ß-1,4-Galactosyltransferase III suppresses extravillous trophoblast invasion through modifying ß1-integrin glycosylation.

INTRODUCTION: Glycosylation controls diverse protein functions and regulates various cellular phenotypes. Trophoblast invasion is essential for normal placental development. However, the role of glycosylation in human placenta throughout pregnancy is still unclear. The ß-1,4-galactosyltransferase III (B4GALT3) has been found to regulate cancer cell invasion. We therefore investigated the expression of B4GALT3 in placenta and its roles in trophoblast. METHODS: B4GALT3 protein expression was examined by quantitative Western blotting analysis in human placentas. For identification of B4GALT3-positive cells in normal human placenta, immunohistochemistry and immunofluorescence methods were used. To investigate effects of B4GALT3 on extravillous trophoblast (EVT)-like cell and primary EVT cells, we analyzed cell growth, adhesion, migration, and invasion in mock and B4GALT3-transfected cell. RESULTS: B4GALT3 expression significantly increased in third trimester human placenta. Immunostaining revealed that B4GALT3 expressed in placental villous cytotrophoblast, syncytiotrophoblast, and a subpopulation of EVT cells throughout pregnancy. Interestingly, we found increases in the expression level and percentage of B4GALT3-positive cells in third trimester EVT, but not in syncytiotrophoblasts and cytotrophoblasts of placental villi. Overexpression of B4GALT3 in HTR8/SVneo cells and primary trophoblast cells significantly suppressed cell migration. In addition, B4GALT3 suppressed cell invasion, and enhanced cell adhesion to laminin in HTR8/SVneo cells. Notably, we found that B4GALT3 modified glycans on ß1-integrin, suppressed focal adhesion kinase (FAK) signaling, and enhanced ß1-integrin degradation. DISCUSSION: We propose that B4GALT3-mediated glycosylation change not only enhances ß1-integrin binding to laminin, but also attenuates ß1-integrin stability. Our findings suggest that B4GALT3 is a critical regulator for suppressing EVT invasion in the late stages of pregnancy.

Astigmatism and biometric optic components of diode laser-treated threshold retinopathy of prematurity at 9 years of age.

PURPOSE: To assess the prevalence of astigmatism and its relationship with biometric optic components in preterm school children with diode laser-treated threshold retinopathy of prematurity (ROP). METHODS: A prospective, cross-sectional study in which cycloplegic keratometry, refraction, and ultrasound biometric measurement of optic components were performed on 24 consecutive preterm children with diode laser-treated threshold ROP at the age of 9 years. The study results were compared with data on 1021 age-matched full-term control children from a national survey. RESULTS: The laser-treated eyes had a mean astigmatism of 3.47 D, with a mean spherical equivalent of -4.49 D. Of the 46 eyes studied, 98% of eyes showed astigmatism ≥0.5 D and 50% had high astigmatism (>3.0 D). Most astigmatic eyes (97.7%) showed with-the-rule astigmatism, with the mean plus cylinder axis at 89.30(o). Further correlation analysis showed the astigmatism in refraction was highly correlated with the corneal astigmatism (r=0.921, P<0.001) and the vertical corneal curvature (r=0.405, P=0.005). There was significantly steeper vertical corneal curvature (P=0.003) and flatter horizontal corneal curvature (P=0.031) in eyes with laser-treated ROP when compared with age-matched full-term controls. The eyes with laser-treated ROP also show significantly thicker lens (3.93 mm) and shallower anterior chamber depth (ACD; 2.92 mm) than full-term controls (P<0.001). CONCLUSIONS: There is significantly higher prevalence and greater magnitude of astigmatism in eyes with laser-treated threshold ROP compared with full-term controls. The steeper vertical corneal curvature component contributes to the increased astigmatism in eyes with laser-treated ROP.

Increased oxidative DNA damage, lipid peroxidation, and reactive oxygen species in cultured orbital fibroblasts from patients with Graves' ophthalmopathy: evidence that oxidative stress has a role in this disorder.

PURPOSE: We investigated the oxidative stress in orbital fibroadipose tissues and cultured orbital fibroblasts from patients with Graves' ophthalmopathy (GO). METHODS: The content of 8-hydroxy 2'-deoxyguanosine (8-OHdG), an important biomarker of oxidative DNA damage, was measured in orbital fibroadipose tissues and cultured orbital fibroblasts from patients with GO and compared with age-matched normal controls. A product of lipid peroxidation, malondialdehyde (MDA), and intracellular reactive oxygen species (ROS) in cultured orbital fibroblasts was also determined. RESULTS: There was no significant difference in the 8-OHdG content of orbital fibroadipose tissues between patients with GO and age-matched normal controls (P=0.074). However, the levels of 8-OHdG and MDA in GO orbital fibroblasts were significantly higher than those of normal controls (P=0.0026 and P<0.001, respectively). In addition, GO orbital fibroblasts had higher contents of superoxide anions and hydrogen peroxide compared with those of normal controls (P=0.0133 and 0.0025, respectively). CONCLUSIONS: Orbital fibroblasts represent the most abundant cell type among orbital connective tissues and exhibit great differences in their phenotypes. Increased oxidative DNA damage and lipid peroxidation, as well as higher intracellular ROS levels in GO orbital fibroblasts may have a role in the pathogenesis of GO.

Use of retrograde esophagoscopy in delayed primary esophageal anastomosis for isolated esophageal atresia.

INTRODUCTION: Preserving the native esophagus is critical for long-term swallowing function in patients with esophageal atresia (EA). However, long esophageal gaps and hidden distal esophageal pouches are frequently encountered, making primary esophageal anastomosis very difficult in cases with isolated EA. This study evaluates the efficacy of retrograde esophagoscopy for the identification of distal esophageal pouches to aid primary esophageal anastomosis in patients with isolated EA. MATERIAL AND METHODS: From January 1995 to January 2007, five patients with isolated EA out of 30 patients with EA treated in our hospital were included in this study. All patients initially received a gastrostomy and distal esophagogram to evaluate distal esophageal pouches and esophageal gaps. Delayed esophageal reconstruction was performed 3 to 4 months later. During surgery for esophageal reconstruction, a 0.5 cm diameter endoscope was inserted through the gastrostomy to identify the distal esophageal pouch. RESULTS: Distal esophagograms found no distal esophageal pouch in 3 patients. Retrograde esophagoscopy and exploratory surgery found no distal esophageal pouch in only 1 patient. The esophageal gap ranged from 4 to 7 cm. All patients successfully received primary esophageal anastomosis except for one without a distal pouch who received colon interposition. Postoperative complications included esophageal stricture in 4 patients and gastroesophageal reflux (GER) in 3. All esophageal strictures resolved after esophageal dilatation. One patient required further fundoplication for GER. CONCLUSIONS: Retrograde esophagoscopy is superior to distal esophagogram for the identification of distal esophageal pouches in isolated EA. In addition, retrograde esophagoscopy is excellent for the localization of distal esophageal pouches to facilitate primary end-to-end esophageal anastomosis.

Long-term visual outcomes of laser-treated threshold retinopathy of prematurity: a study of refractive status at 7 years.

PURPOSE: To assess the long-term visual outcomes and refractive status in patients with diode laser-treated threshold retinopathy of prematurity (ROP), and to investigate the causes of impaired visual function. METHOD: A total of 60 eyes of 30 consecutive patients with diode laser-treated threshold ROP were recalled for assessment at the age of 7 years or more. RESULTS: There were 38 eyes (65.5%) achieving 6/12 or better vision, however, an unfavourable visual outcome (6/60 or worse) occurred in four eyes (6.9%). One eye (1.7%) had unfavourable structural outcome. Of these 60 laser-treated eyes, 46 eyes (77.0%) were myopic, the overall mean spherical equivalent was -3.87 D. Anisometropia (>or=1.5 D) was also noted in 14 patients (46.7%). Strabismus was present in nine patients (30.0%). Perinatal neurological events of intraventricular haemorrhage (IVH) were identified in eight children (26.7%), periventricular leucomalacia (PVL) in eight children (26.7%), and cerebral palsy (CP) in four children (13.3%). There was a statistically significant association of the presence of strabismus with PVL (P=0.002). The presence of anisometropia was a significant risk factor associated with poor visual outcome of 6/15 or worse in laser-treated ROP (P=0.002). CONCLUSION: The majority of patients with diode laser-treated threshold ROP had favourable anatomical and visual outcomes. However, anisometropia, advanced refractive error, strabismus, and perinatal neurological events remain important causes of impaired visual function. Long-term follow-up is very important for early detection and timely treatment of these ocular morbidities.

Retinoblastoma in Taiwan: incidence and survival characteristics from 1979 to 2003.

PURPOSE: To describe the incidence and survival rate of patients with retinoblastoma in Taiwan over a 25-year period, from 1979 to 2003. METHODS: Cases of retinoblastoma have been reported to the Taiwan National Cancer Registry (TNCR) since 1979. Retinoblastoma data from 1979 to 2003 were retrieved from the TNCR databank for analysis of both the incidence and survival rate. Trends in the incidence rate were estimated by calculating the annual percentage change (APC). RESULTS: From 1979 to 2003, a total of 380 patients (182 [47.9%] females) with retinoblastoma were identified. Of the 380 cases, 359 patients (94.5%) were diagnosed before 5 years of age. The average annual age-standardized incidence rate was 4.45 per million in children under 10 years of age and 8.58 per million in children under 5 years of age. There was a significant increasing trend in the incidence rate over the study period with an APC of 2.3% (P=0.035). Before the implementation of Taiwan's National Health Insurance (NHI) Program in 1995, the mean age at diagnosis was 4.8+/-13.3 years and the 5-year survival rate was 74.7%. After implementation of the NIH Program, the mean age at diagnosis decreased to 1.6+/-2.1 years (P<0.001) and the 5-year survival rate increased to 84.7% (P=0.063). CONCLUSIONS: We report the first population-based study on retinoblastoma in Taiwan. There was a significant increasing trend in the incidence rate from 1979 to 2003. Since the implementation of Taiwan's NHI, the mean diagnostic age decreased.

Oxidative stress in patients with Graves' ophthalmopathy: relationship between oxidative DNA damage and clinical evolution.

PURPOSE: To evaluate the relationship between oxidative stress and clinical evolution in patients with Graves' ophthalmopathy (GO). METHODS: Thirty-one euthyroid GO patients and 25 healthy subjects participated in this study. Oxidative DNA damage was assessed by determination of the 8-hydroxy-2'-deoxyguanosine (8-OHdG) level in urine by ELISA. The relationship of oxidative DNA damage to the clinical evolutions of GO, especially the smoking status, clinical activity scores (CAS), and ophthalmopathy index was examined. RESULTS: The mean 8-OHdG was significantly higher in GO patients than that of normal controls (12.6+/-5.7 vs 6.7+/-2.5 ng/mg creatinine, P<0.001). Smokers had significant higher 8-OHdG than did never smokers in GO patients (P=0.029), but not in healthy controls (P=0.374). Among GO patients, only CAS remained significantly correlated with 8-OHdG (P=0.001) after adjusting for age, sex, disease duration, the status of antithyroid drug and smoking, and thyroid-stimulating hormone level. Patients with active GO (CAS>3) had higher 8-OHdG than did the patients with CAS

Early and late endophthalmitis following trabeculectomy in a Chinese population.

PURPOSE: To investigate the rate of bleb-related endophthalmitis over 5 years in a Chinese population. METHODS.:Retrospective chart review. RESULTS: Of 988 trabeculectomies performed over 5 years, one case (0.1%) developed early endophthalmitis caused by Morganella morganii, which was rarely reported in the literature. Six cases (0.6%) developed late-onset endophthalmitis. Mitomycin C significantly increased the risk of late-onset endophthalmitis (p=0.0002). CONCLUSIONS: Physicians should weigh the benefits against the risks of mitomycin C application in performing trabeculectomies.

Cytotoxicity of silicone oil on cultivated human corneal endothelium.

BACKGROUND AND OBJECTIVE: To investigate the cytotoxic effects of silicone oil on the cultivated human corneal endothelial cells (CEs). METHODS: We cultured human CE and passed them in insert wells that allowed the apical side of CE monolayer in contact with the silicone oil. The tested silicone oils were of two different viscosities, 1,000 and 5,000 centistoke (CS). MTS proliferation bioassay and calcein-acetoxymethyl ester (CAM)-ethidium homodimer staining were performed to evaluate cell viability after CEs were co-cultured with silicone oils for 48 h. Apoptosis of CEs was evaluated by TdT-mediated dUTP nick-end labelling (TUNEL) stain. RESULTS: The MTS bioassay showed that contact of silicone oil inhibited CE proliferation. The higher viscosity (5,000 CS) silicone oil suppressed cell cycling significantly more than the lower viscosity (1,000 CS) silicone oil. CAM-ethidium homodimer staining revealed CE death, 9.1+/-0.1% (1,000 CS silicone oil) and 41.6+/-0.4% (5,000 CS), but apoptosis played only minor role in silicone oil toxicity, 1.7+/-0.1% (1,000 CS silicone oil) and 9.4+/-0.1% (5,000 CS). CONCLUSIONS: Silicone oil is cytotoxic to cultivated human CEs. Avoiding the forward migration of silicone oil to the anterior chamber and corneal CE contact is critical in preventing silicone oil-associated keratopathy. Silicone oil should be removed as early as possible once the goal of tamponade has been achieved.

Exophthalmos of patients with Graves' disease in Chinese of Taiwan.

PURPOSE: To determine the exophthalmos measurements in patients with Graves' disease (GD) and to compare with those of normal subjects among the Chinese of Taiwan. METHODS: Hertel's exophthalmometry were performed in 163 consecutive adult GD patients and 419 normal subjects. The values of exophthalmos, asymmetry, and the distance between lateral orbital rims (base) were evaluated. RESULTS: There was a significant difference between GD and normal adults for exophthalmos and asymmetry, with GD patients demonstrating a mean exophthalmos of 18.32 mm and asymmetry of 0.93 mm as compared with 13.91 and 0.55 mm for normal subjects (P<0.001). The mean value of base for GD adults (106.52 mm) is slightly greater than normal subjects (105.82 mm). There was no gender difference in exophthalmometric value or asymmetry in each group. A total of 79 (48.5%) GD patients had exophthalmos greater than the upper limit of normal Chinese in Taiwan (18.6 mm). Asymmetry over 2 mm was observed in 11 (6.7%) patients with GD but in none of the normal subjects. CONCLUSIONS: GD patients have significant exophthalmos and asymmetry compared to normal subjects. In comparison with previous studies, exophthalmos values for Chinese in Taiwan tend to be lower than those of Caucasians and black-American people, but are close to those of Korean, Japanese, Iranian, Indian, and Asian-American people. Adequate adjustment of normal and abnormal values of exophthalmos according to individual differences is important for the diagnosis of orbital morbidity and management of thyroid associated orbitopathy.

Increased oxidative DNA damage, 8-hydroxydeoxy- guanosine, in human pterygium.

PURPOSE: Chronic exposure to ultraviolet (UV) light is a widely accepted aetiological factor in the development of pterygium. UV radiation may induce production of reactive oxygen species via photosensitized oxidation, thus causing oxidative damage. This study was conducted to test the hypothesis that oxidative damage to DNA is increased in pterygium. METHODS: Immunohistochemical analysis employing a monoclonal antibody specific for 8-hydroxy-2'-deoxyguanosine (8-OHdG), a ubiquitous maker of oxidative stress, was performed in three patients with primary pterygium. The levels of 8-OHdG in DNA isolated from the other 29 pterygium specimens and their adjacent normal conjunctival tissues were determined using enzyme-linked immunosorbent assay (ELISA). RESULTS: Immunohistochemistry of 8-OHdG showed a distinct pattern of more extensive and intense staining in the nuclei of pterygium tissue compared with that in their adjacent normal conjunctiva. ELISA also revealed that the average level of 8-OHdG in the pterygium tissues was 4.7-fold higher than that of the corresponding normal conjunctiva (P<0.001). CONCLUSIONS: The increased levels of 8-OHdG in the pterygium tissues indicate that oxidative stress could play a role in the development of pterygium. These findings provide new information to better understand the pathogenesis of pterygium and are useful in the prevention and treatment of this disease.

Pulsatile ocular blood flow change after treatment with systemic steroid in patients with Graves' ophthalmopathy.

PURPOSE: Our previous study has demonstrated an impairment of pulsatile ocular blood flow (POBF) in patients with Graves'ophthalmopathy (GO). Here we further evaluate the haemodynamic change in GO patients after systemic steroid. METHODS: In a prospective, interventional, consecutive clinical case series, all patients with active and moderately severe GO who underwent systemic steroid treatment were evaluated. The change of POBF and the clinical activity and severity of the disease were assessed. RESULTS: In total, 11 patients underwent intravenous methylprednisolone pulse therapy followed by 2-month oral prednisolone therapy. POBF improved from a mean value of 476.5 to 614.7 microl/min (P<0.001) after treatment. Likewise, the mean clinical activity score reduced from 4.8 to 1.6. However, systemic steroid had less effect on the severity of GO. It also revealed that there is a greater improvement of POBF in those with more clinical activity score (>4) at the onset. CONCLUSIONS: Following treatment with systemic steroid we have demonstrated a significant improvement in POBF in patients with GO.