European Journal of Nutrition (v.57, #2)

Healthy ageing: the natural consequences of good nutrition—a conference report by D. Marsman; D. W. Belsky; D. Gregori; M. A. Johnson; T. Low Dog; S. Meydani; S. Pigat; R. Sadana; A. Shao; J. C. Griffiths (15-34).
Many countries are witnessing a marked increase in longevity and with this increased lifespan and the desire for healthy ageing, many, however, suffer from the opposite including mental and physical deterioration, lost productivity and quality of life, and increased medical costs. While adequate nutrition is fundamental for good health, it remains unclear what impact various dietary interventions may have on prolonging good quality of life. Studies which span age, geography and income all suggest that access to quality foods, host immunity and response to inflammation/infections, impaired senses (i.e., sight, taste, smell) or mobility are all factors which can limit intake or increase the body’s need for specific micronutrients. New clinical studies of healthy ageing are needed and quantitative biomarkers are an essential component, particularly tools which can measure improvements in physiological integrity throughout life, thought to be a primary contributor to a long and productive life (a healthy “lifespan”). A framework for progress has recently been proposed in a WHO report which takes a broad, person-centered focus on healthy ageing, emphasizing the need to better understand an individual’s intrinsic capacity, their functional abilities at various life stages, and the impact by mental, and physical health, and the environments they inhabit.
Keywords: Ageing; Biomarkers; Centenarians; Functional ability; Geroprotectors; Immunosenescence; Intrinsic capacity; Lifespan; Micronutrients; Minerals; Nutrition; Quality of life; Vitamins

Does soy protein affect circulating levels of unbound IGF-1? by Mark Messina; Pamela Magee (423-432).
Despite the enormous amount of research that has been conducted on the role of soyfoods in the prevention and treatment of chronic disease, the mechanisms by which soy exerts its physiological effects are not fully understood. The clinical data show that neither soyfoods nor soy protein nor isoflavones affect circulating levels of reproductive hormones in men or women. However, some research suggests that soy protein, but not isoflavones, affects insulin-like growth factor I (IGF-1).Since IGF-1 may have wide-ranging physiological effects, we sought to determine the effect of soy protein on IGF-1 and its major binding protein insulin-like growth factor-binding protein (IGFBP-3). Six clinical studies were identified that compared soy protein with a control protein, albeit only two studies measured IGFBP-3 in addition to IGF-1.Although the data are difficult to interpret because of the different experimental designs employed, there is some evidence that large amounts of soy protein (>25 g/day) modestly increase IGF-1 levels above levels observed with the control protein.The clinical data suggest that a decision to incorporate soy into the diet should not be based on its possible effects on IGF-1.
Keywords: Soy; Isoflavones; IGF-1; IGFBP-3; Cancer; Clinical trials

Gluten- and casein-free diet and autism spectrum disorders in children: a systematic review by Anna Piwowarczyk; Andrea Horvath; Jan Łukasik; Ewa Pisula; Hania Szajewska (433-440).
Effective treatments for core symptoms of autism spectrum disorders (ASD) are lacking. We systematically updated evidence on the effectiveness of a gluten-free and casein-free (GFCF) diet as a treatment for ASD in children.The Cochrane Library, MEDLINE, and EMBASE databases were searched up until August 2016, for randomized controlled trials (RCTs); additional references were obtained from reviewed articles.Six RCTs (214 participants) were included. With few exceptions, there were no statistically significant differences in autism spectrum disorder core symptoms between groups, as measured by standardized scales. One trial found that compared with the control group, in the GFCF diet group there were significant improvements in the scores for the ‘communication’ subdomain of the Autism Diagnostic Observation Schedule and for the ‘social interaction’ subdomain of the Gilliam Autism Rating Scale. Another trial found significant differences between groups in the post-intervention scores for the ‘autistic traits’, ‘communication’, and ‘social contact’ subdomains of a standardized Danish scheme. The remaining differences, if present, referred to parent-based assessment tools or other developmental/ASD-related features. No adverse events associated with a GFCF diet were reported.Overall, there is little evidence that a GFCF diet is beneficial for the symptoms of ASD in children.
Keywords: Randomized controlled trial; Autism spectrum disorders; Children; Gluten; Casein

Soy isoflavone intake and prevalence of depressive symptoms during pregnancy in Japan: baseline data from the Kyushu Okinawa Maternal and Child Health Study by Yoshihiro Miyake; Keiko Tanaka; Hitomi Okubo; Satoshi Sasaki; Shinya Furukawa; Masashi Arakawa (441-450).
Several observational studies and trials examined the relationship between isoflavones or soybeans and depressive symptoms among peri- and postmenopausal women. We cross-sectionally evaluated the associations between intake of soy products and isoflavones and depressive symptoms during pregnancy in Japan.Study subjects were 1745 pregnant women. Dietary intake during the preceding month was assessed using a self-administered diet history questionnaire. Depressive symptoms were defined by a score of 16 or over in the Center for Epidemiologic Studies Depression Scale.Higher intake of total soy products, tofu, tofu products, fermented soybeans, boiled soybeans, miso soup, and isoflavones was independently related to a lower prevalence of depressive symptoms during pregnancy: The adjusted prevalence ratios (95 % confidence intervals, P for trend) between extreme quartiles were 0.63 (0.47–0.85, 0.002), 0.72 (0.54–0.96, 0.007), 0.74 (0.56–0.98, 0.04), 0.57 (0.42–0.76, <0.0001), 0.73 (0.55–0.98, 0.03), 0.65 (0.49–0.87, 0.003), and 0.63 (0.46–0.86, 0.002), respectively. A significant positive exposure–response relationship was found between miso intake and depressive symptoms during pregnancy. No material relationship was observed between soymilk intake and depressive symptoms during pregnancy.Our study is the first to show independent inverse relationships between intake of total soy products, tofu, tofu products, fermented soybeans, boiled soybeans, miso soup, and isoflavones and depressive symptoms during pregnancy.
Keywords: Depressive symptoms; Intake; Isoflavones; Japanese; Pregnancy; Soy

Dietary intake and biomarker status of folate in Swedish adults by Veronica Öhrvik; Eva Warensjö Lemming; Cecilia Nälsén; Wulf Becker; Peter Ridefelt; Anna Karin Lindroos (451-462).
National data on folate status are missing in Sweden, and regional data indicate folate insufficiency in up to more than 25% of the study populations. The objectives were to determine folate intake and status in the adult Swedish population as well as identifying dietary patterns associated with beneficial folate status.Folate intake was estimated using a web-based 4-d food record in adults aged 18–80 years (n = 1797). Folate status was measured as erythrocyte (n = 282) and plasma folate concentrations (n = 294). Factor analysis was used to derive a dietary pattern associated with a higher folate status.Median folate intake was 246 µg/day (Q 1 = 196, Q 3 = 304, n = 1797) and for women of reproductive age 227 µg/day (Q 1 = 181, Q 3 = 282, n = 450). As dietary folate equivalents (DFE), median intake was 257 µg/day (Q 1 = 201, Q 3 = 323) and for women of reproductive age 239 µg/day (Q 1 = 185, Q 3 = 300). Low blood folate concentrations were found in 2% (erythrocyte concentrations <317 nmol/L) and 4% (plasma concentrations <6.8 nmol/L) of the participants, respectively. None of the women of reproductive age had erythrocyte folate concentrations associated with the lowest risk of neural tube defects. Dietary patterns associated with higher folate status were rich in vegetables, pulses and roots as well as cheese and alcoholic beverages, and low in meat.Prevalence of low erythrocyte folate concentrations was low in this population, and estimated dietary intakes are well above average requirement. However, to obtain a folate status optimal for prevention of neural tube defects major dietary changes are required and folic acid supplements recommended prior to conception.
Keywords: Folate intake; Folate status; Red blood cell folate concentrations; Riksmaten adults 2010–11; Food intake; Swedish national dietary survey

A mixture of milk and vegetable lipids in infant formula changes gut digestion, mucosal immunity and microbiota composition in neonatal piglets by Isabelle Le Huërou-Luron; Karima Bouzerzour; Stéphanie Ferret-Bernard; Olivia Ménard; Laurence Le Normand; Cécile Perrier; Cindy Le Bourgot; Julien Jardin; Claire Bourlieu; Thomas Carton; Pascale Le Ruyet; Isabelle Cuinet; Cécile Bonhomme; Didier Dupont (463-476).
Although composition of infant formula has been significantly improved during the last decade, major differences with the composition and structure of breast milk still remain and might affect nutrient digestion and gut biology. We hypothesized that the incorporation of dairy fat in infant formulas could modify their physiological impacts by making their composition closer to that of human milk. The effect of milk fat and milk fat globule membrane (MFGM) fragments in infant formulas on gut digestion, mucosal immunity and microbiota composition was evaluated.Three formulas containing either (1) vegetable lipids stabilized only by proteins (V-P), (2) vegetable lipids stabilized by a mixture of proteins and MFGM fragments (V-M) and (3) a mixture of milk and vegetable lipids stabilized by a mixture of proteins and MFGM fragments (M-M) were automatically distributed to 42 newborn piglets until slaughter at postnatal day (PND) 7 or 28, and compared to a fourth group of sow’s suckling piglets (SM) used as a breast-fed reference.At both PND, casein and β-lactoglobulin digestion was reduced in M-M proximal jejunum and ileum contents compared to V-P and V-M ones leading to more numerous β-Cn peptides in M-M contents. The IFNγ cytokine secretion of ConA-stimulated MLN cells from M-M piglets tended to be higher than in V-P ones at PND 7 and PND 28 and was closer to that of SM piglets. No dietary treatment effect was observed on IL-10 MLN cell secretion. Changes in faecal microbiota in M-M piglets resulted in an increase in Proteobacteria and Bacteroidetes and a decrease in Firmicutes phyla compared to V-P ones. M-M piglets showed higher abundances of Parabacteroides, Escherichia/Shigella and Klebsiella genus.The incorporation of both milk fat and MFGM fragments in infant formula modifies protein digestion, the dynamic of the immune system maturation and the faecal microbiota composition.
Keywords: Infant formula; Milk fat; Digestion; Intestinal permeability; Intestinal immune system; Microbiota

Methylglyoxal treatment in lactating mothers leads to type 2 diabetes phenotype in male rat offspring at adulthood by Flávio Andrade Francisco; Luiz Felipe Barella; Sandra da Silva Silveira; Lucas Paulo Jacinto Saavedra; Kelly Valério Prates; Vander Silva Alves; Claudinéia Conationi da Silva Franco; Rosiane Aparecida Miranda; Tatiane Aparecida Ribeiro; Laize Peron Tófolo; Ananda Malta; Elaine Vieira; Kesia Palma-Rigo; Audrei Pavanello; Isabela Peixoto Martins; Veridiana Mota Moreira; Júlio Cezar de Oliveira; Paulo Cezar de Freitas Mathias; Rodrigo Mello Gomes (477-486).
Environmental and nutritional disorders during perinatal period cause metabolic dysfunction in the progeny and impair human health. Advanced glycation end products (AGEs) are primarily produced during metabolism of excess blood glucose, which is observed in diabetes. Methylglyoxal (MG) is a precursor for the generation of endogenous AGEs, which disturbs the metabolism. This work aimed to investigate whether the maternal MG treatment during lactation programs the progeny to metabolic dysfunction later in life.Female Wistar rats were divided into two groups: control group (C) treated with saline and MG group treated with MG (60 mg/kg/day) by gavage throughout the lactation period. Both mothers and offspring were fed a standard chow. At weaning, breast milk composition was analyzed and mothers euthanized for blood and tissue sample collections. At 90 days of age, offspring were submitted to glucose tolerance test (ivGTT) and euthanized for blood and tissue samples collection.MG mothers showed increase in glucose and fructosamine levels; however, they showed low insulin levels and failure in β-cell function (p < 0.05). MG mothers also showed dyslipidemia (p < 0.05). Moreover, breast milk had elevated levels of glucose, triglycerides, cholesterol and fructosamine and low insulin (p < 0.05). Interestingly, MG offspring had increased body weight and adipose tissue at adulthood, and they also showed glucose intolerance and failure in β-cell function (p < 0.05). Besides, MG offspring showed dyslipidemia (p < 0.05) increasing cardiovascular diseases risk.Maternal MG treatment negatively affects the male rat offspring, leading to type 2 diabetes and dyslipidemia in later life, possibly by changes in breast milk composition.
Keywords: Diabetes; AGEs; Methylglyoxal; Dyslipidemia; Lactation; Metabolic programming

Bioactive compounds from regular diet and faecal microbial metabolites by Tania Fernández-Navarro; Nuria Salazar; Isabel Gutiérrez-Díaz; Borja Sánchez; Patricia Rúas-Madiedo; Clara G. de los Reyes-Gavilán; Abelardo Margolles; Miguel Gueimonde; Sonia González (487-497).
Short-chain fatty acids (SCFAs) formation by intestinal bacteria is regulated by many different factors, among which dietary fibre is currently receiving most attention. However, since fibre-rich foods are usually good dietary sources of phenolic compounds, which are also known to affect the microbiota, authors hypothesize that the regular intake of these bioactive compounds could be associated with a modulation of faecal SCFA production by the intestinal microbiota.In this work, food intake was recorded by means of a validated Food Frequency Questionnaire. Fibres were determined using Marlett food composition tables, and phenolic compounds were obtained from Phenol-Explorer Database. Analysis of SCFA was performed by gas chromatography–flame ionization/mass spectrometry and quantification of microbial populations in faeces by quantitative PCR.Klason lignin and its food contributors, as predictors of faecal butyrate production, were directly associated with Bacteroides and Bifidobacterium levels, as well as lignans with Bacteroides. Also, anthocyanidins, provided by strawberries, were associated with faecal propionate and inversely related to Lactobacillus group.These results support the hypothesis we put forward regarding the association between some vegetable foods (strawberries, pasta, lentils, lettuce and olive oil) and faecal SCFA. More studies are needed in order to elucidate whether these associations have been mediated by the bacterial modulatory effect of the bioactive compounds, anthocyanins, lignans or Klason lignin, present in foodstuffs.
Keywords: Fibre; Polyphenol; Microbiota; Short-chain fatty acids

Effect of soy on metabolic syndrome and cardiovascular risk factors: a randomized controlled trial by Massimiliano Ruscica; Chiara Pavanello; Sara Gandini; Monica Gomaraschi; Cecilia Vitali; Chiara Macchi; Beatrice Morlotti; Gilda Aiello; Raffaella Bosisio; Laura Calabresi; Anna Arnoldi; Cesare R. Sirtori; Paolo Magni (499-511).
Cardiovascular diseases are currently the commonest cause of death worldwide. Different strategies for their primary prevention have been planned, taking into account the main known risk factors, which include an atherogenic lipid profile and visceral fat excess.The study was designed as a randomized, parallel, single-center study with a nutritional intervention duration of 12 weeks. Whole soy foods corresponding to 30 g/day soy protein were given in substitution of animal foods containing the same protein amount.Soy nutritional intervention resulted in a reduction in the number of MetS features in 13/26 subjects. Moreover, in the soy group we observed a significant improvement of median percentage changes for body weight (−1.5 %) and BMI (−1.5 %), as well as for atherogenic lipid markers, namely TC (−4.85 %), LDL-C (−5.25 %), non-HDL-C (−7.14 %) and apoB (−14.8 %). Since the majority of the studied variables were strongly correlated, three factors were identified which explained the majority (52 %) of the total variance in the whole data set. Among them, factor 1, which loaded lipid and adipose variables, explained the 22 % of total variance, showing a statistically significant difference between treatment arms (p = 0.002).The inclusion of whole soy foods (corresponding to 30 g/day protein) in a lipid-lowering diet significantly improved a relevant set of biomarkers associated with cardiovascular risk.
Keywords: Soy protein; Lipids; Metabolic syndrome and obesity

Differences in food intake and genetic variability in taste receptors between Czech pregnant women with and without gestational diabetes mellitus by Vendula Bartáková; Katarína Kuricová; Filip Zlámal; Jana Bělobrádková; Katetřina Kaňková (513-521).
Gestational diabetes mellitus (GDM) represents the most frequent metabolic disorder in pregnancy. Since dietary intake plays an important role in obesity and type 2 diabetes development, it is likely to be for the susceptibility to GDM too. Food preferences, driving partly the diet composition, are changing during pregnancy. Taste and genetic variability in taste receptors is an important factor in determining food preferences. Aims of our study were (1) to characterize dietary habits of pregnant women and to find possible differences in food preferences between healthy pregnant women and those with GDM and (2) to ascertain possible association of several single nucleotide polymorphisms (SNPs) in taste receptor (TR) genes with GDM.A total of 363 pregnant women (293 with GDM and 70 with physiologic pregnancy) were included in the study. Dietary pattern spanning the period of approx. 6 months preceding the time of GDM screening was assessed using a semi-quantitative food frequency questionnaire. A total of five SNPs in TR genes were selected for genotyping based on their functionality or previous associations.Women with GDM exhibited significantly more frequent meat consumption (esp. poultry, pork and smoked meat), dairy products and sweet beverages consumption. The legumes consumption was found to be inversely correlated with fasting glycaemia (P = 0.007, Spearman). CC genotype in TAS2R9 gene (SNP rs3741845) was significantly associated with GDM (P = 0.0087, Chi-square test).Our study showed differences in dietary intake of selected food items between healthy pregnant women and those with GDM and genetic association of bitter taste receptor allele with GDM.
Keywords: Gestational diabetes mellitus; Food preferences; Dietary intake; Taste receptors; Single nucleotide polymorphism; Genetic variability

Considering the low content in breast milk breastfed infants might be at particular risk for depleted iron stores after the first months of life. This study evaluates the association of the mode of milk feeding and the timing of complementary food (CF) introduction with parameters of iron status in term healthy infants in Germany.In this secondary analysis of a randomized controlled trial, parents recorded all foods consumed by their infants from the age of 8 weeks onwards. Mothers were advised on the German food-based dietary guidelines for infants. Accordingly, CF was introduced between the fifth and seventh month of age. Blood samples were taken at 4 and at 10 months of age for analyses of iron status parameters. Iron depletion was defined as serum ferritin <12 ng/mL.The iron intake was lower in breastfed infants (n = 50) than in formula fed (n = 23) with decreasing differences during the course of infancy. At 10 months of age, most iron parameters were not associated with the mode of milk feeding or the timing of CF introduction. At this age, the iron depletion prevalence was >34% without general differences according to the mode of milk feeding or the timing of CF introduction.The high prevalence of depleted iron stores observed in both breastfed and formula-fed infants illustrates the need for further studies to improve our understanding of the optimal iron intake and sensitive parameters of iron status in infancy.
Keywords: Breastfeeding; Formula; Complementary feeding; Iron status; Iron deficiency

Coffee and oxidative stress: a human intervention study by Sergey Shaposhnikov; Thomas Hatzold; Naouale El Yamani; Philip Mark Stavro; Yolanda Lorenzo; Maria Dusinska; Astrid Reus; Wilrike Pasman; Andrew Collins (533-544).
Coffee is known to contain phytochemicals with antioxidant potential. The aim of this study was to investigate possible antioxidant effects of coffee in healthy human volunteers.A placebo-controlled intervention trial was carried out on 160 healthy human subjects, randomised into three groups, receiving 3 or 5 cups of study coffee or water per day, for 8 weeks. Blood samples were taken before, during, and after the intervention. Serum was used for analysis of blood lipids and standard clinical chemistry analytes. Peripheral blood mononuclear cells were isolated, and DNA damage (strand breaks and oxidised bases) was measured with the comet assay. The lipid oxidation product isoprostane 8-iso-PGF2α was assayed in urine samples by LC–MS/MS.There was no significant effect of coffee consumption on the markers of oxidation of DNA and lipids. Creatinine (in serum) increased by a few per cent in all groups, and the liver enzyme γ-glutamyl transaminase was significantly elevated in serum in the 5 cups/day group. Other clinical markers (including glucose and insulin), cholesterol, triacylglycerides, and inflammatory markers were unchanged. There was no effect of coffee on blood pressure.In a carefully controlled clinical trial with healthy subjects, up to 5 cups of coffee per day had no detectable effect, either beneficial or harmful, on human health.
Keywords: Human intervention trial; Coffee; DNA oxidation; Lipid oxidation

Associations between exclusive breastfeeding and physical fitness during childhood by Mahmoud Zaqout; Nathalie Michels; Wolfgang Ahrens; Claudia Börnhorst; Dénes Molnár; Luis A. Moreno; Gabriele Eiben; Alfonso Siani; Stalo Papoutsou; Toomas Veidebaum; Stefaan De Henauw (545-555).
Exposure to breastfeeding improves the survival, health, and development of children; therefore, breast milk is recommended as the exclusive nutrient source for feeding term infants during the first 6 months. This cross-sectional study aimed to determine the possible association between exposure to exclusive breastfeeding and physical fitness performance in children and, if so, whether this association is influenced by the breastfeeding duration.A total of 2853 (52.3 % girls) European children from the IDEFICS study aged 6–11 years with complete data on physical fitness (cardiorespiratory fitness, muscular strength, flexibility, balance, speed) and exclusive breastfeeding duration (never, 1–3, 4–6, 7–12 months) were included in the present study. Multivariate and mixed linear regression models were estimated and adjusted for sex, age, birth weight, diet, physical activity, body mass index, and parental factors (age, body mass index, educational attainment).We found a positive association between exclusive breastfeeding and lower-body explosive strength (β = 0.034) as well as flexibility (β = 0.028). We also found a positive association between breastfeeding and balance in boys (β = 0.039), while this association was negative in girls (β = −0.029). To improve lower-body explosive strength, 1–3 months of exclusive breastfeeding were enough; a longer duration did not lead to increasing benefit. In contrast, 4–6 months of breastfeeding were necessary to have any benefit on flexibility or balance, although this became nonsignificant after adjustment for body mass index and physical activity.Exclusive breastfeeding seems a natural way of slightly improving some physical fitness components (mainly lower-body muscle strength) and thus future health.
Keywords: Exclusive breastfeeding; Physical fitness; Children; Muscle strength; Flexibility; Balance

l-Arginine and B vitamins improve endothelial function in subjects with mild to moderate blood pressure elevation by Daniel Menzel; Hermann Haller; Manfred Wilhelm; Horst Robenek (557-568).
The aim of this trial was to investigate the influence of a dietetic product consisting of a unique combination of l-arginine with the vitamins B6, folic acid and B12 (Telcor® Arginin plus) on endothelial dysfunction.Subjects aged 40–65 years with mild to moderate blood pressure (BP) elevation not treated with anti-hypertensive drugs were randomly assigned to either the dietetic product (n = 40) or a matching placebo (n = 41) for 3 months with open follow-up for a further 3 months. Postprandial change in endothelial function was assessed using the validated reactive hyperaemia index (RHI) at 3 months compared to the study onset (RHI post–pre, visit 3–visit 1; ΔΔRHI). Secondary parameters included BP and plasma homocysteine concentration.The primary efficacy analysis revealed superiority of the nutritional intervention over placebo (p = 0.0349) in reducing the deterioration of endothelial function. While in the active group ΔΔRHI increased (0.371 ± 0.122), almost no change could be detected in the placebo group (0.031 ± 0.100), thus demonstrating a significant improvement in vascular function in the intervention group. Moreover, the intervention reduced BP and homocysteine levels. Non-serious adverse events were equally distributed in both groups, and none of the events were assessed as possibly intervention-related by the investigators.This trial confirmed the effective and safe use of dietary management with l-arginine in combination with B vitamins. The primary efficacy analysis demonstrated a statistically significant superiority of the combination of l-arginine with B vitamins over placebo in improving and restoring impaired endothelial function and lowering BP in patients with mild to moderate blood pressure elevation.
Keywords: Endothelial function; Atherosclerosis; Blood pressure; l-Arginine; B vitamins

Cardiac changes in apoptosis, inflammation, oxidative stress, and nitric oxide system induced by prenatal and postnatal zinc deficiency in male and female rats by Lorena Vanesa Juriol; María Natalia Gobetto; Facundo Mendes Garrido Abregú; Marina Ercilia Dasso; Gonzalo Pineda; Leandro Güttlein; Andrea Carranza; Osvaldo Podhajcer; Jorge Eduardo Toblli; Rosana Elesgaray; Cristina Teresa Arranz; Analía Lorena Tomat (569-583).
Zinc restriction during fetal and postnatal development could program cardiovascular diseases in adulthood. The aim of this study was to determine the effects of zinc restriction during fetal life, lactation, and/or post-weaning growth on cardiac inflammation, apoptosis, oxidative stress, and nitric oxide system of male and female adult rats.Wistar rats were fed a low- or a control zinc diet during pregnancy and up to weaning. Afterward, offspring were fed either a low- or a control zinc diet until 81 days of life. IL-6 and TNF-α levels, TUNEL assay, TGF-β1 expression, thiobarbituric acid-reactive substances that determine lipoperoxidation damage, NADPH oxidase-dependent superoxide anion production, antioxidant and nitric oxide synthase activity, mRNA and protein expression of endothelial nitric oxide synthase, and serine1177 phosphorylation isoform were determined in left ventricle.Zinc deficiency activated apoptotic and inflammatory processes and decreased TGF-β1 expression and nitric oxide synthase activity in cardiac tissue of both sexes. Male zinc-deficient rats showed no changes in endothelial nitric oxide synthase expression, but a lower serine1177 phosphorylation. Zinc deficiency induced an increase in antioxidant enzymes activity and no differences in lipoperoxidation products levels in males. Females were less sensitive to this deficiency exhibiting lower increase in apoptosis, lower decrease in expression of TGF-β1, and higher antioxidant and nitric oxide enzymes activities. A zinc-adequate diet during postnatal life reversed most of these mechanisms.Prenatal and postnatal zinc deficiency induces alterations in cardiac apoptotic, inflammatory, oxidative, and nitric oxide pathways that could predispose the onset of cardiovascular diseases in adult life.
Keywords: Moderate zinc deficiency; Prenatal and postnatal growth; Cardiac apoptosis; Cardiac oxidative stress; Cardiac nitric oxide system; Sex differences

Whey protein consumption after resistance exercise reduces energy intake at a post-exercise meal by Alistair Monteyne; Alex Martin; Liam Jackson; Nick Corrigan; Ellen Stringer; Jack Newey; Penny L. S. Rumbold; Emma J. Stevenson; Lewis J. James (585-592).
Protein consumption after resistance exercise potentiates muscle protein synthesis, but its effects on subsequent appetite in this context are unknown. This study examined appetite and energy intake following consumption of protein- and carbohydrate-containing drinks after resistance exercise.After familiarisation, 15 resistance training males (age 21 ± 1 years, body mass 78.0 ± 11.9 kg, stature 1.78 ± 0.07 m) completed two randomised, double-blind trials, consisting of lower-body resistance exercise, followed by consumption of a whey protein (PRO 23.9 ± 3.6 g protein) or dextrose (CHO 26.5 ± 3.8 g carbohydrate) drink in the 5 min post-exercise. An ad libitum meal was served 60 min later, with subjective appetite measured throughout. Drinks were flavoured and matched for energy content and volume. The PRO drink provided 0.3 g/kg body mass protein.Ad libitum energy intake (PRO 3742 ± 994 kJ; CHO 4172 ± 1132 kJ; P = 0.007) and mean eating rate (PRO 339 ± 102 kJ/min; CHO 405 ± 154 kJ/min; P = 0.009) were lower during PRO. The change in eating rate was associated with the change in energy intake (R = 0.661, P = 0.007). No interaction effects were observed for subjective measures of appetite. The PRO drink was perceived as creamier and thicker, and less pleasant, sweet and refreshing (P < 0.05).These results suggest whey protein consumption after resistance exercise reduces subsequent energy intake, and this might be partially mediated by a reduced eating rate. Whilst this reduced energy intake is unlikely to impair hypertrophy, it may be of value in supporting an energy deficit for weight loss.
Keywords: Appetite; Energy balance; Weight management; Protein synthesis; Anabolism; Body composition

Effect of buffalo casein-derived novel bioactive peptides on osteoblast differentiation by Srinu Reddi; Venkatesa Perumal Shanmugam; Kemgang Sonfack Tanedjeu; Suman Kapila; Rajeev Kapila (593-605).
Epidemiological and intervention studies show that milk consumption in childhood and during adolescence is related to higher bone mineral density. Milk and milk products prevent the bone loss in pre- and postmenopausal women. Apart from calcium, there are other biologically active compounds in milk such as bioactive peptides which may play a role in promoting bone health. Casein is the major protein in milk which has also been reported to have numerous biological active peptides within it. The hypothesis of the present study was to identify the key peptides behind osteoanabolic nature of the milk protein, which further can be used to prepare functional foods to alleviate bone diseases like osteoporosis. Hence, this study was carried out to investigate osteogenic nature of four novel bioactive peptides [PEP1 (EDVPSER), PEP2 (NAVPITPTL), PEP3 (VLPVPQK) and PEP4 (HPHPHLSF)] derived from buffalo casein by in vitro osteoblast differentiation model.Calvaria cells were isolated from 3-day-old rat pups, cultured under in vitro conditions till confluence and further used for experiments. Calvarial osteoblast cells were cultured in the presence or absence of peptides including positive controls up to 21 days. Effect of peptides was checked at regular intervals by quantifying osteoblast differentiation marker genes (ALP, OCN and COL-1) expression, alkaline phosphatase activity, osteocalcin level in culture supernatants, mineral deposition by alizarin red staining and caspase-3 and 9 assays.The osteoblast differentiation marker genes (ALP, OCN and COL-1) expression was significantly [(p < 0.01) (p < 0.001)] up-regulated in the presence of these peptides. The peptides also significantly induced alkaline phosphatase activity, osteocalcin level and mineral deposition in comparison with the control. It was also observed that all the four peptides did not show any cytotoxic effect during 21-day treatment period.All peptides enhanced osteoblast differentiation along with the positive controls. These results hold an immense scope to use peptides as preventive measure for reducing incidence of osteoporosis. These peptides can also be used as drugs and can be utilized as functional ingredients in functional foods preparation for osteoporosis therapy, but in vivo studies are required for further confirmation.
Keywords: Bioactive peptides; Milk; Casein; Osteoblast; Osteoporosis

Short-term effects of a green coffee extract-, Garcinia c ambogia- and l-carnitine-containing chewing gum on snack intake and appetite regulation by Cecilia Bobillo; Graham Finlayson; Ana Martínez; Daniela Fischman; Analisa Beneitez; Alejandro J. Ferrero; Belisario E. Fernández; Marcos A. Mayer (607-615).
Different studies have assessed the influence of chewing gum to aid control of appetite and reduce food intake.The aims of the present study were to evaluate the effects of chewing gum on satiety, food hedonics and snack intake and to explore the potential effects of the combination of Garcinia c ambogia, green coffee extract and l-carnitine on satiety, when administered in a gum format.This was a prospective study in which 57 subjects randomly received three kinds of treatments, in a crossover design: (1) active gum; (2) placebo gum; and (3) no gum. Food preferences and appetite sensations were evaluated by means of the Leeds Food Preference Questionnaire and visual analog scales.There was a significant reduction in low-fat sweet snack intake with placebo gum and the active gum compared to no gum and a reduction in high-fat sweet snack intake with the active gum compared to placebo gum and no gum. Total caloric intake was only reduced in the active gum condition. Both the active and placebo gum conditions significantly reduced hunger and prospective food consumption and increased fullness compared to no gum and were associated with a reduced wanting for sweet food in the LFPQ, consistent in a reduction in the relative preference for sweet snacks versus savoury snacks.This study supports the notion that chewing gum containing nutraceutical products might aid in the control over snack intake and reduce hunger sensations.
Keywords: Green coffee; Garcinia c ambogia ; l-Carnitine; Chewing gum; Satiety

Estimated dietary intake and major food sources of polyphenols in elderly of Viçosa, Brazil: a population-based study by Mary Anne Nascimento-Souza; Pedro Gontijo de Paiva; Jara Pérez-Jiménez; Sylvia do Carmo Castro Franceschini; Andréia Queiroz Ribeiro (617-627).
Epidemiological studies have remarked the beneficial role that polyphenols may have in the elderly population such as cancer and cardiovascular disease prevention. This is particularly relevant considering the global tendency of population aging. Data on polyphenol intake in the elderly population are scarce and usually provide partial information—only for some polyphenol classes. The aim of this study was to estimate the intake of polyphenols and its major dietary contributors in the population of Viçosa.A cross-sectional population-based survey including 620 elderly was conducted in Viçosa, Brazil. Food intake was estimated by recall of habitual consumption (RHC). Polyphenol intake was calculated by matching food consumption data from the RHC with the polyphenol content in foods listed in the Phenol-Explorer database.The average total polyphenol intake was 1198.6 mg/day (533.7 mg/day as aglycone). The main polyphenol classes were phenolic acids (729.5 mg/day) and flavonoids (444.7 mg/day). The main dietary contributors for total polyphenols were coffee (45.8%), beans (32.8%) and polenta (1.3%). A total of 292 polyphenols divided in 14 classes and 23 subclasses were found. The individual compounds with the highest intake were isomers of chlorogenic acid (i.e., 5-caffeoylquinic acid, 4-caffeoylquinic acid and 3-caffeoylquinic acid) among hydroxycinnamic acids that largely originated from coffee.The data reported here can be used to evaluate the association between the amount and type of ingested polyphenols and health outcomes in epidemiological studies in order to eventually establish nutritional recommendations.
Keywords: Dietary polyphenols; Flavonoids; Phenolic acids; Stilbenes; Lignans; Elderly; Dietary intake

Dietary sources of sugars in adolescents’ diet: the HELENA study by M. I. Mesana; A. Hilbig; O. Androutsos; M. Cuenca-García; J. Dallongeville; I. Huybrechts; S. De Henauw; K. Widhalm; A. Kafatos; E. Nova; A. Marcos; M. González-Gross; D. Molnar; F. Gottrand; L. A. Moreno (629-641).
To report dietary sugars consumption and their different types and food sources, in European adolescents.Food consumption data of selected groups were obtained from 1630 adolescents (45.6% males, 12.5–17.5 years) from the HELENA study using two nonconsecutive 24-h recalls. Energy intake, total sugars and free sugars were assessed using the HELENA-DIAT software. Multiple regression analyses were performed adjusting for relevant confounders.Total sugars intake (137.5 g/day) represented 23.6% and free sugars (110.1 g/day), 19% of energy intake. Girls had significantly lower intakes of energy, carbohydrates, total sugars and free sugars. 94% of adolescents had a consumption of free sugars above 10% of total energy intake. The main food contributor to free sugars was ‘carbonated, soft and isotonic drinks,’ followed by ‘non-chocolate confectionary’ and ‘sugar, honey, jam and syrup.’ Older boys and girls had significantly higher intakes of free sugars from ‘cakes, pies and biscuits.’ Free sugars intake was negatively associated with low socioeconomic status for ‘non-chocolate confectionary’ and ‘sugar, honey and jam’ groups; with low maternal educational level for carbonated and ‘soft drinks,’ ‘sugar, honey and jam,’ ‘cakes and pies’ and ‘breakfast cereals’ groups; and with high paternal educational level for ‘carbonated and soft drinks’ and ‘chocolates’ group.The majority (94%) of studied adolescents consumed free sugars above 10% of daily energy intake. Our data indicate a broad variety in foods providing free sugars. Continued efforts are required at different levels to reduce the intake of free sugars, especially in families with a low educational level.
Keywords: Adolescents; Free sugars; Food sources; Europe; Diet

Meat intake, cooking methods and doneness and risk of colorectal tumours in the Spanish multicase-control study (MCC-Spain) by Jordi de Batlle; Esther Gracia-Lavedan; Dora Romaguera; Michelle Mendez; Gemma Castaño-Vinyals; Vicente Martín; Núria Aragonés; Inés Gómez-Acebo; Rocío Olmedo-Requena; José Juan Jimenez-Moleon; Marcela Guevara; Mikel Azpiri; Cristóbal Llorens-Ivorra; Guillermo Fernandez-Tardon; Jose Andrés Lorca; José María Huerta; Victor Moreno; Elena Boldo; Beatriz Pérez-Gómez; Jesús Castilla; Tania Fernández-Villa; Juan Pablo Barrio; Montserrat Andreu; Antoni Castells; Trinidad Dierssen; Jone M. Altzibar; Manolis Kogevinas; Marina Pollán; Pilar Amiano (643-653).
Although there is convincing evidence that red and processed meat intake increases the risk of colorectal cancer (CRC), the potential role of meat cooking practices has not been established yet and could partly explain the current heterogeneity of results among studies. Therefore, we aimed to investigate the association between meat consumption and cooking practices and the risk of CRC in a population-based case–control study.A total of 1671 CRC cases and 3095 controls recruited in Spain between September 2008 and December 2013 completing a food frequency questionnaire with a meat-specific module were included in the analyses. Odds ratios (OR) and confidence intervals (CI) were estimated by logistic regression models adjusted for known confounders. Total meat intake was associated with increased risk of CRC (OR T3–T1 1.41; 95% CI 1.19–1.67; p trend < 0.001), and similar associations were found for white, red and processed/cured/organ meat. Rare-cooked meat preference was associated with low risk of CRC in red meat (ORrare vs. medium 0.66; 95% CI 0.51–0.85) and total meat (ORrare vs. medium 0.56; 95% CI 0.37–0.86) consumers, these associations being stronger in women than in men. Griddle-grilled/barbecued meat was associated with an increased CRC risk (total meat: OR 1.45; 95% CI 1.13–1.87). Stewing (OR 1.25; 95% CI 1.04–1.51) and oven-baking (OR 1.18; 95% CI 1.00–1.40) were associated with increased CRC risk of white, but not red, meat.Our study supports an association of white, red, processed/cured/organ and total meat intake with an increased risk of CRC. Moreover, our study showed that cooking practices can modulate such risk.
Keywords: Colorectal cancer; Meat; Cooking; Epidemiology

The aim of this study is to examine the co-occurrences of low serum ferritin and zinc and anaemia among mothers and their children in two agro-ecological zones of rural Ethiopia.Data were collected from 162 lactating mothers and their breast fed children aged 6–23 months. The data were collected via a structured interview, anthropometric measurements, and blood tests for zinc, ferritin and anaemia. Correlation, Chi-square and multivariable analysis were used to determine the association between nutritional status of mothers and children, and agro-ecological zones.Low serum levels of iron and zinc, anaemia and iron deficiency anaemia were found in 44.4, 72.2, 52.5 and 29.6% of children and 19.8, 67.3, 21.8, 10.5% of mothers, respectively. There was a strong correlation between the micronutrient status of the mothers and the children for ferritin, zinc and anaemia (p < 0.005). Deficiency in both zinc and ferritin and one of the two was observed in 19.1, and 53.7% of the mothers and 32.7 and 46.3%, of their children, respectively. In the 24 h before the survey, 82.1% of mothers and 91.9% of their infants consumed foods that can decrease zinc bioavailability while only 2.5% of mothers and 3.7% of their infants consumed flesh foods.This study shows that micronutrient deficiencies were prevalent among lactating mothers and their children, with variation in prevalence across the agro-ecological zones. This finding calls for a need to design effective preventive public health nutrition programs to address both the mothers’ and their children’s needs.
Keywords: Zinc; Ferritin; Haemoglobin; Mother and children; Ethiopia

In vitro assessment of iron availability from commercial Young Child Formulae supplemented with prebiotics by Tatiana Christides; Julia Clark Ganis; Paul Anthony Sharp (669-678).
Iron is essential for development and growth in young children; unfortunately, iron deficiency (ID) is a significant public health problem in this population. Young Child Formulae (YCF), milk-derived products fortified with iron and ascorbic acid (AA, an enhancer of iron absorption) may be good sources of iron to help prevent ID. Furthermore, some YCF are supplemented with prebiotics, non-digestible carbohydrates suggested to enhance iron bioavailability. The aim of our study was to evaluate iron bioavailability of YCF relative to prebiotic and AA concentrations. We hypothesised that YCF with the highest levels of prebiotics and AA would have the most bioavailable iron.We used the in vitro digestion/Caco-2 cell model to measure iron bioavailability from 4 commercially available YCF with approximately equal amounts of iron, but varying amounts of: AA and the prebiotics fructo- and galacto-oligosaccharides. Caco-2 cell ferritin formation was used as a surrogate marker for iron bioavailability.The YCF with the highest concentration of prebiotics and AA had the highest iron bioavailability; conversely, the YCF with the lowest concentration of prebiotics and AA had the lowest. After the addition of exogenous prebiotics, so that all tested YCF had equivalent amounts, there was no longer a significant difference between YCF iron bioavailability.Our results suggest that ascorbic acid and prebiotics in YCF improve iron bioavailability. Ensuring that iron is delivered in a bioavailable form would improve the nutritional benefits of YCF in relation to ID/IDA amongst young children; therefore, further exploration of our findings in vivo is warranted.
Keywords: Iron; Bioavailability; Prebiotics; Young children; Young Child Formulae; Caco-2

Dietary intake and major food sources of polyphenols in people with type 2 diabetes: The TOSCA.IT Study by M. Vitale; M. Masulli; A. A. Rivellese; E. Bonora; F. Cappellini; A. Nicolucci; S. Squatrito; D. Antenucci; A. Barrea; C. Bianchi; F. Bianchini; L. Fontana; P. Fornengo; F. Giorgino; A. Gnasso; E. Mannucci; A. Mazzotti; R. Nappo; A. P. Palena; P. Pata; G. Perriello; S. Potenziani; R. Radin; L. Ricci; F. Romeo; C. Santini; M. Scarponi; R. Serra; A. Timi; A. A. Turco; M. Vedovato; D. Zavaroni; S. Grioni; G. Riccardi; O. Vaccaro; Angela Albarosa Rivellese; Sara Cocozza; Stefania Auciello; Anna Amelia Turco; Enzo Bonora; Massimo Cigolini; Isabella Pichiri; Corinna Brangani; Elena Tomasetto; Gabriele Perriello; Alessia Timi; Sebastiano Squatrito; Tiziana Sinagra; Sara Longhitano; Vanessa Tropea; Giorgio Ballardini; Anna Carla Babini; Raffaella Ripani; Giovanna Gregori; Maria Dolci; Laura Bruselli; Isabella Salutini; Mary Mori; Fabio Baccetti; Annunziata Lapolla; Giovanni Sartore; Silvia Burlina; Nino Cristiano Chilelli; Raffaella Buzzetti; Chiara Venditti; Stella Potenziani; Angela Carlone; Aldo Galluzzo†; Carla Giordano; Vittoria Torregrossa; Laura Corsi; Giacomo Cuneo; Simona Corsi; Biagio Tizio; Gennaro Clemente; Giuseppe Citro; Maria Natale; Vita Salvatore; Graziano Di Cianni; Emilia Lacaria; Laura Russo; Rossella Iannarelli; Antonella De Gregorio; Filomena Sciarretta; Settimio D’Andrea; Valeria Montani; Emanuela Cannarsa; Katia Dolcetti; Renzo Cordera; Laura Affinito Bonabello; Chiara Mazzucchelli; Carlo Bruno Giorda; Francesco Romeo; Caterina Bonetto; Daniela Antenucci; Maria Pompea Antonia Baldassarre; Ciro Iovine; Rossella Nappo; Ornella Ciano; Elisabetta Dall’Aglio; Giovanni Mancastroppa; Franco Grimaldi; Laura Tonutti; Massimo Boemi; Federica D’Angelo; Sergio Leotta; Lucia Fontana; Davide Lauro; Maria Elena Rinaldi; Mauro Cignarelli; Olga La Macchia; Stefania Fariello; Franco Tomasi; Chiara Zamboni; Nicoletta Dozio; Roberto Trevisan; Cristiana Scaranna; Stefano Del Prato; Roberto Miccoli; Cristina Bianchi; Monia Garofolo; Giuseppe Pugliese; Laura Salvi; Graziela Rangel; Martina Vitale; Roberto Anichini; Anna Tedeschi; Elisa Corsini; Domenico Cucinotta; Antonino Di Benedetto; Loretta Giunta; Maria Concetta Ruffo; Antonio Carlo Bossi; Rita Carpinter; Francesco Dotta; Elena Ceccarelli; Paolo Di Bartolo; Chiara Caselli; Alessandra Luberto; Costanza Santini; Arianna Mazzotti; Giovanni Calbucci; Agostino Consoli; Federica Ginestra; Maria Calabrese; Alessia Zogheri; Lucia Ricci; Francesco Giorgino; Luigi Laviola; Claudia Ippolito; Lucia Tarantino; Angelo Avogaro; Monica Vedovato; Agostino Gnasso; Claudio Carallo; Caterina Scicchitano; Donatella Zavaroni; Stefania Livraga; Paolo Cavallo Perin; Paolo Forrnengo; Tania Prinzis; Salvatore De Cosmo; Antonio Pio Palena; Simonetta Bacci; Edoardo Mannucci; Caterina Lamanna; Pietro Pata; Gabriele Lettina; Antimo Aiello; Angelina Barrea; Carlo Lalli; Maura Scarponi; Ivano Franzetti; Raffaella Radin; Rosalia Serra; Francesca Petrachi; Vincenzo Asprino; Claudio Capra; Massimo Cigolini; Elisa Forte; Stella Potenziani; Giulio Marchesini Reggiani; Gabriele Forlani; Luca Montesi; Natalia Mazzella; Pier Marco Piatti; Lucilla Monti; Michela Stuccillo; Pasquale Auletta; Ettore Petraroli; Giuseppe Capobianco; Geremia Romano; Michele Cutolo; Giosetta De Simone; Gennaro Caiazzo; Peppe Nunziata; Susy Sorrentino; Umberto Amelia; Pasqualino Calatola; Gelsomina Capuano (679-688).
Proper evaluation of polyphenols intake at the population level is a necessary step in order to establish possible associations with health outcomes. Available data are limited, and so far no study has been performed in people with diabetes. The aim of this work was to document the intake of polyphenols and their major food sources in a cohort of people with type 2 diabetes and in socio-demographic subgroups.We studied 2573 men and women aged 50–75 years. Among others, anthropometry was measured by standard protocol and dietary habits were investigated by food frequency questionnaire (EPIC). The intake of polyphenols was evaluated using US Department of Agriculture and Phenol-Explorer databases.The mean total polyphenol intake was 683.3 ± 5.8 mg/day. Non-alcoholic beverages represented the main food source of dietary polyphenols and provided 35.5% of total polyphenol intake, followed by fruits (23.0%), alcoholic beverages (14.0%), vegetables (12.4%), cereal products and tubers (4.6%), legumes (3.7%) and oils (2.1%); chocolate, cakes and nuts are negligible sources of polyphenols in this cohort. The two most important polyphenol classes contributing to the total intake were flavonoids (47.5%) and phenolic acids (47.4%). Polyphenol intake increased with age and education level and decreased with BMI; furthermore, in the northern regions of Italy, the polyphenol intake was slightly, but significantly higher than in the central or southern regions.The study documents for the first time the intake of polyphenols and their main food sources in people with diabetes using validated and complete databases of the polyphenol content of food. Compared with published data, collected in people without diabetes, these results suggest a lower intake and a different pattern of intake in people with diabetes.
Keywords: Polyphenols; Flavonoids; Phenolic acids; Diabetes; Food groups; Diet; Age; BMI; Geographical area; Intake; TOSCA.IT study

Red and processed meat consumption and risk of bladder cancer: a dose–response meta-analysis of epidemiological studies by Alessio Crippa; Susanna C. Larsson; Andrea Discacciati; Alicja Wolk; Nicola Orsini (689-701).
Several epidemiological studies have analyzed the associations between red and processed meat and bladder cancer risk but the shape and strength of the associations are still unclear. Therefore, we conducted a dose–response meta-analysis to quantify the potential association between red and processed meat and bladder cancer risk.Relevant studies were identified by searching the PubMed database through January 2016 and reviewing the reference lists of the retrieved articles. Results were combined using random-effects models.Five cohort studies with 3262 cases and 1,038,787 participants and 8 cases–control studies with 7009 cases and 27,240 participants met the inclusion criteria. Red meat was linearly associated with bladder cancer risk in case–control studies, with a pooled RR of 1.51 (95% confidence interval (CI) 1.13, 2.02) for every 100 g increase per day, while no association was observed among cohort studies (P heterogeneity across study design = 0.02). Based on both case–control and cohort studies, the pooled relative risk (RR) for every 50 g increase of processed meat per day was 1.20 (95% CI 1.06, 1.37) (P heterogeneity across study design = 0.22).This meta-analysis suggests that processed meat may be positively associated with bladder cancer risk. A positive association between red meat and risk of bladder cancer was observed only in case–control studies, while no association was observe in prospective studies.
Keywords: Red meat; Processed meat; Bladder cancer; Dose–response; Meta-analysis

Association of VDBP and CYP2R1 gene polymorphisms with vitamin D status in women with polycystic ovarian syndrome: a north Indian study by Deepa Haldar; Nitin Agrawal; Seema Patel; Pankaj Ramrao Kambale; Kanchan Arora; Aditi Sharma; Manish Tripathi; Aruna Batra; Bhaskar C. Kabi (703-711).
Polycystic ovarian syndrome (PCOS) is the most common endocrine abnormality among women of reproductive age and is usually associated with oligo-ovulation/anovulation, obesity, and insulin resistance. Hypovitaminosis D may also be a primary factor in the initiation and development of PCOS. However, little is known about the role of genetic variation in vitamin D metabolism in PCOS aetiology. Therefore, we studied the genetic polymorphisms of CYP2R1 and vitamin D binding protein (VDBP) in an Indian population.Serum vitamin D was measured by ELISA. Genotyping of VDBP single nucleotide polymorphisms (SNPs) rs7041 (HaeIII; G>T) and rs4588 (StyI; A>C) and CYP2R1 SNP rs2060793 (HinfI; A>G) was carried out by restriction fragment length polymorphism in 50 cases of PCOS that were compared with 50 age-matched healthy women.Vitamin D levels were found to be significantly lower in women with PCOS (p = 0.008) than in age-matched controls. There was no significant difference in genotype frequencies of all three polymorphisms (rs7041, rs4588, and rs2060793) between PCOS and control women. In women with a vitamin D deficiency (<20 ng/ml), the GT allele of the VDBP SNP rs7041 (p value =0.04), the VDBP allelic combination Gc1F/1F (T allele of rs4588 and C allele of rs7041) (p value =0.03), and the GA allele of the CYP2R1 SNP rs2060793 (p = 0.05) were associated with an increased risk of developing PCOS.The present study shows that the GT allele of VDBP SNP rs7041, the VDBP allelic combination (GC1F/1F), and GA allele of CYP2R1 SNP rs2060793 in vitamin D deficient women increase the risk of PCOS.
Keywords: Hypovitaminosis D; PCOS; VDBP; CYP2R1; Single Nucleotide Polymorphisms; PCR; RFLP; Obesity; Insulin Resistance

In a well-characterised community-based prospective study, we aimed to systematically assess the differences in associations of plasma omega-3 and omega-6 fatty acid (FA) status with all-cause mortality when plasma FA status is expressed in absolute concentrations versus relative levels.In a community sample of 564 women aged 25–75 years in Queensland, Australia, baseline plasma phospholipid FA levels were measured using gas chromatography. Specific FAs analysed were eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, total long-chain omega-3 FAs, linoleic acid, arachidonic acid, and total omega-6 FAs. Levels of each FA were expressed in absolute amounts (µg/mL) and relative levels (% of total FAs) and divided into thirds. Deaths were monitored for 17 years and hazard ratios and 95% confidence intervals calculated to assess risk of death according to absolute versus relative plasma FA levels.In total 81 (14%) women died during follow-up. Agreement between absolute and relative measures of plasma FAs was higher in omega-3 than omega-6 FAs. The results of multivariate analyses for risk of all-cause mortality were generally similar with risk tending to inverse associations with plasma phospholipid omega-3 FAs and no association with omega-6 FAs. Sensitivity analyses examining effects of age and presence of serious medical conditions on risk of mortality did not alter findings.The directions and magnitude of associations with mortality of absolute versus relative FA levels were comparable. However, plasma FA expressed as absolute concentrations may be preferred for ease of comparison and since relative units can be deduced from absolute units.
Keywords: Omega-3 fatty acids; Omega-6 fatty acids; Biomarkers; All-cause mortality; Prospective studies

Maternal vitamin D deficiency during pregnancy affects expression of adipogenic-regulating genes peroxisome proliferator-activated receptor gamma (PPARγ) and vitamin D receptor (VDR) in lean male mice offspring by Anthony M. Belenchia; Karen L. Jones; Matthew Will; David Q. Beversdorf; Victoria Vieira-Potter; Cheryl S. Rosenfeld; Catherine A. Peterson (723-730).
Maternal vitamin D deficiency during pregnancy is a widespread issue that may have long-lasting consequences on offspring adiposity. We sought to determine how maternal vitamin D deficiency during the perinatal period would affect offspring adipose tissue development and gene expression.Female C57BL/6 J mice were fed either a vitamin D deficient (VDD) or control diet from 4 weeks before pregnancy (periconception) until 7 days postparturition. Male offspring were weighed and euthanized at 75 days of age (early adult period), at which point serum was collected for biochemical analyses, and perigonadal and subcutaneous white adipose tissue (PGAT and SQAT, respectively) were excised, weighed, then flash-frozen for later histology and analyses of adipogenic gene expression.All adult male offspring were nonobese; there were no significant differences in body weight, adipose pad weight, or adipocyte size. However, VDD-exposed offspring had greater expression of the adipogenic-regulating genes peroxisome proliferator-activated receptor gamma (Pparg) and vitamin D receptor (Vdr).This study suggests that exposure to vitamin D deficiency during the perinatal period can directly affect genes involved in the development of adipose tissue in nonobese offspring. These novel findings invite further investigation into the mechanisms by which maternal vitamin D status during pregnancy affects adipose development and metabolic health of offspring.
Keywords: Vitamin D; Pregnancy; Adipogenesis; Gene expression; Fetal programming; DOHaD

Calcium and vitamin D3 combinations improve fatty liver disease through AMPK-independent mechanisms by Sara Shojaei Zarghani; Hamid Soraya; Mohammad Alizadeh (731-740).
Some research findings suggest that calcium plus vitamin D (VitD) might have a preventive effect on nonalcoholic fatty liver disease development. Moreover, contradictory evidence also exists regarding calcium and VitD deficient diets. This study aimed to evaluate the effect of four different dietary calcium and VitD3 (cholecalciferol) levels on the development of high-fat, high-fructose (HFHFr) diet-induced nonalcoholic fatty liver disease and AMP-activated protein kinase (AMPK) phosphorylation.Thirty male Wistar rats were fed with normal or HFHFr diet containing low calcium (0.2%) and VitD3 (250 IU/kg) (LCD), normal calcium (0.5%) and VtD3 (1000 IU/kg) (CN), high calcium (1.2%) and VitD3 (4000 IU/kg) (HCD) or very high amount of calcium (2.4%) and VitD3 (10,000 IU/kg) (VHCD). After 60 days, anthropometric, metabolic and hepatic parameters were evaluated. The effect of the experimental diets on liver AMPK phosphorylation was also investigated.Rats fed on high calcium plus VitD3 diets, especially VHCD, demonstrated lower adiposity, serum liver enzymes, hepatic lipid accumulation and steatosis. The LCD diet also decreased hepatic lipid content and fatty changes. No evidence indicating the involvement of AMPK in the observed associations was found (P value = 0.51).The results showed high calcium plus VitD3 intakes considerably prevent biochemical and hepatic changes induced by HFHFr diet, probably via an insulin and AMPK-independent pathway. A low intake of these two nutrients was also linked with a significant decrease in HFHFr diet-induced hepatic steatosis.
Keywords: Nonalcoholic fatty liver disease; Calcium; Vitamin D3 ; AMP-activated protein kinase

The association between famine exposure (defined as lengthy and continuous deprivation of food) during early life and type 2 diabetes (T2DM) in adulthood remains controversial. A meta-analysis was performed to better clarify the relation of famine exposure to later T2DM risk.A systematic literature search was performed in PubMed, Web of Science, Embase and China National Knowledge Infrastructure for relevant available articles. The articles were limited to those in English or Chinese from January 1990 to June 2016. Observational studies evaluating the association between famine exposure and T2DM were included. The pooled relative risk (RR) with 95% confidence interval (CI) was used to estimate the effect of famine exposure on T2DM. The I 2 was used to assess heterogeneity, and the random effects model (REM) was adopted as the pooling method.We included 11 published articles with 12391 T2DM cases for this meta-analysis. A significant association of early life famine exposure with increased risk of T2DM was observed (RR = 1.38, 95% CI 1.17–1.63; I 2 = 63.4%; P heterogeneity = 0.002). Compared with the unexposed, the RRs of T2DM were 1.36 (95% CI 1.12–1.65) for fetal-infant exposed and 1.40 (95% CI 0.98–1.99) for childhood exposed. After excluding one article that had a strong effect on heterogeneity, the pooled RR was 1.46 (95% CI 1.27–1.69).The meta-analysis indicates that famine exposure during early life especially fetal-infant exposure may increase the risk of T2DM in adulthood. Measures should be taken to prevent malnutrition during important stages of growth and development to reduce the prevalence of T2DM.
Keywords: Famine exposure; Type 2 diabetes; Risk; Meta-analysis

Determinants of folate and vitamin B12 plasma levels in the French E3N-EPIC cohort by Jordi de Batlle; Marco Matejcic; Veronique Chajes; Hortensia Moreno-Macias; Amina Amadou; Nadia Slimani; David G. Cox; Françoise Clavel-Chapelon; Guy Fagherazzi; Isabelle Romieu (751-760).
Impaired B vitamin status has been identified as a risk factor for major chronic diseases. This study aims at examining the determinants of plasma folate and vitamin B12 concentrations, considering lifestyle factors and MTHFR polymorphisms.A total of 988 women aged 40–65 years from the French E3N cohort were investigated. Intakes of folate and vitamin B12 were assessed using food frequency questionnaires, and plasma concentrations were measured by microbiological assay. Dietary scores were computed to summarize folate and vitamin B12 dietary sources. MTHFR-C677T and MTHFR-A1298C were determined by Kaspar assay. Pearson’s partial correlation coefficients and multivariable linear regression models were used to assess correlations between main determinants and plasma folate and vitamin B12 levels.The partial correlation coefficient between dietary intakes and plasma folate was 0.19 (p value <0.001) and 0.08 (p value = 0.008) for vitamin B12. Dietary scores were the main determinant of B vitamin plasma concentrations with a percent change per unit increase of 12.64% (p value <0.001) for folate and 7.6% (p value <0.001) for vitamin B12. Homozygous (T/T) or heterozygous (C/T) women for MTHFR-C677T had lower plasma folate concentrations [C/T: −6.48% (p value = 0.038) and T/T: −15.89% (p value <0.001)] compared to women carrying the C/C genotype. Other determinants of B vitamin plasma concentration include: smoking status for folate, and age and hormone replacement therapy for vitamin B12.We confirmed previous findings on the role of diet as main determinant of folate and vitamin B12 plasma concentrations. However, the impact of genetic polymorphisms and lifestyle factors on plasma B vitamin concentrations should not be neglected.
Keywords: Folate; Vitamin B12; Dietary intake; Biomarkers; MTHFR polymorphisms

Rice bran protein hydrolysates attenuate diabetic nephropathy in diabetic animal model by Kampeebhorn Boonloh; Eun Soo Lee; Hong Min Kim; Mi Hye Kwon; You Mi Kim; Patchareewan Pannangpetch; Bunkerd Kongyingyoes; Upa Kukongviriyapan; Supawan Thawornchinsombut; Eun Young Lee; Veerapol Kukongviriyapan; Choon Hee Chung (761-772).
Diabetic nephropathy (DN) is an important microvascular complication of uncontrolled diabetes. The features of DN include albuminuria, extracellular matrix alterations, and progressive renal insufficiency. Rice bran protein hydrolysates (RBPs) have been reported to have antihyperglycemic, lipid-lowering, and anti-inflammatory effects in diabetic rats. Our study was to investigate the renoprotective effects of RBP in diabetic animals and mesangial cultured cells.Eight-week-old male db/m and db/db mice were orally treated with tap water or RBP (100 or 500 mg/kg/day) for 8 weeks. At the end of the experiment, diabetic nephropathy in kidney tissues was investigated for histological, ultrastructural, and clinical chemistry changes, and biomarkers of angiogenesis, fibrosis, inflammation, and antioxidant in kidney were analyzed by Western blotting. Protection against proangiogenic proteins and induction of cytoprotection by RBP in cultured mesangial cells was evaluated.RBP treatment improved insulin sensitivity, decreased elevated fasting serum glucose levels, and improved serum lipid levels and urinary albumin/creatinine ratios in diabetic mice. RBP ameliorated the decreases in podocyte slit pore numbers, thickening of glomerular basement membranes, and mesangial matrix expansion and suppressed elevation of MCP-1, ICAM-1, HIF-1α, VEGF, TGF-β, p-Smad2/3, and type IV collagen expression. Moreover, RBP restored suppressed antioxidant Nrf2 and HO-1 expression. In cultured mesangial cells, RBP inhibited high glucose-induced angiogenic protein expression and induced the expression of Nrf2 and HO-1.RBP attenuates the progression of diabetic nephropathy and restored renal function by suppressing the expression of proangiogenic and profibrotic proteins, inhibiting proinflammatory mediators, and restoring the antioxidant and cytoprotective system.
Keywords: Diabetic nephropathy; Angiogenesis; Fibrosis; Rice bran; Insulin resistance; Inflammation

Type of sweet flavour carrier affects thyroid axis activity in male rats by Ewelina Pałkowska-Goździk; Anna Bigos; Danuta Rosołowska-Huszcz (773-782).
Non-nutritive sweeteners are the most widely used food additives worldwide. However, their metabolic outcomes are still a matter of controversy and their effect on the thyroid activity, a key regulator of metabolism, has not been previously studied. Therefore, we aim to determine the influence of the sweet type flavour carrier on selected parameters of thyroid axis activity.Male Sprague–Dawley rats (n = 105) were divided into 3 groups fed ad libitum for three weeks isocaloric diets (3.76 ± 0.5 kcal/g): two with the same sweet flavour intensity responded to 10% of sucrose (with sucrose–SC—and sucralose—SU) and one non-sweet diet (NS). To evaluate the post-ingested effects, animals were euthanised at fast and 30, 60, 120, 180 min after meal.The results obtained indicate that both the presence and the type of sweet taste flavour carrier affect thyroid axis activity both at fasting and postprandial state. Compared to diet with sucrose which stimulates thyroid axis activity, sucralose addition diminishes thyroid hormone synthesis as thyroid peroxidase (TPO) activity, plasma thyroxine (T4), and triiodothyronine (T3) concentration was lower than in SC and NS while in non-sweet diet the lowest level of hepatic deiodinase type 1 (DIO1) and the highest reverse T3 (rT3) level indicate on altered thyroid hormone peripheral metabolism.Both the presence and the type of sweet flavour carrier have a significant impact on thyroid axis activity. Our findings suggest that this organochlorine sweetener is metabolically active and might exacerbate metabolic disorders via an adverse effect on thyroid hormone metabolism.
Keywords: Thyroid hormones; Sucrose; Non-nutritive sweeteners; Sucralose

Low vitamin D deficiency in Irish toddlers despite northerly latitude and a high prevalence of inadequate intakes by Carol ní Chaoimh; Elaine K. McCarthy; Jonathan O’B. Hourihane; Louise C. Kenny; Alan D. Irvine; Deirdre M. Murray; Mairead E. Kiely (783-794).
While reports of inadequate vitamin D intakes among young children are widespread, data on the prevalence of vitamin D deficiency are inconsistent. We aimed to quantify vitamin D intake and serum 25-hydroxyvitamin D [25(OH)D] concentrations in children aged 2 years in the prospective Cork BASELINE Birth Cohort Study.Serum 25(OH)D was analysed using UPLC-MS/MS in 741 children living in Cork, Ireland (51°N). Two-day weighed food diaries were collected in 467 children, and 294 provided both a blood sample and a food diary.Mean (SD) 25(OH)D concentrations were 63.4 (20.4) nmol/L [winter: 54.5 (19.9), summer: 71.2 (17.5)]. The prevalence of vitamin D deficiency (<30 nmol/L) was 4.6, and 26.7% were <50 nmol/L [45.2% during winter (November–April) and 10.4% in summer (May–October)]. With a mean (SD) vitamin D intake of 3.5 (3.1) µg/day, 96% had intakes below 10 µg/day, the current IOM estimated average requirement and the SACN safe intake value for this age group. After adjustment for season, vitamin D intake (µg/day) was associated with higher 25(OH)D concentrations [adjusted estimate (95% CI) 2.5 (1.9, 3.1) nmol/L]. Children who did not consume vitamin D-fortified foods or supplements had very low vitamin D intakes (1.2 (0.9) µg/day), and during winter, 12 and 77% were <30 and <50 nmol/L, respectively, compared with 6 and 44% of fortified food consumers.There was a high prevalence of low vitamin D status during winter, especially among children who did not consume fortified foods or nutritional supplements. Our data indicate the need for dietary strategies to increase vitamin D intakes in this age group. This report provides further evidence that DRVs for vitamin D should be based on experimental data in specific population groups and indicates the need for dose–response RCTs in young children.
Keywords: Vitamin D intake; 25-Hydroxyvitamin D; Fortified foods; Supplements; Young children

Effects of a diet rich in arabinoxylan and resistant starch compared with a diet rich in refined carbohydrates on postprandial metabolism and features of the metabolic syndrome by Anne Grethe Schioldan; Søren Gregersen; Stine Hald; Ann Bjørnshave; Mette Bohl; Bolette Hartmann; Jens Juul Holst; Hans Stødkilde-Jørgensen; Kjeld Hermansen (795-807).
Low intake of dietary fibre is associated with the development of type 2 diabetes. Dyslipidaemia plays a key role in the pathogenesis of type 2 diabetes. Knowledge of the impact of dietary fibres on postprandial lipaemia is, however, sparse. This study aimed in subjects with metabolic syndrome to assess the impact on postprandial lipaemia and features of the metabolic syndrome of a healthy carbohydrate diet (HCD) rich in cereal fibre, arabinoxylan and resistant starch compared to a refined-carbohydrate western-style diet (WSD).Nineteen subjects completed the randomised, crossover study with HCD and WCD for 4-week. Postprandial metabolism was evaluated by a meal-challenge test and insulin sensitivity was assessed by HOMA-IR and Matsuda index. Furthermore, fasting cholesterols, serum-fructosamine, circulating inflammatory markers, ambulatory blood pressure and intrahepatic lipid content were measured.We found no diet effects on postprandial lipaemia. However, there was a significant diet × statin interaction on total cholesterol (P = 0.02) and LDL cholesterol (P = 0.002). HCD decreased total cholesterol (−0.72 mmol/l, 95% CI (−1.29; −0.14) P = 0.03) and LDL cholesterol (−0.61 mmol/l, 95% CI (−0.86; −0.36) P = 0.002) compared with WSD in subjects on but not without statin treatment. We detected no other significant diet effects.In subjects with metabolic syndrome on statins a 4-week diet rich in arabinoxylan and resistant starch improved fasting LDL and total cholesterol compared to subjects not being on statins. However, we observed no diet related impact on postprandial lipaemia or features of the metabolic syndrome. The dietary fibre x statin interaction deserves further elucidation.
Keywords: Metabolic syndrome; Dietary fibre; Lipaemia; Insulin resistance

Iodine intake as a risk factor for BRAF mutations in papillary thyroid cancer patients from an iodine-replete area by Hye Jeong Kim; Hyeong Kyu Park; Dong Won Byun; Kyoil Suh; Myung Hi Yoo; Yong-Ki Min; Sun Wook Kim; Jae Hoon Chung (809-815).
Both deficient and excessive iodine intake leads to thyroid disease, which shows U-shaped curves. Our previous study showed that a relatively low [urinary iodine concentration (UIC) <300 μg/L] and extremely excessive (UIC ≥ 2500 μg/L) iodine intake were associated with thyroid cancer in Korea, an iodine-replete area. Papillary thyroid cancer (PTC) accounts for more than 97 % of thyroid cancer and 80% or more PTC cases harbor the BRAF mutation in Korea. We aimed to investigate the relationship between iodine intake and the prevalence of the BRAF mutation in PTC in Korea.UIC was measured by inductively coupled plasma mass spectrometry. The BRAF mutation was detected using both allele-specific polymerase chain reaction and mutant enrichment with 3′-modified oligonucleotide sequencing. Risk factors for the occurrence of BRAF mutations in PTC were evaluated using multivariate logistic regression models.The median UIC in all patients with PTC was 287 μg/L (range from 7 to 7, 426 μg/L). Nearly half of the patients (102/215, 47%) belonged to the excessive iodine intake category (UIC ≥ 300 μg/L) according to the WHO iodine recommendations. The frequency of BRAF mutations was lowest in the 300–499 μg/L UIC group; it was significantly different compared to the relatively low (UIC < 300 μg/L) and more than excessive (UIC ≥ 500 μg/L) iodine intake groups. UIC was an independent predictor for BRAF mutations in PTC. The multivariate-adjusted odds ratios (95% confidence intervals) in the relatively low and more than excessive iodine intake groups for the BRAF mutation were 4.761 (1.764–12.850) and 6.240 (2.080–18.726), respectively, compared to the 300–499 μg/L UIC group.Relatively low iodine intake and more than excessive iodine intake seem to be significant risk factors for the occurrence of BRAF mutations in the thyroid and, therefore, may be risk factors for the development of PTC in an iodine-replete area.
Keywords: Iodine; BRAF mutation; Papillary thyroid cancer

Euterpe oleracea Mart. seed extract protects against renal injury in diabetic and spontaneously hypertensive rats: role of inflammation and oxidative stress by Viviane da Silva Cristino Cordeiro; Graziele Freitas de Bem; Cristiane Aguiar da Costa; Izabelle Barcellos Santos; Lenize Costa Reis Marins de Carvalho; Dayane Teixeira Ognibene; Ana Paula Machado da Rocha; Jorge José de Carvalho; Roberto Soares de Moura; Angela Castro Resende (817-832).
Euterpe oleracea Mart. (açaí) seed extract (ASE), through its anti-hypertensive, antioxidant and anti-inflammatory properties, may be useful to treat or prevent human diseases. Several evidences suggest that oxidative stress and inflammation contribute to the pathogenesis of diabetic nephropathy; therefore, we tested the hypothesis that ASE (200 mg/kg−1day−1) prevents diabetes and hypertension-related oxidative stress and inflammation, attenuating renal injury.Male rats with streptozotocin (STZ)-induced diabetes (D), and spontaneously hypertensive rats with STZ-induced diabetes (DH) were treated daily with tap water or ASE (D + ASE and DH + ASE, respectively) for 45 days. The control (C) and hypertensive (H) animals received water.The elevated serum levels of urea and creatinine in D and DH, and increased albumin excretion in HD were reduced by ASE. Total glomeruli number in D and DH, were increased by ASE that also reduced renal fibrosis in both groups by decreasing collagen IV and TGF-β1 expression. ASE improved biomarkers of renal filtration barrier (podocin and nephrin) in D and DH groups and prevented the increased expression of caspase-3, IL-6, TNF-α and MCP-1 in both groups. ASE reduced oxidative damage markers (TBARS, carbonyl levels and 8-isoprostane) in D and DH associated with a decrease in Nox 4 and p47 subunit expression and increase in antioxidant enzyme activity in both groups (SOD, catalase and GPx).ASE substantially reduced renal injury and prevented renal dysfunction by reducing inflammation, oxidative stress and improving the renal filtration barrier, providing a nutritional resource for prevention of diabetic and hypertensive-related nephropathy.
Keywords: Euterpe oleracea Mart; Diabetes; Hypertension; Nephropathy; Oxidative stress; Inflammation

In the original publication, one of the author’s name was misspelled. The correct name is “Salma Mahmoodianfard”. The original article was corrected.Overweight and obesity are major public health concerns worldwide which are associated with a low-grade chronic inflammation. Dietary fiber as an important component of diet could be effective in controlling weight and inflammatory factors. The present study aimed to compare the effects of rice husk powder and rice bran on inflammatory factors along with an energy-restricted diet in overweight and obese adults.In this randomized trial, 105 eligible individuals were assigned to one of the three energy-restricted diet groups receiving; rice bran (n = 35), rice husk powder (n = 35), and control group (n = 35) for 12 weeks. Demographic data, dietary intake, anthropometric indices and inflammatory factors (serum levels of IL-6 and hs-CRP) were measured at baseline and at the end of the study.Weight, BMI and waist circumference reduced significantly in all groups after 12 weeks of study (P < 0.01 for all). However, pre- and post-measure differences between groups were not significant. Moreover, serum levels of hs-CRP and IL-6 were not significantly different between participants in the rice bran or rice husk groups. However, the reduction in serum levels of hs-CRP in rice husk (mean change = − 0.14 ± 0.05 µg/ml) and rice bran (mean change = − 0.13 ± 0.03 µg/ml) was significantly higher when compared to the control group (mean change = − 0.03 ± 0.02 µg/ml) (P < 0.05 for both groups). The same pattern was found when changes in IL-6 serum levels of participants in rice husk (mean change = − 0.48 ± 0.11 pg/ml) and rice bran (mean change = − 0.57 ± 0.13 pg/ml) groups were compared to the control group (mean change= − 0.19 ± 0.07 pg/ml) (P < 0.05 for both groups).The results of this study showed positive effects of rice bran and rice husk powder supplementation, combined with an energy-restricted diet, on inflammatory markers among overweight and obese adults.
Keywords: Rice bran; Rice husk; Overweight; Obesity; IL-6; Hs-CRP

In the original publication, one of the author’s name was misspelled. The correct name is “Salma Mahmoodianfard”. The original article was corrected.