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1991). The activity of succinate dehydrogenase (SDH), a mitochondrial click here enzyme in the Krebs cycle, was measured as an index of the oxidative capacity of the muscles. Sternohyoid and diaphragm muscle samples from animals in all the experimental groups were processed together in identical conditions. Succinate dehydrogenase activity was determined using an incubation solution as described by Fratacci et al. (1996b). Briefly, the solution contained the following: sodium succinate, 50 mm; nitroblue tetrazolium chloride, 1.5 mm; 1-methoxy-phenazine methosulfate (electron carrier), 0.2 mm; sodium azide (electron transport chain inhibitor), 0.1 mm; and distilled water (pH 7.4). The solution was heated to 37��C, and slides were incubated in this solution for 10 min. Muscle mitochondrial glycerol-3-phosphate dehydrogenase (GPDH) activity was separately quantified as an index of the glycolytic capacity of the muscles from all experimental groups. The incubation solution used contained the following: dl-3-glycerophosphate (disodium salt), 5 mm; 0.01% menadione; distilled water; and nitroblue tetrazolium chloride, 0.5 mg ml?1 in phosphate buffer (pH 7.4). Slides were incubated at 37��C for 20 min (Punkt et al. 1998). Controls (blanks) for both SDH and GPDH-dependent reactions were determined on serial muscle sections using solutions that omitted the relevant primary substrate to ensure that there were no non-specific GW786034 redox reactions (Reichmann et al. 1982). All chemicals were purchased from Sigma-Aldrich. Four low-powered images (��200 magnification) were obtained using an Olympus BX51 microscope and an Olympus DP71 camera in the same preset lighting conditions for all slides. Optical density (OD) was measured, using Scion Image? software (Scion Corporation, Frederick, MD, USA), as an index of muscle oxidative (SDH images) and glycolytic capacity (GPDH images). Images were converted to grey scale; each pixel was quantified as one of 256 greys. Transmission of 100% light was equivalent to a grey scale value of zero, and 0% light transmission was equal to a value of 256. Using an uncalibrated OD function, grey scale values were converted to uncalibrated OD values. All data were first averaged per animal and then Adenosine per group and are expressed as means �� SEM. Statistical comparisons between groups were performed using Student's unpaired t test, one-way ANOVA or two-way ANOVA as appropriate. The Neuman�CKeuls post hoc test was applied. A value of P