUADE Study

Purpose: Age-related macular degeneration (AMD) is the main cause of visual loss in the elderly population. With the use of anti-vascular endothelial growth factor, the visual outcomes of exudative AMD patients have been improved. This study was aimed at assessing the quality of life (QoL) of exudative AMD patients treated with ranibizumab and at determining its drivers in a real-life setting. Methods: We performed a national, cross-sectional, observational survey based on questionnaires sent to members of French associations relative to AMD between December 2012 and March 2013. Patients suffering from exudative AMD with at least one intravitreal injection of ranibizumab within the last 6 months were included. Demographics, AMD characteristics, visual acuity (VA) and past and ongoing treatments were collected. The 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) was self-administered. A multivariate model was used to identify QoL drivers. Results: 416 questionnaires fulfilled the complete criteria for both QoL and cost analyses. The mean age of exudative AMD patients was 78.0 years and bilateral involvement was reported in 60.4%. The overall mean QoL score was 53.4. Mental health, driving and role difficulties were the most widely affected domains. After bivariate analyses, long-term illness status, worse VA and higher number of unpaid aids were associated with worse QoL, with odds ratios of 2.4, 5.2 and 11.6, respectively. The mean cost per year and per patient was 1,741 EUR. The main components of costs were aids and services and the purchase of visual equipment. Conclusions: The main predictors of QoL in exudative AMD patients treated with ranibizumab are VA outcomes, home healthcare and social services provided to the patients.