No abstract

No abstract

No abstract

Although salicylate was introduced eighty years ago in the treatment of rheumatic fever (1), the mechanism of its effect is still unknown. Speculation concerning the mode of action of salicylate and its congeners has run from considerations of one or another of the physico-chemical properties of phenolic acids to a supposed effect on the pituitary-adrenal axis (2-4). In a recent study of tissue interactions of certain salicylic acid derivatives, Helander attempted to demonstrate selective localization of sodium salicylate in connective tissue using fluorescence microscopy but was unable to do so (5). From his investigations he concluded that attachment of both a carboxyl-and an amino-group to the aromatic ring is necessary for connective tissue binding. These investigations did not include gentisic acid, a diphenol which takes on special importance because in the human subject as well as in animals an appreciable portion of administered salicylate (5 to 8 per cent) is oxidized and excreted as gentisate (6)(7)(8).Numerous workers (9-12) have now confirmed the observation of Meyer and Ragan in 1948 (8) that gentisate has an antirheumatic effect entirely comparable to that of salicylate itself. The present report concerns the affinity of gentisic acid and its oxidation products for connective tissue, an experimental approach to the hypothesis that oxidation of the antirheumatic phenols is essential to their action. METHODSObservations on distribution of gentisic acid using its fluorescence in ultra-violet light. Sodium gentisate was prepared in distilled water in concentrations of 0.5 to 5.0 per cent. Mice were injected with 1.0 ml. intra-1 This study was supported in part by a grant from the U. S. Public Health Service (C-2473), and by a gift from the New York Chapter of the Arthritis and Rheumatism Foundation.2 Clinical Fellow of the American Cancer Society.Present address: Dept. of Medicine, Massachusetts General Hospital, Boston, Mass.venously and sacrificed at intervals. Dissection and observation were carried out in the near ultra-violet light (Hanovia Inspectolite, wave-length maximum 3,660 A). Equilibrium dialysis. The method used by Taggart (13) was employed, with the following modifications: Dialysis was carried out at 00 C., and tubes were inverted frequently rather than shaken constantly. Duplicate determinations were made, gentisic acid being added in one instance to the inside of the cellophane bag and in the second to the buffer on the outside. Equilibrium was attained in 24 hours. Buffer was 0.067 M sodium phosphate, pH 7.0 and 7.5, in 0.05 M NaCl, except when procollagen was tested. In this case acetate buffer pH 4.0 was used. Gentisic acid was determined by the method of Roseman and Dorfman (14) Oxidation in air of gentisate with added tissues and particles. Solutions of sodium gentisate in phosphate buffer 0.05 M, pH 7.5, were prepared in concentrations ranging from 20 to 0.04 mg. per cent. To 15 ml. of the solutions were added in separate experiments pieces of 1) dried collagen, about 150 mg.;...