Zinc inhibits nonheme iron bioavailability in humans

There is increasing concern about potential negative interactions in combined iron and zinc supplementation. The aim of the present study was to determine the dose-response effect of zinc, given as a solution, on iron bioavailability. Twenty-two healthy adult women were selected to participate in the study. Iron, with or without zinc was given as an aqueous solution on d 1,2,14, and 15 of the study. Iron bioavailability was measured on the basis of erythrocyte incorporation of⁵⁵Fe or⁵⁹Fe 14 d after administration. Subjects received 0.5 mg of iron together with graded zinc concentrations (0-11.71 mg). No significant effect of zinc on iron absorption was found at Zn : Fe molar ratios up to 2 :1. At 5:1,10:1, and 20 :1 molar ratios, a dose-dependent inhibitory effect on iron absorption was observed (28-40% of iron absorption inhibition; one-way repeated-measures ANOVA, F = 4.48,p = 0.02). In conclusion, zinc administration combined with iron in an aqueous solution leads to the inhibition of iron bioavailability, which occurs in a dose-dependent way. This negative interaction should be considered for supplementation programs with both microminerals.     

Effects of type of fat in the diet on iron bioavailability assessed in suckling and weanling rats

Differences in iron bioavailability from human milk and milk formulas may in part be due to differences in lipid composition. We investigated the short and long term effects of diets based on different fats [corn, coconut, olive, or soy oil, human milk fat (HMF) and a formula fat blend (FF)] on iron absorption in rats. Suckling rat pups dosed with 59Fe-labeled diets containing different fat sources were killed after 6 h, and blood and individual tissues were counted. Iron availability was estimated by % 59Fe in blood. Pups dosed with a more saturated fat (coconut oil) had a higher % 59Fe in blood than those fed other fat sources. Weanling rats were used to determine iron bioavailability from fat sources using both the hemoglobin repletion method and whole body counting. Hemoglobin regeneration was significantly higher for rats fed the HMF diet (8.4 +/- 0.5 g/dl) than from the FF diet (6.5+/-0.6 g/dl) or the corn oil diet (less saturated) (6.4 +/- 0.3 g/dl). Rats fed diets based on coconut oil (more saturated) had significantly higher % 59Fe retention (61.6 +/- 1.4) than rats fed diets based on FF (49.8 +/- 3.4). There was a significant positive association between oleic acid in the diet and oleic acid in the intestinal mucosa (r = 0.95, p < 0.05) and between linoleic acid in the diet and linoleic acid in the intestinal mucosa (r = 0.97, p < 0.05) suggesting that the dietary treatment altered the fatty acid composition of the brush border membrane. Our results suggest that saturated fats may increase iron absorption and that part of this may be achieved by changes in the fatty acid composition of the intestinal mucosa. Hemoglobin regeneration and % 59Fe retention data suggest that differences in iron absorption from infant diets may in part be due to differences in fat composition. Therefore, lipid composition of infant formulas should also be taken into consideration as a factor influencing iron bioavailability.     

Selenium supplementation of infant formula: uptake and retention of various forms of selenium in suckling rats

Formula-fed infants often have lower serum selenium levels than breast-fed infants. Although no deleterious effects have been correlated to this finding, supplementation of formula with selenium is considered. In this study, we investigated the uptake and retention by suckling rat pups of ⁷⁵Se from selenite, selenate, and selenomethionine added to infant formula. The molecular distribution of ⁷⁵Se in liver, kidney, intestine, and plasma was followed by gel-filtration chromatography on Superose 12. ⁷⁵Se-uptake was most rapid from selenomethionine (70% at 1 hr), followed by selenate (51%) and selenite (29%). This difference was explained by a higher retention of ⁷⁵Se in the stomach and small intestinal wall of pups given selenite supplement. Plasma distribution of ⁷⁵Se as studied by gel filtration was also different, with a higher proportion of ⁷⁵Se from selenomethionine being protein-bound than from selenite or selenate. Similarly, a larger proportion of ⁷⁵Se from selenomethionine became protein-bound in the liver than from selenite or selenate. In conclusion, although whole body retention after 24–48 hr was similar, the metabolic fate of selenium varies considerably with the form of selenium added to formula. Further studies are needed to study the long-term consequences of selenium accumulated in different body compartments.     

Zinc and manganese bioavailability from human milk and infant formula used for very low birthweight infants,evaluated in a rat pup model

The bioavailability of zinc and manganese from diets used for very low birthweight infants was investigated in a rat pup model using radioisotopes. The effect of protein source and content and of pasteurization was evaluated, and two different approaches for evaluation of zinc and manganese bioavailability in the rat pup model were compared. Zinc and manganese bioavailability from the studied human milk and infant formula for very low birthweight infants was high. Liver uptake of⁶⁵Zn from labeled premature infant diets in sucklings rat pups was 26–29%, and absorption calculated as the difference between administered dose and nonabsorbed activity 6 h after oral intubation was 93–95%. Retention of manganese calculated as the sum of⁵⁴Mn retained by organs and carcass was 85–95% from human milk and premature infant formula and absorption calculated from nonabsorbed activity was 83–88% after 6 h. Fortification of early human milk significantly increased the bioavailability of zinc. No effect of pasteurization of human milk was found on zinc or manganese bioavailability. Liver zinc uptake was found to be a more sensitive parameter than absorption for evaluation of diets with a high zinc bioavailability. Measurement of retained activity of manganese in carcass and organs was judged to be the preferred parameter for evaluation of diets with high manganese availability.     

Uptake and retention in suckling rats of51chromium fed with human milk or infant formulas

Optimum concentration of Cr for infant formulas has not been established. Such components as soy protein or supplemental Fe could influence absorption and retention. Suckling rat pups were used to evaluate the influence of three commercial formulas and human milk, all of which had been incubated with⁵¹CrCl₃ for 1 h, on the uptake and retention of the added⁵¹Cr. After fasting 3 h, the pups were intubated with a single dose of 25 μCi⁵¹CrCl₃ in either a cow's milk-based formula, an Fe-supplemented cow's milk-based formula, a soy-based formula, or human milk. Six hours later,⁵¹Cr was counted in five organs, thymus, blood, and total urine. Absorption of⁵¹Cr was low. At 6 h, percent⁵¹Cr in blood was <0.2% of the dose, and total⁵¹Cr excretion in urine was <1.8%. The uptake and retention of⁵¹Cr and its concentration in any of the organs, thymus, blood, and urine were not influenced by different types of formula or by human milk.     

l-Lysine supplementation does not affect the bioavailability of copper or iron in rats

l-lysine (Lys) is an essential amino acid that is added to foods and dietary supplements. Lys may interact with mineral nutrients and affect their metabolism. This study examined the effect of dietary Lys supplementation on the bioavailability of copper (Cu) and iron (Fe). Weanling male Sprague-Dawley rats were fed one of five diets (20% casein) for 4 weeks containing normal Cu and Fe (control) or low Cu or Fe without (LCu, LFe) or with (LCu + Lys, LFe + Lys) addition of 1.5% Lys. Final body weights, body weight gains and food consumption of the rats did not differ (P ≥ 0.05) among diet groups. Rats fed the low Cu or Fe diets showed changes in nutritional biomarkers compared to control rats, demonstrating reduced Cu and Fe status, respectively. Hematological parameters, serum ceruloplasmin activity and Cu and Fe concentrations in serum, liver, kidney and intestinal mucosa were unaffected (P ≥ 0.05) by Lys supplementation. These results indicate that in the context of an adequate protein diet, Lys supplementation at a relatively high level does not affect Cu or Fe bioavailability in rats.     

Is solubility in vitro a reliable predictor of iron bioavailability?

In this paper we have described physiological factors that are likely to influence iron bioavailability, reviewed chemical properties of iron important to its absorption, and addressed the question posed in the title. Intestinal transit times, luminal pH, and diffusion barriers resulting from gastrointestinal mucus may affect absorption. The tendency of iron to form large-molecular-weight species and to bind to macromolecules may limit its absorption. Iron solubility following in vitro gastrointestinal digestion is a reliable predictor of ascorbic acid effects on bioavailability but not of protein effects. Solubility of low-molecular-weight iron is better than simple solubility for predicting iron bioavailability.     

In vivo digestomics of milk proteins in human milk and infant formula using a suckling rat pup model

Human milk is the optimal mode of infant feeding for the first several months of life, and infant formulas serve as an alternative when breast-feeding is not possible. Milk proteins have a balanced amino acid composition and some of them provide beneficial bioactivities in their intact forms. They also encrypt a variety of bioactive peptides, possibly contributing to infant health and growth. However, there is limited knowledge of how milk proteins are digested in the gastrointestinal tract and bioactive peptides are released in infants. A peptidomic analysis was conducted to identify peptides released from milk proteins in human milk and infant formula, using a suckling rat pup model. Among the major milk proteins targeted, α-lactalbumin and β-casein in human milk, and β-lactoglobulin and β-casein in infant formula were the main sources of peptides, and these peptides covered large parts of the parental proteins’ sequences. Release of peptides was concentrated to specific regions, such as residues 70–92 of β-casein in human milk, residues 39–55 of β-lactoglobulin in infant formula, and residues 57–96 and 145–161 of β-CN in infant formula, where resistance to gastrointestinal digestion was suggested. In the context of bioactive peptides, release of fragments containing known bioactive peptides was confirmed, such as β-CN-derived opioid and antihypertensive peptides. It is therefore likely that these fragments are of biological significance in neonatal health and development.     

Intestinal absorption of copper from drinking water containing fulvic acids and an infant formula mixture studied in a suckling rat model

The purpose of this study was to investigate if the intestinal absorption of copper in drinking water is altered in the presence of complexing agents from a fulvic acid mixture and an infant formula powder. Ten to twelve day old rat pups were given a single oral dose of radio-labeled Cu in deionized water (0.93 mg Cu/l), in water containing fulvic acids (10 mg/l), in infant formula mixed with deionized water, or in infant formula mixed with water containing fulvic acids. Six hours after dosage, radioactive Cu was analyzed in the mucosa of the small intestine, the liver and the remaining carcass (excluding the liver and gastrointestinal tract) by gamma counting. Dialysis and centrifugation experiments showed that Cu was complexed by components in the fulvic acid and formula mixtures, although the presence of fulvic acids in the water did not alter the Cu fractionation in the formula. The fractional Cu uptake (% of dose) from the intestinal lumen to the mucosa was not markedly changed by the presence of the chelating agents. However, the retention of Cu in the intestinal mucosa was increased by both fulvic acids and formula. Concomitantly, the absorption rate of Cd to the circulatory system was decreased. No interactive effect between fulvic acids and formula was found on the Cu absorption. These findings indicate that the water quality may be an important determinant of the rate of intestinal Cu absorption from drinking water. Moreover, in the future risk assessment of copper in drinking water, the possibility of alterations in absorption of drinking-water Cu has to be considered when the drinking water is used for cooking.     

Humoral and cellular immune responses in gluten-treated suckling or hand-fed rats

We analyzed the immune response to gliadin in suckling rats and rats hand-fed with an artificial milk formula, an animal model of gluten enteropathy. Animals of both groups were intragastrically given either gliadin or albumin (control animals) or gliadin from birth till day 55. When compared to the controls, spleen lymphocytes from both groups of gliadin-treated rats cultivated in vitro exhibited a significant increase of spontaneous 3H-thymidine incorporation. Moreover, the proliferation of spleen and mesenteric lymph node (MLN) lymhocytes from both groups of gliadin-treated suckling and hand-fed rats was specifically increased by the in vitro gliadin challenge. Spleen B cells from gliadin-treated rats spontaneously produced higher amounts of gliadin-specific antibodies than those from the controls, however, in vitro stimulation by gliadin caused no further increase in antibody production. Apoptotic DNA fragmentation in MLN cells was higher in gliadin-treated rats than in albumin-treated ones, independently of the milk diet during the suckling period.     

Zinc uptake by rice as affected by iron and a chelator of ferrous iron

Summary Iron competitively inhibited Zn absorption by rice (Oryza sativa L. cv. Earlirose) grown in solution culture. The effect was more marked for shoots since Fe had also a competitive effect on Zn translocation from roots to shoots. The chelating agent baptholphenanthrolinesulfonate (BPDS), which has great ability to chelate Fe⁺⁺, alleviated the inhibitory effect of Fe to a large extent. re]19750516     

Lack of stimulation of relaxin secretion in lactating sows by suckling in vivo or by oxytocin in vitro.

After parturition, eight sows were zero weaned by removing all piglets 6 h after birth; a further 18 sows suckled at least ten piglets each. Blood samples were collected on Day 4 after zero weaning or on Days 4, 14 and 21 of lactation and the sampling frequency increased during suckling bouts. Ovaries were recovered from sows on these days and corpora lutea were either extracted for estimation of relaxin and progesterone concentration, fixed for immunohistochemical analysis or incubated in vitro in the presence or absence of luteinizing hormone (LH) or oxytocin. Luteal weight and progesterone were higher in the zero-weaned sows than in lactating sows (P less than 0.05 and less than 0.001, respectively); relaxin content was below detection by Day 14. This was supported by immunohistochemical staining for relaxin, which showed limited immunostaining in zero-weaned and Day 4 sows, but none in the tissue recovered on Days 14 and 21, which showed typical signs of regression. Secretion of progesterone and relaxin by luteal tissue in vitro was highest in zero-weaned sows (P less than 0.05), decreased as lactation progressed and neither LH nor oxytocin had any significant effect. Concentrations of plasma relaxin were all less than 0.2 ng/ml in three of the four zero-weaned and Day-4-suckled sows assayed; there was no detectable increase during suckling bouts. It was concluded that during lactation the old corpus luteum of pregnancy is not able to release relaxin in response to suckling in vivo or to oxytocin treatment in vitro.     

Enhanced lutein bioavailability by lyso-phosphatidylcholine in rats

The bioavailability of lutein solubilized in mixed micelles containing either phosphatidylcholine (PC) or lysophosphatidylcholine (lysoPC) was evaluated in male rats. Mixed micelles contained 2.5 mM monooleoylglycerol, 7.5 mM oleic acid, 12 mM sodium taurocholate and 200 μM lutein either with 3 mM PC or lysoPC. To study lutein bioavailability, single and repeated dose experiments were conducted. For single dose study, group of rats (n = 30/group) were fed single dose of lutein solubilized in lysoPC (LPC group), PC (PC group) and no phospholipids (NoPL group) in micellar form. Each group was further divided in to five sub-groups (n = 6/sub group) to measure lutein bioavailability over time up to 9 h. For repeated dose study, group of rats (n = 6/group) were fed daily for 10 days a dose of lutein in mixed micelles with NoPL, PC and LPC. A separate group (n = 6) not fed mixed micelles was considered as zero-time control. In both the experiments, mixed micelles (0.2 ml/rat) were fed to the rat by direct intubation to the stomach. Results of single dose studies showed that the mean lutein levels in the plasma and liver of the PC group was significantly lower (p < 0.05) than those of the other two groups. Moreover, the average lutein level in the plasma and liver was significantly (p < 0.05) different among the groups in the order LPC > NoPL > PC. But, repeated dose experiment followed the order LPC > PC > NoPL. The level of lutein excreted through urine and feces of PC group was significantly higher (p < 0.05) than those of the other two groups. Thus, the results indicate that the PC in the mixed micelles suppressed the intestinal uptake of lutein after single dose but not after repeated dose and that lysoPC enhanced the absorption. In both the experiments, plasma and liver level of lutein was higher in LPC compared with PC group. Results also suggest that the luminal hydrolysis of PC to lysoPC is necessary for intestinal uptake of lutein solubilized in mixed micelles.     

Long-term effects of iron: Zinc interactions on growth in rats

The influence of iron (Fe) on the bioavailability and functional status of zinc (Zn) was studied in young rats using metabolic balances and tissue dosages, which were compared to growth. Diets supplied adequate intakes of Fe (45 and 300 mg/kg diet) and Zn (14 and 45 mg/kg) for 2 mo. Two metabolic balance determinations were performed that were correlated for Zn and Fe during the first and the last weeks of the study. A significant effect of Fe supply, but not of Zn was displayed on Fe absorption; both Fe and Zn diet concentrations had a significant influence on Zn absorption. Fe and Zn organ contents were significantly correlated with the amount absorbed during the two metabolic balances. There was a positive correlation between liver and muscle Fe and Fe absorption, and Fe absorption and muscle Zn, as well as a negative one with liver Zn; a positive correlation was displayed between Zn absorption and Zn organ content. No correlation was found between Zn absorption and Fe tissue content. Growth was correlated with Zn, but not with Fe absorption during both balances. A positive correlation was displayed between growth and Zn liver content, and a negative one with Fe liver content. Care must be taken to give growing subjects balanced diets or supplementation, since the negative interactions between these trace elements are likely to persist as long as the diet is given.     

A case of the adoption of an infant chimpanzee by a suckling foster chimpanzee

Chimpanzees in captivity have grown up in a rather unnatural social environment and there frequently are problems when they have to nurse their own offspring. It is most remarkable that a chimpanzee mother in a captive colony, who had lost her child almost immediately after birth, adopted without problems a five-week-old infant, which had been reared by humans from the day of its birth. Successful adoption has not been reported for feral chimpanzees; similar cases in captivity are not known.     

Signs of iron deficiency in copper-deficient rats are not affected by iron supplements administered by diet or by injection

The goal of this study was to determine the effects of Fe supplementation on the anemia of Cu deficiency in rats. In addition, we observed changes in serum and organ Cu and Fe during the development of Cu deficiency. In Experiment 1, weanling male Sprague-Dawley rats were fed AIN-93G diets containing either <0.3 mg Cu [Cu deficient (CuD)] or 6.0 mg Cu [Cu adequate (CuA)] per kilogram diet, and 35 mg Fe/kg. Five rats from each group were killed at intervals for the analysis of hematologic parameters and mineral content of various organs. In Experiment 2, two groups of 24 rats each were fed either the CuA diet or the CuD diet for 14 days. Then, three sets of eight rats in each group received three separate Fe treatments: (1) daily intraperitoneal injections of 400 mug Fe (Cu-free ferric citrate) per rat for another 14 days, (2) fed similar diets that contained three times the normal amount of Fe (105 mg/kg) for 14 days, or (3) received no further Fe treatment. At day 21, all rats were fed a 1-g meal labeled with (59)Fe to determine Fe absorption. After 28 days, rats were killed for the analyses of Fe and Cu status. Results of Experiment 1 showed that within 14 days, CuD rats had lower blood hemoglobin (Hgb), red blood cell count, and mean corpuscular volume than CuA rats. Copper concentrations in all tissues measured were lower in the CuD rats than in controls. Serum ceruloplasmin (Cp) activity in CuD rats was only 0.8% of CuA rats at day 7. During this period, enterocyte and liver Fe concentrations were elevated and serum Fe was reduced, but there was no change in spleen Fe. Results of Experiment 2 showed that CuD rats absorbed less Fe than CuA rats. Supplemental Fe by diet or by intraperitoneal injections did not prevent anemia in the CuD rats or affect other parameters of Cu status. Serum total iron binding capacity [transferrin (Tf)] was not changed by Cu deficiency or by Fe supplementation; however, percent Tf saturation was reduced in CuD rats but was not enhanced by Fe supplementation. These data suggest that anemia of Cu deficiency occurs because of reduced Fe absorption, and it inhibits release of Fe from the liver and inefficient loading of Fe into Tf because of very low plasma Cp activity. The latter then leads to inefficient delivery of Fe to the erythroid cells for heme and Hgb synthesis.     

Potential role of in vitro iron bioavailability studies in combatting iron deficiency: a study of the effects of phosvitin on iron mobilization from pinto beans

Iron deficiency and iron overload affect one billion people worldwide. Treatment of iron malnutrition can be enhanced by an understanding of iron bioavailability from the diet. We have focused on the development of in vitro methods for determining iron bioavailability in the hopes of providing both an understanding of the chemical basis leading to the inhibition or enhancement of iron absorption and the provision of methodologies which will allow nutritionists around the world to ascertain iron bioavailability of local foods and food combinations. The study reported here focuses on the effects of phosvitin, a suspected inhibitor of iron absorption found in egg yolks, on the chemistry of iron during the in vitro enzymatic digestion of pinto beans. Three basic types of information were obtained. First, the total soluble iron was determined during in vitro enzymatic digestion under simulated oral, gastric (pH 2) and duodenal (pH 6) conditions. Phosvitin was found to have a strong solubilizing effect at pH 6 and pH 2 when in the presence of ascorbate. Pyrophosphate also leads to high iron mobilization. A second approach is to determine the static Fe2+ and Fe3+ concentrations following in vitro enzymatic digestion of pinto beans at pH 2 and pH 6. Ascorbic acid enhanced the total soluble iron at both pH values, however, only at pH 2 was a large proportion of the iron found in the Fe2+ state and then only in the presence of phosvitin but not pyrophosphate. A third approach is to determine the amount of Fe2+ formed in the digestive supernatant during a 10-min incubation with ferrozine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Intestinal transport of manganese from human milk, bovine milk and infant formula in rats

The transport of manganese from extrinsically labeled human milk, bovine milk and infant formula was studied by the everted intestinal sac method. Tissue/mucosal flux data indicated that transport of manganese into the intestinal tissue was significantly greater with bovine milk and formula than from human milk. Similarly, the total flux of manganese from the mucosal to serosal surface was less when human milk was used. Smaller molecular weight manganese binding ligands isolated from the milk samples enhanced the mucosal to tissue movement of manganese as contrasted to the higher molecular weight manganese binding ligands. Most significantly the data suggest that the transport and uptake of manganese is less in the presence of human milk and its isolated manganese fractions than it is in bovine milk or infant formula.     

Amino acids and lactate efflux from in vitro incubated soleus muscles of suckling rats.

The net in vitro release of lactate, pyruvate, alanine, glutamate, glutamine and total amino acids from soleus muscles of suckling rats was studied. In all cases, the net output of metabolites was considerably higher in young animals than in 30 day old controls. Glutamine efflux of 1 day old pups was very low, probably due to lack of effectivity of the glutamine synthesizing system. The considerable output of amino acids in the younger animals was partly due to depletion of internal amino acid pools.