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A Midelfart, Ø Risa, O Sæther, S Løfgren, PG Søderberg, J Krane; Metabolic changes in rat lens after in vivo exposure to ultraviolet radiation measured by Magic Angle Spinning NMR spectroscopy . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3571.
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Purpose:Application of High Resolution Magic Angle Spinning (HRMAS) NMR spectroscopy as a non-destructive analytical tool for monitoring metabolic profile of rat lenses exposed to different UV radiation (UVR) doses. Methods:Sprague Dawley rats were exposed to UVR doses of 2.0kJ/m2, 5.0kJ/m2 or 7.5kJ/m2 on one eye, the other serving as a control. One week after exposure the rats were killed and forward light scattering was measured in the isolated lenses to quatify opacity. The lenses were then frozen.The metabolic profile was analyzed directly with proton HR MAS NMR spectroscopy in a spectrometer operating at 600 MHz. Results: After UVR exposure, the forward light scattering of exposed lenses differed significantly from that of control lenses. The obtained proton HR MAS NMR spectra were of high quality, comparable with spectra from lens extracts. A decrease of a large number of metabolites such as valine, succinate, hypotaurine, taurine, myo-inositol, choline, tyrosine and glutathione was detected in the exposed lenses. However,alanin concentration was increased, while lactate and glutamate levels remained unchanged.The detected metabolic changes were not significantly related to the applied UVR dosis. Conclusion:Proton HR MAS NMR spectroscopy was found to be a valuable, non-destructive method for direct monitoring of metabolic changes occuring within the lens after UVR exposure. Concentration level of a number of water soluble substances in the lens could be simultaneously followed expanding our knowledge of the UVR induced cataract.
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