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Biomedical subjects

Yasuo Tano

Publications and source records attributed to Yasuo Tano.

At least 37 records · Page 2Linked to original sources

Paradoxical increase of visual impairment with punctal occlusion in a patient with mild dry eye.

We report a paradoxical increase in visual impairment after punctal plug placement despite improvement in corneal epithelial damage caused by mild dry eye. Ocular higher-order aberrations (HOAs) were measured sequentially with a wavefront sensor before and after punctal plug insertion in a patient with mild dry eye. Although postblink HOAs tend to increase in normal subjects or patients with dry eye, after treatment there was a maximum spike in HOAs for a few seconds that decreased gradually. Excessive retention of tear film by punctal occlusion may cause inferosuperior asymmetrical distribution in precorneal tear-film thickness, possibly leading to slower wavefront on the inferior cornea and increased coma-like aberrations. Increased visual impairment might occur paradoxically despite improvement in the corneal epithelial damage caused by dry eye.

Adult↗

Protective effect of donepezil on retinal ganglion cells in vitro and in vivo.

The neuroprotective effect of donepezil, an acetylcholinesterase inhibitor on retinal ganglion cells (RGCs), was examined. The survival of purified RGCs after exposure to glutamate with or without donepezil (10(- 7) M to 10(- 5) M) was measured after 3 days in culture. In vivo, the neuroprotective effect of donepezil was examined by quantifying the number of viable RGCs 7 days after axotomy in adult rats. In vitro, donepezil dose-dependently reduced RGC death caused by glutamate toxicity. Oral administration of donepezil (10 mg kg(-1) day(-1)) significantly reduced RGC death after axotomy. In conclusion, donepezil exerts a protective effect on RGCs both in vitro and in vivo.

Administration, Oral↗

Corneal epithelial stem cell delivery using cell sheet engineering: not lost in transplantation.

Cell-based therapies have now generated significant interest as novel drug delivery systems, with various adult cell types used in treating a wide range of diseases. To overcome the limits that restrict treatments for corneal surface dysfunction, corneal epithelial stem cells expanded ex vivo have been applied as an alternative approach. While previous studies used various carrier substrates, we present a novel method using cell sheet engineering with temperature-responsive culture dishes to create carrier-free corneal epithelial stem cell sheets that can be transplanted without sutures. Results from clinical trials reveal successful transplantation with the recovery of lost visual acuity in all cases. Cell sheet engineering, therefore, presents a novel method for the delivery of corneal epithelial stem cells, and can also be applied for other approaches of cellular therapeutics.

Corneal Transplantation↗

Wavefront analysis of eyes with cataracts in patients with monocular triplopia.

PURPOSE: To determine whether wavefront analysis using a Hartmann-Shack (H-S) aberrometer can reveal the cause of monocular triplopia in eyes with mild cataracts. METHODS: Six patients (nine eyes; age range 38-58 years; average 49.8 +/- 6.9 years) who complained of monocular triplopia at the Osaka University Hospital between January and December 2003 were examined. Wavefront analyses of ocular and corneal aberrations of the central 4 mm diameter were performed using a H-S aberrometer equipped with a Placido ring videokeratoscope. The ocular and corneal higher-order wavefronts were fitted with a fourth-order Zernike expansion. RESULTS: All nine eyes showed mild nuclear cataract and had a mean spherical refractive error of -10.3 +/- 3.5 D. The visual acuity was > or = 20/40 except in one eye with glaucoma. For the Zernike polynomials, the trefoil aberration (C3-3) and the spherical aberration (C40) were significantly higher than those of age-matched normal controls (p < 0.001). The simulated retinal image of a Landolt C showed that the combination of trefoil aberration and the spherical aberration can cause an image with a triple configuration. CONCLUSIONS: Monocular triplopia was reported by middle-aged patients with mild nuclear cataract and high myopia. Wavefront analyses suggested that the triple configuration was caused by the combined increase of the trefoil and spherical aberration in lenses with mild nuclear cataracts.

Adult↗

Transplantation of tissue-engineered epithelial cell sheets after excimer laser photoablation reduces postoperative corneal haze.

PURPOSE: To apply tissue-engineered cell sheet transplantation after excimer laser keratectomy as a novel approach for the reduction of postoperative corneal haze. METHODS: Limbal biopsy specimens were obtained, and epithelial cells were cultured on temperature-responsive culture inserts without the use of feeder cells. Laser keratectomy (7.0-mm ablation zone and 160-microm depth) was performed in the contralateral eye, and autologous epithelial cell sheets were transplanted to the ablated corneal stroma. Transplant and control group eyes were assessed by slit lamp biomicroscopy, and corneal haze was scored in a masked fashion, according to the Fantes grading scale. For further examination histologic and immunohistochemical analyses were performed. RESULTS: Tissue-engineered cell sheets produced stable attachment to the laser-ablated sites, resulting in epithelialization, 5 minutes after transplantation. Conversely, control corneas required 3 to 5 days for complete re-epithelialization. At both 1 and 2 months after surgery, corneal haze was significantly inhibited in the transplant group. Histologic analyses showed that the number of keratocytes undergoing apoptosis was decreased in the transplant group at 3 days after surgery. Similarly, the expression of both collagen III and alpha-smooth muscle actin, which may enhance corneal haze, were diminished in the transplant group at 2 months. CONCLUSIONS: The transplantation of tissue-engineered epithelial cell sheets can successfully prevent the development of corneal haze after excimer laser keratectomy.

Actins↗

Serial measurements of higher-order aberrations after blinking in normal subjects.

PURPOSE: To investigate sequential changes in the optical quality of normal eyes associated with blinking. METHODS: Ocular higher-order aberrations (HOAs) were measured sequentially by using a wavefront sensor for 30 seconds in 20 eyes of 20 normal subjects. During the measurement, subjects were forced to blink every 10 seconds. The obtained aberration data were analyzed in the central 4-mm diameter for coma-like, spherical-like, and total HOAs up to the sixth-order Zernike polynomials. RESULTS: The serial changes in the HOAs with blinking were classified into four groups by pattern: stable (25%), small-fluctuation (45%), sawtooth (20%), and others (10%). In the subjects with the sawtooth pattern, the total HOAs increased significantly (P < 0.001, one-way repeated-measures ANOVA) with time between blinks. Increased total HOAs and coma-like aberrations in the subjects with the sawtooth pattern suggested that the inferosuperior asymmetric change in tear film thickness is responsible. CONCLUSIONS: Dynamic changes in HOAs after blinking showed variations even in clinically normal subjects. Serial measurements of HOAs may be useful in evaluating the dynamic changes in tear film and the effects on the quality of vision after blinking.

Adult↗

[Sub-Tenon's injection of triamcinolone acetonide for choroidal neovascularization].

PURPOSE: To evaluate the efficacy of sub-Tenon's injection of triamcinolone acetonide for age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV). METHOD: Sub-Tenon's injection of triamcinolone acetonide was performed in patients (27 eyes) with the subfoveal choroidal neovascularization of AMD and in patients (10 eyes) with PCV, and its efficacy was evaluated. The visual acuity and the exudation from lesions evaluated by fluorescein angiography or optical coherence tomography (OCT) at three months or six months after injection were compared to the baseline values. RESULTS: Visual acuity was maintained in 23 of the 27 eyes (85.0%) with AMD and in 8 of the 10 eyes (80.0%) with PCV at three months after injection. Six months after infusion, the rate declined to 76.0% and 66.7%, respectively. Exudation from lesions had decreased in 10 eyes of 25 eyes(40.0%) with AMD and in 2 eyes of 9 eyes (22.2%) with PCV by six months after injection. Complete suppression of the lesions was observed in seven eyes (18.9%) with relatively small lesions. CONCLUSION: Sub-Tenon's injection of triamcinolone acetonide is useful as a treatment modality for AMD and PCV.

Aged↗

Surgical outcomes of 25-gauge transconjunctival vitrectomy combined with cataract surgery for vitreoretinal diseases.

INTRODUCTION: To report surgical outcomes of 25-gauge transconjunctival vitrectomy combined with cataract surgery for the management of a variety of vitreoretinal diseases. MATERIALS AND METHODS: A retrospective, interventional case study was conducted. Chart review of a consecutive series of 150 eyes of 144 patients who underwent 25-gauge vitrectomy combined with phacoemulsification and intraocular lens implantation for epiretinal membrane (n = 62), refractory macular oedema associated with retinal vascular disorders (n = 29), idiopathic macular hole (n = 21), non-clearing vitreous haemorrhage (n = 18), rhegmatogenous retinal detachment (n = 11), tractional retinal detachment associated with proliferative diabetic retinopathy (n = 7), and subretinal haemorrhage (n = 2). Main outcome measures included preand postoperative visual acuity, operating time, intraocular pressure, intra- and postoperative complications. RESULTS: The mean follow-up period was 9.7 months (range, 6 to 26). The mean overall visual acuity improved from 20/100 preoperatively to 20/38 at final visit (P <0.001). Statistically significant improvement of visual acuity was also observed in each subgroup. Operative time was shortened in macular surgery. No intraoperative complications were noted attributable to small-gauge instruments and no cases required conversion to 20-gauge standard instrumentation. However, 12 eyes (8%) required suture placement to at least one sclerotomy site. Postoperative intraocular pressure remained stable in most cases except 18 eyes (13%) with transient hypotony during the first week after surgery. One case of retinal detachment but no case of endophthalmitis was observed throughout the follow-up period. CONCLUSIONS: 25-gauge vitrectomy combined with cataract surgery is a safe and effective system for the management of a variety of vitreoretinal diseases, especially cases requiring minimal intraocular manipulation. Further study is recommended to evaluate potential postoperative complications.

Aged↗

[Clinical study of anterior ocular segment topography in angle-closure glaucoma using the three-dimensional anterior segment analyzer Pentacam].

PURPOSE: The anterior ocular segment topography in angle-closure glaucoma was studied using the non-contact three-dimensional anterior segment analyzer Pentacam. SUBJECTS AND METHODS: The central anterior chamber (AC) depth, mid-peripheral AC depth, peripheral AC depth, AC volume, central AC depth/ocular axial length ratio and angles of a narrow angle group (47 eyes of 29 patients), a post laser iridotomy (LI) group (43 eyes of 30 patients), and an open-angle glaucoma (OAG) group (37 eyes of 21 patients) were measured and compared. In addition, changes in the parameters before and after LI (13 eyes of 9 patients) were investigated. RESULTS: The AC volume for the narrow angle group (74.5 +/- 21.1 microl) was significantly smaller than for the other groups (post LI group: 96.4 +/- 21.4 microl; OAG group: 144.2 +/- 31.6 microl, p<0.001). The most significant association was detected between AC volume and the peripheral AC depth. Only two parameters, AC volume and peripheral AC depth, increased significantly after LI (p< 0.001). DISCUSSION: Measurement of the AC volume and the peripheral AC depth using Pentacam is useful for evaluating the anterior ocular segment topography in narrow angle eyes.

Aged↗

Functional human corneal endothelial cell sheets harvested from temperature-responsive culture surfaces.

This study reports a new method for fabricating bioengineered human corneal endothelial cell sheets suitable for ocular surgery and repair. We have initially cultured human corneal endothelial cells on type IV collagen-coated dishes and, after several passages, expanded cells were then seeded onto novel temperature-responsive culture dishes. Four weeks after reaching confluence, these cultured endothelial cells were harvested as intact monolayer cell sheets by simple temperature reduction without enzymatic treatment. Scanning electron microscopy indicated that these cells were primarily hexagonal with numerous microvilli and cilia, similar to the native corneal endothelium. The Na+, K+-ATPase pump sites were located at the cell borders as in vivo. Moreover, cell densities and numbers of pump sites were identical to those of in vivo human corneal endothelium under optimized conditions. A 3H-ouabain binding analysis demonstrated a linear proportionality for cell pump density between confluent cell densities of 575 cells/mm2 and 3070 cells/mm2. We also confirmed Na+, K+-ATPase activity in the sheets in vitro. Xenograft transplantation results showed that the fabricated sheets retain their function of maintaining proper stromal hydration in vivo. We have established a regimen to culture and proliferate human corneal endothelial cells and fabricate endothelial sheets ex vivo morphologically and functionally similar to the native corneal endothelium. Our results support the value of harvested cell sheets for clinical applications in ocular reconstructive surgery in patients with ocular endothelial decompensation.

Cell Culture Techniques↗

p57Kip2 is expressed in quiescent mouse bone marrow side population cells.

Hematopoietic stem cells can be accurately identified by the side population (SP) phenotype. It has been previously shown that hematopoietic stem cells are cell cycle arrested, but the mechanisms involved are currently poorly understood. In the present study, results from quantitative real-time RT-PCR show that while SP cells have increased expression of various cyclins and cyclin-dependent kinases, the increased expression of cyclin-dependent kinase inhibitors, in particular p57(Kip2), is responsible for the observed cell cycle arrest. In addition, gene expression analysis of c-kit(+/)/Sca-1(+)/Lineage- SP (KSL-SP) cells demonstrates that only p57(Kip2) shows both higher expression compared to both SP and non-SP cells. Furthermore, immunostaining also demonstrates significantly higher protein expression in KSL-SP cells. These results demonstrate that the maintenance of bone marrow SP cells in G0/G1 may be carefully controlled by p57(Kip2).

Animals↗

Mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K) pathways differently regulate retinal pigment epithelial cell-mediated collagen gel contraction.

Retinal pigment epithelial (RPE) cell-mediated extracellular matrix contraction is believed to contribute to developing proliferative vitreoretinopathy. It has been shown that platelet-derived growth factor (PDGF) and its intracellular signaling pathway, including mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K), are mainly involved in this process. The aim of this study is to investigate how these downstream signaling pathways are related to RPE-mediated collagen gel contraction. We performed the gel contraction assay to evaluate the effect of PDGF in cultured ARPE-19 cells under the presence or absence of PD98059, MAPK inhibitor or wortmannin, PI3K inhibitor. Experiments treated with neutralizing antibody for various subtypes of integrin were also performed and the effect on PDGF-induced gel contraction was investigated. Expression changes of integrin alpha1, alpha2 and beta1 after PDGF stimulation was evaluated using quantitative real-time PCR and flow cytometry. The results showed that PDGF up-regulated ARPE-19 cell-mediated gel contractile activity. PDGF-induced collagen gel contraction was attenuated under presence of PD98059, wortmannin, or neutralizing antibody for integrin alpha1, alpha2, or beta1, all of which are critical subset for binding with type I collagen. The expression of integrin alpha1 and alpha2 was increased after PDGF stimulation in both real-time PCR and flow cytometry, however beta1 expression was not increased. PD98059 significantly attenuated integrin alpha1 and alpha2 expressions. However, wortmannin did not have the same effect. In conclusion, PDGF promotes ARPE-19 cell-mediated gel contraction via both MAPK and PI3K. This was probably due to an increased expression of integrin alpha1 and alpha2, which is mediated by MAPK, but not by PI3K. PI3K may regulate collagen gel contraction by another mechanism other than the up-regulation of integrin expression.

Androstadienes↗

Rat limbal epithelial side population cells exhibit a distinct expression of stem cell markers that are lacking in side population cells from the central cornea.

The side population (SP) phenotype is shared by stem cells in various tissues and species. Here we demonstrate SP cells with Hoechst dye efflux were surprisingly collected from the epithelia of both the rat limbus and central cornea, unlike in human and rabbit eyes. Our results show that rat limbal SP cells have a significantly higher expression of the stem cell markers ABCG2, nestin, and notch 1, compared to central corneal SP cells. Immunohistochemistry also revealed that ABCG2 and the epithelial stem/progenitor cell marker p63 were expressed only in basal limbal epithelial cells. These results demonstrate that ABCG2 expression is closely linked to the stem cell phenotype of SP cells.

ATP Binding Cassette Transporter, Subfamily G, Mem↗

Limbal epithelial side-population cells have stem cell-like properties, including quiescent state.

Corneal epithelial (CE) stem cells are believed to reside in the basal layer of the limbal epithelium but remain poorly understood due to the lack of an accepted in vivo reconstitution assay as well as definitive markers for epithelial stem cells. It has been reported that side-population (SP) cells with the ability to efflux the DNA-binding dye Hoechst 33342 have stem cell-like properties and that the SP phenotype accurately represents a quiescent and immature stem cell population in the adult bone marrow. In the present study, we investigated whether SP cells isolated from the limbal epithelium have stem cell-like properties. SP cells, separated by fluorescence-activated cell sorting, comprise approximately 0.4% of all limbal epithelial cells and have markedly higher expression of the stem cell markers ABCG2, Bmi-1, and nestin but no expression of markers for differentiated CE cells compared with non-SP cells. Cell-cycle and telomerase activity analyses revealed that SP cells are growth arrested and reside in the quiescent state. Moreover, limbal epithelial SP cells did not demonstrate proliferative capabilities under typical in vitro epithelial cell culture conditions using 3T3 feeder layers. These findings present the possibility that quiescent limbal epithelial SP cells may represent an extremely immature stem cell population compared with currently defined epithelial stem or progenitor cells.

Animals↗

Structural characterization of bioengineered human corneal endothelial cell sheets fabricated on temperature-responsive culture dishes.

For the purpose of corneal regenerative medicine, we fabricated human corneal endothelial cell sheets on temperature-responsive dishes, which could be non-invasively harvested as intact, transplantable sheets by simply reducing the culture temperature. Cells demonstrated hexagonal cell shape with numerous microvilli and cilia, and also exhibited abundant cytoplasmic organelles similar to these cells in vivo. Immunofluorescence for type IV collagen and fibronectin revealed that abundant extracellular matrix (ECM) was deposited on the basal surface throughout culture, and the deposited ECM was harvested along with the cell sheets by reducing culture temperature to 20 degrees C. Faint ECM remnants were observed on the dish surfaces after cell sheet detachment. Immunofluorescence for ZO-1 showed that tight junctions were established between cells, and immunoblotting indicated that intact ZO-1 was maintained during cell sheet harvest, while conventional proteolytic cell harvest methods resulted in the degradation of ZO-1. These results suggest that these transplantable corneal endothelial cell sheets can be applied to treat patients with damaged corneas.

Cell Culture Techniques↗

Electrophysiological and histological studies of chronically implanted intrapapillary microelectrodes in rabbit eyes.

PURPOSE: To determine the safety and efficacy of transsclerally placed intrapapillary wire microelectrodes implanted chronically into the optic nerve head of rabbit eyes. METHODS: Four platinum wire microelectrodes were passed through the sclera and implanted into the optic nerve head of five rabbit eyes for 4-6 months. Color fundus photography, fluorescein angiography, electroretinograms (ERGs), and visually evoked potentials (VEPs) were used to monitor the retina. Electrically evoked potentials (EEPs) were elicited by bipolar electrical stimulation of the optic nerve axons by different combinations of the four electrodes immediately after the implantation and at 1-month intervals thereafter. The effects of the chronic implantation of the electrodes on the morphology of the optic nerve were evaluated by histological and immunohistochemical examinations at 4 and 6 months after the implantation. RESULTS: All of the electrodes remained stable in the implanted sites throughout the post-implantation period, except for one electrode that had pulled out of the optic nerve head at 1 month after implantation. No intraocular infection, inflammation, or vitreoretinal proliferation was observed in any eye. EEPs could be elicited from each pair of electrodes at all testing times. The mean threshold currents (charge densities) to evoke EEPs increased from 19.3 +/- 9.2 microA (6.0 +/- 2.9 microC/cm2) on the implantation day to 78.8 +/- 31.9 microA (24.6 +/- 10.0 microC/cm2) at 1 month after implantation, but did not change significantly thereafter. The implicit time and amplitude of the a- and b-waves of the ERGs and of P1 of the VEPs did not change significantly throughout the post-implantation period. Histological evaluation of the optic nerve head revealed slight tissue encapsulations surrounding the electrode and increased expression of glial fibrillary acidic protein near the surface of the optic nerve. CONCLUSIONS: Implantation of transscleral intrapapillary microelectrodes appears to be safe and effective. These findings indicate that the implantation of microelectrodes in the optic nerve head should be considered for an optic nerve-based prosthesis.

Animals↗

A novel missense mutation in a Japanese patient with gelatinous droplike corneal dystrophy.

PURPOSE: To report a novel missense mutation in TACSTD2 gene, L186P, responsible for gelatinous droplike dystrophy (GDLD). DESIGN: Case report and experimental study. METHOD: A 10-year-old Japanese boy suffering from typical GDLD was studied. A 1.1-kb DNA fragment of the TACSTD2 gene was amplified and analyzed using a molecular biological method. cDNA from the patient's cornea was also analyzed to determine which allele was expressed in the patient's corneal epithelium. RESULTS: Sequence analysis revealed that the patient is a compound heterozygote for the Q118X mutation and the L186P, the first missense mutation found in Japanese GDLD. Polymerase chain reaction-restriction fragment length polymorphism analysis from cDNA of patient's cornea revealed that the L186P missense mutation allele is expressed in the patient's corneal epithelium. CONCLUSION: We describe a novel mutation in one case of Japanese GDLD. The results confirm that the missense mutation L186P in the TACSTD2 gene is also responsible for the GDLD phenotype.

Antigens, Neoplasm↗