A thyroid-hormonal evaluation of thirty-five women consuming commercially packed milk containing thiocyanate was carried out. The mean serum thiocyanate concentration, which was measured by the FeCl3 colour test, was significantly higher (P < 0.01) than that of control subjects. Serum thyroxine (T4), triiodothyronine (T3) and thyroid-stimulating hormone (TSH) concentrations of exposed women were compared with those of thirty-five control subjects. Thiocyanate ingestion was associated with lower levels of T4 (P <0.01) and higher levels of TSH (P < 0.01) compared with the control subjects. T3 was found to be higher in the women consuming thiocyanate-containing milk but the difference was not significant. The serum T4 level was found to be negatively correlated (r −0.359, P <0.05) while the TSH level was positively correlated (r 0.354, P < 0.05) with thiocyanate concentration in the exposed group. From this study, it appears that ingestion of milk with added thiocyanate impairs thyroid function.

The effect of long-term differences in diet composition on whole-body acid–base status was examined in thirty-three young healthy females. The volunteers were recruited from two separate groups matched approximately for age, height and weight; one group regularly ate meat (omnivores; n 20) and one group did not (vegetarians; n 13). All subjects completed a 7 d weighed intake of food, and from their dietary records, total energy, carbohydrate (CHO), fat and protein content were estimated using computer-based food composition tables. During this week they reported to the laboratory on two occasions, following an overnight fast and separated by at least 48h. Arterialized venous blood samples were obtained on each visit and these were analysed for blood acid–base status. Haemoglobin and packed cell volume, serum total cholesterol and HDL-cholesterol, serum albumin and total protein were also determined. Two 24 h urine collections were completed; the volume was recorded and samples were analysed for pH, titratable acid and Mg and Ca concentration. Total energy intake of the omnivores was greater (P = 0.0003) than that of the vegetarian group. Dietary intake of CHO (P = 0.024), fat (P = 0.0054) and protein (P = 0.0002) were higher in the omnivorous group than in the vegetarians. There were no differences between the two groups with respect to blood CO2 partial pressure, plasma HCO3- and blood base excess, but blood pH was slightly higher in the omnivores (P = 0.064). Measures of urine acid–base status suggested a lower pH in the omnivore group, but this difference was not statistically significant; a greater titratable acid output was observed with the omnivorous group compared with the vegetarians (48.9 (se 20.3) ν. 35.3 (se 23.3) mEq/24h; P = 0.018). Although the dietary intake of Ca was not different between the two groups, urinary Ca excretion of the omnivores was significantly higher (3.87 (sd 1.34) ν. 3.22 (sd 1.20) mmol/24h) than that of the vegetarians (P = 0.014). It is suggested that the higher protein intake of the omnivores resulted in an increase in urinary total acid excretion, which may explain the higher rate of Ca excretion.

Free-living energy expenditure was estimated by doubly-labelled water (DLW) and continuous heart-rate (HR) monitoring over nine consecutive days in nine healthy men with sedentary occupations but different levels of leisure-time physical activity. Individual calibrations of the HR-energy expenditure (EE) relationship were obtained for each subject using 30 min average values of HR and EE obtained during 24h whole-body calorimetry with a defined exercise protocol, and additional data points for individual leisure activities measured with an Oxylog portable O2 consumption meter. The HR data were processed to remove spurious values and insert missing data before the calculation of EE from second-order polynomial equations relating EE to HR. After data processing, the HR-derived EE for this group of subjects was on average 0.8 (sem 0.6) MJ/d, or 6.0 (sem 4.2)% higher than that estimated by DLW. The diary-respirometer method, used over the same 9d, gave values which were 1.9 (sem 0.7) MJ/d, or -12.1 (sem 4.0)% lower than the DLW method. The results suggest that HR monitoring can provide a better estimate of 24 h EE of groups than the diary-respirometer method, but show that both methods can introduce errors of 20% or more in individuals.

The aim of the present study was to validate against the doubly-labelled water (DLW) technique the factorial method and the heart rate (HR) recording method for determining daily energy expenditure (DEE) of elderly people in free-living conditions. The two methods were first calibrated and validated in twelve healthy subjects (six males and six females; 70·1 (sd 2·7) years) from opencircuit whole-body indirect calorimetry measurements during three consecutive days and during 1 d respectively. Mean energy costs of the various usual activities were determined for each subject using the factorial method, and individual relationships were set up between HR and energy expenditure for the HR recording method. In free-living conditions, DEE was determined over the same period of time by the DLW, the factorial and the HR recording methods during 17, 14 and 4 d respectively. Mean free-living DEE values for men estimated using the DLW, the factorial and the HR recording methods were 12·8 (sd 3·1), 12·7 (sd 2·2) and 13·5 (sd 2·7) MJ/d respectively. Mean free-living DEE values for women were 9·6 (sd 0·8), 8·8 (sd 1·2) and 10·2 (sd 1·5) MJ/d respectively. No significant differences were found between the three methods for either sex, using the Bland & Altman (1986) test. Mean differences in DEE of men were -0·9 (sd 11·8) % between the factorial and DLW methods, and +4·7 (sd 16·1) % between the HR recording and DLW methods. Similarly, in women, mean differences were -7·7 (sd 12·7) % between the factorial and DLW methods, and +5·9 (sd 8·8) % between the HR recording and DLW methods. It was concluded that the factorial and the HR recording methods are satisfactory alternatives to the DLW method when considering the mean DEE of a group of subjects. Furthermore, mean energy costs of activities calculated in the present study using the factorial method were shown to be suitable for determining free-living DEE of elderly people when the reference value (i.e. sleeping metabolic rate) is accurately measured.

Even-numbered dicarboxylic acids (DA) have been proposed as an alternative fuel substrate in parenteral nutrition. In particular, dodecanedioic acid (C12) shows a rapid plasma clearance from tissues, a very low urinary excretion compared with other DA and a high oxidation rate. The aim of the present study was to investigate the effect of C12 infusion on insulin-stimulated glucose uptake in patients with non-insulin-dependent diabetes mellitus (NIDDM) compared with healthy volunteers. A primed-constant infusion of C12 (0·39 mmol/min) was administered over 240 min, and at 120 min a 2 h euglycaemic hyperinsulinaemic clamp was performed. Blood specimens were sampled every 30 min and fractioned urines were collected over 24 h. The levels of C12 were measured by HPLC. Indirect calorimetry was performed continuously during the entire session. Body composition was assessed in all subjects studied to obtain fat-free mass (FFM) values. Whole-body glucose uptake decreased significantly during C12 infusion in both groups, although this effect was much more evident (P < 0·01) in NIDDM patients (52·4 (sd 15·8) % decrease compared with saline) than in controls (25·9 (sd 12·1) % decrease). The M value (μmol/kgffm per min) was reduced by C12 to lower levels in NIDDM patients than in normal controls (12·6 (sd 3·9) ν. 25·9 (sd 4·5), P < 0·01). Urinary excretion of C12 over 24 h was significantly lower in NIDDM patients than in controls (4·26 (sd 0·30) mmol ν. 5·43 (sd 0·48), P < 0·01), corresponding to less than 3 % of the administered dose. The infusion of C12 decreased non-protein RQ significantly in both groups of patients. In conclusion, this study shows, for the first time, that C12 significantly reduces glucose uptake in both normal controls and NIDDM patients, although this sparing effect on glucose uptake is much more pronounced in diabetic patients. These data suggest that C12 decreases glucose uptake and oxidation, mainly through a mechanism of substrate competition. Thus, it might be a useful alternative substrate in enteral or parenteral nutrition, sparing glucose utilization and increasing glycogen stores, in those clinical conditions, like NIDDM, where reduced insulin-induced glucose uptake and oxidation are observed.

In a double-blind, placebo-controlled study, thirty-three subjects were allocated to undergo either a 4-week treatment with oral Mg supplementation (Mg(OH)2; 411–548 mg Mg/d) or a placebo. The urinary excretion of Mg increased significantly in both the first 2 weeks and the following 2 weeks of Mg supplementation, while the urinary Na excretion also increased significantly over the experimental period. The systolic and diastolic blood pressure values decreased significantly in the Mg group, but not in the placebo group. The urinary aldosterone excretion and packed cell volume increased significantly during the last 2 weeks of the experimental period compared with the run-in period and first 2 weeks of supplementation. There was a statistically significant positive correlation between the values for urinary noradrenaline excretion and diastolic blood pressure at the end of the supplementation period (both expressed as a percentage of the run-in value). Statistically significant increases in lecithin-cholesterol acyltransferase (EC 2.3.1.43; LCAT), HDL-cholesterol and apolipoprotein AI were also observed after Mg supplementation. A significant positive correlation was observed between the levels of LCAT and urinary Mg excretion for the experimental period (expressed as a percentage of the run-in value). The total cholesterol: HDL-cholesterol ratio decreased significantly during the last 2 weeks of Mg supplementation compared with the first 2 weeks and the run-in periods, but this did not occur in the placebo group. These results suggest that Mg supplementation may lower blood pressure through the suppression of the adrenergic activity and possible natriuresis, while also improving the serum lipids through the activation of LCAT in human subjects.

To investigate Fe nutritional indices in malaria infection in children, haematology (blood haemoglobin, plasma ferritin, transferrin, Fe, and transferrin saturation), acute phase markers (albumin and caeruloplasmin) and liver function tests were studied in fifty consecutive cases of severe and mild falciparum malaria, fifty matched controls and twenty-three cases of asymptomatic malaria. Blood haemoglobin and transferrin were lower, while ferritin and transferrin saturation were higher, in groups with symptomatic malaria in comparison with the control group. The differences were greatest with the severest form of the disease. There were no differences between any of the groups in plasma Fe. Plasma transferrin correlated directly with albumin in asymptomatic, mild and severe malaria groups (r 0.48, 0.65 and 0.83; P < 0.05, P < 0.01 and P < 0.001 respectively), and inversely with caeruloplasmin (r −0.65, −0.34 and −0.43; P < 0.01, P < 0.05 and P < 0.01 respectively). For ferritin, the correlation was inverse with albumin (r −0.65, −0.57 and −0.64; P < 0.01, P < 0.001 and P < 0.001 respectively and direct with caeruloplasmin (r 0.83, 0.21 and 0.49, P < 0.001, NS and P < 0.001 respectively). Multiple regression analysis on data from all patients combined indicated that albumin, and to a lesser extent alanine aminotransferase (EC 2.6.1.2) activity, explained 62 % of the variance in transferrin. Caeruloplasmin, parasite count and albumin explained 59 % of the variance in ferritin, and transferrin and unconjugated bilirubin explained 62 % of the variance in Fe values. In conclusion, these data suggest that low transferrin and high ferritin values are primarily due to the acute phase response. High transferrin saturation and lack of differences in plasma Fe between the groups are probably due to Fe released from lysed erythrocytes. Finally, in both symptomatic and asymptomatic malaria, indices of Fe status can be misleading and may be especially problematic in community studies in malaria-endemic areas where asymptomatic malaria may be common.

It has been suggested that decreased immune responsiveness in the elderly may be counteracted by the antioxidant vitamin E. In a 3-month double-blind placebo-controlled intervention trial among elderly subjects aged 65 years and over we studied the effects of a daily dose of 100 mg dl-α-tocopheryl acetate on the cellular immune responsiveness (n 52) measured by the in vitro response of peripheral blood mononuclear cells (PBMC) to the mitogens concanavalin A (ConA) and phytohaemagglutinin (PHA). Also effects on the humoral immune responsiveness (n 74) were investigated by measuring immunoglobulin (Ig)G, IgG4 and IgA antibody concentrations against various common antigens. In the vitamin E group plasma α-tocopherol increased by 51 % (P = 0.0001) during intervention whereas no significant changes were observed in the control group. Initial proliferative PBMC responses differed between the vitamin E group and the control group whereas all other baseline characteristics were comparable. No significant changes were observed in cellular immune responsiveness when adjusted for initial values in either the control group or the vitamin E group and, after the trial period, responses in the two groups were not significantly different. Similarly, in the vitamin E group no significant changes were found in levels of IgG and IgA raised against Penicillium or IgG4 raised against egg, milk, or wheat proteins. In the control group small but significant increases in IgG anti-Penicillium (P <0.05) and decreases in IgG4 against milk proteins (P < 0.05) were observed. Thus, the results of this study performed with the relatively low dose of 100 mg dl-α-tocopheryl acetate do not support the claims of a beneficial effect of vitamin E intake on the overall immune responsiveness of elderly subjects.

Recent evidence suggests that plasma lutein is better correlated than either β-carotene or lycopene with its respective carotenoid intake and therefore may be a better marker of vegetable intake than either β-carotene or lycopene. In the study reported in this paper, measurements of plasma carotenes and retinol were made in infants from two villages near Peshawar in the North West Frontier Province, Pakistan, in July and November 1993. The approximate age at the start was 14 months, and 101 boys and ninety girls completed the study. Of the usual plasma carotenes, only lutein was measurable in all samples and was correlated with retinol in both boys (r 0.38, P < 0.0001; r 0.35, P < 0.001) and girls (r 0.21, P = 0.038; r 0.307, P = 0.003) at the two time points respectively. In addition, the change in lutein was even more strongly correlated with the change in retinol in both boys (r 0.453, P <0.0001) and girls (r 0.439, P < 0.0001). In August β-carotene was measurable in approximately 8 % of samples and this increased to 31 % in November but there was no correlation between β-carotene and retinol at any time. There were negligible amounts of lycopene and β-cryptoxanthin in plasma at both time points. The mean concentration of plasma retinol in the infants was 0.66 μmol/l at baseline and 59 % of the infants had retinol concentrations < 0.7 μmol/l. In addition, there were fifteen infants whose levels were below 0.35 μmol/l suggesting that vitamin A status in the population was marginal. Food intake of the infants was not monitored in the present study but breast feeding continues for up to 2 years in this part of Pakistan and most infants would be weaned onto selected foods eaten by the family. The close correlation between plasma lutein and retinol suggests that the increase in retinol over the summer season may be attributable to an increased availability of green vegetables to the families. The source of lutein to the infants is most likely to be the breast milk since such vegetables are unlikely to be given to infants except to suck as a weaning food. The results may indicate the potential usefulness of plasma lutein as a marker of changes in vegetable intake and changes in vitamin A status in Third World infants and children.

Granulated polyamide (PA) was tested for use as an external marker to estimate faecal DM (FDM) excretion of Zebu cattle (Bos indicus). The study was conducted in Mali, using seven and eighteen animals respectively in four field trials and six indoor experiments. Cattle ate fresh or dry pasture vegetation and half the animals were additionally supplemented with crop byproducts. Gelatine capsules containing 35, 40 or 45 g PA were administered orally at 12 h intervals. Estimates of FDM were based on the average marker concentration in faeces and were correlated with the actual excretion measured by total faecal collection. The pre-measurement period required to establish equilibrium for regular marker dosing was determined at 4 d. Except for diets with a N content of less than 9·26 g/kg organic matter, marker recovery averaged 98·1 (se 0·93) % (n 62), and was not influenced by diet composition and the quantity of feed ingested (P > 0·05). Estimates of FDM based on average PA concentrations in faecal samples were correlated to the actual excretion with r 0·98 (n 62; P ≤ 0·001). Since the PA concentration in individual faecal grab-samples is not correlated with either sample mass or sampling time, accurate estimates of FDM require a grab-sampling schedule that covers the 24 h day. However, estimates of FDM were found to be acceptable if calculations are based on the average PA concentration in the sub-total of samples collected during the day or during night respectively (r 0·95, n 29; P ≤ 0·001 in both cases). It is concluded that the use of PA marker is a simple and inexpensive method resulting in reliable estimates of FDM. Since sophisticated analytical procedures are not required to recover PA in faecal samples, the marker is particularly suitable for application in extensive grazing systems and in studies conducted in less-developed countries.

The metabolic effects of a phlorizin-induced drainage of glucose were studied in six lactating ewes with or without peroral alanine drenches in a study of crossover design. Phlorizin gave rise to a small, but significant, elevation of plasma β-hydroxybutyrate. The plasma level of alanine decreased by about 30 % due to the phlorizin injections and alanine was negatively correlated to β-hydroxybutyrate. The plasma level of free fatty acids increased due to phlorizin. Plasma insulin and glucose concentrations were not significantly affected by phlorizin while glucagon level showed a small but significant increase. Peroral alanine drenches to phlorizin-treated ewes gave rise to a transitory elevation of alanine in plasma. The plasma level of free fatty acids was about 40 % lower in phlorizin-treated ewes receiving alanine and β-hydroxybutyrate tended to be lower (P < 0.08). We suggest that β-hydroxybutyrate, apart from its function as an oxidative fuel, might play an important role by limiting glucose oxidation and protein degradation in skeletal muscles during periods of negative energy balance in ruminants. Furthermore, it is suggested that alanine supplementation decreases lipolysis and ketogenesis in lactating ewes.

The present experiments were conducted to investigate influences of dietary methionine and cysteine on metabolic responses to immunological stress induced by Escherichia coli lipopolysaccharide (LPS) injection, and concanavalin A (Con A)-induced mononuclear cell (MNC) proliferation in male broiler chickens. In Expt 1, chicks (12 d of age) were fed on a S amino acid (SAA)-deficient diet (5.6 g SAA/kg diet) or on three kinds of SAA-sufficient diet (9.3 g SAA/kg diet; low-, medium- and high-cysteine diets) which contained 2.8, 4.65 and 6.5 g cysteine/kg diet, respectively. Plasma a-1 acid glycoprotein (AGP) concentration and interleukin (IL)-1-like activity in chicks fed on the SAA-deficient diet were lower following a single injection of LPS than those in chicks fed on the SAA-sufficient diets. At 16 h after LPS injection, plasma Fe and Zn concentrations and body weight were reduced, but AGP concentration and IL-1-like activity in plasma were significantly increased. These changes in body weight, plasma Zn and Fe concentrations following injection of LPS were not affected by dietary methionine:cysteine ratios. Plasma AGP concentration and IL-1-like activity in chicks fed on the high-cysteine diet were, however, greater than those in chicks fed on the other diets following a single injection of LPS. In Expt 2, chicks (7 d of age) were fed on the SAA-sufficient diets as in Expt 1 for 10 d. MNC proliferation in spleen induced by Con A in chicks fed on the high-cysteine diet was greater than that in chicks fed on the low- or medium-cysteine diet. The results suggest that dietary cysteine has an impact on the immune and inflammatory responses.

To study the fate of l-cysteine and amino acid homeostasis in liver after the inhibition of the trans-sulfuration pathway, rats were treated with propargylglycine (PPG). At 4 h after the administration of PPG, liver cystathionase (EC 4.4.1.1) activity was undetectable, l-cystathionine levels were significantly higher, l-cysteine was unchanged and GSH concentration was significantly lower than values found in livers from control rats injected intraperitoneally with 0.15 M-NaCl. The hepatic levels of amino acids that are intermediates of the urea cycle, l-ornithine, l-citrulline and l-arginine and blood urea were significantly greater. Urea excretion was also higher in PPG-treated rats when compared with control rats. These data suggest a stimulation of ureagenesis in PPG-treated rats. The inhibition of γ-cystathionase was reflected in the blood levels of amino acids, because the L-methionine: l-cyst(e)ine ratio was significantly higher in PPG-treated rats than in control rats; blood concentration of cystathionine was also greater. Histological examination of liver and kidney showed no changes in PPG-treated rats when compared with controls. The administration of N-acetylcysteine (NAC) to PPG-treated rats reversed the changes in blood urea and in liver GSH. These data suggest that when liver l-cysteine production was impaired by the blockage of the trans-sulfuration pathway, the concentration of this amino acid was maintained mainly by an increase in protein degradation and by a depletion in GSH concentration that may spare l-cysteine.

Wistar rats fed on either a high-protein or a protein-free diet were examined to determine their pancreatic hydrolase mRNA stabilities in comparison with those of control animals receiving a standard diet. Actinomycin D was used to inhibit transcription and, after isolating the pancreatic RNA, the specific messengers were quantified by performing dot-blot hybridization with cDNA probes. In the rats fed on a high-protein diet, only the half-lives of anionic trypsinogen I and elastase I (EC 3.4.21.36) were affected. Interestingly, when rats were fed on the protein-free diet, most of the hydrolase mRNA half-lives showed changes, except that corresponding to lipase. In these rats, the half-life values of the mRNA coding for anionic trypsinogen I, chymotrypsinogen and procarboxypeptidase B increased, in sharp contrast with those of the amylase and elastase I mRNA, which decreased. These results strongly suggest that the mechanism whereby the biosynthesis of pancreatic hydrolases is regulated, depending on the presence or absence of proteins in the diet, is not unique and provide evidence that the stability of mRNA encoding most, if not all, the hydrolases in pancreatic cells is modulated by the dietary protein content.

Rodents fed on a Mg-deficient (Mg-D) diet develop cardiomyopathic lesions, as well as other types of cardiovascular dysfunction. In the rat, inflammatory cell infiltration of the myocardium begins to occur by week 1, and the lesions develop extensively in the third and fourth weeks on the Mg-D diet. Although the aetiologic mechanisms of Mg-D cardiomyopathy are unknown, we have previously reported that once plasma Mg is markedly reduced, one of the earliest molecular markers of the pathophysiological process is elevation of plasma substance P, calcitonin gene-related peptide and prostaglandin E2, followed by histamine and the inflammatory cytokines (interleukin-1, interleukin-6, and tumor necrosis factor-α). In order to evaluate the potential role of specific circulating inflammatory cell subpopulations in the mechanisms underlying pathophysiological changes observed in Mg-deficiency-induced cardiomyopathy, we analysed these cells by flow cytochemistry. Leucocyte subpopulation pools increased progressively in the Mg-D rats. Elevated circulating levels of neutrophils and lymphocytes appeared to contribute to both the acute (week 1–2) and chronic phases (week 3–4) of the inflammatory responses; monocytes, eosinophils, basophils and large unstained cells which are lymphoid in stained smears, on the other hand, increased significantly in the third and fourth weeks and thus contributed to the chronic inflammatory phase. Changes in the circulating leucocyte subpopulations paralleled the chronological progression of the cardiomyopathic lesions, particularly in weeks 3 and 4. Since a pronounced neutrophilia preceded leucocyte infiltration and deposition within the myocardial tissue, modifications of the microvascular barrier may be a prerequisite for cardiomyopathy in this model of neurogenic inflammation.