European Journal of Nutrition (v.52, #2)

Vitamin D metabolism, functions and needs: from science to health claims by S. Battault; S. J. Whiting; S. L. Peltier; S. Sadrin; G. Gerber; J. M. Maixent (429-441).
Vitamin D is a nutrient long considered as essential for skeletal health but is now attracting interest from medical and nutritional communities as knowledge emerges of its biological function and its association with decreased risk of many chronic diseases.A question emerges: How much more vitamin D do we need for these new functions of vitamin D? This review discusses vitamin D physiology and hypovitaminosis D and presents two vitamin D dietary policies: that according to regulatory authorities and that of nutrition scientists. Scientific evidence suggests that 25(OH)D serum levels should be over 75 nmol/L; otherwise, there is no beneficial effect of vitamin D on long-latency diseases. Current regulatory authority recommendations are insufficient to reach this level of adequacy. Observational and some prospective data show that vitamin D has a role in the prevention of cancer as well as immunity, diabetes and cardiovascular and muscle disorders, which supports the actions of 1α,25(OH)2D at cellular and molecular levels. The recent assessments done by the European Food Safety Authority should lead to new health claims.Vitamin D, through food fortification and supplementation, is a promising new health strategy and thus provides opportunities for food industry and nutrition researchers to work together towards determining how to achieve this potential health benefit.
Keywords: Vitamin D; 25(OH)D; Supplementation; DRI; Health claim

Oil palm phenolics attenuate changes caused by an atherogenic diet in mice by Soon-Sen Leow; Shamala Devi Sekaran; Kalyana Sundram; YewAi Tan; Ravigadevi Sambanthamurthi (443-456).
Water-soluble phenolics from the oil palm possess significant biological properties.In this study, we aimed to discover the role of oil palm phenolics (OPP) in influencing the gene expression changes caused by an atherogenic diet in mice.We fed mice with either a low-fat normal diet (14.6 % kcal/kcal fat) with distilled water, or a high-fat atherogenic diet (40.5 % kcal/kcal fat) containing cholesterol. The latter group was given either distilled water or OPP. We harvested major organs such as livers, spleens and hearts for microarray gene expression profiling analysis. We determined how OPP changed the gene expression profiles caused by the atherogenic diet. In addition to gene expression studies, we carried out physiological observations, blood hematology as well as clinical biochemistry, cytokine profiling and antioxidant assays on their blood sera.Using Illumina microarrays, we found that the atherogenic diet caused oxidative stress, inflammation and increased turnover of metabolites and cells in the liver, spleen and heart. In contrast, OPP showed signs of attenuating these effects. The extract increased unfolded protein response in the liver, attenuated antigen presentation and processing in the spleen and up-regulated antioxidant genes in the heart. Real-time quantitative reverse transcription-polymerase chain reaction validated the microarray gene expression fold changes observed. Serum cytokine profiling showed that OPP attenuated inflammation by modulating the Th1/Th2 axis toward the latter. OPP also increased serum antioxidant activity to normal levels.This study suggests that OPP may possibly attenuate atherosclerosis and other forms of cardiovascular disease.
Keywords: Oil palm phenolics; Antioxidants; Atherosclerosis; Cardiovascular disease; Microarray; Gene expression

Active hexose-correlated compound and Bifidobacterium longum BB536 exert symbiotic effects in experimental colitis by Borja Ocón; Andrea Anzola; Mercedes Ortega-González; Antonio Zarzuelo; María D. Suárez; Fermín Sánchez de Medina; Olga Martínez-Augustin (457-466).
Active hexose-correlated compound (AHCC) is a commercial extract obtained from Basidiomycetes under controlled conditions, yielding a 74 % content in oligosaccharides, especially α-glucans. AHCC has a number of therapeutic effects, including intestinal anti-inflammatory activity. Bifidobacterium longum BB536 is a probiotic with potential health-promoting effect at the gut level. The purpose of the present study was to evaluate the possibility of synergism between AHCC, which is believed to act as a prebiotic, and B. longum BB536.We used the trinitrobenzene sulfonic acid model (TNBS) of colitis in rats. AHCC (100 or 500 mg kg−1) and B. longum BB536 (5 × 106 CFU rat−1 day−1) were administered together or separately for 7 days prior to colitis induction and then for another 7 days and compared with control (noncolitic) and TNBS rats.The results show that both treatments had intestinal anti-inflammatory activity separately, which was enhanced when used in combination, as shown by changes in body weight gain, colonic weight to length ratio, myeloperoxydase activity and iNOS expression. Interestingly, the association of AHCC 100 mg kg−1 + B. longum BB536 showed the highest anti-inflammatory activity.Our data provide a preclinical experimental basis for the synergistic effect of AHCC and B. longum BB536 on inflammatory bowel disease.
Keywords: Active hexose-correlated compound; Bifidobacterium longum BB536; Symbiotic; Colitis

Suboptimal iron status and associated dietary patterns and practices in premenopausal women living in Auckland, New Zealand by Kathryn L. Beck; Rozanne Kruger; Cathryn A. Conlon; Anne-Louise M. Heath; Christophe Matthys; Jane Coad; Welma Stonehouse (467-476).
To investigate associations between dietary patterns and suboptimal iron status in premenopausal women living in Auckland, New Zealand.Premenopausal women (n = 375; 18–44 years) were included in this cross-sectional analysis. Suboptimal iron status was defined as serum ferritin <20 μg/L. Participants completed a 144-item iron food frequency questionnaire (FeFFQ) and a questionnaire on dietary practices to assess dietary intake over the past month. Factor analysis was used to determine dietary patterns from the FeFFQ. Logistic regression was used to determine associations between these dietary patterns and iron status.Seven dietary patterns were identified: refined carbohydrate and fat; Asian; healthy snacks; meat and vegetable; high tea and coffee; bread and crackers; and milk and yoghurt. Logistic regression suggested that following a “meat and vegetable” dietary pattern reduced the risk of suboptimal iron status by 41 % (95 % CI: 18, 58 %; P = 0.002) and following a “milk and yoghurt” pattern increased the risk of suboptimal iron status by 50 % (95 % CI: 15, 96 %; P = 0.003).These results suggest that dietary patterns characterized by either a low intake of meat and vegetables or a high intake of milk and yoghurt are associated with an increased risk of suboptimal iron status. Dietary pattern analysis is a novel and potentially powerful tool for investigating the relationship between diet and iron status.
Keywords: Iron; Dietary patterns; Factor analysis; Women

The aim of the study was to examine the acute effects of pre-exercise ingestion of protein, carbohydrate, and a non-caloric placebo on serum concentrations of insulin and cortisol, and the intramuscular gene expression of myostatin- and ubiquitin proteasome pathway (UPP)-related genes following a bout of resistance exercise.Ten untrained college-aged men participated in three resistance exercise sessions (3 × 10 at 80 % 1RM for bilateral hack squat, leg press, and leg extension) in a cross-over fashion, which were randomly preceded by protein, carbohydrate, or placebo ingestion 30 min prior to training. Pre-supplement/pre-exercise, 2 h and 6 h post-exercise muscle biopsies were obtained during each session and analyzed for mRNA fold changes in myostatin (MSTN), activin IIB, follistatin-like 3 (FSTL3), SMAD specific E3 ubiquitin protein ligase 1 (SMURF1), forkhead box O3, F-box protein 32 (FBXO32), and Muscle RING-finger protein-1, with beta-actin serving as the housekeeping gene. Gene expression of all genes was analyzed using real-time PCR.Acute feeding appeared to have no significant effect on myostatin or UPP biomarkers. However, resistance exercise resulted in a significant downregulation of MSTN and FBXO32 mRNA expression and a significant upregulation in FSTL3 and SMURF1 mRNA expression (p < 0.05).An acute bout of resistance exercise results in acute post-exercise alterations in intramuscular mRNA expression of myostatin and UPP markers suggestive of skeletal muscle growth. However, carbohydrate and protein feeding surrounding resistance exercise appear to have little influence on the acute expression of these markers.
Keywords: Hypertrophy; Strength; Insulin; Cortisol; Protein

Leg fat might be more protective than arm fat in relation to lipid profile by M. Sánchez-López; F. B. Ortega; P. Moya-Martínez; S. López-Martínez; I. Ortiz-Galeano; M. A. Gómez-Marcos; M. Sjöström; V. Martínez-Vizcaíno (489-495).
The objective of this study was to determine the independent relationships of trunk fat, leg fat and arm fat to cardiovascular (CVD) risk factors, after controlling for relevant confounders such as fat mass index, cardiorespiratory fitness and objectively measured physical activity.This is a cross-sectional study involving 683 university students, aged 18–30 years. Total and regional body fat distribution was measured using dual-energy X-ray absorptiometry. The associations of trunk, leg and arm fat with CVD risk factors (triglycerides-TG-, high-density lipoprotein cholesterol-HDL-c-, TG/HDL-c ratio, HOMAIR, mean arterial pressure, C-reactive protein) were examined using regression linear models, controlling for age, sex, fat mass index [total body fat(kg)/height(m2)], maximal oxygen consumption and physical activity by accelerometer.After controlling for fat mass index, and other confounders, higher levels of trunk fat were found to be associated with a poorer lipid profile, while higher levels of leg fat were found to be associated with a better lipid profile. We did not find any association between arm fat and lipid profile after controlling for total fatness and other confounders. Neither trunk, leg or arm fat was found to be related to insulin resistance, blood pressure or inflammation markers.Our data suggest that the region where fat is accumulated might have a differential effect on lipid profile: trunk fat has an adverse effect, leg fat has a protective effect, and arm fat has no effect. The differences observed between upper- and lower-body peripheral fat depots should be further explored.
Keywords: Cardiovascular risk factors; Fat distribution; Cardiorespiratory fitness and physical activity

Factors affecting the distribution of folate forms in the serum of elderly German adults by Susanne H. Kirsch; Wolfgang Herrmann; Rudolf Eckert; Jürgen Geisel; Rima Obeid (497-504).
We investigated the roles of age, vitamin B12 markers, and the 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism as determinants of folate forms in serum.We measured the serum concentrations of (6S)-5-CH3–H4folate, (6S)-H4folate, (6S)-5-HCO–H4folate, (6R)-5,10-CH+–H4folate, and folic acid in 146 non-supplemented older participants (median age 74 years). The concentrations of total vitamin B12, holotranscobalamin (holoTC), methylmalonic acid (MMA), and total homocysteine (tHcy) were also measured.Elevated metabolites (MMA > 271 nmol/L and tHcy > 12.0 μmol/L) were found in 24.0 and 63.0 % of the participants, respectively. We found a significant age-dependent decrease (participants with a median age of 87 years compared with participants with a median age of 60 years) in the sum of serum folate levels, the (6S)-5-CH3–H4folate concentration, and the (6S)-5-CH3–H4folate proportion. In addition, participants with elevated metabolite levels were older, had lower concentrations of the sum of folates and (6S)-5-CH3–H4folate, and had higher concentrations of (6S)-5-CHO–H4folate and creatinine but had a comparable holoTC/total vitamin B12 ratio. No association was found between the MTHFR C677T genotype and serum folate forms.Low serum (6S)-5-CH3–H4folate concentrations and the proportion of (6S)-5-CH3–H4folate (percentage of the sum of folate forms) are related to older age and elevated MMA and tHcy levels.
Keywords: Folate; Folic acid; Age; Elderly; Vitamin B12

Combined arginine and glutamine decrease release of de novo synthesized leukotrienes and expression of proinflammatory cytokines in activated human intestinal mast cells by Sandra Lechowski; Katharina Feilhauer; Ludger Staib; Moïse Coëffier; Stephan C. Bischoff; Axel Lorentz (505-512).
PurposeGlutamine and arginine modulate inflammatory responses of epithelial cells and monocytes. Here, we studied the response of human mast cells to pharmacological doses of arginine and glutamine.Mast cells isolated from intestinal tissue were incubated with physiological doses of arginine (0.1 mmol/L) and glutamine (0.6 mmol/L) or with pharmacological doses of arginine (2 mmol/L) and glutamine (10 mmol/L) for 18 h. Following stimulation by IgE receptor crosslinking mast cell mediators were measured by enzymatic assay, ELISA, multiplex bead immunoassay, or real-time RT-PCR, and activation of intracellular signaling molecules was determined using proteome profiler array or immunoblotting.We found that the combined challenge of mast cells with pharmacological doses of arginine and glutamine caused a decrease in induced release of de novo synthesized leukotriene C4 but not of pre-stored β-hexosaminidase. Moreover, we found reduced expression of chemokines monocyte chemoattractant protein-1 (CCL2), macrophage inflammatory protein-1β (CCL4), IL-8 (CXCL8), and TNF in response to high doses of both amino acids. The anti-inflammatory effects of arginine and glutamine were associated with decreased activation levels of signaling molecules known to be involved in mast cell cytokine expression such as MAPK family members extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38, and the protein kinase B (Akt).Arginine and glutamine attenuate IgE-dependent human mast cell activation by decreasing lipid mediator release and expression of proinflammatory cytokines.
Keywords: Arginine; Glutamine; Intestinal mucosa; Mast cell; Cytokine expression

Vitamin D status is associated with sociodemographic factors, lifestyle and metabolic health by Tuija Jääskeläinen; Paul Knekt; Jukka Marniemi; Laura Sares-Jäske; Satu Männistö; Markku Heliövaara; Ritva Järvinen (513-525).
Low serum 25(OH)D concentration has been shown to predict the occurrence of several chronic diseases. It is, however, still unclear whether the associations are causal or due to confounding. The aim of this study was to investigate the associations between serum 25(OH)D concentration and sociodemographic, lifestyle and metabolic health-related factors.The study population comprised 5,714 men and women, aged 30–79 years, from the Health 2000 Survey representing the Finnish population. Serum 25(OH)D concentration was determined by radioimmunoassay from serum samples frozen at −70 °C. Sociodemographic, lifestyle and metabolic factors were determined by questionnaires, interviews and measurements. Linear regression was used to assess the associations between serum 25(OH)D and the factors studied.The mean serum 25(OH)D concentration was 45.3 nmol/l and it varied between categories of sociodemographic, lifestyle and metabolic health variables. Older age, being married or cohabiting and higher education were related to higher serum 25(OH)D concentration. Those with the healthiest lifestyle estimated by a lifestyle index based on body mass index, physical activity, smoking, alcohol consumption and diet had 15.8 nmol/l higher serum 25(OH)D concentration compared to those with the unhealthiest lifestyle. Of the indicators of metabolic health, only waist circumference and HDL cholesterol were significantly associated with 25(OH)D after adjustment for sociodemographic, lifestyle and other metabolic health factors.This study suggests that serum 25(OH)D concentration is associated with a multitude of sociodemographic, lifestyle and metabolic health factors. Thus, it is possible that such factors confound associations observed between serum 25(OH)D concentration and chronic diseases.
Keywords: Vitamin D status; Serum 25(OH)D; Sociodemographic factors; Lifestyle; Metabolic health; Cross-sectional study

A moderate weight reduction through dietary intervention decreases hepatic fat content in patients with non-alcoholic fatty liver disease (NAFLD): a pilot study by Valentina Volynets; Jürgen Machann; Markus A. Küper; Ina B. Maier; Astrid Spruss; Alfred Königsrainer; Stephan C. Bischoff; Ina Bergheim (527-535).
As a diet rich in fructose and an impaired intestinal barrier function have been proposed to be risk factors for the development of non-alcoholic fatty liver disease (NAFLD), the aim of the present pilot study was to determine whether a dietary intervention focusing on a reduction of fructose intake (−50 % in comparison with baseline) has a beneficial effect on liver status.A total of 15 patients with NAFLD were enrolled in the study of which 10 finished the study. Fructose and total nutrient intake were assessed using a diet history. At baseline and after 6 months liver status and markers of intestinal barrier function as well as plasminogen activator inhibitor (PAI-) 1 concentration were determined in plasma.Hepatic lipid content and transaminases in plasma as well as body mass index and some parameters of glucose metabolism (e.g., fasting plasma insulin) were significantly lower at the end of the intervention when compared to baseline. Whereas the dietary intervention had no effect on the prevalence of bacterial overgrowth, orocecal transit time and the intestinal permeability or blood ethanol levels endotoxin and PAI-1 concentration in plasma were significantly lower at the end of 6 months intervention period than at baseline.Taken together, our results indicate that a dietary intervention focusing only on one dietary parameter like fructose may help to decrease intrahepatic fat content of NAFLD patients.
Keywords: NAFLD; Endotoxin; Fructose; PAI-1; Metabolic syndrome

Increased hepatic de novo lipogenesis and mitochondrial efficiency in a model of obesity induced by diets rich in fructose by Raffaella Crescenzo; Francesca Bianco; Italia Falcone; Paola Coppola; Giovanna Liverini; Susanna Iossa (537-545).
To assess hepatic de novo lipogenesis and mitochondrial energetics as well as whole-body energy homeostasis in sedentary rats fed a fructose-rich diet.Male rats of 90 days of age were fed a high-fructose or control diet for 8 weeks. Body composition, energy balance, oxygen consumption, carbon dioxide production, non-protein respiratory quotient, de novo lipogenesis and insulin resistance were measured. Determination of specific activity of hepatic enzymes of de novo lipogenesis, mitochondrial mass, oxidative capacity and degree of coupling, together with parameters of oxidative stress and antioxidant defence, was also carried out.Body energy and lipid content as well as plasma insulin and non-esterified fatty acids were significantly higher in fructose-fed than in control rats. Significantly higher rates of net de novo lipogenesis and activities of hepatic lipogenic enzymes fatty acid synthase and stearoyl CoA desaturase-1 were found in fructose-fed rats compared to controls. Mitochondrial protein mass and degree of coupling were significantly higher in fructose-fed rats compared to controls. Hepatic mitochondria showed oxidative damage, both in the lipid and in the protein component, together with decreased activity of antioxidant defence.Liver mitochondrial compartment is highly affected by fructose feeding. The increased mitochondrial efficiency allows liver cells to burn less substrates to produce ATP for de novo lipogenesis and gluconeogenesis. In addition, increased lipogenesis gives rise to whole body and ectopic lipid deposition, and higher mitochondrial coupling causes mitochondrial oxidative stress.
Keywords: Mitochondrial uncoupling; Insulin resistance; Body lipids; De novo lipogenesis

LDL-cholesterol-lowering effect of a dietary supplement with plant extracts in subjects with moderate hypercholesterolemia by Nicolas Ogier; Marie-Josèphe Amiot; Stéphane Georgé; Matthieu Maillot; Cécilia Mallmann; Marie Maraninchi; Sophie Morange; Jean-François Lescuyer; Sébastien L. Peltier; Nicolas Cardinault (547-557).
Red yeast rice (RYR), sugar cane-derived policosanols (SCdP) and artichoke leaf extracts (ALEs) are currently incorporated alone or in combination into dietary supplements for their potential low-density-lipoprotein cholesterol (LDL-cholesterol)-lowering effects. Yet, there is no information supporting the efficacy of this association on the reduction in LDL-cholesterol. The main objective of this study was to investigate the effects of a new dietary supplement (DS) with RYR, SCdP and ALEs on LDL-cholesterol.In a double-blind, randomized, parallel controlled study, 39 subjects from 21 to 55 years with moderate hypercholesterolemia without drug treatment were assigned to 2 groups and then consumed either a DS containing RYR, SCdP and ALEs or a placebo over a 16-week period. Plasma concentrations of lipids [LDL-cholesterol, total cholesterol (TC), high-density-lipoprotein cholesterol (HDL-cholesterol), triacylglycerols (TG)] and plasma levels of vitamins C and E, total polyphenols and malondialdehyde were determined at baseline and after 4, 8, 12 and 16 weeks.LDL-cholesterol and TC were reduced by, respectively, 21.4 % (95 % CI, −13.3 to −24.9 %, p < 0.001) and 14.1 % (95 % CI, −10.1 to −18.0 %, p < 0.001) at week 16 in the DS group compared with baseline. Similar results were obtained at weeks 4, 8 and 12. TG decreased by 12.2 % after 16 weeks in the DS group (95 % CI: −24.4 to −0.1 %, p < 0.05). For the vitamin E/TC ratio, a difference was observed between groups at week 16 (p < 0.05). Other parameters were not modified.Daily consumption of this new DS decreased LDL-cholesterol and TC and is therefore an interesting, convenient aid in managing mild to moderate hypercholesterolemia.
Keywords: Dietary supplement; Low-density-lipoprotein cholesterol; Lipid-lowering plant extracts; Red yeast rice; Policosanols; Hypercholesterolemia

Fruits and nuts may prevent or reverse common human health conditions such as obesity, diabetes and hypertension; together, these conditions are referred to as metabolic syndrome, an increasing problem. This study has investigated the responses to ellagic acid, present in many fruits and nuts, in a diet-induced rat model of metabolic syndrome.Eight- to nine-week-old male Wistar rats were divided into four groups for 16-week feeding with cornstarch diet (C), cornstarch diet supplemented with ellagic acid (CE), high-carbohydrate, high-fat diet (H) and high-carbohydrate, high-fat diet supplemented with ellagic acid (HE). CE and HE rats were given 0.8 g/kg ellagic acid in food from week 8 to 16 only. At the end of 16 weeks, cardiovascular, hepatic and metabolic parameters along with protein levels of Nrf2, NF-κB and CPT1 in the heart and the liver were characterised.High-carbohydrate, high-fat diet-fed rats developed cardiovascular remodelling, impaired ventricular function, impaired glucose tolerance, non-alcoholic fatty liver disease with increased protein levels of NF-κB and decreased protein levels of Nrf2 and CPT1 in the heart and the liver. Ellagic acid attenuated these diet-induced symptoms of metabolic syndrome with normalisation of protein levels of Nrf2, NF-κB and CPT1.Ellagic acid derived from nuts and fruits such as raspberries and pomegranates may provide a useful dietary supplement to decrease the characteristic changes in metabolism and in cardiac and hepatic structure and function induced by a high-carbohydrate, high-fat diet by suppressing oxidative stress and inflammation.
Keywords: Cardiovascular remodelling; Ellagic acid; Metabolic syndrome; Non-alcoholic fatty liver disease; Obesity

Vitamin C-related nutrient–nutrient and nutrient–gene interactions that modify folate status by Mark Lucock; Zoë Yates; Lyndell Boyd; Charlotte Naylor; Jeong-Hwa Choi; Xiaowei Ng; Virginia Skinner; Ron Wai; Jeremy Kho; Sa Tang; Paul Roach; Martin Veysey (569-582).
Folate-related nutrient–nutrient and nutrient–gene interactions modify disease risk; we therefore examined synergistic relationships between dietary folic acid, vitamin C and variant folate genes with respect to red cell folate status.Two hundred and twelve subjects were examined using chemiluminescent immunoassay, PCR and food frequency questionnaire to determine red cell and serum folate, 14 folate gene polymorphisms, dietary folate (natural and synthetic) and vitamin C.When examined independently, synthetic PteGlu correlates best with red cell folate at higher levels of intake (p = 0.0102), while natural 5CH3-H4-PteGlun correlates best with red cell folate at lower levels of intake (p = 0.0035). However, dietary vitamin C and 5CH3-H4-PteGlun interact synergistically to correlate with red cell folate at higher levels of intake (p = 0.0005). No interaction between dietary vitamin C and PteGlu was observed. This ‘natural’ nutrient–nutrient interaction may provide an alternative to synthetic PteGlu supplementation that is now linked to adverse phenomena/health outcomes. On its own, vitamin C also correlates with red cell folate (p = 0.0150) and is strongly influenced by genetic variation in TS, MTHFR and MSR, genes critical for DNA and methionine biosynthesis that underpin erythropoiesis. Similarly, dietary vitamin C and 5CH3-H4-PteGlun act synergistically to modify red cell folate status according to variation in folate genes: of note, heterozygosity for 2R3R-TS (p = 0.0181), SHMT (p = 0.0046) and all three MTHFR SNPs (p = 0.0023, 0.0015 and 0.0239 for G1793A, C677T and A1298C variants, respectively) promote a significant association with red cell folate. Again, all these genes are critical for nucleic acid biosynthesis. Folate variants with the strongest independent effect on folate status were C677T-MTHFR (p = 0.0004) and G1793A-MTHFR (p = 0.0173).5CH3-H4-PteGlun assimilation and variant folate gene expression products may be critically dependent on dietary vitamin C.
Keywords: Antioxidants; Ascorbic acid; Folic acid; Polymorphism; Vitamin; Bioavailability

Vitamin A deficiency (VAD) and anemia are major challenges among children and expecting and lactating mothers in developing countries. Intervention with locally available dark green leafy vegetables (DGLV) is more sustainable to eradicate VAD, being cost-effective and readily adaptable to local communities. DGLV contain high levels of iron and β-carotene (BC) and therefore useful in fighting VAD and anemia. Since DGLVs are season-dependent sun-drying enables their availability during low seasons. However, their contribution to the bioavailability of BC and the improvement of hemoglobin are not well understood. The study therefore investigated the effect of consuming cooked recipe consisting of sun-dried amaranth and cowpea leaves on the levels of BC, retinol, and hemoglobin in preschool children from Machakos District, a semiarid region in Kenya.Vegetables were purchased from local vegetable market, with some sun-dried in an open shade. Levels of BC and retinol in serum and BC in fresh and processed vegetables were determined by a HPLC method and hemoglobin using a portable Hemocue Analyzer.All-trans-BC levels in uncooked fresh cowpea and amaranth leaves were 806.0 μg/g and 599.0 μg/g dry matter, respectively, while the dehydration and cooking processes retained the β-carotene levels at over 60 %. Consumption of the dehydrated vegetables significantly improved both serum BC and retinol levels (p < 0.05), while the baseline hemoglobin levels improved by 4.6 %.The study has shown that intervention with locally available sun-dried vegetables improves the bioavailability of BC, retinol, and hemoglobin levels among preschool children.
Keywords: β-carotene; Retinol; Hemoglobin; Vitamin A deficiency; Sun-dried vegetables

Antiplatelet effect of new lipophilic hydroxytyrosol alkyl ether derivatives in human blood by J. J. Reyes; J. P. De La Cruz; J. Muñoz-Marin; A. Guerrero; J. A. Lopez-Villodres; A. Madrona; J. L. Espartero; J. A. Gonzalez-Correa (591-599).
To investigate the in vitro antiplatelet and anti-inflammatory effects of five alkyl hydroxytyrosol (HT) ether derivatives in human whole blood and compare these effects with those of HT.Blood samples from healthy volunteers were incubated with HT and HT alkyl ether derivatives (ethyl, butyl, hexyl, octyl and dodecyl). Maximum intensity of platelet aggregation was induced with collagen, arachidonic acid or ADP. Calcium-induced thromboxane B2 and nitric oxide production, LPS-induced prostaglandin E2 and nitric oxide production and LPS-induced interleukin 1β production were measured.All compounds inhibited platelet aggregation, thromboxane B2 and inflammatory mediators in a concentration-dependent manner. The concentrations of each compound that inhibited the corresponding variable by 50 % compared to control samples (IC50) were in the range of 10−7–10−6 M for HT hexyl ether; for the other compounds, these values were in the range of 10−5 M. The IC50 for thromboxane B2 production was in the range of 10−4 M. The effects of HT alkyl ether derivatives were greater than those of HT. These compounds increased nitric oxide production. There was no direct relationship between the effects of these compounds and alkyl chain length. Maximum effects were observed in the C4–C6 range.Alkyl ether derivatives of HT exert antiplatelet and anti-inflammatory effects that are greater than those of HT.
Keywords: Hydroxytyrosol alkyl ether derivatives; Platelet aggregation; Prostanoids; Nitric oxide; Inflammatory mediators

Chlorella pyrenoidosa ameliorated L-NAME-induced hypertension and cardiorenal remodeling in rats by Su-Ching Yang; Hsin-Yi Yang; Yi-Ching Yang; Hsiang-Chi Peng; Pei-Yin Ho (601-608).
Hypertension is one of the main factors causing cardiovascular diseases. The aim of the study is to investigate the effects of Chlorella pyrenoidosa on blood pressure and cardiorenal remodeling in rats with N ω-nitro-l-arginine methyl ester hydrochloride (L-NAME)-induced endothelial dysfunction.Rats were fed a diet containing L-NAME (40 mg/kg) with or without chlorella (4 or 8 %) for 5 weeks. We found that chlorella retarded the development of hypertension and cardiorenal remodeling during the 5-week experimental period.Although there was no difference in NO x levels or plasma arginine concentrations, plasma and tissues ACE activities were significantly lower in the chlorella groups than in the L-NAME group. Moreover, tissue tumor necrosis factor-α concentrations and renal CYP4A expression were also lower in the chlorella group.These results suggest that chlorella might ameliorate the elevation of blood pressure and show cardiorenal-protective effects in nitric oxide-deficient rats, and one possible mechanism might be mediated by its ACE inhibitory activity.
Keywords: Chlorella; Hypertension; Nitric oxide; Angiotensin-converting enzyme

Dairy intake in relation to cardiovascular disease mortality and all-cause mortality: the Hoorn Study by Marieke A. van Aerde; Sabita S. Soedamah-Muthu; Johanna M. Geleijnse; Marieke B. Snijder; Giel Nijpels; Coen D. A. Stehouwer; Jacqueline M. Dekker (609-616).
Existing data from prospective cohort studies on dairy consumption and cardiovascular diseases are inconsistent. Even though the association between total dairy and cardiovascular diseases has been studied before, little is known about the effect of different types of dairy products on cardiovascular diseases (CVD). The objective of this study was to examine the relationship between (type of) dairy intake and CVD mortality and all-cause mortality in a Dutch population.We examined the relationship between dairy intake and CVD mortality and all-cause mortality in 1956 participants of the Hoorn Study (aged 50–75 years), free of CVD at baseline. Hazard ratios with 95 % CIs were obtained for CVD mortality and all-cause mortality per standard deviation (SD) of the mean increase in dairy intake, with adjustment for age, sex, BMI, smoking, education, total energy intake, alcohol consumption, physical activity, and dietary intakes.During 12.4 years of follow-up, 403 participants died, of whom 116 had a fatal CVD event. Overall dairy intake was not associated with CVD mortality or all-cause mortality. Each SD increase in high-fat dairy intake was associated with a 32 % higher risk of CVD mortality (95 % CI; 7–61 %).In this prospective cohort study, the intake of high-fat dairy products was associated with an increased risk of CVD mortality.
Keywords: Dairy; Cardiovascular diseases; Mortality; Prospective

Water intake and post-exercise cognitive performance: an observational study of long-distance walkers and runners by Martin D. Benefer; Bernard M. Corfe; Jean M. Russell; Richard Short; Margo E. Barker (617-624).
The impact of diet on endurance performance and cognitive function has been extensively researched in controlled settings, but there are limited observational data in field situations. This study examines relationships between nutrient intake and cognitive function following endurance exercise amongst a group of 33 recreational runners and walkers.All participants (mean age of 43.2 years) took part in a long-distance walking event and completed diet diaries to estimate nutrient intake across three-time periods (previous day, breakfast and during the event). Anthropometric measurements were recorded. Cognitive tests, covering word recall, ruler drop and trail making tests (TMT) A and B were conducted pre- and post-exercise. Participants rated their exercise level on a validated scale. Nutrient intake data were summarised using principal components analysis to identify a nutrient intake pattern loaded towards water intake across all time periods. Regression analysis was used to ascertain relationships between water intake component scores and post-exercise cognitive function, controlling for anthropometric measures and exercise metrics (distance, duration and pace).Participants rated their exercise as ‘hard-heavy’ (score 14.4, ±3.2). Scores on the water intake factor were associated with significantly faster TMT A (p = 0.001) and TMT B (p = 0.005) completion times, and a tendency for improved short-term memory (p = 0.090). Water intake scores were not associated with simple reaction time (assessed via the ruler drop test).These data are congruent with experimental research demonstrating a benefit of hydration on cognitive function. Further field research to confirm this relationship, supported with precise measures of body weight, is needed.
Keywords: Cognitive function; Memory; Reaction time; Trail making test; Hydration

Disturbed eating at high altitude: influence of food preferences, acute mountain sickness and satiation hormones by Isabelle Aeberli; Annina Erb; Kerstin Spliethoff; Daniela Meier; Oliver Götze; Heiko Frühauf; Mark Fox; Graham S. Finlayson; Max Gassmann; Kaspar Berneis; Marco Maggiorini; Wolfgang Langhans; Thomas A. Lutz (625-635).
Hypoxia has been shown to reduce energy intake and lead to weight loss, but the underlying mechanisms are unclear. The aim was therefore to assess changes in eating after rapid ascent to 4,559 m and to investigate to what extent hypoxia, acute mountain sickness (AMS), food preferences and satiation hormones influence eating behavior.Participants (n = 23) were studied at near sea level (Zurich (ZH), 446 m) and on two days after rapid ascent to Capanna Margherita (MG) at 4,559 m (MG2 and MG4). Changes in appetite, food preferences and energy intake in an ad libitum meal were assessed. Plasma concentrations of cholecystokinin, peptide tyrosine–tyrosine, gastrin, glucagon and amylin were measured. Peripheral oxygen saturation (SpO2) was monitored, and AMS assessed using the Lake Louis score.Energy intake from the ad libitum meal was reduced on MG2 compared to ZH (643 ± 308 vs. 952 ± 458 kcal, p = 0.001), but was similar to ZH on MG4 (890 ± 298 kcal). Energy intake on all test days was correlated with hunger/satiety scores prior to the meal and AMS scores on MG2 but not with SpO2 on any of the 3 days. Liking for high-fat foods before a meal predicted subsequent energy intake on all days. None of the satiation hormones showed significant differences between the 3 days.Reduced energy intake after rapid ascent to high altitude is associated with AMS severity. This effect was not directly associated with hypoxia or changes in gastrointestinal hormones. Other peripheral and central factors appear to reduce food intake at high altitude.
Keywords: Hypoxia; Dietary intake; Food preferences; High altitude; Acute mountain sickness

Influence of breastfeeding versus formula feeding on lymphocyte subsets in infants at risk of coeliac disease: the PROFICEL study by Tamara Pozo-Rubio; Amalia Capilla; Jorge R. Mujico; Giada de Palma; Ascensión Marcos; Yolanda Sanz; Isabel Polanco; Maria Dolores García-Novo; Gemma Castillejo; Carmen Ribes-Koninckx; Vicente Varea; Francesc Palau; Luis Ortigosa; Luis Peña-Quintana; Esther Nova (637-646).
In addition to genetic risk, environmental factors might influence coeliac disease (CD) development. We sought to assess the effect of the interaction between milk-feeding practices and the HLA-DQ genotype on peripheral lymphocyte subsets and their activation markers in infants at familial risk for CD.170 newborns were classified in 3 different genetic risk groups (high risk, HR; intermediate risk, IR; and low risk, LR) after DQB1 and DQA1 typing. Lymphocyte subsets were studied at the age of 4 months by flow cytometry analysis.79 infants were receiving exclusive breastfeeding (BF) and 91 partial breastfeeding or formula feeding (FF). Regarding genetic risk, 40 infants were classified in HR group, 75 in IR group and 55 in LR group. Two-way ANOVA did not show significant interactions between the type of milk feeding and genetic risk group on the lymphocyte subsets analysed. One-way ANOVA for milk-feeding practice alone showed that the percentage of CD4 + CD25+ cells was significantly higher in BF group than in FF group (BF, 10.92 ± 2.71; FF, 9.94 ± 2.96; p = 0.026), and absolute counts of CD4 + CD38+ cells were significantly higher in FF group than in BF group (FF, 2,881.23 ± 973.48; BF, 2,557.95 ± 977.06; p = 0.038). One-way ANOVA for genetic risk alone showed that absolute counts of NK cells were significantly higher in IR group than HR and LR groups (IR, 539.24 ± 340.63; HR, 405.01 ± 239.53; LR, 419.86 ± 262.85; p = 0.028).Lymphocyte subset profiles in the early stages of life could be modulated by milk-feeding practices and genetic risk separately. Breastfeeding might have a positive immunomodulatory effect on lymphocyte subsets in infants at risk of CD.
Keywords: Lymphocyte subsets; Coeliac disease; Infants; HLA genotype; Breastfeeding; Formula feeding

Fish oil, containing mainly long-chain n-3 polyunsaturated fatty acids (LCn-3PUFA), has been found to acutely stimulate protein synthesis and insulin-mediated glucose metabolism. However, the underlying mechanism and more prolonged effect of fish oil during ageing remain to be determined.Fish oil (EPAX6000; 49.6 % eicosapentaenoic acid, 50.4 % docosahexaenoic acid) or control oil (60 % olive, 40 % soy) supplementation was delivered, via chocolate-derived sweets, to rats for 8 weeks. Throughout the study, food intake and body weight were recorded and body composition was investigated using EchoMRI. During the last 40 min of a 6 h infusion, with labelled dextrose ([U-13C]glucose) and amino acids ([1-13C]phenylalanine), blood samples were collected to assess glucose and phenylalanine kinetics. Soleus and longissimus dorsi muscles were extracted for protein and mRNA analyses.Fish oil had no effect on food intake or body composition. An increased whole-body glucose turnover, mainly accounted for via an increase in endogenous glucose production, was observed with fish oil feeding. No effects on whole-body phenylalanine turnover were observed. In longissimus dorsi, fish oil augmented the phosphorylation of phosphoinositide 3-kinase (PI3K)[Tyr458] (P = 0.04) and 70 kDa ribosomal protein S6 kinase (p70s6k)[Thr389] (P = 0.04). There were no differences in protein kinase B (Akt)[Ser473], mammalian target of rapamycin (mTOR)[Ser2448], protein phosphatase 2A (PP2A) 56 kDa regulatory B subunit γ (PP2A-B56-γ), forkhead box containing proteins O-subclass 3a (FOX03a)[Ser253] or inflammatory markers (Interleukin-6, Interleukin-1 β, tumour necrosis factor-α, and cyclooxygenase-2).Our data suggest that the fish oil may stimulate endogenous glucose production and increase anabolic signalling in ageing rats.
Keywords: Ageing; Fish oil; p70s6k; Metabolism; Insulin sensitivity

Cord serum (CS) phospholipid fatty acid composition is associated with maternal diet during foetal life, and maternal intake of linoleic acid (LA, C18:2ω-6) and α-linolenic acid (LNA, C18:3 ω-3) has been shown to influence the LA and LNA levels in CS. A possible connection between the increased incidence of atopic diseases and increased intake of LA and decreased intake of LNA in the Western world has been proposed.The aim of this study was to explore phospholipid fatty acid proportions and total IgE levels in CS from Swedish children, collected from 1985 to 2005, a period with increasing frequency of allergic diseases in Sweden, and reveal possible changes over time.Phospholipid fatty acids and total IgE antibodies were analysed with gas chromatography and UniCAP® technology, respectively, in 300 CS samples.The proportions of LA and LNA decreased significantly from 1985 to 2005 (p < 0.001 for both). However, the LA/LNA ratio did increase (p < 0.001), revealing a relatively larger decrease in LNA than in LA. No correlations were found between ω-6 and ω-3 fatty acids and total IgE antibodies in CS from newborn children.The LA/LNA ratio increased (p < 0.001) in cord serum samples collected between 1985 and 2005, and no correlations between fatty acids and total IgE were found.
Keywords: Cord blood; Fatty acids; Immunoglobulin E

Anthocyanins in the diet of children and adolescents: intake, sources and trends by Claudia Drossard; Katja Bolzenius; Clemens Kunz; Mathilde Kersting (667-676).
Anthocyanin intake estimations in large cohorts include almost exclusively adults. For the purpose of early dietary prevention, however, it is of great interest to estimate anthocyanin intake of children and adolescents.Anthocyanidin content values from the USDA Database (excluding the values for bananas and nuts) were assigned to foods consumed by 4–18-year-old participants of the DONALD Study. Between 1990 and 2009, 920 participants provided 6,707 3-day weighed dietary records. Intake of anthocyanins (expressed as their aglycones anthocyanidins) and their food sources were determined. For investigating age and time trends in anthocyanidin density (mg/MJ), a polynomial mixed regression model was built.We found the estimated median anthocyanidin intake to be around 6 mg/day, strawberries representing the main source. Anthocyanidin density of the diet was about 0.2 mg/MJ higher in girls than in boys, decreased with age, decreased over time in the first half of the study period and increased over time thereafter.Anthocyanin intake in the young is characterised by differences in anthocyanidin density of the diet between girls and boys and by decreasing density from young childhood to adolescence. Observations in this German study population should be extended by further studies in other countries.
Keywords: Anthocyanin intake; Children; Adolescents; Age and time trends; Banana

Overweight and obesity prevalence and determinants in Italy: an update to 2010 by Silvano Gallus; Anna Odone; Alessandra Lugo; Cristina Bosetti; Paolo Colombo; Piergiorgio Zuccaro; Carlo La Vecchia (677-685).
To provide updated information on trends and determinants of underweight, overweight, and obesity in Italian adults.We considered data from 5 surveys conducted annually between 2006 and 2010, on a total of 14,135 subjects aged 18 years or more (6,834 men and 7,301 women), representative of the Italian adult population, including self-reported information on height and weight.Overall, 3.1 % of the Italian adult population was underweight (body mass index, BMI, <18.5 kg/m2; 0.8 % men, 5.3 % women), 31.8 % overweight (25≤ BMI <30 kg/m2; 39.8 % men, 24.4 % women), and 8.9 % obese (BMI ≥30 kg/m2; 8.5 % men, 9.4 % women). We observed no specific pattern of overweight/obesity across calendar years in men (multivariate prevalence ratios, PR, for 2010 vs 2006: 0.95; p for trend: 0.980) and a non-significant decreased trend in women (PR: 0.92; p for trend: 0.051). Prevalence of overweight/obesity significantly increased with age (PRs for ≥65 vs 18–24 years: 2.01 in men, 2.65 in women), decreased with education (PRs for high vs low education: 0.79 in men, 0.54 in women), and was less frequent in single than in married adults (PRs: 0.85 in men, 0.78 in women). Overweight/obesity was significantly more frequent in adults from southern versus northern Italy (PRs: 1.13 in men, 1.32 in women) and in former versus never smokers (PRs: 1.23 in men, 1.19 in women).In Italy, we did not find unfavorable trends in overweight and obesity prevalence across calendar years. However, there are specific subgroups of the population with elevated prevalence of overweight and obesity, mainly adults from southern Italy and less educated ones.
Keywords: BMI; Underweight; Overweight; Obesity; Trend; Italy

Regulation of the exopolysaccharide from an anamorph of Cordyceps sinensis on dendritic cell sarcoma (DCS) cell line by Dan Song; Zhenyue He; Chenhao Wang; Fengjiao Yuan; Ping Dong; Weiyun Zhang (687-694).
Cordyceps sinensis has been regarded as a precious tonic food and herbal medicine in China for thousands of years. The exopolysaccharide (EPS) from an anamorph of Cordyceps sinensis was found to have antitumor immunomodulatory activity. Mature dendritic cells play a role in initiating antitumor immunity, so we try to investigate the effects of EPS on the murine dendritic cell line DCS.Flow cytometry was used to assay the expression levels of cell surface molecules including major histocompatibility complex (MHC)-II, CD40, CD80, and CD86 of DCS cells and their ability to take up antigens. The ability of DCS cells to activate the proliferation of CTLL-2 T cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method. IL-12 and TNF-α levels were detected using ELISA. Western blotting was performed to estimate the levels of phosphorylated Janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), nuclear factor-κB (NF-κB) p65 and p105.EPS increased the expressions of MHC-II, CD40, CD80, and CD86 of DCS cells and up-regulated their ability to take up antigens. EPS also enhanced their ability to activate the proliferation of CTLL-2 T cells. IL-12 and TNF-α secreted from DCS cells were up-regulated after EPS treatment. Furthermore, EPS significantly caused the decline of p-JAK2 and p-STAT3, significantly increased levels of NF-κB p65 in the nucleus and decreased levels of NF-κB p105 in the cytoplasm.EPS may induce DCS cells to exhibit mature characteristics, and the mechanism involved is probably related to the inhibition of the JAK2/STAT3 signal pathway and promotion of the NF-κB signal pathway.
Keywords: Exopolysaccharide; Cordyceps sinensis ; Dendritic cell; JAK2/STAT3; NF-κB

Relationship between vitamin D deficiency, bone remodelling and iron status in iron-deficient young women consuming an iron-fortified food by Ruth Blanco-Rojo; Ana M. Pérez-Granados; Laura Toxqui; Pilar Zazo; Concepción de la Piedra; M. Pilar Vaquero (695-703).
Iron and vitamin D deficiencies are two of the most widespread nutritional disorders in the world. Our aim was to know whether the consumption of an iron-fortified fruit juice modifies bone remodelling and the possible influence of baseline vitamin D status on the recovery of iron status in a group of iron-deficient women.Iron biomarkers, 25-hydroxyvitamin D levels and dietary intake were measured in 123 iron-deficient menstruating women. A subgroup (n = 41) participated in a randomised double-blind placebo-controlled study of 16-weeks during winter. They consumed a placebo fruit juice (P) or iron-fortified fruit juice (F). Dietary intake, 25-hydroxyvitamin D, parathormone (PTH), bone alkaline phosphatase (ALP), aminoterminal telopeptide of collagen I (NTX) and iron biomarkers were determined.Ninety-two per cent of the iron-deficient women were vitamin D deficient or insufficient. Transferrin saturation and 25-hydroxyvitamin D were positively correlated. Iron status improved in F, 25-hydroxyvitamin D decreased in F and P, and PTH, ALP and NTX levels were within the normal range and did not vary. Women with 25-hydroxyvitamin D ≥ 50 nmol/L compared with 25-hydroxyvitamin D < 50 nmol/L showed a higher increase in transferrin saturation (a marker of iron supply to tissues) during iron recovery.The prevalence of vitamin D deficiency or insufficiency is very high in iron-deficient women. The recovery of iron status by consuming an iron-fortified food does not affect 25-hydroxyvitamin D levels; however, the increase in iron supply to tissues is lower if the women also present vitamin D deficiency. Although bone health does not seem to be affected in this group of women, correction of iron and vitamin D deficiencies should be promoted in young women to improve present and future health.
Keywords: Iron deficiency anaemia; 25-hydroxyvitamin D; Bone remodelling; Fortified food; Menstruating women

Glycemic index, glycemic load and endometrial cancer risk: results from the Australian National Endometrial Cancer study and an updated systematic review and meta-analysis by Christina M. Nagle; Catherine M. Olsen; Torukiri I. Ibiebele; Amanda B. Spurdle; Penelope M. Webb (705-715).
The relationship between habitual consumption of foods with a high glycemic index (GI) and/or a diet with a high glycemic load (GL) and risk of endometrial cancer is uncertain, and relatively few studies have investigated these associations. The objectives of this study were to examine the association between GI/GL and risk of endometrial cancer using data from an Australian population-based case–control study and systematically review all the available evidence to quantify the magnitude of the association using meta-analysis.The case–control study included 1,290 women aged 18–79 years with newly diagnosed, histologically confirmed endometrial cancer and 1,436 population controls. Controls were selected to match the expected Australian state of residence and age distribution (in 5-year bands) of cases. For the systematic review, relevant studies were identified by searching PubMed and Embase databases through to July 2011. Random-effects models were used to calculate the summary risk estimates, overall and dose–response.In our case–control study, we observed a modest positive association between high dietary GI (OR 1.43, 95 % CI 1.11–1.83) and risk of endometrial cancer, but no association with high dietary GL (OR 1.15, 95 % CI 0.90–1.48). For the meta-analysis, we collated information from six cohort and two case–control studies, involving a total of 5,569 cases. The pooled OR for the highest versus the lowest intake category of GI was 1.15 (0.95–1.40); however, there was significant heterogeneity (p 0.004) by study design (RR 1.00 [95 % CI 0.87–1.14] for cohort studies and 1.56 [95 % CI 1.21–2.02] for case–control studies). There was no association in the dose–response meta-analysis of GI (RR per 5 unit/day increment of GI 1.00, 95 % CI 0.97–1.03). GL was positively associated with endometrial cancer. The pooled RR for the highest versus the lowest GL intake was 1.21 (95 % CI 1.09–1.33) and 1.06 (95 % CI 1.01–1.11) per 50 unit/day increment of GL in the dose–response meta-analysis.The pooled results from observational studies, including our case–control results, provide evidence of a modest positive association between high GL, but not GI, and endometrial cancer risk.
Keywords: Glycemic load; Glycemic index; Case–control; Dose–response meta-analysis; Endometrial cancer

Protective effect of myo-inositol hexaphosphate (phytate) on bone mass loss in postmenopausal women by Ángel A. López-González; Félix Grases; Nieves Monroy; Bartolome Marí; Mª Teófila Vicente-Herrero; Fernando Tur; Joan Perelló (717-726).
The objective of this paper was to evaluate the relationship between urinary concentrations of InsP6, bone mass loss and risk fracture in postmenopausal women.A total of 157 postmenopausal women were included in the study: 70 had low (≤0.76 μM), 42 intermediate (0.76–1.42 μM) and 45 high (≥1.42 μM) urinary phytate concentrations. Densitometry values for neck were measured at enrollment and after 12 months (lumbar spine and femoral neck), and 10-year risk fracture was calculated using the tool FRAX®.Individuals with low InsP6 levels had significantly greater bone mass loss in the lumbar spine (3.08 ± 0.65 % vs. 0.43 ± 0.55 %) than did those with high phytate levels. Moreover, a significantly greater percentage of women with low than with high InsP6 levels showed more than 2 % of bone mass loss in the lumbar spine (55.6 vs. 20.7 %). The 10-year fracture probability was also significantly higher in the low-phytate group compared to the high-phytate group, both in hip (0.37 ± 0.06 % vs 0.18 ± 0.04 %) and major osteoporotic fracture (2.45 ± 0.24 % vs 1.83 ± 0.11 %).It can be concluded that high urinary phytate concentrations are correlated with reduced bone mass loss in lumbar spine over 12 months and with reduced 10-year probability of hip and major osteoporotic fracture, indicating that increased phytate consumption can prevent development of osteoporosis.
Keywords: Bone mass loss; Bone mineral density; Osteoporosis; Phytate; Risk of fracture

Breast milk fatty acid composition differs between overweight and normal weight women: the STEPS Study by Johanna Mäkelä; Kaisa Linderborg; Harri Niinikoski; Baoru Yang; Hanna Lagström (727-735).
We studied differences in breast milk fatty acid (FA) composition between overweight and normal weight women and the effect of FA composition on children’s cholesterol concentrations at 13 months and growth from birth to 13 months.Samples were collected from lactating women (n = 100) participating in STEPS study at infant’s age of 3 months, and FA composition was analyzed with gas chromatography. Diet of mother was studied with Index of Diet Quality at third trimester of pregnancy and with food frequency questionnaire on sampling day. The children’s weights and heights were collected from hospital records at birth and during study visits at 13 months.Overweight women’s breast milk compared to normal weight women’s breast milk contained higher amount of saturated FAs (46.3 vs. 43.6 %, P = 0.012), lower amount of n-3FAs (2.2 vs. 2.7 %, P = 0.010), lower ratio of unsaturated to saturated FAs (1.1 vs. 1.3, P = 0.008), and higher ratio of n-6 to n-3 FAs (5.7 vs. 4.9, P = 0.031) than those of normal weight women even after adjusting for maternal diet (P < 0.05 for all). Normal weight women adhered more to dietary recommendations during pregnancy, whereas no differences were found in diet at sampling 3 months postpartum. The children’s weight gains correlated with saturated FAs (R = 0.22, P = 0.04) and the ratio of unsaturated to saturated FAs (R = −0.23, P = 0.038) in milk; however, effects diminished after adjusting for total duration of breastfeeding. Milk FA composition was not associated with children’s cholesterol concentrations at 13 months.Breast milk FA composition differed between overweight and normal weight women.
Keywords: Breast milk; Breast feeding; Fatty acids; Infant growth; Nutrition; Obesity

De-alcoholised white and red wines decrease inflammatory markers and NF-κB in atheroma plaques in apoE-deficient mice by Nuria Martínez; Kelly Casós; Paolo Simonetti; María P. Sáiz; Juan J. Moreno; María T. Mitjavila (737-747).
Wine polyphenols attenuate the development of atherosclerosis, which involves an inflammatory process. We studied the beneficial effect of de-alcoholised white and red wines (DWW and DRW, respectively) on the development of atheroma plaques and on the expression of biomarkers.We administered control or de-alcoholised wine-rich diets to apoE-deficient mice for 12 or 20 weeks. We then used optical microscopy or immunofluorescence to examine atherosclerotic lesion development in the thoracic aorta and aortic root and assessed the presence of cytokines and adhesion molecules by qPCR and immunofluorescence in total aorta and aortic root, respectively.Atherosclerotic lesions in thoracic aorta were significantly decreased in mice supplemented with DWW (30 %) and DRW (62 %) for 20 weeks. In addition, the expressions of interferon-γ, interleukin-1β, the monocyte chemoattractant protein-1 and CD68 were reduced by DRW. The adhesion molecule P-selectin, vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 were decreased by 52, 76 and 45 %, respectively, in mice fed DRW for 12 weeks, whereas DWW reduced these parameters in a minor extent. The NF-κB expression in total aorta was significantly decreased in the mice treated with de-alcoholised wines for 12 weeks.DRW is shown to be more effective than DWW on cytokines and adhesion molecule expression, in the early stages of the inflammatory events associated with atherosclerosis development, probably due to the high phenolic content of red wine. Downregulation of NF-κB expression may be involved in the mechanism by which de-alcoholised wines modulate atherosclerosis.
Keywords: Atherosclerosis; Wine polyphenols; Adhesion molecules; Cytokines; Nuclear factor-kappa B; Inflammation

Evidence of altered biochemical composition in the hearts of adult intrauterine growth-restricted rats by Vladislava Zohdi; Bayden R. Wood; James T. Pearson; Keith R. Bambery; M. Jane Black (749-758).
Epidemiological studies clearly link intrauterine growth restriction with increased risk of cardiac disease in adulthood. The mechanisms leading to this increased risk are poorly understood; remodeling of the myocardium is implicated. The aim was to determine the effect of early life growth restriction on the biochemical composition of the left ventricular myocardium in adult rats.Wistar Kyoto dams were fed either a low protein diet (LPD; 8.7 % casein) or normal protein diet (NPD; 20 % casein) during pregnancy and lactation; from weaning, the offspring were fed normal rat chow. At 18 weeks of age, the biochemical composition of the hearts of NPD control (n = 9) and LPD intrauterine growth-restricted (n = 7) offspring was analyzed using Fourier Transform Infrared (FTIR) micro-spectroscopy.Body weights at postnatal day 4 were significantly lower and remained lower throughout the experimental period in the LPD offspring compared to controls. FTIR analysis of the infrared absorption spectra across the whole “fingerprint” region (1,800–950 cm−1) demonstrated wider variation in absorbance intensity in the LPD group compared to controls. In particular, there were marked differences detected in the protein (1,540 cm−1), lipid (1,455 and 1,388 cm−1), proteoglycan (1,228 cm−1) and carbohydrate (1,038 cm−1) bands, indicating increased lipid, proteoglycan and carbohydrate content in the growth-restricted myocardium.In conclusion, changes in the biochemical composition of the myocardium provide a likely mechanism for the increased vulnerability to cardiovascular disease in offspring that were growth restricted in early life.
Keywords: Early life growth restriction; Fourier transform infrared (FTIR) micro-spectroscopy; Cardiac remodeling; Fetal programming

Maternal high-fat diet is associated with altered pancreatic remodelling in mice offspring by Bianca Martins Gregorio; Vanessa Souza-Mello; Carlos Alberto Mandarim-de-Lacerda; Marcia Barbosa Aguila (759-769).
To investigate whether a maternal high-fat diet (HF) during pregnancy and/or suckling periods predisposes adult C57BL/6 mice offspring to morphological pancreatic modifications.Male pups were divided into 5 groups: SC (standard chow)—from dams fed SC during gestation and lactation, maintaining an SC diet from postweaning to adulthood; G—from dams fed HF diets during gestation; L—from dams fed HF diets during lactation; GL—from dams fed HF diets during gestation and lactation; and GL/HF—from dams fed HF diets during gestation and lactation, maintaining an HF diet from postweaning to adulthood. We analysed body mass (BM), plasma insulin, pancreas and adipose tissue structures.During the entire experiment, the SC group had the lowest BM. However, GL/HF offspring were heavier than the other groups. This weight gain was also accompanied by adipocyte hypertrophy. At 3 months, G offspring showed an increased insulin levels and impairment in carbohydrates metabolism. Furthermore, pancreatic islets were hypertrophied in G, GL and GL/HF offspring in comparison with SC offspring.HF diet administration during the gestation period is more harmful than during the lactation period, exerting deleterious effects on pancreatic morphology in addition to larger fat deposits in adult mice offspring.
Keywords: Pancreas remodelling; Maternal high-fat diet; Beta-cell mass; Foetal programming

Low-fat dairy, but not whole-/high-fat dairy, consumption is related with higher serum adiponectin levels in apparently healthy adults by Kaijun Niu; Yoritoshi Kobayashi; Lei Guan; Haruki Monma; Hui Guo; Yufei Cui; Atsushi Otomo; Masahiko Chujo; Ryoichi Nagatomi (771-778).
Although previous studies suggested that higher low-fat dairy consumption lower the risk of type 2 diabetes, the mediating factors are not well understood. Higher baseline adiponectin levels are related with a lower risk of type 2 diabetes. This study evaluated whether low-fat dairy is related with adiponectin in apparently healthy adults.We investigated a cross-sectional (n = 938) and one-year longitudinal (n = 759) relationship between low-fat and whole-/high-fat dairy (both including cow’s milk and yogurt) and adiponectin. Dairy consumption was assessed with a validated food frequency questionnaire. Serum adiponectin was measured by using a specific sandwich enzyme-linked immunosorbent assay.In the cross-sectional analysis, the geometric means (95 % confidence intervals [95 % CIs]) of log-transformed adiponectin related with the low-fat dairy categories were 7.27 (6.80–7.77) for the lowest category, 7.67 (7.09–8.31) for the middle category, and 8.40 (7.73–9.13) for the highest category (p < 0.001) after adjustment for potential confounders (including all lifestyle factors). In the longitudinal analysis, repeated-measures ANCOVA adjusted for confounding factors showed a significant time-by-categories (categories of low-fat dairy) interaction in the change of adiponectin. In contrast, no significant relationship was found between the whole-/high-fat dairy categories and adiponectin.This study has shown that higher consumption of low-fat dairy, but not of whole-/high-fat dairy, is related with higher levels of adiponectin and with the change of adiponectin level at the one-year follow-up. These results suggest that the consumption of low-fat dairy may have a beneficial effect on serum adiponectin levels.
Keywords: Low-fat dairy; Adiponectin; Diabetes risk factors; Insulin resistance

Effects of lipoic acid on AMPK and adiponectin in adipose tissue of low- and high-fat-fed rats by Pedro L. Prieto-Hontoria; Patricia Pérez-Matute; Marta Fernández-Galilea; J. Alfredo Martínez; María J. Moreno-Aliaga (779-787).
Lipoic acid (LA) is an antioxidant with antiobesity and antidiabetic properties. Adiponectin is an adipokine with potent anti-inflammatory and insulin-sensitizing properties. AMP-activated protein kinase (AMPK) is a key enzyme involved in cellular energy homeostasis. Activation of AMPK has been considered as a target to reverse the metabolic abnormalities associated with obesity and type 2 diabetes.The aim of this study was to determine the effects of LA on AMPK phosphorylation and adiponectin production in adipose tissue of low-fat (control diet) and high-fat diet-fed rats.Dietary supplementation with LA reduced body weight and adiposity in control and high-fat-fed rats. LA also reduced basal hyperinsulinemia as well as the homeostasis model assessment (HOMA) levels, an index of insulin resistance, in high-fat-fed rats, which was in part independent of their food intake lowering actions. Furthermore, AMPK phosphorylation was increased in white adipose tissue (WAT) from LA-treated rats as compared with pair-fed animals. Dietary supplementation with LA also upregulated adiponectin gene expression in WAT, while a negative correlation between adiposity-corrected adiponectin levels and HOMA index was found. Our present data suggest that the ability of LA supplementation to prevent insulin resistance in high-fat diet-fed rats might be related in part to the stimulation of AMPK and adiponectin in WAT.
Keywords: Lipoic acid; Obesity; Adiponectin; AMPK

Water-soluble rice bran enzymatic extract attenuates dyslipidemia, hypertension and insulin resistance in obese Zucker rats by Maria L. Justo; Rosalia Rodriguez–Rodriguez; Carmen M. Claro; Maria Alvarez de Sotomayor; Juan Parrado; Maria D. Herrera (789-797).
Rice bran enzymatic extract (RBEE) has advantages compared to the original rice bran or its oils including water solubility, lack of rancidity and increased content in high nutritional proteins and nutraceutical compounds, particularly phytosterols, γ-oryzanol and tocols. Our aim was to determine the beneficial effects of RBEE in the pathogenesis of metabolic syndrome in obese Zucker rats.Obese Zucker rats and their lean littermates were fed a 1 and 5 % RBEE-supplemented diet (O1, O5, L1 and L5). Simultaneously, obese and lean Zucker rats, fed a standard diet, were used as controls (OC and LC, respectively). Body weight, food and water intake, and systolic blood pressure were weekly evaluated. After treatment, biochemical assays of serum glucose, insulin, triglycerides (TG), total cholesterol (TC), non-esterified fatty acids (NEFA), adiponectin and nitrates (NO(x)) were determined.RBEE treatment reduced circulating levels of TG and TC, whereas increased HDL-cholesterol without altering NEFA values in obese rats. The extract also induced a significant dose-dependent reduction of hypertension linked to obesity. RBEE of 5 % improved insulin resistance and subsequently reduced HOMA-IR index without altering serum glucose levels. Obese animals treated with RBEE showed partial restoration of adiponectin levels and a significant attenuation of pro-inflammatory values of NO(x).These findings evidence the nutraceutical properties of RBEE against the pathogenesis of metabolic syndrome by attenuating dyslipidemia, hypertension and insulin resistance as well as by restoring hypoadiponectinemia associated to obesity.
Keywords: Rice bran; Metabolic syndrome; Dyslipidemia; Hypertension; Insulin resistance; Obese Zucker rats

Evaluation of gastric processing and duodenal digestion of starch in six cereal meals on the associated glycaemic response using an adult fasted dynamic gastric model by Simon Ballance; Stefan Sahlstrøm; Per Lea; Nina E. Nagy; Petter V. Andersen; Tzvetelin Dessev; Sarah Hull; Maria Vardakou; Richard Faulks (799-812).
To identify the key parameters involved in cereal starch digestion and associated glycaemic response by the utilisation of a dynamic gastro-duodenal digestion model.Potential plasma glucose loading curves for each meal were calculated and fitted to an exponential function. The area under the curve (AUC) from 0 to 120 min and total digestible starch was used to calculate an in vitro glycaemic index (GI) value normalised against white bread. Microscopy was additionally used to examine cereal samples collected in vitro at different stages of gastric and duodenal digestion.Where in vivo GI data were available (4 out of 6 cereal meals) no significant difference was observed between these values and the corresponding calculated in vitro GI value.It is possible to simulate an in vivo glycaemic response for cereals when the gastric emptying rate (duodenal loading) and kinetics of digestible starch hydrolysis in the duodenum are known.
Keywords: Starch; Glycaemic index; Barley; Oat

Low-protein diet improves blood and urinary glucose levels and renal manifestations of diabetes in C57BLKS-db/db mice by Emi Arimura; Masahisa Horiuchi; Hiroaki Kawaguchi; Noriaki Miyoshi; Kohji Aoyama; Toru Takeuchi (813-824).
Dietary protein content is related clinically to the development of diabetic nephropathy. Here, we investigated how dietary protein content (12–24 % energy) within the range used by humans affected renal manifestations including the expressions of genes involved in the renin-angiotensin (RA) system in control and diabetic mice. Moreover, we examined the effects of dietary protein content on HbA1c and urinary glucose.Control (CT) and leptin receptor-deficient obese (db) mice, 5 weeks old, were fed the diets below. Under ad libitum conditions, mice were fed 12, 18, and 24 % energy from protein (L-, M-, and H-diets) for 8 weeks. Under pair-feeding conditions, db mice were supplied H-diet (db-Hp) to the equivalent energy to that consumed by db-L mice. Renal manifestations and values related to glucose and insulin were examined biochemically and pathologically.Under ad libitum conditions, db mice consumed food and water dose dependently of the dietary protein content, although they were consumed similarly by CT mice. CT-L mice showed lower urinary albumin and kidney weight, in association with lower mRNA levels of angiotensinogen and renin, than CT-H mice. Under pair-feeding conditions, db-L mice showed a lower ratio of kidney/body weight, HbA1C, and urinary glucose, and a higher β-cell distribution rate in the pancreas than db-Hp mice.Low-protein intake in the range used by humans may relieve renal manifestations through the suppressed expression of genes in the renal RA system of CT mice. On the other hand, in db mice, low-protein intake improved hyperglycemia and the renal manifestations of diabetes.
Keywords: Animal model; Diet therapy; Insulin insufficiency; Leptin receptor; Renin-angiotensin system

Waist circumference and the risk of hypertension and prediabetes among Filipino women by Delia B. Carba; Isabelita N. Bas; Socorro A. Gultiano; Nanette R. Lee; Linda S. Adair (825-832).
To examine waist circumference as a risk factor for having hypertension only, impaired fasting glucose only, or both hypertension and impaired fasting glucose, and assess whether the associations vary according to overweight status. Furthermore, optimal cut-offs for waist circumference in overweight women and non-overweight women were explored.Data from 1,871 women aged 35–68 years in the 2005 Cebu Longitudinal Health and Nutrition Survey were used. Multinomial logistic regressions were used to model how waist circumference influenced the likelihood of having the three illness categories compared to having neither condition. Waist circumference cut-offs were explored using receiver operating characteristic analysis.Adjusted for age and other confounders, each cm increase in waist circumference increased the odds of hypertension by 5 % for non-overweight women and 3 % for overweight women; impaired fasting glucose by 9 and 3 % for non-overweight and overweight women, respectively; and hypertension and impaired fasting glucose by 17 % among non-overweight versus 9 % for overweight women. Waist circumference cut-offs for non-overweight women were lower than overweight women.Waist circumference was significantly associated with impaired fasting glucose and both hypertension and impaired fasting glucose, and the associations vary by overweight status.
Keywords: Waist circumference; Overweight; Hypertension; Prediabetes; Filipino women

Characterization of microbial metabolism of Syrah grape products in an in vitro colon model using targeted and non-targeted analytical approaches by Anna-Marja Aura; Ismo Mattila; Tuulia Hyötyläinen; Peddinti Gopalacharyulu; Veronique Cheynier; Jean-Marc Souquet; Magali Bes; Carine Le Bourvellec; Sylvain Guyot; Matej Orešič (833-846).
Syrah red grapes are used in the production of tannin-rich red wines. Tannins are high molecular weight molecules, proanthocyanidins (PAs), and poorly absorbed in the upper intestine. In this study, gut microbial metabolism of Syrah grape phenolic compounds was investigated.Syrah grape pericarp was subjected to an enzymatic in vitro digestion model, and red wine and grape skin PA fraction were prepared. Microbial conversion was screened using an in vitro colon model with faecal microbiota, by measurement of short-chain fatty acids by gas chromatography (GC) and microbial phenolic metabolites using GC with mass detection (GC–MS). Red wine metabolites were further profiled using two-dimensional GC mass spectrometry (GCxGC-TOFMS). In addition, the effect of PA structure and dose on conversion efficiency was investigated by GC–MS.Red wine exhibited a higher degree of C1–C3 phenolic acid formation than PA fraction or grape pericarp powders. Hydroxyphenyl valeric acid (flavanols and PAs as precursors) and 3,5-dimethoxy-4-hydroxybenzoic acid (anthocyanin as a precursor) were identified from the red wine metabolite profile. In the absence of native grape pericarp or red wine matrix, the isolated PAs were found to be effective in the dose-dependent inhibition of microbial conversions and short-chain fatty acid formation.Metabolite profiling was complementary to targeted analysis. The identified metabolites had biological relevance, because the structures of the metabolites resembled fragments of their grape phenolic precursors or were in agreement with literature data.
Keywords: Syrah grape; Red wine; Proanthocyanidins; In vitro colon conversions; Phenolic compounds; Short-chain fatty acids; Metabolite profiling

Changes in cardiac energy metabolic pathways in overweighed rats fed a high-fat diet by Javier Modrego; Natalia de las Heras; Jose J. Zamorano-León; Petra J. Mateos-Cáceres; Beatriz Martín-Fernández; Maria Valero-Muñoz; Vicente Lahera; Antonio J. López-Farré (847-856).
Heart produces ATP through long-chain fatty acids beta oxidation.To analyze whether in ventricular myocardium, high-fat diet may modify the expression of proteins associated with energy metabolism before myocardial function was affected.Wistar Kyoto rats were divided into two groups: (a) rats fed standard diet (control; n = 6) and (b) rats fed high-fat diet (HFD; n = 6). Proteins from left ventricles were analyzed by two-dimensional electrophoresis, mass spectrometry and Western blotting.Rats fed with HFD showed higher body weight, insulin, glucose, leptin and total cholesterol plasma levels as compared with those fed with standard diet. However, myocardial functional parameters were not different between them. The protein expression of 3-ketoacyl-CoA thiolase, acyl-CoA hydrolase mitochondrial precursor and enoyl-CoA hydratase, three long-chain fatty acid β-oxidation-related enzymes, and carnitine-O-palmitoyltransferase I was significantly higher in left ventricles from HFD rats. Protein expression of triosephosphate isomerase was higher in left ventricles from HFD rats than in those from control. Two α/β-enolase isotypes and glyceraldehyde-3-phosphate isomerase were significantly increased in HFD rats as compared with control. Pyruvate and lactate contents were similar in HFD and control groups. Expression of proteins associated with Krebs cycle and mitochondrial oxidative phosphorylation was higher in HFD rats.Expression of proteins involved in left ventricle metabolic energy was enhanced before myocardial functionality was affected in rats fed with HFD. These findings may probably indicate higher cardiac energy requirement due to weight increase by HFD.
Keywords: Fat diet; Energy metabolism; Heart; Proteomics

Erratum to: Bias in protein and potassium intake collected with 24-h recalls (EPIC-Soft) is rather comparable across European populations by Sandra P. Crispim; Anouk Geelen; Jeanne H. M. de Vries; Heinz Freisling; Olga W. Souverein; Paul J. M. Hulshof; Marga C. Ocke; Hendriek Boshuizen; Lene F. Andersen; Jiri Ruprich; Willem De Keyzer; Inge Huybrechts; Lionel Lafay; Maria S. de Magistris; Fulvio Ricceri; Rosario Tumino; Vittorio Krogh; H. Bas Bueno-de-Mesquita; Joline W. J. Beulens; Marie-Christine Boutron-Ruault; Androniki Naska; Francesca L. Crowe; Heiner Boeing; Alison McTaggart; Rudolf Kaaks; Pieter van’t Veer; Nadia Slimani (857-858).