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Quality control of histamine and methacholine in diagnostic solutions with capillary electrophoresis.

Solutions of histamine and methacholine bromide in different matrices for diagnostic purposes were analyzed for stability and quality control using capillary electrophoresis. Histamine (2,[4-imidazolyl]ethylamine) [CAS No. 51-45-6] was determined using a 0.1 M Tris-borate buffer of pH 8.3 with 5.10(-5) M cetyltrimethylammonium bromide (CTAB) and 0.005% poly(vinyl alcohol) (PVA) and detected at 214 nm using clenbuterol [4-amino-alpha-(tert.-butylaminomethyl)-3,5-dichlorobenzyl alcohol] [37148-27-9] as an internal standard. Metacholine bromide (acetyl-beta-methacholine bromide) [333-31-3] was determined with a 0.01 M creatinine-chloride buffer of pH 4.85 and detected with indirect UV at 230 nm using potassium as an internal standard. Histamine solutions were stable for a prolonged period of time, whereas under enforced degradation conditions methacholine was hydrolyzed, yielding acetic acid and (tentatively) beta-methylcholine as reaction products.

Drug Stability↗

Relaxation decreases large-airway but not small-airway asthma.

Eleven asthmatic subjects were each offered sixteen sessions of relaxation therapy, consisting of progressive relaxation, desensitization, and EMG biofeedback to the trapezius and frontalis areas, while 9 subjects were offered a complex placebo. Subjects in the relaxation condition showed greater improvement than subjects in the placebo condition in performance on a methacholine challenge test which, as a measure of airway reactivity, reflects degree of asthma. Subjects in both conditions showed improvements in various self-report measures of asthma symptoms and psychopathology. Subject in the relaxation condition reported a significantly greater decrease in frequency of emotional precursors to asthma attacks than subjects in the control condition. Overall improvement on the methacholine challenge test was predicted almost perfectly by heliox spirometry. Only subjects showing predominant large-airway obstruction obstruction improved on the methacholine challenge test. The relative contribution of large airway obstruction to asthma was found to correlate with psychopathology.

Adult↗

Serotonin release is mediated by muscarinic receptors on duodenal mucosal cells.

Serotonin (5-HT), found in abundance in intestinal enterochromaffin cells, has been shown to be released in response to neural, humoral, and intraluminal stimuli but the mechanisms controlling release are poorly understood. The purpose of this study was to determine whether 5-HT release induced by the muscarinic agonist, methacholine, is mediated by enteric nerves or is a direct action at the mucosal cell level. We mounted rabbit mucosal sheets containing intact submucosal neural elements, but stripped of muscularis propria and myenteric plexus, in modified Ussing chambers and measured the release of 5-HT in response to 5 X 10(-5) M methacholine added to both mucosal and serosal surface bathing solutions, in the presence and absence of selective neural blockade with 1 X 10(-6) M tetrodotoxin. 5-HT release in control tissues (no drugs) as measured by radioimmunoassay was 25.7 +/- 6 ng/cm2/30 min (mean +/- SEM). In the presence of mucosal and serosal methacholine, total 5-HT release increased significantly (P less than 0.05) to 66.7 +/- 9 ng/cm2/30 min. When tetrodotoxin alone was applied, 5-HT release significantly increased to 55.2 +/- 9 ng/cm2/30 min compared to matched control. In the presence of tetrodotoxin, methacholine increased 5-HT release to 79.3 +/- 9 ng/cm2/30 min, which was significantly greater than with tetrodotoxin alone. These results provide evidence of an inhibitory neural regulation of basal 5-HT release since release increased in the presence of neural blockade. They also suggest that a muscarinic receptor at or near the enterochromaffin cells mediates mucosal 5-HT release since 5-HT is further increased by methacholine even in the presence of neural blockade.

Animals↗