Tobacco Documents Online

Cohen, S.M., Ellwein, L.B., Okamura, T., Masui, T., Johansson, S.L., Smith, R.A. Welmer, J.M., Khachab, M., Chappel, C.1., Schoenig, G.P., Emerson, 7.L. & Garland, E.M. (1991a). Comparative bladder tumor promoting activity of sodium saccharin, sodium ascorbate, related acids, and calcium salts in rats. Cancer Res. 51: 1766-1777. Cohen, S.M., Cano, M., Garland, E.M., Earl, R.A. & Carson, S.D. (1991b). Role of silicates and protein'in the proliferative effects of saccharin on the male rat urothelium. Carcinogenesis 12: 1551-1555. Eklund, S.H., Garland, E.M. & Cohen, S.M. (1992). Characterization of proteins of male rat urine which bind saccharin in vitro. Presented at 83rd American Association for Cancer Research Meeting, May 20-23, San Diego, CA. Elcock, M., & Morgan, R.W. (1993). Update on artificial sweeteners and bladder cancer. Reg. Toxicol. and Pharmacol. In press. Ellwein, L.B. & Cohen, S.M. (1990). The health risks of saccharin revisited. Crit. Rev. in Toxicol. 20: 311-326. Fujita, I., Yoshida, 0., Yuasa, Y., Rhim, J.S., Hatanaka, M. & Aaronson, S.A. (1984). Ha- nu oncogenes are activated by somatic alterations in human urinary tract tumors. Nature (Lond.) 309: 464-466. Fujita, J., Srivastava, S.K., Kraus, M.H., Rhim, J.S., Tronick, S.R. & Aaronson, S.A. (1985). Frequency of molecular alterations affecting m protoncogenes in human urinary tract tumors. Proc. Natl. Acad. Sci. USA 82: 3849-3853. Fukushima, S. & Cohen, S.M. (1980). Saccharin-induced hyperplasia of the rat urinary bladder. Cancer Res. 40: 734-736. Fukushima, S., Shirai, T., Hirose, M. & Ito, N. (1986a). Significance of L-ascorbic acid and urinary electrolytes in promotion of rat bladder carcinogenesis. jn: Diet, Nutrition and Cancer (Y. Hayashi gkg(., Eds.), pp. 159-168. Jpn. Sci. Soc. Press. Tokyo/VNU Sci. Press, Utrecht. Fukushima, S., Thamavit, W., Kutata, Y. & Ito, N. (1986b). Sodium citrate: a promoter of bladder carcinogenesis. Jpn. J. Cancer Res. (GANN) 77: 1-4. Garland, E.M., Sakata, T., Fisher, M.J., Masui, T. & Cohen, S.M. (1989). Influences of diet and strain on the proliferative effect on the rat urinary bladder induced by sodium saccharin. Cancer Res. 49: 3789-3794. Garland, E.M., St. John, M., Eklund, S.H., Mattson, B.J., Johnson, L.S., Cano, M. & Cohen, S.M. (1993). ` A comparison of the effects of sodium saccharin in NBR rats and in intact and castrated male P344 rats. Cancer Research. In press. 11

Hasegawa, R. & Cohen, S.M. (1986). The effect of different salts of saccharin on the rat urinary bladder. Cancer Lett. 30: 261-268. Hasegawa, R., So, J.M.K., Cano, M., lssenberg, P., Klein, D.A., Walker, B.A., Jones, J.W., Schnell, R.C., Merrick, B.A., Daview, M.H., McMillan, D.T. & Cohen, S.M. (1984). Bladder freeze ulceration and sodium saccharin feeding in the rat: Examination for urinary nitrosamines, mutagens and bacteria, and effects on hepatic microsomal enzymes. Fd. Chem. Toxicol. 22: 935-942. Herbold, B.A. (1981). Studies to evaluate artificial sweeteners, especially Remsen-Fahlberg saccharin, and their possible impurities, for potential mutagenicity by the Salmonella/mammalian liver microscome test. Mutat. Res. 90: 365-372. Howe, G.R., Burch, J.D., Miller, A.B., Morrison, B., Gordon, P., Weldon, L., Chambers, L.W., Fodor, G. & Winsor, G.M. (1977). Artificial sweeteners and human bladder cancer. Lancet 11: 578. IARC (1980). International Agency for Research on Cancer Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. Some Non-Nutritive Sweetening Agents. Vol. 22, p. 184. 1ARC, Lyon. IARC (1982). International Agency for Research on Cancer Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. ChemicaLc, Industrial Processes and Industries Associated with Cancer in Humans. Suppl. 4, p. 224. IARC, Lyon. JECFA (1984). Joint FAOIWHO Expert Committee on Food Additives. Toxicological Evaluation of Certain Food Additives and Food Contaminants. Twenty-eighth report of the Joint FAO/WHO Expert Committee on Food Additives. March 19-28. Geneva, WHO Technical Report Series No. 710. Lina, B.A.R. & Woutersen, R.A. (1989). Effects of urinary potassium and sodium ion concentrations and pH on N-butyl-N-(4-hydroxybutyl)nitrosamine-induced urinary bladder carcinogenesis in rats. Carcinogenesis 10: 1733-1736. Lutz, W.K. & Schlatter, Ch. (197'n. Saccharin does not bind to DNA of liver or bladder in the rat. Chem: Biol. Interactions 19: 153. Masui, T., Mann, A.M., Garland, E.M., Okamura, T., Johansson, P.L. & Cohen, S.M. (1990). Point mutations in codons 12 and 61 of the Ha-In gene in rat urinary bladder carcinomas induced by n-[4-(5-nitto-2-furyl)-2-thiazolyl] formamide. Molecular Carcinogenesis 3: 210-215. Masui, T., Mhmt, A.M., Macatee, T.L., Okamura, T., Garland, E.M., Fujii, H., Pelling, J.C. & Cohen, S.M. (1991). H-ia mutations in rat urinary bladder carcinomas induced by N-[4-(5- nitro-2-furyl)-2-thiazolyl]formamide and sodium saccharin, sodium ascorbate, or related salts. Cancer Res. 51: 3471-3475. 12 N tn 0 N s .P ~ N W N

Momose, H., Kakinuma, H., Shariff, S.Y., Mitchell, G.B., Rademaker, A. & Oyasu, R. (1991). Tumor-promoting effect of urinary epidermal growth factor in rat urinary bladder carcinogenesis. Cancer Res 51: 5487-5490. Morgan, R.W. & Wong (1985). A review of epidemiological studies on artificial sweeteners and bladder cancer. Fd. Chem. Toxicol. 23, 529-533. Nakanishi, K., Hagiwara, A., Shibata, M., Imaida, K., Tatematsu, M. & Ito, N. (1980). Dose response of saccharin in induction of urinary bladder hyperplasias in Fischer 344 rats pretreated with n-butyl-n-(4-hydroxybutyl)nitrosanrine. JNCI 65: 1005-1010. Nakanishi, K., Fukushima, S., Hagiwara, A., Tamano, S. & Ito, N. (1982). Organ-specific promoting effects of phenobarbital sodium and sodium saccharin in the induction of liver and urinary bladder tumors in male F344 rats. J. Natl. Cancer Inst. 68: 497-500. Neal, D.E., Bennett, M.K., Hall, R.R., Marsh, C., Abel, P.D., Sainsbury, J.R.C. & Harris, A.L. (1985). Epidermal-growth-factor receptors in human bladder cancer: comparisons of invasive and superficial tumours. Lancet, Feb. 16. Renwick, A.G. (1993). A data-derived safety (uncertainty) factor for the intense sweetener, saccharin. To be published. Riggin, R.M., Margard, W.L. & Kinzer, G.W. (1983). Characterization of impurities in commercial lots of sodium saccharin produced by the Sherwin-Williams process. II. Mutagenicity. Fd. Chem. Toxicol. 21: 11-17. Risch, H.A., Burch, J.D., Miller, A.B., Hill, G.B., Steele, R. & Howe, G.R. (1988). Dietary factors and the incidence of cancer of the urinary bladder. Am. J. Epidemiol. 127: 1179-1191. Schoenig, G.P. & Anderson, R.L. (1985). The effects of high dietary levels of sodium saccharin on mineral and water balance and related parameters in rats. Fd. Chem. Toxicol. 23, 465-474. Schoenig, G.P., Goldenthal, E.L., Geil, R.G., Firth, C.H., Richter, W.R. and Carlborg, F.W. (1985). Evaluation of the dose response and in utero exposure to saccharin in the rat. Fd. Chem. Toxicol. 23: 475-490. Shibata, M.-A., Kagawa, M., Kawabe, M., Hagiwara, A. & Fukushima, S. (1991). Comparative promoting activities of phosphate salts on rat two-stage bladder carcinogenesis under conditions of equivalent urinary Na* or K+ levels. Teratogenesis, Carcinogenesis, Mutagenesis. 11: 305-313. Squire, R.A..(1985). Histopathological evaluation of rat urinary bladders from the IRDC two- generation bioassay of sodium saccharin. Fd. Chem. Toxicol. 23: 491-497. Sweatman, T.W. & Renwick, A.G. (1979). Saccharin metabolism and tumorigenicity. Science 205: 1019-1020. N N O N ~ ~ o> N w w 13

Sweatman, T.W. & Renwick, A.G. (1982). Tissue levels of saccharin in the rat during two- generation feeding studies. Toxicol. Appl. Pharmacol. 62, 465-473. Taylor, J.M., Weinberger, M.A. & Friedman, L. (1980). Chronic toxicity and carcinogenicity to the urinary bladder of sodium saccharin in the in uto4-exposed rat. Toxicol. Appl. Pharmacol. 54: 57-75. Tisdel, M.O., Nees, P.O., Harris, D.L. & Derse, P.H. (1974). Long-term feeding of saccharin in rats. In: Symposium: Sweeteners. (G.E. Inglett, Ed.), pp. 145-158. The AVI Publishing Company, Inc., Westport, Conn. Verma, A.K. Bryan, G.T. & Reznikoff, C.A. (1985). Tumor promoter 12-0-tetradecanoyl- phorbol-13-acetate receptors in normal human transitional epithelial cells. Carcinogenesis 6: 427-432. Weisburger, J.H. (1988). Carcinogenesis bioassays of natural and artificial sweeteners. In: Sweeteners: Health Effects. pp. 193, 224. Princeton Scientific Publishing Company, Princeton. Williamson, D.S. & Cohen, S.M. (1989). Unpublished observations. 14

Table 1. Bladder Tumour Incidence in F1-Genaation Male Rats in Two-Generation Bioassays of NaS . FDA Study' WARF Study' CHPB Study' IRDC Study' Dietary Level # per % Group Incidence # per % Group Incidence # per % Group Incidence # per % Group Incidence 0 29 3.0 20 0 42 0 324 0" 0.01 16 0 - - - - - - 0.05 - - 20 0 - - 0.1 27 0 - - - - 0.5 - - 20 5.0 - - 1.0 22 0 - - - - 658 0.8 3.0 - - - - - - 472 1.7* 4.0 - - - - - - 189 6.3* 5.0 21 5.0 20 35* 45 26.9* 120 12.5* 6.25 - - - - - - 120 16.7* 7.5 23 30.0* - - - - 118 31.4* ' Food and Drug Administration, reported by Taylor ". (1980); Wisconsin Alumni Research Foundation, reported by Tisdel gl,g(. (1974); Canadian Health Protection Branch, reported by Arnold j„h(. (1980a); International Research and Development Corp., reported by Schoenig ". (1985). ` The background incidence for total bladder neoplasia at IRDC is 0.8 % * Statistically significant SEZ9bLZOSZ

Bladder Hyperplasia (46) 30 20 a 10 a 0 0 • Labelling Index 0.8 0.6 ~ 0.4 ~ 0 0.0 A + • 2 4 6 Dietary NaS (96) C + • T 2 4 6 Dietary NaS (%) + • + • 8 8 Bladder Tumors (gh) 40 0 30 - 20 a t0 a Light Transmittance (%) 100 Dietary NaS (9'0) + 0 2 4 T 6 Dietary NaS (qo) 1 • • 80 - 40 - 20 - B D 2 4 T 6 + 8

Dietary NaS 'I Prevented by Increased urine pH and sodium concentration NH.CI in the diet -~-4 -• or AIN 76A diet Increased urine volume and decreased osmolality a High urinary protein levels in male vs female (au-globulin) • Absent or less -~~-• Urinary amorphous precipitate and crystals in female rats I Urothelial cytotoxicity ~ Absent or less r+ Urothelial hyperplasia and proliferation in female rats Bladder tumors Bladder tumor promotion in long-term bioassays in initiated rats N . N O IV ~ A O N W V

Legend for Figures Figu~ re 1. Incidence of Urothelial Hyperplasia (A) and Bladder Tumors (B) in the IRDC Study of NaS in the Rat (Squire, 1985). Labelling Index Data (C) Taken from Cohen (1990), and Light Transmittance Findings (D) from a Recent Study (Cohen, Unpublished). Figure 2. Sequence of Urinary and Bladder Changes Observed in Male Rats Fed High Dietary Levels of NaS IV N O . N i 44, a) N W ~ f t