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Purpose To evaluate feasibility and repeatability of measures for ocular sun exposure and conjunctival ultraviolet autofluorescence (UVAF), and to test for relationships between the outcomes. Methods Fifty volunteers were seen for 2 visits 14±2 days apart. Ocular sun exposure was estimated over a two-week time period using questionnaires that quantified time outdoors and ocular protection habits. Conjunctival UVAF was imaged using a Nikon D7000 camera system equipped with appropriate flash and filter system; image analysis was done using ImageJ software. Repeatability estimates were made using Bland-Altman plots with mean differences and 95% limits of agreement calculated. Non-normally distributed data was transformed by either log10 or square root methods. Linear regression was conducted to evaluate relationships between measures. Results Mean (±SD) values for ocular sun exposure and conjunctival UVAF were 8.86 (±11.97) hours and 9.15 (±9.47) mm2, respectively. Repeatability was found to be acceptable for both ocular sun exposure and conjunctival UVAF. Univariate linear regression showed outdoor occupation to be a predictor of higher ocular sun exposure; outdoor occupation and winter season of collection both predicted higher total UVAF. Furthermore, increased portion of day spent outdoors while working was associated with increased total conjunctival UVAF. Conclusions We demonstrate feasibility and repeatability of estimating ocular sun exposure using a previously unreported method and for conjunctival UVAF in a group of subjects residing in Ohio. Seasonal temperature variation may have influenced time outdoors and ultimately calculation of ocular sun exposure. As winter season of collection and outdoor occupation both predicted higher total UVAF, our data suggests that ocular sun exposure is associated with conjunctival UVAF and possibly, that UVAF remains for at least several months following sun exposure.

Purpose To compare various meibum collection methods and extraction techniques. Methods Sixty subjects, all successful contact lens wearers, were seen on two visits. Meibum was collected from the lower lid of the right eye with a glass microcapillary tube, and with a Dacron swab, cytology microbrush or spatula from the left eye. Extraction with 2:1 chloroform:methanol was done either immediately or after data collection was complete. Individual samples were divided into four equal aliquots for analysis of total lipids, cholesterol and inorganic phosphates via assay based techniques. Effects of collection method, extraction, and dry eye status were examined using repeated measures ANOVA and logistic regression. Results Total lipids showed significance of collection device (p<0.0001), but not for extraction technique (p=0.13) or dry eye status (p=0.97). Dacron swab collection was associated with more total lipid on average than each other collection device (p<0.0001). The cholesterol assay showed significance of collection device (p<0.0001) and extraction technique (p=0.0002), but not dry eye status (p=0.55). Spatula collection was associated with more cholesterol on average than each other collection device (p<0.0001). For inorganic phosphates, immediate extraction (p<0.0001), cytology microbrush collection (p<0.0001), and non-dry eye status (p=0.03) were associated with the greater likelihood of detection. Conclusions Dacron swab collection was associated with the highest average amount of total lipid detected, whereas spatula collection and immediate extraction was associated with the highest average amount of cholesterol detected. Cytology microbrush collection with immediate extraction on non-dry eye subjects was associated with the highest probability of detection of inorganic phosphates.

PurposeWe evaluate feasibility and repeatability of measures for lipid peroxidation and DNA oxidation in human tears, as well as relationships between outcome variables, and compared our findings to previously reported methods of evaluation for ocular sun exposure.MethodsA total of 50 volunteers were seen for 2 visits 14 ± 2 days apart. Tear samples were collected from the inferior tear meniscus using a glass microcapillary tube. Oxidative stress biomarkers were quantified using enzyme-linked immunosorbent assay (ELISA): lipid peroxidation by measurement of hexanoyl-lysine (HEL) expression; DNA oxidation by measurement of 8-oxo-2′-deoxyguinosone (8OHdG) expression. Descriptive statistics were generated. Repeatability estimates were made using Bland-Altman plots with mean differences and 95% limits of agreement were calculated. Linear regression was conducted to evaluate relationships between measures.ResultsMean (±SD) values for tear HEL and 8OHdG expression were 17368.02 (±9878.42) nmol/L and 66.13 (±19.99) ng/mL, respectively. Repeatability was found to be acceptable for both HEL and 8OHdG expression. Univariate linear regression supported tear 8OHdG expression and spring season of collection to be predictors of higher tear HEL expression; tear HEL expression was confirmed as a predictor of higher tear 8OHdG expression.ConclusionsWe demonstrate feasibility and repeatability of estimating previously unreported tear 8OHdG expression. Seasonal temperature variation and other factors may influence tear lipid peroxidation. Support is demonstrated to suggest lipid damage and DNA damage occur concurrently on the human ocular surface.

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