European Journal of Nutrition (v.57, #3)

Challenges in the assessment of total fluid intake in children and adolescents: a discussion paper by Janet Warren; Isabelle Guelinckx; Barbara Livingstone; Nancy Potischman; Michael Nelson; Emma Foster; Bridget Holmes (43-51).
In the original publication of the article, a mistake was introduced in affiliation of Dr. Michael Nelson.In recent years, evidence has emerged about the importance of healthy fluid intake in children for physical and mental performance and health, and in the prevention of obesity. Accurate data on water intake are needed to inform researchers and policymakers and for setting dietary reference values. However, to date, there are few published data on fluid or water intakes in children. This is due partly to the fact that drinking water is not always reported in dietary surveys. The aim of this paper is to review the current status of the literature and highlight the challenges of assessing total fluid intake in children and adolescents.From the dietary assessment literature it is apparent that children present unique challenges to assessing intake due to ongoing cognitive capacity development, limited literacy skills, difficulties in estimating portion sizes and multiple caregivers during any 1 day making it difficult to track intakes. As such, many issues should be considered when assessing total fluid intakes in children or adolescents. Various methods to assess fluid intakes exist, each with its own strengths and weaknesses; the ultimate choice of method depends on the research question and resources available. Based on the literature review, it is apparent that if the research focus is to assess only fluid intake, a fluid-specific method, such as a diary or record, appears to be a feasible approach to provide an accurate estimate of intakes.
Keywords: Dietary assessment; Beverages; Fluid intake; Water; Hydration; Children

Fluid intake of Latin American children and adolescents: results of four 2016 LIQ.IN 7 National Cross-Sectional Surveys by J. Gandy; H. Martinez; E. Carmuega; J. L. Arredondo; C. Pimentel; L. A. Moreno; S. A. Kavouras; J. Salas-Salvadó (53-63).
The primary aim of this survey was to report total fluid intake (TFI) and different fluid types for children (4–9 years) and adolescents (10–17 years) in Mexico, Brazil, Argentina and Uruguay. The second aim was to compare TFI with the adequate intake (AI) of water from fluids as recommended by the USA Institute of Medicine.Data were collected using a validated liquid intake 7-day record (Liq.In 7 ). Participants’ characteristics, including age, sex and anthropometric measurements were recorded.A total of 733 children and 933 adolescents were recruited. Over 75% of children in Uruguay met the IOM’s recommended intake. Fewer children in Argentina (64–72%) and Brazil (41–50%) obtained AI and the lowest values were recorded in Mexico (33–44%), where 16% of boys and 14% girls drank 50% or less of the AI. More adolescents in Argentina (42%) met the AIs than other countries; the lowest was in Mexico (28%). Children and adolescents in Mexico and Argentina drank more sugar sweetened beverages than water.Large numbers of children and adolescents did not meet AI recommendations for TFI, raising concerns about their hydration status and potential effects on mental and physical well-being. Given the negative effects on children’s health, the levels of SSB consumption are worrying.
Keywords: Beverages; Fluid intake; Water; Hydration; 7 ; Children; Adolescents; Mexico; Brazil; Uruguay; Argentina

Fluid intake of Latin American adults: results of four 2016 Liq.In7 national cross-sectional surveys by H. Martinez; C. Morin; J. Gandy; E. Carmuega; J. L. Arredondo; C. Pimentel; L. A. Moreno; S. A. Kavouras; J. Salas-Salvadó; I. Guelinckx (65-75).
To report total fluid intake (TFI) and the intake of different fluid types in adults (≥ 18 years old) from Mexico, Argentina, Brazil and Uruguay. To compare intakes between countries and with recommended adequate intake (AI) of water from fluids.Cross-sectional data were collected using a validated liquid intake 7-day record (Liq.In 7 ) in populations from Argentina (n = 1089), Brazil (n = 477), Mexico (n = 1677) and Uruguay (n = 554). Population characteristics, including age, gender, body mass index and socioeconomic level were recorded. Mean TFI was compared with the AI of water from fluids set by the USA Institute of Medicine.The lowest TFI was recorded in Mexican women (1748 mL/day) and the highest in Argentinean men (2318 mL/day). Median daily TFI was significantly different between countries; Uruguay and Argentina had higher values than Mexico and Brazil. In the former, plain water contributed to only 25% of TFI, the remainder being predominantly from hot beverages. Approximately, a third of adults did not drink enough fluid to meet the recommended AI. High SSB consumption was reported, which was significantly different between countries (p < 0.05), the highest being in Mexico (median 25–75th percentiles): 531 (300–895 mL/day.This survey highlights the need to increase water consumption and reduce SSB intake in this region to avoid potential associated health risks. These findings may be useful information in monitoring public health policy strategies.
Keywords: Beverages; Fluid intake; Hydration; Liq.In7 ; Water; Argentina; Brazil; Mexico; Uruguay

Fluid intake in urban China: results of the 2016 Liq.In 7 national cross-sectional surveys by N. Zhang; C. Morin; I. Guelinckx; L. A. Moreno; S. A. Kavouras; J. Gandy; H. Martinez; J. Salas-Salvadó; G. Ma (77-88).
To describe total fluid intake (TFI) and types of fluid consumed in urban China by age, gender, regions and city socioeconomic status relative to the adequate intakes (AI) set by the Chinese Nutrition Society.In 2016, participants aged 4–9, 10–17 and 18–55 years were recruited via a door-to-door approach in 27 cities in China. In total, 2233 participants were included. The volumes and sources of TFI were collected using the Liq.In 7 record, assisted by a photographic booklet of standard fluid containers.The mean daily TFI among children, adolescents and adults were 966, 1177 and 1387 mL, respectively. In each age group, TFI was significantly higher in male vs female (981 vs 949, 1240 vs 1113, 1442 vs 1332; mL). Approximately 45, 36 and 28% of children, adolescents and adults reached the AI. Although plain water was the highest contributor to TFI, the contribution of sugar sweetened beverages (SSB) was ranked in the top three together with water and milk and derivatives. Approximately 27, 48 and 47% of children, adolescents and adults consumed more than one serving of SSB per day, respectively.A relatively large proportion of participants did not drink enough to meet the AI in urban China. Many children, adolescents and adults consumed more than one serving of SSB per day. A majority of children, adolescents and adults in the study population do not meet both quantitative and qualitative fluid intake requirements, and signal socioeconomic disparities.
Keywords: Fluid types; Adequate water intake; Sugar sweetened beverages; Healthy hydration

Fluid intake of children, adolescents and adults in Indonesia: results of the 2016 Liq.In7 national cross-sectional survey by P. W. Laksmi; C. Morin; J. Gandy; L. A. Moreno; S. A. Kavouras; H. Martinez; J. Salas-Salvadó; I. Guelinckx (89-100).
To report daily total fluid intake (TFI) and fluid types in Indonesia according to age, sex, socio-economic status (SES) and geographic region, and compare TFI with the Indonesian adequate fluid intake (AI) recommendations.Data were collected in 32 cities over nine regions from children (4–9 years, n = 388), adolescents, (10–17 years, n = 478) and adults (18–65 years, n = 2778) using a fluid intake 7-day record (Liq.In7); socio-economic status was also recorded. The 7-day mean TFIs were compared with the AI of water set by the Ministry of Health of the Republic of Indonesia.Total median fluid intakes for all age groups exceeded 2000 mL/day. At population level, TFI was associated with household income (P < 0.001), education (P < 0.001) and Indonesian geographical regions (P < 0.001). More than 67% of participants met the AI of water from fluids. A higher percentage of children and adolescents met the AI (78 and 80%, respectively), compared with adults (72%). Drinking water was the main contributor to TFI in all age groups (76–81%). Sugar-sweetened beverages (SSB) were consumed by 62% children, 72% adolescents and 61% of adults. An SSB intake ≥ 1 serving per day was observed for 24% children, 41% adolescents and 33% adults.A high percentage of the population drank enough to meet the AI of water from fluids. Water was the most frequently consumed drink; however, many participants consumed at least one serving of SSB per day. This study provides data to help direct targeted intervention programs.
Keywords: Beverages; Fluid intake; Hydration; Indonesia; Liq.In7 ; Water

A comparison of drinking behavior using a harmonized methodology (Liq.In 7 ) in six countries by C. Morin; J. Gandy; L. A. Moreno; S. A. Kavouras; H. Martinez; J. Salas-Salvadó; I. Guelinckx (101-112).
To assess drinking occasions (volume and type) according to consumption with food in or outside meals, and location, for six countries.A total of 10,521 participants aged 4–65 years from Argentina, Brazil, China, Indonesia, Mexico and Uruguay completed a validated 7-day fluid intake record. For each drinking event, the volume consumed, the fluid type, the location of intake, and whether the drink was accompanied by food (meal or snack) or not, was recorded.Similar drinking behaviors were found in Mexico and Argentina; fluid intake during meals was 48 and 45% of total fluid intake (TFI), respectively. In Brazil (55%), Indonesia (58%) and China (66%) most fluid was consumed without food. In Uruguay, 34% of TFI was with a main meal, 31% with food between meals and 35% without food. Indonesia had the highest median (25–75th percentile) TFI; 2520 (1750–3347) mL/day, and China the lowest 1138 (818–3347) mL/day. Water was consumed with meals for 37% of Chinese and 87% of Indonesian participants, while the four Latin-American American countries showed a preference for sweet drinks; 54% in Mexico, 67% in Brazil, 55% in Argentina and 59% in Uruguay. Diversity in fluid type was noted when drinking with food between meals. Apart from China, most drinking occasions (> 75%) occurred at home.Three distinct drinking behaviors were identified, namely, drinking with meals, drinking as a stand-alone activity, and a type of ‘grazing’ (i.e., frequent drinks throughout the day) behavior. Most drinking occasions occurred at home.
Keywords: Beverages; Fluid intake; Water; Hydration; Liq.In 7 ; Behavior

Fluid intake patterns of children and adolescents: results of six Liq.In7 national cross-sectional surveys by C. Morin; J. Gandy; R. Brazeilles; L. A. Moreno; S. A. Kavouras; H. Martinez; J. Salas-Salvadó; J. Bottin; Isabelle Guelinckx (113-123).
This study aimed to identify and characterize patterns of fluid intake in children and adolescents from six countries: Argentina, Brazil, China, Indonesia, Mexico and Uruguay.Data on fluid intake volume and type amongst children (4–9 years; N = 1400) and adolescents (10–17 years; N = 1781) were collected using the validated 7-day fluid-specific record (Liq.In7 record). To identify relatively distinct clusters of subjects based on eight fluid types (water, milk and its derivatives, hot beverages, sugar-sweetened beverages (SSB), 100% fruit juices, artificial/non-nutritive sweetened beverages, alcoholic beverages, other beverages), a cluster analysis (partitioning around k-medoids algorithm) was used. Clusters were then characterized according to their socio-demographics and lifestyle indicators.The six interpretable clusters identified were: low drinkers–SSB (n 523), low drinkers–water and milk (n 615), medium mixed drinkers (n 914), high drinkers–SSB (n 513), high drinkers–water (n 352) and very high drinkers–water (n 264). Country of residence was the dominant characteristic, followed by socioeconomic level, in all six patterns.This analysis showed that consumption of water and SSB were the primary drivers of the clusters. In addition to country, socio-demographic and lifestyle factors played a role in determining the characteristics of each cluster. This information highlights the need to target interventions in particular populations aimed at changing fluid intake behavior and improving health in children and adolescents.
Keywords: Beverages; Fluid intake; Water; Hydration; Liq.In7 ; Children; Adolescents; Clustering analysis

Reduced energy availability: implications for bone health in physically active populations by Maria Papageorgiou; Eimear Dolan; Kirsty J. Elliott-Sale; Craig Sale (847-859).
The present review critically evaluates existing literature on the effects of short- and long-term low energy availability (EA) on bone metabolism and health in physically active individuals.We reviewed the literature on the short-term effects of low EA on markers of bone metabolism and the long-term effects of low EA on outcomes relating to bone health (bone mass, microarchitecture and strength, bone metabolic markers and stress fracture injury risk) in physically active individuals.Available evidence indicates that short-term low EA may increase markers of bone resorption and decrease markers of bone formation in physically active women. Bone metabolic marker responses to low EA are less well known in physically active men. Cross-sectional studies investigating the effects of long-term low EA suggest that physically active individuals who have low EA present with lower bone mass, altered bone metabolism (favouring bone resorption), reduced bone strength and increased risk for stress fracture injuries.Reduced EA has a negative influence on bone in both the short- and long-term, and every effort should be made to reduce its occurrence in physically active individuals. Future interventions are needed to explore the effects of long-term reduced EA on bone health outcomes, while short-term low EA studies are also required to give insight into the pathophysiology of bone alterations.
Keywords: Energy availability; Physically active individuals; Female Athlete Triad; Relative Energy Deficiency in Sports; Bone health; Bone metabolic markers

Influence of gut microbiota on the development and progression of nonalcoholic steatohepatitis by Fabiana de Faria Ghetti; Daiane Gonçalves Oliveira; Juliano Machado de Oliveira; Lincoln Eduardo Villela Vieira de Castro Ferreira; Dionéia Evangelista Cesar; Ana Paula Boroni Moreira (861-876).
Nonalcoholic steatohepatitis (NASH) is characterized by the presence of steatosis, inflammation, and ballooning degeneration of hepatocytes, with or without fibrosis. The prevalence of NASH has increased with the obesity epidemic, but its etiology is multifactorial. The current studies suggest the role of gut microbiota in the development and progression of NASH. The aim is to review the studies that investigate the relationship between gut microbiota and NASH. These review also discusses the pathophysiological mechanisms and the influence of diet on the gut–liver axis.The available literature has proposed mechanisms for an association between gut microbiota and NASH, such as: modification energy homeostasis, lipopolysaccharides (LPS)–endotoxemia, increased endogenous production of ethanol, and alteration in the metabolism of bile acid and choline. There is evidence to suggest that NASH patients have a higher prevalence of bacterial overgrowth in the small intestine and changes in the composition of the gut microbiota. However, there is still a controversy regarding the microbiome profile in this population. The abundance of Bacteroidetes phylum may be increased, decreased, or unaltered in NASH patients. There is an increase in the Escherichia and Bacteroides genus. There is depletion of certain taxa, such as Prevotella and Faecalibacterium.Although few studies have evaluated the composition of the gut microbiota in patients with NASH, it is observed that these individuals have a distinct gut microbiota, compared to the control groups, which explains, at least in part, the genesis and progression of the disease through multiple mechanisms. Modulation of the gut microbiota through diet control offers new challenges for future studies.
Keywords: Nonalcoholic fatty liver disease; Dysbiosis; Steatohepatitis; Gut microbiota; Microbiome

The aim of the current meta-analysis was to investigate the effect of increasing dietary ALA intake on the blood concentration of inflammatory markers including tumor necrosis factor (TNF), interleukin 6 (IL-6), C-reactive protein (CRP), soluble intercellular adhesion molecule-1 (sICAM-1), and soluble vascular cell adhesion molecule-1 (sVCAM-1) in adults.After a systemic search on PubMed, Embase, and Cochrane library and bibliographies of relevant articles, 25 randomized controlled trials that met the inclusion criteria were identified.No significant effect of dietary ALA supplementation was observed on TNF (SMD: −0.03, 95% CI −0.36 to 0.29), IL-6 (SMD: −0.17, 95% CI −0.46 to 0.12), CRP (SMD: −0.06, 95% CI −0.24 to 0.12), sICAM-1 (SMD: −0.06, 95% CI −0.26 to 0.13), and sVCAM-1 (SMD: −0.24, 95% CI −0.56 to 0.09). Subgroup analysis revealed that increasing dietary ALA tends to elevate CRP concentration in healthy subjects. However, the null effect of ALA supplementation on other inflammatory markers was not changed in various subgroups, indicating that the results are stable. Meta-regression results revealed a negative relationship between the effect size on CRP and its baseline concentration. No significant publication bias was observed for all inflammatory markers as suggested by funnel plot and Begg’s test.Our meta-analysis did not find any beneficial effect of ALA supplementation on reducing inflammatory markers including TNF, IL-6, CRP, sICAM-1, and sVCAM-1. However, in healthy subjects, ALA supplementation might increase CRP concentration.
Keywords: Alpha-linolenic acid; Inflammatory markers; Randomized controlled trials; Meta-analysis

Fish oil supplementation attenuates neuroinflammation and alleviates depressive-like behavior in rats submitted to repeated lipopolysaccharide by Ruili Dang; Xueyuan Zhou; Mimi Tang; Pengfei Xu; Xiaoxue Gong; Yuanyuan Liu; Hongxiao Jiao; Pei Jiang (893-906).
Depression is frequently associated with inflammation, whereas omega-3 polyunsaturated fatty acids (PUFAs) primarily found in fish oil possess anti-inflammatory properties. Although converging studies suggest an antidepressant effect of PUFAs, there is limited evidence directly linking the neuro-immune modulating features of PUFAs to the antidepressant actions.Therefore, we assessed the effects of fish oil (FO) supplementation on behavioral changes, inflammatory cytokine expression and oxidative reactions in frontal cortex and hippocampus of rats following repeated peripheral immune challenge by lipopolysaccharide (LPS) for 2 weeks (500 μg/kg every other day).Repeated LPS administration induced the rats to a depressive-like state and increased mRNA expression of pro-inflammatory cytokines, including 1L-1β, 1L-6 and TNF-α, in frontal cortex and hippocampus. FO supplementation attenuated the LPS-induced abnormal behavior and brain inflammatory response. Concurrent with the antidepressant action, FO also reduced LPS-induced oxidative reactions and neural apoptosis in the rat brain, as evidenced by decreased malondialdehyde (MDA) production, increased catalase activities and inhibited pro-apoptotic protein Bax mRNA expression. In addition, FO inhibited activation of NF-κB and iNOS induced by LPS. Interestingly, we found FO suppressed the activation of the inflammasome NLRP3 and ionotropic purinergic receptor P2X7R evoked by LPS, suggesting a potential anti-inflammatory mechanism for PUFAs. Besides, FO also restored the LPS-induced neurochemical disturbance, especially the balance between serotonin and kynurenine branches of tryptophan metabolism, which is tightly associated with depression.These findings provide novel insights into the antidepressant action of PUFAs and further strengthen the link between inflammation and depression.
Keywords: ω-3 Polyunsaturated fatty acids; Lipopolysaccharide; Neuroinflammation; Depression; P2X7R; Neurotransmitters

Effects of tryptophan-containing peptides on angiotensin-converting enzyme activity and vessel tone ex vivo and in vivo by Sherif Khedr; Andreas Deussen; Irakli Kopaliani; Birgit Zatschler; Melanie Martin (907-915).
Over-activation of the renin-angiotensin axis and worsening of vascular function are critical contributors to the development of hypertension. Therefore, inhibition of angiotensin-converting enzyme (ACE), a key factor of the renin-angiotensin axis, is a first line treatment of hypertension. Besides pharmaceutical ACE inhibitors, some natural peptides have been shown to exert ACE-inhibiting properties with antihypertensive effects and potentially beneficial effects on vascular function. In this study, the ACE-inhibiting potential and effects on vascular function of tryptophan-containing peptides were evaluated.The ACE inhibitory action and stability of tryptophan-containing peptides was tested in endothelial cells—a major source of whole body ACE activity. Furthermore, the efficacy of peptides on vascular ACE activity, as well as vessel tone was assessed both ex vivo and in vivo.In human umbilical vein endothelial cells (HUVEC), isoleucine-tryptophan (IW) had the highest ACE inhibitory efficacy, followed by glutamic acid-tryptophan (EW) and tryptophan-leucine (WL). Whereas none of the peptides affected basal vessel tone (rat aorta), angiotensin I-induced vasoconstriction was blocked. IW effectively inhibited aortic ACE activity ex vivo taken from SHRs after 14-weeks of oral treatment with IW. Furthermore, IW treated SHRs showed better endothelium-dependent vessel relaxation compared to placebo.This study shows strong ACE-inhibiting effects of IW, EW and WL in HUVECs and aorta. The peptides effectively counteract angiotensin-induced vasoconstriction and preserve endothelium-dependent vessel relaxation. Thus, tryptophan-containing peptides and particularly IW may serve as innovative food additives with the goal of protection from angiotensin II-induced worsening of vascular function.
Keywords: Angiotensin-converting enzyme; Arterial hypertension; Endothelial cells; Vessel tone; Tryptophan-containing peptides

MicroRNAs (miRNAs) are short, non-coding RNAs involved in almost all cellular processes. Epigallocatechin-3-O-gallate (EGCG) is a green tea polyphenol and is known to exert anti-arthritic effects by inhibiting genes associated with osteoarthritis (OA). This study was undertaken to investigate the global effect of EGCG on interleukin-1β (IL-1β)-induced expression of miRNAs in human chondrocytes.Human chondrocytes were derived from OA cartilage and then treated with EGCG and IL-1β. Human miRNA microarray technology was used to determine the expression profile of 1347 miRNAs. Microarray results were verified by taqman assays and transfection of chondrocytes with miRNA inhibitors.Out of 1347 miRNAs, EGCG up-regulated expression of 19 miRNAs and down-regulated expression of 17 miRNAs, whereas expression of 1311 miRNAs remains unchanged in IL-1β-stimulated human OA chondrocytes. Bioinformatics approach showed that 3`UTR of ADAMTS5 mRNA contains the ‘seed-matched-sequence’ for hsa-miR-140-3p. IL-1β-induced expression of ADAMTS5 correlated with down-regulation of hsa-miR-140-3p. Importantly, EGCG inhibited IL-1β-induced ADAMTS5 expression and up-regulated the expression of hsa-miR-140-3p. This EGCG-induced co-regulation between ADAMTS5 and hsa-miR-140-3p becomes reversed in OA chondrocytes transfected with anti-miR-140-3p.This study provides an important insight into the molecular basis of the reported anti-arthritic effects of EGCG. Our data indicate that the potential of EGCG in OA chondrocytes may be related to its ability to globally inhibit inflammatory response via modulation of miRNAs expressions.
Keywords: Osteoarthritis; MicroRNAs; Chondrocytes; Microarray; Hsa-miR-140-3p; ADAMTS5

Analysis of different innovative formulations of curcumin for improved relative oral bioavailability in human subjects by Martin Purpura; Ryan P. Lowery; Jacob M. Wilson; Haider Mannan; Gerald Münch; Valentina Razmovski-Naumovski (929-938).
The optimal health benefits of curcumin are limited by its low solubility in water and corresponding poor intestinal absorption. Cyclodextrins (CD) can form inclusion complexes on a molecular basis with lipophilic compounds, thereby improving aqueous solubility, dispersibility, and absorption. In this study, we investigated the bioavailability of a new γ-cyclodextrin curcumin formulation (CW8). This formulation was compared to a standardized unformulated curcumin extract (StdC) and two commercially available formulations with purported increased bioavailability: a curcumin phytosome formulation (CSL) and a formulation of curcumin with essential oils of turmeric extracted from the rhizome (CEO).Twelve healthy human volunteers participated in a double-blinded, cross-over study. The plasma concentrations of the individual curcuminoids that are present in turmeric (namely curcumin, demethoxycurcumin, and bisdemethoxycurcumin) were determined at baseline and at various intervals after oral administration over a 12-h period.CW8 showed the highest plasma concentrations of curcumin, demethoxycurcumin, and total curcuminoids, whereas CSL administration resulted in the highest levels of bisdemethoxycurcumin. CW8 (39-fold) showed significantly increased relative bioavailability of total curcuminoids (AUC0−12) in comparison with the unformulated StdC.The data presented suggest that γ-cyclodextrin curcumin formulation (CW8) significantly improves the absorption of curcuminoids in healthy humans.
Keywords: Curcumin; Cyclodextrin; Bioavailability; Humans; Plasma pharmacokinetics

Association between pre-pregnancy consumption of meat, iron intake, and the risk of gestational diabetes: the SUN project by Amelia Marí-Sanchis; Ginette Díaz-Jurado; F. Javier Basterra-Gortari; Carmen de la Fuente-Arrillaga; Miguel A. Martínez-González; Maira Bes-Rastrollo (939-949).
We assessed the association of total meat, processed, and unprocessed red meat and iron intake with the risk of developing gestational diabetes mellitus (GDM) in pregnant women.We conducted a prospective study among 3298 disease-free Spanish women participants of the SUN cohort who reported at least one pregnancy between December 1999 and March 2012. Meat consumption and iron intake were assessed at baseline through a validated, 136-item semi-quantitative, food frequency questionnaire. We categorized total, red, and processed meat consumption and iron intake into quartiles. Logistic regression models were used to adjust for potential confounders.We identified 172 incident cases of GDM. In the fully adjusted analysis, total meat consumption was significantly associated with a higher risk of GDM [OR = 1.67 (95% CI 1.06–2.63, p-trend 0.010)] for the highest versus the lowest quartile of consumption. The observed associations were particularly strong for red meat consumption [OR = 2.37 (95% CI 1.49–3.78, p-trend < 0.001)] and processed meat consumption [OR = 2.01 (95% CI 1.26–3.21, p-trend 0.003)]. Heme iron intake was also directly associated with GDM [OR = 2.21 (95% CI 1.37–3.58, p-trend 0.003)], although the association was attenuated and lost its statistical significance when we adjusted for red meat consumption [OR = 1.57 (95% CI 0.91–2.70, p-trend 0.213)]. No association was observed for non-heme and total iron intake, including supplements.Our overall findings suggest that higher pre-pregnancy consumption of total meat, especially red and processed meat, and heme iron intake, are significantly associated with an increased GDM risk in a Mediterranean cohort of university graduates.
Keywords: Gestational diabetes mellitus; Total meat; Red and processed meat; Heme iron intake; Mediterranean population

Mediation of psychosocial determinants in the relation between socio-economic status and adolescents’ diet quality by Nathalie Michels; Lisa Vynckier; Luis A. Moreno; Laurent Beghin; Alex de la O; Maria Forsner; Marcela Gonzalez-Gross; Inge Huybrechts; Isabel Iguacel; Antonio Kafatos; Mathilde Kersting; Catherine Leclercq; Yannis Manios; Ascension Marcos; Denes Molnar; Michael Sjöström; Kurt Widhalm; Stefaan De Henauw (951-963).
To examine the underlying reasons for the positive relation between socio-economic status (SES) and the diet quality of adolescents.In 2081 adolescents (12.5–17.5 years) of the European HELENA study, a continuous variable on diet quality via 2-day 24-h recalls was available. SES was reflected by parental education, parental occupation and family affluence. Mediation by several psychosocial determinants was tested: self-efficacy, availability at school and home, social support, barriers, benefits, awareness and some self-reported influencers (parents, school, taste, health, friends, food readily available, easy preparation, hunger, price and habits). Multiple mediation analyses were adjusted for age, sex and country.The availability of soft drinks and fruit at home, social support, parental influence, barriers, price influence, taste influence, health influence and food being readily available were significant mediators. The multiple mediation indirect effect accounted for 23–64% of the total effect. Both occupation and education and both maternal and paternal factors could be explained by the mediation. The unavailability of soft drinks was the strongest mediator (17–44% of the total effect).Up to 64% of the positive relation between SES and the diet quality in adolescence could be explained by several healthy eating determinants. Focusing on these factors in low-SES populations can minimize social inequalities in diet and health by improving the diet of these specific adolescents.
Keywords: Socio-economic class; Adolescents; Diet quality; Soft drink availability

Increasing secretion and production of glucagon-like peptide-1 (GLP-1) by continuous ingestion of certain food components has been expected to prevent glucose intolerance and obesity. In this study, we examined whether a physiological dose (5% weight in diet) of digestion-resistant maltodextrin (RMD) has a GLP-1-promoting effect in rats fed a high-fat and high-sucrose (HFS) diet.Rats were fed a control diet or the HFS (30% fat, 40% sucrose wt/wt) diet supplemented with 5% RMD or fructooligosaccharides (FOS) for 8 weeks or for 8 days in separated experiments. Glucose tolerance, energy intake, plasma and tissue GLP-1 concentrations, and cecal short-chain fatty acids concentrations were assessed.After 4 weeks of feeding, HFS-fed rats had significantly higher glycemic response to oral glucose than control rats, but rats fed HFS + RMD/FOS did not (approx. 50% reduction vs HFS rats). HFS + RMD/FOS-fed rats had higher GLP-1 responses (~twofold) to oral glucose, than control rats. After 8 weeks, visceral adipose tissue weight was significantly higher in HFS-fed rats than control rats, while HFS + RMD/FOS rats had a trend of reduced gain (~50%) of the tissue weight. GLP-1 contents and luminal propionate concentrations in the large intestine increased (>twofold) by adding RMD/FOS to HFS. Eight days feeding of RMD/FOS-supplemented diets reduced energy intake (~10%) and enhanced cecal GLP-1 production (~twofold), compared to HFS diet.The physiological dose of a prebiotic fiber promptly (within 8 days) promotes GLP-1 production in rats fed an obesogenic diet, which would help to prevent excess energy intake and fat accumulation.
Keywords: Resistant maltodextrin; Fructooligosaccharides; Glucagon-like peptide-1; High-fat and high-sucrose diet; Appetite; Adiposity

Ginger extract adjuvant to doxorubicin in mammary carcinoma: study of some molecular mechanisms by Nahla E. El-Ashmawy; Naglaa F. Khedr; Hoda A. El-Bahrawy; Hend E. Abo Mansour (981-989).
The present study aimed to investigate the molecular mechanisms underlying the anticancer properties of ginger extract (GE) in mice bearing solid Ehrlich carcinoma (SEC) and to evaluate the use of GE in combination with doxorubicin (DOX) as a complementary therapy against SEC.SEC was induced in 60 female mice. Mice were divided into four equal groups: SEC, GE, DOX and GE + DOX. GE (100 mg/kg orally day after day) and DOX (4 mg/kg i.p. for 4 cycles every 5 days) were given to mice starting on day 12 of inoculation. On the 28th day, blood samples were collected, mice were scarified, tumor volume was measured, and tumor tissues were excised.The anti-cancer effect of GE was mediated by activation of adenosine monophosphate protein kinase (AMPK) and down-regulation of cyclin D1 gene expression. GE also showed pro-apoptotic properties as evidenced by elevation of the P53 and suppression of nuclear factor-kappa B (NF-κB) content in tumor tissue. Co-administration of GE alongside DOX markedly increased survival rate, decreased tumor volume, and increased the level of phosphorylated AMPK (PAMPK) and improved related pathways compared to DOX group. In addition, the histopathological results demonstrated enhanced apoptosis and absence of multinucleated cells in tumor tissue of GE + DOX group.AMPK pathway and cyclin D1 gene expression could be a molecular therapeutic target for the anticancer effect of GE in mice bearing SEC. Combining GE and DOX revealed a greater efficacy as anticancer therapeutic regimen.
Keywords: AMPK; Cyclin D1; Doxorubicin; Ginger; Mammary carcinoma; Solid Ehrlich carcinoma

The study aimed to determine the effects of maternal low-protein (LP) diet on subcutaneous fat deposition of weaning piglets and the potential mechanism.Sows were fed either a standard protein (SP, 15 and 18% crude protein) or a LP diet (50% protein levels of SP) throughout pregnancy and lactation. Subcutaneous fat and blood were sampled from male piglets at 28 days of age. Serum biochemical metabolites and hormone concentrations were detected with the enzymatic colorimetric methods. Serum-free amino acid (FAA) levels were measured by amino acid auto-analyzer. The mRNA and protein were measured by qRT-PCR and Western blot.Body weight, back fat thickness, triglycerides concentrations in subcutaneous fat tissue, and serum, as well as FFA concentrations were significantly reduced in LP piglets when compared with SP piglets. Further studies showed that mRNA and protein expression of acetyl-CoA carboxylase and fatty acid synthetase, two key enzymes of de novo lipogenesis, were significantly reduced in LP piglets, while mRNA expression and the lipolytic enzymes activities of lipolysis genes, adipose triglyceride lipase and hormone-sensitive lipase, were significantly increased. Furthermore, expression of autophagy-related gene 7 and autophagy maker gene microtubule-associated protein 1A/1B-light chain 3 (LC 3) as well as the conversion of LC3I to LC3II were significantly elevated, along with the expression of activating transcription factor-4 and eukaryotic translation initiation factor-2a.These results indicate that amino acid starvation-induced autophagy is involved in reduced subcutaneous fat deposition in maternal LP weaning piglets, demonstrating links between maternal protein restriction and offspring fat deposition.
Keywords: Low-protein diet; Fat deposition; Activating transcription factor 4; Autophagy; Piglets

Erythrocyte fatty acid composition of Nepal breast-fed infants by Sigrun Henjum; Øyvind Lie; Manjeswori Ulak; Andrew L. Thorne-Lyman; Ram K. Chandyo; Prakash S. Shrestha; Wafaie W. Fawzi; Tor A. Strand; Marian Kjellevold (1003-1013).
Essential fatty acids play a critical role in the growth and development of infants, but little is known about the fatty acid status of populations in low-income countries. The objective was to describe the fatty acid composition of red blood cells (RBC) in breastfeed Nepali infants and a subsample of their mothers and to identify the main sources of fatty acids in the mother’s diet, as well as the fatty acid composition of breast milk.RBC fatty acid composition was analyzed in a random sample of 303 infants and 72 mother, along with 68 breastmilk samples. Fatty acid profiles of the most important dietary fat sources were analyzed. Information on mother’s diet and intake of fat was collected by three 24-h dietary recalls.In infant RBC’s, docosahexaenoic acid (DHA) was the main n-3 fatty acid, and arachidonic acid (AA) was the major n-6 fatty acid. Total n-6 PUFA was three times higher than total n-3 PUFA. Height-for-age (HAZ) was positively associated with DHA status and AA status in multivariable models. The concentration of all fatty acids was higher in children, compared to mothers, except Total n-6 PUFA and Linoleic acid (LA) where no differences were found. The mother’s energy intake from fat was 13% and cooking oil (sesame, mustard, soybean or sunflower oil) contributed 52% of the fat intake.RBC-DHA levels in both infants and mother was unexpected high taking into account few dietary DHA sources and the low DHA concentrations in breastmilk.
Keywords: Polyunsaturated fatty acids; Plasma phospholipids; DHA; AA; Breast-fed children; Breastmilk

Calcium intake in winter pregnancy attenuates impact of vitamin D inadequacy on urine NTX, a marker of bone resorption by Eileen C. O’Brien; Mark T. Kilbane; Malachi J. McKenna; Ricardo Segurado; Aisling A. Geraghty; Fionnuala M. McAuliffe (1015-1023).
Pregnancy is characterised by increased bone turnover, but high bone turnover with resorption exceeding formation may lead to negative maternal bone remodelling. Recent studies are conflicting regarding the effect of calcium on skeletal health in pregnancy. The aim of this study was to examine the seasonal effect of serum 25-hydroxyvitamin D (25OHD) and dietary calcium on a marker of bone resorption.This was prospective study of 205 pregnant women [two cohorts; early pregnancy at 13 weeks (n = 96), and late pregnancy at 28 weeks (n = 109)]. Serum 25OHD and urine cross-linked N-telopeptides of type I collagen (uNTX) were measured at both time points. Intakes of vitamin D and calcium were recorded using 3-day food diaries at each trimester.Compared to summer pregnancies, winter pregnancies had significantly lower 25OHD and significantly higher uNTX. Higher calcium intakes were negatively correlated with uNTX in winter, but not summer. In late pregnancy, compared to those reporting calcium intakes ≥1000 mg/day, intakes of <1000 mg/day were associated with a greater increase in uNTX in winter pregnancies than in summer (41.8 vs. 0.9%). Increasing calcium intake in winter by 200 mg/day predicted a 13.3% reduction in late pregnancy uNTX.In late pregnancy, during winter months when 25OHD is inadequate, intakes of dietary calcium <1000 mg/day were associated with significantly increased bone resorption (uNTX). Additional dietary calcium is associated with reduced bone resorption in late pregnancy, with greater effect observed in winter. Further research regarding optimal dietary calcium and 25OHD in pregnancy is required, particularly for women gestating through winter.
Keywords: Pregnancy; Calcium; 25OHD; Vitamin D; Bone turnover; Season

Chlorella vulgaris (CV) has exhibited immune-enhancing and protective activities against cancer and infections. However, there is an increasing concern about the use of Chlorella species in human, regarding its various molecules with antigenic features found in infectious microorganisms. Our goal was to investigate the impact of higher concentrations of CV on tumor growth in spontaneous mouse mammary tumor (SMMT) models.Balb/c mice were daily given CV powder at doses of 0, 200, or 300 mg/kg for 42 days (CONTROL, CV200, and CV300 groups, respectively; n = 6/group). On day 14, the SMMT was inoculated. Tumor volume (TV) and body weight (BW) were monitored on 5-day intervals following tumor challenge. On day 43, blood, spleen, lungs, and tumor tissues were collected. Histopathological examinations on lungs and tumor tissues were performed following hematoxylin–eosin staining. Intratumor expression of 27 genes was assessed by real-time PCR. Total IgG, IFNγ, and IL-4 levels in serum and spleen culture supernatant were measured by ELISA.The TV/BW index showed significant increase in the CV200 group compared to the CONTROL (p = 0.047). The CV200 tumors exhibited more malignant phenotype, higher angiogenesis rate, and lower peritumoral neutrophil and macrophage-to-lymphocyte infiltration ratio compared to the CONTROL. Serum concentrations of IFNγ, IL-4, and IgG were declined, and the spleen IFNγ and IgG production was higher in the CV200 compared to the CONTROL. The IL-1β, IL-10, TGFβ1, FOXP3, HO-1, Gr1, CD11b, PCNA, LCN2, iNOS2, VEGFR2, CD31, and CD105L expressions were markedly increased in the CV200 tumors compared to the CONTROL (p = 0.001, 0.002, 0.006, 0.021, 0.004, 0.030, 0.016, 0.031, 0.025, 0.008, 0.014, 0.022, and 0.037, respectively). The changes in cytokine, IgG and gene expression values considerably correlated with tumor size, as well as with each other.Our data provided evidence that C. vulgaris at a specific dose (200 mg/kg) promoted tumor growth in a mammary tumor model. This consequence might reflect an immune derangement in favor of developing a protumor microenvironment. However, this hypothesis needs to be further investigated in future.
Keywords: Angiogenesis; Chlorella vulgaris ; Immunosuppression; Spontaneous mouse mammary tumor; Tumor progression

Comparison of meal patterns across five European countries using standardized 24-h recall (GloboDiet) data from the EFCOVAL project by Min Kyung Park; Heinz Freisling; Ena Huseinovic; Anna Winkvist; Inge Huybrechts; Sandra Patricia Crispim; Jeanne H. M. de Vries; Anouk Geelen; Maryse Niekerk; Caroline van Rossum; Nadia Slimani (1045-1057).
To examine meal patterns in terms of frequency and circadian timing of eating in five European countries participating in the EFCOVAL project.In this cross-sectional study, 559 men and women, aged 44–65 years, were recruited in Belgium, the Czech Republic, France (Southern part), The Netherlands, and Norway. Dietary data were collected by trained interviewers using standardized computerised 24-h recalls (GloboDiet). Means ± SE of (1) eating frequency, (2) overnight fasting, and (3) time between eating occasions were estimated by country using means from 2 days of 24-h recalls. We also estimated the frequency of eating occasions per hour by country as well as the proportional energy intake of meals/snacks by country compared to the mean energy intake of all countries.Mean eating frequency ranged from 4.3 times/day in France to 7.1 times/day in The Netherlands (p < 0.05). Mean overnight fasting was shortest in the Netherlands (9.2 h) and longest in Czech Republic (10.9 h) (p < 0.05). Mean time between single eating occasions was shortest in The Netherlands (2.4 h) and longest in France (4.3 h) (p < 0.05). Different patterns of energy intake by meals and snacks throughout the day were observed across the five countries.We observed distinct differences in meal patterns across the five European countries included in the current study in terms of frequency and circadian timing of eating, and the proportion of energy intake from eating occasions.
Keywords: Meal/snack pattern; Snacking; Eating frequency; EFCOVAL; GloboDiet/EPIC-Soft; Food consumption

Changes in plasma concentrations of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D during pregnancy: a Brazilian cohort by Amanda C. Cunha Figueiredo; Paula Guedes Cocate; Amanda R. Amorim Adegboye; Ana Beatriz Franco-Sena; Dayana R. Farias; Maria Beatriz Trindade de Castro; Alex Brito; Lindsay H. Allen; Rana R. Mokhtar; Michael F. Holick; Gilberto Kac (1059-1072).
To characterize the physiological changes in 25-hydroxyvitamin D [25(OH)D] and 1,25-dihydroxyvitamin D [1,25(OH)2D] throughout pregnancy.Prospective cohort of 229 apparently healthy pregnant women followed at 5th–13th, 20th–26th, and 30th–36th gestational weeks. 25(OH)D and 1,25(OH)2D concentrations were measured by LC–MS/MS. Statistical analyses included longitudinal linear mixed-effects models adjusted for parity, season, education, self-reported skin color, and pre-pregnancy BMI. Vitamin D status was defined based on 25(OH)D concentrations according to the Endocrine Society Practice Guideline and Institute of Medicine (IOM) for adults.The prevalence of 25(OH)D <75 nmol/L was 70.4, 41.0, and 33.9%; the prevalence of 25(OH)D <50 nmol/L was 16.1, 11.2, and 10.2%; and the prevalence of 25(OH)D <30 nmol/L was 2, 0, and 0.6%, at the first, second, and third trimesters, respectively. Unadjusted analysis showed an increase in 25(OH)D (β = 0.869; 95% CI 0.723–1.014; P < 0.001) and 1,25(OH)2D (β = 3.878; 95% CI 3.136–4.620; P < 0.001) throughout pregnancy. Multiple adjusted analyses showed that women who started the study in winter (P < 0.001), spring (P < 0.001), or autumn (P = 0.028) presented a longitudinal increase in 25(OH)D concentrations, while women that started during summer did not. Increase of 1,25(OH)2D concentrations over time in women with insufficient vitamin D (50–75 nmol/L) at baseline was higher compared to women with sufficient vitamin D (≥75 nmol/L) (P = 0.006).The prevalence of vitamin D inadequacy varied significantly according to the adopted criteria. There was a seasonal variation of 25(OH)D during pregnancy. The women with insufficient vitamin D status present greater longitudinal increases in the concentrations of 1,25(OH)2D in comparison to women with sufficiency.
Keywords: Vitamin D; Pregnancy; Micronutrients; Cohort; Tropical country; Seasons

Effect of AHA dietary counselling on added sugar intake among participants with metabolic syndrome by Lijuan Zhang; Sherry Pagoto; Christine May; Barbara Olendzki; Katherine L.Tucker; Carolina Ruiz; Yu Cao; Yunsheng Ma (1073-1082).
High added sugar consumption has been associated with the development of metabolic syndrome (MetS). The American Heart Association (AHA) diet is designed to prevent and treat MetS; however, it remains unclear whether the AHA diet is effective on decreasing added sugar consumption. The aim of our study was to evaluate the effect of the AHA dietary counselling on added sugar consumption among participants with MetS.The AHA dietary counselling was conducted among 119 participants with MetS from June 2009 to January 2014 ( NCT00911885). Unannounced 24-hour recalls were collected at baseline, 3, 6 and 12 months. Added sugar consumption patterns over time were examined using linear mixed models.After 1-year dietary counselling, intake of added sugars decreased by 23.8 g/day (95% CI 15.1, 32.4 g/day); intake of nonalcoholic beverages dropped from the leading contributor of added sugar intake to number 7 (from 11.9 to 4.4%); the Alternative Healthy Eating Index (AHEI) score increased by 5.4 (95% CI 2.9, 8.0); however, added sugar intake for 48% participants still exceeded the recommendation. Added sugar intake per meal among different meal type was similar (24.2–25.8%) at baseline. After the 1-year dietary counselling, breakfast became the major resource of added sugar intake (33.3%); the proportion of added sugar intake from snacks decreased from 25.8% (CI 23.1, 28.5%) to 20.9% (CI 19.6, 22.3%).Although the consumption of added sugars in participants with MetS decreased after the 1-year AHA dietary counselling, added sugar intake from majority of participants still exceeds recommended limits. Actions of successful public health strategies that focus on reducing added sugar intake are needed.
Keywords: Added sugar; Metabolic syndrome; AHA dietary counselling

Effect of administration of high-protein diet in rats submitted to resistance training by Thiago da Rosa Lima; Eudes Thiago Pereira Ávila; Géssica Alves Fraga; Mariana de Souza Sena; Arlyson Batista de Souza Dias; Paula Caroline de Almeida; Joice Cristina dos Santos Trombeta; Roberto Carlos Vieira Junior; Amílcar Sabino Damazo; James Wilfred Navalta; Jonato Prestes; Fabrício Azevedo Voltarelli (1083-1096).
Although there is limited evidence regarding the pathophysiological effects of a high-protein diet (HD), it is believed that this type of diet could overload the body and cause damage to the organs directly involved with protein metabolism and excretion. The aim of this study was to verify the effects of HD on biochemical and morphological parameters of rats that completed a resistance training protocol (RT; aquatic jump) for 8 weeks.Thirty-two adult male Wistar rats were divided into four groups (n = 8 for each group): sedentary normal protein diet (SN-14%), sedentary high-protein diet (SH-35%), trained normal protein diet (TN-14%), and trained high-protein diet (TH-35%). Biochemical, tissue, and morphological measurements were made.Kidney (1.91 ± 0.34) and liver weights (12.88 ± 1.42) were higher in the SH. Soleus muscle weight was higher in the SH (0.22 ± 0.03) when compared to all groups. Blood glucose (123.2 ± 1.8), triglycerides (128.5 ± 44.0), and HDL cholesterol levels (65.7 ± 20.9) were also higher in the SH compared with the other experimental groups. Exercise reduced urea levels in the trained groups TN and TH (31.0 ± 4.1 and 36.8 ± 6.6), respectively. Creatinine levels were lower in TH and SH groups (0.68 ± 0.12; 0.54 ± 0.19), respectively. HD negatively altered renal morphology in SH, but when associated with RT, the apparent damage was partially reversed. In addition, the aquatic jump protocol reversed the damage to the gastrocnemius muscle caused by the HD.A high-protein diet promoted negative metabolic and morphological changes, while RT was effective in reversing these deleterious effects.
Keywords: Dietary management; High-protein diet; Resistance training; Aquatic jump training; Tissue morphology

The satiating effect of animal vs plant proteins remains unknown. The present study examined the effects of breakfasts containing animal proteins [milk (AP)], a blend of plant proteins [oat, pea and potato (VP)] or 50:50 mixture of the two (MP) compared with a carbohydrate-rich meal (CHO) on appetite, energy intake (EI) and metabolic measures.A total of 28 males [mean age 27.4 (±SD 4.2) years, BMI 23.4 (±2.1) kg/m2] consumed three isoenergetic (1674 kJ) rice puddings matched for energy density and macronutrient content as breakfast (25% E from protein) in a single-blind, randomised, cross over design. Appetite ratings and blood samples were collected and assessed at baseline and every 30 and 60 min, respectively, until an ad libitum test meal was served 3.5 h later. Free-living appetite was recorded hourly and EI in weighed food records for the remainder of the day.No differences in subjective appetite ratings were observed after consumption of the AP, VP and MP. Furthermore, there were no differences between the AP, VP, MP and CHO breakfasts in ad libitum EI and self-reported EI during the remainder of the day. Although insulin metabolism was not affected, CHO induced a higher glucose response (P = 0.001) and total amino acids concentration was in the order of AP = MP > VP > CHO breakfast (P = 0.001).Manipulating the protein source of foods consumed as breakfast, elicited comparable effects on appetite and EI at both laboratory and free-living environment in healthy men.
Keywords: Milk proteins; Pea protein; Appetite; Energy intake; Amino acids; Insulin

Starch-enriched diet modulates the glucidic profile in the rat colonic mucosa by Maria Gabriella Gabrielli; Daniele Tomassoni (1109-1121).
The protective function of the intestinal mucosa largely depends on carbohydrate moieties that as a part of glycoproteins and glycolipids form the epithelial glycocalyx or are secreted as mucins. Modifications of their expression can be induced by an altered intestinal microenvironment and have been associated with inflammatory disorders and colorectal cancer. Given the influence of dietary factors on the gut ecosystem, here we have investigated whether a long term feeding on a starch-rich diet can modulate the glucidic profile in the colonic mucosa of rats.Animals were divided into two groups and maintained for 9 months at different diets: one group was fed a standard diet, the second was fed a starch-enriched diet. Samples of colonic mucosa, divided in proximal and distal portions, were processed for microscopic analysis. Conventional stainings and lectin histochemistry were applied to identify acidic glycoconjugates and specific sugar residues in oligosaccharide chains, respectively. Some lectins were applied on adjacent sections after sialidase/fucosidase digestion, deacetylation, and oxidation to characterize either terminal dimers or sialic acid acetylation.An increase in sulfomucins was found to be associated with the starch-enriched diet that affected also the expression of several sugar residues as well as fucosylated and sialylated sequences in both proximal and distal colon.Although the mechanisms leading to such a modulation are at present unknown, either an altered intestinal microbiota or a dysregulation of glycosylation patterns might be responsible for the types and distribution of changes in the glucidic profile here observed.
Keywords: Lectin histochemistry; Colon; Sialic acid; Starch-rich diet; Glycosylation

Immune dysfunction and increased oxidative stress state in diet-induced obese mice are reverted by nutritional supplementation with monounsaturated and n-3 polyunsaturated fatty acids by Caroline Hunsche; Oskarina Hernandez; Alina Gheorghe; Ligia Esperanza Díaz; Ascensión Marcos; Mónica De la Fuente (1123-1135).
Obesity is associated with impaired immune defences and chronic low levels of inflammation and oxidation. In addition, this condition may lead to premature aging. The aim of the study was to evaluate the effects of a nutritional supplementation with monounsaturated and n-3 polyunsaturated fatty acids on several functions and oxidative stress parameters in peritoneal immune cells of obese mice, as well as on the life span of these animals.Obesity was induced in adult female ICR/CD1 by the administration of a high-fat diet (HFD) for 14 weeks. During the last 6 weeks of HFD feeding, one group of obese mice received the same HFD, supplemented with 1500 mg of 2-hydroxyoleic acid (2-OHOA) and another with 3000 mg of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Several functions and oxidative stress parameters of peritoneal leukocytes were evaluated.The groups of obese mice treated with 2-OHOA or with EPA and DHA showed a significant improvement in several functions such as chemotaxis, phagocytosis, digestion capacity, Natural killer activity and lymphoproliferation in response to mitogens. All of these functions, which were decreased in obese mice, increased reaching similar levels to those found in non-obese controls. Both treatments also improved oxidative stress parameters such as xanthine oxidase activity, which decreased, catalase activity and glutathione levels, which increased.These data suggest that dietary supplementation with monounsaturated and n-3 polyunsaturated fatty acids could be an effective nutritional intervention to restore the immune response and oxidative stress state, which are impaired in obese mice.
Keywords: Obese mice; Monounsaturated fatty acids; n-3 Polyunsaturated fatty acids; Immune function; Oxidative stress

Sesamin extends lifespan through pathways related to dietary restriction in Caenorhabditis elegans by Yumiko Nakatani; Yukie Yaguchi; Tomomi Komura; Masakazu Nakadai; Kenji Terao; Eriko Kage-Nakadai; Yoshikazu Nishikawa (1137-1146).
Sesamin, a polyphenolic compound found in sesame seeds, has been reported to exert a variety of beneficial health effects. We have previously reported that sesamin increases the lifespan of Caenorhabditis elegans. In this study, we investigated the molecular mechanisms underlying the longevity effect of sesamin in C. elegans.Starting from three days of age, Caenorhabditis elegans animals were fed a standard diet alone or supplemented with sesamin. A C. elegans genome array was used to perform a comprehensive expression analysis. Genes that showed differential expression were validated using real-time PCR. Mutant or RNAi-treated animals were fed sesamin, and the lifespan was determined to identify the genes involved in the longevity effects of sesamin.The microarray analysis revealed that endoplasmic reticulum unfolded protein response-related genes, which have been reported to show decreased expression under conditions of SIR-2.1/Sirtuin 1 (SIRT1) overexpression, were downregulated in animals supplemented with sesamin. Sesamin failed to extend the lifespan of sir-2.1 knockdown animals and of sir-2.1 loss-of-function mutants. Sesamin was also ineffective in bec-1 RNAi-treated animals; bec-1 is a key regulator of autophagy, and is necessary for longevity induced by sir-2.1 overexpression. Furthermore, the heterozygotic mutation of daf-15, which encodes the target of rapamycin (TOR)-binding partner Raptor, abolished lifespan extension by sesamin. Moreover, sesamin did not prolong the lifespan of loss-of-function mutants of aak-2, which encodes the AMP-activated protein kinase (AMPK).Sesamin extends the lifespan of C. elegans through several dietary restriction-related signaling pathways, including processes requiring SIRT1, TOR, and AMPK.
Keywords: Sesamin; Lifespan; Caenorhabditis elegans ; Dietary restriction

Prevalence of childhood hypertension and hypertension phenotypes by weight status and waist circumference: the Healthy Growth Study by Yannis Manios; K. Karatzi; A. D. Protogerou; G. Moschonis; C. Tsirimiagou; O. Androutsos; C. Lionis; G. P. Chrousos (1147-1155).
The aim of the present study was to report for the first time the prevalence of hypertension and its phenotypes in obese children and in children with central obesity in a large sample of Greek children.A regionally representative sample of 2263 schoolchildren (50.3% boys) (9–13 years) having full data on blood pressure assessment, physical examination, anthropometric, and physical activity participated in a cross-sectional study in Greece.Prevalence of stage 1 and 2 hypertension, of isolated systolic hypertension (ISH) and of combined systolic or diastolic hypertension, was significantly higher for obese children and children on the 3rd tertile of waist circumference in the total sample, as well as in each gender separately. ISH was the most prevalent phenotype reaching 24.3% in obese children and 17.5% in children on the highest tertile of waist circumference. Obese children and children on the highest tertile of waist circumference had 6.31 times and 3.94 times, respectively, higher likelihood to have abnormal systolic or diastolic blood pressure (SBP or DBP) than their normal-weight counterparts.Prevalence of hypertension and especially ISH in obese children and in children with central obesity in Greece are among the highest reported in Europe. Future public health initiatives should aim to prevent or tackle several underlying factors related to childhood hypertension, focusing primarily on children with excess body weight.
Keywords: Childhood hypertension; Adolescent hypertension; Overweight; Childhood obesity; Central obesity

The α’ subunit of β-conglycinin and various glycinin subunits of soy are not required to modulate hepatic lipid metabolism in rats by Cynthia Chatterjee; Jiajie Liu; Carla Wood; Christine Gagnon; Elroy R. Cober; Judith A. Frégeau-Reid; Stephen Gleddie; Chao-Wu Xiao (1157-1168).
This study examined the effect of soy proteins with depletion of different subunits of the two major storage proteins, β-conglycinin and glycinin, on hepatic lipids and proteins involved in lipid metabolism in rats, since the bioactive component of soy responsible for lipid-lowering is unclear.Weanling Sprague Dawley rats were fed diets containing either 20% casein protein in the absence (casein) or presence (casein + ISF) of isoflavones or 20% alcohol-washed soy protein isolate (SPI) or 20% soy protein concentrates derived from a conventional (Haro) or 2 soybean lines lacking the α’ subunit of β-conglycinin and the A1-3 (1TF) or A1-5 (1a) subunits of glycinin. After 8 weeks, the rats were necropsied and liver proteins and lipids were extracted and analysed.The results showed that soy protein diets reduced lipid droplet accumulation and content in the liver compared to casein diets. The soy protein diets also decreased the level of hepatic mature SREBP-1 and FAS in males, with significant decreases in diets 1TF and 1a compared to the casein diets. The effect of the soy protein diets on female hepatic mature SREBP-1, FAS, and HMGCR was confounded since casein + ISF decreased these levels compared to casein alone perhaps muting the decrease by soy protein. A reduction in both phosphorylated and total STAT3 in female livers by ISF may account for the gender difference in mechanism in the regulation and protein expression of the lipid modulators.Overall, soy protein deficient in the α’ subunit of β-conglycinin and A1-5 subunits of glycinin maintain similar hypolipidemic function compared to the conventional soy protein. The exact bioactive component(s) warrant identification.
Keywords: Soy protein; β-Conglycinin; Glycinin; Liver; Lipid metabolism; Metabolic disease; Rats

Dietary blueberry improves cognition among older adults in a randomized, double-blind, placebo-controlled trial by Marshall G. Miller; Derek A. Hamilton; James A. Joseph; Barbara Shukitt-Hale (1169-1180).
As populations shift to include a larger proportion of older adults, the necessity of research targeting older populations is becoming increasingly apparent. Dietary interventions with blueberry have been associated with positive outcomes in cell and rodent models of aging. We hypothesized that dietary blueberry would improve mobility and cognition among older adults.In this study, 13 men and 24 women, between the ages of 60 and 75 years, were recruited into a randomized, double-blind, placebo-controlled trial in which they consumed either freeze-dried blueberry (24 g/day, equivalent to 1 cup of fresh blueberries) or a blueberry placebo for 90 days. Participants completed a battery of balance, gait, and cognitive tests at baseline and again at 45 and 90 days of intervention.Significant supplement group by study visit interactions were observed on tests of executive function. Participants in the blueberry group showed significantly fewer repetition errors in the California Verbal Learning test (p = 0.031, ηp 2 = 0.126) and reduced switch cost on a task-switching test (p = 0.033, ηp 2 = 0.09) across study visits, relative to controls. However, no improvement in gait or balance was observed.These findings show that the addition of easily achievable quantities of blueberry to the diets of older adults can improve some aspects of cognition.
Keywords: Aging; Blueberry; Cognition; Gait; Postural Sway

A low-dose, 6-week bovine colostrum supplementation maintains performance and attenuates inflammatory indices following a Loughborough Intermittent Shuttle Test in soccer players by Yiannis Kotsis; Anastasia Mikellidi; Cleopatra Aresti; Eleni Persia; Aristomenis Sotiropoulos; Demosthenes B. Panagiotakos; Smaragdi Antonopoulou; Tzortzis Nomikos (1181-1195).
The aim of the study was to investigate the effect of a 6-week, low-dose bovine colostrum (BC) supplementation on exercise-induced muscle damage (EIMD) and performance decline in soccer players following the Loughborough Intermittent Shuttle Test (LIST) during a competitive season period.In a double-blind, randomized, placebo-controlled design, two groups of soccer players were allocated to a 3.2 g/day of whey protein (WP, N = 8) or BC (N = 10) and performed a pre- and a post-supplementation LIST. Maximum isometric voluntary contraction, squat jump (SQJ), countermovement jump, muscle soreness, blood cell counts, creatine kinase (CK), C-reactive protein (CRP) and interleukin-6 (IL-6) were monitored for 2, 24, 48, 72 h post-LIST.LIST induced transient increases in leukocytes, granulocytes, CK, muscle soreness, CRP, IL-6 and declines in lymphocytes and performance indices. Supplementation resulted in a faster recovery of SQJ, CK and CRP compared to pre-supplementation kinetics (trial × time: p = 0.001, 0.056, 0.014, respectively) and lower incremental area under the curve (iAUC) for IL-6, only in the BC group [pre-: 31.1 (6.78–46.9), post-: 14.0 (−0.16 to 23.5) pg h/ml, p = 0.034]. Direct comparison of the two groups after supplementation demonstrated higher iAUC of SQJ [WP: −195.2 (−229.0 to (−52.5)), BC: −15.8 (−93.2 to 16.8) cm h, p = 0.034], a trend for lower iAUC of CK in the BC group [WP: 18,785 (4651–41,357), BC: 8842 (4807–14,802) U h/L, p = 0.081] and a significant intervention × time interaction for CRP (p = 0.038) in favor of BC.Post-exercise EIMD may be reduced and performance better maintained by a low dose of BC administration following LIST in soccer players.
Keywords: Bovine colostrum; Exercise-induced muscle damage; Loughborough Intermittent Shuttle Test; Soccer; Inflammation; Squat jump

The relationship between carbohydrate quality and the prevalence of metabolic syndrome: challenges of glycemic index and glycemic load by Mariane de Mello Fontanelli; Cristiane Hermes Sales; Antonio Augusto Ferreira Carioca; Dirce Maria Marchioni; Regina Mara Fisberg (1197-1205).
To estimate the prevalence of metabolic syndrome (MetS) and its components in adults and older adults residents of São Paulo, the association of MetS with the glycemic index (GI) and glycemic load (GL) and the foods that contribute to dietary GI and GL in this population.Data from 591 adults and older adults participants in the Health Survey of São Paulo were used. This is a cross-sectional, population-based study with a complex multistage sample design of residents in the urban area of the municipality. Dietary consumption data, anthropometric measurements, blood pressure and blood samples were collected. The associations between GI, GL and MetS and its components were tested using logistic regression models, considering the sample design of the study.The prevalence of MetS in the adult and older adults residents of São Paulo was 30.3%. There was no association between GI, GL and MetS. GI and GL were positively associated with low high-density lipoprotein cholesterol (HDL-c), OR = 1.113 (95% CI 1.007–1.230) and OR = 1.019 (95% CI 1.002–1.037), respectively. GL was inversely associated with high blood pressure and this association differed by age group (OR = 0.981; 95% CI 0.964–0.998). Foods that most contributed to dietary GI and GL were sugar, white rice and French bread.Considering the high prevalence of low HDL-c in the population of São Paulo, GI and GL may contribute to the nutritional therapy of this dyslipidemia. However, findings should be treated with caution, considering several conflicting results between studies.
Keywords: Metabolic Syndrome; Glycemic Index; Glycemic Load; Cross-sectional studies; Prevalence

Dietary inflammatory index and risk of renal cancer in the Iowa Women’s Health Study by Nitin Shivappa; Cindy K. Blair; Anna E. Prizment; David R. Jacobs Jr.; James R. Hébert (1207-1213).
The association between inflammatory potential of diet and renal cancer risk has not been investigated.In this study, we explored the association between the dietary inflammatory index (DII) and risk of renal cancer in the Iowa Women’s Health Study. From 1986 to 2011, 33,817 women initially recruited at 55–69 years of age were followed for incident renal cancers (n = 263). The DII was computed based on dietary intake assessed using a reproducible and valid 121-item food frequency questionnaire. Cox proportional hazards regression was used to estimate hazard ratios (HR) adjusting for age, body mass index, energy intake, smoking status, education, pack years of smoking, hypertension, and hormone replacement therapy.Multivariable analyses revealed positive association between higher DII scores and renal cancer risk (HR for DIIcontinuous: 1.07 per unit increase in DII (corresponding to 10% change in the DII range in the current study); 95% CI 1.00, 1.15; HR for DIItertile3vs1 = 1.52; 95% CI 1.09, 2.13). Stratified analyses produced slightly stronger associations between DII and renal cancer risk among women with BMI <30 kg/m2 (HRTertile3vs1 = 1.57; 95% CI = 1.04, 2.36) and ever smokers (HRtertile3vs1 = 2.35; 95% CI = 1.22, 4.55), although the corresponding interaction p values were not significant.Pro-inflammatory diet, as indicated by higher DII scores, was associated with increased renal cancer risk.
Keywords: Dietary inflammatory index (DII); Diet; Inflammation; Renal cancer risk; Cohort

Attenuation of liver cancer development by oral glycerol supplementation in the rat by Alejo M. Capiglioni; Florencia Lorenzetti; Ariel D. Quiroga; Juan P. Parody; María T. Ronco; Gerardo B. Pisani; María C. Carrillo; María P. Ceballos; María de Luján Alvarez (1215-1224).
Glycerol usage is increasing in food industry for human and animal nutrition. This study analyzed the impact of glycerol metabolism when orally supplemented during the early stage of rat liver carcinogenesis.Wistar rats were subjected to a 2-phase model of hepatocarcinogenesis (initiated-promoted, IP group). IP animals also received glycerol by gavage (200 mg/kg body weight, IPGly group).Glycerol treatment reduced the volume of preneoplastic lesions by decreasing the proliferative status of liver foci, increasing the expression of p53 and p21 proteins and reducing the expression of cyclin D1 and cyclin-dependent kinase 1. Besides, apoptosis was enhanced in IPGly animals, given by an increment of Bax/Bcl-2 ratio, Bad and PUMA mitochondrial expression, a concomitant increase in cytochrome c release and caspase-3 activation. Furthermore, hepatic levels of glycerol phosphate and markers of oxidative stress were increased in IPGly rats. Oxidative stress intermediates act as intracellular messengers, inducing p53 activation and changes in JNK and Erk signaling pathways, with JNK activation and Erk inhibition.The present work provides novel data concerning the preventive actions of glycerol during the development of liver cancer and represents an economically feasible intervention to treat high-risk individuals.
Keywords: Proliferation; Apoptosis; Glycerol; Liver preneoplasia; Oxidative stress

Prospective association between adherence to the Mediterranean diet and risk of depressive symptoms in the French SU.VI.MAX cohort by Moufidath Adjibade; Karen E. Assmann; Valentina A. Andreeva; Cédric Lemogne; Serge Hercberg; Pilar Galan; Emmanuelle Kesse-Guyot (1225-1235).
This study examines whether adherence to the Mediterranean Diet (MD) measured by several dietary indexes was associated with incident depressive symptoms in a large French cohort.The study sample consisted of 3523 participants from the Supplémentation en Vitamines et Minéraux Antioxydants (SU.VI.MAX) cohort who had at least three dietary records at baseline during the first 2 years of follow-up (1994–1996), free of depression at the beginning of the study (1996–1997) and available Center for Epidemiologic Studies Depression Scale (CES-D) data at the end of follow-up (2007–2009). The rMED was computed. Incident depressive symptoms were defined by a CES-D score ≥17 for men and ≥23 for women in 2007–2009. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated using multivariable logistic regression models. Several sensitivity analyses were performed.In the present study, 172 incident cases of depressive symptoms were identified during the follow-up (mean = 12.6 years). After adjustment for a wide range of potential confounders, adherence to the rMED score (continuous variable) was significantly associated with incident depressive symptoms in men (OR 0.91; 95% CI 0.83–0.99; p = 0.03), but not in women. Use of the Literature-Based Adherence Score to the Mediterranean Diet (LAMD) and the classic MD score (MDS) provide similar findings.In the current study, higher adherence to the Mediterranean Diet at midlife was associated with a lower risk of incident depressive symptoms, particularly in men, increasing scientific evidence for a beneficial role of Mediterranean Diet on health. Further investigations in particular among women are needed.
Keywords: Mental health; Depressive symptoms; Mediterranean diet; Prospective study

Oxidative balance score and serum γ-glutamyltransferase level among Korean adults: a nationwide population-based study by A.-Ra Cho; Yu-Jin Kwon; Hyoung-Ji Lim; Hye Sun Lee; Sinae Kim; Jae-Yong Shim; Hye-Ree Lee; Yong-Jae Lee (1237-1244).
The oxidative balance score (OBS) comprises dietary and non-dietary lifestyle pro-oxidants and antioxidants. Elevated serum γ-glutamyltransferase (GGT) level has currently emerged as a biomarker of oxidative stress. In this study, we examined whether OBS was inversely associated with serum GGT level and whether OBS could be a useful marker to predict GGT among Korean adults.This cross-sectional study was based on data obtained from the 2010 and 2011 Korea National Health and Nutrition Examination Survey. 2087 men and 2071 women were included in final analysis. The OBS was divided into five equal interval categories, and GGT was dichotomized into low and high using its sex-specific median value. Multiple logistic regression analysis was conducted to assess the association between OBS categories and high GGT.Compared with the lowest OBS category as reference, the multivariable adjusted ORs (95% CIs) for the highest OBS category of men and women were 0.05 (0.01–0.19) and 0.27 (0.09–0.78), respectively (p for trend <0.01).A higher OBS that indicates a predominance of antioxidant over pro-oxidant exposure was strongly inversely associated with GGT level among Korean adults.
Keywords: Oxidative balance score; Oxidative stress; γ-Glutamyltransferase; Inflammation; Biomarker

Adherence to a healthy diet in relation to cardiovascular incidence and risk markers: evidence from the Caerphilly Prospective Study by Elly Mertens; Oonagh Markey; Johanna M. Geleijnse; Julie A. Lovegrove; D. Ian Givens (1245-1258).
Epidemiological findings indicate that higher adherence to a healthy diet may lower cardiovascular disease (CVD) risk. The present study aimed to investigate whether adherence to a healthy diet, assessed by the Healthy Diet Indicator (HDI), Dietary Approaches to Stop Hypertension (DASH) score, and Alternative Healthy Eating Index 2010 (AHEI-2010), was associated with CVD incidence and risk markers.Included in the present analyses were data from 1867 middle-aged men, aged 56.7 ± 4.5 years at baseline, recruited into the Caerphilly Prospective Study. Adherence to a healthy diet was examined in relation to CVD, coronary heart disease (CHD), and stroke incidence (Cox regression), and risk markers (linear regression) with adjustment for relevant confounders.The DASH score was inversely associated with CVD [hazard ratio (HR) 0.81; 95% confidence interval (CI) 0.66, 0.99], and stroke (HR 0.61; 95% CI 0.42, 0.88) incidence, but not with CHD after an average of 16.6 year follow-up, and with diastolic blood pressure, after 12 year follow-up. The AHEI-2010 was inversely associated with stroke (HR 0.66; 95% CI 0.42, 0.88) incidence, aortic pulse wave velocity, and C-reactive protein. The HDI was not associated with any single outcome.Higher DASH and AHEI-2010 scores were associated with lower CVD and stroke risk, and favourable cardiovascular health outcomes, suggesting that encouraging middle-aged men to comply with the dietary recommendations for a healthy diet may have important implications for future vascular disease and population health.
Keywords: Dietary adherence; Dietary approaches to stop hypertension score; Alternative healthy eating index-2010; Cardiovascular disease; Aortic pulse wave velocity

Inulin-type fructans are recognized as prebiotic dietary fibres and classified as non-digestible carbohydrates that do not contribute to glycaemia. The aim of the present studies was to investigate the glycaemic response (GR) and insulinaemic response (IR) to foods in which sucrose was partially replaced by inulin or oligofructose from chicory.In a double-blind, randomized, controlled cross-over design, 40–42 healthy adults consumed a yogurt drink containing oligofructose or fruit jelly containing inulin and the respective full-sugar variants. Capillary blood glucose and insulin were measured in fasted participants and at 15, 30, 45, 60, 90, and 120 min after starting to drink/eat. For each test food, the incremental area under the curve (iAUC) for glucose and insulin was calculated and the GR and IR determined.Consumption of a yogurt drink with oligofructose which was 20% reduced in sugars significantly lowered the glycaemic response compared to the full-sugar reference (iAUC120min 31.9 and 37.3 mmol/L/min, respectively; p < 0.05). A fruit jelly made with inulin and containing 30% less sugars than the full-sugar variant likewise resulted in a significantly reduced blood glucose response (iAUC120min 53.7 and 63.7 mmol/L/min, respectively; p < 0.05). In both studies, the postprandial insulin response was lowered in parallel (p < 0.05). The reduction of postprandial glycaemia was positively correlated to the proportion of sugars replaced by inulin-type fructans (p < 0.001).In conclusion, the studies confirmed that substitution of glycaemic sugars by inulin or oligofructose from chicory may be an effective strategy to reduce the postprandial blood glucose response to foods.
Keywords: Glycaemia; Insulin; Dietary fibre; Prebiotics; Sugar replacement

Research interactions between academia and food companies: how to improve transparency and credibility of an inevitable liaison by Andrea Poli; Franca Marangoni; Carlo V. Agostoni; Francesco Brancati; Lucio Capurso; Maria Laura Colombo; Andrea Ghiselli; Carlo La Vecchia; Enrico Molinari; Lorenzo Morelli; Marisa Porrini; Francesco Visioli; Gabriele Riccardi (1269-1273).