bluuebunny/crossref_metadata_2025_split huggingface dataset:
Celastrol regulates innate immunity response via NF‐kB and HSP70 in ARPE‐19 cells
Authors: PAIMELA T 
Published: November 2010

Abstract Purpose Chronic inflammation participates in the pathology of age‐related macular degeneration (AMD). Recent studies indicate that celastrol, a novel triterpene compound, modulates inflammatory responses, but its effect on the human retinal pigment epithelial cells is poorly understood. In this study, we investigated the potential anti‐inflammatory role of celastrol and its effect on nuclear factor kappa B (NF‐kB) activity in human retinal pigment epithelial cells (ARPE‐19). Methods ARPE‐19 cells were exposed to lipopolysaccharide (LPS; TLR 4 agonist) with simultaneous exposure to various concentrations of celastrol and the secretion of IL‐6 cytokine was analyzed by ELISA. The effect of celastrol exposure on heat shock protein 70 (HSP70) expression was analyzed by western blotting. In response to celastrol and modulated HSP70 levels NF‐kB activity was examined by ELISA. Results Celastrol suppressed the LPS‐induced IL‐6 expression levels via NF‐kB transcription factor in ARPE‐19 cells. Celastrol evoked elevated HSP70 levels without cytotoxicity. Interestingly, celastrols capability to inhibit NF‐kB activity was diminished when HSP70 response was suppressed by siRNA. This reveals that celastrol has potent anti‐inflammatory capacity in ARPE‐19 cells, and its effect is modulated through NF‐kB and HSP70. Conclusion Our findings reveal that celastrol is a novel compound to suppress innate immunity response in human retinal pigment epithelial cells.
p62/sequestosome 1 as a regulator of proteasome inhibitor‐induced autophagy in human retinal pigment epithelial cells
Authors: KAARNIRANTA K 
Published: November 2010

Abstract Purpose The pathogenesis of age‐related macular degeneration involves impaired protein degradation in retinal pigment epithelial (RPE) cells. The ubiquitin‐proteasome pathway and the lysosomal pathway including autophagy are the major proteolytic systems in eukaryotic cells. Prior to proteolysis, heat shock proteins (HSPs) attempt to refold stress –induced misfolded proteins and thus prevent the accumulation of cytoplasmic protein aggregates.The functional roles of p62 and HSP70 were evaluated in conjunction with protesome inhibitor ‐induced autophagy in human RPE cells (ARPE‐19). Methods The p62, HSP70 and ubiquitin protein levels and localization were analyzed by Western blotting and immunofluorescense. Confocal and transmission electron microscopy were used to detect cellular organelles and to evaluate the morphological changes. The p62 and HSP70 levels were modulated using RNA interference and overexpression techniques. Cell viability was measured by colorimetric assay. Results Proteasome inhibition evoked the accumulation of p62 and HSP70 that strongly co‐localized with each other in perinuclear aggregates. The p62 accumulation was time and concentration dependent after MG‐132 proteasome inhibitor loading. Interestingly, autophagy induction was p62 and Hsp70 independent. In addition, the p62 silencing decreased the ubiquitination level of the perinuclear aggregates. Recently we showed that hsp70 mRNA depletion increased cell death in ARPE‐19 cells. Here we now demonstrate that p62 mRNA silencing has similar effects on cellular viability. Conclusion The p62 and HSP70 are central molecules in the regulation of protein turnover in human retinal pigment epithelial cells in proteasome inhibitor‐ induced autophagy.
OCT features of the optic disk parameters and peripapillary nervous fibres layer thickness in children with acquired myopia
Authors: BOYCHUK IM , GORBATYUK TL 
Published: November 2010

Abstract Purpose To reveal morphological and structural peculiarities of retinal nerve fiber layer thickness (RNFLT) and optic nerve parameters in schoolchildren with acquired myopia in dependence of myopia degree. Methods 135 children (270 eyes) of 10‐14 y.o. with bilateral light (86 eyes), moderate (50) and high degree of myopia (42) as well as control group of healthy children (40) were observed. Morphostructural indices (PRNFLTh) in four segments (nasal, temporal, superior, and inferior) and optic disk parameters were investigated by optical coherent tomography SOCT Copernicus, OPTOPOL Tech.Sp.z o.o. applying standard protocols – Retina analysis and Disk analysis. Evaluation of data and comparisons in groups were performed with the help of STATISTICA for Windows 98, a Newman‐Kales criterion was applied. Results Significantly less (p=0,03) peripapillary RNFLT values were found in nasal segments in children with the high degree of myopia as compared to children in control group and with the light degree of myopia. We founded that it depended on Rim Area values. Disk Area (2,08 – 2,16) mm2 was found in 82,7% and above 3,0 mm2 in 17,3% of myopic patients. Disk Area 2,07 ‐ 4,25 mm2 was revealed in myopic patients and 2,08 – 3,11 mm2 in control group. Cup Area was 0,23‐0,54 mm 2 in myopic and 0,38‐1,13 mm2 in healthy children Rim Area 1,2‐3,2 mm2 in myopic and 1,66 – 2,63 mm2 in control group. Cup/Disk ratio 0,19‐1,18 was established in myopic and 0,19 – 1,18 in control group. Conclusion Axial myopia is often accompanied with cone near the optic disk. It is suggested that disk parameters such as – Disk Area and Rim Area can be identified as parameters which can be considered as the signs of myopia progression.
Fitting of challenging corneas with a special design of rigid gas permeable (RGP) lens
Authors: ABOU SAMRA W 
Published: November 2010

Abstract Purpose To study the safety and efficacy of a new design of RGP lens namely Rose K irregular cornea (IC) in management of corneas with high degree of irregular astigmatism. Methods The study was made in 66 eyes of 40 patients with irregular astigmatism.The selection criterion was to obtain a sample population with irregular astigmatism that was unlikely to be corrected with spectacles or conventional contact lens.A complete ophthalmologic exploration which included topography was made. All eyes were fitted with Rose K IC after measuring precisely lens parameters according to a special trial set of this design.Refraction, visual acuity before and after fitting,maximum wearing time, contact lens related problems and patient satisfaction were analyzed. Results The studied eyes included eyes with high degree of irregular astigmatism due to keratoconus in 42 eyes (63.6%),corneal injuries in 6 eyes (9.1%), PMD in 12 eyes (18.2%), and post Lasik ectasia in 6 eyes (9.1%).After fitting of the eyes with the RGP lens, there was a significant improvement in the visual acuity (VA)in all fitted eyes. The mean VA improved from 0.16±0.25 to 0.85± 0.13(p<0.001).Study of spherical and astigmatic errors showed also a highly significant improvement. The mean spherical error was ‐4.6 ± 2.4 and improved to 0.50 ± 0.20 after final fitting (p<0.001).The mean astigmatic error of studied eyes was ‐6.2 ± 2.3 while after fitting was ‐0.75±0.32(p<0.001).Statistical analysis of the subjective responses indicated a strong acceptance of the lens by all subjects. Conclusion Rose K IC proved to be effective in correcting high degree of corneal astigmatism caused by a variable number of challenging corneal conditions with high visual performance and good patient comfortability.
Ulrastructural features of keratoconus cornea after cross‐linking by riboflavin/UVA
Authors: AKHTAR S , ALMUBRAD TM , PALADINI I , MENCUCCI R 
Published: November 2010

Abstract Purpose In the present studies we assess the effects of collagen cross‐linking on ultrastructure organisation of the corneal stroma of keratoconus human corneas. Methods One normal, one keratoconus (KC) and three cross‐linked keratoconus corneas were analysed. One was treated with standard cross linking (SXL) and two with trans‐epithelial collagen cross linking (TEXL). Penetrating keratoplasty was performed three months after treatment. All samples were fixed in 2.5% glutaraldehyde containing cuprolinic blue in sodium acetate buffer and processed for electron microscopy. Results The structure of SXL corneas was very similar to normal corneas in their hemidesmosomes, basement membrane (BM), Bowman’s layer (BW) and stromal lamellae that were not undulated. The architecture of TEXL corneas presented some differences. The BM was thick with degenerated hemidesmosomes. Bowman’s layer was disorganised at some places and replaced by thin filaments forming pannus. There were thin undulating lamellae in anterior, middle and posterior stroma. The keratocytes were embedded between undulating lamellae. Large amounts of abnormal PGs were attached around collagen fibrils. The parallel running lamellae were very thin. In some parts of the anterior stroma collagen fibrils were oriented (running) in random directions instead of running parallel. There were some parts of the stroma which showed a normal appearance. Conclusion The present studies demonstrate that corneal cross‐linking leads to modifications in keratocytes and in the organisation of collagen fibril. The morphological changes might be correlated to the process of increase in biomechanical stability although there are differences between stromal structures treated by standard and trans
Corneal keratocyte population after laser subepithelial keratectomy (LASEK) with mitomycin C (MMC): 3 months vs 15 months comparison
Authors: CANADAS SUAREZ P , DRAKE P , HERNÁNDEZ‐VERDEJO JL , DE BENITO‐LLOPIS L , TEUS MA 
Published: November 2010

Abstract Purpose To study corneal keratocyte population after LASEK with mitomycin C (MMC) 15 months vs 3 months after surgery Methods Sixty eyes were included in this study, all of them treated with LASEK with intraoperative 0.02% MMC, divided into 2 groups. 32 eyes had LASEK performed 3 months before, versus 28 in the 15 months post‐op group. Keratocyte density was examined by using confocal microscopy in the anterior, mid and posterior stroma in both groups, and the mean density was compared with a control group of healthy eyes Results Keratocyte density in the anterior stroma was 16993,75+ 8001,7 cells/mm3 and 18529,1 + 2917,7 cells/mm3(p=0,3) in the 3 months and in the 15 months groups respectively, in the mid stroma it was 30783,3+ 9300 cells/mm3 in the 3 months and 20754,7 + 3615,3 cells/mm3 in the 15 months groups (p= 0,0001) and 30286,75+ 8321 cells/mm3and 19994+ 3085,9 cells/mm3 (p= 0,0001) in the posterior stroma in the 3 months and in the 15 months groups respectively. The comparison between the average densities through the whole cornea showed statistically significant difference between the 3 months vs 15 months group and 3 months vs normals, but it was not statistically significant different between 15 months vs normals Conclusion The changes in the corneal keratocyte population after LASEK + MMC seen in the early post‐op, seem to change towards the normal values 15 months after the procedure.
LASEK with MMC vs femtosecond sub‐Bowmann keratomileusis to correct myopic astigmatism: a pilot study
Authors: TEUS MA , IGLESIAS MLUZ , GIMÉNEZ C , SANCHEZ‐PINA JM , CANADAS‐SUAREZ P 
Published: November 2010

Abstract Purpose To evaluate the safety and efficacy of LASEK and Femtosecond sub‐Bowmann Keratomileusis (FSBK) in the correction of myopic astigmatism Methods Prospective, masked study of consecutive eyes, with myopic astigmatism higher than 1.5D. MMC was used in LASEK eyes if ablation > 50 microns. Visual results were evaluated three months post‐op Results 252 eyes were analyzed, 125 LASEK and 127 FSBK. Preoperative sphere and cylinder was similar in both groups (p=0.1). Safety and Efficacy indexes were 1.01 vs 1.02 and 0.90 vs 0.88 in LASEK and FSBK groups, respectively (p=0.8 and 0.3) Conclusion LASEK and FSBK achieve comparable visual results when used to correct myopic astigmatism at 3 months follow‐up
Neuroprotective effect of intravitreal cell‐based glucagon‐like peptide‐1 production in the optic nerve crush model
Authors: JONAS JB , ZHANG R , ZHANG H , XU L , MA KE , WALLRAPP C 
Published: November 2010

Abstract Purpose To examine the effect of intraocularly produced glucagon‐like peptide‐1 (GLP‐1) on the survival rate of retinal ganglion cells in an optic nerve crush model. Methods Forty‐one Sprague‐Dawley rats were divided into a study group (21 animals) in which 4 beads with 3000 genetically modified cells to produce GLP‐1 were intravitreally implanted into the right eye; a saline control group (n=12) with intravitreal saline injection; and a GLP‐1 negative bead control group (n=8) in which 4 beads with 3000 cells without GLP‐1 production were intravitreally implanted. The right optic nerves of all animals were crushed in a standardized manner. After labeling the retinal ganglion cells by injecting 3% fluorogold into the superior colliculus, the animals were sacrificed, and the ganglion cells were counted on retinal flat mounts. Results The retinal ganglion cell density of the right eyes was significantly higher in the study group (2030±470 cells/mm2) than in the GLP‐1 bead negative control group (1390±367 cells/mm2;P=0.001) and than in the saline control group (1734±424 cells/mm2;P=0.07). Correspondingly, the survival rate (ratio of retinal ganglion cell density of right eye / left eye) was significantly higher in the study group (71±19%) than in the GLP‐1 bead negative control group (49±14%; P=0.003) and than in the saline control group (57±16%;P=0.03). Conclusion Glucagon‐like peptide‐1 produced by intravitreally implanted cell beads was associated with a higher survival rate of retinal ganglion cells after an experimental optic nerve crush in rats.
Commercial interest
Interaction of vascular and biomechanical aspects of glaucoma
Authors: LESK M 
Published: November 2010

Abstract Purpose In an attempt to understand some of the reasons why some optic nerves appear to be sensitive to IOP and others not, I will review studies describing the interaction between vascular and biomechanical factors in open angle glaucoma. Methods Studies using biomechanical modelling, epidemiologic data, measurements of ocular or systemic blood flow, measurment of peripheral vasospasticity, and measurement of ocular biomechanical parameters will be reviewed. Hypotheses will be presented regarding the interpretation of these data. Results Studies suggest that the optic nerves of vasospasctic patients may be more IOP‐sensitive than those of non‐vasospastic patients. Non‐invasive measurements of ocular blood flow suggest that this pressure‐sensitivity may be related to IOP‐sensitive optic nerve blood flow. Biomechanical modelling suggests that scleral and lamina cribrosa elasticity, axial length, and eye wall thickness contribute to optic nerve head stress and strain. Cross‐sectional clinical data supports the role of increased ocular elasticity in the susceptibilty of the optic nerve to glaucoma damage, especially in vasospastic patients. Some promising new avenues for research in this area will be presented. Conclusion There is increasing evidence that biomechanical and vascular ocular factors interact leading to an elevated susceptibilty of the optic nerve to glaucomatous optic nerve damage.
Commercial interest
Development of a next‐generation sequencing platform for retinal dystrophies, with LCA and RP as proof of concept
Authors: COPPIETERS F 
Published: November 2010

Abstract Purpose Retinal dystrophies represent an emerging group of hereditary disorders that lead to degeneration of the photoreceptors and/or the retinal pigment epithelium, resulting in irreversible blindness. They are genetically complex, with over 200 disease loci identified so far. Current genetic screening consists of microarray analysis (Asper Ophthalmics) for the most recurrent mutations, and subsequent Sanger sequencing. However, the high cost and low throughput of the latter technology limits testing to only the most recurrent genes. This project aims to develop a high throughput and cost‐effective platform for screening of all known disease genes for Leber Congenital Amaurosis (LCA) and retinitis pigmentosa (RP), using the next‐generation sequencing (NGS) technology. Methods A NGS panel will be developed for all 16 and 47 known LCA and RP genes, respectively, including coding and untranslated regions, regulatory regions and microRNA binding sites. The protocol will consist of the following steps: 1) high throughput primerdesign and qPCR, 2) ligation, 3) shearing and 4) sequencing on the Illumina Genome Analyser IIx (GAIIx). This innovative protocol overcomes the need for short amplicons in order to render short‐read sequences by the GAIIx. This sequencing instrument was chosen because of its high capacity, low cost per base and the absence of interpretation problems at homopolymeric regions. Analysis of the variants will be performed using in‐house developed and commercial software, which ranks all variants according to their pathogenic potential. Conclusion Using the proposed protocol, comprehensive screening for all known disease genes for LCA and RP will be available for the first time.
Identification of novel disease gene for primary congenital glaucoma (PCG) through homozygosity mapping and next‐generation sequencing strategies in a large consanguineous pedigree
Authors: VERDIN H 
Published: November 2010

Abstract Purpose Primary congenital glaucoma (PCG) is caused by developmental anomalies of the trabecular meshwork and the anterior chamber angle resulting in an increased ocular pressure (IOP) and optic nerve damage. In general PCG displays an autosomal recessive inheritance pattern and is genetically heterogeneous. To date, three PCG loci are known, namely GLC3A, GLC3B and GLC3C, and two causal genes have been identified, CYP1B1 located in the GLC3A locus and LTPB2 located at 1.3 MB proximal to the GLC3C locus. The purpose of the current study is to identify the causal disease gene in a large consanguineous family with PCG, originating from Jordany. CYP1B1 mutations and linkage to the LTBP2, GLCB3 and GLCC3 locus were previously excluded. Methods In a first step, DNA from members from the consanguineous family will be genotyped by 250K GeneChip Mapping Affymetrix arrays. Homozygosity mapping will be applied to identify potential disease loci, using a homemade Perl script. Next, microsatellite analysis will be performed in order to confirm findings and to narrow down candidate regions. Subsequently, candidate regions of interest will be captured (Agilent) and sequenced on the Illumina Genome Analyser IIx (GAIIx). Gene and variant prioritization will be done using in‐house developed software, followed by segregation analysis and screening in control individuals. At last, a cohort of 30 molecularly unsolved PCG patients will be screened for mutations in the newly identified disease. Conclusion The identification of a new disease gene for PCG may lead to better insights into the molecular pathogenesis of glaucoma, and might uncover novel therapeutic strategies.
Role of placental growth factor (PIGF) in wound healing after glaucoma filtration surgery
Authors: VAN BERGEN T 
Published: November 2010

Abstract Purpose Failing filtering surgery due to excessive wound healing is a considerable challenge in ophthalmology, and largely contributes to progressive vision loss in glaucoma patients. Anti‐VEGF therapy helps to prevent post‐surgical scarring by inhibiting angiogenesis and collagen deposition, but does not influence inflammation (which is also an important player in postoperative wound healing). We will check the hypothesis that placental growth factor (PlGF) plays a role in scar formation after glaucoma filtration surgery, and that it may be a(n) (additional) target for improvement of the outcome of this surgery through its known anti‐angiogenic and anti‐inflammatory, and possibly anti‐fibrotic properties. Methods The effect of an anti‐PlGF antibody (ThromboGenics) will be investigated in vitro on the proliferation of endothelial cells (HUVEC), inflammatory cells (Jurkat cells) and of Tenon fibroblasts (TF). The effect of the antibody will also be investigated in vivo in a rabbit model for glaucoma surgery by measuring intra‐ocular pressure (IOP), filtration bleb function and survival, and by (immuno‐)histological analysis of angiogenesis (CD31), inflammation (CD45) and fibrosis (Sirius Red). Conclusion Our proposed research project will elucidate the potential role of PlGF‐inhibition in the improvement of filtration surgery outcome, and will highlight any angiostatic, anti‐inflammatory, and/or anti‐fibrotic effects. PlGF‐inhibition as an adjuvant anti‐inflammatory therapy to anti‐VEGF treatment in glaucoma surgery might open new perspectives for more efficient surgery. In conclusion, our project opens exciting perspectives for the treatment of the blinding condition of glaucoma, and thus might improve the visual prognosis of glaucoma patients.
Tear film biomarkers as prognostic indicators for recurrent pterygium
Authors: ZAKARIA N 
Published: November 2010

Abstract Purpose The aim of this project is to establish the use of IL‐6, IL‐8 and VEGF as biomarkers in the tear film for early detection of recurrent pterygium. Methods Cytometric Bead Array (CBA) kits will be used to perform multicytokine assays in the tear samples of eyes having pterygium before and after surgical resection of the pterygia. This technique utilizes flow cytometry to determine the concentrations of multiple analytes namely IL‐6, IL‐8 and VEGF (proposed biomarkers) present in small volumes of tear fluid.Patients with pterygia showing corneal encroachment and requiring surgical excision will be recruited in this study along with a second population of control subjects consisting of individuals with no history of eye diseases or contact lens wear. After instilling a local anesthetic, 3 drops of normal saline will be applied and the at least 50µl of the diluted epithelial secretions collected and stored at ‐80°C for CBA analysis. From the results we can determine the baseline levels of IL‐6, IL‐8 and VEGF present in normal epithelial secretions and correlate it with potentially higher levels in the eyes of patients with pterygia. By collecting post op epithelial secretions at different time points along with regular ocular surface photographs and grading of any recurrent pterygia we will be able to ascertain the role of these cytokines and growth factors as biomarkers for recurrent pterygia. Conclusion By establishing higher tear film levels of IL‐6, IL‐8 and VEGF in eyes with pterygia compared to normal eyes and the return to baseline levels post excision we can begin to ascertain the role of these key players in the pathogenesis of pterygia.
Cellular genic therapy: evaluation of mesenchymal stem cells engraftment transduced by an Ecdysone inductible lentiviral vector expressing luciferase gene
Authors: NOCHEZ Y , COLLIN C , BELLICAUD D , CARDON A , GICQUEL JJ , PISELLA PJ , GABISON E , PAGES JC 
Published: November 2010

Abstract Purpose Gene therapy might offer an opportunity to improve new treatments in ophthalmology. Several studies, using limbal stem cells or mesenchymal stem cells (MCSs), have confirmed the usefulness of these cells to repair epithelial and stromal corneal tissue. Methods We designed a strategy aimed at expressing Luciferase gene. We have tested the viability of mesenchymal stem cells modified by a lentiviral vector delivery system. To allow an adaptative treatment, the target genes would be transcriptionally controlled by the use of an Ecdysone inductible promoter. Results Construction of a lentiviral plasmid with the transgene (luciferase) and inductible promoter: Ecdysone by recombination. Construction of a lentiviral plasmid with the RXR gene and VgEcR gene and a constitutive enhancer. Transfection of 293 FT cells for lentiviral production. MSCs Transduction by “V/R” lentivirus and by “E/Transgene” lentirus. We have tested induction of Ecdysone system and transcription of the transgene (luciferase) in MSCs adding Ponasterone. Conclusion The expression of luciferase, using conditionnal expression vectors, could be applied after an engraftment of mesenchymal stem cells for pathological wound healing in Sprague‐Dawley rat cornea.