bluuebunny/crossref_metadata_2025_split huggingface dataset:
In vitro techniques for assessing differentiation and functionality of human pluripotent stem cell ‐derived RPE cells
Authors: JUUTI‐UUSITALO K 
Published: December 2011

AbstractHuman embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) serve an unlimited source of cells capable of differentiating retinal pigment epithelial cells. These cells may be used in in vitro drug testing and in cell transplantation therapies. Small population of pluripotent hESCs and iPSCs spontaneously differentiate to RPE cells. At present most differentiation methods are based on this rather inefficient method. Maturation of RPE cells from pluripotent stem cells occurs via multiple developmental phases. In those phases cells express genes and proteins that are characteristic to neuroectodermal, optic vesicle, optic cup and in the end mature RPE cells. Differentiation and maturation process of RPEs is slow, taking several weeks. However, attempts to improve the yield and the rate of maturation with biological and chemical supplements are ongoing in the laboratories around the world. The differentiation potency and maturation status is dependent on the cell line and the culture environment, furthermore hESC and iPSC derived RPE cells are also capable to trans‐differentiatiate. In addition to the basic cell and molecular biological characterizations it is vitally important to assess the differentiation and maturation status of acquired cells with functional tests. In this lecture methods to derive RPE cells from hESC and iPSC as well as functional test to assess functionality of acquired cells will be addressed.
Limbal stem cell transplantation techniques
Authors: GICQUEL JJ 
Published: December 2011

AbstractLimbal epithelial stem cells (LSC) are essential for the regeneration of the corneal epithelium. Limbal stem cell deficiency (LSCD) that can develop in traumatic, immunologic, or genetic diseases affecting the ocular surface is a major cause of corneal transparency loss.Limbal stem cell transplantation (LSCT) aims to restore a corneal epithelial phenotype, essential to maintaining the corneal transparency. The source of donor tissue containing LSC for in vivo expansion, can be the healthy fellow eye (Conjunctivo Limbal Autograft (CLAU)), living‐related (LR) Conjunctivo Limbal Allograft or cadaveric (Kerato Limbal Allograft (KLAL)). Sheets of ex vivo expanded bioengineered epithelial cells (from autologous origin (LSC / oral mucosa) or LR (LSC) are an alternative. Before and after transplantation a proper control of the ocular surface inflammation is essential. Systemic and topical immunosuppressive treatment is also needed in all allograft recipients. In order for the surgical restoration of the corneal transparency to be complete, lamellar and penetrating keratoplasty are often required.
Pathophysiology of Uveitis
Authors: DICK A 
Published: December 2011

AbstractPurpose This talk will overview the pathophysiology of non‐infectious uveitis in relation to recent SUN (standardised uveitis nomenclature) disease classification.Methods The experimental and translational human evidence of autoimmunity and activation of immunity will be discussed. In addition the talk will highlight the pathways and mechanisms of tissue damage that results in sight‐threatening disease.Results Traditionally, despite active immune regulatory mechanisms operative within the ocular environment, inflammation still occurs. Activated antigen and non‐antigen specific T cells are generated in uveitis. The interplay with innate immunity and in particular cells of myeloid lineage both systemically and within the local environment dictate the severity and extent of pathology we observeConclusion The understanding of immune responses during the uveitis open many avenues to potential novel immunotherapies that not only suppress inflammation but attempt to redress immune balance, tolerance and local homeostasis within ocular tissues
Symptoms and signs of anterior uveitis
Authors: NERI P , ARAPI I 
Published: December 2011

AbstractPurpose To review the symptoms and signs of anterior ueveitis (AU), based on the anatomic classification of uveitis, iritis and iridocyclitis.Methods Review of symptoms and signs of AU.Results Perikeratic injection, small keratic precipitates (KPs), cells and flare in the anterior chamber are peculiar findings of alternating unilateral acute non‐granulomatous anterior uveitis, which is commonly described in association with HLA‐B27 antigen and spondyloarthropaties. In such cases, hypopyon or fibrinous exudate can also occur. Patients presenting acute anterior uveitis typically show red eyes, photophobia, ocular pain, and often blurred vision. In chronic anterior uveitis, the onset is usually subtle and patients may be asymptomatic until the development of complications. Chronic flare, Koeppe and Busacca nodules of the iris, medium‐size KPs or large mutton‐fat KPs, peripheral anterior synechiae and broad‐based posterior synechiae represent hallmarks of granulomatous anterior uveitis which tends to chronicity. Viral anterior uveitis is typically unilateral, characterized by recurrent episodes of anterior uveitis. Endotheliitis, high intraocular pressure, and patchy/sectoral iris atrophy are also present. Juvenile Idipoathic Arthritis (JIA)‐associated anterior uveitis is peculiarly a bilateral non‐granulomatous chronic anterior uveitis, frequently worsened by several complications.Conclusion The typology of AU influences its clinical presentation: the clinical findings can vary on the basis of its acute or chronic, granulomatous or non‐granulomatous nature. Specific AU subtypes are characterized by a large number of distinct ocular signs.
Symptoms and signs of posterior uveitis
Authors: KHAIRALLAH M , KAHLOUN R 
Published: December 2011

AbstractPurpose Posterior uveitis (PU) is an important anatomic form of uveitis in which the primary site of inflammation is the choroid or retina, with or without subsequent vitreous involvement.Methods Review of symptoms and signs of PUResults The onset of PU can be sudden or insidious, involving one or both eyes. Most common ocular symptoms include blurred vision, loss of vision, and floaters. PU is usually associated with vitritis. Both vitreous cells and flare should be graded according to standardized grading systems. Other vitreous changes may include vitreous strands, vitreous hemorrhage, vitreous traction, and posterior vitreous detachment. Depending on the primary site of inflammation, PU can present in the form of retinitis, choroiditis, retinochoroiditis, or chorioretinitis. Retinal and/or choroidal inflammation can be focal, multifocal, or more diffuse, involving the periphery or posterior pole. It is important to distinguish between active and inactive chorioretinal disease. Retinal vasculitis can occur in the setting of several PU entities involving retinal veins or arteries. It appears as focal, multifocal, or diffuse vascular cuffing or sheathing Other retinal vasculitic changes include retinal hemorrhages, features of retinal vascular occlusion, retinal/optic disc neovascularization, and aneurysms. Macular involvement may result from direct inflammatory infiltration, macular edema, serous retinal detachment, retinal ischemia, epiretinal membrane, or macular hole. Optic nerve involvement may include optic disc hyperemia, optic disc edema, optic neuritis, neuroretinitis, optic disc exudate, and optic disc granuloma.Conclusion Clinician should be aware of the array of ocular symptoms of signs and their importance in orienting work‐up.
Diagnostic of retinoblastoma
Authors: DESJARDINS L 
Published: December 2011

AbstractPurpose Retinoblastoma is the most frequent malignant intra ocular tumors in childhood. The incidence is one out of 15000 to 18000 births. The median age at diagnosis is 24 months for unilateral and 12 months for bilateral. The genetic predisposition is autosomal doMethods Methods In 1971 Knudson made the hypothesis that 2 genetic alterations in the same retinal cell were necessary. In bilateral disease there is one germline and one somatic mutation and in unilateral non hereditary, 2 somatic mutations. The Rb1gene is located on chromosome 13 q 1‐4 . The Rb 1 protein is a pocket protein involved in the cell cycle regulationResults Results Most frequent symptoms are leukocoria and strabismus Later symptoms include heterochromia iridis,rubeosis, buphtalmia, pseudo hypopion, uveitis, inflammatory pseudo tumor and exophtalmia. Diagnosis of retinoblastoma is made by fundus examination. Imaging of the orbits and brain should be performed using MRI if possible. Ultrasonography with B and A scan is also useful as well as the use of Retcam .Differential diagnosis is sometimes easy when there is colobomas ,persistance of hyperplastic primary vitreous ,hamartomas or astrocytomas It can be difficult in cases of advanced Coats disease or when there is diffuse infiltrating retinoblastoma.Conclusion Conclusions We have made a retrospective study on patients sent for suspicion of retinoblastoma in our institute from January 2003 to December 2005 If we compare this serie to the serie published in the literature we can say that the pourcentage of well diagnosed retinoblastoma is improving. We have found 16% of wrong diagnostics. There was 30% in the serie of Balmer in 1988 and 42% in the serie of Shields in 1991. The most frequent differential diagnosis reported in all series is Coats disease
History of Disease, Facial Nerve Grading Systems & Clinical Evaluation
Authors: MAVRIKAKIS I 
Published: December 2011

AbstractObtaining an accurate history of the onset, progress and associated symptoms of newly acquired facial nerve palsy is extremely helpful in determining the potential cause of the palsy. More importantly it serves as a guide for prognosis and timing of any necessary surgical intervention. Acute versus chronic facial nerve palsy, complete versus incomplete facial nerve palsy, recovery and recurrence of the disease will be discussed. The gold standard for grading facial nerve function is the House‐Brackmann grading scale. Due to the limitations and subjectivity of this scale, several new scales of various degrees of objectivity and ease of use have been introduced. These include the Nottingham system, the Sunnybrook scale, the Yanagihara and the Sydney system, all with their advantages and disadvantages. Clinical evaluation of a patient with facial nerve palsy include evaluation of upper eyelid retraction, blink reflex, lagophthalmos, brow ptosis, paralytic ectropion, midface ptosis, mouth symmetry, platysma muscle strength, hearing, corneal sensation, Bell’s phenomenon, tear function and synkinesis.
Periocular Cosmesis & Facial Dynamic Procedures
Authors: PANTELIDIS E , LINARDOS EP , PANTELIDIS EMM 
Published: December 2011

AbstractPurpose To present Periocular Cosmesis & Facial Dynamic Procedures, that can be performed in severe and refractory facial nerve palsy cases.Methods Didactic course lectureResults Facial reanimation using dynamic procedures is a multidisciplinary rehabilitative type of management which may be undertaken in longstanding and severe facial nerve palsy. This management includes facial nerve repair, facial nerve substitution and temporalis muscle transfers. Peri‐ocular aesthetic issues need to be addressed in severe and refractory facial nerve palsy cases. Brow lift, midface lift and facial lift can be performed to improve functionality and cosmesis.Conclusion These procedures can provide a moderate to significant degree of improvement in facial nerve functionality and periocular cosmesis
Azithromycin; clinical efficacy and safety in infants
Authors: BREMOND‐GIGNAC D 
Published: December 2011

AbstractPurpose Purulent bacterial conjunctivitis affects all ages with high frequency in newborns and children. Our aim was to analyze in children especially in infants, the efficacy and safety of azithromycin 1.5% eye‐drops in the treatment of this disease.Methods Two multicenter, randomized, investigator‐masked, parallel‐group study, included infants, children and adolescents presenting a purulent conjunctivitis comparing azithromycin 1.5% eye‐drops twice daily for 3 days and tobramycin 0.3% 1 drop every two hours for 2 days then four times daily for 5 days. Cultures, signs and symptoms were studied and safety and efficacy were evaluated at baseline, Days 3 and 9.Results Results in term of positive cultures, signs and symptoms are exposed. Bacterial strains were identified precisely. Both treatments were effective. No adverse effects were noted on the ocular surface.Conclusion The distribution of positive culture in bacterial conjunctivitis in infants and children is of interest for a better understanding of bacterial conjunctivitis. A short‐treatment course can be easily used in children, thus improving the quality of life of them and their parents.
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Transduction of mesenchymal stem cells with an ecdysone inductible lentiviral vector expressing luciferase gene
Authors: FAVARD A , BELLICAUD D , NOCHEZ Y , COLLIN C , PISELLA PJ , GABISON EE , PAGES JC 
Published: December 2011

AbstractPurpose 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. However the reparation process might be hampered by a strong inflammatory reaction inducing non‐physiologic remodelling of healing tissue. The use of genetically modified cells with inducible gene to control the stromal reaction might offer an opportunity to improve new treatments in ophthalmology.Methods We aimed at transducing MSCs with an Ecdysone inducible lentivirus. To the end, we generated a conditional expression vector, expressing the luciferase under an ecdysone expression system. We have transduced MSCs and evaluated their ability to synthetize on « demand » the luciferase. Following validation of the process in cultured cells, the genetically modified cells will be evaluated in vivo, in rat.Results Using an in vitro recombination system, we generated a lentiviral vector containing the luciferase as transgene, placed under an Ecdysone promoter (« E/Luc »). To obtain an effective control of the luciferase, we generated a second vector containing the RXR gene and VgEcR gene under a constitutive promoter (« V/R »). Viral recombinant vectors were produced through 293T cells. MSCs were first transduced by the « V/R » vectors followed by the “E/Luc” lentiviruses. Finally, we have tested the responsiveness of the Ecdysone system in MSCs by adding Ponasterone and enzymatic activity of the Luc gene.Conclusion We predict that it could be applied to follow in vivo MSCs after their corneal engraftment. Upon validation of the system, the use of inducible gene expression could be applied to pathological wound‐healing.
Interleukin‐10 promotes macrophage infiltration in mouse retina
Authors: MENDES‐JORGE L , RAMOS D , VALENCA A , NACHER V , RODRIGUEZ‐BAEZA A , ALMOLDA A , CAMPBELL IL , GONZALEZ B , CASTELLANO B , RUBERTE J 
Published: December 2011

AbstractPurpose Inflammation, specifically macrophage infiltration, is known to promote a number of pathologic processes including eye neovascularization. In the current study, we explored the macrophage retinal infiltration in a mouse model that over‐expressed interleukin‐10 (IL‐10) in the retina.Methods A transgenic mouse model in C57BL6 background that overexpress IL‐10 under the control of the glial fibrillar acidic protein (GFAP) promoter was analysed. Retinas from transgenic and their wild type (wt) littermates were studied by immunohistochemistry.Results The IL‐10 transgenic mice (tg‐IL10) expressed higher levels of IL‐10 in the retina compared to wild type mice. Histologically, when compared to wt, the cytoarchitecture of the transgenic retinas did not show any differences. The only morphological alteration observed was a CD11b+ macrophage infiltration in the retina and the irido‐corneal angle of tg‐IL10. These macrophages were fully loaded with melanin. Our observations show that the number of macrophages in tg‐IL10 retinas was approximately ten fold superior than in wt. Interestingly, the 80% of macrophages in tg‐IL10 retinas were localized along blood vessels. This finding supports previous work in other paradigmas reporting infiltrated macrophages located along blood vessels, which may be involved in the modulation of blood vessel growth and regression.Conclusion A significant increase in the number of macrophages aligning along blood vessels has been observed in the retina of a transgenic model that produces IL‐10 under the control of GFAP promoter. This preliminary result suggests that IL‐10 promotes macrophage perivascular infiltration in mouse retina.
Effect of Autocrine Vascular Endothelial Growth Factor on the Survival of Retinal Müller Cells under Oxidative Stress
Authors: KIM NR , KIM CY , RHO SS , OH JH , MOON YS , CHIN HS , HONG S , SEONG GJ 
Published: December 2011

AbstractPurpose Autocrine vascular endothelial growth factor (VEGF‐A) is a known survival factor for the developing and mature retina, stimulating both endothelial and neural cells. We examined whether survival of Müller cells under oxidative stress was related to stress‐induced VEGF‐A synthesis.Methods Rat Müller cells were treated with hydrogen peroxide, and cell survivals were analyzed with a flow cytometry with annexin V‐fluorescein isothiocynate and LDH assay. The expression of VEGF‐A, VEGF‐R1, and VEGF‐R2 was determined by semiquantitative RT‐PCR or Western blot analysis.Results Hydrogen peroxide–induced cell death was increased with dose dependent manners. Treatment of hydrogen peroxide on rat Müller cells induced the expression of VEGF‐A, VEGF‐R1, and VEGF‐R2 with dose dependent manners.Conclusion In the present study, survival of Müller cells under oxidative stress was related to stress‐induced VEGF‐A synthesis. Further study is needed to reveal the protective role of VEGF‐A on retinal Müller cells.
Role of toll‐like receptor 2 and 4 of keratocytes on corneal innate immunity
Authors: CHOI HJ , LEE HJ , JEONG HJ , KIM MK , WEE WR 
Published: December 2011

AbstractPurpose To investigate the role of keratocytes in corneal innate immune system and cross‐talk of keratocytes with resident antigen presenting cells (APCs), especially through toll‐like receptor (TLR)2 and TLR4.Methods Firstly, primary cultivated C57BL/6 (B6) mouse keratocytes were stimulated by Pam3CSK4 (TLR2 agonist) or lipopolysaccharide (LPS; TLR4 agonist) and then were evaluated for their cytokine secretion. To demonstrate the cross‐talk between B6 keratocytes and APCs, cytokine changes under Pam3CSK4 or LPS challenge were assessed in co‐culture condition of B6 keratocytes with mouse dendritic cell line (DC 2.4) or mouse macrophage cell line (Raw 264.7). The reversal effect was checked out using keratocytes from TLR2 knockout (KO) or TLR4KO mice.Results Primary cultivated keratocytes from B6 mice per se actively secreted pro‐inflammatory cytokines, especially interleukin (IL)‐6, with a dose‐dependent manner in response to Pam3CSK4 or LPS challenge. With co‐culture of keratocytes with APCs, secretion of IL‐6 and tumor necrosis factor (TNF)‐α was markedly increased and it was counterbalanced by concurrent increase in IL‐10 and tumor growth factor (TGF)‐β1. After Pam3CSK4 or LPS stimulation, this cytokine balance was completely broken down by overwhelming amplification of IL‐6 and TNF‐α secretion, especially in co‐culture of keratocytes with macrophages, rather than with dendritic cells. Using keratocytes from TLR2KO or TLR4KO mice, we could find the reversal of Pam3CSK4 or LPS‐responsive dose‐dependent increment pattern in IL‐6 and TNF‐α.Conclusion These results implied that keratocytes and their TLRs could be key components for the ocular homeostasis and pathogen‐associated ocular innate immunity.
Metabolism of cholesterol in health and disease of the retina
Authors: BRETILLON L 
Published: December 2011

AbstractPurpose Cholesterol is the main sterol in the vertebrate retina.Methods Deposits of free cholesterol and cholesteryl esters at the basement of RPE are hallmarks of aging in humans, and AMD, the leading cause of vision loss in the Western world.Results Cholesterol in the neuroretina originates from in situ synthesis and extra‐retinal sources. The relative contribution of cholesterol coming from the circulation and local biosynthesis remains unknown. Tight junctions between retinal pigment epithelial (RPE) cells limit the intercellular movements of molecules to water and small molecules. RPE cells express various lipoprotein and scavenger receptors which can promote the recognition of cholesterol‐rich lipoprotein and enhance the entry of cholesterol in the neurosensory retina. Both the neurosensory retina and RPE cells express proteins which participate to cholesterol export in other tissues than the retina, including CYP46A1. CYP46A1 would represent a mechanism of cholesterol removal from neurons, by catalyzing the hydroxylation of cholesterol at position C24. Other mechanisms than CYP46A1 have been described in the retina, including formation of 7‐ketocholesterol or 27‐hydroxycholesterol.Conclusion Interestingly a single nucleotide polymorphism in cyp46a1 gene was associated with a significant risk for glaucoma, the second leading cause of blindness worldwide.
Cholesterol and neurodegeneration
Authors: PFRIEGER FW 
Published: December 2011

AbstractPurpose Based on our previous studies (Mauch et al., 2001 Science; Nieweg et al., 2009 J Neurochem), we hypothesized that retinal ganglion cells (RGCs) depend on the endocytotic import of glia‐derived cholesterol and that a breakdown of this import causes neurodegeneration (Pfrieger, 2003). The membrane protein Niemann‐Pick C1 mediates the exit of externally acquired cholesterol from the endosomal‐lysosomal system. Therefore, we studied, whether mice lacking Niemann‐Pick typ C protein1 (NPC1) due to a spontaneous mutation, show pathologic changes in RGCs and elsewhere in the retina. These mice are a model for the lysosomal storage disorder Niemann‐Pick type C.Methods We analysed the retinal phenotype of NPC1‐deficient mice by electroretinography, scanning laser ophthalmoscopy and histologic analysis.Results We observed striking signs of degeneration in retinae from two‐months‐old NPC1‐deficient mice. This included impaired visual function, accumulation of lipofuscin in the retinal pigment epithelium (RPE) layer, degeneration of photoreceptor outer segments, disruption of synaptic layers and upregulation of proteins that mediate cellular cholesterol release under the transcriptional control of the liver X receptor (Claudepierre et al., 2010 MCN). Notably, we observed an increase in LC3, an autophagy marker, specifically in the ganglion cell layer and in the RPE.Conclusion Our results support the idea that cells in the retina depend on the intercellular exchange of cholesterol via lipoprotein‐mediated transport. Moreover, our results suggest that disturbance of lipid metabolism in the retina contributes to neurodegeneration.