Vertical asymmetry of lamina cribrosa tilt angles using wide bandwidth, femtosecond mode-locked laser OCT; effect of myopia and glaucoma.

PURPOSE: Morphological features of the lamina cribrosa (LC) and optic disc may be important in the pathogenesis of glaucoma and myopic neuropathy. We therefore performed a cross-sectional study of patients with glaucoma and myopic neuropathy to evaluate vertical asymmetry of LC tilt angles (LCTAs) from Bruch's membrane opening (BMO). MATERIAL AND METHODS: Forty-six control eyes and 35 primary open-angle glaucoma (POAG) eyes were included. A raster scanning protocol with 300 single B-scans (without averaging) were obtained using optical coherence tomography with a wide-bandwidth, femtosecond mode-locked (ML) laser. Superior temporal to inferior nasal (ST) direction and inferior temporal to superior nasal (IT) direction (±45° rotation with a horizontal line) lines were drawn, and the angle between the inner edge of the BMO plane and the best fitting line for the anterior LC plane was measured as the LCTA. The generalized estimating equation was used to analyze the eye-derived data. RESULTS: Although no significant differences in either ST-LTCAs or IT-LTCAs were observed between the glaucoma group and non-glaucoma group, the IT-LCTAs were found to be significantly greater than the ST-LCTA in both the glaucoma and non-glaucoma groups (P < 0.001). After adjustment for other potential confounding factors by multivariate analysis, greater refractive errors were significantly correlated with IT-LCTAs. CONCLUSIONS: Vertical asymmetry of the LC tilting from the BMO plane exists in both normal and POAG eyes, and correlates with the degree of myopia.

Generation of broadband noise-like pulse from Yb-doped fiber laser ring cavity.

We have demonstrated a generation of the noise-like pulse (NLP) with broadband spectrum in a nonlinear polarized evolution-based passive mode-locked Yb-doped fiber (YDF) ring laser. At the cavity dispersion of near zero, the NLP with spectrum bandwidth up to 131 nm (FWHM) was obtained at a central wavelength of 1070 nm with output power of 136 mW and 80 MHz repetition rate. To our best knowledge, this spectrum bandwidth of NLP is the broadest among the reported YDF lasers. The autocorrelation function of pulse contained the short (30 fs) and long (4.6 ps) components. This short coherence light source is well suited for the optical coherent tomography used for ophthalmology at a wavelength of ∼1000 nm.

Three-dimensional optic nerve head images using optical coherence tomography with a broad bandwidth, femtosecond, and mode-locked laser.

PURPOSE: The aim of this study was to demonstrate the fine laminar structure of the optic nerve head (ONH), in vivo, using a broad wavelength, ultra-high resolution, and optically coherent tomography (OCT) system. METHODS: This high-resolution OCT system, based on a 200 nm bandwidth spectrometer and an 8 femtosecond ultra-short, mode-locked, coherent laser light source, enabled in vivo cross-sectional ONH imaging with 2.0 µm axial resolution. A total of 300 optic disc B-scans, which consisted of 300 × 2048 pixels, were obtained in 10 µm steps. Three-dimensional images were rendered from these images to obtain n face images of the optic disc. Fundus photography, scanning laser ophthalmoscopy (SLO), and standard OCT were also performed for all subjects. RESULTS: Thirty-six eyes of normal subjects and ten eyes of glaucoma patients with mean age of 40.0 ± 10.0 years were enrolled in this study. Sequential en face images, from the ONH surface to deeper layers, were reconstructed in 2.0 µm steps. Observation of the images indicated variations in the shape and arrangement of the lamina pores at different depths. Clear lamina pores were identified by this technique in 44 eyes, compared with the fundus camera (identified in six eyes), SLO (identified in 14 eyes), and standard OCT (identified in 24 eyes) (all comparisons, p < 0.001). CONCLUSIONS: The fine structure of the ONH could be resolved in vivo using our OCT, providing improved imaging that can be used in research and clinical applications for a better characterization of the anatomical and pathological features associated with glaucoma.

[Corneal subepithelial infiltrates that developed following intravitreous injection of bevacizumab].

BACKGROUND: Anti-vascular endothelial growth factor (anti-VEGF) therapies including intravitrous bevacizumab injection (IVB) emerged as an effective treatment for various fundus diseases. Reports of adverse reactions derived from bevacizumab itself are rare. We report a case in which corneal subepithelial infiltrates developed following IVB. CASE: A sixty-six-year-old man presented with visual acuity loss due to cystic macular edema secondary to branch retinal vein occlusion. He developed vascular congestion and subepithelial infiltrates in the peripheral cornea on day 2 following the first IVB. The findings resolved with topical steroid in twelve days. CONCLUSION: A case of subepithelial infiltrate in the peripheral cornea following IVB is reported. Since infiltrates were located along, and associated with the congestion of, the limbal vascular arcade, an innate immune response to bevacizumab may be indicated.

Montage images of spectral-domain optical coherence tomography in eyes with idiopathic macular holes.

PURPOSE: To describe the morphologic and anatomic relationships at the vitreoretinal interface, from the macula into the periphery, in patients with idiopathic macular hole. Montaged images of posterior and peripheral spectral-domain (SD) optical coherence tomography (OCT) studies were used to describe the anatomic vitreoretinal relationships. DESIGN: Prospective, consecutive, observational case series. PARTICIPANTS: Forty-six eyes of thirty-six consecutive patients with idiopathic macular hole and their fellow eyes. METHODS: Montage images of 4 radial OCT scans (horizontal, vertical, and 2 oblique scans) through the fovea were obtained in each case. MAIN OUTCOME MEASURES: Montage SD OCT images. RESULTS: In fellow eyes, potential precursor changes to macular hole revealed shallow perifoveal vitreous separation that extends peripherally toward the equator. Two distinct configurations were noted at the posterior vitreous face; eyes without holes had a smooth curvature, whereas eyes with holes were more likely to have wavy, folded, or scalloped vitreous surfaces. At the onset of separation, most posterior vitreous cortex had a smooth curvature, but posterior vitreous folds increased with progressive separation. Also notable were zones of double-layered retinoschisis in regions of adherent posterior vitreous. Resulting granular hyperreflection in the peripheral vitreous was detectable in 50% to 60% of stage 1 or 2 holes but in only 33% of stage 3 or 4 holes. CONCLUSIONS: The SD OCT montages taken at serial stages of idiopathic macular holes document distinct configurations of the posterior vitreous face, granular hyperreflection in the peripheral vitreous, and areas of peripheral retinoschisis. Montaging SD OCT images provides novel cross-sectional images of the vitreoretinal interface that may have broader application. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Associations of cigarette smoking but not serum fatty acids with age-related macular degeneration in a Japanese population.

PURPOSE: To assess modifiable environmental risk factors and protective factors for age-related macular degeneration (AMD) in a native Japanese population. DESIGN: A case-control study. PARTICIPANTS: We included 422 case-control samples composed of 279 consecutive AMD cases and 143 controls. METHODS: Information regarding systemic conditions and lifestyle were documented in each subject by standardized questionnaire including age, gender, smoking history, body mass index (BMI), and history of cardiovascular disease, hypertension, and diabetes. Serum fatty acids profiles were analyzed by gas chromatography performed on blood samples taken from each study participant. Logistic regression and multiple comparison analyses were utilized in this study. MAIN OUTCOME MEASURES: Population-specific information assessing systemic conditions, lifestyle, and serum fatty acid profiles. RESULTS: Among environmental factors analyzed cigarette smoking showed the most significant association with development of all AMD (P<0.00001; odds ratio [OR], 4.06; 95% confidence interval [CI], 2.22-7.43), typical neovascular AMD (P<0.0001, OR, 4.59; 95% CI, 2.29-9.18), and polypoidal choroidal vasculopathy (P<0.001; OR, 4.87; 95% CI, 1.96-12.1). Hypertension and BMI showed a mild association with AMD. Although male prevalence was significantly higher in all case groups than in controls with conventional Scheffe correction, there was no association of gender with AMD development when logistic regression analysis was used to adjust for cigarette smoking. There was no difference in fatty acid profiles, except for a mild association of eicosapentaenoic acid concentration in the all AMD group. CONCLUSIONS: In the Japanese population studied, cigarette smoking influenced the risk of AMD but fractionated serum fatty acid levels did not. Although prior reports indicate a male predominance in Japanese patients with AMD, this study demonstrates that cigarette smoking accounts for this confounding bias. In addition, our population-specific data do not demonstrate significant differences in serum fatty acid composition, including ω-3 and ω-6 long chain polyunsaturated fatty acids, in Japanese patients with and without AMD. These results are consistent with the high proportion of smokers in aged Japanese men and the high fish oil intake in this population. FINANCIAL DISCLOSURE(S): The authors have no proprietary or commercial interest in any of the materials discussed in this article.

Complement factor H and high-temperature requirement A-1 genotypes and treatment response of age-related macular degeneration.

PURPOSE: To determine whether there is an association between complement factor H (CFH), high-temperature requirement A-1 (HTRA1), vascular endothelial growth factor (VEGF), and pigment epithelium-derived factor (PEDF) genotypes and response to treatment with photodynamic therapy (PDT) for age-related macular degeneration (AMD) in a Japanese population. DESIGN: Prospective, case-control study. PARTICIPANTS: One hundred ten patients with exudative AMD treated by verteporfin PDT were recruited prospectively at the Department of Ophthalmology, Saitama Medical University Hospital, Saitama, Japan. METHODS: The patients were genotyped for 4 single nucleotide polymorphisms (SNPs; rs800292, rs1061170, rs1410996, rs2274700) in the CFH gene, a rs11200638-SNP in the HTRA1 gene, 3 SNPs (rs699947, rs1570360, rs2010963) in the VEGF gene, and 4 SNPs (rs12150053, rs12948385, rs9913583, rs1136287) in the PEDF gene using a TaqMan assay. MAIN OUTCOME MEASURES: The treatment outcomes and genotypes of CFH, HTRA1, VEGF, and PEDF polymorphisms. RESULTS: Best-corrected visual acuity 1 year after PDT was significantly increased in patients with the HTRA1-rs11200638 GG genotype as compared with patients with the GA or AA genotypes (P = 2.9 × 10⁻², 7.0 × 10⁻4, respectively). The rate of recurrence in the 12-month period after PDT was also associated with HTRA1-rs11200638 genotype (P = 3.12 × 10⁻²). Patients with the AA genotype of HTRA1-rs11200638 had an approximately 6-fold greater risk of the recurrence than patients with the GG genotype (P = 5.58 × 10⁻³). Significant differences were demonstrated in the mean time interval from the initial treatment to the time of recurrence for the genotypes of CFH-rs1410996/-rs2274700 (P = 8.50 × 10⁻³). CONCLUSIONS: The HTRA1-rs11200638 and CFH-rs1410996/-rs2274700 variants were associated with response to PDT in this study population. These variants may be used for genetic biomarkers to estimate visual outcomes and recurrences in the response to PDT with significant predictive power.

Phenotype and genotype characteristics of age-related macular degeneration in a Japanese population.

PURPOSE: To describe phenotype and genotype characteristics of age-related macular degeneration (AMD) in Japanese patients. DESIGN: A case-control study. PARTICIPANTS: A total of 550 case-control samples composed of 408 consecutive AMD cases and 142 controls. METHODS: Clinical information assessing age, gender, affected eyes, fundus features, and fluorescein/indocyanine green angiograms were systematically evaluated. Four single nucleotide polymorphisms (SNPs; rs800292, rs1061170, rs1410996, rs2274700) in the complement factor H (CFH) gene, 1 SNP (rs11200638) in the high-temperature requirement factor A1 (HTRA1) gene, 3 SNPs (rs699947, rs1570360, rs2010963) in the vascular endothelial growth factor (VEGF) gene, and 4 SNPs (rs12150053, rs12948385, rs9913583, rs1136287) in the pigment epithelium-derived factor (PEDF) gene were assessed using TaqMan technology. MAIN OUTCOME MEASURES: The clinical phenotype information and genotypes of CFH, HTRA1, VEGF, and PEDF polymorphisms. RESULTS: Of Japanese patients with neovascular AMD (nAMD), 219 (58.7%) had typical nAMD and 154 (41.3%) had polypoidal choroidal vasculopathy (PCV). The frequency of bilateral exudative involvement was similar between typical nAMD (15.5%) and PCV (13.6%) (P = 0.613). Significant soft drusen were observed in the fellow eyes of 88 (47.6%) of 185 patients with unilateral typical nAMD and in 25 (18.8%) of 133 patients with unilateral PCV (P = 1.24x10(-7)). A serous pigment epithelium detachment was seen in 55 (25.1%) of 219 patients with typical nAMD and in 64 (41.6%) of 154 patients with PCV. A significant association was noted in CFH-rs800292, CFH-rs1410996, CFH-rs2274700, and HTRA1-rs11200638 with AMD development (P = 2.36x10(-5), 7.18x10(-5), 7.18x10(-5), 2.70x10(-7), respectively; population attributable risk = 57.3%, 57.8%, 57.8%, and 58.9%, respectively). We estimated the highest-risk group to have an approximately 70-fold greater risk of nAMD compared with the lowest-risk group when analyzing a combination of 4 SNPs in the CFH and HTRA1 genes. CONCLUSIONS: The Japanese AMD phenotype is characterized by a higher frequency of PCV, male predominance, and lower frequency of bilateral presentation compared with Caucasian AMD. Genotype analyses demonstrate a significant population attributable risk for SNPs in the CFH and HTRA1 genes and demonstrate joint effects for both genes. Gene variants in both CFH and HTRA1 contribute significantly to the AMD phenotype in a Japanese population.

[Outcome of photodynamic therapy for exudative age-related macular degeneration with good visual acuity].

PURPOSE: To evaluate visual outcome one year after photodynamic therapy (PDT) in patients with exudative age-related macular degeneration (AMD) showing good visual acuity (VA). SUBJECTS AND METHODS: One hundred and thirteen patients with AMD who received PDT therapy were recruited for this study. The study subjects were divided into two groups : those having a better visual acuity than 0.6 (n = 34), and those with VA worse than 0.5 (n = 79). All patients were examined by corrected visual acuity, fundus biomicroscopy, and optical coherence tomography (OCT) before, 1 month, 3 months, and 12 months after PDT. RESULTS: Visual acuity at 12 months was either at baseline visual acuity or improved in 82% of the better vision group. Foveal retinal thickness was significantly reduced with PDT in both the better vision and the worse vision groups (p = 0.04, 0.008, respectively). The greatest linear dimension (GLD) was significantly associated with significant vision decline among baseline factors (p = 0.049)such as gender and age. CONCLUSIONS: PDT is an effective adjunct for AMD patients even when they have good baseline vision. To perform PDT safely for AMD in patients with good vision, both larger GLD and retinal pigment epithelial detachment have to be considered as a risk factor.

Photodynamic effects on retinal oxygen saturation, blood flow, and electrophysiological function in patients with neovascular age-related macular degeneration.

PURPOSE: To evaluate the effect of photodynamic therapy (PDT) on retinal functions, such as oxygen saturation, blood flow, and electrophysiological function using Fourier transform-based spectral retinal imaging (SRI), Heidelberg retinal flowmeter (HRF), and multifocal electroretinogram (mfERG). METHODS: This was a prospective interventional case series. Twenty-two patients with age-related macular degeneration (AMD) with unilateral choroidal neovascularization (CNV) were examined using SRI, HRF, and mfERG before and 1 week and 1 month after PDT. Eleven fellow eyes without CNV and 11 control eyes of 11 age-matched healthy volunteers were also investigated. Eleven of 22 patients with neovascular AMD were retreated using verteporfin PDT and examined using a protocol similar to the one used for the first treatment. RESULTS: Oxygen saturation levels in the macula of eyes with neovascular AMD were significantly higher than those in normal control eyes (P = 0.026) but were not significantly higher in eyes with nonneovascular AMD. Oxygen saturation levels decreased 1 week after a single treatment (P = 4.59 x 10(-3)) and retreatment (P = 0.0134) and recovered to baseline levels at 1 month follow-up (P > 0.05). HRF demonstrated reduced mean blood flow at 1 week after single treatment (P = 9.22 x 10(-4)) and retreatment (P = 0.0307). One month after PDT, mean blood flow tended to show recovery. There was a similar decrease in mfERG amplitude 1 week after treatment, but the logarithm of minimum angular resolution (logMAR) vision was stable or improved throughout follow-up. CONCLUSION: Oxygen saturation levels, mean blood flow, and mfERG amplitude decreased 1 week after PDT treatment in both single treatment and retreatment groups. Although logMAR vision is stable or improved, our data showed transient functional deterioration in the retina after PDT treatment.

Coding and noncoding variants in the CFH gene and cigarette smoking influence the risk of age-related macular degeneration in a Japanese population.

PURPOSE: Ethnic variation has been reported in age-related macular degeneration (AMD)-associated Y402H polymorphism in complement factor H (CFH). This variation is evident in the Japanese population. Recently a strong association between a novel single-nucleotide polymorphism (SNP; rs1410996) in the CFH gene and AMD has been identified in Caucasian patients. The present study was undertaken to investigate whether four coding and noncoding variants of the CFH gene, including rs1410996, are associated with AMD in native, unrelated Japanese patients. METHODS: A total of 188 patients with AMD and 139 control subjects without AMD were recruited for the study. Four SNPs (rs800292, rs1061170, rs1410996, and rs2274700) in the CFH gene were assessed by genotyping assay. The information regarding systemic conditions and lifestyle including smoking were documented in each subject by standardized questionnaire. RESULTS: The intronic SNP (rs1410996) and the synonymous SNP (rs2274700) were associated with a significant risk of AMD (P = 2.37 x 10(-5) and 3.52 x 10(-5), respectively). A significant association was also noted between a coding variant (rs800292, I62V) and AMD (P = 8.63 x 10(-6)). In contrast, the Y402H variant showed no significant association with AMD (P = 0.101). Two common haplotypes also demonstrated significant association with AMD (P = 1.08 x 10(-3) and 2.00 x 10(-5)). Among the environmental factors, smoking alone had a significant association with AMD (P = 1.17 x 10(-4)). CONCLUSIONS: Although the Y402H variant was not significantly associated with AMD, other coding and noncoding variants in the CFH gene including rs1410996 and smoking moderately influenced the risk of AMD in a Japanese population.

Treatment of stage 2 macular hole by intravitreous injection of expansile gas and induction of posterior vitreous detachment.

PURPOSE: To demonstrate the morphological and functional outcomes of intravitreous injection of an expansile gas bubble for the treatment of stage 2 macular holes. DESIGN: Prospective interventional case series. PARTICIPANTS: Twenty eyes of 20 consecutive patients with unilateral stage 2 macular holes. METHODS: The patients underwent intravitreous sulfur hexafluoride injection, followed by postoperative facedown positioning for 3 to 5 days. The patients were observed per protocol schedule with complete ophthalmological examination, including determination of corrected visual acuity (VA), slit-lamp biomicroscopic examination, funduscopic examination, and optical coherence tomography (OCT). Follow-up was greater than 12 months for all patients (mean, 19.5). MAIN OUTCOME MEASURES: Posterior vitreous detachment, anatomical closure of the hole, VA, and hole diameter as measured by OCT. RESULTS: Detachment of the posterior vitreous at the macula was achieved in 19 of 20 eyes (95%). Ten cases (50%) had anatomical closure of the hole with intravitreous gas injection alone. The remaining 10 cases (50%) achieved anatomical closure of the hole after subsequent vitreous surgery. There was a significant difference in hole diameter (P = 0.004) and in pretreatment vision (P = 4.5x10(-5)) in patients for whom gas alone resulted in hole closure and those proceeding to vitreous surgery. Hole closure by gas injection alone was achieved in 7 of 7 eyes (100%) with pretreatment vision better than 20/40 and in 6 of 7 eyes (86%) when the hole was smaller than 200 mum. There were no major complications in this series of patients. Successfully treated macular holes remained closed at all follow-up points during the study period. CONCLUSION: An intravitreous injection of an expansile concentration of the inert sulfur hexafluoride gas alone frequently induces detachment of the posterior vitreous in the aged eye. Anatomical closure of the hole without major complications is more likely in smaller holes with better pretreatment vision. This technique may have clinical application for stage 2 holes in selected cases. A number of potential advantages including decreased morbidity and a potential cost savings may result from successful utilization of this procedure.

Intraocular biodistribution of mono-L-aspartyl chlorin e6 in a primate choroidal neovascularization model.

BACKGROUND AND OBJECTIVE: To describe the biodistribution of a hydrophilic sensitizer, mono-L-aspartyl chlorin e6 (NPe6), in a primate model of choroidal neovascularization (CNV). MATERIALS AND METHODS: NPe6 (25 mg/kg) and indocyanine green (ICG, 15 mg/kg) were intravenously administered simultaneously in monkeys with laser-induced CNVs. Eyes were enucleated and examined by fluorescence microscopy immediately, 20 minutes, and 1, 4, and 24 hours after dye injection. RESULTS: Fluorescence peaked in retinal vessels immediately after dye administration; rapid washout began 1 hour postinjection. In contrast, experimental CNV demonstrated little fluorescence immediately after injection, with increased intensity at later time points. Peak dye fluorescence in CNV occurred 1 hour following dye injection. NPe6 fluorescence diminished rapidly and washed out completely by 24 hours. The dye accumulation and retention pattern of ICG dye resembled that of NPe6. CONCLUSIONS: This study demonstrated the selective accumulation of NPe6 photosensitizer and ICG dye within experimental CNV and rapid clearance from the circulation and retinochoroidal tissue.

[Case of corneally displaced malignant conjunctival melanoma].

BACKGROUND: We studied the clinicopathologic characteristics in a patient with malignant conjunctival melanoma associated with corneal invasion. CASE: A 62-year-old man had a small melanocytic lesion of the inferior palpebral conjunctiva. Previously he had undergone excisional biopsy and was diagnosed as having melanocytic hyperplasia without cytological atypia at an other hospital. He developed recurrence and was referred to us. The pigmentary lesion was observed in the entire inferior palpebral conjunctiva. Biomicroscopic examination revealed that there was a granular pigment lesion in the cornea. The patient was diagnosed as having conjunctival melanoma with corneal invasion and treated with orbital exenteration and chemotherapy in our hospital. Clinicopathologic tests revealed malignant melanoma cells invading through the bulbar conjunctiva and into the cornea. Ultrastructural study by electron microscopy of the pigmented tumor cells in the cornea showed several lobations of the nuclei, a large active-appearing nucleolus, and an aberrant granular melanosomal morphology. CONCLUSIONS: The infiltration of palpebral malignant conjunctival melanoma was limited to the epidermis of the cornea.

Glaucomatous changes in lamina pores shape within the lamina cribrosa using wide bandwidth, femtosecond mode-locked laser OCT.

PURPOSE: The lamina cribrosa (LC) is known to play a critical role in the pathogenesis of glaucoma. Although it has been reported that striae-shaped or slit-shaped lamina pores are more frequent in eyes with primary open angle glaucoma (POAG), this observation is based only on fundus photography. The primary object of this study is to perform layer-by-layer comparisons of the shape of lamina pores within the LC in vivo. DESIGN: Cross-sectional study. METHODS: Optic nerve head B-scans were obtained using custom-made broad-wavelength optical coherence tomography with a mode-locked laser. A total of 300 single B-scans per eye were obtained and three-dimensional images were rendered from these image sequences to obtain 2-µm thin-slice en face images of the LC. Elongation indices (EIs) of the lamina pores were measured from the anterior surface (AS) of the LC to the deeper layers in 40-µm increments. RESULTS: Thirteen eyes from 10 primary open angle glaucoma (POAG) patients of mean deviation -15.2 (-16.5, -12.9) (median [25,75 percentile]) dB and 10 eyes from 7 normal controls were studied. Although the EI value was not significantly different between the superior, temporal and inferior regions of the LC at any depth level in either group, it was greater at the AS than at the 40 µm and 80 µm depth levels (P < .001) in both groups, and was greater in the POAG group only at the AS and 40 µm depth level (P ≤ .05). After adjustment for age and refraction, the effects of depth and presence of POAG on the EI value remained significant. Also, the severity of glaucoma and depth were significant factors associated with EI in multivariate analysis. CONCLUSIONS: Elongation of lamina pores was significantly more evident at the anterior surface and the 40-µm depth level of the LC in POAG eyes than in normal eyes, suggesting that nerve fiber bundles passing through the LC were under greater stress in the anterior layers of the LC.

Transpupillary thermotherapy-induced modification of angiogenesis- and coagulation-related gene expression in the rat posterior fundus.

PURPOSE: To study gene expression changes in the rat retina and choroid following transpupillary thermotherapy (TTT) and to identify molecular mechanisms that may enhance treatment of choroidal neovascularization, complicating age-related macular degeneration. METHODS: One fundus of Brown Norway rats was treated with an 810 nm diode laser while the contralateral fundus received no treatment. The mRNA was extracted and processed for cDNA microarray analysis. Genes with increased expression were validated by semiquantitative reverse transcription polymerase chain reaction (PCR) and quantitative real-time PCR (qRT-PCR). RESULTS: Of the 14,815 cDNA elements on the array, 12 genes were up-regulated in TTT treated eyes. Upregulation of eight of these 12 genes could be verified by semiquantitative RT-PCR. The eight verified genes were EPCR, IL-1beta, MCP-1, TSP-1, Fgl, Asns, MT-2, and NMDMC, which included 4 angiogenesis- and coagulation-related genes. CONCLUSIONS: This study demonstrates upregulation of angiogenesis- and coagulation-related genes following TTT. The response profile and its temporal relationships provide insight into the molecular mechanisms that lead to vascular occlusion and antiangiogenesis induced by TTT.

[Ultra-late phase of indocyanine green angiography in a case with metastatic choroidal tumor].

PURPOSE: To report a case with metastatic choroidal tumor evaluated with ultra-late phase of indocyanine green (ICG) angiography. CASES: A 43-year-old woman had metastatic choroidal tumor originating from the thymus with an associated serous retinal detachment. ICG video-angiography demonstrated the tumor as hypofluorescent during the arterio-venous phase and hyperfluorescent during the conventional late phase (about 30 minutes after dye injection), and in the ultra-late phase (24 hours after dye injection). The original and metastatic tumors were treated by radiotherapy, resulting in flattening of the choroidal tumor and resolution of the serous retinal detachment. After the treatment, ICG angiography showed decay of ICG fluorescence and no apparent late accumulation of residual ICG in the tumor. CONCLUSION: In a case with metastatic choroidal tumor, ICG dye accumulated in the tumor tissue before the treatment, but it decreased after treatment. This finding suggested that dye accumulation may be correlated with the tumor activity and requires a further study, enrolling patients with metastatic choroidal tumor for ICG angiographic evaluation.

[Melanoma-associated retinopathy with unknown primary site in a Japanese woman].

BACKGROUND: We report the clinical features of the first case of a Japanese person with melanoma-associated retinopathy. CASE: A 44-year-old woman complained of photopsia and blurred vision in her right eye, and was treated with steroids for uveitis by an ophthalmologist. She was referred to our hospital for further examination. After one month of treatment, she still complained of photopsia in her right eye. The best corrected visual acuity in the right eye was 0.8 and these was sensitivity loss in the central visual field test. Ophthalmoscopy and fluorescein angiography showed some retinal vasculitis in the right eye. A full-field electroretinogram demonstrated a negative-type electroretinogram (ERG) waveform with attenuation of the b-wave amplitude in the right eye. A dark adaptation test revealed sensitivity loss of the rods. The lymph nodes on the right side of her neck were examined and the diagnosis was made of metastic cutaneous melanoma with unknown primary site; her visual dysfunction was diagnosed as melanoma-associated retinopathy. The retinal inflammation improved after steroid treatment, but her visual dysfunction remained. Chemotherapy and an immunotherapy regimen was begun, but 36 months later she died of metastatic melanoma in the lungs. CONCLUSIONS: A woman treated for uveitis without any prior systemic and ocular diseases was diagnosed with melanoma-associated retinopathy and metastatic melanoma in the cervical lymph nodes of unknown primary origin. The first ocular symptoms were photopsia and blurred vision, not night blindness. ERG was useful for diagnosing this rare ocular condition in an early stage.

Laser photocoagulation and, to a lesser extent, photodynamic therapy target and enhance adenovirus vector-mediated gene transfer in the rat retina.

PURPOSE: To evaluate the transduction efficiency and localization of a reporter gene after intravitreous injection of adenovirus vector in laser photocoagulation (PC)- and photodynamic therapy (PDT)-treated eyes. METHODS: Adult Lewis rats received fundus PC, fundus PDT, or no treatment. Intravitreous injection of an adenovirus vector containing the construct expressing beta-galactosidase (AdlacZ.11D) was performed in each group. All eyes were then enucleated for histochemistry and processed for quantitative image analysis. RESULTS: In eyes with no treatment, there was moderate to intense staining for lacZ in the anterior segment, but little in the retina. In eyes treated with PC and PDT, there was significantly more LacZ staining in the retina. The increased staining corresponded closely with the sites treated with PC and PDT. Gene transduction in PC-treated eyes was enhanced and extended to at least 135 days after virus delivery, but not extended in PDT-treated eyes. Gene transfer and expression were targeted and enhanced at the site of laser burns, at all doses tested (3 x 10(5) to 3 x 10(9) particles per eye). CONCLUSIONS: Compared with untreated eyes, eyes treated with PC and to a lesser extent PDT, manifest increased transduction efficiency, in areas of the retina that are targeted by laser treatment. This finding suggests a new and promising strategy for the treatment of retinochoroidal neovascularization. Adenovirus gene therapy in combination with PC or PDT would have the advantage of increased transduction efficiency; increased duration of transgene expression; targeted delivery; and, potentially, a lower effective dose of virus.

Endothelial nitric oxide synthase gene is associated with diabetic macular edema in type 2 diabetes.

OBJECTIVE: We examined the endothelial nitric oxide (eNOS) gene polymorphisms to assess its possible association with diabetic retinopathy and macular edema. RESEARCH DESIGN AND METHODS: A total of 226 patients with type 2 diabetes and 186 healthy subjects were studied. Type 2 diabetic patients consisted of 110 patients without retinopathy, 46 patients with nonproliferative diabetic retinopathy, and 71 patients with proliferative diabetic retinopathy. Diabetic macular edema was present in 48 patients. Three polymorphisms of the eNOS gene were determined: T-786C in the promoter region, 27-bp repeat in intron 4, and Glu298Asp in exon 7. RESULTS: Close linkage disequilibrium was observed between the T-786C polymorphism and the 27-bp repeat, as has been previously reported, but Glu298Asp was not in linkage disequilibrium with the other two polymorphisms. The eNOS gene polymorphisms were not significantly associated with the presence of retinopathy or with retinopathy severity or type 2 diabetes itself. However, by both association study and multiple logistic regression analysis, the T-786C and 27-bp repeat polymorphisms were significantly associated with a risk of developing macular edema with the -786C allele and the "a" allele increasing the risk. CONCLUSIONS: The present study suggests that the eNOS gene is a novel genetic risk factor for diabetic macular edema. The eNOS gene polymorphisms may contribute to the development of macular edema by impairing basal eNOS expression and resulting in the breakdown of the blood-retina barrier.