Variants in IL23R-C1orf141 and ADO-ZNF365-EGR2 are associated with susceptibility to Vogt-Koyanagi-Harada disease in Japanese population.

Vogt-Koyanagi-Harada (VKH) disease is a systemic inflammatory disorder that affects pigment cell-containing organs such as the eye (e.g., chronic and/or recurrent granulomatous panuveitis). While the exact etiology and pathogenic mechanism of VKH disease are unclear, HLA-DR4 alleles have been documented to be strongly associated with VKH disease in various ethnic groups. Recently, a genome-wide association study (GWAS) found two new genetic risk factors (IL23R-C1orf141 and ADO-ZNF365-EGR2) in a non-HLA region from a Han Chinese population. In this study, we replicated these GWAS findings in a Japanese population. A total of 1,643 Japanese samples (380 cases with VKH disease and 1,263 healthy controls) were recruited. We assessed four single nucleotide polymorphisms (SNPs) shown in previous GWAS: rs78377598 and rs117633859 in IL23R-C1orf141, and rs442309 and rs224058 in ADO-ZNF365-EGR2. A significant allelic association with VKH disease was observed for all of the four SNPs (rs78377598: pc = 0.0057; rs117633859: pc = 0.0017; rs442309: pc = 0.021; rs224058: pc = 0.035). In genotypic association analysis, the minor alleles of IL23R-C1orf141 rs78377598 and rs117633859 had the strongest association with disease susceptibility under the additive model (pc = 0.0075 and pc = 0.0026, respectively). The minor alleles of ADO-ZNF365-EGR2 rs442309 and rs224058 were most strongly associated with disease susceptibility under the dominant model (pc = 0.00099 and pc = 0.0023, respectively). The meta-analysis of the current and previous studies found that all of the four SNPs exhibited a significantly strong association with VKH disease (meta-p < 0.00001: rs78377598, meta-odds ratio (OR) = 1.69; rs1176338, meta-OR = 1.82; rs442309, meta-OR = 1.34; rs224058, meta-OR = 1.33). In summary, our study replicated significant associations with VKH disease susceptibility reported in a previous GWAS. Thus, the IL23R-C1orf141 and ADO-ZNF365-EGR2 loci may play important roles in the development of VKH disease through genetic polymorphisms.

Relationship between Vision-Related Quality of Life and Central 10° of the Binocular Integrated Visual Field in Advanced Glaucoma.

To investigate the relationships between sensitivity loss in various subfields of the central 10° of the binocular integrated visual field (IVF) and vision-related quality of life (VRQoL) in 172 patients with advanced glaucoma. Using the Random Forest algorithm, which controls for inter-correlations among various subfields of the IVF, we analysed the relationships among the Rasch analysis-derived person ability index (RADPAI), age, best-corrected visual acuity (BCVA), mean total deviations (mTDs) of eight quadrant subfields in the IVF measured with the Humphrey Field Analyzer (HFA) 10-2 program (10-2 IVF), and mTDs of the upper/lower hemifields in the IVF measured with the HFA 24-2 program (24-2 IVF). Significant contributors to RADPAIs were as follows: the inner and outer lower-right quadrants of the 10-2 IVF contributed to the dining and total tasks; the lower-left quadrant of the 10-2 IVF contributed to the walking, going out and total tasks; the lower hemifield of the 24-2 IVF contributed to the walking, going out, dining, miscellaneous and total tasks; and BCVA contributed more to the letter, sentence, dressing and miscellaneous tasks than to others. The impact of damage in different 10-2 IVF subfields differed significantly across daily tasks in patients with advanced glaucoma.

Association Study of ARMC9 Gene Variants with Vogt-Koyanagi-Harada Disease in Japanese Patients.

Purpose: To investigate whether variants in the ARMC9 gene encoding KU-MEL-1 are associated with Vogt-Koyanagi-Harada (VKH) disease in a Japanese population. Methods: We recruited 380 Japanese patients with VKH disease and 744 Japanese healthy controls to genotype seven single-nucleotide polymorphisms (SNPs) in ARMC9. We also performed imputation analysis of the ARMC9 region and 195 imputed SNPs were included in the statistical analysis. Results: We observed an increased frequency of the A allele of rs28690417 in patients compared with controls (P = 0.0097, odds ratio (OR) = 1.46). The A allele had a dominant effect on VKH disease risk (P = 0.011, OR = 1.51). However, these significant differences disappeared after Bonferroni correction (corrected P > 0.05). The remaining 201 SNPs did not show any significant association with disease risk. Conclusions: Our study suggests that ARMC9 variants do not play a critical role in the development of VKH disease.

Long-term outcomes and prognostic factors for trabeculectomy with mitomycin C in eyes with uveitic glaucoma: a retrospective cohort study.

PURPOSE: To elucidate the long-term outcomes and prognostic factors for trabeculectomy with mitomycin C (MMC) in eyes with uveitic glaucoma (UG). METHODS: A retrospective, consecutive, comparative cohort study was conducted with 204 patients who underwent trabeculectomy with MMC between 1999 and 2008 at 2 Japanese clinical centers. The study group included 101 eyes with UG and 103 eyes with primary open-angle glaucoma (POAG). Surgical failure was defined as intraocular pressure levels of ≥21 mm Hg or an additional glaucoma surgery. Kaplan-Meier survival curves for surgical failure were compared between UG and POAG eyes, and prognostic factors for surgical failure of trabeculectomy in UG eyes were analyzed by the Cox proportional hazards model. Secondary outcome measures included comparisons of the frequency of additional cataract surgery and other surgical complications after trabeculectomy between UG and POAG eyes. RESULTS: The mean follow-up periods (±SD) were 34.7±37.9 and 37.7±34.7 months (median, 24.0 and 27.4 mo) for UG and POAG, respectively. The subtypes of uveitis were granulomatous uveitis (n=20) including sarcoidosis (n=12), Vogt-Koyanagi-Harada disease (n=5) and varicella zoster virus uveitis (n=3), Behçet disease (n=10), Posner-Schlossman syndrome (n=5), and other types of UG (n=12). Fifty-four eyes were diagnosed with idiopathic UG. The 3-year probabilities of success after trabeculectomy were 71.3% and 89.7% for UG and POAG, respectively (P=0.0171). A multivariable model showed that UG eyes with previous cataract surgery [relative risk (RR)=2.957, P=0.0344)] and granulomatous uveitis (RR=3.805, P=0.0106) were associated with surgical failure. UG eyes experienced more frequent cataract surgeries after trabeculectomy than POAG eyes: the 3-year probabilities of additional cataract surgery of 62.6% and 10.7% for UG and POAG, respectively (P<0.0001). There was no significant difference in the frequency of surgical complications such as bleb leakage, hypotensive maculopathy, severe anterior-chamber hemorrhage, and infectious endophthalmitis. CONCLUSIONS: Trabeculectomy with MMC was less effective in maintaining intraocular pressure reduction in UG eyes than in POAG eyes. The prognostic factors for surgical failure of trabeculectomy in UG eyes were previous cataract surgery and granulomatous uveitis. In addition, UG eyes after trabeculectomy more frequently required additional cataract surgery.

Feasibility of using gelatin-microbial transglutaminase complex to repair experimental retinal detachment in rabbit eyes.

BACKGROUND: To investigate the effect of using gelatin-microbial transglutaminase (gelatin-mTG) complex for treating experimental retinal detachment. METHODS: Vitrectomy with artificial posterior vitreous detachment (PVD) followed by induction of a retinal tear and detachment was performed in rabbit eyes. Gelatin-mTG complex or gelatin alone (control) was placed on the retinal tears. Fundus examination using optical coherence tomography (OCT) was performed after the surgery. Vitrectomy with PVD alone was also performed in additional rabbits. After application of the gelatin-mTG complex on the normal retinal surface, the electroretinogram (ERG) was measured 7 days after surgery. RESULTS: Gelatin-mTG complex covered the retinal tear for more than 7 days after the vitrectomy, with less prominent inflammation. Reattachment of the retina occurred in all treated eyes. In contrast, massive fibrin materials were observed at 1 day after the surgery in the control group. In addition, OCT showed that all of the gelatin disappeared by day 3. Local retinal detachment remained in three of the eyes. As demonstrated by the ERG, gelatin-mTG complex had no harmful effects on retinal function. CONCLUSIONS: The results indicate that gelatin-mTG complex continues to adhere and seal retinal tears for at least several days after administration without any inflammatory reaction.

A simulated eye for vitreous surgery using Japanese quail eggs.

PURPOSE: We produced a simulated eye for vitreous surgery training by using Japanese quail eggs, and verified its utility. METHODS: We used a special cutter to cut off the sharp end of a Japanese quail egg, fitted a silicone simulated sclerocorneal cap to the exposed area, and fixed the egg to a base. Trocars were placed in the simulated sclera according to the usual procedure for vitreous surgery, and the yolk and albumen were treated as the vitreous body, and resected by using a vitreous cutter. Membrane peeling was performed on the inner eggshell membrane as if it were the internal limiting membrane. RESULTS: The yolk and albumen could be resected with a vitreous cutter in the same way as the vitreous body. The inner eggshell membrane could be visualized under staining with Brilliant Blue G and other stains, enabling peeling to be performed with vitreous forceps in the same way as is normally performed for the human internal limiting membrane. CONCLUSION: This model can be used for simulating the spatial recognition of the vitreous chamber during vitreous surgery. This model proved useful for initial training in port creation, central vitreous body resection, and membrane manipulation in the macular area.

Use of an ophthalmic viscosurgical device for experimental retinal detachment in rabbit eyes.

To investigate the temporary tamponade effects of an ophthalmic viscosurgical device (OVD) for experimental retinal tears, we performed vitrectomy in four rabbit eyes and created a posterior vitreous detachment and artificial retinal tear to produce retinal detachment. The retina was flattened with liquid perfluorocarbon (PFC), the area peripheral to the tear was photocoagulated, an OVD was applied to the retinal tear surface below the PFC and the PFC was removed by aspiration. In the control group, PFC was removed without application of OVD. At one, three and seven days postoperatively, funduscopy and optical coherence tomography (OCT) were performed to examine the sealing process of the retinal tear. In OVD-treated eyes, the OVD remained on the retinal surface, and the retinal tear was patched for ≥ 3 days postoperatively. By seven days postoperatively, the OVD on the retinal surface had disappeared, and the retina was reattached. In control eyes, the edge of the retinal tear was rolled, and retinal detachment persisted. In OVD-treated eyes, the border of the retinal tear was indistinct, and the defect area was significantly decreased. These results show that application of an OVD effectively seals retinal tears and eliminates retinal detachments.

Observation of peripheral retina by topical endoscopic imaging method-a preliminary study.

INTRODUCTION: This study evaluated the usefulness of rigid endoscopy placed on the corneal surface to observe the peripheral retina. METHODS: The authors studied 15 eyes in 15 patients (12 men, 3 women; mean age 55.9 years; range 22-74 years) that underwent vitreous surgery at the Department of Ophthalmology at Saga University Hospital. With patients in a supine position, after topical anesthesia, an eye cup was placed between the eyelids and filled with hydroxyethyl cellulose solution and physiologic saline. With a rigid endoscope placed near the corneal surface, the target areas were then observed and recorded. The usefulness of rigid endoscopy to observe the peripheral retina was evaluated based on differences due to lens status and pupil size. RESULTS: In seven aphakic eyes, irrespective of pupil size, the peripheral retina could be observed up to the entire ora serrata (all quadrants). In eight eyes implanted with an intraocular lens, the observable area changed with pupil size and anterior capsulorrhexis size. CONCLUSION: This technique using rigid endoscopy was simple to manipulate and useful for observing and recording the peripheral retina. In particular, in aphakic eyes, irrespective of pupil size, the retina could be observed to the ora serrata.

Viability of topical endoscopic imaging system for vitreous surgery in rabbit eyes.

Vitreous surgery was performed in four rabbit eyes to investigate the viability of a topical endoscopic imaging system. This technique was evaluated during core vitrectomy, artificial posterior vitreous detachment, peripheral vitrectomy, production of intentional retinal breaks, and endophotocoagulation. To observe the posterior fundus during the surgeries, the endoscope was fixed in place. In addition, by tilting the endoscope toward the ciliary body, it was possible to clearly observe the trocar ports. The endoscope was easy to use and obtained good resolution. In addition, all procedures could be performed accurately when using the technique. The current study demonstrated that the topical endoscopic imaging system is a convenient method for obtaining both wide-field viewing and panfocal imaging during vitrectomy. This method may prove to be a new observational tool that can be employed during intraocular surgeries in the clinical setting.

Neuroprotective effect of small interfering RNA targeted to caspase-3 on rat retinal ganglion cell loss induced by ischemia and reperfusion injury.

PURPOSE: To investigate neuroprotective effects of siRNA targeted to caspase-3 against ischemia and reperfusion (I/R) injury in rat eyes. METHODS: Retinal ischemia was induced in Wistar rats by increasing the intraocular pressure (IOP) to 110 mmHg for 120 min. To examine the effect of siRNA on rat caspase-3, siRNA was injected into the vitreous cavity 24 h prior to induction of retinal ischemia. Eyes were removed at 2, 7 or 14 days later, and then analyzed for the number of retinal ganglion cells (RGCs), the retinal thickness and the amount of apoptosis of the retinal neural cells (as demonstrated by the TUNEL assay). The amount of caspase-3 mRNA was analyzed by rt-PCR. Differences between groups were evaluated by an unpaired t test. RESULTS: The numbers of RGCs in the saline and non-silencing siRNA controls were reduced significantly at 2 and 7 days after the I/R injury. RGCs were significantly retained in eyes pretreated with siRNA targeted to caspase-3 as compared to the control eyes at 2 days after the I/R injury. Inner retinal thickness in the control eyes was significantly thinner as compared to the treated eyes at 2 and 7 days after the I/R injury. After siRNA treatment, the amount of caspase-3 mRNA was significantly lower when compared to the saline control group. CONCLUSIONS: The injection of siRNA targeted to caspase-3 into the vitreous cavity of rat eyes may block caspase-3, and may thus be able to prevent retinal cell death associated with ischemic injury. As inhibition of the apoptosis pathway may provide a neuroprotective effect, examination of new strategies for treating these disorders needs to be undertaken.

Relapse of Vogt-Koyanagi-Harada Disease during Interferon-α and Ribavirin Therapy in a Case of Chronic Viral Hepatitis C.

A 60-year-old woman, who had been diagnosed with and treated for Vogt-Koyanagi-Harada (VKH) disease 17 years before, was receiving pegylated interferon-? and ribavirin therapy for chronic hepatitis C virus. Three weeks after the start of therapy, she complained of visual blurring, eye pain, and an increased hearing loss. Based on a slit lamp and fundus examination, she was diagnosed with a relapse of VKH disease. After discontinuation of the pegylated interferon-α and ribavirin therapy and administration of corticosteroid therapy, her visual acuity returned to 1.0 in both eyes without ocular inflammation, and remained stable thereafter. When interferon therapy is administered to hepatitis C virus patients, those who also have a history of VKH disease must be closely monitored for ophthalmologic complications.

Occurrence of capsular delamination in the dislocated in-the-bag intraocular lens.

PURPOSE: To examine the lens capsules of dislocated in-the-bag IOLs. METHODS: Retrospective case series. Nineteen dislocated IOLs encased in capsules (in-the-bag IOLs) from 19 patients were included. The clinical characteristics of the 19 patients were reviewed. The explanted in-the- bag IOLs were examined by scanning electron microscopy. RESULTS: Associated clinical conditions included pseudoexfoliation (PEX) in seven eyes, high myopia in three eyes, previous history of trauma in two eyes, previous vitreoretinal surgery in two eyes, retinitis pigmentosa in one eye and uveitis in one eye. PEX specimens showed capsular contraction, shrinkage of the diameter of the capsular bag and dehiscence of zonular fibers. The remaining 12 specimens exhibited slight capsular contraction that lacked shrinkage of the bag and exhibited capsular delamination at the equatorial region, in which the zonular fibers had completely disappeared. CONCLUSION: Capsular delamination as well as dehiscence of zonular fibers may be involved in the dislocation of in-the-bag IOLs.

[Evaluation of revised diagnosis criteria for sarcoidosis].

PURPOSE: Based on diagnostic criteria revised in 2006, we investigate whether a simple examination will provide a diagnosis for uveitis of sarcoidosis. SUBJECTS AND METHODS: Two hundred and six patients with uveitis suspected of having sarcoidosis who visited 4 hospitals from 1978 to 2008 were evaluated according to Their ocular and systemic findings. RESULTS: One hundred and six patients were diagnosed as having sarcoidosis. Most patients had more than 4 ocular criteria. Bilateral hilar lymphadenopathy (BHL) was seen in 103 (97.2%) of the patients. When BHL was not detected with conventional chest X-rays, chest computerized tomography (CT) was useful in detecting BHL, especially in patients who were tuberculin negative. CONCLUSIONS: When sarcoidosis is suspected from ocular findings, chest X-rays, a tuberculin skin test and serum angiotensin converting enzyme should be performed first. Even if BHL is not detected, a chest CT is useful in cases that are tuberculin negative.

Heparan sulfate deficiency in periocular mesenchyme causes microphthalmia and ciliary body dysgenesis.

The heparan sulfate (HS) is a component of proteoglycans in the extracellular matrix and on cell surfaces, modulating developmental processes. The aim of this study is to investigate whether the defect of HS in the periocular mesenchyme impairs ocular morphogenesis. First, using Protein 0-Cre transgenic mice, we ablated Ext1, which encodes an indispensable enzyme for HS synthesis, in the developing periocular mesenchyme. The expression of Ext1 messenger RNA (mRNA) and HS were observed by RT-PCR and immunohistochemistry, respectively. The phenotypes in the mutant were evaluated by light microscopy and immunohistochemistry for cellular makers. Second, the distribution of the mutant periocular mesenchymal cells was tracked using a Rosa26 Cre-reporter gene. No mutant embryos (Protein 0-Cre;Ext1(flox/flox)) were identified after embryonic day 14.5 (E14.5). RT-PCR showed that an intense band amplified from Ext1 was observed in cDNAs from the control periocular mesenchymal cells at E13.5; however, the band for Ext1 was hardly detectable in cDNA from the mutant embryo, indicating that the mRNA was missing in the mutant periocular mesenchyme at E13.5. The HS expression was disrupted in the periocular mesenchyme of the mutant ocular tissues. The HS deficiency resulted in microphthalmia with reduced axial lengths, lens diameters, and vitreous sizes compared with the littermate eyes. The mutant embryos showed agenesis of the anterior chamber, where cells expressing Cre recombinase were distributed. Moreover, the mutants showed phenotypic alterations in the neural ectoderm including dysgenesis of the presumptive ciliary body and agenesis of the optic nerve head. These findings demonstrate that HS in the periocular mesenchyme plays a critical role in normal ocular morphogenesis, indicating reciprocal interactions between the periocular mesenchyme and the neural ectoderm.

Heparan sulfate deficiency leads to Peters anomaly in mice by disturbing neural crest TGF-beta2 signaling.

During human embryogenesis, neural crest cells migrate to the anterior chamber of the eye and then differentiate into the inner layers of the cornea, the iridocorneal angle, and the anterior portion of the iris. When proper development does not occur, this causes iridocorneal angle dysgenesis and intraocular pressure (IOP) elevation, which ultimately results in developmental glaucoma. Here, we show that heparan sulfate (HS) deficiency in mouse neural crest cells causes anterior chamber dysgenesis, including corneal endothelium defects, corneal stroma hypoplasia, and iridocorneal angle dysgenesis. These dysfunctions are phenotypes of the human developmental glaucoma, Peters anomaly. In the neural crest cells of mice embryos, disruption of the gene encoding exostosin 1 (Ext1), which is an indispensable enzyme for HS synthesis, resulted in disturbed TGF-beta2 signaling. This led to reduced phosphorylation of Smad2 and downregulated expression of forkhead box C1 (Foxc1) and paired-like homeodomain transcription factor 2 (Pitx2), transcription factors that have been identified as the causative genes for developmental glaucoma. Furthermore, impaired interactions between HS and TGF-beta2 induced developmental glaucoma, which was manifested as an IOP elevation caused by iridocorneal angle dysgenesis. These findings suggest that HS is necessary for neural crest cells to form the anterior chamber via TGF-beta2 signaling. Disturbances of HS synthesis might therefore contribute to the pathology of developmental glaucoma.

Changes in choriocapillaris fenestration of rat eyes after intravitreal bevacizumab injection.

BACKGROUND: To examine the effects of anti-VEGF antibody (bevacizumab) on the number of fenestrations in rat choriocapillaris. METHODS: Twenty-four eyes from 24 male Wister rats were injected intravitreally with 0.125 mg of bevacizumab. The rats were perfusion fixated at 1, 3, 7, 14 or 28 days after injection. The surfaces of the choriocapillaris on the RPE side were observed using scanning electron microscopy. Four eyes treated with human IgG were used as controls. The area sieve plate and the number of fenestrations after the bevacizumab injection were measured and compared with controls. RESULTS: In the controls, the sieve plate area was 80.7% of the total choriocapillaris area. The number of fenestrations was 69.2 +/- 0.2 /microm(2) of the fenestrated area. While there were no changes in the fenestrated area for any of the time points after the bevacizumab treatment, the number of fenestrations was significantly reduced to 52.9 +/- 4.4 at day 1, 55.6 +/- 3.6 at day 3 and 53.6 +/- 8.6 /microm(2) of the luminal surface at day 7 (ANOVA, p < 0.05). CONCLUSIONS: In this study, intravitreal bevacizumab injection reduced fenestration of the normal choriocapillaris. These results indicate there is a latent risk inherent with bevacizumab treatment of normal choriocapillaris.

Fluorescein and indocyanine green angiographic findings in progressive hemifacial atrophy.

A 20-year-old man who had progressive hemifacial atrophy was examined using fluorescein and indocyanine green angiography. The fundus showed sectional chorioretinal atrophy. Fluorescein angiography showed window defects without leakage. Indocyanine green angiography revealed narrow choroidal vessels with hypofluorescence.