Long-term efficacy and safety of ranibizumab administered pro re nata in Japanese patients with neovascular age-related macular degeneration in the EXTEND-I study.

PURPOSE: To evaluate the long-term efficacy and safety of ranibizumab administered pro re nata (PRN) in Japanese patients with choroidal neovascularization secondary to age-related macular degeneration during the extension phase of the EXTEND-I study. METHODS: EXTEND-I, an open-label, multicenter, Phase I / II study comprised: a single-injection (Group A); a multiple-injection (Groups A and B; the latter consisted of patients who did not participate in the single-injection phase); and an extension phase. In the extension phase, a PRN regimen of ranibizumab (0.3 or 0.5 mg) guided by monthly best-corrected visual acuity (BCVA) score and other ophthalmic examinations was employed. The efficacy variables included the mean BCVA change from Month 12 to the last visit in Group B. Safety was assessed in all patients. RESULTS: In the extension phase, efficacy was assessed only in Group B patients. The number of ranibizumab injections per year in the 0.3 and 0.5 mg Group B patients was 4.19 and 4.27, respectively. The mean BCVA change (SD) from Month 12 to the last visit was )3.6 (14.82) letters for 0.3 mg (n = 28) and )2.2 (7.92) letters for 0.5 mg groups (n = 33) in Group B. Conjunctival haemorrhage and nasopharyngitis were the most commonly reported adverse events. Of the 13 serious adverse events reported, cerebral infarction (two incidences) was suspected to be study-drug related. CONCLUSIONS: Pro re nata regimen of ranibizumab guided by monthly BCVA and other ophthalmic examinations appears effective in sustaining the BCVA gained with 12 monthly injections while reducing the number of injections during the extension phase. Ranibizumab was well tolerated during the extension phase.

Simultaneous intravitreal injection of triamcinolone acetonide and tissue plasminogen activator for central retinal vein occlusion: a pilot study.

PURPOSE: To evaluate the efficacy and safety of simultaneous intravitreal injection of triamcinolone acetonide (TA) and tissue plasminogen activator (tPA) for macular oedema associated with central retinal vein occlusion (CRVO). METHODS: Twenty eyes of 20 patients with CRVO were enrolled. A mixture of TA (4 mg) and tPA (25 µg) was injected into the vitreous of 20 eyes with CRVO. Best corrected visual acuity (BCVA) and macular thickness before and 1, 3, 6 and 12 months after the procedure were measured. RESULTS: The BCVA improved three lines or more in 65%, 55%, 55% and 53% of eyes and the mean macular thickness decreased from 1072 µm to 455, 450, 480 and 409 µm (p<0.001) at 1, 3, 6 and 12 months, respectively. Fifteen (75%) of the 20 eyes required at least one additional injection to prevent a recurrence of macular oedema. The intraocular pressure increased in four eyes. CONCLUSION: Overall, intravitreal injection of the TA/tPA mixture improved the BCVA by three lines or more in at least 50% of eyes and decreased the mean macular thickness at four time points without serious side effects. A randomised clinical trial is necessary to evaluate the efficacy of this treatment.

Activated protein C rescues the retina from ischemia-induced cell death.

PURPOSE: Ischemia causes severe and persistent visual loss in many eye diseases, including central retinal vein occlusion (CRVO) and diabetic retinopathy. Activated protein C (APC) has been demonstrated to reduce the cell death associated with ischemia in the brain and kidney. This study was performed to examine the ability of APC to rescue hypoxia-induced retinal cell death in vitro and in vivo. METHODS: Retinal pigment epithelium (RPE) and photoreceptor cells were placed in either a normoxic or a hypoxic chamber. Immediately before they were subjected to ischemia, the cultures were treated with APC (3-240 µg/mL). Incubation was followed by an MTT assay to determine the number of viable cells. The activity of caspase-3, -8, and -9 in RPE cells was also analyzed. Various concentrations of APC were intravitreally injected in a rat CRVO model, followed by TUNEL staining to detect the in vivo effects of APC. RESULTS: Lower concentrations of APC (0.3-30 µg/mL) showed a cell-protective effect against hypoxia in vitro, whereas higher concentrations (≥120 µg/mL) demonstrated cytotoxicity in both RPE and photoreceptor cells. Caspase-3, -8, and -9 were activated when the cells were exposed to hypoxia, but this activation was significantly inhibited by APC. Experimental CRVO-induced retinal cell apoptosis was reduced dramatically by intravitreal injection of APC. CONCLUSIONS: APC can reduce ischemia-induced cytotoxicity both in vitro and in vivo via blocking the activation of caspase-3, -8, and -9. APC may be a promising candidate for protecting the retina from ischemia.

New culture technique of human eliminable feeder-assisted target cell sheet production.

Cultured cell sheets for transplantation generally have been co-cultured with animal feeder cells, which carry risks because of different species and results in non-contact culture between the feeder and target cells. We developed a new technique to produce human eliminable feeder-assisted target cell sheets by novel human-derived genetically modified feeder cells. Three genes (human-derived telomerase reverse transcriptase gene, enhanced green fluorescent protein gene, and herpes simplex virus thymidine kinase gene) were transducted into human stromal cells, which enabled genetically modified feeder cells to be immortalized, labeled, and eliminated as needed. A target cell sheet was produced as one sheet by assisting the genetically modified feeder cells and successfully transplanted in vivo without their contamination. Genetically modified human eliminable feeder cells could be a promising tool for cultivated cell sheet transplantation.

Four cases of Acanthamoeba keratitis treated with phototherapeutic keratectomy.

PURPOSE: To report 4 cases of Acanthamoeba keratitis treated with excimer laser phototherapeutic keratectomy (PTK) and to discuss the clinical efficacy of this procedure. METHODS: Four cases with early stage Acanthamoeba keratitis resistant to medical amoebic therapy for at least 1 week and with an enlarged abscess underwent PTK. RESULTS: After PTK, the infected corneal lesions were removed and the clinical symptoms rapidly resolved in all cases. Another 40-µm ablation was required as a result of the 1-week delay in performing PTK. There was no recurrence during the postoperative period. CONCLUSION: When lesions are limited to about one third of the superficial corneal stromal layer, PTK could be the most beneficial option for treating Acanthamoeba keratitis, resistant to medical amoebic therapy using chlorhexidine or polyhexamethylene biguanide, because of direct removal of resistant amoebic cysts and better visual recovery without irregular astigmatism.

A novel method of culturing human oral mucosal epithelial cell sheet using post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction.

BACKGROUND/AIMS: To cultivate human oral mucosal epithelial cell sheets with post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction. METHODS: Human oral mucosal epithelial cells obtained from three healthy volunteers were cultured with x-ray-treated dermal fibroblasts (fibroblast group) and NIH/3T3 feeder layers (3T3 group) on temperature-responsive culture dishes. Media were supplemented using clinically approved products. Colony-forming efficiency was determined in both groups. Histological and immunohistochemical analyses were performed for cell sheets. Cell viability and purity of cell sheets were evaluated by flow cytometry. RESULTS: Colony-forming efficiency in the fibroblast group was similar to that in the 3T3 group. All cell sheets were well stratified and harvested successfully. The expression patterns of keratin 1, 3/76, 4, 10, 12, 13, 15, ZO-1 and MUC16 were equivalent in both groups. The percentage of p63-positive cells in the fibroblast group (46.1+/-4.2%) was significantly higher than that in the 3T3 group (30.7+/-7.6%) (p=0.038, t test). The cell viability and purity were similar between the two groups. CONCLUSION: This novel culture method using dermal fibroblasts and pharmaceutical agents provides a safe cell processing system without xenogenic feeder cells for ocular surface reconstruction.

Spectral-domain optical coherence tomographic findings in myopic foveoschisis.

PURPOSE: The purpose of this study was to investigate the sensitivity of various scan modes of spectral-domain optical coherence tomography (SD-OCT) for detecting pathologies that may accompany myopic foveoschisis, which is important for making surgical decisions and planning surgical strategies for myopic foveoschisis. METHODS: Twenty-one eyes of 18 patients were diagnosed with myopic foveoschisis by SD-OCT. Patients were examined with SD-OCT using both a five-line raster scan and a three-dimensional scan at the same visit. The detection rates of pathologies such as macular hole, epiretinal membrane, retinal vascular microfolds, internal limiting membrane detachment, paravascular microhole, and photoreceptor inner and outer segments defect were compared between modes. RESULTS: The three-dimensional scanning mode of the SD-OCT tended to be superior to both the one-line and five-line raster scanning modes for detecting inner and outer segments, epiretinal membrane, macular hole, paravascular microhole, and internal limiting membrane. The three-dimensional mode had a significantly higher detection rate (71%) of retinal vascular microfolds than both the 1-line (19%, P < 0.01) and 5-line raster scanning modes (33%, P < 0.05). CONCLUSION: The three-dimensional scan mode of SD-OCT was more sensitive for detecting the pathologies that accompany myopic foveoschisis, providing important information for vitreous surgery.

Ocular risk factors for choroidal neovascularization in pathologic myopia.

UNLABELLED: PURPOSE. To identify the risk factors for development of myopic choroidal neovascularization (mCNV), a major cause of visual impairment. METHODS. Enrolled in the study were 23 consecutive patients with bilateral high myopia (axial length, > or =26.5 mm or refractive error, < or =8 D) and unilateral newly developed mCNV who presented to the Myopia Clinic, Osaka University Hospital. Spectral-domain optical coherence tomography (SD-OCT) showed that the fellow eyes had a normal macula. The parameters in the affected and fellow eyes were compared between the individual patients, including best-corrected visual acuity (BCVA), intraocular pressure (IOP), refractive error, axial length, choroidal thickness (CT) (subfoveal, 1.5 mm superiorly and inferiorly), posterior staphyloma height 3 mm from the fovea, length of retinal pigment epithelium (RPE) curvature within 6 mm measured on SD-OCT images, and choroidal degeneration and lacquer crack formation, graded according to a published METHOD: RESULTS. The IOP, axial length, refractive error, and chorioretinal degeneration did not differ significantly. Affected eyes had a significantly higher lacquer crack grade (P < 0.05). The superior CT was not significantly different; the subfoveal and inferior CTs were significantly lower in the affected eyes (P < 0.05 and P < 0.001, respectively). The absolute value of the nasal posterior staphyloma height from the fovea was significantly greater in the affected eyes (P < 0.05), and the affected eyes had a significantly (P < 0.05) longer RPE curvature. CONCLUSIONS. Choroidal thinning resulting from increased RPE/choroid curvature is a risk factor for unilateral mCNV.

EXTEND-I: safety and efficacy of ranibizumab in Japanese patients with subfoveal choroidal neovascularization secondary to age-related macular degeneration.

PURPOSE: To evaluate the efficacy and safety of intravitreal ranibizumab for subfoveal choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) in Japanese patients. METHODS: This open-label, multicentre, Phase I/II study enroled patients into Group A (single injection of ranibizumab nonrandomized doses of 0.3 or 0.5 mg followed by 11 monthly injections of the same dose) and Group B (12 monthly injections of ranibizumab randomized to 0.3 or 0.5 mg). The primary efficacy endpoint was the mean change from baseline in best-corrected visual acuity (BCVA) score at Month 6. Safety was evaluated in all patients who received ranibizumab. RESULTS: Of 88 patients enroled, 12 entered Group A (six per dose) and 76 entered Group B (0.3 mg: n = 35; 0.5 mg: n = 41). Mean change from baseline in BCVA was significantly increased for both doses (Group B) at Month 6 (0.3 mg: +8.1 letters, p = 0.0006; 0.5 mg: +9.0 letters, p < 0.0001) and Month 12 (0.3 mg: +9.5 letters, p = 0.0001; 0.5 mg: +10.5 letters, p < 0.0001). At Month 12, one patient (0.3 mg) and 0 patients (0.5 mg) lost > or =15 letters, while 37.1% (0.3 mg) and 31.7% (0.5 mg) of patients gained > or =15 letters. Ocular serious adverse events (SAEs) of the study eye were reported in 1 and 2 patients in the 0.3- and 0.5-mg groups, respectively. Nonocular SAEs were experienced by 2 and 5 patients in the 0.3- and 0.5-mg groups, respectively. No cases of endophthalmitis were reported. CONCLUSION: Ranibizumab was effective and well tolerated in Japanese patients with subfoveal CNV secondary to AMD.

Efficacy of suprachoroidal-transretinal stimulation in a rabbit model of retinal degeneration.

Purpose. To develop a middle-sized animal model of outer retinal degeneration and to evaluate the effectiveness of suprachoroidal-transretinal stimulation (STS) in eliciting cortical potentials from this model. Methods. Twelve rabbits were intravenously injected with 0.47 mg/kg verteporfin and the retinas were irradiated with a red light for 90 minutes. Fluorescein angiography and full-field and focal electroretinography (ERG) were performed at 7 and 28 days after the irradiation. Electrically evoked potentials (EEPs) were elicited by electrical stimulation, with the STS electrode implanted over the irradiated region, 1 month and 1 year after the irradiation. EEPs were also recorded from three rabbits before and after retinotomy of the normal retina surrounding the degenerated area, to eliminate the influence of stray currents. The retina beneath the site of the STS electrode was examined histologically at 1 month (group 1) and 1 year (group 2) after the irradiation. Results. An extensive area of degeneration was detected histologically, mainly in the outer retina after the irradiation. Focal ERGs were not recorded when the stimulus was confined to the irradiated area; however, EEPs were successfully elicited by STS of the same area 1 month and 1 year after the irradiation. The 360 degrees retinectomy did not significantly alter the amplitudes, the implicit times, or the thresholds of EEPs evoked by STS. Conclusions. Verteporfin with light irradiation induces degeneration predominantly in the outer retinal layers in rabbits. The elicitation of EEPs by STS from the degenerated area suggests that the STS system may be useful in patients with retinitis pigmentosa.

Two cases of acanthamoeba keratitis diagnosed only by real-time polymerase chain reaction.

PURPOSE: To report 2 cases of Acanthamoeba keratitis whose causative pathogen was detected only by real-time polymerase chain reaction (PCR). METHODS: Histological examinations of corneal scrapings were stained with Fungiflora Y. Corneal scrapings were also cultured on nonnutrient agar. Real-time PCR analyses of corneal scrapings were also performed. RESULTS: Both cases had clinical signs and risk factors of Acanthamoeba infection. Histological examinations of corneal scrapings with Fungiflora Y staining were negative, and the cultures did not grow any pathogens. Real-time PCR analysis was positive for Acanthamoeba DNA from 2 corneal scrapings. Antiamoebic treatments led to excellent clinical improvements. CONCLUSIONS: These results indicate that PCR analyses can detect the DNA of Acanthamoeba in corneal scrapings and may be a valuable supplemental examination. This method is especially helpful when clinical signs and risk factors of Acanthamoeba infection are present but histological examinations with Fungiflora Y staining and cultures are negative.

A 27-gauge instrument system for transconjunctival sutureless microincision vitrectomy surgery.

OBJECTIVES: To evaluate the efficiency, preliminary safety, and feasibility of a 27-gauge instrument system for transconjunctival microincision vitrectomy surgery (MIVS) in a variety of vitreoretinal diseases. DESIGN: Experimental, interventional case series. PARTICIPANTS: Thirty-one eyes (31 patients) underwent a variety of vitreoretinal procedures using the 27-gauge transconjunctival MIVS system to treat epiretinal membrane (n = 10), idiopathic macular holes (n = 7), diabetic vitreous hemorrhage (n = 5), vitreous opacity with suspicion of intraocular lymphoma (n = 4), focal diabetic traction retinal detachment (n = 3), macular traction syndrome (n = 1), and macular edema secondary to central retinal vein occlusion (n = 1). METHODS: We developed a 27-gauge instrument system that includes an infusion line, a high-speed vitreous cutter, an illumination system, and a variety of vitreoretinal instruments, such as membrane forceps and sharp-tipped endophotocoagulation probes. The duty cycle of the 27- and 25-gauge cutters was measured for several cut rates using a high-speed imaging camera. Infusion and aspiration rates were measured using balanced saline solution (BSS) and porcine vitreous with different aspiration levels. Surgical outcomes, including anatomic success, visual outcomes, operating times, and intraoperative and postoperative complications, were evaluated. MAIN OUTCOME MEASURES: Duty cycle of cutters, infusion and aspiration rates, and surgical results of 27-gauge vitrectomy. RESULTS: Although the infusion and aspiration rates of the 27-gauge system measured in BSS were reduced to an average of 62% and 80%, respectively, compared with those of the 25-gauge system, the duty cycle of the 27-gauge cutter, 61% at 1000 cpm and 38% at 1500 cpm, was equal to or better than those of the 25-gauge cutter (62% and 28%, respectively). Analysis of the fluid dynamics showed that vented gas-forced infusion can be set to range from 20 to 30 mmHg to control intraocular pressure (IOP) during 27-gauge vitrectomy. Anatomic success was achieved in all study eyes (100%); 20 eyes (65%) had visual improvement of 3 lines or more. No eyes required conversion to larger gauge instrument. All sclerotomies self-sealed without hypotony (IOP < or = 7 mmHg) from 1 day postoperatively. CONCLUSIONS: Although the fluid dynamics and cutting efficiency of 27-gauge instruments are lower compared with 25-gauge MIVS, the 27-gauge system is feasible and may reduce concerns about wound sealing-related complications in selected cases. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Clinical findings in unilateral acute idiopathic maculopathy: new findings in acute idiopathic maculopathy.

We report a case of unilateral acute idiopathic maculopathy (UAIM) with new clinical findings. A 34-year-old Japanese man had a neurosensory retinal detachment (approximately 5 disk diameters) with yellowish-white exudates at the macula in the left eye (visual acuity (VA) 0.4). Fluorescein angiography (FA) showed early hypofluorescent spots and late pooling in the subretinal space. Three weeks after onset, indocyanine green angiography (IA) showed numerous hypofluorescent spots at the lesion. Optical coherence tomography (OCT) showed subretinal fluids and an elevated choroidal lesion with low reflectivity, suggesting choroidal edema. The VA and fundus appearance spontaneously resolved without treatment three months after onset. The VA was 1.0 six months after onset. Irregular pigmentation remained at the macular lesion. The main UAIM pathology may be outer retinal layer and retinal pigment epithelial inflammation. FA, IA, and OCT suggested that choroidal inflammation may be involved in the pathogenesis of UAIM.

Prevalence and features of keratitis with quantitative polymerase chain reaction positive for cytomegalovirus.

PURPOSE: To assess corneal scrapings and aqueous humor samples analyzed by polymerase chain reaction (PCR) that were positive for cytomegalovirus (CMV) in patients with keratitis of unknown origin and to investigate their clinical manifestations. DESIGN: Retrospective, interventional case series. PARTICIPANTS: Seventy-eight patients with epithelial (n=37), stromal (n=12), or endothelial keratitis (n=29) of unknown origin examined at the Osaka University Medical Hospital. METHODS: Clinical examination and tears, corneal scrapings, and aqueous humor specimens were evaluated by real-time PCR for CMV. MAIN OUTCOME MEASURES: Quantification of CMV DNA at the diagnosis of each type of keratitis with unknown origin and monitoring during the therapeutic course for CMV-positive cases. RESULTS: No cases of epithelial or stromal keratitis had CMV DNA. Seven of 29 corneal endotheliitis cases (24.1%) were positive for CMV. Cytomegalovirus-positive cases of corneal endotheliitis characterized by localized corneal edema and keratic precipitates included 4 patients who had undergone penetrating keratoplasty and were refractory to the treatment for graft rejection and 3 patients with idiopathic endotheliitis. Cytomegalovirus DNA copy numbers were estimated and ranged from 6.3x10(4) to 3.6x10(6)/ml. In all positive cases, the numbers of CMV DNA copies decreased within weeks during treatment with systemic and topical ganciclovir (GCV) combined with a topical steroid. Five eyes (62.5%) had clinical improvement. In cases of endothelial keratitis, diabetes mellitus was significantly higher in patients positive for CMV (71.4%) than in patients negative for CMV (18.2%, P=0.016, chi-square test). CONCLUSIONS: A total of 24.1% of cases with corneal edema of unknown origin were CMV positive and should be included in the differential diagnosis of idiopathic corneal endotheliitis or graft edema after penetrating keratoplasty, especially for bullous keratopathy. Real-time PCR for CMV, based on the diagnosis and monitoring of the clinical course, may be useful. Cytomegalovirus corneal endotheliitis requires early appropriate treatment using GCV. Because clinical remission after GCV may depend on the area of normal endothelium, early diagnosis and therapy are important for CMV corneal endotheliitis.

Artificial vision by direct optic nerve electrode (AV-DONE) implantation in a blind patient with retinitis pigmentosa.

The purpose of this study was to evaluate the efficacy and safety of artificial vision by using a direct optic nerve electrode (AV-DONE) in a blind patient with retinitis pigmentosa (RP). This device, comprising three wire electrodes (0.05 mm in diameter), was implanted into the optic disc of a patient with RP with no light perception vision and the device was left implanted. Six months later, visual sensations were elicited by electrical stimulation through each electrode and the thresholds for the phosphene perception elicited by pulses of 0.25-ms duration/phase and a pulse frequency of 320 Hz were 30, 5, and 70 microA for each electrode. The phosphenes, which ranged in size from that of a match head to an apple, were round, oval, or linear, primarily yellow, and focally distributed. The area of the phosphenes changed when the electrical stimulation was supplied from different electrodes. No complications arose during the follow-up period. Localized visual sensations were produced in a blind patient with advanced RP, suggesting that our system could lead to the development of a useful visual prosthesis system.