Sodium Fluorescein Staining of the Cornea for the Diagnosis of Dry Eye: A Comparison of Three Eye Solutions

The purpose of this study was to identify which of the eye solutions is best for sodium fluorescein staining of the cornea to diagnose dry eye disease. The study included 173 eyes with suspected or known dry eye disease. The eyes were stained sequentially with sodium fluorescein and each of the following four conditions: balanced salt solution (BSS); BSS and cyclosporine 0.05% emulsion; BSS and lipids containing omega-3; and BSS, cyclosporine 0.05% emulsion, and lipids containing omega-3. Our results showed that compared to BSS alone, artificial tears with cyclosporine 0.05% emulsion and lipids containing omega-3 remain in the cornea for longer periods, thus allowing the clinician to evaluate tear break-up time and visualize corneal punctate erosions

IGF-I Activation of the AKT Pathway Is Impaired in Visceral But Not Subcutaneous Preadipocytes from Obese Subjects

Obesity morbidity is associated with excess visceral adiposity, whereas sc adipose tissue is much less metabolically hazardous. Human abdominal sc preadipocytes have greater capacity for proliferation, differentiation, and survival than omental preadipocytes. IGF-I is a critical mediator of preadipocyte proliferation, differentiation, and survival through multiple signaling pathways. We investigated IGF-I action in primary cultures of human preadipocytes isolated from sc and omental adipose tissue of obese subjects. IGF-I-stimulated DNA synthesis was significantly lower in omental compared with sc preadipocytes. IGF-I phosphorylation of the IGF-I receptor and the ERK pathway was comparable in sc and omental cells. However, omental preadipocytes had decreased insulin receptor substrate (IRS)-1 protein associated with increased IRS-1-serine636/639 phosphorylation and degradation. IGF-I-stimulated phosphorylation of AKT on serine473 but not threonine308 was decreased in omental cells, and activation of downstream targets, including S6Kinase, glycogen synthase kinase-3, and Forkhead box O1 was also impaired. CyclinD1 abundance was decreased in omental cells due to increased degradation. Over-expression of IRS-1 by lentivirus in omental preadipocytes increased IGF-I-stimulated AKT-serine473 phosphorylation. The mammalian target of rapamycin (mTOR)-Rictor complex regulates phosphorylation of AKT-serine473 in 3T3-L1 adipocytes, but knockdown of Rictor by lentivirus-delivered short hairpin RNA in sc preadipocytes did not affect AKT-serine473 phosphorylation by IGF-I. These data reveal an intrinsic defect in IGF-I activation of the AKT pathway in omental preadipocytes from obese subjects that involves IRS-1 but probably not mTOR-Rictor complex. We conclude that impaired cell cycle regulation by AKT contributes to the distinct growth phenotype of preadipocytes in visceral fat of obese subjects