A novel mutation of BEST1 gene in Best disease.

PURPOSE: To describe a new genetic variation of BEST1 gene in Best vitelliform macular dystrophy. METHODS: A patient with bilateral multiple retinal yellowish lesions at the posterior pole underwent fluorescein angiography, fundus autofluorescence, optical coherence tomography, electrooculogram and blood sample for genetic testing. RESULTS: A diagnosis of a Best vitelliform macular dystrophy was made. Heterozygous mutation c.76G > A (p.Gly26Ser) in exon 2 of the BEST1 gene was found. CONCLUSION: These findings contribute to expand the mutation spectrum of BEST1 gene.

Application of Artificial Intelligence in Targeting Retinal Diseases.

Retinal diseases affect an increasing number of patients worldwide because of the aging population. Request for diagnostic imaging in ophthalmology is ramping up, while the number of specialists keeps shrinking. Cutting-edge technology embedding artificial intelligence (AI) algorithms are thus advocated to help ophthalmologists perform their clinical tasks as well as to provide a source for the advancement of novel biomarkers. In particular, optical coherence tomography (OCT) evaluation of the retina can be augmented by algorithms based on machine learning and deep learning to early detect, qualitatively localize and quantitatively measure epi/intra/subretinal abnormalities or pathological features of macular or neural diseases. In this paper, we discuss the use of AI to facilitate efficacy and accuracy of retinal imaging in those diseases increasingly treated by intravitreal vascular endothelial growth factor (VEGF) inhibitors (i.e. anti-VEGF drugs), also including integration and interpretation features in the process. We review recent advances by AI in diabetic retinopathy, age-related macular degeneration, and retinopathy of prematurity that envision a potentially key role of highly automated systems in screening, early diagnosis, grading and individualized therapy. We discuss benefits and critical aspects of automating the evaluation of disease activity, recurrences, the timing of retreatment and therapeutically potential novel targets in ophthalmology. The impact of massive employment of AI to optimize clinical assistance and encourage tailored therapies for distinct patterns of retinal diseases is also discussed.

New Anti-VEGF Drugs in Ophthalmology.

This review focuses on 5 new anti-VEGF drugs in the advanced stage of clinical development (i.e., phase 3): conbercept, brolucizumab, port delivery system with ranibizumab, abicipar pegol and faricimab. Results of clinical trials and the advantages of each drug compared to the available molecules are discussed in detail.

Comparison of the Efficacy of Dexamethasone, Nepafenac, and Bromfenac for Preventing Pseudophakic Cystoid Macular Edema: an Open-label, Prospective, Randomized Controlled Trial.

PURPOSE: To evaluate the incidence of pseudophakic cystoid macular edema (PCMO) in patients treated with corticosteroids alone or in combination with bromfenac or nepafenac eyedrops after uneventful cataract surgery. MATERIALS AND METHODS: Prospective, randomized, open-label, placebo-controlled clinical trial. Patients who underwent routine cataract surgery with intraocular lens (IOL) implant were randomly divided into three groups receiving either bromfenac or nepafenac in association with dexamethasone or dexamethasone alone (control group) postoperatively. Best-corrected visual acuity (BCVA) measurement, slit lamp and fundus examination and optical coherence tomography (OCT) were performed preoperatively, 1 and 5 weeks after surgery. Primary outcome was defined as patients (%) in whom macular edema developed within 5 weeks after cataract surgery; secondary end points were patients (%) with BCVA improvement from pre-op through 5 weeks after surgery and corneal toxicity. RESULTS: A total of 144 patients completed the study, 48 for each group. In all groups, mean central subfield thickness at OCT increased significantly 5 weeks after surgery (p < 0.01). However, at this time point, four patients (8.3%) of the control group and none in nepafenac and bromfenac groups developed PCMO (p = 0.016). Compared with baseline, mean BCVA significantly improved both at 1 and 5 weeks in all groups (p < 0.01). At 1 week, the nepafenac group showed a mean BCVA significantly lower compared with both the control (p = 0.038) and bromfenac group (p = 0.002). CONCLUSIONS: Co-administration of nepafenac or bromfenac and steroids in patients who underwent routine cataract surgery is associated with a lower incidence of PCMO compared with steroid monotherapy.

Homozygous Recessive Versican Missense Variation Is Associated With Early Teeth Loss in a Pakistani Family.

Only a few genes involved in teeth development and morphology are known to be responsible for tooth abnormalities in Mendelian-inherited diseases. We studied an inbred family of Pakistani origin in which two first-cousin born brothers are affected by early tooth loss with peculiar teeth abnormalities characterized by the absence of cementum formation. Whole exome sequencing revealed a H2665L homozygous sequence variant in the VCAN gene. Dominant splicing mutations in VCAN are known to cause Wagner syndrome or vitreoretinopathy. We explored teeth morphology in these two patients, while versican expression was assessed by western blot analysis. Early signs of vitreoretinopathy were found in the elder brother while the parents were completely negative. Our findings suggest that the homozygous recessive H2665L missense sequence variant impairs the normal morphology of the teeth roots via loss of cementum synthesis, and is also associated with early onset, recessive, Wagner syndrome, thus expanding both the phenotype mutation scenario and the inheritance mode of VCAN mutations.

Anti-VEGF Therapy for Retinal Vein Occlusions.

Retinal vein occlusion (RVO) is the second most common cause of visual loss in the Western World. RVO is usually classified into branch RVO (BRVO) and central RVO (CRVO) according to the anatomical site of the vascular occlusion. The pathogenesis of RVO is not yet fully understood, however an important event is the intraluminal thrombus formation, which is usually secondary to several conditions such as hypertension, hyperlipidemia, diabetes and thrombophilia. The blockage of venous circulation causes an elevation of intraluminal pressure in the capillaries, leading to hemorrhages and leakage of fluid within the retina, increase of interstitial pressure and a consequent reduction of retinal perfusion. Ischemia may develop resulting in secretion of vascular endothelial growth factor (VEGF) that causes further vascular leakage and retinal oedema. VEGF has therefore a leading role in RVO pathogenesis and symptoms. As a consequence use of anti-VEGF agents by intravitreal injections has become very common with the aim to improve the clinical outcomes in these patients. Currently 2 anti-VEGF agents (ranimizumab and aflibercept) have been FDA (Food and Drug Administration) and EMA (European Medicine Agency) approved for the treatment of RVO, while another VEGF inhibitor (bevacizumab) is often used "off-label" in clinical practice. Many treatment regimens have been suggested in the clinical trials with these drugs, as monthly injections or injections when needed, however the ideal regimen has not been defined yet. We conducted a systematic review searching MEDLINE for the following terms: retinal vein occlusion, ranibizumab, bevacizumab, aflibercept, vascular endothelial growth factor, macular oedema. Data were extracted by one author (AG and BE) and checked by a second (BPM, CC). Aim of this article was to review available data for each drug, focusing on their efficacy and safety trying to compare their advantages and limits.

Fundus autofluorescence patterns in Best vitelliform macular dystrophy.

PURPOSE: To provide a systematic classification of fundus autofluorescence (FAF) patterns in patients affected by Best vitelliform macular dystrophy. DESIGN: Cross-sectional prospective study. METHODS: Patients affected by Best vitelliform macular dystrophy at different stages of the disease were prospectively enrolled from January 2012 to July 2013. Eighty eyes of 40 patients were included in the study. All patients underwent a complete ophthalmologic examination, including genetic characterization, short-wavelength FAF, and near-infrared FAF. Main outcome measures were the recognition of the FAF patterns in the different stages and the identification of a relationship between FAF patterns and best-corrected visual acuity (BCVA). RESULTS: Six FAF patterns for both short-wavelength and near-infrared FAF were identified, including normal, hyper-autofluorescent, hypo-autofluorescent, patchy, multifocal, and spoke-like patterns. Applying Gass's classification for defining consecutive stages of Best vitelliform macular dystrophy (namely vitelliform, pseudohypopyon, vitelliruptive, atrophic, and cicatricial) identified no pattern as stage-specific. Patchy patterns had the highest prevalence. A statistically significant difference (Kruskal-Wallis ANOVA) was found among hyper-autofluorescent, patchy, and hypo-autofluorescent patterns, both in short-wavelength (P = .001) and near-infrared FAF (P = .001). Hyper-autofluorescent and hypo-autofluorescent patterns were associated with better and worse BCVA, respectively. CONCLUSIONS: Six main patterns on both short-wavelength and near-infrared FAF were identified in Best vitelliform macular dystrophy. No FAF pattern can be considered stage-specific. Although a difference in the BCVA among the FAF patterns was registered, only a longitudinal study designed to evaluate the clinical and FAF modifications over the follow-up will help clarify the prognostic implications of each FAF pattern.

Effect of VEGF and anti-VEGF compounds on retinal pigment epithelium permeability: an in vitro study.

OBJECTIVE: To evaluate the effect of 2 vascular endothelial growth factor (VEGF) isoforms (121 and 165) and 2 anti-VEGF compounds (ranibizumab and pegaptanib sodium) on the permeability of human retinal pigment epithelium (RPE) cells in vitro.
 METHODS: The RPE permeability was assessed on ARPE19 cells grown onto inserts of polytetrafluoroethylene previously treated with ammonia gas plasma. Paracellular permeability to ions was measured by mean of transepithelial electrical resistance (TEER). Permeability to non-ionic molecules was gathered by the amount of fluorescein dextran (FD) passing across the monolayer within 2 hours.
 RESULTS: Only VEGF165 applied at the apical side of the monolayer induced a statistically significant decrease of TEER (p<0.001). No changes in TEER were observed when pegaptanib sodium or ranibizumab were apically administered together with VEGF165. 
Both VEGF isoforms significantly increased permeability to 4 kDa dextran (p<0.01). Apical administration of ranibizumab or pegaptanib sodium as well as coadministration of pegaptanib sodium with VEGF121 or VEGF165 induced a statistically significant increase of permeability to 4 kDa FD. 
 CONCLUSION: Both VEGF isoforms and anti-VEGF compounds exert an effect on human RPE permeability in vitro.

Computerized assessment of intraretinal and subretinal fluid regions in spectral-domain optical coherence tomography images of the retina.

PURPOSE: To evaluate a new computerized segmentation technique for the quantification of intraretinal and subretinal fluid in spectral-domain optical coherence tomography (SD OCT) images of the retina. DESIGN: Prospective, cross-sectional study. METHODS: Thirty-seven B-scan images of 37 patients with exudative age-related macular degeneration were chosen randomly from SD OCT volume scans (1 per volume scan). All hyporeflective areas in the image first were segmented automatically as candidate regions by the program. Researchers who were masked to the candidate region information selected each fluid region from the original image using a single mouse click. The program then delineated the boundary of each region selected and calculated quantitative parameters, including total area of fluid regions if multiple regions were selected. The performance of our technique was validated by comparing the results with the measurements obtained from boundaries manually delineated by 2 masked observers. Time efficiency, agreement with manual delineation, and intraobserver and interobserver agreement of using the program were evaluated. RESULTS: The proposed technique reduced the average processing time per image approximately 6-fold (15 seconds for computerized segmentation vs 90 seconds for manual delineation). There was good agreement between computerized segmentation and manual delineation measured by intraclass correlation coefficient (range, 0.897 to 0.979) and the Dice coefficient (range, 0.721 to 0.785). The proposed technique has excellent intraobserver and interobserver agreement (intraclass correlation coefficient range, 0.998 to 0.999; Dice coefficient range. 0.959 to 0.981). CONCLUSIONS: This computerized segmentation method allows for accurate and fast quantification of fluid in retinal SD OCT images and could assist in monitoring disease progression and evaluating therapeutic intervention.

Early multifocal electroretinogram findings during intravitreal ranibizumab treatment for neovascular age-related macular degeneration.

PURPOSE: To evaluate changes in the multifocal electroretinogram (mfERG) in patients with neovascular age-related macular degeneration (nAMD) undergoing ranibizumab treatment. METHODS: This was an observational, longitudinal, prospective study. Treatment-naive patients with nAMD who met the inclusion and exclusion criteria underwent a course of monthly injections of ranibizumab over 3 months. At baseline and month 3, each subject was evaluated with best corrected visual acuity (BCVA), contrast sensitivity (CS), fluorescein and indocyanine green angiography, optical coherence tomography (OCT), and mfERG. Additional mfERGs were performed at weeks 1 and 4 and BCVA and OCT at weeks 4 and 8. RESULTS: Eighteen patients were enrolled. Between baseline and week 12, median BCVA improved from 59 to 69 ETDRS letters (P = 0.001), median CS improved from 29 to 30 letters (P = 0.05), mean OCT central foveal subfield thickness (CFT) decreased from 294 to 199 µm (P = 0.005), mean P1 amplitude density of the mfERG central zone increased from 35.85 to 51.55 nV/deg(2) (P = 0.009). The mfERG response correlated positively with BCVA (F = 22; P < 0.0001) and negatively with CFT (F = 12.73; P = 0.00078). CONCLUSIONS: Intravitreal ranibizumab therapy appears to induce an increase in mfERGs centrally in patients with nAMD at least in the short term. Longer term studies to investigate the prognostic value of mfERG responses to predict changes in visual acuity in nAMD and other diseases are warranted. (ClinicalTrials.gov number, NCT01023971.).

Intraocular pressure changes following the use of silicone oil or Densiron 68 as endotamponade in pars plana vitrectomy.

OBJECTIVE: To compare the effects of standard silicone oil 5700 (SSO) and heavy silicone oil (HSO) such as Densiron(®) 68 on intraocular pressure (IOP). MATERIALS AND METHODS: Retrospective case series including 180 eyes (105 treated with SSO and 75 with HSO). IOP was measured before surgery, 1 day after, and then at 1-, 3-, 6-, and 12-month follow-ups. RESULTS: In the SSO group, a significant increase in IOP occurred in 14% of the eyes (15/105) at 1 day postoperatively, and persisted in 11.4% (12/105) at 1-month follow-up. In the HSO group, a persistent elevated IOP was recorded in 20% of the eyes (15/75) at 1 day postoperatively, and in 16% (12/75) at 1-month follow-up. At 12-month follow-up, mean IOP was 16.7 ± 8.7 mmHg and 19.7 ± 3.8 mmHg, respectively, in the SSO and HSO groups. The difference between the 2 groups was always not significant. CONCLUSION: Overall, the use of Densiron 68 was not associated with higher IOP values as compared with SSO.

Inflammatory mediators and angiogenic factors in choroidal neovascularization: pathogenetic interactions and therapeutic implications.

Choroidal neovascularization (CNV) is a common and severe complication in heterogeneous diseases affecting the posterior segment of the eye, the most frequent being represented by age-related macular degeneration. Although the term may suggest just a vascular pathological condition, CNV is more properly definable as an aberrant tissue invasion of endothelial and inflammatory cells, in which both angiogenesis and inflammation are involved. Experimental and clinical evidences show that vascular endothelial growth factor is a key signal in promoting angiogenesis. However, many other molecules, distinctive of the inflammatory response, act as neovascular activators in CNV. These include fibroblast growth factor, transforming growth factor, tumor necrosis factor, interleukins, and complement. This paper reviews the role of inflammatory mediators and angiogenic factors in the development of CNV, proposing pathogenetic assumptions of mutual interaction. As an extension of this concept, new therapeutic approaches geared to have an effect on both the vascular and the extravascular components of CNV are discussed.

Automated segmentation of foveal avascular zone in fundus fluorescein angiography.

PURPOSE. To describe and evaluate the performance of a computerized automated segmentation technique for use in quantification of the foveal avascular zone (FAZ). METHODS. A computerized technique for automated segmentation of the FAZ using images from fundus fluorescein angiography (FFA) was applied to 26 transit-phase images obtained from patients with various grades of diabetic retinopathy. The area containing the FAZ zone was first extracted from the original image and smoothed by a Gaussian kernel (sigma = 1.5). An initializing contour was manually placed inside the FAZ of the smoothed image and iteratively moved by the segmentation program toward the FAZ boundary. Five tests with different initializing curves were run on each of 26 images to assess reproducibility. The accuracy of the program was also validated by comparing results obtained by the program with the FAZ boundaries manually delineated by medical retina specialists. Interobserver performance was then evaluated by comparing delineations from two of the experts. RESULTS. One-way analysis of variance indicated that the disparities between different tests were not statistically significant, signifying excellent reproducibility for the computer program. There was a statistically significant linear correlation between the results obtained by automation and manual delineations by experts. CONCLUSIONS. This automated segmentation program can produce highly reproducible results that are comparable to those made by clinical experts. It has the potential to assist in the detection and management of foveal ischemia and to be integrated into automated grading systems.

Topical and oral ketorolac administration increases the intraocular pressure-lowering effect of latanoprost.

PURPOSE: To verify the influence of a non-steroidal anti-inflammatory drug (NSAID), ketorolac (topical and oral) on the intraocular pressure reduction induced by 0.005% latanoprost topical administration, both in patients affected by primary open-angle glaucoma and in healthy controls. METHODS: Two groups of subjects were enrolled for this randomized, prospective, masked clinical study: 16 glaucomatous patients well controlled with 0.005% latanoprost eyedrops (group I) and 16 healthy adult volunteers (group II). Group I subjects were treated at one-week intervals with 10 mg of oral ketorolac, oral placebo, topical ketorolac, and topical placebo, respectively; for each administration modality, the switch between drug and placebo was performed in a randomized, crossover, double-blind fashion. Group II subjects followed the same protocol, with the topical once-daily 0.005% latanoprost treatment starting three days prior to the ketorolac/placebo administration. Intraocular pressure (IOP) was investigated in both groups on the day of oral/topical administration of ketorolac or placebo at baseline (8:00 AM) and at the following intervals: 1, 2, 4, 8, 12, and 24 hours. RESULTS: No significant IOP changes after oral and topical placebo administration were observed in either group. In contrast, when the subjects received ketorolac (either oral or topical), a marked decrease in IOP was recorded, with a noticeable fall at the first hour after the NSAID administration (p = 0.01), which remained still significant 8 hours later (p < 0.05). CONCLUSION: Topical and oral ketorolac strengthens the latanoprost-induced IOP-lowering effect both in glaucomatous patients and in healthy subjects.