Vascular endothelial growth factor-induced retinal permeability is mediated by protein kinase C in vivo and suppressed by an orally effective beta-isoform-selective inhibitor.

Increased vascular permeability and excessive neovascularization are the hallmarks of endothelial dysfunction, which can lead to diabetic macular edema and proliferative diabetic retinopathy in the eye. Vascular endothelial growth factor (VEGF) is an important mediator of ocular neovascularization and a known vasopermeability factor in nonocular tissues. In these studies, we demonstrate that intravitreal injection of VEGF rapidly activates protein kinase C (PKC) in the retina at concentrations observed clinically, inducing membrane translocation of PKC isoforms alpha, betaII, and delta and >threefold increases in retinal vasopermeability in vivo. The effect of VEGF on retinal vascular permeability appears to be mediated predominantly by the beta-isoform of PKC with >95% inhibition of VEGF-induced permeability by intravitreal or oral administration of a PKC beta-isoform-selective inhibitor that did not inhibit histamine-mediated effects. These studies represent the first direct demonstration that VEGF can increase intraocular vascular permeability through activation of PKC in vivo and suggest that oral pharmacological therapies involving PKC beta-isoform-selective inhibitors may prove efficacious for the treatment of VEGF-associated ocular disorders such as diabetic retinopathy.

Progress in measurement of ocular blood flow and relevance to our understanding of glaucoma and age-related macular degeneration.

New technologies have facilitated the study of the ocular circulation. These modalities and analysis techniques facilitate very precise and comprehensive study of retinal, choroidal, and retrobulbar circulations. These techniques include: 1. Vessel caliber assessment; 2. Scanning laser ophthalmoscopic fluorescein angiography and indocyanine green angiography to image and evaluate the retinal circulation and choroidal circulation respectively; 3. Laser Doppler flowmetry and confocal scanning laser Doppler flowmetry to measure blood flow in the optic nerve head and retinal capillary beds; 4. Ocular pulse measurement; and 5. color Doppler imaging to measure blood flow velocities in the central retinal artery, the ciliary arteries and the ophthalmic artery. These technique have greatly enhanced the ability to quantify ocular perfusion defects in many disorders, including glaucoma and age-related macular degeneration, two of the most prevalent causes of blindness in the industrialized world. Recently it has become clear, in animal models of glaucoma, that retinal ganglion cells die via apoptosis. The factors that initiate apoptosis in these cells remain obscure, but ischemia may play a central role. Patients with either primary open-angle glaucoma or normal-tension glaucoma experience various ocular blood flow deficits. With regard to age-related macular degeneration, the etiology remains unknown although some theories include primary retinal pigment epithelial senescence, genetic defects such as those found in the ABCR gene which is also defective in Stargardt's disease and ocular perfusion abnormalities. As the choriocapillaris supplies the metabolic needs of the retinal pigment epithelium and the outer retina, perfusion defect in the choriocapillaris could account for some of the physiologic and pathologic changes in AMD. Vascular defects have been identified in both nonexudative and exudative AMD patients using new technologies. This paper is a comprehensive update describing modalities available for the measurement of all new ocular blood flow in human and the clinical use.

Photodynamic therapy with PhotoPoint photosensitiser MV6401, indium chloride methyl pyropheophorbide, achieves selective closure of rat corneal neovascularisation and rabbit choriocapillaris.

AIM: The new photosensitiser PhotoPoint MV6401, indium chloride methyl pyropheophorbide, was assessed as a possible ocular photodynamic therapy agent in a rat model of experimentally induced corneal neovascularisation and in choriocapillaris closure in the rabbit. Optimal drug and light activation parameters were determined. METHODS: MV6401 (Miravant Pharmaceuticals, Inc, Santa Barbara, CA, USA) was activated at 664 nm using a DD3-0665 (Miravant Systems Inc) 0.5 W diode laser. Corneal neovascularisation in rats was induced using an N-heptanol technique. The evaluated drug dosages, light dosages, and post-injection activation times ranged from 0.01-0.1 micromol/kg, 5-25 J/cm(2), and 10-60 minutes, respectively. The efficacy of MV6401 on normal choriocapillaris and choroidal vessels was evaluated in rabbits with indirect ophthalmoscopy, fundus photography, fluorescein angiography, and histology. In rabbits, the evaluated drug dosages, light dosages, and post-injection activation times ranged from 0.025-0.25 micromol/kg, 3.3-20 J/cm(2), and 10 minutes, respectively. RESULTS: In the rat corneal neovascularisation model, an optimal intravenous drug dosage of 0.075 micromol/kg was activated by a 20 J/cm(2) light dose at 10 minutes after drug administration, the results of which demonstrated early evidence of efficacy in ocular neovascularisation. In rabbits, closure of the normal choriocapillaris was selectively achieved at a drug dosage of 0.15 micromol/kg using light doses from 3.3 to 20 J/cm(2). CONCLUSION: PhotoPoint MV6401 is a potent photosensitiser that demonstrates both efficacy and selectivity in experimental ocular models.

Relation between dietary intake, serum concentrations, and retinal concentrations of lutein and zeaxanthin in adults in a Midwest population.

BACKGROUND: Information on concentrations of retinal carotenoids (macular pigment, or MP) is of particular interest because MP protects against age-related macular degeneration, the leading cause of irreversible blindness in the United States. OBJECTIVE: This study was designed to evaluate the relation between dietary intake, blood concentrations, and retinal concentrations of carotenoids in a large group of volunteers. DESIGN: Two hundred eighty volunteers in the Indianapolis area completed health and diet questionnaires, donated a blood sample, and participated in MP density assessment to determine retinal carotenoid status. Dietary intake was assessed by food-frequency questionnaire. Serum concentrations of lutein, zeaxanthin, and beta-carotene were measured by HPLC. MP optical density (MPOD) was determined psychophysically with a 460-nm, 1 degrees test stimulus. RESULTS: Average MPOD was 0.21 +/- 0.13. Average intakes of lutein + zeaxanthin and beta-carotene were 1101 +/- 838 and 2935 +/- 2698 microg/d, respectively. Although several key dietary intake variables (eg, lutein + zeaxanthin and beta-carotene) differed by sex, no significant sex differences were found in either serum concentrations of lutein and zeaxanthin or MPOD. Serum beta-carotene concentrations were significantly higher in women than in men. Serum lutein + zeaxanthin and dietary intake of lutein + zeaxanthin were significantly correlated and significantly related to variations in MPOD (r = 0.21, P < 0.001, and r = 0.25, P < 0.001, respectively). CONCLUSIONS: Retinal carotenoids can be measured in epidemiologic studies. In this study, MPOD was associated with lutein + zeaxanthin in the diet and the serum. Retinal concentrations, however, were influenced by other factors as well. To understand the effect of dietary lutein + zeaxanthin intake on the retina and risk of age-related eye disease, future studies should include measures of macular concentrations of these pigments.

Macular pigment optical density before and after cataract extraction.

PURPOSE: Psychophysical methods of measuring macular pigment (MP) use comparisons of short- and midwave light in the fovea and parafovea to derive optical density estimates. This light must pass through the crystalline lens before absorption by the MPs can occur. The effect of lens absorption on these measures has not been adequately determined. The present study assesses the influence of lens absorption on MP measurements by comparing MP optical density (MPOD) measured before and after cataract extraction. METHODS: MPOD was measured using flicker photometry in free view at 458 nm with a 1 degrees stimulus. Twenty-nine eyes from 24 patients with cataracts sufficiently severe to require cataract extraction were evaluated. RESULTS: In the entire group of 24 patients, the mean (+/-SD) age measured 68.7 +/- 9.5 years, and the mean MPOD measured 0.19 +/- 0.11. For all 29 eyes measured, MPOD averaged 0.206 +/- 0.13 before and 0.18 +/- 0.12 after cataract extraction. MPOD measurements at the two time points (mean 8.1 +/- 4.7 weeks after surgery) were highly correlated (r = +0.58), suggesting that a cataractous lens does not influence the MP measurement technique. CONCLUSIONS: Psychophysical techniques can be used to obtain reliable measurements of MP in elderly subjects, even in those with cataracts. Moreover, differences in retinal illuminance due to varying opaqueness of the crystalline lens do not seem to have a measurable influence on MPOD.

Transpupillary thermotherapy for subfoveal occult choroidal neovascularization: effect on ocular perfusion.

PURPOSE: To perform a descriptive analysis of the effects on ocular blood flow of transpupillary thermotherapy (TTT) for occult subfoveal choroidal neovascular membranes (CNVMs) in age-related macular degeneration (AMD). METHODS: Eleven subjects with occult subfoveal CNVM due to AMD were assessed in a masked fashion by color Doppler imaging (CDI) within 24 hours before, 24 hours after, and 1 month after undergoing TTT. RESULTS: In the posterior ciliary arteries (PCAs), there were no statistically significant changes observed in the peak systolic velocity (PSV), end diastolic velocity (EDV), or resistive index (RI) at 24 hours. At 1 month, the mean EDV decreased 36% (P = 0.0105) and the mean RI increased 3.8% (P = 0.0305) in the nasal PCA. Although there was a similar trend in the temporal PCA, the differences did not reach statistical significance. In the central retinal artery (CRA), the mean PSV decreased 16% (P = 0.0137), and the mean EDV decreased 21% (P = 0.0222) at 24 hours after treatment. There were no statistically significant differences in the CRA blood flow indices at 1 month after treatment. In the ophthalmic artery, there were no statistically significant differences observed in the mean PSV, EDV, or RI at 24 hours or 1 month after treatment. CONCLUSIONS: TTT is associated with transiently decreased volumetric blood flow in the retinal circulation 24 hours after treatment. In the posterior ciliary arteries that supply the choroid, there were no changes observed at 24 hours, but at 1 month, there was a decrease in the mean EDV and an increase in the RI in the nasal and temporal PCAs, reaching statistical significance in the nasal PCA only. This study suggests that TTT could lead to alterations in choroidal blood flow, as assessed by CDI. Further study is warranted.

Regulation of retinal and optic nerve blood flow.

Blood flow to the retina and optic nerve remains constant over a range of elevated intraocular pressure or mean arterial pressure, independent of sympathetic activation (pressure autoregulation). In addition, increased metabolic activity in these tissues proportionally increases blood flow (metabolic autoregulation). At constant metabolic rate, altered arterial oxygen content reciprocally alters blood flow, leaving total oxygen delivery constant, while blood flow rises and falls with the arterial carbon dioxide tension. These responses are similar to those of the cerebral circulation. However, while aging, atherosclerosis, arterial hypotension, and individual variation may profoundly alter blood flow regulation and predispose to the development of illness, these factors remain largely unexplored.

Intravitreal triamcinolone acetonide inhibits choroidal neovascularization in a laser-treated rat model.

OBJECTIVE: To determine if intravitreal triamcinolone acetonide (TAAC) inhibits experimental choroidal neovascular membranes induced by laser trauma in a rat model. METHODS: Nineteen anesthetized male Brown Norway rats received a series of 8 krypton red laser lesions per eye (647 nm, 0.05 seconds, 50 microm, and 150 mW in 17 rats, and 200 mW in 2 rats). One eye received an intravitreal injection of triamcinolone acetonide (20 microL, 0.8 mg) and the other eye received an injection of isotonic sodium chloride solution. Fundus and fluorescein angiography examinations occurred just before euthanasia and tissue processing for histopathology on day(s) 0, 1, 3, 7, 14, 21, 28, and 35. RESULTS: From the control eyes that underwent photocoagulation at 150 mW, 57 discrete lesions with definitive fibrovascular proliferations were observed at 21, 28, and 35 days, arising from a total of 72 spots placed (79% yield). From the control eyes that underwent photocoagulation at 200 mW, 11 discrete lesions with definitive fibrovascular proliferations were observed at 28 days, arising from a total of 16 spots placed (69% yield). In the TAAC-treated group, no fibrovascular proliferations were observed in the 72 lesions and in the 16 lesions created with 150 mW and 200 mW, respectively. CONCLUSION: Intravitreal TAAC is a potent inhibitor of fibrovascular proliferations in a rat model of choroidal neovascular membranes induced by laser trauma. CLINICAL RELEVANCE: This study corroborates previous investigations that propose TAAC as a potential treatment for choroidal neovascular membranes in humans.

Aging affects the retrobulbar circulation differently in women and men.

BACKGROUND: While aging clearly has protean biological effects on every organ system, the differential effects of aging in women and men in the retrobulbar vasculature, to our knowledge, have never been investigated. Because glaucoma and age-related macular degeneration are closely linked to advanced age, we performed a cross-sectional study using color Doppler imaging of 4 retrobulbar vessels in both healthy women and men. OBJECTIVE: To define the influence of aging per se on ocular hemodynamics. METHODS: Women (n = 73) and men (n = 55), aged from 20 to 90 years, free of ocular and systemic disease, and with normal intraocular pressure, were recruited for this study. Postmenopausal women who were not receiving estrogen replacement therapy were also recruited. Studies involved color Doppler imaging analysis of the ophthalmic, central retinal, and nasal and temporal posterior ciliary arteries. Ophthalmic arterial peak systolic and end-diastolic velocities and a Pourcelot resistance index were determined for each vessel. RESULTS: In both sexes, ophthalmic arterial end-diastolic velocity decreased and the Pourcelot resistance index rose with advancing age (each P<. 001); peak systolic velocity in the ophthalmic vessel was age-independent. In contrast, central retinal arterial flow velocities were unaffected by age in both sexes. In the posterior ciliary arteries, in men, flow velocities and the Pourcelot resistance index were independent of age. However, in women, end-diastolic velocity decreased with age in both the nasal and temporal posterior ciliary vessel (each P<.05); peak systolic velocity was constant; the Pourcelot resistance index in each ciliary artery rose with advancing age (each P<.05). CONCLUSION: In healthy women and men, aging-induced changes in retrobulbar hemodynamics are comparable to alterations seen in patients with glaucoma or age-related macular degeneration, suggesting that vascular changes with senescence may contribute to increased risk for these diseases in older age. Arch Ophthalmol. 2000;118:1076-1080

Primary parathyroid hyperplasia associated with thyroid hemiagenesis and agenesis of the isthmus.

BACKGROUND: Congenital thyroid anomalies can be caused by abnormal descent or agenesis of part of the thyroid gland. Agenesis of the thyroid gland may involve one or both lobes of the gland, with or without isthmus, and is very rare. Congenital thyroid anomalies have been reported to be associated with thyroid diseases (hyperthyroidism, adenomatous goiter, and cancer) but not with parathyroid diseases. METHODS: Case reports of two patients describe the methods used. RESULTS: We report two patients with hyperparathyroidism and congenital thyroid anomalies. One patient had a left thyroid lobe hemiagenesis and the other patient had agenesis of the thyroid isthmus. Both patients had primary hyperparathyroidism caused by parathyroid hyperplasia and were treated by subtotal parathyroidectomy. The parathyroid glands and recurrent laryngeal nerves were in their usual positions despite the abnormal development of the thyroid glands. CONCLUSIONS: When thyroid hemiagenesis or isthmus agenesis is found in patients undergoing parathyroidectomy for hyperparathyroidism, parathyroid hyperplasia should be suspected. If parathyroid hyperplasia is present, subtotal parathyroidectomy and bilateral thymectomy are recommended. The parathyroid glands and the recurrent laryngeal nerves can be found in their usual locations.

Age-related macular degeneration: a review of experimental treatments.

Age-related macular degeneration (AMD) is the leading cause of irreversible visual loss in the USA. Laser photocoagulation of choroidal neovascular membranes (CNVMs) in exudative AMD is currently the only well-studied and widely accepted treatment modality. It is beneficial for only a small minority of patients who show well-demarcated "classic" CNVMs, and it destroys normal retinal tissue, creates a scotoma, and is associated with an unacceptably high CNVM persistence and recurrence rate. Consequently, investigators have attempted to develop new modalities for treatment of CNVMs. These treatment modalities can be grouped into four major categories: photodynamic therapy; pharmacologic inhibition of CNVM formation with antiangiogenic agents; surgical intervention, including excision of subfoveal CNVMs; and radiation therapy. All of these experimental treatment modalities are directed toward destroyiing CNVMs, the end result of the exudative process, and all have limitations. The ideal treatment of the future must be based on the pathogenesis of the disease at a stage well before CNVMs develop. Investigations in nonexudative AMD are currently focusing on several major areas. Epidemiologic factors, such as genetics, sunlight, and nutrition, are being evaluated in several large studies, including the Age-Related Eye Disease Study, with the possibility of ultimately limiting the risk of AMD through behavior modification. Laser treatment of drusen is being evaluated as a means of limiting the risk of CNVM formation, although mixed results have been reported in the small number of studies to date. Choroidal perfusion abnormalities have been described in AMD, and some investigators postulate that altering blood flow may limit the risk of CNVM formation. No perfusion-treatment trials have been completed to date.

Vitreous amyloidosis in familial amyloidotic polyneuropathy. Report of a case with the Val30Met transthyretin mutation.

We present a clinical pathological review of vitreous amyloidosis in a case of familial amyloidotic polyneuropathy, type I. Vitreous opacification was the first manifestation of disease in the proband, who was successfully treated with vitrectomy. The eyes were obtained at autopsy after the patient died from an unrelated cause, and the histopathology is presented here. Analysis of DNA from the pathology specimen revealed the most commonly reported transthyretin mutation, Val30Met. The classification of systemic and ocular amyloidosis as well as the genetics of familial amyloidotic polyneuropathy are briefly reviewed.

Uveal lymphoid neoplasia: a clinical-pathologic correlation and review of the early form.

We report three cases of uveal lymphoid neoplasia that were diagnosed early in their course. One case exhibited a posterior form, presenting with progressive hyperopia from a serous-macular detachment and choroidal involvement along with retrobulbar involvement. This patient was treated with proton beam irradiation. Two cases displayed an anterior form, with fixed fleshy epibulbar masses resembling salmon patches, and choroidal involvement. The histologic findings from biopsy of these anterior masses are presented. One of these patients was treated with complete excision of the mass and double freeze-thaw cryotherapy of the scleral bed, and the other patient was treated with conventional photon beam irradiation. The clinical features of uveal lymphoid neoplasia in its early form are discussed. Evaluation and treatment strategies for these early forms of uveal lymphoid neoplasia are reviewed.

Acute progressive multifocal Best's disease in a 61-year-old man.

PURPOSE: To report acute progressive multifocal Best's disease in patients in their seventh decade. METHOD: We report one such case. RESULTS: We examined a 61-year-old man with strikingly symmetric, 1-disk diameter serous retinal detachments involving each fovea. Fluorescein angiography was unremarkable, with no dye leakage. Electrophysiologic testing confirmed the diagnosis. Visual acuity decreased, and within 1 month, the size and number of lesions rapidly increased. Three months later, the lesions quickly evolved to the classic vitelliform stage. CONCLUSION: This well-documented case expands the spectrum of findings in multifocal Best's disease.

Talc embolism: a static retinopathy.

PURPOSE: To document the static nature of crystalline deposits in talc retinopathy. METHOD: Case report. RESULTS: A 38-year-old woman with a remote history of intravenous drug abuse showed the incidental finding of fine, irregularly shaped refractile deposits in the retinal microvasculature. At 2-year follow-up, fundus findings were identical to the initial manifestation with respect to the distribution and position of crystalline deposits in each fundus. CONCLUSION: This case documents the static nature of crystalline deposits in talc retinopathy.

Subretinal aspiration biopsy of ocular lymphoma.

PURPOSE: Large-cell lymphoma can be difficult to diagnose because vitreous biopsies may fail to disclose neoplastic cells. METHODS: We report one such case in which diagnosis was confirmed by subretinal aspiration of yellow-white infiltrates using a pars plana approach. RESULTS: A 67-year-old woman with recurrent bilateral uveitis was diagnosed with large-cell lymphoma by subretinal aspiration of yellow-white infiltrates after two vitreous biopsies and a concurrent retinal biopsy failed to confirm the diagnosis. CONCLUSIONS: When the suspicion of intraocular lymphoma remains high despite previous negative vitreous biopsies, retinal biopsy and aspiration biopsy of subretinal lesions may enhance the diagnostic yield.

Postvitrectomy positioning complicated by ulnar nerve palsy.

PURPOSE: Ulnar nerve palsy is an unusual complication of postvitrectomy positioning. METHOD: We report two such cases. RESULTS: Both patients developed ulnar nerve palsy after maintaining a face-down position for 2 to 4 weeks after vitrectomy with intraocular perfluoro-octane. The neuropathy resolved in one patient after he was instructed to avoid prolonged pressure on his flexed elbows, whereas the other patient ultimately required surgical decompression of the nerve at the elbow. CONCLUSION: Ulnar nerve palsy can result from postvitrectomy positioning associated with prolonged direct pressure on the ulnar nerve or compression of the ulnar nerve via flexion of the elbow.

Evolution of a full-thickness macular hole.

PURPOSE: To illustrate succinctly the various stages of macular hole formation. METHOD: We photodocumented the evolution of a full-thickness macular hole in a patient who refused surgery. RESULTS: A 60-year-old woman, who had had difficulty reading for 3 months, was initially examined with a stage 1B macular hole in her right eye that progressed to stage 2 over a 6-month period and then to stage 3, 2 years after initial examination. CONCLUSIONS: This case demonstrates vividly and succinctly the development of a full-thickness macular hole from stage 1B to stage 3 and provides a unique opportunity to study carefully in a single eye the sequential progression of this important retinal disorder.

Mitochondrial maculopathy: geographic atrophy of the macula in the MELAS associated A to G 3243 mitochondrial DNA point mutation.

PURPOSE: To report ocular findings in the mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS syndrome) in a family with the A to G 3243 mitochondrial (mt) DNA point mutation. METHODS: Case reports. Ocular findings are described from four family members with the MELAS associated A to G 3243 mt DNA point mutation. RESULTS: Findings included ophthalmoplegia, neurosensory deafness, reduction of photopic and scotopic electroretinogram b-wave amplitudes, and myopathy, as well as macular retinal pigment epithelial atrophy. No family members had nyctalopia, attenuation of retinal blood vessels, or retinal bone spicule pigmentation. CONCLUSION: The finding of slowly progressive macular retinal pigment epithelial atrophy expands the reported phenotypic diversity of patients with A3243G mt DNA mutations.

Comparison of intraocular to plasma HIV-1 viral burden in patients with cytomegalovirus retinitis.

PURPOSE: To determine the relationship between plasma and intraocular human immunodeficiency virus-1 (HIV-1) viral loads in 12 consecutive patients undergoing ganciclovir implant surgery for cytomegalovirus (CMV) retinitis. METHODS: Aqueous and vitreous specimens were assayed for HIV-1 viral load by polymerase chain reaction analysis (Roche Amplicor HIV Monitor; Roche Diagnostics Systems, Inc, Branchburg, New Jersey). RESULTS: It was possible to quantitatively assay HIV-1 burden in intraocular fluids using polymerase chain reaction analysis. In general, patients with plasma viral loads less than 250,000 copies/ml had undetectable (<200 copies/ml) HIV-1 in their aqueous and vitreous. CONCLUSIONS: It is likely that intraocular viral levels have several determinants in addition to plasma viral loads, with which they only partially correlate.