Marked reduction of alcohol dehydrogenase in keratoconus corneal fibroblasts.

PURPOSE: To identify differentially expressed genes in keratoconus (KC) corneal fibroblasts. METHODS: Stromal keratocytes (having a fibroblast morphology) from KC keratoplasty specimens and eye bank donor corneas were isolated and expanded using a serum containing medium. RNA was isolated from three KC fibroblast cultures and five eye bank donor cornea fibroblast cultures. The targets from the cultured fibroblasts were hybridized to the Affymetrix U133 Plus 2.0 microarrays. Western blot analyses of cell lysates were performed to examine protein levels of interest in the two groups. Protein levels of select differentially expressed genes were further examined by immunohistochemistry. Keratocyte staining of archived KC keratoplasty specimens were graded using a 0 to 3+ scale and compared to five archived whole globes having normal corneas as well as to 10 Fuchs' dystrophy keratoplasty specimens. RESULTS: Microarray analysis revealed up to a 212 fold reduction in the mRNA levels of alcohol dehydrogenase (class 1) beta polypeptide (ADH1B) in KC fibroblasts (p=0.04). Decreased alcohol dehydrogenase in KC fibroblasts was confirmed by western blot analysis of early passage primary keratocyte cell lysates. Immunohistochemistry using a monoclonal mouse immunoglobulin G (IgG) against human liver alcohol dehydrogenase revealed a dramatic difference in protein staining in the keratocytes of the KC group compared to the normal cornea group. Immunohistochemistry also showed decreased immunostaining against alcohol dehydrogenase in the KC stromal sections compared to those obtained from Fuchs' endothelial corneal dystrophy samples. CONCLUSIONS: Decreased alcohol dehydrogenase in KC corneal fibroblasts represents a strong marker and possible mediator of keratoconus.

Postprandial transient visual loss. A symptom of critical carotid stenosis.

PURPOSE: The authors report the association of transient visual loss after eating meals with severe carotid occlusive disease, and propose a hypothesis for its pathophysiology. METHODS: Description of clinical history and examination, radiologic and other studies in two patients with postprandial transient visual loss, and review of the literature for three related cases. RESULTS: Two women, 59 and 65 years of age, presented with splotchy visual loss lasting up to more than 1 hour in the left and both eyes, respectively. In both patients, the visual loss was precipitated by eating a meal. Radiologic investigations revealed 90% stenosis of the left internal carotid artery in the first patient and occlusion of the right internal carotid artery and 40% to 70% stenosis of the left internal carotid artery in the second patient. CONCLUSIONS: Visual loss after eating a meal may result from hypoperfusion of the retinal and choroidal circulations and is suggestive of severe carotid occlusive disease.