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The USDA trypsin inhibitor study. I. Background, objectives, and procedural details

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Abstract

Short-term feeding studies have shown that raw soy flour and purified trypsin inhibitors (TI) cause pancreatic hypertrophy and hyperplasia in certain monogastric animals. Prolonged exposure to high levels of TI in raw soy ultimately leads to pancreatic nodular hyperplasia and acinar cell adenoma in rats exposed to low levels of pancreatic carcinogens. Such change has been observed even in the absence of any exposure to known carcinogens. These results emphasized the need for chronic (2 yr) feeding trials which would clearly reveal dose response relationships of alterations in pancreatic pathology to dietary TI and possible interactions with dietary protein and fat.

Here we report on the objectives and designs of the first phase of the USDA TI Study, including composition of diets, preparation, and analyses of test substances. To provide the requisite TI and protein levels, raw and heated defatted soy flours, soy protein isolates, and lactic casein were used. The interrelationship of dietary level of TI (93–1270 mg/100 g diet) and that of protein (10%, 20%, and 30%) to pancreatic function and pathology will be reported in the following three papers. Both serial and chronic sacrifice regimes, respectively, were employed with 26 diets and 40 weaning male Wistar rats per dietary group.

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References

  1. American Oil Chemists' Society (1979) Procedings of World Conference on Vegetable Food Proteins. Amsterdam, Netherlands: J Am Oil Chem Soc 56:99–483

    Google Scholar 

  2. Andren-Sandberg A, Ihse I (1983) Regulatory effects on the pancreas of intraduodenal pancreatic juice and trypsin in the Syrian golden hamster. Scand J Gastroenterol 18:697–706

    PubMed  Google Scholar 

  3. Association of Official Analytical Chemists (1980) Method No. 7.009 Official Methods of Analysis (13th ed) Washington, DC, p 125

  4. Brabender Rapid Moisture Tester (1970) Instructional Manual 10-1, CW Brabender Instruments, Inc., South Hackensack NJ, pp 1–11

    Google Scholar 

  5. Chen I, Mitchell HL (1973) Trypsin inhibitors in plants. Phytochemistry 12:327–330

    Google Scholar 

  6. Churella HR, Yao BC, Thomson WHB (1976) Soybean trypsin inhibitor activity of soy infant formulas and its nutritional significance in rats. J Agric Food Chem 24:393–397

    PubMed  Google Scholar 

  7. Cohen S (1980) Pathogenesis of coffee-induced gastrointestinal symptoms. N Engl J Med 303:122–124

    PubMed  Google Scholar 

  8. Corn Industries Research Foundation (1965) Protein Method A-18. Standard Analytical Methods (1st ed) Washington, DC

  9. Crass RA, Morgan, RGH (1981) Rapid changes in pancreatic DNA, RNA and protein in the rat during pancreatic enlargement and involution. Inter J Vitam Nutr Res 51:85–91

    Google Scholar 

  10. Crass, RA, Morgan, RGH (1982) The effect of long-term feeding of soya-bean flour diets on pancreatic growth in the rat. Br J Nutr 47:119–129

    PubMed  Google Scholar 

  11. Creutzfeldt, W (1982) Gastrointestinal peptides-role in pathophysiology and disease. Scand J Gastroenterol 77:7–20

    Google Scholar 

  12. Davies, MG, Thomas, AJ (1973) An investigation of hydrolytic techniques for the amino acid analysis of foodstuffs. J Sci Food Agric 24:1525–1540

    PubMed  Google Scholar 

  13. del Rio MD, Minetti M, Ferrer PR, de Valencia ME, Sambucetti ME, Sanahuja JC (1981) The nutritive value of soy based dry-mix soups formulated with different soy products. Nutr Rep Int 24:67–73

    Google Scholar 

  14. Dlugosz J, Folsch UR, Creutzfeldt WC (1983) Inhibition of intraduodenal trypsin does not stimulate exocrine pancreatic secretion in man. Digestion 26:197–204

    PubMed  Google Scholar 

  15. Doell BH, Ebden CJ, Smith CA (1981) Trypsin inhibitor activity of conventional foods which are part of the British diet and some soya products. Qual Plant Plant Foods Hum Nutr 31:139–150

    Google Scholar 

  16. Fazio T, White RH, Dusold LR, Howard JW (1973) Nitrosopyrrolidine in cooked bacon. J Assoc Off Agric Chem 56:919–921

    Google Scholar 

  17. Fiddler RN (1977) Collaborative study of modified AOAC method for analysis of nitrites in meat and meat products. J Assoc Off Agric Chem 60:594–599

    Google Scholar 

  18. Figarella C, Negri GA, Guy O (1975) The two human trypsinogens. Eur J Biochem 53:457–463

    PubMed  Google Scholar 

  19. Folsch UR, Winckler K, Wormsley KG (1978) Influence of repeated administration of cholecystokinin and secretion on the pancreas of the rat. Scand J Gastroenterol 13:663–671

    PubMed  Google Scholar 

  20. Folsch UR, Wormsley KG (1974) The pancreatic secretion of enzymes in rats treated with soybean diet. Scand J Gastroenterol 9:679–683

    PubMed  Google Scholar 

  21. Green GM, Lyman RL (1972) Feedback regulation of pancreatic enzyme secretion as a mechanism for trypsin inhibitor-induced hypersecretion in rats. Proc Soc Exp Biol Med 140:6–12

    PubMed  Google Scholar 

  22. Green GM, Miyasaka K (1983) Rat pancreatic response to intestinal infusion of intact and hydrolyzed protein. Am J Physiol 245 (Gastrointest Liver Physiol 8): G394-G398

    PubMed  Google Scholar 

  23. Green GM, Nasset ES (1977) Importance of bile in regulation of intraluminal proteolytic enzyme activities in the rat. Gastroenterol 79:695–702

    Google Scholar 

  24. Green GM, Nasset ES (1983) Role of dietary protein in rat pancreatic enzyme secretory response to a meal. J Nutr 113:2245–2252

    PubMed  Google Scholar 

  25. Green GM, Olds BA, Matthews G, Lyman, RL (1972) Protein as a regulator of pancreatic enzyme secretion in the rat. Proc Soc Exp Biol Med 142:1162–1167

    Google Scholar 

  26. Gumbmann MR, Spangler WL, Dugan GM, Rackis JJ, Liener IE (1985) The USDA trypsin inhibitor study. IV. The chronic effects of soy flour and soy protein isolate on the pancreas in rats after two years. Qual Plant Plant Foods Hum Nutr 35:

  27. Gunn RA, Taylor PR, Gangaros EJ (1980) Gastrointestinal illness associated with consumption of a soybean protein extender. J Food Sci 43:525–527

    Google Scholar 

  28. Hamestrand GE, Black LT, Glover JD (1981) Trypsin inhibitors in soybean products. Modification of the standard analytical procedure. Cereal Chem. 58:42–45

    Google Scholar 

  29. Hasdai A, Liener IE (1983) Growth, digestibility and enzymatic activities in the pancreas and intestines of hamsters fed raw and heated soy flour. J Nutr 113:662–668

    PubMed  Google Scholar 

  30. Hotz J, Ho SB, Go VLW, DiMagno EP (1983) Short-term inhibition of duodenal tryptic activity does not affect human pancreatic bilary or gastric function. J Lab Clin Med 101:488–495

    PubMed  Google Scholar 

  31. Ihse I, Lilja P, Lindquist I (1977) Feedback regulation of pancreatic enzyme secretion by intestinal trypsin in man. Digestion 15:303–308

    PubMed  Google Scholar 

  32. Jacobson D, Tillman R, Toskes P, Curington C (1982) Acute and chronic feedback inhibition of pancreatic exocrine secretion in human subjects (abstract). Gastroentrol 82:1091

    Google Scholar 

  33. Johnson LR (1981) Effects of gastrointestinal hormones on pancreatic growth. Cancer 47:1640–1645

    PubMed  Google Scholar 

  34. Kakade ML, Rackis JJ, McGhee JE, Puski G (1974) Determination of trypsin inhibitor activity of soy products collaborative analysis of an improved procedure. Cereal Chem 51:376–382

    Google Scholar 

  35. Kakade ML, Simons N, Liener IE (1969) An evaluation of natural vs synthetic substrates for measuring the antitryptic activity of soybean samples. Cereal Chem 46:518–526

    Google Scholar 

  36. Krawisz BR, Miller LJ, DiMagno EP, Go VLW (1980) In the absence of nutrients, pancreatic bilary secretions in the jejenum do not exert feedback control of human pancreatic or gastric function. J Lab Clin Med 95:13–18

    PubMed  Google Scholar 

  37. Krogdahl A, Holm H (1979) Inhibition of human and rat pancreatic proteinases by crude and purified soybean proteinase inhibitors. J Nutr 109:551–558

    PubMed  Google Scholar 

  38. Lebenthal E, Choi TS, Lee PC (1981) The development of pancreatic function in premature infants after milk-based and soy based formulas. Pediatric Res 15:1240–1241

    Google Scholar 

  39. Levison DA, Morgan RGH, Brimacombe JS, Hopwood D, Coghill G, Wormsley KG (1979) Carcinogenic effects of di-(2-hydroxypropyl)nitrosamine (DHPN) in male Wistar rats: Promotion of pancreatic cancer by a raw soya flour diet. Scand J Gastroenterol 14:217–224

    PubMed  Google Scholar 

  40. Lewis JH, Taylor FHL (1947) Comparative utilization of raw and autoclaved soybean protein by the human. Proc Soc Exp Biol Med 64:85–87

    Google Scholar 

  41. Liener IE (1981) Factors affecting the nutritional quality of soya products. J Am Oil Chem Soc 58:406–415

    Google Scholar 

  42. Liner IE, Kakade ML (1980) Protease inhibitors. In: Liener IE (ed) Toxic constituents of plant foodstuffs. New York: Academic Press, pp 7–71

    Google Scholar 

  43. Liener IE, Nitzan Z, Srisangnam C, Rackis JJ, Gumbmann MR (1985) The USDA trypsin inhibitor study. II. Time-related changes in the pancreas of rats Qual Plant Plant Foods Hum Nutr 35:

  44. Life Sciences Research Office of Federation of American Societies for Experimental Biology (1979) Evaluation of the Health Aspects of Soy Protein Isolates as Food Ingredients Scogs — 101. Prepared for Bureau of Foods, Food and Drug Administration, Washington, DC, under contract no. FDA 223-75-2004 by Life Sciences Office, Besthesda, MD, pp4 and A-1 to A-27

  45. Mc Guinness EE, Hopwood D, Wormsley KG (1982) Further studies of the effects of raw soya flour on the rat pancreas. Scand J Gastroenterol 17:273–277

    PubMed  Google Scholar 

  46. McGuinness EE, Morgan RGH, Levison DA, Frape DL, Hopwood G, Wormsley KG (1980) The effects of long-term feeding of soya flour on the rat pancreas. Scand J Gastroenterol 15:497–502

    PubMed  Google Scholar 

  47. McGuinness EE, Morgan RGH, Levison DA, Hopwood D, Wormsley KG (1981) Interaction of azaserine and raw soya flour on the rat pancreas. Scand J Gastroenterol 16:49–56

    PubMed  Google Scholar 

  48. Miyasaka K, Green GM (1984) Effect of partial exclusion of pancreatic juice on rat basal pancreatic secretion. Gastorenterol 86:114–119

    Google Scholar 

  49. Moore S (1963) On the determination of cystine as cysteic acid. J Biol Chem 238:235–237

    Google Scholar 

  50. Morgan RGH, Levison DA, Hopwood D, Saunders JHB, Wormsley KG (1977) Potentiation of the action of azaserine on the rat pancreas by raw soya bean flour. Cancer Letts 3:87–90

    Google Scholar 

  51. Moroz LA, Young WH (1980) Kunitz soybean trypsin inhibitor. A specific allergen in food amphylaxis. New Engl J Med 302:1126–1128

    PubMed  Google Scholar 

  52. Murthy SNS, Dinoso VP, Clearfield HR, Chey WY (1977) Simultaneous measurement of basal pancreatic, gastric acid secretion, plasma gastrin, and secretin during smoking. Gastroenterol 73:758–761

    Google Scholar 

  53. Nonclinical Laboratory Studies Good Laboratory Practices (GLP) Regulations Title 21 Part 58 in Code of Federal Regulations (1978) Federal Register Vol 43 No 247 Part II December 22 pp 59986–60025

  54. Pap A, Berger Z, Varro V (1981) Trophic effect of cholecystokinin-octapeptide in man-a new way in the treatment of chronic pancreatitis? Digestion 21:163–168

    PubMed  Google Scholar 

  55. Pour PM, Rung RG, Birt D, Gengell R, Lawson T, Nagel D, Wallcave L, Salmazi SZ (1981) Current knowledge of pancreatic carcinogenesis in the hamster and its relevance to human disease. Cancer 47:1573–1587

    PubMed  Google Scholar 

  56. Rackis JJ (1974) Biological and physiological factors in soybeans. J Am Oil Chem Soc 51:161A-174A

    PubMed  Google Scholar 

  57. Rackis JJ, Gumbmann MR (1981) Protease inhibitors: Physiological properties and nutritional significance. In: Ory RL (ed) Antinutrients and Natural Toxicants in Foods. Westport CT: Food and Nutrition Press, pp 203–237

    Google Scholar 

  58. Rackis JJ, McGhee JE, Booth AN (1975) Biological threshold levels of soybean trypsin inhibitors by rat bioassay. Cereal Chem 52:85–92

    Google Scholar 

  59. Rackis JJ, McGhee JE, Gumbmann MR, Booth AN (1979) Effects of soy proteins containing trypsin inhibitors on long-term feeding studies in rats. J Am Oil Chem Soc 56:162–168

    PubMed  Google Scholar 

  60. Roebuck BD, Yager Jr JD, Longnecker DS (1981) Dietary modulation of azaserine-induced pancreatic carcinogenesis in the rat. Cancer Res 41:888–893

    PubMed  Google Scholar 

  61. Roebuck BD, Yager Jr JD, Longnecker DS, Wilpone SA (1981) Promotion by unsaturated fats of azaserine-induced pancreatic carcinogenesis in the rat. Cancer Res 41:3961–3966

    PubMed  Google Scholar 

  62. Schneeman BA, Lyman RL (1975) Factors involved in the intestinal feedback regulation of pancreatic enzyme secretion in the rat. Proc Soc Exp Biol Med 148:897–903

    PubMed  Google Scholar 

  63. Senti FR (1982) Annotated bibliographies onpancreatic changes in experimental animals fed soybeans, processed soybean products, soybean trypsin inhibitor, or cholecystokinin-pancreozymin and anti-nutritional factors in processed soybean products. Prepared for Bureau of Foods, Food and Drug Administration, Washington DC under contract no. FDA 223-79-2275 by Life Sciences Office, Federation of American Societies for Experimental Biology, Bethesda MD, pp 1–68

  64. Shibasaki M, Suzuki S, Tajima S, Nemoto N, Kuroume T (1980) Allergenicity of major component proteins of soybeans. Int Arch Allergy Appl Immunol 61:441–448

    PubMed  Google Scholar 

  65. Singh M, Webster III PD (1981) Pancreatic exocrine secretion. Clinics in Gastroenterol 10:555–581

    Google Scholar 

  66. Sissons JW (1982) Effects of soya-bean products on digestive processes in the gastrointestinal tract of preruminant calves. Proc Nutr Soc 41:53–61

    Google Scholar 

  67. Sissons JW, Nyrup A, Kilshaw PJ, Smith RH (1982) Ethanol denaturation of soya bean protein antigens. J Sci Food Agric 33:706–710

    Google Scholar 

  68. Slaff J, Toskes P, Jacobson D, Curington C (1983) Further studies of feedback inhibition of pancreatic exocrine secretion (abstract). Clin Res 31:477A

    Google Scholar 

  69. Smith AK, Circle SJ (eds) (1972) Soybeans. Chemistry and Technology Vol I Proteins. Westport CT: Avi Publishing

    Google Scholar 

  70. Spangler WL, Gumbmann MR, Liener IE, Rackis JJ (1985) The USDA trypsin inhibitor study. III. Sequential development of pancreatic pathology in rats. Qual Plant Plant Foods Hum Nutr 35:

  71. Speck ML (ed) (1976) Compendium of methods for the microbiological examination of foods. American Public Health Association. Intersociety/Agency Committee on Microbiological Methods for Foods, American Public Health Assciation, Washington DC

  72. Struthers BJ, MacDonald JR, Prescher EE, Hopkins DT (1983) Influence of several plant and animal proteins on rat pancreas. J Nutr 113:1503–1512

    PubMed  Google Scholar 

  73. Symposium: Current Progress in Pancreatic Carcinogenesis research (1975) In: Cancer Res 35:2225–2294

    Google Scholar 

  74. Symposium: Current Progress in Pancreatic Carcinogenesis research (1975) In: Cancer Res 35:2225–2294

  75. Symposium: Nutrition in the causation of cancer (1975) In: Cancer Res 35:3231–3550

    Google Scholar 

  76. Technicon Instrument Corporation (1974) Sodium nitrate and nitrite in meat extract. Method 230 and 230-72A. Technicon Instrument Corporation, Tarrytown NY

    Google Scholar 

  77. Temler RS (1980) Alterations in the pancreas of rats fed on different levels of soya flour and casein. Int J Vitam Nutr Res 50:212–213

    Google Scholar 

  78. Temler RS, Dormond CA, Simon E, Morel E (1982) The effect on rat pancreas of feeding Kunitz soya bean inhibitor or of repeated injections of purified cholecystokinin. Int J Vitam Nutr Res 52:2230

    Google Scholar 

  79. Temler RS, Dormond CA, Simon E, Morel B, Mattraux C (1984) Response of rat pancreatic proteases to dietary proteins, their hydrolysates and soybean trypsin inhibitor. J Nutr 114:270–278

    PubMed  Google Scholar 

  80. Temler RS, Simon E, Amiquet P (1983) Comparison of the interactions of soya bean protease inhibitors with rat pancreatic enzymes and human trypsin. Enzyme 30:105–114

    PubMed  Google Scholar 

  81. Theuer RC, Sarett HP (1970) Nutritional adequacy of soy isolate infant formulas in rats: Choline. J Agric Food Chem 18:913–916

    PubMed  Google Scholar 

  82. Treffot MJ, Laugier R, Bretholz A (1980) Increased gastrin release in chronic calcifying pancreatitis and in chronic alcoholism. Horm Metab Res 12:240–242

    PubMed  Google Scholar 

  83. Vitoria JC, Camarero C, Sojo A, Ruiz A, Rodriquez-Soriano J (1982) Exteropathy related to fish rice and chicken. Arch Dis Child 57:44–48

    PubMed  Google Scholar 

  84. Walker-Smith J (ed) (1979) Diseases of the small intestine in childhood. Second edition. London: Pitman Medical, pp 139–170

    Google Scholar 

  85. Warner K, Bookwalter GN, Rackis JJ, Honig DH, Hockridge E, Kwolek WF (1982) Prevention of rancidity in experimental rat diets for long-term feeding. Cereal Chem 59:175–178

    Google Scholar 

  86. Yanatori Y, Fujita T (1976) Hypertrophy and hyperplasia in the endocrine exocrine pancreas of rats fed soybean trypsin inhibitor or repeatedly injected with pancreozymin. Arch Histol Jap 39:67–78

    PubMed  Google Scholar 

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Rackis, J.J., Gumbmann, M.R. & Liener, I.E. The USDA trypsin inhibitor study. I. Background, objectives, and procedural details. Plant Food Hum Nutr 35, 213–242 (1985). https://doi.org/10.1007/BF01092196

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