Ingestion of insoluble dietary fibre increased zinc and iron absorption and restored growth rate and zinc absorption suppressed by dietary phytate in rats

We examined the effects of ingestion of five types of insoluble fibre on growth and Zn absorption in rats fed a marginally Zn-deficient diet (6.75 mg (0.103 mmol) Zn/kg diet) with or without added sodium phytate (12.6 mmol/kg diet). The types of insoluble fibre tested were corn husks, watermelon skin, yam-bean root (Pachyrhizus erosus) and pineapple core, and cellulose was used as a control (100 g/kg diet). Body-weight gain in the cellulose groups was suppressed by 57 % by feeding phytate. Body-weight gain in phytate-fed rats was 80 % greater in the watermelon skin fibre and yam-bean root fibre group than that in the cellulose group. Zn absorption ratio in the cellulose groups was lowered by 46 and 70 % in the first (days 7-10) and second (days 16-19) measurement periods with feeding phytate. In the rats fed the phytate-containing diets, Zn absorption ratio in the watermelon skin, yam-bean root and pineapple core fibre groups was 140, 80 and 54 % higher respectively than that in the cellulose group, in the second period. Fe absorption was not suppressed by phytate, however, feeding of these three types of fibre promoted Fe absorption in rats fed phytate-free diets. The concentration of soluble Zn in the caecal contents in the watermelon skin fibre or yam-bean root fibre groups was identical to that in the control group in spite of a higher short-chain fatty acid concentration and lower pH in the caecum. These findings indicate that ingestion of these types of insoluble fibre recovered the growth and Zn absorption suppressed by feeding a high level of phytate, and factors other than caecal fermentation may also be involved in this effect of insoluble fibre.     

Role of copper in the regulation and accumulation of superoxide dismutase and metallothionein in rat liver

Weanling male Sprague-Dawley rats were fed one of four diets that varied in Cu, Zn or Cd content. To the control diet (I) Cu, Zn and Cd were added at 10, 100 and 0 mg/kg diet, respectively. Diets II and III also contained 10 mg/kg of dietary Cu, except that Zn was elevated to 1000 mg/kg for diet II, or Cd was added at 10 mg/kg for diet III. Diet IV was deficient in Cu (less than 1 mg/kg) with Zn at 100 mg/kg and no added Cd. At wk 6 postweaning, half of the rats fed diets I and IV were injected once with Cd acetate (5 mg Cd/kg body weight). The immediate response to Cd injection was an increase in metallothionein accumulation (three- to fourfold) and in Cu,Zn superoxide dismutase (SOD) accumulation (1.2- to 1.5-fold) in liver. SOD was estimated in an ELISA. These responses were not influenced by a change in Cu status (I vs. IV). However, in functional assays, SOD enzymatic activity was about half that of the control values. In this regard, SOD appears to be given high priority with respect to the utilization of cellular Cu, i.e., a 10-fold reduction in hepatic Cu only resulted in a twofold reduction in SOD activity and the amount of apoenzyme remained at normal levels.     

The effects of phytate on intestinal absorption and secretion of zinc, and whole-body retention of Zn, copper, iron and manganese in rats.

1. The inclusion of phytate (10 g/kg) in a purified diet containing zinc (15 mg/kg) fed to young male rats significantly reduced growth rate and food intake, and promoted a cyclic pattern of food intake characteristic of an uncomplicated Zn deficiency. The decreased growth rate could be accounted for by the reduced food consumption. 2. Rats maintained on a Zn-deficient diet (0.5 mg Zn/kg) were found to have a cyclic pattern of food intake and a very slight weight gain. The addition of phytate (10 g/kg) to the Zn-deficient diet promoted a net loss of mean body-weight. 3. Rats maintained on the Zn-supplemented diet containing phytate excreted significantly more Zn in their faeces than either pair-fed or ad lib.-fed control rats. Rats given the Zn-deficient diet supplemented with phytate excreted more Zn in their faeces than Zn-deficient control rats. 4. Dietary phytate significantly reduced the average daily accumulation (mug/d) and whole-body retention (relative to dietary intake) of iron, copper, manganese and Zn, whether or not the diet was supplemented with Zn. 5. The addition of phytate to the lumen fluid of ligated loops of rat duodenum maintained in situ significantly inhibited 65Zn absorption, compared with the control systems without added phytate. 6. Other studies using ligated duodenal and ileal loops indicated that Zn is secreted into the gut lumen and approximately one-third of this is normally reabsorbed. Recycling of endogenous Zn may be a significant process in the over-all body economy of this trace element. 7. The absorption of 65Zn added to the diet was significantly reduced by dietary phytate. Dietary phytate also reduced the biological half-life of body 65Zn from 61 to 211 h post-administration, possibly by inhibiting reabsorption of endogenous 65Zn and thus promoting a more rapid loss from the body.     

Comparison of chemical, histomorphometric, and absorptiometric analyses of bones of growing rats subjected to dietary calcium stress

Absorptiometric, histomorphometric, and chemical analyses of bones from growing rats fed diets with low (0.2%, w/w), marginal (0.4%, w/w), or adequate (0.8%, w/w) calcium (Ca) content with or without phytate were compared. Phytate was added to each diet in a molar ratio of 19:1 to calcium. Male weanling Sprague-Dawley rats were fed one of the six diets for 8 weeks. At the end of 8 weeks, rats were killed, and mandibles, femurs, and tibias were removed. Bone density profiles were determined on the mandibles and femurs using single photon absorptiometry. Femurs were also used for calcium and phosphorus analyses. Tibias were used for histomorphometric analyses. Bone density of the femurs and mandibles increased as dietary Ca increased. The only effect of phytate addition measured was in the 0.8% calcium diet, where density was lower in rats fed the phytate-containing 0.8% calcium diet. Femur calcium concentration also increased as dietary Ca increased and was unaffected by addition of phytate. Femur phosphorus concentration was unaffected by dietary Ca levels but was increased by 10% when phytate was added to the diet. Bone density values were highly correlated with bone calcium and phosphorus levels (r = 0.94). Rats fed the 0.2% calcium diets had 20% lower mineralized bone area and 20% larger medullary cavity area than rats fed the other diets. Bone densitometry appears to be useful for determining changes in bone occurring in growing rats fed low, marginal, and adequate levels of dietary Ca. Bone density values also correlated well with chemically determined calcium and phosphorus concentrations and with histomorphometric data.     

Cadmium absorption and retention by rats fed durum wheat (Triticum turgidum L. var. durum) grain

A whole-body radioassay procedure was used to assess the retention and apparent absorption by rats of Cd in kernels of durum wheat (Triticum turgidum L. var. durum) harvested from plants grown hydroponically in 109Cd-labelled nutrient solution. Wholegrain wheat, containing 5 micromol Cd (570 microg)/kg dry weight labelled intrinsically with 109Cd, was incorporated into test meals fed to rats that had been maintained on diets containing marginally adequate, adequate or surplus levels of Zn (0.12 mmol (8 mg), 0.43 mmol (28 mg) or 1.55 mmol (101 mg) Zn/kg respectively), and either 0 or 50 g durum wheat/kg. Regardless of diet, all rats consumed about 99 % of the test meal offered. In rats fed diets without wheat, initial Cd absorption averaged 7.7, 4.6 and 2.4 % of the dose when the diet contained 0.12 mmol (8 mg), 0.43 mmol (28 mg) or 1.55 mmol (101 mg) Zn/kg diet respectively. In rats fed wheat-containing diets, initial Cd absorption averaged 3.8 and 2.6 % of the dose when dietary Zn concentration was 0.12 mmol (8 mg) and 0.43 mmol (28 mg)/kg diet respectively. The amount of Cd retained in the body at 15 d postprandial was <2 % of the dose in all rats, and decreased as Zn in the diet increased. Even at 15 d postprandial, 32 to 44 % of the Cd retained in the body was still in the gastrointestinal tract. The results show that: (1) the bioavailability to rats of Cd in wholegrain durum wheat was depressed when wholegrain wheat was part of the regular diet; (2) increased intake of dietary Zn lowered Cd absorption and retention; (3) retention of Cd in the body at 15 d postprandial from diets containing adequate Zn was <1.3 %.     

Comparison of chemical,histomorphometric, and absorptiometric analyses of bones of growing rats subjected to dietary calcium stress.

Absorptiometric, histomorphometric, and chemical analyses of bones from growing rats fed diets with low (0.2%, w/w), marginal (0.4%, w/w), or adequate (0.8%, w/w) calcium (Ca) content with or without phytate were compared. Phytate was added to each diet in a molar ratio of 19:1 to calcium. Male weanling Sprague-Dawley rats were fed one of the six diets for 8 weeks. At the end of 8 weeks, rats were killed, and mandibles, femurs, and tibias were removed. Bone density profiles were determined on the mandibles and femurs using single photon absorptiometry. Femurs were also used for calcium and phosphorus analyses. Tibias were used for histomorphometric analyses. Bone density of the femurs and mandibles increased as dietary Ca increased. The only effect of phytate addition measured was in the 0.8% calcium diet, where density was lower in rats fed the phytate-containing 0.8% calcium diet. Femur calcium concentration also increased as dietary Ca increased and was unaffected by addition of phytate. Femur phosphorus concentration was unaffected by dietary Ca levels but was increased by 10% when phytate was added to the diet. Bone density values were highly correlated with bone calcium and phosphorus levels (r = 0.94). Rats fed the 0.2% calcium diets had 20% lower mineralized bone area and 20% larger medullary cavity area than rats fed the other diets. Bone densitometry appears to be useful for determining changes in bone occurring in growing rats fed low, marginal, and adequate levels of dietary Ca. Bone density values also correlated well with chemically determined calcium and phosphorus concentrations and with histomorphometric data.     

Effect of dietary phytate on bioavailability of iron

Experiments were carried out on iron-chelating capacity of phytic acid in fortified and unfortified diets of whole wheat, white flour and wheat bran using albino rats in a 10-day balance study. The results revealed that gross digestibility was highest in control diet (93.25%), least in the wheat brain diet with added ferrous sulfate (86.70%) and intermediate in other diets. Among the unfortified and fortified diets, the highest amount of phytate was present in the diet having wheat bran (2.69–4.99 mg/g). Phytate concentration of the rat faeces followed the pattern of dietary phytate content of test diets. The data indicated high values for iron (200.00–236.64 mg/g) in fortified diets while in the unfortified diets, the value was the same (16.67 mg/g) except the casein diet, which contained 69.99 mg/g of iron. Iron concentration of the fat faeces ranged 16.25–23.50 mg/g; the higher value being in the excreta of animals given fortified diets. The casein diet had iron 46.75 mg/g. The overall results showed that the phytate absorption was relatively higher in groups fed fortified diets. Iron absorption almost followed the pattern of phytate absorption. Correlation co-efficient analysis showed insignificant relationship between iron and phytate absorption indicating little effect of dietary phytate on the biovailability of iron.     

The effect of heat treatment and particle size of bran on mineral absorption in rats

The effect of heat treatment of bran on the true zinc absorption was measured using an isotope-dilution technique. A bran-based breakfast cereal (heated to 204 degrees for 40 min during manufacture) was incorporated into a semi-synthetic diet at a level of 180 g/kg. A parallel diet was formulated containing an identical weight of untreated bran from the same source plus other ingredients used to make the cereal. 2. Young male Wistar rats (mean weight 80 g) were injected intramuscularly with 65Zn to label body Zn. They were given the heat-treated- and untreated-bran diets for 9 d. During the last 6 d of this period Zn intakes and faecal and urinary Zn were measured in order to calculate apparent Zn retention. True Zn retention was measured by taking into account losses of Zn of endogenous origin (labelled with 65Zn), by measuring faecal and urinary radioactivity of endogenous origin. 3. Heat treatment of bran removed approximately one-third of the phytate, but this was not enough to improve Zn absorption from the diet. True Zn retention measured by isotope dilution was significantly higher (P less than 0.02) than apparent Zn retention measured by the conventional balance technique. 4. The hypothesis that a reduction in particle size of bran would improve mineral availability was tested by feeding coarse and milled bran (100 mg/kg diet) in a semi-synthetic diet to rats and measuring true Fe and apparent Zn absorptions. The importance of phytate was also investigated by feeding a diet containing dephytinized bran. 5. Male Wistar rats (mean weight 172 g) were given diets containing coarse, milled or dephytinized bran for 9 d. Fe and Zn intakes were measured and faeces and urine collected for Fe and Zn analysis. 6. Rhe mean (+/- SE) particle size of the bran was reduced on milling from 3.5 (+/- 1.8) to 0.2-0.5 mm. There were no differences in the fraction of Fe retained between the three groups. Particle size had a small effect on Zn retention which was marginally higher in rats on the milled-bran diet (0.126 (+/- 0.023)) than in those on the coarse-bran diet (0.087 (+/- 0.012)). Total removal of phytate had a greater effect and apparent Zn retention from the dephytinized-bran diet was significantly higher (0.182 (+/- 0.027), P less than 0.001).     

Comparative effects of wheat bran and barley husk on nutrient utilization in rats. 2. Zinc, calcium and phosphorus

1. The present work was undertaken to study comparatively the effect on mineral availability in rats of wheat bran and barley husk when supplying the same amount of dietary fibre (DF). The experiment involved a total of nine dietary treatments including a control group and two series of four groups with increasing amounts of fibre from the two sources (total DF ranging from 42 to 117 g/kg dry matter (DM]. Dietary nitrogen concentration was kept constant at 15 g N/kg DM. Zinc concentration of the diets was adjusted to the level provided by the diet with the highest wheat-bran content (21 mg/kg DM) using zinc sulphate. Other minerals were not adjusted. 2. Two experiments were performed. In Expt 1 the diets were given to 5-week-old rats during 9 d and apparent absorptions of Zn, calcium and phosphorus and the femur concentrations of Zn, Ca and P were measured. In Expt 2 the diets were given to 9-week-old rats during 12 d. Mineral concentration in femur and total and albumin-bound plasma Zn and availability of plasma Zn for enzyme reactivation were measured. 3. In the younger animals, wheat bran depressed significantly the absorption of Zn when providing 40 g DF/kg DM and absorbtion of Ca when providing 80 g DF/kg DM. Barley husk depressed significantly both the absorption of Zn and Ca already at 20 g DF/kg DM. Both fibre sources had a more negative effect on Zn than on Ca absorption. Only barley husk had a small negative effect on absorption of P. Phytate did not appear as a major factor affecting mineral absorption in barley husk. All diets containing barley husk had a very low molar ratio, phytate:Zn. 4. The age of the animals influenced the utilization of dietary minerals using femur concentration as a criterion, particularly in the case of Zn. In the younger animals the decrease in femur Zn with fibre correlated with apparent Zn absorption both with wheat bran (R2 0.986, P less than 0.01) and with barley husk (R2 0.996, P less than 0.01). In the older animals femur Zn did not change significantly with fibre. 5. In the older animals, plasma Zn, albumin-bound plasma Zn and availability of plasma Zn for enzyme reactivation were lowest with the highest addition of wheat bran.     

Effects of dietary phytate, calcium and magnesium levels on zinc bioavailability to rats

Young rats were fed diets containing 12 mg Zn/kg and varied levels of sodium phytate for 21-day ad libitum feeding periods. In experiment 1, Ca levels were 0.3, 0.5, 0.8 and 1.0%, and phytate:Zn molar ratio varied between 0 and 50. In experiment 2, Ca was maintained at 0.3%, Mg levels were 0.07, 0.22 and 0.37%, and phytate:Zn molar ratios were 0, 10, 20 and 30 at each Mg level. Major response criteria were body weight gain and tibia Zn accumulation. Weight gain was not influenced by Ca level in the absence of phytate or by phytate at 0.3% Ca; it was increasingly depressed as phytate was increased and by each increase in Ca in the presence of phytate. Total tibia Zn content was decreased at the highest Ca level in the absence of phytate; increasing the phytate progressively depressed tibia Zn at all Ca levels. Mg and phytate additions did not affect weight gain. Tibia Zn tended to be depressed by Mg and by phytate but these effects were significant only at the highest levels of the combined additions. These data corroborate and extend previously published findings on Ca and phytate effects on Zn utilization and show bone Zn accumulation to be a more sensitive criterion than weight gain in this connection. They also indicate that Mg exerts a less pronounced effect on Zn utilization in phytate-containing diets than does Ca.     

The addition of milk or yogurt to a plant-based diet increases zinc bioavailability but does not affect iron bioavailability in women

The addition of milk and milk-based products to the diets of individuals subsisting on plant-based diets was reported to have positive effects on nutritional status and functional outcomes such as growth, morbidity, and cognition. We examined the effect of the addition of milk or yogurt on the bioavailability of zinc and iron from a plant-based rural diet. The subjects were 48 Mexican women (30.9 +/- 5.7 y) who habitually consumed a plant-based diet. The women were assigned to 1 of 3 groups: 1) the typical rural Mexican diet, 2) that diet with milk added, or 3) that diet with yogurt for 13 d. Zinc absorption was measured after extrinsically labeling meals with (67)Zn and an i.v. dose of (70)Zn; iron absorption was measured by extrinsically labeling meals with (58)Fe and a reference oral dose of (57)Fe. Including milk and yogurt in the diet increased zinc absorption by 50 and 68%, respectively (P < 0.05). The 3 groups did not differ in the percentage iron absorption. The total amount of zinc absorbed was increased (P < 0.05) by 70% when milk was added to the meal and 78% when yogurt was added. The total amount of iron absorbed did not differ among the groups. The addition of milk and yogurt to a plant-based diet high in phytate increases zinc bioavailability without affecting iron bioavailability.     

The similarity between alkaline phosphatase (EC 3.1.3.1) and phytase (EC 3.1.3.8) activities in rat intestine and their importance in phytate-induced zinc deficiency.

1. The activities of alkaline phosphatase (EC 3.1.3.1) and phytase (EC 3.1.3.8) were similarly distributed in the small intestine of rats. Regional differences in activity were reflected by similar differences in the capacity of ligated intestinal segments to hydrolyse phytate in vivo. Activities were greatest in the duodenum and lowest in the terminal ileum. 2. Specific activities of both enzymes were tenfold greater in the brush border fraction of duodenal mucosa compared with entire mucosal homogenates. 3. Brush-border alkaline phosphatase and phytase activities required both magnesium and zinc ions for maximal activity. 4. Zn deficiency induced by feeding a diet low in Zn (0.5 mg Zn/kg) caused similar reductions in activity of both enzymes. 5. Zn deficiency induced by feeding diets marginally adequate in Zn (12 mg/kg) and phytate (10 g/kg) caused reductions in alkaline phosphatase, phytase activities and phytate hydrolysis in vivo. 6. It is suggested that phytase activity is a manifestation of alkaline phosphatase and the significance of this in relation to phytate-induced in Zn deficiency is discussed.     

Effect of dietary phytate on zinc homeostasis in young and elderly Korean women

BACKGROUND: Previous studies suggest that consumption of predominantly plant-based diets with high phytate content contribute to zinc deficiency by inhibiting zinc absorption. Age of the individual may also affect the ability to maintain zinc homeostasis. OBJECTIVE: This study was designed to determine the effect of dietary phytate on zinc homeostasis and to evaluate the effect of age on the capacity to maintain the zinc homeostasis with changes in dietary phytate in young and elderly Korean women. DESIGN AND METHODS: Seven healthy young women (22-24 yr) and 10 healthy elderly women (66-75 yr) were studied consecutively for 3 months in 2 metabolic periods (MP) in two different metabolic units. During MP1 the women consumed a high phytate (HP) diet (P:Zn molar ratio = 23) for 9 days. After a 10 d wash-out period at home eating their usual diets, a lower phytate diet (LP) (P:Zn molar ratio = 10) was fed in MP2 for 9 d. Phytase was added to selected foods in the high phytate diet to reduce the phytate content of the meals in the LP period. The zinc content of both diets was about 6.5 mg/d. Stable isotopes of Zn ((70)Zn) were administered intravenously on d 5 of MP 1 and 2 for measuring endogenous fecal zinc excretion. Plasma samples were also collected on d 5 for measuring plasma zinc concentrations by Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES). 24 hr urine samples were collected for 5 d and complete fecal samples were collected for 9 d after isotope administration. Fractional zinc absorption (FZA) was calculated from mass balance corrected for endogenous fecal zinc (EFZ) excretion and EFZ was determined by using an isotopic dilution technique. Isotopic ratios for FZA and EFZ were measured by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Statistical analyses were done using ANOVA. RESULTS: Both the young and elderly women were in negative zinc balance during the HP period. This was due to a significant decrease in FZA and total absorbed zinc (TAZ) with a HP diet (43 vs 22% in young women, 34 vs 20% in elderly women, p < 0.001). EFZ excretion did not differ in the young and elderly women during the LP and HP periods. Dietary phytate did not alter plasma zinc concentrations or and urinary zinc excretion in either group. CONCLUSIONS: Adjustments in zinc homeostasis with an increase in dietary phytate did not differ between young and elderly women in this study.     

Effects of dietary zinc levels, phytic acid and resistant starch on zinc bioavailability in rats

Summary Background Owing to its fermentability, it has been advocated that resistant starch (RS) has a positive effect on the absorption of minerals by increasing their solubility in the hindgut. In marginally zinc–deficient rats, the enhancement of zinc bioavailability by RS occurs mostly when the diet contains phytic acid. Aim of the study This study aims to investigate the effect of dietary zinc level and phytic acid on the cecal zinc pools and zinc bioavailability of rats fed RS. Methods Wistar rats (male, 3wk old) were divided into eight groups (n = 6), and fed diets containing either 5% cellulose (control fiber: insoluble and low fermentable) or 20 % RS (test fiber: soluble and fermentable), with or without the addition of 1% sodium phytate, at the 10 and 30 mg/kg dietary zinc levels, for 21 days. Results At 10 mg Zn/kg, RS increased femur zinc concentration only in the group receiving the phytate–containing diet, while at 30 mg Zn/kg it increased femur zinc concentration in rats fed both phytate–free and phytate–containing diets. The total content of zinc in the cecum was increased by the higher dietary zinc level and tended to be increased by the addition of phytate, which is assumed to impair zinc absorption in the small intestine. Feeding RS lowered cecal pH values, which correlated with increasing values of zinc solubility (r = –0.3471; P < 0.05). The later was, in turn, directly associated with zinc apparent absorption (r = 0.3739; P < 0.05). Conclusions The increase in zinc bioavailability by RS occurs when dietary zinc levels are adequate and/or zinc absorption is impaired in the small intestine, increasing the influx of unabsorbed zinc into the cecum and favoring the increase of zinc bioavailability when RS fermentation lowers the cecal pH.     

A model to produce pure zinc deficiency in rats and its use to demonstrate that dietary phytate increases the excretion of endogenous zinc

An early effect of zinc deficiency in rats is loss of appetite with consequent malnourishment. To obviate the need for pair-feeding, rats were fed liquid semipurified diets by gastric tube feeding. Rats tube fed a zinc-deficient diet at a daily rate of 90-110 g/kg thrived for 6-7 days and then rapidly became seriously ill. In contrast rats fed the deficient diet ad libitum stopped growing after 3 days but remained relatively healthy. Animals tube fed a zinc-replete diet or the deficient diet with subcutaneous injections of zinc grew normally and suffered no ill effects. Rats tube fed the deficient diet and supplemented parenterally with zinc excreted significantly more endogenous zinc into small intestinal luminal washings and into feces than unsupplemented rats. Sodium phytate, added to the tube feed in three groups of rats fed the deficient diet and supplemented daily with 0, 0.33 and 3.3 mg Zn/kg, significantly increased the zinc content of luminal washings in all three groups, increased fecal zinc excretion in two groups and lowered body zinc levels, estimated by femur zinc, in two groups. Tube feeding provides a means to study: 1) pure zinc deficiency without malnourishment of other nutrients; and 2) the excretion of endogenous zinc into the gastrointestinal tract.     

Influences of dietary protein levels and phytate contents on zinc requirement in rats

This study investigates whether the effects of increased dietary levels of soy protein on zinc-deficient rats are due to the dietary protein content. Rats were fed two levels of demineralized soy protein (DP) diets and two levels of egg albumin (EA) diets including 7 ppm zinc (Experiment 1). Growth was depressed in rats fed a 20% DP diet (0.43% phytate) but not in those fed a 10% EA diet containing 7 ppm zinc. Zinc concentrations in the serum, femur and kidney were lower in rats fed the 20% DP diet than those fed a 10% DP (0.20% phytate) diet, and they were also lower in rats fed the 10% EA diet than those fed a 5% EA diet. Zinc concentrations in the serum and femur of rats fed the 10% DP diet or the 20% DP diet were decreased compared with those fed the 5% EA diet or the 10% EA diet, respectively. Under zinc-deficient conditions (< 0.4 ppm Zn), the survival time shortened (Experiment 2) and the zinc concentration in the serum and femur decreased faster (Experiment 3) in rats fed the 10% EA diet compared with those fed the 5% EA diet. The survival times and time courses of these parameters show that the zinc requirements of rats increased with the dietary protein level. The increased zinc requirement of rats that accompanied increasing dietary soy protein was due to the dietary protein content as well as the dietary phytate content.     

Influences of dietary protein levels and phytate contents on zinc requirement in rats

This study investigates whether the effects of increased dietary levels of soy protein on zinc-deficient rats are due to the dietary protein content. Rats were fed two levels of demineralized soy protein (DP) diets and two levels of egg albumin (EA) diets including 7 ppm zinc (Experiment 1). Growth was depressed in rats fed a 20% DP diet (0.43% phytate) but not in those fed a 10% EA diet containing 7 ppm zinc. Zinc concentrations in the serum, femur and kidney were lower in rats fed the 20% DP diet than those fed a 10% DP (0.20% phytate) diet, and they were also lower in rats fed the 10% EA diet than those fed a 5% EA diet. Zinc concentrations in the serum and femur of rats fed the 10% DP diet or the 20% DP diet were decreased compared with those fed the 5% EA diet or the 10% EA diet, respectively. Under zinc-deficient conditions (<0.4 ppm Zn), the survival time shortened (Experiment 2) and the zinc concentration in the serum and femur decreased faster (Experiment 3) in rats fed the 10% EA diet compared with those fed the 5% EA diet. The survival times and time courses of these parameters show that the zinc requirements of rats increased with the dietary protein level. The increased zinc requirement of rats that accompanied increasing dietary soy protein was due to the dietary protein content as well as the dietary phytate content.     

Inhibitory effect of dietary soybean protein vs. casein on magnesium absorption in rats

The effects of casein and soybean protein on magnesium absorption and magnesium concentration in the femur were investigated in rats. Purified diets containing either casein or soybean protein and three concentrations of added magnesium (0.82, 1.64 or 2.46 mmol/100 g diet) were used. The isonitrogenous diets were carefully balanced for the different mineral concentrations in the protein preparations. Absolute and percent magnesium absorption and urinary magnesium excretion were significantly decreased in rats fed soybean protein when compared with casein, irrespective of the dietary concentration of added magnesium. The magnesium content of femur was significantly lower in rats fed soybean protein, but this effect was seen only when the diet contained 0.82 mmol magnesium/100 g diet. The addition of sodium phytate to the casein diets, to a concentration identical to that in the diets containing soybean protein as provided by the soybean protein preparation, produced similar effects on magnesium absorption as the diets containing soybean protein. These results indicate that soybean protein, when compared with casein, decreases magnesium absorption through its phytate component.     

Effect of dietary proteins on the plasma immunoreactive insulin-like growth factor-1/somatomedin C concentration in the rat

Plasma immunoreactive insulin-like growth factor-1/somatomedin C (IR-IGF-1) was determined in rats fed for 1 week on a protein-free diet, or diets containing gluten, gluten supplemented with lysine and threonine, maize-gluten meal (with arginine), maize-gluten meal (with arginine) supplemented with tryptophan and lysine, or casein. IR-IGF-1 concentration was higher in the arterial plasma of rats fed on a diet containing casein at 120 g/kg diet (4-8 U/ml) than in rats fed on a protein-free or a low-casein (50 g/kg diet) diet (1.5-2 U/ml). The plasma of rats fed on gluten or maize-gluten meal as the protein source showed intermediate values. However, giving a diet containing an amino acid mixture as recommended by the National Research Council (1978) but deprived of lysine or tryptophan did not affect significantly the plasma IR-IGF-1 concentration. Total IGF-1 concentration (which was measured immunologically after extraction of the plasma with acid-ethanol) was also lower in the rats fed on the protein-free diet than in those fed on the casein (120 g/kg diet) diet. The ratio IR-IGF-1:total IGF-1 was higher in the rats fed on the casein diet (120 g casein/kg diet) than in those fed on the protein-free diet. The results suggest an important influence of IR-IGF-1 or IR-IGF-1:total IGF-1 ratio on protein anabolism and nutrition. IR-IGF-1 and total IGF-1 were found in the fractions of molecular weights 40 kDa and 150 kDa after gel filtration of rat plasma. A larger amount of IGF-1 was recovered in the fraction of 150 kDa in the rats fed on the casein diet. 125I-IGF-1 added to the plasma of rats fed on the protein-free diet was found mainly in the fraction of 40 kDa after gel-filtration. On the other hand, 125I-IGF-1 added to the plasma of rats fed on the gluten or casein diets was mainly recovered in the free IGF-1 fraction. The results suggest that IGF-binding protein(s) of molecular weight about 40 kDa was not saturated with IGF-1 in the rats fed on the protein-free diet. The results indicate the important role of IGF-1 and its binding proteins in the regulation of protein metabolism in rats.     

Bioavailability to rats of iron and zinc in wheat bran: response to low-phytate bran and effect of the phytate/zinc molar ratio

Low-phytate wheat bran was produced by enzymatic hydrolysis and extraction. Rat bioassay methods were utilized to determine bioavailability of iron and zinc in the low-phytate brans and to study the effect of dietary phytate/zinc molar ratio on zinc bioavailability when the phytate source was bran. Endogenous phytase activity hydrolyzed 80-100% of the phytate when wheat bran was incubated in water overnight. The relative biological values of the iron in raw bran and phytate-free bran were 98 and 113, respectively, compared to 100 for ferrous ammonium sulfate in a hemoglobin repletion assay. Low-phytate brans with phytate/zinc molar ratios of 8 or less were equivalent to zinc sulfate as dietary sources of zinc for growth of rats. Rats fed diets that contained wheat bran with zinc sulfate added to reduce the dietary phytate/zinc molar ratio from 40 or 50 to 20 grew at the same rate as rats fed a phytate-free diet, but femur zinc values were lower than those in the reference group. Gel filtration chromatography of extracts of raw and low-phytate brans suggested that zinc might be associated with phytate in wheat bran.