An increased activity of ATP-binding cassette transporter (ABC) A1 has been proposed as a mechanism underlying the hypocholesterolaemic effect of phytosterols. In the present study, ABCA1-deficient mice (ABCA1−/− mice) were used to examine the involvement of the ABCA1 in the reduction of intestinal cholesterol absorption in response to a phytosterol-enriched diet. A decrease in intestinal cholesterol absorption of 39 and 35% was observed after phytosterol treatment in ABCA1+/+ mice and in ABCA1−/− mice, respectively. No statistically significant changes in plasma lipoprotein profile or in intestinal ABCG5, ABCG8 and Niemann-Pick C1-Like 1 gene expression levels were found when phytosterol-treated ABCA1−/− mice and untreated ABCA1−/− mice were compared. We conclude that phytosterol inhibition of cholesterol absorption in mice is independent of ABCA1

Dietary cis-9, trans-11-conjugated linoleic acid (CLA) is generally thought to be beneficial for human health. Fish oil added to ruminant diets increases the CLA concentration of milk and meat, an increase thought to arise from alterations in ruminal biohydrogenation of unsaturated fatty acids. To investigate the mechanism for this effect, in vitro incubations were carried out with ruminal digesta and the main biohydrogenating ruminal bacterium, Butyrivibrio fibrisolvens. Linoleic acid (LA) or α-linolenic acid (LNA) was incubated (1·67g/l) with strained ruminal digesta from sheep receiving a 50:50 grass hay–concentrate ration. Adding fish oil (up to 4·17g/l) tended to decrease the initial rate of LA (P=0·025) and LNA (P=0·137) disappearance, decreased (P<0·05) the transient accumulation of conjugated isomers of both fatty acids, and increased (P<0·05) the accumulation of trans-11-18:1. Concentrations of EPA (20:5 n-3) or DHA (22:6 n-3), the major fatty acids in fish oil, were low (100mg/l or less) after incubation of fish oil with ruminal digesta. Addition of EPA or DHA (50mg/l) to pure cultures inhibited the growth and isomerase activity of B. fibrisolvens, while fish oil had no effect. In contrast, similar concentrations of EPA and DHA had no effect on biohydrogenation of LA by mixed digesta, while the addition of LA prevented metabolism of EPA and DHA. Neither EPA nor DHA was metabolised by B. fibrisolvens in pure culture. Thus, fish oil inhibits ruminal biohydrogenation by a mechanism which can be interpreted partly, but not entirely, in terms of its effects on B. fibrisolvens.

UC (ulcerative colitis) patients have an increased risk of developing colorectal cancer compared with the normal population. The cause underlying this higher risk is not fully defined but includes nutritional and environmental factors concomitant with genetic alterations. We aimed to evaluate genetic stability in the colonic tissue of UC patients in clinical remission compared with the healthy population, and to establish a possible correlation between nutritional habits and these molecular assessments. UC patients (n 42) and healthy controls (n 37) participated in the study. All participants were histopathologically and medically diagnosed. Participants completed five separate 7d dietary records, food-frequency questionnaires and validated 24h recalls for nutritional assessment. The extent of chromosome 17 loss and the calculated chromosome index was determined in colon tissue biopsies by fluorescence in situ hybridisation. Correlations between the molecular and nutritional assessments were performed using Pearson's correlation coefficients. Significant differences in the nutritional intake of total fat (65 (SD 15) v. 89 (SD 25) g), cholesterol (330 (SD 168) v. 464 (SD 177) mg), dietary fibre (32 (SD 4·7) v. 9 (SD 4) g), vitamin A (1009 (SD 209) v. 506 (SD 204) μg), vitamin C (308 (SD 108) v. 72 (SD 53) mg) and folic acid (412 (SD 89 μg) v. 187 (SD 107)) were recorded for UC patients compared with controls. Significant correlations were found for the consumption of different food groups and the chromosome index for chromosome 17. The results of our study suggest that the nutritional habits adopted by UC patients during clinical remission may affect key cellular components of the colonic tissue, inducing a high degree of aneuploidy and genetic instability, and probably affecting the development of colon cancer.

Epidemiologic studies have applied the glycaemic index (GI) and glycaemic load (GL) to assessments of usual dietary intake. Results have been inconsistent particularly for the association of GI or GL with diabetes incidence. We aimed to advance understanding of the GI and GL as applied to food frequency questionnaires (FFQ) by evaluating GI and GL in relation to plasma measures of glycaemia. Included were 1255 adults at a baseline examination (1994–6) and 813 who returned for the 5-year follow-up examination. Usual diet, at both examinations, was assessed by a validated FFQ. GI and GL were evaluated in relation to average fasting glucose (two measures at each examination) and 2h post-75g glucose load plasma glucose (baseline and follow-up), and glycated haemoglobin (A1c; follow-up only); using generalized linear models. Correlation coefficients (r) for GI and GL related to measures of glycaemia, adjusted for total energy intake, ranged from −0·004 to 0·04 (all NS) for both examinations. Adjustment for potential confounders, for fasting glucose in models for 2h glucose (to model incremental glucose) and for average fasting glucose in models for A1c (to account, in part, for overnight endogenous glucose production) also did not materially alter findings, nor did inclusion of data from both examinations together in linear mixed models. The present results call into question the utility of GI and GL to reflect glycaemic response to food adequately, when used in the context of usual diet. Further work is needed to quantify usual dietary exposures relative to glucose excursion and associated chronic glycaemia and other metabolic parameters.

Current anthropometric indices for health risk assessment are indirect measures of total or visceral body fat mass that do not consider the inverse relationship of lean body mass to metabolic risk as well as the non-linear relationship between central obesity and insulin resistance.We examined a new anthropometric index that reflects the relationship of waist circumference (WC) as a risk factor to fat-free mass (FFM) as a protective parameter of body composition. In apopulation of 335 adults (191 females and 144 males; mean age 53 (sd 13·9) years) with ahigh prevalence of obesity (27%) and metabolic syndrome (30%) we derived FFM:WC3 from the best fit of the relationship with metabolic risk factors (plasma triacylglycerol levels and insulin resistance by homeostasis model assessment index). Because FFM is known to be proportional to the cube of height, FFM was subsequently replaced by height3 yielding height3:WC3 as an easily applicable anthropometric index. Significant inverse relationships of height3:WC3 to metabolic risk factorswere observed for both sexes. They slightly exceeded those of conventional anthropometric indicessuch as BMI, WC or WC:hip ratio in women but not in men. The exponential character of the denominator WC3 implies that at a given FFM with gradually increasing WC the increasein metabolic risk is lower than proportional. Further studies are needed to evaluate height3:WC3 as an anthropometric index for health risk assessment.

I deficiency is the leading cause of preventable mental retardation. A number of surveys in Afghanistan show goitre prevalence rates more than 20% amongst children and women. Access to iodised salt remains low, with disparate coverage by region, despite the recent implementation of a national salt iodisation programme. The objectives were to identify whether the presence of goitre is a satisfactory marker of I deficiency and to examine the relationship between goitre and thyroid function. A case–control study was carried out in children and women of childbearing age, stratified on the presence of goitre. Adequate levels of urinary I were observed in 6·8% of all the subjects, and amongst the subjects without goitre, this figure was only 9%. The presence of goitre was significantly associated with severe urinary I deficiency; however, the difference between the cases and controls was not as great as expected. An association between the presence of goitre and elevated thyroid-stimulating hormone (TSH) levels was observed, but 14% of the children without palpable goitre also showed abnormal TSH levels.Given that the majority of subjects showed some degree of I deficiency and that children without goitre may have elevated TSH levels, the absence of goitre is an insufficient indicator to determine adequate I status. The risk of subsequent development of goitre, in the currently non-goitre population, is elevated. This suggests that short-term I supplementation should be considered independently of the presence of goitre or urinary I level, until the access to and consumption of iodised salt is generalised.

Previous studies have shown stimulatory effects of linoleic acid (LA, C18:2) on differentiation of rat muscle cells in culture (Allen et al.1985), but there appears to be little investigation of the effects of other fatty acids. The present study therefore compared the effects of different fatty acids on muscle cell differentiation in vitro. L6 myoblasts were cultured (Dulbecco's Modified Eagles Medium+10% fetal calf serum) in six-well plates until 80% confluent (day 0). Cells were then either harvested or the medium switched to differentiation medium (Dulbecco's Modified Eagles Medium+2% horse serum), supplemented with fatty acid or drug treatments. Cells were harvested on days 0–5 and assayed for creatine kinase (CK), protein and DNA contents, to give a measure of differentiation (CK/DNA). Initial studies indicated a stimulatory effect of the cis9trans11 (c9, t11) isomer of conjugated linoleic acid (CLA) relative to control. By contrast, the trans10, cis12 (t10, c12) isomer of CLA inhibited differentiation. Further experiments indicated that inhibition of differentiation by the t10, c12 CLA isomer was dose-dependent (up to 200μm) and may be via increased cell proliferation. LA and c9, t11 CLA stimulated differentiation at low concentrations (up to 50μm), but inhibited differentiation at high concentrations (200μm). In contrast, oleic acid stimulated differentiation at all concentrations, whereas the saturated fatty acid, palmitic acid, had no effect. The mechanism appeared not to involve either peroxisome proliferator-activated receptors α or γ. The data suggest that only unsaturated fatty acids have an effect and the presence or absence of a cis-9 double bond may be important.

The aim of the present study was: (1) to develop a new method for total energy expenditure (TEE) assessment, using accelerometry (ACC) and heart rate (HR) telemetry in combination; (2) to validate the new method against the criterion measure (DLW) and to compare with two of the most common methods, FLEX-HR and ACC alone. In the first part of the study VO2, HR and ACC counts were measured in twenty-seven subjects during walking and running on a treadmill. Considering the advantages and disadvantages of the HR and ACC methods an analysis model was developed, using ACC at intensities of low and medium levels and HR at higher intensities. During periods of inactivity, RMR is used. A formula for determining TEE from ACC, HR and RMR was developed: TEE =1·1×(EQHR×TTHR+EQACC1×TTACC1+EQACC2×TTACC2+RMR×TTRMR). In the validation part of the study a sub-sample of eight subjects wore an accelerometer, HR was logged and TEE was measured for 14d with the DLW method. Analysis of the Bland–Altman plots with 95% CI indicates that there are no significant differences in TEE estimated with HR–ACC and ACC alone compared with TEE measured with DLW. It is concluded that the HR–ACC combination as well as ACC alone has potential as a method for assessment of TEE during free-living activities as compared with DLW

The principal phyto-oestrogens (PO) in food are isoflavones, lignans, coumestans and prenylated flavonoids, with isoflavones and lignans being the most commonly found in UK diets. Until recently obtaining accurate data on the PO content of foods was hampered by lack of suitable analytical methods and validation techniques. Furthermore, although PO data exist for some foods, these foods may not be available in the UK. The aim of the present study was to construct a new, comprehensive isoflavone (total genistein + daidzein) database. Using data, mainly from recent GC–MS analysis, for approximately 300 foods available in the UK, and extensive recipe calculations, a new database was constructed containing approximately 6000 foods allocated an isoflavone value. By analysing 7d weighed food diaries, the database was subsequently used to estimate isoflavone intake in two groups of healthy volunteers, omnivores (n 9) and vegetarians (n 10). Mean isoflavone intake in the vegetarian and omnivorous group was 7·4 (sem 3·05) and 1·2 (sem 0·43) mg/d, respectively. Mean intake for the total group was 4·5 (sem 1·89) mg/d. Main food sources of isoflavones for the vegetarian group were soya milk (plain), meat-substitute foods containing textured vegetable protein and soya protein isolate, soya mince, wholemeal bread and rolls, white bread and rolls, croissants and pitta breads, beans, raisins and soya sauce. Main food sources of isoflavones for the omnivorous group were soya yogurts, wholemeal bread and rolls, white bread and rolls, garlic bread, nan bread and brown bread, sultanas and scones.

Breast cancer is the most common cancer in women and a significant cause of death. Mutations of the oncosuppressor genes BRCA1 and BRCA2 are associated with a hereditary risk of breast cancer, and dysregulation of their expression has been observed in sporadic cases. Soya isoflavones have been shown to inhibit breast cancer in studies in vitro, but associations between the consumption of isoflavone-containing foods and breast cancer risk have varied in epidemiological studies. Soya is a unique source of the phytoestrogens daidzein (4′,7-dihydroxyisoflavone) and genistein (4′,5,7-trihydroxyisoflavone), two molecules that are able to inhibit the proliferation of human breast cancer cells in vitro. The aim of the present study was to determine the effects of genistein (5μg/ml) and daidzein (20μg/ml) on transcription in three human breast cell lines (one dystrophic, MCF10a, and two malignant, MCF-7 and MDA-MB-231) after 72h treatment. The different genes involved in the BRCA1 and BRCA2 pathways (GADD45A, BARD1, JUN, BAX, RB1, ERα, ERβ, BAP1, TNFα, p53, p21Waf1/Cip1, p300, RAD51, pS2, Ki-67) were quantified by real-time quantitative RT-PCR, using the TaqMan method and an ABI Prism 7700 Sequence Detector (Applied Biosystems). We observed that, in response to treatment, many of these genes were overexpressed in the breast cancer cell lines (MCF-7 and MDA-MB-231) but not in the dystrophic cell line (MCF10a).

Maillard products arise from condensation reactions between amino acids or proteins with reducing sugars during food processing. As ubiquitous components of human food, these early or advanced glycation products may be subject to intestinal absorption. The present study was performed to investigate the intestinal uptake of Maillard products and to determine whether they are substrates for peptide and amino acid transporters expressed at the apical membrane of Caco-2 cells. At a concentration of 10mm, Nɛ-(carboxymethyl)-L-lysine, Nα-hippuryl-Nɛ-(1-deoxy-d-fructosyl)-l-lysine,Nα-hippuryl-Nɛ-(carboxymethyl)-L-lysine and Nɛ-(1-deoxy-d-fructosyl)-l-lysine inhibited the [14C]glycylsarcosine uptake mediated by the H+–peptide co-transporter PEPT1 by 13 to 45%.For Nɛ-(1-deoxy-d-fructosyl)-l-lysine, an inhibitory constant of 8·7mm was determined, reflecting a low affinity to PEPT1 in comparison with natural dipeptides. Uptake of l-[3H]lysine was weakly affected by Nɛ-(carboxymethyl)-L-lysine, Nα-hippuryl-l-lysine and Nα-hippuryl-Nɛ-(carboxymethyl)-L-lysine but strongly inhibited by Nɛ-(1-deoxy-d-fructosyl)-l-lysine (81%). None of the Maillard products was able to inhibit the uptake of l-[3H]leucine by more than 15%. We also studied the transepithelial flux of Maillard productsacross Caco-2 cell monolayers cultured on permeable filters. The flux rates of Maillard products ranged from 0·01 to 0·3%/cm2 per h and were shown to be muchlower than those of carrier substrates such as glycylsarcosine, l-proline and the space marker 14C]mannitol. We conclude that the Maillard products investigated in the present study are neither transported by PEPT1 nor by carriers for neutral amino acids. The low transepithelial flux measured for these compounds most probably occurs by simple diffusion.

The ingestion of a glucose-containing drink has been shown to improve cognitive performance, particularly memory functioning. However, it remains unclear as to the extent to which task domain and task difficulty moderate the glucose enhancement effect. The aim of this research was to determine whether boosts in performance are restricted to particular classes of memory (episodic v. semantic) or to tasks of considerable cognitive load. A repeated measures (25g glucose v saccharin), counterbalanced, double-blind design was used with younger and older adults. Participants performed a battery of episodic (e.g. paired associate learning) and semantic memory (e.g. category verification) tasks under low and high cognitive load. Electrophysiological measures (heart rate and galvanic skin response) of arousal and mental effort were also gathered. The results indicated that whilst glucose appeared to aid episodic remembering, cognitive load did not exaggerate the facilitative effect. For semantic memory, there was little evidence to suggest that glucose can boost semantic memory retrieval even when the load was manipulated. One exception was that glucose facilitated performance during the difficult category fluency task. Regardless, the present findings are consistent with the domain-specific account in which glucose acts primarily on the hippocampal region, which is known to support episodic memory. The possible contribution of the hippocampus in semantic memory processing is also discussed.

The aim of the present study was to determine the accuracy of reported energy intake according to a food-frequency questionnaire (FFQ) and dietary records (DR) in obese subjects with metabolic syndrome risk factors. Subjects were twenty-three men and twenty-seven women with mean BMI of 35·7 (range 30·5–43·8) kg/m2 who participated in a dietary interview based on a FFQ and completed weighed DR. Total energy expenditure was measured with the doubly labelled water method. Total energy expenditure, measured RMR and physical activity level did not differ between under-reporters (50% of the sample) and non-under-reporters. Under-reporters had lower median intake of sweets, desserts and snacks than non-under-reporters (100 V.. 161g/d (P=0·0008) and 61 V.. 128g/d (P.=0·0002) according to the FFQ and DR, respectively). The DR also showed lower energy density (6·7 (sd 1·3) V 7·9 (sd 1·6) kJ/g; P=0·0064), lower intake of sugary drinks (0 V. 167g/d; P=0·0063) and higher scores for dietary restraint (9·0 (sd 5·0) V. 6·1 (sd 3·5); P=0·0285) in under-reporters. Energy density was associated with accuracy according to the FFQ (Spearman's rank correlation coefficient (RS) 0·406; P=0·0034) and the DR (RS 0·537; P<0·0001). In multivariate analysis, consumption of bread and sweets, desserts and snacks measured by the FFQ was positively associated with accuracy (I2adjusted 0·46 (95% CI 0·32, 0·70)). According to the DR, consumption of sweets, desserts and snacks was also associated with accuracy, as was dietary restraint (inversely) (I2adjusted 0·67 (95% CI 0·54, 0·83)). In obese subjects with metabolic risk factors, intake of sweets, desserts and snacks, bread and dietary restraint were determinants of reporting accuracy.

The composition and activities of the faecal microbiota in twelve healthy subjects analysed in a single open study were monitored before (1-week baseline step), during (10d supplementation step) and after (10d follow-up step) the ingestion of a fermented milk containing Lactobacillus casei DN-114001. Fluorescent in situ hybridisation with group-specific DNA probes, real-time PCR using L. paracasei group-specific primers and temporal temperature gradient gel electrophoresis (TTGE) using group-specific primers were carried out, together with bacterial enzyme activity and metabolite analyses to monitor the structure and activities of the faecal microbiota. L. casei DNA was detected in the faeces of all of the subjects by TTGE after 10d supplementation. Its quantification by real-time PCR showed a 1000-fold increase during the test step compared with initial levels. No major modification in either the dominant members of the faecal microbiota or their activities was observed during the trial. In conclusion, the short-term consumption of a milk product containing L. casei DN-114001 was accompanied by a high, transient increase in the quantity of this strain in the faeces of all of the subjects without markedly affecting biochemical or bacteriological factors.