Following an introduction of the importance of folates and the rationale for seeking to estimate fractional folate absorption from foods (especially for countries not having a mandatory folic acid fortification policy), scientific papers covering the mechanisms of folate absorption and initial biotransformation are discussed. There appears (post-1983) to be a consensus that physiological doses of folic acid undergo biotransformation in the absorptive cells of the upper small intestine to 5-methyltetrahydrofolic acid (as happens for all naturally-occurring reduced 1-carbon-substituted folates). This ‘validates’ short-term experimental protocols assessing ‘relative’ folate absorption in human subjects that use folic acid as the ‘reference’ dose. The underlying scientific premise on which this consensus is based is challenged on three grounds: (i) the apparent absence of a 5-methyltetrahydrofolic acid response in the human hepatic portal vein following absorption of folic acid, (ii) the low dihydrofolate reductase activity peculiar to man and (iii) the implications derived from recent stable-isotope studies of folate absorption. It is concluded that the historically accepted case for folic acid being a suitable ‘reference folate’ for studies of the ‘relative absorption’ of reduced folates in human subjects is invalid. It is hypothesised that the liver, and not the absorptive cells of the upper small intestine, is the initial site of folic acid metabolism in man and that this may have important implications for its use as a supplement or fortificant since human liver's low capacity for reduction may eventually give rise to saturation, resulting in significant (and potentially deleterious) unmetabolised folic acid entering the systemic circulation.

Cu is an essential nutrient for man, but can be toxic if intakes are too high. In sensitive populations, marginal over- or under-exposure can have detrimental effects. Malnourished children, the elderly, and pregnant or lactating females may be susceptible for Cu deficiency. Cu status and exposure in the population can currently not be easily measured, as neither plasma Cu nor plasma cuproenzymes reflect Cu status precisely. Some blood markers (such as ceruloplasmin) indicate severe Cu depletion, but do not inversely respond to Cu excess, and are not suitable to indicate marginal states. A biomarker of Cu is needed that is sensitive to small changes in Cu status, and that responds to Cu excess as well as deficiency. Such a marker will aid in monitoring Cu status in large populations, and will help to avoid chronic health effects (for example, liver damage in chronic toxicity, osteoporosis, loss of collagen stability, or increased susceptibility to infections in deficiency). The advent of high-throughput technologies has enabled us to screen for potential biomarkers in the whole proteome of a cell, not excluding markers that have no direct link to Cu. Further, this screening allows us to search for a whole group of proteins that, in combination, reflect Cu status. The present review emphasises the need to find sensitive biomarkers for Cu, examines potential markers of Cu status already available, and discusses methods to identify a novel suite of biomarkers.

Fe-deficiency anaemia is the most common cause of anaemia in developing countries. In these settings, many chronic infections, including tuberculosis (TB), are highly prevalent. Fe is an essential nutrient for both host and mycobacteria that play a pivotal role in host immunity and mycobacterial growth. A case–control study was performed in a TB-endemic region in Jakarta, Indonesia, among 378 pulmonary TB patients and 436 healthy controls from the same neighbourhood with the same socio-economic status. In a number of these subjects the Fe status could be explored. The distribution of three polymorphisms in the natural resistance-associated macrophage protein gene (NRAMP1) including INT4, D543N and 3′UTR was examined for a possible association with susceptibility to TB. Anaemia (corrected for sex) was present in 63·2 % of active TB compared with 6·8 % of controls, with female patients more often affected. Anaemia was more pronounced in advanced TB as diagnosed by chest radiography. Lower Hb concentrations in TB patients were accompanied by lower plasma Fe concentrations, lower Fe-binding capacity and higher plasma ferritin. After successful TB therapy, Fe parameters improved towards control values and Hb levels normalised, even without Fe supplementation. NRAMP1 gene polymorphisms were not associated with TB susceptibility, TB severity or anaemia. In conclusion, most active TB patients had anaemia, which was probably due to inflammation and not to Fe deficiency since TB treatment without Fe supplementation was sufficient to restore Hb concentration.

Plant extracts rich in polyphenols (PERP) could represent interesting alternative antioxidants but their use in ruminants needs further investigation since the antioxidant capacity of PERP could be altered by digestive processes. The aim of the study was to investigate the bioavailability and the antioxidant capacity of four PERP (rosemary; grape; citrus; marigold) in ruminants made highly susceptible to lipoperoxidation by a continuous linseed oil infusion (4 % DM) in the duodenum. The PERP were given, as a single acute dose (10 % DM), directly into the rumen of sheep (n 5) and blood was then collected every 3 h over a period of 30 h. Grape was particularly efficient to enhance the plasma total antioxidant status (P < 0·05). Moreover, many new polyphenols were detected in the plasma and the identification of epicatechin in the grape group suggested that, contrary to monogastrics, ruminants can benefit from the antioxidant effect of polymeric proanthocyanidins. Finally, the four PERP tested, and more especially marigold, significantly reduced plasma susceptibility to liperoxidation (mean increase of lag phase: +5·9 min, P < 0·02; mean reduction of oxidation rate: − 1·7 A234/min, P < 0·01). In conclusion, the digestive processes in ruminants do not inhibit the antioxidant properties of PERP in vivo and are beneficial by improving the biological effect of polymeric proanthocyanidins. Further experiments are now necessary to determine the optimum dose of administration and to characterize the bioactive molecules.

Miglitol, a 1-deoxynojirimycin derivative, is an α-glucosidase inhibitor. In the present study, the effects of acute (single-dose) and chronic (8-week) oral administration of miglitol in Goto-Kakizaki (GK) rats, an animal model of type 2 diabetes, were investigated. Dose-dependent decreases in incremental blood glucose concentrations integrated over a period of 2 h (ΔAUC0–2 h) for values of blood glucose after sucrose-loading in miglitol-treated GK rats were observed following an acute oral administration of miglitol (1, 3 or 10 mg/kg body weight). At 10 mg/kg, the ΔAUC0–2 h of blood glucose was decreased by 45 % compared with the control group. Following the oral administration of miglitol in a dietary mixture (10 mg, 20 mg or 40 mg miglitol/100 g control diet) for 8 weeks, the ratio of HbA1c at 8 weeks compared with 0 weeks in GK rats treated with 40 mg miglitol/100 g control diet miglitol was significantly decreased compared with control GK rats without changes in body weight. In oral glucose tolerance testing, miglitol caused a slight decrease in the ΔAUC0–2 h of plasma glucose concentration. In addition, miglitol treatment slightly inhibited the reduction in β-cell mass, and lessened the irregular contours and fibrosis of the islets in GK rats. These results indicate that miglitol ameliorates the hyperglycaemic state of GK rats and the impaired function of the pancreatic islets, as well as preventing the degeneration of islets in GK rats.

Taurine has been shown to be tissue protective in many models of oxidant-induced injury. However, its protective role against retinal damage induced by photochemical stress is less well known. The purpose of the present study was to investigate whether dietary taurine reduced retinal photochemical damage in Sprague–Dawley rats and to further explore the underlying molecular mechanisms of this action. Twenty rats fed AIN-93 formulation and maintained in the dark for 48 h were used as controls (n 20). Another forty rats were randomly divided into two groups and then treated with (n 20) or without 4 % taurine (n 20) for 15 d respectively. After treatment, these two groups were exposed to fluorescent light (3000 ± 200 lux and 25°C), and the protective effects of dietary taurine were then evaluated. The present results showed that dietary taurine effectively prevented retinal photochemical damage as assessed by changes of morphology. Also, the supplementation caused an increase of taurine in the retina, a decrease of malondialdehyde (P < 0·01), and elevation of superoxide dismutase (P < 0·01) and glutathione peroxidase activities in the retina (P < 0·01). Moreover, dietary taurine inhibited activator protein-1 (AP-1) (c-fos/c-jun subunits) expression (P < 0·05), up regulated NF-κB (p65) expression (P < 0·05), and decreased caspase-1 expression (P < 0·05) so as to reduce the apoptosis of photoreceptors in the retina (P < 0·05). These results suggest that dietary taurine reduced retinal damage produced by photochemical stress via antioxidant and anti-AP-1–NF-κB–caspase-1 apoptotic mechanisms in rats.

The improved effects of dietary chickpeas on visceral adiposity, dyslipidaemia and insulin resistance were examined. Rats were fed a normal-fat diet (NFD), a high-fat diet (HFD) or a high-fat plus chickpea diet (HFD+CP) for 8 months. The epididymal fat pad weight v. total body weight of rats was higher in the HFD group (0·032 (sd 0·0042) g/g) than in the NFD group (0·015 (sd 0·0064) g/g) and smaller in the HFD+CP group (0·023 (sd 0·0072) g/g) compared with the HFD group (P < 0·05). Chickpea treatment also induced a favourable plasma lipid profile reflecting decreased TAG, LDL-cholesterol (LDL-C) and LDL-C:HDL-cholesterol levels (P < 0·05). HFD-fed rats had higher TAG concentration in muscle and liver, whereas the addition of chickpeas to the HFD drastically lowered TAG concentration (muscle, 39 %; liver, 23 %). The activities of lipoprotein lipase (LPL) in epididymal adipose tissue and hepatic TAG lipase in liver recorded a 40 and 23 % increase respectively in HFD rats compared with those in NFD rats; dietary chickpeas completely normalised the levels. Furthermore, chickpea-treated obese rats also showed a markedly lower leptin and LPL mRNA content in epididymal adipose tissue. An insulin tolerance test, oral glucose tolerance test and insulin-releasing test showed that chickpeas significantly improved insulin resistance, and prevented postprandial hyperglycaemia and hyperinsulinaemia induced by the chronic HFD. The present findings provide a rational basis for the consumption of chickpeas as a functional food ingredient, which may be beneficial for correcting dyslipidaemia and preventing diabetes.

Gallic acid (GA) is a naturally abundant plant phenolic compound in the human diet and is known to reduce the risk of disease. In this study, the anti-obesity effect of GA in an animal model of diet-induced obesity was investigated. Obesity was induced in male Wistar rats by feeding them a high-fat diet (HFD). GA was given as a supplement at the levels of 50 and 100 mg/kg rat for a period of 10 weeks. The results showed that the body weight, organ weight of the liver and adipose tissue weights of peritoneal and epididymal tissues in the HFD+GA groups were significantly decreased as compared with the HFD group. Serum TAG, phospholipid, total cholesterol, LDL-cholesterol, insulin and leptin levels in the HFD+GA groups were significantly decreased as compared with the HFD group. Histological study showed that the lipid droplets of rats with HFD+GA diets were significantly smaller than those with HFD diets. Hepatic TAG and cholesterol levels in HFD+GA groups were significantly decreased as compared with the HFD group. Moreover, the consumption of GA reduced oxidative stress and GSSG content and enhanced the levels of glutathione, GSH peroxidase, GSH reductase and GSH S-transferase in the hepatic tissue of rats with HFD-induced obesity. These results demonstrate that intake of GA can be beneficial for the suppression of HFD-induced dyslipidaemia, hepatosteatosis and oxidative stress in rats.

The aim of this study was to evaluate the effect of a synbiotic containing Lactobacillus acidophilus ATCC 4962, fructooligosaccharide, inulin and mannitol on plasma lipid profiles and erythrocyte membrane properties in hypercholesterolaemic pigs on high- and low-fat diets. Twenty-four white male Landrace pigs were randomly allocated to four treatment groups for 8 weeks (n 6). Treatment factors were the supplementation of synbiotic (with and without) and dietary fat (5 and 15 %). The supplementation of synbiotic reduced plasma total cholesterol (P = 0·001), TAG (P = 0·002) and LDL-cholesterol (P = 0·045) for both dietary fats. A higher concentration of esterified-cholesterol in HDL of pigs supplemented with synbiotic than the control regardless of dietary fat (P = 0·036) indicated that cholesterol was reduced in the form of cholesteryl esters. Reduced concentration of cholesteryl esters (P < 0·001) and increased concentration of TAG (P = 0·042) in LDL of pigs on synbiotic suggested that LDL-cholesterol was reduced via the hydrolysis of smaller and denser LDL particles. The erythrocytes of pigs without any synbiotic showed more prevalence of spur cells than those given the synbiotic, as supported by the higher cholesterol: phospholipid ratio in erythrocytes (P = 0·001). Also, membrane fluidity and rigidity were improved as supported by the decreased fluorescence anisotropies in the Hb-free erythrocyte membrane of pigs given synbiotic (P < 0·001). The administration of the synbiotic reduced plasma TAG, total cholesterol and LDL-cholesterol in hypercholesterolaemic pigs, possibly in the form of cholesteryl esters, via the interrelated pathways of lipid transporters (VLDL, LDL and HDL). The synbiotic also reduced deformation of erythrocytes via improved membrane fluidity and permeability.

The effects of phytic acid and microbial phytase on the flow and composition of endogenous protein at the terminal ileum of broiler chickens were investigated using the peptide alimentation method. Phytic acid (fed as the sodium salt) was included in a synthetic diet at 8·5, 11·5 and 14·5 g/kg (or 2·4, 3·2 and 4·0 g/kg phytate-phosphorus) and each diet was fed without or with an Escherichia coli-derived microbial phytase at 500 phytase units/kg diet. A control containing no phytate was fed as a comparison to estimate basal endogenous flows. Ingestion of phytic acid increased (P < 0·05) the flow of endogenous amino acids and N by an average of 47 % at the lowest phytic acid concentration and 87 % at the highest. The addition of microbial phytase reduced (P < 0·05) the inimical effects of phytic acid on endogenous amino acid flow at all dietary phytic acid levels. The composition of endogenous protein was also influenced (P < 0·10–0·001) by increasing phytic acid concentrations and phytase addition. The effects of phytic acid and phytase on endogenous flow and composition of endogenous protein, however, varied depending on the amino acid. It is concluded that the effects of phytase on amino acid digestibility may be mediated, in part, through a route of reduced endogenous loss.

Two incubation runs were conducted with Rusitec fermenters to investigate the effects of three additive treatments (mixed fibrolytic enzymes from Trichoderma longibrachiatum (FE), disodium fumarate (FUM) and both additives (MIX)) on rumen microbial growth and fermentation of a grass hay:concentrate (600 : 400 g/kg DM) substrate. Each fermenter received daily 20 g substrate DM. Application rate (per g substrate DM) was 34·3 endoglucanase, 0·57 exoglucanase, 24·7 xylanase and 5·51 amylase units for FE and 30 mg fumarate for FUM. MIX fermenters received both additives. Both FE and MIX increased (P < 0·05) daily production of acetate, butyrate and methane, substrate DM and fibre disappearance at 6 and 48 h incubation, daily flow of microbial-N, and the microbial colonisation of substrate at 6 h incubation. Compared to FE, MIX treatment increased (P < 0·05) propionate production by 28 % and decreased (P < 0·05) the acetate:propionate ratio, but no other differences between both treatments were found (P>0·05). Supplementing with FUM increased (P < 0·05) volatile fatty acid production by 11 % and decreased (P < 0·05) the acetate:propionate ratio, but did not affect (P>0·05) any other variable, thus suggesting that observed effects were due to fermentation of FUM itself. The lack of effects of FUM and the absence of differences between FE and MIX on most of the measured variables would indicate that beneficial effects found in MIX fermenters were mainly due to the action of FE. Combining FE and FUM as feed additives under the conditions of the present experiment did not further improve rumen fermentation, compared to FE alone.

The present study evaluated levels of growth factors and their associations with nutritional status with emphasis on stunting in children at 1 and 3 years of age. A follow-up study on a birth cohort (n 219) of children from villages in the central region of the Limpopo Province was undertaken. Of the original cohort, 156 and 162 could be traced and assessed at ages 1 and 3 years, respectively. Data collected included socio-demographic characteristics, anthropometric measurements, dietary intake and fasting blood (collected from 116 and 145 children at 1 and 3 years, respectively) for growth factor analysis (insulin-like growth factor (IGF)-1, IGF binding protein (BP)-1, IGFBP-3, leptin, glucose and insulin). At 1 year it was found that stunted children had lower leptin levels while their IGFBP-1 levels were higher than that in normal children. These differences were, however, not observed at 3 years. Furthermore at 1 year the biochemical parameters were more related to length measures whereas at 3 years the parameters were more associated with weight measures. The observed stunting in this group of children may be a result of chronic undernutrition resulting in long-term growth faltering which is already evident at 1 year. Thus the observed phenomenon might be an adaptive mechanism adopted by children's metabolic processes as they grow up in an environment with inadequate essential nutrients due to poor weaning practices and consumption of a diet of poor quality, resulting in them gaining more weight at the expense of linear growth.

Within a decade or so insufficient fish oil (FO) will be available to meet the requirements for aquaculture growth. Consequently, alternative sources are being investigated to reduce reliance on wild fish as a source of FO. Vegetable oils (VO) are a feasible alternative to FO. However, it is important to establish that alternative dietary lipids are not only supplied in the correct quantities and balance for optimal growth, but can maintain immune function and prevent infection, since it is known that the nutritional state of the fish can influence their immune function and disease resistance. A way of maintaining immune function, while replacing dietary FO, is by using a blend of VO rather than a single oil. In this study, juvenile European sea bass (Dicentrarchus labrax) were fed diets with a 60 % substitution of FO with a blend of rapeseed, linseed and palm oils. Two oil blends were used to achieve a fatty acid composition similar to FO, in terms of energy content, and provide a similar balance of SFA, MUFA and PUFA. Fish were fed the diets for 64 weeks, after which time growth and fatty acid compositions of liver and blood leucocytes were monitored. The impact of the dietary blends on selected innate immune responses and histopathology were also assessed, together with levels of plasma prostaglandin E2. The results suggest that potential exists for replacing FO with a VO blend in farmed sea bass feeds without compromising growth, non-specific immune function or histology.

Food-derived bioactive peptides are reported to express a variety of functions in vivo. We studied the in vitro effect of three bioactive tripeptides, isoleucine-proline-proline (IPP), valine-proline-proline (VPP) and leucine-lysine-proline (LKP), on osteoblast proliferation and gene expression. We used UMR-106 osteosarcoma cells, human marrow-derived mesenchymal stem cells (hMSC) and osteoblasts differentiated from hMSC. Treatment with 50 μm-IPP increased UMR-106 cell and hMSC proliferation. The gene expression of hMSC-differentiated osteoblasts was analysed by the microarray method. Microarray analysis revealed that IPP up-regulated 270 genes and down-regulated 100 genes. VPP and LKP, by contrast, had a very modest influence on osteoblast gene expression. Real-time PCR confirmed that IPP up-regulated PTHrP, BMP-5 and CREB-5 and down-regulated VDR and caspase-8. IPP possesses potential to increase osteoblast proliferation, differentiation and signalling. Agents that increase the number and function of osteoblasts could improve bone mass and structure, and decrease fracture risk.

In the present study, we utilised an in vitro digestion procedure to deliver molecules contained in tomatoes to cultured cells and to analyse potential mechanisms underlying the antitumoural effects of tomatoes reported in the literature. Ripe tomatoes underwent in vitro simulated digestion and the aqueous fraction obtained was delivered to HT-29 and HCT-116 colon adenocarcinoma cells. The amount of lycopene released during digestion and transferred to the aqueous fraction during digestion was 10-fold lower than that present in tomato homogenate before digestion. The carotenoid was accumulated by colon adenocarcinoma cells in a dose-dependent manner after the addition of tomato digestate (20–100 ml/l) for 24 h. Tomato digestate inhibited the growth of HT-29 and HCT-116 cells in a dose-dependent manner. Growth inhibition resulted from an arrest of cell cycle progression at the G0/G1 phase and by apoptosis induction. A down regulation of cyclin D1, Bcl-2 and Bcl-xL expression was also observed, without apparent changes in p53, p21, p27 and Bax. In conclusion, the present data demonstrate that the in vitro digestion procedure represents a useful approach to supply tomato to colon cultured cells. Moreover, we have shown that tomato digestate is able to inhibit the growth of colon cancer cells by modulating the expression of regulators of the cell cycle and apoptosis.

Children with history of broncho-pulmonary dysplasia (BPD) often suffer from growth failure and lung sequelae. The main objective of this study was to test the role of pulmonary obstruction on resting energy expenditure (REE) and nutritional status in BPD. Seventy-one children with BPD (34 boys and 37 girls) and 30 controls (20 boys and 10 girls) aged 4–8 years were enrolled. Body composition was assessed by bio-impedancemetry measurements; REE was measured by indirect calorimetry. Predicted REE was calculated using the Schofield equation. The population of children with BPD was divided into three groups: children without obstruction of the airways, children with moderate obstruction of the airways, and children with severe obstruction. Children with BPD were significantly smaller and leaner than controls. Altered body composition (reduction of fat mass) was observed in BPD children that suffered from airway obstruction. REE was significantly lower in children with BPD compared to controls, but when adjusted for weight and fat-free mass no significant difference was observed irrespective of pulmonary status. Airway obstruction in children with BPD does not appear to be associated with an increased REE. Moreover altered REE could not explain the altered nutritional status that is still observed in BPD in later childhood. This supports the hypothesis that body composition and pulmonary function in BPD in later childhood are fixed sequelae originating from the neonatal period.

Salicylic acid (SA) is a natural phenolic compound known as the active principle of aspirin. Its presence in vegetal sources suggests that fruit and vegetable (FV) consumption could produce measurable SA serum concentrations in human subjects not taking aspirin. The aim of this study was to investigate the relationship between FV intake and circulating SA in healthy subjects. Thirty-eight volunteers (twenty-two males and sixteen females) were recruited from an Italian university campus. They recorded their food intake for 7 d to evaluate dietary consumption and, in particular, FV intake; fasting blood samples were taken on the morning of the eighth day to measure SA serum concentration, using a sensitive stable isotope dilution and GC–MS method. Median SA serum concentration was 0·124 μmol/l (range 0·028–0·295). Circulating SA was significantly related to FV consumption, both to the mean daily intake (r2 0·13, P = 0·03) and to the last day intake (r2 0·16, P = 0·01). The subjects in the highest FV intake quartile in the preceding day (>4·75 servings) had significantly higher SA concentrations than in the lowest quartile ( < 2·3 servings) (median concentrations 0·188 and 0·112 μmol/l, respectively; P = 0·04). This study proved that, after overnight fast, human subjects not taking aspirin display circulating SA in amounts related to the FV consumption. It is therefore possible that the beneficial effects of regular FV consumption in man could also depend on low chronic SA exposure.

Postmenopausal women with the metabolic syndrome are at high risk of oxidative stress. Several studies have suggested possible antioxidant properties of soya, but little evidence is available regarding the effect of soya on oxidative stress in postmenopausal women with the metabolic syndrome. The objective of the present study was to determine the effects of soya consumption on plasma total antioxidant capacity (TAC) and malondialdehyde (MDA) level in postmenopausal women with the metabolic syndrome. A randomised cross-over trial was undertaken on forty-two postmenopausal women with the metabolic syndrome. Participants were randomly assigned to consume a control (Dietary Approaches to Stop Hypertension; DASH) diet, a soya protein diet, or a soya nut diet, each for 8 weeks. Red meat in the DASH diet (one serving per d) was replaced by soya protein in the soya protein period and by soya nuts in the soya nut period. Significant differences between the end values of the control diet, soya protein diet and soya nut diet were seen for MDA (0·70, 0·64 and 0·63 μmol/l; global P < 0·01). The results also showed a significant difference between the end values for TAC (1950, 2030 and 2110 μmol/l, respectively; P < 0·01). The difference from control for TAC was +4·5 % (P < 0·01) in the soya nut period and +5·8 % (P < 0·01) in the soya protein regimen. Both soya nuts and soya protein decreased MDA significantly compared with the control diet (difference from control was − 7·9 % (P < 0·01) in the soya nut period and − 9·4 % (P < 0·01) in the soya protein diet). We conclude that soya consumption reduces plasma MDA and increases plasma TAC levels in postmenopausal women with the metabolic syndrome.