Zn deficiency has been shown to reduce host defence drastically. It was of interest to determine the capacity of the residual lymphocytes from Zn-deficient mice to proliferate and produce lymphokines in response to stimulation since there are many Zn-dependent metalloenzymes that might be altered by the deficiency. To address this question, young adult A/J mice were provided Zn-deficient or Zn-adequate diets or restricted amounts of a Zn-adequate diet for 30 d. Splenocytes from moderately or severely Zn- deficient adult A/J mice gave normal proliferative responses and generated adequate interleukin II (IL- 2) activity when stimulated with the mitogen Concanavalin A. However, splenocytes from deficient mice exhibited a higher degree of proliferation (about 150%) and production of IL-2 in response to foreign target cells compared with T-cells prepared from mice provided a Zn-adequate diet. B-cells from deficient mice stimulated in vivo with sheep erythrocytes produced fewer total numbers of plaque-forming cells (PFC) per spleen. Nevertheless, the proportion or number of PFC/106 viable splenocytes and the amounts of IgM and IgG antibody produced per PFC were equivalent to those of adequately-fed and restricted-fed controls. The previously described responses were not significantly affected by whether the level of Zn in the culture medium was adequate or limiting. Based on these tests it appeared that the residual splenic lymphocytes of Zn-deficient mice were able to carry out many fundamental immune processes.

A continuous in vitro method for the estimation of the bioavailability of minerals and trace elements is presented. This in vitro method is believed to be more representative of in vivo physiological conditions than in vitro methods based on equilibrium dialysis, because dialysable components are continuously removed from the pancreatic digestion mixture. The continuous in vitro method is compared with the equilibrium in vitro method with respect to the dialysability of Ca, Mg, Fe, Cu and Zn from eight different types of bread (varying in phytic acid content). The results show a pronounced effect of continuous removal of dialysable components from the pancreatic digestion mixture on the dialysability of minerals and trace elements. Furthermore, removal of dialysable components influences the effect of phytic acid on the bioavailability of minerals and trace elements. For these two reasons the importance of removal of dialysable components in vitro for the estimation of bioavailahility in vivo needs further investigation. The bioavailability of minerals and trace elements from bread samples is not related to the phytic acid content only. Therefore, the effect of phytic acid on the bioavailability of minerals and trace elements cannot be studied separately from the effects of other components on bioavailability.

The effect of lactose on the urinary excretion of Mg and Ca, as an index of absorption, was studied in a double-blind, crossover study during three 1-week periods. Twenty-four healthy, lactose-tolerant, adult volunteers maintained their habitual diets with the exception that all lactose-containing dairy products in the diet were replaced by 600 g/d of three specially prepared dairy products. These products were based on either lactose-enriched cow's milk or lactose-enriched, lactase (EC 3.2.1.23)-treated cow's milk, with or without added Mg, and were given in turn during 1 week. Lactose intake was increased by 127 mmol/d (46 g/d) while taking the lactose-enriched products. While taking the Mg-enriched products, Mg intake was increased by 2.8 mmol/d (69 mg/d) which was equivalent to 17% of the habitual Mg intake. Apart from the lactose and Mg intake, nutrient intake was comparable during the three dietary periods. Urinary excretions of Mg and Ca were used as indicators for their absorption. Mg supplementation significantly increased urinary Mg excretion by 0.97 mmol/d (equivalent to an increase of 18%, P < 0.001), indicating that urinary Mg excretion is a valid indicator for intestinal Mg absorption. Hydrolysis of lactose did not affect urinary excretion of Mg and Ca, which implies that lactose intake does not affect the absorption of Mg and Ca in healthy adults.

An increase in dietary intake of B from 0.25 to 3.25 mg/d has been reported to increase plasma oestradiol and testosterone and decrease urinary Ca excretion in postmenopausal women. Consequently, it is suggested that the higher level of B intake could reduce bone loss associated with the menopause and cessation of ovarian function. The present study was designed to investigate the effect of a B supplement on bone mineral absorption and excretion, plasma sex steroid levels and urinary excretion of pyridinium crosslink markers of bone turnover in healthy postmenopausal volunteers. The women were accommodated in a metabolic unit, given a low-B diet (LBD; 0.33 mg/d) for 3 weeks and were asked to take a B supplement of 3 mg/d in addition to the LBD for a further 3 weeks. Changing B intake from 0.33 to 3.33 mg/d had no effect on minerals, steroids or crosslinks. However, the LBD appeared to induce hyperabsorption of Ca since positive Ca balances were found in combination with elevated urinary Ca excretion. This phenomenon may have inhibited or obscured any effect of B.

The Ca and P intakes of 148 pregnant and lactating women in a rural village in The Gambia. West Africa, have been estimated by direct weighing of food on a total of 4188 d. The Ca and P contents of local foods were determined by analysis of raw ingredients, snack foods and prepared dishes. Information about the contribution of mineral-rich seasonings was obtained. Efforts were made to discover unusual sources of Ca that might not be perceived as food by subject or observer. The main contributors to daily Ca intake were shown to be leaves, fish, cereals, groundnuts and local salt. Cow's milk accounted for only 5 % of Ca intake. Unusual sources of Ca were discovered, namely baobab (Adansonia digitata) fruit and selected earths, but these were consumed infrequently and their contributions to Ca intakes were small. Cereals and groundnuts were the main sources of P. Ca and P intakes (mg/d) were shown to average 404 (SD 110) and 887 (SD 219) respectively. Seasonal changes in the availability of leaves, cereals and groundnuts resulted in variations in Ca and P intakes. The rainy season was associated with increased Ca intakes (by 16%) but decreased P consumption (by 15%). No difference was observed in Ca intake between pregnant and lactating women but P intake in lactation was 11 % higher than that in pregnancy during the post-harvest season. The implications of these low Ca intakes require investigation.

Xanthan gum (15 g/d) was given for 10 d to eighteen normal volunteers. In vivo measurements of stool output, transit time, frequency of defaecation and flatulence were compared with a preceding control period of 10 d. At the end of the control and test periods fresh faecal homogenate from each subject was anaerobically incubated with xanthan gum and control solutions to assess the ability of the bacteria to break down the gum. Xanthan gum was found to be a highly efficient laxative agent causing significant increases in stool output (P < 0.01), frequency of defaecation (P < 0.05) and flatulence (P < 0.01) whilst having variable effects on transit time. Before feeding xanthan gum, faecal samples from twelve of the eighteen subjects could reduce the viscosity of the gum in vitro. This rose to sixteen of the eighteen with significantly greater amounts (P < 0.05) of hydrogen and short-chain fatty acids also being produced, indicating bacterial adaptation in the presence of the substrate. Correlations between the in vivo and in vitro findings did not substantiate claims that the in vivo effect of a given polysaccharide can be predicted from its fermentation characteristics in vitro.

The purpose of the present paper was to study the effects of a dietary undigestible carbohydrate and intestinal microflora on mucin distribution (neutral, acid, sulphonated), glycolytic activities: η-D-galactosidase (EC 3.2.1.23), N-acetyl-η-D-galactosaminidase ( Ec 3.2.1.43), N-acetyl-η-D-gluco-saminidase (EC 3.2.1.30), β-L-fucosidase (EC 3.2.1.51) and bacterial metabolism (gas production, short-chain fatty acids (SCFA) and lactic acid caecal concentration) in germ-free (GF), conventional (CV) and heteroxenic (HE) rats (GF rats associated with a human flora). Rats were fed on either a control diet or a diet containing 40 g trans-galactosylated oligosaccharide (TOS)/kg. In GF rats fed on the control diet caecal pH was almost neutral and glycolytic activities negligible. The number of mucus- containing cells increased from the caecum to the colon for the three types of mucin. TOS had no effect in the caecum but it modified mucin cell repartition in the colon. In CV and HE rats fed on the control diet caecal pH was similar (6.8), but caecal SCFA and lactic acid concentrations (μmol/g) and gas production (m1/24 h) were higher in CV (70, 5.9 and 2.3 respectively) than in HE rats (32, 4.6 and 0.4 respectively). In CV, as in HE rats, acid-mucin-containing cells increased from the caecum to the colon and glycolytic activities were similar. TOS reduced acid-mucin-containing cells in the caecum of CV rats by twofold but had no effect in either the caecum or the colon of HE rats. TOS strongly increased η- galactosidase activity and slightly modified the other glycolytic activities. Its effect on bacterial metabolites depended on bacterial status. However, comparison between CV and HE rats showed no evident relationship between the number of mucus-containing cells and measured bacterial metabolites. Differences between CV and HE rats might be due to bacterial microflora specificity. TOS had an intrinsic effect on mucus cell distribution in the colon of GF rats. In CV and HE rats the presence of the flora abolished this effect.

Wheat bran and pectin (100 g/kg) were added to a basal diet and fed to rats. An in vitro dialysis technique was used to measure the distribution of caecal and faecal water between the bound, i.e. that held by bacteria and undigested macromolecules, and free water. Bran increased wet (67%) and dry (74%) faecal weight. Pectin increased wet faecal weight (59 %), but did not influence dry weight. In faeces both bran and pectin increased the amount of total and bound water, but only pectin increased total and bound water when expressed on a dry weight basis. Caecal wet (90%) and dry (67%) weights increased with pectin but not with bran. Bran did not change total water but increased bound water whereas pectin increased both. This suggests that water contributed more to the increase in stool bulk in the pectin- supplemented animals due to free and bound water associated with both increased numbers of bacteria and residual pectin. Pectin altered the distribution of water in faeces. Bran has no effect on water distribution and is only partly fermented. The residual water-holding capacity leads to an increased wet and dry stool output.

Studies were carried out to determine the effects of the toxic principle linamarin, a cyanogenic glucoside, in a diet containing cassava (Manihot esculenta Crantz) in the form of gari fed to growing dogs for 14 weeks. There were three groups of dogs, each comprising six animals. One group was fed on a control diet with rice as the carbohydrate source, the second group was fed on cassava (gari) as the carbohydrate source and which was expected to release 10.8 mg HCN/kg cooked food, the third group was fed on the control diet to which enough NaCN was added at feeding time to release 10.8 mg HCN/kg cooked food in order to monitor the effects of the HCN released from gari. All diets contained 130 g erude protein (N × 6.25)/kg and were supplemented with vitamins and minerals. Each animal was given approximately 100 g diet/kg body weight for the duration of the experifnent. The biochemical variables investigated were plasma electrolytes, serum proteins, plasma-free amino acids, plasma enzymes and urine protein, and the histology of some metabolically active tissues, namely liver, kidney, myocardium, testis and adrenal gland, was studied. The gari diet caused an elevated plasma thiocyanate concentration (P < 0.01), elevated 24 h urinary thiocyanate excretion and elevated urinary protein excretion (P < 0.01), lowered serum albumin (P < 0.05), a plasma-free amino acid profile which resembled that found in kwashiorkor, lowered plasma K and Ca (P < 0.05). The rice + cyanide diet caused an elevated plasma thiocyanate (P < 0.01) and a 24 h urinary thiocyanate excretion that was significantly higher (P < 0.01) than that of the dogs fed on gari, but caused a urinary protein excretion that was significantly lower than that of the dogs fed on gari (P < 0.01), lowered serum albumin (P < 0.05), a plasma-free amino acid profile that indicated that the amino acids were not being utilized to the same extent as in the control (rice) group but were accumulating. Neither diet had an effect on plasma γ-glutamyltransferase (EC 2.3.2.2), alanine aminotransferase (EC2.6.1.2) or isocitrate dehydrogenase (EC 1.1.1.42) activities, plasma Na, Mg, and P concentrations. The gari diet caused generalized congestion and haemorrhage, periportal vacuolation of the liver, swelling, vacuolation and rupture of the epithelial cells of the proximal convoluted tubules of the kidney, myocardial degeneration and adrenal gland degeneration. In the testes there were occasional abnormal germ cells in the seminiferous tubules, and occasional seminiferous tubules denuded to basement membrane of germ cells but with remnants of Sertoli cells. Spermatogenesis, however, appeared to be normal since the percentage of round tubules in stage 8 of the spermatogenic cycle was not significantly different from that of the control dogs. The rice + cyanide diet caused nephrosis and a significantly reduced relative frequency of testicular tubules in stage 8 of the spermatogenic cycle (P < 0.01). There was also marked testicular germ cell sloughing and degeneration. There was adrenal gland hyperplasia and hypertrophy. It was concluded that the observed changes which occurred when the gari diet was consumed were not entirely due to cyanide.