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ARVO 2016 Annual Meeting Abstracts 374 Ocular Surface Health and Disease Tuesday, May 03, 2016 3:45 PM–5:30 PM Exhibit/Poster Hall Poster Session Program #/Board # Range: 3853–3896/A0001–A0044 Organizing Section: Cornea Program Number: 3853 Poster Board Number: A0001 Presentation Time: 3:45 PM–5:30 PM APR-246/PRIMA-1Met inhibits and reverses squamous metaplasia in human conjunctival epithelium Cheng Li, Wei Li, ZUGUO LIU, JIng Li. Ophthalmology and Visual Sciences, Eye Inst & Affiliated Xiamen Eye Ctr, Xiamen, China. Purpose: Squamous metaplasia is a common pathological condition in ocular surface diseasesfor which there is no therapeutic medication in clinic. In this study, we investigated the effect of a small molecule APR-246/PRIMA-1Met on squamous metaplasia in human conjunctival epithelium. Methods: Human conjunctival explants were cultured for up to 12 days under airlift conditions. Epithelial cell differentiation and proliferation were assessed based on K10, K14, K19, Pax6, MUC5AC, and p63 immunostaining patterns. β-catenin and TCF-4 immunofluorescent staining and real-time PCR characterized Wnt signaling pathway involvement. Pterygium clinical samples were cultured under airlift conditions with or without APR-246 for 4 days. p63, K10, β-catenin and TCF-4 expression in pterygial epithelium was determined by immunofluorescent staining and real-time PCR. Results: Airlifting conjunctival explants resulted in increased stratification and intrastromal epithelial invagination. Such pathology was accompanied by increases in K10, K14 and p63 expression whereas K19 and Pax6 levels declined compared to those in freshly isolated tissue. On the other hand, APR-246 reversed all of these declines in K10, K14, and p63 expression. Furthermore, K19 and Pax6 increased along with rises in goblet cell density. These effects of APR-246 were accompanied by near restoration of normal conjunctival epithelial histology. APR-246 also reversed squamous metaplasia in pterygial epithelium that had developed after 4 days ex vivo culture. Conclusions: Reductions in squamous metaplasia induced by APR246 suggest it may provide a novel therapeutic approach in different squamous metaplasia associated ocular surface diseases. Commercial Relationships: Cheng Li; Wei Li, None; ZUGUO LIU, None; JIng Li, None Program Number: 3854 Poster Board Number: A0002 Presentation Time: 3:45 PM–5:30 PM The Effect of Betadine on Vision Caren Oquindo1, William H. Ridder1, Kavita Dhamdhere2, James A. Burke2. 1Southern California College of Optometry, Marshall B. Ketchum University, Fullerton, CA; 2Allergan, Inc., Irvine, CA. Purpose: Betadine is instilled topically on the eye prior to ocular surgical procedures to decrease the risk of endophthalmitis. Its effects on functional vision and the cornea have not been determined. The purpose of this study was to determine the effects of 5% betadine on vision function, corneal integrity and subjective complaints. Methods: Twenty subjects were chosen to participate in this study (Ten young: 25.8 +/- 2.94; and ten old: 58.2 +/- 5.59). LogMAR acuity, contrast sensitivity, corneal fluorescein staining, and the Schein dry eye questionnaire were measured before and after 60 µl of 5% betadine was applied to one eye, randomly chosen, (baseline, 5, 30, and 60 minutes and 4 and 24 hours post-application). Contrast sensitivity at 14 cpd was determined with a spatial two-alternative, forced choice procedure (BeethovenTM software). The NEI grid was used to grade corneal staining. Subjective complaints were monitored using the Schein dry eye questionnaire. Results: The data were analyzed with an ANOVA (linear mixedeffects model). The logMAR acuity was significantly reduced from baseline at the 30 and 60 minute visits (all p values < 0.05) and contrast sensitivity was reduced from baseline at 5, 30, and 60 minutes after betadine application (all p values < 0.0001). Total corneal staining, maximum NEI sector staining, and the Schein dry eye questionnaire were significantly different from baseline at every visit (all p values < 0.05). Conclusions: There was an increase in corneal staining and subjective complaints that lasted 24 hours and a decrease in functional vision that lasted 1 hour after 5% betadine instillation. The central corneal staining cleared more rapidly than the peripheral cornea; therefore, visual acuity recovered more quickly than overall corneal staining and subjective complaints. This may have resulted from pooling of Betadine at the limbus and lid margins. 5% Betadine application significantly affects corneal integrity which decreases functional vision and increases subjective complaints. Commercial Relationships: Caren Oquindo, Allergan, Inc. (F); William H. Ridder, Allergan, Inc. (F); Kavita Dhamdhere, Allergan, Inc.; James A. Burke, Allergan, Inc. Support: Allergan Inc Program Number: 3855 Poster Board Number: A0003 Presentation Time: 3:45 PM–5:30 PM The Effect of Betadine on Vision and on Cornea in Rabbits Kavita Dhamdhere, Alexandra S. Almazan, Michael Engles, James A. Burke. Allergan Inc, Irvine, CA. Purpose: Betadine is instilled topically on the eye prior to most ocular surgical procedures to decrease the risk of endophthalmitis. Its effects on vision and the cornea have not been determined. The purpose of this study is to evaluate the effects of Betadine® 5% sterile ophthalmic prep solution (Alcon, Fort Worth, TX) on vision (function) and corneal integrity (structure) in rabbits. Methods: Twenty-four female Dutch-Belted rabbits with normal vision were included in this study (Mean age: 2.5 ± 0.94 yrs), divided in two groups: vision group (n=18) and structure group (n=6). The study eye in each group was randomized and was treated with a drop of tetracaine hydrochloride ophthalmic solution USP 0.5% (Bausch and Lomb, Tampa, FL) for topical anesthesia, then with 50 μl of Betadine. Betadine was washed out with sterile saline after 2 mins of exposure. Monocular visual acuities (VA) and corneal fluorescein staining (CFS) were measured at baseline and at 1, 4 and 24 hours post treatment in conscious rabbits. OptoMotor™ system (Prusky et al, 2004; Douglas et al, 2005) was used to collect VA (spatial thresholds) and the total number of punctate stains were used to grade CFS. The effect of Betadine on vision and cornea was evaluated as a function of change from the baseline. The data is presented in the form of mean ± SD. Results: The data were analyzed with an ANOVA for multiple comparisons. Mean baseline VA and CFS was respectively 0.789 ± 0.017 cpd and 3 ± 0.8 total punctate stains in the study eye. The VA was significantly reduced post betadine dosing at each follow up time point (0.470 ± 0.17, 0.511 ± 0.17 and 0.609 ± 0.19 respectively at 1, 4 and 24 hrs; p < 0.0001). Total CFS was significantly increased from baseline at each follow up time point (80 ± 4.4, 72 ± 10 and 43 ± 5.1 respectively at 1, 4 and 24 hrs; p < 0.0001). Two thirds of the animals had conjunctival hyperemia and/or blepharospasm up to 24 hrs post Betadine application. Conclusions: There was a decrease in VA and an increase in corneal staining that lasted for 24 hours after 5% betadine instillation. The central corneal staining cleared more rapidly than the peripheral These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts cornea. This may have resulted from pooling of Betadine at the limbus and lid margins. Commercial Relationships: Kavita Dhamdhere; Alexandra S. Almazan, None; Michael Engles, None; James A. Burke, None Program Number: 3856 Poster Board Number: A0004 Presentation Time: 3:45 PM–5:30 PM Hoxc8 misexpression prevents embryonic eyelid fusion and transforms cornea and conjunctiva into keratinized skin Lara S. Carroll. Moran Eye Center, University of Utah, Salt Lake City, UT. Purpose: Hox gene expression is excluded from the developing eye, although several members of this gene family are expressed in fetal and adult epidermis. We found that misexpression of Hoxc8 throughout mouse embryonic ectoderm results in pups born with their eyes open (eyes open at birth-EOB), and the cornea and conjunctiva transformed into a skin-like, keratinized epidermis. We examined expression of essential genes for eyelid fusion and corneal and conjunctival development to determine how Hoxc8 expression hijacks these specific epithelial programs to initiate epidermal development. Methods: Mice carrying a conditional CAGGSHoxc8iresEGFP cassette were bred to Wnt6cre mice to create double heterozygous offspring that misexpress Hoxc8 throughout the Wnt6 domain, which is expressed broadly across the embryonic ectoderm. Pregnant dams were sacrificed to collect offspring between E11.5 and birth for phenotypic characterization. Tissue was collected for RNA and immunohistochemical analysis of genes involved in eyelid fusion pathways and corneal differentiation, including: Wnt, Bmp, Shh, Fgf10, ERK, and retinoic acid signaling, as well as Pax6 and various cytokeratin markers. Results: Newborn H&E staining showed sloughing squamous epithelium covering the cornea and conjunctiva of conditional mutants, with aberrant hair follicles present in the conjunctiva. Examination of earlier timepoints showed that mutants fail to develop an eyelid fusion front with associated migrating periderm and aligned F-actin fibers. Mutant eyelid tips lacked localized expression of p-cJun, p-ERK, Shh, Bmp4, and failed to downregulate Wnt signaling in contrast to age-matched control embryos. Conversely, mutant corneal epithelium failed to upregulate Wnt signaling, as evidenced by the lack of Lef1. Pax6, which is essential for maintaining corneal epithelium, was absent in mutant corneas, while skin specific keratins, K1 and K10 were upregulated throughout the corneal and conjunctival epithelium. Conclusions: Emerging evidence suggests that Hox genes play a key role in regionalization of the skin and epidermal organs. Aberrant expression of Hoxc8 in the developing eye field ectoderm aborts Pax6 expression in corneal epithelium, leading to corneal and conjunctival keratinization, and disrupts most known signaling pathways for eyelid fusion, the last major morphogenetic event to occur in mammalian eye development. Commercial Relationships: Lara S. Carroll Support: 2RO1EY017182 Program Number: 3857 Poster Board Number: A0005 Presentation Time: 3:45 PM–5:30 PM Identification and Characterization of Long Noncoding RNA Contributions to Mouse Corneal Development Weiwei Chen1, 2, Shuai Yang1, 2, Zhonglou Zhou1, 2, Xiaoting Zhao1, 2, Dongsheng Yan1, 2. 1State Key Laboratory of Ophthalmology, Wenzhou Medical University, Wenzhou, China; 2School of Ophthalmology and Optometry, Wenzhou Medical University, Wenzhou, China. Purpose: Long noncoding RNAs (lncRNAs) are important regulators of cellular functions. However an extensive in-depth analysis of their expression profile and function in mouse corneal development is still lacking. Here we investigated lncRNAs profiles of the developing mouse cornea by microarray and bioinformatics. Methods: Affymetrix mouse transcriptome 1.0 assay identified corneal lncRNA profile changes occurring between birth and 8 weeks in mice. Real-time RT-PCR analysis validated array findings. Gene ontology (GO) and KEGG pathway mapping of protein-coding genes adjacent to signature lncRNAs clarified potential lncRNA roles in regulating corneal development. Results: In newborn and 8-week-old mice, 41,987 protein-coding and noncoding gene transcripts were identified. In these two subsets, 19, 623 of this ensemble are lncRNAs annotated in public data banks. During development, 1,272 lncRNAs underwent ≧ two-fold changes in expression levels. qPCR analysis confirmed gene microarray analysis results since there was 90% agreement between the two procedures in identifying lncRNAs contributing to this process. GO analysis of protein-coding genes proximal to lncRNA signatures resolved numerous neighboring protein coding genes regulating cell division and adhesion as well as collagen and cytokine production, suggesting a role for signature lincRNAs in controlling corneal development. Conclusions: Time dependent changes in lncRNA expression patterns occurring during mouse corneal development suggest that they play an important role in regulating this process. Commercial Relationships: Weiwei Chen, None; Shuai Yang, None; Zhonglou Zhou, None; Xiaoting Zhao, None; Dongsheng Yan, None Support: 973 Project (2012CB722303) from the Ministry of Science and Technology of China, and Science Foundation of Wenzhou Medical University QTJ11020 Program Number: 3858 Poster Board Number: A0006 Presentation Time: 3:45 PM–5:30 PM IKZF1 Expression and Upregulation by polyI:C in Human Ocular Surface Epithelial Cells Mayumi Ueta1, Hiromi Nishigaki1, Katsuhiko Shinomiya2, Norihiko Yokoi2, Chie Sotozono2, Shigeru Kinoshita1. 1Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan; 2Department of Opthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan. Purpose: Stevens-Johnson syndrome (SJS) / toxic epidermal necrolysis (TEN) are acute inflammatory vesiculobullous reactions of the skin and mucosa, such as the ocular surface and oral cavity. In genome wide association study (GWAS), we found the association between IKAROS Family Zinc Finger 1 (IKZF1) and Cold medicine related SJS/TEN with severe ocular complication (CM-SJS/TEN with SOC). Ikaros is a transcription factor which regulates numerous biological events, but the function of it was reported only in the immune cells such as lymphocytes. On the other hands, we have suggested that epithelium might be contribute to the pathobiology of CM-SJS/TEN with SOC, because EP3, the protein of PTGER3, These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts which SNPs significantly associated with CM-SJS/TEN with SOC, was dominantly expressed on the ocular surface epithelium and EP3 on the epithelium negative regulated ocular surface inflammation. Furthermore, we also reported that TLR3, which SNPs significantly associated with CM-SJS/TEN with SOC in Japanese, also strongly expressed on the ocular surface epithelium and TLR3 positively regulated ocular surface inflammation. In this study, we examined the expression of IKZF1 in the human ocular surface (conjunctival and corneal) epithelial cells, and investigated whether it could be upregulated by TLR3 ligand. Methods: For quantitative RT-PCR, primary human conjunctival epithelial cells were harvested from conjunctival tissue obtained at conjunctivochalasis surgery and cultured using the previously described method. Primary human corneal cells were harvested from the rest of the limbus of import donor cornea tissue after using for corneal transplantation. For RT-PCR and immunohistology, conjunctival tissue obtained at conjunctivochalasis surgery was used. Results: We found that human conjunctival epithelium expressed IKZF1 mRNA by RT-PCR and using quantitative RT-PCR, we found that it could be upregulated by TLR3 ligand, polyI:C. We also found that IKZF1 mRNA expression in corneal epithelial cells could be upregulated by polyI:C. Immunohistology showed that IKZF1 protein also expressed in conjunctival epithelium as same as CD3 positive T cells. Conclusions: Our results might suggest that IKZF1 in the ocular surface could contribute to the ocular surface inflammation in the SJS/TEN. Commercial Relationships: Mayumi Ueta, None; Hiromi Nishigaki, None; Katsuhiko Shinomiya, None; Norihiko Yokoi, None; Chie Sotozono, None; Shigeru Kinoshita, None Support: This work was supported by grants-in-aid from the Ministry of Education, Culture, Sports, Science and Technology of the Japanese government (BioBank Japan Project), and by the JSPS Core-to-Core Program, A. Advanced Research Networks and also partly supported by grants-in-aids for scientific research from the Japanese Ministry of Health, Labor and Welfare, and a research grant from the Kyoto Foundation for the Promotion of Medical Science and the Intramural Research Fund of Kyoto Prefectural University of Medicine. Program Number: 3859 Poster Board Number: A0007 Presentation Time: 3:45 PM–5:30 PM Ocular Surface Adverse Events of Systemic Inhibitors of the Epidermal Growth Factor Receptor (EGFRi): a Prospective Trial Alejandro Saint-Jean1, 2, Ana Eixarch2, Noemi Reguart4, Nuria Pardo4, Cristina Castella2, 3, Maite Sainz De La Maza2, Alfredo Adan Civera2, Bernardo Sanchez Dalmau2, Marta Aldea5, Ruben Torres2. 1Ophthalmology, CHU Nantes, Nantes, France; 2 Ophthalmology, Hospital Clinic Barcelona, Barcelona, Spain; 3 Clinique Sourdille, Nantes, France; 4Oncology Department, Hospital Clinic, Barcelona, Spain; 5Unit of Infections and Cancer (UNIC), Cancer Epidemiology Research Programme, Institute of Oncology, IDIBELL, Barcelona, Spain. Purpose: Controversy exists regarding the safety of agents that systemically inhibits epidermal growth factor receptor (EGFRi) in the ocular surface for oncologic patients. We performed a prospective observational clinical study to compare the ocular surface toxicity of systemic EGFRi between a case group and a control group without EGFRi. Methods: Patients with lung or colon cancer (34 lung and 18 colorectal adenocarcinoma) were divided in two groups: 26 patients treated with systemic EGFRi wether erlotinib for lung cancer or panitumumab for colon cancer (18 lung and 8 colon cancer, mean age 73.4+/-7.3 years old) and 26 patients in a control group without EGFRi treatment matched by the stage of the disease (16 lung and 10 colon cancer, mean age 63.5+/-8.6 years old). Patients in both groups patients were chemotherapy naive (de novo treatment). Four visits were scheduled in a one year period comparing signs and symptoms in terms of symptom questionnaires (SIDEQ, OSDI and AVS), corneal fluorescein staining (Oxford test), tear production (Shirmer’s test), and a quantitative evaluation of conjunctival chemosis and hyperemia. Basal epithelial cell density (BECD) and corneal subepithelial nerve fiber density (SNFD) were measured and compared using confocal microscopy (Heidelberg Ingeneering, Germany). The differences in each variable between the 2 groups were compared with the analysis of variance (ANOVA) (quantitative variables). P value <0.05 was considered significant for all comparisons. Results: Statistically significant differences were found between patients under EGFRi and the age-matched controls in terms of conjunctival chemosis and Oxford test in all visits (P< 0.05). When cases and controls were evaluated separatedly, the case group showed a significant worse evolution in symptoms (SIDEQ and OSDI), Oxford test and Shirmer’s test (all P <0.05). However the control group did not show significant differences among visits. We did not find significant differences in terms of ECD and SNFD between cases and age/type of cancer matched controls. Conclusions: Systemic EGFRi may increase conjunctival chemosis and corneal fluorescein staining. Oncologic patients treated with EGFR should be monitored closely with ophthalmologic examinations to detect dry eye Commercial Relationships: Alejandro Saint-Jean, None; Ana Eixarch; Noemi Reguart, None; Nuria Pardo, None; Cristina Castella, None; Maite Sainz De La Maza, None; Alfredo Adan Civera, None; Bernardo Sanchez Dalmau, None; Marta Aldea, None; Ruben Torres, None Clinical Trial: ASJ-PAN-2012-01 Program Number: 3860 Poster Board Number: A0008 Presentation Time: 3:45 PM–5:30 PM A role of primary cilia in Anterior Segment Dysgenesis disorders Carlo Iomini, Grisanti Laura, Ekaterina Revenkova. Ophthalmology, Icahn School of Medicine at Mount Sinai, New York, NY. Purpose: The development of the anterior segment (AS) of the mammalian eye involves tightly coordinated cellular interactions between tissues of different embryonic origins. Defects in this process lead to severe ocular disorders including aniridia, microcornea and primary congenital glaucoma of unclear pathophysiology. Interestingly, dysgenesis of the AS was reported in patients affected by ciliopathies. Here we investigate the role of cilia during development of the AS in mice. Methods: Phenotypic analysis of the AS was performed using light and transmission electron microscopy. Expression of ciliary-mediated pathways target genes or transcription factors implicated in AS morphogenesis were assessed using qRT-PCR and immunostaining approaches. Cell proliferation of periocular mesenchyme was assessed by BrdU incorporation and immunostaining. Results: Null mutations that prevent cilia assembly such as Ift88-/are lethal at E9.5-10.5. In Ift88-/- embryos the optic vesicle, although defective, is present. In contrast, at E10.5 the optic cup do not form as observed in wild-type embryos. To overcome mid-gestation lethality generated a conditional KO mouse Wnt1-Cre;IFT88flox/(Ift88 cKO) in which the Ift88 gene is excised in all migrating mesenchymal cells of neural crest origin. cKO mice display strong craniofacial defects and die at postnatal day P0. Early eye patterning These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts appears normal. However, later in development cKO mice display significant reduction of the anterior chamber with occasional fusion of the endothelium to the lens epithelium, a thinner stromal ECM and corneal endothelial malformations. Interestingly, although migration of neural crest-derived cells appears normal, we detected reduction of mesenchymal cells at the angle between the stroma and the optic cup at E17.5. Gene expression analysis revealed reduction of Hedgehog pathway target genes and Ptx2 but not Foxc1, both encoding for transcription factors linked to the Axenfeld-Rieger Syndrome. Immunofluorescence analysis confirmed reduction of PTX2 expression at the iridocorneal angle and detected mislocalization of cells expressing FOXC1. Conclusions: We have uncovered a pivotal role of primary cilia in the development of the AS and highlighted their involvement in the etiology of ocular disorders derived from AS dysgenesis. We have also provided an accessible paradigm to address the role of primary cilia in complex tissue morphogenesis. Commercial Relationships: Carlo Iomini, None; Grisanti Laura, None; Ekaterina Revenkova, None Support: NIH Grant EY022639 and RPB Dolly and Unrestricted Awards Program Number: 3861 Poster Board Number: A0009 Presentation Time: 3:45 PM–5:30 PM PROSE lens use for exposure keratopathy in trigeminal and facial nerve palsies Elizabeth Marlow, Jessica Ciralsky, Michelle Lee, Gary Lelli. Ophthalmology, Weill Cornell Medical College, New York, NY. Purpose: For patients with severe ocular surface disease, prosthetic replacement of the ocular surface ecosystem (PROSE) devices have shown efficacy in improving visual acuity and the ocular surface. Individuals with facial and trigeminal nerve palsies are at particularly high risk for exposure keratopathy, making PROSE a potentially beneficial tool in the management of these cases. This retrospective, observational clinical study reviewed the effects of initiating PROSE wear on nine eyes affected by facial (VII), trigeminal (V1), or both (VII and V1) cranial nerve palsies. Methods: Best corrected visual acuity (BCVA) at the initial PROSE fitting, three months after initiating PROSE wear, and BCVA achieved at any time after starting PROSE wear were compared. In addition, the quality of the ocular surface before starting PROSE and at the time of longest PROSE wear was compared. Results: After three months of PROSE wear, patients gained an average of 0.9 ± 1.4 lines on the Snellen eye chart compared to initial BCVA. The average maximal gain in lines on the Snellen eye chart achieved at any time after starting PROSE was 1.9 ± 1.3 lines, which occurred an average of 5.3 ± 5.4 months after starting PROSE wear. The ocular surface quality improved in 8 out of 9 eyes with the resolution of punctate epithelial erosions (n = 7), reduced corneal haze (n = 2), and reduced mucus strands (n = 2) being the most commonly sited changes. Conclusions: These findings show that the PROSE device can be an effective tool in the management of exposure keratopathy secondary to trigeminal (V1) and facial nerve palsies. Commercial Relationships: Elizabeth Marlow, None; Jessica Ciralsky, None; Michelle Lee, None; Gary Lelli, None Support: Unrestricted support from Research to Prevent Blindness Program Number: 3862 Poster Board Number: A0010 Presentation Time: 3:45 PM–5:30 PM Impact of conjunctival autograft on pterygium treatment: evaluation of related symptoms and patients satisfaction after surgery Bruna Duarte Moron de Andrade, Giovana Capecci Siqueira, Paulo Dechichi Neto, Augusto Terra Baccega, Marina Gonçalves Monteiro Viturino, Ana Luiza Mylla Boso, Daniella de Paiva Almeida, Monica Alves. Department of Ophthalmology, UNICAMP, Campinas, Brazil. Purpose: Pterygium is a fibrovascular condition of the ocular surface that can cause a broad range of irritative and visual symptoms. Controversy exists regarding the pterygium mechanisms, management, surgical techniques, adjuvant approaches and its impact on patients’ quality of life. We performed a retrospective survey to better understand the impact of pterygium related symptoms before surgery and patients satisfaction after excision surgery followed by conjunctival autograft transplantation with fibrin glue. Methods: All patients underwent surgery that consisted of an extensive removal of the pterygium fibrovascular tissue, followed by an autologous conjunctival graft fixed with fibrin glue to cover the bare scleral area, performed by the same surgeon. A total of 215 patients were contacted by phone call and asked to answer a simple 2 questions survey. First, a graduation of the symptoms related to pterygium before surgical intervention, such as pain, irritation, tearing, red eye, photophobia, burning, body sensation, scaled from 0-10 (0 asymptomatic and 10 very important symptoms). We then classified as mild (0-3), moderate (4-7) and severe (8-10). Then, the patients were asked about their satisfaction on surgery results, scaled from 0-10 (ranging from dissatisfied to fully satisfied). Results: Patients mean age was 41.1 (min-19/max-82) years old and the mean of days after surgery was 967.2 days (min-347/max-2155). Symptoms were referred as severe (71.8%), moderate (23.9%) and mild (4.3%). After surgery most patients were fully satisfied and the mean grade was 9.4; 0.9% (0-3), 1.9% (4-7) and 97.1% (8-10). Conclusions: The present study shows that pterygium excision surgery using conjunctival autograft transplantation and fibrin glue improved quality of life related to pterygium symptoms on patients affected by the condition with high rates of satisfaction. However, there is still lack of understanding on pterygium mechanisms, no consensus on best surgical technique and the recurrence rates remain a challenge, justifying the need of further research on this area. Commercial Relationships: Bruna Duarte Moron de Andrade, None; Giovana Capecci Siqueira, None; Paulo Dechichi Neto, None; Augusto Terra Baccega, None; Marina Gonçalves Monteiro Viturino; Ana Luiza Mylla Boso, None; Daniella de Paiva Almeida, None; Monica Alves, None Support: Fapesp grant #2014/19138-5 Program Number: 3863 Poster Board Number: A0011 Presentation Time: 3:45 PM–5:30 PM Correlation between Meibomian Gland Dysfunction and Keratitis in Patients with Demodex Brevis Infestation Lingyi Liang1, Hongmin Ke1, Scheffer C. Tseng2. 1Cornea, Zhongshan Ophthalmic Center, Guangzhou, China; 2Ocular Surface Research and Education Center, Miami, FL. Purpose: To investigate whether there is any correlation between meibomian gland dysfunction (MGD) and keratitis in patients with ocular demodicosis. Methods: Prospective, cross sectional, comparative and controlled case series. Sixty study patients with ocular demodicosis and 45 gender- and age- matched dry eye patients as the control without demodicosis. All patients were younger than 35 years-old. Diagnosis These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts of demodicosis was based on microscopic counting of D. folliculorum and D. brevis of epilated lashes. Severity of keratitis and MGD was graded by photography and meibography, respectively, in a masked fashion. Statistical correlation between Demodex infestation, MGD and keratitis were analyzed. Results: MGD defined by meibography and meiboscore was significantly more prevalent and more severe in the study group than the control group (both P<0.05). Meibomian gland loss was worse in the upper lid in study patients but similar in upper and lower lids in control patients. Superficial punctate keratitis was found in both groups, while limbitis, stromal infiltration, vascularization, scarring, and perforation were only noted in study patients. Both univariate and multivariate analyses showed that the meiboscore was significantly correlated with higher D. brevis count and more severe keratitis, but not with age, gender, history course, and D. folliculorum count. Bayesian network models tested by R package bnlearn suggested that D. brevis counts contributed to both keratitis and MGD. Rapid resolution of keratitis and blepharoconjunctivitis was accompanied by significant reduction of the demodex count in 48 patients receiving lid scrub using tea tree oil, which had been reported to kill Demodex mites. Conclusions: Ocular demodicosis, especially by D. brevis, may play a causative role in MGD and keratitis in young patients. Its potential pathogenic role in MGD in the older population cannot be ignored. Commercial Relationships: Lingyi Liang, None; Hongmin Ke, None; Scheffer C. Tseng, Biotissue (P) Support: National Eye Institute, the National Institutes of Health (R44 EY019586); National Natural Science Foundation of China(81300739); Ministry of Education of China (20110171120104); Technological Project Foundation of Guangdong Province(2011B031800274, 2012B031800456) Program Number: 3864 Poster Board Number: A0012 Presentation Time: 3:45 PM–5:30 PM The Evaluation of Worldwide Distribution of Adenoviral Genotypes in Acute/Epidemic Keratoconjunctivitis and Adenoviral-negative Keratoconjunctivitis with Next Generation Sequencing Cecilia S. Lee1, Aaron Y. Lee1, Lakshmi Akileswaran1, David W. Stroman2, Kathryn Najafi-Tagol2, Steve Kleiboeker4, Anna Wald3, Russell N. Van Gelder1. 1Ophthalmology, University of Washington, Seattle, WA; 2NovaBay Pharmaceuticals, Inc., Emeryville, CA; 3Infectious Diseases, University of Washington, Seattle, WA; 4ViraCor-IBT Laboratories, Inc, Lee's Summit, MO. Purpose: Conjunctivitis is among the most common outpatient conditions and causes significant ocular morbidity worldwide. The most severe and common form of viral conjunctivitis is epidemic keratoconjunctivitis (EKC), caused by certain strains of adenovirus (Ad). However, the geographic distribution of other Ad genotypes and the role of the ocular surface microbiome (OSM) in EKC have not been established. We investigated the worldwide distribution of Ad genotypes and the OSM associated with keratoconjunctivitis (KC). Methods: A variety of analyses were conducted on samples obtained in a large international clinical trial (NV-422 Phase IIB, NovaBay Pharmaceuticals, Inc). The inclusion criteria were clinical signs and symptoms consistent with KC and evidence of Ad using the rapid screening test (Rapid Pathogen Screening Inc., Saratosa, FL). Bilateral conjunctival swabs were obtained on day 1, 3, 6, 11, and 18. Ad genotype was determined by sequencing of Ad hexon gene (ViraCor-IBT Laboratories, Inc, Lee’s Summit, MO) and the OSM was assessed using next generation sequencing (NGS) on selected samples. The difference in the distribution of Ad genotypes was assessed with Fisher exact. Results: Overall, 5000 samples were collected from 500 patients. 111, 200, 84, and 103 patients were recruited from US, India, Sri Lanka, and Brazil, respectively. Mean age was 35 and 58% were men. The most common Ad subgenera were subgroup D (316, 63%), followed by E, B, then C. A significant portion of patients (110, 22%) lacked PCR evidence of Ad infection. There was a significant geographic bias for Ad subgenera and serotype distribution (Fisher exact, p-value <10-6). The odds ratio (OR) of having serotype 4 vs. others in the US was 7.87 times higher than in non-US sites (95%CI 3.55, 17.35), while the OR of having serotype 8 vs. others in the US was significantly lower (0.01, 95%CI 0.003, 0.043). Among the samples that underwent NGS, Ad was the dominant virus in 10 of 16 Ad-positive samples, while there was no consistent, dominant virus in 16 Ad-negative samples. Two Ad-negative samples contained significant amounts of torque teno virus. Conclusions: This study reveals unexpected geographic diversity in Ad genotypes associated with KC, and suggests that nearly one fourth of apparently viral KC may be due to agents other than adenovirus. . Commercial Relationships: Cecilia S. Lee, None; Aaron Y. Lee, None; Lakshmi Akileswaran, None; David W. Stroman, NovaBay Pharmaceuticals, Inc; Kathryn Najafi-Tagol, NovaBay Pharmaceuticals, Inc; Steve Kleiboeker; Anna Wald, None; Russell N. Van Gelder, None Support: NH Grant K23EY024921, R01EY022038, K24 AI071113, P30EY001730, Richard and Maude Charitable Foundation, Ferry Research Grant, Latham Vision Science Research Grant Clinical Trial: NCT01532336 These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Program Number: 3865 Poster Board Number: A0013 Presentation Time: 3:45 PM–5:30 PM Safety and efficacy study to evaluate wiping the lid margins with tap water alone or in combination with lid hygiene shampoo in subjects with normal meibomian glands Hirotaka Tanabe, Minako Kaido, Motoko Kawashima, Reiko Ishida, Kazuo Tsubota. Ophthalmology, Keio Univ School of Medicine, Tokyo, Japan. Purpose: We performed a prospective clinical study to evaluate the safety and efficacy of using tap water alone or in combination with lid hygiene shampoo to wipe lid margins with normal meibomian glands. Methods: Fourteen eyes of 7 subjects (6 males and 1 female aged 28 to 56 years old [40.8 ± 11.4 years old]) with normal meibomian glands were evaluated in the Ophthalmology Department of Keio University Hospital before and after wiping the lid margins using tap water alone at first and then tap water in combination with lid hygiene shampoo (Eye Shampoo [MediProduct, Japan]) one week later. The ethics committee of the Keio University School of Medicine approved the protocol. We used t-tests to analyze the tear break-up time (TBUT) (grades 0 to 10) and Wilcoxon signed-rank tests to analyze corneal and conjunctival fluorescein/lissamine green/ rose bengal staining scores (grades 0 to 9), lid margin lissamine green staining scores (grades 0 to 3), subjective symptoms (grades 0 to 100) (as assessed via the visual analog scale [VAS]), and tear lipid layer interference (grades 1 to 5) (as assessed using a DR-1 tear interference camera [Kowa Co., Nagoya, Japan]). Results: A significant exacerbating change (P<0.05) was not detected after either method was applied in terms of the VAS score, the TBUT, corneal and conjunctival staining, lid margin lissamine green staining, and tear lipid layer interference. For the VAS score, foreign body sensation and eye discharge recognition were significantly improved (P<0.05) only in the group using lid hygiene shampoo (9.71±14.0 to 3.92±9.02, P=0.031; 4.96±5.52 to 1.14±2.28, P=0.022, respectively), and eyestrain was significantly improved only in the group using tap water alone (17.0±15.3 to 1.57±4.01, P=0.008). Although there were no significant differences in the other VAS parameters, certain changes indicating improvements (P<0.1) in the parameters related to stimulation, i.e., pain and smarting pain, were noted only in the group using lid hygiene shampoo (3.64±6.18 to 0.64±1.64, P=0.098; 3.28±5.42 to 0.64±1.73, P=0.098, respectively). Conclusions: The use of tap water alone or in combination with lid hygiene shampoo to wipe lid margins appears to be safe for maintaining lid hygiene in normal subjects. The efficacy of lid hygiene shampoo seems greater than tap water alone based on the VAS parameters. Commercial Relationships: Hirotaka Tanabe, None; Minako Kaido, None; Motoko Kawashima; Reiko Ishida, None; Kazuo Tsubota, MediProduct (F) Support: MediProduct Clinical Trial: http://www.umin.ac.jp/ctr/index.htm, UMIN000016906 Program Number: 3866 Poster Board Number: A0014 Presentation Time: 3:45 PM–5:30 PM Histatin-1 as a peptide based therapy for human corneal epithelial wound healing Dhara Shah, Zeeshan Pasha, Marwan Ali, Vinay K. Aakalu. Department of Ophthalmology, University of Illinois at Chicago, Chicago, IL. Purpose: Purpose: Histatins are histidine-rich peptides that are present in human saliva and in human lacrimal epithelium. Previous experiments have shown that Histatin-1 can promote epithelialization in multiple tissue types. We investigated the effects of applying histatin-1 to a human corneal epithelial wound model in vitro. Methods: Methods: Human corneal epithelial (HCE) cells were utilized to perform a standard scratch based wound healing assay. A confluent monolayer of HCE cells were wounded manually by scraping with a 200ul pipette tip. Cells were then exposed to different concentrations of Histatin-1 in a medium essential media containing 2% serum. Cell migration was followed for 24 hours after scraping and imaged using time-lapse microscopy. Images were analyzed using Neuro Image-J. Effects of Histatin-1 on cell proliferation and cell death were assessed using colorimetric assays-lactate dehydrogenase (LDH) assay and an MTT assay. Results: Results: Our findings demonstrate that cell migration rates were increased with increasing Histatin-1 concentration. We also noted no significant change in LDH levels or MTT levels at tested levels, suggesting a non-proliferative, non-toxic mechanism for wound healing enhancement. Conclusions: Conclusion: Histatin-1 peptides promote human corneal epithelial migration and wound healing in vitro. Future studies to investigate these findings in vivo will be necessary to develop therapeutic applications for Histatin peptides. Human Corneal Epithelial cells 4 hour post scratch (Control) Human Corneal Epithelial cells 4 hour post scratch with Histatin-1 (10uM/ml) These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Commercial Relationships: Dhara Shah, None; Zeeshan Pasha, None; Marwan Ali, None; Vinay K. Aakalu, None Support: NIH—NEI Grant (K08EY024339), a seed grant from Illinois Society to Prevent Blindness, a Research Grant from Midwest Eye Banks, a Grant-in-Aid from Fight for Sight and Departmental Support through an NIH-NEI Core grant (2P30EY001792-36A1) and an Unrestricted Grant from Research to Prevent Blindness (NY,NY). Program Number: 3867 Poster Board Number: A0015 Presentation Time: 3:45 PM–5:30 PM Non-invasive technique for dynamic measurement of tear film surface quality based on interferometry Dorota H. Szczesna-Iskander, Slawomir Drobczynski. Dept. of Optics and Photonics, Wroclaw University of Technology, Wroclaw, Poland. Purpose: To demonstrate the development of a new generation Lateral Shearing Interferometer (LSI) for non-invasive assessment of tear film surface quality (TFSQ) dynamics. Methods: The LSI technique allows for objective in vivo evaluation of the tear film (TF) surface in the central area of a cornea or a contact lens by analyzing the shape of interference pattern. High sensitivity of this technique for subtle changes in TF dynamics has been demonstrated with a prototype (Szczesna et al. OVS 2009) whose several limitations hindered its potential to achieve full clinical utility. Recently, a new compact LSI was built. A HeNe laser (633nm) light source illuminates an area of 6 mm in diameter. The design of the interferometer meets the safety standards (ANSI Z136.1-2000). The LSI is fixed to a slit lamp base, the fixation target for the measured eye has been incorporated into the system, and a special head rest has been designed to minimize the head movements. Temporal dynamics of TF is recorded by a CCD camera with frequency of 29 frames per second. Results: Measurements on artificial eyes of radius from 7.8 to 8.5 mm (with and without astigmatism) as well as on over 50 subjects have been performed with the new LSI system as well as the previously used prototype. The slit-lamp based operation of the new LSI is more user friendly comparing to its predecessor. The new fixation target aligned with the measured eye rather than the fellow eye substantially improved eye’s stability during the measurement. The larger coverage area (6 mm vs 4 mm) and higher frequency of fringes determined by the optical wedge allow for more detailed analysis of the TFSQ and accurate assessment of TF post-blink stabilization phase and evolution of the TF break-up. The upward movement of the post-blink deterioration features is clearly observed (see figure). Conclusions: The new improved generation of LSI enhances its clinical utility for non-invasive assessment of TFSQ and its dynamic properties on cornea and contact lenses. The new LSI provides basis for studying relationships between TF post-blink stabilization phase and TF related eye diseases. Figure. Examples of interference patterns recorded with the new generation LSI: 0.10s (A); 0.34s (B); 0.68s (C); 1.00s (D) post-blink. The frames show the upward movement of the tear film features within the first second after the blink. Commercial Relationships: Dorota H. Szczesna-Iskander, None; Slawomir Drobczynski Support: National Science Center of Poland, Program SONATA, grand no: 2011/03/D/ST7/02512 Program Number: 3868 Poster Board Number: A0016 Presentation Time: 3:45 PM–5:30 PM Stimulation of Corneal Nociceptors Results in Pro-inflammatory Molecular and Cellular Responses Yashar Seyed-Razavi1, 2, Pedram Hamrah3, 4. 1Ophthalmology, Tufts Medical Center, Boston, MA; 2Ophthalmology, Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, MA; 3Ophthalmology, Schepens Eye Research Institute/Massachusetts Eye and Ear, Cornea & Refractive Surgery Service, Harvard Medical School, Boston, MA; 4Ophthalmology, Cornea Service, New England Eye Center, Tufts Medical Center, Boston, MA. Purpose: We have recently shown immediate changes in neuropeptides levels following CO2 stimulation of corneal nociceptors. The peripheral nervous system, in part through neuropeptides, is able to alter the balance towards a pro- or antiinflammatory environment by influencing the activity of resident antigen presenting cells (APCs) such as dendritic cells (DCs), modulating their chemotaxis, maturation and T-cell stimulatory capacity. We therefore hypothesized that CO2-induced nociceptor stimulation will result in alterations in downstream inflammatory cytokines and adhesion molecules, consistent with, and resulting in pain-induced neurogenic inflammation. Methods: CO2 gas was applied to the central cornea of C57BL/6 and BALB/c mice in a series of 3 pulses every hour over 4 hours, and collected 1 hour following the final burst. Corneas were then excised, and gene and protein analysis for pro-inflammatory cytokines and adhesion markers, as well as whole-mount staining for CD11b (leukocyte marker), CD11c (DCs) and MHC class II (antigen presenting cells), was performed. Naïve un-stimulated and airstimulated corneas served as controls. Results: Long-term stimulation of corneal nociceptors resulted in increased expression of the pro-inflammatory neuropeptides substance P (7-fold, p<0.001) and CGRP (3-fold, p<0.05), as well as the anti-inflammatory neuropeptide urocortin (70 fold, p<0.001) in C57BL/6 mice. IL-1β was up-regulated (3-fold, p<0.05), and whilst IL-6 expression increased, it was not significant (p=0.12) compared to controls. Increased expression of adhesion molecules P- and E-Selectin (p=0.059 and p<0.05, respectively) as well as elevated ICAM-1 expression (p<0.05) was found. Analysis of BALB/c mice revealed a similar increase in inflammatory cytokines (p=0.06 and p<0.01 for IL-1β and IL-6, respectively) and adhesion markers (p=0.053 and p<0.05 for E-Selectin and ICAM, respectively). Fractalkine (CX3CL1) mRNA levels increased (p<0.05) following CO2 stimulation. Finally, nociceptor stimulation resulted in alterations in resident APC density and maturation levels. Conclusions: Our study demonstrates downstream changes in inflammatory cytokines, chemokines and adhesion markers, ultimately leading to alterations in resident APC populations following nociceptor activation. To our knowledge, this is the first report to reveal direct neurogenic effect of pain-induced corneal nociceptor activation. Commercial Relationships: Yashar Seyed-Razavi; Pedram Hamrah, None Support: NIH R01-EY022695 These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Program Number: 3869 Poster Board Number: A0017 Presentation Time: 3:45 PM–5:30 PM Crosstalk Between Selective Autophagy and Nrf2-ARE Pathway Through p62/SQSTM1 Confers Limbal Stem Cell Resistance to Ultraviolet-A Irradiation Ying-Ting Chen1, Maria Laggner1, Florian Gruber2, Erwin Tschachler2, Ursula Schmidt-Erfurth1, Andreas Pollreisz1. 1 Department of Ophthalmology, Medical University of Vienna, Vienna, Austria; 2Department of Dermatology, Medical University of Vienna, Vienna, Austria. Purpose: Recently we reported a cytoprotective role for autophagy and Nrf2-ARE pathway in physiological UVA-stressed limbal stem cells (LSC). The current study sought to identify the mechanistic underpinnings linking these two pathways. Methods: Krt14-Cre:Atg7f/f transgenic mice and global Nrf2-KO mice on C57BL/6 background were used to generate Atg7-/- and Nrf2/LSCs, respectively. 3-MA was used to inhibit autophagy in cultured Nrf2-/- LSCs for creating autophagy-deficient Nrf2-KO LSCs. UVA irradiation at a low dose (10 J/cm2) was applied to the cultured LSCs, pre-treated with either ROS-scanvenger NAC or drug vehicle. CMH2DCFDA live staining and Caspase3/7 CytoEvent were used to detect intracellular ROS levels and cellular apoptosis, respectively. Autophagic activity was measured by CytoID live staining. Dual immunofluorescence was used to study the cellular kinetics of Nrf2 and p62/SQSTM1. A nuclear localization of Nrf2 was used as a surrogate readout for Nrf2-ARE activity. Results: Confocal microscopy using CM-H2DCFDA live staining did not detect changes of ROS level in 10 J/Cm2-irradiated Atg7f/f LSCs. In contrast, a 2-3 fold elevation in ROS level was observed in Atg7-/-, Nrf2-/- and autophagy-deficient Nrf2-/- LSCs (all p<.05). Compared to the perspective non-irradiated baseline, Caspase3(+) apoptotic cells was increased by 3.5% (p>.05), 21.7% (p<.05), 29.3% (p<.05), and 18.4% (p<.05) in Atg7f/f, Atg7-/-, Nrf2-/- and autophagydeficient Nrf2-/-LSC at 24-hr post-irradiation. The radiation-induced apoptosis was successfully rescued by NAC pre-treatment. Dual immunofluorescence revealed a diffuse cytosolic distribution for Nrf2 and p62 in unstressed LSCs, with a low-degree co-localization. After irradiation, Nrf2 nuclear translocation was observed exclusively in Atg7f/f cells with distinct p62 perinuclear speckle formation. Such a Nrf2/p62 redistribution was not seen in any other irradiated groups. Conclusions: Our data collectively suggest that autophagy activation in LSCs under physiological UVA stress functions as a non-canonical antioxidant defense mechanism. The p62/SQSTM1-mediated enhancement of Nrf2-ARE response enables LSC to have a tighter control in cellular redox. New therapeutics targeting p62/SQSTM1 might thus supplement the current antioxidant-based treatment for corneal degenerative diseases associated chronic oxidative stress. Commercial Relationships: Ying-Ting Chen, None; Maria Laggner; Florian Gruber, None; Erwin Tschachler, None; Ursula Schmidt-Erfurth, None; Andreas Pollreisz, None Program Number: 3870 Poster Board Number: A0018 Presentation Time: 3:45 PM–5:30 PM Ocular surface stories after cataract surgery Rodrigo M. Torres1, 2, Pablo G. Lódolo1, 2. 1Ophthalmology, Centro de Ojos Dr Lodolo, Colonia Avellaneda, Argentina; 2Asociación Entrerriana de Oftalmología, Paraná, Argentina. Purpose: “I can see but I’m felling bad”. When cataract surgeons heard something like that and they couldn’t find real problems in those eyes, usually the ocular surface (OS) service take place. This work review and analyze this topic. Methods: A retrospective clinical study was performed, evaluating the clinical charts from patients referred to the OS service with problems after cataract surgeries, during the present year (2015). The follow data was analyzed: age, sex, time after surgery, visual acuity, the kind of intraocular lens (IOL), post-surgical refraction, the patient complaints/symptoms, their work or occupation, previous topical treatments (dry eye/glaucoma), the discovered signs and their therapeutic approach. Results: A total of 15 patients operated (both eyes) by 5 different surgeons were evaluated. The mean age was 67 years all, with 9 women and 6 men. All of the cases were referred between 2 to 3 months after cataract surgery (sutureless clear corneal incision of 2.0 mm to 2.5 mm, topical anesthesia, without intraocular surgical or post-surgical complications). Binocular visual acuity without correction in all of the patients were between 20/25 to 20/20, but 12 of them with 1 eye for near sight (spherical refraction: -1.0 to -1.5). 3 patients with far sight/both eyes. Symptoms: pain (14), foreign body sensation (15), burning (12), itching (4); fear about blindness (15). Work/occupation: retired (10) professional work (5: architect/ physician/agronomic engineering/politic/bank employee). Non of the patients have history about glaucoma topical treatment but 3 have history about OS problems (dry eye:2; Stevens Johnson: 1). Signs: blepharitis (8), corneal edema around incision (4), epithelial keratitis (4), entropion (3). symblepharon (3). 5 patients from all (33.3%, 3 women, 2 men) don’t have signs about OS problems. Artificial tears, topical loteprednol and lid hygiene, were the therapeutic approach for most of them but five with oral minocycline 50mg and topical cyclosporin for three (two were pseudo-pemphigoid and one was the Stevens Johnson). Conclusions: Most women than men were referred to the OS service because complaints after cataract surgery. OS problems were detected in 66.6%, but fear about blindness was present in all the cases. These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Nonspecific pain was another common symptom. It is time to include a psychologist to evaluate pre-op patients and manage them at postop time, as part of the ophthalmology team? Commercial Relationships: Rodrigo M. Torres, None; Pablo G. Lódolo Program Number: 3871 Poster Board Number: A0019 Presentation Time: 3:45 PM–5:30 PM A Structural Equation Model (SEM) Relating Eye Pain (EP) and Ocular Surface Measurements (OSMs) Richard A. Bilonick1, 2, Anat Galor3. 1UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA; 2Biostatistics, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA; 3Bascom Palmer Eye Institute, University of Miami, Miami, FL. Purpose: Goals were to 1) parsimoniously describe the relationship between subjective EP (measured by questionnaire with 7 ordinal responses) and combination of objective OSMs and demographic data, and 2) use data from both eyes and describe the ordinal EP responses using a small number of latent factors. Methods: 190 patients underwent ocular surface assessment, including tear osmolarity (OSM), tear evaporation measured via tear breakup time (TBUT), corneal epithelial cell disruption measured via corneal staining (STN), and tear production measured via Schirmer’s strips with anesthesia (SCH). Subjects rated the intensity of their EP for right now (RN), worst 1 wk (W1W), average 1 wk (A1W), worst 3 months (W3M), average 3 months (A3M), worst 1 yr (W1Y), and average 1 yr (A1Y) using an ordinal rating scale anchored at 0 for no pain and 10 for most intense pain imaginable. 4 common factor (CF) pure measrement models (1-factor, 2-non-overlapping factors [2NF], 2 overlapping factors, and a 3-factor model) for EP responses were compared and the best measurement model for EP was chosen using root mean square error of approximation (RMSEA). OSM, TBUT, STN, SCH were measured in both eyes and were each represented by a latent factor with the left and right measurements as indicators. This approach accounted for random measurement error that would otherwise bias effect estimates. These latent factors along with age, sex and race were included as exogenous variables that could affect EP. R packages lavaan and OpenMx were used to estimate path coefficient and other SEM parameters. Results: 2NF CF model with latent factors STP (short term pain) and LTP (long term pain) was best according to RMSEA. The figure shows the path diagram for the resulting SEM. Correlations between each EP question response and corresponding STP and LTP all exceeded 0.9. RMSEA for the SEM was 0.04, 75% lower than for the 2NF CF pain model. A 100 mOsm/L increase in OSM resulted in a 1.5 standard deviation (SD) drop in LTP (p<0.001) and 0.5 SD drop in STP (p=0.016). While not SS, a 10 mm increase in SCH resulted in a 0.2 SD drop in LTP (p=0.144) and a 0.1 SD drop in STP (p=0.599). Conclusions: OSM was shown to affect both STP and LTP but in a differential manner. No evidence was found for effects due to TBUT, STN, age, sex or race. Possibly SCH had an effect which was also differential. Sem path diagram relating EP, OSMs, and demographics. Commercial Relationships: Richard A. Bilonick; Anat Galor, None Support: P30-EY008098; Eye and Ear Foundation (Pittsburgh, PA); Research to Prevent Blindness (New York, NY); Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development; Clinical Sciences Research and Development’s Career Development Award CDA-2-024-10S (Dr. Galor); NIH Center Core Grant P30EY014801. Program Number: 3872 Poster Board Number: A0020 Presentation Time: 3:45 PM–5:30 PM Human blinking ‘eye-on-a-chip’ Jeongyun Seo1, Woo Y. Byun1, Andrea Frank1, Mina Massaro -Giordano2, Vivian Lee2, Vatinee Y. Bunya2, Dongeun Huh1. 1Bioengineering, University of Pennsylvania, Phildelphia, PA; 2Scheie Eye Institute, Department of Ophthalmology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA. Purpose: The structural and environmental complexity of the ocular surface (OS) poses critical technical challenges to in vitro investigation of its physiology and pathology using traditional cell culture models. As a result, research in this area has relied heavily on expensive and time-consuming in vivo animal studies. To realize more physiological in vitro models, we have developed an ‘eye-ona-chip’ microdevice that recapitulates the three-dimensional (3D) tissue structure, dynamic mechanical environment, and physiological functionality of the human OS. Methods: Our microdevice (Fig. A) contains a 3D dome-shaped porous cell culture scaffold with a similar curvature to that of the human cornea (Fig. B). The porous scaffold was impregnated with human keratocytes (HKs) and collagen gel to mimic the stroma. We also developed a novel microengineering technique to precisely deposit human corneal epithelial cells (HCECs) and conjunctival epithelial cells (HCjEs) on the 3D scaffold surface to replicate the concentric epithelial patterns in vivo. These microengineered ocular tissues were integrated with a biomimetic hydrogel eyelid, and the eyelid was actuated to slide along the curved scaffold surface to replicate eye blinking. Results: The 3D tissue structures of the OS were reconstituted in our microdevice. The HKs were cultured within the porous scaffold to replicate the stroma (Fig. C). Our cell patterning technique allowed us to generate a circular corneal epithelium surrounded by a conjunctival epithelium on the curved scaffold surface (Fig. D). These ocular epithelia exhibited differentiated phenotypes as evidenced by stratification, tight junction formation, and increased cytokeratin/mucin protein expressions. We also demonstrated tear fluid dynamics in our microdevice by showing spreading of artificial These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts tear fluid and resultant wetting of the scaffold surface due to eyelid actuation (Fig. E). Finally, we explored the possibility of leveraging this microdevice to simulate pathological features of dry eye disease and to examine key biological processes mediating the development and progression of the disease. Conclusions: Our ‘eye-on-a-chip’ holds great potential to serve as an innovative modeling platform that represents a predictive and lowcost alternative to conventional cell culture and animal models. Commercial Relationships: Jeongyun Seo, None; Woo Y. Byun, None; Andrea Frank, None; Mina Massaro-Giordano, None; Vivian Lee, None; Vatinee Y. Bunya, None; Dongeun Huh Program Number: 3873 Poster Board Number: A0021 Presentation Time: 3:45 PM–5:30 PM A disease model of aqueous deficient dry eye in a microengineered ocular surface tear film platform Nicole Qiaozhi Lu, Michael P. Grant, Jennifer Elisseeff. Wilmer Eye Institute, Johns Hopkins University, Baltimroe, MD. Purpose: Dry eye is an ocular surface disease affecting a large population worldwide, and an in vitro model, recapitulating the function of ocular surface, is in need for drug development studies. Previously we have demonstrated the efficacy of cocultured lacrimal gland (LG) and conjunctival epithelial cells (CECs) in mimicking the tear secretory function of healthy ocular surface. Here we explored the possibility of utilizing this microengineered platform as a disease model of aqueous deficient dry eye. Methods: Rabbit LGs were digested, and spheroids were formed in a shaking flask. CECs were isolated by dispase II digestion. CECs were seeded on top of the Transwell insert and allowed to stratify under airlifting condition. LG spheroids were encapsulated in Matrigel and added to the periphery of the membrane (Fig. 1). Dry eye was introduced by the addition of either IL-1β or hyperosmolarity. Cyclosporin A (CsA) and dexamethasone (DEX) were used as treatments. The systems were maintained for 10 days in vitro. Samples were harvested and processed for the analyses of gene expression by qPCR, mucin secretion of CECs by texas-red conjugated dextran staining, LG secretion by β-hexosaminidase assay, and immunohistochemistry. Results: Exposure to IL-1β and hyperosmolarity increased the mRNA level of proinflammatory cytokines and chemokines (TNFα, IL-6 and MMP9), while treatments with CsA and DEX were able to reduce it. The model also reflected a change in epithelial morphology as shown in the H&E staining (Fig. 2a). The induced inflammation in the model significantly decreased epithelial stratification and led to more keratinization, and DEX was more effective in reversing this effect. Immunohistochemistry with mucin 5AC supported that mucin expression was also affected, and both CsA and DEX could restore the mucin level comparable to control (Fig. 2b). Similar trends were observed in LG and mucin secretions. Conclusions: This study has demonstrated that dry-eye like symptoms were observed in the in vitro model after the induction of inflammation, and administering of treatments were proven to be effective as well. We mimicked the physiological conditions of aqueous deficient dry eye in the cocultured system with CECs and LGs. Potentially this microengineered ocular surface platform could be further investigated for the screening of new dry eye therapeutics. Fig1 Fig2 Commercial Relationships: Nicole Qiaozhi Lu; Michael P. Grant, None; Jennifer Elisseeff, None Support: KKESH-Wilmer Research grant; Research to Prevent Blindness Program Number: 3874 Poster Board Number: A0022 Presentation Time: 3:45 PM–5:30 PM Contralateral investigation of postoperative refractive surgery inflammation: Small-incision Lenticule Extraction vs LASIK Marianthi Stergiou1, A. J. Kanellopoulos1, 2, George Asimellis1, 3. 1 LaserVision.gr Eye Institute, Athens, Greece; 2Ophthalmology, NYU Medical School, NY, NY; 3Kentucky College of Optometry, Pikeville, KY. Purpose: The purpose of this study was to comparative investigate potential impact of inflammation following myopic correction. The established current gold standard of femtosecond-assisted LASIK was compared to an all femto-second laser Small Incision refractive Lenticule Extraction (SMILE). No study has investigated the These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts potential differences between SMILE and LASIK from the standpoint of postoperative inflammation. Methods: This is a contralateral, perspective stud employing 10 consecutive myopic patients, in which one eye (group-A) was treated with the SMILE, while the other eye (group-B) with LASIK. The LASIK procedure employed the Alcon Refractive surgery platform (Alcon Surgical, Ft. Worth, TX) comprised of the FS200 femtosecond and the EX500 excimer laser. The SMILE procedure employed the 500 kHz VisuMax® femtosecond laser (Carl Zeiss Meditec AG, Jena, Germany). InflammaDry (Rapid Pathogen Screening, Inc., Sarasota, FL) is an in-office test that detects matrix metalloproteinase MMP-9, an inflammatory marker that is consistently elevated in the tears of patients with dry eye disease. Using direct sampling microfiltration technology, InflammaDry identifies elevated levels of MMP-9 protein in tear fluid samples taken from the palpebral conjunctiva. Levels of MMP-9 were tested 1-month, 3-months and 6-months postoperatively, facilitated with the use of InflammaDry solution. Inclusion criteria: 18 years of age or older. Exclusion Criteria: Allergy to cornstarch or Darcon, Allergy to topical anesthetic or fluorescein dye. Results: MMP-9 levels in group-A (LASIK) were 65±15 ng/ml, 55±12 ng/ml, 45±9 ng/ml at 1-, 3-, and 6- months, respectively. MMP-9 levels in group-B (SMILE) were 45±13 ng/ml, 42±12 ng/ ml, 37±8 ng/ml at 1-, 3-, and 6- months, respectively. All groups displayed statistically significant differences at the perspective follow-up periods examined. Conclusions: Potential differences between LASIK and SMILE include increased inflammation levels postoperatively up to 6 months. Commercial Relationships: Marianthi Stergiou, None; A. J. Kanellopoulos, i-Optics (C), ISP Surgical (C), Keramed (C), Avedro (C), Allergan (C); George Asimellis, None Program Number: 3875 Poster Board Number: A0023 Presentation Time: 3:45 PM–5:30 PM First Report of Simultaneous Double-Headed Pterygiectomy with Conjunctival Autograft, Amniotic Membrane and Fibrin Glue Jenny Ha1, Jesus A. Martinez1, 2, Yasaman Hosseini1, Michael Korchak2, Jayson Koppinger3, 2, Sandra L. Cremers1. 1 Visionary Ophthalmology, Rockville, MD; 2Ophthalmology, Medstar Georgetown University Hospital / Washington Hospital Center, Washington, DC; 3Georgetown University School of Medicine, Washington, MD. Purpose: There is a paucity of data regarding the safety of simultaneous double-headed pterygiectomy (SDP). We present a new suture and cautery free technique that uses both conjunctival autograft (CAG) and amniotic membrane graft (AM) over conjunctival defects. We evaluated the outcomes of these SDPs. Methods: This retrospective study examined the outcomes of 45 consecutive eyes of 43 patients who received SDP by one surgeon (JAM). Patients received 5 mg diazepam and intraoperative local anesthesia, in an office-based operating suite without an anesthesiologist. Pterygia bases were excised, extensive tenonectomy performed, and medial and lateral recti cleaned. The cornea was polished with a diamond burr and Mitomycin C 0.025% was applied for 2 min under the conjunctival edges using 6-8 soaked corneal shields. Over the temporal defect, a CAG measuring 1-2mm greater than the width (W) and anteroposterior (AP) of the defect was placed. A cryopreserved AM (AmnioGraft, Bio-Tissue) was used over the nasal defect. Both grafts were secured with fibrin glue (TISSEEL, Baxter Inc). Cauterization and sutures were avoided. Postoperative complications were recorded. Results: Of the 45 eyes, 2 were recurrent double headed pterygia at the time of surgery, previously performed by a different surgeon, and 1 eye was a recurrent nasal and primary temporal pterygia. The average nasal and temporal conjunctival defects were 7.4mm (limbal defect, L), 10.8mm (AP) and 11.2mm (W) and 7.3mm (L), 12.6mm (AP), and 12.0mm (W), respectively. One patient’s CAG was supplemented with AM over the remaining temporal defect due to insufficient superior bulbar conjunctiva; this patient did not develop recurrence. The mean follow up time was 39.6 weeks (range 4.6142.9w). There was 1 nasal recurrence (1.1%) from a primary case at 57w requiring a second CAG. One dellen (1.1%) and 4 granulomas (4.5%) were resolved with topical steroid drops and artificial tears. Conclusions: To our knowledge, this the largest study of simultaneous double-headed pterygiectomy. The use of conjunctival autograft over the temporal defect, cryopreserved amniotic membrane over the nasal defect, and fibrin glue in SDP eliminates trauma from sequential surgery. This procedure is safe and results in the one of the lowest recurrence rates demonstrated in the literature. a) Preoperative double headed pterygium. b) Postoperative outcome at 3 months. Commercial Relationships: Jenny Ha, None; Jesus A. Martinez, None; Yasaman Hosseini, None; Michael Korchak, None; Jayson Koppinger, None; Sandra L. Cremers, None Program Number: 3876 Poster Board Number: A0024 Presentation Time: 3:45 PM–5:30 PM The impact of meibomian gland dysfunction on the ocular surface parameters of glaucoma patients on long-term topical hypotensive medications Mehmet C. Mocan, Enes Uzunosmanoglu, Sibel Kocabeyoglu, Murat T. Irkec. Department of Ophthalmology, Hacettepe Univ School of Medicine, Ankara, Turkey. Purpose: Meibomian gland dysfunction (MGD) is a prevalent inflammatory eyelid condition that adversely affects tear stability and may potentially exacerbate ocular surface inflammation in patients on chronic topical glaucoma medications. The purpose of this crosssectional, observational study was to evaluate the impact of MGD on the ocular surface parameters of medically treated glaucoma patients. Methods: Seventy eyes of 70 glaucoma subjects with a mean age of 65.0±9.9 years (range=46-86 years) who were on long-term (>12 months) topical hypotensive medication were prospectively recruited. Patients with a history of ocular or periocular surgery, cicatrizing conjunctivitis and contact lens use were excluded. MGD was defined as the presence of signs consistent with meibomian gland terminal duct obstruction with or without accompanying resistance to meibomium expressibility. MGD was categorized between grades 1-5 according to clinical severity. Ocular surface disease index (OSDI) questionnaire was completed at the time of enrollment. Ocular surface tests consisting of tear break-up time (BUT), ocular surface staining with lissamine green (LG) and Schirmer test with anesthesia were employed. Student’s t test, chi-square test and Mann-Whitney U test were used in statistical comparisons. Forty-five healthy control subjects with no ocular disease were also included. These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Results: The mean duration of glaucoma was 12.0±9.0 years (range=1-35 years). Patients were on average 1.7±0.8 classes of glaucoma medication. MGD was detected in 56 (80.0%) glaucoma subjects. Of these 47 (83.9%) had signs consistent with mild to moderate MGD. The ocular surface test results of both glaucoma patients with MGD and those without MGD fared significantly worse (p<0.001) for all investigated parameters compared to those of healthy controls. However there were no significant differences between OSDI scores (p=0.912), BUT (p=0.635), LG staining scores (p=0.248), Schirmer test values (p=0.991), between glaucoma patients MGD versus those without MGD. Conclusions: Mild to moderate MGD is frequently encountered in medically treated glaucoma patients. However the presence of MGD does not appear to have an additional detrimental effect on the ocular surface to that already induced by chronic topical medication use. Commercial Relationships: Mehmet C. Mocan, None; Enes Uzunosmanoglu, None; Sibel Kocabeyoglu, None; Murat T. Irkec, None Program Number: 3877 Poster Board Number: A0025 Presentation Time: 3:45 PM–5:30 PM Microfluidic system based high throughput toxicity and efficacy screening system for eyedrops in ocular surface experiments Kyong Jin Cho2, Jongil Ju3, Rina Lee1, Dae Yu Kim5, 4, Jeongyun Kim1. 1Biomedical Science, Dankook University Graduate School, Cheonan, Korea (the Republic of); 2Ophthalmology, College of Medicine, Dankook University, Cheonan, Korea (the Republic of); 3Biomedical Engineering, College of Health Science, Korea University, Seoul, Korea (the Republic of); 4Biomedical Engineering, College of Medicine, Dankook University, Cheonan, Korea (the Republic of); 5Beckman Laser Institute Korea, Cheonan, Korea (the Republic of). Purpose: Toxicity and efficacy tests of eye drops is complex because the administration times and wash out duration of eyedrop should be considered. In addition, using a pipet tip on the corneal wound healing experiments, the size of the wound is not uniform and a physical injury is caused. We have developed a fully automated high throughput screening system based on the microfluidic cell culture array to evaluate toxicity and efficacy of eyedrops. Methods: The present micro fluidic chip used in the experiment is high throughput drug screening system and we can observe 8 step’ concentrations at the same time. These process are operated by fully automatic pneumatic controller that is engaged by the programmed LabVIEW software. We evaluated the toxic effect of benzalkonium chloride (BAK) and wound healing effect of ursolic acid on immortalized human corneal epithelial cell line (HCE-Ts) in this system. Results: 1. Micro fluidic high throughput screening (μHTDS) system for cytotoxicity of preservatives in eye drop : Cytotoxicity of BAK on HCE-Ts cells was increased depend on treatment time with comparing same dose conditions. Live cells with 10 minutes treatment are quickly decrease from 0.002% comparing to 5 minutes treatment results. The result in this study was similarity between plate experiment of conventional method and μHTDS system. 2. Micro fluidic high throughput screening system for wound healing (proliferation and invasion) model : 64 cell chambers which are arranged 8x8 forms, it means that each 8 concentrations can be observed on 8 chambers at same concentration. 7uM was most effective concentration of ursolic acid on migraion and invasion of HCE-Ts cells. Conclusions: We have developed a micro fluidic high throughput drug screening (μHTDS) system for cytotoxicity and efficacy screening of eyedrops with 8 different drug concentrations. Our μHTDS system can obtain more progressed cytotoxicity and efficacy results with minimizing the consumption of reagent, time, and labor. This system can be useful for cytotoxicity and efficacy screening the new drug against diverse cell type instead of human eye. Commercial Relationships: Kyong Jin Cho; Jongil Ju, None; Rina Lee, None; Dae Yu Kim, None; Jeongyun Kim, None Support: This work was supported by grants (NRF2013R1A1A1010734) from Ministry of Education. Program Number: 3878 Poster Board Number: A0026 Presentation Time: 3:45 PM–5:30 PM Signal detection theory examination of ocular surface sensory processing of corneal pneumatic stimuli Varadharajan Jayakumar, Trefford L. Simpson. University of Waterloo, Waterloo, ON, Canada. Purpose: To evaluate the feasibility of using signal detection theory (SDT) to examine the ocular surface sensory processing of corneal pneumatic mechanical stimuli using the Waterloo Belmonte aesthesiometer. Methods: Ten asymptomatic participants were recruited in this study. All participants were naïve to the SDT part of the experiment, but 6 participants were familiar with the other experiments using esthesiometer. The pneumatic stimulus was at eye temperature (approx. 33°C) and to change strength, flow was systematically varied. The stimulus intensities for the SDT experiment were obtained using the ascending method of limits, in which three ascending runs were averaged to estimate the threshold. Then, to keep the signal constant for each participant, flow was set at 1.5X threshold. 100 trials (demarcated using auditory cues) were presented with a signal probability of 0.4 (i.e., 60 catch trials with stimulus intensity of zero) and with signals and catch trials in random order. Responses (“yes” (there was a signal) or “no” (there was no signal)) were recorded using a button box with auditory feedback for each response. Breaks occurred after approx. 50 trials. Participants were instructed to blink normally between stimuli, and training was provided before the data were used. Detectability (d’), criterion (c) and likelihood ratio (β) were calculated using false alarms and hit rates. Results: The average (± SE) d’ was 0.52 (± 0.12), criterion was 0.22 (± 0.10) and β was 1.11 (± 0.06). The area under the curve obtained was 0.64 (±0.03). In addition, non-parametric SDT metrics were calculated. The average A’ was 0.67 (±0.03) and the non-parametric response bias β” was 0.07 (±0.03). Conclusions: The experiment shows the feasibility of using a SDT approach to examine ocular surface sensory processing with less concern about the potentially important impact of participant criterion These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts changing threshold and masking meaningful effects. The data suggest that participants in this experiment chose a relatively conservative strategy (reporting ‘no’ to trials more commonly) but this might be anticipated considering we designed the experiment with a relatively large proportion of catch trials. The data also suggested that some of the assumptions about classical SDT (underlying homoscedastic Gaussian ‘signal + noise’ and ‘noise’ distributions) need to be examined. Commercial Relationships: Varadharajan Jayakumar, None; Trefford L. Simpson, None Support: NSERC Canada, CFI Program Number: 3879 Poster Board Number: A0027 Presentation Time: 3:45 PM–5:30 PM Ocular Involvement In Sjogren’s syndrome – The Singapore Sjogren’s Syndrome Study Rachel Lim, Dinesh Gunasekeran, Jeggrey Kam, Bernard Thong, Rupesh V. Agrawal. Tan Tock Seng Hospital, Singapore, Singapore. Purpose: Sjogren’s syndrome may present with both ocular and extraocular manifestations. This study aims to compare and contrast ocular and extraocular manifestations in consecutive patients with Sjogren’s Syndrome (SS) recruited from rheumatology and ophthalmology outpatient clinics in Singapore. Methods: Computerized Physician Order Entry records of patients with physician-diagnosed SS between 1993 and 2013 were retrospectively analyzed. Patients were grouped into different categories from 1 to 10 based on the number of organ-systems involved including ocular involvement. Results: A total of 355 patients were included, with a majority being females(94.6%); and Chinese (84.5%). Mean duration of disease was 8±4 years. While 135 (38.0%) fulfilled American European Consensus Group (AECG) criteria for diagnosis, only 24 (6.8%) fulfilled Sjögren’s International Collaborative Clinical Alliance (SICCA) criteria for SS. Primary-SS comprised 76.9%, secondarySS 23.1%; with the most prevalent associations of rheumatoid arthritis (11.8%) and systemic lupus erythematosus (11.3%). The most common ocular manifestations were keratoconjunctivitis sicca (KCS) (91.0%), anterior uveitis (2.54%), episcleritis (2.54%), lacrimal gland enlargement (1.69%) and retinal vasculitis (0.28%). The most common extraocular manifestations were musculoskeletal (75.2%), lymphoreticular (47.6%) and constitutional (41.7%). All patients had at least 1 other extraocular organ-system involved, with median number of systems affected being 4 in 26% patients, and maximum of 10 organ-systems affected in 1 (0.28%) patient. Ocular-musculoskeletal involvement was the most common combination (244 patients, 68.73%). Anti nuclear antibody (ANA) was tested positive in 298/343 (86.9%) with titres of ≥ 1:320 in 193/298 (64.8%), rheumatoid factor (RF) was positive in 191/280 (68.2%), anti-Ro in 241/350 (68.9%), and anti-La in 90/314 (41%). Hydroxychloroquine (83.4%), methotrexate (14.4%) and azathioprine (12.4%) were the most commonly used therapies. Conclusions: Keratoconjunctivitis sicca is the commonest ocular manifestation of Sjogren’s syndrome. 90% of the patients had two or more organ-systems involved, with musculoskeletal involvement being the most common. Commercial Relationships: Rachel Lim, None; Dinesh Gunasekeran, None; Jeggrey Kam, None; Bernard Thong, None; Rupesh V. Agrawal, None Program Number: 3880 Poster Board Number: A0028 Presentation Time: 3:45 PM–5:30 PM HLA class I genes associated with Cold Medicine related StevensJohnson Syndrome with Severe Ocular Complications in the Brazilian population Tais H. Wakamatsu1, Mayumi Ueta2, Katsushi Tokunaga3, Yukinori Okada4, Renata R. Loureiro1, Karita A. Costa1, Juliana M. Sallum1, José Milhomens1, Chie Sotozono5, Jose Alvaro P. Gomes1, Shigeru Kinoshita5. 1Federal University of Sao Paulo, Sao Paulo, Brazil; 2Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan; 3Department of Human Genetics, University of Tokyo, Tokyo, Japan; 4Human Genetics and Disease Diversity, Tokyo Medical and Dental University, Tokyo, Japan; 5Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan. Purpose: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are immune-complex mediated hypersensitivity diseases that typically involve the skin and mucous membranes. Here we investigated etiologic factors such as causative drugs and the association between human leukocyte antigen (HLA) class I genes and cold medicine-related SJS/TEN (CM-SJS/TEN) with severe ocular complications (SOC) in Brazilian patients. Methods: We enrolled 74 Brazilian patients with SJS/TEN with SOC and 135 healthy Brazilian volunteers. The patients’ ethnicity was Pardo (51%), European ancestry (40%) and others. All were interviewed with respect to possible etiologic factors. HLA class I (HLA-A, -B, -C) were examined to determine whether there was a genetic predisposition for CM-SJS/TEN with SOC. Results: We identified cold medicine as the main causative drug (39 patients - 53%). HLA-A*66:01, HLA-B*44:03 and HLA-C*12:03 were associated, and HLA-A*11:01, HLA-B*08:01, and HLA-B*51:01 were inversely associated with Brazilian CMSJS/TEN with SOC. After dividing in Pardo and European ancestry, HLA-A*66:01 was associated with among individuals both Pardo and European ancestry; HLA-B*44:03 and HLA-C*12:03 among individuals with only European ancestry. Conclusions: In conclusion, our findings suggested that HLA-A*66:01 might be a marker in Pardo and European ancestry and HLA-B*44:03 and HLA-C*12:03 might be markers in only European ancestry. Moreover, HLA-A*11:01 might be a marker of resistance to CM-SJS/TEN with SOC. Commercial Relationships: Tais H. Wakamatsu, None; Mayumi Ueta, None; Katsushi Tokunaga, None; Yukinori Okada, None; Renata R. Loureiro, None; Karita A. Costa, None; Juliana M. Sallum, None; José Milhomens, None; Chie Sotozono, None; Jose Alvaro P. Gomes, None; Shigeru Kinoshita, None Program Number: 3881 Poster Board Number: A0029 Presentation Time: 3:45 PM–5:30 PM EVALUATION OF CHANGES IN OCULAR SURFACE AFTER VITRECTOMY SURGERY Chiara Del Noce, Francesca Bruzzone, Silvio Lai, Maurizio . ROLANDO, Carlo E. Traverso. DINOGMI, University of Genoa Eye Clinic, Genoa, Italy. Purpose: To evaluate the relationship between vitrectomy surgery and the alteration of ocular surface homeostasis. Methods: A total of 25 phakic patients with vitreo-retinal interface pathologies and without ocular surface diseases were enrolled. All underwent the conventional 25/23 gauge pars plana vitrectomy surgery. Evaluation were made using a VAS symptoms questionnaire (subjective parameters scored from 0 to 10) and a slit lamp (blink These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts rate, palpebral and bulbar conjunctival irritation, BUT, meniscus, fluorescine and lissamine green staining). Results: One year after surgery, according to the VAS classification the ranking resulted: desire to keep the eyes closed, foreign body sensation, burning, pain, roughness and heat. Blink was incomplete and more frequent in 32 % of patients. Approximately 98% of patients revealed both palpebral and conjunctival irritation. BUT was reduced in 80% of patients. The meniscus was irregular and thinned in 88% of patients. According to the Oxford scale, fluorescein staining was positive in 60% of patients: 73% Grade I and 27% Grade II while lissamine green staining was positive in 84% of patients: 52% of grade I, 28% grade II and 20% of grade III. Conclusions: Vitrectomy surgery 23/25 gauge with transconjunctival approach might give ocular surface alterations and ocular discomfort. Commercial Relationships: Chiara Del Noce, None; Francesca Bruzzone, None; Silvio Lai, None; Maurizio ROLANDO, None; Carlo E. Traverso, None Program Number: 3882 Poster Board Number: A0030 Presentation Time: 3:45 PM–5:30 PM Deletion of the Vitamin D Receptor Affects Meibomian Gland of Mice Kai Jin1, Motoko Kawashima1, Masataka Ito2, Kokoro Sano1, Kazuo Tsubota1. 1Ophthalmology, Keio University, school of medicine, Tokyo, Japan; 2National Defense Medical College, Saitama, Japan. Purpose: The vitamin D receptor (VDR) and vitamin D metabolic associated enzymes have been determined in various kinds of cells in both mouse eyes and human eyes. In this study, we examined the effects of VDR knockout on the pathological changes of meibomian gland (MG), eyelid tissue, and the secretion of tear fluid. Methods: Eyelids of VDR knockout mice aged 3 months (n=10) and 7 months (n=10) were investigated and compared with 20 normal eyelids from age-matched wild type C57/B6J mice. Microscopy examination and fluorescein staining were performed to check the ocular surface. Cotton thread test, MILLIPLEX® MAP and qRTPCR were used to analyze the tear fluid and eyelid tissue. We also examined the haemotoxylin and eosin staining of the eyelid and lacrimal gland (LG), and observed the immunofluorescent staining for cytokeratins CK1, CK5, CK6. Results: Keratinous cysts of MG, partial dropout of MG, and thinning and whitening of the cilia were shown in VDR knockout mice aged 3 months or 7 months. With aging, MG cysts progressively grew while the obstruction, lump or nodule of MG on the conjunctival surface can be observed. The secretion of tear fluid from VDR knockout mice significantly increased (P<0.05) and mRNA expression of mucin and some inflammatory markers in eyelid tissue were also upregulated significantly (P<0.05). The lid margin of VDR knockout mice showed more expression of CK1. In addition, there were no significant differences of corneal fluorescein staining or haemotoxylin and eosin staining of the lacrimal gland (LG). Conclusions: Vitamin D receptor knockout influences the pathogenesis of MG, hyper-keratinization of the eyelid, and secretion of tear fluid. Commercial Relationships: Kai Jin; Motoko Kawashima, None; Masataka Ito, None; Kokoro Sano, None; Kazuo Tsubota, None Program Number: 3883 Poster Board Number: A0031 Presentation Time: 3:45 PM–5:30 PM Calculating the health economic burden of microbial keratitis (MK) admission in a tertiary referral centre in the UK Jasvir Virdee1, George Moussa1, Nicholas Gooch2, Jesse Kigozi2, Cristina Penaloza2, Saaeha Rauz1. 1Academic Unit of Ophthalmology, The University of Birmingham, Birmingham, United Kingdom; 2Health Economics Unit, The University of Birmingham, Birmingham, United Kingdom. Purpose: The UK health care system (National Health Service, NHS) provides medical care that is available to all and free for all. MK is the commonest ophthalmic emergency admission in the developed world, with large cost burden to the NHS and few health economic studies. This study was designed to provide a quantification of direct costs of inpatient care for MK versus income generated though coding in a supra-regional tertiary centre in the UK. Methods: Extensive clinical, demographic and economic data were collected retrospectively for a period of twelve months (Jan-Dec 2013) for 101 consecutive patients admitted with MK on a validated electronic proforma. Direct cost of admission (COA) was calculated using national reference costs for individual patients for various parameters including length of stay in days (LOS), topical medication, cost of microbiological services and cost of an ophthalmic hospital bed, together with health economic analytical assumptions to generate profit/deficit profiles based upon actual income and estimated expenditure A one-way ANOVA analysis was performed to compare groups. Results: The total income generated through discharge coding for all patients was £267,028, whilst calculated cost of admission was £382,473, giving an overall deficit of £115,445 per annum. The median individual deficit was £779 (interquartlie range £1880). The most critical factor driving the cost deficit was length of stay with median cost neutrality achieved between days 5 and 6 (table 1). Severity of microbial keratitis (graded according to Keay et al) was not found to be a significant factor in driving costs (Table 2). Surgical intervention (corneal gluing, evisceration) drove up costs of care. Conclusions: Health economic analysis shows that length of stay is the key driver for direct costs of care for patients admitted with microbial keratitis with the pivotal LOS of 4 days. The microbial keratitis treatment pathway should encourage discharge of patients who are able to selfadminister treatment after the sterilisation phase to enable financial parity. Prospective data collection is required to refine direct cost analyses and to evaluate the clinical and financial burden (indirect costs) of the impact of visual morbidity on quality of life. These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Commercial Relationships: Jasvir Virdee, None; George Moussa, None; Nicholas Gooch, None; Jesse Kigozi; Cristina Penaloza, None; Saaeha Rauz, None Program Number: 3884 Poster Board Number: A0032 Presentation Time: 3:45 PM–5:30 PM Using corneal chemical detection thresholds to select subjects for ocular surface discrimination sensory panels Trefford L. Simpson4, 1, Nancy J. Keir2, William Ngo3, 1, Yunwei Feng4, 1. 1University of Waterloo, Waterloo, ON, Canada; 2 Cooper Vision, Pleasanton, CA; 3School of Optometry & Vision Science, Centre for Contact Lens Research, Waterloo, ON, Canada; 4 School of Optometry & Vision Science, Waterloo, ON, Canada. Purpose: To determine if we can we select sensitive (‘good’) and less sensitive (‘bad’) participants in a sensory panel examining interocular judgments of contact lens comfort, using Belmonte esthesiometry. Methods: 49 subjects (adapted contact lens wearers) had chemical detection thresholds (% added CO2) measured with the Waterloo Belmonte Esthesiometer, on 3 occasions (at least a day apart), using the ascending method of limits. From the averages of these 3 measurements, 25 subjects were grouped as more sensitive (thresholds at or below median) or less sensitive (thresholds above median). Subjects also interocularly matched the discomfort of 3 optimally fit contact lenses using the mechanical or chemical stimuli of the esthesiometer and scaled discomfort using magnitude estimation. We tested a simple hypothesis: Subjects in the sensitive group would be better at interocular matching - regression pneumatic stimulus match to rated discomfort would be linear with a positive slope, but the control group would not perform well, with random slopes (positive, 0 and negative). Bayesian mixed modeling (with vague priors) estimated the group slopes (among others) and the distributions of these estimates were compared in the sensitive and control groups defined by chemical detection thresholds. R and rjags were used for the data analysis. Results: For the ‘bad’ group the regression slopes mode was 0.05 and the 95% HDI was -0.526-0.731. The figure (of the posterior distribution of the slope estimates) shows that for the ‘good’ group the mode was shifted and the 95% HDI did NOT include zero. Conclusions: Chemical thresholds can be used to separate sensitive and insensitive subjects who can match contact lens discomfort in a ‘well behaved’ way and therefore pneumatic esthesiometry may be used to select sensory panels. Commercial Relationships: Trefford L. Simpson, Alcon (F); Nancy J. Keir, None; William Ngo, None; Yunwei Feng Support: NSERC Canada, CFI, Alcon IIS. Program Number: 3885 Poster Board Number: A0033 Presentation Time: 3:45 PM–5:30 PM Ocular Surface Homeostatic Surveillance by Th17 Cells in the Closed Eye Cameron K. Postnikoff, Kelly K. Nichols. Vision Sciences, University of Alabama at Birmingham, Birmingham, AL. Purpose: T cell infiltration of the ocular surface is commonly only associated with inflammation and diseases such as dry eye. Neutrophils are well known to populate the closed eye during eyelid closure, and have been shown to signal the adaptive immune response in other organ systems. The purpose of this investigation was to catalogue and phenotype various leukocytes of the closed eye. Methods: Eleven healthy participants were recruited and were trained to wash the ocular surface with phosphate buffered saline for at-home self-collection of tear film leukocytes following a full night of sleep. Cells were isolated and counted and were incubated with fluorescently-labeled antibodies against CD45, CD14, CD15, CD16, CD11b to identify neutrophils and monocytes. Antibodies against CD45, CD3, CD4, CD8, CD196, and CD161 were used to identify T cells. A Becton Dickinson (BD; San Jose, CA) proprietary fixable viability stain was used to exclude dead cells from analysis. A BD LSR II flow cytometer was used for all analysis. Results: Neutrophils were the most dominant of all cell populations, representing roughly 58% of all leukocytes. Monocytes were not readily observed, representing only a small portion of cells (<2%). Both CD4+ and CD8+ T cells were found in the ocular surface eyewash after a full night of sleep. CD4+ were more abundant, representing approximately 7% of the total leukocyte population, while CD8+ cells were roughly 1.5% of the total leukocyte population. Finally, Th17 cells, as identified as being CD196+CD161+ represented, on average, 20% of the total CD4+ T cells. Conclusions: This investigation demonstrated T cell surveillance of the ocular surface, during sleep. Combined neutrophil and T These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts cell infiltration suggests that closed eye homeostasis is complex, and future studies seek to identify the mechanisms of leukocyte interaction and activation, and ultimately determine how dysregulation may contribute to disease. Commercial Relationships: Cameron K. Postnikoff, None; Kelly K. Nichols, None Support: Thank you to Dr. Karen Ersland for her assistance with flow cytometry. The authors also acknowledge the UAB Comprehensive Flow Cytometry Core, which is supported by NIH P30 AR048311 and NIH P30 AI27667. Program Number: 3886 Poster Board Number: A0034 Presentation Time: 3:45 PM–5:30 PM GRADING BULBAR REDNESS USING THE KERATOGRAPH 5M. CORRELATIONS WITH EFRON AND MCMONNIES SCALES Néstor Ventura Abreu1, Francisco Pérez Bartolomé1, Claudia Sanz Pozo1, Jose María Martínez de la Casa1, Javier Moreno-Montanes2. 1 Ophthalmology, Hospital Clínico San Carlos, Madrid, Spain; 2 Ophthalmology, Clínica Universitaria de Navarra, Pamplona, Spain. Purpose: To examine interobserver reproducibility of the Efron and McMonnies bulbar redness (BR) scales and establish correlations between these scales and the new Keratograph 5M. Methods: This was an observational cross- sectional study. 203 eyes of 203 subjects (50 controls, 153 under treatment with topical hypotensive drugs for glaucoma) with varying degrees of conjunctival hyperaemia were scored automatically for BR using the Keratograph 5M. These scores were then correlated with the gradings provided by two image-based subjective scales: Efron and McMonnies. The interobserver reproducibility of both scales was also evaluated. Results: Excellent reproducibility was observed for both the Efron (Weighted K= 0.897, 95% CI 0.823 – 0.904) and McMonnies (Weighted K= 0.783, 95% CI 0.752 – 0.795) scales. Keratograph BR scores (overall redness) and the scores obtained with both Efron (Spearman’s Rho= 0.43, P<0.001) and McMonnies (Spearman’s Rho= 0.48, P< 0.001) were significantly correlated. Redness scores provided for the bulbar and limbar nasal and temporal quadrants also correlated well with the two subjective scales. Conclusions: The Keratograph 5M method of assessing BR seems to be a good alternative to the subjective interpretation of image-based BR scales. Commercial Relationships: Néstor Ventura Abreu, None; Francisco Pérez Bartolomé, None; Claudia Sanz Pozo, None; Jose María Martínez de la Casa, None; Javier Moreno-Montanes, None Program Number: 3887 Poster Board Number: A0035 Presentation Time: 3:45 PM–5:30 PM Novel pterygium animal model using White New Zealand rabbit and murine fibroblasts (NIH 3T3 cell line) Julio C. Hernandez2, Jorge E. Valdez2, Judith Zavala2, Jorge Valenzuela1. 1Tecnologico de Monterrey, School of Medicine, Monterrey, Mexico; 2Ophthalmology and Visual Sciences Research Chair, Tecnologico de Monterrey School of Medicine, MONTERREY, Mexico. Purpose: To develop an animal model for pterygium using subconjunctival injection of murine fibroblast cells (NIH 3T3 cell line) in white New Zealand rabbits Methods: Under general and topical anesthesia with tetracaine hydrochloride, six 3 month-old white New Zealand rabbits received subconjuntival injection of 20,000 murine fibroblast cells (NIH 3T3 cell line) and 5 µL of Matrigel(Corning Inc, NY, US) on the perilimbal temporal bulbar conjunctiva of the right eye using a 1ml 30G needle. Left eyes were injected with 5 µL of Matrigel under the perilimbal temporal bulbar conjunctiva (control). Eyes were photographed under a magnification of 16x using a 12 megapixel digital camera attached to the microscope (WPI, Inc. FL, US) on day 1, 3 and 7. Conjunctival vascularization was measured on day 1,3 and 7 by analyzing images using Adobe PhotoshopCS5 (Adobe Inc, San Jose, CA) color histograms to measure red pixel saturation on a 6x6 mm area on the site of injection. Area of corneal and conjunctival fibrovascular tissue formation was assessed by analyzing the images on day 3 and 7 using area measurement software (Adobe Photoshop CS5). Statistical analysis was performed using t-test for mean comparison, statistical significance was considered with a p value <.05 Results: Red pixel saturation for right and left eyes was 102.53±11.07pxs and 104.84±8.34pxs (p=.609), 176.15±6.35px and 168.29±18.89pxs (p=.325), 221.58±33.85pxs and 168.70±17.75pxs (p=.006) on day 1, 3 and 7 respectively. Area of fibrovascular tissue on the site of injection in the right and left eyes was 14.32±3.47mm2 and 8.41±2.08mm2 (p=.002), 41.18±4.40 mm2 and 14.69±2.81 mm2 (p<.001) on day 3 and 7 respectively. Significant difference was observed between day 3 and 7 on right eyes (p<.001) and left eyes (p=.002) when comparing the area of fibrovascular growth. Red pixel saturation was higher for both eyes on day 7 when compared to day 3, but was only the different was only significant for right eyes (p=.017) Conclusions: Subconjunctival injection of murine fibroblast cells and Matrigel in white New Zealand rabbits was more effective for inducing conjunctival vascularization and fibrovascular tissue growth on the site of injection and surrounding cornea than subconjunctival Matrigel alone. A novel animal model for pterygium is presented with potential use for research on molecular mechanisms of pathogenesis and new treatment strategies. Commercial Relationships: Julio C. Hernandez, None; Jorge E. Valdez, None; Judith Zavala, None; Jorge Valenzuela, None Program Number: 3888 Poster Board Number: A0036 Presentation Time: 3:45 PM–5:30 PM Lax Eyelid Syndrome, Obstructive Sleep Apnea Syndrome (OSAS), and Ocular Surface Inflammation Mackenzie Becker1, Clayton Kirk1, Ramsudha Narala2, Sunita Kumar1, William Adams1, Charles S. Bouchard1. 1 Ophthalmology, Loyola University Medical Center, Maywood, IL; 2 Kresge, Detroit, MI. Purpose: Lax eyelid syndrome (LES) describes the association of distensible “floppy” eyelids and chronic papillary conjunctivitis that These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts has a well-known association with OSAS. Eyelid histopathology suggests that LES may result from overexpression of matrix metalloproteinase 9 (MMP-9) related elastin degradation. However, the relationship between the degree of eyelid laxity and OSAS and elucidation of MMP-9 levels in LES subjects has not been clearly demonstrated. The purpose of this study therefore was: 1) establish the prevalence and degree of eyelid laxity in patients with newly diagnosed OSA; 2) to evaluate the presence of MMP-9 in the tear film of patients with LES; and 3) to compare the current LES grading systems with a newly designed “laxometer.” Methods: Following OSAS evaluation by polysomnography, subjects were referred for eyelid laxity determination and an MMP tear assay. Measurements of the degree of eyelid laxity were determined for each eye using the following four methods: 1) degree of tarsal conjunctiva exposure; 2) ease of upper eyelid eversion; 3) degree of medial canthal tendon laxity; and 4) distensability of the laxometer. The MMP-9 tear film assay was conducted using a commercially available assay. Results: There was a small but non-significant positive association between laxometer measurement and duration of eyelid eversion (τ = 0.16, p = .29). Conversely, there was a small but non-significant negative association between laxometer measurement and duration of eyelid eversion (τ = -0.19, p = .25). A nonparametric Kreskas-Wallis test was used to assess whether eyelid elasticity varied as a function of sleep apnea severity. Although the study was unpowered for statistical significance, a positive trend was found between degree of sleep apnea and eyelid laxity determined by laxometer measurements. For the MMP-9 levels, 14 of 16 eyes (89%) with LES had a positive MMP-9 assay (p< .001). Conclusions: Although our current study sample is small, the laxometer data suggests an association between the severity of LES and the severity of OSAS that may have clinical relevance. The findings of elevated MMP-9 supports a notion that MMP plays a role in elastin degradation that may be associated with chronic conjunctivitis and reactive ocular surface typically found in patients with LES. We are currently recruiting additional patients to better assess the relationship between lid laxity and OSAS. Commercial Relationships: Mackenzie Becker, None; Clayton Kirk, None; Ramsudha Narala, None; Sunita Kumar, None; William Adams, None; Charles S. Bouchard, None Support: Richard A. Perritt Charitable Foundation Program Number: 3889 Poster Board Number: A0037 Presentation Time: 3:45 PM–5:30 PM A novel murine behavioral model of ocular neuropathic pain Romulo Albuquerque1, Jooyoung Cho1, Bradley Taylor2, Jayakrishna Ambati1. 1Ophthalmology, University of Kentucky, Lexington, KY; 2Physiology, University of Kentucky, Lexington, KY. Purpose: Neuropathic pain is a complex chronic state arising from injured and dysfunctional nerve tissue that is poorly responsive to analgesic drugs. Neuropathic pain also affects the eye. Ocular neuropathic pain, disguised as ocular surface disease, is often unrecognized and undertreated by clinicians (Rosenthal and Borsook 2015). We tested the hypothesis that alkali burn injury to the ocular surface leads to neuropathic changes in the ocular-trigeminal system and can be used as a molecular and behavioral model to study ocular neuropathic pain. Methods: Wild-type C57Bl6 mice were used. The alkali burn injury model was performed as previously shown (Anderson et al. 2014). Satellite glia activation and neuronal sensitization were studied in the trigeminal ganglia by immunohistochemistry using glial fibrillary acidic protein (GFAP) and neuropeptide-Y (NP-Y). Ocular pain behavior was assessed by eye wiping behavior after application of hypertonic (5M) saline solution to the ocular surface. Ocular pain behavior was tested before and after intra-peritoneal gabapentin (100mg/kg) administration. Results: Alkali injury to the ocular surface induced expression of GFAP and NP-Y in trigeminal ganglion satellite glial cells and neurons respectively. Alkali injury also increased eye wiping behavior in the injured eye, but not in the contralateral eye up to one week after injury. Increased eye wiping behavior was significantly reduced by treatment with intra-peritoneal gabapentin. Conclusions: Our model combines two previous ophthalmology models: a surface alkali burn injury to model ocular inflammation (Anderson, Zhou et al. 2014), together with an ocular pain testing involving topical application of hypertonic saline to elicit an eyewiping response, a behavioral index of pain. Here we show that eyewiping behavior on the eye ipsilateral but not contralateral to alkali burn was augmented, suggesting sensitization of ocular nociception. These behavior changes were paralleled by increased levels of GFAP and NP-Y in the trigeminal ganglia satellite glial cells and neurons. Eye wiping behavior was abrogated by systemic administration of gabapentin, an antiseizure drug used for the treatment of neuropathic pain. The ability of an analgesic drug like gabapentin to decrease augmented eye wiping suggests that this response is neuropathic in nature and provides a novel model to study the molecular mechanisms of ocular neuropathic pain. Commercial Relationships: Romulo Albuquerque; Jooyoung Cho, None; Bradley Taylor, None; Jayakrishna Ambati, iVeena Pharmaceuticals (I), Inflammasome Therapeutics (I), iVeena Holdings (I), iVeena Pharmaceuticals (S), Inflammasome Therapeutics (S), University of Kentucky (P), iVeena Holdings (P), iVeena Pharmaceuticals (P), iVeena Holdings (S), Allergan (R), Olix Pharmaceuticals (F), iVeena Delivery Systems (I), iVeena Delivery Systems (S), iVeena Delivery Systems (P) Program Number: 3890 Poster Board Number: A0038 Presentation Time: 3:45 PM–5:30 PM Prosthetic Replacement of the Ocular Surface Ecosystem Therapy for Sjogren’s Syndrome Patients Billy X. Pan, Gloria B. Chiu, Martin Heur. Ophthalmology, USC Eye Institute, Los Angeles, CA. Purpose: Sjogren’s syndrome is an autoimmune disease that predominantly affects the salivary and lacrimal glands. Immune-mediated destruction of the lacrimal gland produces keratoconjunctivitis sicca that can be difficult to manage with conventional dry eye therapies. Prosthetic Replacement of the Ocular Surface Ecosystem (PROSE) therapy utilizes a custom-designed scleral device that vaults the entire cornea and limbus, bathing the ocular surface in preservative-free sterile saline. PROSE therapy has been shown to be beneficial in numerous other ocular surface diseases. The objective of this study was to evaluate the utility of PROSE therapy for patients with Sjogren’s related dry eye disease with outcomes based on visual acuity and function. Methods: The University of Southern California Institutional Review Board approved this study. A retrospective review from July 2009 to October 2015 of patients referred to USC Eye Institute for PROSE therapy consultation identified 37 patients with Sjogren’s syndrome related dry eye disease. Eight patients were excluded because they did not complete the fitting process for reasons such as being lost to follow-up, inability to obtain insurance authorization, difficulty with insertion and removal of the lens, or desire to continue with topical therapy. One additional patient was excluded because he was previously fit with PROSE at another institution. Of the 28 remaining patients, visual acuity, before and after PROSE fitting, was assessed using a Snellen chart under standardized conditions. Visual function These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts was assessed before and after PROSE fitting using the Ocular Surface Disease Index (OSDI) questionnaire. Wilcoxon signed-rank test was used to evaluate changes in visual acuity and OSDI scores. Results: Fifty-six eyes from 27 female patients and 1 male patient were included in this initial study. The average age was 52.3 years with a range of 30 to 65 years. The average visual acuity improved from 0.18 ± 0.20 logMAR (approximately 20/30) pre-PROSE to 0.08 ± 0.15 logMAR (approximately 20/25) post-PROSE (Z = -4.42; p < 0.01). Nine patients completed both the pre- and post-PROSE OSDI questionnaire. The average OSDI score improved from 66.9 ± 23.1 pre-PROSE to 31.4 ± 12.6 post-PROSE (Z = -3.24; p < 0.01). Conclusions: The data suggest that PROSE therapy is an effective treatment for patients suffering from Sjogren’s related dry eye disease Commercial Relationships: Billy X. Pan, None; Gloria B. Chiu, None; Martin Heur, None Program Number: 3891 Poster Board Number: A0039 Presentation Time: 3:45 PM–5:30 PM Prevalence and risk factors of superior limbal keratoconjunctivitis in Graves’ disease Anfal Almazrooei1, Omnia Hamam1, Laurence Dupasqiuer2, Michel Berche1, Emmanual Heron2, Christophe Baudouin1, Antoine Labbé1. 1Service d’Ophtalmologie, CHNO des QuinzeVingts, Paris, France; 2Department of Internal Medicinel, CHNO des Quinze-Vingts, Paris, France. Purpose: To evaluate the prevalence and risk factors of superior limbal keratoconjunctivits (SLK) in patients with Graves’ disease. Methods: Thirty-five patients diagnosed as having SLK from a cohort of 459 patients with Graves’ disease were retrospectively evaluated. SLK was defined as a superior ocular surface fluorescein staining associated with superior conjunctival laxity and inflammation of the superior tarsal and bulbar conjunctiva. Demographic data and clinical features were evaluated and all patients had a complete evaluation of the ocular surface. Graves’ orbitopathy was diagnosed on the basis of the criteria of the European Group On Graves’ orbitopathy (EUGOGO) and the disease activity was quantified according to the Clinical Activity Score (CAS) and Ocular Surface Disease Index (OSDI) questionnaires. Results: Within the 35 patients with SLK, 75.8% were female. Most of the patients were Caucasian (48.5%) followed by African (27,3%) and Asian (18,2%). Mean age was 45.48 ± 13.2 years. Considering smoking habit, 42.4% of patients never smoked while 39.4% were current smokers. Diabetes was found in 3.2% of patients. Bilateral SLK was observed in 54.8% of cases. Although there were good correlations between CAS score and antibodies against TSH receptor (TRAK) (p=0.03) and OSDI (p<0.05), there was no significant correlation between CAS score and the presence of SLK. Conclusions: SLK is not unusual in patient with Graves’ disease however it is not related to its severity. Careful ocular surface evaluation has to be performed in Graves’ orbitopathy to diagnose SLK. Commercial Relationships: Anfal Almazrooei, None; Omnia Hamam; Laurence Dupasqiuer, None; Michel Berche, None; Emmanual Heron, None; christophe baudouin, None; Antoine Labbé, None Program Number: 3892 Poster Board Number: A0040 Presentation Time: 3:45 PM–5:30 PM Modulating the toxic effects of benzalkonium chloride on human corneal epithelial cells in vitro Elham Ghahari, Mohammadreza Ghahari, Sanaz Gidfar, Behrad Y. Milani, Sara Sanjari, Medi Eslani, Thasarat S. Vajaranant, Ahmad A. Aref, Ali R. Djalilian. ROI, University of Illinois at Chicago, Chicago, IL. Purpose: Many topical ophthalmic medications have documented ocular surface toxicity attributed to preservatives, most commonly benzalkonium chloride (BAK). The main purpose of this study is to determine the critical cytotoxic BAK concentration for telomerizedimmortalized Human Corneal Epithelial Cells (HCECs) using a series of concentrations and different exposure time, and find a way to prevent or treat this damage. Methods: HCECs was plated in 96-well plates and incubated for 1 day. Culture medium was then replaced by basal media with different BAK concentrations (0.0005% to 0.005% in 0.0005 increment steps), incubated for 5, 10, or 15 minutes. In another set of experiments, the cells were incubated in a specific conditioned media 30 minutes before BAK application. Cell proliferation/viability was assessed by MTT assay. Results: BAK treatment time of 10 minutes was selected. BAK toxic concentration was 0.0015. Our results suggest that conditioned media has some degrees of protective effects against BAK ocular toxicity. HCECs pre-treated with the specific conditioned media demonstrated 71% cell viability in comparison to 56% for un-conditioned media. Conclusions: These results indicate that BAK induced cell damage might be reduced using a specified conditioned media. This may be used clinically to decrease the adverse effect of topical eye drops. Commercial Relationships: Elham Ghahari; Mohammadreza Ghahari, None; Sanaz Gidfar, None; Behrad Y. Milani, None; Sara Sanjari, None; Medi Eslani, None; Thasarat S. Vajaranant, None; Ahmad A. Aref, None; Ali R. Djalilian, None Program Number: 3893 Poster Board Number: A0041 Presentation Time: 3:45 PM–5:30 PM Corneal bioimpedance evaluation in an animal model of anterior surface inflammation Mario Crespo-Moral1, Alfredo Holgueras-López1, Anton Guimerà2, 3, Rosa Villa2, 3, Miguel Maldonado1. 1IOBA - University of Valladolid, Valladolid, Spain; 2Institut de Microelectrònica de Barcelona IMBCNM (CSIC), Barcelona, Spain; 3CIBER-BBN, Networking Center on Bioengineering, Biomaterials and Nanomedicine, Zaragoza, Spain. Purpose: Changes in the corneal barrier function can be shown by measuring the corneal bioimpedance. We evaluated if we were able to charactericise the damage produced in an experimental model of anterior surface inflammation (ASI) using a tetrapolar impedance meter. Methods: ASI was produced in 14 female New Zealand white rabbits by two different methods. In the first one (n=4) 40ul of 3% croton oil diluted in 2-ethoxyethanol was administered into the fornix of one of the eyes from each animal while the vehicle was administered into the other one. In the second one (n=10) 20ul of 3% croton oil diluted in dimethyl sulfoxide (DMSO) were administered into the fornix of one of the eyes from each animal while the vehicle was administered into the other one. Bioimpedance measures, mucous secretion (MS), chemosis (CH), neovascularization (NV) and corneal staining (CS) were assessed at each time point evaluation (baseline, 6, 24, 48 and 148 hours after exposure). Corneal tissue was extracted at the These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts endpoint, evaluated by light microscopy, and inmunostained for ZO-1 and ZO-2. Results: In the first group we show that MS, CH and CS were present in the 6, 24 and 48 hours evaluations in 3/4 of the cases while only MS and CS were present in the remaining case after 6 and 24 hours. In the second group we show MS in every animal after 6 hours and CH in 90% of the cases after 6 hours and 80% after 24 hours, CS is not present at any time point of the evaluation. Bioimpedance measures show low impedance values after 6 and 24 hours in the first group while there were no differences with the basal values at other times. In the second group higher impedance values were observed 6 and 24 hours after the administration in 70% of the cases while there were no differences at anyother time point. Conclusions: We had been able to reproduce a short time animal model of ASI and measure its bioimpedance at different time points. The combination of croton oil 3% in DMSO is the best way of the two methods to study the effect of inflammation over the ocular surface because it do not damage corneal epithelium integrity, even though it produces shorter time effects. The bioimpedance measures reflect damage in the corneal barrier function at the same times that corneal staining is observed. Commercial Relationships: Mario Crespo-Moral, None; Alfredo Holgueras-López, None; Anton Guimerà, Laboratorios SALVAT S.A. (P); Rosa Villa, Laboratorios SALVAT S.A. (P); Miguel Maldonado, Laboratorios SALVAT S.A. (P) Program Number: 3894 Poster Board Number: A0042 Presentation Time: 3:45 PM–5:30 PM Reproducibility and Outcomes of Simple Limbal Epithelial Transplantation (SLET) technique Alexandra Abdala, Arturo J. Ramirez-Miranda, Alejandro Lichtinger, Alejandro Navas, Enrique O. Graue-Hernandez. Instituto de Oftalmologia Conde de Valenciana, Mexico City, Mexico. Purpose: To report the results of simple limbal epithelial transplantation (SLET) for limbal stem cell deficiency (LSCD) at an ophthalmological institution in Mexico City. Methods: Five patients (5 eyes) with LSCD due to chemical injuries (3 eyes), thermal injury (1 eye) and failure of tectonic sclerokeratoplasty after corneal melting of tectonic patch graft secondary to rheumatoid arthritis perforation (1 eye). All patients had unilateral involvement and received an autologous graft of limbal stem cells from the contralateral healthy eye, technique described by Sangwan. Preoperative best-corrected visual acuity, postoperative best-corrected visual acuity, previous surgeries, side effects on the donor eye, time from injury to surgery, recurrence and postoperative complications were evaluated. Paired t test was used for statistical analysis. Results: Preoperative mean best-corrected visual acuity was 1.55LogMAR (1 to 2.3LogMAR). The mean follow-up was 10.2 months (6 to 14 months), with postoperative mean best-corrected visual acuity of 0.33LogMAR (0.17 to 0.47LogMAR). The ocular surface was improved in all patients, however focal recurrence with superficial corneal vascularization was observed on the affected eye. The donor eye did not have complications. Conclusions: The result of our study suggests that SLET is a safe and effective treatment option for the management of unilateral LSCD, improving the ocular surface for future corneal procedures. Commercial Relationships: Alexandra Abdala, None; Arturo J. Ramirez-Miranda, Carl Zeiss Meditec (C); Alejandro Lichtinger, None; Alejandro Navas, Alcon Laboratories Inc (C); Enrique O. Graue-Hernandez, None Program Number: 3895 Poster Board Number: A0043 Presentation Time: 3:45 PM–5:30 PM Topical Interferon Alpha-2b In Giant Squamous Ocular Surface Neoplasia Erick Hernandez-Bogantes, Andrew Olivo-Payne, Alexandra Abdala, Juan Carlos Serna-Ojeda, Enrique O. Graue-Hernandez, Alejandro Navas, Arturo J. Ramirez-Miranda. Cornea and Refractive Surgery, Insituto de Oftalmologia Fundacion Conde de Valenciana, Mexico D.F, Mexico. Purpose: To report a case series of 8 patients clinically diagnosed with giant ocular squamous neoplasia (OSSN) who underwent medical therapy with topical interferon alpha-2b for immunoreduction and immutherapy purposes Methods: Patients received monthly dosis of topical interferon alpha-2b (1 million IU/mL) for a total of 3 months or with a subconjunctival injection (3 million IU/mL) single dose per week monthly until tumor resolution. Results: A total of eight eyes of 8 patients (mean age of 69 y/o) with a clinical diagnosis of unilateral giant OSSN were managed with interferon alpha-2b. Six patients received topical therapy (1 million IU/mL) 4 times per day for a minimum of 3 months for immunotherapeutic purposes and 2 patients received subconjunctival therapy (3 million IU/mL) single dose per week for 1 month for immunoreduction. The median time to resolution was 5 months (range, 1.5-6 months) and the median follow-up was 22 months (range, 6-28 months). Despite the dramatic clinical response, all patients continued with topical interferon alpha-2b at least 1 month beyond complete clinical resolution. The two cases in which immunoreduction was successfully achieved, were scheduled for a lesion excision with a no-touch technique followed by cryotherapy and amniotic membrane to cover the large excision area. There has been no recurrence during follow-up. Conclusions: In cases of giant OSSN interferon alpha-2b can successfully reduce or achieve complete resolution, thus avoiding the patient a more extensive surgical procedure with its known risks and complications. These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record. ARVO 2016 Annual Meeting Abstracts Results: MSCs were isolated and their specific markers, such as CD29, CD44, CD105 and CD90, were expressed at high levels, while CD31, CD34, CD45 and MHC-DR could not be detected. Rat conjunctiva showed mild hyperemia at the injection site on the second day and diminished on day 3. All the injected cells remained in the engrafted regions and showed no migration. Majority of the MSCs survived 3 days after implantation. However, the number of cells gradually decreased thereafter, and completely disappeared on day 9 after implantation. Using Ki67 as a marker, we could only observe quite a number of positive staining cells surrounding the cell mass. Immunostaining against a-SMA was negative from day 3 to day 9, indicating that the injected cells did not differentiate into myofibroblasts. TUNEL positive cells were present in the injected cell mass. The quantities of infiltrated CD11b+, CD45+ and PMN+ cells were increased in the injected eyes at day 3, and gradually reduced to normal basal levels at day 9. Conclusions: MSCs were gradually eliminated from the injection site without cell migration, proliferation and differentiation. However, MSCs activated proliferation of host cells and recruited immune cells to the microenvironment. Commercial Relationships: Juan Li, None; Chengyou Zuo, None; Shangkun Ou, None; Sanming Li, None; Liying Zhang, None; Changkai Jia, None; Zuguo Liu, None; Wei Li, None Support: Chinese National Key Scientific Research Project (2013CB967003) Commercial Relationships: Erick Hernandez-Bogantes, None; Andrew Olivo-Payne, None; Alexandra Abdala, None; Juan Carlos Serna-Ojeda, None; Enrique O. Graue-Hernandez, None; Alejandro Navas, None; Arturo J. Ramirez-Miranda, None Program Number: 3896 Poster Board Number: A0044 Presentation Time: 3:45 PM–5:30 PM The fate of mesenchymal stem cells after subconjunctival implantation Juan Li, Chengyou Zuo, Shangkun Ou, Sanming Li, Liying Zhang, Changkai Jia, Zuguo Liu, Wei Li. Xiamen University Medical College, Eye Institute of Xiamen University, Xiamen, China. Purpose: Subconjunctival injection of mesenchymal stem cells (MSCs) has been applied in the treatment of ocular surface chemical burn, while the fate of MSCs in the local microenvironment after injection remains unknown. In this study, we investigated the survival, migration, proliferation, differentiation and paracrine of human MSCs, as well as their impact on the ocular surface microenvironment after subconjunctival implantation in rat. Methods: MSCs were isolated from umbilical cord characterized by flow cytometry on the basis of published criteria. Overall, 27 SD rats were subjected to MSCs injection. For each rat, the left eye was injected subconjunctivally with 2X105 MSCs suspended in 50ml DMEM, and the right eye was served as a control. On the 3rd, 6th and 9th day after injection, the rats were examined by slit-lamp to evaluate conjunctival edema and hyperemia. Immunohistochemistry was performed to investigate the expression patterns of Vimentin, a-SMA, Ki67 and PMN. TUNEL assay was performed to detect cell apoptosis. Furthermore, the proportion of CD11b+ and CD45+ cells in conjunctiva were quantified by flow cytometry. These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/ to access the versions of record.