Autophagy and Exocytosis of Lipofuscin Into the Basolateral Extracellular Space of Human Retinal Pigment Epithelium From Fetal Development to Adolescence.

Purpose: To undertake the first ultrastructural characterization of human retinal pigment epithelial (RPE) differentiation from fetal development to adolescence. Methods: Ten fetal eyes and three eyes aged six, nine, and 17 years were examined in the temporal retina adjacent to the optic nerve head by transmission electron microscopy. The area, number, and distribution of RPE organelles were quantified and interpreted within the context of adjacent photoreceptors, Bruch's membrane, and choriocapillaris maturation. Results: Between eight to 12 weeks' gestation (WG), pseudostratified columnar epithelia with apical tight junctions differentiate to a simple cuboidal epithelium with random distribution of melanosomes and mitochondria. Between 12 to 26 WG, cells enlarge and show long apical microvilli and apicolateral junctional complexes. Coinciding with eye opening at 26 WG, melanosomes migrate apically whereas mitochondria distribute to perinuclear regions, with the first appearance of phagosomes, complex granules, and basolateral extracellular space (BES) formation. Significantly, autophagy and heterophagy, as evidenced by organelle recycling, and the gold standard of ultrastructural evidence for autophagy of double-membrane autophagosomes and mitophagosomes were evident from 32 WG, followed by basal infoldings of RPE cell membrane at 36 WG. Lipofuscin formation and deposition into the BES evident at six years increased at 17 years. Conclusions: We provide compelling ultrastructural evidence that heterophagy and autophagy begins in the third trimester of human fetal development and that deposition of cellular byproducts into the extracellular space of RPE takes place via exocytosis. Transplanted RPE cells must also demonstrate the capacity to subserve autophagic and heterophagic functions for effective disease mitigation.

Role of percent peripheral tissue ablated on refractive outcomes following hyperopic LASIK.

OBJECTIVES: To determine the effect of hyperopic laser in situ keratomileusis (H-LASIK) on corneal integrity, by investigating relationships between proportionate corneal tissue ablated and refractive outcomes at 3 months. METHODS: 18 eyes of 18 subjects treated with H-LASIK by Technolas 217c Excimer Laser were included in the study. Orbscan II Topography System was used to determine corneal volume and pachymetry 3mm temporally (3T). The volume of corneal tissue ablated was determined from the laser nomogram. Univariate associations between age, treatment, corneal volume, overall proportion of tissue removed, proportion of tissue removed at 3T, residual bed thickness at 3T and refractive outcomes 3 months post-LASIK were examined and independent factors associated with refractive outcomes determined using linear regression models. RESULTS: At 3 months post-LASIK, the mean difference to expected refractive outcome was -0.20 ± 0.64 (Range -2.00 to +1.00). In univariate analysis, difference to expected refractive outcome was associated with proportion of tissue removed at 3T (P<0.01, r = -0.605) and total number of pulses (P< 0.05, r = -0.574). In multivariable analysis, difference to expected refractive outcome was associated with the proportion of tissue removed at 3T only. CONCLUSION: Subjects undergoing H-LASIK, may present as either over or under-corrected at 3 months. The proportion of tissue removed at 3T was the single significant determinant of this outcome, suggesting unexpected biomechanical alterations resulting in corneal steepening. Future hyperopic LASIK procedures could consider proportionate volume of corneal tissue removed at 3T in addition to laser nomograms to achieve improved refractive outcomes.

Exercise-heat acclimation in young and older trained cyclists.

OBJECTIVES: The purpose of this study was to investigate the effect of age on the capacity to acclimatise to exercise-heat stress. This study hypothesised that age would not affect body temperature and heat loss effector responses to short-term exercise-heat acclimation in trained subjects. DESIGN: Seven young subjects (19-32 years) were matched with 7 older subjects (50-63 years). Subjects were highly trained but not specifically heat acclimated when they exercised for 60 min at 70%VO2max in hot-dry (35 °C, 40%RH) and thermoneutral (20 °C, 40%RH) conditions, pre and post 6 days of exercise-heat acclimation (70%VO2max, 35 °C, 40%RH). METHODS: Rectal temperature (Tr), skin temperature (Tsk), heart rate (HR), cutaneous vascular conductance (CVC) and whole body sweat loss (Msw) were measured during each testing session and Tr and HR were measured during each acclimation session. RESULTS: Tr, Tsk, %HRmax, CVC and Msw were similar across age groups both pre and post heat acclimation. Following heat acclimation relative decreases and increases in Tr and Msw, respectively, were similar in both subject groups. There was a significant reduction in heart rate (%HRmax) and increase in final CVC following the acclimation programme in the young group (all p < 0.05) but not the older group. CONCLUSIONS: When comparing young and older well trained adults we found age affected the cardiovascular adaptation but not body temperature or whole body sweat loss to exercise-heat acclimation. These data suggest age does not affect the capacity to acclimatise to exercise-heat stress in highly trained adults undergoing short-term heat acclimation.