Assessment of central vision and macular structure in patients undergoing iodine-125 brachytherapy for ciliochoroidal melanoma.

OBJECTIVES: To prospectively report standardized visual function and macular structural assessment in patients undergoing iodine-125 brachytherapy for choroidal and ciliary body melanoma. MATERIALS AND METHODS: Patients were enrolled for pretreatment and annual posttreatment assessment. Evaluations included ophthalmic history; standardized refraction; visual acuity, contrast sensitivity, and color vision measurement; comprehensive ophthalmic examination; fundus photography; fluorescein angiography; optical coherence tomography; and ultrasonography. Radiation doses to the foveola and optic disc margin were calculated. RESULTS: Forty-two patients were enrolled. Melanoma location included 3 in the ciliary body, 7 anterior, 11 equatorial, 13 posterior, and 8 macular tumors. Mean apical tumor height was 4.45 mm (range 1.79-9.83 mm) and mean longitudinal tumor diameter was 9.41 mm (range 4.52-4.73 mm). Pretreatment mean best-corrected Ferris-Bailey early treatment diabetic retinopathy study visual acuity was 50 (standard deviation +/- 15) letters (Snellen equivalent 20/32, range 20/15 to hand motions). The mean Pelli-Robson contrast threshold percentage was 4.1% (+/- 2.5%). The mean Hardy-Rand-Rittler color vision score was 13/14 (+/- 2.7). Mean distances from the posterior edge of the tumor to the foveola and the optic disc margin were 6.99 mm (+/- 6.22 mm) and 7.28 mm (+/- 5.98 mm), respectively. At the foveola, median total radiation dose was 36.2 Gy (+/-50.6 Gy) and median dose rate was 31.6 cGy/h (+/- 39.8 cGy/h). At the optic nerve, median total radiation dose was 42.8 Gy (+/- 30.8 Gy) and median dose rate was 36.2 cGy/h (+/- 21.4 cGy/h). CONCLUSION: This prospective assessment of macular structure and function will provide more complete understanding of the ocular effects of radiation therapy for ocular melanoma.

Transscleral fine-needle aspiration biopsy of macular choroidal melanoma.

PURPOSE: To report transscleral 30-gauge fine-needle aspiration biopsy (FNAB) for cytology and cytogenetics in eyes with macular choroidal melanoma. DESIGN: Prospective, interventional case series. METHODS: Twenty-five patients (25 eyes) who underwent transscleral 30-gauge FNAB of macular choroidal melanoma immediately prior to iodine-125 plaque placement were included in this study, conducted at a tertiary care university hospital. The main outcome measures were FNAB feasibility, cytology, cytogenetic analysis for monosomy 3, and surgical complications. RESULTS: Transscleral 30-gauge FNAB of choroidal melanoma in the macula was performed in 24 of 25 (96%) eyes and was not feasible owing to insufficient exposure in one eye (4%). Biopsy was diagnostic of choroidal melanoma in 17 of 24 (71%) eyes. Fluorescent in situ hybridization (FISH) and/or GeneChip 500k NspI Mapping array (Affymetrix, Santa Clara, California, USA) analysis for monosomy 3 was completed in 16 of 24 (67%) revealing monosomy 3 in five eyes and disomy 3 in 11 eyes. Retinal perforation (four eyes) did not require treatment or result in retinal detachment; submacular hemorrhage (nine eyes) and vitreous hemorrhage (five eyes) cleared spontaneously within one month. CONCLUSION: Transscleral FNAB of macular choroidal melanoma is feasible in most eyes and frequently yields cytogenetic information relevant to prognosis.

Photodynamic therapy induces caspase-dependent apoptosis in rat CNV model.

PURPOSE: To investigate the mechanism of cell death in laser-induced choroidal neovascularization (CNV) after photodynamic therapy (PDT). METHODS: PDT was performed in Brown-Norway rats using laser light at a wavelength of 689 nm, irradiance of 600 mW/cm(2), and fluence of 25 J/cm(2) after intravenous injection of verteporfin at the doses of 3, 6, and 12 mg/m(2). Apoptotic cells in CNV were detected by TUNEL assay at 1, 3, 6, 15, 24, and 48 hours after PDT. Caspase activation at 1, 3, 6, 15, and 24 hours after PDT was determined by immunohistochemistry (IHC) with a cleaved caspase-3 or -9 antibody. Akt activity was determined by Western blot and IHC with a phosphorylated-Akt (pAkt) antibody. To investigate the roles of Akt in PDT-induced apoptosis, insulin-like growth factor (IGF)-1, an Akt activator, with or without wortmannin, an inhibitor of PI3K-Akt pathway, was injected into the vitreous before PDT. RESULTS: The number of TUNEL-positive cells in CNV increased at 3 hours after PDT and peaked at 6 hours, showing a dose dependence of verteporfin. Caspase activation was detected in TUNEL-positive cells. Dephosphorylation of Akt in CNV occurred within 1 hour. IGF-1 significantly activated Akt and suppressed the number of TUNEL-positive cells in CNV, and the effects of IGF-1 were diminished by wortmannin. CONCLUSIONS: PDT induced caspase-dependent apoptosis in CNV. These results suggest that PDT leads to dephosphorylation of Akt and subsequent activation of the caspase-dependent pathway. Understanding the intracellular signaling mechanisms of apoptosis in PDT may lead to more selective and effective treatment of CNV secondary to age-related macular degeneration.

Spontaneous macular hole closure in bilateral macular holes.

The natural course of full thickness macular hole is progression in size and stage. There have been reports of spontaneous closure of unilateral idiopathic full thickness macular holes, but we report the first case of spontaneous closure of a full thickness macular hole in one eye in a patient with bilateral idiopathic full thickness macular holes. After macular hole surgery in the left eye of the patient, spontaneous closure of the macular hole in the right eye was observed during the follow-up period.

Treatment considerations for primary uveal melanoma with choroidal metastasis to the fellow eye.

We report two cases of primary uveal melanoma with metastatic involvement of the contralateral eye. Two female patients presented with primary choroidal melanoma. In the first case, primary enucleation of the affected eye was performed. Two years later, systemic tumor spread with contralateral choroidal melanoma was detected. A decision for observation of the ocular metastasis was made. In the second case, systemic tumor spread was already evident at time of initial diagnosis of the ocular melanoma. Six months later, a choroidal metastasis was detected in the fellow eye. Again, observation was recommended. In conclusion, systemic spread of primary choroidal melanoma may include a choroidal metastasis to the contralateral eye. Observation of the second affected eye may be prescribed.

Optos Panoramic200A fluorescein angiography for proliferative diabetic retinopathy with asteroid hyalosis.

We report a unique situation in which the use of Optos Panoramic200A fluorescein angiography directed the management of a patient with asteroid hyalosis and active proliferative diabetic retinopathy. The Optos Panoramic200A system provides a 200 degrees field of view, which may be useful in selected cases where simultaneous view of the posterior pole and periphery is important. The Optos fluorescein angiogram directed the management of our patient with proliferative diabetic retinopathy combined with asteroid hyalosis.

Reduced photoreceptor damage after photodynamic therapy through blockade of nitric oxide synthase in a model of choroidal neovascularization.

PURPOSE: To investigate the role of nitric oxide synthase (NOS) in photoreceptor degeneration associated with photodynamic therapy (PDT) in a laser-induced model of choroidal neovascularization (CNV). METHODS: PDT was performed in monkey and Brown Norway rats with laser-induced CNV. L-NAME, a NOS inhibitor, or saline was injected intraperitoneally in rats with CNV. An NO donor, or saline, was injected intravitreously into normal rats. Photoreceptor apoptosis was evaluated by TUNEL and electron microscopy. NOS, ED-1, and cleaved-caspase-3 (c-casp-3) expression were determined by immunohistochemistry. CNV lesions were examined by fluorescence angiography and choroidal flat mount. RESULTS: TUNEL and electron microscopy showed photoreceptor apoptosis after PDT. In rats, there were significantly more TUNEL-positive cells in the photoreceptors 24 hours after PDT, whereas in the CNV lesions there were more TUNEL-positive cells 6 hours after PDT. C-casp-3 was detected in the CNV lesions but not in the photoreceptors after PDT. There was no difference in the numbers of ED-1-positive macrophages before and after PDT. However, inducible NOS (iNOS) was increased after PDT in macrophages. Intravitreous injection of the NO donor without PDT also induced substantial photoreceptor apoptosis. L-NAME-treated animals had significantly fewer TUNEL-positive cells in the photoreceptors than saline-treated animals after PDT (P < 0.05). There were no differences in CNV size and leakage between L-NAME- and saline-treated groups. CONCLUSIONS: iNOS expression in macrophages contributes to PDT-induced photoreceptor degeneration. NOS inhibition reduces PDT-induced photoreceptor degeneration without compromising the treatment effect of PDT in an experimental model of CNV.

Surveillance of the eye and vision in clinical trials of CP-675,206 for metastatic melanoma.

PURPOSE: To determine the ocular safety of CP-675,206 (Pfizer, New York, New York, USA), a fully human anti-cytotoxic T lymphocyte-associated antigen 4 monoclonal antibody in clinical trials of immunotherapy of metastatic melanoma. DESIGN: Prospective, nonrandomized study of the eye and vision in phase I/II clinical trials of CP-675,206 in metastatic melanoma conducted at the University of California, Los Angeles. METHODS: Patients with regional or distant metastatic melanoma were enrolled in phase I/II clinical trials evaluating the safety and antitumor efficacy of CP-675,206 alone or in combination with melanoma antigen peptide-pulsed dendritic cell vaccines. Ophthalmic evaluation was performed at the onset of CP-675,206 immunotherapy (baseline evaluation), two months or more after the onset of CP-675,206 immunotherapy (end-study evaluation), and at two- to three-month intervals thereafter in patients who continued to receive CP-675,206 immunotherapy (poststudy evaluation). Baseline and end-study evaluations included comprehensive ophthalmic examination, psychophysical and electrophysiologic visual function assessment, fundus photography, fluorescein angiography, and visual function assessment. RESULTS: Twenty patients with metastatic melanoma arising from the skin, mucosa, eye, or unknown site were evaluated. Systemic toxicity attributed to CP-675,206 included dermatologic manifestations, diarrhea, and autoimmune hepatitis with panhypopituitarism. A subset of patients receiving CP-675,206 demonstrated antitumor efficacy with partial response or complete response of metastatic melanoma. Comparison of ophthalmic baseline with end-study evaluations in all 20 patients and limited-term poststudy evaluations showed no adverse effect of CP-675,206 immunotherapy on the eye or vision. CONCLUSIONS: In this study, CP-675,206 immunotherapy for metastatic melanoma did not adversely affect the eye or vision.

Fluorescent in situ hybridization for monosomy 3 via 30-gauge fine-needle aspiration biopsy of choroidal melanoma in vivo.

OBJECTIVE: To report the feasibility of intraoperative transscleral fine-needle aspiration biopsy at plaque surgery to obtain cells for monosomy 3 analysis in patients with choroidal melanoma. DESIGN: Consecutive interventional case series. PARTICIPANTS: Eighteen patients (18 eyes) with choroidal melanoma who had fine-needle aspiration biopsy performed with a 30-gauge needle at time of iodine 125 plaque placement. INTERVENTION: Cytology and cytogenetic analysis for monosomy 3 were obtained from biopsy specimens. MAIN OUTCOME MEASURES: Cytology, cytogenetic analysis for monosomy 3, and complications and feasibility of transscleral fine-needle aspiration biopsy of choroidal melanoma in vivo. RESULTS: Fine-needle aspiration biopsy was diagnostic of choroidal melanoma in 14 of 18 cases and resulted in viable cell cultures for fluorescent in situ hybridization (FISH) analysis in 9 cases. Fluorescent in situ hybridization for monosomy 3 was positive in 4 of the 9 cases. One patient had a mild vitreous hemorrhage. Tumors between 2 and 3 mm in height and those that yielded cells that did not attach in culture were most likely to have insufficient growth for FISH analysis. CONCLUSIONS: Transscleral fine-needle aspiration biopsy and FISH for monosomy 3 may provide important prognostic information on patients who undergo plaque radiotherapy for choroidal melanoma.

High-density genome array is superior to fluorescence in-situ hybridization analysis of monosomy 3 in choroidal melanoma fine needle aspiration biopsy.

PURPOSE: Using fluorescence in situ hybridization (FISH) and high-density single nucleotide polymorphism (SNP) mapping genome array, we comparatively evaluated chromosome 3 status and other chromosomal aberrations within a series of choroidal melanomas biopsied by fine needle aspiration (FNAB). METHODS: Transscleral FNAB was performed in 59 patients (59 eyes) who had a clinical diagnosis of choroidal melanoma. Biopsies were processed for chromosome 3 status by centromeric interphase FISH, cytopathology, cell culture, and simultaneous genomic DNA and RNA mapping array analysis. RESULTS: FISH yielded chromosome 3 status in 38 of 59 (64%) eyes, while high-density SNP mapping array yielded chromosome 3 status in 43 of 59 (73%) eyes. Monosomy 3 was detected by FISH in 15 of 38 (39%) cases, and high-density SNP mapping array data confirmed the finding in 13 of the 15 cases. Furthermore, high-density SNP mapping array revealed five additional cases of significant chromosome 3 aberration not detected by FISH. High-density genomic mapping also provided detailed patterns of chromosomal gain and loss on chromosomes 1, 6, 8, and 9 which segregated into two groups characterized by either monosomy 3 or chromosome 6p gain. CONCLUSIONS: High-density SNP mapping array was better than FISH in detecting chromosome 3 aberrations and monosomy in our melanoma samples. More importantly, the mapping arrays detected additional patterns of chromosomal aberration, which suggest specific pathways for cytogenetic rearrangements in choroidal melanoma and may improve prognostic testing.

Age-related macular degeneration: a practical approach to a challenging disease.

Age-related macular degeneration (AMD) is the leading cause of blindness in older North Americans. The clinical spectrum, risk factors, pathophysiology, and potential therapeutic options for AMD warrant a careful review. Despite the growth in treatment options for this disease, there is no current curative therapy. Of critical importance is attention to modifiable risk factors--improvements in cardiovascular status, including smoking cessation, and routine ophthalmic monitoring for opportunities to provide early intervention. In addition, a low-vision assessment to investigate the potential use of visual assistive devices may be beneficial to any patient who has experienced a decrease in vision. Finally, education regarding the clinical course of age-related macular degeneration and accurate information with respect to the known benefits of available treatments will impart a better understanding of this disease to patients.

Vascular endothelial growth factor expression and secretion by retinal pigment epithelial cells in high glucose and hypoxia is protein kinase C-dependent.

Retinal pigment epithelial (RPE) cells express vascular endothelial growth factor (VEGF) in response to high glucose or hypoxia. We hypothesised that VEGF expression and secretion by RPE cells in high glucose and hypoxia are regulated by protein kinase C (PKC). Primary cultured RPE cells from Sprague-Dawley rats were growth-arrested for 48 hr in 0.5% FBS in 5.6 or 30 mm D-glucose. Cells were exposed to hypoxic conditions (<1% O(2), 5% CO(2)) for the last 15-18 hr of growth-arrest. PKC -alpha, -beta(1), -delta, -epsilon, and -zeta were expressed by RPE cells and exposure to high glucose for 48 hr had no effect on expression as demonstrated by Western immunoblotting. High glucose, hypoxia or VEGF stimulated translocation of a number of the PKC isozymes to the membrane or particulate fractions implying activation. In response to high glucose or acute phorbol myristate acetate (PMA) stimulation, VEGF mRNA analysed by RT-PCR was increased. Intracellular VEGF protein identified by immunoblotting and confocal immunofluorescence imaging was significantly increased by high glucose, hypoxia or acute PMA stimulation. Calphostin C or a specific inhibitor of PKC-zeta prevented high glucose-stimulated VEGF expression in high glucose. VEGF secretion, as measured by ELISA in the culture medium, was enhanced in hypoxia but not in high glucose. Following exposure of RPE cells to PMA for 24 hr, PKC-delta was significantly down regulated, whereas PKC-alpha, -beta, -epsilon and -zeta remained unchanged. Secretion of VEGF in normal or high glucose, or hypoxia was significantly reduced following treatment with PMA for 24 hr but not with the PKC-zeta inhibitor. We conclude that in high glucose and hypoxia PKC isozymes are activated and are necessary for VEGF expression. Secretion of VEGF is enhanced in hypoxia and appears to be regulated by PKC-delta. RPE cells may contribute to the pathogenesis of retinopathy caused by high glucose and hypoxia through the expression and secretion of VEGF that are regulated by PKC isozymes.

Clinical assessment of conjunctival and episcleral vessel tortuosity in juvenile dermatomyositis.

PURPOSE: Conjunctival and episcleral vessel tortuosity are thought to be associated features of juvenile dermatomyositis. This study was conducted to determine the interobserver reliability of assessing normal and abnormal conjunctival and episcleral vessels from photographs of patients with and without juvenile dermatomyositis. METHODS: Color and red-free external ocular photographs of each eye of 28 children were assessed by 5 pediatric ophthalmologists from The Hospital for Sick Children. Fifteen of these pairs of photographs were of normal control eyes and 13 were from eyes of patients with juvenile dermatomyositis. On a standardized form each ophthalmologist was asked to determine whether both the conjunctival and episcleral vessels appeared normal or abnormal. Interobserver agreement was calculated by the weighted kappa statistic. The sensitivity and specificity of abnormal vessels for diagnosing juvenile dermatomyositis were also determined. RESULTS: Interobserver agreement for assessing conjunctival and episcleral vessels was 0.18 and -0.005, respectively. The average sensitivity and specificity of the evaluators for correctly assessing conjunctival vessels were 0.70 and 0.70, and for episcleral vessels, 0.67 and 0.47, respectively. CONCLUSIONS: There was low interobserver agreement in distinguishing between normal and abnormal eyes based on conjunctival and episcleral vessels. The sensitivity and specificity for identifying patients with juvenile dermatomyositis based on the appearance of vessels alone were relatively low. The appearance of conjunctival and episcleral vessels should be assessed in the context of other ocular and periocular findings of patients with juvenile dermatomyositis.