A novel gene mutation in a family with X-linked retinoschisis.

BACKGROUND/PURPOSE: To describe the clinical characteristics of a Taiwanese family with X-linked retinoschisis (XLRS) and to investigate the molecular genetics of a novel mutation in the retinoschisin 1 (RS1) gene. METHODS: A total of 15 participants in this XLRS family were analyzed. Complete ophthalmic examinations and fundus photography were performed on 15 family members. These tests identified five affected males and two female carriers. Blood samples were collected, and genomic DNA was extracted. Best-corrected visual acuity, optical coherence tomography (OCT), electroretinogram (ERG), and direct DNA sequence analysis of the RS1 gene were performed on 15 family members. RESULTS: Five affected males, with visual acuity ranging from 0.2 to 0.7, had macular schisis and abnormal retinal pigment epithelium pigmentation. The mixed scotopic ERG "b" wave was more reduced than the "a" wave. OCT revealed typical microcystic schisis cavities within the macula area. Direct DNA sequence analysis revealed a single base pair deletion, 97delT, in all the affected individuals. This deletion resulted in a frameshift mutation of the RS1 gene, causing protein truncation. The affected males in this family showed moderately decreased visual acuity and dysfunction in both cone cells and phototransduction. CONCLUSION: We identified a novel RS1 (97delT) mutation in a Taiwanese family with XLRS. This finding expands the RS1 mutation spectrum and may help to further understand the molecular pathogenesis of XLRS.

The oculocardiac reflex during strabismus surgery: its relationship to preoperative clinical eye findings and subsequent postoperative emesis.

PURPOSE: To compare the occurrence of significant bradycardia due to the oculocardiac reflex (OCR) during strabismus surgery and its relationship to preoperative clinical eye findings and subsequent development of postoperative emesis. METHODS: The medical records of consecutive patients who underwent strabismus surgery August 2006 to June 2009 were retrospectively reviewed. OCR was defined as presence of dysrhythmia or a sudden heart rate decrease ≥ 20% after traction on the extraocular muscle. OCR incidence was compared between the first, second, and third (if any) extraocular muscles in patients who had multiple-muscle strabismus surgery and also between specific muscles (eg, medial rectus vs lateral rectus muscle). Associations with OCR were compared for different strabismus types. Vomiting was considered postoperative if it occurred before discharge of the patient at noon on the day following surgery. Risk factors for OCR and postoperative vomiting were evaluated by logistic regression analysis. RESULTS: A total of 111 records were reviewed; 41 patients (37%) experienced OCR. Incidence of OCR and absence of OCR during traction of the first muscle were significantly associated with events during traction of the second (χ(2) = 36.681, P < 0.001) and third muscles (Fisher exact test, P = 0.030). The best predictors of OCR were the absence of fine stereopsis and a larger number of surgically treated muscles. Of the 111 patients, 30 (27%) who had postoperative vomiting, the best predictors were female sex and young age. CONCLUSIONS: In our cohort, patients experiencing intraoperative OCR when the first extraocular muscle was manipulated during multiple-muscle strabismus surgery were likely to experience it again during traction of additional muscles. More severe postoperative vomiting was common in these patients. OCR was associated with the preoperative absence of fine stereopsis.

Uncorrected visual acuity and noncycloplegic autorefraction predict significant refractive errors in Taiwanese preschool children.

PURPOSE: To investigate the accuracy of uncorrected visual acuity (UCVA), stereopsis, and noncycloplegic autorefraction (NCAR) tests performed by vision-screening technicians and to determine the best referral criteria when using these methods to screen for significant refractive errors in preschool children. DESIGN: Retrospective, case-control, and cross-sectional study. PARTICIPANTS: We reviewed 1000 records for a population-based preschool vision-screening program. The target conditions were defined as myopia ≤-3.0 diopters (D), hyperopia ≥ 4.5 D, astigmatism ≥ 2.0 D, and anisometropia ≥ 2.0 D. METHODS: Receiver operating characteristic (ROC) curve was used to calculate optimal referral cutoff values. The examination results obtained by the vision-screening technicians were compared with those obtained by a pediatric ophthalmologist, which were considered the gold standard. MAIN OUTCOME MEASURES: The efficacies (sensitivity, specificity, positive predictive value, and negative predictive value) of different tests were evaluated. RESULTS: In 7.0% (95% confidence interval [CI], 5.3-8.7) of the children, at least 1 eye showed 1 of the target conditions. If only the right eyes were considered, the prevalence of target conditions was 4.2% (95% CI, 2.9-5.5). The ROC curve analysis indicated that the NCAR cylinder test (cutoff value ≥ 0.875 D) was the best test for screening target conditions. With regard to age groups, UCVA ≤ 0.75 (Snellen equivalent) and ≤ 0.85 were the best referral criteria for ages ≤ 4 years and ≥ 5 years, respectively. Combining the UCVA test with the NCAR test (the child was referred after failing both tests) increased specificity without significantly decreasing sensitivity. CONCLUSIONS: The UCVA and NCAR tests performed by vision-screening technicians are adequately sensitive and specific for preschool vision screening. The ROC curve analysis was used for determining the appropriate screening criteria for these tests, and combining the tests increased their accuracy. The screening criteria should be age dependent. When analyzing the test accuracy in ophthalmic problems, if the disease of interest does not symmetrically (in terms of disease severity and prevalence) involve both eyes, the prevalence based on only 1 eye should be interpreted with caution.

Extraocular muscle insertion positions and outcomes of strabismus surgery: correlation analysis and anatomical comparison of Western and Chinese populations.

BACKGROUND/AIMS: To compare the insertion locations of extraocular muscles between Taiwanese (Han Chinese) and Western populations and to determine whether anatomical differences warrant different surgical guidelines. METHODS: Insertion locations were compared between a Taiwanese population of subjects who had received surgical treatment for strabismus and a control group who had not. Insertion locations and surgical outcomes in the strabismus group were also compared with those reported in other countries. RESULTS: In Taiwanese subjects, extraocular muscle insertion locations were not significantly different between strabismus subjects and controls. However, the distances from the insertion location to the limbus of the inferior rectus, lateral rectus and superior rectus were significantly shorter in the Taiwanese subjects than in Western populations. CONCLUSION: Extraocular muscle insertion locations for the Taiwanese population in this study significantly differed from those reported in studies of Western populations. Therefore, surgical guidelines for performing lateral rectus recession to treat exotropia in Western populations may be inappropriate for Taiwanese and other Asian populations.

Development of visual acuity in preschool children as measured with Landolt C and Tumbling E charts.

PURPOSE: To investigate the visual acuity development as measured by Landolt C and Tumbling E charts ("C chart" and "E chart") in preschool children. METHODS: Visual acuity measurements of healthy children ages 3-6 years who had been evaluated with the use of E charts (during kindergarten screening) and C charts (in a hospital setting) from 2005 to 2007 were studied. Inclusion criteria were hyperopia <4.5 D, myopia <3.0 D, astigmatism <2.0 D, and anisometropia <2.0 D. The best-corrected visual acuity (BCVA) in each group was analyzed. RESULTS: There were 821 children in the E chart group and 212 in the C chart group (mean age, 5 years). The mean BCVA improved from between age 3-4 years and age 5-6 years in both groups (P < 0.001). Children in the C chart group had significantly greater astigmatism compared with those in the E chart group (0.66 D ± 0.47 vs 0.28 D ± 0.39; P < 0.001). The BCVA obtained by E chart was greater than that obtained by C chart at age 3-4 years (logMAR 0.0246 ± 0.0646 vs 0.1158 ± 0.1462, P < 0.001), but not at age 5-6 years. CONCLUSIONS: When children reach 5-6 years of age, visual acuity values obtained by C and E charts approach those observed in adults. In younger patients, the E chart obtains a greater visual acuity score than the C chart. Thus, thresholds for normal visual acuity in preschool children ages 3-6 years should be both age specific and test specific.

Astigmatism in preschool children in Taiwan.

PURPOSE: To elucidate the prevalence of astigmatism and its corneal component, the association between the amount of astigmatism and its axis, and the association between the axis of astigmatism and body mass index (BMI) in Taiwanese preschool children. METHODS: In this population-based study we analyzed 2 data sets, one obtained by retrospective review of vision-screening data and another obtained in a prospective observational study. Each study collected data for age, sex, and refraction status. Autokeratometry, height, and weight measurements were obtained only in the observational study. Astigmatism was classified as with-the-rule, against-the-rule, or oblique. RESULTS: The prevalence of astigmatism was unassociated with age or sex in both data sets. The retrospective study (1,094 subjects; mean age, 5 years; range, 2.19-7.32 years) revealed that 13.3% of the subjects had astigmatism > or = 1.00 D, and most of them had with-the-rule astigmatism; 4.0% had high astigmatism (>1.50 D). Children with with-the-rule astigmatism had greater mean cylinder power than those with against-the-rule or oblique astigmatism. In the prospective study (190 subjects; mean age, 5.65 years; range, 3.79-6.68 years) astigmatism correlated with its corneal component. Preschoolers with with-the-rule astigmatism and with-the-rule corneal astigmatism had greater BMI and heavier weight than those with against-the-rule or oblique type. CONCLUSIONS: Astigmatism is common in Taiwanese preschool children. The predominant types are with-the-rule and oblique astigmatism. Most patients with high astigmatism and high corneal astigmatism have with-the-rule type. High BMI or weight is associated with with-the-rule astigmatism and with with-the-rule corneal astigmatism.

Mechanism of carteolol-induced cytosolic Ca2+ mobilization in cultured vascular smooth muscle cells.

An increase in cytosolic calcium concentration triggers intracellular signal transduction in vascular cells, which then regulates the vascular contraction. In the present study, the regulatory mechanism of carteolol on the intracellular free Ca(2+) ([Ca(2+)](i)) mobilization was investigated in cultured A7r5 vascular smooth muscle cells. The A7r5 cells were cultured and loaded with fura-2-AM, which was used as a Ca(2+) sensitive fluorescent probe. In both the presence and absence of external Ca(2+), carteolol increased [Ca(2+)](i) with a dose-dependent manner in A7r5 cells at concentrations between 608 microM and 6.08 microM. In a Ca(2+)-containing buffer, carteolol-induced [Ca(2+)](i) showed an initial peak followed by a secondary and persistent plateau. Pretreatment of the cells with La(3+), the plasma membrane Ca(2+) pump inhibitor, and nifedipine, a L-type Ca(2+) channel inhibitor, both partially restrained the carteolol-induced initial peak in [Ca(2+)](i) by 92% and 86%, respectively. Pretreatment of the cells with adrenoceptor antagonists, prazosin inhibited the [Ca(2+)](i) response by 80%, and propranolol enhanced the response by 61%. In the Ca(2+-)-free buffer, pretreatment of the cells with carteolol inhibited the endoplasmic reticulum Ca(2+) pump inhibitor of thapsigargin-induced [Ca(2+)](i) increase by 97%. Pretreatment of the cells with thapsigargin also inhibited the carteolol-induced [Ca(2+)](i) rise by 98%. The internal Ca(2+) release induced by the carteolol was partially inhibited by U73122 (phospholipase C inhibitor) and aristolochic acid, quinacrine (phospholipase A(2) inhibitors). After incubation of carteolol in the Ca(2+)-free buffer, the addition of CaCl(2) increased the Ca(2+) influx, implying that the release of Ca(2+) from internal stores further induced capacitative Ca(2+) entry. These results suggest that carteolol-induced [Ca(2+)](i) increase is mediated by the initial influx via the alpha(1)-adrenoceptor, L-type Ca(2+) channel, nonselective calcium entry channels and release of Ca(2+) from an intracellular store, which is mainly in the endoplasmic reticulum followed by capacitative Ca(2+) entry but decrease via the beta(2)-adrenoceptor. The intracellular Ca(2+) release was also modulated by phospholipase A(2), C-coupled events.

The visual status of children ages 3 to 6 years in the vision screening program in Taiwan.

PURPOSE: To investigate the current status of myopia, amblyopia, and strabismus of preschoolers in Taiwan. METHODS: The records of a vision screening performed in 2005 at 4 preschools by a tertiary referral medical center in Kaohsiung, Taiwan were reviewed. Cycloplegic refraction, visual acuity by E chart, stereopsis by Titmus and National Taiwan University tests, and ocular alignment were analyzed. RESULTS: Most (97.7%) of the children (n = 618) were able to perform visual acuity testing. There was good correlation between the results of cycloplegic spherical equivalent obtained by autorefraction and by retinoscopy (r = 0.934, p < 0.001). The prevalence of amblyopia was about 5%. A trend was found between amblyopia morbidity and the 4 age groups when a single criterion (best-corrected visual acuity <0.7) was used for all ages (decreased amblyopia incidence with increased age; p = 0.028, chi(2) test for trend). The prevalence of myopia was 3.0%, 4.2%, 4.7%, and 12.2% at ages 3, 4, 5, and 6 years, respectively. There was a trend of increased myopia with increased age (p = 0.021, chi(2) test for trend). CONCLUSIONS: The prevalence of myopia in children ages 3 to 6 years in Taiwan was high compared with the prevalence in other countries. Criteria for amblyopia based on age difference and on the types of visual acuity test are recommended.

Ethambutol induces PKC-dependent cytotoxic and antiproliferative effects on human retinal pigment cells.

Ethambutol (EMB)-induced ocular side effects may involve the influence on functions of retinal pigment epithelium (RPE), in addition to EMB-induced optic neuropathy. To address this issue, the molecular and cellular effects of EMB on RPE including growth regulation, morphological responses, phagocytic activity, and the relevant signaling pathways were investigated. EMB (at optimal concentration 8.0mM) can trigger cell cycle arrest in both RPE50 and ARPE19 cells, accompanied by reduced DNA synthesis. EMB also induced cytoplasmic vacuole formation in both RPE cell lines. Under transmission electric microscope, the phagosomes were replaced by vacuoles and the number of microvilli was reduced in EMB-treated cells. Animal experiments also demonstrated the vacuole formation within RPE of the EMB-treated rats. On the other hand, by in vitro phagocytosis assay using rod outer segment (ROS) as the target, we found EMB suppressed phagocytosis in the cultured RPE, which is consistent with the decreased rhodopsin uptake in the RPE of the EMB-treated rats. Furthermore, inhibitor of protein kinase C but not MAPK, prevented the EMB-induced phenotypical changes. Using a non-radioactive PKC assay, we also demonstrated the PKC activity in both RPE cell lines can be induced by EMB. In conclusion, EMB may exert toxic effects in RPE including suppression of cell growth, formation of cytoplasmic vacuoles and reduction of phagocytic functions via PKC signal pathway.

Mechanism of mitomycin-induced apoptosis in cultured corneal endothelial cells.

PURPOSE: Previous studies have indicated that improper use of mitomycin C (MMC) caused cytotoxicity in corneal endothelial cells. The aim of the present study was to investigate whether MMC induces cellular apoptosis in corneal endothelial cells and to determine the mechanism by which this may occur. METHODS: Porcine corneal endothelial cells were acquired from primary culture. Cellular damage and caspase pathway were estimated with a MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. The apoptotic characteristics were detected by means of flow cytometry, the TUNEL (terminal deoxyribonucleotidyl-transferase-(TdT)-mediated deoxyuridine-5'-triphosphate-digoxigenin (dUTP) nick-end labeling) test, immunofluorescent staining, and western blotting. RESULTS: The results indicated that MMC was toxic to corneal endothelial cells in a time-dependent and dose-dependent manner. Pretreatment with a general caspase inhibitor (Z-VAD-FMK), a caspase-8 inhibitor (Z-IETD-FMK), and a caspase-9 inhibitor (Z-LEHD-FMK) reversed MMC-induced cellular damage. Following exposure to MMC, a change in the mitochondrial membrane potential was positively detected by flow cytometric assay with MitoLight dye while cellular cytochrome c that was released from the mitochondria to the cytoplasm was detected by immunofluorescent staining. A positive TUNEL test revealed that cellular DNA apoptosis had occurred following exposure to 0.001 and 0.01 mg/ml MMC for 24 h. Positive annexin V-FITC, and negative propidium iodide (PI) staining indicated that the cellular plasma membrane underwent apoptosis following 0.001 mg/ml MMC exposure for 24 h. Western blot assay demonstrated down-regulation of the Bcl-2 protein and upregulation of the p53 and p21 proteins, which were all involved in apoptosis induced by MMC. CONCLUSIONS: These results indicate that mitomycin-induced cellular apoptosis in corneal endothelial cells may be mediated through caspase-8, caspase-9, and the mitochondrial regulated pathways as well as through upregulation of p53-dependent and p21-dependent signal transduction pathways.

Neuroprotective effects of recombinant human granulocyte colony-stimulating factor (G-CSF) in neurodegeneration after optic nerve crush in rats.

The purpose of the present study was to investigate the effects of granulocyte colony-stimulating factor (G-CSF) on neurodegeneration of optic nerve (ON) and retinal ganglion cells (RGCs) in a rat model of ON crush. The ONs of adult male Wistar rats (150-180 g) were crushed by a standardized method. The control eyes received a sham operation. G-CSF (100 microg/kg/day in 0.2 ml phosphate-buffered saline) or phosphate-buffered saline (PBS control) was immediately administered after ON crush for 5 days by subcutaneous injection. Rats were euthanized at 1 or 2 weeks after the crush injury. RGC density was counted by retrograde labeling with FluoroGold application to the superior colliculus, and visual function was assessed by flash visual evoked potentials (FVEP). TUNEL assay, Western blot analysis and immunohistochemistry of p-AKT in the retina and ED1 (marker of macrophage/microglia) in the ON were conducted. 2 weeks after the insult, the RGC densities in the central and mid-peripheral retinas in ON-crushed, G-CSF-treated rats were significantly higher than that of the corresponding ON-crushed, PBS-treated rats (survival rate was 60% vs. 19.6% in the central retina; 46.5% vs. 23.9% in mid-peripheral retina, respectively; p<0.001). FVEP measurements showed a significantly better preserved latency of the p1 wave in the ON-crushed, G-CSF-treated rats than the ON-crushed, PBS-treated rats (78+/-9 ms in the sham operation group, 98+/-16 ms in the G-CSF-treated group, and 174+/-16 ms in the PBS-treated group; p<0.001). TUNEL assays showed fewer apoptotic cells in the retinal sections in the ON-crushed, G-CSF-treated rats. p-AKT immunoreactivity was up-regulated in the retinas of the ON-crushed, G-CSF-treated rats at 1 and 2 weeks. In addition, the number of ED1-positive cells was attenuated at the lesion site of the optic nerve in the ON-crushed, G-CSF-treated group. From these results, we gather that administration of G-CSF is neuroprotective in the rat model of optic nerve crush, as demonstrated both structurally by RGC density and functionally by FVEP. G-CSF may work by being anti-apoptotic involving the p-AKT signaling pathway as well as by attenuation of the inflammatory responses at the injury site, as evidenced by less ED1-positive cell infiltration in the optic nerve.

Endothelin-1 enhances corneal fibronectin deposition and promotes corneal epithelial wound healing after photorefractive keratectomy in rabbits.

The objective was to study the effects of endothelin-1 (ET1) on corneal wound healing after photorefractive keratectomy (PRK) in rabbit corneas. Following PRK, 18 New Zealand white rabbits were treated with ET1 in the right eyes and with phosphate-buffered salt solution (PBS) in the left eyes. Corneal epithelial wound size, corneal haze and corneal thickness were recorded. Corneal extracellular matrixes, including collagen types 3, 4 and 7, chondroitin sulfate and fibronectin, were investigated using immunohistochemistry study. ET1 increased the rate of healing of corneal epithelial wounds in rabbits. Anti-fibronectin fluorescence was present at week 12 and week 24 in ET1-treated eyes but not in the control eyes. There were no significant differences in corneal haze, corneal thickness and changes in other extracellular matrixes between ET1- and PBS-treated eyes. ET1 can enhance the deposition of fibronectin in corneal stroma and promote corneal epithelial wound healing after PRK. The increase in fibronectin probably explains the increased healing rate of corneal epithelial wounds.

Mechanism of unoprostone-induced cytosolic Ca2+ mobility in cultured porcine corneal endothelial cells.

The effect of unoprostone isopropyl on the intracellular free Ca(2+) signaling ([Ca(2+)](i)) in cultured porcine corneal endothelial cells was evaluated by using fura-2-AM as a Ca(2+) probe. In Ca(2+)-containing buffer and Ca(2+)-free buffer, unoprostone increased [Ca(2+)](i) concentration dependently from 28.2 to 0.282microM, diluted from original concentration to 1/100, 1/1000 and 1/10,000. The response was inhibited on extracellular Ca(2+) removal. In Ca(2+)-free buffer, pretreatment of the cells with unoprostone inhibited the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin-induced [Ca(2+)](i) increase and the mitochondria uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP)-induced Ca(2+) release by 96 and 95%, respectively. Pretreatment of the cells with thapsigargin or CCCP also inhibited unoprostone-1-induced [Ca(2+)](i) rise by 84 and 57%, respectively. The intracellular calcium release caused by unoprostone was partially inhibited by phospholipase C inhibitor (U73122) and by phospholipase A(2) inhibitor aristolochic acid. After incubation of the cells with unoprostone in Ca(2+)-free buffer, the addition of 5mM CaCl(2) increased Ca(2+) influx, implying that release of Ca(2+) from internal stores caused by unoprostone further induced capacitative Ca(2+) entry. These results suggest that unoprostone-induced [Ca(2+)](i) increase in corneal endothelial cells results from Ca(2+) influx from external buffer and release of Ca(2+) from intracellular stores from the endoplasmic reticulum and mitochondria Ca(2+) stores followed by capacitative Ca(2+) entry. Unoprostone-induced intracellular Ca(2+) release was also modulated by phospholipase A(2) and C-coupled events.

Cellular cytotoxicity of antiglaucoma drugs in cultured corneal endothelial cells.

In this study, the various antiglaucoma drugs including betaxolol, timolol, levobunolol, carteolol, brimonidine, dipivefrin, dorzolamide, brinzolamide, latanoprost, unoprostone, and pilocarpine were used to investigate the effects of cellular cytotoxicity in cultured bovine corneal endothelial cells. After exposure to the drugs in three dilutions, 1/100, 1/1,000, and 1/10,000, for 100 minutes, cells were estimated based on the release assay of lactate dehydrogenase (LDH) enzyme. It was found that cellular LDH was significantly released in the medium only at 1/100th dilution of betaxolol, brimonidine, dorzolamide, dipivefrin, latanoprost and unoprostone to 130%, 123%, 145%, 157%, 128% and 237%, respectively, compared with controls upon exposure to drugs for 100 minutes. Moreover, benzalkonium chloride preservative at the concentrations ranging from 0.001 to 0.00001 mg/mL did not affect cellular LDH release in bovine corneal endothelial cells. These results indicate that high concentrations of antiglaucoma drugs may induce cytotoxicity in corneal endothelial cells.

Occult plastic intravitreal foreign body retained for 30 years: a case report.

A 41-year-old man had suffered trauma to the right eye 30 years ago. In March 1996, he underwent trabeculectomy and peripheral iridectomy under the diagnosis of open angle glaucoma in the right eye. Autoperimetry at that time revealed visual field constriction. In addition, ocular examination showed that the cup/disc ratio of his right eye was increased. Cataract was diagnosed in September 2002 and cataract extraction was performed on October 22, 2002. Aplastic intravitreal foreign body was detected during the operation. However, preoperative B-scan ultrasonography had failed to detect an intraocular foreign body (IOFB), and the previous fluorescein angiography had shown only retinal pigment epithelium changes. This case reminded us that we should be alert to an occult IOFB in the event of ocular trauma, even if none had been detected during prior imaging examinations.

Effects of antibiotics and corticosteroid eyedrops on cellular proliferation in cultured human corneal keratocytes.

The purpose of this study is to investigate the effects of antibiotics and glucocorticoid eyedrops, including gentamicin, sulfisomezole, fluorometholone, dexamethasone, and betamethasone, on cellular proliferation in cultured human corneal keratocytes. Human corneal keratocytes were cultured in RPMI-1640 containing 10% fetal bovine serum. Drugs were prepared from original concentrations to 1/10, 1/100, and 1/1,000 dilutions. After exposure to drugs for 100 minutes, cellular proliferation was estimated by [3H]-thymidine uptake. It was found that cellular proliferation in corneal keratocytes was not affected by any of the three dilutions of gentamicin but was inhibited by 1/10 and 1/100 dilutions of sulfisomezole to 82% and 90% of control. [3H]-thymidine uptake values were inhibited to 75% by 1/10 dilution of fluorometholone and by 1/10and 1/100 dilutions of betamethasone to 84% and 86% of control. Meanwhile, cellular proliferation was significantly inhibited by 1/10, 1/100, and 1/1,000 dilutions of dexamethasone to 82%,86%, and 90%, respectively, in comparison with control values. It was demonstrated that commercial eyedrops of glucocorticoids inhibit cellular proliferation in corneal keratocytes, which may modulate the wound healing of corneal stroma.

Effects of antiglaucoma drugs on cellular proliferation in cultured human corneal keratocytes.

The purpose of this study was to investigate the effects of various antiglaucoma drugs, including timolol, betaxolol, carteolol, levobunolol, dipivefrin, and pilocarpine, on cellular proliferation in cultured human corneal keratocytes. Human corneal keratocytes were cultured with RPMI-1640 medium containing 10% fetal bovine serum. Antiglaucoma drugs were prepared from original concentrations to dilutions of 1/10, 1/100, and 1/1,000. After exposure to drugs for 100 minutes, cellular proliferation was estimated by [3H]thymidine uptake methodology. It was found that cellular proliferation in corneal keratocytes was inhibited by only a 1/10 dilution of various drugs including timolol, betaxolol, carteolol, levobunolol, dipivefrin, and pilocarpine. The [3H]thymidine uptake values were significantly inhibited to 63%, 18%, 87%, 68%, 55%, and 67% by a 1/10 dilution of the above drugs. However, the cellular proliferation was also significantly suppressed by 0.01 mg/mL of benzalkonium chloride preservative. It is shown that the inhibition of cellular proliferation by high concentrations of antiglaucoma drugs may result from the benzalkonium chloride preservative contained in these drugs.

Polymerase chain reaction identification of a hymenopteran insect in the cornea: a case report.

The type of corneal injuries associated with insect encounters is related to the composition of the foreign body. However, previous reports on corneal foreign bodies as insects were rarely based on scientific evidence. Here, we report on a 49-year-old male who was stung in his left eye by an unknown insect. Emergent keratotomy was performed to remove the embedded corneal foreign body. The removed foreign body was observed under light microscopy, and a fragment of insect was suspected. The sample was sent for molecular analysis. The polymerase chain reaction product was sequenced, subjected to a BLAST search, and identified as an ichneumonoid member of the insect order Hymenoptera.