To investigate the mechanism of tissue Fe accumulation in graded Mg deficiency rats were fed on diets of different Mg contents (70, 110, 208, 330, and 850 mg Mg/kg) for 10, 20, and 30 d during rapid growth. There was no significant impact of Mg deficiency or high luminal Mg concentrations on intestinal 59Fe transfer in vitro or in vivo. Plasma Mg concentrations and body weight started to decrease after 10 d. Significant haemolytic anaemia was observed after 20 d with siderosis in liver and spleen developing in parallel. Anaemia showed no features of Fe deficiency or infiammation. Comparison between the 70 mg Mg/kg group and animals that received the same quantity of a Mg-adequate diet (850 mg Mg/kg) permitted estimation of quantities of Fe liberated by haemolysis and the increased Fe content in liver and spleen. Both variables showed a high degree of correlation, indicating that the excess of liberated haemoglobin Fe was stored in the tissue. The erythropoietic activity was high during rapid growth, i.e. at days 10 and 20 and decreased significantly after 30 d in all except the most Mg-deficient groups. However, haemolytic anaemia developed because even the high erythropoietic activity in the 70 and 110 mg Mg/kg groups was not sutlicient to recycle all haemoglobin Fe liberated by haemolysis. After 30 d of Mg-deficient feeding the erythrocyte Mg content had decreased to 40% of control values. According to the literature Mg-deficient erythrocytes have a decreased survival time which is likely to be the cause of the observed haemolysis.

Diets containing unheated casein (CD; control) or a casein-glucose mixture (CGD) previously heated at 140° for 2 h were fed to two groups of young rats for 21 d. Differences in body weight, feed consumption, thymus and spleen growth, protein metabolism and in vivo immune response were then determined. For this last experiment, animals were inoculated with sheep erythrocytes (SRBC) on day 15 to provide an immunological challenge. No changes were seen in body weight, feed consumption or feed conversion ratios. Neither were significant differences found in spleen weight, protein content, DNA content, DNase (EC 3.1.4.6) activity or lymphocyte count, suggesting that spleen cell growth remained similar in all the animals studied. The CGD induced marked increases in thymus DNA content whilst the protein:DNA ratio became lower. Spleen RNA content was similar in all rats, but thymus RNA content was 29% lower in the CGD group, although this difference did not reach statistical significance. This fact might be a consequence of the low RNase (EC 2.7.7.16) activity and RNase:RNA ratios in the thymus glands of CGD-fed animals. Further, the number of splenic plasma cells secreting anti-SRBC antibodies (direct plaque-forming cells) was significantly decreased in the same group. It might be concluded that both diets are adequate for rat growth and that the differences observed in the thymus of CGD-fed rats may be directed towards preserving tissue function. Nevertheless, the CGD did cause immunological disturbances affecting the humoral immune response.

To explore whether the metabolic responses to trans, compared with cis, fatty acids depend on the source of dietary polyunsaturated fatty acids (PUFA), male Sprague-Dawley rats, 5 weeks old, were fed on diets containing 30 g oleic (cis) or elaidic (trans) acids/kg in combination with either 70 g perilla oil (α-linolenic acid) or safflowerseed oil (linoleic acid)/kg for 3 weeks in separate experiments. The dietary fats were adjusted to have the same level of total PUFA. The dietary manipulation did not influence the growth indices, but spleen weight was greater when the dietary PUFA source was perilla oil. The incorporation of trans fatty acid into liver phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylserine and adipose tissue lipids, particularly phospholipids, was significantly higher when rats were fed on safflowerseed oil compared with perilla oil. However, only limited differences were observed in the effects of cis and trans fatty acids on the proportions of PUFA in liver phospholipids. Splenic production of prostaglandin E2 was reduced by trans fatty acid when safflowerseed oil was the PUFA source, but no trans effect was observed on leukotriene C4 production. Dietary PUFA significantly influenced the concentration of plasma immunoglobulins (Ig) but the effect of geometry was only seen in IgG which was increased by trans acid. Dietary trans fatty acid increased the CD4+: CD8+ T-lymphocyte ratio in the spleen, reflecting a decreasing trend of the proportion of CD8+, when combined with perilla oil. These observations indicate that the type of PUFA simultaneously ingested specifically infiuences the effect that trans acid exerts on PUFA metabolism, eicosanoid production and some immune indices.

The present work aimed to investigate the feasibility of using stable isotopes and inductively-coupled plasma mass spectrometry (ICP/MS) to study Mg absorption in rats. Male Wistar rats, aged 7 weeks and weighing 180g, were used. They were fed on a semi-purified diet containing 1070mg Mg/kg, and had free access to feed and distilled water. In the first experiment, after a 16d adaptation period, two doses of enriched 25Mg (6 and 12mg) were administered by oral intubation, faeces and urine were collected daily and blood was sampled. Isotope ratios were determined by ICP/;MS. ‘True’ absorption values, using the faecal isotope data, were 0.63 and 0.56 in rats receiving 6 and 12mg 25Mg respectively, while apparent absorption was 0.50 for two successive periods of metabolic balance studies. Moreover, the oral isotope administration resulted in a measurable isotopic enrichment in plasma within hours which was still detectable on the third day following the isotope administration. In the second experiment, investigating the double labelling technique, similar rats were dosed Simultaneously with 5mg 26Mg orally (premixed with diet) and 0.29mg 25Mg intravenously. The calculated Mg true absorption values were very similar when calculated from blood or urine data (0.38) but were lower than that obtained from faecal data (0.50). The possible causes of such an unexpected difference and limits of the double labelling technique for Mg absorption are discussed here. Together these results indicate that although 25Mg and 26Mg isotopes have high natural abundance, the described methodology permits meaningful investigations of Mg bioavailability and metabolism

Studies in autoimmune-prone NZB/W F1 mice have demonstrated that the amount of dietary fat can affect autoantibody production and the disease course of autoimmune diseases. Anti-cardiolipin antibodies have been found to play a major role in thrombus formation and the increase of abortion rate in both human lupus patients and murine lupus. The present study investigated further the effect of dietary fat on lipid and anti-cardiolipin antibody production in autoimmune-prone mice. Two groups of NZB/W F1 mice were fed on diets containing 200 g dietary fat/kg and 50 g dietary fat/kg respectively, the fat being composed of equal amounts of lard and soyabean oil. Serum levels of lipids, immunoglobulin (Ig) anti-single stranded DNA and anti-cardiolipin antibodies were followed regularly every month and mice were killed for in vitro experiments after 5 months on the experimental diets. The results showed that serum triacylglycerol concentration was lower in mice fed on the high-fat diet than in those fed on 50 g fat/kg. There was no significant difference in hepatic lipid contents; however, the fatty acid contents were different between these two groups. Hepatic linoleic acid (18:2n−6) and arachidonic acid (20:4n−6) concentrations were higher in mice fed on the high-fat diet. There were no significant differences in serum IgM concentrations or IgM anti-cardiolipin antibody levels between these two groups. However, IgG anti-cardiolipin antibody levels were higher in mice fed on the high-fat diet at the age of 3–4 months. Total serum IgG concentration was noted to be higher, but in contrast, serum IgA was lower, in the high-dietary-fat group. These findings suggest that high dietary fat may affect lipid metabolism and autoantibody levels in autoimmune diseases

The purpose of this study was, first to explore metabolic interactions between Cu and ascorbic acid in guinea-pigs, particularly with respect to any possible disadvantages of high ascorbatein the presence of low Cu intakes, and second, to test the hypothesis that variations in ascorbate and/or Cu status might inhence collagen cross-linking, either by inducing a change in thecross-links: hydroxyproline ratio, or by inducing a change in the pyridinoline: deoxypyridinoline cross-linls ratio. Four matched groups, each of eight male weanliig Dunkin-Hartley guinea-pigs, were maintained on purified diets containing either no added Cu, or 150 mg Cu/kg diet, and either 0·1 g or 30 g ascorbic acid/kg diet. They were then killed 8 weeks later, and the following indices were measured body and organ weights; blood haemoglobin; adrenal ascorbate concentrations; Cu concentrations in plasma, liver and femur; superoxide dismutase (EC 1.15.1.1) activity in whole blood and liver; hydroxyproline, pyridinoline and deoxypyridinoline in femur and in urine. The principal observations were: Cu intake significantly affected blood and tissue Cu concentrations and superoxide dismutase activity; and ascorbic acid intake significantlyaffected adrenal ascorbate levels and the deoxypyridinoline: pyridinoline cross-links ratio, especially in bone (femur). There was evidence of a significant interaction between ascorbateand Cu with respect to adrenal and plasma Cu concentrations, blood superoxide dismutase activityand body weights. We conclude that interactions between ascorbate and Cu at the functional level were present but modest, and that a new and potentially powerful functional index of ascorbate status may exist within the deoxypyridinoline: pyridinoliie collagen cross-link ratio.

The efficiency with which β-carotene protects against oxidative stress in chicken embryo fibrobiasts (CEF) at low O2 partial pressures was assessed. Primary cultures of CEF were grown at low O2 partial pressures and oxidatively stressed by exposure to paraquat (PQ). Activities of the antioxidant enzymes superoxide dismutase (EC 1.15.1.1; SOD), catalase (EC 1.11.1.6; CAT) and glutathione peroxidase (EC 1.11.1.9; GSH-Px) were measured as indices of oxidative stress. CEF incubated with 0·25–1·0 mM—PQ for 18 h exhibited increased SOD and CAT activities compared with non-PQ-treated control cells (P < 0·001). No cytotoxicity as indicated by lactate dehydrogenase (EC 1.1.1.27; LDH) release was observed at PQ concentrations below 2·0 mM. Incorporation of added β-carotene into 0·25 mM-PQ-treated cells prevented the PQ-induced increases in SOD and CAT, and activities were similar to those seen in non-PQ-treated control cells. GSH-Px activity decreased relative to its control value on exposure to 0·25 mM-PQ and β-carotene prevented this decrease in a dose-dependent manner. The proportion of LDH released from the CEF treated with β-carotene remained below the control value of 2·5% at all times.

In work aimed at developing methodologies for validation of estimates of the dietary intake of free-living individuals, Bingham and colleagues have examined the use of urinary nitrogen (UN) excretion as an index of protein intake (Bingham & Cummings, 1985; Bingham, 1994; Bingham et al. 1995). The basis of this approach is that in subjects in N equilibrium, N intake is assumed equal to N excretion. Thus, if N excretion is measured it should indicate N and dietary protein intakes. In practice, since most N excretion occurs via the urine, which is relatively simple to collect, Bingham and colleagues examined the relationship between 24 h UN and N intake (DN). They argued that in subjects on typical UK diets UN should bear a fixed relationship to DN, and measured it carefully in a group of subjects. They showed that in a group of eight individuals in which intake and N excretion were measured meticulously, UN was 81% DN (SD 2, range 78–83%). They argued that measurement of this ratio could be used to assess the validity of food intake measurement and concluded ‘In healthy individuals eating normal western diets, 24 h urine N from an 8 day collection verified for its completeness by the PABA check method, should establish urine N to within 81(SD 5)% of the habitual dietary intake, range 70–90%. If the dietary assessment from 18 days of records or 24 h recalls or the diet history falls within these limits, it can be stated that there is no evidence of interference with normal dietary habits, or of reporting errors….’ (Bingham & Cummings, 1985).

Vitamin E is the major lipid-soluble antioxidant in human subjects and is crucial in protecting polyunsaturated fatty acids (PUFA) against lipid peroxidation. Dietary PUFA have been suggested to inhibit the absorption of vitamin E. The present study in young male rats was designed to investigate the effect of increasing concentrations of dietary linoleic acid on the faecal excretion of vitamin E. The rats were fed on semi-synthetic diets containing two concentrations of fat (59 g/kg diet, 15 energy % (en%) or 131 g/kg, 30 en%) for 3 weeks. Triacylglycerol rich in linoleic acid was added at the expense of triacylglycerol rich in saturated fatty acids to obtain dietary concentrations of 13,39 or 66 g linoleic acid/kg diet for the high-fat diet (131 g fat/kg) and 12, 24 or 36 g linoleic acid/kg diet for the reduced-fat diet (59 g fat/kg). The results from the present study demonstrate that the faecal excretion of vitamin E was significantly lower in rats fed on diets with high levels of linoleic acid compared with rats fed on lower levels of linoleic acid irrespective of the dietary fat content. The concentration of vitamin E in liver and plasma was significantly lower in animals fed on the highest concentration of linoleic acid compared with those fed on the lowest level. Results from the present study also demonstrate that at the same concentration of linoleic acid, the faecal excretion of vitamin E in rats fed on reduced-fat diets was significantly lower than in rats fed on high-fat diets. Our findings indicate that the apparent absorption of vitamin E is not inhibited by dietary PUFA. Results from the present study also demonstrate that a reduction of dietary fat content from 30 en% to 15 en% does not lower the apparent absorption of vitamin E