All samples and standards were assayed in duplicate. H. pylori IgG and mutant p53 were quantified by extrapolating the average optic density for each set of duplicates on a standard curve obtained with known concentrations of purified H. pylori antibodies and mutant p53

respectively. For all analyses we used a Labinstruments SLT-400 ELISA spectrophotometer (Salzburg, Austria) with a 405 nm Vorinostat ic50 filter for H. pylori and a 450 nm filter for p53 [24]. Serum ceruloplasmin was measured by nephelometry with a Behring Nephelometer buy Brigatinib 100 analyzer (Behringwerke AG, Marburg, Germany). Statistical analysis All statistical computations were performed using SPSS software package (SPSS Version 10.0 for Windows, Inc, Chicago, IL) [37]. Descriptive statistics were calculated for each variable (means and confidence click here intervals). The statistical significance of the differences between groups were analyzed by Student’s t-test or Mann-Whitney U-test. Significance of the difference between the seropositive and seronegative populations in towns with high and low mortality due to stomach cancer was found for serum concentration of p53 protein. The possible

correlations between serum ceruloplasmin concentration, H. pylori IgG antibody level and p53 level. All tests of significance were 2-tailed, and a P value of 0.05 or less were considered statistically significant. Results Helicobacter H. pylori IgG antibody (Table 1) In the coastal town of Barbate, 92 of the 308 subjects (29.87%) were positive for H. pylori IgG antibody, with a mean value of 242.5 IU/L (95% CI 232-386). Mean value

in negative subjects (n = 216) was 19.4 IU/L (CI 16-24). In the inland town of Ubrique, 257 of the 319 subjects were positive (80.56%), with a mean value of 397.3 IU/L (95% CI 345-405 IU/L). The mean value in negative subjects (n = 62) was 16.6 IU/L (CI 12-22). The difference in the rate of seropositivity in the two populations was significant at p < 0.001. Table 1 Serum concentration of anti-H. pylori IgG antibodies. Population N Mean (IU/L) CI 4-Aminobutyrate aminotransferase 95% p value BARBATE 308 ——-     H. pylori (+) 92 242.5 232-386 <0.001 H. pylori (-) 216 19.4 16-24   UBRIQUE 319 ——-     H. pylori (+) 257 397.3 345-405 <0.001 H. pylori (-) 62 16.6 12-22   GASTRIC CANCER 71 ——-     H. pylori (+) 68 400 305-495 <0.001 H. pylori (-) 3 17.4 15-19   CI, confidence interval Mutant p53 genotype (Table 2) Of the 349 subjects who were seropositive for H. pylori IgG antibody, 286 (81.94%) had mutant p53, with a mean value of 0.973 ng/mL (95% CI 0.847-1.098). Of the 278 seronegative subjects, mutant p53 protein was detected in only 27 (9.71%), with a mean value of 0.239 ng/mL (95% CI 0.131-0.346). The frequency of quantifiable mutations was thus significantly higher in subjects who were seropositive for H. pylori IgG antibody than in seronegative subjects (p < 0.001). The mean serum value was significantly higher in patients with gastric cancer (1.973 ng/mL, 95%, CI 0.895-2.