Neurotoxicity screening in a multipotent neural stem cell line established from the mouse brain.

Neural stem cells (NSCs) have mainly been applied to neurodegeneration in some medically intractable neurologic diseases. In this study, we established a novel NSC line and investigated the cytotoxic responses of NSCs to exogenous neurotoxicants, glutamates and reactive oxygen species (ROS). A multipotent NSC line, B2A1 cells, was established from long-term primary cultures of oligodendrocyte-enriched cells from an adult BALB/c mouse brain. B2A1 cells could be differentiated into neuronal, astrocytic and oligodendroglial lineages. The cells also expressed genotypic mRNA messages for both neural progenitor cells and differentiated neuronoglial cells. B2A1 cells treated with hydrogen peroxide and L-buthionine-(S,R)-sulfoximine underwent 30-40% cell death, while B2A1 cells treated with glutamate and kainate showed 25-35% cell death. Cytopathologic changes consisting of swollen cell bodies, loss of cytoplasmic processes, and nuclear chromatin disintegration, developed after exposure to both ROS and excitotoxic chemicals. These results suggest that B2A1 cells may be useful in the study of NSC biology and may constitute an effective neurotoxicity screening system for ROS and excitotoxic chemicals.

Brown syndrome with bifid scleral insertion of the superior oblique.

The authors describe a patient with bilateral congenital Brown syndrome who had bifid scleral insertion of the superior oblique muscle of both eyes. They considered bifid scleral insertion of the superior oblique muscle as a possible cause of congenital Brown syndrome.

Intravitreal triamcinolone reduces the morphologic changes of ciliary body after pars plana vitrectomy for retinal vascular diseases.

PURPOSE: To investigate the early postoperative changes in ciliary body after pars plana vitrectomy (PPV) for retinal vascular disease and to determine whether intravitreal triamcinolone acetonide (IVTA) affects postvitrectomy changes of the ciliary body. DESIGN: Prospective interventional case-control study. METHODS: We recruited 46 patients who underwent uncomplicated PPV for retinal vascular disease (35 with proliferative diabetic retinopathy (PDR) and 11 with branch retinal vein occlusion) and divided the patients into two groups according to the use of IVTA at the end of the PPV. The morphologic changes of the anterior segments were measured by ultrasound biomicroscopy (UBM) one day before and one day, two days, three days, five days, two weeks, one month, and two months after the PPV. The main outcome measures were the thickness and area of the ciliary body, the frequency of supraciliary effusions (SEs), angle-opening, and anterior chamber depth (ACD). We compared the UBM parameters between the two groups. RESULTS: The thickness and area of the ciliary body significantly increased from day 1 to day 5 postoperatively. Nineteen of 46 eyes had SEs that were frequently associated with divergent alterations of postoperative intraocular pressure (IOP). The angle-opening and ACD were significantly decreased and dependent upon the findings of ciliary body thickness and SEs. The degree of the morphologic changes of the ciliary body and the frequency of SEs were significantly lower and of shorter duration in the IVTA group compared to the non-IVTA group. CONCLUSIONS: The PPV for retinal vascular disease induces the morphologic changes of the ciliary body associated with postoperative abnormalities in IOP during the early postoperative period. Administration of IVTA at the end of the PPV effectively reduces or shortens the duration of the postvitrectomy changes of the ciliary body.

The effect of surgical correction of epiblepharon on astigmatism in children.

PURPOSE: To evaluate the characteristics of astigmatism in epiblepharon and postoperative astigmatic changes in children. METHODS: The authors undertook a retrospective chart review of 56 patients who underwent an epiblepharon operation. Preoperative astigmatism was compared with postoperative astigmatism according to age at surgery, the presence of amblyopia, and unilateral epiblepharon. RESULTS: Astigmatism decreased from a preoperative mean of 1.34 +/- 1.25 D with-the-rule astigmatism to a postoperative mean of 1.10 +/- 1.14 D with-the-rule astigmatism in all patients (P = .055), and this reduction was statistically significant in the 5- to 7-year age group (P = .038), amblyopic eyes (P = .021), and affected eyes in cases of unilateral epiblepharon (P = .004). CONCLUSIONS: Epiblepharon operations were found to change astigmatism. The results obtained suggest that an epiblepharon operation should be considered in patients with amblyopia and epiblepharon prior to the general treatment of amblyopia.

Application of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis.

PURPOSE: To investigate the efficacy of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Twenty-eight eyes of 28 patients with neurotrophic keratitis who were refractory to conventional treatment. METHODS: The patients with neurotrophic keraitis were treated with 20% umbilical cord serum eyedrops 6 to 10 times a day. Ophthalmic examinations including best-corrected visual acuity (VA) measurement, corneal sensitivity test, corneal fluorescein staining, and anterior segment photography were performed before and after the treatment. Concentrations of substance P, insulinlike growth factor 1 (IGF-1), and nerve growth factor (NGF) in umbilical cord serum, normal peripheral blood serum, and tears were measured. MAIN OUTCOME MEASURES: Epithelial healing time; changes of VA and corneal sensitivity after treatment; and levels of substance P, IGF-1, and NGF in umbilical cord serum, normal peripheral blood serum, and tears. RESULTS: The epithelial defect healed completely in all eyes, with a mean healing time of 4.4+/-4.0 weeks. The epithelial defect healed within 2 weeks in 8 eyes (28.6%), between 2 and 4 weeks in 14 eyes (50.0%), and after 4 weeks in 6 eyes (21.4%). After treatment, VA improved by >2 lines in 17 eyes (60.7%). Mean pretreatment corneal sensitivity was 21.1+/-10.5 mm, and mean posttreatment corneal sensitivity was 24.3+/-11.7 mm (P<0.01). Mean concentrations of substance P, IGF-1, and NGF were 245.3+/-53.9 pg/ml, 239.0+/-77.1 ng/ml, and 729.7+/-72.0 pg/ml in umbilical cord serum; 169.5+/-81.0 pg/ml, 375.5+/-51.3 ng/ml, and 401.7+/-98.1 pg/ml in peripheral blood serum; and 69.8+/-24.9 pg/ml, 75.7+/-50.5 ng/ml, and 107.5+/-70.9 pg/ml in tears, respectively. CONCLUSIONS: Umbilical cord serum contains many neurotrophic factors, and umbilical cord serum eyedrops appeared to be effective for the treatment of neurotrophic keratitis.

Human leukocyte antigen B27 and B51 double-positive Behçet uveitis.

OBJECTIVES: To describe the clinical characteristics of human leukocyte antigen (HLA) B27 and B51 double-positive Behçet uveitis and to determine whether the coexistence of HLA-B27 can affect Behçet uveitis. METHODS: We retrospectively reviewed the medical records of patients with Behçet uveitis and patients with HLA-B27-associated non-Behçet uveitis who underwent HLA-B27 and HLA-B51 typing and were followed up for more than 3 years. We divided the patients into 3 groups according to HLA-B27/B51 status and compared the clinical outcomes. Main outcome measures were demographic features, uveitis characteristics, complications, treatments, and visual prognosis. RESULTS: Fourteen patients with HLA-B27(+)B51(+) Behçet uveitis, 43 patients with HLA-B27(-)B51(+) Behçet uveitis, and 41 patients with HLA-B27(+)B51(-) non-Behçet uveitis were identified. HLA-B27(+)B51(+) Behçet uveitis showed the demographic features similar to HLA-B27(-) counterparts. However, HLA-B27(+)B51(+) Behçet uveitis showed less involvement of posterior segments, a less chronic course, fewer complications in posterior segments, and less use of systemic medication or surgical intervention for inflammatory control, similar to HLA-B27(+)B51(-) non-Behçet uveitis. The long-term vision prognosis of HLA-B27(+)B51(+) Behçet uveitis was more favorable than that of HLA-B27(-)B51(+) Behçet uveitis. CONCLUSIONS: Our results indicate that HLA-B27(+)B51(+) Behçet uveitis is a benign subgroup of Behçet uveitis. The positivity of HLA-B27 may be a good prognostic factor in Behçet uveitis.

Comparison of autologous serum and umbilical cord serum eye drops for dry eye syndrome.

PURPOSE: To compare the therapeutic effect between autologous serum and umbilical cord serum eye drops in the treatment of severe dry eye syndrome. DESIGN: Prospective case-control study. METHODS: Ninety-two eyes of 48 patients with severe dry eye syndrome (34 eyes of 17 patients with Sjögren syndrome and 58 eyes of 31 patients with non-Sjögren syndrome) were treated with either 20% autologous serum (41 eyes of 21 patients) or umbilical cord serum eye drops (51 eyes of 27 patients). Symptom scoring, corneal sensitivity test, tear film break-up time (BUT), Schirmer test, tear clearance rate (TCR), corneal fluorescein staining, and conjunctival impression cytologic analysis were performed before and one month and two months after treatment. RESULTS: Both autologous serum and umbilical serum treatments led to improvement in the symptom score, tear film BUT, keratoepitheliopathy score, and impression cytologic findings. Symptom and keratoepitheliopathy scores were lower at one month (P = .03 and P = .12) and two months (P = .04 and .02) for those treated with umbilical cord serum compared with those treated with autologous serum. In Sjögren syndrome patients, goblet cell density was higher at two months of umbilical cord serum treatment compared with autologous serum treatment (P = .04). CONCLUSIONS: Umbilical cord serum eye drops were more effective in decreasing symptoms and keratoepitheliopathy in severe dry eye syndrome and increasing goblet cell density in Sjögren syndrome compared with autologous serum eye drops.

Photodynamic therapy with verteporfin for corneal neovascularization.

PURPOSE: To investigate the efficacy of photodynamic therapy with verteporfin for the treatment of patients with corneal neovascularization. DESIGN: Prospective interventional case series. METHODS: Eighteen eyes of 18 patients with stable corneal neovascularization who were refractory to conventional treatment were treated with photodynamic therapy with verteporfin (6 mg/m(2)). Five patients were treated following penetrating keratoplasty (PK), and two patients were treated before PK. Anterior segment photography was performed before and after treatment. Best-corrected visual acuity (BCVA) and area of corneal neovascularization were measured. RESULTS: At the one-year follow-up, 14 eyes (77.8%) showed a decrease in corneal neovascularization, and nine eyes (50.0%) showed complete vascular occlusion. In five patients who had corneal allograft, complete or partial occlusion was achieved in all eyes. Two patients who underwent subsequent keratoplasty did not manifest allograft rejection or revascularization. Seventeen eyes (94.4%) had stable or improved vision. The mean area of corneal neovascularization significantly decreased from 25.5 +/- 14.2 mm(2) to 14.9 +/- 14.6 mm(2) (P < .01), respectively. No significant complications associated with photodynamic therapy were observed except mild stromal haze in one eye. CONCLUSION: Photodynamic therapy with verteporfin may be effective for the treatment of corneal neovascularization.

Interleukin-6 and tumor necrosis factor-alpha levels in tears of patients with dry eye syndrome.

PURPOSE: To determine the levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in tears of patients with dry eye syndrome. METHODS: IL-6 and TNF-alpha levels were measured by enzyme-linked immunosorbent assay in tear samples obtained from 18 patients with dry eye (8 patients with Sjögren syndrome and 10 patients with non-Sjögren syndrome) and 14 control subjects. The correlation between IL-6 and TNF-alpha levels and tear film and ocular surface parameters was analyzed. The relative expression of these cytokines was evaluated in conjunctival impression cytology and conjunctival biopsy specimens by using immunohistochemical staining. RESULTS: The mean levels of IL-6 and TNF-alpha were, respectively, 18.57 +/- 8.92 and 3.68 +/- 3.45 pg/mL in patients with dry eye and 3.59 +/- 3.38 (P < 0.01) and < 0.5 (P < 0.01) pg/mL in control subjects. IL-6 level was significantly increased in tears of patients with Sjögren syndrome compared with those with non-Sjögren syndrome (P < 0.01). IL-6 level correlated significantly with tear film breakup time (P = 0.04), Schirmer test (P < 0.01), tear clearance (P = 0.02), keratoepithelioplasty score (P < 0.01), and goblet cell density (P = 0.03), but not with corneal sensitivity (P = 0.08). There was no significant difference in TNF-alpha level between patients with non-Sjögren and Sjögren syndrome. TNF-alpha levels did not correlate with tear film and ocular surface parameters. Immunohistochemical staining showed positive staining for IL-6 in specimens from patients with dry eye, especially in specimens from patients with Sjögren syndrome. CONCLUSION: IL-6 and TNF-alpha levels are elevated in tears of patients with dry eye syndrome. IL-6 level, but not TNF-alpha level, is associated with the severity of the disease and correlates with various tear film and ocular surface parameters.

An epidemic of chronic pseudophakic endophthalmitis due to Ochrobactrum anthropi: clinical findings and managements of nine consecutive cases.

PURPOSE: To report an epidemic of O. anthropi pseudophakic endophthalmitis. METHODS: The medical records of nine patients with culture-proven O. anthropi endophthalmitis were reviewed. RESULTS: The presenting features were compatible to chronic endophthalmitis. Two patients showed coinfections with P. acnes. Antibiotics sensitivity test revealed susceptibility to quinolones. Pars plana vitrectomy (PPV) with partial capsulectomy (PC) cured infections in seven patients without coinfection of P. acnes. Final visual acuity was 20/40 or better in five patients. CONCLUSIONS: O. anthropi should be considered in cases with chronic pseudophakic endophthalmitis. PPV with PC should be the initial therapeutic option for O. anthropi endophthalmitis.

Intraocular cytokine environment in active Behçet uveitis.

PURPOSE: To determine cytokine profiles in aqueous humor and peripheral blood from patients with Behçet uveitis. DESIGN: Prospective, experimental, and case-control study. METHODS: Aqueous humor and peripheral blood samples from 24 patients with Behçet uveitis, 28 patients with other causes of endogenous uveitis, and 20 healthy subjects after uncomplicated cataract surgery were obtained. Cytokines (interferon gamma (IFN-gamma), interleukin [IL]-2, IL-4, IL-10, IL-12, IL-15, and tumor necrosis factor alpha (TNF-alpha)) were measured by enzyme-linked immunosorbent assay and compared between the uveitis groups. RESULTS: Aqueous interferon gamma (IFN-gamma) levels were higher in patients with Behçet uveitis in contrast to higher concentrations of IL-4 in patients without Behçet uveitis. Aqueous levels of tumor necrosis factor alpha (TNF-alpha) were higher in patients with Behçet uveitis than in patients without Behçet uveitis. Aqueous and serum IL-10 were detected in all patients without Behçet uveitis, but were below detection limits in all patients with Behçet uveitis. Furthermore, IL-15 was exclusively increased in the aqueous humor of Behçet uveitis. The aqueous IL-2 or IL-12 levels were similar regardless of the presence of Behçet uveitis. CONCLUSIONS: Our results showed that an extreme Th1 polarization, a high proinflammatory condition, a low immunosuppressive status, and the presence of natural killer (NK) or CD8+ T cell-activating cytokine were unique features in aqueous humor with Behçet uveitis, suggesting that different immunopathogenic mechanisms may be involved in the ongoing intraocular inflammation of Behçet uveitis.

Application of umbilical cord serum eyedrops for the treatment of dry eye syndrome.

PURPOSE: To investigate the efficacy of umbilical cord serum eyedrops for the treatment of severe dry eye syndrome. METHODS: Fifty-five eyes of 31 patients with severe dry eye syndrome were treated with umbilical cord serum eyedrops. Symptom scoring, tear film break-up time (BUT), Schirmer test, corneal sensitivity test, and corneal fluorescein staining were performed before and 1 and 2 months after treatment, and conjunctival impression cytology was performed before and 2 months after treatment. The concentrations of epidermal growth factor (EGF), vitamin A, and transforming growth factor-beta (TGF-beta) in umbilical cord serum and normal peripheral blood serum were measured. RESULTS: Two months after treatment, significant improvement was observed in symptom score (from 3.07 +/- 0.54 to 0.96 +/- 0. 58), BUT (from 3.96 +/- 1.56 to 5.45 +/- 2.54 seconds), and keratoepitheliopathy score (from 4.87 +/- 3.22 to 1.71 +/- 1.84) (P < 0.01). There was no statistically significant change in Schirmer and corneal sensitivity tests. In impression cytology, the grade of squamous metaplasia (from 2.35 +/- 0.72 to 1.44 +/- 0.69) and goblet cell density (from 80.91 +/- 31.53 to 154.68 +/- 43.06 cell/mm) improved significantly (P < 0.01). The mean concentrations of EGF, TGF-beta, and vitamin A were 0.48 +/- 0.09, 57.14 +/- 18.98, and 230.85 +/- 13.39 ng/mL in umbilical cord serum and 0.14 +/- 0.03, 31.30 +/- 12.86, and 372.34 +/- 22.32 ng/mL in peripheral blood serum, respectively. CONCLUSION: Umbilical cord serum contains essential tear components, and umbilical cord serum eyedrops are effective and safe for the treatment of severe dry eye syndrome.

Experimental inhibition of corneal neovascularization by photodynamic therapy with verteporfin.

PURPOSE: To investigate the anti-angiogenic effects of photodynamic therapy with verteporfin in a rabbit model of corneal neovascularization. METHODS: One week after suturing, the localization of verteporfin in the neovascularized cornea was examined through fluorescent microscopy 1 hr after administration. Rabbits were treated with one or two times of photodynamic therapy with verteporfin at 1-week intervals. Analysis of corneal neovascularization was performed by biomicroscopic and histological examinations. RESULTS: Fluorescent microscopy showed green fluorescence in the vascular walls and interstitial tissue of the corneal stroma. The mean percentages of neovascularized corneal area at 3 days, 1 week, and 2 weeks after one time of photodynamic therapy were 90.3% +/- 3.5%, 71.6% +/- 6.2%, and 43.6% +/- 15.1% in treated eyes and 96.4% +/- 1.9% (p = 0.10), 88.6% +/- 4.6% (p = 0.01), and 76.8% +/- 4.4% (p < 0.01) in control eyes, respectively. The mean percentages 3 days, 1 week, and 2 weeks after two times of photodynamic therapy were also significantly lower in treated eyes compared with control eyes. In quantitative histological examination at 1 and 2 weeks after therapy, treated eyes showed significantly less neovascular area and number of vessels than control eyes. CONCLUSIONS: Photodynamic therapy with verteporfin is a safe and useful procedure to reduce experimental corneal neovascularization and can be used to inhibit angiogenesis in the cornea.

Combined low dose cyclosporine and prednisone down-regulate natural killer cell-like effector functions of CD8brightCD56+ T cells in patients with active Behçet uveitis.

PURPOSE: To investigate the changes of effector-related phenotypic markers and the natural killer (NK)-like effector functions of CD8(bright)CD56+ T cells in patients with Behçet uveitis after combined cyclosporine and prednisone (Cs/Pd) treatment. METHODS: Ten patients with active Behçet panuveitis and 10 healthy controls were prospectively recruited in this study. The effector-related surface markers (CD27, CD62L, CD11b, HLA-DR, CD94, NKG2D), chemokine receptors (CXCR1, CXCR3, CCR4, CCR5), and intracellular perforin of circulating CD8(bright)CD56+ T cells were determined by flow cytometric analysis before and after two months' treatment. NK-like cytotoxicity of ex vivo CD8(bright)CD56+ T cells against K562 was measured by standard 51Cr release assay. RESULTS: The expression levels of effector-related molecules on CD8(bright)CD56+ T cells normalized after treatment. The expression levels of CXCR1 and CCR5 were down-regulated on CD8(bright)CD56+ T cells after treatment. The amounts of preformed intracellular perforin of CD8(bright)CD56+ T cells were reduced to the normal levels. Furthermore, the NK-like cytolytic capacities of CD8(bright)CD56+ T cells were decreased after treatment. CONCLUSIONS: Our results suggest that the combined Cs/Pd treatments in active Behçet uveitis may downregulate the NK-like effector functions of CD8(bright)CD56+ T cells.

Therapeutic effect of umbilical cord serum eyedrops for persistent corneal epithelial defect.

PURPOSE: To evaluate the therapeutic effect of umbilical cord serum in the treatment of persistent epithelial defect of the cornea. METHODS: Fourteen eyes of 14 patients with persistent epithelial defect that had persisted for at least 2 weeks despite conventional treatment were treated with 20% umbilical cord serum eyedrops six times a day. The images of the epithelial defects were captured using a camera attached to a slit lamp biomicroscope and the areas of the epithelial defects were calculated. Treatment was considered effective for epithelial defect healing within 2 weeks, partially effective for healing within 2 to 4 weeks, and ineffective for healing requiring either more than 1 month or additional measures. RESULTS: Mean duration of epithelial defect before treatment was 7.2 +/- 6.3 weeks, and mean area was 7.86 +/- 7.32 mm2. Umbilical cord serum therapy was effective in 6 eyes (42.9%), partially effective in 6 (42.9%), and ineffective in 2 (14.2%). Nevertheless, the epithelial defects in both the ineffective eyes were eventually healed within 8 weeks. Mean healing time in effective or partially effective cases was 2.75 +/- 1.06 weeks. CONCLUSIONS: The use of umbilical cord serum eyedrops for the treatment of persistent epithelial defect is effective.

Eyelid sparganosis.

PURPOSE: To report a case of sparganosis in the muscle layer of the eyelid. DESIGN: Observational case report. METHODS: A 67-year-old man with migratory painful swelling on the eyelid that was unresponsive to medications was evaluated and treated surgically. RESULTS: Computed tomography showed a 1.5 x 1.5-cm sized, thick-walled lesion and ultrasound biomicroscopy showed hypoechoic tubular and cystic lesions. During surgery, a white, thread-like plerocercoid, 7 cm in length was detected in the orbicularis muscle of the lower eyelid. Histopathologic examination demonstrated the characteristic feature of the sparganum larva and foreign body granulomatous reaction. Serodiagnosis by enzyme-linked immunosorbent assay was positive. Three months postoperatively, the lesion resolved completely. CONCLUSION: Although rare, sparganosis should be suspected in a moving eyelid mass unresponsive to the medical treatment.

Recurrent Salzmann's nodular degeneration.

BACKGROUND: We report a patient with Salzmann's nodular degeneration in whom the disease occurred without a history of keratitis and recurred twice after keratectomy. CASE: A unilateral case of symptomatic Salzmann's nodular degeneration developed after cataract surgery in a 75-year-old woman. OBSERVATIONS: We performed superficial keratectomy and amniotic membrane transplantation two times, but both times the lesions recurred 1 month postoperatively. After we performed the third superficial keratectomy above the level of Bowman's layer, there was no recurrence by the last follow-up 6 months postoperatively. CONCLUSIONS: Salzmann's nodular degeneration can be diagnosed by clinical characteristics, even if there is no history of keratitis. Because it can recur after surgical removal of the lesion, patients with this disease need to be followed-up periodically.

Mechanical treatment of phthiriasis palpebrarum.

Phthiriasis palpebrarum is a rare disease in which crab lice infest the eyelashes. It can cause pruritic lid margins or unusual blepharoconjunctivitis and is difficult to diagnose and treat. We diagnosed and managed a case of phthiriasis palpebrarum in both upper eyelids, accompanied by nits, on the scalp of a 6 year-old female child. We removed the eyelashes, including lice and nits, by pulling with fine forceps without sedation. On the second month after treatment, all lice and nits were eradicated without recurrence and the eyelashes grew back. In conclusion, phthiriasis palpebrarum can be diagnosed by close examination of the eyelashes and eyelid margins with slit lamp and can be managed mechanically.