Three-dimensional optical coherence tomographic findings of idiopathic multiple serous retinal pigment epithelial detachment.
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Publications and source records attributed to Yasuo Tano.
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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.
PURPOSE: To reveal the pathogenesis of myopic foveoschisis (MF). DESIGN: Clinicopathological report. METHODS: Internal limiting membranes (ILMs) were collected from ten patients with MF and five patients with idiopathic macular hole (IMH) as a control. Samples were subjected to transmission electron microscopic study. Characteristics of the ILM were compared between the two groups. RESULTS: Collagen fiber and cell debris were identified on the inner surface of ILM in seven eyes (70%) with MF, significantly more (P < .05) than found in IMH subjects (0%). More fibrous glial cells were likely to be found on the inner surface of ILM. No significant difference in fibroblast-like cell adhesion was observed. CONCLUSIONS: Collagen fiber and cellular component are suggested to play an important role in developing MF. ILM peeling may be essential for vitrectomy for MF.
PURPOSE: To develop new micro vertical scissors for the ablation of retinal angiomatous proliferation (RAP) to minimize possible retinal damage. DESIGN: Interventional case report. METHODS: New micro vertical scissors were developed and used for the ablation of RAP vessels. RESULTS: The length of the inner side of the tip is 180 microm, which is approximately one seventh that of regular vertical scissors. The scissors handled easily and the RAP vessels were ablated with less retinal damage compared with that associated with use of standard vertical scissors. Postoperatively, intraretinal hemorrhage was reduced and hyperfluorescence seen on indocyanine green angiography resolved. CONCLUSIONS: The new micro vertical scissors developed for the ablation of RAP vessels can be used safely and effectively during this surgical procedure.
PURPOSE: To report retinal microfold formation after vitrectomy for myopic foveoschisis (MF). DESIGN: Prospective observational study. METHODS: We observed 21 eyes of 17 patients who had undergone vitrectomy for MF with optical coherence tomography (OCT) in this institutional study. We also evaluated the three-dimensional retinal architecture using the OCT-ophthalmoscope in selected cases. Vitrectomy included core vitrectomy, vitreous cortex removal, internal limiting membrane (ILM) peeling with indocyanine green, and gas tamponade. RESULTS: Horizontal linear folds were commonly observed postoperatively. The folds, which were 1,000 to 2,000 microm superior, inferior, or both superior and inferior to the fovea, were detected only by OCT and not by conventional slit-lamp-based biomicroscopy. The microfolds were found in only five eyes (24%) 1 month postoperatively. The incidence increased over time, however, and a microfold was detected in nine eyes (43%) 3 months after surgery and in 13 (62%) 6 months after surgery. OCT-ophthalmoscope examination confirmed the location of the microfold coincided exactly with that of retinal arteriole. The presence of microfolds was not significantly related to the postoperative visual acuity. CONCLUSIONS: Retinal microfolds are common in eyes with MF after vitrectomy with ILM peeling, and they seem to be generated as the result of insufficient flexibility of the sclerotic retinal arteriole during axial length elongation in highly myopic eyes. This finding suggests that the inward tractional force on the retina along the arteriole may be closely related to the pathogenesis of vitreoretinal diseases specific to high myopia, including MF or paravascular microhole formation.
PURPOSE: To report a case of retinal angiomatous proliferation (RAP) treated by ablation of the feeding and draining vessels that recurred by 6 months postoperatively. DESIGN: Interventional case report. METHODS: Images from fluorescein angiography, indocyanine green (ICG) angiography, and optical coherence tomography (OCT) were examined before and after ablation. RESULTS: The hot spot on late-phase ICG, the leakage on fluorescein angiography, the pigment epithelial detachment, and macular edema on OCT decreased postoperatively. However, a large hyperfluorescent lesion was detected on ICG 6 months postoperatively with recurrent cystoid macular edema. CONCLUSIONS: A new RAP lesion can develop even after the original lesion seemed to resolve after ablation.
PURPOSE: Retinal microfolds attributable to retinal vessels were first observed after vitrectomy for myopic foveoschisis and were believed to indicate inward retinal traction, possibly leading to high myopia-specific retinal diseases. We report retinal microfolds in high myopia without vitrectomy. DESIGN: Observational case series. METHODS: This is an institutional study. Seven eyes of seven patients in which retinal microfolds were observed using optical coherence tomography (OCT) were included in the study. We used an OCT-ophthalmoscope to confirm the precise location of the microfolds. We also investigated the relationship between the presence of microfolds and subjective distortion detected by examination of the corresponding retinal area using the Amsler grid chart in three eyes. RESULTS: All microfolds detected by OCT coincided with the retinal vessels using the OCT-ophthalmoscope. The folds coincided only with the retinal arterioles in six eyes (86%) and with both arterioles and veins in one (14%). Subjective distortion was detected in the retinal area corresponding to the microfolds in two eyes (67%) but was not detected in one eye (33%). CONCLUSIONS: The incidence of retinal microfolds is 2.9%, and thus they are not uncommon in highly myopic eyes without vitrectomy. The coincident appearance of the folds and vessels suggests that inflexibility of the retinal vessels and retinal stretching attributable to ocular elongation may cause the microfolds. The presence of these microfolds indicates that inward retinal vascular traction could be common in highly myopic eyes.
PURPOSE: To report the long-term follow-up results of nonvitrectomizing vitreous surgery for idiopathic epiretinal membrane (ERM). DESIGN: Nonrandomized comparative case series. PARTICIPANTS: Thirty patients were followed up for at least 5 years after nonvitrectomizing vitreous surgery. INTERVENTION: Epiretinal membranes were peeled without infusion of balanced salt solution and removal of the vitreous. The data from the fellow eye was the control data. MAIN OUTCOME MEASURES: We examined the visual acuities (VAs), objective refractions, and slit-lamp and Scheimpflug photographs from the preoperative and the final examinations of both eyes. Quantitative assessment of the progression of nuclear sclerosis was performed by densitometry analysis using Scheimpflug photography. The recurrence rate of ERM was determined. RESULTS: The follow-up periods ranged from 60 to 102 months (mean+/-standard deviation, 72.2+/-11.0 months). The patient ages ranged from 52 to 76 years (68.8+/-6.3 years). The final VA improved or stabilized within 2 lines in 29 of 30 eyes (96.7%). No unilateral progression of nuclear sclerosis occurred in any cases. The mean preoperative and postoperative refractions without additional surgery were -0.4+/-2.9 diopters (D) and -0.2+/-3.0 D in the operated eyes, respectively, and -0.2+/-2.5 D and 0.1+/-2.4 D, respectively, in the unoperated fellow eyes. The mean differences in the refractive error between both eyes (operated eye data minus fellow eye data) were -0.2+/-0.7 D before surgery and -0.3+/-0.8 D after surgery (P = 0.319, paired t test). The mean preoperative and postoperative nuclear densities in 16 patients were 69+/-14 nuclear density units (NDUs) and 76+/-12 NDUs in the operated eyes and 71+/-14 NDUs and 78+/-14 NDUs in the fellow eyes, respectively. The mean preoperative and postoperative differences in nuclear densities in both eyes were -2+/-2 NDUs and -2+/-5 NDUs, respectively (P = 0.836, paired t test). The ERM recurred in 10 eyes (33%), and 3 eyes underwent conventional vitrectomy combined with cataract surgery. CONCLUSIONS: Unilateral nuclear sclerosis did not progress for at least 5 years after nonvitrectomizing vitreous surgery. The recurrence rate of ERM appeared to be higher than that after conventional vitreous surgery.
PURPOSE: To investigate whether triamcinolone acetonide (TA) affects the expression of vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) in retinal pigment epithelial (RPE) cells exposed to oxidative stress. METHODS: TA (10 nM, 1 microM, or 100 microM) was added to ARPE19 cells exposed to oxidative stress induced by hypoxia-reoxygenation and paraquat. Cellular expression of VEGF, CTGF, and an inducer of both growth factors, transforming growth factor (TGF)-beta was investigated with real-time reverse transcription-polymerase chain reaction and Western blot analysis. Tube-forming assays were conducted with human umbilical vein endothelial cells (HUVECs) in conditioned medium from RPE cells exposed to oxidative stress, with or without TA treatment. RESULTS: Oxidative stress induced mRNA expression of VEGF, CTGF, and TGF-beta by RPE cells. TA reduced upregulation of VEGF and TGF-beta in a concentration-dependent manner. In contrast, upregulation of CTGF by oxidative stress was accelerated by TA concentrations of 10 nM and 1 muM. Tube formation by HUVECs was strongly inhibited by exposure to conditioned medium from oxidative stress-stimulated ARPE19 cells treated with 1 muM TA compared with cells not treated with TA. CONCLUSIONS: TA reduced VEGF expression and induced CTGF expression in ARPE19 cells exposed to oxidative stress, and conditioned medium from these cells inhibited tube formation by HUVECs. Because VEGF is a major cytokine involved in angiogenesis, and CTGF is a main cytokine related to fibrosis, these results suggest that changes in their expression may be important mechanisms underlying the decreased choroidal neovascularization and fibrosis after administration of TA.
PURPOSE: Autologous stem cell transplantation for total limbal stem cell deficiency is immunologically preferable, to avoid allograft rejection. This study was undertaken to investigate the possibility of a novel tissue engineering approach for ocular surface reconstruction, using autologous oral mucosal epithelial stem cells expanded ex vivo on temperature-responsive cell culture surfaces. METHODS: Rabbit oral mucosal epithelial cells cultured on temperature-responsive culture surfaces with mitomycin-C-treated 3T3 feeder cells for 2 weeks produced confluent epithelial cell sheets. Putative progenitor cell populations were estimated by colony-forming assays. Autologous transplantation of these cell sheets to surgically manipulated eyes was performed, and ocular surface reconstruction and cell phenotypic modulation were examined. RESULTS: All cultured oral epithelial cells were nonenzymatically harvested as transplantable intact cell sheets by reducing culture temperature to 20 degrees C. Oral epithelial cells were stratified in three to five cell layers more similar to corneal epithelium than to oral mucosal epithelium. Colony-forming assays and immunofluorescence for p63, beta1-integrin, and connexin 43 indicated retention of viable stem and/or progenitor cell populations in cell sheets. Autologous transplantation to rabbit corneal surfaces successfully reconstructed the corneal surface, with restoration of transparency. Four weeks after transplantation, epithelial stratification was similar to that in the corneal epithelium, although the keratin expression profile retained characteristics of the oral mucosal epithelium. CONCLUSIONS: Cell sheet harvest technology enables fabrication of viable, transplantable, tissue-engineered epithelial cell sheets that retain putative progenitor cells from autologous oral mucosal epithelial cells. Promising clinical capabilities for autologous tissue-engineered epithelial cell sheets for ocular surface reconstruction are indicated.
PURPOSE: To investigate the effect of transcorneal electrical stimulation (TES) on the survival of axotomized RGCs and the mechanism underlying the TES-induced neuroprotection in vivo. METHODS: Adult male Wistar rats received TES after optic nerve (ON) transection. Seven days after the ON transection, the density of the surviving RGCs was determined, to evaluate the neuroprotective effect of TES. The levels of the mRNA and protein of insulin-like growth factor (IGF)-1 in the retina after TES were determined by RT-PCR and Northern and Western blot analyses. The localization of IGF-1 protein in the retina was examined by immunohistochemistry. RESULTS: TES after ON transection increased the survival of axotomized RGCs in vivo, and the degree of rescue depended on the strength of the electric charge. RT-PCR and Northern and Western blot analyses revealed a gradual upregulation of intrinsic IGF-1 in the retina after TES. Immunohistochemical analysis showed that IGF-1 immunoreactivity was localized initially in the endfeet of Muller cells and then diffused into the inner retina. CONCLUSIONS: TES can rescue the axotomized RGCs by increasing the level of IGF-1 production by Muller cells. These findings provide a new therapeutic approach to prevent or delay the degeneration of retinal neurons without the administration of exogenous neurotrophic factors.
BACKGROUND: A new method of stimulating the retina electrically, called suprachoroidal transretinal stimulation (STS), was shown to be effective in eliciting electrically evoked cortical potentials (EEPs) in Royal College of Surgeons (RCS) rats. Before extending this technique to patients, it is important to determine its safety and feasibility in eliciting EEPs from medium-size animal (rabbits). The purpose of this study was to determine the safety and efficacy of the surgical procedures used to implant an multichannel electrode array into a scleral pocket, and to determine whether the implanted electrodes can stimulate the retina effectively. METHODS: These acute experiments were conducted on six rabbits. An array of eight gold microelectrodes, embedded in polyimide, was implanted into a scleral pocket over the visual streak area. The size of the microarray was 2 x 4 x 0.180 mm. The reference electrode was implanted into the vitreous. The electrode array and reference electrodes were connected to a stimulator to deliver monophasic current pulses. Cortical responses were recorded with a stainless steel electrode implanted into each rabbit's skull over the visual cortex. After the experiment, the eyes and electrodes were examined histologically. RESULTS: The surgical procedures for electrode implantation were accomplished without serious complications. EEPs were recorded after monophasic electrical pulse stimulation from each electrode. The mean threshold for EEPs was 55.0+/-10.0 microA with a 0.5-ms duration inward current pulse. The charge delivered at threshold was about 27.5 nC, and the charge density was about 56.0 microC/cm2. Histopathological examination of the retinal tissue around the area of stimulation did not show damage at the light microscope level with the electrical parameters used. CONCLUSIONS: Our technique for STS with an intrascleral microelectrode array is safe in rabbit eyes, and EEPs were elicited by current densities that did not induce tissue damage. These results suggest that STS via intrascleral multichannel electrodes is a feasible method for stimulating the retina.
PURPOSE: To determine whether wire microelectrodes implanted in the optic disc can be used to elicit cortical potentials. METHODS: Two or four platinum wire electrodes of two types, viz., the cut-end type and the exposed-tip type, were inserted through the vitreous and fixed in the optic disc of 16 rabbit eyes. Electrically evoked potentials (EEPs) were recorded after bipolar electrical stimulation with the two wire electrodes and by different combinations of the four-electrode system. The optic discs were examined histologically after the experiment. RESULTS: The wire electrodes were successfully implanted and fixed into different positions of the optic disc without serious complications in all 16 eyes. EEPs could be elicited after bipolar electrical stimulation of the optic nerve using either the two-electrode system or different pairs of the four-electrode system. Threshold charge densities to elicit EEPs were 0.32-0.64 mC/cm(2) in eyes using the cut-end type of electrodes and 0.93-6.21 muC/cm(2) in eyes using the exposed-tip type. The amplitude of the EEPs increased with increasing electrical stimulus intensities. Histological evaluation revealed limited damage to the neural tissue adjacent to the electrode track. CONCLUSIONS: The visual cortex can be activated by direct microelectrical stimulation of the optic nerve. The acute implantation of the wire microelectrodes into the optic disc by a transvitreal approach is feasible and results in only limited damage to the optic nerve.
BACKGROUND: Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe loss of vision as a result of complete loss of corneal epithelial stem cells. Transplantation of autologous corneal stem-cell sources is an alternative to allograft transplantation and does not require immunosuppression, but it is not possible in many cases in which bilateral disease produces total corneal stem-cell deficiency in both eyes. We studied the use of autologous oral mucosal epithelial cells as a source of cells for the reconstruction of the corneal surface. METHODS: We harvested 3-by-3-mm specimens of oral mucosal tissue from four patients with bilateral total corneal stem-cell deficiencies. Tissue-engineered epithelial-cell sheets were fabricated ex vivo by culturing harvested cells for two weeks on temperature-responsive cell-culture surfaces with 3T3 feeder cells that had been treated with mitomycin C. After conjunctival fibrovascular tissue had been surgically removed from the ocular surface, sheets of cultured autologous cells that had been harvested with a simple reduced-temperature treatment were transplanted directly to the denuded corneal surfaces (one eye of each patient) without sutures. RESULTS: Complete reepithelialization of the corneal surfaces occurred within one week in all four treated eyes. Corneal transparency was restored and postoperative visual acuity improved remarkably in all four eyes. During a mean follow-up period of 14 months, all corneal surfaces remained transparent. There were no complications. CONCLUSIONS: Sutureless transplantation of carrier-free cell sheets composed of autologous oral mucosal epithelial cells may be used to reconstruct corneal surfaces and can restore vision in patients with bilateral severe disorders of the ocular surface.
Tenomodulin (TeM) is a type II transmembrane glycoprotein that contains a C-terminal domain with homology to the mature, secreted form of chondromodulin-I (ChM-I), a cartilage-derived angiogenesis inhibitor. TeM transcripts have been found in hypovascular tissues such as tendons and ligaments but the biological activity of TeM has not yet been fully explored. Using an adenovirus expression system, we utilized the forced expression and subsequent secretion of the human TeM C-terminal 116 amino acids (Ad-shTeM) in human umbilical vein endothelial cells (HUVECs) to assess the anti-angiogenic properties of TeM. The C-terminal 120 amino acids of the human ChM-I precursor (Ad-shChM-I) was similarly expressed in HUVECs as a comparison study. Transduction of both Ad-shTeM and Ad-shChM-I resulted in significant impairment of the tube-forming activity of HUVECs, when cultured in Matrigel. Similarly, conditioned medium from COS7 cells, transfected with plasmid DNA encoding shTeM or shChM-I, inhibited tube formation of HUVECs when compared to medium derived from either COS7 cells transfected with control vector or from non-transfected cells. Upon infection of HUVECs with Ad-shTeM or Ad-shChM-I, DNA synthesis stimulated by vascular endothelial growth factor (VEGF) was reduced to 40-50% of normal levels. Additionally, in a modified Boyden chamber assay, migration of HUVECs in response to VEGF was significantly affected following transduction of either Ad-shTeM or Ad-shChM-I and these transduced HUVECs were found to spread well on type I collagen or fibronectin, but not on vitronectin. Furthermore, the transduction of either Ad-shTeM or Ad-shChM-I in human melanoma cells resulted in suppression of tumor growth in association with decreased vessel density in vivo. Hence, we have demonstrated that, similarly to ChM-1, the C-terminal domain of TeM exhibits both anti-angiogenic and anti-tumor activities when expressed in a secreted form.
Many types of organ-specific stem cells have been recently shown to exhibit a side population (SP) phenotype based on their ability to efflux Hoechst 33342 dye. Because stem cells from corneal epithelium reside in the basal layer of the limbal epithelium, the purpose of this study was to examine whether the limbal epithelium contains SP cells. The ATP-binding cassette transporter Bcrp1/ABCG2 is reported to contribute to the SP phenotype in cells from several diverse sources. Here we show data from fluorescence-activated cell sorting and real-time quantitative RT-PCR analysis showing that harvested limbal epithelial cells contain SP cells expressing ABCG2. Immunofluorescence revealed that a portion of limbal epithelial basal cells expressed ABCG2. Data indicate that ABCG2 positive limbal epithelial cells are putative corneal epithelial stem cells.
BACKGROUND: Limbal stem-cell deficiency by ocular trauma or diseases causes corneal opacification and visual loss. Recent attempts have been made to fabricate corneal epithelial graft constructs, but the technology is still evolving. We have developed a novel cell-sheet manipulation technology using temperature-responsive culture surfaces to generate functional, cultivated corneal epithelial cell sheet grafts. METHODS: Human or rabbit limbal stem cells were cocultured with mitomycin C-treated 3T3 feeder layers on temperature-responsive culture dishes at 37 degrees C. Cell sheets were harvested from the dishes after 2 weeks by reducing temperature to 20 degrees C. Histologic analyses, immunoblotting, and colony-forming assay were performed to characterize the cell sheets. Autologous transplantation was undertaken to reconstruct the corneal surfaces of rabbits with experimentally induced limbal stem cell deficiencies. RESULTS: Multilayered corneal epithelial sheets were harvested intact simply by reducing the temperature, without the use of proteases. Cell-cell junctions and extracellular matrix on the basal side of the sheet, critical to sheet integrity and function, remained intact. A viable population of corneal progenitor cells, close in number to that originally seeded, was found in the sheets. Harvested sheets were easily manipulated, transplantable without any carriers, and readily adhesive to corneal stroma so that suturing was not required. Corneal surface reconstruction in rabbits was highly successful. CONCLUSIONS: Cell sheet engineering technology allows us to create intact, transplantable corneal epithelial cell sheets that retain stem cells from limbal stem cells expanded ex vivo. Our research indicates highly promising clinical capabilities for our bioengineered corneal epithelial sheet.
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