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 combinatiion 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.     

Tissue-specific enhancement of xenobiotic detoxification enzymes in mice by dietary rosemary extract

Plant foods contain nutritive and minor, nonnutritive components capable of inhibiting experimental carcinogenesis. Many of these cancer-protective extracts act by enhancing the activities of enzymes that can detoxify reactive substances. In the present study an extract of the spice plant rosemary was fed at concentrations of 0.3% and 0.6% (by weight) for 4 weeks to female A/J mice prior to determination of the activities of the detoxification enzymes glutathione-S-transferase (GST) and NAD(P)H: quinone reductase (QR) in lung, liver and stomach. Liver activities of GST and QR, and stomach GST activity were significantly increased in animals fed diets containing rosemary extract. However, diets supplemented with rosemary extract did not affect lung GST and QR activities. These results indicate that components of rosemary extract have the potential to protect mouse liver and stomach from carcinogenic or toxic agents.     

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.     

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.     

Partial replacement of dietary casein with soy protein isolate can reduce the severity of retinoid-induced hypertriglyceridemia

Previous research carried out in an animal model of retinoid-induced hypertriglyceridemia – rats fet a 13-cis retinoic acid (13cRA)-containing diet having casein as the protein source – has demonstrated that the complete replacement of dietary casein with soy protein isolate (SPI) can decrease the severity of this condition. In this study, the effect of partially replacing dietary casein with SPI was investigated. Five groups of male Fischer 344 rats were used in a 14-day study, with two groups being fed diets having casein as the protein source, without or with 13cRA (groups A and B, respectively), and three groups being fed 13cRA-containing diets in which SPI was used to bring about the isonitrogenous replacement of 25, 50, or 100% of the casein in the formula for the diet used for group B (groups C-E, respectively). Serum triglyceride concentration for group B was significantly different ( p < 0.05) from that of groups A, D, and E (5.41 vs 2.62, 4.04, and 2.66 mmol/l, respectively). Serum cholesterol concentrations for groups D and E were significantly lower ( p < 0.05) than for groups A and B (1.63 and 1.60 vs 2.00 and 2.14 mmol/l, respectively). Thus, the isonitrogenous replacement of 50% of dietary casein with SPI can reduce the severity of retinoid-induced hypertriglyceridemia while decreasing the serum concentration of cholesterol.     

Iodine metabolism and tissue mineral levels in rats fed rapeseed protein concentrates supplemented with zinc

A 16-week feeding test of Tower (Brassica napus) and Span (Brassica campestris) rapeseed protein concentrates (RPC) was carried out on male and female Charles River CD (Sprague-Dawley) weanling rats. The diets contained 10% or 20% protein (N × 6.25) from each RPC and were supplemented with 150 mg zinc/kg. The control groups received a purified diet with casein or a laboratory chow diet. The results of the test diet groups did not reveal any adverse effect due to RPC on thyroid metabolism or the metabolism of zinc iron, copper, manganese, calcium, phosphorus, and magnesium. Nephrocalcinosis was observed in all female rats that were fed the test diets and the casein control diet, and therefore it could not be attributed to dietary RPC.Bureau of Nutritional Sciences, no. 120. Food Research Institute, no. 448.     

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.     

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.     

The cholesterol-lowering property of soybeans fed to rats is related to the fasting duration

Numerous factors affect plasma cholesterol and, therefore, representpotential interferences with the specific effects of diet on lipid metabolism.After casein intake, serum cholesterol levels depend on many experimentalconditions in rats, such as the type of casein, the presence or absence ofcholesterol in the diet, the age and strain of rat, the diet composition, thenutritional status. The effect of fasting duration on selected parameters wasstudied in adult male Wistar rats fed a 20% casein and 1% cholesterol-based diet. No changes were observed in total serum, lipoproteins-B andHDL cholesterol and in triglyceride values after an 8 h-fast compared tonon-fasted animals. A significant decrease was induced in total andlipoproteins-B cholesterol with a 12 h-fast. A prolonged fast resulted ina significant decrease in these parameters and in triglycerides and inHDL-cholesterol levels.Fasted animals fed heated soybean meal instead of casein did not display amarked decrease in serum lipids. It appeared that the fasting effect wasmore marked in animals fed casein than in animals fed soybeans. Theseresults showed the importance of experimental conditions such as nutritional status ofanimals when blood samples are taken in studies of the hypocholesterolemiceffect of soy protein compared to casein.     

Growth,blood chemistry and histology of rats fed zincsupplemented rapeseed protein concentrates

Two rapeseed protein concentrates (RPC) supplemented with zinc oxide were fed to male and female rats at two levels providing approximately half or all of the protein in a purified basal diet containing 20% protein (N×6.25). Control groups were given either a purified basal diet with casein or a laboratory chow diet. After 16 weeks, there were no significant (P<0.05) differences in body weight except the male rats fed the casein control diet had higher body weights than did other male rats. No significant differences were observed in organ weights including thyroids, hematology, bone marrow differential counts, serum glucose, urea nitrogen, total protein, albumin/globulin ratio, cholesterol, transaminases or alkaline phosphatase. The levels of serum vitamins A and E of females fed some of the RPC diets were less than in those fed the caseincontrol diet, but equal to those in females fed laboratory chow diet. Females fed either RPC or casein in the semi-purified diets, which contained zinc oxide, exhibited kidney calcification.Bureau of Nutritional Sciences, No. 112; Food Research Institute, No. 420.     

Bioavailability of tryptophan in selected foods by rat growth assay

Tryptophan bioavailabilities were estimated in 16 protein sources using 10 day rat growth assays with casein as the reference protein. Growth responses of rats fed test food diets were compared to growth responses of rats fed basal diets with graded levels of tryptophan ranging from 50 to 100 mg of tryptophan/100 g diet. Estimates of tryptophan availabilities were 85–100% for all products except whole wheat cereal (73%) and pinto beans (59%). Results of a previous study on lysine availability indicated that poor response to pinto beans was due either to poor digestibility or to the presence of some unidentified growth inhibitor.     

Effect of supplementation of blue green alga (Spirulina) on outcome of pregnancy in rats

To study the supplementary effect of Spirulina, pregnant rats were fed 5 different kinds of diets (casein, Spirulina, wheat gluten, Spirulina + wheat gluten, Spirulina-without additional vitamins and minerals), each providing 22% protein during the period of pregnancy. The outcome of pregnancy was assessed from litter and dams' weight and litter size. Maternal weight gain was found to be maximum with Spirulina + wheat gluten and least with the wheat gluten diet. Rats receiving Spirulina containing diets produced significantly (p<0.05) higher litter size than those receiving casein and wheat gluten. In spite of having higher litter size, Spirulina containing diet groups produced pups with birth weights comparable to those of casein. Spirulina appears to be a good dietary supplement during pregnancy.     

Effects of Blackcurrant and Dietary Fibers on Large Intestinal Health Biomarkers in Rats

This study examined the effects of anthocyanin-rich blackcurrant extract and dietary fibers individually and their combinations on biomarkers of large intestinal health in rats. After six weeks of feeding, rats fed diets with blackcurrant gained significantly less body weight and reduced their food intake resulting in a lower food efficiency compared with those rats fed control diets. Combining dietary fiber (apple or broccoli) with blackcurrant in the diet was more effective in reducing the body weight gain and food intake. Cecal bacterial populations and short-chain fatty acids differed between the experimental diets. Blackcurrants significantly altered the bacterial populations by increasing the abundance of Bacteroides-Prevotella-Porphyromonas group and Lactobacillus spp., while decreasing the abundance of Bifidobacterium spp. and Clostridium perfringens. Propionic acid concentrations were increased by the diets with blackcurrant. Butyric acid concentrations were increased by dietary fiber supplementation. Dietary fiber increased the number of goblet cells in the colon. Diets with blackcurrant were more effective in altering the biomarkers of large intestinal health than those without blackcurrant.     

Influence of Dietary Avocado on Gut Health in Rats

This study investigated the impact of diets containing various levels of avocado (5, 10 and 15%) on gut health in rats fed for six weeks. Avocado-fed rats had significantly higher food intakes while their body weights remained similar to the control diet-fed rats. No significant changes in intestinal bacterial populations (ileum, cecum and colon) were found in rats fed avocado diets compared to the control diet. Ileum and colon tissues of rats fed avocado diets showed significantly higher expression of genes (β-defensin 1, mucin 3 or mucin 4) and a greater number of mucin-producing goblet cells in the colon. The percentage of avocado in the diet had varying effects in altering the biomarkers, whereby diet containing 15% avocado was the more effective diet. This study delivers new knowledge on the role of avocado on gut health in rats.     

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.     

Effect of turmeric on xenobiotic metabolising enzymes

Diet contains several substances capable of inhibiting chemical carcinogenesis. It is known that such inhibitors may either act directly by scavenging the reactive substances or indirectly by promoting mechanisms which enhance detoxification. Turmeric which contains curcumin both in vitro and in vivo is an active antimutagen. Studies were therefore conducted to evaluate the effects of turmeric on xenobiotic metabolising enzymes in hepatic tissue of rats fed turmeric ranging from 0.5–10% in the diet. Enzymes such as aryl hydrocarbon hydroxylase, UDP glucuronyl transferase and glutathione-S-transferase were assayed after four weeks of turmeric fed diets. No significant differences were seen in the activating enzyme AHH. However, UDPGT was significantly elevated in rats fed 10% turmeric while GSHT registered a significant increase in 5 and 10% turmeric fed diet as compared to controls and 0.5–1.0% turmeric fed animals. The results suggest that turmeric may increase detoxification systems in addition to its anti-oxidant properties. Curcumin perhaps is the active principle in turmeric. Turmeric used widely as a spice would probably mitigate the effects of several dietary carcinogens.     

Determination of iron absorption by rat bioassay. Evaluation of methods of dosing59Fe on radioiron absorption from plant diets

We studied the potential of an animal model to predict nonheme iron absorption in humans and tested a feasible and easy technique of dosing⁵⁹Fe to evaluate bioavailability of dietary nonheme iron. Plant diets containing about 20 ppm or 75 ppm iron were the nonheme iron sources with ferrous sulfate (75 ppm) as a reference. Radioiron was administered by (a) gavaging in water, 1 h after a meal; (b) mixing with the meal; and (c) making a slurry with the meal and gavaging. No significant differences were found (P0.05) in⁵⁹Fe absorption among the three methods of administering radioiron. Absorption of⁵⁹Fe was similar to apparent iron absorption for all diets tested, whether the animals consumed the same diet as that of the test meal or a different diet. A high correlation (r=0.88) obtained between the apparent iron absorption and⁵⁹Fe iron absorption for different dosing techniques, indicates that extrinsic iron tag administered by any of these methods is valid to measure nonheme iron absorption. Apparent iron absorption values determined by rats fed 20 ppm or 75 ppm dietary iron from plant sources were similar to reported values for humans. It is concluded that the normal adult rat is an appropriate model to study iron bioavailability of human foods when iron status, maturity, iron intake relative to requirement, and method of measurement are similar to the human situation.     

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

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.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

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).     

The bioavailability of magnesium from Wakame ( Undaria pinnatifida) and Hijiki (Hijikia fusiforme) and the effect of alginic acid on magnesium utilization of rats

The bioavailability of magnesium from Wakame and Hijiki, and the effects of alginic acid on absorption of dietary magnesium were examined in five groups of rats fed either control, Wakame, Hijiki, AW (containing the same amount of alginate as in the Wakame) and AH (containing the same amount of alginate as in the Hijiki) diets, and animals fed a low magnesium diet (LMg) (twentieth amount of magnesium in the original mineral mixtures as the control). Food intake and body weight gain were decreased by adding sodium alginate to the diets. A large amount of calcium accumulated only in the kidneys of the rats fed the LMg diet. Serum magnesium concentration decreased only in the LMg group. The magnesium content in the defatted left femurs did not differ between the control and Wakame fed animals and also among the animals eating Wakame, Hijiki and AW diets. The breaking force of the right femurs did not differ among all the groups except the LMg group. The ratio of apparent magnesium absorption (%) of the control, LMg, Wakame, Hijiki, AW and AH groups was 82.2, 72.7, 66.9, 50.8, 69.3 and 54.2 in the first experimental period, and was 75.3, 52.1, 57.7, 46.9, 62.6 and 60.5 in the second experimental period, respectively. It was clear that the bioavailability of magnesium in the Wakame fed rats was higher than in those eating the Hijiki. Large amounts of sodium alginate lowered magnesium absorption from the diet.