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3 Table 1. Laboratory Findings Table 2. Amino Acid Composition of Glomerular Basement Membrane (residues/1, 000 residues)

4 Table 3. Disaccharide and Heteropolysaccharide of Glomerular Basement Membrane (residues/1, 000 residues of amino acid)

5 Table 4. Effects of Immunosuppressants on Glomerular Basement Membrane of Nephrotoxic Nephritic Rat

6 S.: Automatic Recording Apparatas for Use in the Chromatography of Amino Acids. Anal. Chem. 30: , ) Isemura, M., Ikenaka, T., and Matsushima, Y.: Comparative Study of Carbohydrate-Protein Complexes. I. The Structures of Glycopeptides Derived from Cuttlefish Skin Collagen. J. Bio- chem. 74: 11-21, ) Kimura, S.: Determination of the Glycosylated Hydroxylysines in Several Invertebrate Col- lagens. J. Biochem. 71: , ) Butler, W. T., and Cunningham, L. W.: Evidence for the Linkage of a Disaccharide to Hydroxylysine in Tropocollagen. J. Biol. Chem. 241: , ) Halme, J., Kivirikko, K. I., and Simonos, Isolation and partial characterization of highly purified protocollagen proline hydroxylase. Biochim. Biophys. Acta 198: , ) Heymann, W., and Lund, H. Z. ; Nephrotic Syndrome in Rats. Pediatrics 7 : , ) Krakower, C. A. and Greenspon, S. A.: Localization of the Nephrotoxic Antigen within the Isolated Renal Glomerulas. 51: , A. M. A. Arch. Path. 4) Westberg, N. G., and Michael, F. N.: Human Glomerular Basement Membrane, Preparation and Composition. Biochemistry 9: , ) Moore, S., Spackman, D. H., and Stein, W. H.: Chromatography of Amino Acids on Sulfonated Polystyrene Resins. An Improved Resins. Anal. Chem., 30: , ) Spackman, D. H., Stein, W. H., and Moore, 12) Kivirikko, K. I., Shudo, K., Sakakibara, S., and Prockocp, D. J. ; Studies on Protocollagen. Lysine Hydroxylase. Hydroxylation of Synthetic Peptides and Stoichiometric Decarboxylation of a-ketoglutarate. Biochemistry, 11: , ) Grant, M. E., and Prockocp, D. J.: The Biosynthesis of Collagen (Third of Three Parts). New Engl. J. Med., 286 : , ) Piez, K. A.: Cross-linking of collagen and ela stin. Annual Rev. Biochem. edited by Boyer P. D. Annual Reviews, Inc. California, U.S. A., Vol. 37, p ) Bailey, A. J. and Peach, C. M.: Isolation andd structural identification of a labile intermolecular crosslink in collagen. Biochem. Biophys. Res. Commun., 33: , ) Kefalides, N. A., and Forsell-Knott, L.: Structural changes in the protein and carbohydrate composition of glomerular basement membrane in aminonucleoside nephrosis. Biochem. Biophys. Acta 203: 62-66, 1970.

7 17) Spiro, R. G.: Studies on the renal glomerular basement membrane. Nature of the carbohydrate units and their attachement to the peptide portion. J. Biol. Chem., 242: , ) Misra, R. P., and Berman, L. B.: Studies on Glomerular Basement Membrane V. Effects of Steroid on Membrane Chemistry and Its Protein Permeability. Lab. Invest., 26: , ) McIntosh, R. M., Kihara, Ii., Kaufman, D. B. and Kulvinskas, C.: Effects of Cyclophsophamide on the Chemical Composition of the Glomerular Basement Membrane and Non-Collagenous Urinary Glycoprotein of the Rat. Expeam 28: , Biochemical Studies on the Glomerular Basement Membrane (II) Effects of Immunosuppressants on the Glomerular Basement Membrane of Nephrotoxic Nephritic Rat. (1) Takashi Miyakawa, Yukio Nozawa, Yasutami Kinoshita, Mamoru Isemura, Takehiko Koide Second Department of Internal Medicines, (Director: Prof. Irasutami Kinoshita; Second Department of Biochemistry" (Director: Prof. Toluzi Ihena1a) Niigata University School of Medicine, Asahimachi don 1, iyiigaia City, Japan In order to investigate the action mechanisms of immunosuppreceante and c:,tablish the method of drug administration with a minimum of side-effects in the renal diseases, the effects of steroid and cyclophosphamide on the chemical composition of the renal glomerular basement membrane of normal and nephrotoxic nephritic rats were examined. The glomerular basement membrane was isolated, and amino acid, carbohydrate and hydroxylysinedisaccharide analyses were made. Nephrotic syndrome and glomerulitis were observed in the nephrotoxin. nephritis (NT) group. Slight glomerulitis was seen in the NT group given stcroi _d (NTST group) and in the NT group given cyclophosphamide (NTCY group). A slight decrease in hydroxylation of praline, decreases both in hydroxylation of lysine and in cross linking were presumed in the NT group. In the group given steroid (ST group) decreases in hydroxylation of proline and lysine and increase in cross-linking were presumed. In the group given cyclophosphamide (CY group), remarkable decrease in hydroxylation of proline was presumed. In the NTST group, increase in crosslinking was predicted, and in the NTCY group, increase in hydroxylation of lysine was presumed, as compared with the NT group. The change in the degree of hydroxylation of lysine was the most important index reflecting the effects of nephrotoxin nephritis and immunosuppressants on the glomerular basement membrane. The amount of hydroxylysine-linked disaccharide chain was paralleled with ~his change. As judged from the effects on the degree of hydroxylation of lysine and from the renal histological findings, cyclophosphamide is suggested to be more suitable than steroid for the treatment of the ncphrotoxin nephritis.

8 腎 糸 球体 幕 底 膜 の 生化 学 的 研 究(第1報) 60 Fig. 1 Electron fixed Fig. 2 in micrograph in osmium Glomerulitis the nephrotoxin of isolated glomerular basement membrane (, 11, 000; tetroxide. with plasma nephritis imbibition NT) group. (60) in moderatc-dcgree(h. E. stain cbserved

9 日 腎誌 第XVI巻 Fig. 3 Glomerulitis with in the NT group Fig. 4 Glomerulitis in slight 61 宮 川他4名 第1号1974 degree in plasma given the than imbibition steroid NT,NTST group in the in slight given NTST degree t H. E. stain's observed NTCY group observed group). cyclophosphamide, group. (61)

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