Woo Jin Jeong

Cytoplasmic and Nuclear Anti-Apoptotic Roles of αB-Crystallin in Retinal Pigment Epithelial Cells

In addition to its well-characterized role in the lens, αB-crystallin performs other functions. Methylglyoxal (MGO) can alter the function of the basement membrane of retinal pigment epithelial (RPE) cells. Thus, if MGO is not efficiently detoxified, it can induce adverse reactions in RPE cells. In this study, we examined the mechanisms underlying the anti-apoptotic activity of αB-crystallin in the human retinal pigment epithelial cell line ARPE-19 following MGO treatment using various assays, including nuclear staining, flow cytometry, DNA electrophoresis, pulse field gel electrophoresis, western blot analysis, confocal microscopy and co-immunoprecipitation assays. To directly assess the role of phosphorylation of αB-crystallin, we used site-directed mutagenesis to convert relevant serine residues to alanine residues. Using these techniques, we demonstrated that MGO induces apoptosis in ARPE-19 cells. Silencing αB-crystallin sensitized ARPE-19 cells to MGO-induced apoptosis, indicating that αB-crystallin protects ARPE-19 cells from MGO-induced apoptosis. Furthermore, we found that αB-crystallin interacts with the caspase subtypes, caspase-2L, -2S, -3, -4, -7, -8, -9 and -12 in untreated control ARPE-19 cells and that MGO treatment caused the dissociation of these caspase subtypes from αB-crystallin; transfection of S19A, S45A or S59A mutants caused the depletion of αB-crystallin from the nuclei of untreated control RPE cells leading to the release of caspase subtypes. Additionally, transfection of these mutants enhanced MGO-induced apoptosis in ARPE-19 cells, indicating that phosphorylation of nuclear αB-crystallin on serine residues 19, 45 and 59 plays a pivotal role in preventing apoptosis in ARPE-19 cells. Taken together, these results suggest that αB-crystallin prevents caspase activation by physically interacting with caspase subtypes in the cytoplasm and nucleus, thereby protecting RPE cells from MGO-induced apoptosis.

Canaliculitis with a papilloma-like mass caused by a temporary punctal plug.

Absorbable plugs are used before attempting permanent occlusion to observe whether the patient will be amenable to treatment for dry eye. Little is known about the complications of absorbable plugs. The authors present a case of canaliculitis with a papilloma-like mass caused by long-standing unabsorbed collagen plug that had been placed 3 years earlier. This is an uncommon clinical presentation that may be encountered in patients who have canaliculitis symptoms and a history of intracanalicular temporary plug placement.

Sensitization of RPE Cells by αB-Crystallin siRNA to SAHA-Induced Stage 1 Apoptosis through Abolishing the Association of αB-Crystallin with HDAC1 in SC35 Speckles

PURPOSE To better understand the mechanism underlying the anti-apoptotic activity of alphaB-crystallin in RPE cells. METHODS Cells of the human retinal pigment epithelial line ARPE-19 were treated with a histone deacetylase inhibitor (HDACI), suberoylanilide hydroxamic acid (SAHA), with or without alphaB-crystallin siRNA. To examine the mechanism underlying the cell death induced in ARPE-19 cells, nuclear staining, flow cytometry, DNA electrophoresis, pulse field gel electrophoresis, Western blot analysis, confocal microscopy, and coimmunoprecipitation assay were undertaken. RESULTS The present study demonstrated that an HDACI, SAHA, at the usual doses or the silencing of alphaB-crystallin by siRNA alone did not effectively induce apoptosis in ARPE-19 cells. Silencing of alphaB-crystallin likely abolishes the anti-apoptotic activity of alphaB-crystallin. The data indicated that silencing of alphaB-crystallin sensitizes ARPE19 cells to SAHA-induced apoptosis and leads them to stage 1 apoptosis. alphaB-Crystallin associates with HDAC1 on SC35 speckles, and silencing of alphaB-crystallin abolishes this association, resulting in the induction of apoptosis. The data indicated that the association between alphaB-crystallin and HDAC1 on SC35 speckles plays a pivotal role in anti-apoptotic activity. CONCLUSIONS Knockout of alphaB-crystallin may be a promising new approach to enhance therapeutic potency for proliferative vitreoretinopathy without compromising efficacy.

The Clinical Characteristics and Surgical Outcomes of Epiblepharon in Korean Children: A 9-Year Experience

Purpose. To examine the demographic characteristics, clinical features, surgical outcomes, and long-term prognoses of epiblepharon in Korean children. Methods. Epiblepharon patients who were followed for ≥ 6 month following surgical correction between January 2005 and December 2013. The patient demographics, clinical features, concomitant disorders, surgical outcomes, and complications were retrospectively reviewed. Results. A total of 768 epiblepharon records were included in the analysis. The mean patient age was 6.55 ± 2.37 years. At presentation, 712 patients (92.8%) complained of typical epiblepharon symptoms. The mean patient age at surgery was 6.95 ± 2.52 years, with 629 patients (81.9%) on the lower lid and 72 patients (9.4%) on the upper lid and 82 patients (10.7%) undergoing surgery on both lids. The eyelid was well everted with no recurrence in 740 patients (96.4%). Conclusion. Epiblepharon frequently occurs in Korean children and is correctable with a simple surgery. Recurrence and serious complications do not occur often, and any suspicions of epiblepharon should be investigated. A thorough ocular examination can lead to a correct diagnosis and timely corrective surgery. Most procedures are successful and prevent secondary complications that often occur with uncorrected epiblepharon.

Epiblepharon Related to High Body Mass Index in Korean Children

PURPOSE To evaluate the clinical manifestations of epiblepharon in Korean children and the relationship between this condition and body mass index. METHODS In a retrospective case-control study, 223 children 6 to 15 years of age treated surgically for epiblepharon were studied and compared with an age-matched control group of 264 children. Data on age, sex, weight, height, and recurrence were recorded. RESULTS Mean age of the patients was 8.63 ± 4.43 years. Lower eyelids were commonly involved bilaterally. Chief complaints included ocular discomfort, eye rubbing, visual disturbances, photophobia, and epiphora. Minimal resection of orbicularis oculi with full-thickness eyelid rotatory suture for epiblepharon was performed on 223 patients. The body mass index in girls 12 to 15 years old with epiblepharon was significantly higher than that in children without epiblepharon (P < .05). There was a statistically significant correlation between the incidence of epiblepharon and obesity with aging in girls (P < .05). Reoperation was performed on 12 patients (4.4%) because of recurrence of corneal irritation with inverted eyelashes. Clinical factors such as sex, age, and body mass index did not affect the rate of recurrence. CONCLUSION Epiblepharon is one of the most common eyelid abnormalities among Asian children. Obese Korean girls aged 12 to 15 years demonstrated symptomatic epiblepharon at a statistically significant level.

Co-treatment of suberoylanilide hydroxamic acid and mitomycin-C induces the apoptosis of rabbit tenon's capsule fibroblast and improves the outcome of glaucoma filtration surgery.

Purpose: This study was undertaken to develop a new treatment modality that would be able to minimize fibrosis and provide better outcome with glaucoma filtration surgery (GFS). Methods: We examined whether co-treatment with mitomycin-C (MMC) and histone deacetylase inhibitor (HDACi) suberoylanilide hydroxamic acid (SAHA) efficiently induces apoptosis on rabbit Tenons capsule fibroblasts (TCF) in vitro. We further examined the effect of co-treatment with SAHA and MMC on the alteration of IOP and the bleb survival in rabbits following GFS. Results: Co-treatment of MMC and SAHA efficiently induces apoptosis in TCFs via the up-regulation of p53 and increased phosphorylation of p53 on serine 15 and 392. Also, co-treatment of SAHA and low-dose MMC decreases IOP, prolongs bleb survival, and induces apoptosis of cells under the bleb area following GFS. Conclusion: This study shows that a co-treatment of SAHA and MMC could improve the outcome of GFS.

Classification of Lacrimal Punctal Stenosis and Its Related Histopathological Feature in Patients with Epiphora

Purpose To evaluate the classification of punctal stenosis based on the shape of the external punctum, clinical characteristics and histopathologic features. Methods Patients who experienced tearing and were diagnosed with punctal stenosis were evaluated in this study. Punctal stenosis was classified according to the shape of the lower external punctum, which included membranous type, slit type, horseshoe type, and pinpoint type. Tear meniscus height, 2% fluorescein dye disappearance test and lacrimal pathway irrigation were measured or performed. For treatment, a punctal snip operation and silicone tube placement were performed, and the peripunctal histopathological findings were evaluated. Results Punctal stenosis was classified into four types: membranous type (17 eyes, 21.5%), slit type (11 eyes, 13.9%), horseshoe type (25 eyes, 31.6%), and pinpoint type (26 eyes, 32.9%). The tear meniscus was significantly higher, and the 2% fluorescein dye disappeared significantly more slowly in the punctal stenosis group. However, correlation of the tear meniscus height and 2% fluorescein dye disappearance test with the punctum shape was not statistically significant. A history of previous chemotherapy was significantly associated with the occurrence of punctal stenosis, especially the membranous type (p < 0.05). Histopathologic evaluation of the punctum showed differences between the punctum types. Pinpoint puncta exhibited a high density of muscle fibers, while they were faintly visible in the membranous type. Conclusions Acquired punctal stenosis has various shapes, and the major types of stenotic puncta exhibited unique histopathologic features. Punctal stenosis and its pathophysiology may be related to multiple factors, such as age and systemic 5-fluorouracil chemotherapy history.

Effects of upper lid blepharoplasty on visual quality in patients with lash ptosis and dermatochalasis

AIM To assess the effects of upper lid blepharoplasty on visual quality. METHODS Seventy-three eyelids of 39 patients were subjected to upper lid blepharoplasty. Pre- and post-operative contrast sensitivity, visual acuity, vertical palpebral aperture and the degree of lash ptosis were measured. RESULTS The mean log contrast sensitivities under photopic (P=0.017) and scotopic conditions (P=0.009) were improved after surgery, and these differences were significant. The degree of lash ptosis was also decreased after blepharoplasty (P<0.001). CONCLUSION In our study, a significant increase in contrast sensitivity was found after surgery. These results suggest that upper lid blepharoplasty can be helpful for improving visual quality.