Chronic inflammation of the vascular endothelium produces endothelial dysfunction and ultimately atherogenesis. Postprandial hyperlipidaemia is an independent risk factor for cardiovascular disease. Recent studies show that the magnitude of postprandial lipaemia following a single fatty meal is negatively related to vascular function. This is associated with a transient increase in the concentrations of pro-inflammatory cytokines and soluble adhesion molecules and in pro-oxidant activity. One possible interpretation is that repeated exposure of the blood vessel wall to the activities of pro-inflammatory cytokines and pro-oxidants may damage the vascular endothelium and promote atherogenesis. Based on these results, we propose a model of a causal mechanism to explain how consumption of a fatty meal may impair vascular dysfunction.

Alkylresorcinols (AR), a group of phenolic lipids, exist in the human diet in whole-grain rye and wheat. They are absorbed by humans and have been quantified in plasma. In this 2-week study we assessed AR incorporation into human erythrocyte membranes. Nine subjects attended the study; four avoided whole-grain products for 1 week and then included whole-grain rye and wheat bread in the diet for the second week, four included whole-grain rye and wheat products in the diet during the whole follow-up and one followed a gluten-free diet. Plasma and erythrocyte membrane AR were analysed after weeks 1 and 2. Erythrocyte membrane AR concentrations increased an average of 231 nmol/l of packed erythrocytes (P=0·036) after consumption of whole-grain rye and wheat products. Plasma AR levels increased an average of 175 nmol/l (P=0·058) When intake of whole-grain products was constant, erythrocyte membrane and plasma AR levels remained stable. Long-chain AR were incorporated into erythrocyte membranes in a higher proportion compared to shorter-chain AR. This preliminary study shows that AR are incorporated into human erythrocyte membranes in vivo.

Recent studies have shown an inverse association between the level of dietary Ca, particularly from dairy sources, and body weight in adults; there is, however, a paucity of data regarding this relationship in children. We therefore investigated this issue in 1087 children who underwent body weight and height measurement during a survey on childhood obesity. Lifestyle and dietary habits were investigated by a questionnaire. After excluding children who were following a dietary regimen for any reason, 884 children (M:F 451:433; age 7·5 (sd 2·1) years) were selected. Milk consumption was pooled into four frequency categories: poor (≤1/week; n 125), moderate (>1 but ≤5–6/week; n 133), regular (1/d; n 408) and high (≥2/d; n 218). The frequency of consumption of milk was inversely and significantly associated (t=–2·64, P=0·003) with age- and sex-specific BMI z-scores by linear regression analysis, controlling for sex, age, physical activity, birth weight and parental overweight and education. The statistical association remained significant (t=–2·831, P=0·005) after the inclusion of children consuming only skimmed milk (n 91). Milk consumption was still significantly and inversely associated with BMI z score (t=–2·791, P=0·005) in the whole-milk consumers when controlling for age and the frequency of consumption of various foods; this association was no longer significant (P=0·21) when children consuming skimmed milk were included in the analysis. This is the first report showing a significant inverse association between frequency of milk consumption and body mass in children. Regardless of the mechanisms involved, our results might encourage further research on this issue and might have important implications for the prevention of obesity.

Si-Jun-Zi decoction (SJZD), a traditional Chinese herbal prescription, has been used clinically for treating patients with disorders of the digestive system. Previous studies indicated that the polysaccharides of SJZD (SJZPS) are the active components contributing towards its pharmacological effects in improving gastrointestinal function and immunity. However, the protective and restitutive effects on intestinal epithelial cells remain unknown. In the present study, SJZPS were first extracted and chemically characterized. Then their stimulatory and restitutive effects on intestinal epithelial cells (IEC-6 cells) were elicited by different in vitro models including migration of wounded IEC-6 cells and cell proliferation. Results indicated that SJZPS not only protects the cells against the harmful impairment of indomethacin but also enhances re-epithelialization of a wounded monolayer at an optimal dose of 100 μg/ml at 24 h incubation. To elucidate the modulatory effect of SJZPS on wounded IEC-6 cells at the molecular level, an oligonucleotide microarray was employed to study differential gene expression of SJZPS-treated IEC-6 cells and the candidate genes were validated by RT-PCR. There was increased expression of genes coding for ion channels and transporters, which are critical to cell migration and restoration of wounded intestinal cells, suggesting a possible mechanism for re-epithelialization. In conclusion, our data show for the first time that SJZPS can enhance intestinal restitution and protect against indomethacin-induced damage of intestinal epithelial cells. These findings provide new insight into the mechanism of action of a traditional Chinese herbal prescription, SJZD, in intestinal wound restitution.

The availability of folate is implicated as a determinant of DNA methylation, a functionally important feature of DNA. Nevertheless, when this phenomenon has been examined in the rodent model, the effect has not always been observed. Several reasons have been postulated for the inconsistency between studies: the rodent is less dependent on folate as a methyl source than man; juvenile animals, which most studies use, are more resistant to folate depletion than old animals; methods to measure genomic DNA methylation might not be sensitive enough to detect differences. We therefore examined the relationship between folate and genomic DNA methylation in an elder rat model with a newly developed method that can measure genomic DNA methylation sensitively and precisely. Thirty-nine 1-year-old rats were divided into three groups and fed a diet containing 0, 4·5 or 18 μmol folate/kg (folate-deplete, -replete and -supplemented groups, respectively). Rats were killed at 8 and 20 weeks. At both time points, mean liver folate concentrations increased incrementally between the folate-deplete, -replete and -supplemented rats (P for trend <0·001) and by 20 weeks hepatic DNA methylation also increased incrementally between the folate-deplete, -replete and -supplemented rats (P for trend=0·025). At both time points folate-supplemented rats had significantly increased levels of DNA methylation compared with folate-deplete\ rats (P<0·05). There was a strong correlation between hepatic folate concentration and genomic DNA methylation in the liver (r 0·48, P=0·004). In the liver of this animal model, dietary folate over a wide range of intakes modulates genomic DNA methylation.

It is well known that early weaning causes marked changes in intestinal structure and function, and transforming growth factor-β (TGF-β) is believed to play an important regulatory role in post-weaning adaptation of the small intestine. The present study examined the distribution and expression intensity of TGF-β in the small intestinal mucosa of pre- and post-weaning pigs using a specific immunostaining technique and Western blot analysis. The level of TGF-β in the intestinal mucosa, as estimated by Western blot analysis, did not change significantly during weaning. However, when examined by the immunostaining technique, TGF-β1 (one of the TGF-β isoforms dominantly expressed in the tissue) at the intestinal villus epithelium, particularly at the apical membrane of the epithelium, decreased significantly 4 d after weaning, while the staining intensity increased significantly at the intestinal crypts compared with that in pre-weaning pigs. These changes were transient, with the immunostaining intensity for TGF-β1 at the intestinal villi and the crypts returning to the pre-weaning level by 8 d post-weaning. The transient decrease in TGF-β1 level at the intestinal villus epithelium was associated with obvious intestinal villus atrophy and marked reduction of mucosal digestive enzyme activities. Furthermore, the number of leucocytes staining positively for TGF-β1 increased significantly in the pig intestinal lamina propria 4 d after weaning. These findings strongly suggest that TGF-β plays an important role in the post-weaning adaptation process in the intestine of the pig.

Protein malnutrition can adversely affect all tissues. The aim of the present study was to test the hypothesis that protein deprivation influences gastric ulcer formation, as well as metabolism and organ growth, in rats. In the present study, there was a significant reduction in the body and organ weight of rats fed a low-protein diet (P<0·001). Malnourished rats were less susceptible to ulceration of the gastric mucosa in ethanol and indomethacin models of acute gastric ulcers when compared with rats fed a normoproteic diet (17 % protein). Mucus production and prostaglandin E2 formation increased in malnourished rats, possibly explaining the lower number of acute ulcers in these animals. Pylorus ligature altered gastric juice composition (increased pH and gastric volume, and decreased total acid concentration) in the animal group fed a low-protein diet compared with the group fed a diet containing 17 % protein (P<0·05). The gastric mucosa was more damaged in malnourished rats than in normal rats evaluated for 14 d after acetic acid injection (P<0·001). Malnourished rats exhibited resistance to acute gastric lesions, owing to an increase in prostaglandin GE2 release and mucus secretion, which protected their gastric mucosa. This phenomenon was not seen in subchronic gastric ulceration.

To investigate exogenous triacylglycerol turnover, the time courses for labelled triolein in the liver, plasma and epididymal adipose tissue (adipose tissue) after oral administration to rats fed a fat-free or 10 % corn oil diet for 3 d after fasting overnight were examined for 10 d. After the administration of labelled triolein to rats fed the fat-free diet, the incorporation (dpm/g) into total lipids of the liver and adipose tissue each reached the maximum in 8 h and was seven times higher in the adipose tissue than in the liver. The half-lives of total lipid radioactivities during the decreasing phases were 0·39 and 2·58 d, respectively, in the rapid and slow phases of the decay curve in the liver, and 4·78 d in only one phase of the adipose tissue. Radioactivity after administration of labelled triolein was mostly found in the oleic acid in the tissues. The half-life of oleic acid was 3·92 d in the adipose tissues. These half-lives were similar in both dietary groups. Thus, although dietary corn oil reduced the triolein incorporation to cellular lipids in comparison to the fat-free diet, it did not affect these half-lives. The labelled triacylglycerol–oleic acid stayed abundantly intact for a long time in the adipose tissue and was scarcely changed to other fatty acids, whereas it was slightly incorporated into total lipids and quickly metabolized in the liver. Non-essential fatty acids may be mostly endogenous in the liver but may be exogenous and endogenous in adipose tissue.

Vitamin A (retinol) is essential for epithelial cell growth, differentiation and proliferation. The absorption of retinol occurs in the small intestine, and the metabolism of this vitamin is not well studied in this organ. The intestinal epithelium has a high rate of cell proliferation and differentiation, and the present study looked at the level of retinoids and metabolizing enzymes involved in their interconversion along the villus–crypt axis under normal conditions. Intestine was removed from control rats, and enterocytes at various stages of maturation and differentiation were quantified by the metal chelation method. Using HPLC, various retinoid concentrations in the cell homogenate and the metabolizing enzymes in the cytosol were quantified. The proliferating crypt cells were found to have a higher level of retinoic acid as well as of the enzymes involved in its formation, such as retinaldehyde oxidase and retinol dehydrogenase, compared with the villus cells, suggesting a possible role for this compound in intestinal epithelial cell proliferation and differentiation. The high level of retinol and high retinaldehyde reductase activity in the villus cells suggest the important role played by this enzyme in the conversion of dietary β-carotene to retinol via retinaldehyde. In summary, this study has given for the first time a detailed analysis of the retinoid levels and metabolizing enzymes in different cell populations in the rat small intestinal epithelium.

In this three-phase study we first compared the availability of fourteen Fe forms in a wheat-based ready-to-eat breakfast cereal using an in vitro digestion/human colonic adenocarcinoma (CaCo-2) cell model. We then investigated the effect of milk and/or coffee on those fortified cereals found in phase 1 to show promising increases in Fe availability. The Fe forms assessed in phase 1 were reduced (control), carbonyl, electrolytic, FePO4, FeSO4, FeCO3, Na2FeEDTA, Ferrochel® (Albion Laboratories, Clearfield, UT, USA; ferrous bis-glycinate), encapsulated ferrous fumarate, FeSO4, ferrous lactate and Biofer® (LipoTech, Britwell Salome, Oxfordshire; FeSO4), SQM® (Sea-Questra-Min Iron; Quali Tech, Chaska, MN, USA; polysaccharide-complexed FeSO4) and Sun Active® (Taiyo Kagaku, Yokkaichi, Japan). All these forms increased Fe uptake compared with the unfortified cereal. Relative to the control, the following increases in Fe availability were observed: electrolytic, 52 %; ferrous fumarate, 30–35 %; Sun Active, 78 %; Ferrochel, 125 %; Na2FeEDTA, 291 %. Recent human studies have shown similar data with regard to Ferrochel, FeSO4 and Na2FeEDTA, with the latter being more bioavailable. Our phase-2 studies indicated that the addition of milk to FeSO4-fortified cereal increased Fe availability, but this availability was markedly decreased by the addition of coffee to the digest. Conversely, a loss in availability from Na2FeEDTA was observed with the addition of milk; however, the addition of coffee did not markedly affect Fe availability from this form. In phase-3 studies we observed increased Fe availability upon the addition of milk to cereals containing Ferrochel, FeSO4, Sun Active, a mixture of reduced Fe and Na2FeEDTA or reduced Fe. For these forms we did not assess the behaviour after the addition of coffee. In conclusion, when considering possible fortificants for optimizing Fe bioavailability within a foodstuff, it is of paramount importance to consider the interaction between the fortified foodstuffs and other components of the meal (such as milk and coffee with a breakfast).

The present work aimed to differentiate between proteolytic activities of plants and micro-organisms during the incubation of grass in cattle rumens. Freshly cut ryegrass was placed in bags of varying permeability and incubated for 16 h in the rumens of dairy cows that had previously grazed a ryegrass sward, supplemented with 4 kg dairy concentrate daily. Woven polyester bags (50 μm pore size) permitted direct access of the micro-organisms and rumen fluid enzymes to the plant material. The polythene was impermeable even to small molecules such as NH3. Dialysis tubing excluded micro-organisms and rumen enzymes/metabolites larger than 10 kDa. DM loss was 46·3 % in polyester, 36·2 % in polythene and 38·1 % in dialysis treatments. It is possible that the DM loss within polythene bags occurred due to a solubilisation of plant constituents (e.g. water-soluble carbohydrates) rather than microbial attachment/degradation processes. The final protein content of the herbage residues was not significantly different between treatments. Regardless of bag permeability, over 97 % of the initial protein content was lost during incubations in situ. Electrophoretic separation showed that Rubisco was extensively degraded in herbage residues whereas the membrane-associated, light-harvesting protein remained relatively undegraded. Protease activity was detected in herbage residues and bathing liquids after all incubation in situ treatments. Although rumen fluid contains proteases (possibly of plant and microbial origin), our results suggest that, owing to cell compartmentation, their activity against the proteins of intact plant cells is limited, supporting the view that plant proteases are involved in the degradation of proteins in freshly ingested herbage.

In rats rendered hypercholesterolaemic by maintaining them on a cholesterol-enriched diet (0·5 %) for 8 weeks, as a result of alteration in membrane structural lipids, erythrocytes were observed to be deformed and become more fragile. This deformity and fragility was partially reversed by the two dietary spice principles, curcumin and capsaicin, and the spice, garlic, by virtue of their ability to lower the extent of hypercholesterolaemia. A further insight into the factors that might have reduced the fluidity of erythrocytes in hypercholesterolaemic rats revealed changes in fatty acid profile of the membranes, phospholipid composition of the membrane bilayer, reduced Ca2+, Mg2+-ATPase, and reduction in the sensitivity of erythrocytes to concanavaline A. Dietary capsaicin appeared to counter these changes partially in hypercholesterolaemic rats. Electron spin resonance (ESR) spectra and fluorescence anisotropy parameters also revealed altered fluidity of erythrocytes in hypercholesterolaemic rats. Dietary capsaicin and curcumin significantly reversed this alteration. Scanning electron microscopic examination revealed that the echinocyte population was increased in the erythrocytes of hypercholesterolaemic rats, and this was significantly countered by dietary capsaicin. The membrane protein profile and the active cation efflux appeared to be unaffected in the hypercholesterolaemic situation.

The salutary characteristics of the tomato are normally related to its content of carotenoids, especially lycopene, and other antioxidants. Our purpose was to verify whether the daily intake of a beverage prototype called Lyc-o-Mato® containing a natural tomato extract (Lyc-o-Mato® oleoresin 6 %) was able to modify plasma and lymphocyte carotenoid concentrations, particularly those of lycopene, phytoene, phytofluene and β-carotene, and to evaluate whether this intake was sufficient to improve protection against DNA damage in lymphocytes. In a double-blind, cross-over study, twenty-six healthy subjects consumed 250 ml of the drink daily, providing about 6 mg lycopene, 4 mg phytoene, 3 mg phytofluene, 1 mg β-carotene and 1·8 mg α-tocopherol, or a placebo drink. Treatments were separated by a wash-out period. Plasma and lymphocyte carotenoid and α-tocopherol concentrations were determined by HPLC, and DNA damage by the comet assay. After 26 d of consumption of the drink, plasma carotenoid levels increased significantly: concentrations of lycopene were 1·7-fold higher (P<0·0001); of phytofluene were 1·6-fold higher (P<0·0001); of phytoene were doubled (P<0·0005); of β-carotene were 1·3-fold higher (P<0·05). Lymphocyte carotenoid concentrations also increased significantly: that of lycopene doubled (P<0·001); that of phytofluene was 1·8-fold higher (P<0·005); that of phytoene was 2·6-fold higher (P<0·005); that of β-carotene was 1·5-fold higher (P<0·01) In contrast, the α-tocopherol concentration remained nearly constant. The intake of the tomato drink significantly reduced (by about 42 %) DNA damage (P<0·0001) in lymphocytes subjected to oxidative stress. In conclusion, the present study supports the fact that a low intake of carotenoids from tomato products improves cell antioxidant protection.

This paper deals with BMI and morbidity in relation to body-fat mass (BFM) and fat-free mass (FFM). The analysis was based on cross-sectional data concerning the age, household income, anthropometry and morbidity of 575 males aged 18–59 years from a rural community in North-East India. Data on morbidity were based on the self-reported morbidity (SRM) of the subjects during the last 4 weeks before the survey, whereas data on BMI and body composition were estimated from anthropometry. It was found that SRM was significantly associated with age and income. However, the relationship between BMI and SRM was not significant after adjusting for age and income. Separating the BMI into body-fat mass index (BFMI being BFM in kg divided by height squared in metres) and fat-free mass index (FFMI being FFM in kg divided by height squared in metres), it was found that BFMI was significantly associated with SRM after adjusting for age, income and FFMI. The subjects with a low (<2·9 kg/m2) BFMI were about 4·7 times (odds ratio 4·7, 95 % CI 2·6, 8·6) more likely to become sick than those with a normal (2·9–5·0 kg/m2) BFMI. In addition, the risk of becoming sick was higher in the subjects with a high (>5·0 kg/m2) BFMI than in those with a normal BFMI (odds ratio 3·9, 95 % CI 1·3, 9·8). However, the relationship between FFMI and morbidity was not clearly perceptible. It is therefore speculated that BMI may not always provide accurate information about the variation in body fat and body composition that is associated with morbidity.

The aim of the present study was to assess the respective contributions of regional and socio-economic factors to dietary pattern. We used the data from the final MONICA (MONItoring of trends and determinants in Cardiovascular disease) population survey conducted in the three French centres in 1995–7 among a representative sample of 976 men aged 45–64 years. Dietary intake was assessed using a 3-d record method. Dietary patterns were identified by a factor analysis, based on fifteen food items. An analysis of variance was then used to study their relationship with regional and socio-economic determinants. Two major dietary patterns were identified: a ‘Western diet’, characterized by high intakes of sugar and sweets, grains, butter, added fats, eggs, potatoes and cheese; a ‘prudent diet’, mainly distinguished by high intakes of fruit, vegetables, olive oil and fish and low intakes of alcohol, high-fat meat and potatoes. Strong associations were mostly observed with the ‘prudent diet’ pattern, with a significant relationship with region, educational and income-tax levels, leisure-time physical activity and smoking status. There was also a statistically significant interaction between region and educational level (P=0·05), and between region and income-tax level (P=0·03), indicating that the influence of socio-economic factors is different among regions. In conclusion, these results indicate large regional and socio-economic differences in the dietary patterns of this French male population. When considering the ‘prudent diet’ pattern, they also suggest that traditional regional influences may now be overcome by socio-economic determinants.

The relative contents of the fatty acids 14: 0, 15: 0 and 17: 0 in serum and adipose tissue may be used as biological markers of dairy fat intake. However, the determinants of these fatty acids are not fully understood. This study investigates the relationship between these fatty acids and the intake of macronutrients and physical activity in a cross-sectional study of 301 healthy men aged 61–64 years. Dietary intake was assessed using a pre-coded 7 d food record, and physical activity during the previous year was recorded in an interview. Under-reporters of energy intake were identified by the Goldberg cut-off. Fatty acid composition was determined in serum phospholipids (PL) and subcutaneous adipose tissue (AT) from the upper buttock. The relative content of each of 14: 0, 15: 0 and 17: 0 in PL and AT was positively associated with the intake of dairy fat. In addition, all three fatty acids were inversely correlated with alcohol intake, R ranging from −0·28 to −0·53 (P<0·001). The results were not markedly affected when under-reporters (n 88) were excluded from the analyses. In both PL and AT, the relative content of the fatty acids was approximately 5% higher in a group of high physical activity compared with a group of low physical activity, although significant trends were only seen for 14: 0 in PL and 17: 0 in AT. The findings suggest that adjustments should be made for alcohol intake when the fatty acids 14: 0, 15: 0 and 17: 0 are applied as markers for dairy fat intake.

Age-related macular degeneration (AMD) is a common disorder that causes irreversible loss of central vision. Increased intake of foods containing zeaxanthin may be effective in preventing AMD because the macula accumulates zeaxanthin and lutein, oxygenated carotenoids with antioxidant and blue light-absorbing properties. Lycium barbarum L. is a small red berry known as Fructus lycii and wolfberry in the West, and Kei Tze and Gou Qi Zi in Asia. Wolfberry is rich in zeaxanthin dipalmitate, and is valued in Chinese culture for being good for vision. The aim of this study, which was a single-blinded, placebo-controlled, human intervention trial of parallel design, was to provide data on how fasting plasma zeaxanthin concentration changes as a result of dietary supplementation with whole wolfberries. Fasting blood was collected from healthy, consenting subjects; fourteen subjects took 15 g/d wolfberry (estimated to contain almost 3 mg zeaxanthin) for 28 d. Repeat fasting blood was collected on day 29. Age- and sex-matched controls (n 13) took no wolfberry. Responses in the two groups were compared using the Mann–Whitney test. After supplementation, plasma zeaxanthin increased 2·5-fold: mean values on day 1 and 29 were 0·038 (sem 0·003) and 0·096 (sem 0·009) μmol/l (P<0·01), respectively, for the supplementation group; and 0·038 (sem 0·003) and 0·043 (sem 0·003) μmol/l (P>0·05), respectively, for the control group. This human supplementation trial shows that zeaxanthin in whole wolfberries is bioavailable and that intake of a modest daily amount markedly increases fasting plasma zeaxanthin levels. These new data will support further study of dietary strategies to maintain macular pigment density.

In this study, we attempted to evaluate the effect of administration of a high quantity of red yeast rice on coenzyme Q10 (CoQ10) synthesis in the tissues of ICR mice. Eighty-eight adult male ICR mice were housed and divided into control and experimental groups for red yeast rice treatment. Animals were gavaged with a low (1 g/kg body weight) or a high dose (5 g/kg body weight, approximately five times the typical recommended human dose) of red yeast rice dissolved in soyabean oil. After gavagement, animals of the control group were immediately killed; mice of the experimental groups (eight for each subgroup) were killed at different time intervals of 0·5, 1, 1·5, 4 and 24 h. The liver, heart and kidney were taken for analysis of monacolin K (liver only) and CoQ10 analysis. Liver and heart CoQ10 levels declined dramatically in both groups administered red yeast rice, especially in the high-dose group, within 30 min. After 24 h, the levels of hepatic and cardiac CoQ10 were still reduced. A similar trend was also observed in the heart, but the inhibitory effect began after 90 min. The higher dose of red yeast rice presented a greater suppressive effect than did the lower dose on tissue CoQ10 levels. In conclusion, acute red yeast rice gavage suppressed hepatic and cardiac CoQ10 levels in rodents; furthermore, the inhibitory effect was responsive to the doses administered.

Techniques have been developed to produce microbial phytase for addition to diets for simple-stomached animals, with the aim to improve phosphorus availability from phytate-P in plant sources. The activityof the crude microbial phytase showed pH optima at pH 5-5 and 2·5. The enzyme was able to degradephytate in vitro in soya-bean meal, maize and a liquid compound feed for pigs. When microbial phytasewas added to low-P diets for broilers the availability of P increased to over 60 % and the amount of Pin the droppings decreased by 50%. The growth rate and feed conversion ratio on the low-P dietscontaining microbial phytase were comparable to or even better than those obtained on control diets.Addition of microbial phytase to diets for growing pigs increased the apparent absorbability of P by24%. The amount of P in the faeces was 35% lower.