Studies with anaemic children and pregnant women from areas where vitamin A deficiency is endemic have shown a beneficial effect on Fe status of supplemental vitamin A in addition to Fe supplementation. This suggests a relationship between vitamin Aand Fe status, which we attempted to mimic in rats with anaemia and chronic vitaminA deficiency. Male rats were fed on Fe-adequate diets (35 mg Fe/kg)containing different levels of vitamin A (1200,450,150,75 and 0 retinol equivalents (RE)/kg feed) until they were 5 weeks old. These diets wereidentical to the diets fed to their mothers. Then the young male rats were transferred to diets containing the same levels of vitamin A but no added Fe. After another 2 weeks the rats wererepleted with Fe (35 mg/kg feed) without or with vitamin A to a level of 1200 RE/kg feed. Increased vitamin A intake by the groups previously fed on diets with either 0 or 75 RE/kg produced a reduction in blood haemoglobin concentration, packed cell volume and erythrocyte count. In the group which had been fed on the diet without vitamin A, supplemental vitamin A raised mean cell volume, plasma Fe concentration and total Fe-binding capacity. Vitamin A supplementation during the period of Fe repletion produceda decrease in splenic and tibia Fe concentration, the effect being greater with increasing seventy of previous vitamin A deficiency. The paradoxical effect of supplemental vitamin A on haemoglobin, packed cell volume and erythrocyte count can be explained by a decrease in the degree of haemwoncentration. Thus, the positive effect of supplemental vitamin A seen in humans is also observed with rats under controlled experimental conditions. We speculate that supplemental vitamin A during Fe repletion contributes to optimum erythropoiesis and Fe mobilization when baseline vitamin A status is impaired

Triacylglycerol-rich lipoproteins (TGRLP) were isolated from Cu-deficient and control rats. TGRLP from Cu-deficient rats appeared more fluid than those from controls as sensed by the fluorescence anisotropy of 1,6-diphenyl-l-3,5-hexatriene (DPH). This high fluidity was related to alow cholesterol: phospholipid ratio and high triacylglycerol content in these lipoproteins. TGRLP from Cu-deficient rats were more susceptible to in vitro peroxidation than lipoproteins from control rats as shown by the rate of diene conjugation. The damage induced by the peroxidation resulted in a more ordered state of the lipid fraction especially in lipoproteins from Cu-deficient rats. Thus, after in vitro peroxidation, TGRLP from Cu-deficient rats were more rigid than those from controls. These results suggest that Cu deficiency induces modifications in physicochemical properties of TGRLP which could affect their metabolism.

The hypothesls was tested that fermentable dietary fibre (DF) sources elevate faecal N excretion at the expense of urinary N without affecting N retention. DF that substantially increase fermentation (pectin, Sugarbeet and soya bran) or are poorly fermented (crystalline cellulose and maize bran) were fed as supplements to a basal DF-free diet a t three dose levels: 0,50 and 100 g supplement/kg basal diet. The diets were fed to juvenile male Wistar rats for 2 weeks before a 7 d period when faeces and urine were collected. Faecal excretion of N was significantly increased, dose-dependently, by all DF supplements and was positively correlated to faecal bulking. Urinary excretion of N was lower at the high doses of the DF supplements but reached significance only with the highly fermentable (0·68) sugarbeet- supplementeddiets. Regression analysis showed that the major part (0·75) of the increase in faecal N excretion due to DF supplementation was balanced by a reduction in urinary excretion; N retention was therefore, at the dose levels used, only affected to a small extent. Only in the maize-bran-supplemented diets were the reductions in N retention significant. The shift in N excretion from urine to faeces can be explained largely by the degree of microbial fermentation in the large intestine caused by the addition of DF supplements and emphasizes the modifying role that certain DF supplements may have on the enterohepatic cycle of N. Possible implications of these findings for patients with liver or renal failure or for conditions when the intake of dietary protein is marginal are discussed.

The aims of the present study were to establish a suitable criterion for estimating the vitamin E requirement for young pigs, and to investigate the influence of dietary fats on the requirement of this nutrient. In Expt 1 weaned pigs were given a semi-purified diet supplemented with 0,20, or 100 mg DL- α-tocopheryl acetate/kg. Pigs in Expt 2 were fed on diets containing 10 g sunflower oil (SO) stripped of vitamin E/kg in diets 1 and 2, 100 g SO/kg in diets 3 and 4, and 100 g tallow/kg in diet 5. Diets 2, 4 and 5 were supplemented with DL-α-tocopheryl acetate at 20 mg/kg for 0·28 d and 50 mg/kg for 29–56 d of the experiment respectively. Results showed that vitamin E concentrations in plasma and tissues reflected dietary levels of vitamin E. No apparent clinical signs of vitamin 'E deficiency were observed, but pigs fed on diets without vitamin E supplementation showed a higher (P < 0·05) lipid peroxidation status as indicated by thiobarbituric acid-reactive substances in erythrocytes, and ethane and pentane levels in exhaled gases than those fed on supplemental diets. This indicated that the former was deficient in vitamin E. Data in Expt 1 suggested that supplementation with 20 mg DL-α-tocopheryl acetate/kg is adequate for young pigs when the diet contains 30 g lard/kg. However, results in Expt 2 showed that this level was inadequate when the diet contains 100 g SO/kg or its equivalent to 70 mg polyunsaturated fatty acids (PUFA)/g. Vitamin E and lipid peroxidation status of pigs were affected by both dietary vitamin E and dietary PUFA. This demonstrates that the requirement for vitamin E in young pigs increases as PUFA levels in the diet increase. The present study shows that lipid peroxidation response of pigs is a suitable index to evaluate vitamin E requirements.

Several studies have failed to demonstrate an association between Fe status and intake of dietary Fe. However, in the long term, it Seems logical to presume that body Fe reserves are, fundamentally, dependent on the intake of bioavailpble dietary Fe. This discrepancy may depend on several factors(1)interindividual variation in biological availability of dietary Fe (differences in intestinal absorption),(2) interactions between dietary Fe and absorption enhancers and inbibitors, (3) variatiom in physiological (menstruation, childbirth) or unphysiological (blood donation) Fe losses, (4) the failure to adjust dietary intake data for Fe supplements, (5) uncertain food composition data (discrepancies between calculated and chemically measured Fe content inthe diet), and (6) diet reporting error (reported intake of dietary Fe may deviate considerably from the true intake). The present study examined associations between dietary intake of Fe (assessed by diet history interview) and Fe status (assessed by femtin status) among 167 Danish women aged 35–65 years, who were not blood donors, by taking into account diet reporting error (assessed from p-amino benzoic acid-validated urinary N), physiological blood losses (menstruation, Childbirth, abortions), and Fe supplementation. Our results indicate that the lack of a general association between Fe status and dietary Fe intake may, in part, be caused by selective dietreporting error.

Fractional apparent absorption of Zn and Ca from a wheat-milk-based infant cereal was studied in six healthy infants (18-30 weeks old). Mineral absorption was measured by a stable-isotope technique based on faecal excretion of the isotopes. Each test meal (40 g cereal) was extrinsically labelled with 70Zn and 42Ca before intake. All faecal material passed during the 21 d following intake of the labelled test meal was collected on trace-element-free nappies. Individual stool samples were analysed for their content of ‘OZn and 42Ca by thermal ionization mass spectrometry. Apparent absorption was calculated as intake minus total faecal excretion of the isotopes over 68-92 h after administration. The fractional apparent absorption values for Zn and Ca were 33.9 (SD 164) % (range 19.2-639 %) and 53.5 (SD 12.6) %) (range 36.7-71.7 %) respectively. Re-excretion of absorbed 70Zn (> 68-92 h to 21 d after intake of the labelled meal) was 044 (SD 038) %] of administered dose while only one infant re-excreted detectable amounts of 42Ca (1.74%) of administered dose). The analysis of individual stool samples confirmed that 72 h is a sufficient time period for complete collections of non-absorbed isotopes in faecal material from infants during the weaning period and that re-excretion of initially absorbed 70Zn and 42Ca (> 68-92 h to 21 d after intake of the labelled meal) is negligible.

To study the effect on plasma 25-hydroxycholecalciferol (25(OH)D), 1,25-dihydroxycholecalciferol (1,25(OH)2D) and parathyroid hormone (PTH) we supplemented premenopausal (aged 30 (SD 7) years) and postmenopausal (aged 61 (SD 2) years) white women living in The Netherlands in late winter/early spring, and elderly black and white women (aged 75 (SD 6) years) living in Curacao (Dutch Antilles) with either 10 or 20µpg cholecalciferol/d for 4, 5 and 9 weeks respectively. Baseline plasma 25(OH)D concentration of Dutch women was lower than that of Curacao women. Postmenopausal Dutch women had a higher PTH concentration in plasma than premenopausal Dutch and postmenopausal Curaçao women. There were no differences in plasma 1,25(OH)2D. Cholecalciferol administration increased 25(OH)D in all groups, 1,25(OH)2D in postmenopausal Curaçao women and PTH in postmenopausal Curaçao women and premenopausal Dutch women. Serum and urinary Ca and phosphate concentrations did not change. There were no response differences between 10 and 20 µg doses. Oral cholecalciferol administration (either 10 or 20 µg/d) to women living at northern latitudes in late winter/early spring increased 25(OH)D levels to the baseline levels of elderly people living in the tropics.

The effect of fermentation on colonic absorption of Ca and Mg was investigated in 8-week-old rats adapted to diets containing either digestible wheat starch (DS diets) or including resistant starch, i.e. 350 g raw potato starch/kg (RS diets). The dietary Ca level of the DS and RS diets was 2·5 or 7·5 g/kg. RS diets resulted in enlargements of the caecum together with hypertrophy of the caecal wall. Acidification of the caecal contents by mictobial fermentation of RS was influenced by the dietary Ca level. Very acidic pH conditions and relatively low concentrations of short-chain fatty acids, in the presence of lactic acid fermentation, were observed with the 2·5 g Ca/kg level. Rats fed on RS diets had a bhigher pencentage of soluble Ca (and inorganic phosphate) in the caecum, particularly of rats adapted to the high Ca level. As a result of the hypertrophy of the caecal wall and of an elevated concentration of soluble Cas, the caecal absorption of Ca was 5-6 fold higher in the RS groups than in the DS groups. The difference between dietary intakle and faecal excretion (DI-FE) of Ca was higher in rats fed on RS diets than in those fed on DS diets, when the dietary Ca level was 2·5 g/kg. With the higher Ca intake the elevated rate of Ca absorption from the caecum in RS-fed rats was not paralleled by an enhanced DI-EE difference: this suggests a shift of the Ca absorption towards the large intestine. Feeding Rs diets also enhanced Mg caecal absorption, resulting in the substatntially higher DI-FE difference for Mg, especially with the 2·5 g Ca/kg diets, because a high Ca intake tends to inhibit Mg absorption. The present findings support the view that the large intestine may represent a major site of Ca (and Mg) absorption when acidic fermentations take place. This process could improve the digestive Ca balance when the dietary Ca supply is low; when the Ca supply is affluent, it rather shits Ca absorption towards a more distal site fo the digestive tract.

The purpose of the present study was to determine the effects of feeding nutritionally adequate and increased levels of vitamin A (retinyl acetate at 1·4, 34·4, and 206·4 mg/kg diet) in combination with adequate or increased Zn (12 and 240 mg/kg) and Cu (5 and 50mg/kg) on serum and tissue concentrations of retinol and retinyl palmitate and on indices of Cu and Zn status in female Sprague–Dawley rats, and to measure interactive effects of such nutrient imbalances. Rats fed on diets containing 34·4 and 206·4 mg vitamin A/kg had higher feed intakes and relative Liver weights than those fed on diets containing 1.4mg vitamin A/kg. An interaction between dietary Cu and Zn and an independent effect of vitamin A affected serum ceruloplasmin oxidase (EC 1.16.3.1) activity. Rats fed on high Zn, adequate-Cu diets (240 and 5 mg Zn and Cu/kg respectively) had lower serum ceruloplasmin oxidase levels than rats fed on adequate-Zn, adequate-Cu diets (12 and 5 mg Zn and Cu/kg respectively). This effect was not observed in rats fed on high-Zn, high-Cu diets (240 and 50mg Zn and Cu/kg respectively). Alterations in dietary levels of Cu and vitamin A independently affected haemoglobin levels. Serum cholesterol concentration was affected by interactions between Zn and vitamin A and Cu and vitamin A. Levels of retinol and retinyl palmitate in liver and kidney were significantly higher in rats fed on diets with increased dietary vitamin A than in those fed on diets with adequate vitamin A. Three-way interactions among Cu, Zn, and vitamin A affected levels of retinol in serum and liver. Two-way interactions between Cu and vitamin A affected liver retinyl palmitate and the sum of liver retinol + retinyl palmitate. An independent effect of dietary Zn on these variables was also observed. Interactions between Cu and vitamin A affected levels of Cu in liver and kidney, while Fe and Zn in kidney were affected by interactions between Cu and Zn. This study demonstrates that differing interactions among variables of vitamin A metabolism and mineral status occur with higher dietary levels of vitamin A, Zn and Cu in the rat

We developed a new type of anal cup for prevention of coprophagy and determined whether the absorption of Ca and Mg and the stimulatory effects of feeding fructo-oligosaccharides (FO) on theabsorption of Ca and Mg were altered by prevention of coprophagy in rats. Rats were fed on a FO-free diet or a diet containing 50 g FO/kg for 2 weeks with or without prevention of coprophagy. FO-feeding increased the apparent absorptive ratio of Ca and Mg in rats with or without prevention of coprophagy. However, in the FO-fed groups the absorptive ratio of Mg in rats with prevention of coprophagy was higher than in rats withont prevention of coprophagy. The Ca content of the femur was higher in rats fed on the FO-diet than in rats fed on the FO-free diet both with and without coprophagy. In conclusion, FO-feeding increased the absorption of Ca and Mg in rats both with and without coprophagy. Moreover, prevention of coprophagy enhanced the absorption of Mg in rats fed with FO. Coprophagy has to be considered when the effects of luminal fermentation or mineral absorption are examined in rats

The purpose of the present study was to assess the role of the liver in the plasma-cholesterol-lowering effect of soyabean lecithin. Normolipidaemic rats were fed on lecithin-enriched or control diets with the same amount of protein. The lecithin diets contained 200 g/kg high-fat commercial semi-purified soyabean lecithin (230 g/kg total lipids as soyabean phosphatidylcholine) or 200 g/kg high-fat purified soyabean lecithin (930 g/kg total lipids as soyabean phosphatidylcholine). The control diets were a low-fat diet (40 g fat/kg) and a high-fat triacylglycerol-rich diet (200 g fat/kg). The high-fat diets were isoenergetic. The cholesterol-lowering effect of the lecithin-enriched diets was associated with significantly lower levels of plasma total- and HDL-cholesterol and significantly higher levels of bile phosphatidylcholine (PC), bile salts and cholesterol. These findings suggest that the liver plays a major role in the reduction of plasma cholesterol, the increased biliary lipid being provided by both HDL and the hepatic microsomal pools of PC and cholesterol.

Cu has long been known to influence immune responses. An in vitro model system was established in which human myeloid(HL-60), B-lymphoid (Raji) and T-lymphoid (Molt-3) cell lines could be grown in culture media of varying Cu levels. Initially Cu was removed from the medium by dialysisof fetal calf serum against a metal-ion chelator, minor depletion of other trace metals being obviated by repletion with appropriate metal salts. The growth rate of HL-60 was significantly (P<0·05) inhibited by 72 h Cu depletion. Molt-3 cells required a longer period, up to 144 h, in Cudepleted medium before growth was impaired. Raji-cell growth was not affected. These results confirmed clinical observations that T-cell functions were more sensitive to Cu deprivation than B cells. Analysis of intracellular metal levels in Molt-3 cells showed that Cu levels had been significantly lowered (P <0·05) although Ca2+ levels were raised. Intracellular activity of the antioxidant enzyme superoxide dismutase (EC 1. 15. 1. 1) was significantly impaired (P<0·05) in Molt-3 cells grown in Cudepleted medium. Activity of the mitochondria1 enzyme cytochrome c oxidase (EC 1. 9. 3. 1) was also significantly impaired (P <0·05) by Cu depletion. Each of these findings indicates an increase in the potential for cellular damage by reduced antioxidant activity, impairment of normal mitochondrial activity and excessive Ca2+influx. A major consequence of the type of damage occurring under these circumstances is membrane disruption. This was confirmed by scanning electron microscopy of Molt-3 cells grown under varying Cu levels.

The influence of high CaCO3, intake on the bioavailability of Fe from FeSO4, was assessed during Fe repletion of rats with Fe-deficiency-induced anaemia. Fe-deficient rats with a mean blood haemoglobin concentration of 4. 1 mmol/l were fed on purified Fe-adequate diets containing either 6.2 or 25.0g CaCO,/kg (ten rats per group). Haemoglobin repletion after 14 d was significantly depressed by high CaCO, intake (9.5 v. 9.8 mmol/l for high and low CaCO, intake respectively; P = 0·03), as was apparent Fe retention (367 v. 552 µg/d during days 5–7, P<0·001; 146 v. 196 µg/d during days 19–21, P < 0·001). The concentration of Fe in the liquid phase of the proximal half of the small intestine was significantly lower in the high-CaCO, group (3.71 v. 520 µg/g digesta; P = 0-02). Mucosal uptake and mucosal transfer of Fe were determined with orally administered 59Fe and Cr as a non-absorbable marker. Mucosal transfer was significantly diminished by CaCO, loading (90 v. 100% of mucosal uptake; P = 0·04), whereas mucosal uptake was not. 59Fe retention values at 14 d after administration were not significantly different (57.6 v. 51.9%; P = 0·14). Fe contents of liver and spleen were significantly decreased by high compared with low CaCO, intake (879 v. 590 pg Fe in liver, P < 0·001; 92 v. 63pg Fe in spleen, P <0·001). It is concluded that high intake of CaCO, depresses Fe bioavailability in rats. The CaCO,-induced decrease in Fe solubility in the digesta probably was associated with an increased efficiency of mucosal Fe uptake so that the amount of mucosal uptake remained unaltered. The CaCO,-induced decrease in Fe transfer through the mucosal cytoplasm and/or basolateral membrane may have been responsible for the concurrent decrease in Fe bioavailability.

To clarify the relationship between intracellular concentrations of methylmalonic acid and metabolic and growth inhibition in vitamin B12-deficient rats, hepatic methylmalonic acidlevels were assayed and inhibition of glucose and glutamic acid metabolism by methylmalonic acid was studied in isolated hepatocytes. Vitamin B12-deficient rats (14 weeks old) excreted more urinary methylmalonic acid and had lower body weights than the control rats. Hepatic methylmalonic acid levels (3·6 (SD 1·30)–5·3 (SD 0·51) µmol/g tissue; 7·9 (SD 2·90)–11·8 (SD 1·14) mM) were increased and correlated with the extent of the growth retardation during vitamin B12-deficiency. Isolated hepatocytes and mitochondria from normally fed rats were labelled with [14C(U)]glucose and [14C(U)]glutamic acid respectively, in the presence or absence of 5mM-methylmalonic acid. Although methylmalonic acid did not affect the incorporation of 14C into protein and organic acid fractions in the hepatocytes, it inhibited 14CO2 formation (an index of glucose oxidation by the Krebs cycle) by 25% and incorporation of 14C into the amino acid fractionby 30%. In the mitochondria, methylmalonic acid inhibited 14CO2, formation (indicating glutamic acid oxidation by the Krebs cycle) by 70%, but not the incorporation of 14C into the protein fraction. The incorporation of 14C into the organic acid fraction was significantly stimulated by the addition of methylmalonic acid. These results indicate that the unusual accumulation of methylmalonic acid caused by vitamin B12-deficiency disrupts normal glucose and glutamic acid metabolism in rat liver, probably by inhibiting the Krebs cycle.

In order to establish a method usable in the field for diagnosis of pre-clinical vitamin A deficiency, conjunctival impression cytology with transfer (CICT) was used in 200normal slum children aged 6−120 months in Chandigarh, India. Conjunctival impressions taken on cellulose acetate paper were transferred to glass slides which were fixed in ethanol and stained with alcian blue and carbol fuchsin. Sixty samples of cellulose acetate paper were preserved after transfer of impression and were stained later. Transfer was complete in forty-eight samples and in the other twelve it was incomplete. Conjunctival impressions on paper and slide of these twelve cases were comparable. At a plasma retinol concentration of <0·70 pmol/l measured by HPLC the sensitivity, specificity and positive predictive values of CICT were 90·5%, 100% and 100% respectively. Compared with conventional conjunctival impression cytology, CICT is less time consuming, cheaper and comparable in validity. It is thus more suitable than the conventional method for mass screening

The effects of dietary fat and dietary fibres on blood pressure, serum lipids and platelet aggregation in spontaneously hypertensive and Wistar-Kyoto rats have been investigated. The systolic and diastolic blood pressure values were increased with increasing amounts of dietary fat and normalized by dietary fibre supplementation. The greatest reduction in blood pressure was obtained by a combination of reduced dietary fat and supplementary dietary fibre. Addition of dietary fibre when the amount of dietary fat was high or reduction of dietary fat when the amount of dietary fibre was low gave a smaller effect. In both rat strains the decreases in systolic and diastolic blood pressure values after reducing dietary fat and/or increasing dietary fibre were about 10-15 mmHg. Serum total cholesterol and serum HDL-cholesterol concentrations were reduced by reduction of dietary fat or a combination of dietary fat reduction and dietary fibre addition. A combination of dietary fat reduction and dietary fibre addition was the most effective dietary change for reducing serum triacylglycerol concentration and platelet aggregation. The present study demonstrates that in this experimental model it is possible to reduce risk indicators of coronary heart disease significantly by changing dietary habits.

We measured the vitamin K status in postmortem human tissues (brain, heart, kidney, liver, lung, pancreas) to see if there is a tissue-specific distribution pattern. Phylloquinone (K1,) was recovered in all tissues with relatively high levels in liver, heart and pancreas (medians, 10·6 (4·8), 9·3 (4·2), 28·4 (12·8) pmol(ng)/g wet weight tissue); low levels (< 2 pmol/g) were found in brain, kidney and lung. Menaquinone-4 (MK-4) was recovered from most of the tissues; its levels exceeded the K1 levels in brain and kidney (median, 2·8 ng/g) and equalled K1 in pancreas. Liver, heart and lung were low in MK–4. The higher menaquinones, MK-6–11, were recovered in the liver samples (n 6), traces of MK-6–9 were found in some of the heart and pancreas samples. The results show that in man there are tissue-specific, vitamin-K distribution patterns comparable to those in the rat. Furthermore, the accumulation of vitamin K in heart, brain and pancreas suggests a hitherto unrecognized physiological function of this vitamin.