To elucidate the mechanism of increased intraocular pressure in primary open-angle glaucoma (POAG), the protein profiles of aqueous humor obtained from POAG patients were compared with those of cataract patients as a control group. Aqueous humor proteins were analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and detected by the ultrasensitive silver staining technique. In 79% of the samples taken from POAG patients, protein bands of 140,000 or 160,000 daltons were stained, but none were stained from cataract patients. The presence of these protein bands revealed statistically significant differences between the two groups. Protein bands of 140,000 or 160,000 daltons were evenly visible at all ages in POAG patients, and the positivity of bands had no correlation with sex or initial intraocular pressure level. It is possible that the ultrastructural changes of the aqueous outflow pathway in POAG may be related to the changes in the aqueous protein, presence of 140,000 or 160,000 daltons protein bands.
Adult ; Aged ; Aqueous Humor/*metabolism ; Cataract/metabolism ; Electrophoresis, Polyacrylamide Gel ; Eye Proteins/*metabolism ; Female ; Glaucoma, Open-Angle/*metabolism ; Humans ; Male ; Middle Aged ; Molecular Weight

To understand the pathogenesis of proliferative vitreoretinal membrane formation which occurs in proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR), etc., accurate identification of the cellular components of the membrane is needed. This study was performed to identify cellular components of the membranes by means of immunohistochemical technique. 11 proliferative vitreoretinal membranes which were surgically obtained from 7 eyes with PVR and 4 eyes with PDR were stained with monoclonal antibodies against cytokeratin, glial fibrillary acidic protein (GFAP), or vimentin using immunoperoxidase technique (ABC method). In the PVR membranes, mean cell positivities for cytokeratin, GFAP and vimentin were 48%, 1% and 92%, respectively and in the PDR membranes, 0%, 5% and 93%, respectively. The above results suggest that retinal pigment epithelial cells and fibroblasts are major cellular components of PVR membranes, and that mesenchymal cells are major cellular components and glial cells are minor cellular components of PDR membranes.
Adolescent ; Adult ; Antibodies, Monoclonal ; Cell Membrane/metabolism/pathology ; Diabetic Retinopathy/metabolism/pathology ; Eye Diseases/metabolism/pathology ; Female ; Humans ; Immunoenzyme Techniques ; Intermediate Filament Proteins/*analysis ; Male ; Middle Aged ; Retinal Diseases/metabolism/*pathology ; Vitreous Body/metabolism/*pathology

PURPOSE: To describe clinical findings in a Korean family with Axenfeld-Rieger syndrome. METHODS: A retrospective review of clinical data about patients with diagnosed Axenfeld-Rieger syndrome. Five affected members of the family underwent a complete ophthalmologic examination. We screened the forkhead box C1 gene and the pituitary homeobox 2 gene in patients. Peripheral blood leukocytes and buccal mucosal epithelial cells were obtained from seven members of a family with Axenfeld-Rieger syndrome. DNA was extracted and amplified by polymerase chain reaction, followed by direct sequencing. RESULTS: The affected members showed iris hypoplasia, iridocorneal adhesions, posterior embryotoxon, and advanced glaucoma in three generation. None had systemic anomalies. Two mutations including c.1362_1364insCGG and c.1142_1144insGGC were identified in forkhead box C1 in four affected family members. CONCLUSIONS: This study may help to understand clinical findings and prognosis for patients with Axenfeld-Rieger syndrome.
Aged, 80 and over ; Anterior Eye Segment/*abnormalities/metabolism ; DNA/*genetics ; DNA Mutational Analysis ; Eye Abnormalities/diagnosis/*genetics/metabolism ; Female ; Forkhead Transcription Factors/*genetics/metabolism ; Genetic Testing ; Homeodomain Proteins/*genetics/metabolism ; Humans ; Male ; Middle Aged ; *Mutation ; Pedigree ; Retrospective Studies ; Transcription Factors/*genetics/metabolism ; Young Adult

PURPOSE: This study was performed to describe the ultrastructure of stromal nerve fibers in central, mid-peripheral, and peripheral parts of the human cornea by flat serial corneal section. METHODS: Seven samples from fresh normal cornea, derived from eyes with retinoblastoma and eyes from eye bank, were processed for transmission electron microscopic examinations. Flat serial sections reaching from mid-epithelium to the anterior stroma were observed. RESULTS: The myelinated and unmyelinated nerve fibers are alternately arranged and run parallel to the stromal collagen fibers at the periphery of the cornea. The main difference between the limbal and the central cornea is the presence of myelinated nerve fibers in the anterior peripheral stroma. The diameter of the unmyelinated nerve fiber measures between 0.25 and 0.63 micrometer in size. The corneal nerve fibers contain vesicles, mitochondria, and glycogen particles. The peripheral nerve fibers contain both clear and dense vesicles. The nerve fiber is separated by an interval of 0.3 micrometer from the cytoplasmic wall of keratocyte at the center of the cornea. CONCLUSIONS: The majority of the corneal nerve fibers can be classified as C-fibers due to their size. The presence of both clear and dense vesicles within the cytoplasm of the periphery of the cornea suggest that a small portion in the peripheral corneal nerve may be originated from the sympathetic nervous system. A close vicinity between the nerve fibers and keratocyte supports that nerve fibers might modulate the release of growth factors in the regulation of stromal and epithelial metabolism.
Collagen ; Cornea ; Cytoplasm ; Eye Banks ; Glycogen ; Humans ; Intercellular Signaling Peptides and Proteins ; Metabolism ; Mitochondria ; Myelin Sheath ; Nerve Fibers ; Nerve Fibers, Myelinated ; Nerve Fibers, Unmyelinated ; Peripheral Nerves ; Retinoblastoma ; Sympathetic Nervous System

In this study, we investigated whether retinal soluble proteins, such as interphotoreceptor retinoid-binding protein(IRBP), play a role in the induction of nitric oxide by macrophages in vitro. Cells from the murine macrophage cell line RAW 264.7 and rat and rabbit peritoneal macrophages were incubated in the presence of retinal soluble protein. The nitrite level in the cultured supernatant was evaluated for nitric oxide production using the Griess reaction. IRBP induced significant, dose-dependent nitrite production in both RAW 264.7 and rat peritoneal macrophages. Induction of inducible nitric oxide synthase (iNOS) by retinal proteins was inhibited by the iNOS-specific inhibitor, aminoguanidine, and the tyrosine inhibitor, genistein. These results show that soluble retinal proteins significantly induce nitric acid production by macrophages. Increased production of reactive oxygen species by macrophages in the presence of this soluble retinal protein in vivo may accelerate photoreceptor degeneration in uveitis.
Animal ; Cell Line ; Comparative Study ; Enzyme Inhibitors/pharmacology ; Eye Proteins/pharmacology* ; Guanidines/pharmacology ; Macrophages, Peritoneal/metabolism ; Macrophages, Peritoneal/drug effects* ; Macrophages, Peritoneal/cytology ; Nitric Oxide/biosynthesis* ; Nitric-Oxide Synthase/metabolism ; Nitric-Oxide Synthase/antagonists & inhibitors ; Rabbits ; Rats ; Rats, Inbred Lew ; Retinol-Binding Proteins/pharmacology*

PURPOSE: Hepatocyte growth factor (HGF) and its receptor (HGFR/c-Met) regulate motility, mitogenesis, and morphogenesis in a cell type-dependent fashion. We report the role of HGF and c-Met on stress-induced ARPE-19 human retinal pigment epithelial (RPE) cells in this study. METHODS: The cells were cultured either with or without serum. Southern and Western blot analyses were done to determine the expression patterns of HGF/c-Met in serum-starved ARPE-19 cells. The cell proliferation pattern in serum-starved condition was analyzed using MTS assay. Inhibition level of cell proliferation was analyzed using a neutralizing monoclonal antibody against c-Met (2 microgram/ml). RESULTS: Abnormal cell proliferation and scattering of ARPE-19 cells was observed under serum starvation. HGF/c-Met were expressed in serum-starved ARPE-19 cells. ARPE-19 cell proliferation was also enhanced with recombinant HGF treatment. Neutralization against c-Met inhibited the proliferation of serum-deprived ARPE-19 by 64.5% (n=9, S.D. 5.5%). Serum starvation appears to induce epithelial-mesenchymal transition of ARPE-19 cells, resulting in scatter, and the expression of alpha-smooth muscle actin (alpha-SMA), a marker for fibrosis. CONCLUSIONS: In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.
Blotting, Southern ; Blotting, Western ; Cell Movement/physiology ; Cell Proliferation ; Cells, Cultured ; Culture Media, Serum-Free ; *Gene Expression ; Hepatocyte Growth Factor/biosynthesis/*genetics ; Humans ; Mitosis/physiology ; Pigment Epithelium of Eye/cytology/*metabolism ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-met/biosynthesis/*genetics ; RNA/*genetics