Because of the discretionary nature of voluntary food fortification in the European Union, there is a need to monitor fortification practices and consumption of fortified foods in order to assess the efficacy and safety of such additions on an ongoing basis. The present study aimed to investigate the nutritional impact of changes in voluntary fortification practices in adults aged 18–64 years using dietary intake data from two nationally representative cross-sectional food consumption surveys, the North/South Ireland Food Consumption Survey (NSIFCS) (1997–9) and the National Adult Nutrition Survey (NANS) (2008–10). The supply of fortified foods increased between 1997–9 and 2008–10, resulting in a higher proportion of adults consuming fortified foods (from 67 to 82 %) and a greater contribution to mean daily energy intake (from 4·6 to 8·4 %). The overall nutrient profile of fortified foods consumed remained favourable, i.e. higher in starch and dietary fibre and lower in fat and saturated fat, with polyunsaturated fat, sugars and Na in proportion to energy. Women, particularly those of childbearing age, remained the key beneficiaries of voluntary fortification practices in Ireland. Continued voluntary fortification of foods has increased protection against neural tube defect-affected pregnancy by folic acid and maintained the beneficial impact on the adequacy of Fe intake. Increased consumption of fortified foods did not contribute to an increased risk of intakes exceeding the tolerable upper intake level for any micronutrient. Recent increases in voluntary fortification of foods in Ireland have made a favourable nutritional impact on the diets of adults and have not contributed to an increased risk of adverse effects.

Recent rat studies show different tissue distributions of vitamin B12 (B12), administered orally as hydroxo-B12 (HO-B12) (predominant in food) and cyano-B12 (CN-B12) (common in supplements). Here we examine male Wistar rats kept on a low-B12 diet for 4 weeks followed by a 2-week period on diets with HO-B12 (n 9) or CN-B12 (n 9), or maintained on a low-B12 diet (n 9). Plasma B12 was analysed before, during and after the study. The content of B12 and its variants (HO-B12, glutathionyl-B12, CN-B12, 5'-deoxyadenosyl-B12 (ADO-B12), and methyl-B12 (CH3-B12)) were assessed in the tissues at the end of the study. A period of 4 weeks on the low-B12 diet reduced plasma B12 by 58 % (from median 1323 (range 602–1791) to 562 (range 267–865) pmol/l, n 27). After 2 weeks on a high-B12 diet (week 6 v. week 4), plasma B12 increased by 68 % (HO-B12) and 131 % (CN-B12). Total B12 in the tissues accumulated differently: HO-B12>CN-B12 (liver, spleen), HO-B12<CN-B12 (kidneys), and HO-B12≈CN-B12 (brain, heart). Notably, more than half of the administered CN-B12 remained in this form in the kidneys, whereas HO-B12 was largely converted to the bioactive ADO-B12. Only <10 % of the other cofactor, CH3-B12, were found in the tissues. In conclusion, dietary CN-B12 caused a higher increase in plasma and total kidney B12 but provided less than half of the active coenzymes in comparison to dietary HO-B12. These data argue that HO-B12 may provide a better tissue supply of B12 than CN-B12, thereby underscoring the lack of a direct relation between plasma B12 and tissue B12.

The second year of life is a period of nutritional vulnerability. We aimed to investigate the dietary patterns and nutrient intakes from 1 to 2 years of age during the 12-month follow-up period of the Growing Up Milk – Lite (GUMLi) trial. The GUMLi trial was a multi-centre, double-blinded, randomised controlled trial of 160 healthy 1-year-old children in Auckland, New Zealand and Brisbane, Australia. Dietary intakes were collected at baseline, 3, 6, 9 and 12 months post-randomisation, using a validated FFQ. Dietary patterns were identified using principal component analysis of the frequency of food item consumption per d. The effect of the intervention on dietary patterns and intake of eleven nutrients over the duration of the trial were investigated using random effects mixed models. A total of three dietary patterns were identified at baseline: ‘junk/snack foods’, ‘healthy/guideline foods’ and ‘breast milk/formula’. A significant group difference was observed in ‘breast milk/formula’ dietary pattern z scores at 12 months post-randomisation, where those in the GUMLi group loaded more positively on this pattern, suggesting more frequent consumption of breast milk. No difference was seen in the other two dietary patterns. Significant intervention effects were seen on nutrient intake between the GUMLi (intervention) and cows’ milk (control) groups, with lower protein and vitamin B12, and higher Fe, vitamin D, vitamin C and Zn intake in the GUMLi (intervention) group. The consumption of GUMLi did not affect dietary patterns, however, GUMLi participants had lower protein intake and higher Fe, vitamins D and C and Zn intake at 2 years of age.

The molecular basis of the positive association between apoE4 genotype and CVD remains unclear. There is direct in vitro evidence indicating that apoE4 is a poorer antioxidant relative to the apoE3 isoform, with some indirect in vivo evidence also available. Therefore it was hypothesised that apoE4 carriers may benefit from α-tocopherol (α-Toc) supplementation. Targeted replacement mice expressing the human apoE3 and apoE4 were fed with a diet poor (0 mg/kg diet) or rich (200 mg/kg diet) in α-Toc for 12 weeks. Neither apoE genotype nor dietary α-Toc exerted any effects on the antioxidant defence system, including glutathione, catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase activities. In addition, no differences were observed in mitogen-induced lymphocyte proliferation. α-Toc concentrations were modestly higher in plasma and lower in tissues of apoE4 compared with apoE3 mice, with the greatest differences evident in the lung, suggesting that an apoE4 genotype may reduce α-Toc delivery to tissues. A tendency towards increased plasma F2-isoprostanes in apoE4 mice was observed, while liver thiobarbituric acid-reactive substances did not differ between apoE3 and apoE4 mice. In addition, C-reactive protein (CRP) concentrations were reduced in apoE4 mice indicating that this positive effect on CRP may in part negate the increased CVD risk associated with an apoE4 genotype.

1. Pigs, each surgically prepared with a Thiry-Vella jejunal loop were used to study the absorption of calcium and inorganic phosphate from the intestine.

2. The loops were perfused daily for 6–8 h with a nutrient solution and absorption from the perfusate was measured.

3. The technique employed minimized atrophy of the jejunal mucosa and enabled the measurement of hourly or daily changes in absorption rate of components of the luminal fluid.

4. No differences were observed when polyethylene glycol (PEG; molecular weight 4000), [14C]PEG or 51Cr-EDTA were used as markers of net water movement.

5. Increasing the concentration of Ca in the perfusate resulted in the demonstration of a two-component relationship between net absorption rate of Ca and intraluminal Ca concentration. An initial rapid absorption rate from 0 to 4 mm was found, then a slower rate from approximately 5 mm upwards which did not saturate at the highest concentration tested (25 mm).

6. Increasing the concentration of phosphate in the perfusion solution increased the net absorption of phosphate from that solution. Although a two-component system, similar to that for Ca, was not evident, net absorption of phosphate was not saturated at the highest concentration tested (50 mm).

7. The absorption of Ca was unaffected by the presence of phosphate in the solution but Ca (2.5 mm) enhanced the absorption of phosphate.

1. Bipolar electrodes were permanently implanted on the gastric antrum, and on the different portions of the small intestine of each of eleven healthy adult cats receiving one meal daily. All parts of the feline gut exhibited, as in several other species, regular slow waves and alternate periods of quiescence and electrical spiking activity during the recording sessions lasting from 10 to 30 d.

2. Patterns of electrical activity characteristic of this species were identified. Both the amplitude and frequency of the antral slow-wave were related to the presence cf superimposed spike bursts during fasting decrease in the antral slow-wave frequency and increase in the length of the duodenal plateau of slow waves after the daily meal were related to its nature.

3. In fasted state, the electrical spiking activity of the small intestine occurred as fused spike bursts of large amplitude potentials migrating slowly over short distances only 24 h after feeding. They are interspersed with short periods of irregular spiking activity.

4. These findings suggested that, except the distal part of the small intestine which showed an activity which resembled partially the migrating myoelectric complex observed in other species during the fasting state, the motility patterns of the digestive tract in the cat were not comparable to those observed in the dog or sheep. In the cat, mixing of the contents seemed to result from more or less regular spiking activity allowing their propulsion distally. The propagation over distances varying from 200 to 1000 mm of nine to eighteen daily fused spike bursts in the fasting state remains unclear but they are related to the digestive function in accordance with the displacement aborally of their origin in a prolonged fasting condition.

1. The potential use of an extrinsic label to measure iron absorption from a ferric orthophosphate-fortified malted cocoa drink was examined by measuring the solubility of the FePO4 in 0·1 M-hydrochloric acid.

2. The validity of using the stable isotope 58Fe as an extrinsic label was tested by comparing Fe absorption by rats from wheat flour extrinsically-labelled with 58Fe or 59Fe.

3. Fe absorption from a malted cocoa drink (20 g powder made up with hot water) fortified with FePO4 (0·5 mg Fe/g powder) was measured in human subjects using 58Fe as an extrinsic label. Absorption was calculated by measuring unabsorbed 58Fe in faeces. Absorptions of Fe from the drink fortified with either FePO4 or ferrous sulphate were compared. The effect of the addition of ascorbic acid to the drink (1 mg/g powder) on Fe availability was also examined.

4. The effect of fasting on Fe absorption from the drink was determined in rats by giving the drink to fasting animals or shortly after they had consumed a small meal.

5. The FePO4 was totally soluble in 0·1 M-HCl and there were no differences in absorption between 58Fe- and 59Fe-labelled wheat flour. In the human experiment the proportion of Fe absorbed from the drink plus FePO4 and ascorbic acid was (mean with SE) 0·25 (0·02), from the drink plus FePO4 0·24 (0·02) and from the drink plus FeSO4 0·23 (0·03). Fasting had a significant effect on Fe availability; rats given the drink shortly after a small meal absorbed less than half as much Fe as those given the drink on a fasted stomach.

6. It was concluded that the FePO4 used to fortify the malted cocoa drink was as well absorbed as FeSO4 but that the high levels of absorption were a reflection of the fasting condition of the subjects. The level of ascorbic acid was not great enough to enhance the availability of the FePO4 any further.

n-3 Fatty acids may protect against heart disease mortality by preventing fatal arrhythmias. Underlying effects on cardiac electrophysiology may be demonstrable in the standard electrocardiogram (ECG) and provide insight into the mechanism. Therefore, we investigated the effect of dietary n-3 fatty acids on heart-rate-corrected QT interval, T-loop width, spatial QRS-T angle and spatial U-wave amplitude in patients with frequent premature ventricular complexes. Seventy-four patients received either capsules providing 1·5 g n-3 fatty acids daily or placebo for approximately 14 weeks. ECG were recorded before and after intervention. None of the ECG characteristics was significantly affected by treatment. The present results do not provide additional support for the hypothesis that n-3 fatty acids prevent cardiac arrhythmia through generic electrophysiologic effects on heart cell membranes. However, we cannot exclude effects of n-3 fatty acids on clinical relevant endpoints that are not easily detected by prior changes in the ECG.

The aim of the present study was to assess how high doses of dietary vitamin K influence the intestinal profile of K-vitamins in vitamin K-deficient rats, and whether the induced changes are reflected in the hepatic vitamin K store. Vitamin K-deficient rats were fed for 10 d on diets containing different forms of vitamin K, and it was determined how these diets affected the vitamin K concentration at various sites of the instestine, serum, and the liver. It was found that the absorption of phylloquinone from standard food is not more than 10 %, while the absorption of pharmacological doses of oil-solubilized phylloquinone and menaquinone-4 was also far from complete (18 and 55 % respectively). High intakes of phylloquinone suppress the colonic production of all higher menaquinones. High menaquinone-4 intake induces very high menaquinone-8 concentrations, both in the colonic contents as well as in the liver. These data suggest that menaquinone-4 may be converted into menaquinone-8 (but not into other menaquinones) via a metabolic pathway which has not been reported previously.

The present study investigated the relative importance of glutamine as a transamination source in the ovine liver by examination of the labelling of amino acids (AA) in the hepatic free pool, mixed liver and plasma proteins of fed and fasted sheep, following infusion of isotopically-labelled glutamine. In a cross-over design four sheep were either fasted for 3 d or fed to 1·2×energy maintenance and finally euthanased. At each intake, the sheep were infused for 6 h with [2-15N]glutamine (150 μmol/h) and samples of total plasma protein isolated. Following the terminal infusion, liver tissue total proteins were prepared and hydrolysed and 15N-enrichments in seventeen AA were determined by GC–combustion–isotope-ratio mass spectrometry. All AA were enriched (relative to natural abundance) except lysine and threonine, with the lowest enrichments in phenylalanine and histidine. There was no effect of the fed v. fasted state, except for leucine and isoleucine in liver protein (P<0·05). Enrichments in liver protein were greater than in plasma protein (P<0·01, except proline) and probably reflect the faster turnover rate of hepatic constitutive proteins compared with export proteins. Amination to methionine was greater than that to phenylalanine (P<0·01), suggesting a mechanism for preferentially protecting the former. This factor could be important for ruminant production, as methionine is often considered to be the first limiting AA for animals offered certain silages and conserved forages. Enrichments in all AA (except for glutamine, alanine and aspartate) were less than that for glutamate (P<0·01), and thus transaminations may have occurred with glutamine directly or via glutamate, following the action of hepatic glutaminase.

Diabetes-induced CVD is the most significant complication of prolonged hyperglycaemia. The aim of this study was to determine whether resveratrol, a polyphenol antioxidant compound, when administered at a dose that can be reasonably obtained through supplementation could prevent the development of cardiovascular complications in older, obese, diabetic rats. Diabetes was induced in 6-month old, obese, male Wistar rats via a single intravenous dose of streptozotocin (65 mg/kg). Randomly selected animals were administered resveratrol (2 mg/kg) via oral gavage daily for 8 weeks. Body weights, blood glucose levels, food intake and water consumption were monitored, and assessments of vascular reactivity, tactile allodynia and left ventricular function were performed. Resveratrol therapy significantly improved tactile allodynia and vascular contractile functionality in diabetic rats (P<0·05). There were no significant changes in standardised vasorelaxation responses, plasma glucose concentrations, water consumption, body weight, left ventricular hypertrophy, kidney hypertrophy, heart rate or left ventricular compliance with resveratrol administration. Resveratrol-mediated improvements in vascular and nerve function in old, obese, diabetic rats were associated with its reported antioxidant effects. Resveratrol did not improve cardiac function nor mitigate the classic clinical symptoms of diabetes mellitus (i.e. hyperglycaemia, polydypsia and a failure to thrive). This suggests that supplementation with resveratrol at a dose achievable with commercially available supplements would not produce significant cardioprotective effects in people with diabetes mellitus.

Hypertension is a key feature of the metabolic syndrome. Lifestyle and dietary changes may affect blood pressure (BP), but the knowledge of the effects of dietary fat modification in subjects with the metabolic syndrome is limited. The objective of the present study was to investigate the effect of an isoenergetic change in the quantity and quality of dietary fat on BP in subjects with the metabolic syndrome. In a 12-week European multi-centre, parallel, randomised controlled dietary intervention trial (LIPGENE), 486 subjects were assigned to one of the four diets distinct in fat quantity and quality: two high-fat diets rich in saturated fat or monounsaturated fat and two low-fat, high-complex carbohydrate diets with or without 1·2 g/d of very long-chain n-3 PUFA supplementation. There were no overall differences in systolic BP (SBP), diastolic BP or pulse pressure (PP) between the dietary groups after the intervention. The high-fat diet rich in saturated fat had minor unfavourable effects on SBP and PP in males.

Low dietary K levels have been associated with increasing renal Ca excretion in humans, indicating a higher risk of calcium oxalate (CaOx) urolith formation. Therefore, the present study aimed to investigate whether dietary K also affects the urine composition of cats. A total of eight adult cats were fed diets containing 0·31 % native K and 0·50, 0·75 and 1·00 % K from KCl or KHCO3 and were evaluated for the effects of dietary K. High dietary K levels were found to elevate urinary K concentrations (P< 0·001). Renal Ca excretion was higher in cats fed the KCl diets than in those fed the KHCO3 diets (P= 0·026), while urinary oxalate concentrations were generally lower in cats fed the KCl diets and only dependent on dietary K levels in cats fed the KHCO3 diets (P< 0·05). Fasting urine pH increased with higher dietary K levels (P= 0·022), reaching values of 6·38 (1·00 % KCl) and 7·65 (1·00 % KHCO3). K retention was markedly negative after feeding the cats with the basal diet ( − 197 mg/d) and the 0·50 % KCl diet ( − 131 mg/d), while the cats tended to maintain their balance on being fed the highest-KCl diet ( − 23·3 mg/d). In contrast, K from KHCO3 was more efficiently retained (P= 0·018), with K retention being between − 82·5 and 52·5 mg/d. In conclusion, the dietary inclusion of KHCO3 instead of KCl as K source could be beneficial for the prevention of CaOx urolith formation in cats, since there is an association between a lower renal Ca excretion and a generally higher urine pH. The utilisation of K is distinctly influenced by the K salt, which may be especially practically relevant when using diets with low K levels.

Perinatal maternal high-fat diet (HFD) increases susceptibility to obesity and fatty liver diseases in adult offspring, which can be attenuated by the potent hypolipidaemic action of fish oil (FO), an n-3 PUFA source, during adult life. Previously, we described that adolescent HFD offspring showed resistance to FO hypolipidaemic effects, although FO promoted hepatic molecular changes suggestive of reduced lipid accumulation. Here, we investigated whether this FO intervention only during the adolescence period could affect offspring metabolism in adulthood. Then, female Wistar rats received isoenergetic, standard (STD: 9 % fat) or high-fat (HFD: 28·6 % fat) diet before mating, and throughout pregnancy and lactation. After weaning, male offspring received the standard diet; and from 25 to 45 d old they received oral administration of soyabean oil or FO. At 150 d old, serum and hepatic metabolic parameters were evaluated. Maternal HFD adult offspring showed increased body weight, visceral adiposity, hyperleptinaemia and decreased hepatic pSTAT3/STAT3 ratio, suggestive of hepatic leptin resistance. FO intake only during the adolescence period reduced visceral adiposity and serum leptin, regardless of maternal diet. Maternal HFD promoted dyslipidaemia and hepatic TAG accumulation, which was correlated with reduced hepatic carnitine palmitoyl transferase-1a content, suggesting lipid oxidation impairment. FO intake did not change serum lipids; however, it restored hepatic TAG content and hepatic markers of lipid oxidation to STD offspring levels. Therefore, we concluded that FO intake exclusively during adolescence programmed STD offspring and reprogrammed HFD offspring male rats to a healthier metabolic phenotype in adult life, reducing visceral adiposity, serum leptin and hepatic TAG content in offspring adulthood.

In a previous trial in Mali, we showed that traditional pearl millet couscous and thick porridge delayed gastric emptying (about 5 h half-emptying times) in a normal-weight population compared with non-traditional carbohydrate-based foods (pasta, potatoes, white rice; about 3 h half-emptying times), and in a gastric simulator we showed millet couscous had slower digestion than wheat couscous. In light of these findings, we tested the hypothesis in a normal-weight US population (n 14) that millet foods would reduce glycaemic response (continuous glucose monitor), improve appetitive sensations (visual analogue scale ratings), as well as reduce gastric emptying rate (13C-octanoic acid breath test). Five carbohydrate-based foods (millet couscous – commercial and self-made, millet thick porridge, wheat couscous, white rice) were fed in a crossover trial matched on available carbohydrate basis. Significantly lower overall glycaemic response was observed for all millet-based foods and wheat couscous compared with white rice (P ≤ 0·05). Millet couscous (self-made) had significantly higher glycaemic response than millet couscous (commercial) and wheat couscous (P < 0·0001), but as there were no differences in peak glucose values an extended glycaemic response was indicated for self-made couscous. Millet couscous (self-made) had significantly lower hunger ratings and higher fullness ratings (P < 0·05) than white rice, millet thick porridge and millet couscous (commercial). A normal gastric emptying rate (<3 h half-emptying times) was observed for all foods, with no significant differences among them. In conclusion, some traditionally prepared pearl millet foods show the potential to reduce glycaemic response and promote satiety.