PEDF-derived peptide protects against Amyloid-ß toxicity in vitro and prevents retinal dysfunction in rats.

Amyloid-beta (Aß), a family of aggregation-prone and neurotoxic peptides, has been implicated in the pathophysiology of age-related macular degeneration (AMD). We have previously shown that oligomeric and fibrillar species of Aß42 exerted retinal toxicity in rats, but while the consequences of exposure to amyloid were related to intracellular effects, the mechanism of Aß42 internalization in the retina is not well characterized. In the brain, the 67 kDa laminin receptor (67LR) participates in Aß-related neuronal cell death. A short peptide derived from pigment epithelium-derived factor (PEDF), formerly designated PEDF-335, was found to mitigate experimental models of ischemic retinopathy via targeting of 67LR. In the present study, we hypothesized that 67LR mediates the uptake of pathogenic Aß42 assemblies in the retina, and that targeting of this receptor by PEDF-335 may limit the internalization of Aß, thereby ameliorating its retinotoxicity. To test this assumption ARPE-19 cells in culture were incubated with PEDF-335 before treatment with fibrillar or oligomeric structures of Aß42. Immunostaining confirmed that PEDF-335 treatment substantially prevented amyloid internalization into ARPE-19 cells and maintained their viability in the presence of toxic oligomeric and fibrillar Aß42 entities in vitro. FRET competition assay was performed and confirmed the binding of PEDF-335 to 67LR in RPE-like cells. Wild-type rats were treated with intravitreal PEDF-335 in the experimental eye 2 days prior to administration of retinotoxic Aß42 oligomers or fibrils to both eyes. Retinal function was assessed by electroretinography through 6 weeks post injection. The ERG responses in rats treated with oligomeric or fibrillar Aß42 assemblies were near-normal in eyes previously treated with intravitreal PEDF-335, whereas those measured in the control eyes treated with injection of the Aß42 assemblies alone showed pathologic attenuation of the retinal function through 6 weeks. The retinal presence of 67LR was determined ex vivo by immunostaining and western blotting. Retinal staining demonstrated the constitutional expression of 67LR mainly in the retinal nuclear layers. In the presence of Aß42, the levels of 67LR were increased, although its retinal distribution remained largely unaltered. In contrast, no apparent differences in the retinal expression level of 67LR were noted following exposure to PEDF-335 alone, and its pattern of localization in the retina remained similarly concentrated primarily in the inner and outer nuclear layers. In summary, we found that PEDF-335 confers protection against Aß42-mediated retinal toxicity, with significant effects noted in cells as well as in vivo in rats. The effects of PEDF-335 in the retina are potentially mediated via binding to 67LR and by at least partial inhibition of Aß42 internalization. These results suggest that PEDF-335 may merit further consideration in the development of targeted inhibition of amyloid-related toxicity in the retina. More broadly, our observations provide evidence on the importance of extracellular versus intracellular Aß42 in the retina and suggest concepts on the molecular mechanism of Aß retinal pathogenicity.

Changes in chorioretinal flow index after cataract surgery: an optical coherence tomography angiography study.

PURPOSE: Pseudophakic cystoid macular edema (CME) occurs in up to 2% of uneventful cataract surgeries. This study evaluates changes in macular blood flow succeeding uneventful phacoemulsification cataract extraction among otherwise visually healthy subjects. METHODS: This prospective study included 18 eyes of 18 patients undergoing routine phacoemulsification. Optical coherence tomography angiography (OCT-A) was performed using the Angio-Retina 6 × 6 mm protocol with the XR Avanti Angio-Vue system (Optovue Inc., Fremont, California) prior to the surgery and 4-8 weeks thereafter. Exclusion criteria included motion artifacts, segmentation errors and signal strength index (SSI) < 40. The main outcome measure was change in flow index (FI) measured in all 4 retinal segmentation layers within an area of 1 mm diameter around the foveal center. RESULTS: Following surgery, a significant increase in SSI (46.65 ± 8.62 versus 53.12 ± 8.07, p = 0.01), superficial plexus FI (0.98 ± 0.23 versus 1.16 ± 0.16, p = 0.02) and deep plexus FI (0.54 ± 0.46 versus 0.93 ± 0.39, p = 0.01) was found. No significant changes were noted in the outer retina or the choriocapillaris. CONCLUSION: The study demonstrates a significant increase in FI in the superficial and deep retinal plexus following uneventful cataract surgery, with the greatest changes occurring in the latter. These findings corroborate evidence from structural imaging and support the vascular etiology of pseudophakic CME.

Is there room for prompt routine ophthalmic examination in neonates with congenital cytomegalovirus (CMV) infection during the neonatal period?

PURPOSE: To establish the clinical necessity of routine targeted ophthalmic examination of newborns with congenital cytomegalovirus (CMV) infection during the neonatal period. METHODS: This retrospective study included consecutive neonates that were referred for ophthalmological screening within the context of a proven congenital CMV infection. The presence of CMV-related ocular and systemic findings was determined. RESULTS: Among the 91 patients included in this study, 72 (79.12%) were symptomatic with one or more of the following manifestations: abnormal brain ultrasound (42; 46.15%), small for gestational age (29; 31.87%), microcephaly (23; 25.27%), thrombocytopenia (14; 15.38%), sensory neural hearing loss (13; 14.29%), neutropenia (12; 13.19%), anemia (4; 4.4%), skin lesions (4; 4.4%), hepatomegaly (3; 3.3%), splenomegaly (3; 3.3%), direct hyperbilirubinemia (2; 2.2%). Not one single neonate in this cohort had any of the ocular findings surveyed. CONCLUSION: The presence of ophthalmological findings among neonates with congenital CMV infection during the neonatal period is infrequent, suggesting that routine ophthalmological screening may be safely deferred for the post-neonatal period.

Autosomal dominant retinitis pigmentosa with incomplete penetrance due to an intronic mutation of the PRPF31 gene.

Purpose: To identify the molecular mechanisms of the development of autosomal dominant retinitis pigmentosa (adRP) with incomplete penetrance in an Israeli Muslim Arab family. Methods: Two patients with adRP underwent a detailed ophthalmic evaluation, including funduscopic examination, visual field testing, optical coherence tomography, and electroretinography. Genetic analysis was performed using a combination of whole exome sequencing (WES) and Sanger sequencing. The pathogenicity of the identified intronic variant was evaluated in silico using several web-based tools, in vitro using a minigene-based assay, and in vivo using reverse transcription PCR analysis of lymphocyte-derived RNA. The relative abundance of alternatively spliced transcripts was evaluated using amplicon-based next-generation sequencing. The relative expression levels of PRPF31 and CNOT3 were measured using quantitative PCR (qPCR) analysis. Results: The two patients recruited in this study had childhood-onset RP, with night blindness as the initial symptom, followed by concentric restriction of the visual field. The funduscopic findings included narrowed retinal blood vessels and peripheral bone spicule pigmentation. By the third decade of life, the full-field electroretinography findings had been remarkably attenuated. In these patients, we identified a novel heterozygous intronic variant at position +5 of PRPF31 intron 11 (c.1146+5G>T). The same variant was also detected in one asymptomatic family member. Through in silico analysis, the variant was predicted to alter the splicing of intron 11. An in vitro splicing assay and a reverse transcription PCR analysis of lymphocyte-derived RNA revealed that the mutant allele yielded mainly a shorter transcript in which exon 11 was skipped. The skipping of exon 11 was expected to cause a frameshift and an aberrant truncated protein (p.Tyr359Serfs*29). The qPCR analysis revealed reduced PRPF31 expression levels in the mutation carriers, without a significant difference between the affected patient and his asymptomatic brother. We evaluated several factors that have been suggested to correlate with non-penetrance of PRPF31 mutations, including the number of cis-acting MSR1 elements adjacent to the PRPF31 core promoter, CNOT3 expression level, and CNOT3 rs4806718 single-nucleotide polymorphism. None of these factors correlated with non-penetrance in the family in this study. Conclusions: We report a novel intronic mutation in PRPF31 underlying adRP. This report expands the spectrum of pathogenic mutations in PRPF31 and further demonstrates the importance of intronic mutations. Moreover, it demonstrates the phenomenon of incomplete penetrance previously associated with PRPF31 mutations. The fact that the non-penetrance in the family in this study could not be explained by any of the known mechanisms suggests the possible contribution of a novel modifier of PRPF31 penetrance.

SELF-DESIGNATION OF THE TREATED EYE BEFORE INTRAVITREAL INJECTIONS: Prevalence and Predictors of Incorrect Calling.

PURPOSE: To identify risk factors for incorrect self-identification of the treatment eye before intravitreal injections. METHODS: This prospective study included consecutive patients who were asked to designate the eye for which the intravitreal injection was intended and were subsequently divided into two groups according to whether or not they identified the correct eye. RESULTS: Overall, 349 eyes (n = 349) were included, and 8.6% (n = 30) designated the incorrect eye or did not know which eye was intended for treatment. Incorrect designation was associated with diabetic macular edema (odds ratio [OR] = 0.33 [0.15-0.75]), first injection in the intended eye or ≥1 year since previous injection (OR = 0.34 [0.14-0.87]), Arabic native tongue (OR = 0.48 [0.22-1.01]), previous injection to the fellow eye (OR = 0.26 [0.10-0.64]), and concurrent treatment of both eyes (OR = 0.35 [0.16-0.74]). Multivariate analysis showed the first injection or ≥1 year since last injection in the treatment eye (R2 = 2.24%, P = 0.004, OR = 0.20 [0.07-0.57]) and previous injection in the fellow eye (R2 = 6.55%, P < 0.001, OR = 0.20 [0.07-0.52]) as significant independent predictors of incorrect identification. CONCLUSION: Several factors are associated with a greater probability for incorrect patient's self-identification of the eye laterality intended for intravitreal injections. These findings may help identify patients with a higher risk of such potential errors.

PERSISTENT LOCULATED SUBRETINAL FLUID AFTER RHEGMATOGENOUS RETINAL DETACHMENT SURGERY.

PURPOSE: To identify factors associated with persistent subretinal fluid (SRF) after small-gauge pars plana vitrectomy for primary rhegmatogenous retinal detachment. METHODS: This retrospective study included patients from 2 tertiary centers who underwent pars plana vitrectomy for repair of rhegmatogenous retinal detachment between 2013 and 2016. Preoperative and intraoperative parameters were examined for association with development of SRF. RESULTS: Overall, 153 eyes of 153 patients, mean age of 55.2 ± 17.9 years were included. Persistent SRF occurred in 15.0% (n = 23) and was associated with high myopia (65.22 vs. 26.15%, P < 0.001), macula-involving retinal detachment (91.30 vs. 66.15%, P = 0.02), phakic lens status (86.96 vs. 66.15%, P = 0.04), and younger age (47.8 ± 18.7 vs. 56.5 ± 17.5, P = 0.04) while drainage retinotomy was protective (13.04 vs. 34.11%, P = 0.04). In multivariate analysis, high myopia (P = 0.009) and macula-involving retinal detachment (P = 0.004) were associated with SRF, while drainage retinotomy was protective (P = 0.03). Persistent SRF was associated with outer retinal band irregularity (30.4 vs. 9.3%, P = 0.005). There were no significant differences in terms of change in best-corrected visual acuity from presentation (P = 0.70), or final best-corrected visual acuity (P = 0.54). CONCLUSION: Eyes with preoperative high myopia and macular involvement, and those in which a drainage retinotomy was not performed, were more likely to develop persistent SRF.

Infliximab exerts a dose-dependent effect on retinal safety in the albino rabbit.

PURPOSE: To assess the retinal toxicity of an intravitreal injection of infliximab, a monoclonal antibody to tumor necrosis factor α, in a rabbit model. MATERIALS AND METHODS: Two groups of adult albino rabbits (n = 5) received intravitreal injections of infliximab (0.1 ml) in the study eye and balanced salt solution (BSS, 0.1 ml) in the control eye at baseline. Group 1 was administered with 1.5 mg/0.1 ml, and group 2 was injected with 7.5 mg/0.1 ml of infliximab solution. Electroretinography (ERG) was performed at baseline and at 1, 7, 30, and 45 days after the injection. Visual evoked potentials (VEPs) were recorded at 7 and 45 days after the injection. After the last electrophysiological assessment, the rabbits were euthanized and retinal histopathology and immunhistochemistry for glial fibrillary acidic protein (GFAP) were performed. RESULTS: ERG responses demonstrated no significant deficit in retinal function in eyes injected with infliximab. Mean dark-adapted a-wave and b-wave maximal amplitude and semi-saturation constant values at baseline and throughout the 45 days of follow-up after the injection indicated no remarkable difference in outer retinal function between the control and experimental eyes. VEP responses were similar at each time point (7 and 45 days). No difference was seen in retinal histopathology and immunocytochemistry sections in eyes receiving the 1.5 mg/0.1 ml dose compared to the control eyes. However, increased GFAP labeling in retinal Müller cells was detected in rabbit eyes treated with the 7.5 mg/0.1 ml dose. CONCLUSIONS: Intravitreal injection of 1.5 mg/0.1 ml infliximab dose has no toxic effect on the integrity (functional or structural) of the retina in rabbits. A higher dose of 7.5 mg/0.1 ml may be slightly toxic as suggested by positive Müller cell GFAP expression. Additional studies of retinal toxicity at higher doses and after multiple injections are needed to establish the retinal safety of intravitreal infliximab therapy in humans.

Trends in Authorship of Articles in Major Ophthalmology Journals by Gender, 2002-2014.

PURPOSE: To evaluate trends in the prevalence of women authors in ophthalmology in recent years. DESIGN: Cohort study. PARTICIPANTS: Authors listed in publications of 6 leading ophthalmology journals between January 2002 and December 2014. METHODS: Using the PubMed search engine, we conducted an observational study of trends in gender distribution of all authors in 6 leading ophthalmology journals between January 2002 and December 2014. In multiauthored articles, the first listed author often is the lead investigator and the last author is the senior author. Therefore, the full names and positions (first, middle, or last) of all authors in every article were collected. A Google-based name identifier was used to assign the gender of authors. MAIN OUTCOME MEASURES: Proportion of women authors throughout the study period in all journals, general ophthalmology versus subspecialty journals, and basic science versus clinical research journals. Furthermore, we assessed the proportion of women in different authorship positions (first, middle, and last). RESULTS: A total of 102 254 authors from 23 026 published articles were analyzed. There was a significant rise over time in the percentage of women authors, with a steeper slope for first authors than for last authors (P<0.001), although in 2014, women authors were less than the 50% mark in all categories of authorship. The rise in the percentage of women authors was similar in basic and clinical research, but was steeper for first authorship than for last authorship (P<0.001). In all 3 authorship positions (first, middle, or last), women's contributions consistently were higher in basic research publications. The rise in the percentage of women authors was significantly steeper for general journals than for subspecialty journals (P<0.001). There was no significant rise for last authorship in subspecialty journals. In all 3 authorship positions, the proportion of women was consistently higher in general ophthalmology journals than for subspecialty journals. CONCLUSIONS: Despite an overall increase in the contribution of women to the field of ophthalmology, contributions to articles published in subspecialty ophthalmology journals and the proportion of women listed as last authors on overall articles published in ophthalmology journals are still low.

The Surprising Nonlinear Effects of S100A9 Proteins in the Retina.

Age-related macular degeneration (AMD) is a complex disease in which inflammation is implicated as a key factor but the precise molecular mechanisms are poorly understood. AMD lesions contain an excess of the pro-inflammatory S100A9 protein, but its retinal significance was yet unexplored. S100A9 was shown to be intrinsically amyloidogenic in vitro and in vivo. Here, we hypothesized that the retinal effects of S100A9 are related to its supramolecular conformation. ARPE-19 cultures were treated with native dimeric and fibrillar S100A9 preparations, and cell viability was determined. Wild-type rats were treated intravitreally with the S100A9 solutions in the right eye and with the vehicle in the left. Retinal function was assessed longitudinally by electroretinography (ERG), comparing the amplitudes and configurations for each intervention. Native S100A9 had no impact on cellular viability in vitro or on the retinal function in vivo. Despite dispersed intracellular uptake, fibrillar S100A9 did not decrease ARPE-19 cell viability. In contrast, S100A9 fibrils impaired retinal function in vivo following intravitreal injection in rats. Intriguingly, low-dose fibrillar S100A9 induced contrasting in vivo effects, significantly increasing the ERG responses, particularly over 14 days postinjection. The retinal effects of S100A9 were further characterized by glial and microglial cell activation. We provide the first indication for the retinal effects of S100A9, showing that its fibrils inflicted retinal dysfunction and glial activation in vivo, while low dose of the same assemblies resulted in an unpredicted enhancement of the ERG amplitudes. These nonlinear responses highlight the consequences of self-assembly of S100A9 and provide insight into its pathophysiological and possibly physiological roles in the retina.

Reduced Electroretinogram Responses in Morphologically Normal Retina in Patients with Primary Hyperoxaluria Type 1.

Purpose: To describe ocular findings in individuals with primary hyperoxaluria type 1 (PH1), focusing on the correlations between retinal anatomy and retinal function. To characterize the retinal alterations that occur at different disease stages by evaluating individuals with diverse degrees of renal impairment associated with PH1. Design: A cross-sectional study. Participants: Patients diagnosed with PH1 based on clinical criteria and genetic testing, treated in the Pediatric Nephrology Unit of the Ruth Children's Hospital, Rambam Health Care Campus, Haifa, Israel between 2013 and 2021. Methods: The ophthalmological assessment included a slit-lamp biomicroscopy of the anterior and posterior segment or indirect ophthalmoscopy. Electroretinography was employed for assessment of the retinal function, and retinal imaging included spectral-domain OCT and fundus autofluorescence. A systematic evaluation of the disease stage was based on clinical criteria including physical examination, purposeful imaging (X-ray, echocardiography, and US abdomen), and laboratory tests as needed. Main Outcome Measures: Anatomical and functional assessment of the retina in patients with PH1, and the relationship between retinal dysfunction and kidney impairment. Results: A total of 16 eyes were examined in the study of 8 children ranging in age from 4 to 19 years. Four eyes (25%) showed normal structural and functional retinal findings, 8 eyes (50%) presented functional impairment in the absence of pathological structural findings, and 4 eyes (25%) had advanced retinal damage that manifested as significant morphological and functional impairment. There was no direct relationship between the severity of the renal disease and the severity of the retinal phenotype. Conclusions: Subjects with PH1 present varying severity levels of the retinal phenotype, with possible discrepancy between the clinical retinal morphology and the retinal function noted on electroretinography. These findings raise questions about the molecular basis of the retinal manifestations in PH1. The presence of functional impairment in the absence of evident crystal deposition in the retina suggests that, in addition to oxalate crystal accumulation, other biomolecular processes may play a role in the development of retinopathy.

Safety of intravitreal bevacizumab in the developing rabbit retina.

PURPOSE: The purpose of this was to study the long-term effects of a single intravitreal dose of bevacizumab injected at different postnatal age on retinal function and development of retinal blood vessels in the adult albino rabbit. METHODS: Bevacizumab was injected into the right eye of newborn rabbits, aged 11 days to 25 days, whereas the left eye of each rabbit was injected with identical volume of saline and served as control. The electroretinogram was recorded 1 week and 10 weeks after injection. Visual evoked potentials were recorded 10 weeks after injection. At termination of the follow-up period, the rabbits were killed and the retinas were prepared for histopathologic studies at the light microscopic level, for glial fibrillary acidic protein immunoreactivity, and for reduced form of nicotinamide adenine dinucleotide phosphate diaphorase histochemistry to assess the integrity of the retinal vascular system. RESULTS: The electroretinogram responses demonstrated normal retinal function in adult rabbits injected at postnatal age of 11 days to 25 days. Mean Vmax and σ values calculated for each group of rabbits, 10 weeks after bevacizumab injection, indicated similar retinal function of the experimental and control eyes. Visual evoked potentials recorded by stimulating each eye separately were also similar. The histopathologic studies yielded similar results; no structural retinal damage was observed in the experimental eyes compared with the control eyes of rabbits from all age groups, and no increased glial fibrillary acidic protein immunoreactivity in Müller cells was observed in the experimental eyes. Staining of blood vessels with reduced form of nicotinamide adenine dinucleotide phosphate diaphorase revealed decreased branching of the capillary network in the experimental eyes compared with the control eyes in all age groups. CONCLUSION: The electroretinographic and morphologic data showed no deleterious effects of a single intravitreal dose of bevacizumab, injected during the first 30 days postnatally, on the structural and functional integrity of the sensory retina in the adult rabbit. Even partial blockage of vascular endothelial growth factor with bevacizumab applied during retinal development seems to interfere with the development of the retinal vascular system in the albino rabbit. However, extrapolation from rabbits to humans should be made with caution.

Predicting Post-Cataract Surgery Visual Acuity in Vitrectomized Eyes: The Efficacy and Accuracy of "Lambda" Retinometry.

BACKGROUND AND OBJECTIVE: Determine the ability of Lambda retinometry to predict post-cataract surgery visual acuity in vitrectomized eyes. PATIENTS AND METHODS: Prospective study including 47 cataract surgery candidates with a history of pars plana vitrectomy (PPV). Lambda retinometry using a hand-held Lambda retinometer and best-corrected visual acuity (BCVA) were measured preoperatively, and BCVA was reassessed postoperatively. RESULTS: Lambda predictions strongly correlated with postoperative BCVA (logarithm of the minimum angle of resolution [logMAR]) (P < .001, r2 = 0.57), especially combined with preoperative BCVA (logMAR) (P < .001, r2 = 0.65). In 89% of cases, postoperative BCVA was equal to or higher than the prediction. Neither cataract grades nor indications for PPV were associated with the accuracy of Lambda predictions (P = .882 and P = .790, respectively). Underestimation of visual outcome was more common than overestimation. A Lambda prediction of ≥ 20/40 (Snellen) had a positive predictive value of 85.7% and a negative predictive value of 73.6% for the postoperative outcome. CONCLUSIONS: Lambda retinometry can reliably predict the postoperative BCVA in cataract patients who previously underwent PPV, with a tendency towards underestimation. [Ophthalmic Surg Lasers Imaging Retina. 2021;52:535-542.].

Inhibition of amyloid fibrillation of γD-crystallin model peptide by the cochineal Carmine.

γD-crystallin is among the most abundant γ-crystallins in the human eye lens which are essential for preserving its transparency. Aging, and environmental changes, cause crystallins to lose their native soluble structure and aggregate, resulting in the formation of cataract. Current treatment of cataract is surgical removal which is costly. Pharmaceutical therapeutics of cataract is an unmet need. We report a screen for small molecules capable of inhibiting aggregation of human γD-crystallin. Using a highly amyloidogenic hexapeptide model 41GCWMLY46 derived from the full-length protein, we screened a library of 68 anthraquinone molecules using ThT fluorescence assay. A leading hit, the cochineal Carmine, effectively reduced aggregation of the model GDC6 peptide in dose dependent manner. Similar effect was observed toward aggregation of the full-length γD-crystallin. Transmission electron microscopy, intrinsic Tryptophan fluorescence and ANS fluorescence assays corroborated these results. Insights obtained from molecular docking suggested that Carmine interaction with monomeric GDC6 involved hydrogen bonding with Ace group, Cys, Met residues and hydrophobic contact with Trp residue. Carmine was non-toxic toward retinal cells in culture. It also reduced ex vivo the turbidity of human extracted cataract material. Collectively, our results indicate that Carmine could be used for developing new therapeutics to treat cataract.

Safety evaluation of repeated intravitreal injections of bevacizumab and ranibizumab in rabbit eyes.

PURPOSE: Repeated intravitreal injections of ranibizumab or bevacizumab are a common treatment for several retinal diseases. The aim of the present study was to evaluate the long-term retinal toxicity of repeated injections of ranibizumab and bevacizumab in rabbits. METHODS: Albino rabbits were injected intravitreally with ranibizumab (1 mg/0.1 mL) or bevacizumab (2.5 mg/0.1 mL) into the right eye, whereas the left eye of each rabbit was injected with saline. Nine consecutive injections were administered at 14-day intervals. Electroretinographic responses and flash visual-evoked potentials were recorded periodically. After 18 weeks of follow-up, the rabbits were killed, and the retinas were prepared for morphologic examination and for immunocytochemistry for glial fibrillary acidic protein. RESULTS: Electroretinographic and visual-evoked potential responses of the experimental and control eyes were similar in amplitude and pattern throughout the follow-up period. The histopathologic studies yielded similar results. No retinal damage was observed in the experimental and control eyes of all rabbits. Glial fibrillary acidic protein immunoreactivity showed staining only in retinal astrocytes but not in Müller cells in all rabbits. CONCLUSION: The electrophysiological tests and the morphologic data indicate that the repeated intravitreal injections of ranibizumab or bevacizumab have no cumulative long-term toxic effect on the retina in rabbits.

Clinical characteristics of endophthalmitis after an injection of intravitreal antivascular endothelial growth factor.

PURPOSE: The purpose of this study was to describe the clinical characteristics and management of patients with bacterial endophthalmitis after an intravitreal antivascular endothelial growth factor injection. METHODS: This is a retrospective chart review of all patients admitted with suspected endophthalmitis from 2006 to 2008. RESULTS: Endophthalmitis was verified by positive Gram stain and culture in nine eyes. The mean preinjection visual acuity of the 9 eyes was 0.02 +/- 0.021 diopters (decimal visual acuity scale) and dropped to 0.01667 +/- 0.02449 diopters in the eyes with endophthalmitis. All nine patients presented with reduced visual acuity, of whom seven also had ocular pain. Initial treatment was administered without delay and consisted of vitreous tap and intravitreal antibiotics injection in five cases and pars plana vitrectomy with intravitreal antibiotics injection in the other four cases. Vitreous tap failed in one case. Seven patients underwent a second procedure and two underwent a third procedure. The mean posttreatment visual acuity in all 9 eyes improved significantly (0.19 +/- 0.24, P = 0.0071). Five patients had major complications (e.g., retinal detachment, phacolytic glaucoma, and recurrent endophthalmitis). CONCLUSION: Acute endophthalmitis following anti-VEGF injection appears within days and can result in severe loss of vision if not treated promptly. In our series the clinical and prognostic characteristics were considerably different between culture positive endophthalmitis cases and culture negative cases.

The effect of contact lens wear on retinal spectral domain optical coherence tomography.

BACKGROUND: This study assessed the impact of contact lens wear on retinal spectral domain optical coherence tomography (SD-OCT) image quality and macular thickness measurements, among subjects with myopia. METHODS: This was a prospective study including 34 subjects (26.59 ± 3.19 years) with myopia or myopic astigmatism. Twelve were imaged wearing spherical soft contact lenses, eight non-contact lens wearers were imaged with a plano soft contact lens, and 14 with significant astigmatism were fitted with a rigid gas permeable (RGP) contact lens. For each group of contact lens types, the average image quality index (Q-index), and the average macular thickness measurements were compared between macular OCT scans obtained from the same eyes with and without a contact lens. RESULTS: Among the subjects assessed with their habitual spherical soft lenses, the average Q-index was similar for scans acquired with and without a contact lens (30.10 ± 1.94 versus 31.03 ± 2.55; p = 0.18). Among non-contact lens wearers, the average Q-index was slightly higher for scans acquired without a contact lens, compared to scans with a plano contact lens (31.99 ± 2.06 versus 29.51 ± 1.56; p = 0.006). Among 14 subjects imaged wearing a fitted RGP contact lens, the Q-index was similar for scans acquired with and without a contact lens (29.04 ± 2.73 versus 28.75 ± 2.86; p = 0.78). In all groups, there were no correlations between the power of the sphere and change in the Q-index (that is, post- minus pre-contact lens Q-index), and no differences were found between OCT-derived macular thickness measurements from scans with and without a contact lens. The magnitude of cylinder was not correlated with the change in the Q-index in the habitual and RGP contact lens groups. However, an inverse correlation between cylinder power and change in the Q-index was found in the plano contact lens group. CONCLUSION: In low to intermediate levels of myopia, with or without regular astigmatism, macular SD-OCT imaging does not merit placement of a soft or rigid contact lens, nor is there an added benefit from removing a habitual spherical soft lens prior to scanning.

Preoperative Anxiety Levels and Pain during Cataract Surgery.

Purpose: To evaluate the correlation between pain during cataract surgery and preoperative anxiety.Methods: This prospective observational masked study included 103 eyes of 103 consecutive patients who underwent routine clear corneal incision phacoemulsification surgery at the Department of Ophthalmology, Rambam Health Care Campus, Haifa, Israel. Patients were interviewed prior to surgery and 5 min following surgery by two separate independent interviewers. Anxiety level was measured by the Visual Analog Scale for Anxiety (VASA) and pain by the Visual Analog Scale (VAS). The main outcome measure was the maximum amount of pain endured during the procedure using VAS.Results: The mean participant age was 68.9 ± 8.9 years, and 46.6% were male. Severe anxiety (VASA ≥ 7) and pain (VAS ≥ 7) were documented in 18.5 and 17.5% of patients, respectively. There was a statistically significant positive correlation between VAS and VASA (r = 0.62, p < .001) as well as between VAS and duration of surgery (r = 0.20, p = .04). There was no association between VAS and all other investigated parameters in the univariate analysis. In backward regression analysis, VASA was the only parameter that was significantly associated with VAS (R2 = 36.61%, p < .001). Patients with severe anxiety were >10 times more likely to experience severe pain, and a VASA > 4 predicted severe pain with a sensitivity of 88.9% and a specificity of 69.4%.Conclusions: One-fifth of patients experienced severe anxiety and pain. Preoperative anxiety levels were the only significant predictor of pain. Reducing preoperative anxiety in cataract patients is warranted.

Induction of retinopathy by fibrillar oxalate assemblies.

The formation of metabolite fibrillar assemblies represents a paradigm shift in the study of human metabolic disorders. Yet, direct clinical relevance has been attributed only to metabolite crystals. A notable example for metabolite crystallization is calcium oxalate crystals observed in various diseases, including primary hyperoxaluria. We unexpectedly observed retinal damage among young hyperoxaluria patients in the absence of crystals. Exploring the possible formation of alternative supramolecular organizations and their biological role, here we show that oxalate can form ordered fibrils with no associated calcium. These fibrils inflict intense retinal cytotoxicity in cultured cells. A rat model injected with oxalate fibrils recaptures patterns of retinal dysfunction observed in patients. Antibodies purified from hyperoxaluria patient sera recognize oxalate fibrils regardless of the presence of calcium. These findings highlight a new molecular basis for oxalate-associated disease, and to our knowledge provide the first direct clinical indication for the pathogenic role of metabolite fibrillar assemblies.

The retinal toxicity profile towards assemblies of Amyloid-ß indicate the predominant pathophysiological activity of oligomeric species.

Amyloid-ß (Aß), reported as a significant constituent of drusen, was implicated in the pathophysiology of age-related macular degeneration (AMD), yet the identity of the major pathogenic Aß species in the retina has remained hitherto unclear. Here, we examined the in-vivo retinal impact of distinct supramolecular assemblies of Aß. Fibrillar (Aß40, Aß42) and oligomeric (Aß42) preparations showed clear biophysical hallmarks of amyloid assemblies. Measures of retinal structure and function were studied longitudinally following intravitreal administration of the various Aß assemblies in rats. Electroretinography (ERG) delineated differential retinal neurotoxicity of Aß species. Oligomeric Aß42 inflicted the major toxic effect, exerting diminished ERG responses through 30 days post injection. A lesser degree of retinal dysfunction was noted following treatment with fibrillar Aß42, whereas no retinal compromise was recorded in response to Aß40 fibrils. The toxic effect of Aß42 architectures was further reflected by retinal glial response. Fluorescence labelling of Aß42 species was used to detect their accumulation into the retinal tissue. These results provide conceptual evidence of the differential toxicity of particular Aß species in-vivo, and promote the mechanistic understanding of their retinal pathogenicity. Stratifying the impact of pathological Aß aggregation in the retina may merit further investigation to decipher the pathophysiological relevance of processes of molecular self-assembly in retinal disorders.

Cone Spacing Correlates With Retinal Thickness and Microperimetry in Patients With Inherited Retinal Degenerations.

Purpose: To determine whether high-resolution retinal imaging measures of macular structure correlate with visual function over 36 months in retinal degeneration (RD) patients and normal subjects. Methods: Twenty-six eyes of 16 RD patients and 16 eyes of 8 normal subjects were studied at baseline; 15 eyes (14 RD) and 11 eyes (6 normal) were studied 36 months later. Adaptive Optics Scanning Laser Ophthalmoscopy (AOSLO) was used to identify regions of interest (ROIs) with unambiguous cones at baseline to measure cone spacing. AOSLO images were aligned with spectral-domain optical coherence tomography (SD-OCT) and fundus-guided microperimetry results to correlate structure and function at the ROIs. SD-OCT images were segmented to measure inner segment (IS) and outer segment (OS) thickness. Correlations between cone spacing, IS and OS thickness and sensitivity were assessed using Spearman correlation coefficient ρ with bootstrap analyses clustered by person. Results: Cone spacing (ρ = 0.57, P < 0.001) and macular sensitivity (ρ = 0.19, P = 0.14) were significantly correlated with eccentricity in patients. Controlling for eccentricity, cone spacing Z-scores were inversely correlated with IS (ρ = -0.29, P = 0.002) and OS thickness (ρ = -0.39, P < 0.001) in RD patients only, and with sensitivity in normal subjects (ρ = -0.22, P < 0.001) and RD patients (ρ = -0.38, P < 0.001). After 36 months, cone spacing increased (P < 0.001) and macular sensitivity decreased (P = 0.007) compared to baseline in RD patients. Conclusions: Cone spacing increased and macular sensitivity declined significantly in RD patients over 36 months. High resolution images of cone structure correlated with retinal sensitivity, and may be appropriate outcome measures for clinical trials in RD.