Copper absorption from foods labelled intrinsically and extrinsically with Cu-65 stable isotope.

OBJECTIVE: To determine copper absorption from copper containing foods labelled either intrinsically or extrinsically with a highly enriched Cu-65 stable isotope label. DESIGN: A longitudinal cross-over study. SETTING: The study was conducted at the Institute of Food Research, Human Nutrition Unit, Norwich, UK. SUBJECTS: Subjects were recruited locally via advertisements placed around the Norwich Research Park. A total of 10 volunteers (nine female, one male) took part in the study, but not all volunteers completed each of the test meals. INTERVENTIONS: A highly enriched Cu-65 stable isotope label was administered to volunteers in the form of a reference dose or in breakfast test meals consisting of red wine, soya beans, mushrooms or sunflower seeds. Faecal monitoring and mass spectrometry techniques were used to estimate the relative quantities of copper absorbed from the different test meals. RESULTS: True copper absorption from the reference dose (54%) was similar to extrinsically labelled red wine (49%) and intrinsically labelled sunflower seeds (52%), but significantly higher than extrinsically labelled mushrooms (35%), intrinsically (29%) and extrinsically (15%) labelled soya beans and extrinsically labelled sunflower seed (32%) test meals. CONCLUSIONS: The use of Cu-65 extrinsic labels in copper absorption studies requires validation according to the food being examined; intrinsic and extrinsic labelling produced significantly different results for sunflower seeds.

Chronic exposure to high levels of dietary iron fortification increases lipid peroxidation in the mucosa of the rat large intestine.

There is increasing evidence that excess dietary iron may be a risk factor for colorectal cancer. However, the majority of animal studies looking at possible mechanism have used unrealistically high concentrations of iron. The current study was designed to test whether chronic exposure to high levels of iron fortification affects the free radical generating capacity of the lumenal contents, mucosal lipid peroxidation and crypt cell proliferation. Rats were fed diets containing either 29 mg/kg or 102 mg/kg of elemental iron for 6 mo. The free radical generating capacity of lumenal contents was assessed using an in vitro assay. Crypt cell proliferation rate was measured in tissues taken from the cecum and colon, with the remaining tissue being used for the assessment of lipid peroxidation. Chronic feeding of iron did not increase crypt cell proliferation rate in either the colon or cecum, but it was associated with an increase in free radical generating capacity in the colon and increased lipid peroxidation, particularly in the cecum. These results may be relevant to epidemiological evidence showing that dietary iron is associated with the risk of proximal colon cancer in humans.

Oral ferrous sulfate supplements increase the free radical-generating capacity of feces from healthy volunteers.

BACKGROUND: Most dietary iron remains unabsorbed and hence may be available to participate in Fenton-driven free radical generation in conjunction with the colonic microflora, leading to the production of carcinogens or direct damage to colonocytes. OBJECTIVE: Our aims were to measure the proportion of fecal iron available to participate in free radical generation and to determine the effect of an oral supplement of ferrous sulfate on free radical generation. DESIGN: Eighteen healthy volunteers recorded their food intake and collected fecal samples before, during, and after 2 wk of supplementation (19 mg elemental Fe/d). Total, free, and weakly chelated fecal iron were measured and free radical production was determined by using an in vitro assay with dimethyl sulfoxide as a free radical trap. RESULTS: Fecal iron increased significantly during the period of supplementation and returned to baseline within 2 wk. The concentration of weakly bound iron in feces (approximately 1.3% of total fecal iron) increased from 60 micromol/L before to 300 micromol/L during supplementation, and the production of free radicals increased significantly (approximately 40%). Higher-carbohydrate diets were associated with reduced free radical generation. CONCLUSION: Unabsorbed dietary iron may increase free radical production in the colon to a level that could cause mucosal cell damage or increased production of carcinogens.

Increases in the concentrations of available iron in response to dietary iron supplementation are associated with changes in crypt cell proliferation in rat large intestine.

High concentrations of iron in the diet have been shown to increase chemically induced colorectal tumors in rats. It is therefore important to understand the influence of dietary iron on the concentration of unabsorbed iron in the large intestine and its distribution between soluble and insoluble pools in the luminal compartment. We sought to investigate this issue and to establish whether iron modifies mucosal cell proliferation, which is thought to influence initiation and progression through the adenoma carcinoma sequence. In the first experiment, four groups of seven rats were fed diets at two concentrations of iron, 29 and 102 mg/kg, with or without the addition of 2.5 g phytic acid/kg. The concentrations of iron in the contents of the large bowel extractable with water ("free iron") or a buffered EDTA solution ("exchangeable iron") were determined. The concentration of freely soluble iron increased approximately 100% with iron supplementation in both the cecum and the colon, and there was an approximately five- to sixfold increase in exchangeable iron at both sites (P < 0. 05). In a second experiment with identical feeding conditions, there was a significantly greater number of cell divisions per crypt in the colon of the high iron group and a significantly greater number of cell divisions in the upper part of the crypt in the cecum. The concentrations of free and exchangeable iron observed in colonic contents in this study are consistent with those reported by others to increase free radical production in fecal material. Further studies are required to determine whether the small changes in crypt cytokinetics are a consequence of oxidative mucosal damage.

Factors affecting iron stores in infants 4-18 months of age.

OBJECTIVES: To determine the effects of dietary, physiological or environmental factors on body iron levels in infants aged 4-18 months. DESIGN: The daily iron intake of the infants was measured from a diet history obtained by interview using a standardised question sheet, previously validated against weighed intake (minimum 3 days) in an independent sample of 8 and 18 month old infants. Capillary blood samples were analyzed for haemoglobin, mean cell volume, haematocrit, zinc protoporphyrin and plasma ferritin concentration. Ferritin values were log-transformed prior to analysis to give a better approximation to the normal distribution and forward stepwise multiple linear regression was carried out using SPSS. SETTING: The city of Norwich, UK and some of its suburbs. SUBJECTS: One hundred and eighty-one healthy infants in age groups 4, 8, 12 and 18 months. RESULTS: Main determinants of iron stores in the 4 month old infants were birth weight (+ve (P < 0.001)) and body weight (-ve (P < 0.005)). In the 8 month old infants intake of cow's milk (-ve (P < 0.05)), belonging to a smoking household (-ve (P < 0.05)) and quantity of commercial babyfood consumed (+ve (P < 0.05)) were significant. In this age group there was a gender effect (girls > boys (P < 0.01)) and the gender effect remained at 12 months (girls > boys (P < 0.05)), but at 18 months only non-haem iron intake was a significant factor (-ve (P < 0.05)). CONCLUSIONS: At 4 months of age birth weight and body weight exert the greatest influence on iron stores, whereas by 8 months components of the weaning diet have an effect (commercial babyfood (+ve), cow's milk (-ve)); there is also a gender effect (girls > boys), possibly reflecting the different growth rate between boys and girls. At 12 and 18 months the only significant factors are gender (girls > boys) and non-haem iron intake (-ve) respectively.

Effect of calcium supplements and stage of lactation on the calcium absorption efficiency of lactating women accustomed to low calcium intakes.

The effect of calcium intake on the calcium absorption efficiency from 100 mL cow milk was measured in lactating Gambian mothers habituated to a low-calcium diet [mean intake 7.08 mmol (283 mg)/d], and compared with UK lactating mothers consuming high-calcium diets [mean intake 29.2 mmol (1168 mg)/d] by using a double stable-isotope technique (oral 44Ca and intravenous 42Ca). In a double-blind trial starting 9 d postpartum, Gambian mothers were given a calcium supplement [17.85 mmol (714 mg)/d] or placebo for 12 mo. At 3 and 12 mo postpartum, mean (+/- SEM) calcium absorption from isotopically enriched milk was 52.3 +/- 3.1% (n = 25) and 47.2 +/- 4.8% (n = 24) in the unsupplemented Gambian mothers and 48.8 +/- 2.8% (n = 28) and 42.9 +/- 3.7% (n = 24) in the supplemented mothers, respectively. There was no effect of supplementation or stage of lactation on the efficiency of calcium absorption. At 3 mo postpartum the UK mothers absorbed 32.2 +/- 3.8% of the isotopically enriched calcium added to milk, which was significantly less than that of the Gambian mothers (P < 0.01).

Zinc absorption in infants fed iron-fortified weaning food.

The effect of fortification iron (reduced iron) on zinc absorption from a commercial vegetable-based weaning food was assessed in 11 9-mo-old infants. Each infant was fed a test meal of unfortified or iron-fortified product, labeled extrinsically with 1 mg 67Zn or 70Zn (as citrate), and the next day was fed the second product labeled with the other isotope. A complete fecal collection was carried out for 3-4 d, and the amount of unabsorbed isotope measured by thermal-ionization quadrupole mass spectrometry. Apparent zinc absorption (isotope intake minus fecal excretion, expressed as the % of dose administered) was 31.1 +/- 8.3% (x +/- SD) from the iron-fortified food and 28.6%28.6 +/- 10.5% from the unfortified food. These values were not significantly different, thus iron fortification of the weaning food did not reduce zinc absorption.

The bioavailability of iron in different weaning foods and the enhancing effect of a fruit drink containing ascorbic acid.

There is limited information on the bioavailability of Fe in infant weaning foods, mainly because of the difficulties of measuring Fe utilization directly in infants. The aim of this study was to develop a safe and relatively noninvasive method for studying Fe bioavailability (measured as percent Fe incorporation into red blood cells) in infants using 54Fe, 57Fe, and 58Fe stable isotopes. Four commonly used weaning foods were selected for study, labeled extrinsically with 57Fe- or 58Fe-enriched ferrous sulfate, and fed to five female and five male 9-mo-old fasting infants, using a multiple-dosing technique. Each food was given three times, labeled with one isotope, with a fruit juice drink containing 50 mg of ascorbic acid, and three times, labeled with a different isotope, with an ascorbic acid-free drink. Fourteen days after the last test meal, a blood sample was obtained from a heel-prick, spiked with a known amount of 54Fe, digested, and purified by ion exchange; isotopic enrichment and total Fe content were measured by quadrupole thermal ionization mass spectrometry. The proportion of administered dose of isotope circulating in the blood was calculated from an estimate of blood volume. The geometric mean bioavailability (range) was 3.0% (1.2-9.5%) in a proprietary dehydrated vegetable product, 3.0% (1.1-21.2%) in Weetabix whole-wheat breakfast cereal, 3.1% (1.2-15.4%) in wholemeal bread, and 4.3% (1.7-10.3%) in baked beans. When taken with the drink containing ascorbic acid, there was a 2-fold increase in bioavailability in all foods except the vegetable meal, presumably because this was already fortified with ascorbic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Assessment of zinc bioavailability: studies in rats on zinc absorption from wheat using radio- and stable isotopes.

Absorption from wheat intrinsically and extrinsically labelled with 67Zn and extrinsically labelled with 65Zn was measured from 67Zn faecal excretion and 65Zn whole-body retention in rats. There were significant differences between the extrinsically- and intrinsically-labelled 67Zn (P < 0.001), but not between the extrinsically-labelled 65Zn and intrinsically-labelled 67Zn. The effect of chicken meat on the absorption of Zn from intrinsically-labelled wheat was also studied in the rat. Mean Zn absorption from wheat and chicken meat fed separately was 18.5 and 68.2% respectively, and from a mixture of the two containing the same level of Zn was 50.1%. The apparent absorption of Zn from the composite meal was significantly higher than predicted from the results of the foods on their own (P < 0.001).

The measurement of exchangeable pools of zinc using the stable isotope 70Zn.

The present study was designed to assess the feasibility of using small doses of a stable isotope of Zn to follow plasma kinetics over a 10 d period and, hence, make deductions about Zn turnover and body pool sizes. At the beginning of the 10 d metabolic balance, two adults, consuming their habitual diet, were given an intravenous injection of 70Zn. There was a fourfold difference in the administered dose between the two subjects (0.445 and 2.078 mg). Blood samples were taken at regular intervals and plasma enrichment with 70Zn measured by thermal ionization mass spectrometry. Urine and faeces were collected and analysed for Zn and 70Zn. Kinetic analysis of the plasma 70Zn decay by several different methods was undertaken. It was apparent from both deconvolution analysis of the short-term (0-90 min) decay data and four-compartment modelling of the longer-term (0-24 h) data that isotopic Zn very rapidly equilibrates with the plasma Zn and with a rapidly exchanging non-plasma pool, probably located within the liver. This latter pool appears to contain less than 10 mg Zn and the peak of isotope enrichment occurs at about 20 min post injection. The later decay of plasma Zn enrichment appears to be dictated by exchange with a much larger pool of approximate size 350 mg.

Zinc absorption in adult men from a chicken sandwich made with white or wholemeal bread, measured by a double-label stable-isotope technique.

Eleven fasted adult men consumed a chicken meat sandwich made with white or wholemeal bread, extrinsically labelled with 2 mg 67Zn, on two different occasions. Immediately after eating the sandwich they were given an intravenous injection of 1.5 mg 70Zn. True Zn absorption (which was approximately 7% higher than apparent absorption) was determined by the faecal balance technique by making an allowance for endogenous excretion from measurements of faecal excretion of 70Zn. There was no significant difference in mean true Zn absorption from the white or wholemeal bread sandwich, 33.6 and 25.4% respectively. It was concluded that the substitution of wholemeal for white bread does not reduce Zn absorption from meat-based sandwiches.

A preliminary study of the bioavailability of iron- and zinc-glycine chelates.

Groups of rats were fed diets containing marginal levels of Fe and Zn as glycine chelates (tradename 'Chelazome', Albion Laboratories, Verona, New Jersey, USA), or the same level of mineral as ferrous sulphate or zinc carbonate. The Fe diets were fed to weanling rats for 4 weeks and the Zn diets to young adult rats for 5 weeks. Blood Hb concentrations were significantly higher in the group fed Fe-chelazome than ferrous sulphate, 149 and 128 g/l respectively (P less than 0.001), but PCV and liver Fe concentrations were similar between the two groups. No difference in plasma Zn, pancreas, testes or femur Zn concentrations were observed between the two Zn groups, indicating that Zn-chelazome has no advantage over zinc carbonate. The results of this preliminary study indicate that Fe-chelazome has a higher bioavailability than ferrous sulphate and merits further study.

Apparent zinc absorption by rats from foods labelled intrinsically and extrinsically with 67Zn.

A variety of foods (peas (Pisum sativum), chicken meat, eggs, goat's milk, human milk) enriched with the stable isotope 67Zn were prepared by means of intrinsic- and extrinsic-labelling procedures. They were fed to rats and apparent absorption of 67Zn determined from faecal excretion measurements using thermal ionization mass spectrometry. There were significant differences in the absorption of the extrinsic and intrinsic label which differed in magnitude between the foods tested. The extrinsic 67Zn was less well absorbed in peas, chicken meat, eggs, and human milk than intrinsic 67Zn, but in goat's milk the extrinsic 67Zn was better absorbed than the intrinsic label. These results demonstrate that extrinsically-added stable Zn isotopes do not fully exchange with endogenous Zn in many foods, and illustrate the need for caution when using extrinsic labels for Zn bioavailability studies.

Intrinsic labelling of different foods with stable isotope of zinc (67Zn) for use in bioavailability studies.

Intrinsically-labelled foods are required to validate extrinsic-labelling techniques used to study the bioavailability of trace elements. Wheat (Triticum aestivum), peas (Pisum sativum), goat's milk, human milk, eggs and chicken meat were selected for intrinsic-labelling studies with 67Zn. Peas were grown hydroponically in enriched nutrient solution and wheat was grown in sand and watered with enriched nutrient solution. Some of the wheat plants were also given stem injections of 67Zn solution. Eggs and chicken meat were prepared by administering 67Zn intravenously to chickens, and human milk was collected after an oral dose of 67Zn in a cola drink. All the foods investigated were sufficiently enriched with 67Zn for Zn absorption studies except wheat prepared by the sand and water-culture method.

Limited compensation by table salt for reduced salt within a meal.

Sixteen subjects, all of whom had said in a preliminary questionnaire that they normally added table salt to foods, were fed standard meals in the laboratory over 10 days. The meals were identical, except that on 5 days the meal had no added salt (containing 0.46 g sodium chloride) or had salt added to a level of 5.09 g. They were allowed free access to salt pots with the meals and used an average of 1.40 g table salt with the unsalted meal and 0.36 g with the salted meal, thus compensating for 22% of the difference in salt content of the meal. There was no difference in water consumption between the two types of meal. Nutrient intake from the rest of the diet did not differ between periods with high and low salt meals. The failure to compensate more fully for reduced salt in the foods can be attributed to the greater availability of table salt for perception; less table salt than salt incorporated in the foods is therefore required. Reduction of salt concentrations in purchased foods would be unlikely to be fully replaced by the consumer adding table salt.

Validity of calculating fatty acid intake from mixed diets.

One hundred and seventy six weighed duplicate diets were collected over 16 consecutive days from 11 subjects. Analyses of their fatty acid composition were used to asses the validity of food composition tables. Four different calculating techniques were employed. Using the published data produced correlation coefficients of 0.29 between analysed and calculated polyunsaturated/saturated fatty acid ratios, whilst the addition of new analytical data and recoding fried foods produced a correlation coefficient of 0.56. The latter method also decreased the mean difference between analysed and calculated polyunsaturated/saturated consumption, when compared with the standard procedure.

Fatty acid composition of selected foods consumed in a mixed diet study.

In a 16 day dietary survey of 11 subjects, twenty foods were consumed which contributed significantly to the fat intake of the group, but on which fatty acid data were not available. These twenty foods were sampled using the procedure described in the published food composition tables and analysed for their fatty acid composition. The sampling procedure and results of fatty acid analysis are presented.