bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum biomarkers of metastatic disease: current practice and future perspectives
Authors: ANGI M , KALIRAI H , DAMATO B , COUPLAND SE
Published: August 2012

Purpose Molecular genetic testing of primary uveal melanoma (UM) specimens is being integrated into clinical practice for prognostication purposes. However, the current screening protocol for metastatic disease (MD) it is not able to detect micrometastases. There is a clear need for easily assessable biomarkers that would allow earlier detection of MD and aid monitoring of systemic adjuvant therapy. As UM disseminates haematogeneously, it is reasonable to search for blood‐borne biomarkers as indicators of MD.

Methods Review of the literature to provide an overview of blood biomarkers studies in UM. Outline of the workflow, which could lead to new biomarker discovery using a discovery proteomics approach.

Results Conventional biomarkers described in other cancer types have been investigated in blood samples from UM patients. These markers have failed to provide reproducible results and therefore their usefulness in the clinical senario has been questioned. The enumeration of circulating tumour cells shows good potential, but results are influenced by the technique used for detection. Proteomic technology has been applied to UM tumour tissue, secretome and cell lines, mainly using 2D gels. Quantitative mass spectrometry‐based discovery proteomics comparing high risk tumours versus low risk ones, followed by system biology analysis to select a panel of meaningful candidates among the differential expressed proteins, bears the potential to be a more accuare and effective approach.

Conclusion None of the blood biomarkers tested in UM patients have demonstrated the necessary sensitivity and specificity to guide clinical decisions. A rational development of clinically relevant blood biomarkers can be achieved applying proteomic technologies.
Staging of uveal melanoma metastases
Authors: KIVELÄ T , ESKELIN S
Published: August 2012

Purpose To summarize the need and methods available to stage metastatic uveal melanoma.

Methods Literature review and personal experience.

Results In the Tumor, Node, Metastasis (TNM) classification, metastatic disease is assigned to category N1 (regional lymph node metastasis) and M1 (systemic metastasis) and stage IV by definition. For practical purposes, uveal melanoma spawns only systemic metastases. The latest 7th TNM edition, effective from January 2010, introduced subcategories of M1 based on the largest diameter of the largest metastasis (LDLM) at diagnosis of disseminated disease: M1a corresponds to LDLM 3.0 cm or less, M1b to LDLM 3.1‐8.0 cm and M1c to LDLM 8.1 mm or above. In addition to the TNM system, a staging system based on three variables: performance index as a measure of the general health of the patient, serum or plasma alkaline phosphatase level as a measure of the hepatic function and non‐measurable metastases and to LDLM as a measure of measurable metastases is available. It was developed at the Helsinki University Central Hospital and has been validated by the European Ophthalmic Oncology Group. An underlying multivariate model assigns patients into stages IVa, IVb and IVc with predicted median survival of less than 6 months, 6 to 12 months and over 12 months. In the validation data set, the corresponding observed median survival times were 18.6 months for stage IVa, 10.7 months for stage IVb, and 4.6 months for stage IVc.

Conclusion Staging of cancer at diagnosis is important because it enables the physician to assess the extent of disease in a standardized way. This will help in choosing appropriate treatment and in assessing treatment outcomes between studies in a more comparable way.
Therapeutic options in metastatic uveal melanoma
Authors: PIPERNO‐NEUMANN S
Published: August 2012

Purpose Despite the advances in the treatment of uveal melanoma (UM), randomized studies have demonstrated that the metastatic risk was similar after enucleation or conservative treatment. Survival has not improved and enucleation remains the treatment of choice for large tumors.

Methods Up to 50% of patients develop metastases, to the liver in 90% of cases,usually leading to death. Metastases are rarely detected at the time of diagnosis, and in contrast with cutaneous melanoma, occur via hematogenous spread. There is no standard adjuvant treatment to prevent metastases. Today, genome‐wide techniques of genomic and expression profiling make it possible to improve the characterization of high‐risk UM. Recently, mutations in the GNAQ/11 genes have been described as oncogenic drivers in UM and consequently potential therapeutic targets. Since the identification of genomic abnormalities correlated with the metastatic risk, techniques of fine needle aspiration biopsies have been developed and systematic genetic analysis of UM became recommended for the next years, in the prospect of future adjuvant trials based on potential biological targets to be tested in preclinical animal models.

Results Relevant clinical data and dedicated studies in metastatic UM patients are lacking. Only 25 prospective clinical trials were published in the last 30 years,none of them reported a randomized phase III trial. In a retrospective series of 470 metastatic patients managed at Institut Curie (2000‐2008), the median overall survival was 13 months, with a significant difference according to the first treatment in the metastatic setting: 28, 12 and 4 months for liver surgery, any systemic treatment, best supportive care respectively

Conclusion We will review current options and future developments for UM treatment.
Role of corticosteroids in CSCR
Authors: POURNARAS CJ
Published: August 2012

Purpose Central serous chorioretinopathy (CSCR) is a relatively common retinal disease characterized by the accumulation of subretinal fluid at the posterior pole of the fundus, creating a circumscribed area of serous retinal and/or pigment epithelium detachment.

Methods Numerous reports during the past few years have provided us with arguments strengthening the hypothesis of a possible association between CSCR and glucocorticoids.

Results It has been noted that central serous chorioretinopathy is associated with different conditions, characterized by exposure to increased levels of endogenous or exogenous glucocorticoids. In fact, central serous chorioretinopathy has been described in patients with endogenous Cushing’s syndrome. It is also prevalent in patients with type‐A personality; it has been reported following stressful events, whereas pregnancy represents another possible risk factor. These conditions are all characterized by endogenous hypercortisolism. In addition, many cases of central serous chorioretinopathy have been described during or following treatment with glucocorticoids, administrated by any route, for various systemic or ocular conditions. Central serous chorioretinopathy, when related to exposure to exogenous glucocorticoids, has a less prominent male predilection, presents more often with a chronic or atypical form, and is frequently bilateral. Furthermore, treatment of CSCR with glucocorticoids was found to exacerbate the clinical picture. However,very few patients worldwide are suffering from CSCR, while under systemic steroid treatment.

Conclusion It could be suggested that glucocorticoids may be involved in the development of central serous chorioretinopathy. Glucocorticoids should not be used in the treatment of central serous chorioretinopathy.
Photodynamic therapy in the treatment of CSCR
Authors: RUIZ MORENO JM
Published: August 2012

Purpose To evaluate the efficacy of photodynamic therapy (PDT) to treat chronic central serous chorioretinopathy (CSC) and changes in neural retina (NR) thickness and best corrected visual acuity (BCVA) induced by PDT.

Methods Non randomized, multicentric, interventional case series. 82 eyes of 72 patients with chronic CSC were treated by conventional PDT. LogMAR BCVA and central foveal thickness (CFT) measured by optical coherence tomography (OCT) before and after PDT, number of PDT treatments and complications were considered as outcome indicators. In 26 eyes LogMAR BCVA and OCT were evaluated before treatment and one year after PDT. 24 eyes from 24 patients with non chronic, non PDT treated forms of CSC were evaluated as a control group.

Results BCVA changed from 0.53 before PDT to 0.48 at 6 months (p=0.007). Subretinal fluid disappeared in all the cases. CFT decreased from 325 µm, to 229 µm one month after PDT; to 206 µm at three months; and to 202µm at six months (p<0.0001 in the three cases). No cases developed severe visual loss or complications derived from PDT. In subgroup studied at one year NR CFT before PDT was 181.4±42.6 μ vs. 149.0±30.6 μm one year after treatment (p=0.004). NR CFT in the untreated eyes was 204.6±30.7 μ vs. 192.5±26.4 μ after self resolution (p=0.03). Basal NR thickness was not statistically significant different between both groups (P = 0.31); this difference became significant at the end of the follow‐up (P<0.01).

Conclusion PDT may be useful in chronic CSC improving BCVA and reducing subretinal fluid and CFT. The use of PDT in chronic CSC induces NR thickness thinning. This change is not correlated with a decrease in BCVA. Randomized studies with longer follow‐up are needed to asses the real role of this treatment in chronic CSC.
Other therapeutic options
Authors: BEHAR‐COHEN F
Published: August 2012

Abstract not provided
Markers of disease progression in atrophic AMD
Authors: HOLZ F , FLECKENSTEIN M , SCHMITZ‐VALCKENBERG S
Published: August 2012

Geographic Atrophy (GA) is a common cause for severe visual loss in age‐related macular degeneration (AMD). In contrast to neovascular AMD in which efficient therapy is now available with intravitreally administered inhibitors of VEGF, there is as yet no therapy available for patients with GA. Natural history studies have demonstrated a continuous progression of outer retinal atrophy in patients with GA. Hereby, a high interindividual variability has been documented with regard to enlargement rates of GA ptaches over time. Various markers of disease progression have been identified including phenotypic charatersitcs on fundus photographs such as soft drusen and hyperpigmentary changes. Furthermore, certain abnormal patterns of increased fundus autofluorescence (FAF) on confocal SLO images have been shown to represent risk characteristics for GA evolution and aloow disrimintaion of slow vs. fast progressors. The status of the fellow also has an imapct on progression rates. SD‐OCT imaging is currently evaluated in ongoing longitudinal studies with respect to specific microstructural alterations in the junctional zone of GA and their impactz for future progression. Idenfication of progrnostic markers is not o9nly important for a better understandibng of the disease process, but also for the design and conduct of interventional clinical trials in patients with GA as enrichment for fast progressors allows to perform such trials in an acceptable time period in a relatively slowly progressing disease, to reduce costs for clinical developments and to increase the likelyhood for the identification of a therapeutic signal.
Gene therapy in rare diseases
Authors: SAHEL JA
Published: August 2012

Purpose Stargardt disease is caused by a mutation of the ABCR gene leading to photoreceptor degeneration and vision loss. Genetic replacement StarGen™ uses LentiVector® technology to deliver a corrected version of the ABCR gene. A single administration of the product directly to the retina could provide long‐term or potentially permanent correction. The primary objective of StarGen™ study is to assess the safety and tolerability of ascending doses of StarGen™ in adult patients with Stargardt macular degeneration. Up to 28 patients 18 years or older with a differing level of advancement of SMD will be included in this study in which a dose‐escalation phase of StarGen™ is followed by a dose confirmation phase. In Retinitis Pigmentosa, preservation or partial restoration of impaired cone function offers very promising perspectives based on neuroprotection, prosthetics or optogenetics. We demonstrated that cone cell function loss might result from the loss of expression of Rod‐derived Cone Viability Factor (RdCVF) consecutive to degeneration of rod photoreceptors directly affected by causative mutations. Administration of RdCVF, irrespective of the gene defect, induced in relevant animal models a strong preservation of cone cell function related to the maintenance of rod outer segments (1). Recently, work conducted by Botond Roska and our group showed that in advanced cases, cone cell bodies of dormant cones can be reactivated by vectorization of halorhodopsins (2). Over the past years we have followed a cohort of over 3000 patients and studied the morphology and function of cone photoreceptors, using novel high resolution imaging.. We showed that cone outer segment degeneration during the disease course can be documented and that patients suitable for clinical trials testing neuroprotection or optogenetics can be selected on this basis. A comprehensive personalized set of therapeutic strategies can be tailored on this basis. Such advances, unprecedented in the field of currently untreatable retinal diseases, offer hope for visual rehabilitation. To evaluate and document such functional outcomes, we have developed novel tools for assessing reproducibly visual impairment and restoration, as well as palliative aids and associated training protocols.

Conclusion StarGen™: A Phase I/IIa Dose Escalation Safety Study of Subretinally Injected StarGen™ Administered to Patients With Stargardt Macular Degeneration Sponsored by Oxford BioMedica UK Ltd. Principal Investigator Prof. Jose‐Alain Sahel Centre Hospitalier National d’Ophtalmologie des Quinze‐Vingts, Paris France – Professor David Wilson ‐Casey Eye Institute, Portland Oregon Léveillard T, Sahel JA. Rod‐derived cone viability factor for treating blinding diseases: from clinic to redox signaling. Sci Transl Med. 2010 Apr 7;2(26):26ps16. Busskamp V et al. Genetic reactivation of cone photoreceptors restores visual responses in retinitis pigmentosa. Science. 2010 Jul 23;329(5990):413‐7.
Perspectives for new treatments at Novartis
Authors: BURIAN G
Published: August 2012

Clinical research in the current clinical practice and regulatory environment related to development of Medicinal Products has evolved in recent years to encompass areas beyond the primary aim of achieving a Marketing Authorization. A model of “development continuum” appears to recently emerge to address key aspects in this new setting: the area of personalized medicine and specialty care, and associated outcomes research, which together aim to achieve therapeutic optimization at individual patient level, in both endemic as well as in rare diseases. This type of clinical research is conducted in late development phase of drugs, most often post‐marketing, under continuous regulatory guidance for drug label optimization. Novartis recognizes and is interested in this changing environment, and is hence committed to address these emerging unmet medical needs that impact clinical practice. Collaboration with the wider group of healthcare professionals in the global community to facilitate the design and implementation of clinically relevant research projects is an important aspect to drive scientific and medical progress in this area. In this presentation, concepts of clinical development and trials design for optimization of drug outcomes and therapeutic patient management will be discussed from an industry perspective. The regulatory and Novartis guidelines for supporting them will be explained, using focussed examples from the field of Ophthalmology, particularly areas of research interest in chorioretinal vascular diseases.
Study Assessing Double‐masKed Uveitis tReAtment (SAKURA)
Authors: NAOR J
Published: August 2012

Purpose The purpose of this study is to evaluate the safety and efficacy of intravitreal injections of DE‐109 ophthalmic suspension.

Results Primary Outcome Measures: Proportion of subjects with vitreous haze score [Time Frame: 5 months] [Designated as safety issue: Yes]

Conclusion Study Design: Allocation: Randomized Endpoint Classification: Safety/Efficacy Study Intervention Model: Parallel Assignment Masking: Double Blind (Subject, Caregiver, Investigator, Outcomes Assessor) Primary Purpose: Treatment
Parainflammation and the ageing retina
Authors: XU H
Published: August 2012

Purpose To review para‐inflammatory responses in the ageing retina, and the role of dysregulated para‐inflammation in age‐related retinal degeneration.

Methods Published and unpublished data from personal study in in vivo models and literature review.

Results Para‐inflammation is an immune response to chronic noxious stimuli at a low magnitude that lies between the basal homeostatic state and overt inflammation. The physiological role is to maintain tissue homeostasis and functionality. The retina, particularly the macula, is constantly subjected to a low‐level of insult mediated by oxidized molecules such as unsaturated fatty acid, and the insult accumulates with age. Retinal homeostasis in the ageing eye is maintained by a para‐inflammatory response executed by resident microglia, retinal pigment epithelial (RPE) cells and the complement system. A dysregulated para‐inflammation may not be able to maintain retinal homeostasis, and age‐related retinal degeneration may occur. The presentation will discuss how microglial activation is controlled in the retina, and the potential contribution of uncontrolled microglial activation to age‐related retinal degeneration.

Conclusion Para‐inflammation is important to retinal homeostasis in the ageing eye. Uncontrolled or dysregulated para‐inflammation may contribute to the pathogenesis of age‐related retinal degeneration.
Detecting progression using structural measurements
Authors: BRON A
Published: August 2012

The management of glaucoma is driven by the changes of functional and structural endpoints overtime. Is the disease stable or not? In this presentation we will cover the practical aspect of structural progression assessment which follows the same rules that apply to functional tests; good baseline material and sufficient further examinations whatever the technique. We will detail the methods used to assess structural progression which are similar to those used for evaluating functional progression. Although challenging, the evaluation of structural progression is very helpful in the follow‐up of the glaucomas. However it must be combined with functional progression to achieve clinical relevance.
Into practice: clinical examples
Authors: STALMANS I
Published: August 2012

Clinical examples on the use of visual field and structural measurements in the management of glaucoma patients will be presented, and discussed with the audience.
GCA diagnosis controversies
Authors: HAWLINA M
Published: August 2012

Purpose To report on different clinical presentations of giant cell arteritis (GCA, temporal arteritis) and diagnostic and therapeutic approach. GCA is the most common primary vasculitis of adulthood. Ocular symptoms present in 30‐75% of patients and include visual loss, amaurosis fugax, ophthalmoplegia, diplopia and ocular pain. Approximately 15‐50 % of patients suffer from permanent visual loss. Temporal artery biopsy and corticosteroid treatment is essential for diagnosis and preservation of vision.

Methods Review and retrospective study of 50 eyes (36 patients) with giant cell arteritis treated at Eye Hospital Ljubljana. There were 67% female and 33% male patients with average age of 79 years, range 62‐92 years, with typical clinical picture and/or positive biopsy (78%, 18/23 performed).

Results Loss of vision was unilateral (65%) or bilateral (35%). The most common ischaemic lesions were anterior ischaemic optic neuropathy (AION; 16 unilateral, 4 bilateral) and posterior ischaemic optic neuropathy (PION; 5 unilateral, 5 bilateral). Other lesions included central retinal artery occlusion and internuclear ophthalmoplegia. More than half of patients (53%) had visual acuity of counting fingers or less in affected eye. In AION various peripheral defects almost universally showed respect for horizontal meridian. Nasal and inferior halves of visual fields were most often affected. PION most often presented with scotomas with or without peripheral defect.

Conclusion Severe loss of vision is present in affected eye in most patients. Bilateral loss of vision can occur if the diagnosis is not established early and corticosteroids given. Very few affected eyes recover to small extent. It is essential to recognise this disease even in absence of typical signs and symptoms. In elderly, all the patients to report amaurosis fugax, sudden loss of vision or diplopia should be suspected to have temporal arteritis.
Atropine use in childhood myopia treatment
Authors: GRZYBOWSKI A
Published: August 2012

Purpose To discuss the role of atropine use in childhood myopia treatment.

Methods The analysis of the literature and studies of the subject.

Results The prevelance of myopia the population in the United States, Europe and Australia is about 30‐40%, whreaes in some southern Asian countries is up to 80‐90%. Moreover, it was shown that this prevalence has been increasing in the recent 20 years. Although the pathogenesis of myopia is unknown, it is argued that, at least in the case of school myopia, environmental factors are of major imprtance, whrease genetic factors play secondary role. The role of atropine in the lowering the rate of myopia progression was analysed in recent 10 years in many studies, differing with the atropine concentration, studied group characteristics, the time of treatment and the the time of observation, etc. The lowering effect of atropine was proved, however, its effectiveness decreased with the time of drug use, and after the drug discontuinuation the effect was partially reversed. On the other hand, it is well‐known that atropine use is related to significant side‐effects, including mydriasis, cycloplegia, increased exposure to UV light. Moreover, some argue that results of the therapy, i.e. lowering of the magnitude of school myopia, cannot be be balanced by the problems and unknown longterm effects related to the therapy.

Conclusion The atropine use in childhood myopia treatment remains controversial and possible benefits and weaknesses of the therapy should be carefully analysed.
Is the eye a window to the mind? RNFL thickness measurements by OCT as a biomarker for neurodegenerative diseases
Authors: ASCASO F
Published: August 2012

Purpose Optical coherence tomography (OCT), a noninvasive imaging technique that assesses the retinal nerve fiber layer (RNFL) thickness, macular thickness and volume, is widely used in various ophthalmologic disorders including glaucoma and macular diseases. A decreased RNFL thickness can correspond to neuronal death and axonal loss. Therefore, OCT measurements might be of particular interest in numerous neurodegenerative diseases with axonal loss.

Methods A significant reduction in the peripapillary RNFL thickness has been reported in patients with various neurologic diseases, such as multiple sclerosis, neuromyelitis optica, Alzheimer´s disease, and Parkinson´s disease, suggesting that OCT might also prove useful in other neurodegenerative disorders. Thus, we have studied for the first time these neurodegenerative changes in patients with schizophrenia and obstructive sleep apnea syndrome (OSAS). The goals of these studies were to determine by OCT the differences in the peripapillary RNFL thickness, macular thickness and volume, between controls and patients with several neurodegenerative disorders in which there is axonal loss, and to assess whether a correlation exists between the RNFL thickness and the clinical severity of the disease.

Results Most neurodegenerative disorders show a significant thinning of the peripapillary RNFL; however, the patterns of change differ in some aspects. The findings indicate loss of retinal ganglion cells and may reflect degenerative brain changes in these conditions. This topic reviews the application of OCT imaging of the retina in Neurology and Psychiatry.

Conclusion RNFL thickness may be used as a biological marker and might help to the early diagnosis of several neurodegenerative conditions.
The influence of peripheral refractive errors on myopia development
Authors: OHLENDORF A
Published: August 2012

Purpose This talk will present research about the role of peripheral refraction on the development of central refractive errors in human eyes as well as in various animal models of myopia.

Methods The interconnection of peripheral refractive profiles and central refractive error will be described. Studies investigating the role of the peripheral refractive error on central refractive development will be presented.

Results Myopes typically show relative hyperopia in the periphery of the eye, while emmetropes and hyperopes show a relative peripheral emmetropia or myopia. Studies in humans as well as in various animal models of myopia indicate that the management of the peripheral refraction can change the development of central myopia. More recently, Smith et al. confirmed the influence of peripheral refraction on myopia development in monkeys, by influencing peripheral refraction with either form‐deprivation (2005) or minus lenses (2009).

Conclusion Research in animal models of myopia and in humans found strong evidence that the peripheral refraction of the human eye can guide the development of central refractive errors. More studies are needed investigating the efficiency of “peripheral refraction management” with various kinds of correction methods (e.g. Contact lenses, Ortho‐K, spectacle lenses). Additionally, there is a need for long term studies on central refractive development based on peripheral refraction in the human eye.

Commercial interest
Proteomic analysis of uveal melanoma
Authors: RAMASAMY P , HENRY M , LINGE A , HORGAN N , MURPHY C , CLYNES M , KENNEDY S , MELEADY P
Published: August 2012

Purpose To further the molecular biological understanding of the events governing the development of metastatic disease and to identify potential targets for therapy.

Methods 8 metastatic vs 8 non‐metastatic fresh uveal melanoma tissue with a minimum follow‐up of 10 years were subjected to quantitative, label‐free proteomic analysis. Strict criteria was applied in the Progenesis LC‐MS software to filter out statistically non‐significant peptides and proteins. These were anova p < 0.05, power > 0.8 and protein confidence score > 100. Differential expression of proteins and their relative abundance in the 2 disease groups were determined using the same software.

Results 653 proteins were identified (global proteome of all 16 samples). Of these, 61 proteins met the statistics criteria applied. 9 of those had good separation between metastatic and non‐metastatic disease groups. 4 proteins were selected for further follow‐up and validation. Currently, immunohistochemistry studies are being performed to confirm expression levels in paraffin embedded tissue slides. Then, siRNA knockdown and immunoprecipitation experiments will be performed in uveal melanoma cell lines (primary and metastatic) to determine downstream effects and potential targets for therapy.

Conclusion Proteomic analysis of 8 metastatic vs 8 non‐metastatic fresh uveal melanoma tissue has identified 4 proteins which are currently being validated using immunohistochemistry, siRNA knockdown and immunoprecipitation experiments.
Temperature‐dependent vascular endothelial growth factor (VEGF) induction in human retinal pigment epithelium – implications for transpupillary thermotherapy in uveal melanoma
Authors: KLETTNER A , FABY H , HILLENKAMP J , ROIDER J
Published: August 2012

Purpose Transpupillary thermotherapy is a hyperthermia treatment for small uveal melanomas but its use is controversially discussed. In uveal melanomas, vascular endothelial growth factor (VEGF) increase is correlated with metastases, and VEGF increase can be found in treated melanomas. In this study, the effect of hyperthermia on the VEGF secretion in human RPE cells was studied.

Methods Immortal human retinal pigment epithelium (RPE) cell line Arpe‐19 was exposed to 40°, 42°, 45° and 50°C for 1 min, 5 min and 15 min. Toxicity was evaluated using trypan blue exclusion assay and VEGF secretion was evaluated by ELISA. Involvement of Mitogen activated protein kinase (MAPK) pathways and Transient Receptor Potential Vanilloid (TRPV) channels on VEGF induction was investigated using commercially available inhibitors.

Results Hyperthermia induces cell death in a time and temperature dependent manner. VEGF expression and secretion is strongly induced by hyperthermia in a time and temperature dependent manner. VEGF induction is mediated by p38 and to a lesser degree by JNK. TRPV channels are only involved in VEGF secretion at lower temperatures.

Conclusion Hyperthermia induces a temperature dependent secretion of VEGF in human RPE, which is mediated by p38. As VEGF may be involved in the development of micrometastases, these findings indicate that thermotherapy for the treatment of uveal melanomas should be regarded with caution.
Ultrasound features changing during uveal melanoma local treatment
Authors: AMIRYAN AG , SAAKYAN S
Published: August 2012

Purpose The aim of our study was to estimate the echographic sighs (acouctic densitometry characteristics and hemodynamic changes in tumoral vascular network) of uveal melanoma during the local irradiation.

Methods 58 patients (31‐74 years old) with uveal melanoma underwent the investigation. Acoustic densitometry and hemodynamic characteristics in tumoral vessels were estimated both before and during brachytherapy using high‐resolution duplex scanning (using ultrasound system Voluson® 730Pro, GE Medical Systems, Kretz Ultrasound).

Results There were estimated some alterations in echographic sighs which included the increasing of blood flow velocity in tumoral vessels and decreasing the acoustic densitometry characteristics during the local treatment in comparison with those before the treatment. The dimension and the degree of intensity those alterations depended on tumor size, it’s localization in the eye, facility of the irradiation, angioarchitecture of the lesion.

Conclusion The estimation of echographic alterations in uveal melanoma during the local treatment allows indirectly to judge about tumor radiosensivity and probably to assume the local treatment efficiency.
Risk factors for endothelial cell loss after corneal transplantation
Authors: BORDERIE V , BULLET J , TOUZEAU O , BASLI E , LAROCHE L
Published: August 2012

Purpose To analyze risk factors for endothelial cell loss after corneal transplantation.

Methods Cohort study including 1438 consecutive eyes of 1438 patients who underwent Penetrating Keratoplasty, Anterior Lamellar Keratoplasty, or Descemet Stripping with Endothelial Keratoplasty between 1992 and 2010. To describe post‐operative endothelial cell loss, the biphasic linear model was fitted to grafts with at least 2 (early phase) or 3 (late phase) endothelial cell density (ECD) measurements in time: ECD = ECD0 – (t * E), for t < 1 year (early phase); ECD = A – (t * L), for t > 1 year (late phase). The least squares estimates were used to calculate the coefficients of the various equations for each eye.

Results The average follow‐up time was 45.9+39.3 months (mean+SD). The early‐phase and late‐phase annual endothelial cell loss (ECL) were, respectively, ‐24.7+18.8% (n=921) and ‐10.8+10.3% (n=664). Overall, the model‐predicted cell loss was lower than the observed cell loss in the early phase and higher in the late phase. In multivariate analysis, 5 variables significantly influenced the early‐phase endothelial cell loss: graft storage time, recipient rejection risk, preoperative hypertony, surgical group, and rejection episodes. Three variables significantly influenced the late‐phase endothelial cell loss: recipient rejection risk, surgical group, and rejection episodes.

Conclusion Extending the graft organ culture time beyond 3 weeks results in higher early cell loss. High‐risk recipient and rejection episodes are important risk factors for cell loss. ALK induces minimal cell loss whereas keratoplasty in eyes with impaired recipient endothelium is associated with high cell loss.
Delivery of molecules into corneal endothelium using nanoparticles activated by femtosecond laser pulses: proof of concept
Authors: JUMELLE C , CAMPOLMI N , BERNARD A , PISELLI S , AUDOUARD E , GRANIER J , SODER H , GAIN P , THURET G
Published: August 2012

Purpose NanoFemtoTransfection (NFT) is an innovative and promising non‐viral technique to transfer molecules into cells (Chakravarty. Nature Nanotechnology 2010;5:607). It consists in temporarily permeabilizing cell membrane by a photoacoustic effect obtained by nanoparticles of black carbon activated by Ti‐Saphir femtosecond laser (fsL) pulses. Calcein (622 Da), tagged bovine serum albumine (70 kDa) and one eGFP plasmid (5 MDa) were transfected into two non‐adherent cell lines (DU145 prostate‐cancer and GS‐9L rat gliosarcoma). Our aim was to adapt the NFT to adherent human corneal endothelial cells (HCEC)

Methods We tested the NFT of calcein in vitro on the HCEC line HCEC‐12 seeded at 1500cells/mm2 in 6 wells plates and ex vivo on whole human organ cultured corneas. A matrix of experiments comprising 4 exposition times, 6 fluences and 2 fsL beam movements was performed in order to obtain transfection with minimal toxicity. After exposition to fsL, nuclei were counterstained with Hoechst33342 and transfection efficiency was determined by observation on a fluorescence inverted microscope (IX81, Olympus, Japan) and further quantified by flow cytometry (FACSCalibur, BD, CA). Viability was assessed by Trypan blue staining

Results In HCEC‐12, a fluence of 100 mJ/cm² and a laser beam movement of 3,5 mm/s gave a transfection of 17% and a viability of 97%. In whole corneas, with the same parameters, transfection was detectable in disseminated EC

Conclusion We obtained the proof of concept of the NFT in HCEC. Further optimization is ongoing to increase the transfection rate while maintaining minimal toxicity, especially for bigger molecules, like plasmids
Characterization of cell cycle modifications induced by electric pulses in human corneal endothelium
Authors: HA THI BM , HE Z , CAMPOLMI N , PISELLI S , GAIN P , PEOC'H M , DUMOLLARD JM , ACQUART S , GARRAUD O , THURET G
Published: August 2012

Purpose We recently described the possibility of gene electrotransfer into endothelial cells (EC) of human organ cultured corneas (He. OphthalmicRes2010;43:43). Parallel to the transfection of plasmids, trains of electric pulses alone also triggered mitosis of EC, despite human endothelium is normally considered unable to proliferate. Our aim was to further characterize the effect of electric pulses on the cell cycle of EC

Methods The short‐term (3 days) and long‐term (30 days) effects on EC of 8 square pulses lasting 100 ms, at 1 Hz frequency and 125 mA intensity were studied on 20 human organ cultured corneas. EC proliferation was evaluated by quantification of Ki67 expression, a proliferation marker of G1 to M phase, and by 5‐Ethynyl‐2’‐Deoxyuridine (Clik‐it EdU) incorporation during DNA replication (S phase marker) EC viability was assessed using the triple labeling with Hoechst33342/Ethidium/Calcein‐AM with calculation of the viable EC density by image analysis (Pipparelli.IOVS2011;52:6018) and using an apoptotic marker, pSIVA‐IANBD

Results Three days after stimulation by electric pulses, a significant activation of EC proliferation was observed with S‐phase revealed by EdU staining and G1 to M phase, including mitosis, revealed by Ki67 staining. 90% of proliferating EC were located in area with ECD bellow 2000cells/mm2. For long‐term effect, ECD did not increase and nuclear fragmentation and cell death were detected

Conclusion Electric pulses activate the cell cycle of EC on ex vivo corneas but cell division seems to abort and result in cell death. Mechanisms of cell cycle trigger and of mitosis abortion are currently being studied.
Impact of corneal donor post‐mortem time on the rate of culture medium contaminations
Authors: GRUENERT A , ROSENBAUM K , GEERLING G , FUCHSLUGER T
Published: August 2012

Purpose In the majority of European countries there is a shortage of donor corneas for corneal transplantations. In 2007, the EU Directive 2006/17/EC on ‘technical requirements for the donation, procurement and testing of human tissues and cells’ was introduced restricting the usability of post‐mortem donor blood sampling from 72 hours to 24 hours. As a consequence, post‐mortem time for donor cornea acquisition was shortened, resulting in a further reduction of corneal allografts. This study analyzed the effect of donor cornea post mortem‐time on medium contamination in organ culture storage.

Methods Contamination rates during corneal organ culture during extended (2008‐9, Group I) versus restricted post‐mortem time protocols (2010‐11, Group II) were retrospectively analyzed. Data were collected at LIONS Cornea Bank North Rhine‐Westphalia, University Eye Hospital Duesseldorf, Germany.

Results In the years 2008 and 2009 (Group I) 1272 corneal grafts with a post‐mortem time of 30.05 ± 15.77 hours were collected and cultivated in LIONS Cornea Bank NRW. After introduction of the new guidelines, the overall number of acquired donor corneas dropped by 29.88% to 892. Post‐mortem time was significantly lower during the restricted post‐mortem time protocols (24.2 ± 12.37 hours, p<0.05, t‐test). Interestingly, there was no significant difference of culture medium contamination rates between the two groups (Group I: 9,0%, n=115 versus Group II: 7.3%, n=65; Chi‐squared test).

Conclusion Restriction of donor cornea acquisition time down to 24 hours seems to be without benefit regarding the contamination rates during corneal organ culture.
Big bubble technique dissection plane: histological and ultrastructural comparative analysis on both white and clear margin dissected corneas
Authors: IRION L , MCKEE H , BRAHMA A , CARLEY F
Published: August 2012

Purpose Pneumodissection or 'big‐bubble' technique (BBT) is used to separate Descemet’s membrane (DM) and stroma in both deep anterior lamellar keratoplasty and Descemet’s membrane endothelial keratoplasty. It has been reported that BBT may produce two typer of bubble outlines: clear or white margin. It seems that the components of the posterior lamella obtained with each bubble rims also differ. Our aim is to compare BBT dissection plane using histological and electron microscopy (EM) analysis on eye‐bank corneas.

Methods BBT was carried out on donor corneas unsuitable for transplantation for reasons other than corneal disease.Corneoscleral discs were mounted on an artificial anterior chamber and BBT was carried out using a 27G needle. 5 samples which had a bubble with white margin and 5 with clear margin were fixed in formalin and sent for histology followed by EM.

Results Samples in which the edge of the bubble had a white margin showed variable residual stroma adherent to DM. When the edge of the bubble was clear no residual stroma was seen attached to a bare, smooth DM. The anterior lamellae of the cases without residual stroma were also analysed. These showed no evidence of DM elements excluding the unlikely possibility of intra‐Descemet’s dissection plane. The findings were confirmed by EM in all cases. These showed intact banded DM in all posterior lamellae but only those with a white bubble outline had attached residual stromal collagen.

Conclusion In eye bank corneas the histological elements of the posterior lamella dissected using BBT can be predicted depending on the margin of the bubble obtained. Assumptions that pneumodissection in BBT routinely bares DM need to be revisited.
Clinical challenges of ocular anti‐microbial therapy
Authors: RAJU L
Published: August 2012

Purpose To discuss the different clinical challenges in ocular anti‐microbial therapy as they pertain to bacteria, fungi, viruses, and acanthamoeba.

Methods An overview of how the different infections often seen and how available medications might be utilized.

Results With the large number of medications available to ophthalmologists these days, deciding what medication to use can be complicated. The initial choice of fourth‐generation fluoroquinolones has raised questions about bacterial resistance. This may highlight the importance of cultures in cases where infections are not responding as expected. Recognition of certain infections, such as early acanthamoeba, which can look similar to herpetic keratitis, may be difficult. The use of newer diagnostic tests, such as PCR, may be helpful to ensure appropriate treatment is started in a timely fashion. This may also be helpful in targeting treatment in fungal infections. The use of long term wear contact lenses, such as with a keratoprosthesis, has raised questions as to whether antibiotics are efficiently reaching the corneal surface and treating biofilms that form on contact lenses. Understanding what flora is present in cases of long term contact use may allow clinicians to better prevent infections. There have also been many clinicians who have advocated the treatment of viral conjunctivitis to help reduce the number of lost work and school hours. There is still debate as to the efficacy of these treatments and how it might impact the continually growing cost of health care.

Conclusion Newer testing modalities and understanding what long term use of ocular anti‐microbials does on the ocular surface will improve our treatment of ocular infections and hopefully prevent them too.
Is antibiotic resistance a major problem in ophthalmology?
Authors: KOWALSKI R
Published: August 2012

Purpose Ocular antibiotic bacterial resistance becomes problematic when: 1) intrinsically susceptible bacteria become resistance, 2) antibiotic resistance spreads from patient –to‐patient, and 3) effective antibiotic treatment is unavailable. The “Standard of Care” of ocular bacterial infections will be accordingly evaluated.

Methods Based on the findings of our dedicated ophthalmic microbiology laboratory, the trends in ocular susceptibility in regards to bacterial pathogen and disease (endophthalmitis, keratitis, conjunctivitis, and blepharitis) were analyzed for evidence of acquired resistance, possible infection spread, and the armament of treatment.

Results Our studies indicate that based on the bacterial pathogen, bacterial resistance is most prevalent with endophthalmitis isolates of Staphylococcus aureus and coagulase‐negative Staphylococcus. There is evidence that endophthalmitis isolates of Staphylococcus aureus may be hospital acquired. Although Haemophilus influenza and Streptococcus pneumonia isolated from conjunctivitis are probably spread patient‐to patient, an increase in antibiotic resistance has not occurred. There does not appear to be a treatment problem for Pseudomonas aeruginosa keratitis based on in vitro antibiotic resistance.

Conclusion Our study concludes that antibiotic resistance does occur for ocular bacterial pathogens, but ocular infection does not spread from patient to patient. There is a plethora of effective topical antibiotic agents to treat all types of bacterial ocular infections. It is important to culture all severe ocular infections to optimize therapy and to guard against the creation of antibiotic resistant bacteria. The future of antibacterial therapy may rely on preventing infections instead of the production of new drugs.
Historical concepts of glaucoma
Authors: GRZYBOWSKI A
Published: August 2012

Purpose To present different historical concepts of glaucoma in relation to the thought‐styles of their time.

Methods Historical analysis of the old medical treatises and modern history of ophthalmology articles.

Results The pathogenesis of glaucoma is one of the most important and puzzling issues in the 21st century ophthalmology. Many concepts were recently presented, including light‐damage theory, local and systemic ischaemic injury, oestrogen deficiency and low CSF pressure. It shows its multifactorial profile and our present deficiency in understanding its real nature. The possible reason for this might be our scientific thought‐style, which determines our present perception of the disease and possible concepts of its pathogenesis, which are verified in clinical and experimental research. The collective thought‐style, in this case regarding the nature of glaucoma, might easily eliminate any new idea or non‐conventional hypothesis. The scientist is a part of the thought‐collective. Every observer has been shaped by a particular culture and represents a thought‐style of a definite scientific group, and his style determines the range of his observations. The historical glaucoma concepts reflected usually historical thought‐styles typical for the period of scientific development.

Conclusion Understanding the principles that govern the development of science in the historical perspective allows for an understanding of our own limitations in this respect (contemporary thought‐style), which frequently prevent us from looking at the matter in a non‐standard way.
Pathogenesis of glaucoma: does light play a part in the process?
Authors: OSBORNE N
Published: August 2012

Pathogenesis of glaucoma: does light play a part in the process? By: Neville N. Osborne Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernández‐Vega, Oviedo 33012, Spain. Neurones are absolutely dependent on their mitochondria for their energy needs unlike dividing non‐neuronal glial cells. Retinal ganglion neurones are particularly dependent on their mitochondria because a lot of energy is required to propagate the action potential along the non‐mylinated axons within the globe. We have propose that glaucoma is initiated by an alteration in the blood flow dynamics in the optic nerve head causing a defined “ischemic‐like” insult to the retinal ganglion neurone axon mitochondria and also surrounding astrocytes and microglia. While this results in ganglion cells being metabolically weaker, but still functionally normally, they are now susceptible to secondary insults that they would have normally tolerated. The major secondary insults to subsequently result in ganglion cells dying at different times probably arise from chemicals released from activated optic nerve head astrocytes and microglia. Yet another factor may be the very same light that is involved in vision. This is because basic science studies have shown that light as impinging on ganglion cell axons can affect mitochondrial function. Thus while it is not suggested that light causes glaucoma it is proposed that particularly the blue light component impinging on the retina can exacerbate already metabolically effected ganglion cell mitochondria.
Vascular tumours of the retina and choroid : classification, diagnosis and treatment
Authors: SCHALENBOURG A
Published: August 2012

Purpose Retinal vascular tumors consist of capillary hemangioblastomas, associated with Von Hippel Lindau’s disease, cavernous or racemous hemangiomas, and vasoproliferative pseudotumors. Coat’s disease is typically diagnosed in male children presenting retinal telangiectasia, complicated by retinal exudates. Choroidal vascular tumors are most frequently circumscribed, more rarely diffuse hemangiomas, the latter being associated with Sturge Weber syndrome

Methods Diagnosis is often made by ophthalmoscopy and ultrasonography, and confirmed by fluorescein and/or ICG angiography. OCT and panoramic angiography have improved visualization with regard to the extent of the vascular tumor(s).

Results Therapeutic approach can consist of periodic observation, laser photocoagulation, cryotherapy, photodynamic therapy (PDT), irradiation by plaque or protons, and anti‐VEGF injections.

Conclusion Controversy exists as to the optimal management of these benign but potentially blinding tumors. This review therefore serves as an introduction to the different topics of this symposium as well as the debate at the end with different experts sharing their experience.