Morphological and Clinical Characterization of Foveal Bulge Sign Three Years After Retinal Detachment Repair: A Longitudinal Prospective Evaluation.

Purpose: To evaluate optical-coherence-tomography (OCT)-morphological and clinical parameters three years after primary rhegmatogenous retinal detachment (RRD) repair surgery and the role of postoperative foveal bulge sign. Methods: Of the 32 initially enrolled patients with primary RRD 20 (14 fovea-on, 6 fovea-off) completed this prospective study. Preoperatively and 3 years after surgery best corrected visual acuity (BCVA) and OCT including macula status, central retinal thickness (CRT), central choroidal thickness (CCT), thickness of each single retinal layer, integrity of cone-interdigitation-zone (CIZ) and ellipsoid zone (EZ), and presence of foveal bulge were evaluated. Results: Preoperatively fovea-off RRD patients show significantly thinner CCT, inner nuclear layer (INL) and inner plexiform layer (IPL) compared to fovea-on RRD patients, whereas only IPL and INL were significantly thicker compared to the fellow eye. Three years after surgery this thickening recovered. Final BCVA did not differ statistically significantly between fovea-off and fovea-on patients, no difference in CIZ-integrity or presence of foveal bulge was observed. Presence of foveal bulge at 3 years showed significantly better final BCVA and was associated with intact preoperative CIZ-integrity and postoperative EZ- and CIZ-integrity. The preoperative fovea status showed no correlation to the postoperative presence of foveal bulge. Conclusion: Three years after RRD repair retinal layers show similar thickness. The presence of foveal bulge is associated with better final BCVA. Sufficient pre- and postoperative CIZ-integrity as well as postoperative CIZ-integrity seem to be strongly associated with the restoration of foveal bulge. No correlation was found with the preoperative macular status or BCVA.

We analyzed long-term optical-coherence-tomography changes after rhegmatogenous retinal detachment repair. Three years after rhegmatogenous retinal detachment repair retinal layers show similar thickness and visual acuity did not differ between fovea off and fovea on patients. The restoration of foveal bulge sign was the most important factor for good postoperative visual acuity. Integrity of specific retinal layers, such as the ellipsoid zone and cone-interdigitation-zone, seems to be strongly associated with the restoration of the foveal bulge.

A mega-analysis of expression quantitative trait loci in retinal tissue.

Significant association signals from genome-wide association studies (GWAS) point to genomic regions of interest. However, for most loci the causative genetic variant remains undefined. Determining expression quantitative trait loci (eQTL) in a disease relevant tissue is an excellent approach to zoom in on disease- or trait-associated association signals and hitherto on relevant disease mechanisms. To this end, we explored regulation of gene expression in healthy retina (n = 311) and generated the largest cis-eQTL data set available to date. Genotype- and RNA-Seq data underwent rigorous quality control protocols before FastQTL was applied to assess the influence of genetic markers on local (cis) gene expression. Our analysis identified 403,151 significant eQTL variants (eVariants) that regulate 3,007 genes (eGenes) (Q-Value < 0.05). A conditional analysis revealed 744 independent secondary eQTL signals for 598 of the 3,007 eGenes. Interestingly, 99,165 (24.71%) of all unique eVariants regulate the expression of more than one eGene. Filtering the dataset for eVariants regulating three or more eGenes revealed 96 potential regulatory clusters. Of these, 31 harbour 130 genes which are partially regulated by the same genetic signal. To correlate eQTL and association signals, GWAS data from twelve complex eye diseases or traits were included and resulted in identification of 80 eGenes with potential association. Remarkably, expression of 10 genes is regulated by eVariants associated with multiple eye diseases or traits. In conclusion, we generated a unique catalogue of gene expression regulation in healthy retinal tissue and applied this resource to identify potentially pleiotropic effects in highly prevalent human eye diseases. Our study provides an excellent basis to further explore mechanisms of various retinal disease etiologies.

Comparing Different Decontamination Procedures in Harvesting Human Donor Cornea.

PURPOSE: To evaluate the effects of current hygiene standards for the enucleation of postmortem eyes by investigating the number of microorganisms during subsequent steps of decontamination and tissue processing. MATERIALS AND METHODS: This prospective, non-randomized cohort study includes 184 postmortem eyes of 92 human donors. Enucleation was performed, according to an ophthalmic surgical procedure. Two groups were generated as follows: right eyes were allocated to group A, left eyes were allocated to group B. In group A, a mucosal disinfectant was used accessorily. Conjunctival smears were taken to examine germ load in both groups before any intervention, in group A after mucosal disinfection, in both groups after transportation of the whole globes in transport fluid, and in both groups after a bath in 0.75% povidone iodine solution for at least 3 minutes just before preparation of the corneoscleral disc. Smears were sent to the local microbiologic laboratory in an aseptic package for testing. RESULTS: All smears showed multiple contaminations (n = 184/184 eyes) before treatment with povidone iodine solution. Contamination was in both groups significantly prevented using the treatment strategy of an at least 3-minute bath in 0.75% povidone iodine solution (n = 1/184 eyes; p < 0.01) As a side effect, almost every eye of group A and none of group B showed brown iodine staining corresponding to corneal epithelial erosion. CONCLUSIONS: An aseptic setting for donor enucleation similar to a surgical procedure seems not to influence the outcome of germ colonization. The most effective step to decontaminate donor eyes is to use 0.75% povidone iodine solution for at least 3 minutes.

Idebenone Prevents Oxidative Stress, Cell Death and Senescence of Retinal Pigment Epithelium Cells by Stabilizing BAX/Bcl-2 Ratio.

PURPOSE: Age-related macular degeneration (AMD) is one of the leading causes of blindness. Degeneration of the retinal pigment epithelium (RPE) is pathognomonic for the disease, and oxidative stress plays an important role in the pathogenesis of this disease. This study investigates potential antiapoptotic and cytoprotective effects of idebenone on cultured RPE cells (ARPE-19) under conditions of oxidative stress. METHODS: ARPE-19 cells were treated with 1-100 µM idebenone. Cell viability (MTT assay), induction of intracellular reactive oxygen species (ROS) and histone-associated DNA fragments in mono- and oligonucleosomes, expression of proapoptotic BAX and antiapoptotic Bcl-2 as well as senescence-associated ß-galactosidase (SA-ß-Gal) activity were investigated under exposure to hydrogen peroxide (H2O2). RESULTS: Idebenone concentrations from 1 to 20 µM showed no toxic effects on ARPE-19 cells. When cells were treated with H2O2, pretreatment with 5, 7.5, 10, and 20 µM idebenone led to a significant increase in the viability of ARPE-19 cells. In addition, idebenone pretreatment significantly attenuated the induction of SA-ß-Gal and intracellular ROS as well as the amount of histone-associated DNA fragments after treatment with H2O2. The reduction of proapoptotic BAX and the elevation of antiapoptotic Bcl-2 under idebenone show that this process is rather mediated by inhibiting H2O2-induced apoptosis, not necrosis. CONCLUSION: In this study, idebenone increased survival of ARPE-19 cells and reduced cell death, senescence, and oxidative stress by stabilizing the BAX/Bcl-2 ratio.

A Candidate Gene Association Study Identifies DAPL1 as a Female-Specific Susceptibility Locus for Age-Related Macular Degeneration (AMD).

Age-related macular degeneration (AMD) is the leading cause of blindness among white caucasians over the age of 50 years with a prevalence rate expected to increase markedly with an anticipated increase in the life span of the world population. To further expand our knowledge of the genetic architecture of the disease, we pursued a candidate gene approach assessing 25 genes and a total of 109 variants. Of these, synonymous single nucleotide polymorphism (SNP) rs17810398 located in death-associated protein-like 1 (DAPL1) was found to be associated with AMD in a joint analysis of 3,229 cases and 2,835 controls from five studies [combined PADJ = 1.15 × 10(-6), OR 1.332 (1.187-1.496)]. This association was characterized by a highly significant sex difference (Pdiff = 0.0032) in that it was clearly confined to females with genome-wide significance [PADJ = 2.62 × 10(-8), OR 1.541 (1.324-1.796); males: PADJ = 0.382, OR 1.084 (0.905-1.298)]. By targeted resequencing of risk and non-risk associated haplotypes in the DAPL1 locus, we identified additional potentially functional risk variants, namely a common 897-bp deletion and a SNP predicted to affect a putative binding site of an exonic splicing enhancer. We show that the risk haplotype correlates with a reduced retinal transcript level of two, less frequent, non-canonical DAPL1 isoforms. DAPL1 plays a role in epithelial differentiation and may be involved in apoptotic processes thereby suggesting a possible novel pathway in AMD pathogenesis.

Darbepoetin alpha reduces oxidative stress and chronic inflammation in atherosclerotic lesions of apo E deficient mice in experimental renal failure.

BACKGROUND: Cardiovascular morbidity and mortality is very important in patients with chronic renal failure. This occurs even in mild impairment of renal function and may be related to oxidative stress and chronic inflammation. The nephrectomized apo E knockout mouse is an accepted model for evaluating atherosclerosis in renal dysfunction. Erythropoietin derivates showed anti-oxidative and anti-inflammatory effects. Therefore, this study evaluates the effects of Darbepoetin on markers of oxidative stress and chronic inflammation in atherosclerotic lesions in apo E knockout mice with renal dysfunction. METHODS: Apo E knockout mice underwent unilateral (Unx, n = 20) or subtotal (Snx, n = 26) nephrectomy or sham operation (Sham, n = 16). Mice of each group were either treated with Darbepoetin or saline solution, a part of Snx mice received a tenfold higher dose of Darbepoetin. The aortic plaques were measured and morphologically characterized. Additional immunhistochemical analyses were performed on tissue samples taken from the heart and the aorta. RESULTS: Both Unx and Snx mice showed increased expression of markers of oxidative stress and chronic inflammation. While aortic plaque size was not different, Snx mice showed advanced plaque stages when compared to Unx mice. Darbepoetin treatment elevated hematocrit and lowered Nitrotyrosin as one marker of oxidative stress, inflammation in heart and aorta, plaque stage and in the high dose even plaque cholesterol content. In contrast, there was no influence of Darbepoetin on aortic plaque size; high dose Darbepoetin treatment resulted in elevated renal serum parameters. CONCLUSION: Darbepoetin showed some protective cardiovascular effects irrespective of renal function, i.e. it improved plaque structure and reduced some signs of oxidative stress and chronic inflammation without affecting plaque size. Nevertheless, the dose dependent adverse effects must be considered as high Darbepoetin treatment elevated serum urea. Elevation of hematocrit might be a favorable effect in anemic Snx animals but a thrombogenic risk in Sham animals.

Idebenone prevents human optic nerve head astrocytes from oxidative stress, apoptosis, and senescence by stabilizing BAX/Bcl-2 ratio.

PURPOSE: Oxidative stress plays an important role in the pathogenesis of several neurodegenerative diseases including glaucoma. Astrocytes are supposed to play a role in glaucoma pathogenesis. This study investigates the antiapoptotic and cytoprotective effects of idebenone on optic nerve head astrocytes (ONHA) under oxidative stress. METHODS: ONHA were treated with 1 to 150 µM idebenone. Cell viability (MTT assay and live-dead assay), induction of intracellular reactive oxygen species, senescence-associated ß-galactosidase activity were investigated. In addition, apoptosis (detection of histone-associated DNA fragmentation), and expression of BAX and Bcl-2, and their mRNA were determined after 48 hours and after hydrogen peroxide (H2O2) treatment. RESULTS: Idebenone concentrations from 1 to 50 µM showed no effects on ONHA viability. Pretreatment with 10 µM idebenone led to an increase in viability of ONHA after H2O2 treatment. In addition, idebenone pretreatment significantly attenuated the increase of histone-associated DNA fragmentation, induction of senescence-associated ß-galactosidase, and intracellular reactive oxygen species after treatment with H2O2. When ONHA cells were treated with idebenone and H2O2, real-time polymerase chain reaction and Western blot analysis yielded an increased expression of Bcl-2 and a decrease of BAX compared with those cells that were treated with H2O2 only. CONCLUSIONS: Idebenone reduced senescence, oxidative stress, and apoptotic cell death in cultured ONHA in vitro. Our results suggest that idebenone may help to protect ONHA in vivo, and therefore might be helpful in preventing the progression of glaucomatous degeneration.