Therefore, patients and control subjects were finally selected for the study.In the alcoholic group, patients were smokers and of them heavy smokers while control subjects were smokers and of them were heavy smokers. After overnight fasting, all patients underwent baseline blood chemical evaluation as described below.During upper gastrointestinal endoscopy, biopsy samples of adequate size were taken each from the antrum and the gastric body.These serve the purpose of assessing routine histology, <a href="http://www.targetmol.com/compound/Propidium-Iodide"></a>
malonyldialdehyde level, vitamin E and glutathione concentration and for assay vitamin B binding by intrinsic factor. Further, gastric juice samples were collected and stored for assaying intrinsic factor and xanthine oxidase concentration.During such period patients continued in their dietary habits and, in particular, with unchanged alcohol consumption, as ascertained later on.The final powder was resuspended in L of chromatographic solvent A and added to a high performance liquid chromat ography system. Xanthine oxidase activity was assessed by applying to tissue the methodology cited below for gastric juice.For alpha tocopherol determination preweighed biopsies were removed from the freezer and kept in ice to then he quickly blotted and weighed.Each, biopsy sample was placed in a mL tube with traces of butylated hydroxy toluene crystals and L of a freshly made collage nase solution. On removal from the bath, L of a freshly made pronase E solution was added to each tube which was placed in ice.Each softened biopsy mixed with butylated hydroxytoluene crystals and enzymes was transferred to a glass microhomogenizer for s.The homogenate was then returned to its original tube.The tubes were finally incubated in a water bath at C lor min.Soon afterwards, a quick extraction of atocopherol was carried out as follows.Tubes were then placed in a multi vortexer for min, centrifugal at xg for min at C and the upper hexane layer was transferred to a clean mL tube.mL autosampler vials which were stored at C: the following day the specimens were injected by using the same mobile phase.To assay glutathione concentration, tissue samples were homogenized in EDTA. Excitation and emission wavelength were nm and nm, respectively.As derivati zation standards, nmo of glutathione were injected into HPLC system prior to each sample processing.The standard curvew asprep aredbyplottingincreasing concentrations of IF against B and F the radioactivity in the charcoal.IF in gastric juice was assayed using L of gastric juice treated as above and the concentrations determined from the standard curve.For xanthine oxidase determination, gastric juice samples were checked for pH value by a digital pH meter and centrifuged at rpm for min to ob ta in aclearsupe rna tant.Supe rna tants we re extracted by using a: vv chloroformethanol mixture and then centrifuged at rpm for min.Gastric juice pH was checked by a pH meter as well.This was assayed following incubation of concentrated. The specificity of the gastric homogenate IF for B was ascertained by incubating antiIF antibodies stored from the serum of patients with pernicious anemia and using normal serum as control.Either malonyldialdehyde and xanthine oxidase concentration in the gastric mucosa in alcoholics were significantly higher than in alcoholics.