MATERIALS AND METHODS Male Wistar rats were decapitated, and their brains were quickly removed and homogenized in 10 volumes of an ice-cold 0.32 M sucrose solution by using a Potter-Elvejhem glass homogenizer fitted with a Teflon pestle. The homogenate was centrifuged at 1, 000 x g for 10 min, the pellet was discarded, and the supernatant was centrifuged at 30, 000 x g for 15 min. The resultant crude mitochondrial pellet was suspended in 50 volumes of 50 mM Tris hydrochloride buffer pH 7.4 ; by dispersion with a Polytron Kinematica, Lucerne, Switzerland ; . This procedure was repeated three times, and the final suspension was frozen for at least 24 h at -800C. [3Hlmuscimol binding to GABA receptor. [3HJmuscimol was bound to GABA receptor by a slight modification of a previously described method 3 ; . Frozen membranes were thawed and pelleted 30, 000 x g, 15 min ; . The pellet was suspended in 50 volumes of 50 mM Tris hydrochloride buffer pH 7.4 ; and repelleted three times to remove endogenous GABA; it was then suspended in buffer for use in the binding assay. The standard binding assay preparation contained 300 pd of membrane material, 100 PI of [3H]muscimol 4 nM; specific activity, 19.2 Ci mmol; New England Nuclear Corp., Boston, Mass. ; , and 100 pl of buffer or test drug. After 30 min of incubation at 0 to the samples were quickly diluted by adding 5 ml of ice-cold buffer and filtered through Whatman GF B glass fiber filters Whatman, Inc., Clifton, N.J. ; . Filters were washed three times with 5 ml of ice-cold buffer, placed in scintillation vials containing 5 ml of hydroluma Lumac, The Netherlands ; , and counted in a Packard 300C liquid scintillation counter Hewlett-Packard Co., Palo Alto, Calif. ; . Specific binding was defined as the difference between the levels of binding observed in the presence or the absence of 10 p, M GABA. Receptor binding assays. [3H]quinuclidinyl benzilate binding to the muscarinic-cholinergic receptor, [3H]dihydro and alphagan. For at least 35% of the 24-hour period; this level was achieved in 90% of subjects receiving ACIPHEX 20 mg and in 100% of subjects receiving ACIPHEX 40 mg. With ACIPHEX 20 mg and 40 mg per day, significant effects on gastric and esophageal pH were noted after one day of treatment, and more pronounced after seven days of treatment. Effects on Serum Gastrin In patients given daily doses of ACIPHEX for up to eight weeks to treat ulcerative or erosive esophagitis and in patients treated for up to 52 weeks to prevent recurrence of disease the median fasting gastrin level increased in a dose-related manner. The group median values stayed within the normal range. In a group of subjects treated daily with ACIPHEX 20 mg for 4 weeks a doubling of mean serum gastrin concentrations were observed. Approximately 35% of these treated subjects developed serum gastrin concentrations above the upper limit of normal. In a study of CYP2C19 genotyped subjects in Japan, poor metabolizers developed statistically significantly higher serum gastrin concentrations than extensive metabolizers. Effects on Enterochromaffin-like ECL ; Cells Increased serum gastrin secondary to antisecretory agents stimulates proliferation of gastric ECL cells which, over time, may result in ECL cell hyperplasia in rats and mice and gastric carcinoids in rats, especially in females see Carcinogenesis, Mutagenesis, Impairment of Fertility ; . In over 400 patients treated with ACIPHEX 10 or 20 mg day ; for up to one year, the incidence of ECL cell hyperplasia increased with time and dose, which is consistent with the pharmacological action of the proton-pump inhibitor. No patient developed the adenomatoid, dysplastic or neoplastic changes of ECL cells in the gastric mucosa. No patient developed the carcinoid tumors observed in rats. Endocrine Effects Studies in humans for up to one year have not revealed clinically significant effects on the endocrine system. In healthy male volunteers treated with ACIPHEX for 13 days, no clinically relevant changes have been detected in the following endocrine parameters examined: 17 -estradiol, thyroid stimulating hormone, tri-iodothyronine, thyroxine, thyroxinebinding protein, parathyroid hormone, insulin, glucagon, renin, aldosterone, follicle-stimulating hormone, luteotrophic hormone, prolactin, somatotrophic hormone, dehydroepiandrosterone, cortisol-binding globulin, and urinary 6hydroxycortisol, serum testosterone and circadian cortisol profile. Other Effects In humans treated with ACIPHEX for up to one year, no systemic effects have been observed on the central nervous, lymphoid, hematopoietic, renal, hepatic, cardiovascular, or respiratory systems. No data are available on long-term treatment with ACIPHEX and ocular effects. Microbiology Rabeprazole sodium, amoxicillin and clarithromycin as a three drug regimen has been shown to be active against most strains of Helicobacter pylori in vitro and in clinical infections as described in the CLINICAL STUDIES and INDICATIONS AND USAGE sections. Helicobacter pylori Susceptibility testing of H. pylori isolates was performed for amoxicillin and clarithromycin using agar dilution methodology1, and minimum inhibitory concentrations MICs ; were determined. The clarithromycin and amoxicillin MIC values should be interpreted according to the following criteria: Interpretation Clarithromycin MIC g mL ; a Susceptible S ; 0.25 Intermediate I ; 0.5 Resistant R ; 1.0 a, b Interpretation Amoxicillin MIC g mL ; Susceptible S ; 0.25 a These are breakpoints for the agar dilution methodology and they should not be used to interpret results using alternative methods. b There were not enough organisms with MICs 0.25 g mL to determine a resistance breakpoint.

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