Quantitative analysis of posterior capsule opacification of hydrophobic acrylic intraocular lenses.

Posterior capsule opacification (PCO) remains a common complication of modern cataract surgery, although both modification of materials used and changes in the intraocular lens (IOL) optic edge design have helped to decrease its incidence slightly. Recently, various kinds of quantitative methods have been developed for measuring PCO. The purpose of this study was to compare the quantitative analysis of PCO between different types of IOL designs. Patients enrolled in the study had age-related cataract and underwent uneventful cataract surgery and implantation of either the AcrySof MA30BA (Alcon) or the Sensor AR40e (AMO), which are differently designed hydrophobic acrylic IOLs with a sharp-edged optic design. Postoperative examination was performed at 6 months. Retroillumination photographs of each eye were obtained, and the degree of PCO was assessed using the Evaluation of Posterior Capsule Opacification (EPCO) system. Grade 1 PCO was noted in both the MA30BA and the AR40e groups. There was no significant difference in the mean PCO score between the MA30BA and AR40e groups. Although the sharp-edged optic designs of both IOLs might similarly inhibit PCO at 6 months, a long-term follow-up period is needed to determine if any PCO differences occur between these 2 hydrophobic acrylic IOLs.

Two cases of true exfoliation of the lens capsule after cataract surgery.

True exfoliation of the lens capsule is known to be associated with glassblower's cataract, which is caused by extended exposure to excessive heat. Furthermore, inflammation and trauma are also considered to be predisposing factors. We report two cases of true exfoliation that were confirmed after cataract surgery. Neither patient exhibited true exfoliation before cataract surgery. In addition, neither patient had a history of occupation with exposure to excessive heat, inflammation or trauma. We observed the anterior lens capsules of these two patients with slit-lamp microscopy before and after cataract surgery. True exfoliation disappeared by adhering to the anterior capsule in both cases, and there were no complications during the observation period.

Inhibitory effects of Arg-Gly-Asp (RGD) peptide on cell attachment and migration in a human lens epithelial cell line.

Posterior capsule opacification (PCO) after cataract surgery is caused by growth of residual human lens epithelial (HLE) cells on the posterior capsule. We have shown that extracellular matrix (ECM) is an essential factor for HLE cell attachment and migration. The purpose of this study was to examine the inhibitory effects of Arg-Gly-Asp (RGD) peptide on cell attachment and migration in an HLE cell line. HLE cell line cells (SRA 01/04) that were obtained by transfection of large T antigen of SV40 were cultured in the absence of serum. The culture dishes were coated with type IV collagen, laminin or fibronectin, and Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) RGD peptide (0.1, 0.3, 1.0, 2.0 mg/ml) was added to the medium. The number of attached cells was counted after 90 min of incubation, and the inhibitory effects of GRGDSP RGD peptide on cell attachment were calculated. Cell attachment on the fibronectin-coated dishes was inhibited by GRGDSP RGD peptide at concentrations higher than 0.3 mg/ml; the inhibitory rate was 80% at a concentration of 2.0 mg/ml. The inhibition of cell attachment by GRGDSP RGD peptide on laminin-coated dishes appeared only at a concentration of 2.0 mg/ml, whereas no effects were observed on the type IV collagen-coated dishes. The inhibitory effects of GRGDSP RGD peptide on cell migration were measured in medium containing 2.0 mg/ml of GRGDSP RGD peptide after 1, 3, 5 and 7 days of culture. Cell migration was inhibited by GRGDSP RGD peptide from 1 day of culture on the fibronectin-coated dishes and from 5 days of culture on the laminin-coated dishes, whereas no effects were observed on the type IV collagen-coated dishes. GRGDSP RGD peptide inhibited cell attachment and migration on laminin and fibronectin that have RGD sequences. These data suggested that RGD peptide may have the potential to prevent PCO.

Idiopathic opacification of Berger's space.

We report a case of idiopathic opacification of Berger's space in a 68-year-old man. The opacification was in the retrolental space between the crystalline lens and the anterior vitreous in the right eye. Opaque fluid was surgically removed, and chemical analysis detected a high concentration of protein and a low concentration of mucopolysaccharides. No underlying pathology was observed.

Anterior chamber irrigation with an ozonated solution as prophylaxis against infectious endophthalmitis.

PURPOSE: To assess the validity of anterior chamber irrigation with an ozonated solution as prophylaxis against endophthalmitis. SETTING: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan. METHODS: Viability of human corneal endothelium in culture was assessed by the WST-8 assay, lactate dehydrogenase (LDH) release assay, and trypan blue exclusion assay after exposure to a 4 to 40 parts per million (ppm) solution for 10 to 60 seconds. The in vivo effect was observed 1 week after irrigation of a 4 ppm solution in the rabbit anterior chamber by trypan blue exclusion assay. Bactericidal efficacy of the anterior chamber irrigation with the 4 ppm solution was examined by bacterial colony count of the aqueous humor following methicillin-resistant Staphylococcus aureus (MRSA) contaminated intraocular lens implantation in the porcine eye. RESULTS: The WST-8 assay revealed no significant reduction of viability with 10-second exposure to a 4 ppm solution. Lactate dehydrogenase release and trypan blue exclusion assays similarly demonstrated little damage after 60-second exposure to a 4 ppm solution. In the rabbit cornea 1 week after treatment, damage caused by 30-second exposure to a 4 ppm solution was not significant. The MRSA colony count documented almost complete bactericidal action with 5-second exposure to the 4 ppm solution when no ophthalmic viscosurgical device existed in the anterior chamber. CONCLUSION: Limited damage to the corneal endothelium after 10-second exposure and potent bactericidal action with 5-second exposure suggests the validity of anterior chamber irrigation with a 4 ppm ozonated solution as prophylaxis against endophthalmitis.

[Diagnosis of ocular sarcoidosis by diagnostic criteria for systemic sarcoidosis].

PURPOSE: To evaluate the diagnostic criteria for systemic sarcoidosis in diagnosis of ocular sarcoidosis. SUBJECTS AND METHODS: Subjects were 105 ocular sarcoidosis suspects and 37 patients with other uveitis. We diagnosed ocular sarcoidosis suspects using the diagnostic criteria for systemic sarcoidosis proposed by the Japanese Committee for Diffuse Lung Diseases. The criteria included histological and clinical diagnosis, and the clinical diagnosis required 5 systemic tests: 1) tuberculin skin test, 2) serum gamma-globulin(gamma-gl), 3) serum angiotensin converting enzyme(ACE), 4) serum lysozyme, and 5) 67Ga scintigraphy. Three positive findings including either 1) or 3) fulfilled the clinical diagnosis. RESULTS: Sixty-two patients were histologically and/or clinically diagnosed, and 43 patients remained undiagnosed. The histological and clinical diagnosis did not produce the same diagnostic yields. The sensitivity of ACE and gamma-gl was low. The percentage of patients showing increased lymphocytosis and/or CD 4/8 in bronchoalveolar lavage was similarly high in the diagnosed and undiagnosed, suggesting the presence of definitive ocular sarcoidosis in the undiagnosed. CONCLUSIONS: The diagnostic criteria for systemic sarcoidosis yielded false negative results in diagnosing ocular sarcoidosis. The selection and combination of systemic tests for clinical diagnosis should be further studied.

Polyriboinosinic polyribocytidylic acid [poly(I:C)]/TLR3 signaling allows class I processing of exogenous protein and induction of HIV-specific CD8+ cytotoxic T lymphocytes.

In the case of viral infection, various viral proteins and genetic components are disseminated in the body. The former viral proteins may be captured by immature dendritic cells (DC) and the latter genetic components may stimulate the antigen-loading DC to maturate via specific Toll-like receptors (TLR), leading to the establishment of virus-specific cellular immunity; in particular, cytotoxic T lymphocytes (CTL) that control intracellular virions. Polyriboinosinic polyribocytidylic acid [poly(I:C)], which might reflect a natural genetic product from a variety of viruses during replication, has recently been identified as one of the critical stimuli for TLR3. Based on these observations, we speculated that stimulation of TLR3 with poly(I:C) might drive the direction of acquired/adaptive immunity to the cellular arm. Indeed, when BALB/c mice were immunized with purified recombinant HIV-1 envelope gp120 or influenza hemagglutinin (HA) protein together with poly(I:C), epitope-specific CD8(+) class I MHC molecule-restricted CTL were primed from naive CD8(+) T cells in vivo. In contrast, when the same proteins were immunized with lipopolysaccharide, a stimulant of TLR4, specific CTL were not primed at all. Moreover, we show here that immature DC could present processed antigen from captured purified protein in association with class I MHC molecules in the presence of poly(I:C), but not of LPS. These results indicate that we are able to manipulate the direction of acquired/adaptive effector immune responses using an appropriate stimuli and the findings presented in this paper will offer a new therapeutic strategy using poly(I:C) administration for priming antigen-specific CD8(+) CTL with purified viral protein in vivo.

Lentivirus-mediated expression of angiostatin efficiently inhibits neovascularization in a murine proliferative retinopathy model.

Ischemic retinal diseases, such as diabetic retinopathy, retinopathy of prematurity, and age-related macular degeneration, are a major cause of blindness worldwide. Angiostatin is an internal peptide fragment of plasminogen that inhibits endothelial proliferation in vitro and tumor growth in vivo. We now demonstrate that HIV vector encoding angiostatin (HIV-angiostatin) can inhibit retinal neovascularization in a mouse model of proliferative retinopathy. Intravitreal injections of HIV-angiostatin led to stable expression of the angiostatin gene in retinal tissue. Retinal neovascularization was histologically quantitated by a masked protocol. Retinal neovascularization in the eye injected with HIV-angiostatin was reduced in 90% (9/10; P=0.025) of animals, compared with the eye injected with phosphate-buffered saline. Reduction of histologically evident neovascular nuclei per 6-microm section averaged 68%, with maximal inhibitory effects of 87%. Neovascularization was not reduced in the eyes injected with HIV vector encoding enhanced green fluorescent protein. This is the first report that HIV-angiostatin can reduce neovascular cell nuclei in a murine proliferative retinopathy model. These data suggest that the anti-angiogenic activity of angiostatin has therapeutic potential for the treatment of retinal neovascularization.

Free radicals in phacoemulsification and aspiration procedures.

OBJECTIVES: To detect free radicals in phacoemulsification and aspiration procedures using electron-spin resonance and to observe the effect of ophthalmic viscosurgical devices (viscoelastic agents) on free radical intensity. METHODS: (1) A test tube containing BSS Plus (Alcon Laboratories, Inc, Fort Worth, Tex) with 1% of the spin-trapping agent, 5,5'-dimethyl-1-pyrroline N-oxide, without irrigation and aspiration, was exposed to ultrasound (100% for 20 seconds). A preparation of hyaluronate sodium (Healon [a cohesive agent that contains 1% hyaluronate sodium] (Pharmacia, Uppsala, Sweden) or Viscoat [a dispersive agent that contains 3% hyaluronate sodium and 4% chondroitin sulfate] (Alcon Laboratories, Inc)) was added to the solution to observe inhibitory effects. (2) To simulate a clinical procedure, an eye model with irrigation and aspiration of a combination of 1% 5,5'-dimethyl-1-pyrroline N-oxide and BSS Plus, 25 mL/min, as the irrigating solution was exposed to ultrasound (for 10, 20, or 30 seconds). Healon or Viscoat was injected into the anterior chamber. Free radicals were measured by an electron-spin resonance spectrometer. RESULTS: (1) A characteristic signal corresponding to hydroxyl radicals was detected. Similar inhibition by Healon and Viscoat was observed. (2) Two ophthalmic viscosurgical devices similarly suppressed the signal at 10 seconds. The inhibition by Healon ceased at 20 seconds, whereas Viscoat suppressed the signal throughout the time course. CONCLUSIONS: Phacoemulsification produces hydroxyl radicals in the anterior chamber even with irrigation and aspiration. The effect of ophthalmic viscosurgical devices on free radicals depends on the retention of the materials within the anterior chamber. CLINICAL RELEVANCE: There are complications associated with phacoemulsification.

[Ocular sarcoidosis].

Ocular involvement of sarcoidosis is frequent, and it is often the initial clinical manifestation of the disease. The most common ocular lesions include granulomatous uveitis associated with iris and trabecular nodules, string of pearl-type vitreous opacities, retinal paerivasculitis mainly affecting veins, and patchy retino-choroidal exudates. A half of the patients with typical ocular lesions suggestive of sarcoidosis did not show the systemic evidence, and they remained as sarcoidosis suspects. Risks of visual deterioration are secondary glaucoma, vitreous opacities, cystoid macular edema, and retinal neovascularization. Thirty-four % of the patients were treated with systemic corticosteroids, and some patients required other treatment such as methotraxate. Twenty-one% of the patients resulted in the poor visual acuity of less that 0.5.

New strategy for in vivo transgene expression in corneal epithelial progenitor cells.

PURPOSE: Efficient in vivo gene transfer into corneal epithelial cells and stable transgene expression would have broad clinical applications. We therefore assessed the capacity of adenoviral, adeno-associated viral (AAV) and lentiviral vectors encoding enhanced green fluorescent protein (EGFP) to transduce rat corneal epithelial progenitor cells. METHODS: Superficial cells of the corneal epithelium were shaved, after which 20 microl of the respective vector solutions were inoculated onto the basal cell layer of the limbal epithelium for 30 min. Results. Three days after transduction, fluorescence microscopic examination revealed the presence of EGFP+ cells in all corneas, irrespective of the vector used; however, EGFP+ cells were undetectable in corneas transduced with adenoviral and AAV vectors after 2 and 4 weeks, respectively. By contrast, EGFP+ cells were still detected among cells transduced with lentiviral vector 6 weeks after transduction. DAPI (4', 6-diamidino-2-phenylindole) staining confirmed that it was the basal layer cells that continued to express EGFP throughout the 6-week period. CONCLUSIONS: The use of a lentiviral vector for in vivo transfer of foreign genes into corneal epithelial stem and transient amplifying cells may represent a new and effective approach to the treatment of corneal disease, as well as to the study of the biology of these stem cells.