Age-related Macular Degeneration Eyes Research Articles

Lutein: A Valuable Ingredient of Fruit and Vegetables

Lutein is a human serum carotenoid which is not synthesized by humans and thus must be obtained by the ingestion of food containing it such as fruits and vegetables. Lutein is present in different forms in those foods as all-trans-lutein, cis-lutein, epoxi-lutein, and lutein linked to proteins. It discusses if the intake of lutein or diets supplemented with lutein or diets rich in fruits and vegetables are important in the prevention of diseases like some cancers, cardiovascular diseases, etc., that may be affected by the antioxidant effect of lutein; or in the prevention of age-related macular degeneration and other eye diseases. The concentration of lutein in fruits and vegetables depends on the species. We've included the concentration of lutein in 74 species reported by different authors since 1990. Currently the quantification of lutein is mainly performed by HPLC, but more investigations into a quantification method for lutein, lutein isomers, and epoxi-lutein are necessary. Improvement of lutein extraction methods is important as well. Methods commonly used in the vegetable and fruit industry like heat treatment, storage conditions, etc. can change lutein concentrations; other factors depend on the plant, for instance the variety, the stage of maturity, etc.

Detection of CX3CR1 single nucleotide polymorphism and expression on archived eyes with age-related macular degeneration.

There is a significant genetic component in age-related macular degeneration (AMD). CX3CR1, which encodes the fractalkine (chemokine, CX3CL1) receptor, has two single nucleotide polymorphisms (SNPs): V249I and T280M. These SNPs are correlated with other aged-related diseases such as atherosclerosis. We have reported an association of CX3CR1 SNP and AMD. In this study we examined CX3CR1 SNP frequencies and protein expression on archived sections of AMD and normal eyes. We microdissected non-retinal, peripheral retinal and macular cells from archived slides of eyes of AMD patients and normal subjects. CX3CR1 SNP typing was conducted by PCR and restriction fragment length polymorphism analysis. CX3CR1 transcripts from retinal cells were also measured using RT-PCR. CX3CR1 protein expression was evaluated using avidin-biotin complex immunohistochemistry. We successfully extracted DNA from 32/40 AMD cases and 2/2 normal eyes. Among the 32 AMD cases, 18 had neovascular AMD and 14 had non-neovascular AMD. The M280 allele was detected in 19/64 (32 cases x2) with a frequency of 29.7%, which was significantly higher as compared to the frequency in the normal population (11.2%). We detected CX3CR1 expression in the various retinal cells. CX3CR1 transcript and protein levels were diminished in the macular lesions. This study successfully analyzed CX3CR1 SNP and transcript expression in microdissected cells from archived paraffin fixed slides. Our data suggest that the M280 allele, a SNP resulting in aberrant CX3CR1 and CX3CL1 interaction, as well as lowered expression of macular CX3CR1, may contribute to the development of AMD.

DECREASED ARTERIAL DYE-FILLING AND VENOUS DILATION IN THE MACULAR CHOROID ASSOCIATED WITH AGE-RELATED MACULAR DEGENERATION

To compare the angioarchitecture of choroidal arteries and veins in patients with age-related macular degeneration (AMD) to the angioarchitecture of age-matched normal subjects using indocyanine green (ICG) angiography. ICG angiography was performed in 35 consecutive AMD patients and 18 normal age-matched volunteers with a fundus ICG camera. ICG video images, including the arterial and venous phases, were quantitatively analyzed using image analyzing software. In patients with AMD, the choroidal arterioles are dilated, fewer, run a straighter course, and possess fewer bifurcations. The number of choroidal arteries and the macular fluorescent intensity in the arterial phase of choroidal filling was significantly less in patients with AMD as compared to age-matched normal controls (P = 0.008). The mean and maximum caliber of choroidal veins in the macula was dilated in AMD eyes than in age-matched normal control eyes (P < 0.001). There was no statistically significant difference in arterial dye filling or venous caliber observed in AMD eyes, with or without choroidal neovascular membrane (CNV). Choroidal arterial perfusion in the macula was significant decreased in eyes with AMD with and without CNV, and was associated with choroidal venous dilation. These observations implicate poor choroidal perfusion of the macula in the pathogenesis of AMD.

Polypoidal choroidal vasculopathy and photodynamic therapy with verteporfin

We evaluated, in a nonrandomised, institutional, prospective study, the efficacy of photodynamic therapy (PDT) with verteporfin in age-related macular degeneration (AMD) eyes with polypoidal choroidal vasculopathy (PCV) and subfoveal exudation. A prospective clinical and angiographic study was done in 40 consecutive eyes with PCV treated with PDT using masked best-corrected visual acuity (VA) and fluorescein and indocyanine green angiographic features at baseline and over 2 years. Twenty-one eyes completed 1-year follow-up and showed, after a mean 2.9 PDT sessions, VA improvement in 12 eyes, no change in five eyes, and VA decrease in four eyes. Leakage was absent at the retinal and choroidal level in 14 eyes at 1 year. Recurrence occurred in one eye during the first year. Six eyes completed 2 years of follow-up and showed, after a mean 4 PDT sessions, VA improvement in five eyes and VA decrease in one eye. Leakage was absent at the retinal and choroidal level in five eyes. Recurrence occurred in four of these six eyes during the second year of follow-up. No serious adverse events were observed during the 2 years of follow-up. PDT with verteporfin was shown to be safe and effective for treating AMD eyes with PCV with subfoveal involvement. VA improvement and absence of leakage were achieved, respectively, in 57.1% and 66.6% of the eyes at 1 year. Recurrences were more frequent during the second year of follow-up.

Drusen classification in bilateral drusen and fellow eye of exudative age-related macular degeneration

To assess the value of the modified international classification system in screening high-risk patients with bilateral age-related maculopathy (ARM) from those with lower risk characteristics. In total, 164 digital images of 106 patients with either bilateral ARM (group A) or the fellow eyes of unilateral exudative age-related macular degeneration (AMD) (Group B) were included. Patients with no signs of ARM in both eyes or those with bilateral late AMD were excluded. The images were randomised and then graded by two masked ophthalmologists based on the modified International Classification of ARM. The interobserver consistency between the two graders was high with a Kappa value of 0.82 (SE 0.34, P<0.0001). There were no significant differences in the distribution of the stages of ARM between the two subgroups. Stage 3 was the most common stage in each group for both graders followed by stage 2a in the bilateral drusen group. Stages 1a, 2a and 2b were equally the next common stage in the fellow eye of chordial neovascularisation group. A screening system based on clinical characteristics would be of value in risk prediction in a clinical setting. Type of Drusen alone, as identified by the modified International grading system, may not be reliably predictive in screening for patients who are at high risk of developing choroidal neovascularisation.

Crystallin distribution in Bruch's membrane–choroid complex from AMD and age-matched donor eyes

Crystallins were consistently found in a recent proteomic analysis of drusen from age-related macular degeneration (AMD) donor eyes. Here we compare the distribution of several crystallins in drusen, Bruch's membrane and choroid from AMD and non-AMD age-matched control eyes. Immunohistochemistry and Western blots of tissue samples were performed using antibodies to αA- and αB-crystallins. Bruch's membrane, drusen and the subjacent choroidal connective tissue from AMD tissues showed greater immunoreactivity for αA- and αB-crystallins than were observed in normal age-matched control tissues. Western blots also demonstrated more intense αA- and αB-crystallin signals from AMD tissues than were present in age-matched controls. These data indicate that αA- and αB-crystallins accumulate in Bruch's membrane and choroidal connective tissues to a greater degree in AMD than in normal aging. These findings suggest that the accumulation of these small heat shock proteins at this critical interface below the RPE reflects a disease-related stress response manifested during the progression of AMD.

Photodynamic therapy for AMD: smaller is better!

Photodynamic therapy for AMD: smaller is better!

The involvement of sequence variation and expression of CX3CR1 in the pathogenesis of age-related macular degeneration.

This study examined the association between the sequence variation/expression of CX3CR1, a chemokine receptor, and age-related macular degeneration (AMD). Peripheral blood from 85 AMD patients and 105 subjects without AMD (controls), as well as ocular tissue from 40 pathological sections with AMD and two normal eye sections, were screened for V249I and T280M, two single nucleotide polymorphisms (SNPs) in CX3CR1. An increased prevalence, with the highest odds ratio of 3.57, of the I249 and M280 carriers was found among the AMD cases as compared with the controls. When comparing CX3CR1 expression in the archived eye sections, CX3CR1 transcripts were not detectable in the maculae of AMD eyes bearing T/M280; however, transcripts were detected in the maculae of normal eyes bearing T/T280 or T/M280 as well as in the AMD maculae bearing T/T280. Furthermore, lower CX3CR1 protein expression was observed in the maculae of AMD eyes bearing T/M280 compared with the controls bearing T/T280. The I249 and M280 alleles result in a lowered number of receptor binding sites and a decreased ligand affinity. Our data suggest that a decrease, caused by sequence variation and/or lower CX3CR1 expression, in CX3CR1-induced cellular activities could contribute to AMD development.

Stabilization of visual acuity with photodynamic therapy in eyes with chorioretinal anastomoses.

(1) To evaluate, in a non-randomized, institutional, prospective study, the efficacy of photodynamic therapy with Visudyne (PDT) in neovascular age-related macular degeneration (AMD) eyes with chorioretinal anastomoses (CRA). (2) To review, in a retrospective study and for comparison, the natural evolution of neovascular AMD eyes with CRA. Prospective clinical and angiographic study of 17 consecutive eyes with CRA, treated with PDT. Retrospective clinical and angiographic study of the natural course of 17 consecutive patients with CRA. Masked best-corrected visual acuity (VA) and angiographic features at baseline and during the period of one year were evaluated. The two groups presented similar characteristics at baseline regarding age, sex, initial VA, duration of follow-up and angiographic features. PDT-treated eyes showed, at 1-year follow-up, VA stabilization or improvement in 73.3% of the eyes, no cases with very severe VA loss, and no fluorescein leakage in 46.6% of the eyes. In contrast, at 1-year follow-up the natural evolution of CRA was characterized by severe or very severe VA loss in 69% of the eyes and statistically significant mean VA loss (P=0.001) with persistence of fluorescein leakage in all cases. The natural history of AMD eyes with CRA leads to progressive and dramatic VA loss, which is associated with blindness in most of the cases. PDT with verteporfin can offer some benefit to these patients, allowing VA stabilization or improvement in more than two thirds of the cases, at one year.

Mitochondria-derived reactive oxygen species mediate blue light-induced death of retinal pigment epithelial cells.

Throughout the lifetime of an individual, light is focused onto the retina. The resulting photooxidative stress can cause acute or chronic retinal damage. The pathogenesis of age-related macular degeneration (AMD), the leading cause of legal blindness in the developed world, involves oxidative stress and death of the retinal pigment epithelium (RPE) followed by death of the overlying photoreceptors. Evidence suggests that damage due to exposure to light plays a role in AMD and other age-related eye diseases. In this work a system for light-induced damage and death of the RPE, based on the human ARPE-19 cell line, was used. Induction of mitochondria-derived reactive oxygen species (ROS) is shown to play a critical role in the death of cells exposed to short-wavelength blue light (425 +/- 20 nm). ROS and cell death are blocked either by inhibiting the mitochondrial electron transport chain or by mitochondria-specific antioxidants. These results show that mitochondria are an important source of toxic oxygen radicals in blue light-exposed RPE cells and may indicate new approaches for treating AMD using mitochondria-targeted antioxidants.

Amyloid-β is found in drusen from some age-related macular degeneration retinas, but not in drusen from normal retinas

Immunohistochemical staining was performed with three Aβ antibodies on sections from nine normal eyes and nine eyes with age-related macular degeneration (AMD). The AMD eyes included three eyes with early AMD, three eyes with geographic atrophy, and three eyes with exudative disease. Five sections form each eye were evaluated. Aβ-positive deposits in drusen were identified using epifluorescence and confocal microscopy. Antibodies were pre-adsorbed with Aβ peptide to verify specificity. Some sections were also stained with PAS-hematoxylin to aid in identification of morphology. The Aβ antibody was optimized by detection of Aβ in amyloid plaques in Alzheimer’s disease. The Aβ label was blocked by pre-adsorption of antibody with Aβ peptide, verifying specificity. Four of the nine AMD specimens and none of nine controls contained Aβ-positive drusen. Two of the early AMD eyes had a few Aβ-positive drusen, each with a few Aβ-containing vesicles, and two of the geographic atrophy eyes had many Aβ-positive drusen with many Aβ-containing vesicles. In four of the nine eyes with AMD, Aβ was localized to a subset of drusen. Aβ in drusen correlated with the location of degenerating photoreceptors and retinal pigment epithelium.—Hans E. Grossniklaus

The Eger Macular Stressometer: pilot study

Purpose To evaluate the sensitivity of the Eger Macular Stressometer (EMS) for early screening of age-related macular degeneration (AMD) in a clinical practice. We examined the null hypothesis that AMD eyes have EMS recovery times (RTs) that do not differ from eyes with cataract, diabetic retinopathy, or glaucoma. Design The design of this study was a nonrandomized clinical trial. Methods Ninety-two eyes from 92 patients with vision 20/80 or better, age 50 and older, of either gender, and any ethnic origin, were recruited into one of four groups: AMD (30 eyes), normal or mild cataract (30 eyes), diabetic retinopathy (16 eyes), and glaucoma (16 eyes). Recovery times were obtained with the EMS, according to manufacturer’s instructions. Results The mean (SD) [median] RT for the AMD group was 11.8 (7.6) [9] seconds, the normal/cataract group 10.0 (4.3) [9] seconds, the diabetic retinopathy group 8.4 (3.0) [8] seconds, and glaucoma group 8.6 (2.4) [8] seconds. Recovery time did not appear to be related to group ( P = .58), age ( P = .50), visual acuity ( P = .52), or sex ( P = .23). Conclusion We found EMS RT distributions did not differ between AMD, cataract, diabetic retinopathy, and glaucoma groups. The EMS in its current form is not a sensitive screening tool for AMD. Further testing is needed to examine EMS sensitivity with other macular diseases such as central serous choroidopathy and diabetic macular edema.

Risk of age-related macular degeneration in eyes with macular drusen or hyperpigmentation: the Blue Mountains Eye Study cohort.

To quantify the 5-year risk of age-related macular degeneration (AMD) in eyes with different macular drusen characteristics (ie, size, type, location, and total area) or hyperpigmentation in a population-based cohort. The Blue Mountains Eye Study examined 3654 residents during 1992-1994; 2335 (75.1% of survivors) were reexamined during 1997-1999. Retinal photographs were graded using the Wisconsin Age-Related Maculopathy Grading System. Incident AMD lesions were defined by development of neovascular AMD or geographic atrophy in eyes without these lesions at baseline (eyes at risk). Age-adjusted relative risks (RRs) were determined. Generalized estimating equation models were used to estimate odds ratios, adjusting for the correlation between eyes and other AMD risk factors. Main Outcome Measure Incidence of AMD. Of the 4634 eyes at risk, 52 (1.1%) developed neovascular or atrophic AMD lesions over 5 years. In right eyes, presence vs absence of the following macular signs predicted AMD: drusen that were 125 micro m or larger (13.9 vs 0.6%; age-adjusted RR, 5.7; 95% confidence interval [CI], 3.6-9.0), indistinct soft or reticular drusen (23.2% vs 0.4%; RR, 9.9; 95% CI, 6.4-15.4), total drusen area of half the disc area or more (31.4% vs 0.6%; RR, 13.5; 95% CI, 8.0-22.8), and hyperpigmentation (14.4% vs 0.5%; RR, 8.0; 95% CI, 5.4-11.9). After adjusting for age, sex, and smoking status, eyes with these signs at baseline had a high likelihood of developing AMD. Eyes with Age-Related Eye Disease Study categories 3 and 4 were 5 times more likely to develop AMD compared with eyes in categories 1 and 2. This study quantifies the 5-year risk of AMD in eyes with macular drusen and hyperpigmentation.

The changes of multifocal electroretinography in the early stage of photodynamic therapy for choroidal neovascularization.

To investigate short-term changes in the multifocal electroretinography (ERG) recordings after photodynamic therapy (PDT) for choroidal neovascularization (CNV), 16 patients (17 eyes) with classic CNV confirmed by fluorescein angiography (FA) and indocyanine green angiography (ICGA), including 11 cases (12 eyes) of exudative age-related macular degeneration (AMD), two cases (two eyes) of pathological myopia and three cases (three eyes) of idiopathic causes, were treated using PDT with verteporfin. The multifocal ERGs of these patients were tested with VERIS Science 4.0 imaging system. The latencies and average response densities of all six ring retinal regions were measured and compared before PDT and 3 or 7 days after PDT. The latencies and amplitude densities of the N1 and P1 waves in all six rings remained unchanged at 3 or 7 days post-treatment (p > 0.05). Therefore, there is no significant evidence to suggest an adverse effect from PDT for classic CNV on the outer retinal function in the early stage of treatment, with the aid of the multifocal ERG recordings.

Photodynamic therapy for choroidal neovascular disease: photosensitizers and clinical trials

Photodynamic therapy with visudyne for choroidal neovascular disease (CNV) has proved effective at preventing moderate to severe visual loss in eyes with subfoveal predominantly classic CNV or occult-only CNV caused by age-related macular degeneration (AMD) and in eyes with subfoveal CNV caused by pathologic myopia. PDT is not meant to improve visual acuity. It should, therefore, be used in eyes with potentially useful macular vision. Clinical trials are under way to evaluate the usefulness of early retreatment with PDT and visudyne at 6 weeks for reducing the risk for moderate visual loss in eyes with predominantly classic CNV, the efficacy of PDT with visudyne for the treatment of AMD in eyes with minimally classic CNV, and the efficacy of PDT for use with SnET2, lutetium texaphyrin, and Npe6.

Resonance Raman measurement of macular carotenoids in normal subjects and in age-related macular degeneration patients

Purpose Dietary carotenoids lutein and zeaxanthin may play a protective role against visual loss from age-related macular degeneration (AMD) through antioxidant and light screening mechanisms. We used a novel noninvasive objective method to quantify lutein and zeaxanthin in the human macula using resonance Raman spectroscopy and compared macular pigment levels in AMD and normal subjects. Design Observational study of an ophthalmology clinic-based population. Participants and controls Ninety-three AMD eyes from 63 patients and 220 normal eyes from 138 subjects. Methods Macular carotenoid levels were quantified by illuminating the macula with a low-power argon laser spot and measuring Raman backscattered light using a spectrograph. This technique is sensitive, specific, and repeatable even in subjects with significant macular pathologic features. Main outcome measure Raman signal intensity at 1525 cm −1 generated by the carbon-carbon double-bond vibrations of lutein and zeaxanthin. Results Carotenoid Raman signal intensity declined with age in normal eyes ( P < 0.001). Average levels of lutein and zeaxanthin were 32% lower in AMD eyes versus normal elderly control eyes as long as the subjects were not consuming high-dose lutein supplements ( P = 0.001). Patients who had begun to consume supplements containing high doses of lutein (≥4 mg/day) regularly after their initial diagnosis of AMD had average macular pigment levels that were in the normal range ( P = 0.829) and that were significantly higher than in AMD patients not consuming these supplements ( P = 0.038). Conclusions These findings are consistent with the hypothesis that low levels of lutein and zeaxanthin in the human macula may represent a pathogenic risk factor for the development of AMD. Resonance Raman measurement of macular carotenoid pigments could play an important role in facilitating large-scale prospective clinical studies of lutein and zeaxanthin protection against AMD, and this technology may someday prove useful in the early detection of individuals at risk for visual loss from AMD.

Aberrant accumulation of EFEMP1 underlies drusen formation in Malattia Leventinese and age-related macular degeneration.

Malattia Leventinese (ML), an inherited macular degenerative disease, is closely reminiscent of age-related macular degeneration (AMD), the most common cause of incurable blindness. Both ML and AMD are characterized by extracellular deposits known as drusen between the retinal pigment epithelium (RPE) and Bruch's membrane. The mechanism underlying drusen formation is unknown. An Arg to Trp mutation in a gene of unknown function, EFEMP1, is responsible for ML, indicating EFEMP1 may be important in drusen formation. Here, we show that wild-type EFEMP1 is a secreted protein whereas mutant EFEMP1 is misfolded, secreted inefficiently, and retained within cells. In normal eyes, EFEMP1 is not present at the site of drusen formation. However, in ML eyes, EFEMP1 accumulates within the RPE cells and between the RPE and drusen, but does not appear to be a major component of drusen. Furthermore, in AMD eyes, EFEMP1 is found to accumulate beneath the RPE immediately overlaying drusen, but not in the region where there is no apparent retinal pathology observed. These data present evidence that misfolding and aberrant accumulation of EFEMP1 may cause drusen formation and cellular degeneration and play an important role in the etiology of both ML and AMD.

Surgical Removal of Submacular Choroidal Neovascularization: A Clinicopathologic Study and Factors Influencing Visual Outcomes

Clinical and histopathologic characteristics of submacular choroidal neovascular membranes (CNV) are investigated and features predictive of postoperative complications or poorer visual outcome following CNV removal are identified. Records of patients who underwent submacular CNV removal at Bascom Palmer Eye Institute between 1991 and 1998 are reviewed. At 6 months postoperatively, vision was stable or improved in 19/26 (73%) eyes, with 15/26 (58%) having 20/200 to 20/400 vision. Postoperative retinal detachment occurred more commonly in age-related macular degeneration eyes (AMD) (P <0.05), and CNV recurrence occurred most frequently in presumed ocular histoplasmosis (POHS) eyes (P <0.05). The one eye with an extrafoveal CNV in-growth site had the largest improvement in vision postoperatively. Choroid in the specimen was associated with worse postoperative vision (P = 0.05). Submacular CNV removal achieves visual stabilization at the 20/200 to 20/400 level in most eyes. Potential risk factors for postoperative complications or poorer visual outcome include the underlying disease process, subfoveal in-growth site of CNV, and presence of choroid in the specimen.

Hyperfluorescent Areas Observed in Late-phase Indocyanine Green Angiography (IA) of IA Findings in Fellow Eyes of Unilateral Age-related Macular Degeneration

Purpose: To evaluate abnormal findings of indocyanine green angiography (IA), predisposing to exudative age-related macular degeneration (AMD) in fellow eyes of unilateral exudative AMD. Methods: We categorized 83 fellow eyes of unilateral exudative AMD into two groups, ie, 55 eyes with early age-related maculopathy, and 28 eyes with normal aging. We performed IA. Results: The late phase of IA demonstrated hyperfluorescent areas in 27 of the 83 fellow eyes, which included 22 of the 55 eyes with early age-related maculopathy and five eyes of 28 eyes with normal aging. The prevalence of the late-phase hyperfluorescence was statistically significant in early age-related maculopathy. Eight of the 22 eyes with early age-related maculopathy and one of the five eyes with normal aging progressed to exudative AMD. Conclusions: Hyperfluorescent areas observed in late-phase IA before the manifestation of exudative AMD suggested the possible presence of choroidal neovascular membrane in the subretinal pigment epithelial space.

Reduced amplitude and delayed latency in foveal response of multifocal electroretinogram in early age related macular degeneration

AIMTo explore the use of multifocal electroretinograms (MERG) in detecting early changes in age related macular degeneration (AMD).METHOD15 pre-AMD or early AMD eyes showing retinal drusen or irregular fundus pigmentation...