European Journal of Nutrition (v.55, #1)

A possible link between hepatic mitochondrial dysfunction and diet-induced insulin resistance by Raffaella Crescenzo; Francesca Bianco; Arianna Mazzoli; Antonia Giacco; Giovanna Liverini; Susanna Iossa (1-6).
Mitochondria are the main cellular sites devoted to ATP production and lipid oxidation. Therefore, the mitochondrial dysfunction could be an important determinant of cellular fate of circulating lipids, that accumulate in the cytoplasm, if they are not oxidized. The ectopic fat accumulation is associated with the development of insulin resistance, and a link between mitochondrial dysfunction and insulin resistance has been proposed.Recent data on the possible link existing between mitochondrial dysfunction in the liver and diet-induced obesity will be summarized, focusing on the three factors that affect the mitochondrial oxidation of metabolic fuels, i.e. organelle number, organelle activity, and energetic efficiency of the mitochondrial machinery in synthesizing ATP. Search in PubMed relevant articles from 2003 to 2014 was conducted, by using query “liver mitochondria and obesity” “hepatic mitochondria and obesity” “liver mitochondria and high fat diet” and “hepatic mitochondria and high fat diet” and including related articles by the same groups.Several works, by using different physiological approaches, have dealt with alteration in mitochondrial function in obesity and diabetes. Most results show that hepatic mitochondrial function is impaired in models of obesity and insulin resistance induced by high-fat or high-fructose feeding.Since mitochondria are the main producers of both cellular energy and free radicals, dysfunctional mitochondria could play an important role in the development of insulin resistance and ectopic fat storage in the liver, thus supporting the emerging idea that mitochondrial dysfunction is closely related to the development of obesity, type 2 diabetes mellitus and non-alcoholic steatohepatitis.
Keywords: Mitochondria; Liver; Insulin; Degree of coupling

Nutrient reference value: non-communicable disease endpoints—a conference report by J. R. Lupton; J. B. Blumberg; M. L’Abbe; M. LeDoux; H. B. Rice; C. von Schacky; A. Yaktine; J. C. Griffiths (1-10).
Nutrition is complex—and seemingly getting more complicated. Most consumers are familiar with “essential nutrients,” e.g., vitamins and minerals, and more recently protein and important amino acids. These essential nutrients have nutrient reference values, referred to as dietary reference intakes (DRIs) developed by consensus committees of scientific experts convened by the Institute of Medicine of the National Academy of Sciences, Engineering, and Medicine and carried out by the Food and Nutrition Board. The DRIs comprise a set of four nutrient-based reverence values, the estimated average requirements, the recommended dietary allowances (RDAs), the adequate intakes and the tolerable upper intake levels for micronutrient intakes and an acceptable macronutrient distribution range for macronutrient intakes. From the RDA, the US Food and Drug Administration (FDA) derives a labeling value called the daily value (DV), which appears on the nutrition label of all foods for sale in the US. The DRI reports do not make recommendations about whether the DV labeling values can be set only for what have been defined to date as “essential nutrients.” For example, the FDA set a labeling value for “dietary fiber” without having the DV. Nutrient reference values—requirements are set by Codex Alimentarius for essential nutrients, and regulatory bodies in many countries use these Codex values in setting national policy for recommended dietary intakes. However, the focus of this conference is not on essential nutrients, but on the “nonessential nutrients,” also termed dietary bioactive components. They can be defined as “Constituents in foods or dietary supplements, other than those needed to meet basic human nutritional needs, which are responsible for changes in health status (Office of Disease Prevention and Health Promotion, Office of Public Health and Science, Department of Health and Human Services in Fed Regist 69:55821–55822, 2004).” Substantial and often persuasive scientific evidence does exist to confirm a relationship between the intake of a specific bioactive constituent and enhanced health conditions or reduced risk of a chronic disease. Further, research on the putative mechanisms of action of various classes of bioactives is supported by national and pan-national government agencies, and academic institutions, as well as functional food and dietary supplement manufacturers. Consumers are becoming educated and are seeking to purchase products containing bioactives, yet there is no evaluative process in place to let the public know how strong the science is behind the benefits or the quantitative amounts needed to achieve these beneficial health effects or to avoid exceeding the upper level (UL). When one lacks an essential nutrient, overt deficiency with concomitant physiological determents and eventually death are expected. The absence of bioactive substances from the diet results in suboptimal health, e.g., poor cellular and/or physiological function, which is relative and not absolute. Regrettably at this time, there is no DRI process to evaluate bioactives, although a recent workshop convened by the National Institutes of Health (Options for Consideration of Chronic Disease Endpoints for Dietary Reference Intakes (DRIs); March 10–11, 2015; http://health.gov/dietaryguidelines/dri/ ) did explore the process to develop DVs for nutrients, the lack of which result in increased risk of chronic disease (non-communicable disease) endpoints. A final report is expected soon. This conference (CRN-International Scientific Symposium; “Nutrient Reference Value—Non-Communicable Disease (NRV-NCD) Endpoints,” 20 November in Kronberg, Germany; http://www.crn-i.ch/2015symposium/ ) explores concepts related to the Codex NRV process, the public health opportunities in setting NRVs for bioactive constituents, and further research and details on the specific class of bioactives, n-3 long-chain polyunsaturated fatty acids (also termed omega-3 fatty acids) and their constituents, specifically docosahexaenoic acid and eicosapentaenoic acid.
Keywords: Bioactives; Nutrient reference values; nonessential nutrients; Adequate intake; n-3 long-chain polyunsaturated fatty acids; Omega-3 fatty acids; Docosahexaenoic acid (DHA); Eicosapentaenoic acid (EPA)

Consumption of soft drinks and juices and risk of liver and biliary tract cancers in a European cohort by Magdalena Stepien; Talita Duarte-Salles; Veronika Fedirko; Antonia Trichopoulou; Pagona Lagiou; Christina Bamia; Kim Overvad; Anne Tjønneland; Louise Hansen; Marie-Christine Boutron-Ruault; Guy Fagherazzi; Gianluca Severi; Tilman Kühn; Rudolf Kaaks; Krasimira Aleksandrova; Heiner Boeing; Eleni Klinaki; Domenico Palli; Sara Grioni; Salvatore Panico; Rosario Tumino; Alessio Naccarati; H. Bas Bueno-de-Mesquita; Petra H. Peeters; Guri Skeie; Elisabete Weiderpass; Christine L. Parr; José Ramón Quirós; Genevieve Buckland; Esther Molina-Montes; Pilar Amiano; Maria-Dolores Chirlaque; Eva Ardanaz; Emily Sonestedt; Ulrika Ericson; Maria Wennberg; Lena Maria Nilsson; Kay-Tee Khaw; Nick Wareham; Kathryn E. Bradbury; Heather A. Ward; Isabelle Romieu; Mazda Jenab (7-20).
The aim of the study was to assess associations between intake of combined soft drinks (sugar sweetened and artificially sweetened) and fruit and vegetable juices and the risk of hepatocellular carcinoma (HCC), intrahepatic bile duct (IHBC) and biliary tract cancers (GBTC) using data from the European Prospective Investigation into Cancer and Nutrition cohort of 477,206 participants from 10 European countries.After 11.4 years of follow-up, 191 HCC, 66 IHBC and 236 GBTC cases were identified. Hazard ratios and 95 % confidence intervals (HR; 95 % CI) were estimated with Cox regression models with multivariable adjustment (baseline total energy intake, alcohol consumption and intake pattern, body mass index, physical activity, level of educational attainment and self-reported diabetes status).No risk associations were observed for IHBC or GBTC. Combined soft drinks consumption of >6 servings/week was positively associated with HCC risk: HR 1.83; 95 % CI 1.11–3.02, p trend = 0.01 versus non-consumers. In sub-group analyses available for 91 % of the cohort artificially sweetened soft drinks increased HCC risk by 6 % per 1 serving increment (HR 1.06, 95 % CI 1.03–1.09, n cases = 101); for sugar-sweetened soft drinks, this association was null (HR 1.00, 95 % CI 0.95–1.06; n cases = 127, p heterogeneity = 0.07). Juice consumption was not associated with HCC risk, except at very low intakes (<1 serving/week: HR 0.60; 95 % CI 0.38–0.95; p trend = 0.02 vs. non-consumers).Daily intake of combined soft drinks is positively associated with HCC, but a differential association between sugar and artificially sweetened cannot be discounted. This study provides some insight into possible associations of HCC with sugary drinks intake. Further exploration in other settings is required.
Keywords: Hepatocellular carcinoma; Biliary tract cancers; Soft drink; Fruit and vegetable juice; Prospective cohort

Gestational dietary patterns are not associated with blood pressure changes during pregnancy and early postpartum in a Brazilian prospective cohort by Ilana Eshriqui; Ana Amélia Freitas Vilela; Fernanda Rebelo; Dayana Rodrigues Farias; Maria Beatriz Trindade Castro; Gilberto Kac (21-32).
To identify gestational dietary patterns and evaluate the association between these patterns and the blood pressure (BP) rate of change during pregnancy and the postpartum.Prospective cohort study composed of 191 healthy pregnant women. Systolic BP (SBP) and diastolic BP (DBP) were obtained at the 5th–13th, 20th–26th, 30th–36th gestational weeks, and with 30–45 days postpartum. A food frequency questionnaire administered at the 30th–36th gestational week was used to measure dietary intake during pregnancy. Principal component analysis was performed to identify the dietary patterns. A longitudinal linear mixed-effects regression model was used to evaluate the association between the dietary patterns and BP (adjusted for time elapsed after conception and the women’s age, education, parity, body mass index and total energy intake).Three gestational dietary patterns were identified: healthy, common-Brazilian and processed. SBP/DBP mean values (SD) were 110.1 (9.0)/66.9 (7.5), 108.7 (9.0)/64.9 (6.7), 111.3 (9.2)/67.0 (6.9) and 115.0 (10.7)/73.7 (8.6) mmHg at the first, second and third gestational trimesters and postpartum, respectively. Women with higher/lower adherence to the processed pattern presented SBP of 117.9 and 113.0 mmHg (P = 0.037), respectively, during postpartum. No association was found between any of the three dietary patterns and SBP in the multiple longitudinal linear regression models, whereas 1 SD increase in the common-Brazilian pattern was associated with a small change of DBP (β = 0.0006; 95 % CI 4.66e-06, 0.001; P = 0.048).The three dietary patterns identified revealed no association with changes of SBP and DBP levels during pregnancy and at early postpartum in this sample of healthy Brazilian women.
Keywords: Food consumption; Dietary pattern; Blood pressure; Pregnancy; Cohort study

Dehydration affects cardiovascular nitric oxide synthases and caveolins in growing rats by Vanina A. Netti; Agustina N. Iovane; Mariana C. Vatrella; Natalia D. Magnani; Pablo A. Evelson; Elsa Zotta; Andrea L. Fellet; Ana María Balaszczuk (33-43).
During the postnatal stage, cardiovascular nitric oxide (NO) system and caveolins (cav) may be regulated differentially in response to hypovolemic state induced by water restriction. Our aim was to examine the effects of water restriction on NO synthases (NOS) and cav in the atria, ventricle and aorta of growing rats.Male Sprague–Dawley rats aged 25 and 50 days were divided into (n = 15): WR: water restriction 3 days; WAL: water ad libitum 3 days. Systolic blood pressure, NOS activity and NOS/cav protein levels were measured.Dehydration induced a larger increase in SBP in WR25 group. Ventricular NOS activity, endothelial NOS (eNOS) and neuronal isoform (nNOS) of WR25 pups were increased, and both cav were decreased. In the WR50 group, NOS activity remained unchanged. In the atria, NOS activity, eNOS and nNOS decreased in WR25 associated with increased cav-1; in the WR50 group, NOS activity was increased without changes in NOS isoforms. In the aorta of WR25, NOS activity and inducible NOS (iNOS) were decreased; NOS activity was unchanged in WR50, despite the decreased levels of eNOS and increased iNOS, cav-1 and cav-3.NO system adjustments in cardiovascular system under osmotic stress in vivo depend on postnatal age, being eNOS and nNOS, the isoforms that determine NOS activity in cardiac tissue in 25-day-old pups. Changes in cav abundance during hypovolemic state may contribute to age-related NO production.
Keywords: Nitric oxide; Caveolins; Water restriction; Cardiovascular system; Postnatal growth

Low vitamin C values are linked with decreased physical performance and increased oxidative stress: reversal by vitamin C supplementation by Vassilis Paschalis; Anastasios A. Theodorou; Antonios Kyparos; Konstantina Dipla; Andreas Zafeiridis; George Panayiotou; Ioannis S. Vrabas; Michalis G. Nikolaidis (45-53).
It has been suggested that part of the failure of antioxidant supplementation to reduce oxidative stress and promote health is that it has been administered in humans with normal levels of antioxidants.To test this hypothesis, we screened 100 males for vitamin C baseline values in blood. Subsequently, the 10 individuals with the lowest and the 10 with the highest vitamin C values were assigned in two groups. Using a placebo-controlled crossover design, the 20 selected subjects performed aerobic exercise to exhaustion (oxidant stimulus) before and after vitamin C supplementation for 30 days.The low vitamin C group had lower VO2max values than the high vitamin C group. Vitamin C supplementation in this group marginally increased VO2max. Baseline concentration of F2-isoprostanes and protein carbonyls was higher in the low vitamin C group compared to the high vitamin C group. Vitamin C supplementation decreased the baseline concentration of F2-isoprostanes and protein carbonyls in both groups, yet the decrease was greater in the low vitamin C group. Before vitamin C supplementation, F2-isoprostanes and protein carbonyls were increased to a greater extent after exercise in the high vitamin C group compared to the low vitamin C group. Interestingly, after vitamin C supplementation, this difference was narrowed.We show for the first time that low vitamin C concentration is linked with decreased physical performance and increased oxidative stress and that vitamin C supplementation decreases oxidative stress and might increase exercise performance only in those with low initial concentration of vitamin C.
Keywords: Antioxidants; Exercise; Oxidative stress; Physical performance; Supplementation

Effect of low-dose selenium on thyroid autoimmunity and thyroid function in UK pregnant women with mild-to-moderate iodine deficiency by Jinyuan Mao; Victor J. Pop; Sarah C. Bath; Huib L. Vader; Christopher W. G. Redman; Margaret P. Rayman (55-61).
Selenium is an essential trace mineral and a component of selenoproteins that are involved in the production of thyroid hormones and in regulating the immune response. We aimed to explore the effect of low-dose selenium supplementation on thyroid peroxidase antibody (TPO-Ab) concentration and thyroid function in pregnant women from a mild-to-moderate iodine-deficient population.Samples and data were from a secondary analysis of Selenium in PRegnancy INTervention (SPRINT), a double-blind, randomized, placebo-controlled study that recruited 230 women with singleton pregnancies from a UK antenatal clinic at 12 weeks of gestation. Women were randomized to receive 60 µg/day selenium or placebo until delivery. Serum thyroid peroxidase antibodies (TPO-Ab), thyrotropin (TSH) and free thyroxine (FT4) were measured at 12, 20 and 35 weeks and thyroglobulin antibodies (Tg-Ab) at 12 weeks.93.5 % of participants completed the study. Se supplementation had no more effect than placebo in decreasing TPO-Ab concentration or the prevalence of TPO-Ab positivity during the course of pregnancy. In women who were either TPO-Ab or Tg-Ab negative at baseline (Thy-Ab−ve), TSH increased and FT4 decreased significantly throughout gestation (P < 0.001), with no difference between treatment groups. In women who were Thy-Ab+ve at baseline, TSH tended to decrease and was lower than placebo at 35 weeks (P = 0.050). FT4 fell more on Se than placebo supplementation and was significantly lower at 35 weeks (P = 0.029).Low-dose selenium supplementation in pregnant women with mild-to-moderate deficiency had no effect on TPO-Ab concentration, but tended to change thyroid function in Thy-Ab+ve women.
Keywords: Selenium; Iodine; Pregnancy; Thyroid autoimmunity; Thyroid peroxidase antibodies; Thyroid function

Evidence suggests that soy foods have chemoprotective properties that may reduce the risk of certain cancers such as breast and prostate cancer. However, data involving gastrointestinal (GI) have been limited, and the evidence remains controversial. This study aims to determine the potential relationship between dietary soy intake and GI cancer risk with an evaluation of the effects of isoflavone as an active soy constituent.Relevant studies were identified after literature search via electronic databases through May 2014. Subgroup analysis for isoflavone intake (studies n = 10) was performed. Covariants including gender types, anatomical subsites and preparation methods were also evaluated. Pooled adjusted odds ratios (ORs) comparing highest and lowest categories of dietary pattern scores were calculated using a random effects model.Twenty-two case–control and 18 cohort studies were included for meta-analysis, which contained a total of 633,476 participants and 13,639 GI cancer cases. The combined OR was calculated as 0.93 (95 % CI 0.87–0.99; p value heterogeneity = 0.01), showing only a slight decrease in risk, the association was stronger for colon cancer (OR 0.92; 95 % CI 0.96–0.99; p value heterogeneity = 0.163) and colorectal cancer (CRC) (OR 0.92; 95 % CI 0.87–0.97; p value heterogeneity = 0.3). Subgroup analysis for isoflavone intake showed a statistically significant risk reduction with a risk estimate of 0.73 (95 % CI 0.59–0.92; p value heterogeneity = 0), and particularly for CRC (OR 0.76; 95 % CI 0.59–0.98; p value heterogeneity = 0).This study provides evidence that soy intake as a food group is only associated with a small reduction in GI cancer risk. Separate analysis for dietary isoflavone intakes suggests a stronger inverse association.
Keywords: Soy; Isoflavone; Gastrointestinal neoplasms; Colon cancer; Meta-analysis

Plantains can be eaten in various forms providing a good opportunity to study the effect of starch type on glycaemic response, and so three products differing in their types of available carbohydrate and contents of resistant starch were tested.Boiled unripe plantain (BUP), boiled unripe plantain crisps (BUPC), ripe raw plantain (RRP) and white bread as reference (all 25 g available carbohydrate portion) were given to ten pre-screened healthy individuals. Postprandial glycaemic responses and glycaemic indices (GI) were measured.Peak blood glucose for BUP, BUPC and RRP was at 45, 45 and 30 min post-meal time, respectively. The peak blood glucose concentrations for BUP, BUPC and RRP (1.8 ± 0.8, 2.3 ± 0.8, 1.9 ± 0.7 mmol/L, n = 10, respectively) reflected the in vitro quantities/types of rapidly available glucose (RAG) in the samples. On the other hand, mean GI ± SEM values obtained for the test products (BUP = 44.9 ± 3.6, BUPC = 55.0 ± 4.2, RRP = 38 ± 4.4, n = 10) were neither significantly different nor directly correlated with RAG.The results show a potential link between RAG and GI, but the correlation is confounded by the presence of other constituents in the plantains.
Keywords: Plantain; Glycaemic index; Glycaemic response; Resistant starch; Available carbohydrates

Early developmental exposure to high fructose intake in rats with NaCl stimulation causes cardiac damage by I. C. Araujo; R. P. Andrade; F. Santos; E. S. Soares; R. Yokota; C. Mostarda; P. Fiorino; K. De Angelis; M. C. Irigoyen; M. Morris; V. Farah (83-91).
Metabolic syndrome (MS) increases the risk of type 2 diabetes and cardiovascular disease. High consumption of fructose is a proposed cause of increased MS, manifested through hypertension, obesity, insulin resistance, and dyslipidemia. High NaCl also increases the risk of CD. The purpose of this study is to evaluate the influence of fructose and sodium on autonomic dysfunction and its relation with CD in MS. Fructose overload was started at weaning and continued through adulthood.Male Wistar rats (21 days) were divided into four groups: Control (C), fructose consumption (10 %, F), NaCl consumption (salt 1 % for the 10 last days, S), and fructose and NaCl (FS), and monitored for 8 weeks. Metabolic evaluations consisted of Lee index, glycemia, insulin and glucose tolerance tests, triglycerides, and total cholesterol measurements. Cardiovascular parameters measured were arterial pressure (AP) and cardiac function performed by echocardiography. They also measured the influence of renin angiotensin (RAS) and autonomic nervous systems by drug blockage with losartan, atropine, and atenolol.Energy analysis showed no change between groups. Fructose overload induced a MS state, confirmed by insulin resistance, glucose intolerance, and dyslipidemia. Fasting glucose was increased in F and FS rat groups compared with C and S groups. AP was higher in F, S, and FS groups in comparison with the C group. The hypotensive response after sympathetic blockade was increased in F, S, and FS versus C. The cardiac vagal tonus was reduced in F and FS animal groups. The intrinsic heart rate was decreased in the FS group (372 ± 9 bpm) compared with the C group (410 ± 13 bpm). The morphometric measurements evaluated through left ventricular diameter during diastole and the left ventricular diameter during systole decreased in the FS group (16 and 26 %, respectively). Diastolic function was reduced in F and FS. The depressor response induced by losartan was increased in the F group in comparison with other groups. However, there was a uniform increase in plasma ACE activity in all treated groups compared with the C group.Data suggest that early exposure to high fructose intake produced marked alterations in metabolic and cardiovascular function. When stimulated by NaCl, the fructose-fed subjects showed further impairment in cardiac function.
Keywords: Cardiovascular risk; Fructose; Metabolic syndrome; Renin angiotensin system; NaCl

Nutritional adequacy according to carbohydrates and fat quality by Ana Sánchez-Tainta; Itziar Zazpe; Maira Bes-Rastrollo; Jordi Salas-Salvadó; Mónica Bullo; José Vicente Sorlí; Dolores Corella; Mª Isabel Covas; Fernando Arós; Mario Gutierrez-Bedmar; Miquel Fiol; F. García de la Corte; Lluis Serra-Majem; Xavier Pinto; Helmut Schröeder; Emilio Ros; M. Carmen López-Sabater; Ramón Estruch; Miguel Angel Martínez-González (93-106).
To investigate the association between carbohydrate quality, fat quality or adherence to the Mediterranean diet and intake adequacy of 19 micronutrients in the PREDIMED (PREvención con DIeta MEDiterránea) trial, a multicenter, randomized, controlled, parallel group and primary prevention trial conducted in Spain. We assessed baseline dietary intake of 6,542 elderly subjects at high cardiovascular risk through a validated food frequency questionnaire (FFQ) and a validated 14-item Mediterranean diet (Med-diet) score. We used a multidimensional carbohydrate quality index (CQI) using four criteria and a fat quality index (FQI) according to the ratio (MUFA + PUFA)/(SFA + TFA). The probability of intake adequacy was calculated comparing the intakes to DRI, and also using the probabilistic approach. Absolute and adjusted probability of having inadequate intake for either ≥6 DRI or ≥8 DRI were estimated to assess nutritional adequacy according to quintiles of each index. The lowest prevalence of inadequate micronutrient intake (≥8 DRI) was found in the highest quintile of CQI or Med-diet score, and in the lowest quintile of FQI (adjusted fold risk: 1.4, 3.4 and 10.2 respectively in comparison with the lowest quintile). P for trend <0.001 in three multivariable models. A higher CQI or Med-Diet score and a lower FQI were significantly associated with a lower fold risk of unmet EAR values.A multidimensional assessment of CQI can be a useful tool to evaluate the quality of carbohydrates. This score and a 14-item Med-diet score were positively related to overall micronutrient adequacy in elderly participants.
Keywords: Carbohydrate quality; Fat quality; Mediterranean diet; Micronutrient adequacy; PREDIMED trial

Effects of Brazil nut consumption on selenium status and cognitive performance in older adults with mild cognitive impairment: a randomized controlled pilot trial by Bárbara Rita Cardoso; Daniel Apolinário; Verônica da Silva Bandeira; Alexandre Leopold Busse; Regina Miksian Magaldi; Wilson Jacob-Filho; Silvia Maria Franciscato Cozzolino (107-116).
Oxidative stress is closely related to cognitive impairment, and the antioxidant system may be a potential therapeutic target to preserve cognitive function in older adults. Selenium plays an important antioxidant role through selenoproteins. This controlled trial aimed to investigate the antioxidant and cognitive effects of the consumption of Brazil nuts, the best selenium food source.We enrolled 31 older adults with mild cognitive impairment (MCI) who were randomly assigned to ingestion of Brazil nuts or to the control group. Participants of the treatment group consumed one Brazil nut daily (estimated 288.75 µg/day) for 6 months. Blood selenium concentrations, erythrocyte glutathione peroxidase (GPx) activity, oxygen radical absorbance capacity, and malondialdehyde were evaluated. Cognitive functions were assessed with the CERAD neuropsychological battery.Eleven participants of the treated group and nine of the control group completed the trial. The mean age of the participants was 77.7 (±5.3) years, 70 % of whom were female. We observed increased selenium levels after the intervention, whereas the control group presented no change. Among the parameters related to the antioxidant system, only erythrocyte GPx activity change was significantly different between the groups (p = 0.006). After 6 months, improvements in verbal fluency (p = 0.007) and constructional praxis (p = 0.031) were significantly greater on the supplemented group when compared with the control group.Our results suggest that the intake of Brazil nut restores selenium deficiency and provides preliminary evidence that Brazil nut consumption can have positive effects on some cognitive functions of older adults with MCI.
Keywords: Brazil nuts; Selenium; Oxidative stress; Mild cognitive impairment

Skeletal muscles respond differently when piglets are offered a diet 30 % deficient in total sulfur amino acid for 10 days by José Alberto Conde-Aguilera; Louis Lefaucheur; Sophie Tesseraud; Yves Mercier; Nathalie Le Floc’h; Jaap van Milgen (117-126).
Although amino acids (AA) are required for growth, little is known about the effect of a deficient AA supply on the composition and the contractile and metabolic properties of skeletal muscles.Protein metabolism, oxidative catabolism, glutathione system, and fiber-type composition of the longissimus (LM), rhomboideus (RM), and semitendinous (SM) muscles were compared between 42-day-old piglets pair-fed for 10 days either with a diet with a 28 % deficient supply of total sulfur AA (TSAA−) or with a diet with a sufficient supply of total sulfur AA (TSAA+).The relative weight, protein mass, and protein synthesis of LM were 10–32 % lower in TSAA− pigs compared with TSAA+ pigs, while RM and SM were not affected by the TSAA supply. The TSAA supply affected the AA composition of muscles. Concentrations of Met and branched-chain AA were, respectively, 7 and 3 % lower in TSAA− pigs compared with TSAA+ pigs. The His concentration was 30 % higher in LM and SM in TSAA− pigs compared with TSAA+ pigs and unaffected in RM. The activity of citrate synthase was 14 % higher in all three muscles of TSAA− pigs. In these pigs, the β-hydroxy-acyl-CoA dehydrogenase activity was 20 % higher in RM compared with TSAA+ pigs while that of lactate dehydrogenase was 21 % lower in LM. Total and reduced glutathione concentrations were more than 70 % greater in RM than in LM or SM, and these concentrations were approximately 10 % lower in TSAA− pigs than in TSAA+ pigs.Results of this study indicate that a TSAA deficiency affects muscle properties in a muscle-dependent manner increasing the oxidative capacity of RM and reducing growth and glycolytic metabolism of LM.
Keywords: Glutathione; Methionine; Muscle amino acid composition; Muscle fiber types; Oxidative catabolism enzymes; Protein synthesis

Effects of total fibre or resistant starch-rich diets within lifestyle intervention in obese prediabetic adults by Margarita S. Dodevska; Sladjana S. Sobajic; Predrag B. Djordjevic; Vesna S. Dimitrijevic-Sreckovic; Vesna V. Spasojevic-Kalimanovska; Brizita I. Djordjevic (127-137).
Starting from the evidence-based health benefits that resistant starch (RS) shows when added to the diet, our aim in this study was to evaluate the effects of increased fibre intake with two different levels of RS coming from regular daily consumed foods on normalization of glycaemia within lifestyle intervention in the population with risk factors for developing diabetes.Study included 47 overweight and obese men and women with disordered glucoregulation and dyslipidaemia, aged between 45–74, divided into RS and Fibre group. Participants were subjected to the lifestyle and dietary intervention with low-fat and high-fibre (>25 g/day) diet for 12 months and were offered two different dietary advices aimed at increasing total fibre intake in Fibre group and at increasing RS intake in RS group.The intake of macronutrients and total fibre was similar between groups at the end of the study, but achieved RS intake was two times higher in the RS group. Decrease in total cholesterol and non-HDL-cholesterol was more pronounced in RS group in comparison with Fibre group (p = 0.010, p = 0.031, respectively), whereas in Fibre group, a more pronounced effect on glucoregulation was observed: significant fall in glycaemia after 2-h oral glucose tolerance test (7.93 vs 6.96 mmol/L, p = 0.034).At the end of the study, RS-rich diet failed to affect glycaemic control in prediabetic obese individuals in contrast to the regular fibre-rich diet, which indicated that fibre profile could be an important determinant of the effect of dietary intervention.
Keywords: Resistant starch; Fibre; Obesity; Impaired glucoregulation

Active Hexose Correlated Compound (AHCC®) is a cultured mushroom extract that is commercially available and promoted for immune support. Available data suggest that AHCC supplementation affects immune cell populations and immune outcomes, including natural killer cell response to infection. The mechanism by which AHCC exerts its effects is not well understood. The present work aimed to characterize the immunomodulatory activity of AHCC in the gut and to study the effects of AHCC on toll-like receptor (TLR) signaling in intestinal epithelial cells (IECs).BALB/c mice were fed AHCC by gavage. In vivo activities were assessed by immunohistochemistry and cytokine production. The effects of AHCC on ex vivo primary cell culture from IECs were examined after challenge with LPS or E. coli alone or in the presence of anti-TLR-2 and TLR-4 blocking antibodies.Feeding AHCC resulted in increased IgA+ cells in the intestine and increased sIgA, IL-10, and IFN-γ in the intestinal fluid. In IECs, contact with AHCC increased IL-6 production but not to the pro-inflammatory level of positive controls, LPS and E. coli. Blocking TLR-2 and TLR-4 reduced the induction of IL-6 by AHCC, suggesting that these innate receptors are involved in generating the immune response of IECs to AHCC.AHCC may play a role in the orchestration of immune response and the maintenance of immune homeostasis in part by priming the TLR-2 and TLR-4 gate at the intestinal epithelium. Such a response is likely due to the recognition of non-pathogenic food-associated molecular patterns (FAMPs) such as those found associated with other mushroom or yeast-derived compounds.
Keywords: AHCC; Toll-like receptor; E. coli ; Infection; Mushroom; Innate; FAMP

CVD-predictive performances of “a body shape index” versus simple anthropometric measures: Tehran lipid and glucose study by Mohammadreza Bozorgmanesh; Mahsa Sardarinia; Farhad Hajsheikholeslami; Fereidoun Azizi; Farzad Hadaegh (147-157).
To examine whether a body shape index (ABSI) calculated by using waist circumference (WC) adjusted for height and weight could improve the predictive performances for cardiovascular disease (CVD) of the Framingham’s general CVD algorithm and to compare its predictive performances with other anthropometric measures.We analyzed data on a 10-year population-based follow-up of 8,248 (4,471 women) individuals aged ≥30 years, free of CVD at baseline. CVD risk was estimated for a 1 SD increment in ABSI, body mass index (BMI), waist-to-hip ratio (WHpR) and waist-to-height ratio (WHtR), by incorporating them, one at a time, into multivariate accelerated failure time models.ABSI was associated with multivariate-adjusted increased risk of incident CVD among both men (1.26, 95 % CI 1.09–1.46) and women (1.17, 1.03–1.32). Among men, for a one-SD increment, ABSI conferred a greater increase in the hazard of CVD [1.26 (1.09–1.46)] than did BMI [1.06 (0.94–1.20)], WC [1.15(1.03–1.28)], WHpR [1.02 (1.01–1.03)] and WHtR [1.16 (1.02–1.31)], and the corresponding figures among women were 1.17 (1.03–1.32), 1.02 (0.90–1.16), 1.11 (0.98–1.27), 1.03 (1.01–1.05) and 1.14 (0.99–1.03), respectively. ABSI as well as other anthropometric measures failed to add to the predictive ability of the Framingham general CVD algorithm either.Although ABSI could not improve the predictability of the Framingham algorithm, it provides more information than other traditional anthropometric measures in settings where information on traditional CVD risk factors are not available, and it can be used as a practical criterion to predict adiposity-related health risks in clinical assessments.
Keywords: CVD prediction; Obesity; ABSI; Anthropometric measures

Mice fed fish oil diet and upregulation of brown adipose tissue thermogenic markers by Thereza Cristina Lonzetti Bargut; Anna Carolina Alves Gomes Silva-e-Silva; Vanessa Souza-Mello; Carlos Alberto Mandarim-de-Lacerda; Marcia Barbosa Aguila (159-169).
Fish oil (FO) elicits diverse beneficial effects. Reduction in or prevention of body mass (BM) gain in animal models may be associated with modulation of brown adipose tissue (BAT). We aimed to evaluate the effects of different high-fat diets with FO on BAT metabolism and thermogenic markers.C57BL/6 male mice (3-month-old) were fed different diets during 8 weeks: standard-chow diet (SC 10 % fat), high-fat lard diet (HF-L 50 % fat), high-fat lard plus FO diet (HF-L+FO 50 % fat), and high-fat FO diet (HF-FO 50 % fat). We evaluated BM and performed an oral glucose tolerance test. At euthanasia, plasma was collected for leptin, and triacylglycerol measurement and interscapular BAT was dissected and stored for molecular analyses.HF-L group showed elevated BM; glucose intolerance associated with diminished TC10 and GLUT4 expressions; hypertriglyceridemia associated with increased CD36 and diminished CPT1 expression; elevated expression of pro-inflammatory cytokines; and reduced PPAR expression. Furthermore, these animals showed hyperleptinemia with increased expression of thermogenic markers (beta3-AR, PGC1alpha, and UCP1). Conversely, HF-L+FO and HF-FO groups showed reduced BM gain with regularization of glucose tolerance and triglyceridemia, GLUT4, TC10, CD36, CPT1, and cytokines expressions. Both groups exhibited elevated PPAR and thermogenic markers expression in a dose-dependent way.FO improves metabolic profile and upregulates thermogenic markers, suggesting an elevated thermogenesis that leads to reduced BM gain.
Keywords: Omega-3 fatty acids; Fish oil; Thermogenesis; Brown adipose tissue; Uncoupling protein 1

Anthocyanins and phenolic acids from a wild blueberry (Vaccinium angustifolium) powder counteract lipid accumulation in THP-1-derived macrophages by Cristian Del Bo’; Yi Cao; Martin Roursgaard; Patrizia Riso; Marisa Porrini; Steffen Loft; Peter Møller (171-182).
Blueberries are a rich source of anthocyanins (ACNs) and phenolic acids (PA), which are hypothesized to protect against development of atherosclerosis. The present study examined the effect of an ACN- and PA-rich fractions, obtained from a wild blueberry powder, on the capacity to counteract lipid accumulation in macrophages derived from monocytic THP-1 cells. In addition, we tested the capacity of pure ACNs and their metabolites to alter lipid accumulation.THP-1-derived macrophages were incubated with fatty acids (500 μM oleic/palmitic acid, 2:1 ratio) and different concentrations (from 0.05 to 10 μg mL−1) of ACN- and PA-rich fractions, pure ACN standards (malvidin, delphinidin and cyanidin 3-glucoside), and metabolites (syringic, gallic and protocatechuic acids). Lipid accumulation was quantified with the fluorescent dye Nile red.Lipid accumulation was reduced at all concentrations of the ACN-rich fraction tested with a maximum reduction at 10 μg mL−1 (−27.4 %; p < 0.0001). The PA-rich fraction significantly reduced the lipid accumulation only at the low concentrations from 0.05 µg mL−1 to 0.3 µg mL−1, with respect to the control with fatty acids. Supplementation with pure ACN compounds (malvidin and delphinidin-3-glucoside and its metabolic products (syringic and gallic acid)) reduced lipid accumulation especially at the low concentrations, while no significant effect was observed after cyanidin-3-glucoside and protocatechuic acid supplementation.The results demonstrated a potential role of both the ACN- and PA-rich fractions and single compounds in the lipid accumulation also at concentrations close to that achievable in vivo.
Keywords: Wild blueberry; Polyphenols; THP-1 macrophages; Lipid accumulation

Effects of increased wholegrain consumption on immune and inflammatory markers in healthy low habitual wholegrain consumers by Antonios Ampatzoglou; Charlotte L. Williams; Kiranjit K. Atwal; Catherine M. Maidens; Alastair B. Ross; Frank Thielecke; Satya S. Jonnalagadda; Orla B. Kennedy; Parveen Yaqoob (183-195).
Wholegrain (WG) consumption is associated with reduced risk of cardiovascular disease, but clinical data on inflammation and immune function is either conflicting or limited. The objective of this study was to assess the impact of increasing WG consumption to at least 80 g/day on markers of inflammation and glucose metabolism and on phenotypic and functional aspects of the immune system, in healthy, middle-aged adults with low habitual WG intake.Subjects consumed a diet high in WG (>80 g/day) or low in WG (<16 g/day, refined grain diet) in a crossover study, with 6-week intervention periods, separated by a 4-week washout. Adherence to the dietary regimes was achieved by dietary advice and provision of a range of food products, with compliance verified by analysis of plasma alkylresorcinols (ARs).On the WG intervention, WG consumption reached 168 g/day (P < 0.001), accompanied by an increase in plasma ARs (P < 0.001) and fibre intake (P < 0.001), without affecting other aspects of dietary intake. On the WG arm, there were trends for lower ex vivo activation of CD4+ T cells and circulating concentrations of IL-10, C-reactive protein, C-peptide, insulin and plasminogen activator inhibitor-1. The percentage of CD4+ central memory T cells and circulating levels of adipsin tended to increase during the WG intervention.Despite the dramatic increase in WG consumption, there were no effects on phenotypic or functional immune parameters, markers of inflammation or metabolic markers.
Keywords: Alkylresorcinols; Fibre; Wholegrain; Immune; Inflammation

Bifidobacterium pseudocatenulatum CECT7765 promotes a TLR2-dependent anti-inflammatory response in intestinal lymphocytes from mice with cirrhosis by Alba Moratalla; Isabel Gómez-Hurtado; Ángela Moya-Pérez; Pedro Zapater; Gloria Peiró; José M. González-Navajas; Eva Maria Gómez Del Pulgar; José Such; Yolanda Sanz; Rubén Francés (197-206).
Intestinal homeostasis plays an important role in bacteria-derived complications in cirrhosis. Intestinal lymphocytes are responsible for immune effector functions and can be modulated by certain probiotics. We evaluate the interaction between Bifidobacterium pseudocatenulatum CECT7765 and intestinal lymphocytes in mice with cirrhosis.Cirrhosis was induced by intragastrical administration of carbon tetrachloride in Balb/C mice. One week prior to laparotomy, animals received B. pseudocatenulatum CECT7765 (107, 109 or 1010 cfu/daily) or placebo. Chemokine receptor and cytokine expression were evaluated in intestinal lymphocytes. Gut permeability was studied by FITC-LPS recovery in vivo. Luminal antigens, inflammation and functional markers were evaluated in liver samples. Bifidobacterium pseudocatenulatum CECT7765 decreased the expression of pro-inflammatory chemokine receptors CCR6, CCR9, CXCR3 and CXCR6 in intestinal lymphocytes from cirrhotic mice in a concentration-dependent manner. The bifidobacterial strain induced a shift towards an anti-inflammatory cytokine profile in this cell subset. B. pseudocatenulatum CECT7765-induced inflammatory modulation was TLR2-mediated, as in vitro TLR2 blockade inhibited the reduction of TNF-alpha and its receptors and the increase of IL-10 and IL-10 receptor secretion. The recovery rate of administered fluorescence-labelled endotoxin was significantly and dose-dependently lowered with the bifidobacterial strain. The reduced intestinal permeability was associated with a decreased burden of bacterial antigens in the liver of mice treated with B. pseudocatenulatum CECT7765. Liver function and inflammation were improved with the use of the bifidobacterial strain at the highest dose tested (1010 cfu). Bifidobacterium pseudocatenulatum CECT7765 improves gut homeostasis and prevents gut-derived complications in experimental chronic liver disease.
Keywords: Cirrhosis; Endotoxin; B. pseudocatenulatum CECT7765; Inflammation; Toll-like receptor 2; Intestinal lymphocytes

Effect of l-carnitine supplementation on the body carnitine pool, skeletal muscle energy metabolism and physical performance in male vegetarians by Katerina Novakova; Oliver Kummer; Jamal Bouitbir; Sonja D. Stoffel; Ulrike Hoerler-Koerner; Michael Bodmer; Paul Roberts; Albert Urwyler; Rolf Ehrsam; Stephan Krähenbühl (207-217).
More than 95 % of the body carnitine is located in skeletal muscle, where it is essential for energy metabolism. Vegetarians ingest less carnitine and carnitine precursors and have lower plasma carnitine concentrations than omnivores. Principle aims of the current study were to assess the plasma and skeletal muscle carnitine content and physical performance of male vegetarians and matched omnivores under basal conditions and after l-carnitine supplementation.Sixteen vegetarians and eight omnivores participated in this interventional study with oral supplementation of 2 g l-carnitine for 12 weeks. Before carnitine supplementation, vegetarians had a 10 % lower plasma carnitine concentration, but maintained skeletal muscle carnitine stores compared to omnivores. Skeletal muscle phosphocreatine, ATP, glycogen and lactate contents were also not different from omnivores. Maximal oxygen uptake (VO2max) and workload (P max) per bodyweight (bicycle spiroergometry) were not significantly different between vegetarians and omnivores. Sub-maximal exercise (75 % VO2max for 1 h) revealed no significant differences between vegetarians and omnivores (respiratory exchange ratio, blood lactate and muscle metabolites). Supplementation with l-carnitine significantly increased the total plasma carnitine concentration (24 % in omnivores, 31 % in vegetarians) and the muscle carnitine content in vegetarians (13 %). Despite this increase, P max and VO2max as well as muscle phosphocreatine, lactate and glycogen were not significantly affected by carnitine administration.Vegetarians have lower plasma carnitine concentrations, but maintained muscle carnitine stores compared to omnivores. Oral l-carnitine supplementation normalizes the plasma carnitine stores and slightly increases the skeletal muscle carnitine content in vegetarians, but without affecting muscle function and energy metabolism.
Keywords: Vegetarians; l-carnitine supplementation; Spiroergometry; Skeletal muscle; Energy metabolism

Previous studies suggested that magnesium (Mg) might protect against atherosclerosis, but data were scarce in an Asian population. We examined the association of Mg levels in serum and urine with carotid intima-media thickness (cIMT) and serum lipids in Chinese adults.This community-based cross-sectional study recruited 2,837 participants aged 40–75 years in Guangzhou, China. General information, lifestyle factors, serum and urinary concentrations of Mg and cardiometabolic factors were determined. The cIMTs of the common carotid artery (CCA) and the carotid bifurcation (BIF) were measured ultrasonographically.The mean (SD) concentration of serum Mg was 0.85 (0.07) mmol/L and median (IQR) for urinary Mg excretion was 2.29 (1.56–3.51) mmol/L. After adjustment for potential covariates, both serum and the urinary concentrations of Mg were inversely associated with CCA-IMT, but not with BIF-IMT. The regression coefficients (standard errors) were −100 (29) µm (total), −86 (34) µm (women) and −117 (52) µm (men) CCA-IMT per 1 mmol/L of serum Mg, and −41 (8) µm (total), −41 (10) µm (women) and −44 (15) µm (men) CCA-IMT per 1 unit of urinary Mg/creatinine (log mmol/mmol) (all p < 0.05), respectively. Higher serum Mg levels were associated with higher total cholesterol, HDLc, LDLc and triglyceride, but lower non-HDLc/HDLc in total population (all p < 0.05). Similar relationships of urinary Mg with lipoproteins were also found in total population (all p < 0.05).Higher levels of serum and urinary Mg are associated with lower CCA-IMTs, and the role of Mg in lipid metabolism needs further investigation.
Keywords: Carotid intima-media thickness; Lipids; Magnesium; Chinese adults; Serum; Urine

Dietary total antioxidant capacity and mortality in the PREDIMED study by P. Henríquez-Sánchez; A. Sánchez-Villegas; C. Ruano-Rodríguez; A. Gea; R. M. Lamuela-Raventós; R. Estruch; J. Salas-Salvadó; M. I. Covas; D. Corella; H. Schröder; M. Gutiérrez-Bedmar; J. M. Santos-Lozano; X. Pintó; F. Arós; M. Fiol; A. Tresserra-Rimbau; E. Ros; M. A. Martínez-González; L. Serra-Majem (227-236).
The aim of the present study was to assess the association between the dietary total antioxidant capacity, the dietary intake of different antioxidants and mortality in a Mediterranean population at high cardiovascular disease risk. A total of 7,447 subjects from the PREDIMED study (multicenter, parallel group, randomized controlled clinical trial), were analyzed treating data as an observational cohort. Different antioxidant vitamin intake and total dietary antioxidant capacity were calculated from a validated 137-item food frequency questionnaire at baseline and updated yearly. Deaths were ascertained through contact with families and general practitioners, review of medical records and consultation of the National Death Index. Cox regression models were fitted to assess the relationship between dietary total antioxidant capacity and mortality. Dietary total antioxidant capacity was estimated using ferric-reducing antioxidant power assays.A total of 319 deaths were recorded after a median follow-up of 4.3 years. Subjects belonging to the upper quintile of antioxidant capacity were younger, ex-smokers, with high educational level, and more active and had higher alcohol intake. Multivariable-adjusted models revealed no statistically significant difference between total dietary antioxidant capacity and mortality (Q5 vs. Q1 ref HR 0.85; 95 % CI 0.60–1.20) neither for the intake of all the vitamins studied.No statistically significant association was found between antioxidant capacity and total mortality in elderly subjects at high cardiovascular risk.
Keywords: Dietary antioxidant capacity; Antioxidant intake; Mortality; PREDIMED

We compared the effects of a eucaloric moderate-fat diet (18 % protein, 36 % fat, and 46 % carbohydrate), a eucaloric low-fat high-carbohydrate diet (18 % protein, 18 % fat, and 64 % carbohydrate), and a low-calorie (33 % reduced) low-fat high-carbohydrate diet on biomarkers of systemic inflammation.We randomly assigned 102 participants (age 21–76 years and BMI 19.2–35.5 kg/m2) to the three different diets for 6 weeks in a parallel design intervention trial. All foods were provided. Ninety-three participants completed all study procedures; 92 were included in the analyses. Endpoints included plasma C-reactive protein (CRP), interleukin-6 (IL-6), soluble tumor necrosis factor receptors I and II (sTNFRI and II), and adiponectin.In the unadjusted primary analyses, none of the endpoints were differentially affected by the dietary interventions despite the significantly greater reductions in body weight and fat mass in participants consuming the low-calorie low-fat diet compared to the eucaloric diets (p < 0.001). When including weight change in the model in secondary analysis, adiponectin tended to be increased with weight loss (time × weight change interaction, p = 0.051). Adjusted for weight change, adiponectin was reduced in the groups consuming the low-fat diets relative to the moderate-fat diet (p = 0.008). No effect of the intervention diets or weight loss on CRP, IL-6, or sTNFRI and II was seen in these secondary analyses.In relatively healthy adults, moderate weight loss had minimal effects on systemic inflammation, and raised plasma adiponectin only modestly. A lower dietary fat and higher carbohydrate content had little impact on measures of systemic inflammation, but reduced adiponectin concentrations compared to a moderate-fat diet. The latter may be of concern given the consistent and strong inverse association of plasma adiponectin with many chronic diseases.
Keywords: Systemic inflammation; Dietary fat; Dietary carbohydrate; Caloric restriction; Adiponectin

FTO gene variation, macronutrient intake and coronary heart disease risk: a gene–diet interaction analysis by Jaana Gustavsson; Kirsten Mehlig; Karin Leander; Christina Berg; Gianluca Tognon; Elisabeth Strandhagen; Lena Björck; Annika Rosengren; Lauren Lissner; Fredrik Nyberg (247-255).
The fat mass and obesity-associated gene (FTO) is related to obesity and coronary heart disease (CHD). We studied interaction between macronutrient intake and FTO in association with CHD risk or body mass index (BMI). The pooled population-based case–control studies, SHEEP and INTERGENE, included 1,381 first-time CHD patients and 4,290 population controls genotyped for FTO rs9939609 (T/A). Diet data were collected in self-administered food frequency questionnaires. Macronutrients were dichotomized into low/high energy percentages (E %) by median levels in controls. Association of FTO genotype (TA/AA vs. TT) with CHD risk was analysed by multiple logistic regression, and with BMI by multiple linear regression. Interaction between FTO and macronutrient was assessed by introducing an interaction term FTO × macronutrient. Interaction on CHD as deviation from additive effects was assessed by calculating relative excess risk due to interaction. No statistically significant interaction was found between FTO genotype and any macronutrient on CHD risk or BMI on either the multiplicative or additive scale. However, FTO genotype (TA/AA vs. TT) was associated with significantly increased CHD risk only in subjects with low E % from fat (OR 1.36, 95 % CI 1.11–1.66) or saturated fatty acids (OR 1.36, 95 % CI 1.10–1.69), or in subjects with high E % from carbohydrate (OR 1.32, 95 % CI 1.07–1.61) or protein (OR 1.41, 95 % CI 1.13–1.75). Mean BMI was 0.3–0.6 kg/m2 higher in control subjects with TA/AA compared to TT, regardless of macronutrient E %.We found no evidence of interactions between FTO genotype and macronutrient intake on CHD risk or BMI.
Keywords: FTO genotype; Coronary heart disease; Macronutrients; Gene–diet interaction

The flavanone isoxanthohumol (IX) has gained attention as antioxidative and chemopreventive agent, but the molecular mechanism of action remains unclear. We investigated effects of this secondary plant compound in vivo using the model organism Caenorhabditis elegans.Adult C. elegans nematodes were incubated with IX, and then, the stress resistance was analysed in the SYTOX assay; lifespan was monitored by touch-provoked movement method, the amount of reactive oxygen species (ROS) was measured in the DCF assay, and the nuclear localisation of the transcription factor DAF-16 was analysed by using a transgenic strain. By the use of a DAF-16 loss-of-function strain, we analysed whether the effects are dependent on DAF-16.IX increases the resistance of the nematode against thermal stress. Additionally, a reduction in ROS in vivo was caused by IX. Since the flavanone only has a marginal radical-scavenging capacity (TEAC assay), we suggest that IX mediates its antioxidative effects indirectly via activation of DAF-16 (homologue to mammalian FOXO proteins). The nuclear translocation of this transcription factor is increased by IX. In the DAF-16-mutated strain, the IX-mediated increase in stress resistance was completely abolished; furthermore, an increased formation of ROS and a reduced lifespan was mediated by IX.IX or a bacterial metabolite of IX causes antioxidative effects as well as an increased stress resistance in C. elegans via activation of DAF-16. The homologous pathway may have implications in the molecular mechanism of IX in mammals.
Keywords: Ageing; Beer; DAF-16; Nutrition; Oxidative stress; Secondary plant compounds

The traditional Japanese dietary pattern and longitudinal changes in cardiovascular disease risk factors in apparently healthy Japanese adults by Kaijun Niu; Haruki Momma; Yoritoshi Kobayashi; Lei Guan; Masahiko Chujo; Atsushi Otomo; Eriko Ouchi; Ryoichi Nagatomi (267-279).
Few epidemiological studies have assessed the relationship between the traditional Japanese dietary pattern and longitudinal changes in cardiovascular disease risk factors among Japanese people. We designed a 3-year longitudinal study of 980 subjects living in Japan to evaluate how the Japanese dietary pattern is related to longitudinal changes in well-recognized risk factors for cardiovascular disease among apparently healthy Japanese adults.Dietary consumption was assessed via a validated food frequency questionnaire. Principal component analysis was used to derive three major dietary patterns—“Japanese,” “sweets-fruits-cooked wheaten food,” and “Izakaya (Japanese Pub)” from 39 food groups.After adjustment for potential confounders, the mean (95 % confidence interval) for the change per year in diastolic blood pressure for men, systolic blood pressure, and diastolic blood pressure for women related to the “Japanese” dietary pattern factor score tertiles were 0.89 (0.10, 1.68), 2.25 (0.19, 4.31), and 0.75 (−1.00, 2.50) for the lowest tertile, 0.77 (−0.02, 1.56), 1.01 (−1.13, 3.15), and 0.44 (−1.38, 2.26) for the middle tertile and − 0.04 (−0.81, 0.72), −0.48 (−2.52, 1.56), and −0.77 (−2.51, 0.96) for the highest tertile (trend P value = 0.03, <0.01, and 0.04, respectively). A significant detrimental relationship was found between the “Izakaya (Japanese Pub)” pattern factor score tertiles and the longitudinal change in serum triglyceride concentration only in men (trend P value = 0.02).Greater adherence to a traditional Japanese diet was independently related to a decreased change every year in diastolic blood pressure in men and women and in systolic blood pressure in women over a 3-year follow-up period. The findings suggest that the “Japanese” dietary pattern appeared to be related to a fall in blood pressure, which might have a beneficial effect on cardiovascular disease. A randomized trial is required to clarify the underlying mechanism.
Keywords: Japanese dietary pattern; Cardiovascular diseases; Japanese adult

Vitamin B12 supplementation during pregnancy and postpartum improves B12 status of both mothers and infants but vaccine response in mothers only: a randomized clinical trial in Bangladesh by Towfida J. Siddiqua; Shaikh M. Ahmad; Khalid B. Ahsan; Mamunur Rashid; Anjan Roy; Syed M. Rahman; Setareh Shahab-Ferdows; Daniela Hampel; Tahmeed Ahmed; Lindsay H. Allen; Rubhana Raqib (281-293).
Poor vitamin B12 (B12) status is associated with adverse outcomes in pregnancy and infancy. Little is known about effects of B12 supplementation on immune function. The present study aimed to evaluate effects of pre- and postnatal B12 supplementation on biomarkers of B12 status and vaccine-specific responses in mothers and infants.In a blinded, placebo-controlled trial, Bangladeshi women (n = 68, age 18–35 years, hemoglobin <110 g/L, 11–14 weeks pregnant) were randomized to receive 250 μg/day B12 or a placebo throughout pregnancy and 3-month postpartum along with 60 mg iron + 400 μg folate. Women were immunized with pandemic influenza A (H1N1) vaccine at 26- to 28-week gestation. Blood from mothers (baseline, 72-h post-delivery, 3-month postpartum), newborns and infants (3-month) was analyzed for hemoglobin, B12, methylmalonic acid (MMA), total homocysteine (tHcy), ferritin and serum transferrin receptor, C-reactive protein (CRP) and alpha-1-acid glycoprotein (AGP). Vitamin B12 was also assessed in breast milk. H1N1-specific antibodies were determined in plasma and colostrum/breast milk.At baseline, 26 % women were B12 deficient (<150 pmol/L), 40 % had marginal status (150–220 pmol/L), 43 % had elevated MMA (>271 nmol/L), and 31 % had elevated tHcy (>10 μmol/L). Supplementation increased B12 in plasma, colostrums and breast milk (p < 0.05) and lowered MMA in neonates, mothers and infants at 3 months (p < 0.05). B12 supplementation significantly increased H1N1-specific IgA responses in plasma and colostrums in mothers and reduced proportion of infants with elevated AGP and CRP compared with placebo.Supplementation with 250 μg/day B12 during pregnancy and lactation substantially improved maternal, infant and breast milk B12 status. Maternal supplementation improved H1N1 vaccine-specific responses in mothers only and may alleviate inflammatory responses in infants.
Keywords: Vitamin B12 supplementation; Methylmalonic acid; Breast milk B12; Vaccine response

Controversy exists regarding whether increasing dairy intake without energy restriction would lead to weight loss. We aimed to compare the potential weight-reducing effects of kefir drink (a probiotic dairy product) and milk in a dairy-rich non-energy-restricted diet in overweight or obese premenopausal women.One hundred and forty-four subjects were assessed for eligibility in this single-center, multi-arm, parallel-group, randomized controlled trial. Of these, seventy-five eligible women aged 25–45 years were randomly assigned to three groups, labeled as control, milk, and kefir, to receive an outpatient dietary regimen for 8 weeks. Subjects in the control group received a diet providing a maintenance level of energy intake, containing 2 servings/day of low-fat dairy products, while those in the milk and kefir groups received a weight maintenance diet, containing 2 additional servings/day (a total of 4 servings/day) of dairy products from low-fat milk or commercial kefir drink, respectively. Anthropometric outcomes including weight, body mass index (BMI), and waist circumference (WC) were measured every 2 weeks.Fifty-eight subjects completed the study. Using analysis of covariance models in the intention-to-treat population (n = 75), we found that at 8 weeks, subjects in the kefir and milk groups had significantly greater reductions in weight, BMI, and WC compared to those in the control group (all p < 0.01). However, no such significant differences were found between the kefir and milk groups.Kefir drink leads to a similar weight loss, compared with milk, in a dairy-rich non-energy-restricted diet in overweight or obese premenopausal women. However, further studies are warranted.
Keywords: Dairy products; Diet; Weight loss; Women; Randomized controlled trial

Vitamin D metabolites and fibroblast growth factor-23 in patients with left ventricular assist device implants: association with stroke and mortality risk by A. Zittermann; M. Morshuis; J. Kuhn; S. Pilz; J. B. Ernst; C. Oezpeker; J. Dreier; C. Knabbe; J. F. Gummert; H. Milting (305-313).
Stroke and mortality risk in patients with left ventricular assist device (LVAD) implants continue to be high. Whether nonclassical cardiovascular risk markers such as vitamin D metabolites and fibroblast growth factor (FGF)-23 contribute to this risk remains to be studied, and this was the objective of our work. In 154 LVAD patients (91 HeartWare and 63 HeartMate II implants), we measured circulating 25-hydroxyvitamin D (25OHD), 1,25-dihydroxyvitamin D3 (1,25[OH]2D3), parathyroid hormone (PTH) and FGF-23 shortly before LVAD implantation and investigated their association with stroke and mortality risk during 1-year follow-up.Of the study cohort, 34.4 and 92.2 %, respectively, had deficient 25OHD (<25 nmol/l) and 1,25(OH)2D3 (<41 pmol/l) values, whereas 42.6 and 98.7 %, respectively, had elevated PTH levels (>6.7 pmol/l) and FGF-23 values above the reference range (100 RU/ml). One-year freedom from stroke was 80.9 %, and 1-year survival was 64.3 %. The multivariable-adjusted hazard ratio of stroke was 2.44 (95 % CI: 1.09–5.45; P = 0.03) for the subgroup of 25OHD levels <25 nmol/l (reference group: 25OHD levels ≥25 nmol/l). The multivariable-adjusted hazard ratio of 1-year mortality was 2.78 (95 % CI: 1.52–5.09; P = 0.001) for patients with 25OHD levels <25 nmol/l compared with patients with 25OHD levels ≥25 nmol/l. PTH, FGF-23 and 1,25(OH)2D3 were not associated with stroke or mortality risk.In LVAD patients, deficient 25OHD levels are independently associated with high stroke and mortality risk. If confirmed in randomized controlled trials, preoperative correction of deficient vitamin D status could be a promising measure to reduce stroke and mortality risk in LVAD patients.
Keywords: Vitamin D; 25-hydroxyvitamin D; 1,25-dihydroxyvitamin D; Fibroblast growth factor-23; Stroke; Mortality

Dietary extra-virgin olive oil prevents inflammatory response and cartilage matrix degradation in murine collagen-induced arthritis by María Angeles Rosillo; Marina Sánchez-Hidalgo; Susana Sánchez-Fidalgo; Marina Aparicio-Soto; Isabel Villegas; Catalina Alarcón-de-la-Lastra (315-325).
Current experimental studies support a beneficial role of extra-virgin olive oil (EVOO) in several inflammatory diseases. The present study was designed to evaluate the effects of dietary EVOO on type II collagen-induced arthritis (CIA) in mice. DBA-1/J mice were randomized in four experimental groups (10 or 15 animals per group): (1) Sham sunflower diet (SO-Sham), (2) CIA sunflower diet (SO-CIA), (3) Sham EVOO diet (EVOO-Sham) and (4) CIA EVOO diet (EVOO–CIA) group. After 6 weeks, arthritis was induced by type II collagen. Mice were sacrified 42 days after first immunization. In addition to macroscopic and histological analyses, serum levels of cartilage olimeric matrix protein (COMP), metalloproteinase-3 (MMP-3) and pro-inflammatory cytokines levels were evaluated by ELISA. The expressions of heme oxygenase-1 (HO-1), nuclear factor E2-related factor 2 (Nrf2), mitogen-activated protein kinases (MAPKs), Janus kinase-signal transducer and activator of transcription (JAK/STAT) and nuclear transcription factor-kappa B (NF-κB) pathways were studied by western blotting.EVOO diet significantly reduced joint edema and cartilage destruction, preventing the arthritis development. Dietary EVOO significantly decreased serum COMP and MMP-3 levels, as well as, the pro-inflammatory cytokines levels (TNF-α, IL-1β and IL-17). Moreover, the activation of JAK/STAT, MAPKs and NF-κB pathways was drastically ameliorated. According to Nrf2 and HO-1, the protein expressions were up-regulated in those mice fed with EVOO.These results support the interest of EVOO as a beneficial functional food to prevent the development of the rheumatoid arthritis (RA).
Keywords: CIA; EVOO; Inflammatory response; Olive oil; Rheumatoid arthritis

Self-reported eating speed in relation to non-alcoholic fatty liver disease in adults by Saehyun Lee; Byung-Joon Ko; Younghoon Gong; Kyungdo Han; Anna Lee; Byoung-Duck Han; Yeo Joon Yoon; Siyoung Park; Jung-Hyun Kim; Christos S. Mantzoros (327-333).
Non-alcoholic fatty liver disease (NAFLD), known to be related to insulin resistance, has been the focus of intensive research efforts due to its increasing prevalence and clinical significance. Rapid eating behavior is another emerging health issue associated with insulin resistance. We aimed to clarify the correlation between self-reported eating speed and NAFLD, both known to be related to insulin resistance.A cross-sectional study was conducted during routine medical checkups on 7,917 consecutively enrolled participants. Anthropometric, biochemical, nutritional, and social parameters were checked. The self-reported eating speed per their usual meal (<5, 5–10, 10–15, and more than 15 min) was recorded by a registered dietitian.The faster eating groups had a higher proportion of NAFLD, and the grade of NAFLD was advanced. After controlling for anthropometric, cardiometabolic, social, and nutritional parameters, the fastest eating group (<5 min) showed an increased risk of NAFLD compared with the lowest eating speed group (≥15 min) both in total [odds ratio (OR) 1.81, 95 % confidence interval (CI) 1.24–2.63] and the participants with BMI < 25 kg/m2 (OR 1.79, 95 % CI 1.22–2.61). As the self-reported eating speed increased, the risk of NAFLD also increased in total and those with BMI < 25 kg/m2 (P for trend <0.001).Fast eating is associated with an increased risk of the presence and grade of NAFLD in Korean adults, especially those with BMI < 25 kg/m2, since presence of overweight or obesity may be overwhelming the effect on NAFLD.
Keywords: Eating behavior; Non-alcoholic fatty liver disease; Insulin resistance; Obesity

Iodine status from childhood to adulthood in females living in North-East Italy: Iodine deficiency is still an issue by Sara Watutantrige Fernando; Elisabetta Cavedon; Davide Nacamulli; Dina Pozza; Andrea Ermolao; Marco Zaccaria; Maria Elisa Girelli; Loris Bertazza; Susi Barollo; Caterina Mian (335-340).
This survey aimed to assess iodine status in a female population at different ages, also investigating their eating habits. We measured urinary iodine concentrations (UIC) in: 634 females at puberty and 361 fertile women in 246 of whom were considered also their children (134 daughters and 120 sons). All subjects completed a food frequency questionnaire.Median UIC decreased from childhood to adulthood (median UIC 107, 77 and 55 μg/l in the young girls, females at puberty and fertile women, respectively). Though using iodized salt improved iodine status in all groups, a significantly higher UIC was only noted in females at puberty. Milk consumption significantly increased UIC at all ages. In mother–child (both daughters and sons) pairs, the children’s median UIC was nearly twice as high as their mothers’ (UIC 115 vs. 57 μg/l). Milk consumption varied significantly: 56 % of the mothers and 76 % of their children drank milk regularly. The children (both daughters and sons) and mothers who drank milk had UIC ≥100 μg/l in 59 and 34 % of cases, respectively, among the pairs who did not drink milk, 44 % of the children and 19 % of the mothers had UIC ≥100 μg/l. On statistical regression, 3.6 % of the variability in the children’s UIC depended on that of their mothers.Dietary iodine status declines from childhood to adulthood in females due to different eating habits. A mild iodine deficiency emerged in women of child-bearing age that could have consequences during pregnancy and lactation.
Keywords: Iodine; Milk; Iodized salt; Childhood; Pregnancy

The combination of resveratrol and quercetin enhances the individual effects of these molecules on triacylglycerol metabolism in white adipose tissue by Noemí Arias; M. Teresa Macarulla; Leixuri Aguirre; Iñaki Milton; María P. Portillo (341-348).
The aim of this study was to analyze whether the combination of resveratrol and quercetin showed additive or synergic effects on body fat accumulation and triacylglycerol metabolism in adipose tissue from rats fed an obesogenic diet.Rats were divided into four dietary groups: a control group and three groups each treated with either resveratrol (15 mg/kg/day; RSV), quercetin (30 mg/kg/day; Q), or both (15 mg resveratrol/kg/day and 30 mg quercetin/kg/day; RSV + Q) for 6 weeks. White adipose tissues from several anatomical locations were dissected. Serum parameters were analyzed by using commercial kits. The activities of fatty acid synthase and heparin-releasable lipoprotein lipase (HR-LPL) were measured using spectrophotometric and fluorimetric methods, and protein expression of acetyl-CoA carboxylase (ACC), adipose tissue triglyceride lipase (ATGL), and hormone-sensitive lipase (HSL) by western blot.The administration of either resveratrol or quercetin separately did not induce significant reductions in adipose tissue weights. By contrast, the combination of both molecules led to a significant reduction in all the fat depots analyzed. The percentage of reduction in each tissue was greater than the calculated additive effect. HR-LPL activity was reduced in RSV and RSV + Q groups. The activity of HSL was not modified. By contrast, ACC was inhibited and ATGL increased only by the combination of both polyphenols.The results obtained demonstrate a synergistic effect between resveratrol and quercetin and suggest that when these molecules are combined, a great number of metabolic pathways involved in adipose tissue triacylglycerol accumulation are affected.
Keywords: Resveratrol; Quercetin; Obesity; White adipose tissue; Rat

Dairy product consumption and risk of type 2 diabetes in an elderly Spanish Mediterranean population at high cardiovascular risk by Andrés Díaz-López; Mònica Bulló; Miguel A. Martínez-González; Dolores Corella; Ramon Estruch; Montserrat Fitó; Enrique Gómez-Gracia; Miquel Fiol; Francisco Javier García de la Corte; Emilio Ros; Nancy Babio; Lluís Serra-Majem; Xavier Pintó; Miguel Ángel Muñoz; Francisco Francés; Pilar Buil-Cosiales; Jordi Salas-Salvadó (349-360).
The possible effects of dairy consumption on diabetes prevention remain controversial. The aim of this study was to investigate the association between the dairy consumption and type 2 diabetes (T2D) risk in an elderly Mediterranean population at high cardiovascular risk.We prospectively followed 3,454 non-diabetic individuals from the PREDIMED study. Dairy consumption was assessed at baseline and yearly using food frequency questionnaires and categorized into total, low-fat, whole-fat, and subgroups: milk, yogurt, cheeses, fermented dairy, concentrated full fat, and processed dairy. Hazard ratios (HRs) were calculated using Cox proportional hazards regression models.During a median follow-up of 4.1 years, we documented 270 incident T2D cases. After multivariate adjustment, total dairy product consumption was inversely associated with T2D risk [0.68 (95 % CI 0.47–0.98); P-trend = .040]. This association appeared to be mainly attributed to low-fat dairy; the multivariate HRs (95 % CIs) comparing the highest versus the lowest tertile consumption were 0.65 (0.45–0.94) for low-fat dairy products and 0.67 (0.46–0.95) for low-fat milk (both P-trend <.05). Total yogurt consumption was associated with a lower T2D risk [HR 0.60 (0.42–0.86); P-trend = .002]. An increased consumption of total low-fat dairy and total yogurt during the follow-up was inversely associated with T2D; HRs were 0.50 (0.29–0.85), 0.44 (0.26–0.75), and 0.55 (0.33–0.93), respectively. Substituting one serving/day of a combination of biscuits and chocolate and whole grain biscuits and homemade pastries for one serving/day of yogurt was associated with a 40 and 45 % lower risk of T2D, respectively. No significant associations were found for the other dairy subgroups (cheese, concentrated full fat, and processed dairy products).A healthy dietary pattern incorporating a high consumption of dairy products and particularly yogurt may be protective against T2D in older adults at high cardiovascular risk.
Keywords: PREDIMED; Type 2 diabetes; Older adults; Dairy; Yogurt

Influence of diet supplementation with green tea extract on drug-metabolizing enzymes in a mouse model of monosodium glutamate-induced obesity by Iva Boušová; Petra Matoušková; Hana Bártíková; Barbora Szotáková; Veronika Hanušová; Veronika Tománková; Eva Anzenbacherová; Barbora Lišková; Pavel Anzenbacher; Lenka Skálová (361-371).
Consumption of dietary supplements with green tea extract (GTE) is popular for weight management, but it may be accompanied by various side effects, including interactions with drugs. The aim of the present in vivo study was to evaluate the effect of defined GTE (Polyphenon 60) in three dosage schemes on insulin, leptin and drug-metabolizing enzymes in obese mice.Experimental obesity was induced by repeated s.c. application of monosodium glutamate to newborn mice. Green tea extract was administered in three dosage schemes in chow diet. The plasmatic levels of insulin and leptin were assayed using enzyme-linked immunosorbent assay. Enzyme activities and mRNA expressions of drug-metabolizing enzymes (totally 13) were analyzed in liver and small intestine using spectrophotometric and HPLC assays and RT-PCR, respectively.GTE-treatment decreased insulin and leptin levels. Eleven enzymes were significantly affected by GTE-treatment. Long-term administration of 0.01 % GTE caused increase in the activity and mRNA level of cytochrome P450 3A4 (CYP3A4) ortholog in the liver as well as in the small intestine. Interestingly, short-term overdose by GTE (0.1 %) had more pronounced effects on enzyme activities and mRNA expressions than long-term overdose.GTE-mediated induction of CYP3A4 ortholog, the main drug-metabolizing enzyme, could result in decreased efficacy of simultaneously or subsequently administered drug in obese individuals.
Keywords: Green tea extract; Catechins; Drug-metabolizing enzymes; Obesity; Metabolic syndrome

Increased iron bioavailability from lactic-fermented vegetables is likely an effect of promoting the formation of ferric iron (Fe3+) by Nathalie Scheers; Lena Rossander-Hulthen; Inga Torsdottir; Ann-Sofie Sandberg (373-382).
Lactic fermentation of foods increases the availability of iron as shown in a number of studies throughout the years. Several explanations have been provided such as decreased content of inhibitory phytate, increased solubility of iron, and increased content of lactic acid in the fermented product. However, to our knowledge, there are no data to support that the bioavailability of iron is affected by lactic fermentation. The objective of the present study was to investigate whether the bioavailability of iron from a vegetable mix was affected by lactic fermentation and to propose a mechanism for such an event, by conducting human and cell (Caco-2, HepG2) studies and iron speciation measurements (voltammetry). We also investigated whether the absorption of zinc was affected by the lactic fermentation.In human subjects, we observed that lactic-fermented vegetables served with both a high-phytate and low-phytate meal increased the absorption of iron, but not zinc. In vitro digested fermented vegetables were able to provoke a greater hepcidin response per ng Fe than fresh vegetables, indicating that Fe in the fermented mixes was more bioavailable, independent on the soluble Fe content. We measured that hydrated Fe3+ species were increased after the lactic fermentation, while there was no significant change in hydrated Fe2+. Furthermore, lactate addition to Caco-2 cells did not affect ferritin formation in response to Fe nor did lactate affect the hepcidin response in the Caco-2/HepG2 cell system.The mechanism for the increased bioavailability of iron from lactic-fermented vegetables is likely an effect of the increase in ferric iron (Fe3+) species caused by the lactic fermentation. No effect on zinc bioavailability was observed.
Keywords: Lactic fermentation; Iron; Absorption; Ferric; Bioavailability

A dose–response study of vitamin D3 supplementation in healthy Chinese: a 5-arm randomized, placebo-controlled trial by Pang Yao; Ling Lu; Yao Hu; Gang Liu; Xiafei Chen; Liang Sun; Xingwang Ye; He Zheng; Yan Chen; Frank B. Hu; Huaixing Li; Xu Lin (383-392).
To determine the dose–response of vitamin D3 supplementation on serum 25-hydroxyvitamin D [25(OH)D] among Chinese adults.In this 5-arm, randomized, double-blinded controlled trial, 76 healthy participants were assigned to orally administrate 0, 400, 800, 1200 or 2000 IU/d of vitamin D3 for 16 weeks. Serum 25(OH)D, parathyroid hormone, calcium, biomarkers of liver and renal function were measured at multiple time points. The mean (SD) serum 25(OH)D at baseline was 31.6 (8.7) nmol/L, and the dose–response relationship was curvilinear with a plateau around 6 weeks for all doses. At week 16, 25(OH)D was increased by 6.0 (6.5), 21.7 (15.8), 26.3 (12.6), 32.0 (12.8) and 36.3 (26.0) nmol/L for 0, 400, 800, 1200 and 2000 IU/d (all P ≤ 0.002), corresponding to approximately 19, 53, 67, 77 and 80 % of reversion of vitamin D deficiency, respectively. Daily intake of 800 IU vitamin D3 reached a targeted 25(OH)D ≥ 30 nmol/L in at least 97.5 % of Chinese, but not a targeted 25(OH)D ≥ 50 nmol/L even with 2000 IU/d. Change of 25(OH)D was inversely associated with change of PTH concentration (r = −0.39, P < 0.001) after controlling for age and sex. No between-group differences were observed in terms of the change in serum calcium, alanine transaminase, aspartate aminotransferase, gamma-glutamyltransferase and creatinine (P ≥ 0.22).Supplementation with 400, 800, 1200 or 2000 IU/d vitamin D could improve the vitamin D deficiency with various degrees. Whether 2000 IU/d vitamin D3 would generate a better result without side effect requires more studies with larger samples in future.
Keywords: Vitamin D3 supplementation; Serum 25-hydroxyvitamin D; Dose–response; Safety

Low serum 25-hydroxyvitamin D [25(OH)D] levels have been associated with higher risk of many diseases that affect mortality, including cardiovascular disease (CVD) and cancer. The inverse association between serum 25(OH)D and mortality may be modified by excess circulating vitamin A, due to interactions of vitamin A at the level of the vitamin D nuclear receptor. In this prospective cohort study, we investigated whether the association of 25(OH)D with all-cause, cancer, and CVD mortality was modified by circulating vitamin A or preformed vitamin A intake from supplements.We analyzed 15,998 adults in the Third National Health and Nutrition Examination Survey (NHANES III), 1988–1994. Mortality data for all-cause (n = 3890), cancer (n = 844), and CVD mortality (n = 1715) were assessed through December 2006. Serum 25(OH)D was measured using a radioimmunoassay kit, vitamin A biomarkers were measured by HPLC, and information on supplement use was obtained by self-report. Multivariable hazard ratios (HRs) and corresponding 95 % confidence intervals (CI) were estimated by proportional hazards regression.Serum 25(OH)D was significantly inversely associated with all-cause mortality (HR 0.93, 95 % CI 0.89, 0.97, per 10 ng/mL increase) and also with CVD mortality and mortality due to non-cancer/non-cardiovascular causes, but not with cancer mortality. The observed inverse associations remained statistically significant only among participants with serum retinyl esters <7.0 μg/dL. High intake (>5000 IU/day) of preformed vitamin A from supplements attenuated the inverse association of 25(OH)D with overall mortality. The observed interactions were not statistically significant.25(OH)D was inversely associated with overall mortality, CVD mortality, and mortality due to non-cancer/non-CVD causes, but not with cancer mortality. A possible interaction between vitamin A exposure and 25(OH)D concentration appears to be associated with an attenuation of the inverse association between risk of death and quartile of 25(OH)D concentration.
Keywords: Vitamin D; Vitamin A; Mortality; NHANES III

Effect of neonatal malnutrition on expression of nitric oxide synthase enzyme, production of free radicals and in vitro viability of alveolar macrophages infected with methicillin-sensitive and methicillin-resistant Staphylococcus aureus by Natália Gomes de Morais; Thacianna Barreto da Costa; Amanda Lúcia Farias Pedrosa; Maria Carolina Accioly Brelaz de Castro; Suênia Cunha da Gonçalves de Albuquerque; Valéria Rêgo Alves Pereira; Milena de Paiva Cavalcanti; Célia Maria Machado Barbosa de Castro (403-411).
Evaluate the effects of neonatal malnutrition on the microbicidal response and viability of in vitro macrophages infected with Staphylococcus aureus sensitive/resistant to methicillin.Male Wistar rats (n = 24) were divided into two distinct groups: nourished (rats breast-fed by mothers undergoing diet with 17 % casein) and malnourished (rats breast-fed by mothers undergoing diet with 8 % casein). Macrophages were recovered after surgical tracheostomy procedure by collecting bronchoalveolar lavage. Four systems were established: negative control, composed only by phagocytes; positive control, macrophages plus lipopolysaccharide; and two test systems, macrophages plus Staphylococcus aureus sensitive and resistant to methicillin. Plates were incubated at 37 °C for 24 h. After this period, tests for the analysis of cell viability and microbicidal response were performed. In the statistical analysis, the Student’s t and ANOVA tests were used, accepting p < 0.05.The neonatal malnutrition impaired the animals’ body weight. There was a lower expression of the inducible nitric oxide enzyme (iNOS), nitric oxide production, and viability of macrophages infected with methicillin-resistant Staphylococcus aureus. However, increased production of superoxide anion in the malnourished group was detected.Neonatal malnutrition focusing on critical periods of development promoted lower expression of iNOS, nitric oxide production, cell viability, and exacerbated reactive oxygen species production. The high levels of reactive oxygen species may favor the onset of serious and systemic infections with fatal outcome if associated with methicillin-resistant Staphylococcus aureus.
Keywords: Index terms: malnutrition; Macrophages; Staphylococcus aureus ; Methicillin

Comparing the metabolism of quercetin in rats, mice and gerbils by Shu-Lan Yeh; Yi-Chin Lin; Yi-Ling Lin; Chien-Chun Li; Cheng-Hung Chuang (413-422).
Several species of rodents are used to investigate the metabolism of quercetin in vivo. However, it is unclear whether they are a proper animal model. Thus, we compared the metabolism of quercetin in Wistar rats (rats), Balb/c mice (mice) and Mongolian gerbils (gerbils).We determined the levels of quercetin metabolites, quercetin-3-glucuronide (Q3G), quercetin-3′-sulfate (Q3′S) and methyl-quercetin isorhamnetin (IH), in the plasma, lungs and livers of three species of animals by high-performance liquid chromatography after acute and/or chronic quercetin administration. The metabolic enzyme activities in the intestinal mucosal membrane and liver were also investigated.First, we found that after acute quercetin administration, the Q3′S level was the highest in gerbils. However, after long-term supplementation (20 weeks), Q3G was the dominant metabolite in the plasma, lungs and livers followed by IH and Q3′S in all animals, although the gerbils still had a higher Q3′S conversion ratio. The average concentrations of total quercetin concentration in the plasma of gerbils were the highest in both short- and long-term studies. The activities of uridine 5′-diphosphate-glucuronosyltransferase, phenolsulfotransferase and catechol-O-methyltransferase were induced by quercetin in a dose- and tissue-dependent manner in all animals.Taken together, in general, after long-term supplementation the metabolism of quercetin is similar in all animals and is comparable to that of humans. However, the accumulation of quercetin and Q3′S conversion ratio in gerbils are higher than those in the other animals.
Keywords: Quercetin-3-glucuronide; Quercetin-3′-sulfate; Uridine 5′-diphosphate- glucuronosyltransferase; Phenolsulfotransferase; Catechol-O-methyltransferase