Details of a Researcher - Tsubota, Kazuo

Papers - Tsubota, Kazuo

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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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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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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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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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