bluuebunny/crossref_metadata_2025 huggingface dataset:
Glucocorticoid modulation of agonist induced microvascular endothelial permeability
Authors: SHAMS F , HUDSON N , OCKRIM Z , TUROWSKI P
Published: November 2013

Purpose Conditions such as diabetic retinopathy and neural inflammation involve microvascular barrier breakdown leading to leakage, tissue oedema and immune cell influx. Glucocorticoids(GCs) have been shown to improve the barrier function of the vasculature in retina and brain. It has been proposed that GCs function through tightening of specialized junctions between endothelial cells (ECs) and reduce paracellular permeability. We investigated the effects of hydrocortisone(HC),dexamethasone(DEX),triamcinolone(TA) and a selective glucocorticoid receptor agonist(SEGRA) on microvascular endothelial permeability induced by vascular endothelial growth factor(VEGF),lysophosphatidic acid(LPA) and histamine(Hist), all of which are proposed to be involved in diabetic and inflammatory neurovascular pathologies.

Methods In vitro studies were undertaken in cultures of primary rat brain and retinal microvascular ECs, as well as a novel immortalised rat retinal EC line, PT2. Junctional protein characterisation was performed by indirect immunocytochemistry and confocal microscopy. Permeability was measured by macromolecular flux assays.

Results Primary retinal and cerebral EC cultures exhibited an exquisite apico‐basal polarity in their response to vasoactive compounds, maintained high barrier properties and sophisticated junctional protein complement. Passaged or immortalised EC had lost most of these features. HC and DEX were effective in suppressing VEGF‐, LPA‐ and Hist‐induced permeability. TA results were similar, except Hist‐induced permeability was insensitive to TA. SEGRA was ineffective in preserving microvascular barrier function.

Conclusion The effectiveness of GCs depends on the pathology involved in particular the vasoactive substance in play.
Natural history of vitreoretinal interface disorders
Authors: POURNARAS JA
Published: November 2013

Natural History of main vitreoretinal interface disorders as epiretinal membranes, macular holes, vitreomacular traction syndrome (VMTS) and myopic foveoschisis will be discussed.
Medical management of vitreoretinal interface disorders
Authors: LE MER Y
Published: November 2013

Year 2013 will be a cornerstone in the history of therapeutic vitreolysis with the European approval of the ocriplasmine in March, the first commercially available product to achieve in some selected cases a separation of the vitreo‐retinal interface. Since the publication in 1993 of the first vitreous detachments in rabbit eyes, obtained by intravitreal injection of plasmine, the road has been opened to several researches looking for the best candidate to produce this detachment in human eyes. The published data of the use of ocriplasmine to date are still too fragmentary to give a precise idea of what could be the best indications to perform an intravitreal injection of drug instead of a classical vitrectomy. Aside the symptomatic vitreo‐macular tractions syndrome and small idiopathic macular holes, some other disorders in which an abnormal vitréo‐retinal adhesion seems to play a role are currently evaluated, such as the early stages of macular degeneration and diabetic macular edemas. We’ll present the last available data, focusing beyond the pure scientific results, on the possible consequences on our everyday surgical practice and the clinical decision tree in vitreo‐retinal disorders.

Commercial interest
Surgical management of vitreoretinal interface disorders
Authors: WOLFENSBERGER TJ
Published: November 2013

Purpose: To assess safety and efficacy of novel surgical therapies for vitreomacular interface disorders Background: Vitreomacular interface disorders are characterised by a combination of mechanical and biochemical interactions between the posterior hyaloid and the internal limiting membrane and the retinal surface in general. Pathologies that are amenable to surgical treatment include primarily epiretinal membranes and macular holes, but other less common maculopathies have been come to the forefront recently. Results: This presentation will summarise the latest developments in the surgical therapy of vitreomacular interface disorders by evaluating the most recent data on advances in diagnostic tools for preoperative decision making, surgical technology and pharmacological combination therapies. Conclusion: Novel surgical techniques are constantly increasing the therapeutic succes while reducing tissue irritation during surgery. New diagnostic approaches including high‐resolution OCT can help the decision making and to predict visual acuity recovery after vitreomacular interface surgery.
Case study
Authors: POURNARAS JA
Published: November 2013

Case Study will be proposed and therapeutical options will be discussed with the panel.
Tthe role of low‐grade inflammation in glaucoma
Authors: KOLKO M
Published: November 2013

Glaucoma has been associated with low‐grade inflammation. The exact role of inflammatory processes in the pathophysiology of various types of glaucoma is, however, unclear. In addition, few in vivo studies have provided direct evidence for inflammatory processes in glaucoma. Disturbed auto regulation has been shown to increase the levels of Endothelin‐1 (ET‐1) and/or nitric oxide (NO) thereby causing inflammation by either inducing ischemia or oxidative stress. Evidence has furthermore revealed a diminished Müller cell ability to remove excess glutamate in the synaptic cleft in response to vascular dysregulation. This in turn causes glutamate excitotoxicity with a hyperstimulation of the NMDA receptor. Activation of NMDA‐receptors has been shown to induce TNF‐α production in the Müller cells, indirectly causing RGC death through TNF‐α stimulation of TNF‐α receptor 1 on RGC. Specifically, this aspect of RGC death rise the possible involvement of low‐grade inflammation as an important factor in the metabolic and excitotoxic intradependance between Müller cells and RGC. The current knowledge of low‐grade inflammatory changes in glaucoma and ideas on possible future targets for pharmaceutical intervention in glaucoma will be discussed
Mohs surgery of eyelid tumours assisted by ex vivo confocal microscopy and a new “open book” method for tissue preparation
Authors: GRIVET D , CAMPOLMI N , CINOTTI E , FOREST F , ESPINASSE M , PERROT JL , LABEILLE B , CAMBAZARD F , DUMOLLARD JM , PEOC'H M , GAIN P , THURET G
Published: November 2013

Purpose The micrographic Mohs surgery, that allows a complete and tissue‐economic resection of skin tumours, is particularly adapted to eyelid margin tumours (EMT) surgery. However, being time consuming for pathologists it is not used in routine. Aim: to expose a fast and reproducible method of Mohs surgery for EMT associating ex vivo confocal microscopy (EVCM) and an original tissue preparation named “open‐book”

Methods EMT were surgically resected, let unfixed and unstained, immediately prepared with the “open book” technique and analysed with EVCM (vivascope 2500, Lucid Inc, NY and MAVIGH GmbH, Germany). The “open book” technique consisted of splitting the eyelid in two lamellae maintained by a hinge situated at the eyelid margin and flat‐mounting it between two glass slides in order to allow acquisition of both cutaneous and conjunctival sides in the same EVCM scans. The whole sample volume was scanned at the 3 wavelengths available (488, 658 and 785 nm) and the mosaics was automatically reconstituted by the built‐in software. Tumours were then immediately fixed in 4% PFA and a classical pathologic analysis was performed. Agreement between tumour limits found in EVCM analysis and in classical pathology was calculated with weighted kappa coefficient

Results In all cases, the skin and conjunctival tumours limits determined by EVCM were comparable to classical pathology

Conclusion ECVM combined with original “open book” tissue preparation could take a place in routine of eyelid tumour surgery. Grant: GIRCI jeune chercheur Rhone Alpes Auvergne 2013 No financial interest for all authors
Anatomical and visual outcome after pars plana vitrectomy in acute postcataract endophthalmitis
Authors: CHIQUET C , CAMPOLMI N , COMBEY DE LAMBERT A , CORNUT PL , ROUBEROL F , PALOMBI K , APTEL F , THURET G , CREUZOT‐GARCHER C
Published: November 2013

Purpose To report visual and anatomical outcome of patients vitrectomized for acute post bacterial endophthalmitis

Methods 123 patients with acute postcataract endophthalmitis and consecutively treated by pars plana vitrectomy (PPV) were included in four academic hospitals (French Institutional Endophthalmitis Study (FRIENDS) group).

Results At 6 month follow‐up, 49 patients (40%) had a visual acuity (VA) greater than or equal to 20/40, and 83 patients (68%) had VA greater than or equal to 20/200. Baseline factors associated with final VA ≥20/40 were a younger age (Odds ratio, OR= 1.05 (1.05‐1.09), p=0.006), presence of fundus visibility (OR=7 (1,4‐34,6),p=0.007), and absence of cataract surgery complications (OR=15.3 (1.9‐32), p=0.001), corneal edema (OR= 2.3 (1.1‐4.9), p= 0.02), hypopion (OR= 2.6 (1.1‐6.6), p=0.04), or of virulent bacteria (OR=3.1 (1.4‐6.7), p=0.005). At the time of PPV, absence of vasculitis at the posterior pole at the time of PPV (OR= 7 (1.4‐34.6), p=0.02) and a lower duration of PPV (OR=1.02 (1.01‐1.04), p=0.02) were significantly associated with final VA ≥20/40. Risk factors of RD were diabetes (OR = 4.7 (1.4‐15.4), p=0.01), and visualization of retinal vasculitis on the posterior pole (OR = 3.8 (1.1‐13.9), p= 0.03) at the time of PPV.

Conclusion PPV allowed to be beneficial in a majority of the patients. Baseline inflammatory signs and bacterial virulence are the main prognostic visual factors. RD remains the major complication but final anatomical and visual outcome is still poor, despite updated vitreoretinal techniques
Genetic analyses of autosomal optic neuropathies reveal novel physiopathological pathways involved in optic nerve degeneration
Authors: LENAERS G , HALLOY F , HIEU N , MEGY C , DELETTRE C , MEUNIER I , ROUBERTIE A , HAMEL CP
Published: November 2013

Autosomal Optic Neuropathies (AON) are rare mitochondrial diseases affecting the retinal ganglion cells that form the optic nerve and transduce the visual informations from the retina to the brain. They are heterogeneous inherited diseases with dominant – the most abundant ‐ and recessive forms, for which only three genes have so far been identified, OPA1, OPA3 and OPA7. Together, mutations in these genes account for less than 50% of patients with an ascertained clinical diagnosis, OPA1 being by far the most frequent gene involved. To gain insight in the genetic diagnosis of AON, we performed whole exome sequencing on dominant and recessive families, for which we excluded mutation in OPA1, 3 and 7. Results identified some novel genes, all encoding mitochondrial matricial proteins. Importantly none encodes protein embedded in the inner mitochondrial membrane, as are OPA1, 3 and 7, but all have a function related to the lipid and protein composition of this membrane. Thus these results put emphasis on novel pathophysiological mechanisms involved in retinal ganglion cells degeneration, again pointing on the absolute requirement to maintain the inner mitochondrial integrity to preserve optic nerve function and survival.
OPA1 in mitochondrial quality control and its implications for RGC degeneration
Authors: ALAVI M
Published: November 2013

Mitochondria are vulnerable to damages of their proteins, lipids, and mtDNA by various stress factors and mitochondria either recycle dysfunctional mitochondria via the mitochondrial autophagy pathway, arrange for the recycling of the complete host cell by apoptosis, or continue to function normally and supply the host with energy and metabolites. This is called mitochondrial quality control and it is crucial for all neurodegenerative diseases, including the most prominent ones, Alzheimer’s disease and Parkinson’s disease. Mitochondrial quality control is facilitated by mitochondrial network dynamics – continuous fission and fusion of mitochondria. Mitochondrial fission is accomplished by DRP1. MFN1 and MFN2 on the mitochondrial outer membrane, and OPA1 on the mitochondrial inner membrane are necessary for mitochondrial fusion. Mitochondrial network dynamics are regulated in highly sophisticated ways by post‐translational modifications and protein processing. I would like to present selected aspects of mitochondrial quality control using OPA1 as an example and discuss their implication on Dominant Optic Atrophy, a slowly progressive optic neuropathy with juvenile onset caused by mutations in the OPA1 gene.
Hypothermic storage or organ culture?
Authors: PONZIN D
Published: November 2013

Currently there are two storage practices for the cornea, both use liquids based on cell culture medium: the hypothermic storage at 2‐6°C, and organ culture at 30‐37°C. Hypothermic storage seems to offer donor tissues of good quality comparable to that obtained by organ culture, provided that the storage time is kept short. Indeed, according to the literature, the risk of primary graft failure increases significantly after storage longer than 7 days. Furthermore, corneas stored longer than 7 days display epithelial alterations that may hinder the surgical procedure or delay the full recovery of the graft. The organ culture storage method consists of a storage period in culture medium at 30‐37C°, and a shorter de‐swelling and transportation phase at 30‐37C° and room temperature in the same medium supplemented with 4‐8% dextran. Organ culture solutions contain fetal calf serum as a source of growth factors to limit endothelial cell loss. The serum must be obtained from prion‐free countries. A storage period of 30 days can be achieved without significant loss of endothelial cells. The endothelium shows reparative phenomena during storage. Organ culture offers a longer storage time, a less restricted donor supply on beforehand, corneal endothelium with a better defined quality, and a pre‐operative microbiologic control. Cultured corneas always display an epithelium made up of 2‐3 layers of viable cells. The 30 days storage period allows an efficient use of valuable donor tissue: planning of operations is easier, allowing sufficient time for the allocation. The disadvantages of this method are the relative technical complexity and the need for qualified staff to perform tissue culture and selection of the corneas. The studies comparing the effect of the storage methods on outcome demonstrate similar graft survival and post‐operative decline in endothelial cell density. Corneas stored by organ culture are at least comparable with those stored hypothermically for shorter periods.
Intravitreal steroids to treat the RVO‐associated macular edema
Authors: CREUZOT C
Published: November 2013

Intravitreal steroids are interesting tools to treat macular edema associated to RVO. The effect of the treatment lasts approximately 4‐5 months. The main risk is increased ocular pressure ranging from 15% to 50% according to the type of steroids. Patients with glaucoma treated with at least two medications or unbalanced glaucoma will be treated with another therapeutic option to avoid complications. The regular follow‐up of the patient will define efficacy and safety of the treatment. For re injections, the rythm of control can be slightly eased.

Commercial interest
Controversies in the surgical treatment of RVO ‐ Associated macular edema
Authors: GARCIA ‐ ARUMI J , VELáZQUEZ VILLORIA D
Published: November 2013

Macular edema is the main cause of decreased visual acuity in retinal vein occlusions (RVOs). Most approaches to the management of RVOs, such as grid macular laser photocoagulation and intraocular injections of drugs, are addressed to reduce the permeability of the macular vascular net and to increase the vitreoretinal fluid exchange of oxygen and protecting factors. However, we need to solve the primary mechanism of the disease: the vein occlusion. In order to reperfuse the thrombosed vein, Radial Optic Neurotomy (RON) is a surgical procedure aimed at solving the “compartment syndrome” that may exist in eyes with CRVO or hemicentral RVO. Sheathotomy specifically releases the compressive factor at the arteriovenous crossings by sectioning the adventicial seath. Recombinant tissue plasminogen activator (r‐tPA) has been administered by several routes, including systemic, intravitreal and by endovascular cannulation of retinal vessels. PPV with peeling of the internal limiting membrane has also been suggested. Surgery remains a good treatment option for patients with very recent and edematous occlusion with poor visual acuity, as reperfusion of the vein is the best approach to treat the macular edema and avoid ischemic complications.
Pseudoexfoliation: normative data and associations. The Central India Eye and Medical Study
Authors: JONAS J , NANGIA N
Published: November 2013

Purpose To assess the prevalence of pseudoexfoliation (PEX) and its associations in a population‐based setting.

Methods The population‐based Central India Eye and Medical Study included 4711 individuals. All study participants underwent a detailed ophthalmological examination. After medical pupil dilation, PEX was assessed by an experienced ophthalmologist using slit‐lamp based biomicroscopy.

Results PEX was detected in 87 eyes (prevalence: 0.950.10% (95%CI:0.75,1.15) of 69 subjects (prevalence: 1.490.18% (95%CI:1.14,1.83). In multivariate analysis, PEX prevalence was associated with higher age (P<0.001; OR:1.11), lower body mass index (P=0.001) and higher diastolic blood pressure (P=0.002). In the multivariate analysis, PEX was not associated with retinal nerve fiber layer cross section area (P=0.76) and presence of open‐angle glaucoma (P=0.15). Side differences in the presence of PEX were not significantly associated with side differences in intraocular pressure (P=0.40).

Conclusion In a rural Central Indian population aged 30+ years, PEX prevalence (mean: 1.490.18%) was significantly associated with older age, lower body mass index and higher diastolic blood pressure. It was not significantly associated with optic nerve head measurements, refractive error, any ocular biometric parameter, nuclear cataract, early age‐related macular degeneration and retinal vein occlusion, diabetes mellitus, smoking, and dyslipidemia.
Dependence of the origin of pain in the eyes with terminal stage of glaucoma from chemical composition of vitreous
Authors: ERMOLAEV A , RENDEL E , KASCHEEVA N
Published: November 2013

Purpose To identify a possible link between the origin of hypertonic pain syndrome (HPS) in the eyes with terminal stage of glaucoma and the difference of the urea concentrations between vitreous contents and blood serum.

Methods 31 patients (31 eyes) with refractory terminal glaucomas of different origins with IOP more 35 mm Hg were studied and operated on. The principle of surgery was drainage of the vitreous space for HPS elimination. Some vitreous contents and venous blood were taken during the surgery and researched for measuring urea concentrations. The samples were centrifuged then the blood serum and the supernatant of the vitreous contents were analyzed (SPOTCHEM‐EZ SP 4430). The patients were divided into 3 groups depending on the intensity of pain syndrome: 1st with strong pain; 2nd ‐ pain was present earlier; but it was absent before the surgery; 3d ‐ pain was never registered.

Results A high concentration of urea in vitreous contents was found. Only in 6.45% the urea concentration in blood serum was higher then in vitreous. In 12.9% concentrations were near equal and in 80.64% the urea concentration in vitreous contents was higher than in blood serum. Pain existed in the cases when the difference (the negative gradient) of urea concentrations exceeded 6.1 mmol/L only.

Conclusion High concentration of the osmoticaly active urea in the vitreous cavity can be the reason for increase of IOP in some cases of refractory glaucomas. If the negative gradient of the urea concentrations were higher than the critical level, it would be the reason for the development of HPS.
Should our decision to practice cataract surgery be only based on visual acuity?
Authors: VAN DEN BERG TJTP
Published: November 2013

An obvious minimum criterion for cataract surgery is the likelihood that the surgery will be beneficial to visual function of the patient. Visual function is in practice often simplified to visual acuity VA. Although, most surgeons agree that VA is the most important parameter, it is also agreed there are other visual function factors to consider, corresponding to complaints such as glare, hazy vision, face recognition problems. This may suggest a complicated multidimensional array of visual functions to be important. To address this question in a comprehensive way, the functional point‐spread‐function is useful. The effect of cataract can be summarized by its small angle effect, dominating VA (and contrast sensitivity), and its large angle effect, corresponding to straylight SL. The question then is how important this large angle domain of visual function is for the cataract patient. Results of 2 studies will be presented, a questionnaire study in cataract patients, and a study of referral practice for CE. Subjective complaints were documented before and after surgery by the 39‐item National Eye Institute Visual Function Questionnaire (NEI VFQ‐39). Both studies show dominance of VA, but with an important second place for SL.

Commercial interest
The effectiveness of wavefront‐guided refractive laser treatment
Authors: SULEMAN H , FARES U , AL‐AQABA MA , OTRI AM , SAID DG , DUA HS
Published: November 2013

PURPOSE: To compare the efficacy, predictability, safety and induced higher‐order aberrations (HOAs) between wavefront‐guided and non‐wavefront‐guided ablations. METHODS: The Cochrane Central Register of Controlled Trials, PubMED, and EMBASE were searched for randomized controlled trails. Trials meeting the selection criteria were quality appraised and data extracted by 2 independent authors. Measures of association were pooled quantitatively using metaanalytical methods. Comparison between wavefront‐guided and non‐wavefront‐guided ablations was measured as pooled odds ratios (ORs) or weighted mean differences. The pooled ORs and 95% confidence intervals (CIs) were computed for efficacy, safety, and predictability. The weighted mean difference and 95% CIs were used to compare induced HOAs. RESULTS: Eight trials involving 955 eyes were included. After wavefront‐guided LASIK, the pooled OR of achieving uncorrected distance visual acuity (UDVA) of 20/20 (efficacy) was 1.10 (95% CI, 0.66‐1.83; P=.72), the pooled OR of achieving a result within ± 0.50 diopter of intended target (predictability) was 1.03 (95% CI, 0.60‐1.75; P=.92), and the weighted mean difference in induced HOAs was ‐0.09 (95% CI, ‐0.17 to ‐0.01; P=.04). No study reported loss of 2 or more lines of Snellen acuity (safety) with either modality. CONCLUSIONS: Metaanalysis showed no clear evidence of a benefit of wavefront‐guided over non‐wavefront‐guided ablations. With high preexisting HOAs, however, wavefront‐guided has advantages over non‐wavefront‐guided treatment.
Tips and tricks in grossing & processing specimens
Authors: COUPLAND S
Published: November 2013

Ophthalmic specimens range in size and degree of complexity. The diagnostic work‐up of ocular‐, ophthalmic biopsies and larger specimens demands close collaboration between the clinician, the pathologist and other specialties, including the vitreoretinal surgeon, plastic surgeon, as well as the head‐and‐neck surgeon. Documentation of all relevant clinical information in the pathology request form as well as timely discussions between the various specialists (e.g., telephone or E‐mail communications prior to a biopsy being performed) are essential components of the diagnostic and referral pathways. The laboratory itself should be equipped with experienced technical staff who are dedicated to eye specimens and familiar with the specimen protocols. Further, a pathologist with expertise in ocular pathology/fluid samples should be supported with a wide range of investigations, including molecular diagnostic techniques. In this way, the yield from these samples can be optimized to reach an unequivocal diagnosis, rapid communication to the clinician, and timely instigation of therapy. Guidelines for these steps are provided.
Paraneoplastic syndromes
Authors: SZATMARY G , POLGAR TATJA
Published: November 2013

Neuro‐ophthalmologic disorders in the patient with cancer Paraneoplastic syndromes Background: Paraneoplastic syndromes are caused by remote effect of cancer rather than direct invasion. Objective: Review of paraneoplastic neuro‐ophthalmic manifestations, differential diagnosis, diagnostic workup and proposed pathophysiology will be presented. Methods: Literature review along with representative case presentations. Results: Overall incidence of remote effect of cancer is 10‐15 % of all cancers. The number of autoantibodies in association with paraneoplastic syndromes is accumulating. Conclusion: Paraneoplastic syndrome with ocular manifestation may be the first sign of malignancy, and therefore ophthalmologist may play a crucial role in early detection and timely treatment with potential improvement of the disease course.
Radiation optic neuropathy
Authors: BORRUAT F
Published: November 2013

Radiation optic neuropathy (RON) is a radionecrosis of the anterior visual pathway. It is a rare delayed and unpredictable complication of radiotherapy. RON presents with an acute painless and permanent visual loss. Both accurate and early diagnoses are mandatory as some specific therapies might benefit the patient. This presentation will summarize the physiopathology of RON with an emphasis on clinical diagnosis and therapeutic possibilities.
High resolution retinal image analysis of unilateral retinitis pigmentosa using adptive optics
Authors: GOCHO K , KIKUCHI S , KABUTO T , AKEO K , KAMEYA S , YAMAKI K , TAKAHASHI H
Published: November 2013

Purpose To report the findings of en face adaptive optics (AO) fundus imaging in eyes with unilateral retinitis pigmentosa(RP) patient.

Methods The both eyes from a unilateral RP patient, 52 y.o. female, was imaged using both an infrared AO camera (rtx1™, Imagine Eyes, France) and spectral‐domain optical coherence tomography (SD‐OCT – Cirrus™, Carl Zeiss Meditec, Germany). Full field scotopic and phototic electroretinograms(ERGs) were recorded according to the ISCEV standard. Normal funduscopic examination, Goldmann visual field and fluorescence angiography(FA) were also examined. The AO images acquired from the unilateral RP patient were analyzed on cone density and then compared to the AO data from the eyes of 21 healthy volunteers with a mean age of 36 years (range 20‐57).

Results The unilateral RP patient showed boney spicule changes, vascular attenuation and decreased peripheral visual field in the right eye. FA showed granular hyperfluorescence in the post pole, mottling of RPE and atrophy of choroid capillary in the peripheral retina in the right eye. Her ERGs of the right eye showed non‐recordable in all responses tested. OCT findings in the right eye showed preservation of the photoreceptor layer only in the foveal and parafoveal regions with loss in the more peripheral macula. All these results in her left eye were normal. AO data showed severe reduction of cone density in the right eye and the left eye showed normal cone density and regular cone mosaic.

Conclusion In unilateral RP patient, AO showed only one eye was affected even in the cellular level examination. The examinations using AO may be useful in diagnosis and better understanding of pathology and management of unilateral RP cases.
Pilot study in Polish keratoconus patients using next generation sequencing
Authors: GAJECKA M , KAROLAK JA , NOWAK DM , POLAKOWSKI P , SZAFLIK JP
Published: November 2013

Purpose Next generating sequencing (NGS) is a useful tool in disease etiology evaluation. Keratoconus is a multifactorial disease in which NGS technology may be applicable. Here we present a design of keratoconus study based on transcriptome analyzes in human corneas derived from keratoconus and control individuals from Poland.

Methods A total of 96 individuals from Polish population are included into this study: 48 keratoconus patients and 48 individuals without keratoconus phenotype. Illumina HiScan SQ system is applied in transcriptome analyzes in the study.

Results Total RNA from corneas for RNAseq and small RNA analyzes was extracted. Transcriptome analyzes were initiated.

Conclusion Since genetic factors involved in keratoconus etiology are in majority unrecognizable, the newest available technologies are necessary to be involved in further keratoconus research. Support: Polish Ministry of Science and Higher Education, Grant no. NN402591740 and National Science Centre in Poland, Grant no. 2012/05/E/NZ5/02127.
Role of O‐GlcNAcylation of Sp1 in the pathogenesis of diabetic retinopathy
Authors: DONOVAN K , ALEKSEEV O , AZIZKHAN‐CLIFFORD J
Published: November 2013

Purpose Central to the pathogenesis of diabetic retinopathy (DR) is vascular endothelial growth factor A (VEGF‐A). Gene expression of VEGF‐A is regulated by several transcription factors, including specificity protein 1 (Sp1). Sp1 is known to participate in angiogenesis and co‐localizes with VEGF‐A in the epiretinal membranes of DR patients. Sp1 is heavily O‐GlcNAcylated by the enzyme O‐GlcNAc transferase (OGT). This glycosylation modification affects its transcriptional activity, localization, stability, and protein interactions. Since O‐GlcNAcylation of proteins is elevated under high glucose conditions, we investigated the relationship between O‐GlcNAcylation of Sp1 and increased VEGF‐A transcription.

Methods Hyperglycemia‐exposed ARPE‐19 (retinal pigment epithelial cells) and TR‐iBRB (rat retinal microendothelial cells) were assayed for levels of VEGF‐A by RT‐qPCR and Western blot. VEGF‐A protein and mRNA levels were measured in cells depleted of Sp1 or OGT by shRNA. Small molecule inhibitors of OGA and OGT were used to increase or decrease total O‐GlcNAc. Chromatin immunoprecipitation was used to assess levels of Sp1 bound to the VEGF‐A promoter.

Results Hyperglycemia caused increased VEGF‐A transcript and protein expression in ARPE‐19 and TR‐iBRB cells. OGT or Sp1 depletion significantly abrogated these glucose‐induced changes in both cell types, while minimally affecting basal expression. ChIP analysis showed that glucose increased the amount of promoter‐bound Sp1.

Conclusion Glucose‐induced increases in pan O‐GlcNac in the RPE and vascular retina may be participating in the aberrant expression of VEGF‐A in DR. The highly O‐glcosylated Sp1 transcription factor may be responsible for early VEGF‐A production in the retina.
Genotype‐phenotype correlation in two patients with posterior polymorphous corneal dystrophy 3
Authors: LISKOVA P , PALOS M , HARDCASTLE A , VINCENT A
Published: November 2013

Purpose To determine the molecular genetic cause of posterior polymorphous corneal dystrophy (PPCD) in four Czech probands.

Methods Extensive ophthalmological examination including Pentacam and specular microscopy, and direct sequencing of the ZEB1 coding region were performed.

Results Two novel frameshift mutations within ZEB1 were identified; c.2617dup in exon 8 in a 21‐year‐old female, considered to be most likely de novo in origin, and c.698dup in exon 6 in a 21‐year‐old male. The first case had mild changes consistent with a PPCD diagnosis and best corrected visual acuity (BCVA) bilaterally was 1.0. The corneal phenotype of the second case was more severe with a BCVA of 0.4 in the right and 0.05 in the left eye. Corneas of both probands were abnormally steep (keratometry readings K1 ≥ 47.4 D and K2 45 ≥ 49.2 D) with increased pachymetry values, but no pattern indicative of keratoconus. Specular microscopy in both patients revealed reduced endothelial cell density (range 1055‐ 655 per mm2). Both probands had a history of surgery for inguinal hernia; the male patient also reported the presence of hydrocele.

Conclusion Nucleotide changes within the coding region of ZEB1 underlie the pathogenesis of PPCD in 4 out of 23 (17.4%) Czech probands.
The role of wound healing in cornea
Authors: BEUERMAN R
Published: November 2013

Corneal fibrosis contributes to the substantial health burden due to “corneal blindness”, a leading cause of blindness world‐wide. Molecular intra‐cellular regulators of the cytoskeleton, suggests that this class of proteins may be targets for therapeutic development. In the transformation the usually quiet fibroblast‐to‐a myofibroblast morphology, the expression of α‐smooth muscle actin (SMA) is characteristic. As an excessive healing response, fibrosis is the result of the corneal keratocyte to myofibroblast transformation and may occur in response to trauma, infection as well as surgery. Using a mouse model of fibrosis following both infection with Pseudomonas and by trauma (an anterior keratectomy) it was shown using an array for the cytoskeletal regulators that one key molecule was highly unregulated. Moesin a member of the ERM Complex (ezrin/radixin/moesin) was highly unregulated. The use of siRNA applied by iontophoresis showed that the molecules characterizing fibrosis, such as alpha‐SMA. The clinical development focuses on a method that can be applied topically to lessen or prevent fibrosis after corneal insult.
The mouse model of glaucoma filtration surgery: from understanding the wound healing response to applications as a tool for validating anti‐fibrotics
Authors: LI‐FONG LF , WONG TT
Published: November 2013

Purpose: Failure after glaucoma filtration surgery (GFS) is attributed to fibrosis at the operated site. We have developed a mouse model for GFS so as to understand the wound healing process after this procedure and to test anti‐fibrotics for future clinical application. Methods: Modified GFS was performed on mice and bleb survival was determined by slit lamp photography. Real‐time PCR, antibody‐based cytokine assays, flow cytometry, and immunofluorescent analyses were performed to determine gene and cellular changes. Results: The mouse model of GFS mimicked the clinical response to mitomycin C. Further analysis of the operated tissues revealed that the post‐surgical wound healing response was separable into two distinguishable phases: an early “acute inflammatory” phase followed by a late “fibrotic” phase. Conclusions: Our data suggest that targeting the distinct phases of the wound healing process after GFS may be desirable in order to overcome the long term scarring response. Furthermore, the mechanisms of potential anti‐fibrotics may now be understood in terms of their specific effects on each phase of the wound healing process. Paper presentation at Anatomy & Cell Biology, Special Interest Symposium.
Dry eye associated with ocular surface damages
Authors: SCHMETTERER L
Published: November 2013

Abstract not provided
Choroidal thickness measurements and variations with age, gender in normal, myopia and high myopia
Authors: COSCAS G
Published: November 2013

To evaluate CT and variations along foveal horizontal scan and age related change in CT in normal and myopic Methods:Multi‐center prospective study with 3 groups:30 Normal,24 Mild Myopia,31 High Myopia. Manual measurement followed choroid boundary selection(supra‐choroid space,lamina fusca and outer limit of vessels. CT was measured at 500 µm intervals up to 1,500 µm temporal and nasal, manually by two graders independently. Statistical analysis evaluated CT at each location,the effects of age and myopia and grader agreement.Results: 85 eyes were qualified.Excellent grader agreement:intra‐class correlation>0.97(Bland‐Altman analysis).The mean CT was 248.2±78.5µm for Normal‐‐age=58±18;247.0±85.4 for Myopia‐age=45±20;131.5±70.9for High Myopia‐age=54±13. The mean CT was significantly thicker for Normal and Myopia than High Myopia. CT was highly correlated across all test locations and slightly thicker underneath the fovea. The overall slope of age related mean change for the mean CT was ‐1.95 µm/year.The effect of age differed among the study groups.CONCLUSION: OCT measurements of CT are reproducible.Age and high myopia negatively affect CT.There was good agreement in CT measurement between RTVue and Spectralis.
Choroidal circulation measurements by laser Doppler flux metry
Authors: POURNARAS CJ
Published: November 2013

Continuous real time Laser Doppler Flowmetry (LDF), has been used in humans, for the evaluation of the subfoveal choroidal circulation and the investigation of the regulatory processes of the choroidal blood flow in response to various physiological stimuli, as increases and decreases of the ocular perfusion pressure. A progressive decrease of the choroidal blood flow was observed with the aging and the progession of the age related macular degeneration. Abnormal regulation of the choroidal blood flow in patients suffering with neovascular AMD or Diabétic microangiopathy were also reported. The correlation of the subfoveal choroidal blood flow with the anatomical changes of the choroidal circulation as observed by OCT are under investigation.
Evaluation of posterior limits of choroid
Authors: UZZAN J
Published: November 2013

Purpose: Recent studies have reported choroidal thickness measurements based on FD‐OCT images. Lamina fusca (LF) is the natural anatomical outer boundary of the choroid. Unfortunately, LF is not always clearly visible in OCT images. The goal of this study is to assess the difference between LF boundary and posterior choroidal vessel (PCV) boundary. Methods: In a retrospective case review study, Horizontal B‐Scan images, through the center of the fovea were reviewed and 96 eyes were included. The eyes were divided into 3 categories based on the characteristics of the choroid‐sclera junction: SCS if suprachoroidal space is visible as a dark strand, LF if lamina fusca is visible as a bright strand, and PVC if only posterior vessel cavity boundary is visible. Results: For the 48 eyes in either SCS or LF, choroid thickness measurement differ by 26.9μm on average, ranging from 8 to 85 μm in individuals between the two boundaries. Conclusions: Choroid thickness measurement could be affected by boundary criteria. Posterior vessels boundary is the best method in 50% of cases but the presence of Supra Choroidal Space must be remembered. A common posterior boundary definition may be necessary to avoid significant bias in further studies.
Comparison of choroidal thickness between central serous choroidopathy and polypoidal choroidal vasculopathy
Authors: PUCHE N , COSCAS F
Published: November 2013

PURPOSE: To compare choroidal thicknesses among eyes with polypoidal choroidal vasculopathy (PCV), and central serous chorioretinopathy (CSC). METHODS: Patients with PCV and CSC underwent enhanced depth imaging optical coherence tomography (EDI OCT). A horizontal linear section comprising 100 averaged scans was obtained of each macula. The choroidal thickness was measured from the outer border of the retinal pigment epithelium to the inner scleral border. Seventeen healthy subjects (17 eyes) also underwent EDI OCT. RESULTS: A total of 17 eyes of 17 consecutive patients with CSC (17 male, mean age 50 years range 37‐66 years) and 15 eyes of 15 patients with PCV were included in the analysis (8 male, mean age 69 years range 57‐73 years). Mean choroidal thickness was 437 µm ± 97 µm in patients with CSC and mean choroidal thickness was 335 µm ± 53 µm in patients with PCV. The choroid was thicker in PCV and CSC patients than in controls(mean age 62 years range 55‐72, 219 ± 35 µm p<0.005). However, there was significant difference in mean subfoveal choroidal thickness between CSC and PCV (p<0.005) . CONCLUSION: The choroid was thicker in eyes with PCV or CSC than in control
Clinically sensible follow‐up for glaucoma
Authors: LEMIJ H
Published: November 2013

Detecting glaucoma progression is clinically challenging. It is unclear how structural and functional measurements are best used. Data from several long‐term structure and function follow‐up studies into glaucoma progression have revealed a poor overlap between the two, suggesting that one should ideally use both structural and functional measurements for detecting glaucomatous progression in individual cases, with the risk of overcalling and overtreating so‐called progression. With ever more limited resources, however, clinicians are probably forced to restrict their measurements to as few exams as possible in individual patients, calling for sensible choices which measurements to perform. Structural measurements appear to be more useful in early, including preperimetric, glaucoma and functional measurements in more advanced glaucoma. Descriptions, drawings or 2D photography of the optic disc are, on the whole, probably of limited value, whereas standard automated perimetry and imaging of the retinal nerve fibre layer appear to be most useful. This will be discussed in an interactive, clinically oriented presentation. A limitation of this study is that scanning laser tomography of the optic disc has not been evaluated.
Optical measurement of dioptric power and transparency of cornea stored in bioreactor
Authors: PATAIA G , GAUTHIER AS , TRONE MC , CAMPOLMI N , BERNARD A , NAIGEON N , NANGOUM‐FOSSO T , PISELLI S , ACQUART S , LEPINE T , DELBOSC B , THURET G , GAIN P
Published: November 2013

Purpose Quantification of dioptric power (DP) and transparency (T) of stored corneas remain almost unmet in daily eye bank practice. Complementary to T, DP could help screening donors having had refractive surgery and selecting the best donor/recipient refractive matching. During the development of an innovative corneal bioreactor (BR) (patented) for preclinical research and long‐term storage, we took into account the possibility to integrate devices to perform these new quality controls. Aim: to present a device, named “frontofoco‐graft‐meter” (FGM) able to quantify DP and T of corneas in BR

Methods measure of DP was based on the principle of magnification measurement and for T on the transmitted light over incoming light ratio. The FGM comprised a light source + specific chart made of parallel black lines + digital camera. Human corneas in the BR were pressurised at 15mmHg, with medium circulation in endothelial chamber, whereas epithelial chamber was filled with air during measurements. DP was compared to the power map provided by OCT (Tomey, Casia), and T measures to results provided by our previous device called ‘transparometer’ (based on the analysis of modulation transfer function). Measurements were done by 3 operators in a masked fashion

Results For DP, agreement with OCT was good (within ±1 Dioptry). For T, measures were comparable with both devices. Inter‐rater agreement was good for DP and T. FGM took a limited place and could easily be integrate in eye banks

Conclusion the new FGM provides objective and reproducible QC of DP and T of corneas placed in the BR. Grants: ABM 2012, ANSM 2012, EFS 2012
Endothelial and stromal quality control of corneas stored in an innovative bioreactor
Authors: GAUTHIER A , TRONE MC , CAMPOLMI N , NEFZAOUI C , BERNARD A , NAIGEON N , NANGOUM‐FOSSO T , PISELLI S , DELBOSC B , ACQUART S , THURET G , GAIN P
Published: November 2013

Purpose We recently patented a bioreactor (BR) for corneal storage, based on the restoration of a transcorneal pressure gradient and a continuous fluid circulation. Being designed for experimental works and eye banking it comprises 2 windows enabling a right through observation of the cornea. Aim: to assess on human corneas placed in the BR, the possibilities of non‐invasive quality controls (QC) of endothelial cells (EC) and of the stroma (thickness, dioptric power, transparency, detection of scars)

Methods EC QC were: 1/transmitted light microscopy (TLM) with a x10 very LWD objective after trypan blue + osmotic preparation directly performed in the BR; 2/observation with a macroscope (same preparation); 3/specular microscopy (SM, Konan and Hai) without preparation. Stromal QC were: 1/slit lamp; 2/OCT (Heidelberg and Tomey Casia); 3/customized transparometer (analysis of modulation transfer function), and 4/macroscope. Both quality of results and ergonomic aspects were considered

Results EC were visible with TLM and macroscope at highest magnification but not with SM. Casia OCT allowed easy thickness mapping with a good agreement with US pachymetry performed as a control, and dioptric power measurements. Transparency could be controlled with the slit lamp and with our transparometer. The macroscope visualized stromal scars

Conclusion EC and new stromal QC are now possible for corneas stored in the innovative BR. Other new complementary technologies of optical control are currently in development in our lab. Acknowledgments: EFS of Besançon Grant: ABM 2012, EFS 2012, ANSM 2012. PG, GT and SA patented the BR
Congenital hypertrophy of the retinal pigment epithelium (RPE)
Authors: ZOGRAFOS L
Published: November 2013

Congenital hypertrophy of the Retinal Pigment Epithelium (CHRPE) is a tumour which may be confused with pigmented tumours of the choroid such as nevi or melanomas. These lesions are round or oval shaped, with borders that are well defined or scalloped. They present areas of atrophy which may increase in size. This increase in size of CHRPE is well documented and innocent. However, transformation in adenomas or even adenocarcinomas is also possible. In consequence, a periodical observation for Congenital hypertrophy of the Retinal Pigment Epithelium is generally recommended.
Controversies in the surgical treatment of combined hamartomas of the retina and retinal pigment epithelium
Authors: GARCIA‐ARUMI J , VELAZQUEZ VILLORIA D
Published: November 2013

Combined hamartomas of the retina and retinal pigment epithelium are rare benign tumors that may cause significant visual loss. Combined hamartomas are usually solitary, unilateral lesions located at the optic disc or posterior pole. They typically appear slightly elevated and have varying amounts of pigmentation, vascular tortuosity and epiretinal membrane formation. They are usually diagnosed in children or young adults with painless visual loss. Visual function varies with the location of the lesion. Direct involvement of the optic nerve, papillomacular bundle, or fovea may reduce visual acuity. If these structures are not directly involved, visual loss may result from macular distortion by retinal striae and epiretinal membrane. According to the Macula Society’s1 report of 60 patients with combined hamartomas, the tumor was located on the optic disc and adjacent retina in 18% of patients, adjacent to the disc in 28% of patients, extending from the disc to the fovea in 10% of patients, in the macula in 38% and in the mid‐periphery in 5% of patients. Macular location of the tumor is a predictor of poor visual acuity, found in 69% of macular tumors and only 25% of extramacular tumors. Histopathologically, combined hamartomas show evidence of hamartomatous malformation involving hyperplasia of RPE, glial cells, and blood vessels. Epiretinal membranes are an important cause of progressive visual loss, especially in eyes with macular tumors. Indication for epiretinal membrane removal in these patients is not well stablished, and cases reported in literature show different visual results. McDonald et al2 indicated that vitrectomy was not beneficial in their 2 cases, whereas Mason3, Kleiner3 and Stallman4 found benefit in their cases. The surface glial membrane causing the retinal folding is an integral part of the tumor in some cases and accounts for the fact that surgical stripping of the membrane is difficult and has little chance of restoring central vision in such cases5. Surgical intervention should be selected carefully; full‐thickness macular holes have occurred when the membrane is intrinsically woven into the lesion6. It has been speculated that OCT could provide useful information regarding potential visual benefit as the more intact retina on OCT might have better visual outcome than the completely disorganized, thickened retina7. Vitreous traction if present may also be detected, and thus help in selecting cases that may have a potential for visual improvement with vitrectomy and membrane peel to relieve traction.