Time table

Program

Final Program

 

Download the ISOPT Clinical 2016 Final Program

Timetable 2015

ISOPT clinical 2015 timetable

*Updated 1 July, 2015 

Speakers 2015

M. Ader, Germany
E. Akpek, USA
S. Androudi, Greece
P. Asbell, USA
P. Ashton, USA
A. Augustin, Germany
A. Auricchio, Italy
W. Ayliffe, UK
F. Bandello, Italy
I. S. Barequet, Israel
T. Barisani, Austria
N. Barney, USA
A. Bate, USA
F. Behar-Cohen, Switzerland
R. Beuerman, Singapur
F. Boscia, Italy
J. Brandt, USA
K. Brazzel, USA
M. Bolz, Austria
S. R. Boyd, Canada
J. D. Bullock, USA
J. Busik, USA
B. Butler, USA
A. Chauhan, USA
S P. Chee, Singapore
R. Chuck, USA
J. Ciolino, USA
G. Coscas, France
D. Dalkara, France
A. Denniston, UK
P. Dugel, USA
Y. El Shabrawi, Austria
I. El-Zaoui, France
M. Eydelman, USA
R. Feldman, USA
B. Flowers, USA
L. Fontana, Italy
C. Francois, USA
F. Fraunfelder, USA
A. Garcia Layana, Spain
D. Gabriel, Switzerland
D. Ghezzi, Switzerland
J. Goldberg, USA
N. Goldenberg -Cohen
M. Goldstein, Israel
M. Grant, USA
J. Groot-Mijnes, The Netherlands
Y. Guex-Crosier, Switzerland
F. Grus, Germany
A. Grzybowsk, Poland
P. Hamrah, USA
P. Harasymowycz, Canada
A. Harris, USA
R.V. Herrero, Spain
D. Heuer, USA
D. Hinton, USA
J. Reidy, USA
B. Jeng, USA
J. Jonas, Germany
M. Kasper, Germany
M. S.J. Katz, USA
B. Katz, USA
H. Kaufman, USA
C. Kostic, Switzerland
U. B. Kompella, USA
R. P. Kowalski, USA
R. Kowluru, USA
C. Kostic, Switzerland
M. Kramer, Israel
B. D. Kuppermann, USA
H. Leiba, Israel
A.Leonardi, Italy


T. Leveillard, France
L. A. Levin, Canada
B. Levy, USA
W. H. Lee, USA
A. Ljubimov, USA
N. Lois, Ireland
B. Lumbroso, Italy
W. MacVicar, USA
I. Mantel, France
A. Matet, Switzerland
N. Medow, USA
O. Moghimi, USA
S. Mohr, USA
R. Neumann, Israel
G. Noronha, USA
G. D. Novack, USA
F. Obermayr, Austria
A. Ohira, Japan
E. Ongini, USA
M. Paques, France
C. Parsa, USA
S. Patel, USA
J. Penn, USA
L.E. Pillunat, Germany
C.S. Phaik, Singapur
U. Pleyer, Germany
T. R Willis, USA
M. J. Rafii,USA
CA Rasmussen, USA
M. R. Rodriguez, Switzerland
H. Reitsamer, Austria
P. Roberts, Austria
E. Romanowski, USA
B. Roska, Switzerland
SV R. Sadda, USA
J.A. Sahel, France
V. Santer, Switzerland
P. Sieving, USA
L. Schmetterer , Austria
S. Schmitz-Valckenberg, Austria
J. M. Seddon, USA
M. Seiler, USA
R. M Shanks, USA
K. Shindler, USA
J. Shultz, USA
D. Silverman, USA
K. Singh, USA
J. Slakter, USA
A. Sodhi, USA
A. Solomon, Israel
G. Soubrane, France
S. Sziliak, USA
G. L. Spaeth, USA
J. Sparrow, USA
O. Stachs, Germany
B. V. Stanzel, Germany
G. Staurenghi, Italy
E. Stefánsson, Iceland
C. Struble, USA
N. Stübiger, Germany
F. Thieltges, Germany
M. Thormann, USA
E. Timm, USA
A. Timmers, USA
R. Toyos, USA
O. Ucakhan-Gunduz, USA
P. Udaondo, Spain
S. Uretsky, France
M. Vezina, Canada
N. Waheed, USA
K. Weber, Switzerland
M. Zierhut, Germany

 

 

 

Free Papers Abstract 2015

942

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Retina

Treatment selection after screening by Clearpath DS-120 lens fluorescence biomicroscope in diabetic patients

 Francesco Bandello,  Maurizio Battaglia Parodi,  Rosangela Lattanzio,  Federico Selvi
Departmento of Ophthalmology, University Vita-Salute, Scientific Institute San Raffaele, Italy

Purpose: To assess Clearpath DS-120 Lens Fluorescence Biomicroscope (Freedom Meditech, Germany) as screening test for the management of patients affected by diabetic retinopathy (DR).Methods: Overall, 101 patients were included in the study. All patients underwent an ophthalmologic examination complete of lens fluorescence biomicroscopy. A control group of 30 healthy subjects were considered for the data comparison.Results: Overall, the mean fluorescence ratio (FL-Ratio) was 0.2886 in the 101 patients and 0.1693 in the 30 controls (p 0.001). Mean FL-Ratio was 0.2488 , in the 34 diabetic patients without DR, and 0.3088 in the 67 diabetic patients with DR, with statistically significant difference (p  0.001). Receiver operating characteristic (ROC) analysis for presence of DR showed a cut-off of 0.2550 in patients under 50 years of age (sensitivity 92%, specificity 90%, AUC=0.961) and of 0.2650 in patients over 50 years of age (sensitivity 73.8%, specificity 73.5%, AUC=0.772). All the patients revealing FL-Ratio greater than 0.25 were more further investigated with fluorescein angiography and optical coherence tomography, showing that 22.8% had diabetic macular edema and 28.5% proliferative DR, and received specific treatment. In particular, 23 patients (18%) underwent intravitreal anti-VEGF, 8 patients (9%) received dexamethasone implant, and 47 patients (51%) required laser photocoagulation.Conclusion: FL-Ratio values can effectively separate patients affected by DR from control subjects. In addition, FL-Ratio greater than 0.25 suggests that patients are involved by DR, leading to a specific treatment algorithm. Our results indicate that Clearpath DS-120 Fluorescence Biomicroscope can be a useful screening test for DR.

 

 

544

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Retina

Long-term rescue of photoreceptors and vision following AAV-mediated corrective gene therapy delivered at intermediate disease stages in a canine model of RPGR X-linked retinitis pigmentosa

 William Beltran1,  Artur Cideciyan2,  Simone Iwabe1,  Malgorzata Swider2,  Mychajlo Kosyk2,  Gui-shuang Ying2,  James Shaffer2,  Wen-Tao Deng3,  Sanford Boye3,  Alfred Lewin4,  William Hauswirth3,  Samuel Jacobson2,  Gustavo Aguirre1
1Clinical Studies, University of Pennsylvania, USA
2Ophthalmology, University of Pennsylvania, USA
3Ophthalmology, University of Florida, USA
4Molecular Genetics & Microbiology, University of Florida, USA

Purpose: RPGR gene augmentation successfully rescues photoreceptors for up to 38 wks of age when delivered shortly after the onset of degeneration in a canine model of XLRP (Beltran et al. PNAS 2012). We now report on the long-term (~ 3 years) follow-up of dogs treated at early, and two intermediate stages of disease.

Methods: Seven XLPRA2 dogs that were treated with a unilateral subretinal injection (70-150 ul) of an AAV2/5 vector (1.51 x1011 vg/ml) carrying full-length human RPGR1-ORF15 cDNA under control of a hIRBP promoter at early (age=5 wks; ONL near normal thickness), early-intermediate (age=12 wks; ONL=~60% of normal) and late-intermediate (age=26 wks: ONL=~50-40% of normal) stages of disease were followed up to 164 wks of age. Retinas were examined in vivo by SD-OCT and ERG, and by histology/immunohistochemistry at termination. Visually-guided behavior of dogs from all three treatment stages was assessed in an obstacle avoidance course. A forced 2-choice Y maze was also used to test 2 dogs treated at late-intermediate disease.

Results: Statistically significant photoreceptor rescue was still detectable by OCT at ~ 130 weeks of age in the treated eyes of dogs from all 3 groups. Improved retinal (ERG) function and visually-guided behavior was seen in the treated eyes when tested between 130 and 164 wks of age.

Conclusions: This long-term and stable rescue of photoreceptor structure and function resulting in improved vision even when corrective gene therapy is delivered at more advanced stages of disease broadens the therapeutic window for intervention in patients with RPGR-XLRP.

 

 

776

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Drug Development

Prolonged release system for ocular administration of bevacizumab obtained by freeze-drying

 Susi Burgalassi1,  Susi Burgalassi,  Susi Burgalassi,  Daniela Monti1,  Silvia Tampucci1,  Cinzia Lavorini1,  Andrea Vento2,  Patrizia Chetoni1
1Department of Pharmacy, University of Pisa, Italy
2Ophthalmology Unit, Versilia Hospital, AUSL 12, Italy

Purpose

The use of intravitreal anti-vascular endothelial growth factor agents is increased dramatically over the last few years. This is due much to the successful use of off-label intravitreal bevacizumab for the treatment of exudative age-related macular degeneration, although intravitreal bevacizumab has been utilized to treat numerous ocular disorders. However, possible complications and side effects of the procedure of administration can occur. These may include eye pain, inflammation, infection, visual disturbances, but also more severe adverse events. The need to repeat the administration, even at an early date, greatly increases the risk of side effects. To reduce the number of applications, a prolonged release system of the drug could be very useful.

This work aimed at tuning solid formulations for ocular administration of bevacizumab to produce a sustained release of drug.

Methods

The formulations consisting of hydroxypropylmethyl cellulose matrices were obtained by freeze-drying of polymeric dispersions and were sterilized by gamma irradiation. The matrices underwent technological characterization for evaluating their hydration ability by solvent sorption time test and the release rate of drug by dynamic dialysis. The drug stability after production and sterilization processes was also evaluated. Finally, the tolerability was tested in albino rabbits.

Results

The matrices had sizes suitable for intravitreal or subconjunctival administration, hydrated slowly, and maintained drug stability. Furthermore, this vehicle was able to control the drug release: 2.0% bevacizumab was released over 4 days, and a release period of the loaded dose of some months could be expected. Moreover, they showed good tolerability in animals.

 

 

 

732

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Inflammation and Infection

In vitro activity of toyocamycin and novel thioinozine derivative as potential drugs against amphizoic amoebae - the causative agents of Acanthamoeba keratitis

 Lidia Chomicz1,  Marcin Padzik1,  Justyna Izdebska2,  Wanda Baltaza1,  Piotr Wroczynski3,  Jacek P. Szaflik2
1Department of Medical Biology, Medical University of Warsaw, Poland
2Department of Ophthalmology, SPKSO Ophthalmic Hospital, Medical University of Warsaw, Poland
3Pharmacy Department with the Laboratory Medicine Division, Medical University of Warsaw, Poland

Purpose. In the present study, we examined in vitro efficacy of toyocamycin and novel thioinozine derivative against clinical and environmental Acanthamoeba strains.

Methods. The corneal isolate of Acanthamoeba T4 deriving from patient with Acanthamoeba keratitis and Acanthamoeba castellanii Neff strain were cultured under bacteria-free condition at 26°C. In log growth phase, the amoebae were exposed to novel thioinozine derivative and toyocamycin. Different agent’s concentrations were applied and effects assessed after 24, 48, 72, 96, 120 and 144hrs. The status of surviving amoebae was examined, the overall amoeba number for 1ml of the culture calculated and percentage of cysts determined; all assays were repeated on twice; results were analyzed statistically.

Results and Conclusions. There was different amoebic sensibility to chemicals tested. The compounds influenced amoebic morpho-physiological status, overall amoeba numbers, the appearance of rounded, motionless forms of trophozoites. The strongest anti-Acanthamoeba efficacy against both strains was observed in assays with toyocamycin after 24-48hrs from this chemical application. Novel synthesized thioinozine derivative was not as effective as toyocamycin and inhibited development of amoebae after 96-120h exposition. The amphizoic amoebae are generally extremely resistant to different environmental factors and chemical agents. Results of our study showed that the corneal strain was less susceptible to the agents than Neff strain, however, the compounds indicated some amoebostatic influence upon both Acanthamoeba strains. It is noteworthy that no clear changes in cysts/ trophozoites ratio were found, thus the agent activity inhibitive for ability of these amoebae to transform into cysts cannot be excluded.

 

 

 

 

 

816

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Retina

Intra-vitreal Dexamethasone / Ozurdex implant in Retinal Vein Occlusions review of efficacy and safety in clinical practice 1, 2 & 3 year results

 Helena Cilliers,  Andleeb Zafar,  Lubna Razzaq
Machen Eye Unit, Warwick Hospital, South Warwickshire NHS Foundation Trust, UK

 

Purpose: To determine the efficacy and safety of 0.7mg intra-vitreal Dexamethasone/Ozurdex in patients diagnosed with retinal vein occlusions(RVO ́s). Whether there is any visual benefit in treating patients with longstanding 1yr RVO ́s, some with prior treatment of Ranibizumab/Avastin and/or Argon laser.

Method: 95 eyes had intra-vitreal Dexamethasone/Ozurdex treatment for RVO ́s since October 2011 with 1 year data by July 2015, 81 eyes 2 year data & 38 eyes 3 year data. This study includes patients with prior treatment of Ranibizumab/Avastin, Argon laser or combination treatment.

Retrospective review of patients treated at Warwick Hospital, UK. Demographics include types of RVO ́s and prior treatments. Findings after treatment: change in visual acuity(VA) & central macular thickness(CMT); morbidity from glaucoma, cataract and endophthalmitis.

Results: Age of patients 41 - 92 years, mean=71yrs at 1st treatment
. Male=46; Female=49. 
CRVO=42; BRVO=41; HRVO=12.


7 bilateral RVO ́s with 3 receiving treatment in each eye. 10 longstanding 1yr RVO ́s.

Previous treatment: Ranibizumab/Avastin 40=42%; Argon laser 20=21%; combination 18=19%.
19=20% with glaucoma/ocular hypertension (OHT); 73=77% were phakic.
 Completed data by July 2015 will be discussed in detail.

Conclusion: It seems intra-vitreal Dexamethasone/Ozurdex treatment modulates visual recovery, however contribution due to the natural recovery process must play a role. At some stage, inevitable chronic structural change results by which no further visual improvement is possible despite continued treatment. It would benefit to know exactly when this takes place, as it will determine decisions on future active intervention while taking into account treatment morbidity of intra-ocular procedures, glaucoma and cataract. Efficacious treatment within the best timeframe for visual improvement needs further randomised controlled study.

 

 

 

 

 

 

 

 

717

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Drug Development

Essential Polyunsaturated Fatty Acid-Phloroglucinol conjugates protect RPE and Neural Retina against All-trans-Retinal-induced Damages

 Aurelie Cubizolle1,  David CIA3,  Celine CRAUSTE2,  Laurent GUILLOU1,  Claire VIGOR2,  Thierry DURAND2,  Joseph VERCAUTEREN2,  Christian HAMEL1,  Philippe BRABET1
1INM team 01 vision, Inserm U1051, France
2IBMM, UM1 UMR5247, France
3Laboratoire de biophysique neurosensorielle, Inserm UMR1107, France

Purpose: We previously showed that phloroglucinol (PG) has a dual action against carbonyl and oxidant stresses from all-trans-retinal (atRAL) in retinal pigment epithelium (RPE). Disadvantage of PG is its low bio-availability. Therefore, we improve lipophilicity and reactivity with atRAL developing PUFA-O-alkylated-PG conjugates. Our objective is to assess their protective effect on RPE and neural retina (NR) against atRAL in-vitro and light-induced damages in-vivo (ABCA4KO mouse model).

Methods: ARPE19 cells are treated with PG derivatives for 1h and exposed to 25 µM atRAL for 4h. Similar co-treatments are applied to primary cultures of rat RPE and mouse NR. Cell viability (MTT assay) and flow cytometric (AnnexinV/PI) analysis are performed. Mitochondrial function is analyzed by oxygraphy and enzymatic activities. In parallel, acute stress induced by light (20000 lux, 2h) is applied to ABCA4 mice to evaluate PG-conjugates protection against photoreceptor degeneration.

Results: Monoisopropyl-phloroglucinol-DHA (PG-OIP-DHA) increases survival of ARPE19 (56.5%), RPE and NR primary cultures (37.5 to 90%). PG-OIP-DHA fully rescues ARPE19 cells from atRAL-induced late apoptosis and necrosis. Linoleic acid substitution has similar effect on cell viability, linolenic and ecosapentanoic acids show intermediate efficacy. PG-OIP-DHA and PG-OIP-LA rescue atRAL-induced damage on mitochondrial function. In-vivo we are now setting up best conditions of treatment.

Conclusion:  The protective effect of the PUFA-O-alkylated-PG conjugates is demonstrated in cultured RPE and neural retina challenged with a toxic dose of atRAL. Both PUFA and isopropyl are confirmed to be essential for the activity of these derivatives.  Currently we are testing in-vivo protection.

Financial disclosure: Yes

 

 

 

778

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Retina

OM-101 reduce the formation of fibrotic response associated to proliferative vitreoretinopathy

 Zeev Dvashi,  Yoel Greenwald,  Ran Stein,  Ayala Pollack
Ophthalmology, Kaplan Medical Center affiliated to the Hebrew University of Jerusalem, Israel

Purpose:

Proliferative vitreoretinopathy (PVR) is a scarring process that develops as a complication of retinal detachment (RD) and it is the most common cause of surgical failure upon RD treatment. PVR is a dynamic process characterized by the formation of fibrotic tissue on and under the detached retina, preventing the re-attachment of the retina. PVR is currently managed surgically with limited success. Since the hallmark of the pathological mechanism of fibrosis is not fully understood there are limited proven drugs to treat PVR . The goal of this study is to develop a new drug for PVR that will inhibit the fibrotic response.

Methods:

7-8 weeks old c57black mice underwent intravitreal injection of dispase (0 .3 units) to generate PVR in mice. 96 hours following dispase injection the mice were divided randomly to two groups, (10 mice for each group): treated with PBS or OM-101. After additional 96 hour mice were sacrificed, eyes enucleated and processed for histological and immunohistochemical analysis. (N=40).

Results:

Intravitreal injection of dispase causes RD and subsequently PVR in the control mice. In contrast mice that were treated with OM-101 developed RD with no fibrotic response and without the complication of PVR. The use of OM-101 maintains the normal morphology of the mice retina in compare to control mice and reduce fibrotic markers.

Conclusions

This study demonstrated that in this animal model OM-101 is an effective drug that halts the development of RD and prevents the occurrence of PVR

 

 

843

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Glaucoma

Clinical Evaluation of the Eye-to-visual-pathway Integrity of Glaucomatous Neurodegeneration Using 1.5T MR Imaging

 Kaya Nusret Engin1,  Nurten Turan Guner3,  Sibel Toreyen Bayramoglu3,  Ulviye Yigit2,  Onur Ozyurt4,  Muhittin Taskapili5,  Ahmet Agachan2,  Penbe Cagatay6
1Ophthalmology, Umraniye Education Research Hospital, Turkey
2Ophthalmology, Bakirkoy Education Research Hospital, Turkey
3Radiology, Bakirkoy Education Research Hospital, Turkey
4Biomedical Engineering, Bosphorus University, Turkey
5Ophthalmology, Beyoglu Goz Hospital, Turkey
6Biostatistics and Medical Informatics, Istanbul University, Turkey

Purpose: Accumulating data imply that glaucoma may actually represent a neurodegenerative disorder affecting the entire visual system. We evaluated retrobulbar glaucomatous damage with favorable techniques for 1.5T diffusion-tensor magnetic resonance (DT MR) imaging and we compared those techniques with clinical data in a large case series.

Methods: This Cross-sectional study included 130 eyes of 65 patients with primary open angle glaucoma. Patients with no known ocular or systemic concomitant disorders, neurological diseases, previous glaucoma surgeries, or antioxidant usage were selected. Optical coherences tomography and central visual field results of the subjects were recorded. Glaucoma analysis with optical coherences tomography (OCT) and standard automated perimetry (SAP) results of the subjects were recorded. DT MR analysis of optic nerves and radiations were performed, computing fractional anisotropy (FA), apparent diffusion coefficient (ADC), axial diffusivity (λ1) and radial diffusivity (λ┴). Correlation between the DT MR and clinical eye parameters of glaucomatous neurodegeneration were statistically evaluated.

Results: The correlations between diffusion parameters and age were highly significant. Optic nerve- fractional anisotropy and ipsilateral optic radiation-λ1 were significantly correlated with corneal thickness. Statistically significant correlations were found between ganglion cell complex and apparent diffusion coefficient, λ1, λ┴ of optic nerves.

Conclusions: Eye-brain connection in glaucoma can be evaluated with routine clinical instruments. Our study also revealed a limited correlation of retrobulbar glaucomatous neurodegeneration with ophthalmic damage. A better understanding of retrobulbar damage will enable us to develop more efficient strategies and a more accurate understanding of glaucoma.

 

 

810

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Basic Science

Targeting Receptor Interacting Protein-2 (RIP2) to Prevent Hyperglycemia-Mediated Inflammatory Signaling in Müller Cells

 Derrick Feenstra,  Susanne Mohr
Physiology, Michigan State University, USA

Purpose: Activation of the pro-inflammatory caspase-1/Interleukin-1β (IL-1β) pathway plays an important role in the development of diabetic retinopathy. In order to develop therapies targeting this pathway a better understanding of the mechanisms leading to caspase-1 activation under diabetic conditions is necessary. Receptor Interacting Protein-2 (RIP2) is a known activator of caspase-1 in other systems, and this study tests the hypothesis that caspase-1 is activated by (RIP2) under hyperglycemic conditions and by IL-1β signaling.

Methods: Primary human Müller cells (hMC), were isolated and cultured from retinas of non-diabetic donors. hMCs (1x106 cells) were treated with normal (5 mM) or high (25 mM) glucose for 48 hours or treated with recombinant IL-1β (2ng/ml) for 24 hours in the presence or absence of siRNA against RIP2. Caspase-1 activity and IL-1β release was measured.

Results: Caspase-1 activity was significantly increased by 53±3.0% (p0.05) in Müller cells treated with high glucose compared to those treated with normal glucose. siRNA against RIP2 decreased high glucose-induced caspase-1 activity from 16.8±0.3 to 11.9±0.7 pmol AFC/mg/min (p0.05). Accordingly, high glucose-induced IL-1β release was significantly attenuated from 6.2±0.3 to 1.4±0.9 pg/ml/mg (normal: 2.0±0.4 pg/ml/mg) in cells treated with siRNA against RIP2. IL-1β treatment led to increased caspase-1 activity (67.6±7.0%; p0.05). siRNA against RIP2 abrogated IL-1β-induced caspase-1 activity by 62±1.5% (p0.05).

Conclusion: Our results demonstrate that RIP2 indeed mediates hyperglycemia-induced caspase-1 activation in Müller and thus, might be responsible for the onset of retinal inflammation seen in diabetic retinopathy. Therefore, RIP2 represents a valid therapeutic target to treat diabetic retinopathy.

 

 

760

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Drug Development

Drug-Induced Ocular Side Effects of Clinical Importance to Ophthalmologists

 Rick Fraunfelder
Ophthalmology, University of Missouri, USA

Purpose:  To describe commonly reported side effects for topical or systemic drugs as they relate to the eye.

Methods:  Spontaneous reports collected at the National Registry of Drug-Induced Ocular Side Effects (www.eyedrugregistry.com)

Results:  Topiramate, hydroxychloroquine, herbal medicines, fluoroquinolones, vaccines and other agents can cause significant adverse ocular side effects.

Conclusion:  Clinicians should be aware of common adverse ocular events from frequently prescribed medications.

 

 

805

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Retina

TAK1 and PI3K inhibition decreases oxidative stress in RPE cells

 

 

 Mordechai Goldberg,  Michal Shapira,  Zeev Dvashi,  Zeev Dvashi,  Ayala Pollack,  Ayala Pollack
ophthalmology, Kaplan Medical Center affiliated with the Hebrew University Jerusalem., Israel

Purpose:

The accumulation of reactive oxidative species (ROS) due to oxidative stress, in retinal pigment epithelial (RPE) cells may change their morphology, damage blood retina barrier and can contribute to development and progression of age related macular degeneration (AMD). Several proteins were found to be involved in the accumulation of ROS including phosphatidylinositide 3-kinases (PI3K) and transforming growth factor β activated kinase (TAK-1). This study aims to investigate the role of PI3K and TAK1 in ROS accumulation and possible synergistic effect of these two proteins in RPE cells .

Methods:

Human ARPE-19 cells were treated with 5Z-7 oxozeaenol (TAK1 inhibitor) or LY294 (PI3K inhibitor) or combined treatment. Cells were exposed to H2O2 , washed, treated with 2`-7`-Dichlorodihydrofluorescein-diacetate (DCFH-DA) and subjected to Fluorescence Activated Cell Sorting (FACS).

Results:

In untreated RPE cells the level of positive DCFH-DA cells was less than 20% from the overall cell population. In cells treated with H2O2 it increased to 66%. In cells treated with TAK1 inhibitor or PI3K inhibitor it decreased to 45% and 60% respectively. Inhibition of TAK1 and PI3K decreased positive DCFH-DA cells to 35%.

Conclusions:

The main risk factor for development of dry AMD is oxidative damage. This study demonstrates that inhibition of TAK1 and PI3K results in an additive decrease of ROS accumulation in RPE cells upon oxidative stress. Thus, this outcome may be used as a therapeutic venue to reduce occurrence and progression of AMD upon oxidative damage.

 

 

 

808

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Inflammation and Infection

CORNEAL CROSS-LINKING AS AN ALTERNATIVE TREATMENT IN REFRACTORY ACANTHMOEBA KERATITIS

 Justyna Izdebska1,  Lidia Chomicz2,  Marcin Padzik2,  Wanda Baltaza2,  Jacek P. Szaflik1
1Department of Ophthalmolgy, Medical University of Warsaw, Poland
2Department of Medical Biology, Medical University of Warsaw, Poland

PURPOUSE: The aim of the study was to determine whether the corneal cross-linking (CXL) procedure can be an effective method of treatment of acanthamoeba keratitis (AK) unresponsive to medical therapy.


 METHODS: We analyzed 10 cases of diagnosed AK that were treated in our department between 2012 -2014. Our group consists of 10 patients (4 women, 6 men) aged from 26 to 41y.o who went to our hospital after 3-37 days of the first symptoms often after an earlier unsuccessful conservative treatment. Parasitological study included: preparation of corneal scrapings preparations and cultured in vitro. All eyes were treated with a topically applied: propamidine isethionate (0,1%), a combination of chlorohexidine (0.02%) and polyhexamethylen biguanide (PHMB, 0.02%), neomycin for at least 14 days without significant clinical improvement. In all of these cases the CXL was performed. After CXL topical treatment was continued for several weeks.

RESULTS. Cysts and/or trophozoites of Acanthamoeba were detected in all 5 patients, once the corneal scrapings and/or in cultures after 2-7 days after transplantation. In all cases, the corneal inflammation due to acanthamoeba infection was significantly diminished or retreated. In 2 eyes the urgent keratoplasty was performed due to significant corneal thinning without melting. 3 patients will need corneal graft to improve vision due to central corneal scar. In 5 cases patients will need no further surgical intervention.


 CONCLUSIONS. Corneal cross-linking can be effective treatment in refractory AK. In some cases patients can avoid corneal graft, in others keratoplasty can be performed in as a non-emergency procedure.

 

 

815

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Basic Science

KinostatTM prevents cataracts in diabetic dogs

 

 Peter Kador1,2,  Milton Wyman1,  Manley Paulos1,  the Kinostat Trial Study Group3
1Research, Therapeutic Vision, Inc., USA
2College of Pharmacy, University of Nebraska Medical Center, USA
311 Eye Centers, Veterinary Ophthalmologists, USA

Purpose: A majority of dogs develop blinding bilateral cataracts within 6 months after diagnosis of diabetes mellitus (DM). Here, we present an interim analysis of a randomized masked placebo controlled clinical trial (1/3 placebo) of the topical aldose reductase inhibitor KinostatTM that is being conducted at 11 centers across the United States.

Methods: 135 dogs are being evaluated by board certified veterinary ophthalmologists at the time of enrollment and then at 1, 2, 3, 6 and 9 months. The dog’s owners administer the topical formulations TID. Dogs not developing cortical cataracts during the 9-month period are then given KinostatTM with ophthalmic evaluations required at 6-month intervals.

 

Results: Newly diabetic dogs of all sizes, breeds, and sex with only equatorial vacuoles of less than 360o present and no other ocular disease were recruited. The results, to date, confirm the initial proof of concept study (Vet. Ophthlamol. 13:363-8, 2010) that daily administration of KinostatTM to diabetic dogs significantly prevents cataract formation for up to 6-years. A required toxicology study found that daily application of KinostatTM at doses of up to 5x the recommended doses did not induce any direct local or systemic toxic effects in any of the tissues examined.

 

Conclusion: KinostatTM is the first drug to prevent the clinical development of diabetic cataracts and reduce the need for cataract surgery. Because KinostatTM meets an unmet medical need, the FDA has granted KinostatTM a fast-track Minimum Use in a Major Animal Species designation.

 

 

 

917

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Basic Science

A novel target for therapy development in optic neuropathies – a second life for presenilins?

 Peter Koulen
Vision Research Center, Department of Ophthalmology, and Department of Basic Medical Science, University of Missouri - Kansas City, School of Medicine, USA

Loss of visual function in optic neuropathies is characterized by the degeneration of retinal ganglion cells (RGCs). Cell death of RGCs is preceded by cellular calcium dyshomeostasis and toxicity caused by chronically elevated intracellular calcium concentrations. Control of calcium signaling pathways has become the target of related therapy development efforts. Others and we recently described a new role for presenilin proteins. In addition to their function in amyloid precursor protein processing, presenilins control the intracellular calcium concentration by regulating the activity of intracellular calcium release channels. The present study tested the hypothesis that modulation of the presenilin protein concentration leads to RGC protection. 

The viability of murine RGCs was measured in response to chronic L-glutamate-mediated toxicity using immunocytochemistry assays. After altering the concentration of presenilins in RGCs changes in intracellular calcium ion signaling were measured using electrophysiology, calcium imaging and pharmacological control of intracellular calcium channel activity.

Both presenilin 1 and 2 are expressed by RGCs. Knockdown of presenilin 1 significantly increased viability of both isolated cultured RGCs and of organotypic cultures. This was paralleled by significantly attenuated calcium release from intracellular stores. Overexpression of presenilin 1 elicited potentiated calcium release from intracellular stores and decreased viability of RGCs. Modulation of presenilin 2 generated similar responses.

Presenilins control calcium release from intracellular stores and thereby affect cellular viability as a function of cellular calcium dyshomeostasis following injury or resulting from disease processes. These mechanisms of action therefore represent novel potential targets for therapeutic intervention and drug development in optic neuropathies.

 

 

577

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External Eye Diseases

The In Vitro and In Vivo Antibacterial Evaluation of Brilacidin

 Regis P. Kowalski1,  Eric Romanowski1,  Kathleen Yates1,  Robert Shanks1,  Francis Mah2
1Ophthalmology, University of Pittsburgh, USA
2Ophthalmology, Scripps Health, USA

Purpose: Brilacidin (BRI) (PMX30063) is the first anti-infective in a new class of defensin mimetics. The goals of the study were to evaluate the efficacy of BRI as an ocular anti-infective.

Methods: In vitro: MICs were determined for ocular isolates (n = 25/ bacteria) of Ciprofloxacin-Susceptible-Staphylococcus aureus-(CSSA), Ciprofloxacin-Resistant-Staphylococcus aureus- (CRSA), CS-Staphylococcus- epidermidis- (CSSE), CR-Staphylococcus- epidermidis- (CRSE), Streptococcus- pneumonia-(SP), Streptococcus-viridans- (SV), Moraxella- (MS), Haemophilus-influenzae- (HI), Pseudomonas-aeruginosa- (PA), and Serratia- marcescens-(SM). In vivo: In 24 NZW rabbits, the left cornea was abraded and the right remained intact. The corneas were infected with 1000 CFU of MRSA. Four hrs passed. Topical drops were administered (every 15’ for 5 hrs) to 4 respective groups: A) BRI 0.5%, B) Vancomycin (VAN) 5%, C) saline (SAL), and D) no treatment (baseline CFU). One hr after treatment the corneas were harvested for CFU. Results: In vitro: Data (μg/ml) is expressed as MIC50, MIC90, and MIC Range. CSSA(0.25, 0.25, 0.125-0.5); CRSA(0.25, 0.5, 0.125-1.0); CSSE(0.125, 0.25, 0.03125-0.25); CRSE(0.125, 0.25, 0.03125-0.25); SP(1, 1, 0.5-128); MS(4, 64, 0.5-128); HI(8, 8, 2-32); PA(4, 4, 0.5-8); SM(8, 32, 0.25-32). In vivo: For abraded corneas, VAN=BRISal for reductions in MRSA CFU. BRI demonstrated a 99.9% reduction compared to baseline CFU. For intact corneas, VANBRI SAL. BRI=baseline counts suggesting the corneal epithelium acts as a barrier for penetration. Conclusions: BRI demonstrated broad spectrum in vitro activity against ocular pathogens. BRI was equally efficacious as VAN in a MRSA keratitis model only when the corneal epithelium was removed.

 

 

 

739

--------------------------------------------------------------------------------------------

Retina

Ocular manifestations and choroidal thickness measured by Swept-Source OCT (SS-OCT) in patients with familial hypercholesterolemia (FH) treated with -------------------------------------------------------------------------------------------- intensive statin therapy.

 Félix Alexander Manco Lavado1,3,  Maria Rosalba Ramoa Osorio1,3,  Gonzalo Diaz Soto2,  Maria Isabel Lopez Galvez1,3,  Lucia Manzanas Leal1,3
1Ophthalmology, Hospital Clinico Universitario de Valladolid, Spain
2Endocrinology, Hospital Clinico Universitario de Valladolid, Spain
3Ophthalmology, IOBA, Spain

Purpose: High levels of cholesterol have been related to macular degeneration. In animal models hypercholesterolemia damages the neurosensory retina and induces an increase in thickness of the choroid and sclera. The purpose of this study is to evaluate the anatomical findings in FH patients treated with -------------------------------------------------------------------------------------------- intensive statin therapy using SS-OCT.

Methods: We designed a descriptive, cross-sectional and comparative study of 18 FH patients treated with statins versus 18 healthy controls. All patients underwent a complete ophthalmic examination. Primary outcomes were the presence of typical ocular manifestations of FH, and the quantitative and qualitative changes in retina and choroid on SS-OCT in FH patients compared to controls.

Results: Mean age of FH patients was 54.0±11.9 years-old, 50% were male. Mean LDL-c level at diagnosis was 260.6±47.6mg/dl, but 133.7±34.0mg/dl twenty-years after treatment. Mean best-corrected visual acuity was 0.01±0.04 logMAR, corneal arcus was found in 61.1%, xanthelasmas in 11.1% and one patient showed vascular narrowing in the fundus. Neither lipemia retinalis nor cholesterol crystals were found. There were no qualitative changes in the retina analysed by SS-OCT. Mean subfoveal choroidal thickness (CT) was 265.17±96.71μm, nasal-CT 210.90±88.70μm and temp---------------------------------------------------------------------------------------------CT 208.20±90.43μm. In control group, the mean age was 42.3±11.9 years and the mean LDL-c level was 110.6±45.2mg/dl. Choroidal and retinal thickness was within the normal range. No differences were found between both groups.

Conclusions: The intensive statin therapy prevents and reduces the ocular damage of hypercholesterolemia. The choroidal and retinal thickness in treated FH patients measured by SS-OCT is within the normal range.

 

 

562

--------------------------------------------------------------------------------------------

Drug Development

Special requirements for unpreserved multi-dose ophthalmic formulations

 Degenhard Marx,  Matthias Birkhoff,  Rolf-Peter Hummel
Resarch & Development, Aptar Radolfzell GmbH, Germany

The increasing significance of unpreserved formulations in eye care products triggers questions regarding what scientists in the pharmaceutical industry need to consider for an efficient and successful development. A first consideration is that in Europe artificial tear products are subject to the European Medical Device Directive, while combination products containing an active pharmaceutical ingredient are under supervision of the respective national drug regulating authorities. Conventional preserved eye drops rely on preservatives to ensure microbial integrity during storage, transportation and the in-use period. The related requirements for such preserved products are well established. On the other hand, no guidance documents are in place for unpreserved multi-dose systems. Preservative-free systems make use of some mechanical barriers and/or oligodynamic elements (e.g. silver ions) in order to maintain microbial integrity during storage, transportation and in use period. For a successful development, the formulation must be stable and remain sterile for the entire life span of the product. Drop size is hard to control but should be similar to marketed products. Authorities will certainly challenge the microbial barrier functions of the container closure system which needs to be demonstrated using suitable microbial challenge tests. Transport simulation and rough handling will have minor impact on conventional multi-dose bottles, but it may impair proper function of preservative-free systems, this again requires thorough testing. Easy and convenient handling has to be demonstrated (e.g. acceptable actuation force).

Conclusion: For the successful development of unpreserved multi-dose eye drops appropiate parameters need to be considered and close co-operation with the supplier of the CCS is mandatory.

 

 

813

--------------------------------------------------------------------------------------------

Basic Science

Normalising eNOS function to facilitate repair of the ischaemic retina

 Denise McDonald
Centre for Experimental Medicine, Queen's University Belfast, UK

In ischaemic retinopathies, an abnormal response to growth factor stimulation leads to the misdirection of reparative angiogenesis away from the hypoxic retina leading to sight-threatening intravitreal neovascularisation (NV). Therapeutic strategies which could reverse this trend would be extremely beneficial. Endothelial nitric oxide synthase (eNOS)-derived NO plays an important role in promoting vascular growth and survival, a function which is dependent upon the cofactor, tetrahydrobiopterin (BH4).   Previously, we have shown that oxidative stress depletes retinal BH4 levels, reduces NO production and exacerbates vascular closure, the prelude to ischaemic injury. Thus, here we investigated if supplementing BH4 levels can protect the retina from oxidative insult and normalise vascular growth.

Endothelial specific eNOSGFP transgenic mice at postnatal day 7 were subjected to 2-5 days, and retinal microvascular endothelial cells (RMEC) to 24h hyperoxia to induce oxidative damage. Supplementation was achieved with sepiapterin and elevated BH4 levels confirmed by HPLC, NO production was measured by nitrite and NOS activity and nitrotyrosine (NT) by western blotting. Proliferation was determined by BrdU labelling and lectin staining of retinal flat mounts was used to determine the extent of vessel closure before and after treatment.

Hyperoxia exposure in vitro and in vivo deleted BH4 levels, diminished NO production, elevated free radical production and negatively impacted EC proliferative ability.   These effects were corrected by BH4 supplementation which improved NO production, enhanced EC proliferation and increased vessel coverage in eNOSGFP animals. Thus, BH4 supplementation reverses oxidative stress-induced endothelial damage and preserves vascular integrity in the neonatal retina.

 

 

 

809

--------------------------------------------------------------------------------------------

External Eye Diseases

Eye pain and secondary headache in in the course of infection caused by Demodex

 Izabela Chudzicka-Strugala1,  Walenty Chudzicki2
1Department of Microbiology, Poznan University of Medical Sciences, Poland
2Ophthalmology, Private Eye Clinic, Poland

Demodex is an external parasite. Exists in hair follicles and sebaceous glands of the skin. Changes are most often located in the nose region, around the eyes, on the forehead, chin. May also ride on other parts of the body for example hands and foot skin. Can also induce a chalazions, or acne rosacea. The incidence of infection increases with age. Demodex can induce blepharitis or blepharoconjunctivitis.

Objective: Demonstration of the incidence of eye and head pain in the course of infection with Demodex.

Material and methods: The investigated group included 320 patients (180 women and 140 men 25-82 years old) with blepharitis or blepharoconjunctivits with eye pain, swelling of the eyelids, and haedache. Each patient was previously treated with long-term neurological ophthalmologist and no effects. The diagnostics material included parasitology nad microbiology examination. Were epilated 4 eyelashes from each eyelid and also swab from conjunctivita for bacteriological culure. Eyelashes were placed on Glass sidle in 10% KOH. Under the microscope (magnification 100X) was observe presence or no Demodex (eggs, nimphs , mature form, larvae form). To culturing used blood agar and next sellective agar.

Results: In 290 (90%)( 160 (88,9%) women and 130 (93%) men) patients showed the presence of occurrence Demodex. In 30 (9%) patients, 14 (47%) women and 16 (53%) men were S. aureus bacterial co-infection or S.epidermidis.

Conclusion: 1. Patients with chronic blepharitis or blepharoconjunctivitis with pain eyes should be taken after the ophtalmic examination make parasitological and bacteriological: 2. Eye strain in the course of Demodex cause secondary headaches temp-------------------------------------------------------------------------------------------- and frontal; 3. Exact microbiological diagnostics will reduce the costs of treatment.

 

 

924

--------------------------------------------------------------------------------------------

Retina

Evaluation of the safety of repeated subthreshold micropulse yellow laser photocoagulation in diabetic macular edema treatment

 Elena Pedanova,  Dmitry Buryakov,  Galina Kachalina,  Leonid Kryl
Department of laser surgery, Federal budget state institution "The academic Fyodorov Eye microsurgery complex of the Ministry of public health of Russia", Russia

Background. Subthreshold micropulse yellow laser photocoagulation (SMYLP) have demonstrated its efficacy in diabetic macular edema (DME) management. High selectivity and low power parameters of SMYLP result in significantly reduced retinal damage. One of the limitations of SMYLP is transient therapeutic effect, which requires repeated laser treatment. There are no available studies aimed to assess the safety of repetitive SMYLP.

Purpose. To evaluate the safety of subthreshold micropulse laser 577-nm photocoagulation after repeated laser treatment sessions.

Material and methods. 17 patients (31 eyes) with DME who underwent three sessions of SMYLP with 1 month interval were examined at the baseline, before each treatment session and at 3-month follow-up. Retinal structure changes and functional outcomes were assessed.

Results. At 3-month follow-up mean central retinal thickness decreased from 405.34±107.22 to 378.65±111.15 (p0.05), best corrected visual acuity and central retinal sensitivity improved from 0.56±0.20 to 0.62±0.30 and from 14.53±4.20 dB to 16.13±2.50 dB respectively (p0.05). Autofluorescence in short-wave and infrared wavelengths didn`t reveal laser induced hyperfluorescence, which was considered as lack of pigment epithelium and choriocapillaris layers damage after treatment. Optical coherence tomography didn`t show any laser induced changes of neurosensory retina at any follow-up visit in each case. Microperimetry data confirmed absence of central field scotomas at final follow-up.

Conclusions. Three repeated sessions of SMYLP treatment of DME, appears to be a safe therapy technique that is not accompanied with retinal and choriocapillaris damage and functional disturbances.

 

 

807

--------------------------------------------------------------------------------------------

Drug Development

A Novel Ex Vivo Model to Evaluate the Role of Müller Cells in Retinal Drug Delivery

 Karen Peynshaert,  Thomas Martens,  Stefaan De Smedt,  Kevin Braeckmans,  Katrien Remaut
Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Belgium

Purpose: To develop a bovine ex vivo retinal model that, in contrast to existing models, keeps the vitreoretinal barrier intact. This model would allow us to evaluate the influence of particle physicochemistry on their ability to cross the vitreoretinal interface after intravitreal injection.

Methods: Bovine eyes are obtained from the abattoir. The retina – with vitreous attached - is detached from the RPE and placed on a Transwell® filter, after which supplemented Neurobasal medium is added below the filter. Mitotracker Deep Red (stains Müller cells) and FM 1-43 lipid dye is added to the medium and intravitreally injected prior to incubation overnight. Images are taken at multiple z-levels using a Nikon 60x NIR Apo water dipping objective with a Nikon C1si confocal microscope.

Results: The Mitotracker staining of the explant confirms that the ganglion cell layer is virtually completely occupied by Müller cell endfeet. Moreover, the vitreoretinal interface of bovine eyes greatly resembles human physiology. The lipid dye FM 1-43 outlines every Müller cell in this layer and clearly stains veins running through the retina. Following intravitreal injection of polystyrene particles, some colocalize with Müller cells, suggesting these cells could form a transport route across the retina. However, the distance of injection from the retina and nanoparticle concentration should be optimized.

Conclusions: We developed a model that maximally mimics human ocular physiology. We suggest that this model can serve as a set-up to aid in the evaluation and design of ocular non-viral gene delivery vectors with the retina as their target.

 

 

904

--------------------------------------------------------------------------------------------

Retina

Can AMD be prevented? The possible role of hydroxichloroquine in preventing AMD.

 Joseph Pikkel
Ophthalmology, Ziv Medical Center, Israel

Purpose: To investigate a possible structural difference in the retina of Plaquenil (hydroxychloroquine) treated patients as an explanation for the protective effect of this medication against age- related macular degeneration (AMD).

Design: Retrospective comparative study

 Methods: From one clinical ophthalmology practice, 54 eyes of 27 Plaquinil treated rheumatoid arthritis patients (study group), 40 eyes of 20 healthy similar aged individuals (control group I) and 22 eyes of 11 non-Plaquinil treated rheumatoid arthritis patients (control group II) were recruited from a clinical practice. All participants had undergone full clinical ophthalmic evaluation, central visual field test, and macular ocular coherence tomography (OCT). Thickness of the retinal outer band (consisting of the Bruch`s membrane and retinal pigment epithelium layer) were measured by OCT.

 Results: The mean thicknesses of the outer band of the retinal pigment epithelium layer were 60.4+7.4, 43.3+2.7, and 39.7+3.6 microns for the study group, control group I, and control group II, respectively. P values for differences in mean thicknesses were  0.0001 between the study group and each of the control groups, and 0.086 between the two control groups.

 `Conclusion: Treatment with Plaquenil was associated with increased thickness of the outer band of the retinal layer. This finding may explain the protective effect of Plaquenil against age- related macular degeneration (AMD).

 

 

916

--------------------------------------------------------------------------------------------

External Eye Diseases

Topical steroids protects the lacrimal functional unit of dry eye patients from desiccating stress

 

 

 

 Jose Pinto-Fraga1,2,  Alberto Lopez-Miguel1,2,3,  Maria Jesus Gonzalez-Garcia1,2,  Itziar Fernandez1,2,  Alberto Lopez-de-la-Rosa1,  Amalia Enriquez-de-Salamanca1,2,  Michael Stern4,  Margarita Calonge1,2
1IOBA (Institute of Applied OphthalmoBiology), University of Valladolid, Spain
2CIBER-BBN, Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, Spain
3VISION I+D ,SL, IOBA, Spain
4Allergan Inc, Allergan, USA

Purpose: To assess the efficacy of topical 0.1% fluorometholone for protect the lacrimal functional unit in moderate-to-severe dry eye disease (DED) patients when exposed to adverse environments.

Methods: Single-center, double-masked, parallel clinical trial (clinicaltrials.gov Identifier # NCT02051023). Participants randomly received topical 0.1% Fluorometholone or polyvinyl alcohol for 22 days. Corneal and conjunctival staining, conjunctival hyperemia, tear break-up time (TBUT), tear osmolarity and Symptom Assessment in Dry Eye questionnaire (SANDE) were assessed during baseline (V1), at day 21 before and after exposure to controlled adverse environment (V2 and V3), and at day 22 after 24 hours from the adverse exposure (V4). Main outcomes measures were the percentage of patients showing an increase ≥1 point in corneal staining and the percentage with a reduction ≥2 points in SANDE score, at V3 and V4.

Results: After 21-day treatment, improvements for corneal and conjunctival staining, conjunctival hyperemia, and TBUT were significantly higher (p≤0.03) for the FML group than for the LIQUIFILM group. No changes in osmolarity were observed. After the adverse exposure the LIQUIFILM group showed a significant (p=0.03) higher percentage of patients with a ≥1 grade increase in corneal staining than the FML group (63.1%, 95% confidence interval (CI): 38.6-82.7; 23.8%, 95% CI: 9.1-47.5, respectively). No significant differences between groups were found at V4. No adverse events were observed throughout the whole study.

Conclusions: Three-week topical 0.1% fluorometholone therapy is safe and effective to reduce ocular surface signs in DED patients and to prevent DED exacerbation in patients exposed to adverse conditions.

 

 

 

 

 

 

762

--------------------------------------------------------------------------------------------

Retina

Choroidal neovascularization in highly myopic elderly patients: A special form of choroidal neovascularization?

 Maria Rosalba Ramoa Osorio1,2,  Felix Alexander Manco Lavado1,2,  Ana Belen Haro Alvarez1,  Maria Isabel Lopez Galvez1,2,  Ignacio Alonso de la Fuente1,2
1Ophthalmology, Hospital Clinico Universitario de Valladolid, Spain
2Ophthalmology, IOBA, Spain

Purpose: To evaluate the safety and efficacy of intravitreal bevacizumab (IVB) in the treatment of high myopic choroidal neovascularization (CNV) in patients over 60 years of age.

Methods: We retrospectively reviewed ophthalmic medical records of all myopic eyes with CNV treated with IVB as first-line with a pro-re-nata (PRN) regimen and 12 months of follow-up. Only patients over 60 years of age and with a complete baseline ophthalmologic examination, including best-corrected visual acuity (BCVA), fundus examination, fluorescein angiography and optical coherence tomography were included in the study. An initial injection of IVB (1.25mg/0.05mL) was administered, and extra doses of IVB were used when persistent or additional retinal exudation was observed. Patient`s general clinical records were reviewed for systemic adverse events related to drug therapy.

Results: 21 eyes of 21 patients were included, 15 women (71.43%) and 6 men (28.57%) with a mean age of 66.27±5.47 and 68.17±4.14 years, respectively. Only 3 patients (14.29%) gained more than or equal to 10 letters of visual acuity at twelve months. 13 patients (61.9%) maintain visual acuity, and 5 patients (23,81%) lost at least 5 letters. The mean number of injections was 2.48±1.4 during the follow-up period. Regarding safety, two patients suffered angor pectoris during treatment (at 7 and 30 days after the injection). Both patients were at high risk for cardiovascular events.

Conclusion: IVB is useful in myopic CNV, but functional outcomes in patients older than 60 years are poor and visual gains are rare. Further studies are needed in this population.

 

 

584

--------------------------------------------------------------------------------------------

Inflammation and Infection

The In Vitro Evaluation of the Virucidal Efficacy of Povidone-Iodine against Multiple Ocular Adenoviral Types

 Eric Romanowski,  Kathleen Yates,  Robert Shanks,  Regis Kowalski
The Charles T. Campbell Ophthalmic Microbiology Laboratory, UPMC Eye Center, Department of Ophthalmology University of Pittsburgh, USA

Purpose: Povidone-iodine (P-I) has been proposed as a topical antiviral for adenoviral ocular infections. Several concentrations have been tested for potential clinical use: 5% (Betadine for EKC Study, www.betadineforekc.com); 2% (NCT01179412); 0.4% (as part of the 0.4% P-I/0.1% dexamethasone combination, NCT01470664). The goal of this study was to determine the in vitro efficacy of the clinically proposed concentrations of P-I against a panel of common ocular Ad types. Methods: The virucidal efficacy of 5%, 2%, 0.4%, and 0.001% P-I was determined for extracellular virus of types Ad3, Ad4, Ad5, Ad7a, Ad8, Ad19, and Ad37. Ad/P-I mixtures were incubated at 33oC. At 1, 5, 15, and 60 minutes of incubation, aliquots were removed. Standard viral titrations were carried out to determine the amount of virus present. Results: Virucidal (99.9%) decreases in Ad titers were demonstrated for 5%, 2%, and 0.4% P-I at 1m for Ad3, Ad4, Ad5, Ad7a, and Ad8. For Ad19, virucidal decreases were demonstrated at 60m for 5%, 1m for 2%, and 15m for 0.4% P-I whereas for Ad37, virucidal decreases were demonstrated at 60m for 5% and 2%, but 1m for 0.4% P-I. No virucidal decreases demonstrated for 0.001% P-I at any time point for any serotype. Conclusions: Tested P-I concentrations were virucidal at 1 minute for most of the ocular serotypes tested. However, the time to achieve a virucidal decrease for the EKC types Ad19 and Ad37 increased for several P-I concentrations. The efficacy of P-I in the treatment of ocular adenoviral infections may be type dependent.

 

 

775

--------------------------------------------------------------------------------------------

Retina

Objective perimetry based on chromatic multifocal pupillometer for treatment follow-up and diagnosis in patients with retinal and macular dystrophies

 Ygal Rotenstreich1,2,  Ygal Rotenstreich,  Ron Chibel,  Ron Chibel1,2,  Soad Hajyahia1,  Soad Hajyahia,  Daniel Ben-Ner1,  Mohamad Omar Mhajna1,  Michael Belkin1,2,  Ifat Sher1,2
1Goldschleger Eye Institute, Sheba Medical Center, Israel
2Sackler Faculty of Medicine, Tel Aviv University, Israel

Purpose: Objective non-invasive perimetry and diagnosis in healthy subjects and patients with macular and retinal degeneration using an infrared chromatic multifocal pupillometry.

METHODS: A multifocal chromatic pupillometer was used to record pupillary responses (PR) to red and blue light stimuli (peak 485 nm and 640 nm, respectively) presented by 76 LEDs at different points of the 18 degree visual field (VF). PR amplitude, constriction velocity (CV) and the time of maximal constriction velocity (TMCV) were measured in 17 retinitis pigmentosa (RP) patients, 5 Vitelliform Macular Dystrophy patients and 26 age-matched controls. Pupillometer results were compared with subjective Humphrey 24-2 and Goldmann perimetry.

RESULTS: RP patients demonstrated significantly longer TMCV and reduced amplitude and CV in majority of locations. The variability in TMCV recorded at different VF locations was significantly higher in RP patients compared with controls in response to the red stimulus (p0.0001). Tight correlation was observed between severity of VF loss determined by the chromatic Goldmann and the variability of TMCV (R=0.77). Macular dystrophy patients demonstrated significantly lower amplitude and CV in response to the red stimulus and nearly normal PR to the blue stimulus in majority of VF locations. High consistency was observed in PR recorded in serial testing (P0.001, R=0.74 for red and P0.001, R=0.683 for blue, n=870).

CONCLUSIONS: This study demonstrates the feasibility of using the multifocal chromatic pupillometer for objective non-invasive differential diagnosis, assessment of VF defects and phenotype characterization.

 

 

 

804

--------------------------------------------------------------------------------------------

Retina

A minimally invasive adjustable blunt injector for posterior segment delivery of drugs and cell therapy

 Ygal Rotenstreich1,2,  Adi Tzameret1,2,  Sapir Kalish1,  Itay Levy5,  Avraham Treves3,  Arnon Nagler2,4,  Michael Belkin1,2,  Mordechai Rosner1,2,  Iris Moroz1,2,  Shlomo Margel5,  Ifat Sher1,2
1Goldschleger Eye Institute, Sheba Medical Center, Israel
2Sackler Faculty of Medicine, Tel Aviv University, Israel
3Center for Stem Cells and Regenerative Medicine, Cancer Research Center, Sheba Medical Center, Israel
4Hematology Division, Sheba Medical Center, Israel
5Department of Chemistry, Bar-Ilan Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Israel

Purpose: Treatment for posterior segment diseases is limited by the challenge of safe delivery of therapeutics in effective dosages in proximity to the retina and choroid. Here we investigated the feasibility of using a novel minimally-invasive adjustable blunt injector for delivery of pharmaceuticals and stem cells across the extravascular spaces of the choroid.

Methods: A novel system comprised of a syringe with a blunt needle and an adjustable separator was developed. New Zealand White rabbits (n=20) were injected with stem cells, indocyanine green (ICG) or iron oxide nanoparticles (NPs). No immunosuppressants were used. Cadaver pig eyes were injected with ICG. The efficacy and safety of the surgical procedure were determined using Optical Coherence Tomography (OCT), Electroretinogram (ERG) and histopathology.

Results: Injected cells and pharmaceuticals were identified across the extravascular spaces of the choroid, covering over 70 percent of the sub retinal surface, and could be traced in the posterior eye for at least two weeks following injection. No retinal detachment, choroidal hemorrhages, inflammation or changes in retinal function were detected.

Conclusions: Targeting pharmaceuticals and cells to the posterior segment can be achieved using a novel injection platform in a safe and reproducible manner. Therapeutics are placed in close proximity to the retina as a thin layer, without insertion of surgical instruments under the macula, with no retinal detachment or choroidal hemorrhage. This new transplantation system is predicated to increase the therapeutic effect and safety of pharmaceuticals and advanced therapies for posterior segment diseases.

 

 

728

--------------------------------------------------------------------------------------------

Drug Development

Nanoparticle cross-linked collagen shields for ocular drug delivery

 Yosra Agban1,2,  Lily Lian3,  Ali Seyfoddin2,  Sujay Prabkar3,  Ilva D. Rupenthal1
1Buchanan Ocular Therapeutics Unit, Department of Ophthalmology, New Zealand National Eye Centre, The University of Auckland, New Zealand
2School of Pharmacy, The University of Auckland, New Zealand
3Leather and Shoe Research Association of New Zealand, Palmerston North, New Zealand

Glaucoma is a common progressive eye disease and remains the second leading cause of blindness. Treatment involves frequent administration of IOP lowering eye drops often resulting in poor patient compliance. To overcome this problem, this study aimed to develop a novel nanoparticle cross-linked collagen shield for sustained pilocarpine delivery.

Three metal oxide nanoparticles (NPs), titanium dioxide (TiO2), zinc oxide (ZnO) and polyvinylpyrrolidone capped zinc oxide (ZnO/PVP), having antimicrobial properties and potential anti-angiogenic effects, were evaluated for their cytotoxicity on primary human corneal epithelial cells (HCEC) before being used as cross-linking agents. Collagen shields were prepared by the solvent casting method and characterised in terms of transparency, mechanical strength and mucoadhesion. The shield with the best properties was then loaded with pilocarpine and in vitro release studies were performed. Finally, the effect of additional UV irradiation on mechanical properties and drug release was investigated.

ZnO/PVP NPs were the least toxic and at a ratio of 1:1 w/w (collagen: ZnO/PVP NPs) produced films with increased mechanical and mucoadhesive properties compared to plain collagen shields. The concentration of zinc ions released from cross-linked shields over time was below the IC50 (23.2 µg/ml), rendering them safe for ocular use. Pilocarpine release was controlled over a period of 14 days, with the application of UV light further decreasing the release rate of both zinc ions and drug.

In conclusion, collagen shields cross-linked with ZnO/PVP successfully delivered pilocarpine over prolonged periods of time offering a novel sustained release option for glaucoma treatment.

 

 

777

--------------------------------------------------------------------------------------------

Retina

in vitro and in vivo efficacy on retinal neo-vascularization of an innovative broad-range anti-angiogenic synthetic peptide (uparant)

 Dario Rusciano1,  Paola Bagnoli2,  Marco Presta3,  Francesco Semeraro4,  Massimo Dal Monte2,  Maurizio Cammalleri2,  Sara Rezzola3,  Mirella Belleri3,  Vincenzo Pavone5,  Helder Andre6
1Scientific, Sooft Italia SpA, Italy
2Biology, University of Pisa, Italy
3Molecular and Tranlsational Medicine, University of Brescia, Italy
4OIphthalmology, University of Brescia, Italy
5Chemical Science, Federico II University, Italy
6Clinical Neurosciences, Karolinska Institute, Sweden

Purpose: To show preclinical efficacy on retinal neoangiogenesis of a new concept molecule (Uparant), designed to inhibit the urokinase-type plasminogen activator receptor and able to prevent the angiogenic response elicited by a broader range of angiogenic factors besides VEGF.

Methods: Different concentrations of Uparant were given: in vitro to vascular endothelial cells and tissue cultures subjected to different angiogenic stimuli; in vivo by intravitreal administration to mice pups in a model of oxygen-induced retinopathy (OIR), and to adult mice after choroid neovascular lesions induced by laser photocoagulation. Morphological, functional and immunohistochemical analyses were performed to show the efficacy of treatment.

Results: Uparant treatment was highly effective in reducing both vascular sprouting in vitro and neoangiogenesis in vivo. In addition, UPARANT decreased vessel leakage and improved visual function in OIR mice.

Conclusions: Uparant appears to be a very promising molecule to counteract neoangiogenesis promoted by different pathological conditions.

 

 

 

945

--------------------------------------------------------------------------------------------

Glaucoma

New useful thoughts about old ideas; or "It is better to be certain than to guess"

 George L. Spaeth,  George Spaeth
Glaucoma Department, Wills Eye Hospital, USA

1) Means and standard deviations are not reliable indicators of clinical significance. Pay attention to what is ALWAYS certainly pathologic, such as a notched disc; risk calculators are poor guides to which individuals need treatment. A finding that is not in the range of ALWAYS pathologic is usually of unknown value. 2) Cup/disc ratios are not valid descriptors of nerve damage and are poorf diagnostic guides. They do not take into account disc size or cup eccentricity (rim width), and they correlate poorly with field loss. There is no c/d ratio which is ALWAYS certainly diagnostic of glaucoma. Learn the Disc Damage Likelihood Scale (DDLS); it is a valid, relevant indicator of stage of optic nerve damage, which must be known with certainty for treatment to be appropriate.3) Bias is insidious and powerful. Learn to recognize personal, institutional and professional biases such as the incorrect belief that "Objective" findings are more valuable than "Subjective" findings, and the incorrect advice to look at the field before examining the disc. 4)"Hypotony" is often a misleading word. The threshhold intraocular pressure below which problems occur can not be accurately predicted. Just as what level of elevated IOP will cause field loss in an individual can not be certainly predicted, so also the pressure below which the level of pressure will cause problems can not be accurately predicted with certainty. 5) The evidence that trabeculectomy itself causes cataracts is unconvincing. Postoperative cataract usually results from surgical trauma, or use of topical corticosteroids for more than 4 weeks.

 

 

 

948

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Innovation in Ophthalmology

CYTOMEGALOVIRUS RETINITIS IN HIV-NEGATIVE PATIENTS, RETRISPECTIVE INTERVENTIONAL CASE SERIES AND PRACTIVCAL APPROACH TO PATIENTS MANAGEMENT

 

 Vicktoria (Vicky) Vishnevskia-Dai1,  Yinon Shapira1,2,  Joseph Moisseiev1,  Galia Rhav3,  Avichai Shimoni4,  Raz Somech5
1Ophthalmology department the Goldschleger eye institute Sheba Medical Center Tel Aviv University, Ocular oncology and Inflammatory eye diseases service, Israel
2Ophthalmology, The Ruth and Bruce Rappaport Rambam Faculty of Medicine technion, Israel
3Infectious Diseases Unit, Sheba Medical Center Tel Aviv University, Israel
4The Bone Marrow Transplantation Department, Sheba Medical Center Tel Aviv University, Israel
5Pediatric Department B, Edmond and Lily Safra Children's Hospital, and Cancer Research Center,Pediatric Immunology Service, Jeffrey Modell Foundation (JMF) Center, Sheba Medical Center Tel Aviv University, Israel

Purpose: Characterization of the CMV retinitis in HIV-negative patients, review the management strategy and treatment results

 

Methods: Retrospective case series of CNV retinitis in HIV-negative patients treated in our center between the years 2007 and 2013.

CMV retinitis was defined by characteristic ophthalmoscopic findings and was confirmed by PCR and/or CMV viremia.

The patients received systemic or intravitreal , single or dual- antiviral agent in accordance with the ocular presentation and baseline disease.

Demographic and medical characteristics as well as the features of CMV retinitis and response to treatment were evaluated.

 

Results: The series comprises of 9 patients in the average age of 27y. The follow up was in average 12 months.

One patient exhibited a rapid response to a single systemic antiviral agent (ganciclovir). All other patients were administered a combined intravenous and/or intravitreal dual-anti viral agent. Following full treatment all patients showed improvement of the CMV retinitis as well as resolution of the viremia. Final patient outcome was evidently related to the underlying systemic morbidity.

 

Conclusions: Our experience may suggest an aggressive treatment approach in the non-HIV related severe CMV retinitis cases. Patient management should be tailored in a multidisciplinary approach in accordance with the disease severity, ocular presentation , baseline disease, patient age, , systemic condition and response to treatment.

 

 

 

1019

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Basic Science

Regulated expression of recombinant anti-VEGF single chain antibody fragments – towards personalized medicine in neovascular retinal disorders

 Tobias Wimmer,  Birgit Lorenz,  Knut Stieger
Department of Ophthalmology, Justus-Liebig-University Giessen, Germany

Purpose: Most retinal neovascular disorders are associated with up-regulation of the vascular endothelial growth factor (VEGF) expression. Currently, disorders like age-related macular degeneration, diabetic retinopathy and retinal vein occlusion are treated with repeated injections of anti-VEGF molecules like Bevacizumab, Ranibizumab, or Aflibercept. Repeated injections of anti-VEGF molecules can be connected to severe side effects and represent a financial burden to the patients. The aim of this project is to develop an alternative strategy i.e. controlled expression of anti-VEGF molecules within the retina.

 

Methods: The open reading frames of ranibizumab were cloned into an expression plasmid separated by an internal ribosomal entry site (IRES) (Ra01). The construct was mutated to generate ranibizumab single chain variable fragments (Ra02-Ra05). Expression was verified by Western blotting. Biological activity, VEGF binding properties and the doxycycline depend induction of anti-VEGF expression was tested. An AAV2/5 vector was generated containing the optimal variant Ra02.

 

Results: Ra variants 01-05 were detected in cell culture medium. While VEGF binding affinity was significantly lower compared to Lucentis®, the inhibition of cell migration was comparable and the maximum inhibition of Ra01 and Ra02 was reached at lower doses. The expression of Ra02 was shown to be controllable with the TetOn-system® as plasmid and AAV vector construct.

 

Conclusion: Anti-VEGF molecules related to ranibizumab can be produced in eukaryotic cells after AAV mediated gene transfer in a controlled manner in vitro and display comparable biological activity as Lucentis®. These results are the basis for a gene-based therapy in human VEGF overexpressing mice, a model for human neovascular disorders.

 

 

 

714

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Drug Development

Enhanced corneal permeation of coumarin-6 using nanoliposomes containing dipotassium glycyrrhizinate: in vitro mechanism and in vivo permeation evaluation

 Xianggen WU
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of medical Sciences, China

The elasticity of the lipid bilayer of nanoliposomes regarding in vitro cellular uptake/mechanics and in vivo corneal permeation through ocular topical routes was investigated. Flexible nanoliposomes, using dipotassium glycyrrhizinate as edge activator, and their physical properties, membrane elasticity, cellular uptake characterizations and mechanisms, as well as in vivo corneal permeation using rabbits and mice as experimental animals, were investigated and compared with the conventional liposomal formulation composed of soybean phosphatidylcholine and cholesterol. Flexible nanoliposomes required less energy to prepare and had elastic lipid membranes, and the elasticity was confirmed by the transmission electron microscopy and elasticity evaluation. Compared with nanoliposomes, flexible nanoliposomes showed significantly higher cellular uptake of coumarin-6 in human corneal epithelial cell line. Moreover and interestingly, the flexible nanoliposomes showed different cellular uptake mechanisms in cells. The uptake of nanoliposomes occurred via an energy-dependent process, while it was mainly an energy-independent process to the flexible nanoliposomes. Flexible nanoliposomes also showed significantly higher corneal penetrating ability in in vivo testing. The coumarin-6 levels of the flexible liposomal formulation in the mouse cornea were 107.31%, 228.26%, and 136.50% higher than those of the liposomal formulation at the 30, 60 and 120 min time points, respectively, after four instillations to the mice eyes, and were 39.64%, 172.09%, and 103.27% higher when it was tested in rabbits. Therefore, the fluidity of the liposomal membrane differently affected cellular uptake/internalization and in vivo corneal penetration of the nanoliposomes, and flexible nanoliposomes might be a promising therapeutic tool for the treatment of ocular surface disorders.

 

 

900

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Drug Development

Fluorescein uptake in the anterior ocular segment to test a new topical drug delivery device for ocular medication

 Rene Wubbels1,  Wilbur de Kruijf2,  Jeroen van Rooij1
1ROI, Rotterdam Eye Hospital, Netherlands
2Device development, Medspray BV, Netherlands

Purpose: To compare fluorescein uptake in the anterior eye: conventional eye drops versus a newly developed spray device.

Methods: In this `proof of principle` study, 18 healthy volunteers participated. A fluorescein solution (20 mg/mL) was administered to one eye as a conventional drop (50 µL), and as an ocular mist (10 µL) to the other of all subjects. Autofluorescence (photons/s) was repeatedly measured up to 100 min after application in the cornea, the anterior chamber (AC) and the lens. Area Under the Curve (AUC) was calculated by means of a linear trapezoidal rule approximation.

Results: AUC (mean ± SD) for the cornea was (363 ± 431)*104 photons after drop and (154 ± 265)*104 photons after mist application (p = 0.005). For the AC these values were (6.9 ± 10.3)*104 and (2.9 ± 5.4)*104 photons respectively (p = 0.14). Within subjects, bilateral correlation is strong (cornea: R2 = 0.63). The average AUC (drops/spray) ratio was 4.1 ± 3.4 for the cornea and 4.4 ± 5.8 for the AC. Autofluorescence data obtained in the lens did not allow reliable AUC calculations.

Conclusion: The intra-ocular penetration of fluorescein applied as a mist has been demonstrated. As the amounts getting to the ocular surface are not equivalent for both delivery methods, no conclusions are drawn on relative bioavailability. Still, a substantial reduction of the volume of topical, ocular administered substances appears to be feasible.