ABSTRACT
The study aimed to investigate the extended effects and interrelations of corneal biomechanics, corneal optical density (COD), corneal thickness (CT), and intraocular pressure (IOP) following cataract surgery. Sixteen eyes were analyzed prospectively. The Corneal Visualization Scheimpflug Technology (Corvis ST) device assessed corneal biomechanics, while the Pentacam AxL® (Pentacam) measured COD and CT. Postoperative data were collected around six months after surgery, with a subgroup analysis of data at nine months. The Pearson correlation was used to examine the relationship between surgical-induced changes in corneal biomechanics and COD. At six months, significant postoperative differences were observed in various biomechanical indices, including uncorrected IOP (IOPuct) and biomechanics-corrected IOP (bIOP). However, many indices lost statistical significance by the nine-month mark, suggesting the reversibility of postoperative corneal changes. Postoperative COD increased at the anterior layer of the 2-6 mm annulus and incision site. The changes in COD correlated with certain biomechanical indices, including maximal (Max) deformative amplitude (DA) and stiffness parameter (SP). In conclusion, despite significant immediate postoperative changes, corneal biomechanics, COD, and IOP experienced a gradual recovery process following cataract surgery. Clinicians should maintain vigilance for any unusual changes during the short-term observation period to detect abnormalities early.
ABSTRACT
PURPOSE: To compare corneal haze between active ulcer and healed scarring using a Scheimpflug densitometry. MATERIALS AND METHODS: A prospective longitudinal study enrolled 30 patients (30 eyes) with ulcerative keratitis (UK). Each subject's corneal optical density (COD) was measured with a Scheimpflug corneal densitometry, Pentacam® AXL (Oculus GmbH, Wetzlar, Germany), at the active ulcerative and complete scarring stage. The COD data were analyzed through distinct methods (inbuilt, sorted annular partitions, and ulcer-matching densitometric maps). We compared different CODs to select the better index for clinically monitoring the transition from corneal ulceration to healed scar. RESULTS: The CODs of the periphery (P = 0.0024) and outside of the active ulcer (P = 0.0002) significantly decreased after scarring. Partitioning the cornea into different depths and annular zones, the anterior layer, center layer, and the 2-6 mm annular zone had a more remarkable COD decrease after scar formation. The 3rd-sorted COD in the anterior layer revealed the highest area under the receiver-operating characteristic curves (0.709), in which 90% of subjects had COD reduction during the ulcer-to-scar transition. CONCLUSIONS: Aside from subjective judgment based on clinical signs, the Scheimpflug tomography-based densitometry could provide objective and efficient monitoring of the corneal opacity evolution in UK patients. Because the 3rd-sorted annular COD is a better index than the inbuilt or mapping CODs in differentiating active ulcers from healed scars, this COD could be a clinically promising parameter to monitor the progression of UK patients.
Subject(s)
Cornea , Corneal Ulcer , Densitometry , Humans , Prospective Studies , Female , Male , Densitometry/methods , Middle Aged , Corneal Ulcer/diagnosis , Cornea/pathology , Cornea/diagnostic imaging , Follow-Up Studies , Adult , Wound Healing , Cicatrix/diagnosis , Cicatrix/etiology , Aged , ROC Curve , Visual Acuity , Corneal Topography/methods , Corneal Opacity/diagnosis , Corneal Opacity/etiology , Corneal Opacity/physiopathologyABSTRACT
Previous studies have highlighted the importance of outdoor time in reducing the risk of myopia progression. Although ultraviolet A (UVA) radiation dominates in terms of energy with respect to the UV radiation reaching the Earth's surface, its effects on the exposed anterior sclera have not been well studied. This study was designed to investigate the UVA-induced biological effects at peak sunlight levels in human scleral fibroblasts (HSFs). Using next-generation sequencing (NGS), we analyzed the differentially expressed genes (DEGs) in UVA-treated and normal HSFs. Further, we then identified the functions and key regulators of the DEGs using bioinformatics analysis, and verified the effects of UVA on gene and protein expression in HSFs using real-time PCR, western blotting, and immunofluorescence imaging. The highest level of solar UVA (365 nm) was 3.4 ± 0.18 (mW/cm2). The results from the functional analysis of the DEGs were related to structural changes in the extracellular matrix (ECM) and protein metabolism. Transforming growth factor-ß1 (TGF-ß1) and Smad3 were predicted to be potential upstream regulators, associated with ECM organization. Exposure to a single wavelength of UVA (365 nm, 3 mW/cm2) for 1 h for 5 consecutive days induced the downregulation of the mRNA of ECM genes including COL1A1, COL3A1, COL5A1, VCAN and collagen I protein in HSF. UVA downregulated Smad3 protein and reduced TGF-ß-induced collagen I protein production following UVA exposure in HSF. In conclusion, high UVA exposure reduces TGF-ß signaling and collagen I production by modulating Smad levels in HSF. The effects of overexposure to high-intensity UVA on myopia control require further investigations.
Subject(s)
Myopia , Transforming Growth Factor beta , Humans , Transforming Growth Factor beta/metabolism , Sclera/metabolism , Collagen/metabolism , Transforming Growth Factor beta1/metabolism , Collagen Type I/genetics , Collagen Type I/metabolism , Fibroblasts/metabolism , Ultraviolet Rays/adverse effects , Myopia/metabolism , Transforming Growth Factors/metabolism , Transforming Growth Factors/pharmacologyABSTRACT
To pursue the least corneal implication during cataract surgery, this study aimed to monitor corneal wound injury after cataract surgery with a novel method. The prospective cohort study involved thirty-two patients, who were assessed by a Scheimpflug tomography AxL® (Oculus GmbH, Wetzlar, Germany) via the following two kinds of indices: whole corneal optical density (COD) and corneal thickness (CT), two weeks before and one month after cataract surgery. The results of the COD revealed that corneal annuli 0.0-2.0 mm and 2.0-6.0 mm, and the average and maximal values at the incisional site significantly increased postoperatively. Also, the anterior and central stroma of 0.0-2.0 mm, and all three depths of 2.0-6.0 mm, increased remarkably after the operation. For the CT, all ranges of diameters plus incisional sites showed significant increases postoperatively. Furthermore, we analyzed the differences (delta) of COD and CT between pre- and post-operation, and found significant correlations between the delta of COD and the delta of CT, regarding annuli 0.0-2.0 mm, 2.0-6.0 mm, and 6.0-10.0 mm, but no correlation at the incisional site, with either average density or maximal density, was detected. We concluded that whole COD and CT, especially at the central zones of the cornea (annulus < 6 mm), are both valuable parameters in the assessment of corneal damage post-cataract surgery, and are independent indices at the incisional site.
ABSTRACT
Fibroblast-like transformation of retinal pigment epithelial (RPE) cells is a pathological feature of proliferative vitreoretinopathy (PVR) that may cause blindness. The effect of oxidative stress alone or together with transforming growth factor-beta 2 (TGF-ß2) on epithelial-mesenchymal transformation (EMT) is not fully understood in RPE. TGF-ß2 induced the upregulation EMT markers including α-smooth muscle actin (α-SMA), Snail, and Slug and downregulation of E-cadherin (E-cad) in ARPE-19 cells. Hydrogen peroxide (H2O2) not only upregulated α-SMA but also enhanced the effect of TGF-ß2 on the expression of Snail and Slug. The CXCL family of cytokines could be the mediators of EMT induced by H2O2 and TGF-ß2. H2O2 induced CXCL1, that upregulated α-SMA and fibronectin. Both SB225002, an inhibitor of CXCR2, and antioxidant N-acetylcysteine suppressed the TGF-ß2-induced EMT in ARPE-19 cells. Taken together, the results suggest that oxidative stress enhanced TGF-ß2-induced EMT through the possible autocrine effect of CXCL1 on CXCR2 in ARPE-19 cells.
Subject(s)
Autocrine Communication/drug effects , Chemokine CCL1/biosynthesis , Epithelial-Mesenchymal Transition/drug effects , Oxidative Stress/drug effects , Transforming Growth Factor beta2/pharmacology , Cell Line , Gene Expression Regulation/drug effects , Humans , Hydrogen Peroxide/pharmacology , Phenylurea Compounds/pharmacology , Receptors, Interleukin-8B/antagonists & inhibitors , Receptors, Interleukin-8B/metabolism , Vitreoretinopathy, Proliferative/metabolism , Vitreoretinopathy, Proliferative/pathologyABSTRACT
Graves' orbitopathy (GO) is an autoimmune inflammatory disease affecting the orbit. Orbital fibroblasts are a key component in GO pathogenesis, which includes inflammation, adipogenesis, hyaluronic acid (HA) secretion, and fibrosis. Macrophages are thought to participate in the immunological stage of GO, but whether they can directly affect the fibroblasts phenotype and modulate disease progression is unknown. We previously showed that GO adipogenic and fibrotic phenotypes could be modelled in a pseudo-physiological 3D environment in vitro. Here, we introduced macrophages in this 3D culture model to investigate role for macrophages in modulating adipogenesis, HA production, and contractility in orbital fibroblasts. Macrophages had a minimal effect on lipid droplet formation in fibroblasts, but significantly increased HA production and cell contractility, suggesting that they may promote the fibrotic phenotype. This effect was found to be mediated at least in part through phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) activation and linked to an increase in actin polymerization and protrusive activity in fibroblasts. Overall our work shows for the first time a direct role for macrophages in modulating the fibroblasts' phenotype in GO, supporting a role for macrophages in the progression of the fibrotic phenotype through induction of HA production and stimulation of the contractile phenotype in orbital fibroblasts.
Subject(s)
Cell Communication , Fibroblasts/metabolism , Hyaluronic Acid/biosynthesis , Macrophages/metabolism , Actins/metabolism , Cells, Cultured , Disease Susceptibility , Fibrosis/etiology , Fibrosis/metabolism , Fibrosis/pathology , Graves Ophthalmopathy/etiology , Graves Ophthalmopathy/metabolism , Graves Ophthalmopathy/pathology , Humans , Lipid Metabolism , Macrophages/immunology , Orbit , Phosphatidylinositol 3-Kinases/metabolism , Protein Multimerization , Signal Transduction , Transforming Growth Factor beta/metabolismABSTRACT
PURPOSE: To investigate the effectiveness of a school-based program promoting outdoor activities in Taiwan for myopia prevention and to identify protective light intensities. DESIGN: Multi-area, cluster-randomized intervention controlled trial. PARTICIPANTS: A total 693 grade 1 schoolchildren in 16 schools participated. Two hundred sixty-seven schoolchildren were in the intervention group and 426 were in the control group. METHODS: Initially, 24 schools were randomized into the intervention and control groups, but 5 and 3 schools in the intervention and control groups, respectively, withdrew before enrollment. A school-based Recess Outside Classroom Trial was implemented in the intervention group, in which schoolchildren were encouraged to go outdoors for up to 11 hours weekly. Data collection included eye examinations, cycloplegic refraction, noncontact axial length measurements, light meter recorders, diary logs, and questionnaires. MAIN OUTCOME MEASURES: Change in spherical equivalent and axial length after 1 year and the intensity and duration of outdoor light exposures. RESULTS: The intervention group showed significantly less myopic shift and axial elongation compared with the control group (0.35 diopter [D] vs. 0.47 D; 0.28 vs. 0.33 mm; P = 0.002 and P = 0.003) and a 54% lower risk of rapid myopia progression (odds ratio, 0.46; 95% confidence interval [CI], 0.28-0.77; P = 0.003). The myopic protective effects were significant in both nonmyopic and myopic children compared with controls. Regarding spending outdoor time of at least 11 hours weekly with exposure to 1000 lux or more of light, the intervention group had significantly more participants compared with the control group (49.79% vs. 22.73%; P < 0.001). Schoolchildren with longer outdoor time in school (≥200 minutes) showed significantly less myopic shift (measured by light meters; ≥1000 lux: 0.14 D; 95% CI, 0.02-0.27; P = 0.02; ≥3000 lux: 0.16 D; 95% CI, 0.002-0.32; P = 0.048). CONCLUSIONS: The school-based outdoor promotion program effectively reduced the myopia change in both nonmyopic and myopic children. Outdoor activities with strong sunlight exposure may not be necessary for myopia prevention. Relatively lower outdoor light intensity activity with longer time outdoors, such as in hallways or under trees, also can be considered.
Subject(s)
Leisure Activities , Light , Myopia/prevention & control , Refraction, Ocular/physiology , Schools , Child , Disease Progression , Female , Humans , Incidence , Male , Myopia/epidemiology , Myopia/physiopathology , Odds Ratio , Taiwan/epidemiology , Vision TestsABSTRACT
The purpose of this study was to investigate the spectrum of organisms causing endophthalmitis after cataract surgery in a tertiary medical center in Taiwan and the antibiotic susceptibilities. This was a retrospective case series study. Patients with endophthalmitis after cataract surgery from January 2004 to July 2015 were reviewed. The outcome measures included the identification of isolates, antibiotic susceptibilities, and final visual outcomes. Twenty-one organisms were isolated from 19 cases. The most common organisms were Enterococcus in 38.1 %, especially Enterococcus faecalis, followed by Staphylococcus epidermidis in 28.6 % and Staphylococcus aureus in 9.5 %. All of the Gram-positive isolates tested were susceptible to vancomycin (100 %), and ceftazidime and amikacin were susceptible for Gram-negative organisms. The Gram-positive organisms remain to be the predominant cause of postoperative endophthalmitis, and Enterococcus species has had an increasing incidence. Vancomycin is still the most powerful antibiotic for Gram-positive organisms, while ceftazidime and amikacin are effective for Gram-negative bacteria.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteria/isolation & purification , Cataract Extraction/adverse effects , Eye Infections, Bacterial/drug therapy , Forecasting , Surgical Wound Infection/drug therapy , Vitreous Body/microbiology , Bacteria/drug effects , Endophthalmitis/drug therapy , Endophthalmitis/microbiology , Eye Infections, Bacterial/microbiology , Female , Humans , Male , Microbial Sensitivity Tests , Retrospective Studies , Surgical Wound Infection/microbiology , Visual AcuityABSTRACT
PURPOSE: The toll-like receptor 4 (TLR4) signaling pathway is involved in chronic inflammation and insulin resistance, which are associated with obesity and diabetes mellitus. In the present study, a model of high-fat diet (HFD) feeding of mice was used to investigate the role of TLR4 in overnutrition- and obesity-associated inflammation and infiltration of macrophages and microglia in the retina. METHODS: Wild-type C57BL/6 and TLR4 knockout (TLR4KO; B6.B10ScN-Tlr4(lps-del)/JthJ) mice were fed a HFD or control chow diet (CD) for 6 months. The TLR4 expression, the relative increase in macrophages/microglia (CD11b(+) and CD45(+) cells), the presence of markers of oxidative stress (gp91phox and malondialdehyde; MDA), and DNA damage (phosphorylated histone H2AX; γH2AX) were assessed by real-time PCR and immunofluorescence studies. RESULTS: The HFD for 6 months showed increased obesity, glucose intolerance, and insulin resistance in mice. Toll-like receptor 4 expression was found in vascular pericytes at the inner retina. Increased CD11b(+) and CD45(+) cells, phosphorylated NF-κB, interleukin-6, gp91phox, MDA, and γH2AX were observed in the retina of mice fed a HFD compared to CD counterparts. TLR4KO mice did not show the adverse effects of HFD. CONCLUSIONS: Our results indicate that HFD-induced macrophage/microglial cell activation and retinal impairment were reduced in the absence of TLR4. The findings suggest that TLR4 is implicated in the pathogenesis of retinal diseases caused by metabolic disorders.
Subject(s)
Diet, High-Fat/adverse effects , Macrophage Activation/physiology , Macrophages/metabolism , Microglia/metabolism , Obesity/metabolism , Oxidative Stress/physiology , Retina/metabolism , Retinal Diseases/metabolism , Toll-Like Receptor 4/metabolism , Animals , Fluorescent Antibody Technique , Glucose Intolerance , Inflammation/metabolism , Insulin Resistance , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Real-Time Polymerase Chain Reaction , Retinal Diseases/pathologyABSTRACT
We report a literature review and detailed evaluation of a rare case of posterior choroidal leiomyoma to emphasize the importance of differentiating this from other choroidal tumors. A 30-year-old male presented with variable blurred vision in his right eye secondary to a choroidal tumor. Clinical examinations were performed including fundus photography, optical coherence tomography, B scans, fluorescein and indocyanine green angiography, computed tomography, and magnetic resonance imaging. Preoperative examination revealed a suspected choroidal melanoma and enucleation was performed. However, a definitive diagnosis of choroidal leiomyoma was made following postoperative pathological light microscopy and immunohistochemical studies. Published case reports were collected and the common characteristics and distinctive features were compared with the current case. Posterior choroidal leiomyoma was summarized from the literature, and beneficial information for diagnosis and treatment was obtained. In conclusion, posterior choroidal leiomyoma is rare and should be differentiated from amelanotic melanomas. Despite the benign nature, an explanation regarding the rare incidence and difficult diagnosis of posterior choroidal leiomyoma must be provided to patients, prior to enucleation or detrimental treatment.
Subject(s)
Choroid Neoplasms/diagnosis , Choroid Neoplasms/therapy , Melanoma/diagnosis , Melanoma/therapy , Uveal Neoplasms/diagnosis , Uveal Neoplasms/therapy , Adult , Choroid Neoplasms/pathology , Humans , Leiomyoma/diagnosis , Leiomyoma/pathology , Leiomyoma/therapy , Male , Melanoma/pathology , Uveal Neoplasms/pathologyABSTRACT
Patients with diabetes mellitus have an increased risk of developing Alzheimer's disease. Amyloid-ß, a product of amyloid precursor protein, is associated with neuro-inflammation in patients with Alzheimer's diseases. The correlation between amyloid-beta and advanced glycation end products, which accumulate in tissue of diabetic patients, is not clear. The aims of this study were to determine the effect of advanced glycation end product on the expression of amyloid precursor protein/amyloid-beta and associated pro-inflammatory responses in retinal ganglion cell line RGC-5. Treatment with advanced glycation end product produced upregulation of amyloid precursor protein and increased secretion of amyloid-ß(1-40). Additionally, amyloid-ß(1-40) induced toll-like receptor 4-dependent phosphorylation of tyrosine in myeloid differentiation primary response gene (88). We found that N-[N-(3,5-Difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester, a γ-secretase inhibitor, reduced the secretion of amyloid-ß(1-40) and inhibited the advanced glycation end product-induced activation of myeloid differentiation primary response gene (88). Amyloid-ß(1-40) induced the activation of NF-κB and the expression of TNFα mRNA. Knockdown of toll-like receptor 4 inhibited the amyloid-ß(1-40)-induced phosphorylation of p65 in NF-κB. Additionally, the nuclear translocation of p65 and transcriptions of TNFα were inhibited by siRNA knockdown of receptor of advanced glycation end product or toll-like receptor 4. The advanced glycation end product-induced secretion of VEGF-A was also reduced by knockdown of toll-like receptor 4. Taken together, our data suggested that amyloid-ß(1-40) mediates the interaction between receptor of advanced glycation end product and toll-like receptor 4. Inhibition of the toll-like receptor 4 is an effective method for suppressing the amyloid-ß(1-40)-induced pro-inflammatory responses in RGC-5 cells.
Subject(s)
Amyloid beta-Peptides/physiology , Peptide Fragments/physiology , Receptor for Advanced Glycation End Products/physiology , Retinal Ganglion Cells/metabolism , Toll-Like Receptor 4/metabolism , Animals , Cell Line , Glycation End Products, Advanced/physiology , Myeloid Differentiation Factor 88/metabolism , NF-kappa B/metabolism , Rats , Retinal Ganglion Cells/immunology , Signal Transduction , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor A/metabolismABSTRACT
PURPOSE: Calcium signaling is an important intracellular pathway. Increased intracellular calcium is associated with cytokine regulation and inflammatory signals secretion. The purpose of this study is to understand the molecular mechanisms by which calcium signaling controls IL-8 activation in human RPE cells. METHODS: Fluorescence-based calcium imaging and different mutants of IL-8 plasmids were used in this study. The IL-8 promoter activation, gene expression, and secretion were detected by using luciferase reporter assay, quantitative real-time PCR (Q-PCR), and ELISA, respectively. In addition, pharmacological inhibitors and small interfering RNA (siRNA) were applied to clarify the mechanisms of IL-8 activation. RESULTS: Our study reported that intracellular calcium mobilization activated IL-8 gene expression and secretion. Application of pharmacological inhibitor BAY 11-7082, siRNA, and plasmids of the nuclear factor κ light chain enhancer of activated B cells (NF-κB) binding site, we identified that NF-κB is the main transcription factor involved in intracellular calcium mobilization-mediated IL-8 activation in human RPE cells. CONCLUSIONS: Collectively, our findings highlight the important role of intracellular calcium mobilization in the activation of IL-8. These findings may be helpful for the clinical applications in the age-related macular degeneration (AMD) prevention and treatment.
Subject(s)
Calcium/metabolism , Gene Expression Regulation , Interleukin-8/genetics , Macular Degeneration/metabolism , RNA/genetics , Retinal Pigment Epithelium/metabolism , Blotting, Western , Calcium Signaling , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Humans , Interleukin-8/biosynthesis , Macular Degeneration/pathology , RNA/metabolism , Real-Time Polymerase Chain Reaction , Retinal Pigment Epithelium/pathologyABSTRACT
AIMS: To explore the role of serum uric acid (SUA) concentration in diabetic retinopathy (DR) for patients with type 2 diabetes mellitus (T2DM). METHODS: A 3-year prospective study in 749 patients with T2DM and without proliferative diabetic retinopathy (PDR) was conducted at a medical center. Baseline SUA concentration and parameters of glycemic control, blood pressure, kidney disease, and lipid profiles were analyzed to determine their contribution to DR. RESULTS: Fundus examination showed that 184 patients (24.6%) had non-proliferative retinopathy and 565 (75.4%) without DR at baseline. After 3 years, increase in the severity of DR was recognized in 103 patients (13.8%), including 81 patients with newly developed DR. Patients with increase in severity of DR positively associated with duration of DM (11.9 vs. 9.4 years, p = 0.001), HbA1c (7.6 vs. 7.2%, p = 0.001), albuminuria (45.5 vs. 31.0%, p = 0.006), and SUA (6.47 vs. 5.87 mg/dl, p<0.001) than did those without change in DR stage. Cox regression showed that patients with SUA in the 3rd (5.9-6.9 mg/dl) and 4th (≥ 7.0mg/dl) quartiles had hazard ratios for DR worsening of 2.57 and 3.66 (95% C.I. 1.30-5.08 and 1.92-7.00) when compared with patients with SUA in the 1st quartile (<4.9 mg/dl). CONCLUSIONS: SUA concentration is associated with the increase in severity of DR over a 3-year period in patients with T2DM. Further study is required to define the exact role of SUA in DR.
Subject(s)
Diabetes Mellitus, Type 2/blood , Diabetic Retinopathy/blood , Uric Acid/blood , Adult , Aged , Aged, 80 and over , Blood Glucose/analysis , Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/epidemiology , Diabetic Retinopathy/epidemiology , Diabetic Retinopathy/pathology , Disease Progression , Female , Follow-Up Studies , Humans , Male , Middle Aged , Prospective Studies , Risk Factors , Severity of Illness Index , Taiwan/epidemiologyABSTRACT
BACKGROUND: In non-excitable cells, one major route for calcium entry is through store-operated calcium (SOC) channels in the plasma membrane. These channels are activated by the emptying of intracellular Ca²âº store. STIM1 and Orai1 are major regulators of SOC channels. In this study, we explored the functions of STIM1 and Orai1 in epidermal growth factor (EGF)-induced cell proliferation and migration in retinal pigment epithelial cells (ARPE-19 cell line). RESULTS: EGF triggers cell proliferation and migration in ARPE-19 cells. Cell proliferation and migration involve STIM1 and Orai1, as well as phosphorylation of extracellular signal-regulated protein kinase (ERK) 1/2, and Akt. Pharmacological inhibitors of SOC channels and siRNA of Orai1 and STIM1 suppress cell proliferation and migration. Pre-treatment of mitogen-activated protein kinase kinase (MEK) inhibitors and a phosphatidylinositol 3 kinases (PI3K) inhibitor attenuated cell proliferation and migration. However, inhibition of the SOC channels failed to prevent EGF-mediated ERK 1/2 and Akt phosphorylation. CONCLUSIONS: Our results showed that STIM1, Orai1, ERK 1/2, and Akt are key determinants of EGF-mediated cell growth in ARPE-19 cells. EGF is a potent growth molecule that has been linked to the development of PVR, and therefore, STIM1, Orai1, as well as the MEK/ERK 1/2 and PI3K/Akt pathways, might be potential therapeutic targets for drugs aimed at treating such disorders.
Subject(s)
Calcium Channels/metabolism , Epidermal Growth Factor/metabolism , Membrane Proteins/metabolism , Neoplasm Proteins/metabolism , Retinal Pigment Epithelium/metabolism , Calcium/metabolism , Calcium Channels/genetics , Cell Line , Cell Membrane/metabolism , Cell Movement , Cell Proliferation , Humans , Membrane Proteins/genetics , Mitogen-Activated Protein Kinase Kinases/metabolism , Neoplasm Proteins/genetics , ORAI1 Protein , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , Stromal Interaction Molecule 1ABSTRACT
PURPOSE: High-mobility group box 1 protein (HMGB1) has been reported to be a potent proangiogenic factor induced by inflammatory stress. In this study, we explore the role of HMGB1 in advanced glycation end products (AGEs)-induced vascular endothelial growth factor A (VEGF-A) production in rat retinal ganglion cell line 5 (RGC-5) cells. METHODS: The VEGF-A protein and mRNA levels in conditioned medium of RGC-5 cells incubated with AGE-modified BSA (AGE-BSA) were examined with real-time PCR and enzyme-linked immunosorbent assay (ELISA), and BSA-treated cells were used as controls. The expression of HMGB1, c-Jun N-terminal kinase (JNK), extracellular-signal-regulated kinase (ERK), and p38 mitogen-activated protein kinase (p38 MAPK) was assessed with immunofluorescence and western blot analysis. Reactive oxidative species (ROS) were detected with flow cytometry measurements of peroxide-dependent oxidation of 2'-7'-dichlorofluorescein-diacetate (DCFH-DA). N-Acetyl-L-cysteine (NAC), glycyrrhizin (GZ), and SP600125 were used to block ROS, HMGB1, and JNK, respectively. RESULTS: Compared with the BSA controls, the RGC-5 cells incubated with AGE-BSA showed a dose- and time-dependent increase in VEGF-A mRNA and VEGF-A protein secretion in the supernatant, with the highest levels achieved at 24 h. AGE-BSA stimulated a significant release of HMGB1 in the supernatant and a significant increase of intracellular ROS production at 3 h. NAC blocked HMGB1 production in a dose-dependent manner. Blocking with GZ, NAC, and JNK significantly suppressed AGE-induced VEGF-A production. CONCLUSIONS: HMGB1 is implicated in the production of VEGF-A in retinal ganglion cell line-5 (RGC-5). Blocking HMGB1, ROS, or the JNK pathway may attenuate VEGF-A production, suggesting HMGB1 and related signaling molecules play a role in diabetic retinopathy.
Subject(s)
Glycation End Products, Advanced/pharmacology , HMGB1 Protein/biosynthesis , RNA, Messenger/biosynthesis , Retinal Ganglion Cells/metabolism , Vascular Endothelial Growth Factor A/biosynthesis , Acetylcysteine/pharmacology , Animals , Anthracenes/pharmacology , Cattle , Cell Line , Culture Media, Conditioned/pharmacology , Dose-Response Relationship, Drug , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Extracellular Signal-Regulated MAP Kinases/metabolism , Glycyrrhizic Acid/pharmacology , HMGB1 Protein/metabolism , MAP Kinase Kinase 4/antagonists & inhibitors , MAP Kinase Kinase 4/metabolism , Rats , Reactive Oxygen Species/antagonists & inhibitors , Reactive Oxygen Species/metabolism , Retinal Ganglion Cells/cytology , Retinal Ganglion Cells/drug effects , Serum Albumin, Bovine/pharmacology , Signal Transduction/drug effects , Vascular Endothelial Growth Factor A/metabolism , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Cyclooxygenase-2 (COX-2) is an inducible isoform of the enzyme responsible for the synthesis of several inflammatory mediators. In a search for phytochemicals with anti-inflammatory activity, the COX-2 inhibitory activity of 15 typical Annonaceous acetogenins was examined. Isodesacetyluvaricin (1), from the Formosan tropical fruit tree Annona glabra, exhibited the most potent activity. Reverse transcription PCR was used to test the effect of 1 on epidermal growth factor-stimulated expression of COX-2 in cultures of A431 human epidermoid carcinoma cells. Three hours after exposure to 1 (5 µM), A431 cells had barely detectable levels of COX-2 mRNA. A corresponding but smaller decline in the COX-2 protein appeared on using Western blots. Lipopolysaccharide-stimulated expression of COX-2 in Raw 264.7 mouse leukemic monocyte-macrophages showed a similar decrease. Luciferase assays revealed that cells exposed to 1 had reduced activities of two COX-2 promoter-transcription factors: cAMP response element-binding factor and nuclear factor of activated T-cells. Compound 1 did not affect cell proliferation, as measured by a colorimetric assay, or intracellular store-operated calcium influx, as determined by fluorescence imaging. Thus, 1 may serve as a lead compound for targeting inflammatory diseases as well as angiogenesis and cancer metastasis.
Subject(s)
Acetogenins/pharmacology , Cyclooxygenase 2/metabolism , Acetogenins/chemistry , Annona/chemistry , Calcium/analysis , Cyclooxygenase 1/drug effects , Cyclooxygenase 1/genetics , Cyclooxygenase 1/metabolism , Cyclooxygenase 2/drug effects , Cyclooxygenase 2/genetics , Cyclooxygenase 2 Inhibitors/pharmacology , Dose-Response Relationship, Drug , Epidermal Growth Factor/pharmacology , Humans , Indomethacin/pharmacology , Molecular Structure , Seeds/chemistry , TaiwanABSTRACT
BACKGROUND: Retinoblastomas are the most common malignant intraocular tumor of childhood. We describe the survival outcomes and prognostic factors of patients with a retinoblastoma receiving primary treatment at our hospital over the last 20 years. METHODS: A retrospective series study of 30 retinoblastoma cases treated from January 1, 1987 to August 31, 2006 was conducted from a review of medical records and histopathological sections. Variables, including age at onset, laterality, treatment modalities, treatment delay, and optic nerve invasion, were analyzed to elucidate the prognostic factors associated with cumulative survival. RESULTS: Most of the cases had an advanced retinoblastoma, and 23 patients received enucleation treatment. The average period of delay for treatment was 5.37 months after discovery of the disease. The overall cumulative survival rate was 83.08%. Patients with optic nerve invasion had a significantly lower survival rate (60.0%) than those without optic nerve involvement (94.75%). Treatment delay in excess of 6 months was correlated with tumor invasion of the optic nerve. CONCLUSIONS: Tumor invasion of the optic nerve is the most significant prognostic factor for surviving a retinoblastoma. Delayed treatment increases the risk of optic nerve invasion. Parental awareness of both the risk of this consequence and the significance of early treatment is vital to achieving improved survival rates.
Subject(s)
Retinal Neoplasms/surgery , Retinoblastoma/surgery , Child, Preschool , Female , Humans , Infant , Male , Retinal Neoplasms/mortality , Retinoblastoma/mortality , Retrospective Studies , Taiwan/epidemiologySubject(s)
Burkholderia pseudomallei/isolation & purification , Endophthalmitis/microbiology , Eye Infections, Bacterial/microbiology , Melioidosis/microbiology , Shock, Septic/microbiology , Aged , Anti-Bacterial Agents/therapeutic use , Drug Therapy, Combination , Endophthalmitis/diagnosis , Endophthalmitis/drug therapy , Eye Infections, Bacterial/diagnosis , Eye Infections, Bacterial/drug therapy , Fatal Outcome , Humans , Male , Melioidosis/diagnosis , Melioidosis/drug therapy , Shock, Septic/diagnosis , Shock, Septic/drug therapy , Vitreous Body/microbiologyABSTRACT
AIM: The aim of this study was to evaluate the efficacy of 0.05% atropine solution for controlling myopia progression in school-aged children. RESULTS: This retrospective, case-control study enrolled myopic school-aged children who had presented at Kaohsiung Chang Gung Memorial Hospital (Kaohsiung, Taiwan) from 2001 to 2004. A group of 57 children (30 boys, 27 girls; 6-12 years of age) with regular follow-up was divided into a subgroup of 21 children (12 boys, 9 girls) who received atropine eyedrops (0.05%) every evening, and a subgroup of 36 children (18 boys, 18 girls), who remained untreated, served as controls. The changes in refractive status of 114 eyes in 57 children were collected and compared for patients treated with 0.05% atropine eyedrop and those without medical control. The initial spherical equivalent of refractive status range was between -0.5 and -5.5 D. Mean myopia progression for the group of patients treated with 0.05% atropine eyedrop (n = 21) was -0.28 +/- 0.26 D/year, significantly lower than that of the control group of -0.75 +/- 0.35 D/year (36 patients; P < 0.001). The 0.05% atropine group had a significant lower ratio of uncontrolled myopia, that progressed greater than -0.50 D in 1 year, relative to the controls (16.7% versus 77.8%; P < 0.001). CONCLUSIONS: The results of this study demonstrate that, with regular instillation, topical 0.05% atropine is an effective agent for controlling myopia progression in a majority of school-aged children for at least a period of 1 year.