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Background/Aims: We performed co-culture experiments between human RPE cells (ARPE- 19) and human umbilical vascular endothelial cells (HUVEC) in order to evaluate how anti- VEGF drugs could affect NO release, mitochondrial function, the oxidative status, proliferation and migration of RPE cells through modulation of their cross talk with vascular endothelial cells. Methods: The co-culture HUVEC/RPE, was exposed to Ranibizumab/Aflibercept in the absence/presence of the NO synthase (NOS) inhibitor, the phosphatidylinositol 3'-kinase (PI3K), the extracellular-signal-regulated kinases 1/2 (ERK1/2) and the p38 mitogen-activated protein kinase (p38 MAPK) blockers. Specific kits were used for cell viability, mitochondrial membrane potential, NO, ROS and GSH production. Western blot was performed for apoptosis markers, NOS isoforms, and others kinases detection. Cell migration was analyzed by scratch assay, whereas cell proliferation and cell cycle through xCELLigence and flow cytometry. Results: In RPE cells co-cultured with HUVEC in physiological conditions, Aflibercept/Ranibizumab increased NO release in a dose and time-dependent way. Opposite results were obtained in peroxidative conditions. Both anti-VEGF agents were able to prevent the fall of cell viability and mitochondrial membrane potential, an effect which was reduced by various inhibitors, and increased cell migration. Aflibercept/Ranibizumab counteracted the changes of apoptosis markers, NOS expression/activation, PI3K and ERK1/2 activation caused by peroxidation. These results were confirmed by cell cycle analysis. Conclusion: This study has shown new mechanisms at the basis of protective effects elicited by Aflibercept/Ranibizumab in RPE cells. HUVEC stimulated with Aflibercept/Ranibizumab, could release some paracrine factors that can modulate the RPE cells response in both physiologic and peroxidative conditions. [ABSTRACT FROM AUTHOR]

Purpose: Subthreshold micropulse laser (SMPL) has been increasingly used for the treatment of different retinal and choroidal macular disorders. However, the exact mechanisms of action have not yet been clearly defined. Therefore, we aimed to examine the role of SMPL treatment in the modulation of oxidant/antioxidant systems, apoptosis and autophagy in the mice eyes. Methods: A specific laser contact lens for retina was positioned on the cornea of 40 mice (20 young and 20 old) in order to focus the laser on the eye fundus for SMPL treatment. Within 6 months, 20 animals received one treatment only, whereas the others were treated three times. Eye specimens underwent histological analysis and were used for thiobarbituric acid reactive substances (TBARS) and glutathione (GSH) quantification, as well as for the superoxide dismutase 1 (SOD1) and the selenoprotein thioredoxin reductase 1 (TrxR1) expression evaluation. Western blot was performed for nitric oxide synthase (NOS) subtypes detection and to examine changes in apoptotic/autophagy proteins expression. Results: SMPL treatment reduced TBARS and increased GSH and SOD1 in the mice eyes. It also reduced cytochrome c, caspase 3 expression and activity and cleaved caspase 9, and increased Beclin 1, p62 and LC3β. The effects were more relevant in the elderly animals. Conclusion: Our results showed that SMPL therapy restored the oxidant/antioxidant balance within retinal layers and modulated programmed forms of cell death. Further studies may confirm these data and could evaluate their relevance in clinical practice. [ABSTRACT FROM AUTHOR]

Purpose: To investigate, in pediatric patients, the rate and clinical features of unspecific conjunctivitis of unknown origin (UCUO) and to evaluate their relationship with air pollution.Methods: From January to December 2013, we consecutively screened all of the patients referred for symptomatic ocular surface inflammation. Inclusion criteria for this study were age<14, diagnosis of conjunctivitis, residence in the Lombardia region. UCUO was defined as conjunctivitis of unknown etiology, not clearly due to infection or allergy. Based on addresses of residence and sites of 73 automatic air pollution monitoring stations (locations and mean annual particulate matter (PM) 10 and 2.5 concentrations were provided by the Regional Environmental Protection Agency (ARPA) Lombardia), each patient was paired with a value of exposure to PM. Relationship between UCUO and PM exposure was investigated.Results: A total of 132 of 251 screened children were included in this study. UCUO was diagnosed in 48/132 patients. The most common symptoms and signs in UCUO children were foreign body sensation (37/48) and conjunctival hyperemia (45/48), respectively. PM10 exposure value was significantly higher in UCUO (33.5±5.4µg/m3) compared to other groups (P<0.001, ANOVA). UCUO/total conjunctivitis ratio was significantly higher in residents in areas with more than 75 (Q3) days/year exceeding 50µg/m3 (EU legal limit) compared to areas with less than 45 (Q1) exceedances/year: 24/39: 61% vs 8/35: 23%; P>0.001, Chi-square test.Conclusion: Our data suggest a relationship between UCUO and air pollution. This form of conjunctivitis is not rare in pediatric patients and may be the most frequent in most polluted areas. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate corneal subbasal layer changes after uneventful cataract surgery by means of in vivo confocal microscopy. Methods: This prospective study included 30 patients. Before surgery, and 1, 3, 6, 8 and 10 months after cataract surgery, all patients underwent a complete ophthalmological and confocal microscopy examination in the central and temporal corneal areas. Number of fibers, beading, tortuosity and reflectivity were analyzed. Results: Important changes were shown in the central cornea up to 3 months after surgery: a reduction in nerve fiber number (baseline: 4.4 ± 1.7; month 1: 1.2 ± 0.5, p < 0.0001; month 3: 2.5 ± 1.2, p < 0.005) and reflectivity (baseline: 3.6 ± 0.5; month 1: 1.4 ± 0.6, p < 0.0001; month 3: 1.9 ± 0.9, p < 0. 0001), and an increase in beading (baseline: 0.3 ± 0.5 beads/100 µm2; month 1: 2.7 ± 0.6 beads/100 µm2, p < 0.0001; month 3: 2.6 ± 0.5 beads/100 µm2, p < 0. 0001). The confocal parameters completely progressively recovered thereafter (60% at 6 months, 87% at 8 and 10 months). The temporal plexus was absent at 1 month and fully recovered in all patients at month 8. Conclusion: Uneventful cataract surgery induces relevant corneal modifications when inspected by means of confocal microscopy. © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]