Cytochrome P450 monooxygenase of Acanthamoeba castellanii participates in resistance to polyhexamethylene biguanide treatment.

Acanthamoeba spp. are free-living parasites that can cause severe infections such as granulomatous amoebic encephalitis (GAE) and amoebic keratitis (AK). Polyhexamethylene biguanide (PHMB) is a topical application for AK treatment. However, PHMB is not entirely effective against all Acanthamoeba strains or isolates. The mechanisms by which Acanthamoeba protects itself against extreme drug conditions without encystation are still unknown. According to a previous study, cytochrome P450 monooxygenase (CYP450MO) plays an important role in the oxidative biotransformation of numerous drugs related to metabolism. In this study, a CYP450MO fragment was inserted into the pGAPDH-EGFP vector and transfected into Acanthamoeba castellanii. We found that CYP450MO-overexpressing Acanthamoeba had higher survival rates than those of the control cells after PHMB treatment. Moreover, we also found that encystation-related genes such as cellulose synthase I (CSI), encystation-mediating serine proteinase (EMSP), and autophagy-related protein 8 (ATG8) expression levels were not significantly different between Acanthamoeba transfected by pGAPDH-EGFP or pGAPDH-EGFP-CYP450MO. We suggest that Acanthamoeba transfected by pGAPDH-EGFP-CYP450MO may not induce encystation-related genes to resist PHMB treatment. In conclusion, these findings indicate that CYP450MO may be an additional target when PHMB is used for treatment of amoebic keratitis.

TITLE: La monooxygénase du cytochrome P450 d'Acanthamoeba castellanii participe à la résistance au traitement par le polyhexaméthylène biguanide. ABSTRACT: Les Acanthamoeba spp. sont des parasites libres qui peuvent provoquer des infections graves telles que l'encéphalite amibienne granulomateuse (EAG) et la kératite amibienne (KA). Le polyhexaméthylène biguanide (PHMB) est une application topique pour le traitement de la KA. Cependant, le PHMB n'est pas entièrement efficace contre toutes les souches ou isolats d'Acanthamoeba. Les mécanismes par lesquels Acanthamoeba se protège contre des conditions médicamenteuses extrêmes sans enkystation sont encore inconnus. Selon une étude précédente, la monooxygénase du cytochrome P450 (CYP450MO) joue un rôle important dans la biotransformation oxydative de nombreux médicaments liés au métabolisme. Dans cette étude, un fragment CYP450MO a été inséré dans le vecteur pGAPDH-EGFP et transfecté dans Acanthamoeba castellanii. Nous avons constaté que les Acanthamoeba surexprimant le CYP450MO avaient des taux de survie plus élevés que ceux des cellules témoins après un traitement au PHMB. De plus, nous avons également constaté que les gènes liés aux enkystations tels que la cellulose synthase I (CSI), la sérine protéinase médiatrice de l'enkystation (EMSP) et les niveaux d'expression de la protéine 8 liée à l'autophagie (ATG8) n'étaient pas significativement différents entre les Acanthamoeba transfectés par pGAPDH-EGFP ou par pGAPDH-EGFP-CYP450MO. Nous suggérons que les Acanthamoeba transfectés par pGAPDH-EGFP-CYP450MO ne peuvent pas induire les gènes liés à l'enkystation pour résister au traitement PHMB. En conclusion, ces résultats peuvent indiquer que la monooxygénase du cytochrome P450 peut être une cible potentielle pour le traitement par PHMB de la kératite amibienne.

Ocular characteristics in a variant microcephalic primordial dwarfism type II.

BACKGROUND: Microcephalic osteodysplastic primordial dwarfism, type II (MOPD II) is a rare disease that is assumed to be caused by a pericentrin (PCNT) gene mutation. Clinical manifestations have been reported in pediatrics and neurology; however, only a few ocular findings have been documented. CASE PRESENTATION: We present three unrelated cases of MOPD II with similar facial features and short stature. Unlike the cases described in the literature, all subjects had normal birth weight and height but their growth was retarded thereafter. In addition to delayed milestones, they have a broad forehead, maxillary protrusion, long peaked nose, high nasal bridge, low-set large ears, extreme reromicrogenia, and normal-sized teeth. These three patients had similar ocular manifestations with the short axial length associated with high hyperopia more than + 9 diopters (D) and macular scarring. The oldest subject was a 20 year-old male without neurological symptoms. One female subject had developed alopecia during the previous 2 years. The other female subject had moyamoya disease, but a genetic study revealed a normal PCNT gene. CONCLUSION: This is the first report of MOPD II focusing on ocular findings, suggesting that macular dystrophy and high hyperopia are the common ocular characteristics of MOPD II. Prompt referral to an ophthalmologist is essential. Although refractive amblyopia can be treated with optical correction, visual prognosis may be poor due to maculopathy.

Identification and characterization of a secreted M28 aminopeptidase protein in Acanthamoeba.

Acanthamoeba is a free-living pathogenic protozoan that is distributed in different environmental reservoirs, including lakes and soil. Pathogenic Acanthamoeba can cause severe human diseases, such as blinding keratitis and granulomatous encephalitis. Therefore, it is important to understand the pathogenic relationship between humans and Acanthamoeba. By comparison of systemic analysis results for Acanthamoeba isolates, we identified a novel secreted protein of Acanthamoeba, an M28 aminopeptidase (M28AP), which targets of the human innate immune defense. We investigated the molecular functions and characteristics of the M28AP protein by anti-M28 antibodies and a M28AP mutant strain generated by the CRISPR/Cas9 system. Human complement proteins such as C3b and iC3b were degraded by Acanthamoeba M28AP. We believe that M28AP is an important factor in human innate immunity. This study provides new insight for the development of more efficient medicines to treat Acanthamoeba infection.

Author Correction: Major ocular trauma in Taiwan: 2002-2004 versus 2012-2014.

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Major ocular trauma in Taiwan: 2002-2004 versus 2012-2014.

We investigated the temporal changes in major eye injuries in Taiwan by reviewing the medical records of all patients with ocular trauma hospitalized at the National Cheng Kung University Hospital during 2002-2004 and 2012-2014. A total of 169 eyes (161 patients) during 2002-2004 and 121 eyes (120 patients) during 2012-2014 were enrolled (mean ± SD age: 41.9 ± 20.8 years in 2002-2004, and 51.8 ± 19.3 years in 2012-2014). Males accounted for ~75% of patients. The most frequent injury-causing object was metallic material (~24%), and blunt traumas were most frequently attributable to traffic accidents and falls. The most common locations of injuries for males and females were the workplace and home, respectively. Open-globe injuries occurred in ~70% of eyes, requiring primary repair, cataract extraction, and/or intraocular lens implantation. The frequencies of fall injury, lacrimal system laceration, lens injury, corneal/scleral foreign bodies, and use of intracameral antibiotics increased from 2002-2004 to 2012-2014, while those of closed-globe injury, vitreous haemorrhage, optic nerve injury, and medical treatment decreased. The final visual acuity remained poor (≤20/200) in >1/3 of injured eyes. Despite therapeutic advancements, major eye injuries still pose a high risk for poor visual outcome.

Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan.

BACKGROUND/PURPOSE: Acanthamoeba keratitis (AK), a painful infectious corneal disease, is caused by the free-living pathogenic species Acanthamoeba. The symptoms include corneal infiltrate, epithelial, and stromal destruction, and loss of vision. Current treatment generally involves an hourly application of polyhexamethylene biguanide (PHMB) over a period of several days; however, even this is not entirely effective against all strains/isolates. The aims of this study were to confirm the existence of pathogenic strains in Taiwan which are highly resistant to drugs and to characterize the behavior of these strains. METHODS: An in vitro Acanthamoeba species culture platform was established to observe the effectiveness of treatment and chart the morphological changes that occur under the effects of drugs using a light microscope and time-lapse recording. Changes in gene expression were examined using reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. RESULTS: Over 90% of the standard strain cells (ATCC 30010) were lysed after being treated with PHMB for 1 hour; however, clinical isolates of Acanthamoeba castellanii that differed in their susceptibility to the treatment drug were only partly lysed. Following treatment with PHMB, National Cheng Kung University Hospital isolation B (NCKH_B) transformed into a pseudocyst under the effects of drug stress; however, National Cheng Kung University Hospital isolation D (NCKH_D), an isolate with higher tolerance for PHMB, did not transform. CONCLUSION: Our results confirm the existence of clinical isolates of A. castellanii with high resistance to PHMB in Taiwan and present the alternative drug tolerance of A. castellanii in addition to the transformation of pseudocyst/cyst.

The effect of the disulfideisomerase domain containing protein in the defense against polyhexamethylene biguanide of highly tolerant Acanthamoeba at the trophozoite stage.

Acanthamoeba castellanii is a free-living protozoan pathogen capable of causing a blinding keratitis and fatal granulomatous encephalitis. Current treatment generally involves an hourly application of polyhexamethylene biguanide (PHMB) over a period of several days but this is not entirely effective against all strains/isolates. The tolerance mechanisms of PHMB in Acanthamoeba cells remain unclear. In this study, we found that the mRNA expression level of disulfideisomerase domain containing protein (PDI) increased rapidly in surviving cells of the highly PHMB-tolerant Acanthamoeba castellanii strain, NCKH_D, during PHMB treatment, but not in the ATCC standard strain. After PDI-specific silencing, NCKH_D was found to be more vulnerable to PHMB treatment. The results described above show that PDI is an important gene for PHMB tolerance ability in a highly PHMB-tolerant strain of Acanthamoeba and provide a new insight for more efficient medicine development for Acanthamoeba keratitis.

Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain.

Acanthamoeba keratitis (AK) is a serious ocular disease caused by pathogenic Acanthamoeba gaining entry through wounds in the corneal injury; generally, patients at risk for contracting AK wear contact lenses, usually over a long period of time. Moreover, pathogenic Acanthamoeba causes serious consequences: it makes the cornea turbid and difficult to operate on, including procedures such as enucleation of the eyeball. At present, diagnosis of this disease is not straightforward, and treatment is very demanding. We have established the comparative transcriptome and extracellular secreted proteomic database according to the non-pathogenic strain ATCC 30010 and the pathogenic strains NCKU_B and NCKU_D. We identified 44 secreted proteins successfully, 10 consensus secreted proteins and 34 strain-specific secreted proteins. These proteins may provide targets for therapy and immuno-diagnosis of Acanthamoeba infections. This study shows a suitable approach to identify secreted proteins in Acanthamoeba and provides new perspectives for the study of molecules potentially involved in the AK.

A cluster of endophthalmitis caused by Mycobacterium abscessus after cataract surgery.

We report two cases of postoperative endophthalmitis after cataract surgery caused by the same strain of Mycobacterium abscessus confirmed by arbitrarily primed polymerase chain reaction, sequencing of the erythromycin ribosome methyltransferase gene and pulsed-field gel electrophoresis. The outcomes were poor despite aggressive treatments. This is the first report of nontuberculous mycobacteria as a causative pathogen for a cluster of endophthalmitis.

Spontaneous suprachoroidal hemorrhage: Case report and review of the literature.

We present the clinical course, management, and final outcome of spontaneous suprachoroidal hemorrhage (SSCH) in an age-related macular degeneration (AMD) patient-a 64-year-old male receiving antiplatelet therapy who developed SSCH during the Valsalva maneuver. In addition to our case study, we discuss the results of a systemic review of the literature and reference lists of retrieved studies published from January 2001 to December 2013. Among a total of 31 patients (32 eyes), acute secondary glaucoma was a complication in 87.5% of the cases, and over half of the cases (20 eyes, 62.5%) received surgery. Twenty cases (64.5%) were characterized by systemic hypertension (HTN), followed by cardiovascular or cerebral vascular disease in 17 cases (54.8%). The Valsalva maneuver was performed in five cases (16.1%) prior to the episode. Twenty-three cases (74.2%) had abnormal hemostasis, including use of anticoagulants or thrombolytic agents (18 cases), chronic renal failure (CRF, 5 cases), and blood dyscrasia (3 cases). AMD was the most common (17 eyes of 16 patients, 53.1%) ocular disease. Visual acuity was classified as hand motion (HM) or worse in 20 eyes (of 28 eyes, 71.4%) at initial presentation and in 24 eyes (of 30 eyes, 80%) upon final examination. Anticoagulated patients with AMD should be informed of the risk of intraocular hemorrhage. Medical therapy usually fails in the treatment of glaucoma. Surgical intervention provides an option for the purpose of pain relief. Even so, the final visual prognosis is usually poor.

Refractive Changes in Amblyopic Children with High Anisometropia.

PURPOSE: To demonstrate the refractive changes of amblyopic children attributed to high anisometropia in the myopia endemic in Taiwan. METHODS: Amblyopic children younger than 10 years with myopic or hyperopic anisometropia 3 diopters (D) or more who had follow-up for more than 2 years and had final visual acuity of 20/30 or better in the amblyopic eye were included. RESULTS: The average age on the first visit was similar for the 13 myopic children (5.5 years) and 17 hyperopic children (5.3 years). Initially, the mean anisometropia was significantly different (myopic, 7.25 D vs. hyperopic, 3.89 D), but the visual acuity of the amblyopic eye (logarithm of the minimum angle of resolution) was similar between both groups at baseline (0.50 vs. 0.57). Myopization of the sound eye surpassed that of the amblyopic eye in the myopic group, which resulted in a yearly reduction of anisometropia of 0.51 D. However, hyperopia decreased synchronously in both eyes of the hyperopic children. The final degree of anisometropia was not significantly different between the two groups (4.07 D vs. 3.62 D). CONCLUSIONS: Growing up in a country with a high prevalence of myopia, children with myopic and hyperopic anisometropia in this study inevitably had myopization in both eyes. Myopic anisometropia decreases significantly over time, but hyperopic anisometropia remains constant.

Repetitive methylene blue-mediated photoantimicrobial chemotherapy changes the susceptibility and expression of the outer membrane proteins of Pseudomonas aeruginosa.

BACKGROUND: Since bacterial multidrug efflux pumps mediate intracellular photosensitizer methylene blue, a change in the expression alters the susceptibility to photoantimicrobial chemotherapy (PACT) of Pseudomonas aeruginosa, which may occur following repetitive sublethal challenges. MATERIALS AND METHODS: We performed 10 consecutive, methylene blue-mediated PACT on one antibiotic-sensitive strain and three antibiotic-resistant strains of P. aeruginosa. Following each therapy, the surviving bacteria were collected for subsequent PACT. The susceptibility was compared for the pre- and the post-treated strains following repetitive PACT. To explore the existence of efflux pumps, one of the inhibitors, namely Phe-Arg ß-naphthylamide dihydrochloride (PAßN 25 µg/ml), was added. Profiles of outer membrane proteins were obtained for the pre-treated and the post-treated strains. RESULTS: The susceptibility of PACT did not correlate with the antibiotic sensitivity. Following ten PACT, there was no significant change in susceptibility for three tested strains, except for one antibiotic-resistant strain, for which the 10th generation became less susceptible than the original one. With 2-D electrophoresis, a change in the expression of outer membrane proteins was observed. PAßN successfully increased the phototoxicity in all tested strains, especially the less PACT-susceptible 10th generation of the antibiotic-resistant strain. CONCLUSIONS: Following repetitive challenges, PACT had a consistent antimicrobial effect on three strains; however, one antibiotic-resistant strain, which was the most vulnerable to PACT, became more resistant after consecutive challenges. In addition, the post-PACT strain had different expression of outer membrane proteins, providing further evidence view that repetitive PACT with methylene blue could change the expression of efflux pumps.

Retinopathy of prematurity in southern Taiwan: a 10-year tertiary medical center study.

BACKGROUND/PURPOSE: Retinopathy of prematurity (ROP) is a leading cause of childhood blindness. This retrospective study investigated ROP, including incidence, demographic information,risk factors, treatments, and refractive outcomes, in southern Taiwan over a 10-year period. METHODS: The authors retrieved the National Cheng Kung University Hospital database between the years 2000 and 2009 for newborns with a gestational age less than 32 weeks and/or with a birth weight less than 1500 g who had been screened for ROP. We recorded sex, birth weight, gestational age, in-hospital versus out-of-hospital birth, paternal and maternal ages, whether there were multiple gestations, parity, Apgar scores, length of hospital stay, risk factors, presence and severity of ROP and whether it was treated, and refraction at the last visit. Regression analyses were performed to identify risk factors for ROP. RESULTS: A total of 503 live births were included. ROP was identified in 190 (37.8%) and met criteria for treatment in 59 (11.7%).ROP was diagnosed as stage 1, 2, 3, 4, and 5 in 61 (12.1%), 36 (7.2%), 81 (16.1%), 11 (2.2%), and 1 (0.2%) infant, respectively. Lower birth weight and younger gestational age were risk factors for greater severity of ROP (p < 0.001). Of the 167 with extremely low birth weight (<1000 g), 118 (70.7%) had ROP and 49 (29.3%) required treatment. On univariate analysis, low birth weight, younger gestational age, and risk factors such as respiratory distress syndrome, chronic lung disease, patent ductus arteriosus, surfactant usage, indomethacin usage, sepsis, upper gastrointestinal bleeding, blood transfusion, and necrotizing enterocolitis were associated with ROP. Multivariate logistic regression analysis showed that only lower birth weight was a significant and independent risk factor for ROP. Myopia (76%)and anisometropia (28%)were common in advanced ROP. CONCLUSION: Low birth weight is a major risk factor for ROP. Infants with extremely low birth weight had a higher risk of severe ROP. Common ocular sequelae of advanced ROP were myopia and anisometropia.

A preequatorial Gore-Tex sling to restore rectus muscle pathways in myopic strabismus fixus.

A 74-year-old woman with high myopia presented with a rapid-onset, large-angle esotropia consistent with myopic strabismus fixus, or fallen eye syndrome. We treated the patient with combined bilateral disinsertion of the medial rectus muscles and resection of the right lateral rectus muscle with bilateral repositioning of the displaced superior and lateral rectus muscles toward their normal anatomic positions using a polytetrafluoroethylene (Gore-Tex) sling. The eyes were held in the orthotropic position for 7 days after surgery using nylon traction sutures. After 7 months of follow-up, the patient remained orthotropic at distance with a residual esotropia of 12(Δ) at near and a persistent limitation of elevation.

Treatment of early Acanthamoeba keratitis with alcohol-assisted epithelial debridement.

PURPOSE: To study the safety and efficacy of treating early-stage Acanthamoeba keratitis (AK) with 20% alcohol-assisted epithelial debridement. METHODS: Four consecutive patients (2 wearing orthokeratology lenses and 2 wearing soft contact lenses) presented with pseudodendrites, radial keratoneuritis, and epithelial irregularities. Using a technique similar to laser-assisted subepithelial keratomileusis, we performed alcohol-assisted full-thickness debridement of the corneal epithelium and sent portions for smears, histopathologic and ultrastructural examinations, and culture for evidence of Acanthamoeba. Patients were then started on topical propamidine isethionate and 0.02% polyhexamethylene biguanide. RESULTS: Immediately after debridement, minimal underlying anterior stromal infiltrate or haze was seen. Dosages of antiamoebic agents were tapered as corneal defects reepithelialized (in 1-3 weeks) with no evidence of post-debridement corneal infection. At the final follow-up, 1 cornea was transparent and the other 3 corneas had very faint subepithelial haze. Cultures of all epithelial debridement specimens yielded Acanthamoeba trophozoites and cysts, and histopathologic and electron microscopic examinations revealed Acanthamoeba organisms within corneal epithelial layers. CONCLUSIONS: Alcohol-assisted epithelial debridement facilitates detachment of the full-thickness corneal epithelial layer in a controlled manner and seems to be effective in the treatment of early-stage AK. Unlike the fragile fragmented specimens obtained by mechanical scraping without alcohol soaking, epithelial sheets detached easily and the architectures were well preserved, permitting histopathologic and ultrastructural examinations. Most importantly, 20% alcohol-assisted epithelial debridement did not prevent culturing of Acanthamoeba from the removed epithelial specimens.

Effects of photodynamic therapy on rapidly growing nontuberculous mycobacteria keratitis.

PURPOSE: The authors investigated the antimicrobial effect of methylene blue (MB)-mediated photodynamic therapy (PDT) on Mycobacterium fortuitum keratitis. METHODS: In the in vitro study, the mycobacterial suspension and colonies were treated with the following: no MB, no light (normal control); MB and no light (dye control); light and no MB (light control); MB and light (PDT). Morphologic characteristics were examined by transmission electron microscopy. The bactericidal effects of combined PDT and antibiotic therapy (ciprofloxacin, moxifloxacin, and amikacin) were determined using the broth microdilution technique. Twenty-one rabbits with Mycobacterium keratitis were randomly divided into three groups (no treatment, topical amikacin treatment, and PDT combined with amikacin treatment). The clinical features of keratitis were scored and graded before treatment and before euthanatization. The diseased corneas were trephined for quantitative bacteriologic analysis to determine the antibacterial efficacy of the treatment. RESULTS: In the in vitro tests, the bacterial count had a 2-log reduction immediately after PDT treatment at 100 J/cm(2) with 10(-3)% MB. After PDT at 100 J/cm(2) with 10(-2)% MB, almost no viable bacteria were detected. PDT had a synergistic antimicrobial effect in combination with antibiotics. The phototoxicity occurred in the cytoplasm first and then disrupted the mycobacterial cell walls by lysis. In the rabbit keratitis model, combined PDT resulted in significantly less bacterial burden (P < 0.01) than in the amikacin group. CONCLUSIONS: This study demonstrated the effectiveness of MB-mediated PDT against Mycobacterium fortuitum. PDT could be a potential alternative treatment for nontuberculous mycobacterial corneal infections.

Paederus-induced keratitis.

PURPOSE: To report the clinical picture and management of 4 cases of presumed paederus-induced keratitis. METHODS: The clinical characteristics of the anterior segment and periocular area were presented. The outcomes and healing course of the corneal lesions were recorded. RESULTS: Each patient sustained an accidental eye injury from a beetle while riding a motorcycle. Three cases were complicated by periocular dermatitis. All patients had diffuse punctate epithelial erosions or large epithelial defects on the unilateral cornea. Prolonged corneal epithelialization was noted after lubrication with artificial tears and wearing a contact lens. Reepithelialization was promoted after topical supplementation with 20% autologous serum. All 4 patients had good visual outcomes. Two patients healed with a faint corneal scar. CONCLUSIONS: Paederus-induced keratitis could result in extensive involvement of the corneal surface. Keratitis with a large epithelial defect mimicking the corrosive lesion of a chemical burn requires aggressive therapy, such as autologous serum combined with a bandage contact lens. Avoiding contact with the crushed body of the beetle while rubbing the eye is mandatory to prevent this type of keratitis.

Major pediatric ocular trauma in Taiwan.

PURPOSE: To investigate major pediatric ocular trauma in Taiwan. METHODS: Retrospective review of medical records of all patients 15 years and younger who were hospitalized with a primary diagnosis of eye injury at National Cheng Kung University Hospital, Taiwan, between June 1988 and May 2006. RESULTS: There were 156 children (156 eyes) 1.1 to 15.0 years (mean+/-standard deviation, 7.1+/-0.3 years; boy: girl ratio: 2.1:1). Objects most often causing penetrating injury were scissors (13.5%), pencils/pens (12.2%), broken eyeglasses/spectacles (7.7%), and knives (6.4%). Most blunt trauma occurred in traffic accidents (5.8%). Most injuries occurred at home, followed by on the street, at school, and at sports venues. Injuries were classified as open globe (71.2%), adnexal only (18.6%), or closed globe (10.3%), and included corneal laceration (40.4%), lens damage (27.6%), hyphema (25.6%), and eyelid laceration (23.7%). Most surgical procedures were primary repair (88.5%) or removal of a damaged lens (22.4%). Additional surgery was performed in 19.9% of cases. After treatment, 56.4% of eyes had corneal opacity/scar and 7.1% became phthitic; 52.6% had good visual outcome, whereas 23.1% had poor final vision. Compared with visual acuity measured on admission, final visual acuity was improved in 76.1%, unchanged in 19.7%, and worse in 4.3%. Predictors of worse outcome were open-globe injury and larger wound size, posterior segment involvement, and presence of an intraocular foreign body. CONCLUSIONS: Most of the children hospitalized for major ocular trauma are younger boys with penetrating injuries suffered at home. Most injuries could have been prevented by increased awareness and reduction of risk factors, and the authors urge better public education for improved safety.

Astigmatism in children with epiblepharon.

PURPOSE: To study the relationship between astigmatism and epiblepharon in Taiwanese children that need surgical correction. METHODS: In a retrospective case-control study, a total of 254 eyes in 134 children surgically treated for lower-lid epiblepharon were studied. A further 205 eyes in 104 children 4-7 years of age were included as controls. Data on age, sex, severity of preoperative corneal erosion, best-corrected visual acuity, and preoperative and postoperative refractive errors were recorded. The data of astigmatism were decomposed into P90 (at 90 degrees) and its oblique meridian Pobl. Nonparametric tests were used to compare the severity of astigmatism. RESULTS: No significant difference between age group (<4, 4-7, >7 years) in P90 was shown in children with epiblepharon. Pobl was trivial in each group of epiblepharon. Children with epiblepharon 4-7 years of age had significantly (P < 0.001) greater P90 than controls (1.12 vs. 0.47 D). P90 and Pobl were not significantly changed postoperatively. In the same individual, the eye with the more severely affected cornea had greater astigmatism (P = 0.002), even after the corneal erosion healed. CONCLUSIONS: Children with epiblepharon who needed surgery had greater with-the-rule astigmatism than controls. More severe corneal erosion was associated with higher astigmatism. Surgical correction of skinfold and promotion of reepithelialization did not attenuate astigmatism significantly.