Details of a Researcher - Tsubota, Kazuo

Papers - Tsubota, Kazuo

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Motor function benefits of visual restoration measured in age-related cataract and simulated patients

Ayaki Masahiko, Nagura Takeo, Toyama Yoshiaki, Negishi Kazuno, Tsubota Kazuo

Scientific Reports 5 2015.09

The aim of the present study was to measure gait velocity in cataract and simulated patients. The study was performed on 239 cataract patients, 115 age-matched subjects, and 11 simulated patients. We measured gait velocity and analyzed gait using a three-dimensional motion analysis system. Mean gait velocity before and 2 and 7 months after cataract surgery was 0.91 ± 0.19, 1.04 ± 0.21, and 1.06 ± 0.21 m/s, respectively, for males and 0.84 ± 0.22, 0.91 ± 0.24, and 0.92 ± 0.25 m/s, respectively, for females. The increase after surgery was significant in both groups at 7 months (P < 0.05). Gait velocity was significantly slower in cataract patients compared with controls before surgery, but no longer different after surgery. In simulated patients, mean velocity was 87.0 ± 11.4% of normal vision with a 3° visual field and 92.4 ± 12.3% of normal when counting fingers. Initial velocity was 89.1 ± 14.6% of normal vision with a 3° visual field and 92.7 ± 11.6% of normal when counting fingers. There was a significant difference between normal and impaired visual function (P < 0.05). The results demonstrate the close relationship between visual function and gait in cataract patients and simulated patients.

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Possible involvement of epithelial-mesenchymal transition in fibrosis associated with IgG4-related Mikulicz's disease

Fukui Masaki, Ogawa Yoko, Shimmura Shigeto, Hatou Shin, Ichihashi Yoshiyuki, Yaguchi Saori, Hirayama Masatoshi, Kawakita Tetsuya, Tsubota Kazuo

Modern Rheumatology 25 ( 5 ) 737 - 743 2015.09

ISSN 1439-7595

Objective. Immunoglobulin G4 (IgG4)-related Mikulicz's disease (MD) is a fibrosis-associated inflammatory disease, often accompanied by lacrimal gland swelling. Although much attention has been paid to the inflammatory aspects of this disease, the mechanisms of the fibrotic processes are still unclear. We focused on the fibrotic changes occurring in the lacrimal glands of IgG4-related MD patients, by examining molecules involved in the epithelial-mesenchymal transition (EMT). Methods. Lacrimal gland tissue specimens were obtained from 3 IgG4-related MD patients and 3 control patients with Sjögren's syndrome (SS). The glands were examined by immunohistochemistry and transmission electron microscopy. Results. Storiform fibrosis, a characteristic of IgG4-related MD, was observed in the lacrimal glands of IgG4-related MD, but rarely in those of SS. Reduced E-cadherin expression, increased phalloidin-stained filamentous actin, and increased α-smooth muscle actin, snail, and heat-shock protein 47 levels were observed in the lacrimal glands of IgG4-related MD compared with those of SS. Transmission electron microscopy revealed an abnormal periodicity of collagen bundles, and basal membrane thickening in the IgG4-related MD compared with that in the SS tissues. Conclusion. EMT-like changes were frequently observed in the lacrimal gland epithelia from patients with IgG4-related MD. Thus, EMT may be involved in the pathology of IgG4-related MD fibrosis.

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Wide-Angle Viewing System versus Conventional Indirect Ophthalmoscopy for Scleral Buckling

Tomita Yohei, Kurihara Toshihide, Uchida Atsuro, Nagai Norihiro, Shinoda Hajime, Tsubota Kazuo, Ozawa Yoko

Scientific Reports 5 2015.09

Wide-angle viewing systems (WAVSs) were originally established for pars plana vitrectomy. However, their application to scleral buckling surgery was recently reported. In this study, we compared the outcomes of scleral buckling using a noncontact WAVS with that of scleral buckling using conventional indirect ophthalmoscopy for rhegmatogenous retinal detachment. The clinical records of 39 eyes (39 patients) with rhegmatogenous retinal detachment primarily treated between November 2012 and June 2014 at the Vitreo-Retina Surgical Division Clinic at the Department of Ophthalmology, Keio University Hospital were retrospectively reviewed. Scleral bucking was performed using WAVS with surgical placement of an endoilluminator in 16 eyes and indirect ophthalmoscopy in 23 eyes. The patients in these groups were consecutive over different intervals. The preoperative demographics, success rate of retinal reattachment, intraoperative findings, and postoperative complications were evaluated. There were no significant differences in pre- or postoperative conditions between groups, and similar surgical outcomes were achieved with the WAVS and conventional procedures. However, compared with the conventional procedure, the WAVS procedure resulted in fewer intraoperative corneal epithelial disorders (p = 0.049) and decreased the surgical duration of segmental buckling (p = 0.02); therefore, it may be suggested as an effective alternative procedure.

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Vision loss by central retinal vein occlusion after kaatsu training

Ozawa Yoko, Koto Takashi, Shinoda Hajime, Tsubota Kazuo

Medicine (United States) 94 ( 36 ) 2015.09

ISSN 0025-7974

Kaatsu training is an exercise method involving the application of pressure to the target muscle, and is being increasingly used in rehabilitation programs for heart disease patients in some hospitals. This method restricts blood flow to the muscles during exercise, and the resultant hypoxia effectively causes muscle hypertrophy and strengthening. However, no medical guidelines or risk factors for its use have been established. We report a case involving a 45-year-old man who suffered from 2 episodes of central retinal vein occlusion (CRVO), both occurring on the day following a Kaatsu training session. As a characteristic of the CRVO and its subsequent complications, the affected eye lost vision despite treatment. The patient had a history of hypertension and diabetes, and thus was at an increased risk of CRVO. Kaatsu training, which changes the heart rate and serum growth hormone levels, may have triggered the onset of CRVO. This case highlights that underlying medical conditions such as hypertension, diabetes, and the consequent inflammation, could be risk factors for vascular side effects resulting from Kaatsu training. Further studies are required before the medical and recreational use of Kaatsu training become widespread.

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Diquafosol sodium ophthalmic solution for the treatment of dry eye

Shigeyasu Chika, Yamada Masakazu, Akune Yoko, Tsubota Kazuo

Japanese Journal of Ophthalmology 2015.08

ISSN 0021-5155

Purpose: To evaluate the clinical efficacy of 3 % diquafosol sodium ophthalmic solution for dry eye, and to analyze the concentration of tear proteins and mucin-like substances after the treatment. Methods: Fifty eyes of 25 patients with dry eye syndrome were prospectively enrolled. The patients were treated with diquafosol solution at a dose of 1 drop in each eye 6 times daily for 4 weeks. The parameters of clinical efficacy were tear osmolarity, tear breakup time (BUT), fluorescein staining scores for the cornea and conjunctiva, Schirmer test values, and subjective symptoms evaluated using the ocular surface disease index (OSDI). Tears collected with Schirmer test strips were analyzed by high-performance liquid chromatography, and the concentrations of the total protein and the 4 major tear proteins, namely, secretory IgA, lactoferrin, lipocalin-1, lysozyme, and N-acetyl-neuraminic acid (Neu5Ac), were measured. Neu5Ac is a major sialic acid, a marker of secretory mucins. Results: The BUT, keratoconjunctival staining scores, and Schirmer test values were improved with statistical significance after the treatment with diquafosol solution, while changes in the other parameters, including tear osmolarity, corneal staining scores, and OSDI scores were not significant. The Neu5Ac concentration was significantly increased, which was not accompanied by changes in tear proteins. Conclusions: Topical application of diquafosol significantly improved the clinical parameters of the BUT, keratoconjunctival staining scores, and Schirmer test values and was accompanied by increased sialic acid content in the tears of patients with dry eye.

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Scleral thinning after eye-whitening surgery

Ide Takeshi, Ichihashi Yoshiyuki, Toda Ikuko, Fukumoto Teruki, Tsubota Kazuo

Asian Journal of Ophthalmology 14 ( 2 ) 68 - 72 2015.08

ISSN 1560-2133

Effectiveness of autologous serum eye drops combined with punctal plugs for the treatment of Sjögren syndrome-related dry eye

Liu Ying, Hirayama Masatoshi, Cui Xin, Connell Samuel, Kawakita Tetsuya, Tsubota Kazuo

Cornea 34 ( 10 ) 1214 - 1220 2015.07

ISSN 0277-3740

Purpose: To evaluate the efficacy and safety of autologous serum (AS) eye drops combined with punctal plugs (PPs) in patients with Sjögren syndrome (SS)-related dry eye. Methods: A retrospective clinical study was performed in patients with dry eye caused by SS. We evaluated the Schirmer test value, tear breakup time (tBUT), and fluorescein and Rose Bengal (RB) staining scores at baseline, 3 months, 6 months, 1 year, and >1 year after treatment. The dry eye indexes were also evaluated in 2 subgroups, which determined by the using of PPs, including the AS + PP group and AS only group. Results: A total of 56 eyes of 28 patients were investigated with a mean follow-up of 42.3 ± 26.1 months. After the application of AS eye drops, the Schirmer test showed no significant changes. The tBUT (2.7 ± 1.9 seconds) was significantly improved at each time point (3.9 ± 3.1, 4.5 ± 3.1, 3.7 ± 2.5, and 5.1 ± 4.0; P < 0.01), fluorescein staining (4.3 ± 2.8) was significantly improved at each time point (2.2 ± 2.2, 1.9 ± 1.9, 1.8 ± 1.6, and 1.8 ± 1.8; P < 0.01), and RB (2.6 ± 3.0) staining was significantly improved from 6-month treatment (1.5 ± 1.9, 1.9 ± 1.9, and 1.4 ± 1.8; P < 0.05, 0.01, and 0.01, respectively). When combined with PPs, the tBUT and RB staining scores were found to be significantly improved in the AS + PP group compared with those of the AS only group. Conclusions: Long-term application of AS eye drops was found to be an effective and apparently safe treatment for SS dry eye. Furthermore, PPs in combination with AS eye drops were considered to have an additive effect on SS dry eye. ©

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Analysis of Meibum before and after Intraductal Meibomian Gland Probing in Eyes with Obstructive Meibomian Gland Dysfunction

Nakayama Naohiko, Kawashima Motoko, Kaido Minako, Arita Reiko, Tsubota Kazuo

Cornea 34 ( 10 ) 1206 - 1208 2015.07

ISSN 0277-3740

Purpose: To evaluate whether the amount of meibum and its viscosity change after intraductal meibomian gland probing in patients with refractory obstructive meibomian gland dysfunction (o-MGD). Methods: Six lid margins of 3 patients with refractory o-MGD underwent intraductal meibomian gland probing. Meibum and the clinical outcome were evaluated before the procedure and at a 1-month postoperative visit. Meibum was quantified with a Meibometer, and its viscosity (Shimazaki grade) was assessed simultaneously. The tear film condition was evaluated by lipid layer interferometry (DR1, Kowa, Nagoya, Japan), and meibomian gland loss was analyzed by noncontact infrared meibography. Lid margin findings, tear break-up time, fluorescein score, and ocular symptoms were also assessed. Results: At the postoperative visits, all cases showed improvements in meibum lipid levels (446-1376, 757-802, and 396-571 meibometer units) and meibum viscosity (grade 3-0, 3-1, and 3-2). Two cases showed an improvement in tear break-up time (2-5 and 0-6 seconds). No morphological changes in the meibomian gland were observed in any cases. Conclusions: Intraductal meibomian gland probing seems to improve meibomian gland lipid levels, and it may be a good treatment option for cases of o-MGD that are resistant to conventional treatment.

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Development of a transgenic mouse with R124H human TGFBI mutation associated with granular corneal dystrophy type 2

Yamazoe Katsuya, Yoshida Satoru, Yasuda Miyuki, Hatou Shin, Inagaki Emi, Ogawa Yoko, Tsubota Kazuo, Shimmura Shigeto

PLoS ONE 10 ( 7 ) 2015.07

Purpose To investigate the phenotype and predisposing factors of a granular corneal dystrophy type 2 transgenic mouse model. Methods Human TGFBI cDNA with R124H mutation was used to make a transgenic mouse expressing human protein (TGFBI<inf>R124H</inf> mouse). Reverse transcription PCR (RT-PCR) was performed to analyze TGFBI<inf>R124H</inf> expression. A total of 226 mice including 23 homozygotes, 106 heterozygotes and 97 wild-type mice were examined for phenotype. Affected mice were also examined by histology, immunohistochemistry and electron microcopy. Results RT-PCR confirmed the expression of TGFBI<inf>R124H</inf> in transgenic mice. Corneal opacity defined as granular and lattice deposits was observed in 45.0% of homozygotes, 19.4% of heterozygotes. The incidence of corneal opacity was significantly higher in homozygotes than in heterozygotes (p = 0.02). Histology of affected mice was similar to histology of human disease. Lesions were Congo red and Masson Trichrome positive, and were observed as a deposit of amorphous material by electron microscopy. Subepithelial stroma was also stained with thioflavin T and LC3, a marker of autophagy activation. The incidence of corneal opacity was higher in aged mice in each group. Homozygotes were not necessarily more severe than heterozygotes, which deffers from human cases. Conclusions We established a granular corneal dystrophy type 2 mouse model caused by R124H mutation of human TGFBI. Although the phenotype of this mouse model is not equivalent to that in humans, further studies using this model may help elucidate the pathophysiology of this disease.

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Investigating the influence of visual function and systemic risk factors on falls and injurious falls in glaucoma using the structural equation modeling

Yuki Kenya, Asaoka Ryo, Tsubota Kazuo

PLoS ONE 10 ( 6 ) 2015.06

Purpose: To investigate the relationship between visual function and the risks of falling and injurious falls in subjects with primary open angle glaucoma (POAG) Methods: Questionnaires were conducted in 365 POAG patients to assess history of falls and falls with injury and general patient health. Structural equation modeling (SEM) was used to investigate the relationship between visual function, as measured by a patient's binocular integrated visual field and visual acuity (VA), general health and the risks of falling and injurious falls. Results: Among the 365 subjects, 55 subjects experienced falls in the past year. A significant difference was observed in worse-eye VA between the faller and non-faller groups (p = 0.03). SEM of fallers obtained a Root Mean Square Error of Approximation (RMSEA) of 0.035 and a Comparative Fit Index (CFI) of 0.99. The 95% confidence intervals (CI) of regression coefficients from this model suggested better VA and worse VA were significant risk factors for falling. Among the 55 fallers, 22 subjects experienced an associated injury. There was a significant difference in gender between the non-injurious and injurious faller groups (p = 0.002). SEM of injurious fallers obtained a RMSEA of 0.074 and a CFI of 0.97. In this SEM model, the 95% CI of regression coefficients suggested gender and average total deviation values in the lower peripheral visual field were significant risk factors for an injurious fall. Conclusions: This study suggests that worse-eye and better-eye VAs are associated with falls. Furthermore, patients with inferior visual field loss and females were found to be at greater risk of injurious falls.

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High prevalence of sleep and mood disorders in dry eye patients

Ayaki Masahiko, Kawashima Motoko, Negishi Kazuno, Tsubota Kazuo

Neuropsychiatric Disease and Treatment 11 889 - 894 2015.05

ISSN 1176-6328

Purpose: We aimed to explore the prevalence of probable sleep and mood disorders in eye clinic visitors. Methods: This was a cross-sectional study. The participants were outpatients at six eye clinics from January through March, 2014. Outpatients were invited to complete a questionnaire containing the Pittsburgh Sleep Quality Index (PSQI) and Hospital Anxiety and Depression Scale (HADS). A final diagnosis was made, and patients were classified into six diagnostic groups. The main outcome measures were the scores of the PSQI and HADS among the diagnostic groups. Results: A total 1,000 outpatients participated, and 730 patients (mean age 59.5±19.0 years; 487 females) were analyzed after exclusion of children and patients diagnosed with healthy eyes, acute injury, or unilateral pseudophakia. The mean PSQI and HADS scores across all patients were 5.3±3.1 and 9.2±6.2, respectively. For the diagnostic groups, the mean PSQI and HADS scores, respectively, were 5.7±3.3 and 10.2±6.0 for dry eye (n=247), 5.4±3.2 and 9.2±5.7 for bilateral cataracts (n=159), 5.3±3.3 and 8.0±5.3 for bilateral pseudophakia (n=99), and, 5.0±3.1 and 9.8±6.6 for glaucoma (n=109). Overall, 37.3% of patients were poor sleepers (PSQI $6), and 45.5% had possible mood disorders (HADS $10). Stepwise regression analysis revealed that the PSQI and HADS scores were significantly correlated with both age (P,0.05) and the presence of dry eye (P,0.05). Conclusion: The prevalence of sleep and mood disorders was significantly higher in patients with dry eye. The present results suggest consultation-liaison psychiatry services may be beneficial among eye disease patients.

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Correction

Uchino Yuichi, Kawakita Tetsuya, Miyazawa Masaki, Ishii Takamasa, Onouchi Hiromi, Yasuda Kayo, Ogawa Yoko, Shimmura Shigeto, Ishii Naoaki, Tsubota Kazuo

PLoS ONE 10 ( 5 ) 2015.05

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Increased tear fluid production as a compensatory response to meibomian gland loss

Arita Reiko, Morishige Naoyuki, Koh Shizuka, Shirakawa Rika, Kawashima Motoko, Sakimoto Tohru, Suzuki Takashi, Tsubota Kazuo

Ophthalmology 122 ( 5 ) 925 - 933 2015.05

ISSN 0161-6420

Purpose To compare tear film parameters as well as meibomian gland morphologic features and function among patients with meibomian gland dysfunction (MGD), those with non-Sjögren syndrome aqueous-deficient dry eye (non-SS ADDE), those with non-SS ADDE and MGD, and normal subjects. Design Multicenter, cross-sectional, observational case series. Participants Forty-one eyes of 41 patients (all women; mean age ± standard deviation, 62.1±9.9 years) with non-SS ADDE, 70 eyes of 70 patients (all women; 66.0±8.7 years) with MGD, 17 eyes of 17 patients (all women; 72.4±7.8 years) with non-SS ADDE and MGD, and 70 eyes of 70 normal control subjects (all women; 65.0±7.1 years). Methods Ocular symptoms were scored from 0 to 14 and lid margin abnormalities from 0 to 4 according to their respective number. Meibomian gland changes were scored from 0 to 6 (meiboscore) on the basis of noncontact meibography findings, and meibum was graded from 0 to 3 depending on its volume and quality. Conjunctival and corneal epithelial damage were scored from 0 to 9 (fluorescein score). Tear film break-up time (TBUT) was measured as an index of tear film stability, and tear fluid production was evaluated with Schirmer's test. Main Outcome Measures Ocular symptom score, lid margin abnormality score, meiboscore, meibum grade, fluorescein score, TBUT, and Schirmer's test value. Results The ocular symptom score did not differ significantly between the MGD and non-SS ADDE groups (P = 0.762). The lid margin abnormality score, meiboscore, and meibum grade were significantly higher in the MGD group than in the non-SS ADDE group (P = 0.0012, P < 0.0001, and P < 0.0001, respectively). The fluorescein score, TBUT, and Schirmer's test value were significantly worse in the non-SS ADDE group than in the MGD group (P < 0.0001, P = 0.0061, and P < 0.0001, respectively). The meiboscore correlated significantly with Schirmer's test value only in the MGD group (ρ = 0.508, P = 8.3×10-6). Conclusions An increase in tear fluid production likely compensates for loss of meibomian glands in individuals with MGD.

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Bilberry extract supplementation for preventing eye fatigue in video display terminal workers

Ozawa Y., Kawashima M., Inoue S., Inagaki E., Suzuki A., Ooe E., Kobayashi S., Tsubota Kazuo

Journal of Nutrition, Health and Aging 19 ( 5 ) 548 - 554 2015.05

ISSN 1279-7707

Objectives: To examine the effect of a dietary supplement containing bilberry extract (BE) on eye fatigue induced by acute video display terminal (VDT) loads.Design and setting: A prospective, randomized, double-blind, placebo-controlled study was performed from August 2012 to February 2013 in the Medical Corporation Jico-kai Yagi Hospital, and the Shinyokohama Shinoharaguchi Orthopedic Surgery and Dermatology Clinic, in Japan.Participants: Two hundred eighty-one office workers aged 20–40 years that used VDTs were screened by critical flicker fusion (CFF) and near point accommodation (NPA).Intervention: The participants were randomized to either a BE (480 mg/day) or placebo (vehicle) group, and took allocated capsule, daily, for 8 weeks.Measurements: The CFF, NPA, contrast visual acuity, functional visual acuity, keratoconjunctival epithelial damage, and fluorescein tear film break-up time were examined, and 18 subjective symptoms of eye fatigue were evaluated by questionnaire. Adverse events were reported via medical interviews. Data were collected both before and after VDT load at baseline, and 4, and 8 weeks after daily supplementation with either BE or placebo.Results: Of 281 participants screened, 88 having relatively lower levels of CFF and NPA were enrolled in the study. Of these, 37 control and 43 BE group subjects completed the study. The VDT load-induced reduction in CFF was alleviated after 8 weeks of BE supplementation (95% confidence interval, 0.10–1.60; p=0.023), in contrast to placebo supplementation, while NPA variation was not. Of the subjective symptoms of eye fatigue, VDT load-induced ocular fatigue sensation, ocular pain, eye heaviness, uncomfortable sensation, and foreign body sensation were mitigated more in the BE group than in the control group, at week 8 (p<0.05). There were no severe adverse events in either group.Conclusions: BE supplementation improved some of the objective and subjective parameters of eye fatigue induced by VDT loads.

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Advanced dry eye screening for visual display terminal workers using functional visual acuity measurement

Kaido Minako, Kawashima Motoko, Yokoi Norihiko, Fukui Masaki, Ichihashi Yoshiyuki, Kato Hiroaki, Yamatsuji Motoko, Nishida Mitsuko, Fukagawa Kazumi, Kinoshita Shigeru, Tsubota Kazuo

British Journal of Ophthalmology 2015.04

ISSN 0007-1161

Purpose To evaluate the validity of a dry eye (DE) screening method in visual display terminal (VDT) workers using functional visual acuity (VA) measurement combined with a symptoms questionnaire, and to make recommendations regarding practical use of the screening. Methods This prospective cross-sectional study included 369 young and middle-age office workers, who use VDTs. Subjects completed two types of DE questionnaires and underwent DE testing and functional VA measurement. Subjects were assigned to DE and non-DE groups. Sensitivity and specificity of the DE prediction value were assessed. According to the probability distribution of DE and non-DE calculated by the discriminant analysis formula, we classified subjects into three categories based on the most appropriate cut-off levels. Positive predictive value (PPV) and negative predictive value (NPV) of the DE prediction value were also assessed. Results Of 369 subjects, 218 and 151 subjects were assigned to the DE and non-DE groups, respectively. Sensitivity and specificity were 93.1% and 43.7%, respectively. Based on the probability prediction, the cut-off levels were defined as probability prediction ≥85%, 45%≤ probability prediction <85% and probability prediction <45%. Subjects with a probability prediction ≥85% were defined positive (N=130), while those with values <45% were defined as negative (N=73). The PPV and NPV were 83.8% and 80.8%, respectively. Conclusions Acceptable PPV and NPV were obtained by setting three discriminate categories. This improved version of the DE screening method may be acceptable for practical use in VDT workers.

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Novel corneal piggyback technique for consecutive intraocular lens implantation and penetrating keratoplasty surgery

Matsumoto Yukihiro, Dogru Murat, Shimazaki Jun, Tsubota Kazuo

Cornea 34 ( 6 ) 713 - 716 2015.04

ISSN 0277-3740

Purpose: To report a novel "piggyback" penetrating keratoplasty technique performed in a patient with severe keratoconus. Methods: A 43-year-old man underwent cataract surgery and a new "piggyback" penetrating keratoplasty (PKP) technique in his right eye, as he suffered from severe keratoconus and mature cataract due to severe atopic dermatitis. Under general anesthesia, phacoemulsification and aspiration (PEA), intraocular lens (IOL) implantation, and "piggyback" PKP while avoiding open-sky surgery was performed to prevent serious complications including IOL and vitreous prolapse. This PKP technique had been completed as a totally closed surgery. Results: One year after "piggyback" PKP and cataract surgery, the grafted cornea remained clear and IOL was also well positioned. Best-corrected visual acuity improved from hand motions to 30/100. Conclusions: A novel "piggyback" PKP technique was successfully performed in a patient with severe keratoconus. The new "piggyback" corneal transplantation technique may become an indispensable tool for transplant surgeons who want to improve surgical safety and predictability.

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Importance of tear film instability in dry eye disease in office workers using visual display terminals

Yokoi Norihiko, Uchino Miki, Uchino Yuichi, Dogru Murat, Kawashima Motoko, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Tsubota Kazuo, Kinoshita Shigeru

American Journal of Ophthalmology 159 ( 4 ) 748 - 754 2015.04

ISSN 0002-9394

Purpose To evaluate the relationship between subjective symptoms and clinical signs in dry eye disease (DED) in office workers using visual display terminals (VDTs). Design Cross-sectional study. Methods This study involved 672 Japanese young and middle-aged office workers who use VDTs. The subjects completed questionnaires designed to detect subjective symptoms and risk factors for DED. Dry eye tests, including tear film break-up time (TBUT), corneal-conjunctival staining with fluorescein and lissamine green, and the Schirmer test, were performed. Based on the Japanese diagnostic criteria for DED, the subjects were classified into 3 groups: definite DED, probable DED, and non-DED. Between each group, subjective symptoms and clinical signs were compared. Results Of the 672 subjects, 561 (374 male, 187 female) completed the questionnaire (response rate: 83.5%). Definite DED was diagnosed in 65 subjects (11.6%), probable DED in 303 subjects (54.0%), and non-DED in 193 subjects (34.4%). The mean subjective symptom score was significantly less in subjects with probable DED (2.05 ± 0.42) and non-DED (1.63 ± 0.38) than in those with definite DED (2.19 ± 0.40) (P <.05 and P <.01, respectively). In the subjects with probable DED, a subgroup with positive subjective symptoms and abnormal TBUT (≤5 seconds) was categorized as short TBUT-type DED, and it was found that they had a higher subjective symptom score (2.09 ± 0.40), equivalent to that of those with definite DED (P =.269). Conclusions Despite no or minor epithelial damage, the severity of subjective symptoms was greater in short TBUT-type DED, most likely attributable to tear film instability. Thus, it might prove important to evaluate TBUT to successfully treat those patients.

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Color of Intra-Ocular Lens and Cataract Type Are Prognostic Determinants of Health Indices After Visual and Photoreceptive Restoration by Surgery

Ayaki Masahiko, Negishi Kazuno, Suzukamo Yoshimi, Tsubota Kazuo

Rejuvenation Research 18 ( 2 ) 145 - 152 2015.04

ISSN 1549-1684

Background: This study compared post-operative quality of life and sleep according to the type of cataract opacity and color of the implanted intra-ocular lens (IOL). Methods: This is a cohort study and participants were 206 patients (average age 74.1 years) undergoing cataract surgery with the implantation of a clear ultra-violet (UV)-blocking IOL (C) or a yellow blue-light-blocking IOL (Y). Participants were evaluated using the National Eye Institute Visual Function Questionnaire (VFQ-25) and Pittsburgh Sleep Quality Index (PSQI) before surgery and 2 and 7 months after surgery. Changes in sub-scale scores of VFQ-25 and PSQI were compared. Results: Sub-scale analyses for improvement after surgery revealed significant differences in ocular pain scores on the VFQ-25 (Y>C; the higher the score, the better the outcome). Furthermore, there were significant differences between the two IOLs in terms of the sleep latency score (C>Y) and sleep disturbances score (C>Y). A posterior sub-capsular cataract was significantly correlated with improvements in ocular pain and sleep latency scores. These effects were successfully represented by the change in scores rather than absolute post-operative scores because individual standard of response may often change after intervention, recognized as a response shift phenomenon in patient-reported outcome study. Regarding seasonal differences, patients who had surgery in summer exhibited relatively better sleep quality than those who had surgery in winter. Conclusions: Analysis of sub-scales of health indices demonstrated characteristic prognoses for each IOL and cataract type. Cataract surgery may potentially contribute to systemic health in older adults.

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Associations between subjective happiness and dry eye disease

Kawashima Motoko, Uchino Miki, Yokoi Norihiko, Uchino Yuichi, Dogru Murat, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Kinoshita Shigeru, Mimura Masaru, Tsubota Kazuo

PLoS ONE 10 ( 4 ) 2015.04

Importance: Dry eye disease has become an important health problem. A lack of concordance between self-reported symptoms and the outcome of dry eye examinations has raised questions about dry eye disease. Objective: To explore the association between subjective happiness and objective and subjective symptoms of dry eye disease. Design: The study adopted a cross-sectional design. Setting: All the employees of a company in Osaka, Japan. Participants: 672 Japanese office workers using Visual Display Terminals (age range: 26-64 years). Methods: The dry eye measurement tools included the Schirmer test, conjunctivocorneal staining, the tear film break-up time, as well as the administration of a dry eye symptoms questionnaire. Happiness was measured by the Subjective Happiness Scale. Main Outcome Measures: Dry eye examination parameters, dry eye symptoms questionnaires, and the Subjective Happiness Scale score. Results: Of the 672 workers, 561 (83.5%) completed the questionnaires and examinations. The mean Subjective Happiness Scale score was 4.91 (SD = 1.01). This score was inversely correlated with the dry eye symptom score (r = -0.188, p < 0.001), but was not associated with objective findings which include conjunctivocorneal staining, low Schirmer test score, or low tear film break-up time. The level of subjective happiness was the lowest in the group without objective results, but reported subjective symptoms of dry eyes (p < 0.05). Conclusions and Relevance: There is evidence of the relationship between subjective happiness and self-reported symptoms of dry eyes. Findings of this study revealed a new perspective on dry eye disease, including the potential for innovative treatments of a specific population with dry eye disease.

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Predisposing factors for intraocular lens dislocation treated by pars plana vitrectomy

Yotsukura-Tsutsui Erisa, Koto Takashi, Tsubota Kazuo, Ozawa Yoko

Journal of Cataract and Refractive Surgery 41 ( 4 ) 892 - 894 2015.04

ISSN 0886-3350

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Association of macular pigment optical density with serum concentration of oxidized low-density lipoprotein in healthy adults

Nagai Norihiro, Izumi-Nagai Kanako, Suzuki Misa, Shinoda Hajime, Koto Takashi, Uchida Atsuro, Mochimaru Hiroshi, Tomita Yohei, Miyake Seiji, Kobayashi Saori, Sasaki Mariko, Tsubota Kazuo, Ozawa Yoko

Retina 35 ( 4 ) 820 - 826 2015.04

ISSN 0275-004X

Purpose: To analyze the association between macular pigment optical density (MPOD), which reflects lutein (L), zeaxanthin (Z), and meso-zeaxanthin (MZ) in the macula, and background characteristics. Methods: Fifty-five healthy adult volunteers were analyzed. Macular pigment optical density was measured using a heterochromatic flicker photometry technique, and serum concentrations of carotenoids and lipoproteins were by high-performance liquid chromatography and enzyme-linked immunosorbent assay, respectively. Dietary intake of nutrient was determined by a validated self-administered questionnaire on ingestion frequency. Results: Macular pigment optical density was positively correlated with serum concentrations of L and Z and dietary L intake and inversely correlated with serum oxidized low-density lipoprotein (LDL). Although MPOD decreased with age (95% confidence interval, -0.011 to -0.002; correlation coefficient, -0.269; P 0.007), serum L/Z and dietary L intake did not. In contrast, serum oxidized LDL was positively correlated with age (95% confidence interval, 0.69-2.34; correlation coefficient, 0.333; P 0.0004). After adjusting for age, sex, and oxidized LDL, serum L was positively correlated with MPOD (95% confidence interval, 0.88-1.69; P 0.000001). After adjusting for age, sex, and serum L, serum oxidized LDL was inversely correlated with MPOD (95% confidence interval, -0.002 to -0.0004; P 0.006). Conclusion: Macular pigment optical density was inversely correlated with serum oxidized LDL. Further study to know the impact of oxidized LDL on MPOD may be warranted.

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Driving self-restriction and motor vehicle collision occurrence in glaucoma

Ono Takeshi, Yuki Kenya, Awano-Tanabe Sachiko, Fukagawa Kazumi, Shimoyama Masaru, Ozawa Yoko, Ozeki Naoki, Shiba Daisuke, Tsubota Kazuo

Optometry and Vision Science 92 ( 3 ) 357 - 364 2015.03

ISSN 1040-5488

Purpose. To confirm that subjects with primary open-angle glaucoma (POAG) who avoid driving in high-risk situations are less likely to be involved in motor vehicle collisions (MVCs) than those who do not. Methods. This study evaluated 252 consecutive Japanese aged between 40 and 85 years with POAG. All participants were requested to answer a questionnaire on their driving habits, including self-restriction in driving at night, in rain, in fog, on freeways, and lane changing, and history of MVCs. Those who reported restricting their driving in one or more ways constituted the self-restriction group, and those who reported no self-restriction made up the no-restriction group. The prevalence of MVCs and the crash rate (number of MVCs/10,000 km driven) were compared between the two groups. The association between prevalence of MVCs and the number of driving self-restrictions was also evaluated. Results. The association between driving self-restriction and MVCs was observed among the male subjects, not among the female subjects. Among the male subjects, the prevalence of MVCs was significantly higher in the no-restriction group than in the self-restriction group (no-restriction group, 33/107 = 30.8%; self-restriction group, 9/66 = 13.6%, p = 0.01). The crash rate was also significantly higher in the no-restriction group (no-restriction group, 1.4 T 0.8; self-restriction group, 0.4 T 0.3, average T SE, p = 0.01). No restriction was significantly associated with MVCs (multivariable-adjusted odds ratios, 2.43 [95% confidence interval, 1.03 to 5.73]). The number of driving self-restrictions was also associated with MVCs (multivariable- adjusted odds ratios, 0.41 [95% confidence interval, 0.18 to 0.99], per one increment of self-restriction). Conclusions. Driving self-restriction may be associated with a reduced prevalence of MVCs in men with POAG.

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The effect of Nrf2 knockout on ocular surface protection from acute tobacco smoke exposure

Kojima Takashi, Dogru Murat, Higuchi Akihiro, Nagata Taeko, Ibrahim Osama M A, Inaba Takaaki, Tsubota Kazuo

American Journal of Pathology 185 ( 3 ) 776 - 785 2015.03

ISSN 0002-9440

Ocular surface mucosa is the first-line ocular tissue to be exposed to environmental stress. We evaluated tear functions and keratoconjunctival epithelial alterations after sidestream cigarette smoke (SCS) exposure and tried to clarify the role of the transcription factor nuclear factor erythroid 2-related factor 2 (Nfe2l2, also known as Nrf2), on the ocular surface. In wild-type and Nrf2^-/- mice, tear volume did not change after SCS exposure. Tear film breakup time (tear stability) in Nrf2^-/- mice was significantly shorter than that in wild-type mice after SCS exposure. Vital staining scores, including fluorescein and Rose Bengal staining, showed significantly higher values in Nrf2^-/- mice than in wild-type mice after SCS exposure. Excessive oxidative stress accumulation was detected in Nrf2^-/- mice after SCS exposure using immunohistochemical analysis. Immunohistochemical analysis also revealed decreased mucin 1 (Muc1) and Muc5ac staining in Nrf2^-/- mice after SCS exposure. mRNA expression levels of Muc1, Muc4, and Muc5ac and of SAM-pointed domain epithelial-specific transcription factor in Nrf2^-/- mice were lower than those in wild-type mice after SCS exposure. Mean tear IL-6 concentrations increased significantly in Nrf2^-/- mice after SCS exposure. In conclusion, SCS exposure induced decreased tear stability, ocular surface damage, and altered conjunctival phenotype in Nrf2^-/- mice. Nrf2 could play an important role in protection of the ocular surface against SCS exposure.

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Blue light-induced inflammatory marker expression in the retinal pigment epithelium-choroid of mice and the protective effect of a yellow intraocular lens material invivo

Narimatsu Toshio, Negishi Kazuno, Miyake Seiji, Hirasawa Manabu, Osada Hideto, Kurihara Toshihide, Tsubota Kazuo, Ozawa Yoko

Experimental Eye Research 132 48 - 51 2015.03

ISSN 0014-4835

Oxidative stress in the retinal pigment epithelium (RPE) is a well-accepted pathogenic change in vision-threatening diseases such as age-related macular degeneration. One source of oxidative stress is excessive light exposure, which causes excessive activation of the visual cycle. Because short wavelength light (blue light) has more energy, it is reported to be more harmful to photoreceptor cells than the other wavelengths of light. However, the biological effect of blue light in the RPE of living animals and the protective effect of a yellow intraocular lens (IOL) material that blocks blue light is still obscure. Therefore, we compared the pathogenic effect in the RPE-choroid complexes of mice exposed to light in a box made of a clear or a yellow IOL material. We measured the level of reactive oxygen species (ROS) using 2', 7'-dichlorodihydrofluorescein diacetate, the mRNA levels of inflammatory cytokines and a macrophage marker by real-time polymerase chain reaction, and the protein level of monocyte chemotactic protein-1 (MCP-1) by ELISA. The ROS level after light exposure was suppressed in the RPE-choroids of light-exposed mice in the yellow IOL material box. In parallel, all the inflammatory cytokines that we measured and a macrophage marker were also suppressed in the RPE-choroids of light-exposed mice in the yellow IOL material box. Therefore, a yellow IOL material suppressed, and thus blue light exacerbated, the increase in the ROS level and inflammatory cytokine expression as well as macrophage recruitment in the RPE-choroid invivo after light exposure.

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Wickerhamomyces anomalus fungal keratitis responds to topical treatment with antifungal micafungin

Kamoshita Mamoru, Matsumoto Yukihiro, Nishimura Kazuko, Katono Yasuhiro, Murata Mitsuru, Ozawa Yoko, Shimmura Shigeto, Tsubota Kazuo

Journal of Infection and Chemotherapy 21 ( 2 ) 141 - 143 2015.02

ISSN 1341-321X

We describe a 91-year-old woman who suffered from fungal keratitis after corneal transplantation. The causative organism was identified as Wickerhamomyces anomalus (formerly Pichia anomala or Hansenula anomala) on the basis of morphological characteristics and the sequence of the internal transcribed spacer region of the ribosomal RNA gene. The patient was successfully treated with topical micafungin (MCFG) only. We present the first report of a case of W.anomalus fungal keratitis that responded to topical treatment with the antifungal MCFG.

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Clinical and molecular characteristics of childhood-onset stargardt disease

Fujinami Kaoru, Zernant Jana, Chana Ravinder K., Wright Genevieve A., Tsunoda Kazushige, Ozawa Yoko, Tsubota Kazuo, Robson Anthony G., Holder Graham E., Allikmets Rando, Michaelides Michel, Moore Anthony T.

Ophthalmology 122 ( 2 ) 326 - 334 2015.02

ISSN 0161-6420

Purpose To describe the clinical and molecular characteristics of patients with childhood-onset Stargardt disease (STGD). Design Retrospective case series. Participants Forty-two patients who were diagnosed with STGD in childhood at a single institution between January 2001 and January 2012. Methods A detailed history and a comprehensive ophthalmic examination were undertaken, including color fundus photography, autofluorescence imaging, spectral-domain optical coherence tomography (SD-OCT), and pattern and full-field electroretinograms. The entire coding region and splice sites of ABCA4 were screened using a next-generation, sequencing-based strategy. The molecular genetic findings of childhood-onset STGD patients were compared with those of adult-onset patients. Main Outcome Measures Clinical, imaging, electrophysiologic, and molecular genetic findings. Results The median ages of onset and the median age at baseline examination were 8.5 (range, 3-16) and 12.0 years (range, 7-16), respectively. The median baseline logarithm of the minimum angle of resolution visual acuity was 0.74. At baseline, 26 of 39 patients (67%) with available photographs had macular atrophy with macular/peripheral flecks; 11 (28%) had macular atrophy without flecks; 1 (2.5%) had numerous flecks without macular atrophy; and 1 (2.5%) had a normal fundus appearance. Flecks were not identified at baseline in 12 patients (31%). SD-OCT detected foveal outer retinal disruption in all 21 patients with available images. Electrophysiologic assessment demonstrated retinal dysfunction confined to the macula in 9 patients (36%), macular and generalized cone dysfunction in 1 subject (4%), and macular and generalized cone and rod dysfunction in 15 individuals (60%). At least 1 disease-causing ABCA4 variant was identified in 38 patients (90%), including 13 novel variants; ≥2 variants were identified in 34 patients (81%). Patients with childhood-onset STGD more frequently harbored 2 deleterious variants (18% vs 5%) compared with patients with adult-onset STGD. Conclusions Childhood-onset STGD is associated with severe visual loss, early morphologic changes, and often generalized retinal dysfunction, despite often having less severe fundus abnormalities on examination. One third of children do not have flecks at presentation. The relatively high proportion of deleterious ABCA4 variants supports the hypothesis that earlier onset disease is often owing to more severe variants in ABCA4 than those found in adult-onset disease.

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Fluorophotometric Analysis of the Ocular Surface Glycocalyx in Soft Contact Lens Wearers

Fukui Masaki, Yamada Masakazu, Akune Yoko, Shigeyasu Chika, Tsubota Kazuo

Current Eye Research 2015.01

ISSN 0271-3683

Purpose: Unstable tear film characterized by shorting of tear break-up time (BUT) is associated with discomfort and dryness in contact lens wearers. The glycocalyx is thought to be crucial in maintaining the wettability and lubrication of the ocular surface. We evaluated the ocular surface glycocalyx in soft contact lens (SCL) wearers using a fluorescein-labeled wheat germ agglutinin (F-WGA) as a marker to demonstrate the ocular surface glycoconjugates in vivo. Methods: Twenty experienced SCL wearers and 20 healthy volunteers with no history of CL wearing (controls) were enrolled in the study. After applying a 5% F-WGA solution to the eyes of study individuals, fluorescent intensities in their respective central corneas were measured by fluorophotometry. The relationship between F-WGA intensity in the corneal surface and clinical parameters associated with contact lens wear were analyzed. Results: F-WGA fluorescence intensity in the SCL group was 418.5 ± 103.3, which was significantly lower than that of the controls (825.0 ± 179.8; p < 0.0001, Mann–Whitney test). F-WGA fluorescence intensity was not correlated with Schirmer’s test values or age, whereas a statistically significant correlation between F-WGA fluorescence intensity and tear film BUT was observed (r = 0.77, p < 0.0001). The decrease in F-WGA fluorescence intensity could be reversed by discontinuation of SCL use. Conclusion: Reduction and/or compositional alteration of ocular surface glycocalyx may be one of the causative factors of SCL-induced eye dryness.

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Abdominal breathing increases tear secretion in healthy women

Sano Kokoro, Kawashima Motoko, Ikeura Kazuhiro, Arita Reiko, Tsubota Kazuo

Ocular Surface 13 ( 1 ) 82 - 87 2015.01

ISSN 1542-0124

Purpose To determine the relationship between abdominal breathing and tear meniscus volume in healthy women, we investigated the change in tear meniscus volume in two groups: normal breathing and abdominal breathing. Methods We used a crossover experimental model and examined 20 healthy women aged 20-54 years (mean ± SD, 32.7 ± 11.1 years). The participants were randomly assigned to one of two groups. During the first visit, the normal breathing group was subjected to normal breathing for 3 min, whereas the abdominal breathing group was subjected to abdominal breathing (4-second inhalation and 6-second exhalation) for 3 min. During the second visit, the protocols were swapped between the two groups. We estimated the R wave to R wave (R-R) interval, tear meniscus volume, salivary amylase activity, pulse, and blood pressure before and immediately after, 15 min after, and 30 min after completion of the breathing activity. Results After abdominal breathing, compared to that before breathing, the tear meniscus volume increased significantly 15 min after breathing (P<.01). Furthermore, systolic blood pressure showed a significant decrease immediately after abdominal breathing (P<0.05). No significant difference was found in the test parameters in the normal breathing group. Conclusion Abdominal breathing for 3 minutes increases the tear meniscus volume in healthy women. Consequently, abdominal breathing may be considered in the treatment of dry eye disease.

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Screening of dry eye disease in visual display terminal workers during occupational health examinations

Kawashima Motoko, Yamatsuji Motoko, Yokoi Norihiko, Fukui Masaki, Ichihashi Yoshiyuki, Kato Hiroaki, Nishida Mitsuko, Uchino Miki, Kinoshita Shigeru, Tsubota Kazuo

Journal of Occupational Health 57 ( 3 ) 253 - 258 2015

ISSN 1341-9145

Screening of dry eye disease in visual display terminal workers during occupational health examinations: The Moriguchi study: Motoko KAWASHIMA, et al. Department of Ophthalmology, Keio University School of Medicine-Purpose: The aim of this study was to estimate the prevalence of dry eye disease (DED) in Japanese visual display terminal (VDT) workers and identify simple methods for screening DED during occupational health examinations. Methods: This study involved 369 (331 men and 38 women; mean age, 44.4 [8.8] years) workers engaged in PC development with approximately 6 hours of VDT use daily. The participants completed questionnaires on dry eye and physical symptoms affecting QoL, and their ability to keep their eyes open for 10 seconds was tested for DED screening. They also underwent assessment of conjunctival and corneal vital staining with lissamine green and fluorescein, measurement of tear film breakup time, and Schirmer's test for DED diagnosis. Sensitivity, specificity, and positive predictive value of the screening methods were assessed. Results: The DED group included 218 (definite, 14; probable, 204) participants. They had markedly high frequencies of 11 dry eye symptoms, especially ocular fatigue (OR, 17.1; 95% CI, 9.9 to 29.7) and dry sensation (OR, 8.2; 95% CI, 3.6 to 18.4), and were more often unable to keep their eyes open for 10 s. With either ocular fatigue or dry sensation as the cutoff criterion, the sensitivity, specificity, and positive predictive value were 89.9, 66.9, and 79.7%, respectively. Conclusions: Approximately 60% of the VDT workers were diagnosed with DED on the basis of diverse eye symptoms affecting their ability to work. The dry eye symptom questionnaire and 10-s eye-opening workerstest are simple, noninvasive, and inexpensive methods for screening DED during occupational health examinations.

Glaucomatous visual field defect severity and the prevalence of motor vehicle collisions in Japanese

Ono Takeshi, Yuki Kenya, Asaoka Ryo, Kouyama Keisuke, Abe Takayuki, Tanabe Sachiko, Fukagawa Kazumi, Uchino Miki, Shimoyama Masaru, Ozawa Yoko, Ozeki Naoki, Shiba Daisuke, Tsubota Kazuo

Journal of Ophthalmology 2015 2015

ISSN 2090-004X

Purpose. This study examined the association between the severity of visual field defects and the prevalence of motor vehicle collisions (MVCs) in subjects with primary open-angle glaucoma (POAG). Methods. This is a cross-sectional study. Japanese patients who have had driver's licence between 40 and 85 years of age were screened for eligibility. Participants answered a questionnaire about MVCs experienced during the previous 5 years. Subjects with POAG were classified as having mild, moderate, or severe visual field defect. We evaluated associations between the severity of POAG and the prevalence of MVCs by logistic regression models. Results. The prevalence of MVCs was significantly associated with the severity of POAG categorized by worse eye MD (control: 30/187 = 16.0%; mild POAG: 17/92 = 18.5%; moderate POAG: 14/60 = 23.3%; severe POAG: 14/47 = 29.8%; P = 0.025, Cochran-Armitage trend test). Compared to the control group, the adjusted OR for MVC prevalence in subjects with mild, moderate, or severe POAG in the worse eye was 1.07 (95% CI: 0.55 to 2.10), 1.44 (95% CI: 0.68 to 3.08), and 2.28 (95% CI: 1.07 to 4.88). Conclusions. There is a significant association between the severity of glaucoma in the worse eye MD and the prevalence of MVCs.

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New eye cleansing product improves makeup-related ocular problems

Okura Masako, Kawashima Motoko, Katagiri Mikiyuki, Shirasawa Takuji, Tsubota Kazuo

Journal of Ophthalmology 2015 2015

ISSN 2090-004X

Purpose. This study evaluated the effects of using a newly developed eye cleansing formulation (Eye Shampoo) to cleanse the eyelids for 4 weeks in a parallel-group comparative study in women with chronic eye discomfort caused by heavy use of eye makeup and poor eye hygiene habits. Methods. Twenty women participants who met the inclusion criteria were randomly allocated to 2 groups comprising 10 participants each. The participants were asked to use either artificial tears alone or artificial tears in conjunction with Eye Shampoo for 4 weeks. The participants answered the questionnaire again and were reexamined, and changes in symptoms within each group and variations of symptoms between the two groups were statistically analyzed. Results. In the group using only artificial tears, improvements in subjective symptoms but not in ophthalmologic examination results were found. In the group using Eye Shampoo together with artificial tears, significant improvements were observed in the subjective symptoms, meibomian orifice obstruction, meibum secretion, tear breakup time, and superficial punctate keratopathy. Conclusion. In patients with chronic eye discomfort thought to be caused by heavy eye makeup, maintaining eyelid hygiene using Eye Shampoo caused a marked improvement in meibomian gland blockage and dry eye symptoms.

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The relationship between central visual field damage and motor vehicle collisions in primary open-angle glaucoma patients

Yuki Kenya, Asaoka Ryo, Tsubota Kazuo

PLoS ONE 9 ( 12 ) 2014.12

Purpose: To investigate the relationship between visual field (VF) damage and history of motor vehicle collisions (MVCs) in subjects with primary open-angle glaucoma (POAG). Methods: MVC history and driving habits were recorded using patient questionnaires in 247 POAG patients. Patients' driving attitudes (carefulness) were estimated using Rasch analysis. The relationship between MVC outcomes and 52 total deviation (TD) values of integrated binocular VF (IVF), better and worse visual acuities (VAs), age and gender was analyzed using principal component analysis and logistic regression. Results: 51 patients had the history of MVCs. Significant difference was observed between patients with and without history of MVCs only for: better VA, a single TD value in the superior-right VF, and the typical distance driven in a week (unpaired t-test, p=0.002, 0.015 and 0.006, respectively). There was not a significant relationship between MVCs and mean deviation (MD) of IVF (p=0.41, logistic regression). None of the principal components were significantly correlated with MVC outcome (p>0.05, polynomial logistic regression analysis). There was a significant relationship between IVF MD and Rasch derived Person parameter (R²=0.023, p=0.0095). There was also a significant positive relationship between MVCs and the distance driven in a week (p=0.005, logistic regression). Conclusions: In this study of POAG patients, MVCs were not related to central binocular VF damage. These results suggest the relationship between visual function and driving is not straightforward, and careful consideration should be given when predicting patients' driving ability using their VF.

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Resveratrol prevents the development of choroidal neovascularization by modulating AMP-activated protein kinase in macrophages and other cell types

Nagai Norihiro, Kubota Shunsuke, Tsubota Kazuo, Ozawa Yoko

Journal of Nutritional Biochemistry 25 ( 11 ) 1218 - 1225 2014.11

ISSN 0955-2863

The development of choroidal neovascularization (CNV) is a critical step in the pathogenesis of age-related macular degeneration (AMD), a vision-threatening disease. In this study, we used a mouse model of AMD to study the protective effects of resveratrol (RSV) supplementation against CNV as well as the underlying molecular mechanisms. Mice were orally pretreated with RSV daily for 5. days. On the fifth day, the mice underwent laser photocoagulation to induce CNV. One week after laser treatment, CNV volume was significantly lower in the RSV-treated mice compared with vehicle-treated animals. In addition, RSV treatment significantly inhibited macrophage infiltration into the retinal pigment epithelium (RPE)-choroid and suppressed the expression of inflammatory and angiogenic molecules, including vascular endothelial growth factor, monocyte chemotactic protein-1 and intercellular adhesion molecule-1. Importantly, RSV prevented the CNV-induced decrease in activated AMP-activated protein kinase and increase in activated nuclear factor-κB in the RPE-choroid complex. The regulatory effects of RSV on these molecules were confirmed in RPE, microvascular endothelial and macrophage cell lines. Inhibition of macrophage infiltration by RSV was confirmed by in vitro scratch and migration assays. RSV suppressed CNV development, reducing the levels of multiple cytokines secreted from several cell types and inhibiting macrophage migration. The direct effects of RSV on each cell type were confirmed in vitro. Although further studies are needed, RSV could potentially be applied in the clinic to prevent CNV development in AMD.

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Functional visual acuity assessment of severe atopic keratoconjunctivitis

Ibrahim Osama M A, Dogru Murat, Kaido Minako, Kojima Takashi, Fujishima Hiroshi, Tsubota Kazuo

Cornea 33 S13 - S18 2014.11

ISSN 0277-3740

PURPOSE: To evaluate changes in functional visual acuity (FVA) measurements before and after treatment with 0.1% cyclosporine ophthalmic solution for 3 months in patients with severe atopic keratoconjunctivitis (AKC).METHODS: Pairs of eyes were analyzed from 3 boys (mean age 9 ± 3.6 years) who complained of having severe AKC and were recruited for the study. Conventional Landolt visual acuity, FVA, and visual maintenance ratio (VMR) measurements were conducted before and after 3 months of 0.1% cyclosporine eye drop treatment. Tear film lipid layer interferometry, tear film break-up time, fluorescein and rose bengal vital stainings, and the Schirmer test were also performed before and after treatment.RESULTS: Eye drop treatment was associated not only with improvement of tear function and ocular surface status but also with better quality of visual function. The mean logarithm of the minimum angle of resolution FVA and Landolt visual acuity values improved from 0.45 ± 0.26 and 0.13 ± 0.17 before treatment to 0.16 ± 0.03 and -0.06 ± 0.05 after treatment, respectively. Similarly, the mean VMR values were markedly improved from 0.88 ± 0.06 to 0.95 ± 0.03 after treatment.CONCLUSIONS: FVA and VMR are promising parameters in the assessment of dynamic visual acuity changes in AKC-affected patients and evaluation of treatment outcome.

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Ocular surface reconstruction by cultivated epithelial sheet transplantation

Satake Yoshiyuki, Yamaguchi Takefumi, Hirayama Masatoshi, Higa Kazunari, Shimazaki-Den Seika, Dogru Murat, Kawakita Tetsuya, Kawashima Motoko, Shimmura Shigeto, Tsubota Kazuo, Shimazaki Jun

Cornea 33 S42 - S46 2014.11

ISSN 0277-3740

Recent advances in ocular surface reconstruction for patients with severe ocular surface diseases have significantly improved the prognosis of patients with vision-impairing corneal abnormalities. The history of cultivated epithelial sheet transplantation is short, and debate on the current approaches for these procedures is continuing. Limbal stem cell transplantation, including conjunctivolimbal autograft and keratolimbal allograft, has brought opportunities for vision improvement. In addition, the use of cultivated limbal epithelial transplantation from both allogeneic and autologous sources has provided further options for immediate postoperative epithelialization of the corneal surface. Finally, cultivated oral mucosal epithelial transplantation, which allows autologous transplantation for patients with bilateral limbal stem cell deficiency, has provided the best overall midterm and long-term results. Its biggest advantages are the absence of rejection reactions and the reduction of postoperative complications associated with steroid therapy. However, a solitary surgical approach is not sufficient for obtaining a good clinical outcome. To maximize the possibility of success using these procedures, it is important to preoperatively enhance aggressive treatment of the ocular surface, especially with factors that facilitate moisture retention. In this review article, we also discuss our clinical experience in relation to these surgical procedures.

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Clinical Backgrounds of Postoperative Keratoplasty Patients With Spontaneous Wound Dehiscence or Gaps After Suture Removal

Fujii Shota, Matsumoto Yukihiro, Fukui Masaki, Fujitake Junichi, Kawakita Tetsuya, Shimmura Shigeto, Tsubota Kazuo

Cornea 2014.10

ISSN 0277-3740

PURPOSE:: To report that spontaneous postkeratoplasty wound dehiscence or gaps caused by suture removal may occur years after penetrating and deep lamellar keratoplasty.METHODS:: We retrospectively reviewed the medical records of 667 keratoplasty patients (890 eyes). This study included 8 eyes of 8 patients (3 men and 5 women) aged 64 to 86 years who suffered from spontaneous wound dehiscence or gaps caused by suture removal. We examined their clinical backgrounds such as surgical procedure, reason for suture removal, and time interval between keratoplasty and suture removal.RESULTS:: The surgical procedures included 6 penetrating keratoplasties and 2 deep anterior lamellar keratoplasties. The most common reason for suture removal was high corneal astigmatism. Time interval between keratoplasty and suture removal was 15.9 ± 7.7 months (7–26 months). Three patients underwent suture removal from 6 months to 1 year after keratoplasty, 3 patients from 1 to 2 years, and 2 patients after more than 2 years.CONCLUSIONS:: Whenever postoperative keratoplasty suture is removed, it is important to consider that wound dehiscence or gaps may occur without incidence of trauma.

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Intraoperative and fluorescein angiographic findings of a secondary macular hole associated with age-related macular degeneration treated by pars plana vitrectomy

Okamoto Tomohiro, Shinoda Hajime, Kurihara Toshihide, Nagai Norihiro, Tsubota Kazuo, Ozawa Yoko

BMC Ophthalmology 14 ( 1 ) 2014.09

Background: Macular hole results from a tractional force at the vitreo-retinal interface which is developed by modification and subsequent degeneration of the posterior precortical vitreous and the internal limiting membrane (ILM). Meanwhile, the wet type of age-related macular degeneration (AMD) is caused by the submacular formation of choroidal neovascularization (CNV). Although exudative changes derived from CNV may cause epiretinal membrane (ERM) formation, which can also cause tractional force at the vitreo-retinal interface, there have been few reports of AMD-associated macular hole development in which the full thickness of the retinal tissue is completely torn by the tractional force. Moreover, intraoperative finding of macular hole associated with AMD with a possible involvement of subretinal lesion has not been described.Case presentation. A 78-year-old man diagnosed with wet AMD with subretinal fluid and mild cataract received 8 treatments with intravitreal pegaptanib. After AMD remission, he developed a secondary macular hole in the same eye. He underwent a pars plana vitrectomy that successfully closed the macular hole. Intraoperatively, it was found that the patient's vitreous was formed and that the ERM and ILM were adherent, suggesting the involvement of a tractional force at the vitreo-retinal interface due to an inflammatory reaction related to AMD and/or intravitreally injected chemical compounds, resulting in macular hole development. Changes in the condition of his AMD and the RPE were observed on a fluorescein angiogram (FA) and an indocyanine green angiogram (IA) that preceded macular hole development, suggesting that subretinal changes may also have been involved in the pathogenesis.Conclusion: These clinical data, including the intraoperative findings and the temporal changes in the angiograms, suggest that an inflammatory reaction at the vitreo-retinal interface and subretinal lesion related to AMD contribute to the macular hole development in AMD patients treated with intravitreal injection.

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Oral administration of royal jelly restores tear secretion capacity in rat blink-suppressed dry eye model by modulating lacrimal gland function

Imada Toshihiro, Nakamura Shigeru, Kitamura Naoki, Shibuya Izumi, Tsubota Kazuo

PLoS ONE 9 ( 9 ) 2014.09

Tears are secreted from the lacrimal gland (LG), a dysfunction in which induces dry eye, resulting in ocular discomfort and visual impairment. Honey bee products are used as a nutritional source in daily life and medicine; however, little is known about their effects on dry eye. The aim of the present study was to investigate the effects of honey bee products on tear secretion capacity in dry eye. We selected raw honey, propolis, royal jelly (RJ), pollen, or larva from commercially available honey bee products. Tear secretion capacity was evaluated following the oral administration of each honey bee product in a rat blink-suppressed dry eye model. Changes in tear secretion, LG ATP content, and LG mitochondrial levels were measured. RJ restored the tear secretion capacity and decrease in LG ATP content and mitochondrial levels to the largest extent. Royal jelly can be used as a preventative intervention for dry eye by managing tear secretion capacity in the LG. Copyright:

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First multicenter survey on infectious keratitis following excimer laser surgery in Japan

Ide Takeshi, Kurosaka Daijiro, Senoo Tadashi, Kinoshita Shigeru, Ohashi Yuichi, Inoue Yoshitsugu, Tsubota Kazuo

Taiwan Journal of Ophthalmology 4 ( 3 ) 116 - 119 2014.09

ISSN 2211-5056

Purpose: To report the first multicenter survey in Japan on infectious keratitis after excimer laser surgery. Methods: The laser in situ keratomileusis (LASIK) Safety Network (LSN) Committee sent questionnaires to 28 LSN member hospitals to assess the total number of laser corneal surgeries, the number of infection cases (including suspicious cases), and the postoperative follow-up rate during a 3-year period. Results: Responses were obtained from 27 (96.4%) of 28 institutions. One phototherapeutic keratectomy infection case was reported among 22,415 excimer laser surgery cases, which equates to an incidence rate of 0.004%. The follow-up rate was 94.14% (67.2-100%), 80.11% (41.0-96.1%), 57.95% (11.5-93.0%), and 46.64% (4.7-93.0%) at 1 month, 3 months, 6 months, and 12 months of follow-up, respectively. Conclusion: Infectious keratitis is a potentially devastating complication of excimer laser surgery. We did not see any infectious keratitis for refractive cases. This first multicenter survey in Japan on infectious keratitis provides important information on the safety of this therapy.

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Delphinidin 3,5-O-diglucoside, a constituent of the maqui berry (Aristotelia chilensis) anthocyanin, restores tear secretion in a rat dry eye model

Nakamura Shigeru, Tanaka Junji, Imada Toshihiro, Shimoda Hiroshi, Tsubota Kazuo

Journal of Functional Foods 10 346 - 354 2014.09

ISSN 1756-4646

Dry eye disease is a disorder in tear film physiology, which leads to various abnormal states of ocular surface cells. In the present study, we evaluated the potential usefulness of orally applied maqui berry (. Aristotelia chilensis) extract and the constituent anthocyanin as a preventative intervention in dry eye. Tear secretion capacity was evaluated following the oral administration of maqui berry extract to a rat blink-suppressed dry eye model. This suppressive effect of maqui berry extract and its constituent anthocyanins on reactive oxygen species formation from lacrimal gland was elevated by 2', 7'-dichlorofluorescin diacetate. Maqui berry extract and delphinidin 3,5-O-diglucoside, an anthocyanin specifically contained in the maqui berry, suppressed reactive oxygen species formation from lacrimal gland tissue and preserved tear secretion. The result of the present study demonstrates that maqui berry extract can be used as a preventative intervention for dry eye by managing tear secretion capacity in the lacrimal gland.

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Fasting mitigates immediate hypersensitivity

Nakamura Shigeru, Hisamura Ryuji, Shimoda Sachiko, Shibuya Izumi, Tsubota Kazuo

Nutrition and Metabolism 11 ( 1 ) 2014.08

Background: Fasting is a rigorous type of dietary restriction that is associate with a number of health benefits. During fasting, ketone bodies significantly increase in blood and become major body fuels, thereby sparing glucose. In the present study, we investigated effects of fasting on hypersensitivity. In addition, we also investigated the possible role of D-beta-hydroxybutyrate provoked by fasting in the attenuation of immediate hypersensitivity by fasting. Methods. Effects of fasting on systemic anaphylaxis were examined using rat model of toluene 2, 4-diisocyanate induced nasal allergy. In addition to food restriction, a ketogenic high-fat and low-carbohydrate diet that accelerates fatty acid oxidation and systemic instillation of D-beta-hydroxybutyrate were employed to elevate internal D-beta-hydroxybutyrate concentration. We assessed relationship between degranulation of rat peritoneal mast cells and internal D-beta-hydroxybutyrate concentration in each treatment. Changes in [Ca²⁺]i responses to compound 48/80 were analyzed in fura 2-loaded rat peritoneal mast cells derived from the ketogenic diet and fasting. Results: Immediate hypersensitivity reaction was significantly suppressed by fasting. A significant reduction in mast cells degranulation, induced by mast cell activator compound 48/80, was observed in rat peritoneal mast cells delivered from the 24 hours fasting treatment. In addition, mast cells delivered from a ketogenic diet and D-beta-hydroxybutyrate infusion treatment also had reduced mast cell degranulation and systemic D-beta-hydroxybutyrate concentrations were elevated to similar extent as the fasting state. The peak increase in [Ca²⁺]i was significantly lower in the ketogenic diet and fasting group than that in the control diet group. Conclusions: The results of the present study demonstrates that fasting suppress hypersensitivity reaction, and indicate that increased level of D-beta-hydroxybutyrate by fasting plays an important role, via the stabilization of mast cells, in suppression of hypersensitivity reaction.

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Rejuvenation effects of cataract surgery with ultraviolet blocking intra-ocular lens on circadian Rhythm and gait speed

Ayaki Masahiko, Negishi Kazuno, Tsubota Kazuo

Rejuvenation Research 17 ( 4 ) 359 - 365 2014.08

ISSN 1549-1684

Background: Previously, we reported improvements in sleep quality and gait speed after implantation of a yellow-colored, blue light-blocking intra-ocular lens (IOL). This study evaluated systemic health parameters for 7 months after cataract surgery with implantation of a clear, ultraviolet (UV)-blocking IOL. Methods: A total of consecutive 71 patients (average age 74.1 years) underwent cataract surgery with the implantation of a clear, UV-blocking IOL. Participants were evaluated using the Pittsburgh Sleep Quality Index (PSQI) and the National Eye Institute Visual Function Questionnaire (VFQ-25) before and at 2 and 7 months after surgery. Four-meter gait speed was also determined. The metabolic parameters of serum glycated hemoglobin (HbA1c), triglycerides (TGs), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were tested. Results: The pre-operative and post-operative (2 and 7 months after surgery) results were 66.4±16.5, 79.5±12.6, and 81.0±13.0 for VFQ-25 score, 5.7±3.5, 5.1±3.1, and 4.8±2.9 for PSQI, and 0.90±0.22, 0.91±0.22, and 0.92±0.22 meters/sec for gait speed. Significant improvements following surgery were noted in the VFQ-25 score for all cases and in the PSQI for poor sleepers (preoperative PSQI >5.5) (P<0.05, paired t-test). The gait speed and metabolic parameters showed no significant changes. Conclusions: Cataract surgery with implantation of an UV-blocking clear IOL has the potential for improving circadian rhythm and systemic health parameters. © 2014 Mary Ann Liebert, Inc.

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AMPK-NF-κB axis in the photoreceptor disorder during retinal inflammation

Kamoshita Mamoru, Ozawa Yoko, Kubota Shunsuke, Miyake Seiji, Tsuda Chiduru, Nagai Norihiro, Yuki Kenya, Shimmura Shigeto, Umezawa Kazuo, Tsubota Kazuo

PLoS ONE 9 ( 7 ) 2014.07

Recent progress in molecular analysis has revealed the possible involvement of multiple inflammatory signaling pathways in pathogenesis of retinal degeneration. However, how aberrant signaling pathways cause tissue damage and dysfunction is still being elucidated. Here, we focus on 5′-adenosine monophosphate (AMP)-activated protein kinase (AMPK), originally recognized as a key regulator of energy homeostasis. AMPK is also modulated in response to inflammatory signals, although its functions in inflamed tissue are obscure. We investigated the role of activated AMPK in the retinal neural damage and visual function impairment caused by inflammation. For this purpose, we used a mouse model of lipopolysaccharide-induced inflammation in the retina, and examined the effects of an AMPK activator, 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR). During inflammation, activated AMPK in the neural retina was decreased, but AICAR treatment prevented this change. Moreover, the electroretinogram (ERG) a-wave response, representing photoreceptor function, showed visual dysfunction in this model that was prevented by AICAR. Consistently, the model showed shortened photoreceptor outer segments (OSs) with reduced levels of rhodopsin, a visual pigment concentrated in the OSs, in a post-transcriptional manner, and these effects were also prevented by AICAR. In parallel, the level of activated NF-κB increased in the retina during inflammation, and this increase was suppressed by AICAR. Treatment with an NF-κB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ) preserved the rhodopsin level during inflammation, suppressing NF-κB. These findings indicated that AMPK activation by AICAR and subsequent NF-κB inhibition had a protective effect on visual function, and that AMPK activation played a neuroprotective role during retinal inflammation. © 2014 Kamoshita et al.

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Oxidative stress induced age dependent meibomian gland dysfunction in Cu, Zn-superoxide dismutase-1 (Sod1) knockout mice

Ibrahim Osama M A, Dogru Murat, Matsumoto Yukihiro, Igarashi Ayako, Kojima Takashi, Wakamatsu Tais Hitomi, Inaba Takaaki, Shimizu Takahiko, Shimazaki Jun, Tsubota Kazuo

PLoS ONE 9 ( 7 ) 2014.07

Purpose: The purpose of our study was to investigate alterations in the meibomian gland (MG) in Cu, Zn-Superoxide Dismutase-1 knockout (Sod1 ^-/-) mouse. Methods: Tear function tests [Break up time (BUT) and cotton thread] and ocular vital staining test were performed on Sod1 ^-/- male mice (n = 24) aged 10 and 50 weeks, and age and sex matched wild-type (+/+) mice (n = 25). Tear and serum samples were collected at sacrifice for inflammatory cytokine assays. MG specimens underwent Hematoxylin and Eosin staining, Mallory staining for fibrosis, Oil Red O lipid staining, TUNEL staining, immunohistochemistry stainings for 4HNE, 8-OHdG and CD45. Transmission electron microscopic examination (TEM) was also performed. Results: Corneal vital staining scores in the Sod1^-/- mice were significantly higher compared with the wild type mice throughout the follow-up. Tear and serum IL-6 and TNF-α levels also showed significant elevations in the 10 to 50 week Sod1^-/- mice. Oil Red O staining showed an accumulation of large lipid droplets in the Sod1^-/- mice at 50 weeks. Immunohistochemistry revealed both increased TUNEL and oxidative stress marker stainings of the MG acinar epithelium in the Sod1^-/- mice compared to the wild type mice. Immunohistochemistry staining for CD45 showed increasing inflammatory cell infiltrates from 10 to 50 weeks in the Sod1^-/- mice compared to the wild type mice. TEM revealed prominent mitochondrial changes in 50 week Sod1^-/- mice. Conclusions: Our results suggest that reactive oxygen species might play a vital role in the pathogensis of meibomian gland dysfunction. The Sod1^-/- mouse appears to be a promising model for the study of reactive oxygen species associated MG alterations. © 2014 Ibrahim et al.

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Short break-up time type dry eye has potential ocular surface abnormalities

Ichihashi Yoshiyuki, Ide Takeshi, Kaido Minako, Ishida Reiko, Hatou Shin, Tsubota Kazuo

Taiwan Journal of Ophthalmology 2014.07

ISSN 2211-5056

Purpose: To describe a case series in which corneal fluorescein staining (CFS) development occurred in short break-up time (s-BUT) dry eyes after a short period during prolonged opening of the eye. Methods: The study was designed as a clinical case series. Ocular surface evaluations were performed on 13 individuals with s-BUT dry eye. Tear function examinations included Schirmer's test and BUT evaluation. Results: In all 13 cases, the BUT was short, but the tear quantity was not so bad. In all cases, CFS developed following a single eye opening, and the staining was observed at sites that showed as dark spots. In several cases, the CFS disappeared later. Conclusions: In this study, we demonstrated that CFS could develop following a single eye opening. Based on our findings, CFS is a dynamic phenomenon rather than a stable indicator of ocular surface abnormalities. Moreover, s-BUT dry eye has the potential to show ocular surface abnormalities.

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The effects of 3% diquafosol sodium application on the tear functions and ocular surface of the Cu,Zn-superoxide dismutase-1 (Sod1)-knockout mice

Kojima Takashi, Dogru Murat, Ibrahim Osama M., Nagata Taeko, Higa Kazunari, Shimizu Takahiko, Shirasawa Takuji, Satake Yoshiyuki, Shimazaki Seika, Shimazaki Jun, Tsubota Kazuo

Molecular Vision 20 929 - 938 2014.06

To investigate the role of a water and mucin secretagogue (3% diquafosol sodium eye drops) on the tear function and conjunctival ocular surface changes in Sod1^-/- in comparison to the wild-type (WT) mice. Methods: Fourteen eyes of 7 Sod1^-/- male mice with C57BL/background and 14 eyes of 7 C57BL6 strain wild-type male mice were examined at 40 weeks in this study. All mice had application of 3% diquafosol ophthalmic solution six times a day for 2 weeks. Tear film stability and corneal epithelial damage was evaluated by fluorescein and Rose Bengal stainings. Anterior segment photography was performed before and after eye drop instillations. Aqueous tear quantity was measured with phenol red-impregnated cotton threads without anesthesia. Animals were sacrificed at 42 weeks after diquafosol treatment and the whole globe specimens were subjected to periodic acid Schiff staining. Goblet cell density was quantified by J Image software. Quantitative real-time PCR for conjunctival muc 5AC messenger RNA expression was also performed. Results: Sod1^-/- mice had significantly higher fluorescein staining scores compared to the WT mice before eye drop instillation. The mean tear film breakup time, Rose Bengal staining scores, and muc5 messenger RNA expression improved significantly with diquafosol treatment in both the WT and the knockout mice. The mean fluorescein staining score and aqueous tear quantity significantly improved in the Sod1^-/- mice with treatment. A notable and consistent increase in goblet cells and decrease in inflammatory cell infiltrates could be confirmed in all specimens after 2 weeks of diquafosol eye drop application. Conclusions: Three percent diquafosol ophthalmic solution appears to be effective in the treatment of ocular surface disease in this age-related dry eye disease mouse model. © 2014 Molecular Vision.

The use of induced pluripotent stem cells to reveal pathogenic gene mutations and explore treatments for retinitis pigmentosa

Yoshida Tetsu, Ozawa Yoko, Suzuki Keiichiro, Yuki Kenya, Ohyama Manabu, Akamatsu Wado, Matsuzaki Yumi, Shimmura Shigeto, Mitani Kohnosuke, Tsubota Kazuo, Okano Hideyuki

Molecular Brain 7 ( 1 ) 2014.06

Background: Retinitis pigmentosa (RP) is an inherited human retinal disorder that causes progressive photoreceptor cell loss, leading to severe vision impairment or blindness. However, no effective therapy has been established to date. Although genetic mutations have been identified, the available clinical data are not always sufficient to elucidate the roles of these mutations in disease pathogenesis, a situation that is partially due to differences in genetic backgrounds. Results: We generated induced pluripotent stem cells (iPSCs) from an RP patient carrying a rhodopsin mutation (E181K). Using helper-dependent adenoviral vector (HDAdV) gene transfer, the mutation was corrected in the patient's iPSCs and also introduced into control iPSCs. The cells were then subjected to retinal differentiation; the resulting rod photoreceptor cells were labeled with an Nrl promoter-driven enhanced green fluorescent protein (EGFP)-carrying adenovirus and purified using flow cytometry after 5 weeks of culture. Using this approach, we found a reduced survival rate in the photoreceptor cells with the E181K mutation, which was correlated with the increased expression of endoplasmic reticulum (ER) stress and apoptotic markers. The screening of therapeutic reagents showed that rapamycin, PP242, AICAR, NQDI-1, and salubrinal promoted the survival of the patient's iPSC-derived photoreceptor cells, with a concomitant reduction in markers of ER stress and apoptosis. Additionally, autophagy markers were found to be correlated with ER stress, suggesting that autophagy was reduced by suppressing ER stress-induced apoptotic changes. Conclusion: The use of RP patient-derived iPSCs combined with genome editing provided a versatile cellular system with which to define the roles of genetic mutations in isogenic iPSCs with or without mutation and also provided a system that can be used to explore candidate therapeutic approaches. © 2014 Yoshida et al.; licensee BioMed Central Ltd.

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Mice lacking inositol 1,4,5-trisphosphate receptors exhibit dry eye

Inaba Takaaki, Hisatsune Chihiro, Sasaki Yasumasa, Ogawa Yoko, Ebisui Etsuko, Ogawa Naoko, Matsui Minoru, Takeuchi Tsutomu, Mikoshiba Katsuhiko, Tsubota Kazuo

PLoS ONE 9 ( 6 ) 2014.06

Tear secretion is important as it supplies water to the ocular surface and keeps eyes moist. Both the parasympathetic and sympathetic pathways contribute to tear secretion. Although intracellular Ca²⁺ elevation in the acinar cells of lacrimal glands is a crucial event for tear secretion in both the pathways, the Ca²⁺ channel, which is responsible for the Ca ²⁺ elevation in the sympathetic pathway, has not been sufficiently analyzed. In this study, we examined tear secretion in mice lacking the inositol 1,4,5-trisphosphate receptor (IP₃R) types 2 and 3 (Itpr2 ^-/-;Itpr3^-/-double-knockout mice). We found that tear secretion in both the parasympathetic and sympathetic pathways was abolished in Itpr2^-/-;Itpr3^-/- mice. Intracellular Ca²⁺ elevation in lacrimal acinar cells after acetylcholine and epinephrine stimulation was abolished in Itpr2^-/-;Itpr3^-/- mice. Consequently, Itpr2^-/-;Itpr3^-/- mice exhibited keratoconjunctival alteration and corneal epithelial barrier disruption. Inflammatory cell infiltration into the lacrimal glands and elevation of serum autoantibodies, a representative marker for Sjögren's syndrome (SS) in humans, were also detected in older Itpr2^-/-;Itpr3^-/- mice. These results suggested that IP3Rs are essential for tear secretion in both parasympathetic and sympathetic pathways and that Itpr2 ^-/-;Itpr3^-/- mice could be a new dry eye mouse model with symptoms that mimic those of SS. © 2014 Inaba et al.

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Dry-eye screening by using a functional visual acuity measurement system

Kaido Minako, Uchino Miki, Yokoi Norihiko, Uchino Yuichi, Dogru Murat, Kawashima Motoko, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Kinoshita Shigeru, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 55 ( 5 ) 3275 - 3281 2014.05

ISSN 0146-0404

Purpose. We determined whether functional visual acuity (VA) parameters and a dry eyes (DEs) symptoms questionnaire could predict DEs in a population of visual terminal display (VDT) users. Methods. This prospective study included 491 VDT users from the Osaka Study. Subjects with definite DE, diagnosed with the presence of DE symptoms, tear abnormality (Schirmer test ≤ 5 mm or tear breakup time [TBUT] ≤ 5 seconds), and conjunctivocorneal epithelial damage (total staining score of ≥3 points), or probable DE, diagnosed with the presence of two of them, were assigned to a DE group, and the remainder to a non-DE group. Functional VA was assessed, and DE questionnaires were administered. We assessed whether univariate and discriminant analyses could determine to which group a subject belonged. Sensitivity and specificity were assessed. Results. Of 491 subjects, 320 and 171 were assigned to the DE and non-DE groups, respectively. No significant differences were observed between DE and non-DE groups in Schirmer test value and epithelial damage, but TBUT value (3.1 ± 1.5 vs. 5.9 ± 3.0 seconds). The sensitivity and specificity of single test using functional VA parameters were 59% and 49% in functional VA, 60% and 50% in visual maintenance ratio, and 83% and 30% in frequency of blinking, respectively. According to a discriminant analysis using a combination of functional VA parameters and a DE questionnaire, six variables were selected for the discriminant equation, of which area under the curve (AUC) was 0.735. Sensitivity and specificity of diagnoses predicted by the discriminant equation were 85.9% and 45.6%, respectively. Conclusions. The discriminant equation obtained using functional VA measurement combined with a symptoms questionnaire may suggest the possibility for the first step screening of DE with unstable tear film. Since the questionnaire has an overall poor sensitivity and specificity, further amelioration may be necessary for the actual utilization of this screening tool. © 2014 The Association for Research in Vision and Ophthalmology, Inc.

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Short-term effects of instillation of a rebamipide suspension on visual function

Kaido Minako, Ishida Reiko, Dogru Murat, Tsubota Kazuo

Journal of Ocular Pharmacology and Therapeutics 30 ( 4 ) 313 - 318 2014.05

ISSN 1080-7683

Purpose: To investigate the short-term adverse effects of using rebamipide for the treatment of dry eye by assessing visual function and optical quality. Methods: This interventional noncomparative study included 14 right eyes of 14 healthy volunteers. Serial measurements of visual acuity (VA) and higher-order aberrations were obtained prior to instillation of the rebamipide suspension (baseline) and immediately after and at 5, 10, 20, and 30 min after instillation. Functional VA measurement was performed over a 60-s period with the subject blinking naturally. Ocular aberrations were measured for 10 s while the subject was told not to blink, but no topical anesthesia was applied. Each patient also filled out a questionnaire exploring the rebamipide-Associated adverse effects. Results: There was no significant difference between functional VA measured at baseline and at each time point after the instillation of rebamipide. In contrast, the root mean square of third-order and total higher-order aberrations increased significantly immediately after drug instillation (P<0.05). The severity of higher-order aberrations at baseline was similar to that observed at 5, 10, 20, and 30 min after instillation (P>0.05). Conclusions: The transient reduction in optical quality immediately after administration of rebamipide is corrected by the patient's natural blink reflex. The adverse effects observed in this study do not outweigh the benefits of rebamipide treatment. © Copyright 2014, Mary Ann Liebert, Inc.

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Role of the Vision Van, a mobile ophthalmic outpatient clinic, in the Great East Japan Earthquake

Yuki Kenya, Nakazawa Toru, Kurosaka Daijiro, Yoshida Tsunehiko, Alfonso Eduardo C., Lee Richard K., Takano Shigeru, Tsubota Kazuo

Clinical Ophthalmology 8 691 - 696 2014.04

ISSN 1177-5467

Purpose: The Great East Japan Earthquake of March 11, 2011 triggered powerful tsunami waves off the northeastern Pacific coast of Japan that destroyed almost all of the built-up areas along the coast. The study reported here examined the role played by the Vision Van, a mobile outpatient ophthalmological clinic, in providing eye care to disaster evacuees. Methods: This was a retrospective case-series study of 2,070 victims (male: 732, female: 1,338) who visited the Vision Van. The subjects' medical records were examined retrospectively and analyzed in terms of age, sex, and date of visit to the Vision Van. Information regarding each patient's chief complaint, diagnosis, medication(s) prescribed, and eyeglasses and contact lenses provided, was also examined.Results: The Vision Van was used to conduct medical examinations on 39 days between April 23 and June 29, 2011. The average number of subjects visiting the Vision Van each day was 53±31 (range: 7-135), with examinations carried out in Miyagi Prefecture and Iwate Prefecture. The most frequent complaint was a need for eye drops (871/2,070 [42.1%]). The second and third most frequent complaints, respectively, were the need for contact lenses (294/2,070 [14.2%]) and eyeglasses (280/2,070 [13.5%]). The most frequent ocular disease diagnosis was cataract (497/2,070 [24.0%]). Eye drops were prescribed to 74.1% of the subjects.Conclusion: Mobile clinics such as the Vision Van provide valuable care, in this case, particularly to individuals who lost or left behind eyeglasses or contact lenses while escaping a natural disaster, and to subjects with chronic eye disease. © 2014 Yuki et al. This work is published by Dove Medical Press Limited.

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Phase II enzyme induction by a carotenoid, lutein, in a PC12D neuronal cell line

Miyake Seiji, Kobayashi Saori, Tsubota Kazuo, Ozawa Yoko

Biochemical and Biophysical Research Communications 446 ( 2 ) 535 - 540 2014.04

ISSN 0006-291X

The mechanism by which lutein, a carotenoid, acts as an antioxidant in retinal cells is still not fully understood. Here, lutein treatment of a neuronal cell line (PC12D) immediately resulted in reduced intracellular ROS levels, implying that it has a direct role in ROS scavenging. Significantly, lutein treatment also induced phase II antioxidative enzyme expression, probably via a nuclear factor-like 2 (Nrf2) independent pathway. This latter mechanism could explain why lutein acts diversely to protect against oxidative/cytotoxic stress, and why it is physiologically involved in the human neural tissue, such as the retina. © 2014 The Authors. Published by Elsevier Inc.

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Diclofenac protects cultured human corneal epithelial cells against hyperosmolarity and ameliorates corneal surface damage in a rat model of dry eye

Sawazaki Ryoichi, Ishihara Tomoaki, Usui Shinya, Hayashi Erika, Tahara Kayoko, Hoshino Tatsuya, Higuchi Akihiro, Nakamura Shigeru, Tsubota Kazuo, Mizushima Tohru

Investigative Ophthalmology and Visual Science 55 ( 4 ) 2547 - 2556 2014.03

ISSN 0146-0404

Purpose. Dry eye syndrome (DES) is characterized by an increase in tear osmolarity and induction of the expression and nuclear localization of an osmoprotective transcription factor (nuclear factor of activated T-cells 5 [NFAT5]) that plays an important role in providing protection against hyperosmotic tears. In this study, we screened medicines already in clinical use with a view of finding compounds that protect cultured human corneal epithelial cells against hyperosmolarity-induced cell damage. Methods. Viable cell number was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and cellular NFAT5 level was measured by immunoblotting. The rat model for DES was developed by removal of the lacrimal glands, with an assessment of corneal surface damage based on levels of fluorescein staining and epithelial apoptosis. Results. Some nonsteroidal anti-inflammatory drugs (NSAIDs), including diclofenac sodium (diclofenac), were identified during the screening procedure. These NSAIDs were able to suppress hyperosmolarity-induced apoptosis and cell growth arrest. In contrast, other NSAIDs, including bromfenac sodium (bromfenac), did not exert such a protective action. Treatment of cells with diclofenac, but not bromfenac, stimulated both the nuclear localization and expression of NFAT5 under hyperosmotic conditions. In the rat model for DES, topical administration of diclofenac (but not bromfenac) to eyes reduced corneal surface damage without affecting the volume of tear fluid. Conclusions. Diclofenac appears to protect cells against hyperosmolarity-induced cell damage and NFAT5 would play an important role in this protective action. The findings reported here may also indicate that the topical administration of diclofenac to eyes may be therapeutically beneficial for DES patients. © 2014 The Association for Research in Vision and Ophthalmology, Inc.

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Vitrectomy for myopic foveoschisis with internal limiting membrane peeling and no gas tamponade

Uchida Atsuro, Shinoda Hajime, Koto Takashi, Mochimaru Hiroshi, Nagai Norihiro, Tsubota Kazuo, Ozawa Yoko

Retina 34 ( 3 ) 455 - 460 2014.03

ISSN 0275-004X

PURPOSE:: To evaluate the outcome of vitrectomy with internal limiting membrane peeling and no gas tamponade in the treatment of eyes with myopic foveoschisis. METHODS:: Medical records of 10 eyes of 9 consecutive patients with myopic foveoschisis without macular hole treated by vitrectomy were reviewed. RESULTS:: The patientsÊ1/4 refractive error was-4.00 diopters to-34.00 diopters, and axial length was 28.38 mm to 35.90 mm. Six eyes had foveal retinal detachment with retinoschisis. All cases were treated by vitrectomy with internal limiting membrane removal without gas tamponade. The mean preoperative best-corrected visual acuity was 0.61 ± 0.42 in logarithm of the minimum angle of resolution units (Snellen equivalent of 20/82). Myopic foveoschisis was reduced in 8 eyes (80%) with a single surgery. Two eyes without improvement developed a postoperative macular hole and were treated by additional vitreoretinal surgery. All 10 eyes showed anatomical repair, and 5 eyes showed improvement in best-corrected visual acuity to 0.47 ± 0.48 (Snellen equivalent of 20/60), by 17 months after the initial surgery. CONCLUSION:: Vitrectomy with internal limiting membrane peeling and no gas tamponade can effectively treat some cases of myopic foveoschisis, suggesting that tractional forces at the vitreoretinal interface may contribute to the pathogenesis of myopic foveoschisis, thereby avoiding gas tamponade. © 2013 Ophthalmic Communications Society, Inc.

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Endoscopy-guided vitreoretinal surgery

Kawashima Shinichi, Kawashima Motoko, Tsubota Kazuo

Expert Review of Medical Devices 11 ( 2 ) 163 - 168 2014.03

ISSN 1743-4440

Vitreoretinal surgery has recently undergone remarkable developments. The introduction of micro-incision vitrectomy surgery using 25-and 23-gauge techniques has led to minimally invasive, faster, safer, and more accurate surgeries. Similarly, intraocular illumination and observation systems have advanced from light pipes to chandelier illumination, slit illumination, wide-angle viewing systems, and intraocular endoscopes. Compared to observation via the cornea and optic media, vitreous surgery using an endoscope is extremely useful, as the endoscope is inserted directly into the eye, circumventing the difficulties of corneal clouding and small pupil diameters. Furthermore, there are no blind spots after surgery. We here discuss the advantages of intraocular endoscope use, the current state of endoscopy-guided vitreoretinal surgery, and its future prospects. © 2014 Informa UK Ltd.

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Combination therapy with diquafosol tetrasodium and sodium hyaluronate in patients with dry eye after laser in situ keratomileusis

Toda Ikuko, Ide Takeshi, Fukumoto Teruki, Ichihashi Yoshiyuki, Tsubota Kazuo

American Journal of Ophthalmology 157 ( 3 ) 2014.03

ISSN 0002-9394

Purpose To evaluate the possible advantages of combination therapy with diquafosol tetrasodium and sodium hyaluronate for dry eye after laser in situ keratomileusis (LASIK). Design Prospective randomized comparative trial. Methods A total of 206 eyes of 105 patients who underwent LASIK were enrolled in this study. Patients were randomly assigned to 1 of 4 treatment groups according to the postoperative treatment: artificial tears, sodium hyaluronate, diquafosol tetrasodium, and a combination of hyaluronate and diquafosol. Questionnaire responses reflecting subjective dry eye symptoms, uncorrected and corrected visual acuity, functional visual acuity, manifest refraction, tear break-up time, fluorescein corneal staining, Schirmer test, and corneal sensitivity were examined before and 1 week and 1 month after LASIK. Results Distance uncorrected visual acuity was significantly better in the combination group than in the hyaluronate group 1 week and 1 month after LASIK. Near uncorrected visual acuity was significantly better in the combination group than in the artificial tear and diquafosol groups 1 week and 1 month after LASIK. Distance functional visual acuity improved significantly only in the combination group 1 month after LASIK. The Schirmer value in the combination group was significantly higher than that in the hyaluronate group at 1 month after LASIK. Subjective dry eye symptoms in the combination group improved significantly compared with those in the other groups 1 week after surgery. Conclusions Our results suggest that hyaluronate and diquafosol combination therapy is beneficial for early stabilization of visual performance and improvement of subjective dry eye symptoms in patients after LASIK.

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Decreased tear volume in patients with metabolic syndrome

Kawashima Motoko, Uchino Miki, Yokoi Norihiko, Dogru Murat, Uchino Yuichi, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Kinoshita Shigeru, Tsubota Kazuo

British Journal of Ophthalmology 98 ( 3 ) 418 - 420 2014.03

ISSN 0007-1161

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Association of serum lipids with macular thickness and volume in type 2 diabetes without diabetic macular edema

Sasaki Mariko, Kawashima Motoko, Kawasaki Ryo, Uchida Atsuro, Koto Takashi, Shinoda Hajime, Tsubota Kazuo, Wang Jie Jin, Ozawa Yoko

Investigative Ophthalmology and Visual Science 55 ( 3 ) 1749 - 1753 2014.02

ISSN 0146-0404

Purpose. To assess the relationship between macular thickness and volume as characterized by optical coherence tomography (OCT) and known risk factors for diabetic retinopathy (DR) or macular edema (DME) in type 2 diabetic patients with no DME. Methods. Seventy-four patients with type 2 diabetes without DME and no or only minimal DR (n = 67 with no DR; n = 7 with minimal DR; mean age, 59.5 years) were recruited at a tertiary eye hospital. Central subfield macular thickness (CSMT; circle of 500-μm radius) and central subfield macular volume (CSMV) were measured using spectral-domain OCT. Associations between OCT parameters and known risk factors for DR were examined using multiple linear regression models. Results. The mean CSMT and CSMV values were 273.7 ± 17.8 μm and 0.215 ± 0.015 mm³, respectively. After adjusting for age, sex, duration of diabetes, hemoglobin A1c, and urine protein, low-density lipoprotein (LDL) cholesterol was positively associated with CSMT and CSMV; each 1 mmol/L increase in LDL was associated with a mean increase in CSMT of 6.52 μm (95% confidence interval [CI], 1.96-11.08; P = 0.006) and a mean increase in CSMV of 0.0047 mm3 (95% CI, 0.001-0.0085; P = 0.015). Conclusions. A higher LDL cholesterol level was associated with increased CSMT and CSMV in diabetic patients without DME. Prospective longitudinal studies are warranted to assess whether having both elevated levels of LDL and higher CSMT or CSMV is a risk indicator for subsequent development of DME. © 2014 The Association for Research in Vision and Ophthalmology, Inc.

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Dry eye disease and work productivity loss in visual display users

Uchino Miki, Uchino Yuichi, Dogru Murat, Kawashima Motoko, Yokoi Norihiko, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Kinoshita Shigeru, Schaumberg Debra A., Tsubota Kazuo

American Journal of Ophthalmology 157 ( 2 ) 294 - 300 2014.02

ISSN 0002-9394

Purpose To estimate the impact of dry eye disease (DED) on work performance and productivity in office workers using visual display terminals (VDTs). Design Cross-sectional study. Methods Six hundred seventy-two Japanese young and middle-aged office workers using VDTs completed a questionnaire that was designed to measured at-work performance deficits and productivity losses using the Japanese version of the Work Limitations Questionnaire, completed by e-mail. Using the Japanese dry eye diagnostic criteria, respondents were classified into 3 groups: definite DED, probable DED, and non DED. Results Of the 672 office workers, 553 subjects (82.3%), including 366 men and 187 women, completed the questionnaire and underwent clinical evaluation. As for the total workplace productivity loss, the non DED group demonstrated a loss of 3.56%, those with probable DED demonstrated a loss of 4.06%, and those with definite DED demonstrated a loss of 4.82%, indicating significantly worse performance and productivity (P =.014, trend test). For the 4 subscales, DED was associated with significantly lower on-the-job time management (P =.009, trend test) and combined mental performance and interpersonal functioning (P =.011, trend test). After controlling for age, sex, VDT working hours, and diagnosis of DED, time management, physical demands, and mental and interpersonal functioning showed a significant relationship to DED (each P >.05). Annual DED productivity losses were estimated to be $6160 per employee when measured by total production and $1178 per employee calculated by wage. Conclusions This study indicated that there is a significant impact of DED on the total productivity of Japanese VDT users. © 2014 BY ELSEVIER INC. ALL RIGHTS RESERVED.

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Evaluation of a thermosensitive atelocollagen punctal plug treatment for dry eye disease

Kojima Takashi, Matsumoto Yukihiro, Ibrahim Osama M A, Wakamatsu Tais Hitomi, Dogru Murat, Tsubota Kazuo

American Journal of Ophthalmology 157 ( 2 ) 2014.02

ISSN 0002-9394

Purpose To evaluate the efficacy of a thermosensitive atelocollagen punctal plug in the treatment of dry eye disease. Design Prospective observational case series. Methods The thermosensitive atelocollagen punctal plug was warmed at 37 C, 39 C, 41 C, and 43 C to evaluate the appropriate temperature and time for solidification. Dry eye patients were divided into 2 groups according to the preparation method of the atelocollagen punctal plug. In the conventional implantation group, atelocollagen gel was kept at room temperature for 15 minutes before implantation (27 eyes of 14 patients). In the preheating group, atelocollagen was warmed at 41 C for 8 minutes before implantation (23 eyes of 13 dry eye patients). Strip meniscometry, vital stainings, tear film break-up time (BUT), and symptom scores were evaluated before and 1 month after plug implantation. Results In vitro experiments revealed that heating at 41 C for 8 minutes was sufficient to solidify the gel. The mean fluorescein score in the conventional implantation group significantly improved after treatment (before, 3.5 ± 2.3 points; after, 2.5 ± 0.9 points, P <.05). In the preheating group, the mean fluorescein score (before, 3.7 ± 1.7 points; after, 1.5 ± 1.2 points), strip meniscometry (before, 0.6 ± 0.7 mm; after, 1.1 ± 0.3 mm), BUT (before, 3.2 ± 0.7 seconds; after, 4.8 ± 1.0 seconds), and visual analog scale scores (before, 6.6 ± 1.5 points; after, 4.1 ± 0.9 points) significantly improved after treatment (P <.05). Conclusion The thermosensitive atelocollagen punctal plug was effective for dry eye treatment. The preheating method was found to be useful to strengthen the efficacy of the thermosensitive atelocollagen punctal plug. © 2014 BY ELSEVIER INC. ALL RIGHTS RESERVED.

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Myopic regression after phakic intraocular lens implantation and LASIK

Torii Hidemasa, Negishi Kazuno, Watanabe Kazuhiro, Arai Hiroyuki, Kato Naoko, Tsubota Kazuo

Optometry and Vision Science 91 ( 2 ) 231 - 239 2014.02

ISSN 1040-5488

PURPOSE: In myopia, biometry including the axial length is important, along with the refractive data. We compared the rates of myopic regression 3 years after phakic intraocular lens (pIOL) implantation and laser in situ keratomileusis (LASIK) after matching the preoperative axial length in highly myopic eyes of Japanese patients. METHODS: This retrospective nonrandomized study included 133 eyes of 84 patients with myopia exceeding -6.00 diopters (D) who underwent implantation of two iris-fixated pIOLs (pIOL group, 66 eyes/46 patients) or myopic LASIK (LASIK group, 67 eyes/38 patients) who were followed for more than 3 years postoperatively. The patient age, preoperative refraction, and preoperative axial length were matched between the study groups. RESULTS: There were no significant differences preoperatively between the groups in age, intraocular pressure, refraction, keratometry, or axial length. The mean regression values after 3 years compared with the 1-month postoperative refractions were -0.12 ± 0.47 (SD) D in the pIOL group and -0.82 ± 0.69 D in the LASIK group (p < 0.001). The differences in the mean regression rates between 1 and 12 months, 12 and 24 months, and 24 and 36 months of follow-up were, respectively, 0.09 ± 0.38 D, -0.11 ± 0.35 D, and -0.11 ± 0.30 D in the pIOL group and -0.57 ± 0.84 D, -0.24 ± 0.47 D, and 0.00 ± 0.53 D in the LASIK group (p < 0.001, p = 0.07, p = 0.15, respectively). CONCLUSIONS: There was a significant difference in myopic regression 3 years postoperatively between the groups matched for preoperative axial length in Japanese patients. This result has the potential to elucidate myopia in the future. Copyright © 2014 American Academy of Optometry.

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KRT12 mutations and in vivo confocal microscopy in two Japanese families with Meesmann corneal dystrophy

Ogasawara Mikihide, Matsumoto Yukihiro, Hayashi Takaaki, Ohno Kenji, Yamada Hisashi, Kawakita Tetsuya, Dogru Murat, Shimazaki Jun, Tsubota Kazuo, Tsuneoka Hiroshi

American Journal of Ophthalmology 157 ( 1 ) 2014.01

ISSN 0002-9394

Purpose To identify genetic mutations and study the corneal epithelium in Japanese patients with Meesmann corneal dystrophy. Design Laboratory investigation and prospective observational case series. Methods Slit-lamp biomicroscopy with fluorescein vital staining and in vivo confocal microscopy were performed. Mutation screening of the KRT3 and KRT12 genes was performed via polymerase chain reaction and direct sequencing for 5 patients in 2 families. Results Slit-lamp biomicroscopy revealed multiple corneal intraepithelial microcysts in all patients. A clear zone was seen in the younger generation, whereas mild subepithelial opacity was seen in the older generation. In the in vivo confocal microscopy, numerous corneal intraepithelial microcysts and hyperreflective materials, which were believed to be degenerative cells, were detected closer to the basal layer of the corneal epithelium in older patients. The superficial layer contained more enlarged microcysts, and the hyperreflective materials showed atrophic changes, as compared to the basal layer. The demarcation line between the microcysts and normal epithelial cells was clearly visualized by in vivo confocal microscopy and corresponded to the demarcation line of the clear zone observed by the slit-lamp examination. Two heterozygous mutations (Q130P, L140Q) in the KRT12 gene, one of which (L140Q) was novel, were identified only in the affected patients of the families. Conclusions We identified a novel missense mutation of the KRT12 gene in Meesmann corneal dystrophy. The in vivo confocal microscopy examinations revealed previously unreported depth-dependent ultrastructural changes in the living cornea of Meesmann corneal dystrophy patients. © 2014 by Elsevier Inc. All rights reserved.

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Clinical backgrounds of postoperative keratoplasty patients with spontaneous wound dehiscence or gaps after suture removal

Fujii Shota, Matsumoto Yukihiro, Fukui Masaki, Fujitake Junichi, Kawakita Tetsuya, Shimmura Shigeto, Tsubota Kazuo

Cornea 33 ( 12 ) 1320 - 1323 2014

ISSN 0277-3740

Purpose: To report that spontaneous postkeratoplasty wound dehiscence or gaps caused by suture removal may occur years after penetrating and deep lamellar keratoplasty. Methods: We retrospectively reviewed the medical records of 667 keratoplasty patients (890 eyes). This study included 8 eyes of 8 patients (3 men and 5 women) aged 64 to 86 years who suffered from spontaneous wound dehiscence or gaps caused by suture removal. We examined their clinical backgrounds such as surgical procedure, reason for suture removal, and time interval between keratoplasty and suture removal. Results: The surgical procedures included 6 penetrating keratoplasties and 2 deep anterior lamellar keratoplasties. The most common reason for suture removal was high corneal astigmatism. Time interval between keratoplasty and suture removal was 15.9 ± 7.7 months (7-26 months). Three patients underwent suture removal from 6 months to 1 year after keratoplasty, 3 patients from 1 to 2 years, and 2 patients after more than 2 years. Conclusions: Whenever postoperative keratoplasty suture is removed, it is important to consider that wound dehiscence or gaps may occur without incidence of trauma.

Biological effects of blocking blue and other visible light on the mouse retina

Narimatsu Toshio, Ozawa Yoko, Miyake Seiji, Kubota Shunsuke, Yuki Kenya, Nagai Norihiro, Tsubota Kazuo

Clinical and Experimental Ophthalmology 42 ( 6 ) 555 - 563 2014

ISSN 1442-6404

Background: To elucidate the biological effects of blocking fluorescent light on the retina using specific blocking materials. Methods: Seven- to 8-week-old BALB/c mice were divided into three groups and placed in one of the three boxes: one blocked ultraviolet and violet wavelengths of light (violet blockade), one blocked ultraviolet, violet, blue and some other visible wavelengths (blue-plus blockade), and one allowed most visible light to pass through (control). They were then exposed to a white fluorescent lamp for 1h at 5.65E-05mW/cm²/s. After treatment, the electroretinogram, retinal outer nuclear layer thickness and retinal outer segment length were measured. In addition, retinal apoptotic cells were quantified by TdT-mediated dUTP nick-end labelling assay and c-Fos messenger RNA, and protein levels were measured by real-time reverse-transcription polymerase chain reaction and immunoblot analyses, respectively. Results: The blue-plus blockade group retained a significantly better electroretinogram response following light exposure than the control or violet blockade groups. The blue-plus blockade group also exhibited greater outer nuclear layer thickness and greater outer-segment length, and fewer apoptotic cells after light exposure than the other groups. The c-Fos messenger RNA and protein levels were substantially reduced in the blue-plus blockade group and reduced to a lesser extent in the violet blockade group. Conclusions: The blockade of blue plus additional visible wavelengths of light was most effective in protecting the retina from light-induced damage. The blockade of violet light alone was also effective in reducing intracellular molecular responses, but these effects were not sufficient for attenuating retinal degeneration. © 2014 Royal Australian and New Zealand College of Ophthalmologists.

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Multivariable logistic regression model

Shiba Daisuke, Hatou Shin, Ono Takeshi, Hosoda Shingo, Tanabe Sachiko, Ozeki Naoki, Yuki Kenya, Shimoyama Masaru, Fukagawa Kazumi, Shimmura Shigeto, Tsubota Kazuo

PLoS ONE 9 ( 8 ) e104126 2014

PURPOSE: To design a mathematical model that can predict the relationship between the ganglion cell complex (GCC) thickness and visual field sensitivity (VFS) in glaucoma patients.DESIGN: Retrospective cross-sectional case series.METHOD: Within 3 months from VFS measurements by the Humphrey field analyzer 10-2 program, 83 eyes underwent macular GCC thickness measurements by spectral-domain optical coherence tomography (SD-OCT). Data were used to construct a multiple logistic model that depicted the relationship between the explanatory variables (GCC thickness, age, sex, and spherical equivalent of refractive errors) determined by a regression analysis and the mean VFS corresponding to the SD-OCT scanned area. Analyses were performed in half or 8 segmented local areas as well as in whole scanned areas. A simple logistic model that included GCC thickness as the single explanatory variable was also constructed. The ability of the logistic models to depict the real GCC thickness/VFS in SAP distribution was analyzed by the χ2 test of goodness-of-fit. The significance of the model effect was analyzed by analysis of variance (ANOVA).RESULTS: Scatter plots between the GCC thickness and the mean VFS showed sigmoid curves. The χ2 test of goodness-of-fit revealed that the multiple logistic models showed a good fit for the real GCC thickness/VFS distribution in all areas except the nasal-inferior-outer area. ANOVA revealed that all of the multiple logistic models significantly predicted the VFS based on the explanatory variables. Although simple logistic models also exhibited significant VFS predictability based on the GCC thickness, the model effect was less than that observed for the multiple logistic models.CONCLUSIONS: The currently proposed logistic models are useful methods for depicting relationships between the explanatory variables, including the GCC thickness, and the mean VFS in glaucoma patients.

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The association between dry eye disease and physical activity as well as sedentary behavior

Kawashima Motoko, Uchino Miki, Yokoi Norihiko, Uchino Yuichi, Dogru Murat, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Nishiwaki Yuji, Kinoshita Shigeru, Tsubota Kazuo

Journal of Ophthalmology 2014 2014

ISSN 2090-004X

Purpose. To assess the association of dry eye disease (DED) with physical activity and sedentary behavior. Methods. The cross-sectional survey conducted included Japanese office workers who use visual display terminals (n = 672). DED was assessed according to the Japanese Dry Eye Diagnostic Criteria, and participants were categorized into "definite DED," "probable DED," or "non-DED" groups based on the results of DED examinations. Physical activity and sedentary behavior of participants were assessed using the International Physical Activity Questionnaire (IPAQ), and physical activity level was calculated in metabolic equivalent units per week (MET, min/week). Participants were classified as having a high, moderate, or low level of physical activity. Results. Participants with abnormal tear break-up time (BUT) (≤5 s) were involved in sedentary behavior for significantly longer duration than those with normal BUT (P = 0. 035). Non-DED participants (14.5%) tended to have higher levels of physical activity than definite DED participants (2.5%). Participants with definite DED had significantly lower MET scores than those with non-DED (P = 0. 025). Conclusions. Our findings suggest that a lower level of physical activity and sedentary behavior are associated with DED; however, longitudinal/intervention studies with large groups of participants are needed to validate these findings.

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The effect of tinted soft contact lens wear on functional visual acuity and higher-order aberrations

Watanabe Kazuhiro, Kaido Minako, Ishida Reiko, Dogru Murat, Negishi Kazuno, Tsubota Kazuo

Contact Lens and Anterior Eye 37 ( 3 ) 203 - 208 2014

ISSN 1367-0484

Purpose: To investigate the differences of functional visual acuity (FVA) and high order aberrations (HOAs) in relation to tinted and clear hydrogel soft contact lens (SCL) wear. Methods: A prospective comparative study was performed in 16 eyes of 16 healthy volunteers. Dynamic visual acuity (using a FVA measurement system) and higher-order aberrations (using a wavefront sensor) were compared in subjects wearing two types of soft contact lenses: 1-day Acuvue^® (Vistakon, Jacksonville, FL) clear and the 1-day Acuvue^® Define^TM (Vistakon, Jacksonville, FL) tinted lens. The blink rates were recorded during FVA testing. The correlation between the difference of HOAs and differences in FVA values was analyzed. Results: The mean LogMAR FVA scores with clear and tinted SCLs were 0.07 ± 0.13 and 0.14 ± 0.17 (P< 0.05). The mean blink frequencies with clear and tinted SCL wear were 18.4 ± 8.3 and 25.3 ± 4.7. blinks/min (P< 0.05). Both 3rd-order aberrations and total HOAs showed statistically significant differences between the two types of soft contact lenses for 6. mm pupil measurements (P< 0.05). A significant positive linear correlation was observed between δHOAs and δLogMAR FVA for 6. mm pupil measurements (R= 0.53, P= 0.04). Conclusions: Tinted contact lens wear appears to induce a reduction in optical quality. Functional visual acuity measurement is a useful procedure to study the changes of visual performance and quality in tinted contact lens wear. © 2013 British Contact Lens Association.

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Aerobic exercise increases tear secretion in type 2 diabetic mice

Sano Kokoro, Kawashima Motoko, Ito Akiko, Inaba Takaaki, Morimoto Kohkichi, Watanabe Mitsuhiro, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 55 ( 7 ) 4287 - 4294 2014

ISSN 0146-0404

Purpose. To investigate the effects of exercise on tear secretion in type 2 diabetic mice, and to investigate the effect of the adenosine monophosphate-activated protein kinase (AMPK) activator 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR). Methods. Heterozygous controls (db/m; m Lepr^db) and type 2 diabetic mice (db/db; Lepr^db) either underwent forced treadmill exercise training 5 days a week or remained sedentary for 8 weeks. Tear secretion volume was measured by cotton threads for 30 seconds pre- and post intervention. The levels of oxidative stress markers (8-hydroxy-20-deoxyguanosine [8-OHdG], propanoyl lysine [PRL], and hexanoyl lysine [HEL]) in tears were measured in the postintervention period. Furthermore, C57BL/6JJc1 mice, db/db mice, and db/m mice received a single intraperitoneal injection of AICAR or PBS each day for 5 days, and tear secretion volume was measured. Results. Exercise training for 8 weeks increased tear secretion volume in db/m and db/db mice. The levels of oxidative stress markers in tears were less in the exercise group than in the control group. In C57BL/6JJc1 mice, the tear secretion volumes in both the AICAR 125 mg/kg and AICAR 250 mg/kg groups were significantly larger than in the PBS group (P < 0.01 and P < 0.01, respectively). Additionally, in db/db mice, tear secretion volume in the AICAR 125 mg/kg group was also significantly larger than in the PBS group (P < 0.05). Conclusions. Exercise training for 8 weeks and a daily injection of AICAR for 5 days increased tear secretion in mice. The results suggest that exercise may be a potential therapy to modulate tear secretion. © 2014 The Association for Research in Vision and Ophthalmology, Inc.

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Blue light injures corneal epithelial cells in the mitotic phase in vitro

Niwano Yoshimi, Kanno Taro, Iwasawa Atsuo, Ayaki Masahiko, Tsubota Kazuo

British Journal of Ophthalmology 98 ( 7 ) 990 - 992 2014

ISSN 0007-1161

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Apparent progression of presbyopia after laser in situ keratomileusis in patients with early presbyopia

Tsuneyoshi Yukari, Negishi Kazuno, Saiki Megumi, Toda Ikuko, Tsubota Kazuo

American Journal of Ophthalmology 158 ( 2 ) 286 - 292 2014

ISSN 0002-9394

Purpose To assess the effect of laser in situ keratomileusis (LASIK) on near visual function in presbyopic patients. Design Retrospective cohort study. Methods Fifty-three eyes of 40 patients 45 years of age and older who had undergone LASIK for high myopia (-6 diopters [D] and over) were included. The minimal add powers for obtaining the best-corrected near visual acuity (add powers) were measured preoperatively and 3 months postoperatively, and the correlations with the powers corrected by LASIK, corneal higher-order aberrations (HOAs), ocular HOAs, and patient ages were evaluated using univariate analysis. Factors correlated with the changes in add powers were evaluated further by multivariate analysis. Results The mean ± standard deviation patient age was 50.0 ± 4.1 years; the power corrected by LASIK was -7.56 ± 1.06 D. The mean add power was 1.80 ± 0.60 D preoperatively, which increased significantly (P <.001) to 2.18 ± 0.69 D postoperatively. Significant correlations with the increased add powers were detected with age (P =.01) and the power corrected by LASIK (P =.04) but not with corneal and ocular HOAs (P >.05). Multivariate analysis showed that only age was correlated significantly (P =.01). The percentage of eyes with increased add powers of 0.5 D or more was 60.4%, which was significantly (P =.02) higher in younger patients. Conclusions This study verified the apparent progression of presbyopia after LASIK and the importance of obtaining informed consent from patients, especially those with early presbyopia. © 2014 by Elsevier Inc. All rights reserved.

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Early signs of exudative age-related macular degeneration in Asians

Sasaki Mariko, Kawasaki Ryo, Uchida Atsuro, Koto Takashi, Shinoda Hajime, Tsubota Kazuo, Wong Tien Yin, Ozawa Yoko

Optometry and Vision Science 91 ( 8 ) 849 - 853 2014

ISSN 1040-5488

PURPOSE: To investigate the relationship between the early signs of age-related macular degeneration (AMD) and the risk of developing exudative AMD (typical AMD or polypoidal choroidal vasculopathy [PCV]) in the fellow eye of Japanese patients with unilateral exudative AMD, focusing particularly on eyes with only pigmentary abnormality. METHODS: This study is a retrospective observational consecutive case series. We retrospectively reviewed the medical charts of patients who revisited the AMD clinic from 2010 to 2011 and confirmed 129 cases with unilateral exudative AMD at their first visit (baseline). The non-affected eyes at baseline (the second eye) were categorized by the presence of early signs of AMD. The incidence of exudative AMD (typical AMD or PCV) in the fellow eye was confirmed by fluorescein and indocyanine green angiography. RESULTS: Of the 129 patients, 14 (10.9%) developed exudative AMD in the fellow eye (median follow-up, 3.2 years; n = 7 typical AMD and n = 7 PCV). Eyes with both pigmentary abnormalities and large drusen were more likely to develop typical AMD (age- and sex-adjusted odds ratio = 9.46, 95% confidence interval = 1.05 to 85.0), whereas pigmentary abnormalities without large drusen were associated with PCV (age- and sex-adjusted odds ratio = 15.9, 95% confidence interval = 1.8 to 140.5). CONCLUSIONS: There was a difference in the association between early signs of AMD and incident development of either typical AMD or PCV. Further research is warranted to determine whether pigmentary abnormalities alone may be an important risk factor for PCV in Asians. Copyright © 2014 American Academy of Optometry.

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Dry eye disease is associated with deterioration of mental health in male Japanese university staff

Tounaka Kaoru, Yuki Kenya, Kouyama Keisuke, Abe Takayuki, Tsubota Kazuo, Kawabe Hiroshi, Yokoyama Kazuhito

The Tohoku journal of experimental medicine 233 ( 3 ) 215 - 220 2014

Dry eye disease (DED) is defined as a multifactorial disease of the tears and ocular surface that results in symptoms of discomfort, visual disturbance, and tear-film instability, with potential damage to the ocular surface. It is thought to be associated with reduced quality of life (QOL). The aim of the present study was to investigate the effects of DED on health-related QOL in Japanese university sedentary office workers who are daily users of visual display terminal. In this study, 163 university staff (99 male and 64 female), aged 23-69 years, served as study subjects. Subjects were asked to answer the following three questions. (1) How often do your eyes feel dry? (2) How often do your eyes feel irritated? (3) Have you ever been diagnosed by a clinician as having dry eye syndrome? Sixty-eight subjects who answered "constantly," "often", or "sometimes" to both questions 1 and 2 were classified as the DED Group, and the remaining 95 were defined as the Non-DED Group. QOL was assessed by the SF-36 questionnaire, which consisted of 36 items to produce three summary scores, namely, mental, physical, and role/social component summary scores. For males, the DED Group had significantly lower scores than the Non-DED Group for mental component summary (MCS) (P = 0.005). In multiple regression analysis, MCS scores were adversely related to DED in males (P = 0.015). DED was associated with worsened QOL. DED should be regarded as a factor that can lead to deterioration of mental health.

[Grading for corneal endothelial damage].

Kinoshita Shigeru, Amano Shiro, Inoue Yoshitsugu, Ohashi Yuichi, Takahashi Hiroshi, Tsubota Kazuo, Nishida Kohji

Nippon Ganka Gakkai zasshi 118 ( 2 ) 81 - 83 2014

ISSN 0029-0203

While corneal endothelial damage in its most advanced stage results in bullous keratopathy where corneal transplantation is needed, there is no severity grading for corneal endothelial damage. The Japanese Corneal Society formed a working group to establish a severity grading for corneal endothelial damage. The following are the proposed severity grading criteria. Normal: Corneal endothelial cell (CEC) density is 2000 cells/mm2 or more. Grade 1: CEC density between 1000 and 2000 cells/ mm2. The corneal endothelium is below the normal physiological condition. Grade 2: CEC density between 500 and 1000 cells/ mm2. The transparency of the corneal endothelium is endangered. Even slight damage can induce bullous keratopathy. Grade 3: CEC density below 500 cells/mm2 but no corneal edema is observed. Grade 4: Bullous keratopathy. The cornea is opaque with edema.

Predictive factors for non-response to intravitreal ranibizumab treatment in age-related macular degeneration

Suzuki Misa, Nagai Norihiro, Izumi-Nagai Kanako, Shinoda Hajime, Koto Takashi, Uchida Atsuro, Mochimaru Hiroshi, Yuki Kenya, Sasaki Mariko, Tsubota Kazuo, Ozawa Yoko

British Journal of Ophthalmology 98 ( 9 ) 1186 - 1191 2014

ISSN 0007-1161

Background/aims: To study the initial characteristics and response to intravitreal ranibizumab (IVR) treatment of age-related macular degeneration (AMD). Methods: We reviewed the clinical records of 141 eyes in 141 AMD patients who received monthly IVR for 3 months and thereafter pro re nata (PRN) injections for 9 months as the first treatment for AMD. Patients whose best corrected visual acuity (BCVA) worsened at month 12, and those with increased exudative fundus findings after IVR or an increased central retinal thickness of more than 100 μm at month 12, were considered to be non-responders as judged by BCVA and fundus findings, respectively. Non-responders' initial characteristics were analysed using logistic regression models. Results: 14.9% of eyes were non-responders as judged by BCVA, and 17.0% were non-responders as judged by fundus findings. Initial fibrovascular pigment epithelial detachment (PED) (OR 22.9, 95% CI 2.61 to 201) and serous PED (OR 4.12, 95% CI 1.08 to 15.8) were associated with non-response as judged by BCVA. Initial fibrovascular PED (OR 33.5, 95% CI 2.95 to 381) and type 1 choroidal neovascularization (OR 6.46, 95% CI 1.39 to 30.0) were associated with non-response, as judged by fundus findings. Conclusions: Although most AMD responded to IVR, non-responders had initial clinical characteristics that might be informative for managing their treatment.

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Fundus autofluorescence imaging in a patient with the juvenile form of galactosialidosis

Yamazaki Risa, Tsunoda Kazushige, Fujinami Kaoru, Noda Toru, Tsubota Kazuo

Ophthalmic Surgery Lasers and Imaging Retina 45 ( 3 ) 259 - 261 2014

ISSN 2325-8160

The authors report the characteristics of fundus autofluorescence (FAF) images in a patient with galactosialidosis who presented with a macular cherry-red spot ophthalmoscopically. The cherry-red spot in the macula was hyperreflective in the FAF images. Optical coherence tomography (OCT) revealed an abnormally hyperreflective region in the retinal ganglion cell layer; however, the boundary between hyperreflective and normal regions was not clear. The findings indicate that FAF may be a more useful method to detect macular lesions than conventional funduscopic examination and OCT imaging in patients with lysosomal storage diseases presenting with a macular cherry-red spot.

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Clinical results of toric intraocular lens implantation for cataract with high astigmatism

Tominaga Takashi, Torii Hidemasa, Watanabe Kazuhiro, Shimizu Tsutomu, Saiki Megumi, Negishi Kazuno, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 68 ( 9 ) 1279 - 1283 2014

ISSN 0370-5579

Purpose: To evaluate the postoperative outcome of toric intraocular lens (TIOL) implantation for cataract patients with high corneal astigmatism. Methods: This retrospective study included 24 eyes of 22 patients with high corneal astigmatism exceeded-1.50 D who underwent cataract surgery and implantation of TIOL (SN6AT6:12 eyes, SN6AT7:8 eyes, SN6AT8:3 eyes, SN6AT9:1 eye). The mean age of all cases was 68.1 years. Visual acuity, corneal astigmatism, subjective refraction, and rotation error were recorded preoperatively and 1 month postoperatively. Results: The uncorrected and correted distance visual acuities and subjective refraction were significantly (p<0.001) improved after TIOL implantation. There was no significant difference in corneal astigmatism between preoperatively and postoperatively. The mean rotation error was 2.8°. Conclusion: TIOL was effective for cataract patients with high corneal astigmatism.

Alteration of tear mucin 5AC in office workers using visual display terminals

Uchino Yuichi, Uchino Miki, Yokoi Norihiko, Dogru Murat, Kawashima Motoko, Okada Naoko, Inaba Takaaki, Tamaki Shusaku, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Argüeso Pablo, Kinoshita Shigeru, Tsubota Kazuo

JAMA Ophthalmology 132 ( 8 ) 985 - 992 2014

ISSN 2168-6165

IMPORTANCE There are limited reports on the relationship between mucin 5AC (MUC5AC) concentrations in tears, working hours, and the frequency of ocular symptoms in visual display terminal (VDT) users. This investigation evaluated these relationships among patients with dry eye disease (DED) and individuals serving as controls. OBJECTIVE To determine the relationship between MUC5AC concentration in the tears of VDT users based on the diagnosis of DED and frequency of ocular symptoms. DESIGN, SETTING, AND PARTICIPANTS An institutional, cross-sectional studywas conducted. Participants included 96 young and middle-Aged Japanese office workers. Both eyes of 96 volunteers (60 men and 36 women) were studied. Participants working in a company that used VDTs completed questionnaires about their working hours and the frequency of ocular symptoms. Dry eye disease was diagnosed as definite or probable, or it was not present. Tear fluid was collected from the inferior fornix after instillation of 50 μL of sterilized saline. The MUC5AC concentration was normalized to tear protein content and expressed as MUC5AC (nanograms) per tear protein (milligrams). The differences in MUC5AC concentration between DED Groups, between VDT working hours (short, intermediate, and long), and between symptomatic and asymptomatic Groups were evaluated with 95%CIs based on nonparametric Hodges-Lehmann determination. MAIN OUTCOMES AND MEASURES Ocular surface evaluation, prevalence of DED, and MUC5AC concentration. RESULTS The prevalence of definite and probable DED was 9%(n = 9) and 57%(n = 55), respectively. The mean MUC5AC concentration was lower in the tears of VDT users with definite DED than in those with no DED (P = .02; Hodges-Lehmann estimator, -2.17; 95%CI, -4.67 to -0.30). The mean MUC5AC concentration in tears was lower in the Group that worked longer hours than in the Group that worked shorter hours (P = .049; estimated difference, -1.65; 95%CI, -3.12 to 0.00). Furthermore, MUC5AC concentration was lower in participants with symptomatic eye strain than in asymptomatic individuals (P = .001; estimated difference, -1.71; 95%CI, -2.86 to -0.63). CONCLUSIONS AND RELEVANCE The data obtained in the present study suggest that office workers with prolonged VDT use, as well as those with an increased frequency of eye strain, have a low MUC5AC concentration in their tears. Furthermore, MUC5AC concentration in the tears of patients with DED may be lower than that in individuals without DED. Copyright 2014 American Medical Association. All rights reserved.

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Ray tracing software for intraocular lens power calculation after corneal excimer laser surgery

Saiki Megumi, Negishi Kazuno, Kato Naoko, Torii Hidemasa, Dogru Murat, Tsubota Kazuo

Japanese Journal of Ophthalmology 58 ( 3 ) 276 - 281 2014

ISSN 0021-5155

Purpose: To evaluate the accuracy of intraocular lens (IOL) power calculations using ray tracing software in eyes after myopic laser in situ keratomileusis (LASIK). Methods: Twenty-four eyes of 17 cataract patients who underwent phacoemulsification and IOL implantation after myopic LASIK were analyzed retrospectively. The IOL power calculation was performed using OKULIX ray tracing software. The axial length was measured using the IOLMaster and keratometry data using TMS2N. The accuracy of the IOL power calculation using OKULIX was compared with those using the Camellin-Calossi, Shammas-PL, Haigis-L formulas and the double-K SRK/T formula using 43.5 diopters (D) for the Kpre. Results: The mean values of the arithmetic and absolute prediction errors were 0.63 ± 0.85 and 0.80 ± 0.68 D, respectively. The arithmetic prediction error by OKULIX was a significant hyperopic shift of the distribution of the postoperative refractive errors compared to the Camellin-Calossi, Shammas-PL and Haigis-L formulas (P < 0.05), and the absolute prediction error showed no significant difference with other formulas. The prediction errors using OKULIX were within ±0.5 D in 10 eyes (41.7 %) and within ±1.0 D in 18 eyes (75.0 %). The percentages of eyes within ±1.0 D using OKULIX were comparable to those obtained using the Camellin-Calossi, the Shammas-PL formulas and the double-K SRK/T formula using 43.5 D for the Kpre, and significantly (P < 0.05) higher than that obtained using the Haigis-L formula. Conclusions: IOL power calculations using OKULIX provided predictable outcomes in eyes that had undergone a previous myopic LASIK. © 2014 Japanese Ophthalmological Society.

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Primary and secondary surveys on epidemiology of Sjögren's syndrome in Japan

Tsuboi Hiroto, Asashima Hiromitsu, Takai Chinatsu, Hagiwara Shinya, Hagiya Chihiro, Yokosawa Masahiro, Hirota Tomoya, Umehara Hisanori, Kawakami Atsushi, Nakamura Hideki, Sano Hajime, Tsubota Kazuo, Ogawa Yoko, Takamura Etsuko, Saito Ichiro, Inoue Hiroko, Nakamura Seiji, Moriyama Masafumi, Takeuchi Tsutomu, Tanaka Yoshiya, Hirata Shintaro, Mimori Tsuneyo, Yoshifuji Hajime, Ohta Akiko, Matsumoto Isao, Sumida Takayuki

Modern Rheumatology 24 ( 3 ) 464 - 470 2014

ISSN 1439-7595

Objective. To characterize the epidemiology of Sjögren's syndrome (SS), including prevalence, disease type, extra-glandular involvement, satisfaction of diagnostic criteria sets, and treatment used in Japan. Methods. The Research Team for Autoimmune Diseases, the Research Program for Intractable Disease by the Ministry of Health, Labor and Welfare conducted primary and secondary surveys on epidemiology of SS in 2011. The primary survey covered 4,729 out of 14,095 Japan-wide Hospital Departments to investigate the prevalence of SS. The secondary survey encompassed 214 Hospital Departments that agreed to the survey, to characterize disease type, extra-glandular involvement, satisfaction of diagnostic criteria sets, and treatments. Results. The number of patients with SS in Japan estimated by the primary survey was 68,483. The secondary survey involving data collected from 2,195 SS patients from 98 Hospital Departments showed that the mean age of patients was 60.8 ± 15.2 years, male/female ratio was 1/17.4, primary/secondary SS was about 60%/40% and glandular/extra-glandular form in primary SS was about 70%/25%. The satisfaction rate was 53.8% for the 1999 revised Japanese Ministry of Health criteria for the diagnosis of SS, 47.7% for the 2002 American-European Consensus Group classification criteria for SS and 49.6% for 2012 American College of Rheumatology classification criteria for SS. Corticosteroids were used by 752 of 2,195 patients (34%), immunosuppressants by 358 patients (16%), biologics by 68 patients (3%) and secretagogues by 695 patients (32%). Conclusion. The surveys provided valuable information on the epidemiology of SS including prevalence, disease type, extra-glandular involvement, satisfaction of diagnostic criteria sets and treatments used today in Japan. © 2014 Japan College of Rheumatology.

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Possibility of measuring lutein in the retina by confocal micro-imaging system

Kudo Masahiro, Wada Satoshi, Ogawa Takayo, Namiki Sho, Ozawa Yoko, Tsubota Kazuo, Shinjo Atsushi, Suzuki Keiji

International Journal of Nanomanufacturing 10 ( 4 ) 321 - 331 2014

ISSN 1746-9392

The microfabrication and measurements of organic materials have significant potential for industrial and medical applications. In particular, in the medical field, a non-invasive method of measuring the distribution of certain substances contained in organic materials has been strongly demanded. Introducing a fluorescence analysis system to a confocal system would be useful for the characterisation of materials while maintaining space resolution. In this study, we focused on age-related macular degeneration (AMD) as an example, and investigated the possibility of the visualisation of fluorescence spectra. We measured the fluorescence spectrum in photoreceptor cells that were separated from the macular of eyes obtained from an eye-bank, using a microslicer. Mixed spectra of lutein and zeaxanthin, which are not affected by other organisations of eye have been measured. We also evaluated the densities of lutein and zeaxanthin in photoreceptor cells from the spectrum. As a result, the mixing ratio of lutein to zeaxanthin was estimated to be 3.78:6.22. The results showed good agreement with the results of high-performance liquid chromatography (HPLC). The approach presented here can serve as a useful framework for measuring the densities of lutein and zeaxanthin in photoreceptor cells.Copyright © 2014 Inderscience Enterprises Ltd.

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Association of maternal age to development and progression of retinopathy of prematurity in infants of gestational age under 33 weeks

Uchida Atsuro, Miwa Masayuki, Shinoda Hajime, Koto Takashi, Nagai Norihiro, Mochimaru Hiroshi, Tomita Yohei, Sasaki Mariko, Ikeda Kazushige, Tsubota Kazuo, Ozawa Yoko

Journal of Ophthalmology 2014 2014

ISSN 2090-004X

Aim. To find predictive and indicative markers of risk for development of retinopathy of prematurity (ROP) and its progression to the stage requiring laser treatment, in premature infants whose gestational age (GA) was under 33 weeks. Methods. We retrospectively reviewed medical records of 197 premature infants born in 2005-2010 whose G A < 33 weeks and underwent eye screening at Keio University Hospital. The association between candidate risk factors and development or progression of ROP was assessed. Results. Among the 182 eligible infants (median GA, 29.1 weeks; median birth weight (BW), 1028 g), 84 (46%) developed any stage of ROP, of which 45 (25%) required laser treatment. Multivariate analysis using a stepwise method showed that GA (P = 0.002; 95% confidence interval (CI), 0.508-0.858), BW (P < 0.001; 95% CI, 0.994-0.998), and lower maternal age (P = 0.032; 95% CI, 0.819-0.991) were the risk factors for ROP development and GA (P < 0.001; 95% CI, 0.387-0.609) and lower maternal age (P = 0.012; 95% CI, 0.795-0.973) were for laser treatment. The odds ratio of requiring laser treatment was 3.3 when the maternal age was <33 years. Conclusion. ROP was more likely to be developed and progressed in infants born from younger mother and low GA. © 2014 Atsuro Uchida et al.

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International chronic ocular graft-vs-host-disease (GVHD) consensus group

Ogawa Yoko, Kim Stella K., Dana Reza, Clayton Janine, Jain Sandeep, Rosenblatt Mark I., Perez Victor L., Shikari Hasanain, Riemens Anjo, Tsubota Kazuo

Scientific Reports 3 2013.12

The International Chronic Ocular GVHD Consensus Group held 4 working meetings to define new diagnostic metrics for chronic ocular graft-versus-host disease (GVHD). After considering the factors currently used to diagnose chronic ocular GVHD, the Consensus Group identified 4 subjective and objective variables to measure in patients following allogeneic hematopoietic stem cell transplantation (HSCT): OSDI, Schirmer's score without anesthesia, corneal staining, and conjunctival injection. Each variable was scored 0-2 or 0-3, with a maximum composite score of 11. Consideration was also given to the presence or the absence of systemic GVHD. On the basis of their composite score and the presence or absence of systemic GVHD, patients were assigned to one of three diagnostic categories: NO, PROBABLE, or DEFINITE ocular GVHD. New diagnostic criteria for chronic ocular GVHD are presented by the Consensus Group. Validation studies are needed to identify the best combination of the proposed metrics to maximize diagnostic sensitivity and specificity.

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Validation of Na,k-ATPase pump function of corneal endothelial cells for corneal regenerative medicine

Hatou Shin, Higa Kazunari, Inagaki Emi, Yoshida Satoru, Kimura Erika, Hayashi Ryuhei, Tsujikawa Motokazu, Tsubota Kazuo, Nishida Kohji, Shimmura Shigeto

Tissue Engineering - Part C: Methods 19 ( 12 ) 901 - 910 2013.12

ISSN 1937-3384

Tissue-engineering approaches to cultivate corneal endothelial cells (CECs) or induce CECs from stem cells are under investigation for the treatment of endothelial dysfunction. Before clinical application, a validation method to determine the quality of these cells is required. In this study, we quantified the endothelial pump function required for maintaining the corneal thickness using rabbit CECs (RCECs) and a human CEC line (B4G12). The potential difference of RCECs cultured on a permeable polyester membrane (Snapwell), B4G12 cells on Snapwell, or B4G12 cells on a collagen membrane (CM6) was measured by an Ussing chamber system, and the effect of different concentrations of ouabain (Na,K-ATPase specific inhibitor) was obtained. A mathematical equation derived from the concentration curve revealed that 2 mM ouabain decreases pump function of RCECs to 1.0 mV, and 0.6 mM ouabain decreases pump function of B4G12 on CM6 to 1.0 mV. Ouabain injection into the anterior chamber of rabbit eyes at a concentration of <2 mM maintained the corneal thickness, while those over 3 mM significantly increased the corneal thickness. B4G12 cell sheets transplanted into rabbit eyes treated with 0.6 mM ouabain maintained the corneal thickness, while 3.5 mM ouabain significantly increased the corneal thickness. Taken together, pump function >1.0 mV is required to maintain the corneal thickness. These results can be used for standardization of CEC pump function and validation of tissue-engineered CEC sheets for clinical use. © Copyright 2013, Mary Ann Liebert, Inc. 2013.

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Changing trends in the treatment of dry-eye disease

Dogru Murat, Nakamura Masatsugu, Shimazaki Jun, Tsubota Kazuo

Expert Opinion on Investigational Drugs 22 ( 12 ) 1581 - 1601 2013.12

ISSN 1354-3784

Introduction: Dry eye is a visually disabling disease encountered in many countries with a wide variation of treatment practices all over the world. On that front, the 2007 Report of the International Dry Eye WorkShop (DEWS) reviewed the current knowledge on all aspects of dry-eye disease (DED), in an evidence-based manner, and outlined the trends and recommendations in the treatment of DED on the basis of disease severity. Areas covered: This review mainly focuses on treatments for DED based on severity as recommended in the DEWS report, particularly artificial eye drops, hyaluronate sodium eye drops, autologous serum, anti-inflammatory eye drops including cyclosporine and steroids, and mucin secretagogues. New dry-eye treatment modalities in current trials outlined on the clinicaltrial.gov site are also outlined. Expert opinion: Further investigations into the mechanism of action of the new mucin and tear secretagogues which have been suggested to have anti-inflammatory properties will enrich our understanding in relation to relevant ocular surface responses after treatment with these new agents. © 2013 Informa UK, Ltd. © 2013 Informa UK, Ltd.

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Efficacy of a novel moist cool air device in office workers with dry eye disease

Hirayama Masatoshi, Murat Dogru, Liu Ying, Kojima Takashi, Kawakita Tetsuya, Tsubota Kazuo

Acta Ophthalmologica 91 ( 8 ) 756 - 762 2013.12

ISSN 1755-375X

Purpose: To evaluate the efficacy and safety of new moist cool air device (MCAD) for ocular symptoms, tear film stability and ocular surface status in office workers with dry eye disease (DED). Methods: In this prospective single-centre clinical trial, 40 eyes of 20 patients with DED were recruited and randomly divided into two groups (group with MCAD exposure and group without MCAD). All subjects are visual display terminals (VDTs) workers spending at least 4 h/day in front of VDTs. Patients using MCAD underwent moist air applications for 4 h/day for a total of five working weekdays during VDT works at their offices. The other group of patients performed their VDT work without moist cool air device exposure. The change in symptoms of ocular surface (OS) dryness, fatigue and discomfort was evaluated using visual analogue scale (VAS) scores. Changes in visual function, tear functions and ocular surface status were evaluated using best-corrected visual acuity (BCVA), the functional visual acuity (FVA) test, blink rate, BUT measurements, strip meniscometry (SM), tear evaporation rate, fluorescein staining and rose bengal staining scores. Tear film lipid layer interferometry was also performed to assess the status of the lipid layer over the tear film. In addition, adverse events were recorded. Results: In group with MCAD, symptoms of OS dryness during VDTs work, and FVA and BUT were significantly improved. SM and tear evaporation rate were significantly improved. There were no statistically significant differences on lipid layer stability and corneal staining scores in both groups. Blink rate was significantly increased in group without MCAD. No adverse events were reported during this trial. Conclusion: Moist cool air device use provided symptomatic relief of ocular dryness and improvement on tear stability in office workers with DED. This new device seems to be a safe and promising alternative in the treatment of DED. © 2012 The Authors. Acta Ophthalmologica © 2012 Acta Ophthalmologica Scandinavica Foundation.

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Pupillographic evaluation of relative afferent pupillary defect in glaucoma patients

Ozeki Naoki, Yuki Kenya, Shiba Daisuke, Tsubota Kazuo

British Journal of Ophthalmology 97 ( 12 ) 1538 - 1542 2013.12

ISSN 0007-1161

Aim: To evaluate the association between the magnitude of relative afferent pupillary defect (RAPD) and the laterality of visual fields in glaucoma patients using pupillography. Methods: Evaluations were made with an RAPDx (Konan Medical USA, Irvine, California, USA), which is designed to analyse pupil responses to multiple colour stimuli (white, red, green, blue, and yellow) at multiple controlled stimulus intensities. Humphrey automated perimetry (SITA standard 30-2) was performed to determine mean deviations (MDs), which were used to evaluate associations between MD differences (better eye MD - worse eye MD) and RAPDx amplitude and latency measurements in pupil response to multiple colour stimuli. Results: 58 glaucoma patients (35 men, 23 women; mean age: 62.6±12.8 years; range 28-88 years) were enrolled. The MD values were -2.00±3.1 (range -9.64 to 2.33) dB in the better eyes and -8.17±7.1 (range -26.33 to 1.07) dB in the worse eyes. The differences in MD values (better eye MD - worse eye MD) were 6.17±6.1 (range 0.05-26.41) dB. The results of linear regression analysis between the log-scaled RAPD amplitudes and differences in MD values were as follows: white: R2=0.45 (p<0.001), red: R2=0.18 (p=0.002), green: R2=0.30 (p<0.001), blue: R2=0.29 (p<0.001), and yellow: R2=0.32 (p<0.001). The log-scaled RAPD latencies and the differences in MD values were: white: R2=0.17 (p=0.001), red: R2=0.03 (p=0.22), green: R2 =0.01 (p=0.52), blue: R2=0.1 (p=0.02), and yellow: R2=0.05 (p=0.11). Conclusions: The log-scaled RAPD amplitudes correlated moderately with the differences in MD values, but the log-scaled RAPD latencies showed a weaker correlation. Stimulation with white light produced the strongest correlation with differences in MD values.

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Future direction of the anti-aging medicine

Kawashima Motoko, Tsubota Kazuo

Japanese Journal of Clinical Urology 67 ( 13 ) 1019 - 1022 2013.12

ISSN 0385-2393

A longitudinal study of Stargardt disease

Fujinami Kaoru, Lois Noemi, Mukherjee Rajarshi, McBain Vikki A., Tsunoda Kazushige, Tsubota Kazuo, Stone Edwin M., Fitzke Fred W., Bunce Catey, Moore Anthony T., Webster Andrew R., Michaelides Michel

Investigative ophthalmology & visual science 54 ( 13 ) 8181 - 8190 2013.12

We characterized subtypes of fundus autofluorescence (AF) and the progression of retinal atrophy, and correlated these findings with genotype in Stargardt disease. Full clinical examination and AF imaging was undertaken in 68 patients with Stargardt disease. The baseline data were compared to those at follow-up. Patients were classified into three AF subtypes: type 1 had a localized low signal at the fovea surrounded by a homogeneous background, type 2 had a localized low signal at the macula surrounded by a heterogeneous background with numerous foci of abnormal signal, and type 3 had multiple low signal areas at the posterior pole with a heterogeneous background. At baseline, there were 19 patients with type 1, 41 with type 2, and 8 with type 3 disease. The areas of reduced AF signal were measured and rate of atrophy enlargement (RAE) was calculated as the difference of the atrophy size over time (mm²) divided by the follow-up interval (years). Molecular screening of ABCA4 was undertaken. The mean follow-up interval was 9.1 years. A total of 42% cases with type 1 disease progressed to type 2, and 12% with type 2 progressed to type 3. The RAE (mm²/y) based upon baseline AF subtypes was significantly different; 0.06 in type 1, 0.67 in type 2, and 4.37 in type 3. ABCA4 variants were identified in 57 patients. There was a significant association between AF subtype and genotype. The AF pattern at baseline influences the enlargement of atrophy over time and has genetic correlates. These data are likely to assist in the provision of counseling on prognosis in Stargardt disease and be valuable for future clinical trials.

The association between visual field defect severity and fear of falling in primary open-angle glaucoma

Yuki Kenya, Tanabe Sachiko, Kouyama Keisuke, Fukagawa Kazumi, Uchino Miki, Shimoyama Masaru, Ozeki Naoki, Shiba Daisuke, Ozawa Yoko, Abe Takayuki, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 54 ( 12 ) 7739 - 7745 2013.11

ISSN 0146-0404

Purpose. To determine if glaucomatous visual field defect severity is associated with fear of falling. Methods. This is a cross-sectional study. A total of 387 consecutive subjects with POAG were enrolled in this study along with 293-ocular disease-free control subjects, who were screened at the same institutions. We defined mild POAG as MD of -6 dB or better, moderate POAG as MD of -6 to -12 dB, and severe POAG as MD of -12 dB or worse in the better eye. All participants were requested to answer a questionnaire on fear of falling. Associations between POAG severity and the prevalence of fear of falling were evaluated with the Cochran-Armitage trend test. Multivariable factors including age-adjusted odds ratios (ORs) for the prevalence of fear of falling and 95% confidence intervals (CIs) were evaluated with logistic regression models. Results. The p revalence of fear of falling was 35/293 (11.9%) in the control group, 38/313 (12.1%) in the mild POAG group, 12/48 (25.0%) in the moderate POAG group, and 6/26 (23.1%) in the severe POAG group, and the trend was statistically significant (P =0.028 Cochran-Armitage trend test). The adjusted ORs for prevalence in the mild, moderate, and severe POAG groups compared with that in the control group were 1.44 (95% CI: 0.83-2.51), 2.33 (95% CI: 1.00-5.44), and 4.06 (95% CI: 1.39-11.90), respectively. Conclusions. Among patients with POAG, the severity of visual field defects is associated with fear of falling. (http://www.umin.ac.jp/ctr/index.htm number, UMIN000005574.). © 2013 The Association for Research in Vision and Ophthalmology, Inc.

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The effects of 2% rebamipide ophthalmic solution on the tear functions and ocular surface of the superoxide dismutase-1 (sod1) knockout mice

Ohguchi Takeshi, Kojima Takashi, Ibrahim Osama M., Nagata Taeko, Shimizu Takahiko, Shirasawa Takuji, Kawakita Tetsuya, Satake Yoshiyuki, Tsubota Kazuo, Shimazaki Jun, Ishida Susumu

Investigative Ophthalmology and Visual Science 54 ( 12 ) 7793 - 7802 2013.11

ISSN 0146-0404

Purpose. To investigate the efficacy of 2% rebamipide ophthalmic solution on the tear functions and ocular surface status of the superoxide dismutase-1(Sod1^-/-) mice. Methods. Two percent Rebamipide ophthalmic solution was applied to 40-week-old male Sod1^-/- and wild-type (WT) mice four times a day for 2 weeks. We examined the cytokine concentrations in the tear fluid (by CytoBead assay), tear film break-up time, amount of tear production, and expressions of mucins 1, 4, and 5AC, by RT-PCR. We also performed vital staining of the ocular surface, PAS staining for muc5AC, and immunohistochemical stainings for 4-hydroxy-2-nonenal (4-HNE), 8-hydroxy-2′-deoxyguanosine (8-OHdG), in the conjunctiva to compare the Results before and after rebamipide instillations. Results. The tear functions and ocular surface epithelial damage scores were significantly worse in the Sod1^-/- than in the WT mice. Application of 2% rebamipide for 2 weeks significantly improved the tear film break-up time, the amount of tear production, and the corneal epithelial damage scores, which also significantly increased the conjunctival goblet cell density and muc5 mRNA expression, in the Sod1^-/- mice. The mean IL-6, IL-17, TNF-α, and IFN-γ levels in the tear fluid were reduced significantly along with a significant decrease in the density of cells positive for 4-HNE and 8-OHdG in the conjunctiva. Conclusions. Two percent Rebamipide ophthalmic solution significantly improved the tear stability and corneal epithelial damage, and enhanced the expression of muc5 mRNA on the ocular surface. We also observed anti-inflammatory effects in the tear film together with antioxidative effects in the conjunctiva, suggesting the efficacy of rebamipide in age-related dry eye disease attributable to SOD1 knockout. © 2013 The Association for Research in Vision and Ophthalmology, Inc.

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The clinical effect of homozygous ABCA4 ALLELES in 18 patients

Fujinami Kaoru, Sergouniotis Panagiotis I., Davidson Alice E., Mackay Donna S., Tsunoda Kazushige, Tsubota Kazuo, Robson Anthony G., Holder Graham E., Moore Anthony T., Michaelides Michel, Webster Andrew R.

Ophthalmology 120 ( 11 ) 2324 - 2331 2013.11

ISSN 0161-6420

Purpose: To describe the phenotypic presentation of a cohort of individuals with homozygous disease-associated ABCA4 variants. Design: Retrospective case series. Participants: Eighteen affected individuals from 13 families ascertained from a total cohort of 214 families with ABCA4-related retinal disease presenting to a single center. Methods: A detailed history was obtained, and color fundus photography, autofluorescence (AF) imaging, optical coherence tomography (OCT), and electrophysiologic assessment were performed. Phenotypes based on ophthalmoscopy, AF, and electrophysiology were assigned using previously reported characteristics. ABCA4 mutation detection was performed using the ABCR400 microarray (Asper Biotech, Tartu, Estonia) and high-throughput DNA sequencing, with direct sequencing used to assess segregation. Main Outcome Measures: Detailed clinical, electrophysiologic, and molecular genetic findings. Results: Eleven disease-associated homozygous ABCA4 alleles were identified, including 1 frame shift, 2 stops, 1 intronic variant causing splice-site alteration, 2 complex missense variants, and 5 missense variants: p.Glu905fsX916, p.Arg1300X, p.Gln2220X, c.4253+4 C>T, p.Leu541Pro and p.Ala1038Val (homozygosity for complex allele), p.Val931Met and p.Arg1705Gln (complex allele), p.Arg212Cys, p.Cys1488Arg, p.Arg1640Trp, p.Gly1961Glu, and p.Leu2027Phe. Eight of these 11 homozygous alleles have not been reported previously. Six of 7 patients with homozygous null alleles had early-onset (<10 years) disease, with all 7 having a severe phenotype. Two patients with homozygous missense variants (p.Leu541Pro and p.Ala1038Val [complex], and p.Arg1640Trp) presented with a severe phenotype. Three patients with homozygous p.Gly1961Glu had adult-onset disease and a mild phenotype. One patient with homozygous p.Leu2027Phe showed a spared fovea and preserved visual acuity. Conclusions: The phenotypes represented in patients identified as homozygous for presumed disease-associated ABCA4 variants gives insight into the effect of individual alleles. Null alleles have severe functional effects, and certain missense variants are similar to nulls, suggesting complete abrogation of protein function. The common alleles identified, p.Gly1961Glu and p. Leu2027Phe, both have a mild structural and functional effect on the adult retina; the latter is associated with relatively retained photoreceptor architecture and function at the fovea. Financial Disclosure(s): The author(s) have no proprietary or commercial interest in any materials discussed in this article. © 2013 by the American Academy of Ophthalmology.

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Postoperative flat anterior chamber

Ono Takeshi, Yuki Kenya, Shiba Daisuke, Abe Takayuki, Kouyama Keisuke, Tsubota Kazuo

Japanese Journal of Ophthalmology 57 ( 6 ) 520 - 528 2013.11

ISSN 0021-5155

Purpose: To investigate the long-term effects of postoperative flat anterior chamber (FAC) development on outcomes following trabeculectomy with mitomycin C. Methods: This was a retrospective cohort study. Data on 383 consecutive patients (383 eyes) who underwent trabeculectomy at our institution between 1999 and 2009 were followed up. Patients who developed FAC after trabeculectomy and patients with maintained anterior chamber were evaluated. The primary outcome variable was the success of the initial trabeculectomy, which was defined at 3 different levels by the achievement of the following intraocular pressure (IOP)-related criteria without secondary IOP-lowering surgery: (a) IOP ≤ 12 mmHg and ≤30 % reduction in IOP from the preoperative level; (b) IOP ≤ 16 mmHg and ≤20 % reduction in IOP; and (c) IOP ≤ 21 mmHg. The hazard ratios (HRs) for the failure of trabeculectomy caused by FAC within 5 years of surgery were examined in conjunction with the Cox proportional hazards regression model. Results: FAC was observed in 90 of the 383 eyes examined (23.4 %). Postoperative mild FAC was associated with the long-term success of trabeculectomy when evaluated according to our strictest success criterion, Criterion-A [HR = 0.72 (95 % CI 0.53-0.98); P = 0.04]. In contrast, severe FAC was inversely associated with the long-term success of the surgery when evaluated according to our most lenient criterion, Criterion-C [HR = 1.93 (95 % CI 1.16-3.22); P = 0.01]. Conclusion: Mild postoperative FAC after trabeculectomy is associated with a favorable long-term outcome, whereas severe postoperative FAC leads to an unfavorable prognosis. © 2013 Japanese Ophthalmological Society.

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Light-dark condition regulates sirtuin mRNA levels in the retina

Ban Norimitsu, Ozawa Yoko, Inaba Takaaki, Miyake Seiji, Watanabe Mitsuhiro, Shinmura Ken, Tsubota Kazuo

Experimental Gerontology 48 ( 11 ) 1212 - 1217 2013.11

ISSN 0531-5565

Sirtuins (Sirt1-7) are nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases/ADP-ribosyltransferases that modulate many metabolic responses affecting aging. Sirtuins expressed in tissues and organs involved in systemic metabolism have been extensively studied. However, the characteristics of sirtuins in the retina, where local energy expenditure changes dynamically in response to light stimuli, are largely unknown. Here we analyzed sirtuin mRNA levels by real-time PCR, and found that all seven sirtuins are highly expressed in the retina compared with other tissues, such as liver. We then analyzed the sirtuin mRNA profiles in the retina over time, under a 12-h light/12-h dark cycle (LD condition) and in constant darkness (DD condition). All seven sirtuins showed significant daily variation under the LD condition, with all except Sirt6 being increased in the dark phase. The expression patterns were different under the DD condition, suggesting that sirtuin mRNA levels except Sirt6 are affected by light-dark condition. These findings were not obtained in the brain and liver. In addition, the mRNA expression patterns of Nicotinamide phosphoribosyltransferase (Nampt), peroxisome proliferator-activated receptor gamma coactivator (PGC1α), and transcription factor A, mitochondrial (Tfam) in the retina, were similar to those of the sirtuins except Sirt6. Our observations provide new insights into the metabolic mechanisms of the retina and the sirtuins' regulatory systems. © 2013 The Authors.

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Keratoplasty postoperative treatment update

Shimmura-Tomita MacHiko, Shimmura Shigeto, Satake Yoshiyuki, Shimazaki-Den Seika, Omoto Masahiro, Tsubota Kazuo, Shimazaki Jun

Cornea 32 ( 11 SUPPL ) 2013.11

ISSN 0277-3740

Immunosuppressive therapy is the main postoperative treatment for keratoplasty, but there are considerable differences in protocols for the use of steroids and other immunosuppressants. Therefore, we conducted 2 prospective randomized clinical trials and 1 prospective nonrandomized clinical trial on keratoplasty postoperative treatment. One study evaluated the efficacy and safety of long-term topical corticosteroids after a penetrating keratoplasty was performed. Patients who underwent keratoplasty and maintained graft clarity for >1 year were randomly assigned to either a steroid or a no-steroid group. At the 12-month follow-up, the no-steroid group developed significantly more endothelial rejection than did the steroid group. A second study elucidated the effectiveness and safety of systemic cyclosporine in high-risk corneal transplantation. The patients were assigned to a systemic cyclosporine or control group. At a mean follow-up of 42.7 months, no difference was observed in the endothelial rejection rates and graft clarity loss between the 2 groups. A third study elucidated the effectiveness and the safety of systemic tacrolimus in high-risk corneal transplantation. Of 11 consecutive eyes decompensated despite systemic cyclosporine treatment, there was no irreversible rejection in eyes treated with tacrolimus, which was significantly better than in previous penetrating keratoplasty with systemic cyclosporine treatment. Prognosis after keratoplasty in patients with keratoconus is relatively good, but special attention is required for patients with atopic dermatitis. Postkeratoplasty atopic sclerokeratitis (PKAS) is a severe form of sclerokeratitis after keratoplasty in atopic patients. Our retrospective study showed that 35 eyes of 29 patients from a total of 247 keratoconus eyes undergoing keratoplasty were associated with atopic dermatitis, of which 6 eyes of 5 patients developed PKAS. Eyes with PKAS had a significantly higher incidence of atopic blepharitis and preoperative corneal neovascularization, and therefore, we suggest systemic corticosteroids or cyclosporine to prevent PKAS in such high-risk cases. © 2013 by Lippincott Williams & Wilkins.

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Expression and distribution of claudin subtypes in human corneal endothelium

Inagaki Emi, Hatou Shin, Yoshida Satoru, Miyashita Hideyuki, Tsubota Kazuo, Shimmura Shigeto

Investigative Ophthalmology and Visual Science 54 ( 12 ) 7258 - 7265 2013.10

ISSN 0146-0404

Purpose. To investigate the expression pattern of claudins in human corneal endothelium, and to evaluate the functional role of the claudin-10b subtype. Methods. Corneal endothelium with Descemet's membrane and the corneal epithelium were stripped from donor human corneal stroma. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the claudin subtypes expressed in corneal endothelium, stroma, and epithelium. Immunohistochemistry was performed to confirm the expression of claudin subtypes in corneal endothelium, and the expression pattern was compared to that of corneal epithelium. Finally, transendothelial resistance (TER), short-circuit current (SCC), and potential difference (PD) were measured in human corneal endothelial cell line B4G12 cells with or without claudin-10 small interfering RNA (siRNA) transfection by Ussing chamber system. Results. Transcripts for claudin-1, -2, -3, -4, -7, -10b, -11, -15, -22, -23, and -24 were identified in corneal endothelium sample by RT-PCR. Immunohistochemistry confirmed the expression of claudin-1, -2, -4, -7, -10, -11 -15, -22, and -23 in corneal endothelium. In corneal stroma, claudin-1, -2, -3, -4, -5, -6, -7, -8, -10b, -11, -12, -14, -15, -22, -23, and -24 were identified by RTPCR. In corneal epithelium, claudin-1, -3, -4, -7, -11, -14, and -23 were identified by immunohistochemistry and RT-PCR. Downregulation of claudin-10b by siRNA resulted in the decrease of SCC and PD, but not TER, in B4G12 cells. Conclusions. The expression pattern of claudin-10b⁺/claudin-14^- was specific in corneal endothelium among the three corneal layers. Claudin-10b may play an important role in the tight junction of corneal endothelium. © 2013 The Association for Research in Vision and Ophthalmology, Inc.

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Secondary implantation of piggyback intraocular lens for residual refractive error after cataract surgery

Kobayashi Fumitaka, Torii Hidemasa, Negishi Kazuno, Watanabe Kazuhiro, Saiki Megumi, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 67 ( 10 ) 1679 - 1684 2013.10

ISSN 0370-5579

Purpose: To report a case who received implantation of piggyback intraocular lens (IOL) for residual refractive error after cataract surgery. Case: A 70-year-old man presented to our clinic complaining of decreased visual acuity in his both eyes. Both eyes had received radial keratotomy 23 years before and photorefractive keratectomy 12 years before. Findings: Visual acuity was 0.5 right and 0.4 left without correction, and 0.9 right and 1.0 left with correction. Under diagnosis of age-related cataract, both eyes received phacoemulsification-aspiration with IOL implantation. Visual acuity was 0.5 in either eye without correction and 1.2 right and 1.0 left with correction one year later. The left eye had refraction of +3.00D()cyl-2.00D 65°. By request of the patient, monofocal piggyback IOL was implanted into the left eye. Six months after surgery, left visual acuity was 0.63 without and 1.0 with correction. There has been no complication throughout. Conclusion: Implantation of piggyback IOL can be a therapeutic option for residual refractive error after cataract surgery.

Calorie restriction (CR) and CR mimetics for the prevention and treatment of age-related eye disorders

Kawashima Motoko, Ozawa Yoko, Shinmura Ken, Inaba Takaaki, Nakamura Shigeru, Kawakita Tetsuya, Watanabe Mitsuhiro, Tsubota Kazuo

Experimental Gerontology 48 ( 10 ) 1096 - 1100 2013.10

ISSN 0531-5565

The morbidity of ocular diseases, including macular degeneration, diabetic retinopathy, and dry eye disease, has been gradually increasing worldwide. Because these diseases develop from age-associated ocular dysfunctions, interventions against the aging process itself may be a promising strategy for their management. Among the several approaches to interrupt aging processes, calorie restriction (CR) has been shown to recover and/or slow age-related functional declines in various organs, including the eye. Here, we review interventions against the aging process as potential therapeutic approaches to age-related ocular diseases. The effects of CR and CR mimetics in animal models of age-related eye diseases are explored. Furthermore, we discuss the possibilities of expanding this research to prospective studies to elucidate the molecular mechanisms by which CR and/or CR mimetics preserve ocular functions. © 2013 The Authors.

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Changes in higher-order aberrations after iris-fixated phakic intraocular lens implantation

Torii Hidemasa, Negishi Kazuno, Watanabe Kazuhiro, Saiki Megumi, Kato Naoko, Tsubota Kazuo

Journal of Refractive Surgery 29 ( 10 ) 693 - 700 2013.10

ISSN 1081-597X

PURPOSE: To evaluate surgically induced changes in corneal, internal, and ocular higher-order aberrations (HOAs) after phakic intraocular lens (PIOL) implantation and compare them between two PIOLs with different pupillary diameters. METHODS: Twenty-three eyes with an Artisan PIOL (Ophtec BV, Groningen, The Netherlands) (Artisan group) and 30 eyes with an Artiflex PIOL (Ophtec BV) (Artiflex group) were retrospectively evaluated. The corneal, internal, and ocular HOAs and refractive data were recorded preoperatively and 6 months postoperatively. The root mean squares (RMSs) of the total HOAs from the third- to sixth-order Zernike coefficients were calculated. The RMSs of the third- and fourth-order coefficients represented coma-like aberrations (S3) and spherical-like aberrations (S4), respectively. The main outcome measures were the differences in the parameters among the groups with different pupillary diameters. RESULTS: The mean postoperative spherical equivalents decreased from -11.84 ± 4.90 to -0.16 ± 0.40 diopters and -9.78 ± 3.20 to -0.09 ± 0.26 diopters in the Artisan and Artiflex groups, respectively. With the 4- and 5-mm pupillary diameters, the postoperative internal and ocular spherical aberrations (Z₄⁰) were significantly lower in the Artiflex group than in the Artisan group (4 mm, P = .002, .024; 5 mm, P = .004, .022, respectively). With the 6-mm pupillary diameter, there were no significant postoperative differences in any parameter measured between groups; both groups had positive spherical aberration values. CONCLUSIONS: The postoperative ocular spherical aberrations were positive after Artisan and Artiflex implantation. The postoperative ocular spherical aberration was greater in the Artisan group, possibly due to differences in the SAs of both PIOLs. Copyright © SLACK Incorporated.

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Prevalence of dry eye disease and its risk factors in visual display terminal users

Uchino Miki, Yokoi Norihiko, Uchino Yuichi, Dogru Murat, Kawashima Motoko, Komuro Aoi, Sonomura Yukiko, Kato Hiroaki, Kinoshita Shigeru, Schaumberg Debra A., Tsubota Kazuo

American Journal of Ophthalmology 156 ( 4 ) 2013.10

ISSN 0002-9394

Purpose To investigate tear function and prevalence of dry eye disease (DED) in visual display terminal (VDT) users. Design Cross-sectional study. Methods Six hundred and seventy-two young and middle-aged Japanese office workers who used VDT completed questionnaires and underwent dry eye testing. We estimated the prevalence of DED using logistic regression analysis to examine associations between DED and possible risk factors. The ocular surface feature, prevalence of DED, and risk factors were evaluated. Results Of the 672 workers, 561 (83.5%, mean age: 43.3 ± 9.1 years) completed the questionnaire. The percentage of women with a composite outcome of definite DED or probable DED was 76.5%, which was higher than that among men (60.2%; odds ratio [OR] = 2.00; 95% confidence interval [CI], 1.29-3.10, P =.002). Workers over 30 years of age had a higher risk of DED (OR = 2.22; 95% CI, 1.06-4.66), as did workers using a VDT >8 hours per day (OR = 1.94; 95% CI, 1.22-3.09). Average Schirmer value was 18.7 ± 11.7 mm and tear break-up time (TBUT) was 4.0 ± 2.5 seconds (78.6% of study participants had TBUT ≤5 seconds). Conclusions DED is prevalent among young to middle-aged Japanese VDT users. Ophthalmic findings revealed short TBUT and corneal staining accompanied by normal Schirmer test values. Increased risk for DED was noted for women aged over 30 years and prolonged VDT use. Measures to modify the adverse impact of VDT use on the ocular surface may provide a positive impact on public health and quality of life for office workers using VDTs.

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Neuroprotective role of superoxide dismutase 1 in retinal ganglioncells and inner nuclear layer cells against N-methyl-d-aspartate-induced cytotoxicity

Yuki Kenya, Yoshida Tetsu, Miyake Seiji, Tsubota Kazuo, Ozawa Yoko

Experimental Eye Research 115 230 - 238 2013.10

ISSN 0014-4835

The N-methyl- d-aspartate (NMDA) receptor-induced apoptosis is implicated in the pathological mechanisms of neural tissues, increasing the release of reactive oxygen species (ROS), resulting in a type of apoptotic cell death called excitotoxicity. Although intrinsic mechanisms to remove ROS, such as antioxidant enzymes, are provided by the tissue, the association between NMDA-induced excitotoxicity and antioxidative enzymes is not well understood. In this study, we focused on superoxide dismutase 1 (SOD1), an antioxidant enzyme, and investigated the role of SOD1 in the NMDA-induced neuronal cell death in the retina. NMDA was intravitreally injected into wild-type (WT) and SOD1 total knock-out (SOD1-deficient) mice. The number of TUNEL-positive cells in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) counted in the retinal sections and flatmount retinas were significantly higher in the SOD1-deficient mice than the WT mice after NMDA injection. Visual function assessed by dark-adapted electroretinogram (ERG) showed that the amplitudes of a-wave, b-wave, and oscillatory potential 2 were significantly reduced in the NMDA-injected SOD1-deficient mice. The level of ROS in the GCL and INL, measured using dihydroethidium, and the number of positive cells for γ-H2AX, a marker for DNA double strand breaks, and 8-OHdG, a marker for DNA oxidation, in the GCL were significantly increased in the SOD1-deficient mice after NMDA injection. We also measured mRNA and protein levels of SOD1 and SOD2 in the retina of WT mice, to find that mRNA and protein levels of SOD1, but not SOD2, were significantly reduced after NMDA injection. SOD1 deficiency exacerbated NMDA-induced damage to the inner retinal neurons, and NMDA reduced SOD1 levels in the retina of WT mice. Therefore, SOD1 protected retinal neurons against NMDA-induced retinal neurotoxicity, and NMDA-induced SOD1 reduction may be involved in neuronal vulnerability to excitotoxicity. © 2013 Elsevier Ltd.

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Long-term maintenance of limbal epithelial progenitor cells using rho kinase inhibitor and keratinocyte growth factor

Miyashita Hideyuki, Yokoo Seiichi, Yoshida Satoru, Kawakita Tetsuya, Yamagami Satoru, Tsubota Kazuo, Shimmura Shigeto

Stem Cells Translational Medicine 2 ( 10 ) 758 - 765 2013.10

ISSN 2157-6564

Corneal epithelial stem cells are located in the limbus, the junction between the cornea and the conjunctiva. A limbal epithelium model in vitro would be useful for the study of epithelial stem cells, as well as improving the quality of cultivated epithelial sheets for the treatment of limbal stem cell deficiency. In this study, we succeeded in constructing a limbal epithelium-like structure that could be maintained for at least 5 months in vitro. We modified conventional medium by replacing epidermal growth factor with keratinocyte growth factor (KGF) and adding Y-27632, a rho kinase inhibitor. Using this medium, epithelial cells freshly isolated from human limbus were cocultured with human mesenchymal stem cell-derived feeder cells. Cells formed a stratified layer without air exposure, and both basal and suprabasal layers maintained their unique morphologies for up to 5 months. Basal layers expressed the progenitor marker p63 uniformly and K15 heterogeneously. Expressions of PAX6, K3, and K12 indicated that cell sheets underwent normal differentiation in the corneal epithelium lineage. Although medium was changed daily after day 7, cell debris was observed every day, suggesting that cell sheets underwent turnover. Furthermore, secondary colonies were observed from cells dissociated from 1-month and 3-month cultured sheets. In conclusion, human limbal epithelial cell sheet cultures with KGF and Y-27632 maintained stratification, high expression of both stem/progenitor markers and differentiation markers, and colony-forming cells long-term. This protocol may be useful as an in vitro limbal epithelial model for basic studies. © AlphaMed Press 2013.

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Ophthalmic viscoelastic device injection for the treatment of flat anterior chamber after trabeculectomy

Hosoda Shingo, Yuki Kenya, Ono Takeshi, Tsubota Kazuo

Clinical Ophthalmology 7 1781 - 1785 2013.09

ISSN 1177-5467

Purpose: Flat anterior chamber (FAC) in association with overfiltration is a complication after trabeculectomy. The aim of the current study was to investigate the efficacy of injection of an ophthalmic viscoelastic device (OVD) to treat FAC after trabeculectomy in a retrospective consecutive case series study. Materials and methods: Twenty-five consecutive patients who underwent injection of OVD for the treatment of FAC after trabeculectomy were analyzed. The primary outcome variable was the qualified success of trabeculectomy, which was defined at three different levels by the achievement of one of three IOP-related criteria with or without glaucoma medication. Kaplan-Meier plots were generated to estimate survival functions for trabeculectomy with OVD injection. Results: Twenty-one patients (21/25 = 84.0%) recovered from FAC by one OVD injection. The postoperative intraocular pressure (IOP) measured at 12 months, 24 months, 36 months, 48 months, and 60 months was 14.7 ± 3.6 mmHg, 15.6 ± 7.0 mmHg, 14.2 ± 2.8 mmHg, 14.2 ± 3.8 mmHg, and 14.6 ± 2.7 mmHg, respectively. All postoperative IOP measurements were significantly lower than the preoperative IOP (P < 0.001). The probabilities of qualified success for 5 years after trabeculectomy with OVD injection were as follows: 24.4% (IOP ≦ 21 mmHg and ≧20% reduction of preoperative IOP), 16.5% (IOP ≦ 16 mmHg and ≧20% reduction) and 6.3% (IOP ≦ 12 mmHg and ≧30% reduction). One patient, who had undergone deep lamellar keratoplasty, experienced a complication associated with the injection of the viscoelastic material, Descemet's membrane detachment. Conclusion: Injection of OVD may be effective for the treatment of FAC; however, the longterm success rate was relatively low. © 2013 Hosoda et al.

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Saiki Megumi, Negishi Kazuno, Kato Naoko, Ogino Rika, Arai Hiroyuki, Toda Ikuko, Dogru Murat, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 39 ( 9 ) 1451 2013.09

ISSN 0886-3350

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Clinical and molecular analysis of stargardt disease with preserved foveal structure and function

Fujinami Kaoru, Sergouniotis Panagiotis I., Davidson Alice E., Wright Genevieve, Chana Ravinder K., Tsunoda Kazushige, Tsubota Kazuo, Egan Catherine A., Robson Anthony G., Moore Anthony T., Holder Graham E., Michaelides Michel, Webster Andrew R.

American Journal of Ophthalmology 156 ( 3 ) 2013.09

ISSN 0002-9394

Purpose: To describe a cohort of patients with Stargardt disease who show a foveal-sparing phenotype. Design: Retrospective case series. Methods: The foveal-sparing phenotype was defined as foveal preservation on autofluorescence imaging, despite a retinopathy otherwise consistent with Stargardt disease. Forty such individuals were ascertained and a full ophthalmic examination was undertaken. Following mutation screening of ABCA4, the molecular findings were compared with those of patients with Stargardt disease but no foveal sparing. Results: The median age of onset and age at examination of 40 patients with the foveal-sparing phenotype were 43.5 and 46.5 years. The median logMAR visual acuity was 0.18. Twenty-two patients (22/40, 55%) had patchy parafoveal atrophy and flecks; 8 (20%) had numerous flecks at the posterior pole without atrophy; 7 (17.5%) had mottled retinal pigment epithelial changes; 2 (5%) had multiple atrophic lesions, extending beyond the arcades; and 1 (2.5%) had a bull's-eye appearance. The median central foveal thickness assessed with spectral-domain optical coherence tomographic images was 183.0 μm (n = 33), with outer retinal tubulation observed in 15 (45%). Twenty-two of 33 subjects (67%) had electrophysiological evidence of macular dysfunction without generalized retinal dysfunction. Disease-causing variants were found in 31 patients (31/40, 78%). There was a higher prevalence of the variant p.Arg2030Gln in the cohort with foveal sparing compared to the group with foveal atrophy (6.45% vs 1.07%). Conclusions: The distinct clinical and molecular characteristics of patients with the foveal-sparing phenotype are described. The presence of 2 distinct phenotypes of Stargardt disease (foveal sparing and foveal atrophy) suggests that there may be more than 1 disease mechanism in ABCA4 retinopathy. © 2013 by Elsevier Inc. All rights reserved.

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Increased urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG)/creatinine level is associated with the progression of normal-tension glaucoma

Yuki Kenya, Tsubota Kazuo

Current Eye Research 38 ( 9 ) 983 - 988 2013.09

ISSN 0271-3683

Purpose: The objective of this study was to investigate the association between systemic oxidative stress and visual field defect progression in normal-tension glaucoma (NTG). Patients and methods: The subjects were 40 consecutive patients with NTG who were admitted to Keio University Hospital for 24-h intraocular pressure (IOP) evaluation; all subjects underwent six or more visual field tests in either eye and were followed up for >5 years. Spot samples of morning urine were collected during admission from all participants to determine the levels of urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) by ELISA kit. A linear regression line was calculated with the least squares method. Those subjects whose regression lines were negative and the p value <0.05 were classified as progressive, while all others were defined as non-progressive. Urinary 8-OHdG/creatinine level was compared between the two groups. Adjusted odds ratio and 95% confidence intervals for the progression were estimated with logistic regression models. Results: Seventeen subjects showed visual field defect progression (age: 59.9±9.5 years, untreated IOP in the right eye: 15.8±2.1mmHg), and 23 subjects showed no progression (age: 57.4±10.4 years, untreated IOP in the right eye: 16.0±2.6mmHg). Urinary 8-OHdG/creatinine level was significantly higher in the progressive group than in the non-progressive group (progressive group: 9.0±2.4ng/mg creatinine, non-progressive group: 7.3±1.8ng/mg creatinine, p=0.02). Multivariable analysis revealed that higher urinary 8-OHdG/creatinine level was a significant risk factor for the progression (odds ratio 1.54, 95% confidence interval 1.03-2.29). Conclusions: Increased urinary 8-OHdG/creatinine was associated with glaucomatous visual field progression in subjects with NTG. © Informa Healthcare USA, Inc.

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A field test of web-Based screening for dry eye disease to enhance awareness of eye problems among general internet users

Kawashima Motoko, Uchino Miki, Kawazoe Takashi, Kamiyashiki Masaaki, Sano Kokoro, Tsubota Kazuo

Journal of Medical Internet Research 15 ( 9 ) 2013.09

Background: A Web-based self-check system including a brief questionnaire would seem to be a suitable tool for rapid disease screening. Objective: The purpose of this preliminary study was to test a Web-based self-screening questionnaire for drawing attention to dry eye disease among general Internet users and identifying those with a higher risk of developing the condition. Methods: A survey website was launched and used to recruit participants from general Internet users. In the first phase, volunteers were asked to complete a Web-based self-screening questionnaire containing 12 questions on dry eye symptoms. The second phase focused on the respondents who reported five or more dry eye symptoms and expressed their intention to seek medical attention. These participants performed the Schirmer test, for evaluating tear production, and completed a paper-based lifestyle questionnaire to provide relevant background data. Results: Of the 1689 visitors to the website, 980 (58.0%) volunteers completed the Web-based self-screening questionnaire. Among these, 355 (36.2%) respondents reported five or more dry eye symptoms. Then, 99 (27.9%) of the symptomatic participants performed the Schirmer test and completed the paper-based lifestyle questionnaire. Out of these, 32 (32.2%) had abnormal tear production (≤5 mm). Conclusions: The proposed Web-based self-screening questionnaire seems to be a promising tool for raising awareness of dry eye disease among general Internet users and identifying those with a higher risk of developing the condition, although further research is needed to validate its effectiveness.

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Detection of early visual impairment in patients with epiretinal membrane

Nishi Yasuyo, Shinoda Hajime, Uchida Atsuro, Koto Takashi, Mochimaru Hiroshi, Nagai Norihiro, Tsubota Kazuo, Ozawa Yoko

Acta Ophthalmologica 91 ( 5 ) 2013.08

ISSN 1755-375X

Purpose: Patients with epiretinal membrane sometimes complain of impaired central visual function, despite good best corrected visual acuity (BCVA), as measured by visual acuity (VA) charts. Here, we evaluate early epiretinal membrane-induced changes in central VA. Methods: Subjects were 72 eyes of 36 patients with epiretinal membrane in only one eye and a BCVA in each eye better than 1.0, as measured by conventional Landolt C chart, at the Retina Division Clinic of the Department of Ophthalmology, Keio University Hospital, between December 2010 and November 2011. The conventional Landolt VA, functional VA (FVA) and contrast VA measurements were taken after a general eye examination. For the FVA, Landolt optotypes were sequentially displayed every 2 seconds, which size was changed according to the correctness of the answer. To exclude the influence of other diseases, a standard Schirmer test was performed to diagnose dry eye, and corneal and lens densities were evaluated. Results: Average BCVA measured by Landolt C chart was not changed between affected and unaffected fellow eyes. However, the affected eyes showed a poorer FVA score (0.21 ± 0.12, affected; 0.09 ± 0.12, fellow) and visual maintenance ratio (VMR) (0.90 ± 0.04, affected; 0.94 ± 0.04, fellow), measured by the FVA system, and contrast VA score (0.35 ± 0.11, affected; 0.25 ± 0.14, fellow) than fellow eyes. The FVA and contrast VA values were correlated with the presence of epiretinal membrane, but not with the presence of dry eye, cataract and corneal densities. Conclusion: FVA and contrast VA results reflected early changes in central visual function caused by epiretinal membrane, which were not detected by conventional Landolt BCVA. © 2013 The Authors. Acta Ophthalmologica © 2013 Acta Ophthalmologica Scandinavica Foundation. Published by Blackwell Publishing Ltd.

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Ocular surface complications after trabeculectomy

Ono Takeshi, Yuki Kenya, Ozeki Naoki, Shiba Daisuke, Tsubota Kazuo

Ophthalmologica 230 ( 2 ) 93 - 99 2013.08

ISSN 0030-3755

Purpose: To investigate the incidence, risk factors, time course and prognosis of ocular surface complications after trabeculectomy. Procedures: This is a retrospective, case-control study. Data on 399 consecutive patients who underwent trabeculectomy between 1999 and 2009 were evaluated. Cases of postoperative corneal epithelial defects, filamentary keratitis and dellen were evaluated. The risk factors for these ocular surface complications were analyzed with a logistic regression model. Results: Corneal epithelial defects were found in 44 of the 399 patients (11%), filamentary keratitis in 12 patients (3%) and dellen in 7 patients (2%). Dellen required surgical intervention at a statistically significantly higher rate than corneal epithelial defects or filamentary keratitis (p = 0.0001, Fisher's exact test). The significant risk factors for these ocular surface complications were found to be a history of preoperative superficial punctate keratopathy (odds ratio, OR: 2.10, 95% confidence interval, CI: 1.25-3.51, p = 0.005) and a history of diabetes mellitus (OR: 1.69, 95% CI: 1.02-2.81, p = 0.04). Conclusions: Ocular surface complications following trabeculectomy are not rare. The biggest risk factors for these complications were found to be a history of preoperative superficial punctate keratopathy and a history of diabetes mellitus. Surgery is likely to be required for the treatment of dellen. Copyright © 2013 S. Karger AG, Basel.

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Long-term results of amniotic membrane transplantation for partial limbal deficiency

Konomi Kenji, Satake Yoshiyuki, Shimmura Shigeto, Tsubota Kazuo, Shimazaki Jun

Cornea 32 ( 8 ) 1110 - 1115 2013.08

ISSN 0277-3740

PURPOSE: To evaluate the long-term efficacy of amniotic membrane transplantation as a treatment option in cases with partial limbal deficiency. METHODS: Sixteen eyes of 14 patients with partial limbal deficiency underwent amniotic membrane patch (AMP) or amniotic membrane graft with patch (AMGP) following keratectomy for conjunctivalization of the cornea. The average follow-up period was 52 months. We evaluated the mean corneal epithelialization time, visual acuity improvement, complications, and recurrence rate after the surgery. RESULTS: Eleven eyes underwent AMP, and the remaining 5 eyes received AMGP. All eyes eventually attained corneal reepithelialization. The mean time to reepithelialization was not significantly different between the AMP group (9 days) and the AMGP group (27 days). At the final follow-up visit, improved Landolt visual acuity was observed in 6 of the 11 eyes in the AMP group and in 2 of the 5 eyes in the AMGP group. No intraoperative complications were observed. However, 6 eyes in the AMP group and 3 eyes in the AMGP group had a recurrence or developed a persistent epithelial defect after mean durations of 21.2 and 17.7 months, respectively. CONCLUSION: Both AMP and AMGP appear to be effective in attaining corneal reepithelialization, but neither could provide stable, clear corneal epithelialization in the long term. Copyright © 2013 by Lippincott Williams & Wilkins.

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Saiki Megumi, Negishi Kazuno, Kato Naoko, Ogino Rika, Arai Hiroyuki, Toda Ikuko, Dogru Murat, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 39 ( 8 ) 1280 - 1281 2013.08

ISSN 0886-3350

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Molecular characteristics of four Japanese cases with KCNV2 retinopathy

Fujinami Kaoru, Tsunoda Kazushige, Nakamura Natsuko, Kato Yu, Noda Toru, Shinoda Kei, Tomita Kaoru, Hatase Tetsuhisa, Usui Tomoaki, Akahori Masakazu, Itabashi Takeshi, Iwata Takeshi, Ozawa Yoko, Tsubota Kazuo, Miyake Yozo

Molecular Vision 19 1580 - 1590 2013.07

ISSN 1090-0535

Purpose: To describe the molecular characteristics of four Japanese patients with cone dystrophy with supernormal rod responses (CDSRR). Methods: Four individuals with a clinical and electrophysiological diagnosis of CDSRR were ascertained. The pathognomonic findings of the full-field electroretinograms (ERGs) included a decrease in the rod responses, a square-shaped a-wave, an excessive increase in the b-wave in the bright flash responses, and decreased cone-derived responses. Mutational screening of the coding regions and flanking intronic sequences of the potassium channel, subfamily V, member 2 (KCNV2) gene was performed with bidirectional sequencing. The segregation of each allele was confirmed by screening other family members. Subsequent in silico analyses of the mutational consequences for protein function were performed. Results: There were two siblings from one family and one case in each of the two families. One family had a consanguineous marriage. Mutational screening revealed compound heterozygosity for the two alleles, p.C177R and p.G461R, in three patients, and homozygosity for complex alleles, p.R27H and p.R206P, in one patient from the consanguineous family. There were three putative novel variants, p.R27H, p.C177R, and p.R206P. The four variants in the families with KCNV2 were highly conserved in other species. In silico analyses predicted that all of the missense variants would alter protein function. Conclusions: Biallelic disease-causing variants were identified in four Japanese patients with CDSRR suggesting that the pathognomonic electrophysiological features are helpful in making a molecular diagnosis of KCNV2. Three novel variants were identified, and we conclude that there may be a distinct spectrum of KCNV2 alleles in the Japanese population. © 2013 Molecular Vision.

Effects of diquafosol tetrasodium administration on visual function in short break-up time dry eye

Kaido Minako, Uchino Miki, Kojima Takashi, Dogru Murat, Tsubota Kazuo

Journal of Ocular Pharmacology and Therapeutics 29 ( 6 ) 595 - 603 2013.07

ISSN 1080-7683

Purpose: To investigate the effect of diquafosol tetrasodium on tear and visual function in short break-up time (sBUT) dry eye (DE). Methods: This interventional nonrandomized comparative study involved 11 eyes in 11 patients with sBUT DE (symptom-positive sBUT group) and 13 eyes in 13 volunteers with BUT values≤5 s without DE symptoms (symptom-negative sBUT group). Tear function was assessed by measuring BUT and Schirmer values. Serial measurements were made of visual acuities (using a functional visual acuity measurement system) and higher-order aberrations (using a wavefront sensor). The parameters were compared before and 1 month after diquafosol tetrasodium instillation. Any changes in symptomatology after administration were also recorded. Results: After diquafosol tetrasodium administration, BUT values tended to increase in the symptom-positive sBUT group (P=0.07) and significantly increased in the symptom-negative sBUT group (P<0.05). LogMAR functional, minimal, and maximal visual acuities and higher-order aberrations significantly decreased after diquafosol tetrasodium administration in the symptom-positive sBUT group (P<0.05), while there were no significant changes in these values in the symptom-negative sBUT group (P>0.05). Diquafosol tetrasodium administration provided relief of DE symptoms in 75% of patients in the symptom-positive sBUT group. Conclusions: Diquafosol tetrasodium appeared to improve tear film stability and visual function in sBUT DE. © Copyright 2013, Mary Ann Liebert, Inc. 2013.

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Improvement of visual acuity after transcorneal electrical stimulation in case of Best vitelliform macular dystrophy

Ozeki Naoki, Shinoda Kei, Ohde Hisao, Ishida Susumu, Tsubota Kazuo

Graefe's Archive for Clinical and Experimental Ophthalmology 251 ( 7 ) 1867 - 1870 2013.07

ISSN 0721-832X

Purpose: To report an improvement of the visual acuity after transcorneal electrical stimulation (TES) in a case of Best vitelliform macular dystrophy (BVMD). Patient and methods: A 26-year-old woman diagnosed with BVMD presented with reduced vision. Her best corrected visual acuity (BCVA) was reduced to 20/200 in the right eye, and TES was performed once a month for two sessions. The current of the biphasic pulses (anodic first; duration, 10 msec; frequency, 20 Hz) was delivered using a DTL-electrode, and the duration of the TES was 30 min. Results: The BCVA in the right eye slowly improved after the TES, and 6 months later the BCVA was 20/25. The results of Humphrey visual field tests (VF) and multifocal ERGs (mfERGs) were only slightly changed. Two years later, the BCVA decreased, and it was improved again after another session of TES with the same parameters of the electrical pulses. Conclusion: The improvement of the visual acuity in our case of BVMD indicates that TES should be tried in other cases of retinal dystrophy. Further clinical and laboratory studies on TES are needed. © 2013 Springer-Verlag Berlin Heidelberg.

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Patient satisfaction following treatment with sodium hyaluronate or diquafosol ophthalmic solution for dry eye with short tear film break-up time

Uchino Miki, Uchino Yuichi, Fukagawa Kazumi, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 67 ( 8 ) 1403 - 1411 2013.07

ISSN 0370-5579

Purpose: To report the outcome of treatment with sodium hyaluronate or diquafosol ophthalmic solution for dry eye. Cases and Method: This study was made on 95 cases who were diagnosed with dry eye at the Ryogoku Eye Clinic, Tokyo. The series comprised 28 males and 67 females. The age ranged from 20 to 89 years. Forty-seven cases were treated with 3% diquafosol and 48 were treated with sodium hyaluronate ophthalmic solution. Patient satisfaction was evaluated after 10 weeks of treatment with 100 points as the full score. Dry eye was divided into 2 groups according to the tear break-up time (BUT). Results: Fourteen of dry eye patients with short BUT gave an average of 77.1 points after treatment with diquafosol. Eleven similar patients gave an average of 62.1 points after treatment with sodium hyaluronate. Conclusion: Higher satisfaction was obtained after treatment with diquafosol than sodium hyaluronate by dry eye patients with short BUT.

Deep stromal opacity after corneal cross-linking

Kato Naoko, Konomi Kenji, Saiki Megumi, Negishi Kazuno, Takeuchi Masaru, Shimazaki Jun, Tsubota Kazuo

Cornea 32 ( 7 ) 895 - 898 2013.07

ISSN 0277-3740

Purpose: To describe 3 cases with deep corneal stromal opacity that occurred several months after corneal cross-linking. Methods: A 36-year-old man, a 19-year-old man, and a 14-yearold girl underwent corneal cross-linking for their progressive keratoconus. Corneal cross-linking was performed according to the Dresden protocol. The corneal epithelium was ablated using an excimer laser in 2 cases and manually in 1 case. After 30 minutes of riboflavin presoaking, hypotonic riboflavin solution was applied until the corneal stroma swelled, after which the eyes were exposed to ultraviolet irradiation. Slit-lamp microscopy findings, uncorrected visual acuity, best-corrected visual acuity, manifest refraction, intraocular pressure, and corneal endothelial cell counts were evaluated, and corneal topography with Scheimpflug imaging was performed. Results: In all cases, the epithelium healed without delay. All eyes showed mild stromal infiltration a few days after the procedure; however, the inflammation was resolved within 1 week. The corneal stroma revealed no opacity up to 1 month after the procedure. A deep stromal opacity that extended to the inferior paracentral area developed after a few months and remained for 6 months to 1 year. Because the opacity was not on the visual axis, the visual acuity was not involved. Conclusions: Deep stromal opacity developed several months after uneventful corneal cross-linking. Postoperative inflammation may play a crucial role in its pathogenesis. © 2013 by Lippincott Williams & Wilkins.

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The impact of tear functions on visual outcome following keratoplasty in eyes with keratoconus

Hara Shuya, Kojima Takashi, Dogru Murat, Uchino Yuichi, Goto Eiki, Matsumoto Yukihiro, Kawakita Tetsuya, Tsubota Kazuo, Shimazaki Jun

Graefe's Archive for Clinical and Experimental Ophthalmology 251 ( 7 ) 1763 - 1770 2013.07

ISSN 0721-832X

Background: To investigate the impact of preoperative and postoperative tear functions on visual outcome and the recovery of visual function following keratoplasty in eyes with keratoconus. Methods: Twenty-five eyes of 25 consecutive patients (5 females, 20 males, mean age: 34.3 ± 15.8 years (range: 19-70 years) with keratoconus who underwent either penetrating keratoplasty (PKP) or deep anterior lamellar keratoplasty (DALK) were included in this prospective study. One patient who had a rejection episode during the follow-up period was excluded. All subjects underwent best corrected visual acuity (BCVA), corneal topography (refractive cylinder, surface regularity index: SRI and surface asymmetry index: SAI), Schirmer I test, tear film break-up time (BUT), corneal sensitivity, and fluorescein staining score measurements before as well as 1, 3, 6 and 12 months after keratoplasty. The relation between the tear functions and the duration until the achievement of maximum BCVA was also evaluated. Linear regression analysis was performed to study the statistical significance of the correlations and the time wise change of each examined parameter. Results: SRI, SAI, and BCVA significantly improved after keratoplasty. The postoperative corneal sensitivity was still significantly low even at 12 months postoperatively. Postoperative BCVA at 3, 6 months and maximum BCVA showed a significant linear negative correlation with the preoperative BUT (3 months; r = -0.461, p = 0.036, 6 months; r = -0.494, p = 0.023, maximum BCVA; r = -0.473, p = 0.030). The duration until the achievement of maximum BCVA showed a significant negative correlation with the preoperative Schirmer test (r = -0.429, p = 0.036). BCVA at 1 and 3 months postoperatively showed significant linear negative correlations with BUT value at the respective periods (1 month; r = -0.665, p = 0.0036, 3 months; r = -0.580, p = 0.0059). Conclusion: Preoperative tear functions appeared to have an influence on postoperative BCVA and the duration of visual recovery in keratoconus patients undergoing keratoplasty. Postoperative tear film stability appears to play an important role for obtaining a better BCVA in the early postoperative period after keratoplasty. © 2013 Springer-Verlag Berlin Heidelberg.

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Angiotensin II Type 1 Receptor Antagonist Attenuates Lacrimal Gland, Lung, and Liver Fibrosis in a Murine Model of Chronic Graft-Versus-Host Disease

Yaguchi Saori, Ogawa Yoko, Shimmura Shigeto, Kawakita Tetsuya, Hatou Shin, Satofuka Shingo, Nakamura Shigeru, Imada Toshihiro, Miyashita Hideyuki, Yoshida Satoru, Yaguchi Tomonori, Ozawa Yoko, Mori Takehiko, Okamoto Shinichiro, Kawakami Yutaka, Ishida Susumu, Tsubota Kazuo

PLoS ONE 8 ( 6 ) 2013.06

Chronic graft-versus-host disease (cGVHD), a serious complication following allogeneic HSCT (hematopoietic stem cell transplantation), is characterized by systemic fibrosis. The tissue renin-angiotensin system (RAS) is involved in the fibrotic pathogenesis, and an angiotensin II type 1 receptor (AT1R) antagonist can attenuate fibrosis. Tissue RAS is present in the lacrimal gland, lung, and liver, and is known to be involved in the fibrotic pathogenesis of the lung and liver. This study aimed to determine whether RAS is involved in fibrotic pathogenesis in the lacrimal gland and to assess the effect of an AT1R antagonist on preventing lacrimal gland, lung, and liver fibrosis in cGVHD model mice. We used the B10.D2→BALB/c (H-2^d) MHC-compatible, multiple minor histocompatibility antigen-mismatched model, which reflects clinical and pathological symptoms of human cGVHD. First, we examined the localization and expression of RAS components in the lacrimal glands using immunohistochemistry and quantitative real-time polymerase chain reaction (PCR). Next, we administered an AT1R antagonist (valsartan; 10 mg/kg) or angiotensin II type 2 receptor (AT2R) antagonist (PD123319; 10 mg/kg) intraperitoneally into cGVHD model mice and assessed the fibrotic change in the lacrimal gland, lung, and liver. We demonstrated that fibroblasts expressed angiotensin II, AT1R, and AT2R, and that the mRNA expression of angiotensinogen was greater in the lacrimal glands of cGVHD model mice than in controls generated by syngeneic-HSCT. The inhibition experiment revealed that fibrosis of the lacrimal gland, lung, and liver was suppressed in mice treated with the AT1R antagonist, but not the AT2R antagonist. We conclude that RAS is involved in fibrotic pathogenesis in the lacrimal gland and that AT1R antagonist has a therapeutic effect on lacrimal gland, lung, and liver fibrosis in cGVHD model mice. Our findings point to AT1R antagonist as a possible target for therapeutic intervention in cGVHD. © 2013 Yaguchi et al.

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A longitudinal study of Stargardt disease

Fujinami Kaoru, Lois Noemi, Davidson Alice E., Mackay Donna S., Hogg Chris R., Stone Edwin M., Tsunoda Kazushige, Tsubota Kazuo, Bunce Catey, Robson Anthony G., Moore Anthony T., Webster Andrew R., Holder Graham E., Michaelides Michel

American Journal of Ophthalmology 155 ( 6 ) 2013.06

ISSN 0002-9394

Purpose: To investigate the clinical and electrophysiologic natural history of Stargardt disease and correlate with the genotype. Design: Cohort study of 59 patients. Methods: Clinical history, examination, and electrophysiologic assessment were undertaken in a longitudinal survey. Patients were classified into 3 groups based on electrophysiologic findings, as previously published: Group 1 had dysfunction confined to the macula; Group 2 had macular and generalized cone system dysfunction; and Group 3 had macular and both generalized cone and rod system dysfunction. At baseline, there were 27 patients in Group 1, 17 in Group 2, and 15 in Group 3. Amplitude reduction of >50% in the relevant electroretinogram (ERG) component or a peak time shift of >3 ms for the 30 Hz flicker ERG or bright flash a-wave was considered clinically significant ERG deterioration. Molecular screening of ABCA4 was undertaken. Results: The mean age at baseline was 31.7 years, with the mean follow-up interval being 10.5 years. A total of 22% of patients from Group 1 showed ERG group transition during follow-up, with 11% progressing to Group 2 and 11% to Group 3. Forty-seven percent of patients in Group 2 progressed to Group 3. There was clinically significant ERG deterioration in 54% of all subjects: 22% of Group 1, 65% of Group 2, and 100% of Group 3. At least 1 disease-causing ABCA4 variant was identified in 47 patients. Conclusions: All patients with initial rod ERG involvement demonstrated clinically significant electrophysiologic deterioration; only 20% of patients with normal full-field ERGs at baseline showed clinically significant progression. Such data assist counseling by providing more accurate prognostic information and are also highly relevant in the design, patient selection, and monitoring of potential therapeutic interventions. © 2013 by Elsevier Inc. All Rights Reserved.

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Visante optical coherence tomography and tear function test evaluation of cholinergic treatment response in patients with Sjögren syndrome

Ibrahim Osama M A, Dogru Murat, Kawashima Shinichi, Wakamatsu Tais H., Tsubota Kazuo, Fujishima Hiroshi

Cornea 32 ( 5 ) 653 - 657 2013.05

ISSN 0277-3740

Purpose: To evaluate tear meniscus changes in patients with Sjögren syndrome (SS) receiving oral pilocarpine with Visante optical coherence tomography (OCT). Methods: Eight patients with primary SS were recruited in this prospective interventional case series study. Patients received pilocarpine tablets twice a day for 3 months. Visual analog scale assessment for dry eye and dry mouth symptoms was carried out. Patients underwent OCT and slit-lamp microscopy graticule scale tear meniscus height (TMH) measurements, strip meniscometry testing, tear film breakup time measurement, fluorescein and Rose Bengal staining, and the Schirmer 1 test. The data were analyzed 1 week, 1 month, and 3 months after treatment. Mann-Whitney test was performed. Results: Visual analog scale assessment showed a significant time-wise improvement (P < 0.05). OCT and graticule scale TMH measurements significantly improved after 1 week (P < 0.05), 1 month, and 3 months of treatment (P < 0.001). Strip meniscometry, mean tear film stability, and fluorescein and Rose Bengal scores remained improved 3 months after treatment (P < 0.001), whereas Schirmer 1 test values tended to improve without statistical significance. Conclusions: Visante OCT was effective in monitoring tear meniscus changes during the course of treatment noninvasively and quickly. Oral pilocarpine seemed to be effective in improving TMH, and the signs and symptoms of dryness in patients with SS. © 2013 Lippincott Williams & Wilkins.

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Comparison of retention rates and complications of 2 different types of silicon lacrimal punctal plugs in the treatment of dry eye disease

Kaido Minako, Ishida Reiko, Dogru Murat, Tsubota Kazuo

American Journal of Ophthalmology 155 ( 4 ) 2013.04

ISSN 0002-9394

Purpose: To investigate the retention rate and complications of 2 different types of silicon lacrimal punctal plugs with the focus on the relationship to punctum size. Design: A prospective, nonrandomized, interventional, comparative study. Method: Forty-four patients with dry eye syndrome received lacrimal punctal plug insertion with the Punctal Plug F (FCI Ophthalmics) and 30 patients received plug insertion with the Super Flex plug (Eagle Vision; Softplug-Oasis Medical Inc). In total, 110 plug insertion regions were performed using the Punctal Plug F (group A) and 110 insertion regions were carried out using Super Flex plug (group B). The gauging system of Eagle Vision was used before the lacrimal punctal plug insertion in all subjects. Plug conditions (retention rates, spontaneous plug loss, and removal) and complications related to lacrimal punctal sizes were compared between the 2 groups. Results: Retention rates were 70.4% in group A and 30.1% in group B at 6 months. Spontaneous plug loss occurrence was significant with the larger lacrimal punctum size in group A (P <.05), whereas no significant differences were observed between lacrimal punctum sizes and spontaneous loss in group B. Spontaneous plug loss occurrence was significant in elderly patients in group B (P <.05). Granuloma and injector troubles were observed in 1 and 2 cases in group A, respectively. Conclusions: The Punctal Plug F seems to be more applicable for insertion into the smaller punctal sizes. Punctum size and age seem to be important factors in determining the appropriate types of lacrimal punctal plugs. © 2013 BY ELSEVIER INC. ALL RIGHTS RESERVED.

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Modified double-K method for intraocular lens power calculation after excimer laser corneal refractive surgery

Saiki Megumi, Negishi Kazuno, Kato Naoko, Ogino Rika, Arai Hiroyuki, Toda Ikuko, Dogru Murat, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 39 ( 4 ) 556 - 562 2013.04

ISSN 0886-3350

Purpose: To propose the new anterior-posterior method (A-P method) that does not require historical data to calculate intraocular lens (IOL) power after laser in situ keratomileusis (LASIK) and to compare the accuracy of the method with other IOL formulas after LASIK. Setting: Keio University Hospital, Tokyo, Japan. Design: Case series. Methods: Eyes having phacoemulsification and IOL implantation after myopic LASIK were analyzed retrospectively. The A-P method is a modification of the double-K method using the SRK/T formula in which the estimated pre-LASIK keratometry (K) power calculated from the post-LASIK posterior sagittal power in the 6.0 mm zone is used for the preoperative K value in the double-K method and the post-LASIK anterior sagittal power is used for the postoperative K value. The accuracy of the A-P method was compared with that of other formulas that do not require preoperative data and with formulas that require preoperative data. Results: The median values of the arithmetic and absolute prediction errors using the A-P method were 0.16 diopter (D) and 0.54 D, respectively. The prediction errors using the A-P method were within ±0.50 D in 46.4% of eyes and within ±1.00 D in 75.0%. The percentage of eyes within ±1.00 D of the prediction errors with the A-P method was highest with the no-history methods. Conclusion: The A-P method may be a good option for calculating IOL power in eyes having cataract surgery after LASIK when preoperative LASIK data are unavailable. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned. © 2013 ASCRS and ESCRS.

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Glossiness and Perishable Food Quality

Murakoshi Takuma, Masuda Tomohiro, Utsumi Ken, Tsubota Kazuo, Wada Yuji

PLoS ONE 8 ( 3 ) 2013.03

Background: Previous studies have reported the effects of statistics of luminance distribution on visual freshness perception using pictures which included the degradation process of food samples. However, these studies did not examine the effect of individual differences between the same kinds of food. Here we elucidate whether luminance distribution would continue to have a significant effect on visual freshness perception even if visual stimuli included individual differences in addition to the degradation process of foods. Methodology/principal findings: We took pictures of the degradation of three fishes over 3.29 hours in a controlled environment, then cropped square patches of their eyes from the original images as visual stimuli. Eleven participants performed paired comparison tests judging the visual freshness of the fish eyes at three points of degradation. Perceived freshness scores (PFS) were calculated using the Bradley-Terry Model for each image. The ANOVA revealed that the PFS for each fish decreased as the degradation time increased; however, the differences in the PFS between individual fish was larger for the shorter degradation time, and smaller for the longer degradation time. A multiple linear regression analysis was conducted in order to determine the relative importance of the statistics of luminance distribution of the stimulus images in predicting PFS. The results show that standard deviation and skewness in luminance distribution have a significant influence on PFS. Conclusions/significance: These results show that even if foodstuffs contain individual differences, visual freshness perception and changes in luminance distribution correlate with degradation time. © 2013 Murakoshi et al.

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Functional corneal endothelium derived from corneal stroma stem cells of neural crest origin by retinoic acid and Wnt/β-catenin signaling

Hatou Shin, Yoshida Satoru, Higa Kazunari, Miyashita Hideyuki, Inagaki Emi, Okano Hideyuki, Tsubota Kazuo, Shimmura Shigeto

Stem Cells and Development 22 ( 5 ) 828 - 839 2013.03

ISSN 1547-3287

Corneal endothelial dysfunction remains a major indication for corneal transplantation. Both corneal endothelial cells and stromal cells originate from the neural crest, but have distinct phenotypes and function in the adult cornea. We previously reported that stem cells isolated from the adult corneal stroma [cornea-derived precursors (COPs)] show characteristics of multipotent neural crest-derived stem cells. In this study, we report the induction of functional tissue-engineered corneal endothelium (TECE) from mouse and human COPs. TECE was engineered from Wnt1-Cre/Floxed EGFP mouse COPs in a medium containing retinoic acid and glycogen synthase kinase (GSK) 3β inhibitor (activator of Wnt/β-catenin signaling). The expression levels of major markers characterizing corneal endothelial function (Atp1a1, Slc4a4, Car2, Col4a2, Col8a2, and Cdh2) were significantly upregulated. Both retinoic acid and GSK 3β inhibitor upregulated the expression of Pitx2, a homeobox gene involved in the development of the anterior segment of the eye. GSK 3β inhibitor increased Atp1a1 expression and Na,K-ATPase pump activity of TECE, which was significantly higher than COPs or control 3T3 cells, and 2.6-fold higher than cultured mouse corneal endothelial cells. Mouse TECE transplanted into rabbit corneas maintained transparency and corneal thickness, whereas control corneas without TECE showed marked edema and increased corneal thickness. Furthermore, we successfully induced TECE from human COPs, and human TECE transplanted into rabbit corneas also maintained corneal transparency and thickness. This protocol enables efficient production of corneal endothelium from corneal stromal stem cells by direct induction, which may lead to a novel stem cell therapy for corneal endothelial dysfunction. © 2013, Mary Ann Liebert, Inc.

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A new central - Peripheral corneal curvature method for intraocular lens power calculation after excimer laser refractive surgery

Saiki Megumi, Negishi Kazuno, Kato Naoko, Arai Hiroyuki, Toda Ikuko, Torii Hidemasa, Dogru Murat, Tsubota Kazuo

Acta Ophthalmologica 91 ( 2 ) 2013.03

ISSN 1755-375X

Purpose: To propose the central-peripheral (C-P) method, which requires no data history to calculate intraocular lens (IOL) powers for eyes that underwent laser in situ keratomileusis (LASIK), and compare the accuracy of the C-P method with other IOL formulas for eyes after LASIK. Methods: Sixteen patients with cataract (25 eyes) who underwent myopic LASIK were analysed retrospectively. The C-P method is a modified double-K method using the SRK/T formula, in which the estimated pre-LASIK keratometric power calculated from the post-LASIK peripheral anterior sagittal power (also called the axial power) is used for the Kpre in the double-K method using the SRK/T formula, and the post-LASIK anterior sagittal power is used for the Kpost. We compared the accuracy of the C-P method with other popular IOL calculation formulas for use in eyes after LASIK. Results: The median values of the arithmetic and absolute prediction errors with the C-P method were 0.11 diopter (D) (range, -1.67 to 1.97 D) and 0.55 D (range, 0.02-1.97 D), respectively. The prediction error using the C-P method was within ±0.5 D in 48% of eyes, within -1.0 to +0.5 D in 60% of eyes, and within ±1.0 D in 68% of eyes. The C-P method resulted in a significantly higher percentage of eyes within ±0.5 D than the BESSt formula, Shammas-PL formula, true net power method, double-K method using 43.5 D for Kpre, and Feiz-Mannis method. Conclusion: The C-P method may be a good option for calculating IOL powers in eyes undergoing cataract surgery after LASIK when the preoperative LASIK data are unavailable. © 2013 The Authors Acta Ophthalmologica © 2013 Acta Ophthalmologica Scandinavica Foundation.

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Serum vascular adhesion protein-1 correlates with vascular endothelial growth factor in patients with type II diabetes

Yoshikawa Nami, Noda Kousuke, Shinoda Hajime, Uchida Atsuro, Ozawa Yoko, Tsubota Kazuo, Mashima Yukihiko, Ishida Susumu

Journal of Diabetes and its Complications 27 ( 2 ) 162 - 166 2013.03

ISSN 1056-8727

Aims: To study serum levels of soluble vascular adhesion protein (sVAP)-1 in type II diabetic patients with retinopathy. Methods: Serum samples were obtained from 53 consecutive patients, including 14 cases with non-angiogenic ocular diseases, i.e., epiretinal membrane (ERM) and idiopathic macular hole (MH), 19 cases with age-related macular degeneration (AMD), and 20 cases with diabetic retinopathy (DR). Protein levels of sVAP-1, intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, and vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay. Enzymatic activity of semicarbazide-sensitive amine oxidase (SSAO) was also measured. Results: Serum level of sVAP-1 showed a moderate correlation with SSAO activity in all cases. Patients with DR had higher levels of serum sVAP-1 than subjects with ERM and MH, or those with AMD; however, severity of DR is not related to the serum levels of sVAP-1. Serum sVAP-1 correlated positively with VEGF in patients with DR, but not in those with ERM and MH, or those with AMD. Neither soluble ICAM-1 nor VCAM-1 correlated with VEGF, even in subjects with DR. Conclusion: The current data demonstrate the elevated serum levels of sVAP-1 and correlation between sVAP-1 and VEGF in patients with type II diabetes. © 2013 Elsevier Inc. All rights reserved.

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Aquaporin 1-positive stromal niche-like cells directly interact with N-cadherin-positive clusters in the basal limbal epithelium

Higa Kazunari, Kato Naoko, Yoshida Satoru, Ogawa Yoko, Shimazaki Jun, Tsubota Kazuo, Shimmura Shigeto

Stem Cell Research 10 ( 2 ) 147 - 155 2013.03

ISSN 1873-5061

Stem cells have a specialized microenvironment for maintaining self-renewal and multipotent capacities. It is believed that a cornea epithelial stem cell niche exists in the limbus. To characterize the niche of limbal epithelial stem cells, we observed the limbal basal epithelial layer by histological analysis.Cell clusters or cell suspensions from limbal tissue were prepared with collagenase or dispase II and fixed for cytospin sections. Adhesion assays were done to quantitate calcium-dependent cell adhesion. Limbal tissue and cytospin sections were analyzed by immunohistochemistry, transmission electron microscopy and confocal microscopy.AQP1 positive (AQP1⁺) cells were observed as non-epithelial cells in the subepithelial stroma. AQP1 expression did not co-localize with CD31, podoplanin, MART-1 positive cells, but were observed in vimentin positive stromal cells. When we made a thorough search of limbal basal cells by confocal microscopy, AQP1⁺ were observed in the proximity of N-cad, K15 and p63 positive limbal basal epithelial cells. Furthermore, electron microscope revealed stromal cells penetrating the epithelial basal membrane and forming calcium-dependent cellular adhesions with N-cad⁺ limbal basal epithelial cells.Although we could not clearly detect the expression of N-cad in the AQP1⁺ cells, AQP1⁺ cells immediately beneath the epithelial basement membrane may be stromal niche-like cells that directly interact with N-cad⁺ limbal basal epithelial progenitor cells. © 2012 Elsevier B.V.

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Validation of different sets of criteria for the diagnosis of Sjögren's syndrome in Japanese patients

Tsuboi Hiroto, Hagiwara Shinya, Asashima Hiromitsu, Umehara Hisanori, Kawakami Atsushi, Nakamura Hideki, Sano Hajime, Tsubota Kazuo, Ogawa Yoko, Takamura Etsuko, Saito Ichiro, Inoue Hiroko, Nakamura Seiji, Moriyama Masafumi, Takeuchi Tsutomu, Tanaka Yoshiya, Hirata Shintaro, Mimori Tsuneyo, Matsumoto Isao, Sumida Takayuki

Modern Rheumatology 23 ( 2 ) 219 - 225 2013.03

ISSN 1439-7595

Objective: To validate the revised Japanese Ministry of Health criteria for the diagnosis of Sjögren's syndrome (SS) (JPN) (1999), The American-European Consensus Group classification criteria for SS (AECG) (2002), and American College of Rheumatology classification criteria for SS (ACR) (2012). Methods: The study subjects were 694 patients with SS or suspected SS who were followed-up in June 2012 at ten hospitals that form part of the Research Team for Autoimmune Diseases, The Research Program for Intractable Disease by the Ministry of Health, Labor and Welfare (MHLW). All patients had been checked for all four criteria of the JPN (pathology, oral, ocular, anti-SS-A/SS-B antibodies). We studied the clinical diagnosis made by the physician in charge and the satisfaction of the above criteria. Results: Of the 694 patients, 499 patients did not have other connective tissue diseases (CTDs). SS was diagnosed in 476 patients (primary SS in 302, secondary SS in 174), whereas non-SS was diagnosed in 218 patients (without other CTDs in 197, with other CTDs in 21) by the physician in charge. The sensitivities of JPN, AECG, and ACR in the diagnosis of all forms of SS (both primary and secondary SS) were 79.6, 78.6, and 77.5 %, respectively, with respective specificities of 90.4, 90.4, and 83.5 %. The sensitivities of the same systems in the diagnosis of primary SS were 82.1, 83.1, and 79.1 %, respectively, with specificities of 90.9, 90.9, and 84.8 %, respectively. The sensitivities of the same systems in the diagnosis of secondary SS were 75.3, 70.7, and 74.7 %, respectively, with specificities of 85.7, 85.7, and 71.4 %, respectively. Conclusion: The sensitivity of JPN to all forms of SS and secondary SS, the sensitivity of AECG to primary SS, and the specificities of JPN and AECG for all forms of SS, primary SS, and secondary SS were highest in the diagnosis of SS in Japanese patients. These results indicate that the JPN criteria for the diagnosis of SS in Japanese patients are superior to ACR and AECG. © Japan College of Rheumatology 2012.

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Improvements in sleep quality and gait speed after cataract surgery

Ayaki Masahiko, Muramatsu Masahiro, Negishi Kazuno, Tsubota Kazuo

Rejuvenation Research 16 ( 1 ) 35 - 42 2013.02

ISSN 1549-1684

Background: Gait speed and sleep quality are health indices related to longevity and mortality. In the present study, we measured sleep quality, quality of life, gait speed, and visual acuity before and after cataract surgery to evaluate the efficacy of the procedure on systemic health. Methods: The study was conducted on 155 patients (93 women; average age 74.8 years) undergoing cataract surgery with the implantation of a yellow soft acrylic lens. Patients were evaluated using the Pittsburgh Sleep Quality Index (PSQI) and the National Eye Institute Visual Function Questionnaire 25 (VFQ-25; vision-related quality of life) before and then 2 and 7 months after surgery. Four-meter gait speed was also determined. Results: Of the 155 patients, 68 (43.9%) were classified as poor sleepers (PSQI>5.5) prior to surgery. Significant improvements were noted in sleep 2 months after surgery (p<0.05, paired t-test), but thereafter the improvements were not significant. Prior to surgery, 117 patients (77.0%) were classified as slow walkers (speed<1.0 meter/s). Gait speed increased significantly in these patients 2 months after surgery (p<0.001, paired t-test). Multiple regression analysis revealed significant correlations between the preoperative VFQ-25 score and both PSQI (p<0.05) and gait speed (p<0.001). Postoperative increases in the VFQ-25 score were positively correlated with decreases in the PSQI (p<0.05). Improvements in visual acuity were correlated with improvements in the VFQ-25 score, but not with either PSQI or gait speed. Conclusion: Cataract surgery effectively improves sleep quality and slow gait speed. © 2013, Mary Ann Liebert, Inc.

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A case of impaired vision due to glistening and whitening of hydrophobic acrylic intraocular lens

Hidaka Yuki, Negishi Kazuno, Matsushima Hiroyuki, Torii Hidemasa, Saiki Megumi, Watanabe Kazuhiro, Inafuku Saori, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 67 ( 2 ) 199 - 202 2013.02

ISSN 0370-5579

To report a case who received exchange of intraocular lens (IOP) for impaired vision due to glistening and whitening. Case: A 78-year-old woman presented with impaired vision in her right eye. She had received cataract surgery with implantation of IOL 6 years before. She had had reportedly visual acuity of 0.8 immediately after surgery. Findings: Corrected visual acuity was 0.4 in the right eye. Slitlamp examination showed glistening and whitening of IOP. Funduscopy showed slight chorioretinal atrophy which was the same as immediately after surgery. She received exchange of IOL resulting in corrected visual acuity of 0.8. Conclusion: This case illustrates that glistening and whitening of IOL may result in impaired visual function. Exchange of IOL may be useful in such a situation.

Effect of pupil size on uncorrected visual acuity in pseudophakic eyes with astigmatism

Watanabe Kazuhiro, Negishi Kazuno, Dogru Murat, Yamaguchi Takefumi, Torii Hidemasa, Tsubota Kazuo

Journal of Refractive Surgery 29 ( 1 ) 25 - 29 2013.01

ISSN 1081-597X

PURPOSE: To study the effect of pupil size on the relationship between refractive astigmatism and uncorrected distance visual acuity (UDVA) and pseudoaccommodation in pseudophakic eyes. METHODS: Thirty-six eyes from 36 patients who underwent phacoemulsifi cation and intraocular lens (IOL) implantation were included. All eyes were divided into two groups based on the presence of postoperative with-therule astigmatism or against-the-rule astigmatism. Uncorrected and corrected distance visual acuity (CDVA), refractive astigmatism, subjective accommodation amplitude, and pupil sizes under photopic and mesopic conditions were measured 1 month postoperatively. The effects of pupil size on the relationship between refractive astigmatism and UDVA were investigated. RESULTS: Against-the-rule astigmatism had a moderate but signifi cant negative linear correlation with UDVA irrespective of the pupil size (R²=0.60, P<.01). A moderate but signifi cant negative linear correlation was found when the pupil diameter exceeded 2.9 mm in eyes with with-the-rule astigmatism (R²=0.46, P=.04). Refractive astigmatism was not signifi cantly correlated with pseudoaccommodation regardless of the pupil diameter in either group. CONCLUSIONS: The results of the current study suggest that pupil size may have an impact on postoperative UDVA in eyes having against the-rule astigmatism and in eyes with a large pupil diameter and with-the-rule astigmatism. Copyright © SLACK Incorporated.

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Comparison of long-term outcome of penetrating and deep anterior lamellar keratoplasty

Adachi Sayaka, Ichihashi Yoshiyuki, Kawakita Tetsuya, Matsumoto Yukihiro, Uchino Miki, Shimmura Shigeto, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 67 ( 1 ) 85 - 89 2013.01

ISSN 0370-5579

Purpose: To report and compare the long-term outcome of penetrating and deep anterior lamellar keratoplasty. Cases and Method: This retrospective study was made on 36 cases who received keratoplasty by a single surgeon during past 10 years. The series comprised 27 males and 9 females. There were 25 eyes of keratoconus, 7 eyes of corneal leukoma, and 3 eyes of corneal degeneration. Deep anterior lamellar keratoplasty was performed on 21 cases and penetrating keratoplasty on 21 eyes. Cases were followed up for 2 to 10 years, average 5 years. Results: Final visual acuity averaged 0.72 after lamellar and 0.85 after penetrating keratoplasty. There was no significant difference. Spherical equivalent of refraction showed a tendency to myopia with no difference between the two groups. Corneal endothelial cell density decreased by 11.0±6.6% immediately after lamellar and 59.5±29.0% after penetrating keratoplasty (p< 0.01). Conclusion: There was no difference in visual function and safety between deep anterior lamellar keratoplasty and penetrating keratoplasty for 2 years or longer.

Effect of experimentally induced astigmatism on functional, conventional, and low-contrast visual acuity

Watanabe Kazuhiro, Negishi Kazuno, Kawai Miho, Torii Hidemasa, Kaido Minako, Tsubota Kazuo

Journal of Refractive Surgery 29 ( 1 ) 19 - 24 2013.01

ISSN 1081-597X

PURPOSE: To report the changes in functional visual acuity (FVA) in eyes with experimentally induced astigmatism. METHODS: This study included 26 right eyes from 26 healthy participants (mean age: 27.0±4.4 years). After confi rming best correction under cycloplegia, the cylinder power ×90° and ×180° (against-the-rule [ATR] and with-the-rule [WTR] astigmatism, respectively) were added with 0.50-diopter (D) steps, from 0.50 to 2.50 D. Conventional visual acuity (VA), FVA as measured by the FVA Measurement System (NIDEK Co Ltd), and 10% low-contrast visual acuity (LCVA) were measured with 3-mm artifi cial pupil under each condition. RESULTS: The mean corrected logMAR VA, FVA, LCVA were -0.18±0.00 (20/13), -0.10±0.08 (20/16), and 0.06±0.06 (20/23), respectively. The VA, FVA, and LCVA had a signifi cant linear negative correlation with the addition of cylinder irrespective of the axes. A significant decrease in FVA was noted on addition of ≥0.50 D WTR astigmatism and ≥1.00 D ATR astigmatism. With the addition of 1.00 D of cylindrical lens, 96% of eyes with WTR and ATR astigmatism could maintain 20/20 in VA testing, whereas only 50% and 62% of eyes with WTR and ATR astigmatism, respectively, could maintain 20/20 in FVA testing. CONCLUSIONS: Astigmatism may be associated with deterioration of visual function even when conventional VA of 20/20 is attained. Functional VA testing seems to be useful in evaluating the masked astigmatic visual impairment that cannot be detected by conventional VA testing. Copyright © SLACK Incorporated.

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Evaluation of lipid oxidative stress status in Sjögren syndrome patients

Wakamatsu Tais H., Dogru Murat, Matsumoto Yukihiro, Kojima Takashi, Kaido Minako, Ibrahim Osama M A, Sato Enrique A., Igarashi Ayako, Ichihashi Yoshiyuki, Satake Yoshiyuki, Shimazaki Jun, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 54 ( 1 ) 201 - 210 2013.01

ISSN 0146-0404

PURPOSE. We evaluated the levels of lipid oxidative stress markers and inflammatory cells from tears and conjunctiva of patients with Sjögren syndrome (SS) and normal subjects. METHODS. We examined 31 eyes of 16 patients (16 females) with SS and 15 eyes of 10 healthy controls (2 males and 8 females) in this prospective study. All subjects underwent a Schirmer test, measurement of tear film break-up time, vital stainings, confocal microscopy of the conjunctiva, tear collection for hexanoyl-lysine (HEL), ELISA, and conjunctival brush cytology. Brush cytology samples underwent immunohistochemistry (IHC) staining with HEL and 4-hydroxy-2- nonenal (4HNE). Hematoxylin-eosin and IHC staining with HEL and 4HNE also were performed on conjunctival samples of SS patients and controls. RESULTS. The tear stability and vital staining scores were significantly worse in eyes of SS patients compared to the controls. Conjunctival inflammatory cell density was significantly higher in SS subjects compared to controls. The numbers of conjunctival cells stained positively for HEL and 4HNE were significantly higher in SS patients compared to controls. Tear HEL concentrations correlated significantly with staining scores and inflammatory cell density in confocal microscopy. Conjunctival specimens also revealed higher numbers of cells stained positively for inflammatory markers, as well as HEL and 4HNE in the IHC stainings. CONCLUSIONS. Increase of the oxidative stress status in the conjunctiva of SS patients appears to have a role in the pathogenesis of dry eye disease. A close relationship may exist between reactive oxygen species (ROS) production, lipid peroxidation related membrane damage, and inflammatory processes in dry eye. © 2013 The Association for Research in Vision and Ophthalmology, Inc.

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Morphological evaluation of meibomian glands using noncontact infrared meibography

Ban Yumiko, Shimazaki-Den Seika, Tsubota Kazuo, Shimazaki Jun

Ocular Surface 11 ( 1 ) 47 - 53 2013.01

ISSN 1542-0124

Meibography is used to study morphological changes in the meibomian glands (MGs), and semiquantitative analysis has been used for MG assessment. We conducted a detailed morphometric assessment of MGs using noncontact infrared meibography (NIM) and investigated the relationship between MG morphology and the ocular surface and MG morphology associated with aging and sex in 37 subjects. The MGs in the upper and lower eyelids showed significant correlations in their morphology, with the former having longer ducts than the latter. The mean length of the MG ducts, percent area of MG acini, and number of gland dropouts were significantly correlated with age. The mean length of the MG ducts in the upper and lower eyelids showed negative correlations with the meibum (P=.043) and fluorescein staining score (P=.008), respectively. The percent area of MG acini in the upper eyelid showed a positive correlation with tear film breakup time (P=.012) and negative correlations with tear film lipid layer interferometry and meibum (P=.005 and .002, respectively). The mean length of the male's MG ducts in the lower eyelid was significantly longer than that of the females (P=.03). These results indicate that morphometric analysis using NIM may be useful for assessing ocular surface conditions. © 2013 Elsevier Inc.

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Renin-angiotensin system involvement in the oxidative stress-induced neurodegeneration of cultured retinal ganglion cells

Ozawa Yoko, Yuki Kenya, Yamagishi Reiko, Tsubota Kazuo, Aihara Makoto

Japanese Journal of Ophthalmology 57 ( 1 ) 126 - 132 2013.01

ISSN 0021-5155

Purpose: To analyze the influence of oxidative stress on retinal ganglion cells (RGCs) by using a selective culture system of rat RGCs. Methods: Rat RGCs were purified by a two-step immunopanning procedure and cultured either with or without antioxidant (AO) compounds. Reactive oxygen species (ROS) in RGCs were analyzed using dihydroethidium. Expression of angiotensin II, cleaved caspase 3, and netrin-1 was analyzed by immunocytochemistry. Live RGCs were detected by use of calcein-acetoxymethyl ester. The roles of angiotensin II type 1 receptor (AT1R) signaling and netrin-1 were analyzed by use of an AT1R blocker (telmisartan) and an anti-netrin-1 neutralizing antibody, respectively. Results: ROS and angiotensin II were induced in RGCs cultured without AO compounds (AO-). In these cultures, the number of live RGCs decreased and expression of cleaved and activated caspase 3 increased, but these changes were attenuated by addition of the AT1R blocker. Reduction in netrin-1 expression under the AO- condition was also prevented by the AT1R blocker. The AT1R blocker's effects on RGC survival and reduction in cleaved caspase 3-positive cells were cancelled by the anti-netrin-1 neutralizing antibody. Conclusions: Oxidative stress induced cell death through AT1R signaling and netrin-1 reduction in cultured RGCs. © 2012 Japanese Ophthalmological Society.

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[Development of a monitor for quantifying personal eye exposure to visible and ultraviolet radiation and its application in epidemiology].

Eto Norihito, Tsubota Kazuo, Tanaka Taichiro, Nishiwaki Yuji

Nihon eiseigaku zasshi. Japanese journal of hygiene 68 ( 2 ) 118 - 125 2013

ISSN 0021-5082

Eye diseases including cataract, keratitis and pterygium have been reported to be sun-exposure-related. The association between macular degeneration and blue light has also been discussed. Moreover, it is hypothesized that retinal exposure to blue light may influence the human circadian rhythm. However, no monitoring devices exist that can measure eye exposure to visible and ultraviolet (UV) radiation over time. To measure the exact dose at specific times, we have developed a novel sensing system (ray-sensing glass system: RaySeG). RaySeG can continuously measure and record the composition and intensity of light with a time-stamped system. Subjects wearing RaySeG were instructed to walk under various light conditions such as indoor and outdoor. RaySeG consists of two sensors embedded in the eyeglasses. These sensors are for UV (260-400 nm), visible lights (red, 615 nm; green, 540 nm; and blue, 465 nm: peak wavelength for each). The total weight of the system is about 100 g, and the size is comparable to that of a digital audio player. The system continuously recorded changes in visible and UV light exposure under various conditions. After accuracy validation, further experiments with a larger number of subjects are required. Our final goal is to apply the system to evaluating personal eye exposure to UV and visible light in epidemiological studies of eye diseases and circadian rhythm abnormality.

Expression and localization of aging markers in lacrimal gland of chronic graft-versus-host disease.

Kawai Masataka, Ogawa Yoko, Shimmura Shigeto, Ohta Shigeki, Suzuki Takanori, Kawamura Naoshi, Kuwana Masataka, Kawakami Yutaka, Tsubota Kazuo

Scientific Reports 3 2455 2013

Aging is commonly defined as the accumulation of diverse deleterious changes in cells and tissues with advancing age. To investigate whether aging changes are involved in the lacrimal glands of chronic graft-versus-host disease (cGVHD) model mice, we obtained the specimens from cGVHD model mice, untreated aged and young mice, and examined by histopathology, and immunoblotting. Oxidative stress markers, 8-OHdG, 4-HNE, and hexonoyl lesion (HEL), and other aging markers, p16 and p38, were used to assess the samples. The infiltrating mononuclear cells and endothelia of capillaries in the cGVHD and aged mice expressed the oxidative stress markers and other aging markers, but not in the young mice. Histological changes and the expression of aging markers in the samples from cGVHD mice exhibited similar features to those in aging mice. These results suggest that changes that typically appear with advanced age occur earlier in the lives of mice with lacrimal gland cGVHD.

Neuroprotective role of VEGF induced in response to photodynamic therapy

Ozawa Yoko, Suzuki Misa, Niikura Serina, Kamoshita Mamoru, Nagai Norihiro, Tsubota Kazuo

Advances in Neuroimmune Biology 4 ( 3 ) 161 - 165 2013

ISSN 1878-948X

Age-related macular degeneration (AMD) is a vision-threatening disease, and its prevalence is increasing worldwide. Two approaches are commonly used to suppress the pathological choroidal neovascularization (CNV) that characterizes 'wet' AMD. One is photodynamic therapy (PDT), which involves using a systemic photosensitizer in combination with low-power laser irradiation of CNV through the neural retina and retinal pigment epithelium (RPE); the other involves the intravitreous injection of drugs targeting vascular endothelial growth factor (VEGF). In some cases, both methods are used simultaneously. However, recent results from animal experiments indicate that transient increase in VEGF in the tissue following PDT is required for retinal cell survival in overcoming the cellular stress caused by PDT. To maximize the visual outcome and retinal protection, it will be necessary to optimize the combination therapy, which requires the elucidation of the molecular and cellular changes that occur in the retinal tissue during the course of treatment. © 2013 - IOS Press and the authors. All rights reserved.

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ABCA4 gene screening by next-generation sequencing in a British cohort

Fujinami Kaoru, Zernant Jana, Chana Ravinder K., Wright Genevieve A., Tsunoda Kazushige, Ozawa Yoko, Tsubota Kazuo, Webster Andrew R., Moore Anthony T., Allikmets Rando, Michaelides Michel

Investigative Ophthalmology and Visual Science 54 ( 10 ) 6662 - 6674 2013

ISSN 0146-0404

Purpose. We applied a recently reported next-generation sequencing (NGS) strategy for screening the ABCA4 gene in a British cohort with ABCA4-associated disease and report novel mutations. Methods. We identified 79 patients with a clinical diagnosis of ABCA4-associated disease who had a single variant identified by the ABCA4 microarray. Comprehensive phenotypic data were obtained, and the NGS strategy was applied to identify the second allele by means of sequencing the entire coding region and adjacent intronic sequences of the ABCA4 gene. Identified variants were confirmed by Sanger sequencing and assessed for pathogenicity by in silico analysis. Results. Of the 42 variants detected by prescreening with the microarray, in silico analysis suggested that 34, found in 66 subjects, were disease-causing and 8, found in 13 subjects, were benign variants. We detected 42 variants by NGS, of which 39 were classified as disease-causing. Of these 39 variants, 31 were novel, including 16 missense, 7 splice-site-altering, 4 nonsense, 1 in-frame deletion, and 3 frameshift variants. Two or more disease-causing variants were confirmed in 37 (47%) of 79 patients, one disease-causing variant in 36 (46%) subjects, and no disease-causing variant in 6 (7%) individuals. Conclusions. Application of the NGS platform for ABCA4 screening enabled detection of the second disease-associated allele in approximately half of the patients in a British cohort where one mutation had been detected with the arrayed primer extension (APEX) array. The time- and cost-efficient NGS strategy is useful in screening large cohorts, which will be increasingly valuable with the advent of ABCA4-directed therapies. © 2013 The Association for Research in Vision and Ophthalmology, Inc.

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Functional lacrimal gland regeneration by transplantation of a bioengineered organ germ

Hirayama Masatoshi, Ogawa Miho, Oshima Masamitsu, Sekine Yurie, Ishida Kentaro, Yamashita Kentaro, Ikeda Kazutaka, Shimmura Shigeto, Kawakita Tetsuya, Tsubota Kazuo, Tsuji Takashi

Nature Communications 4 2013

The lacrimal gland has a multifaceted role in maintaining a homeostatic microenvironment for a healthy ocular surface via tear secretion. Dry-eye disease, which is caused by lacrimal gland dysfunction, is one of the most prevalent eye diseases that cause corneal epithelial damage and results in significant loss of vision and a reduction in the quality of life. Here we demonstrate orthotopic transplantation of bioengineered lacrimal gland germs into adult mice with an extra-orbital lacrimal gland defect, a mouse model that mimics the corneal epithelial damage caused by lacrimal gland dysfunction. The bioengineered lacrimal gland germs and harderian gland germs both develop in vivo and achieve sufficient physiological functionality, including tear production in response to nervous stimulation and ocular surface protection. This study demonstrates the potential for bioengineered organ replacement to functionally restore the lacrimal gland. © 2013 Macmillan Publishers Limited. All rights reserved.

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Tear lipid layer deficiency associated with incomplete blinking

Kawashima Motoko, Tsubota Kazuo

BMC Ophthalmology 13 ( 1 ) 2013

Abstract. Background: Meibomian gland obstruction induces hyposecretion of tear film lipids, which results in lipid layer deficiency and evaporative dry eye. Unfortunately, the importance of blinking in meibomian gland dysfunction has been largely overlooked, and it is not known whether incomplete blinking causes tear lipid deficiency, even in the unobstructed meibomian glands. Case presentation. A 38-year-old woman suffering from foreign body sensations in her eyes was examined. The cornea was clear and tear secretion was normal. Lid margin abnormalities were not observed and the meibum was clear. However, the lipid layer was very thin, and the patient was given a diagnosis of incomplete blinking. The patient was made aware of her condition and asked to blink consciously and completely. After that, an immediate increase in lipid flow was observed. Conclusion: Tear lipid layer deficiency can occur with incomplete blinking, even though meibomian gland structures are intact. This case highlights the importance of complete blinking. © 2013 Kawashima and Tsubota; licensee BioMed Central Ltd.

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Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithelium of light-exposed mice

Narimatsu Toshio, Ozawa Yoko, Miyake Seiji, Kubota Shunsuke, Hirasawa Manabu, Nagai Norihiro, Shimmura Shigeto, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 54 ( 7 ) 4555 - 4562 2013

ISSN 0146-0404

PURPOSE. To elucidate the influences of light exposure on the retinal pigment epithelium (RPE) in vivo that may be involved in the pathogenesis of AMD. METHODS. Six- to 7-week-old BALB/c mice were exposed to light at 2000 lux for 3 hours. Flatmount RPE samples were immunostained with anti-ZO-1 antibody for evaluating tight junction, anti-N-cadherin, and anti-b-catenin antibodies for adherens junction, and stained with phalloidin for actin cytoskeleton. The reactive oxygen species (ROS) level was measured using DCFH-DA; Rho-associated coiled-coil forming kinase (ROCK) activity was by ELISA. Cytokine expression was analyzed by real-time RT-PCR and/or ELISA in the RPE-choroid, and macrophage recruitment was by real-time RT-PCR and immunohistochemistry. Either an antioxidant, N-Acetyl-L-cysteine (NAC), or a ROCK inhibitor, Y-27632, were administered to analyze the roles of ROS and ROCK activation, respectively. RESULTS. Light exposure disrupted staining patterns of tight junctions, adherens junctions, and actin cytoskeleton in the RPE, where ROS was elevated. However, NAC treatment avoided the RPE changes, reducing ROS. ROCK activity increased after light exposure was suppressed by NAC, and the structural disruptions were suppressed by Y-27632. The levels of MCP-1, CCL11, and IL-6 increased after light exposure were suppressed by NAC. Light-induced MCP-1 and IL- 6 were suppressed by Y-27632. Macrophage recruitment after light exposure was also suppressed either by NAC or Y-27632. CONCLUSIONS. Light exposure induced ROS and Rho/ROCK activation, which caused disruption of cell-cell junctions (tight junctions and adherens junctions) and actin cytoskeleton, the RPE's barrier structure, and induced AMD-associated pathological changes in the RPE-choroid. Copyright 2013 The Association for Research in Vision and Ophthalmology, Inc.

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Effects of dietary supplementation with fish oil on dry eye syndrome subjects

Kawakita Tetsuya, Kawabata Fuminori, Tsuji Tomoko, Kawashima Motoko, Shimmura Shigeto, Tsubota Kazuo

Biomedical Research (Japan) 34 ( 5 ) 215 - 220 2013

ISSN 0388-6107

The purpose of this study was to evaluate the efficacy of fish oil supplementation added to usual dry eye treatment in dry eye subjects in a randomized controlled trial. Twenty-seven typical dry eye subjects were selected from 43 candidates by the diagnostic criterion for dry eye in this study. They were assigned to the randomized fish oil group (n = 15) or the placebo group (n = 12). Fish oil group ingested fish oil capsules containing eicosapentaenoic acid (EPA, 1245 mg/day) and docosahexaenoic acid (DHA, 540 mg/day) for 12 weeks. Placebo group ingested placebo capsules without EPA or DHA. A visual analog scale test estimating subjective symptoms, the Schirmer I test, tear film break-up time (BUT) measurement, fluorescein staining, and rose bengal staining were performed every 4 weeks during the 12-week supplementation period and 4-week washout period. The subjective symptom of "eye pain", BUT, and changes in rose bengal staining score of the fish oil group were significantly improved after 8-12 weeks of supplementation and/or 4 weeks of washout, compared to those of the placebo group. These results suggest that fish oil supplementation added to usual care may be effective in the treatment of dry eye.

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The effect of 3D visual simulator on children's visual acuity - A pilot study comparing two different modalities

Ide Takeshi, Ishikawa Mariko, Tsubota Kazuo, Miyao Masaru

Open Ophthalmology Journal 7 69 - 78 2013

Purpose: To evaluate the efficacy of two non-surgical interventions of vision improvement in children. Methods: A prospective, randomized, pilot study to compare fogging method and the use of head mounted 3D display. Subjects were children, between 5 to 15 years old, with normal best corrected visual acuity (BCVA) and up to -3D myopia. Subjects played a video game as near point work, and received one of the two methods of treatments. Measurements of uncorrected far visual acuity (UCVA), refraction with autorefractometer, and subjective accommodative amplitude were taken 3 times, at the baseline, after the near work, and after the treatment. Results: Both methods applied after near work, improved UCVA. Head mounted 3D display group showed significant improvement in UCVA and resulted in better UCVA than baseline. Fogging group showed improvement in subjective accommodative amplitude. While 3D display group did not show change in the refraction, fogging group's myopic refraction showed significant increase indicating the eyes showed myopic change of eyes after near work and treatment. Discussion: Despite our lack of clear knowledge in the mechanisms, both methods improved UCVA after the treatments. The improvement in UCVA was not correlated to measured refraction values. Conclusion: UCVA after near work can be improved by repeating near and distant accommodation by fogging and 3D image viewing, although at the different degrees. Further investigation on mechanisms of improvements and their clinical significance are warranted. © Ide et al.

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Oral voriconazole induced visual disturbances that led to the diagnosis of optic radiation lesions in a patient with aplastic anemia

Horino Takeshi, Uchida Atsuro, Mochimaru Hiroshi, Koto Takashi, Nagai Norihiro, Shinoda Hajime, Shinoda Kei, Ohde Hisao, Ozawa Yoko, Tsubota Kazuo

Folia Japonica de Ophthalmologica Clinica 5 ( 12 ) 1129 - 1133 2012.12

ISSN 1882-5176

The Semaphorin 3A Inhibitor SM-345431 Accelerates Peripheral Nerve Regeneration and Sensitivity in a Murine Corneal Transplantation Model

Omoto Masahiro, Yoshida Satoru, Miyashita Hideyuki, Kawakita Tetsuya, Yoshida Kenji, Kishino Akiyoshi, Kimura Toru, Shibata Shinsuke, Tsubota Kazuo, Okano Hideyuki, Shimmura Shigeto

PLoS ONE 7 ( 11 ) 2012.11

Background: Peripheral nerve damage of the cornea is a complication following surgery or infection which may lead to decreased visual function. We examined the efficacy of the semaphorin 3A inhibitor, SM-345431, in promoting regeneration of peripheral nerves in a mouse corneal transplantation model. Methodology/Principal Findings: P0-Cre/Floxed-EGFP mice which express EGFP in peripheral nerves cells were used as recipients of corneal transplantation with syngeneic wild-type mouse cornea donors. SM-345431 was administered subconjunctivally every 2 days while control mice received vehicle only. Mice were followed for 3 weeks and the length of regenerating nerves was measured by EGFP fluorescence and immunohistochemistry against βIII tubulin. Cornea sensitivity was also measured by the Cochet-Bonnet esthesiometer. CD31 staining was used to determine corneal neovascularization as a possible side effect of SM-345431. Regeneration of βIII tubulin positive peripheral nerves was significantly higher in SM-345431 treated mice compared to control. Furthermore, corneal sensitivity significantly improved in the SM-345431 group by 3 weeks after transplantation. Neovascularization was limited to the peripheral cornea with no difference between SM-345431 group and control. Conclusions/Significance: Subconjunctival injections of SM-345431 promoted a robust network of regenerating nerves as well as functional recovery of corneal sensation in a mouse keratoplasty model, suggesting a novel therapeutic strategy for treating neurotrophic corneal disease. © 2012 Omoto et al.

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Surgical management of lacrimal punctal cauterization in chronic GVHD-related dry eye with recurrent punctal plug extrusion

Yaguchi S., Ogawa Y., Kamoi M., Uchino M., Tatematsu Y., Ban Y., Ohba E., Okamoto S., Goto E., Tsubota K.

Bone Marrow Transplantation 47 ( 11 ) 1465 - 1469 2012.11

ISSN 0268-3369

We investigated the efficacy of lacrimal punctal occlusion surgery with a cautery device in patients with chronic GVHD (cGVHD)-related dry eye, with recanalization of puncta and recurrent punctal plug extrusion. A total of 23 puncta from 14 eyes of 10 patients with chronic GVHD (cGVHD)-related dry eye underwent punctual thermal cauterization with a high-temperature disposable cautery device. All patients were refractory to conventional treatment, including artificial tear eye drops, autologous serum eye drops and vitamin A eye drops, and had a history of recanalization and recurrent punctal plug extrusion. The effect of lacrimal punctal cauterization by thermal cautery device was evaluated by changes in subjective symptom scores, corrected distance visual acuity, Schirmer's test values, fluorescein staining scores, rose bengal staining scores, and tear-film break-up time before and 3 months after the surgery. Subjective symptom scores, Schirmer's test values, fluorescein and rose bengal scores, and tear-film break-up time improved significantly 3 months after the surgery. Recanalization of puncta was not observed in all the cases (0 of 14 eyes, 0%). Lacrimal punctal cauterization was effective with no recanalization and significant improvements in subjective symptoms and the ocular surface environment in cGVHD-related dry eye patients who had been refractory to conventional treatments. © 2012 Macmillan Publishers Limited All rights reserved.

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The antiaging approach for the treatment of dry eye

Tsubota Kazuo, Kawashima Motoko, Inaba Takaaki, Dogru Murat, Matsumoto Yukihiro, Ishida Reiko, Kaido Minako, Kojima Takashi, Uchino Miki, Uchino Yuichi, Ogawa Yoko, Nakamura Shigeru, Higuchi Akihiro, Shinmura Ken, Watanabe Mitsuhiro, Kawakita Tetsuya

Cornea 31 ( 11 SUPPL.1 ) 2012.11

ISSN 0277-3740

ABSTRACT:: Dry eye is one of the most common eye disorders affecting millions of people. It causes ocular irritation or discomfort, and decreases functional vision, causing a dramatic deterioration in the quality of life. Although new treatments such as the P2Y2 agonist or cyclosporine eye drops have been developed and a certain level of patient satisfaction can now be obtained, no fundamental treatment has been developed. Currently, there is no therapy available to recover lacrimal function to its normal status. Recent progress in the understanding of aging has laid the foundations for a new way of thinking about intervention of the aging process. Because dry eye is accelerated by aging, a useful approach for the prevention or treatment of dry eye may be to interfere with the aging process. In the scientific community, there is a global consensus that calorie restriction can extend the life span of various kinds of animals, establishing an intervention to aging. Another important hypothesis believed to be involved in aging is the free radical theory. According to these theories, the aging process may be managed by controlling levels of calories or reactive oxygen species. In this review, these 2 important aging theories, calorie restriction and free radical aging, are examined, and we discuss how to apply these theories to the prevention and treatment of dry eye. Copyright © 2012 by Lippincott Williams & Wilkins.

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A new mouse model of dry eye disease

Uchino Yuichi, Kawakita Tetsuya, Ishii Takamasa, Ishii Naoaki, Tsubota Kazuo

Cornea 31 ( 11 SUPPL.1 ) 2012.11

ISSN 0277-3740

PURPOSE:: Oxidative damage and inflammation are proposed to be involved in the age-related functional decline of lacrimal glands. The molecular mechanism(s) of how oxidative stress affects the secretory function of lacrimal glands was investigated because this is currently unclear. METHODS:: We used a novel mev-1 conditional transgenic mouse model (Tet-mev-1) with a modified tetracycline system. The mev-1 gene encodes the cytochrome b560 large subunit of succinate-ubiquinone oxidoreductase in complex II of mitochondria. RESULTS:: Expression of the mev-1 gene induced excessive oxidative stress associated with ocular surface epithelial damage and a decrease in aqueous secretory function. Tear volume in Tet-mev-1 mice was lower than in wild-type mice, and histopathological analyses showed the hallmarks of lacrimal gland inflammation by intense mononuclear leukocytic infiltration and fibrosis in the lacrimal gland of Tet-mev-1 mice. CONCLUSIONS:: This new model provides evidence that mitochondria-induced oxidative damage in the lacrimal gland induces lacrimal dysfunction, resulting in dry eye disease. Our findings strongly suggest that oxidative stress can be a causative factor in the development of dry eye disease. Copyright © 2012 by Lippincott Williams & Wilkins.

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Oxidative Stress Induced Inflammation Initiates Functional Decline of Tear Production

Uchino Yuichi, Kawakita Tetsuya, Miyazawa Masaki, Ishii Takamasa, Onouchi Hiromi, Yasuda Kayo, Ogawa Yoko, Shimmura Shigeto, Ishii Naoaki, Tsubota Kazuo

PLoS ONE 7 ( 10 ) 2012.10

Oxidative damage and inflammation are proposed to be involved in an age-related functional decline of exocrine glands. However, the molecular mechanism of how oxidative stress affects the secretory function of exocrine glands is unclear. We developed a novel mev-1 conditional transgenic mouse model (Tet-mev-1) using a modified tetracycline system (Tet-On/Off system). This mouse model demonstrated decreased tear production with morphological changes including leukocytic infiltration and fibrosis. We found that the mev-1 gene encodes Cyt-1, which is the cytochrome b₅₆₀ large subunit of succinate-ubiquinone oxidoreductase in complex II of mitochondria (homologous to succinate dehydrogenase C subunit (SDHC) in humans). The mev-1 gene induced excessive oxidative stress associated with ocular surface epithelial damage and a decrease in protein and aqueous secretory function. This new model provides evidence that mitochondrial oxidative damage in the lacrimal gland induces lacrimal dysfunction resulting in dry eye disease. Tear volume in Tet-mev-1 mice was lower than in wild type mice and histopathological analyses showed the hallmarks of lacrimal gland inflammation by intense mononuclear leukocytic infiltration and fibrosis in the lacrimal gland of Tet-mev-1 mice. These findings strongly suggest that oxidative stress can be a causative factor for the development of dry eye disease. © 2012 Uchino et al.

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Efficacy and safety of diquafosol ophthalmic solution in patients with dry eye syndrome

Matsumoto Yukihiro, Ohashi Yuichi, Watanabe Hitoshi, Tsubota Kazuo

Ophthalmology 119 ( 10 ) 1954 - 1960 2012.10

ISSN 0161-6420

Objective: To investigate the dose-dependent efficacy and safety of diquafosol ophthalmic solution for the treatment of dry eye syndrome. Design: Randomized, double-masked, multicenter, parallel-group, placebo-controlled trial. Participants: A total of 286 Japanese patients with dry eye who were prescribed topical diquafosol (1%, n = 96; 3%, n = 96) or placebo ophthalmic solution (n = 94). Methods: After a washout period of 2 weeks, qualified subjects were randomized to receive a single drop of 1% or 3% diquafosol or placebo ophthalmic solutions 6 times per day for 6 weeks. Main Outcome Measures: The primary outcome measure was fluorescein corneal staining score assessment. The secondary outcome measures were Rose Bengal corneal and conjunctival staining scores, tear break-up time (BUT), and subjective symptom assessment. Safety measures were clinical blood and urine examination and recording of adverse events. Results: Fluorescein corneal staining scores significantly improved with both 1% and 3% topical diquafosol compared with placebo at 4 weeks, respectively (P = 0.037, P = 0.002). There was a dose-dependent effect among the groups. Rose Bengal corneal and conjunctival staining scores also improved significantly with both 1% and 3% diquafosol compared with placebo (P = 0.007 and P = 0.004, respectively). Subjective dry eye symptom scores significantly improved with both diquafosol ophthalmic solutions (P ≤ 0.033), although there were no significant differences in BUT compared with placebo. No significant differences between the treatment groups were observed in relation to the occurrence of adverse events. Conclusions: Both 1% and 3% diquafosol ophthalmic solutions are considered effective and safe for the treatment of dry eye syndrome. Financial Disclosure(s): Proprietary or commercial disclosure may be found after the references. © 2012 American Academy of Ophthalmology.

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In vivo confocal microscopy evaluation of meibomian gland dysfunction in atopic-keratoconjunctivitis patients

Ibrahim Osama M A, Matsumoto Yukihiro, Dogru Murat, Adan Enrique Sato, Wakamatsu Tais Hitomi, Shimazaki Jun, Fujishima Hiroshi, Tsubota Kazuo

Ophthalmology 119 ( 10 ) 1961 - 1968 2012.10

ISSN 0161-6420

Purpose: To clarify meibomian gland (MG) alterations in atopic keratoconjunctivitis (AKC) patients and compare the findings with obstructive MG dysfunction (MGD) patients and control subjects using in vivo confocal microscopy (CM). Design: Prospective, controlled, single-center study. Participants: Twelve AKC patients (10 males, 2 females; mean age, 31.0±16.5 years), 12 obstructive MGD patients (7 males, 5 females; mean age, 37.6±5.6 years), and 26 control subjects (13 males, 13 females; mean age, 32.9±5.7 years) were recruited. No significant age or gender differences were observed between the 3 groups. Methods: All subjects underwent assessment of tear evaporation rate from the ocular surface (TEROS), slit-lamp examinations, tear break-up time (BUT) measurements, vital staining, Schirmer test I, meibography, MG expressibility, and CM examination of the MG (HRTII-RCM). Statistical analysis was performed using the Mann-Whitney test. Main Outcome Measures: The MG acinar unit density, inflammatory cell density, MG acinar unit longest diameter, MG acinar unit shortest diameter, and MG acinar unit area as observed by in vivo CM, MG drop-out, MG expressibility grading, tear stability, tear evaporation, and vital staining scores. Results: The TEROS values, mean BUT, vital staining scores, MG expressibility, and MG dropout grades were significantly worse in AKC patients compared with those in obstructive MGD patients and controls (P<0.05). The mean values of the CM parameters in AKC patients were significantly worse than those observed in the obstructive MGD patients and controls (P<0.001). Conclusions: Changes in MG in AKC patients seem to be more severe than in patients with obstructive MGD and controls. In vivo CM is a noninvasive, efficient tool in the assessment of MG status and ocular surface disease in AKC. Financial Disclosure(s): The authors have no proprietary or commercial interest in any of the materials discussed in this article. © 2012 American Academy of Ophthalmology.

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A randomised, double-masked comparison study of diquafosol versus sodium hyaluronate ophthalmic solutions in dry eye patients

Takamura Etsuko, Tsubota Kazuo, Watanabe Hitoshi, Ohashi Yuichi

British Journal of Ophthalmology 96 ( 10 ) 1310 - 1315 2012.10

ISSN 0007-1161

Aims: To compare the efficacy and safety of 3% diquafosol ophthalmic solution with those of 0.1% sodium hyaluronate ophthalmic solution in dry eye patients, using mean changes in fluorescein and rose bengal staining scores as endpoints. Trial design and methods: In this multicenter, randomised, double-masked, parallel study of 286 dry eye patients with fluorescein and rose bengal staining scores of ≥3 were randomised to the treatment groups in a 1:1 ratio. Efficacy and safety were evaluated after drop-wise instillation of the study drug, six times daily for 4 weeks. Results: After 4 weeks, the intergroup difference in the mean change from baseline in fluorescein staining score was -0.03; this verified the non-inferiority of diquafosol. The mean change from baseline in rose bengal staining score was significantly lower in the diquafosol group (p=0.010), thus verifying its superiority. The incidence of adverse events was 26.4% and 18.9% in the diquafosol and sodium hyaluronate groups, respectively, with no significant difference. Conclusions: Diquafosol (3%) and sodium hyaluronate (0.1%) exhibit similar efficacy in improving fluorescein staining scores of dry eye patients, whereas, diquafosol exhibits superior efficacy in improving rose bengal staining scores. Diquafosol has high clinical efficacy and is well tolerated with a good safety profile.

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Selenium Compound Protects Corneal Epithelium against Oxidative Stress

Higuchi Akihiro, Inoue Hiroyoshi, Kawakita Tetsuya, Ogishima Tadashi, Tsubota Kazuo

PLoS ONE 7 ( 9 ) 2012.09

The ocular surface is strongly affected by oxidative stress, and anti-oxidative systems are maintained in corneal epithelial cells and tear fluid. Dry eye is recognized as an oxidative stress-induced disease. Selenium compound eye drops are expected to be a candidate for the treatment of dry eye. We estimated the efficacy of several selenium compounds in the treatment of dry eye using a dry eye rat model. All of the studied selenium compounds were uptaken into corneal epithelial cells in vitro. However, when the selenium compounds were administered as eye drops in the dry eye rat model, most of the selenium compounds did not show effectiveness except for Se-lactoferrin. Se-lactoferrin is a lactoferrin that we prepared that binds selenium instead of iron. Se-lactoferrin eye drops suppressed the up-regulated expression of heme oxygenase-1, cyclooxygenase-2, matrix metallopeptidase-9, and interleukin-6 and also suppressed 8-OHdG production in the cornea induced by surgical removal of the lacrimal glands. Compared with Se-lactoferrin, apolactoferrin eye drops weakly improved dry eye in high dose. The effect of Se-lactoferrin eye drops on dry eye is possibly due to the effect of selenium and also the effect of apolactoferrin. Se-lactoferrin is a candidate for the treatment of dry eye via regulation of oxidative stress in the corneal epithelium. © 2012 Higuchi et al.

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Accumulation of Secretory Vesicles in the Lacrimal Gland Epithelia Is Related to Non-Sjögren's Type Dry Eye in Visual Display Terminal Users

Kamoi Mizuka, Ogawa Yoko, Nakamura Shigeru, Dogru Murat, Nagai Toshihiro, Obata Hiroto, Ito Masataka, Kaido Minako, Kawakita Tetsuya, Okada Yasunori, Kawakami Yutaka, Shimmura Shigeto, Tsubota Kazuo

PLoS ONE 7 ( 9 ) 2012.09

Previous observations in a rat model of a non-Sjögren's syndrome (non-SS) type of dry eye seen in users of visual display terminals (VDT) indicated that secretory vesicle (SV) accumulation in the lacrimal gland epithelia contributes to the condition. Here, to examine this possibility in humans, we compared the lacrimal gland histology and percent SV area in the cytoplasm of acinar epithelial cells using light microscopy and transmission electron microscopy, in patients with VDT work-related non-SS dry-eye (VDT group), SS-induced dry-eye, and autopsied normal controls. In addition, the VAMP8 (vesicle-associated membrane protein 8, an exocrine-pathway molecule) and Rab3D (mature vesicle marker) were histochemically examined in lacrimal gland tissue sections. The lacrimal gland acini were larger in the VDT group than in the SS group, and the percent SV area was significantly higher in the VDT group than in the normal controls (P = 0.021) or SS group (P = 0.004). Immunostaining revealed abnormal distributions of VAMP8 in the VDT and SS groups. Rab3D was more strongly expressed in the cytoplasm of acinar epithelial cells in the VDT group than in that of normal controls. The duration of VDT use was significantly longer in the VDT group than in the other groups. These findings suggest that excessive SV accumulation in the acinar epithelia may contribute to the reduced tear secretion in VDT users. © 2012 Kamoi et al.

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Comparison of domestic and overseas donor corneas for keratoplasty

Nishi Yasuyo, Kawakita Tetsuya, Yasu Masako, Ichihashi Yoshiyuki, Matsumoto Yukihiro, Shimmura Shigeto, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 66 ( 9 ) 1337 - 1342 2012.09

ISSN 0370-5579

Purpose: To compare the safety of domestic and overseas donor corneas for keratoplasty. Objects and Method: This study was made on 490 eyes donated to us through eyebanks in the past 5 years through 2010. The series comprised 245 eyes from domestic donors and 245 eyes from USA. Results: Age of domestic donors averaged 68.8 years and that of overseas donors averaged 63.1 years. The difference was significant (p<0.01). Corneal endothelial density averaged 2713.3 and 2515.3/mm ² respectively. The difference was significant (p<0.01). Primary graft failure occurred in 4 domestic eyes and none in overseas eyes. There was no difference in the positive rate of culture from corneal limbus or preservation medium between domestic and overseas donor eyes. Conclusion: The mean corneal endothelial density was higher in domestic than in overseas cornea. Primary graft failure was more frequent in domestic than overseas eyes. More strict quality control is desired for donated corneas.

Visual function changes after punctal occlusion with the treatment of short but type of dry eye

Kaido Minako, Ishida Reiko, Dogru Murat, Tsubota Kazuo

Cornea 31 ( 9 ) 1009 - 1013 2012.09

ISSN 0277-3740

Purpose: To evaluate visual function changes after punctal plug (PP) occlusion in patients with short breakup time of tear film (BUT) type of dry eyes. Methods: A prospective comparative study was performed in 43 eyes of 43 dry eye patients with short BUTbreakup time of tear film receiving PP occlusion. Patients were divided into 2 groups as those receiving only upper or only lower occlusion. Functional visual acuity measurements, tear function evaluations, and questionnaire on satisfaction with treatment in relation to presence of epiphora were performed before and 1 month after PP insertion. Results: Twenty eyes of 43 eyes (46.5%) with only lower occlusion and 17 eyes of 43 eyes (39.5%) with only upper occlusion showed satisfactory outcomes with treatment. One of 43 eyes (2.3 %) had epiphora. Vital staining scores and BUT values significantly improved in both groups after occlusion. The mean logarithm of the minimum angle of resolution functional visual acuity value significantly improved only in eyes receiving upper PP occlusion. Conclusion: Upper PP occlusion seems to be associated not only with improvement of tear stability, vital staining scores, and treatment-related satisfaction but also with better quality of visual function. Copyright © 2012 by Lippincott Williams & Wilkins.

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Transplantation of side population cells restores the function of damaged exocrine glands through clusterin

Mishima Kenji, Inoue Hiroko, Nishiyama Tatsuaki, Mabuchi Yo, Amano Yusuke, Ide Fumio, Matsui Makoto, Yamada Hiroyuki, Yamamoto Gou, Tanaka Junichi, Yasuhara Rika, Sakurai Takashi, Lee Masaichi Chang Il, Chiba Kan, Sumimoto Hidetoshi, Kawakami Yutaka, Matsuzaki Yumi, Tsubota Kazuo, Saito Ichiro

Stem Cells 30 ( 9 ) 1925 - 1937 2012.09

ISSN 1066-5099

Stem cell-based therapy has been proposed as a promising strategy for regenerating tissues lost through incurable diseases. Side population (SP) cells have been identified as putative stem cells in various organs. To examine therapeutic potential of SP cells in hypofunction of exocrine glands, SP cells isolated from mouse exocrine glands, namely, lacrimal and salivary glands, were transplanted into mice with irradiation-induced hypofunction of the respective glands. The secretions from both glands in the recipient mice were restored within 2 months of transplantation, although the transplanted cells were only sparsely distributed and produced no outgrowths. Consistent with this, most SP cells were shown to be CD31-positive endothelial-like cells. In addition, we clarified that endothelial cell-derived clusterin, a secretory protein, was an essential factor for SP cell-mediated recovery of the hypofunctioning glands because SP cells isolated from salivary glands of clusterin-deficient mice had no therapeutic potential, whereas lentiviral transduction of clusterin restored the hypofunction. In vitro and in vivo studies showed that clusterin had an ability to directly inhibit oxidative stress and oxidative stress-induced cell damage. Thus, endothelial cell-derived clusterin possibly inhibit oxidative stress-induced hypofunction of these glands. © AlphaMed Press.

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The relation between visual performance and clinical ocular manifestations in Stevens-Johnson syndrome

Kaido Minako, Yamada Masakazu, Sotozono Chie, Kinoshita Shigeru, Shimazaki Jun, Tagawa Yoshitsugu, Hara Yuko, Chikama Taiichiro, Tsubota Kazuo

American Journal of Ophthalmology 154 ( 3 ) 2012.09

ISSN 0002-9394

Purpose: To investigate the relation between visual function, clinical findings, and visual symptoms in Stevens-Johnson syndrome (SJS) and to compare the results with Sjögren syndrome (SS) patients and normal subjects. Design: Cross-sectional comparative study. Methods: One hundred fifteen eyes of 59 consecutive patients with SJS and toxic epidermal necrolysis (TEN), 208 eyes of 104 healthy normal subjects, and 132 eyes of 66 SS patients were investigated in this multicenter study. All study subjects underwent tear function and ocular surface examinations, Landolt and functional visual acuity examinations, and the Japanese version of the NEI VFQ-25 (National Eye Institute Visual Function Questionnaire). Results: The mean ocular surface grading scores were significantly higher and the mean score of all 12 NEI VFQ subscales was significantly lower in the SJS patients compared to the SS patients and the normal subjects (P <.05). The conventional and functional logarithm of minimal angle of resolution (logMAR) visual acuities in SJS patients with minimal corneal complications were significantly higher and the mean total composite NEI VFQ scores were lower compared to SS patients. The conventional and functional logMAR visual acuities and the mean ocular surface grading scores in SJS with aqueous deficiency were significantly higher and the mean total composite NEI VFQ scores were lower compared to SS patients. Strong correlations between best-corrected logMAR functional visual acuities and either ocular surface grading scores or the composite NEI VFQ-25 scores were observed. Conclusions: The functional visual acuity examination reflects the severity of clinical ocular surface findings and vision-related quality of life more than the standard conventional visual acuity in SJS. © 2012 Elsevier Inc. All rights reserved.

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Presence and physiologic function of the renin-angiotensin system in mouse lacrimal gland

Yaguchi Saori, Ogawa Yoko, Shimmura Shigeto, Hatou Shin, Nakamura Shigeru, Inaba Takaaki, Imada Toshihiro, Ozawa Yoko, Kawakami Yutaka, Ishida Susumu, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 53 ( 9 ) 5416 - 5425 2012.08

ISSN 0146-0404

PURPOSE. To investigate the expression, localization, and physiologic function of renin-angiotensin system (RAS) components in the mouse lacrimal gland. METHODS. Lacrimal glands and cultured lacrimal gland fibroblasts from wild-type (WT) BALB/c (H-2^d) mice were used. Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used to determine the expression and localization of the RAS components, prorenin/renin, angiotensin-converting enzyme (ACE), angiotensin II, angiotensin II type 1 receptor (AT1R), and angiotensin II type 2 receptor (AT2R) in the normal mouse lacrimal gland. To examine the change in tear secretion, mice received ARB (AT1R blocker) or AT2R antagonist. Tear secretion was assessed by cotton thread test before and after drug administration. RESULTS. The mRNAs coding for angiotensinogen, prorenin, ACE, and both AT1R and AT2R were found in normal lacrimal gland tissue and cultured lacrimal gland fibroblasts. Prorenin/renin and ACE were identified in myoepithelial cells around ducts and acini and in blood vessels. Angiotensin II, AT1R, and AT2R were observed in the ducts and interstitial fibroblasts. AT1R and AT2R were also localized in blood vessels. All the cultured lacrimal gland fibroblasts expressed angiotensin II, AT1R, and AT2R. Tear secretion increased in mice that received ARB. CONCLUSIONS. The results are consistent with the hypothesis that a tissue-specific RAS is present in the lacrimal gland, and suggest that fibroblasts are one of the cell types playing a role in the tissue RAS. Tissue RAS might be involved in tissue function of regulating tear secretion in the lacrimal gland. © 2012 The Association for Research in Vision and Ophthalmology, Inc.

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Mitochondrial superoxide anion overproduction in Tet-mev-1 transgenic mice accelerates age-dependent corneal cell dysfunctions

Onouchi Hiromi, Ishii Takamasa, Miyazawa Masaki, Uchino Yuichi, Yasuda Kayo, Hartman Phil S., Kawai Kenji, Tsubota Kazuo, Ishii Naoaki

Investigative Ophthalmology and Visual Science 53 ( 9 ) 5780 - 5787 2012.08

ISSN 0146-0404

Purpose. The Tet-mev-1 mouse expressing a mitochondrial complex-II mutated SDHC^V69E gene controlled by a tetracycline (Tet)-On/Off system can overproduce O₂·^- and is a versatile whole-animal model for studying mitochondrial oxidative stress. Here we report a series of age-dependent variations in corneal epithelium, endothelium, and parenchymal cells of the Tet-mev-1 mice relative to wild-type C57BL/6j mice. Methods. Measurements of (1) mitochondrial electron transport enzyme activities; (2) O₂·^- production; (3) carbonylated protein, and 8-hydroxydeoxyguanosine (8-OHdG) levels as markers of oxidative stress; (4) pathologic analyses under optical and electron microscopy; (5) hematoxylin-eosin or toluidine-blue staining; and (6) immunohistochemistry with an anti-β-catenin antibody were performed in the eye, especially the cornea. Results. Complex II-III activity was decreased by electron leakage between complex II and CoQ. This resulted in increased age-dependent intracellular oxidative stress in the eye of Tet-mev-1 mice. Corneal epithelialization was delayed in Tet-mev-1 mice after 20% ethanol treatment, as the number of cells and mitotic cells decreased in the corneal epithelium of Tet-mev-1 mice compared with that of wild type. The age-dependent decrease in cell number accelerated in the corneal endothelium cells. Moreover, it was suggested that the corneal thickness was decreased by thinning of parenchymal cells with age in Tet-mev-1 mice. Conclusions. These results suggest that mitochondrial oxidative stress with electron transport chain dysfunction can influence pathogenesis and progression of age-related corneal diseases, as well as generalized corneal aging acceleration. © 2012 The Association for Research in Vision and Ophthalmology, Inc.

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Congenital corneal staphyloma as a complication of Kabuki syndrome

Tanaka Ryuma, Takenouchi Toshiki, Uchida Keiko, Sato Takeshi, Fukushima Hiroyuki, Yoshihashi Hiroshi, Takahashi Takao, Tsubota Kazuo, Kosaki Kenjiro

American Journal of Medical Genetics, Part A 158 A ( 8 ) 2000 - 2002 2012.08

ISSN 1552-4825

Kabuki syndrome has long been clinically defined based mainly on its characteristic eye features. The recent discovery of MLL2 as a causative gene of Kabuki syndrome has enabled the extreme end of the phenotype to be explored. We herein report on two patients with striking visible congenital staphyloma at birth. A diagnosis of Kabuki syndrome was subsequently made in both patients based on a constellation of characteristic eye features, cardiac abnormalities and severe developmental delay, and finally by the confirmation of MLL2 mutations. In conclusion, congenital corneal staphyloma is a complication of Kabuki syndrome with MLL2 mutations. © 2012 Wiley Periodicals, Inc..

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Intraocular pressure elevation after Descemet's stripping endothelial keratoplasty

Ozeki Naoki, Yuki Kenya, Shiba Daisuke, Shimmura Shigeto, Murat Dogru, Tsubota Kazuo

Japanese Journal of Ophthalmology 56 ( 4 ) 307 - 311 2012.07

ISSN 0021-5155

Purpose: To investigate the incidence of intraocular pressure (IOP) elevation after Descemet's stripping endothelial keratoplasty (DSEK) and associated clinical findings. Methods: Forty-eight eyes of 46 patients who underwent DSEK were studied in this single-center retrospective study. IOP elevation was defined as the presence of an increase in IOP above 21 mmHg lasting at least two - months, the need to introduce topical/oral antiglaucoma medications for at least two months, and the need for surgical therapy for uncontrolled IOP. Patients with pre-existing glaucoma or patients who were using antiglaucoma drugs before DSEK were excluded according to the protocol of the study. The incidence of post-DSEK IOP elevation was investigated. Results: The incidence of IOP elevation after DSEK was 16.7%. Glaucoma therapy following DSEK was needed in eight eyes of eight patients. IOP elevations were diagnosed from two to ten months after DSEK. There was no statistical difference in the survival probabilities of patients with different preoperative diagnoses. Pre-op IOP was observed to be a risk factor of IOP elevation in this series. Conclusions: IOP elevation appears to be a common postoperative complication of DSEK. Patients without preexisting glaucoma but with higher pre-op IOP need to be followed up carefully after DSEK. © Japanese Ophthalmological Society 2012.

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Topic of the month

Ayaki Masahiko, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 66 ( 7 ) 953 - 956 2012.07

ISSN 0370-5579

Low diastolic blood pressure is associated with the progression of normal-tension glaucoma

Okumura Yoshihiko, Yuki Kenya, Tsubota Kazuo

Ophthalmologica 228 ( 1 ) 36 - 41 2012.06

ISSN 0030-3755

Aims: The objective of this study was to investigate the association between blood pressure (BP) and visual field defect progression in normal-tension glaucoma (NTG). Methods: The subjects were 35 consecutive patients with NTG who were admitted to Keio University Hospital for 24-hour intraocular pressure evaluation; all subjects underwent 6 or more visual field tests in either eye and were followed up for more than 5 years. Systolic and diastolic BPs were measured at 9.00 h. Subjects with negative regression lines and p values of less than 0.05 were classified as progressive; all others were defined as stable. Systolic BP, diastolic BP, and ocular perfusion pressure were compared between the 2 groups. Results: Twelve subjects showed progression in glaucomatous visual field defects (age 55.1 ± 15.1 years). Twenty-three subjects showed no progression (age 57.2 ± 10.9 years). Diastolic BP (stable group: 77.0 ± 12.1 mm Hg; progressive group: 67.7 ± 9.7 mm Hg; p = 0.03) was significantly lower in the progressive group than in the stable group. Multivariable analysis revealed that low diastolic BP is a significant risk factor for progression (OR 0.90; 95% CI 0.82-0.99). Conclusions: Low diastolic BP was associated with glaucomatous visual field progression in subjects with NTG. Copyright © 2012 S. Karger AG, Basel.

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Characterization of cultivated murine lacrimal gland epithelial cells

Kobayashi Shinya, Kawakita Tetsuya, Kawashima Motoko, Okada Naoko, Mishima Kenji, Saito Ichiro, Ito Masataka, Shimmura Shigeto, Tsubota Kazuo

Molecular Vision 18 1271 - 1277 2012.05

ISSN 1090-0535

Purpose: To date, mouse lacrimal gland epithelial cells have been cultured successfully but only in cases involving newborn mouse lacrimal glands. In this work, we attempted to cultivate and characterize adult mouse lacrimal gland epithelial cells. Methods: Lacrimal glands were removed from newborn mice (C57B/6) and isolated lacrimal gland epithelial cells were seeded onto tissue culture treated or low adherent culture dishes in Cnt-07 culture medium with or without cholera toxin. Cultivated cells were characterized by immunostaining with pan-cytokeratin, α-smooth muscle actin, and lactoferrin antibodies. Lacrimal gland cells from 7-week-old green fluorescent protein (GFP) and non-GFP (C57B/6) mice were mixed and seeded onto uncoated dishes to assess sphere-forming efficiency. Cells were also seeded onto 3T3 cell feeder layers to assess colony forming efficiency. Results: Lacrimal gland epithelial cells were selectively cultured with cholera toxin, and cell type was verified by pancytokeratin and α-smooth muscle actin immunostaining. Sphere formation from single cells of adult mice was observed using specific medium and low adherent culture dishes. These cells could also undergo colony formation on 3T3 feeder cells. Conclusions: Adult mouse lacrimal gland epithelial cells were successfully cultivated in cholera toxin-containing medium, and were observed to form spheres from single cells. © 2012 Molecular Vision.

Punctal plugs for treatment of post-LASIK dry eye

Yung Yang Hao, Toda Ikuko, Sakai Chikako, Yoshida Atsushi, Tsubota Kazuo

Japanese Journal of Ophthalmology 56 ( 3 ) 208 - 213 2012.05

ISSN 0021-5155

Purpose: To evaluate the efficacy of punctal plugs for patients with post-LASIK dry eye. Materials and methods: A prospective study was conducted on 25 eyes of 18 patients who underwent LASIK. All eyes fulfilled the Japanese dry-eye criteria and had not responded to conventional treatment with artificial tears by 1 month postsurgery. They were randomly divided into a plug and a non-plug group. Punctal plugs were inserted into the superior and inferior puncta in the plug group after the 1-month examination. Uncorrected and best corrected visual acuity (UCVA, BCVA), refraction, functional visual acuity (FVA) after 10 s eye opening (FVA 10), surface regularity index (SRI) after 10 s eye opening (SRI 10), corneal sensitivity, Schirmer test, fluorescein staining (FS), tear break-up time (TBUT), and symptoms were compared between the groups 1 and 3 months postsurgery. Changes in these data from 1 to 3 months were also compared between the groups. Results: The values in SRI 10, FS, and TBUT were significantly better in the plug group at 3 months. Changes in the values of UCVA, FVA 10, FS, TBUT, and symptoms between 1 and 3 months improved significantly in the plug group compared with the non-plug group. Conclusion: Our results suggest that punctal occlusion for both puncta is effective for patients with post-LASIK dry eye that cannot be controlled by artificial tears alone. Punctal plugs can improve not only the symptoms and tear function, but also the quality of vision, for example FVA. © Japanese Ophthalmological Society 2012.

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Age-related dysfunction of the lacrimal gland and oxidative stress

Kojima Takashi, Wakamatsu Tais H., Dogru Murat, Ogawa Yoko, Igarashi Ayako, Ibrahim Osama M A, Inaba Takaaki, Shimizu Takahiko, Noda Setsuko, Obata Hiroto, Nakamura Shigeru, Wakamatsu Alda, Shirasawa Takuji, Shimazaki Jun, Negishi Kazuno, Tsubota Kazuo

American Journal of Pathology 180 ( 5 ) 1879 - 1896 2012.05

ISSN 0002-9440

An imbalance between free radical generation and radical scavenging antioxidant systems results in oxidative stress, which has been associated with cell injury observed in many age-related diseases. The superoxide dismutase (SOD) family is a major antioxidant system, and deficiency of Cu,Zn-superoxide dismutase-1 (Sod1) in mice leads to many different phenotypes that resemble accelerated aging. In this study we examined the morphologic features and the secretory functions of the lacrimal glands in Sod1 ^-/- mice. Lacrimal glands showed atrophy of acinar units; fibrosis; infiltration with CD4 ⁺ T cells, monocytes, and neutrophils; increased staining with both 4-hydroxy-2-nonenal and 8-hydroxy-2′-deoxyguanosine; increases in apoptotic cells; and the presence of the epithelial-mesenchymal transition in senescent Sod1 ^-/- mice. Electron microscopy findings revealed evidence of epithelial-mesenchymal transition, presence of swollen and degenerated mitochondria, and the presence of apoptotic cell death in the lacrimal glands of senescent Sod1 ^-/- mice. These alterations were also associated with the accumulation of secretory vesicles in acinar epithelial cells, decreased production of both stimulated and nonstimulated tears, and a decline in total protein secretion from the lacrimal glands. Our results suggest that Sod1 ^-/- mice may be a good model system in which to study the mechanism of reactive oxygen species-mediated lacrimal gland alterations. © 2012 American Society for Investigative Pathology.

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Improvement of accommodation with anti-oxidant supplementation in visual display terminal users

Uchino Y., Uchino M., Dogru M., Fukagawa K., Tsubota K.

Journal of Nutrition, Health and Aging 16 ( 5 ) 478 - 481 2012.05

ISSN 1279-7707

Objective: To determine the antioxidant supplementation effect on accommodation among VDT users. Design: a double blind randomized placebo controlled study. registered under clinical Trials.gov identifier no. ncT00877201. Participants and Controls: Fourty right eyes of 40 healthy VDT users (30 females, 10 males, mean age: 43.8±2.8 years, range: 40-49 years). 20 subjects (15 females, 5 males; mean age: 44.0±2.7 years, range: 40-49 years). Methods: subjects were required to take an antioxidant supplement, 20 age and sex matched subjects (15 females, 5 males; mean age: 43.6±3.1 years, range: 40-49 years) were required to take placebo medication for 4 weeks. Results: The mean of the change in accommodation power was significantly higher in the group receiving antioxidant supplements (0.20±0.50 Diopter (D)) compared to the placebo group (-0.12±0.48 (D)) (p<0.05). Conclusions: antioxidant supplementation was observed to improve accommodation in Japanese Visual Display Terminal (VDT) Users.

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OCT assessment of tear meniscus after punctal occlusion in dry eye disease

Ibrahim Osama M A, Dogru Murat, Kojima Takashi, Matsumoto Yukihiro, Wakamatsu Tais Hitomi, Tsubota Kazuo, Fujishima Hiroshi

Optometry and Vision Science 89 ( 5 ) 2012.05

ISSN 1040-5488

PURPOSE.: To evaluate the tear meniscus changes after punctal occlusion in dry eye patients using Visante optical coherence tomography (OCT) in a prospective controlled study. METHODS.: Thirty eyes of 15 symptomatic dry eye patients not responding to non-preserved artificial tears received additional upper and lower punctal occlusion with silicone plugs, and 30 eyes of 15 age-and sex-matched dry eye control patients received only non-preserved artificial tears treatment for 1 month. All subjects underwent tear meniscus height (TMH) measurements with Visante OCT. All study participants also underwent slitmicroscopy graticule scale TMH measurement, strip meniscometry testing, tear film break-up time measurement, ocular surface vital staining with fluorescein and Rose Bengal dyes, and the Schirmer-1 test. Both groups, dry eye and control group patients, were examined before and after 1-month treatment. Wilcoxon-matched pair test was performed. The study was conducted in compliance with the Tenets of the Declaration of Helsinki. RESULTS.: TMH measurements by OCT and slitlamp graticule scale significantly improved after punctal occlusion (p < 0.001) and remain unchanged in the dry eye control patients. Similarly, strip meniscometry scores, mean tear stability values, Rose Bengal, and fluorescein staining scores showed significant improvement after punctal occlusion (p < 0.05). Schirmer-1 test values tended to be higher after 1 month of treatment in both groups without any statistical significance (p > 0.05). CONCLUSIONS.: OCT TMH measurement appears to be effective in monitoring tear meniscus changes after punctal occlusion. OCT can be a valuable non-invasive and quick clinical tool for evaluation of treatment responses in dry eye patients. Copyright © 2012 American Academy of Optometry.

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Biological role of lutein in the light-induced retinal degeneration

Sasaki Mariko, Yuki Kenya, Kurihara Toshihide, Miyake Seiji, Noda Kosuke, Kobayashi Saori, Ishida Susumu, Tsubota Kazuo, Ozawa Yoko

Journal of Nutritional Biochemistry 23 ( 5 ) 423 - 429 2012.05

ISSN 0955-2863

Lutein, a xanthophyll of a carotenoid, is anticipated as a therapeutic product to prevent human eye diseases. However, its biological mechanism is still unclear. Here, we show the molecular mechanism of lutein's effect to reduce photodamage of the retina. We analyzed the light-exposed retinas of Balb/c mice given lutein-supplemented or normal diet. Visual function was measured by electroretinogram, and histological changes were observed. Immunohistochemical and immunoblot analyses were performed to analyze molecular mechanism. The reactive oxygen species induced in the retina was evaluated by fluorescent probes. In the mice after light exposure, reduction of a-wave and b-wave amplitudes in electroretinogram, indicating visual impairment, and thinning of the photoreceptor cell layer owing to apoptosis were both attenuated by lutein diet. Interestingly, γ-H2AX, a marker for double-strand breaks (DSBs) in DNA, was up-regulated in the photoreceptor cells after light exposure, but this increase was attenuated by lutein diet, suggesting that DSBs caused by photodamage contributed to the photoreceptor cell death and that this change was suppressed by lutein. Moreover, the expression of eyes absent (EYA), which promotes DNA repair and cell survival, was significantly up-regulated with lutein diet in the light-exposed retina. Therefore, lutein induced EYA for DNA repair, which could suppress DNA damage and photoreceptor cell apoptosis. Lutein reduced light-induced oxidative stress in the retina, which might contribute to promote DNA repair. The lutein-supplemented diet attenuated light-induced visual impairment by protecting the photoreceptor cells' DNA. © 2012 Elsevier Inc.

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Corneal and retinal effects of ultraviolet-B exposure in a soft contact lens mouse model

Ibrahim Osama M A, Kojima Takashi, Wakamatsu Tais Hitomi, Dogru Murat, Matsumoto Yukihiro, Ogawa Yoko, Ogawa Junko, Negishi Kazuno, Shimazaki Jun, Sakamoto Yasuo, Sasaki Hiroshi, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 53 ( 4 ) 2403 - 2413 2012.04

ISSN 0146-0404

PURPOSE. To investigate the lipid and DNA oxidative stress as well as corneal and retinal effects after ultraviolet B (UV-B) exposure in mice, with or without silicon hydrogel soft contact lenses (SCL). METHODS. Twenty-eight C57BL6-strain male mice were divided into four groups: group I, control group with no SCL (SCL [-]) and no UV-B exposure (UV-B [-]); group II, senofilcon A SCL (senofilcon [+]) with UV-B exposure (UV-B [+]); group III, lotrafilcon A SCL (lotrafilcon [+]) with UV-B exposure (UV-B [+]); and group IV, no SCL (SCL [-]), but with UV-B exposure (UV-B [+]). All mice except group I received UV-B exposure for 5 days for a total dose of 2.73 J/cm ². All mice underwent tear hexanoyl-lysine (HEL) and tear cytokine ELISA measurements, and fluorescein and rose bengal corneal staining before and after UV-B exposure. Corneal specimens underwent immunohistochemistry staining with CD45, HEL, 4-hydroxynonenal (4-HNE), and 8-hydroxy-20-deoxyguanosine (8-OHdG) antibodies and evaluation with electron microscopy. RESULTS. All mice without SCL but exposed to UV-B developed corneal edema, ulcers, or epithelial damage compared with mice with senofilcon A SCL and exposure to UV-B. Tear HEL and cytokine levels significantly increased in mice without SCL after UV-B exposure. Immunohistochemistry showed a significantly higher number of cells positively stained for CD45, 8-OHdG, HEL, and 4-HNE in the corneas of mice without SCLs compared with those with senofilcon A after UV-B exposure. CONCLUSION. Silicon hydrogel SCL showed corneal and retinal protective effects, owing to UV blocking properties, against oxidative stress-related membrane lipid and cellular DNA damage. © 2012 The Association for Research in Vision and Ophthalmology, Inc.

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Dietary lactoferrin alleviates age-related lacrimal gland dysfunction in mice

Kawashima Motoko, Kawakita Tetsuya, Inaba Takaaki, Okada Naoko, Ito Masataka, Shimmura Shigeto, Watanabe Mitsuhiro, Shinmura Ken, Tsubota Kazuo

PLoS ONE 7 ( 3 ) 2012.03

Background: Decrease in lacrimal gland secretory function is related to age-induced dry eye disease. Lactoferrin, the main glycoprotein component of tears, has multiple functions, including anti-inflammatory effects and the promotion of cell growth. We investigated how oral administration of lactoferrin affects age-related lacrimal dysfunction. Methods and Findings: Twelve-month-old male C57BL/6Cr Slc mice were randomly divided into a control fed group and an oral lactoferrin treatment group. Tear function was measured at a 6-month time-point. After euthanasia, the lacrimal glands were subjected to histological examination with 8-hydroxy-2′-deoxyguanosine (8-OHdG) antibodies, and serum concentrations of 8-OHdG and hexanoyl-lysine adduct (HEL) were evaluated. Additionally, monocyte chemotactic protein-1(MCP-1) and tumor necrosis factor-α (TNF-α) gene expression levels were determined by real-time PCR. The volume of tear secretion was significantly larger in the treated group than in the control. Lactoferrin administration reduced inflammatory cell infiltration and the MCP-1 and TNF-α expression levels. Serum concentrations of 8-OHdG and HEL in the lactoferrin group were lower than those in the control group and were associated with attenuated 8-OHdG immunostaining of the lacrimal glands. Conclusion: Oral lactoferrin administration preserves lacrimal gland function in aged mice by attenuating oxidative damage and suppressing subsequent gland inflammation. © 2012 Kawashima et al.

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Blockade of vascular adhesion protein-1 attenuates choroidal neovascularization

Yoshikawa Nami, Noda Kousuke, Ozawa Yoko, Tsubota Kazuo, Mashima Yukihiko, Ishida Susumu

Molecular Vision 18 593 - 600 2012.03

Purpose: Vascular adhesion protein (VAP)-1 is an adhesion molecule elucidated as a mediator of the leukocyte recruitment cascade. The purpose of this study was to investigate the role of VAP-1 in ocular inflammatory neovascularization using a mouse laser-induced choroidal neovascularization (CNV) model. Methods: CNV was induced with 532 nm laser irradiation in C57BL/6 mice, and production of VAP-1 protein in the retinal pigment epithelium (RPE) choroid during CNV formation was examined. CNV animals were treated with the specific VAP-1 inhibitor U-V002 or vehicle solution, and the volume of CNV tissue was evaluated with volumetric measurements. Macrophage infiltration into the CNV lesions was evaluated using two different techniques, flatmount staining and real-time polymerase chain reaction (PCR) for F4/80. The protein levels of intercellular adhesion molecule (ICAM)-1, monocyte chemoattractant protein (MCP)-1, P-selectin, and vascular endothelial growth factor (VEGF) in the RPE-choroid were measured with enzyme-linked immunosorbent assay (ELISA). Results: VAP-1 inhibition significantly suppressed CNV formation in a dose-dependent manner and reduced macrophage infiltration into CNV lesions. Furthermore, VAP-1 blockade decreased the expression of ICAM-1 and MCP-1, both of which play a pivotal role in macrophage recruitment. Conclusions: Our data suggest VAP-1 has an important role during ocular inflammatory neovascularization through leukocyte recruitment. VAP-1 inhibition may be a novel and potent therapeutic strategy in treating CNV formation. © 2012 Molecular Vision.

The association between primary open-angle glaucoma and fall

Tanabe Sachiko, Yuki Kenya, Ozeki Naoki, Shiba Daisuke, Tsubota Kazuo

Clinical Ophthalmology 6 ( 1 ) 327 - 331 2012.03

ISSN 1177-5467

Purpose: Falls are among the most serious public health concerns for the elderly. Information conveyed via the visual sense is relevant to postural balance and movement, and proper visual function is essential to avoid falls. Here we investigated the prevalence of injurious falls among patients with primary open-angle glaucoma (POAG) who were more than 45 years old, compared with comparably aged healthy subjects. Methods: This is a cross-sectional study. Consecutive patients who visited the Tanabe Eye Clinic, Yamanashi, Japan between January 1 and March 30, 2009 were screened for eligibility by ophthalmic examination. A total of 117 control subjects (77 men, 40 women; aged 60.2 ± 7.5 years) who were free of ocular disease and 101 POAG patients (58 men, 43 women; aged 62.3 ± 8.7 years) were consecutively enrolled. Participants answered a questionnaire on injurious fall experience during the previous 10 years. The prevalence of injurious fall in subjects with POAG versus healthy controls was examined with Fisher's exact test. Adjusted odds ratios and 95% confidence intervals were estimated with logistic regression models for the subjects with POAG (factors: age, gender, mean deviation in the better eye or worse eye). Results: The self-reported prevalence of injurious fall was 0.9% (1/117) in the control group and 6.9% (7/101) in the POAG group. The association between injurious fall and POAG was statistically significant (P = 0.026, Fisher's exact test). Within the POAG patients, the group reporting falls was significantly older and had a lower BMI, worse BCVA, and worse mean deviation in both the better and worse eye than the group reporting no falls. Worse mean deviation in the eye with the better visual field (odds ratios 0.75; 95% confidence intervals: 0.57 to 0.99; P = 0.036) was a significant risk factor for injurious falls in subjects with POAG. Conclusions: POAG was significantly associated with injurious falls. © 2012 Tanabe et al, publisher and licensee Dove Medical Press Ltd.

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The formation of an angiogenic astrocyte template is regulated by the neuroretina in a HIF-1-dependent manner

Nakamura-Ishizu Ayako, Kurihara Toshihide, Okuno Yuji, Ozawa Yoko, Kishi Kazuo, Goda Nobuhito, Tsubota Kazuo, Okano Hideyuki, Suda Toshio, Kubota Yoshiaki

Developmental Biology 363 ( 1 ) 106 - 114 2012.03

ISSN 0012-1606

The vascular and nervous systems display a high degree of cross-talk and depend on each other functionally. In the vascularization of the central nervous system, astrocytes have been thought to sense tissue oxygen levels in hypoxia-inducible factors (HIFs)-dependent manner and control the vascular growth into the hypoxic area by secreting VEGF. However, recent genetic evidences demonstrate that not only astrocyte HIFs but also astrocyte VEGF expression is dispensable for developmental angiogenesis of the retina. This study demonstrates that hypoxia-inducible factor 1 alpha subunit (HIF-1α), a key transcription factor involved in cellular responses to hypoxia, is most abundantly expressed in the neuroretina, especially retinal progenitor cells (RPCs). A neuroretina-specific knockout of HIF-1α (αCre ⁺Hif1α ^flox/flox) showed impaired vascular development characterized by decreased tip cell filopodia and reduced vessel branching. The astrocyte network was hypoplastic in αCre ⁺Hif1α ^flox/flox mice. Mechanistically, platelet-derived growth factor A (PDGF-A), a mitogen for astrocytes, was downregulated in the neuroretina of αCre ⁺Hif1α ^flox/flox mice. Supplementing PDGF-A restored reduced astrocytic and vascular density in αCre ⁺Hif1α ^flox/flox mice. Our data demonstrates that the neuroretina but not astrocytes acts as a primary oxygen sensor which ultimately controls the retinal vascular development by regulating an angiogenic astrocyte template. © 2011 Elsevier Inc.

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Mucosal microvilli in dry eye patients with chronic GVHD

Tatematsu Y., Ogawa Y., Shimmura S., Dogru M., Yaguchi S., Nagai T., Yamazaki K., Kameyama K., Okamoto S., Kawakami Y., Tsubota K.

Bone Marrow Transplantation 47 ( 3 ) 416 - 425 2012.03

ISSN 0268-3369

The ocular surface is a frequent target tissue of mucosal chronic GVHD (cGVHD). We investigated the histopathological features of the conjunctival microvilli in patients with cGVHD. Conjunctival tissue specimens from patients with cGVHD or Sjögren's syndrome (SS) or from healthy individuals were examined by light microscopy and EM, impression cytology, and immunohistochemistry. The cGVHD conjunctivae showed significantly more metaplasia and fewer goblet cells than the SS and normal conjunctivae. Abundant CD8 ⁺ T cells infiltrated the basal epithelia in the cGVHD conjunctiva. The microvilli per standard epithelial unit and the secretory vesicles were counted by analyzing electron micrographs. The mean number of mucosal microvilli was significantly lower in the cGVHD than that in the SS or normal specimens, and the microvilli were significantly shorter, with a smaller height-width ratio. The mean number of secretory vesicles was also significantly lower, and the membrane-spanning mucin thinner, in the cGVHD compared with the SS and normal specimens. Thus, the conjunctival mucosal microvilli of cGVHD patients were significantly different in number and morphology from those of SS and normal subjects. These may be important factors affecting the stability of the tear-film layer and its contribution to cGVHD-related dry eye. © 2012 Macmillan Publishers Limited All rights reserved.

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The Vision van, a mobile eye clinic, Aids relief Efforts in Tsunami-Stricken areas

Oshima Catherine Ross, Yuki Kenya, Uchida Atsuro, Dogru Murat, Koto Takashi, Ozawa Yoko, Tsubota Kazuo

Keio Journal of Medicine 61 ( 1 ) 10 - 14 2012.03

ISSN 0022-9717

In the wake of the devastating earthquake and tsunami of March 11, our group at Keio University worked diligently to bring the Vision Van, a mobile eye clinic, from Miami, Florida, to disaster areas where medicines, eyeglasses, and indeed all other ophthalmological supplies and services, were nonexistent, thereby assisting many tsunami survivors suffering from eye problems. This act was encouraging not only to the patients and those working in the disaster-hit areas but also to ophthalmologists who wanted to use their medical specialty to assist those in need. © 2012 by The Keio Journal of Medicine.

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Use of micronutrient supplement for preventing advanced age-related macular degeneration in Japan

Sasaki Mariko, Shinoda Hajime, Koto Takashi, Uchida Atsuro, Tsubota Kazuo, Ozawa Yoko

Archives of Ophthalmology 130 ( 2 ) 254 - 255 2012.02

ISSN 0003-9950

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A novel clinical entity, IgG4-relateddisease (IgG4RD)

Umehara Hisanori, Okazaki Kazuichi, Masaki Yasufumi, Kawano Mitsuhiro, Yamamoto Motohisa, Saeki Takako, Matsui Shoko, Sumida Takayuki, Mimori Tsuneyo, Tanaka Yoshiya, Tsubota Kazuo, Yoshino Tadashi, Kawa Shigeyuki, Suzuki Ritsuro, Takegami Tsutomu, Tomosugi Naohisa, Kurose Nozomu, Ishigaki Yasuhito, Azumi Atsushi, Kojima Masaru, Nakamura Shigeo, Inoue Dai, Fujikawa Keita, Hamano Hideaki, Kubo Keiji, Yamamoto Hiroshi, Hirokawa Mitsuyosi, Itoh Kunihiko, Kamisawa Terumi, Kawabata Daisuke, Matsumoto Morio, Minamoto Seijiro, Murayama Kayoko, Nishiyama Susumu, Ogawa Yoko, Origuchi Tomoki, Oyama Norihide, Sato Yasuharu, Seto Masao, Sugai Susumu, Tsukamoto Norifumi, Takahira Masayuki, Takahashi Hiroki, Tsuboi Hiroto, Waseda Yuko, Kitagawa Kazuko, Nojima Takayuki, Yokoyama Hitoshi, Tonami Hisao, Fukushima Toshihiro, Tanaka Masao, Fujita Yoshimasa, Sawaki Toshioki, Kawanami Takafumi, Miki Miyuki, Iwao Haruka, Nakajima Akio, Nakamura Takuji

Modern Rheumatology 22 ( 1 ) 1 - 14 2012.02

ISSN 1439-7595

IgG4-related disease (IgG4RD) is a novel clinical disease entity characterized by elevated serum IgG4 concentration and tumefaction or tissue infiltration by IgG4-positive plasma cells. IgG4RD may be present in a certain proportion of patients with a wide variety of diseases, including Mikulicz's disease, autoimmune pancreatitis, hypophysitis, Riedel thyroiditis, interstitial pneumonitis, interstitial nephritis, prostatitis, lymphadenopathy, retroperitoneal fibrosis, inflammatory aortic aneurysm, and inflammatory pseudotumor. Although IgG4RD forms a distinct, clinically independent disease category and is attracting strong attention as a new clinical entity, many questions and problems still remain to be elucidated, including its pathogenesis, the establishment of diagnostic criteria, and the role of IgG4. Here we describe the concept of IgG4RD and up-to-date information on this emerging disease entity. © Japan College of Rheumatology 2011.

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Simple and accurate alignment of toric intraocular lenses and evaluation of their rotation errors using anterior segment optical coherence tomography

Watanabe Kazuhiro, Negishi Kazuno, Torii Hidemasa, Saiki Megumi, Dogru Murat, Tsubota Kazuo

Japanese Journal of Ophthalmology 56 ( 1 ) 31 - 37 2012.01

ISSN 0021-5155

Purpose: To describe a new method of alignment of toric intraocular lenses (IOLs) and evaluation of their rotation errors using anterior segment optical coherence tomography (AS-OCT). Methods: Twenty-nine eyes of 22 patients who had cataract extraction and implantation of an acrylic toric IOL were included. The new AS-OCT method was used for the alignment of toric IOLs and evaluation of their rotation errors. These rotation errors were evaluated and compared with those measured using the internal map of a wavefront aberrometer. Results: The mean rotation error ± standard deviation (SD) of the toric IOLs evaluated by AS-OCT was 3.2 ± 2.2° and 3.2 ± 2.4° at 1 week and 1 month after surgery, respectively. The mean difference in the reference axis between the visits was 1.8 ± 2.1°. The mean difference between the rotation errors of the alignment axes measured by AS-OCT and the internal map was 2.5 ± 1.9° (P = 0.037). Conclusion: The current method is clinically useful not only for the accurate alignment of toric IOLs but also for evaluating their rotation errors. © Japanese Ophthalmological Society 2011.

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Comparison of stem cell sources in the severity of dry eye after allogeneic haematopoietic stem cell transplantation

Uchino Miki, Ogawa Yoko, Uchino Yuichi, Mori Takehiko, Okamoto Shinichiro, Tsubota Kazuo

British Journal of Ophthalmology 96 ( 1 ) 34 - 37 2012.01

ISSN 0007-1161

Aims: To compare the incidence and severity of dry eye (DE) after allogeneic haematopoietic stem cell transplantation (HSCT) according to the stem cell source. The authors specifically focused on patients who received bone marrow transplantation (BMT), peripheral blood stem cell transplantation (PBSCT) and cord blood transplantation (CBT). Methods: Ninety-nine HSCT recipients who were prospectively followed-up for at least 100 days at Keio University Hospital were recruited. Ophthalmological examinations included evaluation of ocular surface findings and tear dynamics. The data on systemic graft-versus-host disease were collected by chart review. Results: Of the 99 patients (BMT, 67; PBSCT, 18; CBT, 14), 42 developed DE or showed worsened pre-existing DE after HSCT; 31 (46.3%) BMT group; 8 (44.0%) PBSCT group; and 3 (21.4%) CBT group (p=0.78). The median onset time of DE tended to be later in the PBSCT group (474 days, range 95-1559) than in the BMT (287 days, range 67-1216) or CBT (168 days, range 33-481) group, but the difference was not significant (p=0.23). However, the proportion of patients with severe DE was significantly higher in the PBSCT group (N=7, 87.5%) than in the BMT (N=12, 38.7%) or CBT (N=1, 33.3%) group (p=0.04) and CBT showed the lowest among all three stem cell sources. Conclusion: The data in this study suggested that the severity and onset time of DE were affected by the stem cell source. Close attention must be paid to the development of late-onset severe DE in PBSCT recipients.

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Changes in dry eye diagnostic status following implementation of revised Japanese dry eye diagnostic criteria

Uchino Yuichi, Uchino Miki, Dogru Murat, Ward Samantha, Yokoi Norihiko, Tsubota Kazuo

Japanese Journal of Ophthalmology 56 ( 1 ) 8 - 13 2012.01

ISSN 0021-5155

Purpose: Our aim was to evaluate changes in dry eye diagnostic status following implementation of the new dry eye diagnostic criteria in Japan. Design: This was a multicenter cross-sectional study. Methods: We recruited 295 individuals (81 men, 214 women, average age 43.6 ± 14.3 years) seen for general ophthalmic checkup and dry eye examinations. Using results of the Schirmer I test, tear breakup time, and fluorescein and Rose Bengal staining, patients were diagnosed as having definite dry eye (DDE), probable dry eye (PDE), or as being normal according to both the old and new Japanese dry eye diagnostic criteria. Results: Mean ages of normal participants and patients with PDE and DDE were 37.0 ± 10.4, 41.7 ± 14.4, and 47.7 ± 15.3 years, respectively (p < 0.001). All 37 individuals diagnosed as normal following the old criteria were also diagnosed as normal with the new diagnostic criteria. Among the 60 patients diagnosed as PDE with the old criteria, 19 (31.7%) were diagnosed as normal and 41(68.3%) as PDE with the new diagnostic criteria. Of the 198 patients diagnosed with DDE following the old criteria, 59 (29.7%) were diagnosed as PDE and 139 (70.2%) as DDE with the new diagnostic criteria. There was no significant difference in dry eye severity index scores between the old and new diagnostic criteria. Conclusion: A shift in the final dry eye diagnostic status from DDE to PDE and from PDE to normal was observed with the implementation of the new dry eye diagnostic criteria, suggesting that patients at the severe end of the dry eye disease spectrum are now diagnosed as DDE disease under the new criteria. © Japanese Ophthalmological Society 2011.

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Vision preservation during retinal inflammation by anthocyanin-rich bilberry extract

Miyake Seiji, Takahashi Noriko, Sasaki Mariko, Kobayashi Saori, Tsubota Kazuo, Ozawa Yoko

Laboratory Investigation 92 ( 1 ) 102 - 109 2012.01

ISSN 0023-6837

Anthocyanin-rich bilberry extract, a plant-derived antioxidant, has been utilized as a popular supplement for ocular health worldwide. However, it is unclear whether this extract has any biological effect on visual function, and the mechanism for such an effect is completely unknown. In this study, we generated a mouse model of endotoxin-induced uveitis (EIU) that shows retinal inflammation, as well as uveitis, by injecting lipopolysaccharide. We pretreated the mice with anthocyanin-rich bilberry extract and analyzed the effect on the retina. Anthocyanin-rich bilberry extract prevented the impairment of photoreceptor cell function, as measured by electroretinogram. At the cellular level, we found that the EIU-associated rhodopsin decreased and the shortening of outer segments in photoreceptor cells were suppressed in the bilberry-extract-treated animals. Moreover, the extract prevented both STAT3 activation, which induces inflammation-related rhodopsin decrease, and the increase in interleukin-6 expression, which activates STAT3. In addition to its anti-inflammatory effect, the anthocyanin-rich bilberry extract ameliorated the intracellular elevation of reactive oxygen species and activated NF-B, a redox-sensitive transcription factor, in the inflamed retina. Our findings indicate that anthocyanin-rich bilberry extract has a protective effect on visual function during retinal inflammation. © 2012 USCAP, Inc All rights reserved.

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Neuroprotective effects of lutein in the retina

Ozawa Yoko, Sasaki Mariko, Takahashi Noriko, Kamoshita Mamoru, Miyake Seiji, Tsubota Kazuo

Current Pharmaceutical Design 18 ( 1 ) 51 - 56 2012.01

ISSN 1381-6128

Although a large variety of pharmaceutical therapies for treating disease have been developed in recent years, there has been little progress in disease prevention. In particular, the protection of neural tissue is essential, because it is hardly regenerated. The use of nutraceuticals for maintaining the health has been supported by several clinical studies, including cross-sectional and interventional studies for age-related macular disease. However, mechanistic evidence for their effects at the molecular level has been very limited. In this review, we focus on lutein, which is a xanthophyll type of carotenoid. Lutein is not synthesized in mammals, and must be obtained from the diet. It is delivered to the retina, and in humans, it is concentrated in the macula. Here, we describe the neuroprotective effects of lutein and their underlying molecular mechanisms in animal models of vision-threatening diseases, such as innate retinal inflammation, diabetic retinopathy, and light-induced retinal degeneration. In lutein-treated mouse ocular disease models, oxidative stress in the retina is reduced, and its downstream pathological signals are inhibited. Furthermore, degradation of the functional proteins, rhodopsin (a visual substance) and synaptophysin (a synaptic vesicle protein also influenced in other neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease), the depletion of brain-derived neurotrophic factor (BDNF), and DNA damage are prevented by lutein, which preserves visual function. We discuss the possibility of using lutein, an antioxidant, as a neuroprotective treatment for humans. © 2012 Bentham Science Publishers.

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Plasma separation from human blood using spiral microchannels for dry eye treatment

Morikawa Jumpei, Yasui Takao, Kaji Noritada, Okamoto Yukihiro, Tokeshi Manabu, Tsubota Kazuo, Baba Yoshinobu

464 - 466 2012

Acute visual field defect following vitrectomy determined to originate from optic nerve by electrophysiological tests

Uchida Atsuro, Shinoda Kei, Matsumoto Celso Soiti, Kawai Miho, Kawai Sayaka, Ohde Hisao, Ozawa Yoko, Ishida Susumu, Inoue Makoto, Mizota Atsushi, Tsubota Kazuo

Case Reports in Ophthalmology 3 ( 3 ) 396 - 405 2012

Purpose: To present our findings on the cause of an acute visual field defect (VFD) that developed in a patient on the day after vitrectomy for proliferative diabetic retinopathy. Case: A 50-year-old man complained of a blind area in the superior visual field that developed one day after vitrectomy. The patient had undergone uncomplicated vitrectomy for a long-duration vitreous hemorrhage associated with proliferative diabetic retinopathy. Residual vitreous hemorrhage hampered a clear view of the fundus. Goldmann perimetry showed a horizontal VFD in the superior field. The area corresponding to the VFD was examined by multifocal electroretinograms (mfERGs) and multifocal visual evoked potentials (mfVEPs). The amplitudes of the mfVEPs were reduced with prolonged implicit times especially when the superior hemifield was stimulated, while the amplitudes and implicit times were within the normal range when other parts of the visual field were stimulated. In addition, the full-field photopic ERGs and photopic negative responses were attenuated in the right eye. These findings suggested that the VFD did not originate from alterations in the retinal inner and middle layer but in the ganglion cells. The visual acuity improved to 1.2 but his optic disc became pale and the VFD remained unchanged more than 12 years after the surgery. Conclusion: We suggest that vitrectomy can cause ischemic optic neuropathy by interfering with the circulation associated with diabetes mellitus. Evaluations by mfERGs, mfVEPs, and full-field photopic ERGs were helpful in making the diagnosis. © 2012 S. Karger AG, Basel.

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Transpupillary thermotherapy for atypical central serous chorioretinopathy

Kawamura Ryosuke, Ideta Hidenao, Hori Hideyuki, Yuki Kenya, Uno Tsuyoshi, Tanabe Tatsurou, Tsubota Kazuo, Kawasaki Tsutomu

Clinical Ophthalmology 6 ( 1 ) 175 - 179 2012

ISSN 1177-5467

Background: Central serous chorioretinopathy (CSC) has been traditionally treated with laser photocoagulation. We thought that transpupillary thermotherapy (TTT) utilizing a lower temperature than that of conventional laser photocoagulation might minimize permanent retinal and choroidal damage. Studies suggest that undesirable effects on vision due to TTT are minimal even if it is applied to foveal and/or parafoveal lesions when TTT requires a larger irradiation spot. The aim of this study was to evaluate the efficacy of TTT in the management of atypical CSC. Methods: We defined atypical CSC as bullous retinal detachment with diffuse or several leakages, severe leakage with fibrin formation under serous retinal detachment, or leakage within a pigment epithelium detachment. Eight consecutive patients with atypical CSC underwent visual acuity testing, ophthalmic examination, color photography, fluorescein angiography, and optical coherence tomography to evaluate the results of transpupillary thermotherapy. Retreatment of atypical CSC was based on ophthalmic examination, optical coherence tomography, and fluorescein angiography. TTT was performed on the leaking spots shown in fluorescein angiography, with a power of 50-250 mW, spot size of 500-1200 μm, and exposure time of 13-60 seconds to minimize retinal damage. Results: In five of eight affected eyes, serous detachments completely resolved within 1 month after the initial TTT. One eye had persistent subretinal fluid and required a second TTT treatment. Two eyes showed no resolution of CSC and were treated by conventional photocoagulation. Initial best-corrected visual acuity (BCVA) ranged from 20/600 to 20/20 (mean, 20/40; median, 20/30). Final BCVA ranged from 20/200 to 20/20 (mean, 20/25; median, 20/20). BCVA improved in all cases. Only two eyes with persistent subretinal fibrin and existing retinal pigment epithelial alternations in macular area showed limited improvement of BCVA despite the absence of subretinal exudation. The presence of retinal attachment was confirmed by optical coherence tomography in six eyes (75%). Conclusions: TTT seems to be effective for the treatment of atypical CSC in the short term. Additional studies are necessary to evaluate the long-term effectiveness and safety. © 2012 Kawamura et al, publisher and licensee Dove Medical Press Ltd.

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Renin-angiotensin system hyperactivation can induce inflammation and retinal neural dysfunction

Kurihara Toshihide, Ozawa Yoko, Ishida Susumu, Okano Hideyuki, Tsubota Kazuo

International Journal of Inflammation 2012 2012

ISSN 2090-8040

The renin-angiotensin system (RAS) is a hormone system that has been classically known as a blood pressure regulator but is becoming well recognized as a proinflammatory mediator. In many diverse tissues, RAS pathway elements are also produced intrinsically, making it possible for tissues to respond more dynamically to systemic or local cues. While RAS is important for controlling normal inflammatory responses, hyperactivation of the pathway can cause neural dysfunction by inducing accelerated degradation of some neuronal proteins such as synaptophysin and by activating pathological glial responses. Chronic inflammation and oxidative stress are risk factors for high incidence vision-threatening diseases such as diabetic retinopathy (DR), age-related macular degeneration (AMD), and glaucoma. In fact, increasing evidence suggests that RAS inhibition may actually prevent progression of various ocular diseases including uveitis, DR, AMD, and glaucoma. Therefore, RAS inhibition may be a promising therapeutic approach to fine-tune inflammatory responses and to prevent or treat certain ocular and neurodegenerative diseases. © 2012 Toshihide Kurihara et al.

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Neuroprotective response after photodynamic therapy

Suzuki Misa, Ozawa Yoko, Kubota Shunsuke, Hirasawa Manabu, Miyake Seiji, Noda Kousuke, Tsubota Kazuo, Kadonosono Kazuaki, Ishida Susumu

Journal of Neuroinflammation 8 2011.12

Background: Anti-vascular endothelial growth factor (VEGF) drugs and/or photodynamic therapy (PDT) constitute current treatments targeting pathological vascular tissues in tumors and age-related macular degeneration. Concern that PDT might induce VEGF and exacerbate the disease has led us to current practice of using anti-VEGF drugs with PDT simultaneously. However, the underlying molecular mechanisms of these therapies are not well understood.Methods: We assessed VEGF levels after PDT of normal mouse retinal tissue, using a laser duration that did not cause obvious tissue damage. To determine the role of PDT-induced VEGF and its downstream signaling, we intravitreally injected a VEGF inhibitor, VEGFR1 Fc, or a PI3K/Akt inhibitor, LY294002, immediately after PDT. Then, histological and biochemical changes of the retinal tissue were analyzed by immunohistochemistry and immunoblot analyses, respectively.Results: At both the mRNA and protein levels, VEGF was upregulated immediately and transiently after PDT. VEGF suppression after PDT resulted in apoptotic destruction of the photoreceptor cell layer in only the irradiated area during PDT. Under these conditions, activation of the anti-apoptotic molecule Akt was suppressed in the irradiated area, and levels of the pro-apoptotic protein BAX were increased. Intravitreal injection of a PI3K/Akt inhibitor immediately after PDT increased BAX levels and photoreceptor cell apoptosis.Conclusion: Cytotoxic stress caused by PDT, at levels that do not cause overt tissue damage, induces VEGF and activates Akt to rescue the neural tissue, suppressing BAX. Thus, the immediate and transient induction of VEGF after PDT is neuroprotective. © 2011 Suzuki et al; licensee BioMed Central Ltd.

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Corneal damage and lacrimal gland dysfunction in a smoking rat model

Higuchi Akihiro, Ito Kaori, Dogru Murat, Kitamura Masanori, Mitani Fumiko, Kawakita Tetsuya, Ogawa Yoko, Tsubota Kazuo

Free Radical Biology and Medicine 51 ( 12 ) 2210 - 2216 2011.12

ISSN 0891-5849

Smoking is a serious public health problem around the world and causes many diseases such as chronic obstructive pulmonary disease, lung cancer, and some eye diseases. Cytochrome P450s (CYPs) are xenobiotic-metabolizing enzymes and are distributed in the corneas, protecting the ocular surface against chemical compounds in the environment. Although CYPs are principally detoxification enzymes, CYP1A1 and CYP2A6 are known to participate in the induction of lung cancer by smoking. We studied the participation of CYPs in corneal dysfunction caused by exposure to mainstream cigarette smoke (MCS) in a smoking rat model. Six-week-old male Sprague-Dawley rats were exposed to MCS. Exposure to MCS caused corneal damage and lacrimal gland dysfunction. Immunohistochemical analysis revealed that CYP1A1 expression was upregulated in the corneal epithelium and ducts of the lacrimal glands, accompanied by an increase in production of reactive oxygen species (ROS). An increase in 8-hydroxy-2′- deoxyguanosine, which is a marker of oxidative DNA damage, was detected only in areas where CYP1A1 was expressed, whereas the level of hexanoyl-lysine adduct, which is an initial marker of oxidative damage of phospholipids, did not increase. Exposure to MCS damaged the corneas and lacrimal glands probably through DNA oxidation by ROS produced by CYP1A1. Although the influence of other components in MCS remains unclear, CYPs, especially CYP1A1, probably participate in corneal damage and lacrimal gland dysfunction induced by smoking. © 2011 Elsevier Inc. All rights reserved.

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Generation of stratified squamous epithelial progenitor cells from mouse induced pluripotent stem cells

Yoshida Satoru, Yasuda Miyuki, Miyashita Hideyuki, Ogawa Yoko, Yoshida Tetsu, Matsuzaki Yumi, Tsubota Kazuo, Okano Hideyuki, Shimmura Shigeto

PLoS ONE 6 ( 12 ) 2011.12

Background: Application of induced pluripotent stem (iPS) cells in regenerative medicine will bypass ethical issues associated with use of embryonic stem cells. In addition, patient-specific IPS cells can be useful to elucidate the pathophysiology of genetic disorders, drug screening, and tailor-made medicine. However, in order to apply iPS cells to mitotic tissue, induction of tissue stem cells that give rise to progeny of the target organ is required. Methodology/Principal Findings: We induced stratified epithelial cells from mouse iPS cells by co-culture with PA6 feeder cells (SDIA-method) with use of BMP4. Clusters of cells positive for the differentiation markers KRT1 or KRT12 were observed in KRT14-positive colonies. We successfully cloned KRT14 and p63 double-positive stratified epithelial progenitor cells from iPS-derived epithelial cells, which formed stratified epithelial sheets consisting of five- to six-polarized epithelial cells in vitro. When these clonal cells were cultured on denuded mouse corneas, a robust stratified epithelial layer was observed with physiological cell polarity including high levels of E-cadherin, p63 and K15 expression in the basal layer and ZO-1 in the superficial layer, recapitulating the apico-basal polarity of the epithelium in vivo. Conclusions/Significance: These results suggest that KRT14 and p63 double-positive epithelial progenitor cells can be cloned from iPS cells in order to produce polarized multilayer epithelial cell sheets. © 2011 Yoshida et al.

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Correlation between contrast sensitivity and higher-order aberration based on pupil diameter after cataract surgery

Yamaguchi Takefumi, Negishi Kazuno, Ohnuma Kazuhiko, Tsubota Kazuo

Clinical Ophthalmology 5 ( 1 ) 1701 - 1707 2011.12

ISSN 1177-5467

Background: The purpose of this study was to evaluate the correlation between contrast sensitivity and calculated higher-order aberrations based on individual natural pupil diameter after cataract surgery. Methods: This prospective study included 120 eyes from 92 patients who were randomized to receive one of four lenses, including three aspheric lenses (Acrysof SN60WF, Tecnis ZA9000, and Hoya Py60AD) and one spherical lens (Acrysof SN60AT). Contrast sensitivity, higher-order aberrations of the whole eye, and pupil diameter under photopic and mesopic conditions were measured 1 month postoperatively. Higher-order aberrations were decomposed into Zernike coefficients, calculated according to individual pupil diameter. The correlation between higher-order aberrations and contrast sensitivity was evaluated. Results: There were no significant differences in contrast sensitivity function between the four types of lenses under photopic conditions. However, the contrast sensitivity function and area under log contrast sensitivity function in the aspheric lenses were significantly better than in the spherical lens under mesopic conditions. Under mesopic conditions, spherical aberration in eyes with aspheric lenses was significantly lower than in eyes with spherical lenses (P,<0.05). Under photopic conditions, coma aberration had a significant negative correlation with contrast sensitivity at 12 cycles/degree. Under mesopic conditions, spherical aberration had a significant negative correlation with contrast sensitivity at 3, 6, and 12 cycles/degree with glare, and with contrast sensitivity at 6 and 18 cycles/degree without glare. Conclusion: In terms of influence on visual function, coma aberration may be more significant under photopic conditions and spherical aberration under mesopic conditions. © 2011 Yamaguchi et al, publisher and licensee Dove Medical Press Ltd.

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Prevalence and risk factors of dry eye disease in Japan

Uchino Miki, Nishiwaki Yuji, Michikawa Takehiro, Shirakawa Kazuhiro, Kuwahara Erika, Yamada Mutsuko, Dogru Murat, Schaumberg Debra A., Kawakita Tetsuya, Takebayashi Toru, Tsubota Kazuo

Ophthalmology 118 ( 12 ) 2361 - 2367 2011.12

ISSN 0161-6420

Objective: To estimate the prevalence and risk factors of dry eye disease (DED) in a rural setting in Japan. Design: Cross-sectional study. Participants: We included 3294 subjects, aged <40 years who were in the residential registry for Koumi town. Intervention: Subjects in a rural mountain area, Koumi town, completed questionnaires designed to detect dry eye diagnosis and risk factors. Main Outcome Measures: Clinically diagnosed DED was defined as the presence of a previous clinical diagnosis of DED by ophthalmologists or severe symptoms of DED (both dryness and irritation constantly or often). Current symptoms of DED and possible risk factors such as age, gender, educational history, smoking history, alcohol drinking history, height and weight, visual display terminal (VDT) use, and contact lens (CL) wear, and past/current history of certain common systemic diseases were the main outcome measures. We used logistic regression analysis to examine associations between DED and other demographic factors. Results: Of the 3294 eligible residents, 2791 residents (85%) completed the questionnaire. The percentage of women with a composite outcome of clinically diagnosed DED or severe symptoms (21.6%; 95% confidence interval [CI], 19.523.9) was higher than that of men (12.5%; 95% CI, 10.714.5; P<0.001). A low body mass index (BMI; odds ratio [OR], 2.07; 95% CI, 0.984.39), CL use (OR, 3.84; 95% CI, 1.4610.10), and hypertension (HT) (OR, 1.39; 95% CI, 0.942.06) were risk factors for DED in men. Use of a VDT (OR, 2.33; 95% CI, 1.124.85), CL use (OR, 3.61; 95% CI, 2.136.10), and myocardial infarction or angina were the risk factors (OR, 2.64; 95% CI, 1.514.62), whereas high BMI was a preventive factor (OR, 0.69; 95% CI, 0.481.01) for DED in women. Conclusions: Among a Japanese cohort, DED leading to a clinical diagnosis or severe symptoms is prevalent. Use of CLs was a common dry eye risk factor in both genders. The condition is more prevalent in men with low BMI, HT, and in women with myocardial infarction or angina and VDT use. Relevant measures directed against the modifiable risks may provide a positive impact on public health and quality of life of Japanese. © 2011 by the American Academy of Ophthalmology.

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Corneal fluorescein staining correlates with visual function in dry eye patients

Kaido Minako, Matsumoto Yukihiro, Shigeno Yuta, Ishida Reiko, Dogru Murat, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 13 ) 9516 - 9522 2011.12

ISSN 0146-0404

Purpose. To investigate the changes in functional visual acuity (VA) and higher order aberrations in dry eye patients. Methods. In this prospective comparative case series, 22 right eyes were classified into those with or without superficial punctate keratopathy (SPK) in the central cornea of 22 patients with Sjögren syndrome; 10 right eyes of 10 normal subjects served as the control. Serial measurements of VAs using a functional VA measurement system and higher order aberrations using a wavefront sensor were performed under blink-free conditions without topical anesthesia over a 10-second period. The parameters for each measurement were compared among the SPK-positive and -negative and normal groups. The correlation between those parameters was also analyzed. Results. Dry eye with SPK showed significant deterioration of visual function and optical quality compared with dry eye without SPK and in normal eyes, as detected by both the visual maintenance ratio (VMR; P < 0.05) and the variation of VA (P < 0.05) and by comalike and total higher order aberrations (P < 0.05). Moreover, the severity of epithelial damage at the central cornea correlated significantly with VMR (P < 0.01) and variation of VA (P < 0.01) as well as comalike (P < 0.05) and total higher order aberrations (P < 0.05). The dry eye group without SPK showed minor visual deterioration compared with normal eyes, as detected only by VMR (P < 0.05). Conclusions. Optical disturbances at the central optical zone of the cornea in dry eye disease may affect visual performance. Functional VA measurement may be an applicable method of evaluating visual performance in dry eyes that is as efficient as wavefront aberration measurements. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Inflammation after phakic implants

Torii Hidemasa, Negishi Kazuno, Dogru Murat, Saiki Megumi, Yamaguchi Takefumi, Watanabe Kazuhiro, Kato Naoko, Tsubota Kazuo

Ophthalmology 118 ( 12 ) 2011.12

ISSN 0161-6420

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The impact of contact lens wear and visual display terminal work on ocular surface and tear functions in office workers

Kojima Takashi, Ibrahim Osama M A, Wakamatsu Tais, Tsuyama Atsushi, Ogawa Junko, Matsumoto Yukihiro, Dogru Murat, Tsubota Kazuo

American Journal of Ophthalmology 152 ( 6 ) 2011.12

ISSN 0002-9394

Purpose: To evaluate the effect of contact lens (CL) wear and visual display terminal (VDT) work on the ocular surface and tear functions. Design: Prospective case-control study. Methods: Sixty-nine CL wearers (45 women and 24 men; mean age, 35.2 ± 7.3 years), and 102 age- and sex-matched non-CL wearers were enrolled in the study (66 women and 36 men; mean age, 36.7 ± 7.3 years). Ocular surface and tear function tests, including vital stainings (fluorescein and rose bengal), Schirmer test, tear meniscus height measurement, and tear film break-up time were performed. The subjective symptoms of dry eyes were evaluated using a dry eye symptom questionnaire. The participants were divided into 4 subgroups according to the total time of VDT work in 1 day (VDT work time in 1 day < 4 hours or < 4 hours) and presence of CL wear. Main outcome measures included ocular surface vital staining scores, Schirmer test results, tear film break-up time, tear meniscus height measurement, and symptom questionnaire score. Results: CL users and long-term VDT workers showed significantly worse tear meniscus height values than non-CL users and short-term VDT workers (P <.001). The mean visual symptom scores in CL wearers and long-term VDT workers were significantly higher than the other groups (P <.001). Conclusions: Office workers who wore CLs and spent more than 4 hours engaged in VDT work had a lower tear meniscus volume with significant dry eye and visual symptoms triggered by environmental factors. © 2011 Elsevier Inc.

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Miami to Japan eye-care rescue mission

Tsubota Kazuo, Yoshida Tsunehiko, Kurosaka Daijiro, Lee Richard K., Alfonso Eduardo C., Nakazawa Toru

American Journal of Ophthalmology 152 ( 5 ) 886 - 887 2011.11

ISSN 0002-9394

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Roles of AMP-activated protein kinase in diabetes-induced retinal inflammation

Kubota Shunsuke, Ozawa Yoko, Kurihara Toshihide, Sasaki Mariko, Yuki Kenya, Miyake Seiji, Noda Kousuke, Ishida Susumu, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 12 ) 9142 - 9148 2011.11

ISSN 0146-0404

Purpose. AMP-activated protein kinase (AMPK) is a sensor of cellular energy status. The purpose of the present study was to elucidate the roles of AMPK in the pathogenesis of diabetic retinopathy using the known AMPK activators resveratrol and AICAR (5-aminoimidazole-4-carboxamide ribonucleoside) in a mouse model. Methods. C57BL/6 mice with streptozotocin-induced diabetes were treated with resveratrol orally at 50 mg/kg for 7 days or with AICAR intraperitoneally at 100 mg/kg 24 hours before death. Retinal protein levels of phosphorylated and total AMPK, phosphorylated nuclear factor (NF)-κB p65, intercellular adhesion molecule (ICAM)-1, and vascular endothelial growth factor (VEGF) were evaluated by Western blot analysis or enzyme-linked immunosorbent assay. Retinal activity of sirtuin (SIRT)1 was measured by deacetylase fluorometric assay. Leukocyte adhesion to the retinal vasculature was examined with a concanavalin A lectin perfusion-labeling technique. Results. Induction of diabetes in mice led to retinal AMPK dephosphorylation, which was significantly reversed by either resveratrol or AICAR. Either resveratrol or AICAR significantly reversed SIRT1 deactivation and NF-κB phosphorylation, both of which were induced in the diabetic retina. Administration of resveratrol to diabetic mice significantly reduced diabetes-induced retinal leukocyte adhesion, together with retinal expression of ICAM-1 and VEGF. Conclusions. The present findings reveal that diabetes-induced retinal inflammation stems from downregulation of the AMPK pathway, leading subsequently to SIRT1 deactivation and NF-κB activation. The data also suggest the potential use of the AMPK activator resveratrol as a therapeutic agent for diabetic retinopathy. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Effect of controlled adverse chamber environment exposure on tear functions in silicon hydrogel and hydrogel soft contact lens wearers

Kojima Takashi, Matsumoto Yukihiro, Ibrahim Osama M A, Wakamatsu Tais Hitomi, Uchino Miki, Fukagawa Kazumi, Ogawa Junko, Dogru Murat, Negishi Kazuno, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 12 ) 8811 - 8817 2011.11

ISSN 0146-0404

Purpose. To prospectively evaluate the effect of controlled adverse chamber environment (CACE) exposure on tear function, including tear osmolarity, in subjects wearing narafilcon A versus those wearing etafilcon A soft contact lens (SCL). Methods. Thirty-one healthy subjects with no history of contact lens wear (13 women, 18 men; average age, 30.5 ± 6.5 years) were randomly divided into age- and sex-matched groups (15 subjects wearing narafilcon A SCL; 16 subjects wearing etafilcon A SCL) and entered a CACE for 20 minutes. All subjects underwent tear osmolarity, tear evaporation rate, strip meniscometry, tear film breakup time, fluorescein vital staining, and functional visual acuity measurement before and after exposure to the controlled adverse chamber. Results. The mean blink rate increased with significant deteriorations in the mean symptom VAS scores, mean tear osmolarity, tear evaporation rate, strip meniscometry score, and tear stability with CACE exposure along with a decrease in visual maintenance ratio in functional visual acuity testing in etafilcon A wearers. The mean symptom VAS scores, mean tear evaporation rate, tear stability, blink rates, and visual maintenance ratios did not change significantly in narafilcon A wearers after CACE exposure. Conclusions. This study suggested marked tear instability, higher tear osmolarity, and increased tear evaporation with marked dry eye and visual symptomatology in nonadapted hydrogel SCL wearers, suggesting that silicone hydrogel SCLs may be suitable for persons who live and work in cool, low-humidity, and windy environments, as tested in this study. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Regulatory mechanisms for the production of BAFF and IL-6 are impaired in monocytes of patients of primary Sjogren's syndrome

Yoshimoto Keiko, Tanaka Maiko, Kojima Masako, Setoyama Yumiko, Kameda Hideto, Suzuki Katsuya, Tsuzaka Kensei, Ogawa Yoko, Tsubota Kazuo, Abe Tohru, Takeuchi Tsutomu

Arthritis Research & Therapy 2011.10

ISSN 1478-6354

Introduction: In this study, we investigated possible aberrations of monocytes from patients with primary Sjogren's syndrome (pSS). We focused on BAFF (B cell activating factor of the TNF family) and IL-6, because they are both produced by monocytes and known to be involved in the pathogenesis of pSS. Methods: Peripheral monocytes were prepared from both pSS patients and normal individuals. The cells were stimulated in vitro with IFN-gamma, and the amounts of IL-6 and sBAFF produced by the cells were quantitated. The effect of sBAFF itself on the production of IL-6 was also studied. To investigate the response of pSS monocytes to these stimuli, the expression levels of the genes encoding BAFF receptors and IL-6-regulating transcription factors were quantitated. Results: Peripheral pSS monocytes produced significantly higher amounts of sBAFF and IL-6 compared to normal monocytes, even in the absence of stimulation. The production of these cytokines was significantly increased upon stimulation with IFN-gamma. The elevated production of IL-6 was significantly suppressed by an anti-BAFF antibody. In addition, stimulation of pSS monocytes with sBAFF induced a significant increase in IL-6 production. Moreover, the expression levels of a BAFF receptor and transcription factors regulating IL-6 were significantly elevated in pSS monocytes compared to normal monocytes. Conclusions: The present study suggests that the mechanisms for the production of sBAFF and IL-6 are impaired in pSS monocytes. Our research implies that this impairment is ascribed to abnormally overexpressed IL-6-regulating transcription factors and a BAFF receptor. These abnormalities may underlie the development of pSS.

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Acute angle-closure glaucoma in a case each of cicatricial retinopathy of prematurity and persistent hyperplastic primary vitreous

Katsumura Takahiro, Tanabe Sachiko, Ozeki Naoki, Yuki Kenya, Tanino Tomihiko, Shiba Daisuke, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 65 ( 10 ) 1631 - 1634 2011.10

ISSN 0370-5579

Purpose: To report acute angle-closure glaucoma in an eye with cicatricial retinopathy of prematurity and in another with persistent hyperplastic primary vitreous (PHPV). Cases: One was a 28-year-old female with PHPV in both eyes. Her right eye developed acute angle-closure glaucoma with intraocular pressure (IOP) of 48 mmHg. The chamber angle was closed in all the circumference. Lens extraction resulted in increased anterior chamber depth with decreased IOP. The other case was a 20-year-old male with cicatricial retinopathy of prematurity. He had received peripheral iridectomy in the right eye due to elevated IOP at the age of 14. IOP in the right eye rose to 34 mmHg after anterior dislocation of the lens. Lens extraction induced increased anterior chamber depth with decreased IOP. Conclusion: These cases illustrate that lens extraction may be effective for pupillary block after laser iridotomy or peripheral iridectomy has failed.

Fate of corneal epithelial cells separated from limbus in vivo

Kawakita Tetsuya, Higa Kazunari, Shimmura Shigeto, Tomita Machiko, Tsubota Kazuo, Shimazaki Jun

Investigative Ophthalmology and Visual Science 52 ( 11 ) 8132 - 8137 2011.10

ISSN 0146-0404

Purpose. To characterize corneal epithelial cells separated from limbus in vivo by transplantation of a stainless steel ring with or without creating a defect inside the ring. Methods. A stainless steel ring (diameter, 8 mm; width, 300 μm; depth, 250 μm) was transplanted into rabbit corneal stroma using 10-0 nylon interrupted suture after cutting to a 250 μm depth by corneal vacuum trephine (diameter, 8.0 mm). Epithelial cells were removed inside the ring, and re-epithelization was evaluated after 1 week. Hematoxylin staining and immunostaining against p63, Ki67, and cytokeratin 3 were performed for phenotypic analysis of corneal epithelia. A corneal epithelial defect was centrally created inside the ring (4, 5, and 6 mm diameter) after transplantation. When re-epithelization was achieved, a central epithelial defect was continuously created until cells were exhausted within the ring. The number of created defects was also analyzed to assess the potential of re-epithelialization. Results. Ring-transplanted corneal stroma showed few signs of inflammation, and when epithelium was totally removed from inside the ring, complete epithelial defects were persistent for ≥ 1 month. Corneal sensation was significantly decreased in corneas with the ring (P < 0.05). Immunostaining demonstrated similar expression patterns for p63, Ki67, and cytokeratin3 as the controls. When rings were transplanted into the intact cornea, inside epithelia prevented epithelial defects in vivo for ≤ 6 months. Conclusions. Transient-amplifying cells might maintain homeostasis for >1 month when separated from their limbus in vivo. This model will be useful for future stem cell research or wound healing models. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Concluding my term as president with great appreciation and satisfaction 16th annual meeting of the Kyoto Cornea club, december 3-4, 2010

Tsubota Kazuo, Inoue Yoshitsugu, Oshika Tetsuro

Cornea 30 ( 10 SUPPL. 1 ) 2011.10

ISSN 0277-3740

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The comparison of solitary topical micafungin or fluconazole application in the treatment of Candida fungal keratitis

Matsumoto Yukihiro, Murat Dogru, Kojima Takashi, Shimazaki Jun, Tsubota Kazuo

British Journal of Ophthalmology 95 ( 10 ) 1406 - 1409 2011.10

ISSN 0007-1161

Aim: To compare and evaluate the efficacy of topical 0.1% micafungin (MCFG) and topical 0.2% fluconazole (FCZ) in the treatment of Candida fungal keratitis. Methods: Twenty-nine eyes of 29 patients who were diagnosed as having Candida fungal keratitis, proven by corneal culture isolates, were investigated in this prospective study. Patients were divided into a MCFG treatment group (12 eyes) and an FCZ treatment group (17 eyes). Age, gender, initial status of ulcer (ulcer size and degree of injection), initial and final best-corrected visual acuity (BCVA), healing periods, final status of cornea and recurrences in each group were studied and compared. Results: There were no significant differences in relation to age, gender, ulcer size and degree of injection before treatment between the two groups. There were also no significant differences in the healing periods until complete epithelialisation (MCFG treatment group (41.3±38.0 days); FCZ treatment group (34.4±37.7 days)), change in BCVA, corneal clarity/ opacification, perforation and recurrence status at the final examination between the two groups. Conclusion: The efficacy of 0.1% MCFG eye-drops appears to be comparable with that of 0.2% FCZ eye-drops in the treatment of Candida fungal keratitis.

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Effect of calorie restriction on change in lacrimal gland with age

Kawashima Motoko, Tsubota Kazuo

Cornea 30 ( 10 SUPPL. 1 ) 2011.10

ISSN 0277-3740

Rapid growth in the number of dry eye syndrome patients has been observed worldwide. Increased life span is considered to be at least part of the reason for this because there is a close association between aging and decreased secretory function in the lacrimal gland. Recent advances have led to a new way of thinking about intervention in the aging process. Calorie restriction (CR) is believed to retard functional decline in various organs with age. We hypothesized that CR could prevent age-related lacrimal dysfunction. In this paper, we begin by reviewing the epidemiology of dry eye syndrome and explaining how aging affects lacrimal gland function. We then introduce a possible therapeutic approach to this problem, intervention by means of CR. Finally, suggestions are made for further prospective research that would elucidate the molecular mechanisms by which CR maintains lacrimal gland function. © 2011 by Lippincott Williams & Wilkins.

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Mathematical projection model of visual loss due to fuchs corneal dystrophy

Hatou Shin, Shimmura Shigeto, Shimazaki Jun, Usui Tomohiko, Amano Shiro, Yokogawa Hideaki, Kobayashi Akira, Zheng Xiaodong, Shiraishi Atsushi, Ohashi Yuichi, Inatomi Tsutomu, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 11 ) 7888 - 7893 2011.10

ISSN 0146-0404

Purpose. To devise a mathematical disease classification model for eyes with primary guttata cornea, on the bases of endothelial loss trajectory and probability of advanced disease. Methods. A series of 1971 patients (3281 eyes), some with and some without guttata corneas, undergoing specular microscopy were retrospectively reviewed. The eyes were classified into four stages; stage 0, without guttae; 1, guttata cornea without edema; 2, mild Fuchs' corneal dystrophy (FCD); and 3, severe FCD, according to clinical records, and patient age and corneal endothelial cell density (ECD) were plotted. Nonparametric density smoothing was used to create a contour map, and a best-fit curve for ECD loss was calculated. The relation between ECD decrease rate and the stages were evaluated. Results. Endothelial decrease rate in stage 0 was 0.44%/year, which was compatible with that of normal eyes reported in previous studies. Decrease rates of stages 1, 2, and 3 were 0.81%, 2.65%, and 3.08%/ year, respectively. The age-ECD loss curves of 1.40%/year (ECO _1.4) and 2.00%/year (ECO _2.0) further divided stage 1 into three subgroups; stage 1a, asymptomatic guttata cornea; 1b, borderline guttata cornea; and 1c, pre-FCD. The ECO _2.0 cutoff line differentiated eyes with FCD from those without edema with a sensitivity and specificity of >90%. Stage 1c eyes were below ECO _2.0 and had a decrease rate as high as FCD. Conclusions. This mathematical model can be used to predict the prognosis of patients with primary guttata cornea. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Morphologic evaluation of meibomian glands in chronic graftversus- host disease using in vivo laser confocal microscopy

Ban Yumiko, Ogawa Yoko, Ibrahim Osama M A, Tatematsu Yukako, Kamoi Mizuka, Uchino Miki, Yaguchi Saori, Dogru Murat, Tsubota Kazuo

Molecular Vision 17 2533 - 2543 2011.09

Purpose: To evaluate the morphological changes of the meibomian glands (MGs) using in vivo laser confocal microscopy (CM) in dry eye (DE) patients with chronic graft-versus-host disease (cGVHD). Methods: Seventeen eyes from 9 patients with a diagnosis of DE associated with cGVHD (DE/cGVHD group; 6 males, 3 females; median 50.5 years) and 16 eyes of 8 hematopoietic stem cell transplantation (HSCT) recipients without DE (non-DE/non-cGVHD group; 5 males, 3 females; median 47.0 years) were enrolled. CM was used to investigate the MG and MG acinar unit density (MGAUD), MG acinar longest diameter (MGALD), MG acinar shortest diameter (MGASD), and the fibrosis grade. Clinical findings of the lid margin were obtained. Tear dynamics, ocular surface vital staining, meibography, and MG expressibility were also examined. Data were compared between the 2 groups using the unpaired t and Mann-Whitney tests. Results: The mean MGAUD value was significantly lower in the DE/cGVHD group than in the non-DE/non-cGVHD group (p=0.01, 57.8±38.3 glands/mm ², 88.8±26.6 glands/mm ², respectively), and the mean MGALD and MGASD were significantly shorter in the DE/cGVHD group than in the non-DE/non-cGVHD group (p=0.0018, 37.3±24.4 μm and 60.4±11.8 μm, p=0.0106, 17.7±11.8 μm and 26.6±6.03 μm, respectively). The mean fibrosis grade was significantly higher in the DE/cGVHD group than the non-DE/non-cGVHD group (p<0.0001, 1.39±0.71 grade, 0.06±0.25 grade, respectively). Clinical findings in the lid margin, tear dynamics, and ocular surface findings were significantly worse in the DE/cGVHD group than in the non-DE/non-cGVHD group. Conclusions: CM clearly depicted the morphological changes of the MG in the DE/cGVHD group, and revealed the severity of the meibomian gland dysfunction. Patients with severe DE after HSCT showed atrophic MG and excessive fibrosis. © 2011 Molecular Vision.

IL-6 induction in desiccated corneal epithelium in vitro and in vivo

Higuchi Akihiro, Kawakita Tetsuya, Tsubota Kazuo

Molecular Vision 17 2400 - 2406 2011.09

Purpose: To investigate the effect of desiccation on secretion of inflammatory cytokines in corneal epithelial cells and in the rat desiccation model. Methods: A human corneal epithelial cell line (CEPI) was grown in keratinocyte growth medium 2 (KGM2) to approximately 80% confluence. The medium was aspirated and dishes were left for 0 to 30 min with the cover left open to dry the cells (short-term desiccation). After desiccation, KGM2 was added to the dishes and collected from the dishes 15 min later to measure the concentrations of cytokines in the medium by sandwich enzyme immunoassay (ELISA). Viability of the cells was estimated with alamer blue. To study the effect of long-term desiccation, cultivated cells on transwells were used. After dessiccation for up to 8 h, the viability of the cells and levels of cytokines in the culture medium were examined. The expression of cytokines in the cornea of the dry eye model rat was measured by real-time PCR. Results: Short-term dessication of CEPI cells significantly increased the interleukin (IL)-6 level and slightly increased the tumor necrosis factor (TNF)-α level. Anti-IL-6 antibody partially suppressed cell death caused by desiccation. Upon long-term desiccation, IL-6 and IL-8 levels were increased. In the dry eye model rats, the IL-6 mRNA level in the cornea significantly increased, whereas TNF-α mRNA level slightly increased. Conclusions: Desiccation induced IL-6 expression in corneal epithelial cells, suggesting that IL-6 participates in desiccation-induced cell death. © 2011 Molecular Vision.

The relation of functional visual acuity measurement methodology to tear functions and ocular surface status

Kaido Minako, Ishida Reiko, Dogru Murat, Tsubota Kazuo

Japanese Journal of Ophthalmology 55 ( 5 ) 451 - 459 2011.09

ISSN 0021-5155

Purpose: To investigate the relation of functional visual acuity (FVA) measurements with dry eye test parameters and to compare the testing methods with and without blink suppression and anesthetic instillation. Design: A prospective comparative case series. Methods: Thirty right eyes of 30 dry eye patients and 25 right eyes of 25 normal subjects seen at Keio University School of Medicine, Department of Ophthalmology were studied. FVA testing was performed using a FVA measurement system with two different approaches, one in which measurements were made under natural blinking conditions without topical anesthesia (FVA-N) and the other in which the measurements were made under the blink suppression condition with topical anesthetic eye drops (FVA-BS). Tear function examinations, such as the Schirmer test, tear film break-up time, and fluorescein and Rose Bengal vital staining as ocular surface evaluation, were performed. Results: The mean logMAR FVA-N scores and logMAR Landolt visual acuity scores were significantly lower in the dry eye subjects than in the healthy controls (p < 0.05), while there were no statistical differences between the logMAR FVA-BS scores of the dry eye subjects and those of the healthy controls. There was a significant correlation between the logMAR Landolt visual acuities and the logMAR FVA-N and logMAR FVA-BS scores. The FVA-N scores correlated significantly with tear quantities, tear stability and, especially, the ocular surface vital staining scores. Conclusions: FVA measurements performed under natural blinking significantly reflected the tear functions and ocular surface status of the eye and would appear to be a reliable method of FVA testing. FVA measurement is also an accurate predictor of dry eye status. © 2011 Japanese Ophthalmological Society.

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Evaluation of tear stability after surgery for conjunctivochalasis

Hara Shuya, Kojima Takashi, Ishida Reiko, Goto Eiki, Matsumoto Yukihiro, Kaido Minako, Shimazaki Jun, Dogru Murat, Tsubota Kazuo

Optometry and Vision Science 88 ( 9 ) 1112 - 1118 2011.09

ISSN 1040-5488

Purpose.: To evaluate the efficacy of surgical treatment for conjunctivochalasis using the Tear Film Stability Analysis System. Methods.: Fifteen eyes of 15 patients (74.5 ± 4.8 years; 14 females and 1 male) who had surgical reconstruction of the tear meniscus for conjunctivochalasis underwent evaluation with a symptom questionnaire, Schirmer I test, fluorescein tear film break-up time (FBUT), and break-up time index (BUI) measurement by Tear Film Stability Analysis System examinations that were performed preoperatively and 3 months after the surgery. Changes of these tear parameters were statistically analyzed using the paired t test. Results.: After the surgery, the mean FBUT and BUI values significantly improved from 5.7 ± 3.2 s and 42.8 ± 18.7, respectively, to 8.4 ± 2.5 s and 60.7 ± 16.5, respectively (FBUT, p = 0.0438; BUI, p = 0.0036). The mean Schirmer test value showed no significant changes before and after surgery (p = 0.9064). Fourteen eyes (93.3%) had improvement of ocular symptoms after surgery. One eye (6.7%) with no improvement of ocular symptoms, FBUT, and BUI after surgery appeared to have aqueous tear deficiency. Conclusions.: Surgical treatment for conjunctivochalasis was associated with improvement of tear stability and clinical dry eye symptoms. Copyright © 2011 American Academy of Optometry.

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Prevention of cytarabine-induced kerato-conjunctivitis by eye rinse in patients receiving high-dose cytarabine and total body irradiation as a conditioning for hematopoietic stem cell transplantation

Mori Takehiko, Kato Jun, Yamane Akiko, Aisa Yoshinobu, Kawata Yusuke, Ichimura Mayu, Tsuneyama Chiaki, Yashima Tomoko, Ogawa Yoko, Tsubota Kazuo, Shigematsu Naoyuki, Shimizu Takayuki, Okamoto Shinichiro

International Journal of Hematology 94 ( 3 ) 261 - 265 2011.09

ISSN 0925-5710

We previously reported a high incidence of kerato-conjunctivitis in patients receiving high-dose cytarabine following total body irradiation (TBI) as a conditioning for hematopoietic stem cell transplantation (HSCT) even on prophylaxis with topical corticosteroid. This study aimed to evaluate whether addition of eye rinse, which was designed to remove cytarabine from ocular surface, further reduces the incidence of kerato-conjunctivitis in the same setting. Seventy-six patients receiving cytarabine at a dose of 3 g/m ² every 12 h for 4 days after receiving TBI (12 Gy) as conditioning for HSCT were evaluated. All patients received betamethasone sodium phosphate eye drops. Twenty-three patients were further instructed to rinse their eyes with sterile saline every 10-15 min during and for two additional hours after the completion of each cytarabine infusion. Among 23 patients with eye rinse, Grades 2-3 and 1-3 kerato-conjunctivitis were observed in 4 (17.4%) and 5 patients (21.7%), respectively. These incidences were significantly lower than those [35 (66.0%) and 41 (77.4%)] observed in 53 patients without eye rinse (P<0.001 and P<0.00001, respectively). These results strongly suggest that eye rinse effectively reduces the incidence and severity of cytarabine-induced kerato-conjunctivitis in HSCT recipients who receive high-dose cytarabine following TBI.

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Long-term outcome of cultivated oral mucosal epithelial sheet transplantation in treatment of total limbal stem cell deficiency

Satake Yoshiyuki, Higa Kazunari, Tsubota Kazuo, Shimazaki Jun

Ophthalmology 118 ( 8 ) 1524 - 1530 2011.08

ISSN 0161-6420

Purpose: To evaluate the long-term outcome of cultivated oral mucosal epithelial transplantation (COMET) in treatment of eyes with total limbal stem cell deficiency. Design: Noncomparative, retrospective, interventional case series. Participants: Forty eyes in 36 patients with total limbal stem cell deficiency (Stevens-Johnson syndrome in 12 eyes, chemical or thermal burns in 11 eyes, ocular cicatricial pemphigoid [OCP] in 9 eyes, pseudo-OCP in 7 eyes, and gelatinous drop-like dystrophy in 1 eye) were treated at the Department of Ophthalmology, Tokyo Dental College, Chiba, Japan. Intervention: Cultivated autologous oral mucosal epithelial sheets were transplanted onto the ocular surface in eyes with total limbal stem cell deficiency. Main Outcome Measures: Reconstruction of a stable ocular surface with a clear appearance and no epithelial defects, reduction in fibrovascular tissue invasion of corneal surface, a functional fornix, change in visual acuity, and postoperative complications. Results: The mean follow-up period was 25.5 months (range, 654.9 months). Kaplan-Meier analysis of a corneal surface stability revealed an early decline in transplanted oral mucosal epithelial stability over the first 6 months, remaining comparatively stable thereafter (1 year, 64.8%; 2 years, 59.0%; and 3 years, 53.1%). Postoperative persistent epithelial failure developed within the first 3 months in 9 eyes. Early epithelial failure was associated closely with preoperative corneal defects. Gradual fibrovascular tissue invasion of the corneal surface was observed in 8 eyes and was marked in cases of OCP. Survival of a functional fornix decreased progressively until approximately 6 months. Postoperative visual acuity seemed to be related to the presence of corneal opacity. Complications included stromal melting or perforation in 8 eyes, infectious keratitis in 2 eyes, glaucoma in 8 eyes, and recurrence of herpetic keratitis in 1 eye. Corneal melting or perforation and infectious keratitis were associated closely with persistent epithelial defects after COMET. Conclusions: The transplantation of cultivated oral mucosal epithelial sheets offers a viable and safe alternative in the reconstruction of a stable ocular surface. Epithelialization of the corneal surface is very important not only in obtaining a satisfactory long-term outcome, but also in achieving a lower incidence of complications. Financial Disclosure(s): The author(s) have no proprietary or commercial interest in any materials discussed in this article. © 2011 American Academy of Ophthalmology.

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The contribution of the posterior surface to the corneal aberrations in eyes after keratoplasty

Yamaguch Takefumi, Ohnuma Kazuhiko, Tomida Daisuke, Konomi Kenji, Satake Yoshiyuki, Negishi Kazuno, Tsubota Kazuo, Shimazaki Jun

Investigative Ophthalmology and Visual Science 52 ( 9 ) 6222 - 6229 2011.08

ISSN 0146-0404

Purpose. To investigate the contribution of posterior corneal surfaces to higher-order aberrations (HOAs) of the cornea, optical quality, and visual acuity after keratoplasty. METHODS. Corneal topography of anterior and posterior surfaces and pachymetry were conducted using anterior segment optical coherence tomography (AS-OCT) in 40 eyes (10 eyes after penetrating keratoplasty [PK], 10 eyes after deep anterior lamellar keratoplasty [DALK], 10 eyes after Descemet's stripping automated endothelial keratoplasty [DSAEK], and 10 normal eyes). Anterior, posterior, and total corneal HOAs were calculated using ray-tracing and decomposition into Zernike polynomials and were evaluated as root mean square values. Modulation transfer functions (MTFs) were also evaluated. RESULTS. Topography maps of the anterior and posterior surfaces showed reverse patterns in the normal, PK, and DALK eyes, but not in DSAEK eyes. In the normal, PK, and DALK eyes, the total corneal HOAs were significantly smaller (~10%) than were the HOAs of the anterior surface (P<0.01), whereas there was no significant difference between total and anterior HOAs in the DSAEK eyes (P=0.483). In the normal, PK, and DALK eyes, the MTFs of the total cornea were slightly better than those of the anterior surface. In the DSAEK eyes, the MTFs of the total cornea were lower than those of the anterior surface. Visual acuity was significantly correlated with total and anterior surface HOAs (P<0.05). CONCLUSIONS. Posterior surfaces compensate for anterior aberrations in normal, PK, and DALK eyes. In DSAEK eyes, the posterior surface increased total corneal HOAs and had a negative influence on MTFs. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Repositioning and scleral fixation of subluxated lenses using a T-shaped capsule stabilization hook

Yaguchi Saori, Yaguchi Shigeo, Asano Yasuhiko, Kozawa Tadahiko, Miyawaki Takaya, Negishi Kazuno, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 37 ( 8 ) 1386 - 1393 2011.08

ISSN 0886-3350

We describe a T-shaped capsule stabilization hook (modified capsule expander [M-CE]) used for repositioning and scleral fixation of the lens capsule of subluxated lenses. The 5-0 polypropylene device is flexible and attached to a curved needle. The contact portion is bent at 1.25 mm, and the end bifurcates in a T configuration to form a 3.75 mm footpad from which the capsular bag can be suspended. Modified capsule expanders were implanted in 4 eyes of 4 patients with subluxated cataractous lenses and provided excellent support and centration of the intraocular lens (IOL)-capsular bag complex. The IOLs remained well centered and stable. The corrected distance visual acuity improved to at least 20/20 in all patients after surgery. Thus, M-CEs were effective in fixating the lens capsule to the sclera in patients with significant zonular weakness. Financial Disclosure: Saori Yaguchi, Y. Asano, T. Kozawa, T. Miyawaki, K. Negishi, and K. Tsubota have no financial or proprietary interest in any material or method mentioned. Additional disclosure is found in the footnotes. © 2011 ASCRS and ESCRS.

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Resveratrol prevents the development of abdominal aortic aneurysm through attenuation of inflammation, oxidative stress, and neovascularization

Kaneko Hidehiro, Anzai Toshihisa, Morisawa Maho, Kohno Takashi, Nagai Toshiyuki, Anzai Atsushi, Takahashi Toshiyuki, Shimoda Masayuki, Sasaki Aya, Maekawa Yuichiro, Yoshimura Koichi, Aoki Hiroki, Tsubota Kazuo, Yoshikawa Tsutomu, Okada Yasunori, Ogawa Satoshi, Fukuda Keiichi

Atherosclerosis 217 ( 2 ) 350 - 357 2011.08

ISSN 0021-9150

Objective: We sought to examine the effect of resveratrol (3,4',5-trihydroxy-trans-stilbene), a plant-derived polyphenolic compound, on the development of abdominal aortic aneurysm (AAA). Methods: AAA was induced in mice by periaortic application of CaCl ₂. NaCl (0.9%)-applied mice were used as a sham group. Mice were treated with intraperitoneal injection of PBS (Sham/CON, AAA/CON, n=30 for each) or resveratrol (100mg/kg/day) (AAA/RSVT, n=30). Six weeks after the operation, aortic tissue was excised for further examinations. Results: Aortic diameter was enlarged in AAA/CON compared with Sham/CON. Resveratrol treatment reduced the aneurysm size and inflammatory cell infiltration in the aortic wall compared with AAA/CON. Elastica Van Gieson staining showed destruction of the wavy morphology of the elastic lamellae in AAA/CON, while it was preserved in AAA/RSVT. The increased mRNA expression of monocyte chemotactic protein-1, tumor necrosis factor-α, intercellular adhesion molecule-1, CD68, vascular endothelial growth factor-A, p47, glutathione peroxidase (GPX)1 and GPX3 were attenuated by resveratrol treatment (all p<0.05). Administration of resveratrol decreased protein expression of phospho-p65 in AAA. The increased 8-hydroxy-2'-deoxyguanosine-positive cell count and 4-hydroxy-2-nonenal-positive cell count in AAA were also reduced by resveratrol treatment. Zymographic activity of matrix metalloproteinase (MMP)-9 and MMP-2 was lower in AAA/RSVT compared with AAA/CON (both p<0.05). Compared with AAA/CON, Mac-2 ⁺ macrophages and CD31 ⁺ vessels in the aortic wall were decreased in AAA/RSVT (both p<0.05). Conclusion: Treatment with resveratrol in mice prevented the development of CaCl ₂-induced AAA, in association with reduced inflammation, oxidative stress, neoangiogenesis, and extracellular matrix disruption. These findings suggest therapeutic potential of resveratrol for AAA. © 2011 Elsevier Ireland Ltd.

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The application of in vivo confocal scanning laser microscopy in the diagnosis and evaluation of treatment responses in mooren's ulcer

Hatou Shin, Dogru Murat, brahim Osama M A, Wakamatsu Tais, Sato Enrique Adan, Shimmura Shigeto, Negishi Kazuno, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 9 ) 6680 - 6689 2011.08

ISSN 0146-0404

Purpose. To evaluate the extent of corneal inflammation and the response to treatment in patients with Mooren's ulcer, by in vivo confocal microscopy (IVCM). Methods. Twenty-two eyes of 15 patients with Mooren's ulcer were enrolled in this prospective study. All subjects underwent routine ophthalmic examinations, IVCM, and conjunctival histopathologic examination of specimens in patients undergoing conjunctival excision. Eyes with active ulcer were treated with topical corticosteroids and additional therapy, depending on the signs and symptoms. Eyes in remission continued to receive the previous treatment protocols. The relation between the severity of ulcer and inflammation status as assessed by IVCM was also studied. The endpoints were inflammatory cell density (ICD), counted by IVCM, and the extent of the limbal arc involved with ulcers. Results. Ten eyes had active corneal ulcers, and 12 eyes had been in remission for the past year. The mean ICD of eyes with active ulcers before treatment was 2092.7 ± 1538.6 cells/mm2 (range, 835.3-7832.7; 95% CI, 3232.5-952.9). Nine of the eyes improved at 8 weeks, with a decrease in ICD to 249.1 ± 109.0 cells/mm2 (range, 100.3-595.3; 95% CI, 329.8-168.3). One eye had corneal perforation, and ICD immediately before perforation was 1677.6 ± 247.6 cells/mm2. The mean ICD of 12 eyes in remission was 357.5 ± 266.8 cells/mm2 (range, 12.7-1127.0; 95% CI, 555.2-159.8). The correlation of the ICD and the extent of limbal involvement with ulcers was strong (R2 = 0.8119). Conclusions. ICD evaluated by IVCM is a useful and promising parameter for evaluation of the degree of inflammation in eyes with Mooren's ulcer and for assessment of response to treatment. © Association for Research in Vision and Ophthalmology.

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Prospective, randomized study of the efficacy of systemic cyclosporine in high-risk corneal transplantation

Shimazaki Jun, Den Seika, Omoto Masahiro, Satake Yoshiyuki, Shimmura Shigeto, Tsubota Kazuo

American Journal of Ophthalmology 152 ( 1 ) 33 - 39 2011.07

ISSN 0002-9394

Purpose: Immunologic rejection remains a major cause of graft failure in high-risk corneal transplantation. This study was conducted to elucidate the efficacy and safety of systemic cyclosporine (CsA) in high-risk corneal transplantation. Design: Prospective, randomized, open-labeled clinical trial with a parallel-group study. Methods: Patients underwent high-risk corneal transplantation at the Department of Ophthalmology, Tokyo Dental College, Chiba, Japan. High-risk was defined as corneal neovascularization in more than 1 quadrant or a history of corneal grafting. Patients were assigned to either a systemic CsA group or a control group. Administration of CsA was continued for at least 6 months with blood CsA concentration 2 hours after administration of approximately 800 ng/mL, unless undesirable side effects developed. The main outcome measures were graft clarity, endothelial rejection, and local and systemic complications. Results: Forty patients were enrolled and 39 (18 men, 21 women; mean age, 67.4 ± 11.9 years) were analyzed. In the CsA group, CsA was discontinued within 6 months in 7 patients because of side effects. With a mean follow-up of 42.7 months, endothelial rejection developed in 6 and 2 eyes in the CsA and control groups, respectively. No differences were observed in the rates of graft clarity loss between the 2 groups (P = .16, Kaplan-Meier analysis). Conclusions: No positive effect of systemic CsA administration for suppressing rejection in high-risk corneal transplantation was observed. With a relatively high incidence of systemic side effects, the results suggest that this protocol should not be recommended for corneal transplant recipients, especially those of advanced age. © 2011 Elsevier Inc.

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Donor-recipient gender difference affects severity of dry eye after hematopoietic stem cell transplantation

Kamoi M., Ogawa Y., Uchino M., Tatematsu Y., Mori T., Okamoto S., Tsubota K.

Eye 25 ( 7 ) 860 - 865 2011.07

ISSN 0950-222X

Purpose To determine whether the incidence rate and severity of dry eye after hematopoietic stem cell transplantation varies with donor vsrecipient gender. Methods We limited this study to patients received bone marrow transplantation (BMT). In all, 172 patients received BMT at Keio University School of Medicine between January 2000 and May 2007. Of them, 136 recipients who survived at least 70 days were studied prospectively. We classified the 136 patients according to the gender of the donor and the recipient (group I: female to female; group II: male to male; group III: male to female; group IV: female to male). The incidence and severity of chronic graft-vs-host disease-associated dry eye were determined for each group. The donor gender was masked when we assessed dry eye and calculate the incidence. Results The incidence of dry eye was 47.4% for group I, 37.5% for group II, 58.6% for group III, and 42.9% for group IV. The percentage of patients with severe dry eye was 44.4, 50.0, 35.3, and 77.8% respectively. There was a significant difference between the percent severe dry eye/total dry eye incidences in groups III and IV (P=0.0375) (odds ratio, 7.6; 95% confidence interval, 1.00-101.01).Conclusions Close attention must be paid to the development of dry eye in cases of female to male BMTs, because the ratio of severe/total dry eye is more common in cases of female to male BMTs than in other gender combination. © 2011 Macmillan Publishers Limited All rights reserved.

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Incidence of increased intraocular pressure after subtenon injection of triamcinolone acetonide

Kawamura Ryosuke, Inoue Makoto, Shinoda Hajime, Shinoda Kei, Itoh Yuji, Ishida Susumu, Tsubota Kazuo

Journal of Ocular Pharmacology and Therapeutics 27 ( 3 ) 299 - 304 2011.06

ISSN 1080-7683

Purpose: To determine the incidence of eyes that have an increase in the intraocular pressure (IOP) after subtenon injections of triamcinolone acetonide (TA). Methods: The medical records of 147 patients treated with single or multiple subtenon injections of TA (10 mg) were reviewed. The incidence of an IOP elevation (ΔIOP) ≥5 mmHg or an IOP of >21 mmHg was determined. The peak ΔIOP, defined as the difference in the IOP at the peak to the baseline IOP, was also evaluated. Multivariate analyses were used to evaluate the relation between the IOP elevation and the age, gender, refractive error, and lens status. Results: A ΔIOP ≥5 mmHg was found in 75 eyes (46%), and an IOP >21 mmHg was found in 48 eyes (30%) after a single injection of TA. The IOP increased within 3 months in 39 eyes (81%) and after 4 months in 9 eyes (19%). The IOP began to increase significantly from 2 weeks up to 5 months (P<0.05) and returned to the baseline IOP in 10 months. The incidence of ΔIOP ≥5 mmHg or an IOP of >21 mmHg after multiple subtenon injections of TA was significantly higher than after a single injection (62%; P=0.027, 47%; P=0.013, respectively). The incidence of IOP >21 mmHg and the peak ΔIOP were significantly related with younger age (P=0.002, P=0.021, Forward stepwise regression analysis). A weak but significant negative correlation was found between the peak ΔIOP and the age (r=-0.216, P=0.006, Pearson's correlation coefficient test), and the peak ΔIOP and the refractive error (r=-0.198; P=0.018). Conclusion: Repeated injections of TAs and injection of younger patients or myopic eyes increase the incidence of an IOP elevation. © 2011, Mary Ann Liebert, Inc.

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Topography-guided conductive keratoplasty for keratoconus

Kato Naoko, Tsubota Kazuo

American Journal of Ophthalmology 151 ( 6 ) 2011.06

ISSN 0002-9394

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The association between primary open-angle glaucoma and motor vehicle collisions

Tanabe Sachiko, Yuki Kenya, Ozeki Naoki, Shiba Daisuke, Abe Takayuki, Kouyama Keisuke, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 7 ) 4177 - 4181 2011.06

ISSN 0146-0404

Purpose. To investigate and compare the prevalence of motor vehicle collisions (MVCs) in individuals with or without primary open-angle glaucoma (POAG). Methods. A total of 265 subjects were consecutively enrolled: 121 (79 men, 42 women; age, 62.1 ± 8.0 years) with POAG; and 144 (95 men, 49 women; age, 61.2 ± 7.9 years) who were free of ocular disease. Participants answered a questionnaire on MVC experience during the previous 10 years, past driving experience, and daily driving habits. The POAG group was subdivided into three groups according to disease severity (mild, moderate, or severe), to assess the relationship between POAG severity and MVC. Results. A statistically significant association between POAG severity and MVC frequency was observed; 3.5% of the controls, 0.0% of the mild POAG group, 3.9% of the moderate POAG group, and 25.0% of the severe POAG group had experienced MVCs (P = 0.007, Cochran-Armitage trend test). The severe POAG group had experienced a much higher frequency of MVCs during the surveyed period than had the control group (P < 0.010; Fisher's exact test). Logistic regression analyses to account for confounding factors (age, presence of diabetes mellitus, driving history, time spent driving per day, and best corrected visual acuity in the better or worse eye) produced consistent results. Conclusions. Advanced POAG with marked visual field defects may be a risk factor for MVCs. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Retinal ganglion cell loss in superoxide dismutase 1 deficiency

Yuki Kenya, Ozawa Yoko, Yoshida Tetsu, Kurihara Toshihide, Hirasawa Manabu, Ozeki Naoki, Shiba Daisuke, Noda Kousuke, Ishida Susumu, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 7 ) 4143 - 4150 2011.06

ISSN 0146-0404

Purpose. To investigate the influence of deficiency in superoxide dismutase (SOD) 1, a major antioxidative enzyme, on retinal ganglion cells (RGCs). Methods. In the SOD1 total knockout (SOD1-deficient) mice, the level of superoxide anion was measured using dihydroethidium. The number of RGCs was counted in both the retinal sections and the flat-mount retinas after retrograde labeling. Thickness of nerve fiber layer (NFL) was measured in the sections, and the amount of neurofilament protein was measured by immunoblot analysis. Pattern electroretinogram (ERG), which reflects the function of retinal ganglion cells, dark-adapted ERG, and cone ERG were performed. The intraocular pressure (IOP) was measured with an induction-impact tonometer. The levels of SOD-1 and -2 were measured by ELISA, in the serum of 47 newly diagnosed consecutive normal tension glaucoma (NTG) patients and 44 consecutive control subjects. Results. The level of superoxide anion in the RGC layer was significantly higher in 24-week-old SOD1-deficient mice than in wild-type mice. The RGC number was significantly reduced in 24-week-old SOD1-deficient mice, although they were not in 8-week-old mice. The NFL thickness and neurofilament protein were reduced in 24-week-old SOD1-deficient mice. The amplitude of pattern ERG was significantly reduced, although dark-adapted and cone ERGs showed no impairment, in 24-week-old SOD1-deficient mice. The IOP level was not changed in the SOD1-deficient mice. The serum level of SOD1, but not SOD2, was significantly lower in the NTG patients than in the healthy controls. Conclusions. SOD1 deficiency causes RGC vulnerability, which may be involved in the underlying condition of NTG. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Age-related changes in functional visual acuity in healthy individuals

Kaido Minako, Toda Ikuko, Ishida Reiko, Konagai Mototaka, Dogru Murat, Tsubota Kazuo

Japanese Journal of Ophthalmology 55 ( 3 ) 183 - 189 2011.05

ISSN 0021-5155

Objectives: To investigate the age-related alterations in functional visual acuity (FVA) in healthy individuals. Design: A prospective non-comparative multi-center study. Setting and participants: Fifty-four eyes of 54 normal subjects with best corrected visual acuities better than 20/20 seen at the outpatient clinics of the Department of Ophthalmology at Keio University, Minamiaoyama Eye Clinic and Ishida Eye Clinic were studied. Measurements: FVA measurements, Schirmer test, BUT, tear clearance rate and fluorescein vital staining scores were obtained. Results: Schirmer test scores, BUT values and fluorescein vital staining scores did not show significant changes (p > 0.05), but there were significant differences in tear clearance rates among the age groups (p < 0.05). Tear function index, which is a value obtained from multiplying a Schirmer test score by the tear clearance rate, also showed significant differences in the age groups (p < 0.05). The mean logMAR FVA score decreased significantly in the older groups (p < 0.05), while the mean conventional VA score did not show significant differences among the age groups. Moreover, there were significantly more eyes with lower minimum VA scores in the older age groups (p < 0.05). LogMAR minimum VA appeared to be significantly related to the tear clearance rates in subjects 60 years and older (p < 0.05). Conclusions: FVA testing seems to be a useful method of assessing the age-related decline in dynamic visual and tear functions. © 2011 Japanese Ophthalmological Society.

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Clusterin promotes corneal epithelial cell growth through upregulation of hepatocyte growth factor by mesenchymal cells in vitro

Okada Naoko, Kawakita Tetsuya, Mishima Kenji, Saito Ichiro, Miyashita Hideyuki, Yoshida Satoru, Shimmura Shigeto, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 6 ) 2905 - 2910 2011.05

ISSN 0146-0404

PURPOSE. Although the cornea expresses high levels of clusterin (CLU), the role of CLU in the cornea is poorly understood. This study was performed to investigate the possible role of CLU in corneal epithelial homeostasis. METHODS. CLU was overexpressed in 3T3 cells by transfection of a vector encoding full-length CLU (Clu-3T3). Colony-forming efficacy (CFE) was compared in mouse corneal cell line (TKE2) and human primary corneal/limbal epithelial cells that were cocultured with Clu-3T3 and mock-3T3. To determine whether feeder cells have a contact effect, cocultures without feederepithelium contact were also performed. Neutralizing antibody against CLU was used to assess the effects of secretory CLU in TKE2 cells cocultured with Clu-3T3 cells. The expression of growth factors associated with limbal stem/progenitor cell maintenance and growth were analyzed by RT-PCR and Western blot analysis. RESULTS. TKE2 cells cocultured with Clu-3T3 feeders showed higher CFE and were larger in colony size than TKE2 cells cocultured with mock-3T3 feeders. Increased CFE of TKE2 was observed without direct contact with Clu-3T3 cells, which was significantly blocked by treatment with CLU neutralizing antibody. Clu-3T3 cells expressed higher levels of HGF than mock- 3T3 cells, which were significantly suppressed with anti-HGF neutralizing antibodies. Collectively, the promotion of colonyforming and cell proliferation by Clu-3T3 cells was partially mediated by the induction of HGF. CONCLUSIONS. Clusterin indirectly enhances the CFE of corneal/ limbal epithelial cells by inducing the production of HGF by feeder cells, suggesting a role in epithelial-mesenchymal interaction. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Repair of blowout orbital floor fracture by periosteal suturing

Noda Mika, Noda Kousuke, Ideta Shinji, Nakamura Yasuhisa, Ishida Susumu, Inoue Makoto, Tsubota Kazuo

Clinical and Experimental Ophthalmology 39 ( 4 ) 364 - 369 2011.05

ISSN 1442-6404

The study objective was to evaluate the outcome of a periosteal suturing technique in eyes with orbital blowout fractures. Fifteen orbital floor fractures were classified into: type 1, linear fracture; type 2, middle fracture; and type 3, posterior fracture extending two-thirds into the orbit. The feasibility of periosteal suturing without silicone plates or one of minimal size was determined. The improvement of ocular movements and surgical complications were evaluated. A complete suture of the torn periosteum without implanting a silicone plate was achieved in 2/2 (100%) type 1 cases, 5/7 (71%) type 2 cases and 0/6 (0%) type 3 cases. A partial fixation was achieved in 3/6 (50%) type 3 cases with an implantation of a silicone plate of approximate one-third of the usual size in one case. Conventional surgery with silicone plates after failed periosteal suture was required in 2/7 (29%) type 2 cases and 3/6 (50%) type 3 cases. Eye movements were improved postoperatively in all eyes, and a complete range of eye movements was achieved in 6/7 (86%) cases with complete periosteal closure, 1/3 (33%) of cases with partial closure and 2/5 (40%) cases without closure. In conclusion, periosteal suturing can minimize the need for silicone plates especially for anterior or middle orbital floor factures. © 2010 The Authors. Clinical and Experimental Ophthalmology © 2010 Royal Australian and New Zealand College of Ophthalmologists.

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Infectious keratitis outbreak after laser in situ keratomileusis at a single laser center in Japan

Yamaguchi Takefumi, Bissen-Miyajima Hiroko, Hori-Komai Yoshiko, Matsumoto Yukihiro, Ebihara Nobuyuki, Takahashi Hiroshi, Tsubota Kazuo, Shimazaki Jun

Journal of Cataract and Refractive Surgery 37 ( 5 ) 894 - 900 2011.05

ISSN 0886-3350

Purpose: To evaluate an outbreak of infectious keratitis after laser in situ keratomileusis (LASIK) at a single laser center in Japan. Setting: Tokyo Dental College, Chiba, Japan. Design: Case series. Methods: Relevant demographic and clinical data were obtained from case records using a standardized multicenter questionnaire at 12 major hospitals. The clinical manifestations, management, and outcomes were analyzed. Results: Thirty-nine eyes of 30 patients developed infectious keratitis after LASIK at the specified clinic. Cases of infection continued to appear over a 5-month period. The most common interval between LASIK and the onset of infection was within 2 weeks (36 eyes, 92.3%). Slitlamp manifestation included granular opacity beneath flap (71.8%), multiple infiltration (66.7%), and epithelial defect (30.8%). Mycobacterium was identified as the causative organism in 9 eyes (23.1%). In most cases, topical amikacin, arbekacin, and erythromycin in addition to fourth-generation fluoroquinolones were effective. Flap amputation was necessary in 10 eyes (25.6%) of 10 patients. Decimal visual acuity at initial presentation was worse than 0.10 in 14 eyes (35.9%), 0.15 to 0.50 in 8 eyes (20.5%), and 0.60 to 0.90 in 7 eyes (17.9%) and was better than 1.00 in 10 eyes (25.6%). Final decimal visual acuity was worse than 0.10 in 2 eyes (5.1%), 0.15 to 0.50 in 5 eyes (12.8%), and 0.60 to 0.90 in 11 eyes (28.2%) and was better than 1.00 in 21 eyes (53.8%). Conclusions: Mycobacterium was identified as the causative organism of this outbreak. This study shows the possibility of an epidemic of post-LASIK infectious keratitis. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned. © 2011 ASCRS and ESCRS.

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The efficacy, sensitivity, and specificity of strip meniscometry in conjunction with tear function tests in the assessment of tear meniscus

Ibrahim Osama M A, Dogru Murat, Ward Samantha K., Matsumoto Yukihiro, Wakamatsu Tais Hitomi, Ishida Katsushi, Tsuyama Atsushi, Kojima Takashi, Shimazaki Jun, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 5 ) 2194 - 2198 2011.04

ISSN 0146-0404

PURPOSE. To evaluate the specificity and sensitivity of strip meniscometry (SM) testing in conjunction with tear function tests in the diagnosis of dry eye (DE) disease and to investigate the effect of SM on reflex tearing. METHODS. One hundred seven left eyes of 107 patients with definite DE disease according to the Japanese DE diagnostic criteria and 68 left eyes of 68 age- and sex-matched control subjects were studied. Tear meniscus height (TMH) measurements, fluorescein tear film break-up time (FTBUT), fluorescein (F) and rose bengal (RB) staining, and Schirmer's test-1 (ST) were also performed. The assessment of reflex tearing before and after SM application was assessed with a graticule scale at the slit lamp and by optical coherence tomography. The sensitivity and specificity of SM alone and in combination with tear function tests were also sought. RESULTS. The SM scores, TMH measurements, FTBUTs, and STs were significantly lower in dry eye patients than in the controls (P < 0.001). The RB and F staining scores were significantly higher in the dry eye group than in the control group (P < 0.001). The meniscometry strips did not induce significant changes in relation to reflex tearing. SM had an acceptable sensitivity and specificity. CONCLUSIONS. SM is a swift, noninvasive, promising method of assessing tear meniscus volume. The combined SM and FTBUT examination appears to be a sensitive approach to the assessment of dry eye disease. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Kato Naoko, Tsubota Kazuo

American Journal of Ophthalmology 151 ( 4 ) 735 - 736 2011.04

ISSN 0002-9394

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The sparkle of the eye

Goto Eiki, Dogru Murat, Sato Enrique Adan, Matsumoto Yukihiro, Takano Yoji, Tsubota Kazuo

American Journal of Ophthalmology 151 ( 4 ) 691 - 696 2011.04

ISSN 0002-9394

Purpose: To measure the sparkle of the human eye evaluated by the intensity of corneal light reflection in normal subjects and dry eye patients to investigate whether ocular surface wetness has an impact on the sparkle of the eye. Design Prospective case-control study. Methods We examined a consecutive series of eight dry eye patients with Sjgren syndrome (SS, 15 eyes), as well as eight normal subjects (16 eyes). The specular corneal surface light reflection was quantitatively measured with an ophthalmic slit-lamp microscope and image capturing system under fixed conditions of light source, incident angle, and detector sensitivity. The intensity of images from subjects' corneal light reflection was quantified with image analysis software along with the measurement of grade of self-reported brilliancy of the eye, corneal fluorescein staining score, tear film break-up time, and Schirmer test value. The intensity of corneal light reflection was also compared before and after dry eye treatment. Results The mean intensity of corneal light reflection was significantly lower in dry eye patients (125.0 ± 40.1) than normal subjects (167.6 ± 36.6, P = .004). Grade of self-reported brilliancy of the eye, corneal fluorescein staining scores, tear film break-up time, and Schirmer test values showed good correlation to the intensity of corneal light reflection. After punctal plug treatment, the intensity of corneal light reflection significantly increased from 125.0 ± 40.1 to 167.2 ± 45.0 (P < .0001). Conclusions The intensity of corneal light reflection representing the sparkle of the eye was significantly more intense in normal subjects compared to dry eye patients, and was increased after punctal plug treatment. The intensity of corneal light reflection appeared to correlate well with tear film stability, volume, and ocular surface desiccation. We showed that tears contributed not only to ocular surface wetness but also to the extent of the light reflection from the eye. © 2011 Elsevier Inc. All rights reserved.

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The effects of 2 week senofilcon-A silicone hydrogel contact lens daily wear on tear functions and ocular surface health status

Dogru Murat, Ward Samantha K., Wakamatsu Tais, Ibrahim Osama, Schnider Cristina, Kojima Takashi, Matsumoto Yukihiro, Ogawa Junko, Shimazaki Jun, Tsubota Kazuo

Contact Lens and Anterior Eye 34 ( 2 ) 77 - 82 2011.04

ISSN 1367-0484

Purpose: To prospectively investigate the effects of 2 week senofilcon A contact lens (CL) daily wear on the functional visual acuity (VA), ocular surface and tear film. Methods: Seventeen right eyes of 17 senofilcon A CL wearers without any ocular or systemic diseases were examined before and 2 weeks after lens wear. Visual acuity measurements, tear evaporation rate, ELISA for tear cytokines, strip meniscometry, tear lipid layer interferometry, tear film break-up time (BUT), in vivo confocal microscopy, corneal sensitivity, ocular surface vital staining, Schirmer I test and brush cytology for MUC5AC mRNA expression were performed before and after CL wear. Results: The best corrected Landolt VA, functional VA parameters, the mean lipid layer interferometry grades, tear evaporation rates, Schirmer test values, vital staining scores and in vivo confocal microscopy parameters did not show any significant differences after 2 weeks of CL wear. The tear film BUT showed a significant decrease together with a significant down regulation of MUC5 AC mRNA expression after CL wear. A statistically significant elevation in the mean tear interleukin (IL)-6 concentration was also observed after 2 weeks of CL wear. Conclusions: Two week senofilcon A daily CL wear seems to be associated with tear instability, a decrease in MUC5AC expression, and elevation of IL-6 in tears without significant alterations in epithelial damage scores or in the morphology or density of in vivo keratoconjunctival cells and nerves. Alterations associated with long term wear and patients with dry eye disease need to be studied in future trials. © 2010 British Contact Lens Association.

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Surgical punctal occlusion with a high heat-energy releasing cautery device for severe dry eye with recurrent punctal plug extrusion

Ohba Emii, Dogru Murat, Hosaka Eri, Yamazaki Asako, Asaga Rie, Tatematsu Yukako, Ogawa Yoko, Tsubota Kazuo, Goto Eiki

American Journal of Ophthalmology 151 ( 3 ) 2011.03

ISSN 0002-9394

• Purpose: To report the rate of recanalization and the efficacy of punctal occlusion surgery with a high heat-energy-releasing cautery device in patients with severe dry eye disease and recurrent punctal plug extrusion. • Design: Prospective, interventional case series. • Methods: Seventy puncta from 44 eyes of 28 dry eye patients underwent punctal occlusion with thermal cautery. All patients had a history of recurrent punctal plug extrusion. A high heat-energy-releasing thermal cautery device (Optemp II V; Alcon Japan) was used for punctal occlusion surgery. Symptom scores, best-corrected visual acuity, fluorescein staining score, rose bengal staining score, tear film break-up time, and Schirmer test values were compared before and 3 months after the surgery. Rate of punctal recanalization also was examined. • Results: Three months after surgical cauterization, symptom score decreased from 3.9 ± 0.23 to 0.56 ± 0.84 (P < .0001). Logarithm of the minimal angle of resolution best-corrected visual acuity improved from 0.11 ± 0.30 to 0.013 ± 0.22 (P = .003). Fluorescein staining score, rose bengal staining score, tear film break-up time, and the Schirmer test value also improved significantly after the surgery. Only 1 of 70 puncta recanalized after thermal cauterization (1.4%). • Conclusions: Punctal occlusion with the high heat-energy-releasing cautery device not only was associated with a low recanalization rate, but also with improvements in ocular surface wetness and better visual acuity. © 2011 Elsevier Inc.

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Clinical findings in a choroideremia patient who underwent vitrectomy for retinal detachment associated with macular hole

Shinoda Hajime, Koto Takashi, Fujiki Keiko, Murakami Akira, Tsubota Kazuo, Ozawa Yoko

Japanese Journal of Ophthalmology 55 ( 2 ) 169 - 171 2011.03

ISSN 0021-5155

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The international workshop on meibomian gland dysfunction

Nichols Kelly K., Foulks Gary N., Bron Anthony J., Glasgow Ben J., Dogru Murat, Tsubota Kazuo, Lemp Michael A., Sullivan David A.

Investigative Ophthalmology and Visual Science 52 ( 4 ) 1922 - 1929 2011.03

ISSN 0146-0404

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The international workshop on meibomian gland dysfunction

Geerling Gerd, Tauber Joseph, Baudouin Christophe, Goto Eiki, Matsumoto Yukihiro, O'Brien Terrence, Rolando Maurizio, Tsubota Kazuo, Nichols Kelly K.

Investigative Ophthalmology and Visual Science 52 ( 4 ) 2050 - 2064 2011.03

ISSN 0146-0404

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Phakic intraocular lens for keratoconus.

Kato Naoko, Toda Ikuko, Hori-Komai Yoshiko, Sakai Chikako, Arai Hiroyuki, Tsubota Kazuo

Ophthalmology 118 ( 3 ) 2011.03

ISSN 0161-6420

Comparison of corneal thickness and haze in DSAEK and penetrating keratoplasty

Uchino Yuichi, Shimmura Shigeto, Yamaguchi Takefumi, Kawakita Tetsuya, Matsumoto Yukihiro, Negishi Kazuno, Tsubota Kazuo

Cornea 30 ( 3 ) 287 - 290 2011.03

ISSN 0277-3740

Purpose: To compare corneal thickness and haze in Descemet stripping automated endothelial keratoplasty (DSAEK) and penetrating keratoplasty (PK). Methods: This study included 30 consecutive patients undergoing DSAEK (18 patients) and PK (12 patients). Central corneal thickness and corneal haze were analyzed longitudinally by Pentacam (Oculus, Wetzlar, Germany) at 2 weeks, 1 month, and 3 months after grafting. Results: The mean age of patients in the DSAEK group was 71.42 ± 9.91 years (range, 55-85 years) and in the PK group was 60.58 ± 16.58 years (range, 31-81 years). Two weeks, 1 month, and 3 months after surgery, the cornea was significantly thicker in the DSAEK group compared with the PK group (P < 0.01). Two weeks, 1 month, and 3 months after the operation, the cornea had significantly more haze in the DSAEK group compared with the PK group (P < 0.05). Haze in the DSAEK group was mainly observed in the subepithelial stroma. Conclusions: DSAEK grafts were significantly thicker with increased haze for up to 3 months after surgery. Scheimpflug system is available for quantifying not only corneal thickness but also corneal haze after various keratoplasty procedures. Copyright © 2011 by Lippincott Williams & Wilkins.

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[Risk factors of severe peripheral corneal ulcers].

Suzuki Azu, Matsumoto Yukihiro, Uchino Yuichi, Kawakita Tetsuya, Shimmura Shigeto, Tsubota Kazuo

Nippon Ganka Gakkai zasshi 115 ( 2 ) 116 - 121 2011.02

ISSN 0029-0203

To evaluate the risk factors of severe peripheral corneal ulcers. Twenty three eyes of 15 patients (9 eyes of 7 men and 14 eyes of 8 women) with severe peripheral corneal ulcers, who were treated during the past 10 years at Keio University Hospital, were reviewed about the original disease, status of ulcer, treatment, and visual prognosis. Fifteen eyes of 10 patients with Mooren's ulcer, 4 eyes of 3 patients with rheumatoid arthritis, and 4 eyes of 2 patients with Terrien corneal degeneration were included. Mooren's ulcer tends to advance widely. Systemic administration of immunosuppressive and corticosteroidal agents was required in addition to surgical treatment. Visual prognosis was poor in the Mooren's ulcer and perforated cases. Mooren's ulcer and perforated cases are risk factors in peripheral corneal ulcer.

Pharmacotherapy of dry eye

Dogru Murat, Tsubota Kazuo

Expert Opinion on Pharmacotherapy 12 ( 3 ) 325 - 334 2011.02

ISSN 1465-6566

Introduction: Based on data from the largest studies of dry eye to date-the Women's Health Study (WHS) and the Physicians' Health Study (PHS)-and other studies, it has been estimated that about 3.23 million women and 1.68 million men, for a total of 4.91 million Americans aged ≥ 50 years, have dry eye. Tens of millions more have less severe symptoms and probably a more episodic manifestation of the disease that is notable only during contact with some adverse contributing factor(s), such as low humidity or contact lens wear. Dry eye disease is a common yet frequently under-recognized public health problem whose etiology and management challenge clinicians and researchers involved in this field. Areas covered: Advances in the understanding of the disease have been made over the past 10 years in areas of epidemiology, pathogenesis, clinical manifestation, and possible therapy. Historical aspects and recent information in relation to the use of artificial tear substitutes and anti-inflammatory agents in dry eye disease, including topical cyclosporin and corticosteroids, autologous serum, tetracyclines and systemic immunosuppressants, are covered in this review. The reader will gain insight into the recent views on the pharmacological menu of treatments for dry eyes following the recommendations of the 2007 International Dry Eye Workshop. Expert opinion: Dry eye is a visually disabling disease, the treatment of which needs tailoring according to the type and severity of dry eye disease. © 2011 Informa UK, Ltd.

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Functional visual acuity after neodymium

Wakamatsu Tais Hitomi, Yamaguchi Takefumi, Negishi Kazuno, Kaido Minako, Matsumoto Yukihiro, Ishida Reiko, Kojima Takashi, Ibrahim Osama Mohamed Aly, Saiki Megumi, Dogru Murat, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 37 ( 2 ) 258 - 264 2011.02

ISSN 0886-3350

Purpose: To evaluate the changes in functional visual acuity before and after neodymium:YAG (Nd:YAG) laser capsulotomy. Setting: Keio University Hospital, Tokyo, Japan. Design: Case series. Methods: Eyes that had previous cataract surgery with a clinical diagnosis of central posterior capsule opacity requiring Nd:YAG laser capsulotomy were evaluated. All patients had refractive error and corrected distance visual acuity (CDVA) measurements; slitlamp microscopy examinations, including posterior capsule opacification evaluation; high-contrast visual acuity measured at 5 m using a Landolt chart; 10% low-contrast visual acuity (LCVA); functional visual acuity; wavefront examination; and a Schirmer test without anesthesia. Results: The study enrolled 9 patients (4 women, 5 men; 10 eyes) with a mean age of 57.3 years ± 7.0 (SD) (range 47 to 68 years). Although there was no significant difference in CDVA before and after Nd:YAG capsulotomy, there was a significant improvement in the mean functional visual acuity, from 0.18 ± 0.11 logMAR (range -0.03 to 0.31 logMAR) to 0.08 ± 0.07 logMAR (range -0.04 to 0.18 logMAR) (P<.05). The LCVA improved significantly after Nd:YAG laser capsulotomy (P=.003). Higher-order aberrations and the root mean square of the 4th-order coefficients decreased significantly after capsulotomy, and these aberrations were significantly correlated with functional visual acuity and LCVA. Conclusion: Results indicate that measurement of functional visual acuity is useful in assessing the quality of vision in patients who have Nd:YAG laser capsulotomy after cataract surgery. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned. © 2011 ASCRS and ESCRS.

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Hydrogen and N-acetyl-L-cysteine rescue oxidative stress-induced angiogenesis in a mouse corneal alkali-burn model

Kubota Miyuki, Shimmura Shigeto, Kubota Shunsuke, Miyashita Hideyuki, Kato Naoko, Noda Kousuke, Ozawa Yoko, Usui Tomohiko, Ishida Susumu, Umezawa Kazuo, Kurihara Toshihide, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 1 ) 427 - 433 2011.01

ISSN 0146-0404

PURPOSE. To investigate the role of reactive oxygen species (ROS) as the prime initiators of the angiogenic response after alkali injury of the cornea and observe the effects of antioxidants in preventing angiogenesis. METHODS. The corneal epithelia of SOD-1-deficient mice or wild-type (WT) mice were removed after application of 0.15 N NaOH to establish the animal model of alkali burn. ROS production was semiquantitatively measured by dihydroethidium (DHE) fluorescence. Angiogenesis was visualized by CD31 immunohistochemistry. The effects of the specific NF-κB inhibitor DHMEQ, the antioxidant N-acetyl-L-cysteine (NAC), and hydrogen (H₂) solution were observed. RESULTS. ROS production in the cornea was enhanced immediately after alkali injury, as shown by increased DHE fluorescence (P< 0.01). NF-κB activation and the upregulation of vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1 (MCP-1) were significantly enhanced (P< 0.01), leading to a significantly larger area of angiogenesis. Angiogenesis in SOD-1^-/- mice corneas were significantly higher in WT mice (P<0.01), confirming the role of ROS. Pretreatment with the specific NF-κB inhibitor DHMEQ or the antioxidant NAC significantly reduced corneal angiogenesis by downregulating the NF-κB pathway (P<0.01) in both WT and SOD-1^-/- mice. Furthermore, we showed that irrigation of the cornea with hydrogen (H₂) solution significantly reduced angiogenesis after alkali-burn injury (P<0.01). CONCLUSIONS. Immediate antioxidant therapy with H₂-enriched irrigation solution is a new potent treatment of angiogenesis in cornea to prevent blindness caused by alkali burn. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Functional visual acuity measurement in cataract and intraocular lens implantation

Yamaguchi Takefumi, Negishi Kazuno, Tsubota Kazuo

Current Opinion in Ophthalmology 22 ( 1 ) 31 - 36 2011.01

ISSN 1040-8738

Purpose of review: Recently, functional visual acuity (FVA) measurement has been reported to be an important method of determining 'masked impairment of visual function' and assessing visual acuity in detail. This review discusses a new continuous FVA measurement system for the assessment of visual performance and introduces the application of FVA measurement in cataract and intraocular lens implantation. Recent findings: The FVA was first developed to detect impaired visual function for daily activities in dry eye syndrome. The usefulness of this new methodology has been described in patients with tear instability, undergoing laser-assisted in-situ keratomileusis surgery, with Stevens-Johnson syndrome, Sjögren syndrome and mild cataract opacities. The microstructure of the lenticular opacities is highly variable (nucleus sclerosis and/or posterior capsular cataract and/or cortical opacity) causing a combination of different effects on the incident light rays that can be refracted, reflected, absorbed or scattered. In some cases, patients have a good visual acuity but may complain of disturbances in vision quality. We reported the improvement of FVA after cataract surgery in patients with mild cataracts and visual symptomatology despite a good preoperative and postoperative conventional visual acuity. To detect visual impairment due to cataract, measurement of FVA was useful in the evaluation of vision quality and changes in kinetic vision after phacoemulsification surgery. Summary: FVA measurement is an effective and noninvasive test that reflects the complaints of blurring/glare and postoperative satisfaction in patients with mild cataract. © 2010 Wolters Kluwer Health.

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In vivo evaluation of ocular demodicosis using laser scanning confocal microscopy

Kojima Takashi, Ishida Reiko, Sato Enrique Adan, Kawakita Tetsuya, Ibrahim Osama M A, Matsumoto Yukihiro, Kaido Minako, Dogru Murat, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 52 ( 1 ) 565 - 569 2011.01

ISSN 0146-0404

PURPOSE. To investigate the applicability of in vivo laser scanning confocal microscopy in the diagnosis and follow-up of ocular demodicosis infestation in a prospective controlled study. METHODS. Fifteen right eyes of 15 patients with blepharitis associated with cylindrical dandruff (10 males, 5 females; mean age: 62.9 ± 9 years) and eight right eyes of eight age- and sex-matched control subjects underwent HRTII/RCM, evaluation of ocular symptom scores, tear function tests including vital stainings, Schirmer test, and tear clearance test, and evaluation of mite numbers in the eyelids. RESULTS. In vivo confocal microscopy effectively disclosed the mites in the terminal bulbs of the eyelashes, which were not observed after treatment. Eyelids with demodicosis infestation showed marked inflammatory infiltrates around the meibomian glands and conjunctiva, which cleared with tea tree oil treatment. CONCLUSIONS. Laser scanning confocal microscopy seems to be an efficient noninvasive tool in the diagnosis and follow-up of ocular demodicosis infestation. © 2011 The Association for Research in Vision and Ophthalmology, Inc.

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Interferometry in the evaluation of precorneal tear film thickness in dry eye

Hosaka Eri, Kawamorita Takushi, Ogasawara Yuko, Nakayama Nanami, Uozato Hiroshi, Shimizu Kimiya, Dogru Murat, Tsubota Kazuo, Goto Eiki

American Journal of Ophthalmology 151 ( 1 ) 2011.01

ISSN 0002-9394

Purpose To compare tear film thickness between normal subjects and aqueous tear deficiency dry eye patients by tear interferometry. Design Prospective case-control study. Methods Central precorneal tear film thickness was measured noninvasively using an interference thin-film thickness measurement device (Quore MSPA1100; Mamiya-OP). Tear film thickness of 14 eyes from 14 normal subjects and of 28 eyes from 28 aqueous tear deficiency dry eye patients were compared along with noninvasively measured tear meniscus height, DR-1 (Kowa) dry eye severity grading, fluorescein and rose bengal staining scores, tear film break-up time, and Schirmer test results. Among dry eye patients, 13 eyes underwent punctal occlusion, and tear film thickness was compared before and after the surgery. Results Tear film was significantly thinner in dry eye patients (2.0 ± 1.5 μm) than normal subjects (6.0 ± 2.4 μm; P < .0001). Tear film thickness showed good correlation with other dry eye examinations. After punctal occlusion, tear film thickness increased significantly from 1.7 ± 1.5 μm to 4.9 ± 2.8 μm (P = .001) with the improvement of tear meniscus height, fluorescein and rose bengal staining scores, tear film break-up time, and Schirmer test values. Conclusions Interferometric tear film thickness measurement revealed impaired precorneal tear film formation in aqueous tear deficiency dry eyes and was useful for showing the reconstruction of tear film after punctal occlusion surgery. Interferometry of precorneal tear film may be helpful for the evaluation of aqueous tear deficiency in conjunction with other dry eye examinations.

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Porous silk fibroin film as a transparent carrier for cultivated corneal epithelial sheets

Higa Kazunari, Takeshima Naomi, Moro Fumika, Kawakita Tetsuya, Kawashima Motoko, Demura Makoto, Shimazaki Jun, Asakura Tetsuo, Tsubota Kazuo, Shimmura Shigeto

Journal of Biomaterials Science, Polymer Edition 22 ( 17 ) 2261 - 2276 2011

ISSN 0920-5063

Biological carriers, such as the amniotic membrane and serum-derived fibrin, are currently used to deliver cultivated corneal epithelial sheets to the ocular surface. Such carriers require being transparent and allowing the diffusion of metabolites in order to maintain a healthy ocular surface. However, safety issues concerning biological agents encouraged the development of safer, biocompatible materials as cell carriers. We examined the application of porous silk fibroin films with high molecular permeability prepared by mixing silk fibroin and poly(ethylene glycol) (PEG), and then removal of PEG from the silk-PEG films. Molecular permeability of porous silk fibroin film is higher than untreated silk fibroin film. Epithelial cells were isolated from rabbit limbal epithelium, and seeded onto silk fibroin coated wells and co-cultured with mitomycin C-treated 3T3 fibroblasts. Stratified epithelial sheets successfully engineered on porous silk fibroin film expressed the cornea-specific cytokeratins K3 and K12, as well as the corneal epithelial marker pax6. Basement membrane components such as type-IV collagen and integrin β1 were expressed in the stratified epithelial sheets. Further more, colony-forming efficiency of dissociated cells was similar to primary corneal epithelial cells showing that progenitor cells were preserved. The biocompatibility of fibroin films was confirmed in rabbit corneas for up to 6 months. Porous silk fibroin film is a highly transparent, biocompatible material that may be useful as a carrier of cultivated epithelial sheets in the regeneration of corneal epithelium. © Koninklijke Brill NV, Leiden, 2011.

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Neural degeneration in the retina of the streptozotocin-induced type 1 diabetes model

Ozawa Yoko, Kurihara Toshihide, Sasaki Mariko, Ban Norimitsu, Yuki Kenya, Kubota Shunsuke, Tsubota Kazuo

Experimental Diabetes Research 2011 2011

ISSN 1687-5214

Diabetic retinopathy, a vision-threatening disease, has been regarded as a vascular disorder. However, impaired oscillatory potentials (OPs) in the electroretinogram (ERG) and visual dysfunction are recorded before severe vascular lesions appear. Here, we review the molecular mechanisms underlying the retinal neural degeneration observed in the streptozotocin-(STZ-) induced type 1 diabetes model. The renin-angiotensin system (RAS) and reactive oxygen species (ROS) both cause OP impairment and reduced levels of synaptophysin, a synaptic vesicle protein for neurotransmitter release, most likely through excessive protein degradation by the ubiquitin-proteasome system. ROS also decrease brain-derived neurotrophic factor (BDNF) and inner retinal neuronal cells. The influence of both RAS and ROS on synaptophysin suggests that RAS-ROS crosstalk occurs in the diabetic retina. Therefore, suppressors of RAS or ROS, such as angiotensin II type 1 receptor blockers or the antioxidant lutein, respectively, are potential candidates for neuroprotective and preventive therapies to improve the visual prognosis. © 2011 Yoko Ozawa et al.

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Transcriptional factors associated with epithelial-mesenchymal transition in choroidal neovascularization

Hirasawa Manabu, Noda Kousuke, Noda Setsuko, Suzuki Misa, Ozawa Yoko, Shinoda Kei, Inoue Makoto, Ogawa Yoko, Tsubota Kazuo, Ishida Susumu

Molecular Vision 17 1222 - 1230 2011

Purpose: To investigate the transcriptional factors associated with epithelial-mesenchymal transition (EMT) in choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) Methods: Paraffin sections of CNV obtained from patients with AMD (n=12) were stained for transcriptional factors related to EMT, i.e., Snail, Slug, SIP1, and Twist. As a control, postmortem sections of ocular normal tissue were used. Furthermore, using a human retinal pigment epithelial (RPE) cell line (ARPE-19), reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence microscopy were performed to explore the cellular localization and expression levels of EMT-associated transcriptional factors upon cytokine stimulation. Results: Of 12 specimens, 11 CNV tissues (91.6%) showed staining for Snail localized in cellular nuclei, particularly in those of RPE cells. Snail was strongly co-localized with α-smooth muscle antigen (SMA) in RPE cells. In contrast, postmortem human retina showed no Snail staining in RPE cells. Other transcriptional factors, Slug, Twist and SIP1 were not detected in CNV or normal human retina. In ARPE-19 cells, RT-PCR and immunofluorescence microscopy showed that Snail mRNA was upregulated by transforming growth factor (TGF)-β and VEGF stimulation. Furthermore, TGF-β induced relocalization of Snail to the nucleus in RPE cells. Conclusions: The current data indicate that Snail is a major transcriptional factor for EMT changes of RPE cells in human CNV. © 2011 Molecular Vision.

Four cases of corneal perforation in patients with chronic graft-versus-host disease

Inagaki Emi, Ogawa Yoko, Matsumoto Yukihiro, Kawakita Tetsuya, Shimmura Shigeto, Tsubota Kazuo

Molecular Vision 17 598 - 606 2011

ISSN 1090-0535

Purpose: To report the clinical features and investigate the underlying pathological processes of spontaneous corneal perforation in patients with ocular chronic graft-versus-host disease (cGVHD). Methods: A full ophthalmological evaluation of corneal perforation in four patients with cGVHD was performed. Three of them underwent deep anterior lamellar keratoplasty and samples from two of three patients were used for histopathological analyses. Results: Three patients were successfully treated by corneal transplantation. One patient was treated with a therapeutic soft contact lens, and the wound healed within 2 days. The common clinical features of these patients were (1) the presence of definite dry eye related to cGVHD in 3 of 4 patients and probable dry eye in one patient, (2) a central or paracentral site of corneal ulceration and perforation, with no sign of infection, and (3) prior use of a topical or systemic corticosteroid, and/or topical non-steroidal anti-inflammatory drugs. Immunohistochemical findings revealed an increased number of cluster of differentiation 68⁺ (CD68⁺) macrophages and matrix metalloproteinase 9 (MMP-9) expression in the tissue surrounding the perforation. Conclusions: Our report extends current information on the clinical features and pathological processes of corneal perforation in cGVHD by showing increased MMP-9 expression and the accumulation of CD68⁺ positive macrophages in the affected areas. © 2011 Molecular Vision.

Comparison of telomere length and association with progenitor cell markers in lacrimal gland between sjögren syndrome and non-sjögren syndrome dry eye patients

Kawashima Motoko, Kawakita Tetsuya, Maida Yoshiko, Kamoi Mizuka, Ogawa Yoko, Shimmura Shigeto, Masutomi Kenkichi, Tsubota Kazuo

Molecular Vision 17 1397 - 1404 2011

ISSN 1090-0535

Purpose: Indicators of aging such as disruption of telomeric function due to shortening may be more frequent in dysfunctional lacrimal gland. The aims of this study were to 1) determine the viability of quantitative fluorescence in situ hybridization of telomeres (telo-FISH) for the assessment of telomere length in lacrimal gland in Sjögren and non- Sjögren syndrome patients; and 2) investigate the relationship between progenitor cell markers and telomere length in both groups. Methods: Quantitative fluorescence in situ hybridization with a peptide nucleic acid probe complementary to the telomere repeat sequence was performed on frozen sections from human lacrimal gland tissues. The mean fluorescence intensity of telomere spots was automatically quantified by image analysis as relative telomere length in lacrimal gland epithelial cells. Immunostaining for p63, nucleostemin, ATP-binding cassette, sub-family G, member 2 (ABCG2), and nestin was also performed. Results: Telomere intensity in the Sjögren syndrome group (6,785.0±455) was significantly lower than that in the non- Sjögren syndrome group (7,494.7±477; p=0.02). Among the samples from the non-Sjögren syndrome group, immunostaining revealed that p63 was expressed in 1-3 acinar cells in each acinar unit and continuously in the basal layer of duct cells. In contrast, in the Sjögren syndrome group, p63 and nucleostemin showed a lower level of expression. ABCG2 was expressed in acinar cells in both sjogren and non-Sjogren syndrome. Conclusions: The results of this study indicate that 1) telo-FISH is a viable method of assessing telomere length in lacrimal gland, and 2) telomere length in Sjögren syndrome is shorter and associated with lower levels of expression of p63 and nucleostemin than in non-Sjögren syndrome. © 2011 Molecular Vision.

[Demodex-related marginal blepharitis in Japan].

Kawakita Tetsuya, Kawashima Motoko, Ibrahim Osama, Murato Dogru, Tsubota Kazuo

Nippon Ganka Gakkai zasshi 114 ( 12 ) 1025 - 1029 2010.12

ISSN 0029-0203

Some marginal blepharitis is related to demodex, but this has not yet been reported in Japan. In this study, patients with severe marginal blepharitis with cylindrical dandruff were studied to examine the number of demodex in their cilia. Ten eyes of 10 patients (7 men and 3 women, mean age: 62.9 +/- 9.0 years) with unilateral marginal blepharitis which had cylindrical dandruff in their cilia were studied. Three cilia which had the most cylindrical dandruff were removed from the eyelids for microscopic examination. After cleaning the eyelid margins for 1 week, the examination was repeated. Scoring of itching and foreign body sensation of the patients was performed both before and after the treatment. Demodex folliculorum was detected in the cilia of 8 out of 10 eyes (80%), and 22 cilia out of 30 with cylindrical dandruff. The average number of demodex/cilia was 1.6 +/- 0.9. In all cases, the number of demodex/cillium decreased significantly with the improvement in symptoms and blepharitis. An Increase in the number of demodex might be the pathogen causing blepharitis with cylindrical dandruff, and cleaning of the eyelid margin is effective as a therapeutic method.

"Proud of our new era in internationalism" 15th annual meeting of the Kyoto cornea club, December 4-5, 2009

Tsubota Kazuo, Inoue Yoshitsugu, Oshika Tetsuro

Cornea 29 ( SUPPL. 1 ) 2010.11

ISSN 0277-3740

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Increased serum total antioxidant status and decreased urinary 8-hydroxy-2′-deoxyguanosine levels in patients with normal-tension glaucoma

Yuki Kenya, Murat Dogru, Kimura Itaru, Tsubota Kazuo

Acta Ophthalmologica 88 ( 7 ) 2010.11

ISSN 1755-375X

Purpose:- To investigate the amount of systemic oxidative stress-related DNA damage and serum total antioxidant status of patients with normal-tension glaucoma compared with healthy controls. Methods:- Forty-three patients with normal-tension glaucoma selected from 60 consecutive newly diagnosed patients with primary open-angle glaucoma were enrolled. Each patient's intraocular pressure (IOP) was measured seven times over a 24-hour period. Those whose highest IOP was over 21mmHg were excluded. Forty control subjects were recruited. The serum total antioxidant status was assessed colorimetrically by its ability to reduce the generation of the radical cation ABTS (2,2-azinobis-3-ethylbenzthiazoline sulphonate). The urinary 8-hydroxy-2′-deoxyguanosine was measured as a marker of oxidative DNA damage and normalized to creatinine. The results were compared between healthy subjects and patients with normal-tension glaucoma. Results:- The control and normal-tension glaucoma groups did now show significant differences for age, gender, serum fructosamine, cholesterol and triglyceride levels; systolic and diastolic blood pressure, body mass index; and prevalence of hypertension. The serum total antioxidant status was significantly higher (p=0.01) and the urinary 8-hydroxy-2′-deoxyguanosine/creatinine level significantly lower (p=0.03) in the patients with normal-tension glaucoma compared with the controls. Even after we adjusted the data for independent variables, the incidence of normal-tension glaucoma was significantly correlated with high serum total antioxidant levels (p=0.03; odds ratio, 1.007; 95% confidence interval, 1.001-1.013) and low urinary 8-hydroxy-2′-deoxyguanosine/creatinine (p=0.02; odds ratio 0.76; 95% confidence interval, 0.61-0.96). Conclusions:- Increased serum total antioxidant and decreased 8-hydroxy-2′- deoxyguanosine may reflect compensatory alterations in response to increased systemic oxidative stress in patients with normal-tension glaucoma. © 2010 Acta Ophthalmol.

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Immune processes and pathogenic fibrosis in ocular chronic graft-versus-host disease and clinical manifestations after allogeneic hematopoietic stem cell transplantation

Ogawa Yoko, Shimmura Shigeto, Dogru Murat, Tsubota Kazuo

Cornea 29 ( SUPPL. 1 ) 2010.11

ISSN 0277-3740

Activation of the immune system and progressive fibrosis are prominent features of chronic graft-versus-host disease (cGVHD) after allogeneic hematopoietic stem cell transplantation. The main histologic findings in affected exocrine glands are marked fibrosis of the interstitium and prominent increase in the number of fibroblasts, accompanied by mild lymphocytic infiltration. Clinically, the severity of dry eye is correlated with the degree of fibrotic changes, rather than with the amount of lymphocytic infiltration, indicating that excessive extracellular matrix accumulation primarily contributes to the exocrine dysfunction. We have found that accumulated fibroblasts in the lacrimal gland of patients with cGVHD have a chimeric status. Thus, fibroblasts originating from circulating donor-derived precursors and recipient-derived fibroblasts by local epithelial mesenchymal transition may participate in the excessive fibrosis in patients with cGVHD by interacting with T cells. In this article, the immune response and pathogenic fibrosis in ocular cGVHD will be discussed. In addition, clinical findings of ocular complications after hematopoietic stem cell transplantation and currently available treatment will also be discussed. © 2010 by Lippincott Williams & Wilkins.

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The anti-oxidative role of ABCG2 in corneal epithelial cells

Kubota Miyuki, Shimmura Shigeto, Miyashita Hideyuki, Kawashima Motoko, Kawakita Tetsuya, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 51 ( 11 ) 5617 - 5622 2010.11

ISSN 0146-0404

PURPOSE. ABCG2 is a putative marker of progenitor cells, including the corneal epithelium. The authors investigated whether ABCG2 functions in the homeostasis of corneal epithelial cells using abcg2 knockout (KO) mice and corneal epithelial cell lines. METHODS. abcg2 KO mice and a spontaneously immortalized murine corneal epithelial cell line (TKE2) were used for experiments. Flow cytometry was used to determine the presence of side population (SP) cells based on the ability of ABCG2 to efflux Hoechst 33342 dye. Expression of ABCG2 was also examined by RT-PCR. Cytotoxicity assay (IC ₅₀) and propidium iodide staining were performed in semiconfluent cells treated with hypoxia (1% O ₂) or with the pro-oxidant mitoxantrone. RESULTS. abcg2 KO mice had a normal corneal epithelial phenotype; however, cultured abcg2 KO epithelial cells were prone to oxidative damage by mitoxantrone. TKE2 cells were resistant to mitoxantrone at low doses, but higher concentrations were toxic in a dose-dependent manner. Coculture with the ABCG2 inhibitors reserpine and Ko143 inhibited resistance to mitoxantrone, with a statistically higher cell death ratio. abcg2 KO cells were also significantly more sensitive to hypoxia than were wild-type control cells. CONCLUSIONS. ABCG2 may protect corneal epithelial progenitor SP cells against oxidative stress induced by toxins and hypoxia. © Association for Research in Vision and Ophthalmology.

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Simultaneous deep anterior lamellar keratoplasty and limbal allograft in bilateral limbal stem cell deficiency

Omoto Masahiro, Shimmura Shigeto, Hatou Shin, Ichihashi Yoshiyuki, Kawakita Tetsuya, Tsubota Kazuo

Japanese Journal of Ophthalmology 54 ( 6 ) 537 - 543 2010.11

ISSN 0021-5155

Purpose: To report the efficacy of simultaneous keratolimbal allograft (KLAL) surgery and deep anterior lamellar keratoplasty (DALK) for limbal stem cell deficiency (LSCD). Methods: We conducted a retrospective, interventional case series of six consecutive eyes of five patients with LSCD and stromal opacity due to gelatinous drop-like dystrophy (two eyes), Stevens-Johnson syndrome (SJS, two eyes), or aniridia (two eyes). Only patients with normal lid anatomy and Schirmer test values greater than 3 mm were enrolled. DALK was performed by viscodissection followed by a thin, 360° KLAL designed by using an artificial anterior chamber. KLAL sutures were removed after 2 weeks. Results: DALK and KLAL were successfully performed in all eyes, which were followed for an average of 17.2 ± 10.8 months. All eyes recovered a smooth corneal epithelium, although one SJS patient developed a persistent epithelial defect (PED) leading to opacification of the central cornea. Visual acuity improved by more than 2 lines in all eyes except that of the SJS patient with PED. No other complications were observed. Conclusion: Simultaneous DALK and thin-section KLAL is an effective treatment for ocular surface disease in patients with residual tear function and normal lid anatomy. © 2010 Japanese Ophthalmological Society (JOS).

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Application of visante optical coherence tomography tear meniscus height measurement in the diagnosis of dry eye disease

Ibrahim Osama M A, Dogru Murat, Takano Yoji, Satake Yoshiyuki, Wakamatsu Tais Hitomi, Fukagawa Kazumi, Tsubota Kazuo, Fujishima Hiroshi

Ophthalmology 117 ( 10 ) 1923 - 1929 2010.10

ISSN 0161-6420

Purpose: To investigate the applicability of tear meniscus height (TMH) measurement using Visante optical coherence tomography (OCT) in the diagnosis of dry eye disease. Design: Prospective, controlled, single-center study. Participants: Twenty-four right eyes of 24 patients (6 males, 18 females; mean age, 63.14±13.4 years) with definite dry eye according to the Japanese dry eye diagnostic criteria and 27 right eyes of 27 control subjects (12 males, 15 females; mean age, 56.04±14.22 years) were recruited. Methods: All subjects underwent slit-lamp TMH measurement, OCT upper and lower TMH measurements, tear film breakup time (BUT) measurements, vital stainings, and Schirmer test. The results were compared between the 2 groups by Mann-Whitney test. Main Outcome Measures: The correlation between the clinical findings of slit-lamp TMH, strip meniscometry examination, tear functions, vital staining scores, and the OCT upper and lower TMH parameters were tested by Spearman's correlation test. Receiver operating characteristic (ROC) curve technique was used to evaluate the sensitivity, specificity and cutoff values of OCT TMH examination in the diagnosis of dry eye. Results: The OCT upper and lower TMH values, slit-lamp TMH, strip meniscometry, tear film BUT, and vital staining scores were significantly lower in the dry eye patients compared with controls (P<0.001). A significant correlation between the OCT upper and lower TMH measurements as well as slit-lamp TMH, strip meniscometry, tear functions, vital staining scores, and the Schirmer test was found. The ROC curve technique analysis of the OCT lower TMH showed that, when the cutoff value was set at <0.30 mm, the sensitivity and specificity of the testing were 67% and 81%, respectively. Conclusions: The Visante OCT is a quick, noninvasive method for assessing the TMH, with acceptable sensitivity, specificity, and repeatability, and may have potential applications for the diagnosis and evaluation of dry eye disease. Financial Disclosure(s): The authors have no proprietary or commercial interest in any of the materials discussed in this article. © 2010 American Academy of Ophthalmology.

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Comparison of anterior and posterior corneal surface irregularity in descemet stripping automated endothelial keratoplasty and penetrating keratoplasty

Yamaguchi Takefumi, Negishi Kazuno, Yamaguchi Kazuko, Dogru Murat, Uchino Yuichi, Shimmura Shigeto, Tsubota Kazuo

Cornea 29 ( 10 ) 1086 - 1090 2010.10

ISSN 0277-3740

Purpose: To evaluate the irregularity of the anterior and posterior cornea after Descemet stripping automated endothelial keratoplasty (DSAEK) and penetrating keratoplasty (PK). Methods: This clinical study comprised 39 eyes: 13 consecutive eyes after DSAEK, 13 consecutive eyes after PK, and 13 age-matched normal eyes. Corneal elevation data were acquired using a rotating Scheimpflug camera 1 and 3 months after DSAEK and PK. Anterior and posterior corneal elevation data were decomposed into a set of Zernike polynomials up to eighth order. Total higher-order root mean square (RMS) and RMS from third to eighth order were calculated. The astigmatism and irregularity of the anterior and posterior surfaces were compared between DSAEK and PK. Results: The regular astigmatism and tilt components of the anterior surface were significantly lower after DSAEK than after PK at 1 and 3 months (P < 0.001), whereas there was no difference in astigmatism of the posterior surface between the groups (P = 0.07, 0.22). The higher-order RMS and RMSs of third- to eighth-order components of the anterior surface were significantly larger after PK than those after DSAEK at 1 and 3 months (P < 0.01), whereas there were no significant differences between DSAEK and PK in higher-order aberration RMS and RMSs of third- to eighth-order components of the posterior surface. Conclusions: Postoperative corneal irregularity of the anterior surface was greater after PK than after DSAEK, whereas there was no significant difference in posterior surface irregularity. DSAEK is superior to PK in terms of the higher-order irregularity of the anterior surface. Copyright © 2010 by Lippincott Williams & Wilkins.

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Topography-guided conductive keratoplasty

Kato Naoko, Toda Ikuko, Kawakita Tetsuya, Sakai Chikako, Tsubota Kazuo

American Journal of Ophthalmology 150 ( 4 ) 2010.10

ISSN 0002-9394

Purpose: To evaluate the use of topography-guided conductive keratoplasty in eyes with keratoconus. Design: Interventional case series. Methods: We examined 21 eyes in 21 patients with advanced keratoconus. Topography-guided conductive keratoplasty was performed with intraoperative monitoring of corneal astigmatism using a surgical keratometer. Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), corneal topography, manifest refraction, intraocular pressure (IOP), corneal endothelial cell counts, complications, and eventual outcomes were evaluated. Results: UCVA (logarithm of the minimal angle of resolution [logMAR]), which was 1.65 ± 0.49 preoperatively, improved to 1.04 ± 0.64 at 1 week (P < .001) and 1.12 ± 0.61 at 1 month after surgery (P < .001). BSCVA, which was 1.02 ± 0.56 preoperatively, improved to 0.76 ± 0.65 at 1 week (P = .026) and 0.76 ± 0.60 at 1 month after surgery (P = .003). Manifest refraction, which was -15.13 ± 6.66 diopters (D) before surgery, declined to -9.97 ± 6.71 D at 1 month after surgery (P = .002). Although corneal topography reverted to the preoperative pattern and UCVA and BSCVA also regressed toward preoperative values, 12 of 21 eyes were better able to tolerate and conduct normal daily activities using contact lenses. Five subjects have undergone or are considering corneal transplantation after unsatisfactory postoperative results. No serious perioperative complication was observed. Conclusions: Topography-guided conductive keratoplasty may be effective in reshaping corneal configuration in eyes with keratoconus, without serious complications, and possibly contributed to avoiding or delaying corneal transplantation. © 2010 Elsevier Inc. All Rights Reserved.

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Resveratrol prevents light-induced retinal degeneration via suppressing activator protein-1 activation

Kubota Shunsuke, Kurihara Toshihide, Ebinuma Mari, Kubota Miyuki, Yuki Kenya, Sasaki Mariko, Noda Kousuke, Ozawa Yoko, Oike Yuichi, Ishida Susumu, Tsubota Kazuo

American Journal of Pathology 177 ( 4 ) 1725 - 1731 2010.10

ISSN 0002-9440

Light damage to the retina accelerates retinal degeneration in human diseases and rodent models. Recently, the polyphenolic phytoalexin resveratrol has been shown to exert various bioactivities in addition to its classical antioxidant property. In the present study, we investigated the effect of resveratrol on light-induced retinal degeneration together with its underlying molecular mechanisms. BALB/c mice with light exposure (5000-lux white light for 3 hours) were orally pretreated with resveratrol at a dose of 50 mg/kg for 5 days. Retinal damage was evaluated by TdT-mediated dUTP nick-end labeling, outer nuclear layer morphometry, and electroretinography. Administration of resveratrol to mice with light exposure led to a significant suppression of light-induced pathological parameters, including TdT-mediated dUTP nick-end labeling-positive retinal cells, outer nuclear layer thinning, and electroretinography changes. To clarify the underlying molecular mechanisms, the nuclear translocation of activator protein -1 subunit c-fos was evaluated by enzyme-linked immunosorbent assay, and the retinal activity of sirtuin 1 was measured by deacetylase fluorometric assay. Retinal activator protein-1 activation, up-regulated following light exposure, was significantly reduced by application of resveratrol. In parallel, retinal sirtuin 1 activity, reduced in animals with light damage, was significantly augmented by resveratrol treatment. Our data suggest the potential use of resveratrol as a therapeutic agent to prevent retinal degeneration related to light damage. Copyright © American Society for Investigative Pathology.

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Elevated serum immunoglobulin G titers against chlamydia pneumoniae in primary open-angle glaucoma patients without systemic disease

Yuki Kenya, Kimura Itaru, Shiba Daisuke, Imamura Yutaka, Tsubota Kazuo

Journal of Glaucoma 19 ( 8 ) 535 - 539 2010.10

ISSN 1057-0829

Purpose: To determine if Chlamydia pneumoniae infection is associated with primary open-angle glaucoma (POAG). Patients and Methods: Consecutive patients with newly diagnosed primary POAG attending the Glaucoma clinic of Keio University Hospital between June 2007 and January 2008 were considered for inclusion in this prospective case-control study. Forty consecutive POAG patients and 41 normal healthy individuals as a control population met the inclusion criteria. The exclusion criteria for both groups were; taking steroids or immunosuppressive agents, smoking, and history of any acute or chronic systemic disease including stroke, heart attack, diabetes mellitus, hypertension, asthma, and autoimmune diseases. The serum was analyzed for C. pneumoniae and C. trachomatis immunoglobulin G antibody titers by enzyme-linked immunosorbent assay. Seroactivity to each antigen between case and control groups was evaluated by Mann-Whitney U test. Results: The age, male/female ratio, and intraocular pressure of the cases and control groups were not significantly different. Immunoglobulin G titers for C. pneuemoniae was significantly higher in patients with POAG than in controls (P=0.009). The titers to C. trachomatis were not significantly different between the 2 groups (P=0.99). Conclusion: The results suggest that higher C. pneumoniae titers are associated with POAG. If confirmed, this may indicate either a common factor that causes susceptibilities to both glaucoma and C. pneumoniae infection or that C. pneumoniae may be a causal factor for developing POAG. Copyright © 2010 by Lippincott Williams & Wilkins.

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Retinal aging and sirtuins

Ozawa Yoko, Kubota Shunsuke, Narimatsu Toshio, Yuki Kenya, Koto Takashi, Sasaki Mariko, Tsubota Kazuo

Ophthalmic Research 44 ( 3 ) 199 - 203 2010.09

ISSN 0030-3747

The process of aging involves the accumulating changes in the microenvironment that lead to cell senescence or apoptosis, and subsequent tissue or organ dysfunction. Multiple extrinsic and intrinsic events that cause DNA instability are associated with aging. Cells containing unstable DNA are biologically vulnerable, and if the DNA damage is too great for the cell to repair, it becomes senescent or dies by apoptosis. Thus, the cell's capacity to repair its DNA determines the progress of aging, at least in part. Here, we focus on the sirtuins, the mammalian homologs of the yeast life-span-extending molecule, Sir2. Among the sirtuin family proteins in mammals, the one most similar to yeast Sir2 is SIRT1, which is involved in multiple pathways, including the repair of DNA double-strand breaks. Although the role of SIRT1 in mammalian longevity is not clear, it is expressed throughout the retina, where it may suppress aging. In fact, a mutant mouse model of retinal degeneration shows an abnormal subcellular localization of SIRT1 protein and accelerated retinal cell apoptosis. Further analyses are required to elucidate the mechanism of DNA damage and repair, including the contributions of the sirtuins, in the aged or diseased retinas, which will help us understand the mechanisms of retinal aging. © 2010 S. Karger AG, Basel.

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The era of antiaging ophthalmology comes of age

Tsubota Kazuo, Kawashima Motoko, Inaba Takaaki, Dogru Murat, Ogawa Yoko, Nakamura Shigeru, Shinmura Ken, Higuchi Akihiro, Kawakita Tetsuya

Ophthalmic Research 44 ( 3 ) 146 - 154 2010.09

ISSN 0030-3747

Recent advances in the understanding of aging have paved a new way of thinking about intervening with the aging process. There is a global agreement in the scientific community that calorie restriction (CR) can actually extend the life span of various kinds of animals so that this has become a real intervention in aging. In addition to the CR theory, the free radical theory is another important hypothesis, which is believed to be involved in aging. According to this theory, we can manage the aging process by controlling calories or reactive oxygen species. In this paper, these two important aging theories, CR and free radical aging, are reviewed, and it is discussed how to apply these theories to the prevention and treatment of eye diseases. Finally, we share the preliminary results of our animal study on dry eye, and I report my personal experience as a dry eye patient, which has been alleviated by the antiaging approach. © 2010 S. Karger AG, Basel.

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Ophthalmic Research

Tsubota Kazuo

Ophthalmic Research 44 ( 3 ) 145 2010.09

ISSN 0030-3747

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Evaluation of therapeutic effects of astaxanthin on impairments in salivary secretion

Yamada Takashi, Ryo Koufuchi, Tai Yoshinori, Tamaki Yoh, Inoue Hiroko, Mishima Kenji, Tsubota Kazuo, Saito Ichiro

Journal of Clinical Biochemistry and Nutrition 47 ( 2 ) 130 - 137 2010.09

ISSN 0912-0009

The involvement of reactive oxygen species (ROS) in the pathophysiology of Sjögren's syndrome (SS), an autoimmune disorder, and irradiation-induced impairments in salivary secretion has been reported. Meanwhile, the strong antioxidant astaxanthin (Ast) has been suggested to have therapeutic effects on various diseases. In the present study, we examined the ROS scavenging capacity of Ast using a human salivary gland epithelial cell line (HSY) and investigated the effects of Ast on salivary secretion in a mouse model of irradiation-induced salivary gland dysfunction. Furthermore, we performed a clinical study of Ast in six SS patients and six normal individuals, quantifying the volume of saliva secretion and the level of oxidative stress markers in the saliva. Ast partially suppressed hydrogen peroxide-induced ROS in HSY cells. The mouse model demonstrated that the preadministration of Ast resulted in the suppression of irradiation-induced hyposalivation. Furthermore, the administration of Ast appeared to increase salivary output in both the SS and normal groups. The level of oxidative stress marker, hexanoyl-lysine, in the saliva was reduced after Ast intake. These results suggest that Ast might act as an ROS scavenger, providing benefits to SS patients with impaired salivary secretion.

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Dietary restriction

Baur Joseph A., Chen Danica, Chini Eduardo N., Chua Katrin, Cohen Haim Y., De Cabo Rafael, Deng Chuxia, Dimmeler Stefanie, Gius David, Guarente Leonard P., Helfand Stephen L., Imai Shin Ichiro, Itoh Hiroshi, Kadowaki Takashi, Koya Daisuke, Leeuwenburgh Christiaan, Mcburney Michael, Nabeshima Y. O Ichi, Neri Christian, Oberdoerffer Philipp, Pestell Richard G., Rogina Blanka, Sadoshima Junichi, Sartorelli Vittorio, Serrano Manuel, Sinclair David A., Steegborn Clemens, Tatar Marc, Tissenbaum Heidi A., Tong Qiang, Tsubota Kazuo, Vaquero Alejandro, Verdin Eric

Science 329 ( 5995 ) 1012 - 1013 2010.08

ISSN 0036-8075

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In vivo evaluation of superior limbic keratoconjunctivitis using laser scanning confocal microscopy and conjunctival impression cytology

Kojima Takashi, Matsumot Yukihiro, Ibrahim Osama M A, Sato Enrique Adan, Dogru Murat, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 51 ( 8 ) 3986 - 3992 2010.08

ISSN 0146-0404

PURPOSE. To investigate the cytologic findings of superior bulbar conjunctiva in superior limbic keratoconjunctivitis (SLK) using laser scanning confocal microscopy and impression cytology in a prospective controlled study. METHODS. Twenty-one eyes of 11 SLK patients (9 women, 2 men; mean age, 49.3 ± 17.9 years) and 18 eyes of 9 control subjects (6 women, 3 men; mean age, 46.4 ± 8.7 years) underwent tear function tests including vital stainings, Schirmer test, tear clearance test, digital confocal laser scanning microscopy, and conjunctival impression cytology. After confocal microscopy and impression cytology images were obtained, the mean individual epithelial cell area (MIECA), nucleocytoplasmic (N/C) ratio, and inflammatory cell density were analyzed. The correlation between confocal microscopy and impression cytology parameters was investigated. RESULTS. The MIECA of SLK patients and control subjects in confocal microscopy was 786.54 ± 463.88 μm² and 311.50 ± 78.30 μm², respectively. The mean N/C ratio was 0.356 ± 0.090 and 0.490 ± 0.038, respectively. The MIECA and N/C ratio in impression cytology showed significant correlation with the corresponding confocal microscopy parameters (MIECA, P ± 0.0028; N/C, P= 0.0051). The inflammatory cell density in confocal microscopy significantly correlated with superior bulbar conjunctival Rose-Bengal scores (P= 0.0264). CONCLUSIONS. Laser scanning confocal microscopy seems to be an efficient noninvasive tool in the evaluation of phenotypic alterations of the conjunctival epithelium in SLK and may serve as an alternative for impression cytology. N/C ratio and inflammatory cell density appear to be two new promising parameters of in vivo confocal microscopy in the assessment of ocular surface disease in SLK. © Association for Research in Vision and Ophthalmology.

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A new method to prevent loss of mitomycin C soaked sponges under the conjunctiva during trabeculectomy

Yuki Kenya, Shiba Daisuke, Ota Yu, Ozeki Naoki, Murat Dogru, Tsubota Kazuo

British Journal of Ophthalmology 94 ( 8 ) 1111 - 1112 2010.08

ISSN 0007-1161

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Role of insulin in regulation of Na⁺-/K⁺-dependent ATPase activity and pump function in corneal endothelial cells

Hatou Shin, Yamada Masakazu, Akune Yoko, Mochizuki Hiroshi, Shiraishi Atsushi, Joko Takeshi, Nishida Teruo, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 51 ( 8 ) 3935 - 3942 2010.08

ISSN 0146-0404

PURPOSE. The Na⁺-/K⁺-dependent ATPase (Na, K-ATPase) expressed in the basolateral membrane of corneal endothelial cells plays an important role in the pump function of the corneal endothelium. The role of insulin in the regulation of Na,K-ATPase activity and pump function in corneal endothelial cells was investigated. METHODS. Confluent monolayers of mouse corneal endothelial cells were exposed to insulin. ATPase activity was evaluated by spectrophotometric measurement of phosphate released from ATP with the use of ammonium molybdate; Na, K-ATPase activity was defined as the portion of total ATPase activity sensitive to ouabain. Pump function was measured with the use of a Ussing chamber; pump function attributable to Na, K-ATPase activity was defined as the portion of the total short-circuit current sensitive to ouabain. Western blot analysis and immunocytochemistry were performed to measure the expression of the Na,K-ATPase α₁-subunit. RESULTS. Insulin increased the Na, K-ATPase activity and pump function of cultured corneal endothelial cells. These effects were blocked by protein kinase C (PKC) inhibitors and protein phosphatases 1 and 2A inhibitor. Western blot analysis indicated that insulin decreased the ratio of the inactive Na, KATPase α₁-subunit. Immunocytochemistry indicated that insulin increased the cell surface expression of the Na, K-ATPase α₁-subunit. CONCLUSIONS. These results suggest that insulin increases the Na, K-ATPase activity and pump function of cultured corneal endothelial cells. The effect of insulin is mediated by PKC and presumably results in the activation of PP1, 2A, or both, which are essential for activating Na,K-ATPase by α₁-subunit dephosphorylation. © Association for Research in Vision and Ophthalmology.

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Calorie restriction

Kawashima Motoko, Kawakita Tetsuya, Okada Naoko, Ogawa Yoko, Murat Dogru, Nakamura Shigeru, Nakashima Hideo, Shimmura Shigeto, Shinmura Ken, Tsubota Kazuo

Biochemical and Biophysical Research Communications 397 ( 4 ) 724 - 728 2010.07

ISSN 0006-291X

A decrease in lacrimal gland secretory function is closely related to aging and leads to an increased prevalence of dry eye syndrome. Since calorie restriction (CR) is considered to prevent functional decline of various organs due to aging, we hypothesized that CR could prevent age-related lacrimal dysfunction. Six-month-old male Fischer 344 rats were randomly divided into ad libitum (AL) and CR (-35%) groups. After 6. months of CR, tear function was examined under conscious state. After euthanasia, lacrimal glands were subjected to histological examination, tear protein secretion stimulation test with Carbachol, and assessment of oxidative stress with 8-hydroxy-2 deoxyguanosine (8-OHdG) and 4-hydroxynonenal (HNE) antibodies. CR significantly improved tear volume and tended to increase tear protein secretion volume after stimulation with Carbachol compared to AL. The acinar unit density was significantly higher in the CR rats compared to AL rats. Lacrimal glands in the CR rats showed a lesser degree of interstitial fibrosis. CR reduced the concentration of 8-OHdG and the extent of staining with HNE in the lacrimal gland, compared to AL. Furthermore, our electron microscopic observations showed that mitochondrial structure of the lacrimal gland obtained from the middle-aged CR rats was preserved in comparison to the AL rats. Collectively, these results demonstrate for the first time that CR may attenuate oxidative stress related damage in the lacrimal gland with preservation of lacrimal gland functions. Although molecular mechanism(s) by which CR maintains lacrimal gland function remains to be resolved, CR might provide a novel therapeutic strategy for treating dry eye syndrome. © 2010 Elsevier Inc.

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Secondary macular hole formation with presumed evulsion of foveal hard exudates in a patient with diabetic retinopathy

Tomita Yohei, Kousuke Noda, Hajime Shinoda, Yoko Ozawa, Kazuo Tsubota, Susumu Ishida

Japanese Journal of Ophthalmology 54 ( 4 ) 366 - 368 2010.07

ISSN 0021-5155

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Baseline profiles of ocular surface and tear dynamics after allogeneic hematopoietic stem cell transplantation in patients with or without chronic GVHD-related dry eye

Wang Y., Ogawa Y., Dogru M., Tatematsu Y., Uchino M., Kamoi M., Okada N., Okamoto S., Tsubota K.

Bone Marrow Transplantation 45 ( 6 ) 1077 - 1083 2010.06

ISSN 0268-3369

We evaluated ocular surface alterations in allogeneic hematopoietic stem cell transplantation (HSCT) recipients with or without chronic GVHD-related dry eye in a prospective study. Fifty eyes of 25 post-HSCT patients and 28 eyes of 14 age-matched healthy controls were included. Meibomian gland (MG) obstruction, tear evaporation rate, corneal sensitivity (CS), Schirmer test-I, tear break-up time (BUT) and ocular surface vital staining were examined. Conjunctival impression and brush cytology specimens were collected to evaluate the goblet cell density (GCD) and the inflammatory cell numbers. Obvious MG obstruction, decreased CS and enhanced tear evaporation rate were found in post-HSCT patients compared with normal controls. In addition, decreased conjunctival GCD, increased conjunctival squamous metaplasia and inflammatory cells were noted in cGVHD-related dry eyes compared with normal controls and post-HSCT without dry eye subjects. Furthermore, the conjunctival inflammatory cells were significantly higher in severe dry eyes compared with mild dry eyes (P=0.03). We found comprehensive ocular surface alteration in post-HSCT patients, regardless of whether they had cGVHD-related dry eye or not. The results suggest that the extent of inflammatory process seems to have a pivotal role in the outcome of the cGVHD-related dry eye. © 2010 Macmillan Publishers Limited All rights reserved.

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S-1 Induces Meibomian Gland Dysfunction

Matsumoto Yukihiro, Dogru Murat, Sato Enrique Adan, Ibrahim Osama M A, Tatematsu Yukako, Ogawa Yoko, Tsubota Kazuo

Ophthalmology 117 ( 6 ) 2010.06

ISSN 0161-6420

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Efficacy of small-incision intraocular lens exchange of opacified Hydroview implants

Yamaguchi Takefumi, Shimizu Tsutomu, Yamaguchi Kazuko, Dogru Murat, Negishi Kazuno, Tsubota Kazuo

British Journal of Ophthalmology 94 ( 6 ) 808 - 809 2010.06

ISSN 0007-1161

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Implantation of multifocal intraocular lens in two cases after LASIK

Shirahata Reiko, Negishi Kazuno, Torii Hidemasa, Dogru Murat, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 64 ( 6 ) 881 - 884 2010.06

ISSN 0370-5579

Purpose : To report two cases who had received LASIK previously and who underwent cataract surgery with implantation of multifocal intraocular lens (IOL). Cases and Findings : Both were males aged 47 and 55 years respectively. LASIK had been performed 6 years and 2 years before respectively. Both cases were diagnosed with cataract in the right eye. Corrected visual acuity was 1.2 and 0.7 respectively. The affected eye was treated by cataract surgery and implantation of diffractive multifocal IOL. After surgery, near and far visual acuity with the naked eye was 1.2 and 1.0 in one case and 1.0 each in the other. Both patients expressed satisfaction with the outcome. Contrast sensitivity was at the lower limit of normal range in both cases. Conclusion : These two cases illustrate that refractive error and impaired visual function may result following implantation of multifocal IOL after LASIK. We advocate due explanation to obtain informed consent in such cases.

Ocular complications in mulvihill-smith syndrome

Ibrahim O. M A, Takefumi Y., Dogru M., Negishi K., Kosaki K., Tsubota K.

Eye 24 ( 6 ) 1123 - 1124 2010.06

ISSN 0950-222X

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Von Hippel-Lindau protein regulates transition from the fetal to the adult circulatory system in retina

Kurihara Toshihide, Kubota Yoshiaki, Ozawa Yoko, Takubo Keiyo, Noda Kousuke, Simon M. Celeste, Johnson Randall S., Suematsu Makoto, Tsubota Kazuo, Ishida Susumu, Goda Nobuhito, Suda Toshio, Okano Hideyuki

Development 137 ( 9 ) 1563 - 1571 2010.05

ISSN 0950-1991

In early neonates, the fetal circulatory system undergoes dramatic transition to the adult circulatory system. Normally, embryonic connecting vessels, such as the ductus arteriosus and the foramen ovale, close and regress. In the neonatal retina, hyaloid vessels maintaining blood flow in the embryonic retina regress, and retinal vessels take over to form the adult-type circulatory system. This process is regulated by a programmed cell death switch mediated by macrophages via Wnt and angiopoietin 2 pathways. In this study, we seek other mechanisms that regulate this process, and focus on the dramatic change in oxygen environment at the point of birth. The von Hippel-Lindau tumor suppressor protein (pVHL) is a substrate recognition component of an E3-ubiquitin ligase that rapidly destabilizes hypoxia-inducible factor as (HIF-αs) under normoxic, but not hypoxic, conditions. To examine the role of oxygen-sensing mechanisms in retinal circulatory system transition, we generated retina-specific conditional-knockout mice for VHL (Vhl ^α-CreKO mice). These mice exhibit arrested transition from the fetal to the adult circulatory system, persistence of hyaloid vessels and poorly formed retinal vessels. These defects are suppressed by intraocular injection of FLT1-Fc protein [a vascular endothelial growth factor (VEGF) receptor-1 (FLT1)/Fc chimeric protein that can bind VEGF and inhibit its activity], or by inactivating the HIF-1a gene. Our results suggest that not only macrophages but also tissue oxygen-sensing mechanisms regulate the transition from the fetal to the adult circulatory system in the retina. © 2010. Published by The Company of Biologists Ltd.

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Predictability of ocular spherical aberration after cataract surgery determined using preoperative corneal spherical aberration

Negishi Kazuno, Kodama Chiyo, Yamaguchi Takefumi, Torii Hidemasa, Saiki Megumi, Dogru Murat, Ohnuma Kazuhiko, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 36 ( 5 ) 756 - 761 2010.05

ISSN 0886-3350

Purpose: To evaluate the predictability of total postoperative spherical aberration after aspheric intraocular lens (IOL) implantation using preoperative corneal spherical aberration. Setting: Keio University Hospital, Tokyo, Japan. Methods: Corneal and ocular higher-order aberrations (HOAs) were analyzed preoperatively and postoperatively in eyes having cataract extraction with aspheric IOL implantation. The predicted postoperative total spherical aberration Z(4,0) was calculated by adding the preoperative corneal spherical aberration and the labeled spherical aberration of the IOL. The prediction error of the postoperative total spherical aberration was calculated by subtracting the predicted postoperative total spherical aberration from the postoperative total spherical aberration. Surgically induced corneal spherical aberration was calculated by subtracting the postoperative from the preoperative corneal spherical aberration. Results: There were no statistically significant differences between preoperative and postoperative corneal spherical aberration, 3rd-order aberration, or HOAs. The prediction error of the postoperative total spherical aberration was within ±0.05 μm in 9 (24.3%) of the 37 eyes evaluated and within ±0.10 μm in 18 eyes (48.6%). Statistically significantly correlations were found between the surgically induced corneal spherical aberration and prediction error of postoperative total spherical aberration, the preoperative corneal spherical aberration and surgically induced corneal spherical aberration, and the preoperative corneal spherical aberration and prediction error of postoperative total spherical aberration. Conclusions: Predictability of postoperative ocular spherical aberration after cataract surgery with aspheric IOL implantation was insufficient because of the surgically induced corneal spherical aberration. Considering surgically induced corneal spherical aberration could improve the predictability of postoperative ocular spherical aberration. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned. © 2010 ASCRS and ESCRS.

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Comparison of effects of periocular region dry and wet warming on visual acuity and near reflex

Takahashi Yoko, Igaki Michihito, Sakamoto Ichirou, Suzuki Atsushi, Takahashi Genichiro, Dogru Murat, Tsubota Kazuo

Nippon Ganka Gakkai zasshi 114 ( 5 ) 444 - 453 2010.05

ISSN 0029-0203

PURPOSE: To investigate the influence of warming the periocular region for 10 minutes after visual display terminal (VDT) work on the near reflex, and the differences in influence between lid closure, dry and wet warming, and the amounts of water vapor. SUBJECTS AND METHODS: Subjects in the first trial were 10 healthy volunteers (10 eyes). Following VDT, subjects wore either a warming eye steamer (water vapor: 200 mg/10 min) or an eye mask with their eyelids closed for 10 minutes. Subjects in the second trial were 5 healthy volunteers (5 eyes). Following VDT, subjects applied either a dry-warming sheet (water vapor: 0 mg/10 min) or a wet-warming sheet (water vapor: 20, 35, 160 and 200 mg/10 min) to there eyelids and the periocular region for 10 minutes. In both experiments, subjective amplitude of accommodation, pupillary reaction, convergence reaction, near vision, and breakup time were measured before and immediately after VDT work and after 2 experimental settings. RESULTS: The warming eye steamer significantly improved amplitude of accommodation (p<0.05), and near vision (p<0.05). Warming the periocular region with water vapor improved subjective amplitude of accommodation, and near vision, the effect of which depended on the amount of water vapor (p<0.05). CONCLUSIONS: Warming the periocular region with water vapor improved subjective amplitude of accommodation, near vision and the effect depended on the amount of water vapor.

Passive cigarette smoke exposure and soft contact lens wear

Ward Samantha K., Dogru Murat, Wakamatsu Tais, Ibrahim Osama, Matsumoto Yukihiro, Kojima Takashi, Sato Enrique Adan, Ogawa Junko, Schnider Cristina, Negishi Kazuno, Tsubota Kazuo

Optometry and Vision Science 87 ( 5 ) 367 - 372 2010.05

ISSN 1040-5488

PURPOSE.: To prospectively investigate the effects of passive cigarette smoke exposure on the ocular surface and tear film in soft contact lens (SCL) wearers. METHODS.: Twelve right eyes of 12 SCL wearers without any ocular or systemic diseases and 10 right eyes of 10 subjects who never wore CLs were examined before and 2 h after 5 min of passive cigarette smoke exposure in a controlled smoke chamber. Tear evaporation rate measurement, tear film break-up time (TBUT) examination, ocular surface fluorescein, rose bengal stainings, and Schirmer I test were performed at each visit. RESULTS.: The mean tear evaporation rates, TBUTs, and vital staining scores were significantly worse in CL wearers compared with healthy control subjects. TBUTs showed significant worsening after passive smoke exposure in both groups. The mean tear evaporation rate and vital staining scores showed a significant increase with brief passive smoke exposure in subjects not wearing CLs but not in CL wearers. CONCLUSION.: Even brief passive exposure to cigarette smoke is associated with adverse effects on the ocular surface as evidenced by an increase in tear instability and damage to the ocular surface epithelia in SCL wearers and non-CL wearers. Copyright © 2010 American Academy of Optometry.

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Lacrimal Gland in Sjögren's Syndrome

Sato Enrique Adan, Matsumoto Yukihiro, Dogru Murat, Kaido Minako, Wakamatsu Tais, Ibrahim Osama M A, Obata Hiroto, Tsubota Kazuo

Ophthalmology 117 ( 5 ) 2010.05

ISSN 0161-6420

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Neurodegenerative influence of oxidative stress in the retina of a murine model of diabetes

Sasaki M., Ozawa Y., Kurihara T., Kubota S., Yuki K., Noda K., Kobayashi S., Ishida S., Tsubota K.

Diabetologia 53 ( 5 ) 971 - 979 2010.05

ISSN 0012-186X

Aims/hypothesis: Diabetic retinopathy is a progressive neurodegenerative disease, but the underlying mechanism is still obscure. Here, we focused on oxidative stress in the retina, and analysed its influence on retinal neurodegeneration, using an antioxidant, lutein. Methods: C57BL/6 mice with streptozotocin-induced diabetes were constantly fed either a lutein-supplemented diet or a control diet from the onset of diabetes, and their metabolic data were recorded. In 1-month-diabetic mice, reactive oxygen species (ROS) in the retina were measured using dihydroethidium and visual function was evaluated by electroretinograms. Levels of activated extracellular signal-regulated kinase (ERK), synaptophysin and brain-derived neurotrophic factor (BDNF) were also measured by immunoblotting in the retina of 1-month-diabetic mice. In the retinal sections of 4-month-diabetic mice, histological changes, cleaved caspase-3 and TUNEL staining were analysed. Results: Lutein did not affect the metabolic status of the diabetic mice, but it prevented ROS generation in the retina and the visual impairment induced by diabetes. ERK activation, the subsequent synaptophysin reduction, and the BDNF depletion in the diabetic retina were all prevented by lutein. Later, in 4-month-diabetic mice, a decrease in the thickness of the inner plexiform and nuclear layers, and ganglion cell number, together with increase in cleaved caspase-3- and TUNEL-positive cells, were avoided in the retina of lutein-fed mice. Conclusions/interpretation: The results indicated that local oxidative stress that has a neurodegenerative influence in the diabetic retina is prevented by constant intake of a lutein-supplemented diet. The antioxidant, lutein may be a potential therapeutic approach to protect visual function in diabetes. © 2010 Springer-Verlag.

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The role of oxidative stress and inflammation in conjunctivochalasis

Ward Samantha K., Wakamatsu Tais Hitomi, Dogru Murat, Ibrahim Osama M A, Kaido Minako, Ogawa Yoko, Matsumoto Yukihiro, Igarashi Ayako, Ishida Reiko, Shimazaki Jun, Schnider Cristina, Negishi Kazuno, Katakami Chikako, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 51 ( 4 ) 1994 - 2002 2010.04

ISSN 0146-0404

PURPOSE. To investigate the status of oxidative stress and histopathologic alterations in patients with conjunctivochalasis and compare the findings with those in healthy control subjects. METHODS. Eleven patients (n = 20 eyes) with Yokoi grade 3 conjunctivochalasis and 11 health control subjects (n = 22 eyes) were prospectively recruited. ELISA for tear hexanoyllysine (HEL) and inflammatory cytokines, tear film break-up time tests, Schirmer test measurements, and fluorescein and rose bengal vital staining were performed. Conjunctival specimens obtained during surgery for conjunctivochalasis and cataract underwent immunohistochemical staining for HEL+8- OHdG, MMP-3, and MMP-9, and positively stained cells were counted. Transmission electron microscopy was also performed, with staining for elastic fibers in the conjunctival stroma. RESULTS. The mean tear stability and vital staining scores were significantly worse in the conjunctivochalasis patients than in the control subjects. The tear HEL and tear cytokine levels showed significantly higher values in eyes with conjunctivochalasis. IL-1β and IL-6 levels showed a significant correlation with corneal epithelial damage. IL-1β and TNFα showed a significant correlation with 8-OHdG-stained cell counts. Specimens from patients with conjunctivochalasis revealed a significantly higher number of cells positively stained for HEL, 8-OHdG, MMP-3, and MMP-9 than did specimens from age- and sex-matched control subjects. Transmission electron microscopy showed decreased intercellular cohesiveness, with the conjunctival stroma showing an accumulation of elastic fibers. CONCLUSIONS. Lipid and DNA oxidative stress were present in the conjunctiva. Increased tear inflammation seemed to coexist with loss of conjunctival epithelial cohesiveness and increased collagenolytic activity, which may explain the conjunctival laxity observed in patients with conjunctivochalasis. © Association for Research in Vision and Ophthalmology.

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Induction of corneal epithelium-like cells from cynomolgus monkey embryonic stem cells and their experimental transplantation to damaged cornea

Kumagai Yuta, Kurokawa Manae S., Ueno Hiroki, Kayama Maki, Tsubota Kazuo, Nakatsuji Norio, Kondo Yasushi, Ueno Satoki, Suzuki Noboru

Cornea 29 ( 4 ) 432 - 438 2010.04

ISSN 0277-3740

Purpose: We previously reported the successful transplantation of corneal epithelium-like cells derived from mouse embryonic stem (ES) cells onto injured mouse cornea. Here, we tested whether nonhuman primate ES cells have ability to differentiate into corneal epithelial cells and whether monkey ES cell-derived corneal epithelium-like cells were applicable for the experimental transplantation to damaged cornea. Methods: Monkey ES cells were cultivated on type IV collagen-coated dishes for various days to induce differentiation into corneal epithelium-like cells. The differentiation was evaluated by reverse transcription-polymerase chain reaction and immunostaining. The corneal epithelium-like cells were transplanted to the injured mouse cornea. Reconstitution of the corneal epithelium was evaluated by immunostaining. Results: The cells cultured on type IV collagen showed cobblestone like appearance resembling epithelial cells. They expressed messenger RNA of pax6, p63, E-cadherin, CD44, proliferating cell nuclear antigen, keratin 3, and keratin 12. Protein expressions of pax6, keratin 3/12, p63, proliferating cell nuclear antigen, E-cadherin, and CD44 were confirmed by immunostaining. When the corneal epithelium-like cells were transplanted, they adhered to the corneal stroma, leading to formation of multiple cell layers. The grafted cells were stained with anti-human nuclear protein antibody, which crossreacted with nuclei of monkey cells but not with those of mouse cells. They retained the expressions of keratin 3/12, E-cadherin, and CD44. Conclusions: We induced corneal epithelium-like cells from monkey ES cells with moderate efficiency. The cells were successfully transplanted onto the injured mouse cornea. This is the first demonstration that nonhuman primate ES cells were induced to differentiate into corneal epithelium-like cells, which were applicable for transplantation to an animal model of corneal injury. © 2010 Lippincott Williams & Wilkins.

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Erratum

Kawakita T., Kawashima M., Murat D., Tsubota K., Shimazaki J.

Eye 24 ( 4 ) 746 2010.04

ISSN 0950-222X

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Replacement of multifocal by monofocal intraocular lens in two cases for asthenopia and photophobia

Torii Hidemasa, Negishi Kazuno, Dogru Murato, Tsubota Kazuo

Japanese Journal of Clinical Ophthalmology 64 ( 4 ) 459 - 463 2010.04

ISSN 0370-5579

Purpose : To report two cases who received replacement of multifocal intraocular lens (IOL) by monofocal IOL due to postoperative dissatisfaction. Cases : A 75-year-old male received bilateral cataract surgery with implantation of diffractive multifocal IOL. Asthenopia and photophobia became worse after surgery. In accordance to his wish, the multifocal IOL was replaced by monofocal IOL 4 months later. His subjective complaints persisted but improved after treatment for essential blepharospasm. Another 52-year-old female received similar bilateral cataract surgery with implantation of refractive multifocal IOL. The lens was replaced by monofocal one due to photophobia 4 months later. Her subjective complaints persisted. Conclusion : These cases illustrate that replacement of multifocal by monofocal IOL due to dissatisfaction of patients need careful considerations.

Biconvex posterior chamber accommodating intraocular lens implantation after cataract surgery

Saiki Megumi, Negishi Kazuno, Dogru Murat, Yamaguchi Takefumi, Tsubota Kazuo

Journal of Cataract and Refractive Surgery 36 ( 4 ) 603 - 608 2010.04

ISSN 0886-3350

Purpose: To evaluate the long-term efficacy of a biconvex accommodating intraocular lens (IOL) to restore near visual performance. Setting: Department of Ophthalmology, Keio University Hospital, Tokyo, Japan. Methods: This 4-year study comprised eyes that had cataract surgery with implantation of a 1CU accommodating IOL. At all postoperative visits, the following were assessed: subjective spherical equivalent (SE) refraction; uncorrected (UDVA) and corrected (CDVA) distance visual acuities; uncorrected (UNVA), distance-corrected (DCNVA), and corrected (CNVA) near visual acuities; change in minimum additional (add) power (diopters) to attain CNVA; and accommodation amplitude. A questionnaire was administered at the last visit. Results: Twelve eyes of 8 cataract patients (mean age 59.0 years ± 18.4 [SD]) were evaluated. Postoperatively, the mean SE at 1 year and 4 years was significantly more hyperopic than at 1 month (P<.05). The mean UDVA increased significantly at 4 years compared with 1 month (P<.05). There were no significant changes in CDVA, UNVA, CNVA, and DCNVA throughout the follow-up period. The change in the minimum add power to attain CNVA and in the subjective and objective accommodation amplitudes also did not change significantly over time. Eighty-three percent of patients reported being satisfied with the results. Conclusion: Four years after implantation of an accommodating IOL, most patients had good vision and were satisfied, although the accommodation amplitude for near vision was not sufficient throughout the follow-up. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned. © 2010 ASCRS and ESCRS.

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The Efficacy, Sensitivity, and Specificity of In Vivo Laser Confocal Microscopy in the Diagnosis of Meibomian Gland Dysfunction

Ibrahim Osama M A, Matsumoto Yukihiro, Dogru Murat, Adan Enrique Sato, Wakamatsu Tais Hitomi, Goto Tateki, Negishi Kazuno, Tsubota Kazuo

Ophthalmology 117 ( 4 ) 665 - 672 2010.04

ISSN 0161-6420

Purpose: To evaluate the efficacy, sensitivity and specificity of confocal microscopy (CM) parameters: meibomian gland (MG) acinar longest diameter (MGALD), MG acinar shortest diameter (MGASD), inflammatory cell density (ICD), and MG acinar unit density (MGAUD) in the diagnosis of MG dysfunction (MGD). Design: Prospective, controlled, single-center study. Participants: Twenty MGD patients (9 males, 11 females; mean age, 63.5±16.5 years) and 26 age- and gender-matched control subjects (13 males, 13 females; mean age, 53.2±15.7 years) were recruited. Methods: All subjects underwent slit-lamp examinations, tear film break-up time (BUT) measurements, assessment of tear evaporation rate from the ocular surface (TEROS), vital stainings, Schirmer test, meibography, MG expressibility, and CM of the MG. Data were compared between the 2 groups using the Mann-Whitney and chi-square tests. Main Outcome Measures: The correlation between the clinical findings of tear functions, vital staining scores, and the 4 CM parameters were tested by Spearman's correlation coefficient by rank test. Receiver operating characteristic curve technique was used to evaluate the sensitivity, specificity, and cutoff values of CM parameters. Results: The mean tear film BUT, vital staining scores, TEROS values, MG expressibility, and MG dropout grades by meibography were significantly worse in MGD patients compared with controls (P<0.001). The mean values of the MGALD, MGASD, ICD, and MGAUD in MGD patients were significantly worse than those observed in the controls with CM. All CM parameters showed a strong, significant correlation with tear functions, ocular surface vital stainings, MG expressibility, and MG dropout grades. The cutoff values for MGALD, MGASD, ICD, and MGAUD in the diagnosis of MGD were 65 μm, 25 μm, 300 cells/mm², and 70 glands/mm², respectively. The sensitivity and specificity values of these parameters under these cutoff values were 90% and 81% for MGALD, 86% and 96% for MGASD, 100% and 100% for ICD, 81% and 81% for MGAUD. Conclusions: Confocal microscopy has the potential to diagnose the simple MGD with high sensitivity and specificity. The CM-based diagnostic parameters correlated significantly and strongly with the status of the ocular surface disease. Financial Disclosure(s): The authors have no proprietary or commercial interest in any of the materials discussed in this article. © 2010 American Academy of Ophthalmology.

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Mechanism of visual sensations experienced during pars plana vitrectomy under retrobulbar anesthesia

Sugisaka Eiko, Shinoda Kei, Sano Ronaldo Yuiti, Ishida Susumu, Imamura Yutaka, Ozawa Yoko, Shinoda Hajime, Suzuki Kotaro, Tsubota Kazuo, Inoue Makoto

Ophthalmologica 224 ( 2 ) 103 - 108 2010.02

ISSN 0030-3755

Purpose: To investigate the visual sensations experienced by patients during vitrectomy under retrobulbar anesthesia. Methods: 30 men and 45 women with a mean age of 65.3 ± 10.6 years underwent vitrectomy under retrobulbar anesthesia for macular disease. 28 eyes had an idiopathic epiretinal membrane, 13 had an idiopathic macular hole, 32 had macular edema (17 diabetic retinopathy and 15 retinal vein occlusion), and 2 had submacular hemorrhage. 49 patients with nonmacular disease underwent similar vitrectomy procedures and were used for comparison. An interview was conducted with the patient about his/her visual sensations during and within 3 h of the vitrectomy. Results: 70 (93.3%) of the patients reported seeing lights, 53 (70.7%) reported seeing colors, and 48 (64.0%) reported seeing movements or moving objects. Of the patients who reported seeing movements or moving objects, 44 (58.7%) reported seeing surgical instruments, and 5 (6.7%) saw the surgeon's fingers or hands. Patients with macular diseases tended to report more visual sensations than patients with nonmacular diseases. The patients' description and drawings appeared to arise mainly from the shadows cast by the intravitreal objects, and some patients perceived highly accurate details including the movements and color of the objects. Conclusions: Visual sensations are experienced by approximately 90% of the patients, and there may be a common mechanism by which patients perceive the intravitreal objects that are not focused on by the retina through the eye's optical system. © 2009 S. Karger AG, Basel.

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Conjunctival in vivo confocal scanning laser microscopy in patients with sjögren syndrome

Wakamatsu Tais Hitomi, Sato Enrique Adan, Matsumoto Yukihiro, Ibrahim Osama M A, Dogru Murat, Kaido Minako, Ishida Reiko, Tsubota Kazuo

Investigative Ophthalmology and Visual Science 51 ( 1 ) 144 - 150 2010.01

ISSN 0146-0404

PURPOSE. To demonstrate the conjunctival alterations in patients with Sjögren's (SSDE) and non-Sjögren's syndrome dry eye (NSSDE) using a new generation confocal microscope (HRTII/ RCM; Heidelberg Engineering, Heidelberg, Germany), in a prospective controlled study. METHODS. Twenty-eight right eyes of 28 patients with SSDE (28 women; mean age, 58.2 ± 14.3 years), 7 right eyes of patients with NSSDE (7 women; mean age, 66.1 ± 14.4 years), and 14 right eyes of 14 age- and sex-matched control subjects were studied. All subjects underwent the Schirmer test, tear film breakup time (BUT), vital staining, and confocal microscopy of the temporal bulbar conjunctiva. The density of conjunctival epithelial cells, epithelial microcysts, and conjunctival and corneal inflammatory infiltrates were also assessed. RESULTS. The tear quantity, stability, and vital staining scores were significantly worse in patients with SSDE or NSSDE than in control subjects (P < 0.001 and P < 0.05, respectively). Eyes of the patients with SSDE or NSSDE had a significantly higher density of conjunctival and corneal inflammatory infiltrates than did the control eyes (P < 0.001). Conjunctival inflammatory cell densities showed a negative correlation with tear stability and tear quantity and a positive correlation with the vital staining scores. Conjunctival epithelial cell densities were significantly lower in SSDE and NSSDE compared with control subjects (P < 0.05). The density of epithelial cysts was significantly higher in SS than in healthy control eyes (P < 0.001). CONCLUSIONS. Confocal scanning laser microscopy was an efficient and a noninvasive tool for the quantitative assessment of the conjunctival inflammation and epithelial cell densities as well as evaluation of conjunctival morphologic alterations, such as microcysts in patients with SSDE and NSSDE dry eye disease. © Association for Research in Vision and Ophthalmology.

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Subepithelial corneal fibrosis partially due to epithelial-mesenchymal transition of ocular surface epithelium

Kawashima Motoko, Kawakita Tetsuya, Higa Kazunari, Satake Yoshiyuki, Omoto Masahiro, Tsubota Kazuo, Shimmura Shigeto, Shimazaki Jun

Molecular Vision 16 2727 - 2732 2010

ISSN 1090-0535

Purpose: To determine whether epithelial-mesenchymal transition is involved in the development of corneal subepithelial fibrosis (pannus). Methods: Frozen samples of pannus tissue removed from human corneas with a diagnosis of total limbal stem cell deficiency were characterized by immunostaining for both epithelial and mesenchymal markers. We selected transformation-related protein 63 (p63) and pancytokeratin as epithelial markers and vimentin and α-smooth muscle actin (α-SMA) as mesenchymal markers. Immunostaining for β-catenin and E-cadherin was performed to determine wingless- Int (Wnt)-pathway activation. RT-PCR analysis was also performed on epithelial tissue obtained from pannus samples after dispase digestion. Results: Immunohistochemistry revealed strong nuclear expression of p63 and weak intercellular expression of E-cadherin in epithelial basal cells of pannus tissue. Furthermore, translocation of β-catenin from intercellular junctions to the nucleus and cytoplasm was also observed. Double-positive cells for both p63 and α-SMA were observed in the subepithelial stroma of pannus tissue, which was supported by RT-PCR and cytospin analysis. Conclusions: Epithelial-mesenchymal transition may be partially involved in the development of subepithelial corneal fibrosis due to total limbal stem cell deficiency. © 2010 Molecular Vision.

Topical tranilast for treatment of the early stage of mild dry eye associated with chronic GVHD

Ogawa Y., Dogru M., Uchino M., Tatematsu Y., Kamoi M., Yamamoto Y., Ogawa J., Ishida R., Kaido M., Hara S., Matsumoto Y., Kawakita T., Okamoto S., Tsubota K.

Bone Marrow Transplantation 45 ( 3 ) 565 - 569 2010

ISSN 0268-3369

Fibrosis and excessive extracellular matrix production are characteristic features of lacrimal gland chronic GVHD (cGVHD). Tranilast (n-3,4-anthoranilic acid), used for fibrotic skin disease, inhibits transforming growth factor-Β-induced matrix production. We conducted a non-randomized study comparing 8 patients (five men, three women; median age, 47 years) given topical tranilast with 10 patients (three men, seven women; median age, 37 years) receiving therapy with topical artificial tears, sodium hyaluronate and vitamin A for mild ocular cGVHD. The tranilast group instilled topical tranilast and artificial tears q.i.d., beginning the day of dry eye diagnosis. The ocular surface and tear dynamics of each patient were evaluated before hematopoietic stem cell transplant, at the onset of dry eye and after 3 months of treatment. At 3 months, the scores of the Rose Bengal test and Schirmer test with nasal stimulation had significantly improved in the tranilast group compared with that in the control group (P<0.05). Furthermore, although five control patients (50) developed severe dry eye within the treatment period, only one tranilast-treated patient (12.5) did; the rest still had mild dry eye (P<0.05). These results suggest the hypothesis that topical tranilast may effectively retard the progression of mild dry eye associated with cGVHD. © 2010 Macmillan Publishers Limited.

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Lacrimal hypofunction as a new mechanism of dry eye in visual display terminal users

Nakamura Shigeru, Kinoshita Shigeru, Yokoi Norihiko, Ogawa Yoko, Shibuya Michiko, Nakashima Hideo, Hisamura Ryuji, Imada Toshihiro, Imagawa Tomohiro, Uehara Masato, Shibuya Izumi, Dogru Murat, Ward Samantha, Tsubota Kazuo

PLoS ONE 5 ( 6 ) 2010

Background: Dry eye has shown a marked increase due to visual display terminal (VDT) use. It remains unclear whether reduced blinking while focusing can have a direct deleterious impact on the lacrimal gland function. To address this issue that potentially affects the life quality, we conducted a large-scale epidemiological study of VDT users and an animal study. Methodology/Principal Findings: Cross sectional survey carried out in Japan. A total of 1025 office workers who use VDT were enrolled. The association between VDT work duration and changes in tear film status, precorneal tear stability, lipid layer status and tear secretion were analyzed. For the animal model study, the rat VDT user model, placing rats onto a balance swing in combination with exposure to an evaporative environment was used to analyze lacrimal gland function. There was no positive relationship between VDT working duration and change in tear film stability and lipid layer status. The odds ratio for decrease in Schirmer score, index of tear secretion, were significantly increased with VDT working year (P = 0.012) and time (P = 0.005). The rat VDT user model, showed chronic reduction of tear secretion and was accompanied by an impairment of the lacrimal gland function and morphology. This dysfunction was recovered when rats were moved to resting conditions without the swing. Conclusions/Significance: These data suggest that lacrimal gland hypofunction is associated with VDT use and may be a critical mechanism for VDT-associated dry eye. We believe this to be the first mechanistic link to the pathogenesis of dry eye in office workers. © 2010 Nakamura et al.

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Serum free fatty acids levels not associated with normal tension glaucoma

Yuki Kenya, Kimura Itaru, Tsubota Kazuo

Clinical Ophthalmology 4 ( 1 ) 91 - 94 2010

ISSN 1177-5467

Purpose: To determine the free fatty acid levels in the sera of patients with normal-tension glaucoma and compare it with that of normal controls. Methods: Forty-four consecutive patients with newly diagnosed normal-tension glaucoma and forty-four age and gender matched controls were evaluated. The type and level of fatty acids in the sera were measured by gas chromatography (Model GC17A; Shimazu, Kyoto, Japan). Twenty-four fatty acids were identified from 12:0 to 24:1. The values were compared between the normal-tension glaucoma and control groups by Mann-Whitney U tests. Results: No statistically significant difference was found in the levels of any free fatty acids between the normal-tension glaucoma group and control group. Conclusion: No significant association was found in the serum free fatty acids levels including docosahexaenoic acid and eicosapentaenoic acid between normal-tension glaucoma patients and controls. © 2010 Yuki et al, publisher and licensee Dove Medical Press Ltd.

Reduced-serum vitamin C and increased uric acid levels in normal-tension glaucoma

Yuki Kenya, Murat Dogru, Kimura Itaru, Ohtake Yuichiro, Tsubota Kazuo

Graefe's Archive for Clinical and Experimental Ophthalmology 248 ( 2 ) 243 - 248 2010

ISSN 0721-832X

Background: Vitamin A, B₉, C, E, and uric acid are well-known antioxidants and may prevent age-related eye disorders. The aim of the present study was to investigate the levels of antioxidant vitamins, A, B₉, C, E, and antioxidative substance, uric acid in the serum of Japanese patients with normal-tension glaucoma and compare the results with normal controls. Methods: All subjects with suspicion of primary open-angle glaucoma who came to the glaucoma subspeciality clinic of Keio University Hospital were enrolled in this study. Sixty patients (28 males, 32 females; mean age ± standard deviation: 59.9 ± 9.8 years) with newly diagnosed primary open-angle glaucoma patients were consecutively enrolled in this study. After the diagnosis of primary open-angle glaucoma, the patients underwent 24-h IOP measurements. Forty-seven newly diagnosed consecutive normal-tension glaucoma patients (18 males, 29 females; mean age ± standard deviation: 59.5 ± 10.2 years) were enrolled in this study. The control subjects were recruited from subjects who came to the clinic for annual refractive check-up. The 44 consecutive control subjects of the current study, (16 males, 28 females; 62.7 ± 14.8 years) did not have any ocular diseases. The serum levels of vitamins A, B₉, C, E, and uric acid were measured. The values were compared between the normal-tension glaucoma and control groups by the Mann-Whitney U test. Results: Serum levels of vitamin C were significantly lower in normal-tension glaucoma patients than in normal healthy controls (P = 0.04; normal-tension glaucoma; 4.6 ± 4.0 μg/ml control; 6.3 ± 3.9 μg/ml). Uric acid level was significantly higher in normal-tension glaucoma patients than in controls (P = 0.01; normal-tension glaucoma; 5.8 ± 1.5 mg/dl control; 4.9 ± 1.4 mg/dl). No statistically significant difference was seen in vitamin A (P = 0.41; normal-tension glaucoma; 82.1 ± 26.7 μg/dl control; 77.1 ± 30.1 μg/dl), B₉ (P = 0.37; normal-tension glaucoma; 8.7 ± 4.3 ng/ml control; 8.0 ± 3.1 ng/ml)and E (P = 0.83; normal-tension glaucoma; 1.5 ± 0.6 control; 1.5 ± 0.6) levels between normal-tension glaucoma and control groups. Conclusion: Normal-tension glaucoma patients had lower serum levels of vitamin C and increased levels of uric acid. These observations may pave the way for possible alternative treatment for normal-tension glaucoma. © 2009 Springer-Verlag.

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Evaluation of lipid oxidative stress status and inflammation in atopic ocular surface disease

Wakamatsu Tais H., Dogru Murat, Ayako Igarashi, Takano Yoji, Matsumoto Yukihiro, Ibrahim Osama M A, Okada Naoko, Satake Yoshiyuki, Fukagawa Kazumi, Shimazaki Jun, Tsubota Kazuo, Fujishima Hiroshi

Molecular Vision 16 2465 - 2475 2010

ISSN 1090-0535

Background: Although the oxidative stress status in atopic skin disease has been reported to be elevated, there are still no studies related to the status of oxidative stress in atopic ocular surface disease. The purpose of this study was to evaluate the ocular surface lipid oxidative stress status and inflammation in atopic keratoconjunctivitis (AKC) patients and normal subjects. Methods: Twenty eight eyes of 14 patients (9 males, 5 females) with AKC and 18 eyes of 9 age and sex matched (4 males and 5 females) normal healthy controls were examined in this prospective study. The severity of atopic dermatitis (AD) was scored by the SCORing Atopic Dermatitis (SCORAD) index. All subjects underwent Schirmer test, tear film break up time (BUT), fluorescein/Rose Bengal stainings, tear collection, and brush cytology from the upper palpebral conjunctiva. The brush cytology samples were stained with Diff-Quik for differentiation of inflammatory cells and immunohistochemistry (IHC) staining with HEL (hexanoyl-lysine) and 4-HNE (4-hydroxy-2-nonenal) to study lipid oxidation. HEL and cytokine (interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-10 (IL-10), tumor necrosis factoralpha (TNF-α), interferon-gamma (IFN-γ)) levels were measured by enzyme-linked immunosorbent assay (ELISA) from tear samples of AKC patients and control subjects. Toluidine Blue and IHC staining with HEL, 4-HNE and cluster of differentiation 45 (CD45) were performed on papillary samples of AKC patients. This study was conducted in compliance with the "Declaration of Helsinki." Results: The tear stability and vital staining scores were significantly worse in eyes of AKC patients (p<0.05) compared to the controls. Inflammatory cells and positively stained conjunctival epithelial cells for HEL and 4-HNE showed a significant elevation in brush cytology samples of AKC patients. Significantly higher levels of HEL and cytokines were detected in tears of AKC patients compared to controls. Papillary specimens also revealed many CD45 inflammatory cells as well as many cells positively stained with HEL and 4-HNE in IHC. A strong significant linear positive correlation between conjunctival inflammation and epithelial lipid oxidative stress status was observed. Conjunctival lipid oxidative stress also correlated strongly with tear HEL levels and epithelial damage scores. Conclusions: The ocular surface disease in AKC was characterized by marked tear instability, ocular surface epithelial damage, increase in inflammatory infiltrates and presence of increased lipid oxidation. © 2010 Molecular Vision.

The effect of topical amiloride eye drops on tear quantity in rabbits

Hara Shuya, Hazama Akihiro, Miyake Masao, Kojima Takashi, Sasaki Yasumasa, Shimazaki Jun, Dogru Murat, Tsubota Kazuo

Molecular Vision 16 2279 - 2285 2010

ISSN 1090-0535

Purpose: To investigate the presence of epithelial sodium channels (ENaC) in rabbit and human conjunctival epithelium and to test the effects of topical amiloride, a potassium-sparing diuretic that blocks the ENaC, on tear quantity in rabbits. Methods: Both healthy normal human and rabbit conjunctival tissues underwent immunohistochemistry staining for ENaC-α and γ subunits as well as for reverse transcription polymerase chain reaction (RT-PCR) for detection of ENaC- α and ENaC-γ subunit mRNA expression. Rabbits were instilled topical amiloride eye drops and tear function tests were performed before and after instillations. Results: Immunohistochemical staining for ENaC-α subunit in all rabbit eyes showed positive staining in apical and basal conjunctival epithelial cells. Human conjunctival epithelia revealed positive staining with ENaC-α antibody especially in the apical and basal layers. Immunohistochemistry staining with ENaC- γ antibody also revealed positive staining of the conjnunctival epithelial cells especially in the basal layers. The ENaC-α mRNA was detected in samples from healthy white rabbit conjunctival epithelia, and ENaC-α and ENaC- γ mRNAs were detected in samples from healthy human conjunctival epithelia. The mean tear quantity showed a significant increase at 15 and 30 min compared to the pre-instillation value in eyes assigned to amiloride eye drops. The mean tear quantity at 15 and 30 min was significantly higher in the amiloride group compared to the control eyes. Conclusions: Topical amiloride application appears to increase the quantity of preocular tears owing to inhibition of conjunctival epithelial sodium channels. © 2010 Molecular Vision.

Anti-aging in ophthalmology.

Tsubota Kazuo

Ophthalmic research 44 ( 3 ) 145 2010

Pattern-reversal visual-evoked potential in patients with occult macular dystrophy

Hanazono Gen, Ohde Hisao, Shinoda Kei, Tsunoda Kazushige, Tsubota Kazuo, Miyake Yozo

Clinical Ophthalmology 4 ( 1 ) 1515 - 1520 2010

ISSN 1177-5467

Purpose: Occult macular dystrophy (OMD) is a hereditary retinal disease characterized by a normal fundus, normal full-field electroretinograms (ERGs), progressive decrease of visual acuity, and abnormal focal macular ERGs. The purpose of this study was to report pattern-reversal visual-evoked potential (pVEPs) findings in OMD patients. Patients and method: The pVEPs recorded from four patients with OMD (aged 42-61 years; 2 men and 2 women) were reviewed. The visual acuities ranged from 20/200 to 20/30. The amplitudes of the N-75 and P-100 (P2 amplitude) and the latency of the N-75 components (N1 latency) were analyzed. Results: The mean (±SD) P2 amplitude was 2.7 ± 1.9 μV for the 5′, 4.8 ± 2.9 μV for the 10′, 3.2 ± 2.1 μV for the 20′, and 4.4 ± 3.5 μV for the 40′ checkerboard stimuli. The N1 latency was 122.2 ± 6.4 ms for the 5′, 105.0 ± 11.5 ms for the 10′, 97.7 ± 10.0 ms for the 20′, and 91.0 ± 13.7 ms for the 40′ checkerboard stimuli. The mean P2 amplitude was reduced and the N1 latency was delayed in comparison with the laboratory standard for the Keio University Hospital. Conclusions: The delayed latency and reduced amplitude suggest a major contribution of the central cone pathway to the pVEPs. © 2010 Hanazono et al.

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Selenoprotein P controls oxidative stress in cornea

Higuchi Akihiro, Takahashi Kazuhiko, Hirashima Masaki, Kawakita Tetsuya, Tsubota Kazuo

PLoS ONE 5 ( 3 ) 2010

The ocular surface is always attacked by oxidative stress, and cornea epithelial cells are supposed to have their own recovery system against oxidative stress. Therefore we hypothesized that tears supply key molecules for preventing oxidative stress in cornea. The potential target key molecule we focused is selenoprotein P (SeP). SeP is a carrier of selenium, which is an essential trace element for many animals, for oxidative stress metabolism in the organism, and was extremely expressed in lacrimal gland. An experiment was performed with SeP eye drops in a rat dry eye model, prepared by removing the lacrimal glands. The anticipated improvement in corneal dry eye index and the suppression of oxidative stress markers were observed in SeP eye drop group. Furthermore, the concentration of SeP was significantly higher in dry eye patients compared with normal volunteers. Collectively, we concluded that tear SeP is a key molecule to protect the ocular surface cells against environmental oxidative stress. © 2010 Higuchi et al.

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Eicosapentaenoic acid suppresses ocular inflammation in endotoxin-induced uveitis

Suzuki Misa, Noda Kousuke, Kubota Shunsuke, Hirasawa Manabu, Ozawa Yoko, Tsubota Kazuo, Mizuki Nobuhisa, Ishida Susumu

Molecular Vision 16 1382 - 1388 2010

ISSN 1090-0535

Purpose: To investigate the effect of eicosapentaenoic acid (EPA) on acute ocular inflammation in an animal model of endotoxin-induced uveitis (EIU). Methods: C57Bl/6 mice (6-week-old males) were orally treated with EPA at a dose of 50 mg/kg/day for 5 days. EIU was then induced in the animals by intraperitoneal injection of 160 μg lipopolysaccharide (LPS). Twenty-four hours after LPS injection, leukocyte adhesion to the retinal vasculature was evaluated by the concanavalin A lectin perfusion-labeling technique, and leukocyte infiltration into the vitreous cavity was quantified. Furthermore, the protein levels of monocyte chemotactic protein (MCP)-1, interleukin (IL)-6, intercellular adhesion molecule-1 and phospholyrated nuclear factor (NF)-κB p65 in the retina and retinal pigment epithelium (RPE)-choroid complex were examined by enzyme-linked immunosorbent assay (ELISA). Results: At 24 h after LPS injection, the EIU animals treated with oral EPA administration showed a significant decrease in leukocyte adhesion to the retinal vessels by 43.4% (p<0.01) and leukocyte infiltration into the vitreous cavity by 49.2% (p<0.05). In addition, EPA significantly reduced the protein levels of MCP-1 and IL-6 in the retina and the RPE-choroid complex. Furthermore, phosphorylation of NF-κB was suppressed by EPA treatment. Conclusions: Our data suggest that EPA inhibits multiple inflammatory molecules in vivo. EPA may become a novel strategy in the prevention and/or treatment of ocular inflammatory diseases. © 2010 Molecular Vision.

Evaluation of ocular surface glycocalyx using lectin-conjugated fluorescein

Mochizuki Hiroshi, Fukui Masaki, Hatou Shin, Yamada Masakazu, Tsubota Kazuo

Clinical Ophthalmology 4 ( 1 ) 925 - 930 2010

ISSN 1177-5467

Purpose: A glycocalyx plays important roles in the ocular surface epithelium, but there is no direct simple method to evaluate ocular surface glycocalyx. We tested a wheat germ agglutinin conjugate of fluorescein (F-WGA) as a marker to demonstrate ocular surface glycocalyx in vivo. Methods: After a 5% F-WGA solution was applied to the eyes of eight healthy volunteers, fluorescent intensities of the central cornea and bulbar conjunctiva were measured by fluorophotometry. A 10% fluorescein-conjugated dextran solution served as the control. Changes in fluorescent intensities of the ocular surface following a challenge with 5% N-acetyl cysteine, a conventional mucolytic agent, were tested in the second experiment. Saline instillation served as a control. Results: The ocular surface was diffusely stained by F-WGA. Breakup of the precorneal tear film was not apparent, possibly because F-WGA was bound to the glycocalyx of the ocular surface epithelium. F-WGA fluorescent intensities were high in the superior, nasal, and inferior regions of the bulbar conjunctiva and low in the temporal conjunctiva and cornea. No such regional differences were observed with fluorescein-conjugated dextran. F-WGA fluorescent intensities decreased significantly with N-acetyl cysteine instillation, whereas they were not affected by saline instillation. Conclusion: The fluorophotometric method may be used to evaluate the glycocalyx quantitatively in the ocular surface in vivo. © 2010 Mochizuki et al.

Comparison of LASIK with the OPDCAT or OATz algorithm using the NIDEK EC-5000CXII excimer laser

Hori-Komai Yoshiko, Toda Ikuko, Yamamoto Takahiro, Tsubota Kazuo

Journal of Refractive Surgery 26 ( 6 ) 411 - 422 2010

ISSN 1081-597X

PURPOSE: To compare refractive outcomes, higher order aberrations, visual quality, and patient satisfaction between aspheric and whole-eye wavefront aspheric LASIK algorithms. METHODS: Two hundred seventy-four eyes of 152 patients undergoing LASIK for myopia and myopic astigmatism were divided into two groups: eyes that underwent treatment using either the OPD-guided customized aspheric treatment (OPDCAT) or optimized aspheric treatment zone (OATz). Both groups were subdivided into two groups based on preoperative manifest refraction spherical equivalent (MRSE) <-6.00 diopters (D) and eyes with MRSE ≥-6.00 D. Both groups were additionally subdivided into eyes with preoperative ocular higher order aberrations <0.40 μm and eyes with higher order aberrations ≥0.40 μm. A P value <.05 was considered statistically significant. RESULTS: At 3 months postoperatively, 88.3% (242/274) of eyes were available for follow-up. Postoperatively, 91.4% of eyes in the OPDCAT group and 90.6% of eyes in the OATz group were within 0.50 D. No difference in refractive outcomes and patient satisfaction among groups or subgroups was noted (P>.05). A significantly less change in asphericity (less oblate) was noted for the OPDCAT group (0.31±0.30) compared with the OATz group (0.51±0.35) (P<.05). A lower induction of aberrations in the OPDCAT group compared with the OATz group was noted (P<.05). Mesopic contrast sensitivity was significantly higher for the OPDCAT groups and subgroups (P<.05). CONCLUSIONS: Refractive outcomes between groups or subgroups were equivalent. A significantly lower induction of higher order aberrations and less change in asphericity in the OPDCAT group was noted. The OPDCAT algorithm was more likely to maintain mesopic contrast sensitivity. Copyright © SLACK Incorporated.

Access to Document (DOI)

The Application of in vivo laser scanning confocal microscopy as a tool of conjunctival in vivo cytology in the diagnosis of dry eye ocular surface disease

Kojima Takashi, Matsumoto Yukihiro, Dogru Murat, Tsubota Kazuo

Molecular Vision 16 2457 - 2464 2010

ISSN 1090-0535

Purpose: To evaluate the applicability of in vivo laser scanning confocal microscopy as a tool of conjunctival cytology in a prospective case-control study. Methods: Nineteen right eyes of 19 Sjogren's syndrome dry eye patients (19 females; mean age: 55.8±15 years), and 18 right eyes of 18 normal healthy control subjects (12 females and 6 males; mean age: 50.8±14 years) were evaluated in this study. The eyes were analyzed by the Heidelberg retina tomography (HRTII)/Rostock cornea module (RCM). Ocular surface and tear function tests including vital stainings (fluorescein and Rose Bengal), Schirmer test, tear film break up time (BUT), and conjunctival impression cytology were performed. After obtaining the confocal microscopy images, the mean individual epithelial cell area (MIECA), and nucleocytoplasmic (N/C) ratio were analyzed. The correlation between confocal microscopy and impression cytology parameters was also investigated. Results: The BUT, Schirmer test values, vital staining scores and squamous metaplasia grades in impression cytology were significantly worse in dry eye patients compared to controls (p<0.0001). The MIECA and the mean N/C ratios were worse in dry eye subjects compared to controls both in impression cytology and in vivo confocal microscopy (p<0.0001) with no significant differences between these parameters when the two examination techniques were compared. The MIECA and N/C ratio in conjunctival impression cytology showed significant correlation with the corresponding confocal microscopy parameters (MIECA, r²:0.557; N/C, r²:0.765). Conclusions: Laser scanning confocal microscopy seems to be an efficient non-invasive tool in the evaluation of phenotypic alterations of the conjunctival epithelium in dry eye disease. N/C ratio and MIECA appear to be two promising and new parameters of in vivo confocal cytology in the assessment of the ocular surface in dry eye disease. © 2010 Molecular Vision.

Inflammatory cells in brush cytology samples correlate with the severity of corneal lesions in atopic keratoconjunctivitis.

'Takano Y, Fukagawa K, Dogru M, Asano-Kato N, Tsubota K, Fujishima H.'

Br J Ophthalmol 88 ( 12 ) 1504-1505 2004.12

Research paper (scientific journal), Joint Work

INTACS before or after laser in situ keratomileusis: correction of thin corneas with moderately high myopia.

'Ito M, Arai H, Fukumoto T, Toda I, Tsubota K.'

J Refract Surg 20 ( 6 ) 818-822 2004.12

Research paper (scientific journal), Joint Work

Induction of epithelial progenitors in vitro from mouse embryonic stem cells and application for reconstruction of damaged cornea in mice.

'Homma R, Yoshikawa H, Takeno M, Kurokawa MS, Masuda C, Takada E, Tsubota K, Ueno S, Suzuki N.'

Invest Ophthalmol Vis Sci 45 ( 12 ) 4320-4326 2004.12

Research paper (scientific journal), Joint Work

Effect of preoperative tear function on early functional visual acuity after laser in situ keratomileusis.

'Tanaka M, Takano Y, Dogru M, Toda I, Asano-Kato N, Komai-Hori Y, Tsubota K.'

J Cataract Refract Surg 30 ( 11 ) 2311-2315 2004.11

Research paper (scientific journal), Joint Work

Dramatic healing of an allergic corneal ulcer persistent for 6 months by amniotic membrane patching in a patient with atopic keratoconjunctivitis: a case report.

Takano Y, Fukagawa K, Miyake-Kashima M, Tanaka M, Asano-Kato N, Dogru M, Tsubota K, Fujishima H.

Cornea 23 ( 7 ) 723-725 2004.10

Research paper (scientific journal), Joint Work

Lidocaine tape (Penles) reduces the pain of botulinum toxin injection for Meige syndrome.

'Onguchi T, Takano Y, Dogru M, Ono M, Tsubota K.'

Am J Ophthalmol 138 ( 4 ) 654-655 2004.10

Research paper (scientific journal), Joint Work

ドライアイ治療2004アップデート,角膜診療における患者さんの満足度アップ

坪田一男

ドライアイ研究会「角膜診療クオリティアップ1dayセミナー」 2004.09

Single Work

Ocular surface changes with applanation contact lens and coupling fluid use after argon laser photocoagulation in noninsulin-dependent diabetes mellitus.

Dogru M, Kaderli B, Gelisken O, Yucel A, Avci R, Goto E, Shimmura S, Shimazaki J, Tsubota K.

Am J Ophthalmol 138 ( 3 ) 381-388 2004.09

Research paper (scientific journal), Joint Work

Role of the endothelial pump in flap adhesion after laser in situ keratomileusis.

Bissen-Miyajima H, Nakamura K, Kaido M, Shimmura S, Tsubota K.

J Cataract Refract Surg 30 ( 9 ) 1989-1992 2004.09

Research paper (scientific journal), Joint Work

「アンチエイジング注目のEBM」

坪田一男

ＭCL抗加齢医学の実際2004−新しい医療を学ぶ２日間− 2004.09

Single Work

目から考えるアンチエイジング

坪田一男

日経プラスワンフォーラム 2004.09

Single Work

βブロッカー点眼と防腐剤が涙液・眼表面に及ぼす影響

'石岡みさき, 島崎潤, 八木幸子, 坪田一男'

臨床眼科(0370-5579) 58 ( 8 ) 1437-1440 2004.08

Research paper (scientific journal), Joint Work

Increased tear evaporation in a patient with ectrodactyly-ectodermal dysplasia-clefting syndrome.

Matsumoto Y, Dogru M, Goto E, Endo K, Tsubota K.

Jpn J Ophthalmol 48 ( 4 ) 372-375 2004.08

Research paper (scientific journal), Joint Work

Efficacy of core vitrectomy preceding triple corneal procedure.

Konomi K, Shimazaki J, Shimmura S, Akabane N, Goto E, Tsubota K.

Br J Ophthalmol 88 ( 8 ) 1023-1025 2004.08

Research paper (scientific journal), Joint Work

The relation of conjunctival and corneal findings in severe ocular allergies.

Tanaka M, Dogru M, Takano Y, Miyake-Kashima M, Asano-Kato N, Fukagawa K, Tsubota K, Fujishima H.

Cornea 23 ( 5 ) 464-467 2004.07

Research paper (scientific journal), Joint Work

Efficacy and safety of international donor sharing: a single-center, case-controlled study on corneal transplantation.

Shimazaki J, Shinozaki N, Shimmura S, Holland EJ, Tsubota K.

Transplantation 78 ( 2 ) 216-220 2004.07

Research paper (scientific journal), Joint Work

Therapeutic effect of cevimeline on dry eye in patients with Sjogren's syndrome: a randomized, double-blind clinical study.

Ono M, Takamura E, Shinozaki K, Tsumura T, Hamano T, Yagi Y, Tsubota K.

Am J Ophthalmol 138 ( 1 ) 39250 2004.07

Research paper (scientific journal), Joint Work

Quantitative evaluation of eyelid elasticity using the cutometer SEM575 and its clinical application in assessing the efficacy of tacrolimus ointment treatment in eyelid atopic dermatitis.

Kawakita T, Takano Y, Asano-Kato N, Tanaka M, Dogru M, Goto E, Tsubota K, Takahashi S, Fukagawa K, Fujishima H.

Cornea 23 ( 5 ) 468-471 2004.07

Research paper (scientific journal), Joint Work

Ocular surface treatment before laser in situ keratomileusis in patients with severe dry eye.

Toda I, Asano-Kato N, Hori-Komai Y, Tsubota K.

J Refract Surg 20 ( 3 ) 270-275 2004.06

Research paper (scientific journal), Joint Work

Autologous serum application in the treatment of neurotrophic keratopathy.

Matsumoto Y, Dogru M, Goto E, Ohashi Y, Kojima T, Ishida R, Tsubota K.

Ophthalmology 111 ( 6 ) 1115-1120 2004.06

Research paper (scientific journal), Joint Work

Risk factors and retreatment results of intraoperative flap complications in LASIK.

Ito M, Hori-Komai Y, Toda I, Tsubota K.

J Cataract Refract Surg 30 ( 6 ) 1240-1247 2004.06

Research paper (scientific journal), Joint Work

LASIK直後に明らかな原因なくフラップ偏位・剥離を起こした3症例

'石田玲子, 池田宏一郎, 加藤直子, 戸田郁子, 堀好子, 坪田一男'

あたらしい眼科(0910-1810) 21 ( 6 ) 832-835 2004.06

Research paper (scientific journal), Joint Work

角膜移植ハイリスク症例に対するシクロスポリン全身投与の効果と安全性

'大本雅弘, 榛村重人, ムラト・ドール, 島崎潤, 坪田一男'

臨床眼科(0370-5579) 58 ( 5 ) 773-776 2004.05

Research paper (scientific journal), Joint Work

Tear function and ocular surface findings in premature and term babies.

Dogru M, Karakaya H, Baykara M, Ozmen A, Koksal N, Goto E, Matsumoto Y, Kojima T, Shimazaki J, Tsubota K.

Ophthalmology 111 ( 5 ) 901-905 2004.05

Research paper (scientific journal), Joint Work

Punctal occlusion in the management of chronic Stevens-Johnson syndrome.

Kaido M, Goto E, Dogru M, Tsubota K.

Ophthalmology 111 ( 5 ) 895-900 2004.05

Research paper (scientific journal), Joint Work

A new noninvasive tear stability analysis system for the assessment of dry eyes.

Kojima T, Ishida R, Dogru M, Goto E, Takano Y, Matsumoto Y, Kaido M, Ohashi Y, Tsubota K.

Invest Ophthalmol Vis Sci 45 ( 5 ) 1369-1374 2004.05

Research paper (scientific journal), Joint Work

Same-size donor corneas for myopic keratoconus.

Shimmura S, Ando M, Ishioka M, Shimazaki J, Tsubota K.

Cornea 23 ( 4 ) 345-359 2004.05

Research paper (scientific journal), Joint Work

A comparative evaluation of the efficacy of intraoperative mitomycin C use after the excision of cobblestone-like papillae in severe atopic and vernal keratoconjunctivitis.

Tanaka M, Takano Y, Dogru M, Fukagawa K, Asano-Kato N, Tsubota K, Fujishima H.

Cornea 23 ( 4 ) 326-329 2004.05

Research paper (scientific journal), Joint Work

調節可能眼内レンズを用いた白内障手術の成績

'本田理恵, 村戸ドール, 藤島浩, 戸田郁子, 荒井宏幸, 坪田一男'

臨床眼科(0370-5579) 45 ( 6 ) 901-905 2004.04

Research paper (scientific journal), Joint Work

Detection of neutrophils in late-onset interface inflammation associated with flap injury after laser in situ keratomileusis.

Asano-Kato N, Toda I, Fukumoto T, Asai H, Tsubota K.

Cornea 23 ( 3 ) 306-310 2004.04

Research paper (scientific journal), Joint Work

Early visual results with the rollable ThinOptX intraocular lens.

Dogru M, Honda R, Omoto M, Fujishima H, Yagi Y, Tsubota K, Kojima T, Matsuyama M, Nishijima S, Yagi Y.

J Cataract Refract Surg 30 ( 3 ) 558-565 2004.03

Research paper (scientific journal), Joint Work

Salivary flow and its relationship to oral signs and symptoms in patients with dry eyes.

Koseki M, Maki Y, Matsukubo T, Ohashi Y, Tsubota K.

Oral Dis 10 ( 2 ) 75-80 2004.03

Research paper (scientific journal), Joint Work

UV dose-dependent caspase activation in a corneal epithelial cell line.

Shimmura S, Tadano K, Tsubota K.

Curr Eye Res 28 ( 2 ) 85-92 2004.02

Research paper (scientific journal), Joint Work

Risk factors for development of diffuse lamellar keratitis after laser in situ keratomileusis.

Noda-Tsuruya T, Toda I, Asano-Kato N, Hori-Komai Y, Fukumoto T, Tsubota K.

J Refract Surg 20 ( 1 ) 72-75 2004.02

Research paper (scientific journal), Joint Work

再生角膜の作成及び移植に関する研究　研究経過報告

'石川達也, 榛村重人, 島崎潤, 臼井あけ美, 篠崎尚史, 深川和己, 吉田悟, 斉藤一郎, 藤島浩, 高野洋之, 坪田一男'

歯科学報(0037-3710) 104 ( 1 ) 39278 2004.02

Research paper (scientific journal), Joint Work

Donor source affects the outcome of ocular surface reconstruction in chemical or thermal burns of the cornea.

Shimazaki J, Shimmura S, Tsubota K.

Ophthalmology 111 ( 1 ) 38-44 2004.01

Research paper (scientific journal), Joint Work

Clinical Evaluation of the Accuracy of Intraocular Pressure Measurement by Tono-Pen XL in Eyes with Amniotic Membrane Patching.

Onguchi T, Takano Y, Dogru M, Ono M, Tsubota K, Shimazaki J

Am J Ophthalmol in press ( - ) 2004

Research paper (scientific journal), Joint Work

Selective suppression of pathological, but not physiological, retinal neovascularization by blocking angiotensin II type 1 receptor.

Nagai N, Noda K, Urano T, Kubota Y, Shinoda H, Koto T, Shinoda K, Inoue M, Shiomi T, Ikeda E, Tsubota K, Suda T, Oike Y, Ishida S.

Invest Ophthalmol Vis Sci ( in press ) 2004

Research paper (scientific journal), Joint Work

Revised Japanese criteria for Sjögren’s syndrome(1999): availability and validity.

Fujibayashi T, Sugai S, Miyasaka N, Hayashi Y, Tsubota K

Mod Rhematol 14 ( - ) 425-434 2004

Research paper (scientific journal), Joint Work

Electrolysis for Corneal Opacities in a Young Patient with Superficial Variant of Granular Corneal Dystrophy.

Kamoi M, Mashima Y, Kawashima M, Tsubota K.

Am J Ophthalmol in press ( - ) 2004

Research paper (scientific journal), Joint Work

Improved functional visual acuity after punctal occlusion in dry eye patients

Goto E, Yagi Y, Kaido M, Matsumoto Y, Konomi K, Tsubota K

American Journal of Ophthalmology 135 ( 5 ) 704-705 2003.05

Research paper (scientific journal), Joint Work

Tear evaporation dynamics in normal subjects and subjects with obstructive meibomian gland dysfunction

Goto E, Endo K, Suzuki A, Fujikura Y, Matsumoto Y, Tsubota K

Investigative Ophthalmology and Visual Science 44 ( 2 ) 533-539 2003.05

Research paper (scientific journal), Joint Work

Related Articles, Links Visualization of gaseous monoxide reception by soluble guanylate cyclase in the rat retina

Kajimura Mayumi, Shimoyama Masaru, Tsuyama Shingo, Suzuki Tsuneharu, Kozaki Shinji, Takenaka Shingo, Tsubota Kazuo, Oguchi Yoshihisa, Suematsu Makoto

FASEB journal 17 ( 3 ) 506-508 2003.03

Research paper (scientific journal), Joint Work

Treatment of non-inflamed obstructive meibomian gland dysfunction by an infrared warm compression device

Goto E, Monden Y, Takano Y, Mori A, Shimmura S, Shimazaki J, Tsubota K

British Journal of Ophthalmology 86 1403-1407 2002.12

Research paper (scientific journal), Joint Work

Low-concentration homogenized castor oil eye drops for noninflamed obstructive meibomian gland dysfunction

Goto E, Shimazaki J, Monden Y, Takano Y, Yagi Y, Shimmura S, Tsubota K

Ophthalmology 109 ( 11 ) 2030-2035 2002.11

Research paper (scientific journal), Joint Work

Dry eye assiciated with chronic graft-versus-host disease

Ogawa Yoko, Masataka Kuwana, Yamazaki Kazuto, Mashima Yukihiko, Okamoto Shinichiro, Tsubota Kazuo, Oguchi Yoshihisa, Yutaka Kawakami

Advances in experimental medicine and biology 506 ( Pt B ) 1041-1045 2002.10

Research paper (scientific journal), Joint Work

ステロイドパルス療法が有効であった抗ラミニン5瘢痕性類天疱瘡の1例

Inada Sachiko, Kobayashi Seiichirou, Ooyama Manabu, Tanikawa Akiko, Amagai Masayuki, Matsui Chihiro, Morohashi Masaaki, Tsubota Kazuo

臨床皮膚科 56 ( 9 ) 731-734 2002.08

Research paper (scientific journal), Joint Work

Impaired functional visual acuity of dry eye patients

Goto E, Yagi Y, Matsumoto Y, Tsubota K

American Journal of Ophthalmology 133 ( 2 ) 181-186 2002.02

Research paper (scientific journal), Joint Work

Successful treatment of dry eye in two patients with chronic graft-versus-host disease with systemic administration of FK506 and corticosteroids

Ogawa Y, Okamoto S, Kuwana M, Mori T, Watanabe R, Nakajima T, Yamada M, Mashima Y, Tsubota K, Oguchi Y

Cornea 20 ( 4 ) 430-434 2001.05

Research paper (scientific journal), Joint Work

Quantitative evaluation of atopic blepharitis by scoring of eyelid conditions and measuring the water content of the skin and evaporation from the eyelid surface

Asano-Kato N, Fukagawa K, Tsubota K, Urayama K, Takahashi S, Fujishima H

Cornea 20 ( 3 ) 255-259 2001.04

Research paper (scientific journal), Joint Work

Successful treatment of dry eye in two patients with chronic graft-versus-host disease with systemic administration of FK506 and corticosteroids: case reports

Ogawa Yoko, Okamoto Shinichiro, Kuwana Masataka, Mori Takehiko, Watanabe Reiko, Nakajima Takeshi, Yamada Masakazu, Mashima Yukihiko, Tsubota Kazuo, Oguchi Yoshihisa

Cornea 20 ( 4 ) 430-434 2001.04

Research paper (scientific journal), Joint Work

Lack of evidence for an increased microchimerism in circulation of patients with Sjogrens syndrome

Toda Ikuko, Kuwana Masataka, Tsubota Kazuo, Kawakami Yutaka

Annals of Rheumatic Diseases 60 ( 3 ) 248-253 2001.03

Research paper (scientific journal), Joint Work

Intact corneal epithelium is essential for the prevention of stromal haze after laser assisted in situ keratomileusis.

Nakamura K, Kurosaka D, Bissen-Miyajima H, Tsubota K

Br J Ophthalmol 85 ( 2 ) 209-213 2001.02

Research paper (scientific journal), Joint Work

A significant role of stromal fibroblasts in rapidly progressive dry eye in patients with chronic GVHD

Ogawa Yoko, Yamazaki Kazuto, Kuwana Masataka, Mashima Yukihiko, Nakamura Yu, Ishida Susumu, Toda Ikuko, Oguchi Yoshihisa, Tsubota Kazuo, Okamoto Shinichiro, Kawakami Yutaka

Investigative Ophthalmology & Visual Science 42 ( 1 ) 111-119 2001.01

Research paper (scientific journal), Joint Work

A significant role of stromal fibroblasts in rapidly progressive dry eye in patients with chronic GVHD

Ogawa Yoko, Yamazaki Kazuto, Kuwana Masataka, Mashima Yukihiko, Nakamura Yu, Ishida Susumu, Toda Ikuko, Oguchi Yoshihisa, Tsubota Kazuo, Okamoto Shinichiro, Kawakami Yutaka

Investigative Ophthalmology & Visual Science 42 ( 1 ) 111-119 2001.01

Research paper (scientific journal), Joint Work

Dry eye after haematopoietic stem cell transplantation

Ogawa Yoko, Okamoto Shinichiro, Wakui Masatoshi, Watanabe Reiko, Yamada Masakazu, Yoshino Mami, Ono Masafumi, Yang Hao-Yung, Mashima Yukihiko, Oguchi Yoshihisa, Ikeda Yukihiko, Tsubota Kazuo

British Journal of Ophthalmology 83 ( 10 ) 1125-1130 1999.10

Research paper (scientific journal), Joint Work

Dry eye after haematopoietic stem cell transplantation

Ogawa Yoko, Okamoto Shinichiro, Wakui Masatoshi, Watanabe Reiko, Yamada Masakazu, Yoshino Mami, Ono Masafumi, Yang Hao-Yung, Mashima Yukihiko, Oguchi Yoshihisa, Ikeda Yasuo, Tsubota Kazuo

British Journal of Ophthalmology 83 ( 10 ) 1125-1130 1999.10

Research paper (scientific journal), Joint Work

Il-4 regulates chemokine production induced by TNF-a in keratocytes and corneal epithelial cells

Takano Yoji, Fukagawa Kazumi, Shimmura Shigeto, Tsubota Kazuo, Oguchi Yoshihisa, Saito Hirohisa

British Journal of Ophthalmology 84 ( 9 ) 1074-1076 1999.09

Research paper (scientific journal), Joint Work

Thermal effect on corneal incisions with different phacoemulsification ultrasonic tips

Bissen-miyajima H, Shimmura S, Tsubota K

Journal of Cataract and Refractive Surgery 25 60-64 1999

Research paper (scientific journal), Joint Work

Treatment of dry eye by autologus serum application in Sjogren's syndrome

Tsubota K, Goto E, Fujita T, Ono M, Inoue H, Saito I, Shimmura S

British Journal of Ophthalmology 83 390-395 1999

Research paper (scientific journal), Joint Work

Efficacy and safety of infrared warming of the lids

Mori A, Oguchi Y, Goto E, Nakamori K, Ohtsuki T, Egami F, Shimazaki J, Tsubota K

Cornea 18 188-193 1999

Research paper (scientific journal), Joint Work

Pirfenidone induces intercellular adhesion molecule-1 (ICAM-1)down-regulation on cultured human synovial fibroblasts

Kaneko M, Inoue H, Nakazawa R, Azuma N, Suzuki M, Yamauchi S, Margolin S, Tsubota K, Saito I

Clinical and Experimental Immunololgy 113 72-76 1998

Research paper (scientific journal), Joint Work

Intra-versus post-operative suture adjustment following penetrating keratoplasty

Shimazaki J, Shimmura S, Tsubota K

Cornea 17 590-594 1998

Research paper (scientific journal), Joint Work

Terrien's marginal degeneration associated with erythema elevatum diutinum

Shimazaki J, Yang H-y, Shimmura S, Tsubota K

Cornea 17 342-344 1998

Research paper (scientific journal), Joint Work

Reoxygenation injury in a cultured corneal epithelial cell line protected by the uptake of lactoferrin

Shimmura S, Shimoyama M, Hojo M, Urayama K, Tsubota K

Investigative Ophthalmology and Visual Science 39 1346-1351 1998

Research paper (scientific journal), Joint Work

Real-time telemedicine in the clinical assessment of the ocular surface

Shimmura S, Shinozaki S, Fukagawa K, Shimazaki J, Tsubota K

American Journal of Ophthalmology 125 388-390 1998

Research paper (scientific journal), Joint Work

T new surgical technique for deep lamellar keratoplasty with single running suture adjustment

Tsubota K, Kaido M, Monden Y, Satake Y, Bissen-miyajima H, Shimazaki J

American Journal of Ophthamology 126 1-8 1998

Research paper (scientific journal), Joint Work

全層角膜移植術後のシクロスポリン点眼の有用性

Matsumoto Yukihiro, Oyama Mitsuko, Satake Yoshiyuki, Shimazaki Jun, Tsubota Kazuo

臨眼 52 957-962 1998

Research paper (scientific journal), Joint Work

春季カタルにおけるBrush Cytology検体中Eosinophil Cationic Proteinと角膜障害の相関

Nakamura Maki, Fukagawa Kazumi, Saito Hirohisa, Tsubota Kazuo, Oguchi Yoshihisa

あたらしい眼科 15 693-696 1998

Research paper (scientific journal), Joint Work

Allergic conjunctivities as a risk factor for regression and haze after photorefractive keratectomy

Yang H-y, Fujishima H, Toda I, Bissen-miyajima H, Shimazaki J, Tsubota K

American Journal of Ophthalmology 125 54-58 1998

Research paper (scientific journal), Joint Work

角膜移植のための角膜組織保存法

Konomi Kenji, Shinozaki Naofumi, Tsubota Kazuo

Organ Biology 5 27-32 1998

Research paper (scientific journal), Joint Work

Lecithin-bound superoxide dismutase in the prevention of neutrophil-induced damage of corneal tissue

Matsumoto K, Shimmura S, Goto E, Saito K, Takeuchi T, Miyajima S, Negi A, Tsubota K

Investigative Ophthalmololgy and Visual Science 39 30-35 1998

Research paper (scientific journal), Joint Work

Meibomian gland dysfunction in patients with Sjogren syndrome

Shimazaki J, Goto E, Ono M, Shimmura S, Tsubota K

Ophthamology 105 1485-1488 1998

Research paper (scientific journal), Joint Work

Viscosity-dependent fluid dymanics of eyedrops on the ocular surface

Shimmura S, Goto E, Shimazaki J, Tsubota K

American Journal of Ophthalmology 125 386-388 1998

Research paper (scientific journal), Joint Work

エキシマレーザー屈折矯正手術後のコントラスト視力

Takano Youji, Yan Haoyun, BiッSen Miyajima Hiroko, Shimazaki Jun, Tsubota Kazuo, Itou Seiji, Katsumi Osamu

日眼紀 48 ( 10 ) 1183-1185 1997.10

Research paper (scientific journal), Joint Work

Subthreshold UV Light-induced peroxide formation in cultured corneal epithelial cells

Shimmura Shigeto, Suematsu Makoto, Shimoyama M, Tsubota Kazuo, Oguchi Y, Ishimura Yuzuru

Exp Eye Res 63 ( 1 ) 519-526 1996.11

Research paper (scientific journal), Joint Work

Scanning electric microscopic observation of basal cells following corneal epithelial abrasion

Yamada Masakazu, Mashima Yukihiko, Tsubota Kazuo

Eye 10 ( 5 ) 569-574 1996.09

Research paper (scientific journal), Joint Work

オクラーサーフェスと皮膚

Tsubota Kazuo, Shimizu Hiroshi

あたらしい眼科 13 ( 3 ) 319-320 1996.03

Research paper (scientific journal), Joint Work

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