The USDA trypsin inhibitor study. IV. The chronic effects of soy flour and soy protein isolate on the pancreas in rats after two years

In two year feeding trials, histologic changes in the pancreas of the Wistar rat were evaluated after chronic dietary exposure to raw and heated, dehulled, defatted soy flour and soy protein isolates which provided a range of trypsin inhibitor (TI) concentrations from 93 to 1271 mg/100 g diet. Also investigated was the nutritional interaction of level of dietary protein with the development of pancreatic pathology. Graded levels of TI were achieved from mixtures of raw and heated soy flour or protein isolate. Dietary protein levels were 10%, 20%, and 30%; the two higher levels obtained in some diets through casein supplementation. A total of 26 diets, including casein controls, were fed to groups of 40 male rats. Growth rates with these diets were commensurate with protein quality and level. Mortality rates tended to be slightly greater in the higher protein diets, and rats fed only raw soy as a source of protein survived well.Two lesions, nodular hyperplasia (NH) and acinar adenoma (AA) constituted the major pathological findings in the pancreas, and the incidence rates of both were positively associated with the concentration of dietary TI. The incidences of pancreatic NH and AA compared to those of the corresponding casein control were significantly elevated in rats fed heated soy flour supplying 10% protein and the lowest level of TI investigated (93 mg/100 g diet). At low levels of TI (215 mg/100 g diet), supplementation by casein to raise the concentration of protein from 10% to 20% and 30% reduced the frequency of NH and AA. When the dose-response data for NH and AA was linearized by plotting the probit of percent incidence against log of TI concentration per gram of dietary protein, the response to soy flour and soy protein isolate was not significantly different. Numerous lesions commonly found in the aging rat were diagnosed, and modulation of their rate of occurrence by nutritional factors associated with the diets was apparent in some instances.Reference to a company and/or product named by the Department is only for purposes of information and does not imply approval or recommendation of the product to the exclusion of others which may also be suitable.     

The USDA trypsin inhibitor study. II. Timed related biochemical changes in the pancreas of rats

A total of 600 rats was distributed equally among 15 diets formulated from mixtures of raw and toasted soy flour and casein so as to provide 5 different levels of trypsin inhibitor (TI) at 3 levels of protein (10%, 20%, and 30%). Animals were sacrificed at the end of 6 months and at 3-month intervals thereafter over a period of 22 months. Increases in the secretory activity of the pancreas, as measured by trypsin and chymotrypsin activities in the pancreas and small intestines, were associated with increasingly higher levels of TI and protein but did not change appreciably with time. Hypertrophy and hyperplasia of the pancreas, as reflected by changes in pancreas weight and RNA and DNA content, developed at an early stage (6 months) and were likewise positively correlated with the levels of TI and protein in the diet. Although the hypertrophic response remained unchanged with time, hyperplasia became more pronounced as the period of exposure to TI was prolonged. The peptide test confirmed the stimulatory effect of TI on the pancreas, but some functional impairment was noted in those animals which had been fed raw soy flour for 18 months of or longer. The appearance of nodules on the pancreas was observed by gross inspection after 15 months, and these became larger and more numerous with time. The incidence of nodules was highly correlated with the level of TI in the diet and was highest when the diet contained 20% protein.     

The USDA trypsin inhibitor study. III. Sequential development of pancreatic pathology in rats

Weanling male, Wistar rats (40 per diet) were maintained on 15 diets which provided 4 graded trypsin inhibitor (TI) levels (93, 215, 337, and 577 mg/100g diet) repeated at 3 levels of protein (10%, 20%, and 30%). Raw and heated (toasted) soy flour provided 10% protein in the diet, while casein was added to increase the protein level to 20% and 30%. Three diets containing only casein as the protein source, at 10%, 20%, and 30% were included. Histologic changes occurring in the pancreas were evaluated in approximately 5 rats from each diet at 3 month intervals, beginning at 6 months, for 22 months. Criteria for the diagnosis of the observed histologic changes in the pancreas are discussed. Ultrastructure of the endocrine cells in the gastrointestinal mucosa was also examined.Pancreatic acinar cell density, as determined morphometrically, was reduced throughout the study in rats fed the 577 mg TI/100 g diet, indicating hypertrophy at the cellular level. Nodular hyperplasia was observed in the first sacrifice group at 6 months. Incidence of the lesion was positively related to both time of exposure and level of dietary TI. Acinar adenoma was first observed at 18 months and was most prevalent in rats fed the highest concentration of TI.Endocrine cells of the duodenal mucosa which contain cholecystokinin (so-called I cells) contained a denser population of secretory granules in animals fed the high TI diet compared with I cells from control animals, leading to the speculation that production and secretion of cytoplasmic hormones was increased in TI treated rats.Reference to a company and/or product named by the Department is only for purposes of information and does not imply approval or recommendation of the product to the exclusion of others which may also be suitable.     

Effects of raw soy flour feeding in weanling pigs: comparison with rats and monkeys

Feeding of raw and heated soy flour (RSF & HSF) for 4 or 8 weeks to weanling swine was compared with feeding of casein diets. There was essentially no growth in pigs fed RSF, whereas body weight increased 4.5 fold over 8 weeks with either casein or HSF. No change in pancrease weight as percent body weight was found in any group in either experiment. Both total pancreatic RNA and DNA were decreased in RSF groups, although RNA and DNA/mg pancreas were increased by the RSF diets. Total pancreatic protein, as well as protein/mg, were decreased in RSF-fed pigs compared to casein and HSF. Pancreatic trypsin, chymotrypsin, lipase, and amylase, and fecal trypsin were decreased in RSF-fed pigs to varying degrees compared to either HSF or casein. HSF and casein performance was equivalent in all parameters measured. Results of these experiments are compared with results of feeding similar diets to rats and monkeys.     

Interaction of Dietary Fat and Soybean Isolate (SBI) on Azaserine-induced Pancreatic Carcinogenesis

Raw soybean flour contains high levels of both soybean trypsin inhibitor and unsaturated fat. The chronic ingestion of this flour enhances pancreatic carcinogenesis in rats that have been treated with pancreatic carcinogens. In these studies we have examined the separate and combined effects of trypsin inhibitor in purified soybean isolate (SBI) and a high level of unsaturated fat fed during the postinitiation phase of pancreatic carcinogenesis. Male Wistar rats were injected with the known pancreatic carcinogen azaserine (30 mg/kg, ip) 14 days after birth and were weaned to the test diets. No further exposure to carcinogen occurred. Dietary composition (% by weight) of fat (corn oil) was either 5%, or for the high fat group 20%. Dietary protein (20% in all diets) was supplied as either casein, or heated or raw SBI. All diets were fedad libitum for the postinitiation phase. The number and size of transections of azaserine-induced putative, preneoplastic foci and the volume of pancreas occupied by these foci were determined by light microscopy. High fat diet increased the number and transectional size of the azaserine-induced foci as compared to the control (5% corn oil) group. The volume of pancreas occupied by foci doubled. Heated SBI had no effect upon the foci. Raw SBI, however, significantly increased the number and size of foci. The combination of raw SBI plus the high fat diet elicited an increase in focal growth that was greater than that produced by either component alone. These observations indicated that both trypsin inhibitor and the high unsaturated fat content of raw soy flour contribute to the enhancement or promotion of pancreatic carcinogenesis.     

Influence of dietary soybean trypsin inhibitors and DL-ethionine on sulfur amino acid adequacy of diets for young rats

Weanling male Wistar rats were fed 20% protein diets based on casein or either of two combinations of soy protein isolate and ground raw soy providing three levels of soybean trypsin inhibitors (SBTI; 0,448 and 808 mg of trypsin inhibited per 100g of diet respectively). DL-ethionine was included at three levels (0,0.05% and 0.10%) with each level of SBTI. After 4, 8 and 12 weeks ofad libitum feeding, diets containing SBTI without DL-ethionine were associated with decreases in weight gain, feed efficiency, serum cholesterol and serum urea nitrogen. Higher levels of triglycerides, glutamate pyruvate transaminase (SGPT) and altered serum free amino acid levels were also found. Increased dietary levels of DL-ethionine also resulted in deficits in growth and feed efficiency, decreased serum cholesterol, increased SGPT and similar alterations in serum free amino acids. Combination of dietary SBTI with DL-ethionine resulted in even greater growth deficits and serum cholesterol decreases as well as increases in SGPT and serum triglycerides and changes in serum free amino acid levels. Methionine deficiency in the young rats fed SBTI and DL-ethionine was indicated by the changes in serum amino acids and growth deficits. Moderation of some effects over the 12 week test period suggested decreased methionine requirements in the older rats.     

Long-term consumption of beef extended with soy protein by men,women and children: II. Effects on iron status

The iron status of men, women and children consuming beef extended with soy protein was evaluated by measuring serum ferritin and clinical parameters of iron status during a six-month study. Fifty-two families (245 participants) were randomly assigned to consume, for 180 days, 1 of 7 beef products: all beef, beef extended with either soy isolate, soy concentrate or soy flour (20% reconstituted soy product, 80% beef), or beef extended with each of the three soy products fortified with 60 mg Fe and 25 mg Zn/100 g protein. The beef product was consumed by the subjects as their principal source of protein for 1 meal a day (children 1–18 yr) or 1–2 meals a day (11 per week; adult men and women). A control group consumed their usual self-selected diets. No evidence was found that consumption of beef extended with soy protein deleteriously affected the iron status of men, women or children. Consumption of beef extended with soy protein, at the levels used in this study, by military men and women and by school lunch participants would not appear to impose a risk in these population groups.     

Organic and Genetically Modified Soybean Diets: Consequences in Growth and in Hematological Indicators of Aged Rats

The aim of this study was to evaluate the protein quality of organic and genetically modified soy by feeding specific diets to rats. Three groups of Wistar rats (n = 10) were used, and each group was named according to the food that they ate. There was an organic soy group (OG), a genetically modified soy group (GG), and a control group (CG). All animals received water and diet ad libitum for 455 days. At the end of this period, the weight of the GG group was the same as that of the OG, and both were higher than CG. Protein intake was similar for the OG and GG, which were significantly lower (p < 0.0005) than the CG. The growth rate (GR) of the rats, albumin levels, and total levels of serum protein were comparable for all groups. Hematocrit (p < 0.04) and hemoglobin (p < 0.03) for the OG and GG were less than the CG. Although the OG and GG demonstrated reduced hematocrit and hemoglobin, both types of soy were utilized in a way similar to casein. This result suggests that the protein quality of soy is parallel to the standard protein casein in terms of growth promotion but not hematological indicators.     

Long-term consumption of beef extended with soy protein by men,women and children: I. Study design,nutrient intakes,and serum zinc levels

The effect of long-term consumption, under practical conditions, of beef extended with soy protein on serum zinc levels of men, women and children was studied. Fifty-two families were randomly assigned to consume for 180 days 1 of 7 beef products: all beef, beef extended with either soy isolate, soy concentrate or soy flour (20% reconstituted soy product, 80% beef), or beef extended with each of the 3 soy products fortified with 60 mg Fe and 25 mg Zn/100g protein. The beef product was consumed by the subjects as their principal source of protein for 1 meal a day (children 1–8 yrs) or 1–2 meals a day (11 per week) for adult men and women. A control group consumed their usual self-selected diet. Diet records (4-day) of all foods and beverages consumed by the subjects were obtained pre-study and 63±7 and 126±7 days for calculation of nutrient intakes. No deleterious effects were found on the serum zinc levels when subjects consumed beef extended with soy protein for 180 days.Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture, and does not imply its approval to the exclusion of other products that may also be suitable.     

Comparative effects of feeding a protease inhibitor-enriched potato protein concentrate and soy flour to rats

Rats were fed casein diets to which had been added a potato protein concentrate (PPC) at a level which provided trypsin- and chymotrypsin-inhibitor activities comparable to that present at the same level (10%) of protein derived from raw soy flour. Control groups consisted of the same diets in which the unheated PPC and raw soy flour were replaced by their heated counterparts in which over 90% of the inhibitor activities had been destroyed. After 5 weeks the following measurements were made: (1) growth, (2) PER, (3) pancreas weight, (4) protein digestibility, (5) a peptide test that reflects pancreatic function, and (6) enzyme activities (trypsin, chymotrypsin, amylase, and lipase) in the pancreas and small intestine. PPC was found to produce the same general pattern of effects as raw soy flour, namely, an inhibition of growth, a decrease in PER and protein digestibility, an increase in the size of the pancreas, an increase in the recovery of p-aminobenzoic acid in the peptide test, and a depletion of enzyme in the pancreas with concomitant increases in the intestines (except for trypsin). PPC was somewhat more effective than raw soy flour in causing an increase in trypsin and chymotrypsin activities in the faces.     

Influence of sulfur-amino acid content variation in plant vs animal protein on serum and tissue lipids in rats

The objective of the study was to determine the effects of methionine and cysteine supplementation of soy protein isolate and casein on serum and tissue lipid levels in rats. Sixty male, weanling, Wistar-Kyoto rats were fed two sources of protein (casein or soy protein isolate) and three variations of sulfur-amino acid supplementation (none, methionine, or cysteine). At this level of protein intake (10% by weight), rats fed soy-based diets had similar serum lipid concentrations than rats fed casein-based diets. Choline was not added to the diet in order to be able to assess independent influences of methionine and cysteine on lipid metabolism. Overall, serum lipid values were greater in rats fed proteins supplemented with methionine while the addition of cysteine produced lower lipid levels. Liver lipid concentrations were increased tremendously upon cysteine supplementation of soy protein isolate. Protein quality, as determined by protein efficiency ratio, was improved by supplementation of either sulfur-amino acid; however, methionine had the greatest effect. Results indicate that the sulfur-amino acids influence lipid metabolism in the absence of dietary choline. The mechanism by which this occurs is not known.     

Relationship between dietary fiber levels and protein digestibility in selected foods as determined in rats

Samples of 15 food products and feces obtained by feeding them to rats were analysed for dietary fiber fractions. The food products were added as the sole source of protein in 8% protein diets, making up 8.8–51.6% of the diets. Diets were supplemented with 0.54–5.00% purified cellulose to make them more comparable in total fiber. Fiber analyses of food products revealed that the protein sources provided 0.06–7.27% total dietary fiber. The true protein digestibility in rats was negatively correlated with the total food fiber level (r=–0.69,P<0.01) or with the food cellulose level (r=–0.82,P<0.01) but it was positively correlated (r=+0.81,P<0.01) with the purified cellulose level. No relationship was found between protein digestibility and fiber fermentability. Results indicate that several food fiber fractions and possibly associated substances influenced protein digestibility. Purified cellulose did not have the same physiological behavior as food cellulose from the viewpoint of protein digestibility and fiber fermentability.     

Protein and starch digestibilities and mineral availability of products developed from potato,soy and corn flour

A technique for development of potato flour was standardized. Five products viz. cake, biscuit, weaning food, panjiri and ladoo were prepared incorporating potato flour, defatted soy flour and corn flour. Baking and roasting were the major processing techniques employed for the development of these products. Protein, ash and fat contents of potato flour were almost similar to those of raw potatoes. Significant differences in protein, ash and fat contents of all the products were observed. Protein and starch digestibility of potato flour was significantly higher than that of raw potatoes. Protein digestibility increased by 12 to 17 percent on baking or roasting of products. Processed products had significantly higher starch digestibility and mineral availability compared to raw products. Thus, it can be concluded that roasting and baking are effective means of improving starch and protein digestibility and mineral availability of products.     

Tempe Consumption Modulates Fecal Secondary Bile Acids, Mucins, Immunoglobulin A, Enzyme Activities, and Cecal Microflora and Organic Acids in Rats

The present study investigated the effect of dietary tempe, a fermented soy product, on the colonic environment of rats fed high-fat (HF, 30 % fat; experiment 1) or low-fat (LF, 6 % fat; experiment 2) diets. Growing male rats were fed the experimental diets with or without 25 % tempe for 21 days. Tempe consumption slightly but significantly increased the growth of rats fed both the HF and LF diets (P?<?0.05). With both the HF and LF diets, dietary tempe markedly reduced a harmful fecal secondary bile acid, lithocholic acid (a risk factor of colon cancer) (P?<?0.05), and markedly elevated fecal mucins (indices of intestinal barrier function) and immunoglobulin A (IgA, an index of intestinal immune function) (P?<?0.05). With the HF diet, dietary tempe increased cecal acetate, butyrate, propionate, and succinate concentrations (P?<?0.05). Analysis of the profile of cecal microflora revealed lower Bacteroides and higher Clostridium cluster XIVa levels in the tempe group of rats fed the HF diet (P?<?0.05). Compared with the control group, the fecal activity of β-glucosidase was markedly higher in the tempe group (P?<?0.05), while that of urease was lower (P?<?0.05) with both the HF and LF diets. The present results suggest that tempe consumption modulates the colonic environment in rats.     

Effects of amylose and wheat bran on the levels of blood serum urea nitrogen (BUN), other blood parameters,growth and fecal characteristics in rats

A basal diet (containing 20% soybean protein isolate) was supplemented with limiting amino acids (AA, methionine, threonine and tryptophan); wheat bran (WB, 24.3%); amylose (AM, 10%) or a combination of AA and WB or AM to investigate their effects on the levels of blood serum urea nitrogen (BUN), other blood parameters, growth and fecal characteristics in young rats. After 3 weeks feeding, supplementation of the basal diet with AA or WB resulted in improved growth (gain/food ratio) while the addition of AM had no effect on rat growth. Addition of WB to the basal diet had no effect on BUN while the addition of AA and AM caused about 20 and 12% reduction in BUN values, respectively. Rats fed the AA-AM diet had the lowest BUN values (42% lower than those fed the basal diet). Fresh volume, fresh weight, moisture and dry weight of feces in rats fed the WB diets were about four to five fold higher than in those fed the AM diets. The results suggested that BUN may not be a good predictor of growth and protein quality in diets containing AM or a source of dietary fiber (WB).     

Commercial Soy Protein Ingredients as Isoflavone Sources for Functional Foods

The knowledge of the contents and profile of isoflavones present in soy protein ingredients, as well as the effect of industrial processing, is important for the development of functional foods rich in these compounds. The results obtained here showed that the total isoflavone content varied significantly among products. For defatted and whole soy flours the total isoflavone content ranged from 120 to 340 mg/100 g, for soy protein isolates from 88 to 164 mg/100 g, and for commercial textured soy proteins, from 66 to 183 mg/100 g (wet basis, expressed as aglycones). The highest isoflavone content was found for soy hypocotyl flours, from 542 to 851 mg/100 g. Compared to hypocotyl and whole and defatted flours, soy ingredients presented a decrease of malonylglycosides and deesterified β-glycosides with a significant increase in the percentage of aglycones, mainly for soy fibers (65–76%). While defatting was shown to cause isoflavone concentration without altering conjugation, extrusion process caused destruction of isoflavones and a significant increase in the amount of acetylglycosides, but this effect was less intense for the concentrates. From the results obtained it can be concluded that differences in isoflavone concentration and profile may be related to oscillations in the isoflavone content present in the raw material and to the type of processing.     

Hypocholesterolemic effects of dietary soybean vs. casein proteins in a crossed over diets in rat

To investigate the effects of dietary proteins on the level of serum total cholesterol (TC), triglyceride (TG) and high density lipoprotein (HDL), 32 male Wistar rats were randomly divided in control and 3 experimental groups (El, E2 and E3). The feeding regimes of rats were as follow: control, standard diet; E1, a cholesterol free diet containing 20% soybean protein; E2, a cholesterol free diet containing 20% casein and E3, a cholesterol free diet containing 10% soybean protein and 10% casein. The experimental period was 11 weeks but at the end of 7th week the diets of E1 and E2 groups were crossed over for the next 4 weeks. Blood samples were collected weekly, via the ophthalmic sinus and the serum levels TC, TG and HDL were measured. In comparison with control group, the results show that at the end of 7th week TC levels in E1 and E2 groups were significantly (p<0.05) increased while HDL level unchanged and the TC value of E2 was bigger (not significant) than E1. However by crossing over the diets, the TC level was significantly (p<0.05) diminished in E2 while TG value remarkably (p<0.05) increased. These results indicate that soybean protein may insert its hypocholesterolemic effect in hypercholestrolemic condition than in normolipidemic condition.     

Nutrient composition and biological evaluation of mesta (Hibiscus sabdariffa) seeds

Two varieties of mesta (Hibiscus sabdariffa) seeds were analysed for their proximate composition. Their protein (18.8–22.3%), fat (19.1–22.8%) and dietary fiber (39.5–42.6%) contents were found to be high. The seeds were found to be a good source of minerals like phosphorus, magnesium and calcium. Their lysine and tryptophan contents were also high. Sulphur containing amino acids were limiting in this seed protein and the chemical score of mesta seed protein was 40 and 57 for AMV-2 and Bhimili-1 varieties respectively. Mesta seed oil is rich in unsaturated fatty acids (70%), of which linoleic acid constituted 44%. Weanling rats were fed with 10% mesta seed protein before and after cooking for 4 weeks. Food intake of animals receiving raw mesta seed diets was significantly lower than those receiving cooked mesta seed diets as well as the casein control diet. Protein and dry matter digestibilities of raw and cooked mesta seed diets were lower than that of casein control diet. Cooking improved the food intake, gain in body weight, dry matter and protein digestibility of mesta seed diets. PER and NPU of cooked mesta seed diets were significantly higher than the corresponding raw diets. These results indicate that cooked mesta seed protein is of relatively good quality.     

Commercial Breakfast Cereals Available in Mexican Markets and their Contribution in Dietary Fiber, β-Glucans and Protein Quality by Rat Bioassays

The beneficial effect of dietary fiber (DF) consumption has long been recognized. The global economy and open market trade policies have increased the availability of food products in Mexican markets, resulting in a wide variety of ready-to-eat commercial breakfast cereals classified as ‘high fiber’. This research was aimed to evaluate the total dietary fiber contents, its fractions (soluble and insoluble) and β-glucan in 13 commercial ‘high-fiber’ breakfast cereals, as well as to evaluate their protein quality by rat bioassays. Commercial ‘high-fiber’ breakfast cereals had 7.42–39.82 % insoluble dietary fiber, 2.53–12.85 % soluble dietary fiber, and 0.45–4.96 % β-glucan. These ready-to-eat commercial ‘high-fiber’ breakfast cereals differed significantly in their total dietary fiber, their soluble and insoluble DF fractions, and also in their β-glucan contents. When supplied as experimental diets, in 14-day rat feeding trials, the ‘high-fiber’ breakfast cereals showed an adverse effect on the % N digestibility but protein utilization, as measured as net protein ratio (NPR), was not significantly affected. The consumption of these commercial breakfast cereals, especially those made of oats as the basic ingredient, is highly recommended, since these products, being a concentrated source of dietary fiber, do not affect their protein quality.     

Effect of dietary rosemary extract on cell-mediated immunity of young rats.

The impact of rosemary extract on splenic mononuclear cell proliferation was determined. Weanling male Sprague-Dawley rats were fed diets containing 0, 100, 200 or 400 ppm rosemary extract or 400 ppm butylated hydroxytoluene (BHT) in combination with 10 or 20% casein enriched diets for 8 weeks. Splenic mononuclear cells were isolated from these animals and mitogenic response to Concanavalin A (Con A), Phytohemagglutinin (PHA) and lipopolysaccharide was determined. Con A and PHA-stimulated proliferation of spleen cells from rats fed 10% casein and 200 ppm rosemary extract was significantly higher than that of cells from the corresponding control animals. However, other levels of rosemary at 10% dietary casein or rosemary at any concentration fed along with 20% dietary casein had no impact on the mitogenic stimulation of splenic mononuclear cells. Thus, these results suggest that the use of rosemary might not have a generalized immunoenhancing effect, and will probably be effective in some stressed conditions, such as protein or antioxidant deficiency.