Atsushi Shiraishi

Ehime University, Matuyama, Ehime, Japan

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Publications (77)192.75 Total impact

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    ABSTRACT: Objective: To characterize the clinical profile of patients with recurrent subconjunctival hemorrhages (SCHs) and evaluate the effect of conjunctivochalasis (CCh) surgery on disease recurrences. Methods: Three hundred and sixty-two patients with SCHs (mean age, 56.4±16.0 years) were enrolled in this multicenter epidemiologic study. The severity of CCh, lifestyle at the time of SCH onset, and the frequency of previous SCHs were compared. Thirty-eight patients with 2 or more episodes of SCHs (mean age, 68.2±8.9 years) underwent surgery for CCh. The effectiveness of surgery was evaluated by comparing the frequency of SCH preoperatively and postoperatively. Results: Patients with three or more recurrent SCHs showed a significantly (P=0.003) higher grade of CCh and tended to be engaged in activities that require visual concentration, such as watching a visual display terminal, knitting, reading, and driving. More than 80% of eyes that underwent surgery to CCh showed no recurrence of the hemorrhages, and the frequency of SCH significantly (P<0.0001) decreased postoperatively. Conclusions: Moderate or severe CCh and activities that may cause dry eye can be considered to be risk factors for recurrent SCHs. Surgery to treat CCh is a useful option for patients with frequent recurrences of SCHs.
    Eye & Contact Lens Science & Clinical Practice 08/2015; DOI:10.1097/ICL.0000000000000139 · 1.47 Impact Factor
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    ABSTRACT: To determine the relationship between eyelid pressure during blinking and the fluorescein staining of the cornea and conjunctiva in dry eye patients. Cross-sectional prospective study. The pressure of the upper and lower eyelids was measured with a specially designed blepahro-tensiometer in 130 eyes of 65 dry eye patients (D group) and in 58 eyes of 31 normal controls (N group). The correlations between the location and degree of ocular surface staining scores and the eyelid pressure were calculated for the D group. The pressure of the upper and lower eyelids were significantly higher in the D group than in the N group (upper P <0.0001, lower P = 0.0040). The lower eyelid pressure was significantly correlated with the ocular surface staining scores for the inferior cornea (r = 0.19, P = 0.0307) and conjunctiva ((r = 0.19, P = 0.0252). The significant correlation between the eyelid pressure and the ocular surface staining suggests that mechanical friction on the ocular surface by the eyelids may be one of the factors that affects the fluorescein staining of the inferior ocular surface. Copyright © 2015 Elsevier Inc. All rights reserved.
    American Journal of Ophthalmology 07/2015; 160(4). DOI:10.1016/j.ajo.2015.06.028 · 3.87 Impact Factor
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    ABSTRACT: To determine whether eyelid pressure is involved in the development of lid-wiper epitheliopathy (LWE). Study 1: The eyelid pressure was measured with a blepharo-tensiometer, and the degree of LWE was assessed in 79 eyes of 43 non-contact lens (CL) wearers. Study 2: The movements of the eyelids and displacement of the eyes during spontaneous blinking were photographed with a high-speed camera. The eyelid pressure was also measured in 34 normal eyes of 19 non-CL wearers who were not part of Study 1. Study 1: Upper-LWE was detected in 24 of 79 eyes (30.4%), and no significant difference was detected in the eyelid pressure between any grade of upper-LWE. Lower-LWE was detected in 41 of 79 eyes (51.9%), and the eyelid pressure (27.9 ± 2.8 mmHg) in eyes with grade 3 LWE was significantly higher than that with grade 0 LWE (19.7 ± 1.3 mmHg; p < 0.05). Study 2: The lower eyelid pressure was significantly correlated with the length of the horizontal movement of the lower eyelids (p < 0.05) and also with the degree of posterior movement of the eye globe (p < 0.05). The higher pressure from the eyelid may be one of the causes for the development of lower-LWE.
    Current eye research 03/2015; DOI:10.3109/02713683.2015.1009636 · 1.64 Impact Factor
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    ABSTRACT: Interactions between stromal and epithelial cells play important roles in the development, homeostasis, and pathological conditions of the cornea. Soluble cytokines are critical factors in stromal–epithelial interactions, and growth factors secreted from corneal stromal cells contribute to the regulation of proliferation and differentiation of corneal epithelial cells (CECs). However, the manner in which the expression of growth factors is regulated in stromal cells has not been completely determined. To study stromal–epithelial cell interactions, we used an organotypic culture model. Human or rabbit CECs (HCECs or RCECs) were cultured on amniotic membranes placed on human corneal fibroblasts (HCFs) embedded in a collagen gel. The properties of the organotypic culture were examined by hematoxylin–eosin staining and immunofluorescence. In the organotypic culture, HCECs or RCECs were stratified into two–three layers after five days and five–seven layers after nine days. However, stratification was not observed when the HCECs were seeded on a collagen gel without fibroblasts. K3/K12 were expressed on day 9. The HCF-embedded collagen gels were collected on days 3, 5, or 9 after seeding the RCECs, and mRNA expression of growth factors FGF7, HGF, NGF, EGF, TGF-α, SCF, TGF-β1, TGF-β2, and TGF-β3 were quantified by real-time PCR. mRNA expression of the growth factors in HCFs cultured with RCECs were compared with those cultured without RCECs, as well as in monolayer cultures. mRNA expression of TGF-α was markedly increased in HCFs cultured with RCECs. However, mRNA expression of the TGF-β family was suppressed in HCFs cultured with RCECs. Principal component analysis revealed that mRNA expression of the growth factors in HCFs were generally similar when they were cultured with RCECs. In organotypic cultures, the morphological changes in the CECs and the expression patterns of the growth factors in the stromal cells clearly demonstrated stromal–epithelial cell interactions, and the results suggest that stromal cells and epithelial cells may act in concert in the cornea.
    Experimental Eye Research 02/2015; 135. DOI:10.1016/j.exer.2015.02.009 · 2.71 Impact Factor
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    ABSTRACT: Miraflow is a cleaner for soft contact lens which contains 20% isopropyl alcohol. The purpose of this study was to determine the activity against Acanthamoeba trophozoites and cysts for Miraflow. In addition, to determine the activity of combined Miraflow and multipurpose solutions (MPSs) against Acanthamoeba cysts. Two simulated-use studies were conducted. The significance in the log reduction in the number of trophozoites and cysts of A. castellanii strains ATCC 50514 and ATCC 50370 or A. polyphaga ATCC 30461 after exposure to Miraflow alone was determined by the Spearman-Karber method. To examine the activity against Acanthamoeba of combined Miraflow and an MPS, the log reduction in the number of cysts after a 1-min exposure to Miraflow followed by a 4-hr exposure to MPS (ReNu fresh) was also determined. Short-time exposure of 30 sec to Miraflow demonstrated activity against the Acanthamoeba trophozoites. However, a 1-min treatment was only relatively effective (1.1 log reduction) against the cysts of A. castellanii ATCC 50514, but no statistically significant reduction was observed for the cysts of the other 2 strains. The combined use with Miraflow and MPS demonstrated activity against the cysts, and a 3.0, 1.0, or 1.5 log reduction in the numbers was obtained for A. castellanii ATCC 50514, A. castellanii ATCC 50370, and A. polyphaga ATCC 30461, respectively. Exposure to combined Miraflow and MPS resulted in reductions in the number of Acanthamoeba cysts.
    Eye & Contact Lens Science & Clinical Practice 01/2015; Publish Ahead of Print(4). DOI:10.1097/ICL.0000000000000112 · 1.47 Impact Factor
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    ABSTRACT: PURPOSE. To test our hypothesis about whether there is water migration in the horizontal corneal plane and investigate its developmental mechanism. METHODS. A fluorescein solution was intrastromally injected into normal and edematous corneas of rabbits, and the movement of the fluorescein solution was observed and recorded over time. RESULTS. In normal corneas, the water flow was characterized by a swirling movement in the stroma. The fluorescein solution ultimately spread and occupied the entire cornea, indicating horizontal intracorneal swirling of water. In contrast, when the corneal endothelia were injured by intracameral injection of a preservative to create corneal edema, no water migration occurred, suggesting that the integrity of the corneal endothelial function is essential for water migration. The water migration stopped with injection of a sodium-potassium pump inhibitor, indicating that the enzyme is necessary for physiologic water migration in the cornea. With recovery of corneal endothelial function, the water migration began, and focal edema remained in the periphery with no water migration in this edematous area. CONCLUSIONS. We report for the first time the presence of horizontal water migration in the cornea in a swirling pattern, i.e., intracorneal swirling migration of water, generated by the pump function in the corneal endothelial cells, which may supplement the conventional concept of development of corneal edema in the vertical plane. This dynamic water circulatory system may be involved in increasing the efficiency of the water transfer in the entire cornea. Copyright © 2014 by Association for Research in Vision and Ophthalmology.
    Investigative Ophthalmology &amp Visual Science 11/2014; 55(12). DOI:10.1167/iovs.14-14762 · 3.40 Impact Factor
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    ABSTRACT: Background Beauveria bassiana is an entomopathogenic fungus and is a rare cause of keratitis. We present a case of fungal keratitis caused by B. bassiana that was diagnosed by in vivo confocal microscopy and in vitro corneal cultures. In addition, we determined the temperature- and drug-sensitivities of the isolated strain of B. bassiana. Case presentation A 59-year-old Japanese man with a 2-month history of keratitis was examined by slit-lamp biomicroscopy, in vivo confocal microscopy, and histology and cultures of corneal scrapings. The corneal scrapings were used to determine the minimal inhibitory concentrations of different antifungal drugs and also to determine the temperature-sensitivity. In vivo confocal microscopy and histological examinations showed filamentous fungal keratitis. The characteristics of the fungal growth indicated that the keratitis was caused by B. bassiana. The keratitis responded poorly to systemic and topical voriconazole and to natamycin ointment. However, it was resolved after changing the natamycin to micafungin combined with surgical debridement. The isolated strain was sensitive to itraconazole, miconazole, micafungin, voriconazole, and resistant to flucytosine and fluconazole. It was moderately sensitive to amphotericin B, and natamycin. After 7 days in culture, the isolate grew small white colonies at 25°C, very small colonies at 35°C and 37°C. Conclusion The drug-sensitivity and temperature-sensitivity profiles of B. bassiana should be helpful in the treatment of B. bassiana keratitis. Therapeutic surgery may be helpful for mycotic keratitis poorly responsive to medical therapy alone.
    BMC Research Notes 09/2014; 7(1):677. DOI:10.1186/1756-0500-7-677
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    ABSTRACT: Aims The purpose of this study is to elucidate the clinical manifestations and the current treatment status of cytomegalovirus (CMV) endotheliitis via a large case series obtained from a national survey conducted in Japan. Methods The Japan Corneal Endotheliitis Study Group proposed diagnostic criteria for CMV endotheliitis based on a viral examination by PCR of aqueous humour, in combination with clinical manifestations. A national survey was then retrospectively conducted among 1160 members of the Japan Cornea Society. The study reviewed the patient profiles, clinical manifestations, and treatment modalities of individuals who met the diagnostic criteria for CMV endotheliitis. Results The study included 109 eyes of 106 patients. Mean patient age was 66.9±10.9 years (85 males (80.2%), 21 females (19.8%)). Patients were commonly diagnosed with anterior uveitis and ocular hypertension prior to confirmation of CMV endotheliitis. Coin-shaped lesions were observed in 70.6%, and linear keratic precipitates in 8.3% of the patients, respectively. 95% of cases were treated with anti-CMV drugs. Conclusions CMV endotheliitis is most common in middle-aged and elderly men. CMV endotheliitis should be suspected when patients present with corneal endotheliitis involving coin-shaped lesions accompanied by anterior uveitis and ocular hypertension.
    British Journal of Ophthalmology 07/2014; 99(1). DOI:10.1136/bjophthalmol-2013-304625 · 2.98 Impact Factor
  • Atsushi Shiraishi · Masahiko Yamaguchi · Yuichi Ohashi
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    ABSTRACT: Objective: To report lid-wiper epitheliopathy (LWE)-like staining at the lower eyelid margin (lower-LWE) and to determine the prevalence of LWE (upper-LWE) and lower-LWE in contact lens (CL) wearers and non-CL wearers. Methods: Four hundred forty-three eyes of 229 non-CL wearers, 405 eyes of 208 soft CL wearers, and 135 eyes of 71 rigid gas permeable CL wearers were studied. Lissamine green and fluorescein (FL) staining were used to assess the degree of LWEs, tear break-up time (BUT), and cornea-conjunctival staining (FL-S). The correlations between the prevalence of LWEs and the other factors were evaluated. Results: The prevalence of lower-LWE was significantly higher (39.5%) than upper-LWE in non-CL wearers (upper-LWE; 12.0%; P<0.001). The prevalence of both upper- and lower-LWE were significantly correlated with age and FL-S, but not sex and BUT, in non-CL wearers. The prevalence of both upper- and lower-LWE was significantly higher in younger than older subjects (P<0.001). Upper- and lower-LWE were detected in a higher percentage of CL wearers than in non-CL wearers (P<0.001). Conclusion: Our results indicate that examination of the lower eyelid margin would be preferable to that of the upper eyelid margin in studies of LWE.
    Eye & Contact Lens Science & Clinical Practice 06/2014; 40(4). DOI:10.1097/ICL.0000000000000040 · 1.47 Impact Factor
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    ABSTRACT: PurposeTo investigate the changes in the tear flow velocities caused by ageing.Methods Ninety-nine subjects (41 men, mean age 48.3 ± 20.7 years) were recruited from the Department of Ophthalmology of the Ehime University Hospital. None of the subjects had serious abnormalities of the external surface of the eye. The Krehbiel flow of tears was determined by 40-μm polymethylmethacrylate (PMMA) beads suspended in a fluorescein sodium solution (PPF). The movement of the beads was video recorded through a slit-lamp during normal blinking. The flow of the beads was determined with a Motion analyzer®software (KEYENCE Co., Osaka, Japan). The velocity of the beads in young age, 20–40 years, middle age, 41–60 years and old age, ≥61 years, groups was determined.ResultsThe equation describing the velocity (mm/second) of the PMMA particles as a function of age in the lower tear meniscus measured in the direction of the lacrimal punctum was Y = 2.49−0.04X, where Y = velocity and X = age (r2 = 0.214; p < 0.0001). For the upper meniscus, the equation was Y = 4.83−0.05X (r2 = 0.195, p < 0.0001). The average velocity was 0.70 ± 1.66 mm/second in the lower and 2.16 ± 1.93 mm/second in the upper tear meniscus (p < 0.0001). The particle velocity decreased significantly with increasing age, but no significant difference between the male and female groups except for the lower tear meniscus when all subjects were analysed.Conclusion The PPF technique is a simple method of examining Krehbiel flow of tears and may be used for evaluating functional nasolacrimal duct obstruction quantitatively.
    Acta ophthalmologica 06/2014; 92(8). DOI:10.1111/aos.12444 · 2.84 Impact Factor
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    ABSTRACT: Purpose: To evaluate ocular surface temperature in assessing the efficacy of anti-allergic eye drops. Methods: Thirteen asymptomatic patients (24.7 ± 2.8 years) with proven seasonal allergic conjunctivitis due to cedar pollen were studied. A 0.025% levocabastine ophthalmic suspension was instilled in one eye (levocabastine eye) and artificial tears in the other eye (artificial tear eye) in a masked fashion 10 min prior to a conjunctival allergen challenge (CAC). Then, a drop of cedar pollen solution was dropped into the conjunctival sac to induce the allergic reaction. The surface temperature of the inferior bulbar conjunctiva was measured before and 30 min after the CAC with a newly developed non-contact ocular surface thermographer (OST). The degree of conjunctival injection and chemosis was also determined by slit-lamp biomicroscopy. The changes in the symptoms were evaluated by a questionnaire. Results: After the CAC, the temperature increased by 0.67 ± 0.10°C in the artificial tear eyes but by only 0.21 ± 0.06°C in the levocabastine eyes (p < 0.05). The score for conjunctival injection was 1.38 ± 0.24 and the chemosis score was 0.85 ± 0.25 for the artificial tear eyes and 0.62 ± 0.27 and 0.08 ± 0.08 in the levocabastine eyes (p < 0.01). The temperature increase was significantly correlated with the conjunctival injection scores (r = 0.63; p < 0.001). Conclusion: The significant correlation of the conjunctival surface temperature with the severity of the conjunctival allergic reaction indicates that the temperature measured by the OST can be used to objectively evaluate the efficacy of topical anti-allergic agents.
    Ophthalmic Research 03/2014; 51(3):161-166. DOI:10.1159/000357105 · 1.42 Impact Factor
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    ABSTRACT: Purpose: To investigate the differing characteristics of limbal niche cells (LNCs) and limbal stromal cells (LSCs) in the maintenance of limbal epithelial stem/progenitor cells in the cornea. Methods: Limbal niche cells were obtained from direct dissection of the human corneal limbus, and LSCs were obtained from explant cultures of limbal stromal tissues under the same culture conditions. The resulting cultures were examined for their ability to support the growth of limbal stem/progenitor cells in colony-forming capacity, stratified epithelial cell sheet formation, maintenance of limbal epithelial stem/progenitor cell characteristics, and gene expression levels of factors that supported the limbal epithelial stem/progenitor cells. Results: The colony-forming efficiency of limbal epithelial stem/progenitor cells in the LNC group (6.57 ± 1.54%) was significantly higher than that in the LSC group (1.43 ± 0.47%). The epithelial cell sheets in the LNC group stratified into four or five layers compared with two or three stratified layers in the LSC group. Staining of both the colonies and the epithelial cell sheets in the LNC group showed a higher intensity of the limbal stem cell marker ΔNp63 than in the LSC group. Moreover, reverse transcription polymerase chain reaction analysis revealed that compared with the common expression of EGF and so on, the LNCs showed a higher expression level of E-cadherin and a lower expression level of neurotrophin-3 (NT3) than the LSCs. Conclusions: LNCs have a different role compared to LSCs in their ability to support epithelial stem/progenitor cells and epithelial cellular sheet formation.
    Investigative ophthalmology & visual science 01/2014; 55(3). DOI:10.1167/iovs.13-13698 · 3.40 Impact Factor
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    ABSTRACT: To develop a side-view imaging technique for observing the dynamic behavior of posterior chamber structures (PCSs) in porcine eyes which mimics closed-eye cataract surgery in humans. Enucleated porcine eyes were placed into liquid nitrogen for 5 seconds and immediately bisected at about a 45-degree angle to the equatorial plane. The anterior portion was attached firmly to a glass slide with superglue and sprinkled with wheat flour. Phacoemulsification and aspiration (PEA) was performed as in humans on 10 consecutive porcine eyes. The movements of the PCSs were monitored through the glass slide with a high-resolution video camera set below the cut surface of the eye. The intraocular pressure (IOP) was monitored during the surgery. The highest IOP, operation time, and volume of irrigation fluid of 10 whole eyes were compared to that obtained from the bisected eyes glued to a glass slide. In a second set of experiments, the strength of the seal between the bisected eye and the glass slide was tested in three sets of eyes: 1) frozen eye fixed with superglue with wheat flour for 3 min; 2) frozen eye fixed with superglue for 3 min; and 3) non-frozen eye fixed with superglue for 30 min. The highest IOP that led to a disruption of the seal was compared among the three groups. PEA was successfully performed on 9 of 10 (90%) eyes with the movements of the PCSs clearly observed. The average maximum intraocular pressure of the 9 bisected eyes was 55.8 +/- 4.7 mmHg and that for the 10 unbisected eyes was 55.3 +/- 5.0 mmHg (P = 0.650). The frozen eye fixed with superglue in combination with wheat flour (Group 1) had the strongest sealing strength with an average IOP at the breaking point of 117.3 +/- 36.2 mmHg. Our side-view imaging technique can be used to evaluate the changes of the PCSs during intraocular surgery and for surgical training of new residents.
    BMC Ophthalmology 09/2013; 13(1):47. DOI:10.1186/1471-2415-13-47 · 1.02 Impact Factor
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    ABSTRACT: To describe a new method of measuring early phase tear clearance by anterior segment optical coherence tomography (AS-OCT). Sixty normal subjects were divided into a young group (30 subjects; 29.6 ± 7.2 years) and an elder group (30 subjects; 71.4 ± 10.8 years). AS-OCT (CASIA SS-1000, Tomey, Japan) with customized software was used to record the tear meniscus at the centre of the lower eyelid. Five microlitres of lukewarm saline solution was dropped into the lower conjunctival sac, and an image of the tear meniscus was obtained immediately and again 30 seconds after natural blinking. The tear meniscus height (TMH) and tear meniscus area (TMA) were measured in the AS-OCT images, and the percentage decrease in the TMH and TMA was used as a measure of the tear clearance. Correlations between tear clearance and clinical features including degree of conjunctivochalasis, degree of protrusion of inferior lacrimal punctum, distance of lacrimal punctum from the Marx line and fluorescein clearance rates were also determined in another healthy population consisting of 30 subjects. The OCT tear clearance rate was 35.2 ± 11% for TMH and 28.1 ± 12.4% for TMA in the young group, and 12.4 ± 7.3% and 6.2 ± 9.1%, respectively in the elder group. The differences were significant for both the TMH (p = 0.017) and the TMA (p = 0.024). The OCT-determined tear clearance was positively correlated with the fluorescein clearance rate, and negatively correlated with the distance between the lacrimal punctum and Marx line, degree of conjunctivochalasis and degree of lacrimal punctum protrusion. AS-OCT can be used as a rapid, non-invasive and quantitative method of determining the early phase tear clearance rate in a normal healthy population.
    Acta ophthalmologica 09/2013; 92(2). DOI:10.1111/aos.12260 · 2.84 Impact Factor
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    ABSTRACT: We report a patient who, based on the clinical manifestations, was originally diagnosed as having Chandler's syndrome and later developed varicella-zoster virus (VZV) DNA-positive anterior uveitis. The patient with Chandler's syndrome who manifested anterior uveitis underwent a complete ophthalmologic examination. Polymerase chain reaction (PCR) was used to amplify the viral DNA in the aqueous humor to determine the cause of the intraocular inflammation. Slit-lamp biomicroscopy showed focal iris atrophy and peripheral anterior synechiae (PAS); specular microscopy of the corneal endothelium disclosed the hammered-silver appearance. Based on these clinical findings, we diagnosed this patient as having Chandler's syndrome. During the follow-up period, however, the inflammatory cells suddenly appeared in the anterior chamber with formation of keratic precipitates and an increased intraocular pressure (IOP). VZV DNA was displayed in the aqueous humor by PCR. Based upon the diagnosis of VZV anterior uveitis, corticosteroids and acyclovir were given topically and systemically. The inflammation subsided with these medications; however, trabeculectomy was finally needed to control the IOP due to PAS progression. The coincidence of VZV anterior uveitis with Chandler's syndrome may constitute an implication for the possible viral etiology of iridocorneal endothelial syndrome.
    Case Reports in Ophthalmology 09/2013; 4(3):274-8. DOI:10.1159/000357239
  • Xiaodong Zheng · Tomoko Goto · Atsushi Shiraishi · Yuichi Ohashi
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    ABSTRACT: To compare 3 ocular lubricants containing sodium hyaluronate (SH), carboxymethylcellulose (CMC), and hydroxypropyl methylcellulose (HPMC) for their ability to enhance water retention and to protect human corneal epithelial cells (HCECs) from dehydration. Experiments were performed using 0.1% and 0.3% solutions of the 3 lubricants diluted in Milli-Q water for the water retention assays and in DMEM/F12 culture medium for the cell viability assays. Five milliliters of each of the lubricants was dropped onto a filter paper, and the paper was kept in an open container at 25°C and a humidity of 36% to 38%. The weight of the paper was measured hourly for 4 hours. In the second set of experiment, cultured HCECs were exposed to the test lubricants for 60 minutes, and the lubricants were removed. Cells were then exposed to room air for up to 60 minutes. Cells were then incubated with a vital dye, and the absorption of the reduced form of the dye was measured. The cell survival rate was compared for the 3 lubricants. Filter papers moistened with both 0.1% and 0.3% SH were significantly heavier than those moistened with CMC and HPMC at all time points. The survival rate of HCECs was significantly higher at most times with 0.1% and 0.3% SH than with CMC and HPMC solutions. The effects of CMC were not significantly different from those of HPMC. These findings indicate that SH is significantly better for water retention and protection of HCECs from dehydration than HPMC and CMC.
    Cornea 07/2013; 32(9). DOI:10.1097/ICO.0b013e31829cfd44 · 2.04 Impact Factor
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    ABSTRACT: Because human corneal endothelial cells do not proliferate once the endothelial monolayer is formed, corneal wound healing is thought to be mediated by cell enlargement or migration rather than proliferation. However, the cellular mechanisms involved in corneal wound healing have not been fully determined. Because transforming growth factor-β(2) (TGF-β(2)) isoform is present in high concentrations in normal human aqueous humor, it may play a role in human corneal endothelial cell wound healing. The purpose of this study was to determine the effect of TGF-β(2) on the proliferation and migration of cultured human corneal endothelial cells (HCECs). To achieve this, we first examined the effect of TGF-β(2) on the wound closure rate in an in vitro HCEC wound healing model. However, unexpectedly TGF-β(2) had no effect on the wound closure rate in this model. Therefore, a real-time cell electronic sensing (RT-CES) system and the BrdU incorporation assay were used to determine the effect of recombinant TGF-β(2) (0.1-10 ng/ml) on cultured HCEC proliferation during in vitro wound healing. The specificity of this effect was confirmed by adding the TGF-β receptor I kinase inhibitor. TGF-β(2) inhibited the proliferation of HCECs in a dose dependent way and was blocked by TGF-β receptor I kinase inhibitor. Next, the Boyden chamber assay was used to determine how TGF-β(2) (10 ng/ml) affect HCEC migration. Exposure to TGF-β(2) increased cell migration, and a synergistic effect was observed when FGF-2 was added. To determine whether the mitogen-activated protein kinase (MAPK) signaling pathway is involved in the migration of HCECs, western blot analysis and Bio-Plex(TM) suspension array were used to detect phosphorylation of Erk1/2, p38, and JNK in HCECs stimulated by TGF-β(2) and/or FGF-2. The effect of the p38 MAPK inhibitor, SB239063 (10 μM), on TGF-β(2) and/or FGF-2 induced cellular migration was determined by the Boyden chamber assay. Both TGF-β(2) and FGF-2 induced p38 phosphorylation, and a synergistic effect was observed with exposure to both growth factors. SB 239063 inhibited TGF-β(2) and FGF-2-induced migration of HCECs. These results indicate that TGF-β(2) reduces proliferation but stimulates migration of cultured HCECs. In addition, TGF-β(2) and FGF-2 may have synergistic effects on the migration of HCECs mediated by p38 MAPK phosphorylation.
    Experimental Eye Research 12/2012; 108C. DOI:10.1016/j.exer.2012.11.018 · 2.71 Impact Factor
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    ABSTRACT: Purpose: To evaluate the disinfectant effect of methylene blue (MB)-mediated photodynamic therapy (PDT) on a pathogenic strain of Acanthamoeba. Methods: Acanthamoeba castellanii (ATCC 50370) used in this study were treated under one of four experimental conditions: light irradiation and incubation in MB (L+M+), light irradiation and incubation in physiologic solution (L+M-), incubation in MB only (L-M+), and incubation in physiologic solution (L-M-). M+ trophozoites were incubated in either 0.25 or 0.5 mM MB for 10 minutes. L+ organisms were irradiated for 30 minutes following incubation in solution. A halogen lamp (660 ± 10 nm) with a maximum output of 6 mW/cm(2) was used as the PDT light source. After treatment, antiacanthamoeba activity was evaluated by checking the respiratory activity of the amoeba with 5-cyano-2,3-tetrazolium chloride (CTC) staining. We also determined whether the effect of PDT with MB had been retained or augmented when it was performed in combination with conventional antiamoebic agents. Results: MB-PDT suppressed the respiratory activity of trophozoites in an MB-concentration-dependent manner at total light doses of 10.8 J/cm(2). The respiratory activity of each group as a percentage of that of L-M- is as follows: L+M+ 11.6% (0.5 mM), 60.9% (0.25 mM); L-M+ 116.5% (0.5 mM), 105.5% (0.25 mM); L+M- 107.6%; and L-M- 106.3%. (L+M+ versus L-M- P < 0.05). MB-PDT had a synergistic effect when used in combination with polyhexamethylene biguanide (PHMB) or amphotericin B, but not with voriconazole. Conclusions: MB-PDT is effective against Acanthamoeba in vitro and has synergistic effects with PHMB and amphotericin B.
    Investigative ophthalmology & visual science 08/2012; 53(10):6305-13. DOI:10.1167/iovs.12-9828 · 3.40 Impact Factor
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    ABSTRACT: To assess a newly developed eyelid pressure measurement system called a blepharo-tensiometer that uses a tactile pressure sensor. The tactile sensor was 10 mm in diameter and approximately 0.4 mm thick. The sensor was covered with silicon rubber and was placed between a soft contact lens on the cornea and the inner surface of the eyelid. Under these conditions, the sensor measured the pressure of the eyelid on the ocular surface. The pressure of the upper and lower eyelids were measured separately while the eyelids were closed for at least 5 seconds in 34 eyes of 34 normal volunteers. To determine the reliability of the blepharo-tensiometer, the pressures of the upper and lower eyelids were measured on 3 separate days in both eyes of 12 normal volunteers. The correlation between the age and the eyelid pressure was calculated. The intraclass correlation coefficients for the 3 measurements ranged from 0.675 to 0.911 for the upper eyelid and 0.663 to 0.925 for the lower eyelid. The mean eyelid pressure was 16.95±6.08 mm Hg for the upper lid and 16.11±7.27 mm Hg for the lower lid. The eyelid pressure decreased with increasing age, and both the upper and lower eyelid pressures were significantly and negatively correlated with age (upper eyelid pressures, P<0.0001; lower eyelid pressures, P=0.000432). No complication was detected after the measurements in all of the subjects. Our blepahro-tensiometer can obtain repeatable measurements of the eyelid pressure and can be used to evaluate the pressure of the eyelids on the ocular surface in normal and diseased eyes.
    Eye & contact lens 08/2012; 38(5):326-30. DOI:10.1097/ICL.0b013e318268610a · 1.47 Impact Factor
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    ABSTRACT: To determine whether polymorphisms in the Toll-like receptor 4 (TLR4) gene are associated with primary open-angle glaucoma (POAG), normal-tension glaucoma (NTG), and exfoliation glaucoma (XFG) in Japanese individuals. Genetic association study. Setting: Multicenter study. Study population: One hundred eighty-four unrelated Japanese patients with POAG, 365 unrelated patients with NTG, and 109 unrelated patients with XFG from 5 hospitals. Procedures: Genomic DNA was extracted from leukocytes of the peripheral blood, and 8 polymorphisms in the TLR4 genes were amplified by polymerase chain reaction (PCR) and directly sequenced. Allele and genotype frequencies and the inferred haplotypes were estimated. Main outcome measures: Differences in allele and genotype frequencies and haplotypes between subjects with POAG, NTG, and XFG. The allele frequency of rs2149356 of the TLR4 gene in the POAG, NTG, and XFG groups was the most significantly different from that of the control group (minor allele frequency 0.446, 0.395, 0.404, vs 0.308; P = .000058, P = .0030, and P = .015). The allele frequencies of the 5 TLR4 SNPs were higher in all of the glaucoma groups than that in the control group. The statistics of genotypes of TLR4 were approximately the same for all allele frequencies. The haplotypic frequencies with Tag SNPs studied earlier showed that only POAG was statistically significant. Other haplotypes, such as rs10759930, rs1927914, rs1927911, and rs2149356, had higher statistical significance (overall P = .00078 in POAG, overall P = .018 in NTG, and overall P = .014 in XFG). This study demonstrated that TLR4 polymorphisms are associated with NTG in the Japanese, and they also play a role in the pathogenesis of POAG and XFG.
    American Journal of Ophthalmology 07/2012; 154(5):825-832.e1. DOI:10.1016/j.ajo.2012.03.050 · 3.87 Impact Factor

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  • 2006–2014
    • Ehime University
      • Department of Ophthalmology
      Matuyama, Ehime, Japan
  • 2009
    • Japanese Red Cross
      Edo, Tōkyō, Japan
  • 1997–2004
    • University of Cincinnati
      • Department of Ophthalmology
      Cincinnati, OH, United States
  • 2003
    • Wakayama Medical University
      • Department of Ophthalmology
      Wakayama, Wakayama, Japan