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Full-Text Articles in Physical Sciences and Mathematics

Iron-Dithiothreitol Dependent Production Of 8-Hydroxy-2'-Deoxyguanosine In Dna, David Allan Cook May 1995

Iron-Dithiothreitol Dependent Production Of 8-Hydroxy-2'-Deoxyguanosine In Dna, David Allan Cook

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Oxidative damage to DNA in a metal-thiol system was studied. Calf thymus DNA was incubated in an iron(IIl)-dithiothreitol system prepared in various buffers. Damage was measured by monitoring the production of 8-hydroxy-2'-deoxyguanosine. DNA oxidation was proportional to the concentration of dithiothreitol (DTT) in HEPES and tris(hydroxymethyl)aminomethane (Tris) buffers while it was only roughly related to DTT concentration in sodium chloride. Incubation in potassium phosphate buffer produced no damage. The results indicate that hydroxyl radicals may be generated by the iron-OTT system in HEPES, Tris, and sodium chloride, while in the phosphate buffer an iron phosphate complex may be formed which …


Detection Of Dityrosine In Apoferritin, Stephen E. Bialkowski Jan 1995

Detection Of Dityrosine In Apoferritin, Stephen E. Bialkowski

Stephen E. Bialkowski

Laser-excited fluorescence spectroscopy was adopted for dityrosine detection in horse spleen apoferritin. Dityrosine was identified in horse spleen apoferritin. Fractionation of acid-hydrolyzed horse spleen apoferritin showed fluorescence attributed to dityrosine, and other compounds, excited by 325-nm laser radiation. The other fluorescent compounds resulted in a broad fluorescence emission spectrum of horse spleen apoferritin. The broad emission band overlapped with the fluorescence emission spectrum of pure dityrosine. Fractionation of horse spleen apoferritin acid hydrolysate prior to laser-excited fluorescence detection was necessary for dityrosine quantitative analysis. The concentration of dityrosine in horse spleen apoferritin was 2.5% (mol/mol), estimated from the method of …


Laser Excited Fluorescence Of Dityrosine, Sahar F. Mahmoud, Stephen E. Bialkowski Jan 1995

Laser Excited Fluorescence Of Dityrosine, Sahar F. Mahmoud, Stephen E. Bialkowski

Stephen E. Bialkowski

In this research, laser-excited fluorescence was examined for sensitive detection of aqueous dityrosine. Samples were excited with a 6.3-mW, 325-nm helium-cadmium laser focused into a small volume-fluorescence cell with a 10-cm lens. The resulting fluorescence emission was collected perpendicular to the excitation and detected with two different schemes. An optical bandpass filter was used with a photomultiplier tube for sensitive quantitative measurement, while a photodiode array detector was used in conjunction with a spectrograph for qualitative characterization of fluorescence emission spectra. Dityrosine detection on the order of 2 × 10-11 M was obtained with the use of the photomultiplier …