Clinical significance of topical instillation technique in Japanese glaucoma patients.

The clinical significance of a proper eye drop application technique was evaluated in Japanese glaucoma patients. Patients diagnosed with primary open-angle glaucoma having intraocular pressure (IOP) greater than 21 mmHg were treated with eye drops at home. In some patients, however, the topical treatment was ineffective. They returned to the hospital to receive surgical treatment. On admission, 56% of these patients had IOP greater than 21 mmHg. Patient instillation technique was evaluated based on the proximity of the eyedropper tip to the eyes, application position, eyelid closure, treatment (removal) of excess fluid, and nasolacrimal occlusion. In addition, pharmacists interviewed patients to determine the level of understanding of glaucoma, knowledge of prescribed drugs, home application technique, and sensation after application. Multivariate analysis revealed that the key factors influencing the control of IOP to less than 21 mmHg with topical medication were: application of drops in the center of the eye and removal of excessive fluid, in addition to gender and age. Proper topical application at home was dependent on the patient's understanding of the disease, knowledge of prescribed drugs, patient education on the use of drugs, the competence of the instructor, and knowledge of correct application technique. This study indicates that easily comprehensible patient education on the use of eye drops, the nature of glaucoma and the proper use of prescribed drugs is vital to improving the clinical efficacy of topical ophthalmic medication of glaucoma in adult patients.

Aqueous humour levels of cytokines are correlated to vitreous levels and severity of macular oedema in branch retinal vein occlusion.

AIM: To investigate whether the aqueous levels of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) are correlated to the vitreous levels of these substances and to the severity of macular oedema in branch retinal vein occlusion (BRVO). METHODS: Aqueous and vitreous samples were obtained during cataract and vitreous surgery from 24 patients (24 eyes) with macular oedema in BRVO. The VEGF and IL-6 levels in aqueous humour, vitreous fluid, and plasma were determined by enzyme-linked immunosorbent assay. The degree of retinal ischaemia was evaluated in terms of the area of capillary nonperfusion using the Scion Image. The severity of macular oedema was evaluated using the OCT. RESULTS: The aqueous level of VEGF was significantly correlated with the vitreous level of VEGF (P<0.0001). Vitreous levels of VEGF and IL-6 were significantly correlated with the nonperfusion area of BRVO (P<0.0001, P=0.0061, respectively), as were the aqueous levels of VEGF and IL-6 (P<0.0001, P=0.0267, respectively). Furthermore, the vitreous levels of VEGF and IL-6 and the aqueous level of VEGF were significantly correlated with the severity of macular oedema of BRVO (P=0.0001, P=0.0331, P=0.0272, respectively). CONCLUSION: Our results suggest that the aqueous level of VEGF may reflect its vitreous level. Measurement of the aqueous level of VEGF may be clinically useful to indicate the severity of macular oedema with BRVO.

Subretinal neovascularization associated with retinochoroidal coloboma.

PURPOSE: To report a case of subretinal neovascularization associated with retinochoroidal coloboma. METHODS AND RESULTS: A 44-year-old female presented with metamorphopsia in her right eye for 4 weeks. Funduscopic examination revealed bilateral inferior retinochoroidal coloboma. Fluorescein angiography disclosed foci of subretinal neovascularization at the margin between the colobomatous defect and the normal-appearing retina. Five month later, multiple small areas of subretinal hemorrhages were noted. The hemorrhage was gradually absorbed. Six years after initial presentation, subretinal hemorrhage did not recur and her right VA was 0.2. CONCLUSIONS: Ophthalmologists should be aware of this rare but important complication of retinochoroidal coloboma.

Cataract in atomic bomb survivors.

PURPOSE: Ophthalmologic examinations were conducted on atomic bomb (A-bomb) survivors 55 years after exposure. MATERIALS AND METHODS: A-bomb survivors who had been exposed before 13 years of age at the time of the bombings in 1945 or who had been examined in a previous study between 1978 and 1980. The examinations, conducted between June 2000 and September 2002, included slit-lamp examination, digital photography and a cataract grading system for three parts of the lens (nucleus, cortex and posterior subcapsule) as an outcome variable. Proportional odds logistic regression analysis was conducted using the lowest grading class as a reference and included explanatory variables such as age, sex, city, dose and various cataract-related risk factors. When the grades in an individual differed, the worst grade was used. RESULTS: Results indicate that odds ratios (ORs) at 1 Sv were 1.07 (95% confidence intervals [CI] 0.90, 1.27) in nuclear colour, 1.12 (95% CI 0.94, 1.30) in nuclear cataract, 1.29 (95% CI 1.12, 1.49) in cortical cataract and 1.41 (95% CI 1.21, 1.64) in posterior subcapsular cataract. The same was true after excluding 13 people whose posterior subcapsular cataracts had been previously detected. CONCLUSION: Significant radiation effects were observed in two types of cataracts in A-bomb survivors.

Characteristics and surgical outcomes of paediatric retinal detachment.

PURPOSE: To report the clinical features and surgical and visual outcomes of rhegmatogenous retinal detachment (RRD) in the paediatric population. METHODS: A retrospective review of children (aged 0-15 years) who underwent primary surgical repair for RRD at the Hiroshima University Hospital between 1988 and 2001. RESULTS: In all 53 eyes of 49 patients were identified; paediatric RRD accounted for 3.1% of 1779 eyes with RRD operated on during this period. The causes of RRD included blunt trauma (27%), myopia (25%), idiopathic (20%), familial exudative vitreoretinopathy (13%), and others. Among 55 eyes, 12 (22%) already had proliferative vitreoretinopathy (PVR) of grade C or D preoperatively. The median initial visual acuity (VA) was 0.3. Retinal reattachment was achieved with a single operation in 78%. Final retinal reattachment was achieved in 87%. Retinal reattachment rates with and without PVR were 42% and 100%, respectively (P<0.01). Median final VA was 0.7. Final VA was > or =0.1 in 73% and > or =0.5 in 53%; four eyes had a final VA of no light perception. The presence of preoperative PVR (P=0.03) and the initial VA (P<0.0001) significantly affected final VA. CONCLUSIONS: Paediatric RRD is characterised by a delay in diagnosis, as evidenced by the high rate of PVR at presentation. Retinal reattachment was adversely affected by the presence of PVR. Final VA correlated with the initial VA and was significantly affected by preoperative PVR. Early diagnosis may improve the visual prognosis of paediatric retinal detachment.

Autologous blood injection for marked overfiltration early after trabeculectomy with mitomycin C.

PURPOSE: After trabeculectomy with mitomycin C, extremely low intraocular pressure (IOP) with excess filtration may cause hypotonous maculopathy in the early postoperative period. We evaluated the effect of injecting autologous blood on reversing early postoperative marked hypotony after trabeculectomy with mitomycin C. METHODS: Trabeculectomy with mitomycin C was performed in 258 eyes between 1994 and 1998. Peribleb autologous blood injection was performed in five eyes in which pressure patches were ineffective in reversing excess filtration. Approximately 0.1 to 0.3 ml of whole unclotted blood was slowly injected at least 3 mm from the edge of the flap using a sterile 27-gauge needle. RESULTS: None of these eyes developed hypotonous maculopathy after injection. After a mean 31-month follow-up, all eyes had well-controlled IOP and visual acuity in three eyes was much improved. Postoperative complications included mild IOP elevation in one eye treated with laser suturelysis, and fibrinous pupillary membrane in one eye. CONCLUSION: In the early postoperative period, autologous blood injection is effective in reversing excess filtration.

Alpha-helix 2 in the amino-terminal mad homology 1 domain is responsible for specific DNA binding of Smad3.

Smads, signal transducers of the transforming growth factor-beta (TGF-beta) superfamily proteins, directly bind to DNA and regulate transcription of target genes. Smad3 binds to CAGA box, whereas Smad1 and Smad5 preferentially bind to GC-rich sequences. The beta-hairpin loop in the amino-terminal Mad homology 1 (MH1) domain is the direct DNA-binding site of Smad3; however, the amino acid sequences of the beta-hairpin loop of Smad3 and Smad1/5 are identical, suggesting that other regions may be responsible for the differential DNA binding of Smad3 and Smad1/5. To identify regions other than the beta-hairpin loop responsible for specific DNA binding of Smad3, we generated chimeras containing various regions of Smad3 and Smad1. Luciferase assays using a TGF-beta-responsive reporter (CAGA)9-MLP-Luc and gel-mobility shift assays using 3xCAGA as a probe revealed that alpha-helix 2 (H2) in the amino-terminal part of the MH1 domain plays an important role in specific DNA binding and transcriptional activation of Smad3. Luciferase assays using natural TGF-beta-responsive reporters also revealed the functional importance of H2 in the Smad3 MH1 domain in direct DNA binding. Smad3 thus binds to DNA directly through the beta-hairpin loop, and H2 supports specific DNA binding of Smad3.

Mathematical and optimal clustering of test points of the central 30-degree visual field of glaucoma.

PURPOSE: To determine a mathematically optimal sector pattern of the central 30 degree visual field for the follow-up of glaucomatous visual field change based on a large number of actual visual field test data of patients with glaucoma. METHODS: Visual field test data obtained from 1,039 eyes of 1,039 patients with open-angle glaucoma (OAG) using the 30-2 program of the Humphrey Field Analyzer were used for sectorization of the central 30 degree visual field. Of the 1,039 visual field data, 698 (modeling data) were used for determining the sector pattern and 341 (testing data) for checking the sector pattern. The modeling data were further divided into three groups according to the mean deviation (MD) (MD > or = -10 dB, -20 < or = MD < -10 dB, and MD < -20 dB), and the sector pattern was constructed from visual field data of each group using a clustering procedure called VARCLUS. The testing data were used for determining the optimal sector pattern. In a separate set of repeated visual field data of 303 patients with OAG, the fluctuation of MD, sector values of each sector determined, and total deviation of each test point were calculated and compared. RESULTS: The sector pattern constructed from visual field data of MD > or = -10 dB summarized the visual field performance most effectively. The fluctuation of the sector value of each sector was roughly 1.5 times smaller than the total deviation of each test point. CONCLUSION: The sector pattern determined may be useful in analyses of the visual field data of patients with glaucoma.

Role of TGF-beta in the retinoic acid-induced inhibition of proliferation and melanin synthesis in chick retinal pigment epithelial cells in vitro.

The purpose of this study was to investigate the effects of all-trans retinoic acid (RA) on the induction of transforming growth factor-beta (TGF-beta) that is concerned with the proliferation and melanin synthesis of chick retinal pigment epithelial (RPE) cells in vitro. Chick RPE cells were cultured in the presence or absence of RA and anti-TGF-beta antibody for 7 days. The effects of RA and pan-specific TGF-beta antibody on RPE cell proliferation were assessed by counting the number of cells, and their effects on melanin synthesis were evaluated by measuring the melanin content of the cells. TGF-beta activity in the culture supernatant of RPE cells was measured using CCL-64 cells. RA significantly inhibited RPE cell proliferation and increased melanin synthesis. The addition of pan-specific TGF-beta antibody to the culture blocked the inhibition of RPE cell proliferation and the increased melanin synthesis. RA induced TGF-beta production in the culture supernatant of RPE cells. These findings indicate that RA regulates the proliferation and melanin synthesis of RPE cells via induction of TGF-beta.

Results of filtering surgery in young patients with aniridia.

To evaluate whether filtering surgery is effective in controlling the intraocular pressure of young aniridic patients with glaucoma, we retrospectively reviewed the charts of aniridic patients with glaucoma under the age of 40 years. We defined a good intraocular pressure control period as the time from surgery until IOP exceeded 20 mm Hg, with or without glaucoma medication. Twenty filtering surgeries (17 trabeculectomies and 3 trabeculectomies with mitomycin C) were performed on 10 eyes in 6 patients for more than 20 years. The mean good intraocular pressure control period after the filtering surgery was 14.6 months (range, 2 to 54 months). Aside from mild choroidal detachment, no other serious complications were encountered. We believe that filtering surgery is efficacious for control of intraocular pressure of young aniridic patients with glaucoma.

Localization of prostaglandin E receptor subtypes in the ciliary body of mouse eye.

Prostaglandin E2 (PGE2) markedly reduces intraocular pressure (IOP) when applied topically and induces strong relaxation of pre-contracted isolated ciliary muscle through PGE2 receptor. Because the ciliary muscle relaxation reduces IOP by enhancing uveoscleral aqueous outflow, the ciliary muscle where the existence of PGE2 receptors has been demonstrated is thought to be one of the target tissues for PGE2-induced IOP reduction. To investigate the subtypes of PGE2 receptors in the ciliary muscle, the regional distribution of four PGE2 receptor subtypes (EP1, EP2, EP3 and EP4) in the mouse ciliary body was investigated by in situ hybridization using specific probes. Consistent messenger RNA signals for EP1 and EP4 receptors were expressed in the ciliary muscle, although signal levels for these subtypes were less potent as compared with the kidney, which was used as a reference organ. EP2 and EP3 signals were not detected. Stimulation of the EP4 receptor activates adenylate cyclase, which should induce ciliary muscle relaxation. Therefore, the IOP reduction induced by PGE2 analogs may be mediated by the EP4 receptor. In contrast, stimulation of the EP1 receptor is believed to promote intracellular Ca2+ mobilization, and hence should cause ciliary muscle contraction. Thus, the coexistence of EP1 and EP4 receptors in the ciliary muscle suggests that the regulation of ciliary muscle tone by PGE2 is based on a complex mechanism involving multiple receptor subtypes.

Involvement of the protein kinase pathway in melanin synthesis by chick retinal pigment epithelial cells.

Protein kinases are involved in a variety of cellular functions and cell proliferation in eyes. We have explored the involvement of protein kinase C (PKC) in cell proliferation and melanin synthesis by chick retinal pigment epithelial (RPE) cells in vitro. This was achieved by incubation of confluent RPE cells with known inhibitors of protein kinase, H-7, W-7, H-8, and staurosporine. Chick RPE cells were cultured in the presence or absence of the protein kinase inhibitors for a 10-day period. Effects of the inhibitors on cell proliferation and melanin synthesis, as an indication of cell differentiation, were assessed by counting the number of surviving cells and by measuring the melanin content in the cells, respectively. H-7, W-7, and staurosporine inhibited cell proliferation and increased melanin synthesis in a concentration-dependent manner during culture; however, H-8 did not produce these cellular effects. These findings indicate that PKC and calcium/calmodulin-dependent kinase pathways are involved in the proliferation and differentiation of chick RPE cells.

Growth regulation of bovine retinal pericytes by transforming growth factor-beta2 and plasmin.

PURPOSE: Transforming growth factor -beta2 (TGF-beta2) is a predominant isoform of TGF-betas in the eye and plasmin is a peptidase with many functions. To better understand the pathogenesis of retinal microcirculation disorders, the effects of TGF-beta2 and plasmin on cultured bovine retinal pericytes were investigated. METHODS: Exogenous TGF-beta2 or plasmin was added to some cultures, DNA synthesis during cell cycle progression was investigated using [(3)H]thymidine incorporation. Anti-TGF-beta2 antibody was added to neutralize the effects of TGF-beta2. TGF-beta2 in the culture medium was measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Exogenous TGF-beta2 (10 pg to 100 ng/mL) suppressed DNA synthesis. Pericytes produced TGF-beta2. Anti-TGF-beta2 antibody neutralized TGF-beta2 and accelerated DNA synthesis, which shows that pericytes regulate their own cell cycle by action of the autocrine and/or paracrine system of TGF-beta2. Plasmin (0.2 to 0.5 U/mL) accelerated DNA synthesis in a dose-dependent manner, while addition of aprotinin, a protease inhibitor, counteracted this effect of plasmin. The concentration of TGF-beta2 in the culture medium decreased with the addition of plasmin. Simultaneous addition of both plasmin and anti-TGF-beta2 antibody accelerated DNA synthesis. High and low glucose concentrations of the culture medium did not affect DNA synthesis. CONCLUSIONS: Our results suggest that TGF-beta2 and plasmin respectively decrease and increase DNA synthesis. In a retinal microcirculation disorder, they may play competitive roles in the cell cycle of pericytes.

Efficacy and safety of latanoprost eye drops for glaucoma treatment: a 1-year study in Japan.

PURPOSE: To evaluate the intraocular pressure (IOP)-lowering effect and safety of latanoprost, a prostaglandin analogue, in patients with primary open-angle glaucoma or ocular hypertension. METHOD: One hundred and twenty-four Japanese patients with primary open-angle glaucoma or ocular hypertension were enrolled in this open-labeled study and were treated with 0.005% latanoprost once daily for 1 year. RESULTS: At all follow-up visits there was a significant (P < .001) reduction in IOP compared with the baseline value. After 1 year, the IOP was reduced by 5.4 +/- 2.9 (mean +/- SD) mm Hg from a baseline value of 23.5 +/- 2.2 mm Hg. No evidence of an upward drift in the IOP was observed during the treatment period. The most frequently reported adverse ocular events were mild conjunctival hyperemia and iris pigmentation. Very few adverse systemic events were observed. CONCLUSIONS: Latanoprost eye drops showed a marked and stable IOP-lowering effect during the 1-year treatment period. Furthermore, latanoprost was well-tolerated and should be a valuable contribution to the management of glaucoma.

Characterization of a bone morphogenetic protein-responsive Smad-binding element.

Bone morphogenetic proteins (BMPs) are pleiotropic growth and differentiation factors belonging to the transforming growth factor-beta (TGF-beta) superfamily. Signals of the TGF-beta-like ligands are propagated to the nucleus through specific interaction of transmembrane serine/threonine kinase receptors and Smad proteins. GCCGnCGC has been suggested as a consensus binding sequence for Drosophila Mad regulated by a BMP-like ligand, Decapentaplegic. Smad1 is one of the mammalian Smads activated by BMPs. Here we show that Smad1 binds to this motif upon BMP stimulation in the presence of the common Smad, Smad4. The binding affinity is likely to be relatively low, because Smad1 binds to three copies of the motif weakly, but more repeats of the motif significantly enhance the binding. Heterologous reporter genes (GCCG-Lux) with multiple repeats of the motif respond to BMP stimulation but not to TGF-beta or activin. Mutational analyses reveal several bases critical for the responsiveness. A natural BMP-responsive reporter, pTlx-Lux, is activated by BMP receptors in P19 cells but not in mink lung cells. In contrast, GCCG-Lux responds to BMP stimulation in both cells, suggesting that it is a universal reporter that directly detects Smad phosphorylation by BMP receptors.