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To define the role of gastrin, if any, in the response of the lower esophageal sphincter (LES) to bethanechol, serum gastrin determinations and LES pressure measurements were made in controls, patients with vagotomy and antrectomy (V&A), and patients with vagotomy and pyloroplasty (V&P). Despite significant differences in mean basal serum gastrin levels no differences were found in mean resting LES pressures among these groups. In controls significant increases in LES pressure occurred after subcutaneous bethanechol, but serum gastrin levels did not change from basal values. Subcutaneous injections of bethanechol produced significantly greater increases in LES pressures in V&P patients than in V&A patients. Serum gastrin levels did not change in either group; however, background serum gastrin concentrations were significantly greater for V&P patients than V&A patients throughout the study. Intravenous infusion of human gastrin I heptadecapeptide in controls significantly increased sphincter responses to bethanechol. Thus, these studies provide evidence to suggest that the LES response to bethanechol is affected by background serum gastrin levels.