bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proteomic analysis of aqueous humor in retinoblastoma: Final results
Authors: MICHELI L , LANDI G , MORIGGI M , VANNONI D , VASSO M , MENICACCI F , GELFI C , LEONCINI R , HADJISTILIANOU T 
Published: November 2013

AbstractPurpose Proteomics in ocular fluids is gaining rapidly relevance.Since 2009 we work on the analysis of proteic composition of the aqueous humor (AH) from retinoblastoma (RTB) patients and,in this work,we use the 2D‐DIGE coupled to mass spectrometer to find new RTB markers.Methods We enrolled 22 RTB patients(Reese‐Ellsworth stage V or ABC classification group E)undergoing enucleation and 20 normal subjects undergoing cataract surgery(CTR).Five of 22 patients presented with associated secondary glaucoma whereas 17 had no secondary glaucoma;8 of 17 patients with no secondary glaucoma received chemotherapeutical treatment with melphalan.Pools were analyzed using 2D‐DIGE coupled to mass spectrometer.DIA and BVA were fully automated with 2D DeCyder® and statistical analysis was performed with EDA® module.Significant spots were In‐Gel digested and identified with Maldi‐TOF/TOF.Results PCA analysis of the data underlines high distance between CTR pool and RTB pools.In particular the analysis of single protein shows that all the RTB pools are characterized by a high level of Crystallin and pro‐inflammatory proteins and CTR pool are characterized by a high level of Epithelium Growth Factor.Conclusion This study is the last step for the clarification of AH protein pattern in patients with RTB.Although we learn from literature that the presence of inflammatory processes is widely documented in late stages of pathology,however the presence of crystalline proteins could be either a new marker or an active part of the late stage tumor‐induced panorama in RTB AH samples.Due to this,we think that several molecular biology studies are necessary to better understand involvement of this protein family in RTB pathological pathway.
Pigmented basal cell carcinoma of the eyelid
Authors: ROSNER M , PRIEL A , BEN SIMON1 G , ROSEN N 
Published: November 2013

AbstractPurpose While Basal cell carcinoma (BCC) is the most common malignant lesion of the eyelid, pigmented basal cell carcinoma of the eyelid are only rarely seen. Thus it is usually misdiagnosed clinically. The purpose of this presentation is to describe three cases with pigmented periocular BCC and to discuss its clinical and histopathological characteristics.Methods A retrospective review was performed, of the files and the histopathologic slides of patients with pigmented BCC of the eyelid, who were treated in the Goldschleger Eye Institute, Sheba Medical Center, Tel Hashomer, Israel, during 2007‐2012.Results Fifty three patients were found with eyelid BCC diagnosed between 2007 and 2012, and three of them had pigmented BCC. Two of the tumors were on the lower lid and one in the medial canthus. All were uniformly or almost uniformly darkly pigmented. All were nodular and treated by complete surgical excision. The clinical diagnosis of two of the lesions was nevus.Conclusion Pigmented BCC should be remembered among the differential diagnosis of pigmented lesions of the eyelid. It is interesting that our cases of pigmented BCC such as other cases described in the literature, were nodular or nodulo‐ulcerative and not cystic or sclerosing types.
Pro‐inflammatory cytokines induce apoptosis of human retinal capillary endothelial cells through downregulation of Hsp27
Authors: NAGARAJ R , NAHOMI R 
Published: November 2013

AbstractPurpose Retinal capillary cells undergo apoptosis in diabetes, but the mechanism is not clear. Pro‐inflammatory cytokines are upregulated in the diabetic retina of humans and rodents. In this study, we have investigated the effect if pro‐inflammatory cytokines on a small heat shock protein, Hsp27 in human retinal endothelial cells (HREC)Methods HREC were cultured in the presence of pro‐inflammatory cytokines, interferon ‐γ (IFN‐γ, 50 and 100 units/ml), interleukin‐1β (IL‐1β, 10 and 20 ng/ml) and tumor necrosis factor‐α (TNF‐α, 10 and 20 ng/ml) for 48 hrs and in the presence or absence of high glucose (25 mM, HG). The roles of the kynurenine pathway and NOS were determined by adding 20 μM 1‐methyl tryptophan (MT) and 500 μM L‐Nω‐nitroarginine methyl ester hydrochloride (L‐NAME), respectively to the culture medium.Results HREC cultured in the presence of mixed cytokines showed a significant downregulation of Hsp27 at the protein and mRNA levels. This downregulation was not due to downregulation of the transcription factor, HSF‐1. Mixed cytokines activated indoleamine 2,3‐dioxygenase (IDO), enhanced kynurenine production and increased the ROS content in HREC. MT inhibited the effects of mixed cytokines on Hsp27. Mixed cytokines upregulated NOS2 and increased levels of NO. A peroxynitrite donor and exogenous peroxynitrite drastically reduced Hsp27. The cytokine and HG‐mediated downregulation of Hsp27 was accompanied by increased apoptosis of HRECConclusion Our data suggest that pro‐inflammatory cytokines induce ROS and NO formation, which through peroxynitrite production reduce Hsp27 and bring about apoptosis of HREC. These results suggest a novel mechanism for capillary cell death in diabetic retinopathy.
Characterization of rafts subtypes in monocytes of diabetic patients with and without neovascular complication
Authors: MATHIEU B , GAMBERT‐NICOT S , BRETILLON L , BRON AM , CREUZOT‐GARCHER C 
Published: November 2013

AbstractPurpose Diabetic retinopathy is a chronic inflammatory disease where the infiltration of plasma and neovascularization of the retina cause blindness. This disease affects millions of people in developed countries and early biological markers would better manage these patients.Methods For this purpose we purified monocytes, key players of inflammation, blood of healthy and sick people. On the surface of monocytes, there are dynamic structures called rafts membrane microdomains, which there are two subtypes: flat rafts and caveolae rafts. They correspond to dynamic assemblies and ordered cholesterol and sphingolipids. We isolated these rafts membranes of monocytes and studied protein AT2 (angiotensin receptor II) and gp91 (major membrane subunit of NADPH oxidase, producing enzyme superoxide anions), potential markers of the severity of diabetic retinopathy.Results Our results have shown a switch of cholesterol and sphingomyelin flat rafts to caveolae rafts. The analysis of the constituent proteins (flotillin and caveolin) and functional (AT2, gp91 and p47phox) have shown that these are mobile in the membrane in an inflammatory context. Subunits of NADPH oxidase seem to gather in caveolae rafts and more precisely when the enzyme is activated.Conclusion The proteins of interest and the switch composition of lipid rafts therefore would appear to be an indicator of the progression of diabetic retinopathy and it would be interesting to provide a mechanistic explanation for the evolution of this chronic vascular disease to consider potential therapeutic targets.
The UK Sight Loss and Vision Priority Setting Partnership (SLV‐PSP): Vision research questions prioritised by patients and health care professionals
Authors: VOTRUBA M , THE SLV‐PSP STEERING GROUP AT 
Published: November 2013

AbstractPurpose The SLV‐PSP process aims to identify research priorities that can be used to inform decisions of funding bodies and enhance the case for additional research funding.Methods People were asked to submit questions about prevention, diagnosis and treatment of sight loss and eye conditions that are most important to them. The survey was available online, by phone, post and alternative formats (Braille and audio). The partner organisations were Fight for Sight, the College of Optometrists, NIHR Biomedical Research Centre for Ophthalmology, The Royal College of Ophthalmologists and Vision 2020. Over 30 charitable and professional organisations promoted the survey.Results 2,200 people responded generating 4,461 questions. 65% of respondents had sight loss or an eye condition. 17% were healthcare professionals (ophthalmologists, optometrists, orthoptists, ophthalmic nurses, opticians, social care/ rehabilitation). The questions were allocated to 12 eye disease categories before subsequent prioritisation of the top 10 at 12 workshops (questions at http://fightforsight.org.uk/sightlosspsp). The categories were ARMD, cataract, corneal and external, glaucoma, inherited retinal, neuro‐ophthalmology, ocular cancer, ocular inflammatory, refractive error and ocular motility and vitreoretinal/ocular trauma.Conclusion Analysis of demographic data for respondents indicates that the sample is generally representative of the UK population. The number and diversity of conditions represented in responses is distributed across the major conditions and fits projected population incidence.
Visual acuity thresholds associated with activity limitations in the elderly. The POLA study
Authors: DAIEN V , PERES K , VILLAIN M , COLVEZ A , CARRIERE I , DELCOURT C 
Published: November 2013

AbstractPurpose Activity limitations, which induce loss of autonomy in the elderly, are a major public health problem. We investigated the associations between objectively determined visual impairments and activity limitations, and assessed the visual acuity thresholds associated with these restrictions.Methods The study sample consisted of 1887 people aged 63 years and over from a population‐based cohort. Visual impairment was defined according to the WHO definition. Multivariate logistic regressions were used to estimate the associations between vision and Instrumental Activities of Daily Living (IADL) limitations. Using receiver‐operating characteristic (ROC) curves, we identified visual acuity thresholds that maximized the Youden index ([Sensitivity+Specificity]‐1) for predicting IADL limitations.Results After adjustment for potential confounders, moderate to severe visual impairment and mild visual impairment were strongly associated with IADL limitations(odds ratio [OR]=3.49; 95% confidence interval [CI]=1.93, 6.32 and OR=1.77; 95% CI=1.07, 2.91, respectively). Visual acuity was a strong predictor of IADL limitations, with an area under the ROC curve of 0.72 (95% CI=0.68, 0.76). The best discrimination between subjects with or without IADL limitations (global, physical, cognitive) was obtained for visual acuities around 20/40‐20/50.Conclusion This study confirms major increased risk for IADL limitations in subjects with moderate to severe visual impairment. In addition, it suggests that milder visual impairments (in particular below 20/40) may also be related to an increased risk for IADL limitations and should be considered for early medical intervention, before the decline of the subjects’ autonomy.
Functional characterization of retinitis pigmentosa causing RPGR mutations
Authors: SHU X , PATNAIK S , CRAFT J , MCCULLOCH D 
Published: November 2013

AbstractPurpose Retinitis pigmentosa (RP) is a progressive outer retinal dystrophy, affecting 1/3500 individuals in most populations. X‐linked RP (XLRP) is one of the most severe forms of human retinal degeneration, as determined by age‐of‐onset and progression. Mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene, responsible for the major subtype of XLRP, are the most common single cause of RP, accounting for 6‐20% of all cases. We aim to investigate the functional role of RPGR mutations in the pathogenesis of XLRP.Methods Different RP‐causing RPGR mutations were cloned into pCEP‐4 expression vector and transfected into retinal pigment epithelium (RPE) 1 and IMCD3 cell lines. RPGR expression was detected by immunocytochemistry and western blotting. The pathological effects caused by RPGR mutations were characterized by immnocytochemistry.Results The average length of cilia in RPE1 cells with over‐expression of mutant RPGR are around 50‐60% shorter compared to that in cells with over‐expression of wild type RPGR. Cells with over‐expression of RPGR mutant proteins exhibited stronger actin filaments. The frameshift RPGR mutations also cause aggregations in IMCD3 cell lines indicating that the mutant proteins are misfolded which may cause toxicity.Conclusion Mutant RPGR proteins have functional role in cilia defect through regulating actin remodelling,
Modern Trabeculectomy: Technique, and results from a UK muticentre study
Authors: KIRWAN JF , LOCKWOOD A , MCNAUGHT AI , MACLEOD A , SHAH P , KING AJ , BROADWAY DC , AGRAWAL P 
Published: November 2013

AbstractEfficacy and safety of current trabeculectomy surgery Methods: 428 patients with OAG and no previous glaucoma surgery from 9 units were evaluated. All cases had 2 yr minimum follow up. Main outcomes: IOP, VA, complications and interventions. Reoperation for glaucoma (except needling and resuturing) and loss of light perception were classified as failures. Results: Antifibrotics were used in 400 (93%) cases; MMC in 271 (63%), 129 had 5‐FU (30%). At 2 years, IOP (mean ± SD) was 12.3 ± 4.3 mmHg, 370 (86%) had IOP ≤21 mmHg without medication; 405 (95%) achieved an IOP ≤ 21 mmHg overall. IOP ≤18 mm Hg was achieved by 352 (82%) without treatment and 373 (87%) overall. By TVT criteria: 310/365 (85%) had complete success at 2 yr and 335/365 (92%) achieved qualified success. Postoperative treatments: suture manipulation 43%, resuturing 5.1%, bleb needling 16%, postop 5FU 27% and cataract extraction in 31%. Visual loss of more than two lines occurred in 24 (5.6%). 31 (7.2%) had late hypotony (IOP <6 mmHg). 2 patients (0.5%) developed blebitis. One patient had endophthalmitis at 3 wks (0.2%). We show good trabeculectomy outcomes with low rates of complications can be achieved but intensive postoperative care is required.
In vivo integrity of intra‐corneal bioengineered discs in rabbit models
Authors: RAFAT M , LAGALI N , KOULIKOVSKA M , GRIFFITH M , FAGERHOLM P 
Published: November 2013

AbstractBackground: We have previously reported the successful integration and safety of bioengineered materials as corneal substitutes in human models. Despite the promising results as corneal implants, more elastic and robust materials are required for use as thin intra‐corneal lenses to withstand surgical manipulation for corrective surgery and improved vision. Most of the existing corneal inlays are made of synthetic materials. Here we describe the potential of bioengineerd materials for vision correction. Objectives: to develop bioengineered materials as inlays within the corneal tissue as well as evaluating the in vivo integrity and integration of the materials in rabbit models. Methods: Bioengineered inlays were prepared from collagen and tested for their physical and biological propertis. A femtosecond laser was used to cut 100 mircon thick discs of mid‐stromal tissue from corneas of 20 rabbits and replaced with bioengineered inlays. Results: The new materials demonstrated improved mechanical properties while maintaining their clarity and biocompatibility. The bioengineered inlays retained their shapes, thickness, and clarity 8 weeks post‐surgery in rabbits.
Commercial interest
The basics of femtosecond laser cataract surgery
Authors: NAGY Z 
Published: November 2013

AbstractFemtosecond lasers have been used for corneal flaps for years. Recently the problems of beam delivery through the ocular optic media has been solved. Precise incisions can be performed within the crystalline lens (fragmentation or liquefaction), on the anterior capsule and possible on the posterior capsule as well (capsulotomy) and in the cornea (customised geometry, size and position of the wounds). During the lecture the basics of femtolaser catarct surgery will be shown.
Endothelial graft precutting from the epithelial and endothelial side with the femtosecond laser, on cornea stored in a new corneal bioreactor
Authors: BERNARD A , MAUCLAIR C , BAUBEAU E , GRANIER J , EGAUD G , HE Z , CAMPOLMI N , PISELLI S , FOREST F , PEOC'H M , DUMOLLARD JM , THURET G , GAIN P 
Published: November 2013

AbstractPurpose To compare femtosecond laser endothelial graft precut from epithelial and endothelial side, on corneas stored in a new corneal bioreactorMethods Human corneas unsuitable for transplantation for serological reasons and procured by the Auvergne‐Loire eye bank (Saint Etienne, France) were used after informed consent of the relatives, as authorized by French bioethics laws. Femtosecond laser lamellar cuts were realized on corneas stored in a new corneal bioreactor, patented by our laboratory, designed to maintain a constant endothelial pressure and to permit an epithelial aplanation without opening the device. Lamellar cuts of several thickness were realized with corneal epithelium or corneal endothelium side up, but always with epithelial aplanation. OCT images (Casia SS1000, Tome) were realized in the bioreactor before and after each cut. Endothelial viability was determined by using a triple Hoechst/Ethidium/Calcein labeling (HEC, IOVS Pipparelli 2011) coupled with image analysis of the whole endothelial surface immediately after the cut. Surface roughness was assessed by scanning electron microscopyResults This study demonstrates the possibility to realise femtoseconde laser endothelial graft precut directly in a dedicated storage device. First results of endothelial viability in function of lamellar graft thickness and lamellar cut side will be presented as well as surface roughnessConclusion Capacity to realize femtosecond laser endothelial graft precut directly in a storage device could improve the quality and security of delivered lamellar grafts
Adeno‐associated viral vector serotypes for gene transfer to corneal endothelial cells
Authors: FUCHSLUGER T , MUELLER C , DANA R 
Published: November 2013

AbstractPurpose The aim of this study was to determine the efficacy and efficiency of the non‐pathogenic recombinant adeno‐associated viral vectors (rAAV) for gene transfer to EC. Therefore, different rAAV pseudotypes were compared to assess the expression and kinetic patterns of a reporter gene (GFP).Methods A comparison of GFP expression and kinetics after EC transduction using a AAV 2/1, 2/2, 2/5, 2/8, 2/9, 2/10 was performed on both murine EC (Balb/C) and human EC (corneas, cell line and primary cells). In addition, the effects of different vector concentrations (3x10^3/10^4, 10^5, 10^6, 10^7, 10^8IU/µl) were investigated. Analyses were performed using laser scanning microscopy and flow cytometry.Results Significant differences between human and murine EC as well as primary and immortalized EC were detected. Murine corneal EC transduced with AAV2/2, 2/5 and 2/9 resulted in considerable protein expression, AAV 2/5 did not show considerable expression in human corneas. A slow onset of GFP expression both in immortalized and primary EC was detected, leading to peak and stable expression around 2‐3 weeks. Peak expression was reached earlier in primary EC and immortalized murine EC compared to immortalized human EC. The overall plateau of expression was highest in primary EC (~80%).Conclusion Pseudotypes of recombinant AAV alter in their tropism and transduction efficiency between murine and human corneas. In general high levels of gene expression can be obtained with several of these pseudotypes. Characteristic slow‐onset kinetics of protein expression have to be taken into account when applying AAVs in translational applications, e.g. in corneal storage in eye banks.
Iron antagonism of DICER1 promotes NLRP3 inflammasome priming due to enhanced Alu RNA stability
Authors: GELFAND B , KIM Y , YASUMA T , LI S , FOWLER B , KLEINMAN M , AMABTI J 
Published: November 2013

AbstractPurpose Excessive free iron induces retinal toxicity in several human diseases and has been linked to age‐related macular degeneration. Iron toxicity is widely attributed to its ability to catalyze hydroxyl radical formation through Fenton’s reaction. Iron was recently described as an inhibitor of canonical DICER1 enzymatic activity through sequestration of the cofactor poly(C)‐binding protein 2 (PCBP2). We sought to determine whether iron similarly impaired non‐canonical DICER1 activity to clear cytotoxic Alu RNA in the retinal pigemented epithelium.Methods Iron overload was induced in human ARPE‐19 cells by supplementing culture media with ferric ammonium citrate for 72 hours and in wild‐type mice by subretinal injection. Alu, B1 and B2 RNA abundance was measured by northern blotting. Alu RNA/DICER1/PCBP2 binding was assessed by immunoprecipitation, western blotting and LC‐MS. Alu RNA cleavage efficiency was evaluated using synthetic in vitro transcribed Alu RNA and recombinant human DICER1 and PCBP2. Inflammasome priming was assessed by quantifying NLRP3 mRNA via qRT‐PCR.Results Iron overload in cells and mouse retina induces robust accumulation of Alu transcripts as well as the rodent homologs B1 and B2 RNAs. Iron overload impairs Alu RNA clearance by RPE cells, independently of DICER1 mRNA abundance. Alu RNA binds to the iron sensitive co‐factor PCBP2, which enhances DICER1‐mediated Alu RNA cleavage. Iron overload or siRNAs targeting PCBP2 prime the NRLP3 inflammasome, which can be prevented by antisense‐mediated Alu RNA antagonism.Conclusion These data suggest that Alu RNA induced inflammasome signaling could contribute to retinal iron toxicity in addition catalytic free radical generation.
Retinal pigment epithelium cell‐derived microparticles mediate oxidative stress‐induced retinal cells dysfunction
Authors: TAHIRI H , YANG C , DUHAMEL F , CHEMTOB S , HARDY P 
Published: November 2013

AbstractPurpose Age‐related macular degeneration (AMD) represents the leading cause of vision loss in the elderly. The cumulative oxidative injury induces retinal pigment epithelium (RPE) membrane microparticles production, RPE cell death and cellular senescence. The RPE blebs are implicated in the formation of sub‐retinal deposit. Nonetheless, the pathophysiological roles of RPE microparticles (RMPs) remain largely unexplored. This study was designed to investigate whether RMPs participate in the retinal cells dysfunction.Methods RMPs and fluorescent DiI‐labelled RMPs were isolated from cultured ARPE‐19 cells under oxidative stress. RMPs‐treated RPE cells were subjected to WST‐1, cellular senescent, apoptotic assay and FACS cell cycle analysis respectively. The antibody against CD36 was used in uptake experiment to determine the involvement of scavenger receptor CD36.Results Our study revealed that uptake of RMPs by RPE cells is time‐dependent, and this process is partially dependent on CD36 evidenced by an approximately 50% decrease of RMPs uptake caused by CD36 antibody treatment. In addition, RMPs significantly reduced RPE cell viability in a dose‐dependent manner. RMPs in a concentration of 5% µg/ml significantly induced RPE cell‐cycle arrest at G0/G1 phase. RMPs‐treated cells exhibited a 19% increase in G0/G1 phase, with associated increases of the senescence‐associated β‐galactosidase activity.Conclusion We demonstrated for the first time that RPE cells uptake microparticles derived from RPE cells under oxidative stress. These findings strongly suggest that RMPs function as mediators to exacerbate the oxidative damages to RPE cells, and indicate a pathological role of RMPs in AMD.