Effect of topical prostaglandins on the biomechanics and shape of the cornea.

PURPOSE: To investigate the effect of topical prostaglandin (PG) eyedrops on the biomechanics and shape of the cornea. METHODS: Consecutive patients with primary open-angle glaucoma who had been treated with the same anti-glaucoma PG (114 eyes, 57 patients) or ß-blocker (36 eyes, 18 patients) eyedrops in 1 eye alone for more than 3 months at Inouye Eye Hospital or Miyata Eye Hospital. The biomechanical properties of the cornea were measured using a Corvis ST device (Oculus, Wetzlar, Germany). Corneal tomography was measured with a Casia 1 or 2 (Tomey Corp., Nagoya, Japan). The biomechanical parameters and tomographic features of the cornea were compared between PG-treated eyes, ß-blocker-treated eyes, and contralateral eyes using a mixed-effect model adjusted for intraocular pressure and corneal thickness and a linear model adjusted for age, sex, intraocular pressure, and corneal thickness. RESULTS: The PG-treated eyes had a significantly smaller applanation 1 time, the highest concavity radius, and a larger deformation amplitude than the contralateral eyes. Also, the PG-treated eyes had a significantly smaller applanation 1 time and applanation 2 velocity but a larger peak distance than the ß-blocker-treated eyes. There were no significant differences in any of the Casia parameters between the PG-treated eyes and the contralateral eyes. CONCLUSION: The results of this study suggest that topical PG makes biochemical changes to the cornea but does not alter its shape.

2-O-Sulfated Domains in Syndecan-1 Heparan Sulfate Inhibit Neutrophil Cathelicidin and Promote Staphylococcus aureus Corneal Infection.

Ablation of syndecan-1 in mice is a gain of function mutation that enables mice to significantly resist infection by several bacterial pathogens. Syndecan-1 shedding is induced by bacterial virulence factors, and inhibition of shedding attenuates bacterial virulence, whereas administration of purified syndecan-1 ectodomain enhances virulence, suggesting that bacteria subvert syndecan-1 ectodomains released by shedding for their pathogenesis. However, the pro-pathogenic functions of syndecan-1 ectodomain have yet to be clearly defined. Here, we examined how syndecan-1 ectodomain enhances Staphylococcus aureus virulence in injured mouse corneas. We found that syndecan-1 ectodomain promotes S. aureus corneal infection in an HS-dependent manner. Surprisingly, we found that this pro-pathogenic activity is dependent on 2-O-sulfated domains in HS, indicating that the effects of syndecan-1 ectodomain are structure-based. Our results also showed that purified syndecan-1 ectodomain and heparan compounds containing 2-O-sulfate motifs inhibit S. aureus killing by antimicrobial factors secreted by degranulated neutrophils, but does not affect intracellular phagocytic killing by neutrophils. Immunodepletion of antimicrobial factors with staphylocidal activities demonstrated that CRAMP, a cationic antimicrobial peptide, is primarily responsible for S. aureus killing among other factors secreted by degranulated neutrophils. Furthermore, we found that purified syndecan-1 ectodomain and heparan compounds containing 2-O-sulfate units potently and specifically inhibit S. aureus killing by synthetic CRAMP. These results provide compelling evidence that a specific subclass of sulfate groups, and not the overall charge of HS, permits syndecan-1 ectodomains to promote S. aureus corneal infection by inhibiting a key arm of neutrophil host defense.

Long-term follow-up of transplantation of preserved limbal allograft and amniotic membrane for recurrent pterygium.

PURPOSE: To conduct a long-term follow-up study evaluating the efficacy and safety of transplantation of preserved limbal allograft and amniotic membrane for recurrent pterygium. METHODS: This was a retrospective, non-comparative, interventional case series conducted at a private eye hospital. Eighty-four eyes of 80 patients with recurrent pterygium were included in the study. The mean number of previous surgeries for pterygium was 1.36 ± 0.98 (range, 1-8). All subjects received transplantation of preserved limbal allograft and amniotic membrane. RESULTS: The mean follow-up period was 73.0 ± 38.1 months (range, 12-154 months). Pterygium recurred in 10 eyes (11.9 %). The mean period to recurrence was 16.3 ± 11.3 months (range, 5-33 months). Symblepharon was cured in 21 eyes, persisted in 2 eyes, and newly occurred in 3 eyes. Diplopia was cured in eight eyes, persisted in five eyes, and newly occurred in one eye. As for complications, intraocular pressure elevations over 21 mmHg were recognized in ten eyes of nine cases, in which the intraocular pressure was controlled by reduction of topical steroid in four eyes and by addition of topical prostaglandin derivatives in six eyes. Twenty-four eyes (28.6 %) gained two lines or more of Landolt best spectacle-corrected visual acuity (BSCVA), 56 eyes (66.7 %) stayed within one line from preoperation, and four eyes (4.8 %) lost two lines or more. There were no major complications and no graft rejection. CONCLUSIONS: Transplantation of preserved limbal allograft and amniotic membrane is a safe and effective procedure for recurrent pterygium.

Systemic vascular safety of ranibizumab for age-related macular degeneration: systematic review and meta-analysis of randomized trials.

BACKGROUND: We conducted a meta-analysis of randomized trials of ranibizumab for age-related macular degeneration (AMD) to elucidate systemic vascular risk. CLINICAL RELEVANCE: Although intravitreal vascular endothelial growth factor inhibitors are widely used to treat AMD, whether they produce systemic adverse effects remains uncertain. METHODS: We searched MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials through March 2014 to identify the randomized trials that compared systemic safety among different intensities of ranibizumab treatment for AMD. The outcome measures were the incidence of cerebrovascular accidents (CVAs), myocardial infarctions, nonocular hemorrhages, overall arterial thromboembolic events (ATEs), and all-cause mortality. We calculated the Peto odds ratio (OR) with 95% confidence interval for the comparisons between different intensities of regimens in terms of dose and retreatment frequency. RESULTS: Eleven trials comprising 6596 patients with AMD were included in the meta-analysis. A significant increase was observed in the following comparisons: 0.5 versus 0.3/0.0 mg for CVA (OR, 1.86; 95% CI, 1.05-3.29; P = 0.03), monthly versus pro re nata (PRN)/0.0 mg for CVA (OR, 1.89; 95% CI, 1.06-3.38; P = 0.03), and 0.3/0.5 versus 0.0 mg for nonocular hemorrhage (OR, 1.57; 95% CI, 1.01-2.44; P = 0.04). A nonsignificant increase was observed in the following comparisons: 0.5 versus 0.0 mg for CVA (OR, 2.27; 95% CI, 0.90-5.69; P = 0.08), monthly versus PRN for CVA (OR, 2.04; 95% CI, 0.94-4.45; P = 0.07), 0.5 versus 0.0 mg for nonocular hemorrhage (OR, 1.68; 95% CI, 0.98-2.88; P = 0.06), 0.3 versus 0.0 mg for nonocular hemorrhage (OR, 1.68; 95% CI, 0.95-2.98; P = 0.07), monthly versus PRN/0.0 mg for nonocular hemorrhage (OR, 1.54; 95% CI, 0.98-2.42; P = 0.06), monthly versus PRN for ATE (OR, 1.58; 95% CI, 0.96-2.61; P = 0.07), and monthly versus PRN/0.0 mg for ATE (OR, 1.42; 95% CI, 0.99-2.05; P = 0.06). Among the other analyses, no protective or harmful effects of ranibizumab were observed. CONCLUSIONS: In ranibizumab treatment for patients with AMD, a possible relationship of more intensive treatment to more systemic vascular adverse events was identified, but no relationship with mortality was identified.

[Grading for corneal endothelial damage].

While corneal endothelial damage in its most advanced stage results in bullous keratopathy where corneal transplantation is needed, there is no severity grading for corneal endothelial damage. The Japanese Corneal Society formed a working group to establish a severity grading for corneal endothelial damage. The following are the proposed severity grading criteria. Normal: Corneal endothelial cell (CEC) density is 2000 cells/mm2 or more. Grade 1: CEC density between 1000 and 2000 cells/ mm2. The corneal endothelium is below the normal physiological condition. Grade 2: CEC density between 500 and 1000 cells/ mm2. The transparency of the corneal endothelium is endangered. Even slight damage can induce bullous keratopathy. Grade 3: CEC density below 500 cells/mm2 but no corneal edema is observed. Grade 4: Bullous keratopathy. The cornea is opaque with edema.

Presynaptic dystroglycan-pikachurin complex regulates the proper synaptic connection between retinal photoreceptor and bipolar cells.

Dystroglycan (DG) is a key component of the dystrophin-glycoprotein complex (DGC) at the neuromuscular junction postsynapse. In the mouse retina, the DGC is localized at the presynapse of photoreceptor cells, however, the function of presynaptic DGC is poorly understood. Here, we developed and analyzed retinal photoreceptor-specific DG conditional knock-out (DG CKO) mice. We found that the DG CKO retina showed a reduced amplitude and a prolonged implicit time of the ERG b-wave. Electron microscopic analysis revealed that bipolar dendrite invagination into the photoreceptor terminus is perturbed in the DG CKO retina. In the DG CKO retina, pikachurin, a DG ligand in the retina, is markedly decreased at photoreceptor synapses. Interestingly, in the Pikachurin(-/-) retina, the DG signal at the ribbon synaptic terminus was severely reduced, suggesting that pikachurin is required for the presynaptic accumulation of DG at the photoreceptor synaptic terminus, and conversely DG is required for pikachurin accumulation. Furthermore, we found that overexpression of pikachurin induces formation and clustering of a DG-pikachurin complex on the cell surface. The Laminin G repeats of pikachurin, which are critical for its oligomerization and interaction with DG, were essential for the clustering of the DG-pikachurin complex as well. These results suggest that oligomerization of pikachurin and its interaction with DG causes DG assembly on the synapse surface of the photoreceptor synaptic terminals. Our results reveal that the presynaptic interaction of pikachurin with DG at photoreceptor terminals is essential for both the formation of proper photoreceptor ribbon synaptic structures and normal retinal electrophysiology.

Prevalence of dry eye syndrome and risk factors in physical education and sports science graduates.

PURPOSE: There are only a few epidemiological studies of dry eye syndrome (DES) in populations with a common academic background. in this study, the prevalence of DES and associated factors were evaluated separately in men and women physical education and sports science graduates. STUDY DESIGN: Cross-sectional survey. METHODS: A questionnaire about the diagnosis of DES and associated factors was mailed to 9507 graduates of the Faculty of Health and Sports Science, Juntendo University. The questions covered subjective DES using Schaumberg et al.'s questionnaire and the prevalence of diagnosed DES. Associated factors, age, sex, smoking, alcohol consumption, body mass index, daily screen viewing time, and contact lens (CL) use were analyzed. RESULTS: A total of 2048 valid responses were received. The prevalence of diagnosed DES was 2.9% in men and 9.3% in women. For subjective DES, the prevalence was 14.8% in men and 39.8% in women. The odds ratio for DES was high in men and women who used CLs and women whose daily screen viewing time was ≥ 4 h. CONCLUSION: Both diagnosed and subjective DES were highly prevalent in men and women of all ages, particularly among those in their 20 and 30s. CL use was associated with DES in both men and women. Measures to deal with the factors that can be corrected might have a positive effect on the ocular health and quality of life of physical education and sports science graduates.

Syndecan-1 promotes Staphylococcus aureus corneal infection by counteracting neutrophil-mediated host defense.

Many microbial pathogens subvert cell surface heparan sulfate proteoglycans (HSPGs) to infect host cells in vitro. The significance of HSPG-pathogen interactions in vivo, however, remains to be determined. In this study, we examined the role of syndecan-1, a major cell surface HSPG of epithelial cells, in Staphylococcus aureus corneal infection. We found that syndecan-1 null (Sdc1(-/-)) mice significantly resist S. aureus corneal infection compared with wild type (WT) mice that express abundant syndecan-1 in their corneal epithelium. However, syndecan-1 did not bind to S. aureus, and syndecan-1 was not required for the colonization of cultured corneal epithelial cells by S. aureus, suggesting that syndecan-1 does not mediate S. aureus attachment to corneal tissues in vivo. Instead, S. aureus induced the shedding of syndecan-1 ectodomains from the surface of corneal epithelial cells. Topical administration of purified syndecan-1 ectodomains or heparan sulfate (HS) significantly increased, whereas inhibition of syndecan-1 shedding significantly decreased the bacterial burden in corneal tissues. Furthermore, depletion of neutrophils in the resistant Sdc1(-/-) mice increased the corneal bacterial burden to that of the susceptible WT mice, suggesting that syndecan-1 moderates neutrophils to promote infection. We found that syndecan-1 does not affect the infiltration of neutrophils into the infected cornea but that purified syndecan-1 ectodomain and HS significantly inhibit neutrophil-mediated killing of S. aureus. These data suggest a previously unknown bacterial subversion mechanism where S. aureus exploits the capacity of syndecan-1 ectodomains to inhibit neutrophil-mediated bacterial killing mechanisms in an HS-dependent manner to promote its pathogenesis in the cornea.

Clinical features and haplotype analysis of newly identified Japanese patients with gelsolin-related familial amyloidosis of Finnish type.

Familial amyloidosis of the Finnish type (FAF) is an autosomal dominant form of systematic amyloidosis characterized by lattice corneal dystrophy, cranial neuropathy, and cutis laxa. Although FAF has been frequently found in the Finnish population, FAF is a considerably rare disorder in other regions. In this study, we examined the clinical characteristics as well as the haplotypes of six Japanese patients with FAF from five families. They showed the typical clinical presentations of FAF, but we found a broad range of ages at onset of neurological symptoms. All members had the c.654G>A mutation in GSN. To evaluate the disease haplotypes, high-density single-nucleotide polymorphism (SNP) arrays were used and disease-relevant haplotypes were reconstructed. Haplotype analysis in the four apparently unrelated families suggested a common founder haplotype. In a sporadic FAF patient, however, the haplotype was dissimilar to the founder haplotype. The present study demonstrated that a founder mutation in most of the Japanese families with FAF, except for a sporadic patient in whom a de novo mutation event was suggested as the origin of the mutation.

Caffeine increases tear volume depending on polymorphisms within the adenosine A2a receptor gene and cytochrome P450 1A2.

PURPOSE: The primary aim of the present study was to examine the effect of caffeine on tear volume. The secondary aim was to investigate the relation between caffeine-induced changes in tear volume and polymorphisms in ADORA2A and CYP1A2. DESIGN: Double-masked, placebo-controlled, crossover study. PARTICIPANTS: Seventy-eight healthy volunteers were recruited for the study. METHODS: Subjects participated in 2 sessions in which they received capsules containing either placebo or caffeine. The caffeine capsules were given to the subjects to keep the caffeine volume per body weight within 5 to 7 mg/kg. After caffeine intake, tear meniscus height (TMH) was measured. Subjects provided a blood sample for genotyping. MAIN OUTCOME MEASURES: Tear meniscus height, single nucleotide polymorphism. RESULTS: The tear volume increased after caffeine consumption. The net increase in TMH was 0.08 mm (95% confidence interval, 0.05-0.10) greater when participants were given caffeine than when given placebo (P<0.0001). In ADORA2A, the difference in the net increase in TMH for participants who were heterozygous at rs5751876 and rs2298383 was 0.07 mm (P = 0.001) and who were minor homozygous was 0.08 mm (P = 0.007). In CYP1A2, the net increase in TMH for participants who were minor homozygous at rs2472304 was lower than for those who were major homozygous; the difference was 0.06 mm (P = 0.039). CONCLUSIONS: Caffeine intake increases tear volume and polymorphisms within ADORA2A, and CYP1A2 is associated with the tear increase after caffeine intake. Genetic polymorphisms had a significant effect on tear meniscus that was of limited clinical significance.

Effects of long-term topical anti-glaucoma medications on meibomian glands.

OBJECTIVE: To examine effects of long-term topical anti-glaucoma medications on meibomian gland morphology and function and assess their relationship with slit-lamp findings. METHODS: This was a cross-sectional observational case series of 31 patients with glaucoma (mean age ± standard deviation, 65.0 ± 13.0 years; mean duration of eye drop use, 7.9 ± 6.0 years) treated with topical anti-glaucoma drugs in only one eye for more than 1 year: 13 receiving prostaglandin analogues (PGs) alone, eight receiving ß-blockers alone, and ten receiving multiple treatments. Untreated contralateral eyes served as controls. Lid margin (lid margin abnormality score: 0-4) and superficial punctate keratopathy (SPK score: 0-1) were observed with a slit lamp. Upper and lower eyelids were turned over to observe meibomian glands using non-contact meibography. Meibomian gland loss was scored for each eyelid from grade 0 (no loss of meibomian glands) through grade 3 (loss >2/3 of total meibomian gland area). Meibomian lipid content (meibum) was scored (meibum score: 0-3). RESULTS: Treated eyes had significantly higher scores for lid margin abnormality (P= 0.001), SPK (P< 0.001), meibo-score (P< 0.001), and meibum (P< 0.001) than control eyes. Tear film break-up time (BUT) was significantly shorter in treated eyes than in control eyes (P= 0.001). Schirmer values were significantly lower in treated eyes than in control eyes (P= 0.0039). Subgroup analysis indicated a significantly higher meibo-score in eyes treated with PGs (P= 0.0046) and in eyes treated with ß-blockers (P= 0.0231) than in the corresponding controls. CONCLUSIONS: Long-term anti-glaucoma eye drop use affects meibomian gland morphology and function.

Specific tear IgE in patients with moderate-to-severe autumnal allergic conjunctivitis.

BACKGROUND: Recently, a new commercial test for specific IgE based on immunochromatography (Immfast Check J1®) has been developed. We previously reported on the use of this kit for tear fluid samples during spring. In this study, we compared the level of specific IgE in tear fluid among patients with seasonal allergic conjunctivitis during spring or autumn and control subjects. METHODS: A nonrandomized, cross-sectional study was conducted in patients with moderate-to-severe allergic conjunctivitis occurring in spring (n = 56, spring group) or autumn (n = 52, autumn group), as well as in age- and sex-matched healthy control subjects (n = 50, control group). Levels of specific IgE for cedar pollen, cat epithelium and Dermatophagoides pteronyssinus were measured in tear fluid with the Immfast Check J1. RESULTS: Specific IgE scores for cedar pollen and D. pteronyssinus were significantly higher in the spring and autumn groups than in the control group (p < 0.0001). In the autumn group, the highest detection rate for a specific allergen was 96.2% for D. pteronyssinus, followed by cedar pollen (86.5%) and cat epithelium (25.5%). In the spring group, the highest detection rate was 98.3% for cedar pollen, followed by D. pteronyssinus (51.7%) and cat epithelium (19.0%). CONCLUSIONS: These results suggest that both house dust mite allergen and outdoor allergens such as cedar pollen can be causes of allergic conjunctivitis during both spring and autumn in Japan. The Immfast Check J1 provides rapid measurement of specific IgE in tear fluid, which allows easy diagnosis of allergic conjunctivitis in an outpatient setting.

Relationship between total tear and serum IgE in allergic conjunctivitis.

BACKGROUND: Recently, a new commercial test for total tear IgE based on immunochromatography (Allerwatch®) was developed. We examined the relationship between total tear and serum IgE levels with the Allerwatch test. METHODS: A nonrandomized cross-sectional study was conducted in 40 patients with allergic conjunctivitis, 35 age- and sex-matched healthy control subjects, and 6 patients with epidemic keratoconjunctivitis (EKC). The total tear IgE score was determined with the Allerwatch test (0, 1 and 2), and the serum total IgE level was measured by solid-phase immunoassay (Phadezym PRIST) in all subjects. RESULTS: Total tear IgE was assayed within 10 min of collection in all samples. The total tear IgE-positive rate was significantly higher in the allergic group than in the control and EKC groups (92.5 vs. 0.0 vs. 0.0%; p < 0.00001). Sensitivity and specificity were 0.925 and 1.000, respectively. The total IgE score and log (total serum IgE) were also higher in the allergic group than in the control and EKC groups [total IgE score: 1.48 ± 0.63 vs. 0.00 ± 0.00 vs. 0.00 ± 0.00, p < 0.00001; log (serum total IgE): 2.23 ± 0.50 vs. 0.80 ± 0.31 vs. 0.79 ± 0.37, p < 0.00001]. The total tear IgE score was significantly correlated with log (total serum IgE) in the allergic group (r = 0.712, p < 0.00001). CONCLUSIONS: These results show that this rapid test for the measurement of total tear IgE is easy to perform on an outpatient basis and may be helpful in the management of ocular allergy.

Limbal relaxing incisions using a reference point and corneal topography for intraoperative identification of the steepest meridian.

PURPOSE: To examine the efficacy and safety of topography-based limbal relaxing incision (LRI) surgery. METHODS: Forty-four eyes of 36 consecutive patients who underwent cataract surgery more than 1 month previously, had refractive against-the-rule astigmatism of ≥2.00 diopters (D), and were scheduled to undergo LRI surgery were enrolled in the study. Patients were randomized into two groups-a topography-based LRI group (19 eyes of 14 patients) and a conventional LRI group (25 eyes of 22 patients). The topography-based LRI procedure comprised 3 steps: placing a mark on the cornea and conjunctiva, identifying this mark in the topographic image, and performing LRI based on the location of the mark. In the conventional LRI group, the horizontal meridian was marked under a slit lamp and LRIs were made based on the position of the horizontal mark. RESULTS: Corneal astigmatism in the topography-based LRI group before and 1 month after surgery was 2.03±0.92 D and 1.33±0.69 D, respectively (P=.014). Corneal astigmatism in the conventional LRI group before and 1 month after surgery was 2.36±0.77 D and 0.93±0.70 D, respectively (P<.0001). Fourier harmonic analysis of the topography data demonstrated that regular astigmatism was significantly decreased in the two groups. The mean regular astigmatism was not significantly different before and after LRI between groups, whereas the variances (mean of the deviation squared from its mean) of regular astigmatism were significantly different between groups 1 month after LRI. CONCLUSIONS: Compared with conventional LRI surgery, a topography-based procedure may reduce the deviation of the effect of LRIs.

Intraoperative mydriasis by intracameral injection of mydriatic eye drops: in vivo efficacy and in vitro safety studies.

BACKGROUND: This study investigated the efficacy and safety of intracameral injection of commercially available eye drops containing 0.5% tropicamide and 0.5% phenylephrine hydrochloride (Mydrin-P, Santen Pharmaceutical, Osaka, Japan). DESIGN: In vitro experiment and prospective clinical study at a private hospital. PARTICIPANTS AND SAMPLES: Mydrin-P was applied to confluent cultured human corneal endothelial cells, and the cellular morphology was examined. Clinical study subjects were 65 eyes of 65 patients that underwent phaco-emulsification and aspiration with intraocular lens implantation and received intracameral injection of Mydrin-P for poor mydriasis after preoperative topical instillation of mydriatics (intraocular mydriasis group; with five subgroups based on cause: diabetes, pseudo-exfoliation, post-surgery, uveitis, unknown). Controls, comprising 39 eyes of 39 patients, were not injected with Mydrin-P. METHODS: The ratio of pupillary diameter to corneal diameter was determined before and after injection of Mydrin-P. Corneal endothelial density was measured preoperatively and 3 months and 1 year postoperatively. MAIN OUTCOME MEASURES: Pupillary diameter and corneal endothelial density. RESULTS: Human corneal endothelial cell morphology was unaltered after Mydrin-P injection. The mean ratio of the pupillary diameter to corneal diameter increased in the intraocular mydriasis group (before: 54.2 ± 4.8%, after: 58.4 ± 6.6%; P < 0.001) and in the diabetes and unknown subgroups. The corneal endothelial cell density reduction rate 3 months and 1 year after surgery was not significantly different between the intraocular mydriasis group and controls. CONCLUSION: Intracameral injection of Mydrin-P appears to be effective and safe for dilating the pupil in cases with poor mydriasis after preoperative instillation of mydriatics.

Severity and determinants of pinguecula in a hospital-based population.

OBJECTIVES: Pinguecula is a relatively frequent disease; however, there have been no reports about the grade of pinguecula. We investigated the relationship between age and the prevalence and severity of pinguecula by using a grading system and compared the grade of pinguecula between men and women and between the nasal and temporal halves of the conjunctiva. METHODS: This was a prospective, randomized study of 1,040 patients aged 1 to 95 years, including 520 men and 520 age-matched women. The age, gender, medical history, ocular history, and grade of pingueculae located on the nasal and temporal conjunctiva were determined in all subjects. RESULTS: The prevalence of pinguecula increased dramatically with age. The mean grade was higher in male patients than in female patients for pingueculae located on both the nasal conjunctiva (P = 0.00072) and the temporal conjunctiva (P = 0.00035). However, there was no significant difference in the grade of pinguecula between the nasal and temporal conjunctiva. CONCLUSIONS: This was the first assessment of the grade of pinguecula in a large series of patients. Our findings strongly suggest that pinguecula is an age-dependent lesion that occurs from young adulthood onward. The present grading system should also be useful for future clinical studies of pinguecula.

Relationship between conjunctivochalasis and refractive error.

OBJECTIVES: To assess the relation between the prevalence and grade of conjunctivochalasis and refractive error and to compare the grade of conjunctivochalasis between myopic and hyperopic patients. METHODS: Consecutive patients aged from 3 to 94 years were chosen for this study. Exclusion criteria included a history of using contact lenses, ocular surgeries, infectious conjunctivitis, or corneal diseases. The age, gender, medical history, ocular history, the grade and other parameters of inferior conjunctivochalasis classified into three locations (nasal, middle, and temporal), and refractive error were determined in all subjects. Patients were divided into three groups as follows: a hyperopic group (≥0.0 D), an emmetropic group (<0.0 and ≥-2.0 D), and a myopic group (<-2.0 D). They were also divided into 10 groups according to age. One-way analysis of variance and the Scheffe multiple comparison test were used to compare the mean values among three groups. Relations among the variables were investigated by calculating Pearson correlation coefficients and partial correlation coefficients. RESULTS: A total of 1,110 patients were included in the study. In each age group, the mean grade of conjunctivochalasis was higher in hyperopic patients than in myopic patients. There were no significant differences in both the downward gaze- and digital pressure-dependent changes of conjunctivochalasis between the myopic and hyperopic groups. The severity of conjunctivochalasis affecting the nasal and temporal bulbar conjunctiva, and parameters such as the changes of conjunctivochalasis caused by downward gaze or digital pressure, were correlated with the refractive error, especially in patients over 40 years old (P<0.05). CONCLUSIONS: This was the first assessment of the relationship between refractive error and the grade of conjunctivochalasis in a large consecutive series of patients. Our results suggest that the prevalence and grade of conjunctivochalasis are dependent on refractive error, with hyperopia being an important risk factor for conjunctivochalasis.

Laser treatment of giant iris cyst with nanophthalmos.

We report a case of a giant iris cyst treated with neodymium-doped yttrium aluminum garnet laser cystotomy. A 33-year-old woman presented with nanophthalmos and ocular hypertension secondary to a giant iris cyst after congenital cataract surgery in the left eye. Examination with a rotating Scheimpflug camera and anterior-segment optical coherence tomography revealed a large iris cyst surrounding the pupillary region. She underwent laser iridocystotomy and irrigation of the anterior chamber. No recurrence of the iris cyst has been noted after 18 months and the IOP is normal. The cyst was treated successfully by laser cystotomy without other ocular tissue damage. Examination with a rotating Scheimpflug camera and anterior-segment optical coherence tomography proved to be simple and quick methods for assessing the iris cyst.

VEGF164-mediated inflammation is required for pathological, but not physiological, ischemia-induced retinal neovascularization.

Hypoxia-induced VEGF governs both physiological retinal vascular development and pathological retinal neovascularization. In the current paper, the mechanisms of physiological and pathological neovascularization are compared and contrasted. During pathological neovascularization, both the absolute and relative expression levels for VEGF164 increased to a greater degree than during physiological neovascularization. Furthermore, extensive leukocyte adhesion was observed at the leading edge of pathological, but not physiological, neovascularization. When a VEGF164-specific neutralizing aptamer was administered, it potently suppressed the leukocyte adhesion and pathological neovascularization, whereas it had little or no effect on physiological neovascularization. In parallel experiments, genetically altered VEGF164-deficient (VEGF120/188) mice exhibited no difference in physiological neovascularization when compared with wild-type (VEGF+/+) controls. In contrast, administration of a VEGFR-1/Fc fusion protein, which blocks all VEGF isoforms, led to significant suppression of both pathological and physiological neovascularization. In addition, the targeted inactivation of monocyte lineage cells with clodronate-liposomes led to the suppression of pathological neovascularization. Conversely, the blockade of T lymphocyte-mediated immune responses with an anti-CD2 antibody exacerbated pathological neovascularization. These data highlight important molecular and cellular differences between physiological and pathological retinal neovascularization. During pathological neovascularization, VEGF164 selectively induces inflammation and cellular immunity. These processes provide positive and negative angiogenic regulation, respectively. Together, new therapeutic approaches for selectively targeting pathological, but not physiological, retinal neovascularization are outlined.

Isolation of adult progenitor cells with neuronal potential from rabbit corneal epithelial cells in serum- and feeder layer-free culture conditions.

PURPOSE: To isolate progenitor cells from rabbit corneal epithelial cells (CEC) in serum- and feeder layer-free culture conditions and to compare the self-renewal capacity of corneal epithelial progenitor cells obtained from the central and limbal regions of the cornea. METHODS: Tissue samples of New Zealand white rabbit corneas were dissected from the limbal and central regions to obtain CEC for sphere-forming culture, in which the cells formed spheres in serum-free medium containing growth factors. The number of primary and secondary sphere colonies and the size of the primary spheres were compared between the limbal and central regions. To promote differentiation, isolated sphere colonies were plated in dishes coated with poly-L-lysine (PLL)/laminin. The expression of epithelial, neural, and mesenchymal mRNAs was examined in the sphere colonies and their progeny by immunocytochemistry and/or the reverse transcription-polymerase chain reaction (RT-PCR). Adherent differentiated cells from the sphere colonies were also examined morphologically. RESULTS: Primary spheres were isolated from both the limbal and central regions of the cornea. The rate of primary sphere formation by CEC from the limbal region (55.6+/-10.6/10,000 cells) was significantly higher than that by cells from the central cornea (43.1+/-7.2/10,000 cells, p=0.0028), but there was no significant difference in the size of primary spheres derived from both regions. The self-renewal capacity of cells from the limbal region was higher than that of cells from the central region, as evidenced by the significantly higher secondary sphere formation rate of limbal cells (38.7+/-8.5/10,000 cells) in comparison with that for central cells (31.3+/-5.7/10,000 cells, p=0.013). The primary sphere colonies expressed bromodeoxyuridine (BrdU), a 63-kDa protein (p63), p75 neurotrophin receptor (p75(NTR)), and nestin, whereas their progeny expressed cytokeratin 3, cytokeratin 12, vimentin, alpha-smooth muscle actin, microtubule-associated protein 2, and neuron-specific enolase on immunocytochemical analysis. These markers were confirmed by RT-PCR. CONCLUSIONS: Our findings indicate that limbal CEC contain more progenitor cells with a stronger self-renewal capacity than cells from the central region. These progenitor cells differentiate into the epithelial lineage, and can also produce neuronal protein.