European Journal of Nutrition (v.54, #2)

Advances in water intake assessment by Jodi Dunmeyer Stookey; Juergen Koenig (9-10).

Good hydration is vital for good health and well-being. Until recently, there was little interest in collecting data on water and drink and beverage intake. However, there is increasing evidence that a low water intake or mild dehydration may be linked with the risk of chronic diseases. Accurate estimates of intake in populations are essential to explore these relationships. This will enable the identification of specific populations at the risk of low water intake and allow exposure assessment of potential contaminates and specific nutrients present in drinks and beverages. In addition, data from these population studies are used as the basis of national and international recommendations on water intake and to set and evaluate national health policies. For example, EFSA based their recommendations on data from population studies from 13 European countries. The range of intakes varied from 720 to 2621 mL/day; this diversity cannot be explained by environmental differences alone. However, this variability may, at least partially, be explained by the inconsistency in methodologies used as none of surveys used a dietary assessment tool validated for total water intake or beverage and drink intake. It is reasonable to suggest that this may result in incomplete data collection and it raises questions on the validity of the recommendations. The relationship between water consumption and health warrants further investigation, and robust methodologies are essential to ensure that these data are accurate and useful for setting public health priorities and policies.
Keywords: Water intake; Dietary assessment methodologies; Hydration; Fluid intake; Recommendations

Total fluid intake assessed with a 7-day fluid record versus a 24-h dietary recall: a crossover study in Indonesian adolescents and adults by Saptawati Bardosono; Romain Monrozier; Inge Permadhi; Nurul Ratna Mutu Manikam; Rizki Pohan; Isabelle Guelinckx (17-25).
To compare total fluid intake (TFI), defined as the sum of water and all other fluid types, assessed with a 24-h dietary (food and fluid) recall with mean TFI assessed with a 7-day fluid-specific record among adolescents and adults. This repeated cross-sectional study compared TFI as assessed by two fluid assessment instruments using a crossover approach. 290 adolescents (17.3 ± 0.8 years, 50 % boys) and 289 adults (43 ± 9.3 years, 50 % men) from Indonesia completed the study.Significant correlations were observed between fluid intake assessed with the 24-h recall and the 7-day fluid record (r = 0.333; p < 0.001). The Bland–Altman method, however, showed an underestimation (bias) of mean TFI by a 24-h recall when compared with the 7-day fluid record [mean difference (95 % CI) −382 mL (−299, −465); p < 0.001]. The mean difference also increased with increasing TFI: Mean difference for the lowest and highest quartiles of TFI was 139 versus −1265 mL/day. The 7-day fluid record recorded two (95 % CI −1.9, −2.4; p < 0.0001) extra drinking acts compared with the 24-h recall, whereas the mean volume per drinking act was significantly higher with the 24-h recall [mean difference (95 % CI) 39 mL (31, 47); p < 0.001].Compared with a 7-day fluid record, a 24-h dietary recall significantly underestimated TFI. Subjects recalled two less drinking acts, while estimating the volume consumed per drinking act to be larger. Since the adequate intakes for total water intake are based on median intakes observed in national surveys that most frequently used the 24-h recall method, they may potentially be underestimated.
Keywords: Water intake; Total fluid intake; 7-day fluid record; 24-h dietary recall; Indonesia

A cross-over study comparing an online versus a paper 7-day food record: focus on total water intake data and participant’s perception of the records by B. Monnerie; L. G. Tavoularis; I. Guelinckx; P. Hebel; T. Boisvieux; A. Cousin; L. Le Bellego (27-34).
To compare (1) fluid, food and nutrient intake obtained with a paper versus an online version of a 7-day food record and (2) user’s acceptability of both versions of the food record.A cross-over study was carried out in 2010 in France. A total of 246 participants aged 18–60 years reported their food and fluid intake using both versions of the 7-day food record, separated by a 7- to 14-day washout period. To help participants in estimating consumed portions, both versions of the food record were supported by a photographic booklet of standard portions and containers. At the end of the study protocol, participants completed a questionnaire designed to assess the acceptability of the two questionnaires.The reported water intake of fluids was significantly higher with the online version compared with the paper version (respectively 1348 ± 36 and 1219 ± 34 mL/day, p < 0.0001). No difference was found between methods in terms of energy intake and the consumption of most food categories, macro- and micronutrients. Furthermore, 77 % of the participants preferred the online method to the paper version.Fluid intake, but not food intake, reported with the online 7-day food record was higher in comparison with the paper version. In addition, the online version was preferred by users. In population surveys, the online record is therefore a relevant alternative, and even a preferred alternative in the case of fluid intake, to the paper record.
Keywords: Total water intake; Beverages; Fluids; Dietary record; Hydration

Total fluid intake and its determinants: cross-sectional surveys among adults in 13 countries worldwide by C. Ferreira-Pêgo; I. Guelinckx; L. A. Moreno; S. A. Kavouras; J. Gandy; H. Martinez; S. Bardosono; M. Abdollahi; E. Nasseri; A. Jarosz; N. Babio; J. Salas-Salvadó (35-43).
To evaluate the total fluid intake from drinking water and beverages in adult populations from different countries and assess the percentage of individuals complying with the European Food Safety Agency (EFSA) adequate intake (AI) of water from fluids.A total of 16,276 adults (7580 men and 8696 women) aged between 18 and 70 years (mean age 39.8 years) were randomly recruited from 13 different countries from three continents. Information about the total daily fluid intake (sum of drinking water and beverages) was collected using a 24-h fluid-specific record over seven consecutive days.Important differences in total fluid intake between countries were found; however, few differences between men and women were reported in most of the countries. Less than 50 % of the women and approximately 60 % of the men do not comply with the EFSA AI of water from fluids. Women were more than twice as likely as men to meet these AI (OR 2.15; 95 % CI 2.02–2.29). The odds of meeting the AI of water from fluids were lower in individuals over 50 years (OR 0.88; 95 % CI 0.80–0.96). Nine percent of the total population consumed less than half of the AI, 40.5 % between 50 and 100 %, and 50.5 % more than the AI.There were considerable differences in total fluid intake between countries but not between genders. Only 40 % of men and 60 % of women comply with the EFSA AI of water from fluids. Men and elderly individuals had an increased risk of not complying with this reference value.
Keywords: Fluid intake; Beverages; Adult population; EFSA adequate intake; Water; Liq.In7

Intake of water and different beverages in adults across 13 countries by I. Guelinckx; C. Ferreira-Pêgo; L. A. Moreno; S. A. Kavouras; J. Gandy; H. Martinez; S. Bardosono; M. Abdollahi; E. Nasseri; A. Jarosz; G. Ma; E. Carmuega; N. Babio; J. Salas-Salvadó (45-55).
To describe the intake of water and all other fluids and to evaluate the proportion of adults exceeding the World Health Organisation (WHO) recommendations on energy intake from free sugar, solely from fluids.A total of 16,276 adults (46 % men, mean age 39.8 years) were recruited in 13 countries from 3 continents. A 24-h fluid-specific record over 7 days was used for fluid assessment.In Spain, France, Turkey, Iran, Indonesia and China, fluid intake was characterised by a high contribution of water (47–78 %) to total fluid intake (TFI), with a mean water intake between 0.76 and 1.78 L/day, and a mean energy intake from fluids from 182 to 428 kcal/day. Between 11 and 49 % of adults exceeded the free sugar WHO recommendations, considering solely fluids. In Germany, UK, Poland and Japan, the largest contributors to TFI were hot beverages (28–50 %) and water (18–32 %). Mean energy intake from fluids ranged from 415 to 817 kcal/day, and 48–62 % of adults exceeded free sugar WHO recommendations. In Mexico, Brazil and Argentina, the contribution of juices and regular sugar beverages (28–41 %) was as important as the water contribution to TFI (17–39 %). Mean energy intake from fluids ranged 565–694 kcal/day, and 60–66 % of the adults exceeded the free sugar WHO recommendation.The highest volumes recorded in most of the countries were for water, mean energy intake from fluids was up to 694 kcal/day, and 66 % of adults exceeded the free sugar WHO recommendation solely by fluids. Actions to create an environment in favour of water consumption and reduce sugar intake from fluids therefore are warranted.
Keywords: Water; Beverages; Fluids; Adult population; WHO recommendation; Energy intake; Free sugars

Total fluid intake of children and adolescents: cross-sectional surveys in 13 countries worldwide by Iris Iglesia; Isabelle Guelinckx; Pilar M. De Miguel-Etayo; Esther M. González-Gil; Jordi Salas-Salvadó; Stavros A. Kavouras; Joan Gandy; Homero Martínez; Saptawati Bardosono; Morteza Abdollahi; Esmat Nasseri; Agnieszka Jarosz; Guansheng Ma; Esteban Carmuega; Isabelle Thiébaut; Luis A. Moreno (57-67).
To describe total fluid intake (TFI) according to socio-demographic characteristics in children and adolescents worldwide.Data of 3611 children (4–9 years) and 8109 adolescents (10–18 years) were retrieved from 13 cross-sectional surveys (47 % males). In three countries, school classes were randomly recruited with stratified cluster sampling design. In the other countries, participants were randomly recruited based on a quota method. TFI (drinking water and beverages of all kinds) was obtained with a fluid-specific record over 7 consecutive days. Adequacy was assessed by comparing TFI to 80 % of adequate intake (AI) for total water intake set by European Food Safety Authority. Data on height, weight and socio-economic level were collected in most countries.The mean (SD) TFI ranged from [1.32 (0.68)] to [1.35 (0.71)] L/day. Non-adherence to AIs for fluids ranged from 10 % (Uruguay) to >90 % (Belgium). Females were more likely to meet the AIs for fluids than males (4–9 years: 28 %, OR 0.72, p = 0.002; 10–18 years: 20 %, OR 0.80, p = 0.001), while adolescents were less likely to meet the AI than children (OR 1.645, p < 0.001 in males and OR 1.625, p < 0.001 in females).A high proportion of children and adolescents are at risk of an inadequate fluid intake. This risk is especially high in males and adolescents when compared with females or children categories. This highlights water intake among young populations as an issue of global concern.
Keywords: Fluid intake; Children; Adolescents; Worldwide; Dietary assessment; Hydration

Intake of water and beverages of children and adolescents in 13 countries by I. Guelinckx; I. Iglesia; J. H. Bottin; P. De Miguel-Etayo; E. M. González-Gil; J. Salas-Salvadó; S. A. Kavouras; J. Gandy; H. Martinez; S. Bardosono; M. Abdollahi; E. Nasseri; A. Jarosz; G. Ma; E. Carmuega; I. Thiébaut; Luis A. Moreno (69-79).
To describe the intake of water and all other beverages in children and adolescents in 13 countries of three continents. Data of 3611 children (4–9 years) and 8109 adolescents (10–17 years) were retrieved from 13 cross-sectional surveys (47 % males). In three countries, stratified cluster sampling design was applied to randomly recruit schools classes. A quota method was applied in the other countries to randomly recruit participants. Details on the intake of all fluid types were obtained with a fluid-specific record over 7 consecutive days. In the total sample, the highest mean intakes were observed for water (738 ± 567 mL/day), followed by milk (212 ± 209 mL/day), regular soft beverages (RSB) (168 ± 290 mL/day) and juices (128 ± 228 mL/day). Patterns characterized by a high contribution of water, RSB or hot beverages to total fluid intake were identified among the countries with close geographical location. Adolescents had a significantly lower milk intake and higher intake of RSB and hot beverages than children in most countries. The most consistent gender difference observed was that in both age groups males reported a significantly higher RSB consumption than females.On average, water was the fluid consumed in the largest volume by children and adolescents, but the intake of the different fluid types varied substantially between countries. Since the RSB intake was as large, or even larger, than water intake in some countries, undertaking actions to improve fluid intake habits of children and adolescents are warranted.
Keywords: Water; Beverages; Fluid intake; Children; Adolescents

Sister chromatid exchange (SCE) is a widely used sensitive cytogenetic biomarker of exposure to genotoxic and cancerogenic agents. Results of human monitoring studies and cytogenetic damage have revealed that biological effects of genotoxic exposures are influenced by confounding factors related to life-style. Vegetable and fruit consumption may play a role, but available results are not consistent. The purpose of the study was to investigate the effect of consumption of raw and cooked vegetables and fruits on SCE frequency.A total of 62 participants included colorectal cancer (CRC) patients, hospital-based controls and healthy laboratory workers. SCE frequency was assessed in blood lymphocytes. Frequency of vegetable and fruit consumption was gathered by structured semi-quantitative food frequency questionnaire.SCE frequency was lowest among hospital-based controls (4.4 ± 1.1), a bit higher in CRC patients (4.5 ± 1.0) and highest among laboratory workers (7.4 ± 1.2) (p < 0.05). Multivariable linear regression showed a significant inverse effect (b = −0.20) of raw vegetable consumption, but not so for intake of cooked vegetables and fruits.The results of the study have shown the beneficial effect of consumption of raw vegetables on disrupted replication of DNA measured by SCE frequency, implying protection against genotoxic agents. Further effort is required to verify the role of cooked vegetables and fruits.
Keywords: Biomarker; Sister chromatid exchange; Colorectal cancer; Controls; Raw and cooked vegetables; Fruits

Changes in the serum metabolite profile in obese children with weight loss by Thomas Reinehr; Barbara Wolters; Caroline Knop; Nina Lass; Christian Hellmuth; Ulrike Harder; Wolfgang Peissner; Simone Wahl; Harald Grallert; Jerzy Adamski; Thomas Illig; Cornelia Prehn; Zhonghao Yu; Rui Wang-Sattler; Berthold Koletzko (173-181).
Childhood obesity is an increasing problem and is accompanied by metabolic disturbances. Recently, we have identified 14 serum metabolites by a metabolomics approach (FIA-MS/MS), which showed altered concentrations in obese children as compared to normal-weight children. Obese children demonstrated higher concentrations of two acylcarnitines and lower levels of three amino acids, six acyl–alkyl phosphatidylcholines, and three lysophosphatidylcholines. The aim of this study was to analyze whether these alterations normalize in weight loss.We analyzed the changes of these 14 metabolites by the same metabolic kit as in our previous study in serum samples of 80 obese children with substantial weight loss (BMI-SDS reduction >0.5) and in 80 obese children with stable weight status all participating in a 1-year lifestyle intervention.In the children without weight change, no significant changes of metabolite concentrations could be observed. In children with substantial weight loss, glutamine, methionine, the lysophosphatidylcholines LPCaC18:1, LPCaC18:2, and LPCa20:4, as well as the acyl–alkyl phosphatidylcholine PCaeC36:2 increased significantly, while the acylcarnitines C12:1 and C16:1, proline, PCaeC34:1, PCaeC34:2, PCaeC34:3, PCaeC36:3, and PCaeC38:2 did not change significantly.The changes of glutamine, methionine, LPCaC18:1, LPCaC18:2, LPCa20:4, and PCaeC36:2 seem to be related to the changes of dieting or exercise habits in lifestyle intervention or to be a consequence of overweight since they normalized in weight loss. Further studies should substantiate our findings.
Keywords: Obesity; Childhood; Metabolomics; Metabolite profile; Weight loss; Lifestyle intervention

The effect of high-fat–high-fructose diet on skeletal muscle mitochondrial energetics in adult rats by Raffaella Crescenzo; Francesca Bianco; Paola Coppola; Arianna Mazzoli; Luisa Cigliano; Giovanna Liverini; Susanna Iossa (183-192).
To study the effect of isoenergetic administration to adult rats of high-fat or high-fat–high-fructose diet for 2 weeks on skeletal muscle mitochondrial energetic.Body and skeletal muscle composition, energy balance, plasma lipid profile and glucose tolerance were measured, together with mitochondrial functionality, oxidative stress and antioxidant defense.Rats fed high-fat–high-fructose diet exhibited significantly higher plasma triglycerides and non-esterified fatty acids, together with significantly higher plasma glucose and insulin response to glucose load. Skeletal muscle triglycerides and ceramide were significantly higher in rats fed high-fat–high-fructose diet. Skeletal muscle mitochondrial energetic efficiency and uncoupling protein 3 content were significantly higher, while adenine nucleotide translocase content was significantly lower, in rats fed high-fat or high-fat–high-fructose diet.The results suggest that a high-fat–high-fructose diet even without hyperphagia is able to increase lipid flow to skeletal muscle and mitochondrial energetic efficiency, with two detrimental effects: (a) energy sparing that contributes to the early onset of obesity and (b) reduced oxidation of fatty acids and lipid accumulation in skeletal muscle, which could generate insulin resistance.
Keywords: Insulin resistance; Mitochondrial coupling; Obesity; Ceramide

Hypolipidemic effect of dietary water-soluble protein extract from chicken: impact on genes regulating hepatic lipid and bile acid metabolism by Rita Vik; Bodil Bjørndal; Pavol Bohov; Trond Brattelid; Asbjørn Svardal; Ottar K. Nygård; Jan E. Nordrehaug; Jon Skorve; Rolf K. Berge (193-204).
Amount and type of dietary protein have been shown to influence blood lipids. The present study aimed to evaluate the effects of a water-soluble fraction of chicken protein (CP) on plasma and hepatic lipid metabolism in normolipidemic rats.Male Wistar rats were fed either a control diet with 20 % w/w casein as the protein source, or an experimental diet where casein was replaced with CP at 6, 14, or 20 % w/w for 4 weeks.Rats fed CP had markedly reduced levels of triacylglycerols (TAG) and cholesterol in both plasma and liver, accompanied by stimulated hepatic mitochondrial fatty acid oxidation and carnitine palmitoyltransferase 2 activity in the 20 % CP group compared to the control group. In addition, reduced activities and gene expression of hepatic enzymes involved in lipogenesis were observed. The gene expression of sterol regulatory element-binding transcription factor 1 was reduced in the 20 % CP-fed rats, whereas gene expression of peroxisome proliferator-activated receptor alpha was increased. Moreover, 6, 14, and 20 % CP-fed rats had significantly increased free carnitine and acylcarnitine plasma levels compared to control rats. The plasma methionine/glycine and lysine/arginine ratios were reduced in 20 % CP-treated rats. The mRNA level of ATP-binding cassette 4 was increased in the 20 % CP group, accompanied by the increased level of plasma bile acids.The present data suggest that the hypotriglyceridemic property of a water-soluble fraction of CP is primarily due to effects on TAG synthesis and mitochondrial fatty acid oxidation. The cholesterol-lowering effect by CP may be linked to increased bile acid formation.
Keywords: Chicken protein; Lipogenesis; β-Oxidation

Habitual consumption of coffee and green tea in relation to serum adipokines: a cross-sectional study by Ngoc Minh Pham; Akiko Nanri; Kazuki Yasuda; Kayo Kurotani; Keisuke Kuwahara; Shamima Akter; Masao Sato; Hitomi Hayabuchi; Tetsuya Mizoue (205-214).
Coffee and green tea consumption may be associated with circulating adipokines, but data are inconsistent, scarce or lacking. We examined the association of coffee and green tea consumption with serum adiponectin, leptin, visfatin, resistin and plasminogen activator inhibitor-1 (PAI-1) among a Japanese working population.The authors analyzed data (n = 509) from a cross-sectional survey among Japanese workers aged 20–68 years. Serum adipokines were measured using a Luminex suspension bead-based multiplexed array. Coffee and green tea consumption was assessed using a validated diet history questionnaire, and caffeine consumption from these beverages was estimated. Multiple regression analysis was performed with adjustment for potential confounding variables. Coffee consumption was significantly, inversely associated with leptin and PAI-1 (P for trend = 0.007 and 0.02, respectively); compared with subjects consuming <1 cup per day, those consuming ≥4 cups per day had 13 and 10 % lower means of leptin and PAI-1, respectively. Similar associations were observed for caffeine consumption (P for trend = 0.02 for both leptin and PAI-1). Additionally, we noted a significant positive association between coffee consumption and adiponectin in men (P for trend = 0.046), but not in women (P for trend = 0.43, P for interaction = 0.11). Moreover, there was a positive association between coffee consumption and resistin in current male smokers (P for trend = 0.01), but not in male non-smokers (P for trend = 0.35, P for interaction = 0.11). Green tea consumption was not associated with any adipokine. Higher consumption of coffee and caffeine but not green tea was associated with lower serum levels of leptin and PAI-1 in Japanese adults.
Keywords: Coffee; Green tea; Adipokines; Japanese

Relation of dietary glycemic load with ischemic and hemorrhagic stroke: a cohort study in Greece and a meta-analysis by Marta Rossi; Federica Turati; Pagona Lagiou; Dimitrios Trichopoulos; Carlo La Vecchia; Antonia Trichopoulou (215-222).
High glycemic load (GL) has been associated with excess stroke risk. Data suggest a different role of diet in the etiology of ischemic and hemorrhagic stroke.We analyzed data from 19,824 participants of the Greek cohort of the population-based European Prospective Investigation into Cancer and nutrition (EPIC), who were free of cardiovascular diseases, cancer, and diabetes at baseline and had not developed diabetes. Diet was assessed at enrollment through a validated, interviewer-administered semi-quantitative food frequency questionnaire. The average daily GL was derived using standard tables. We also conducted a meta-analysis on GL and stroke (overall, ischemic and hemorrhagic), using random-effects models.In the Greek EPIC cohort, 304 incident stroke cases were identified (67 ischemic, 49 hemorrhagic). Using Cox proportional hazards regression models adjusted for potential confounders, the hazard ratios for the highest versus the lowest GL tertiles were 1.07 [95 % confidence interval (CI) 0.74–1.54] for overall stroke, 1.55 (95 % CI 0.72–3.36) for ischemic and 0.48 (95 % CI 0.18–1.25) for hemorrhagic stroke (p-heterogeneity <0.01). The meta-analysis, including a total of 3,088 incident cases and 247 deaths from stroke (1,469 cases and 126 deaths ischemic; 576 cases and 94 deaths hemorrhagic), estimated pooled relative risks for the highest versus the lowest GL levels of 1.23 (95 % CI 1.07–1.41) for overall, 1.35 (95 % CI 1.06–1.72) for ischemic, and 1.09 (95 % CI 0.81–1.47) for hemorrhagic stroke (p-heterogeneity = 0.275).This study indicates that GL is an important determinant of the more common ischemic—though not of the hemorrhagic—stroke.
Keywords: Diet; Glycemic load; Stroke; Cohort study; Meta-analysis

GSH protects against oxidative stress and toxicity in VL-17A cells exposed to high glucose by S. Mathan Kumar; Kavitha Swaminathan; Dahn L. Clemens; Aparajita Dey (223-234).
The deficiency of glutathione (GSH) has been linked to several diseases. The study investigated the role of GSH as a protective factor against hyperglycemia-mediated injury in VL-17A cells treated with 50 mM glucose.The cell viability and different oxidative stress parameters including glyoxalase I activity were measured.GSH supplementation with 2 mM N-acetyl cysteine (NAC) or 0.1 mM ursodeoxycholic acid (UDCA) increased the viability, GSH level and the GSH-dependent glyoxalase I activity in 50 mM glucose-treated VL-17A cells. Further, pretreatment of 50 mM glucose-treated VL-17A cells with NAC or UDCA decreased oxidative stress (levels of reactive oxygen species and protein carbonylation), apoptosis (caspase 3 activity and annexin V–propidium iodide positive cells) and glutathionylated protein formation, a measure of oxidative stress. GSH depletion with 0.4 mM buthionine sulfoximine (BSO) or 1 mM diethyl maleate (DEM) potentiated the decrease in viability, glyoxalase I activity and increase in oxidative stress and apoptosis, with decreased GSH levels in 50 mM glucose-treated VL-17A cells.Thus, changes in GSH levels with exogenous agents such as NAC, UDCA, BSO or DEM modulate hyperglycemia-mediated injury in a cell model of VL-17A liver cells.
Keywords: Glutathione; Hyperglycemia; Liver; Reactive oxygen species; Injury

High plasma folate is negatively associated with leukocyte telomere length in Framingham Offspring cohort by Ligi Paul; Paul F. Jacques; Abraham Aviv; Ramachandran S. Vasan; Ralph B. D’Agostino; Daniel Levy; Jacob Selhub (235-241).
Shortening of telomeres, the protective structures at the ends of eukaryotic chromosomes, is associated with age-related pathologies. Telomere length is influenced by DNA integrity and DNA and histone methylation. Folate plays a role in providing precursors for nucleotides and methyl groups for methylation reactions and has the potential to influence telomere length.We determined the association between leukocyte telomere length and long-term plasma folate status (mean of 4 years) in Framingham Offspring Study (n = 1,044, females = 52.1 %, mean age 59 years) using data from samples collected before and after folic acid fortification. Leukocyte telomere length was determined by Southern analysis and fasting plasma folate concentration using microbiological assay.There was no significant positive association between long-term plasma folate and leukocyte telomere length among the Framingham Offspring Study participants perhaps due to their adequate folate status. While the leukocyte telomere length in the second quintile of plasma folate was longer than that in the first quintile, the difference was not statistically significant. The leukocyte telomere length of the individuals in the fifth quintile of plasma folate was shorter than that of those in the second quintile by 180 bp (P < 0.01). There was a linear decrease in leukocyte telomere length with higher plasma folate concentrations in the upper four quintiles of plasma folate (P for trend = 0.001). Multivitamin use was associated with shorter telomeres in this cohort (P = 0.015).High plasma folate status possibly resulting from high folic acid intake may interfere with the role of folate in maintaining telomere integrity.
Keywords: Telomere length; Folate; Multivitamins; Folic acid fortification

Poultry intake has been inconsistently associated with incidence or mortality of colorectal cancer (CRC) in epidemiologic studies. The purpose of this study was to assess their relationships by performing a dose–response meta-analysis.We conducted a search of PubMed database between January 1966 and July 2013 for prospective studies that reported relative risks (RRs) with 95 % confidence interval (CIs) of CRC for at least three categories of poultry intake. Dose–response relationships were examined with the generalized least-squares trend estimation. Study-specific results were pooled with a random-effects model. Subgroup, sensitivity, and meta-regression analyses were also conducted to explore heterogeneity.Sixteen studies on poultry intake and CRC incidence, and four studies regarding poultry intake and CRC mortality were identified. These studies involved a total of 13,949 incident CRC cases and 983 CRC deaths. The RRs of CRC for higher compared with lower intake of poultry were reported in these studies, and the reported levels of poultry intake varied substantially. Results of the dose–response meta-analysis conferred a RR of 0.89 (95 % CI 0.81–0.97) for an increase in poultry intake of 50 g/day. The results were not sensitive to any individual studies and were similar for colon and rectal cancer. Poultry intake was not associated with CRC mortality (RR for 50 g/day = 0.97, 95 % CI 0.79–1.20).This meta-analysis indicates that poultry intake may be moderately associated with reduced incidence of CRC.
Keywords: Meat; Poultry; Cohort studies; Colorectal cancer; Meta-analysis

Randomized controlled trial of oral glutathione supplementation on body stores of glutathione by John P. Richie Jr.; Sailendra Nichenametla; Wanda Neidig; Ana Calcagnotto; Jeremy S. Haley; Todd D. Schell; Joshua E. Muscat (251-263).
Glutathione (GSH), the most abundant endogenous antioxidant, is a critical regulator of oxidative stress and immune function. While oral GSH has been shown to be bioavailable in laboratory animal models, its efficacy in humans has not been established. Our objective was to determine the long-term effectiveness of oral GSH supplementation on body stores of GSH in healthy adults. A 6-month randomized, double-blinded, placebo-controlled trial of oral GSH (250 or 1,000 mg/day) on GSH levels in blood, erythrocytes, plasma, lymphocytes and exfoliated buccal mucosal cells was conducted in 54 non-smoking adults. Secondary outcomes on a subset of subjects included a battery of immune markers.GSH levels in blood increased after 1, 3 and 6 months versus baseline at both doses. At 6 months, mean GSH levels increased 30–35 % in erythrocytes, plasma and lymphocytes and 260 % in buccal cells in the high-dose group (P < 0.05). GSH levels increased 17 and 29 % in blood and erythrocytes, respectively, in the low-dose group (P < 0.05). In most cases, the increases were dose and time dependent, and levels returned to baseline after a 1-month washout period. A reduction in oxidative stress in both GSH dose groups was indicated by decreases in the oxidized to reduced glutathione ratio in whole blood after 6 months. Natural killer cytotoxicity increased >twofold in the high-dose group versus placebo (P < 0.05) at 3 months.These findings show, for the first time, that daily consumption of GSH supplements was effective at increasing body compartment stores of GSH.
Keywords: Glutathione; Supplementation; Antioxidant; Immune function

Autophagy inhibitor 3-methyladenine potentiates apoptosis induced by dietary tocotrienols in breast cancer cells by Anh Thu Tran; Malathi Ramalinga; Habib Kedir; Robert Clarke; Deepak Kumar (265-272).
Tocomin® represents commercially available mixture of naturally occurring tocotrienols (T3s) and tocopherols extracted from palm oil/palm fruits that possess powerful antioxidant, anticancer, neuro/cardioprotective and cholesterol-lowering properties. Cellular autophagy represents a defense mechanism against oxidative stress and several anticancer compounds. Recently, we reported that T3s induce apoptosis and endoplasmic reticulum stress in breast cancer cells.We studied the effects of Tocomin® on MCF-7 and MDA-MB 231 breast cancer cells and non-tumor MCF-10A cells.Tocomin® inhibited cell proliferation and induced apoptosis in both MCF-7 and MDA-MB 231 breast cancer cell lines without affecting the viability of MCF-10A cells. We also showed that Tocomin® negatively modulates phosphoinositide 3-kinase and mTOR pathways and induces cytoprotective autophagic response in triple negative MDA-MB 231 cells. Lastly, we demonstrate that autophagy inhibitor 3-methyladenine (3-MA) potentiated the apoptosis induced by Tocomin® in MDA-MB 231 cells.Together, our data indicate anticancer effects of Tocomin® in breast cancer cells, which is potentiated by the autophagy inhibitor 3-MA.
Keywords: Vitamin E; Tocomin® ; Tocotrienols; Breast cancer; Apoptosis; Autophagy

Associations between a posteriori defined dietary patterns and bone mineral density in adolescents by Teresa Monjardino; Raquel Lucas; Elisabete Ramos; Carla Lopes; Rita Gaio; Henrique Barros (273-282).
Dietary pattern analysis may uncover the joint effects of multiple dietary components on bone health, but such research is scarce and targets mostly adults.We quantified prospective associations between dietary patterns and bone mineral density (BMD) in 1,007 adolescents of a cohort born in 1990 and recruited at schools in Porto during the 2003/2004 school year. Forearm BMD was measured using dual-energy X-ray absorptiometry. Participants’ dietary patterns were classified “Healthier”, “Dairy products”, “Fast food and sweets” and “Lower intake” according to previously identified patterns obtained in a larger sample of 1,489 participants using the K-means method. Using dietary patterns at 13 years old as the main exposure, associations were estimated cross-sectionally (with BMD at the age of 13) and prospectively (with annual BMD variation between 13 and 17 years), using linear regression coefficients adjusted for height, weight, energy intake and, in girls, for menarche age.No significant associations between the a posteriori dietary patterns identified and mean BMD at 13 were found. However, among girls, adherence to a pattern characterized by low intake of energy and all food groups was negatively associated with annual BMD variation between 13 and 17 years [adjusted coefficient (95 % CI) −0.451 (−0.827; −0.074) mg·cm−2·year−1].Although results showed that, in girls, adherence to a “Lower intake” dietary pattern is associated with lower annual BMD variation throughout adolescence, overall, there were no consistent associations between dietary patterns and forearm BMD in adolescents.
Keywords: Dietary patterns; Bone density; Cohort study; Adolescence

Serum ghrelin levels and gender-related indices of body composition in prepubertal children: a cross-sectional study by Minoo Bagheri; Sara Ansari; Gity Sotoudeh; Mahmood Mahmoudi; John R. Speakman; Kurosh Djafarian (283-290).
A wide variety of functions has been attributed to ghrelin, a peptide hormone secreted in the stomach. The objective of the study was to assess the association of ghrelin concentrations with body composition among Iranian children.In this study, blood samples of 57 boys and 54 girls aged 6–10 were collected to measure ghrelin levels. Fat mass (FM) and fat-free mass (FFM) were examined by body composition analyzer. Actigraph GT3X was administered to assess children’s physical activity and sleep. Data were analyzed using linear regression models.All measured parameters did not differ between genders except for sleep time which was higher and sleep efficacy which was lower in boys compared with girls. None of the FM and FFM indices studied in boys was significantly associated with ghrelin levels. In girls, however, ghrelin concentrations were significantly associated with FM (β = 0.04, P = 0.01), fat mass index (β = 0.07, P = 0.008), and fat-free mass index (β = 0.08, P = 0.04) and near-significantly associated with FFM (β = 0.03, P = 0.09) after adjusting for age, physical activity, sleep, and dietary intake.Girls with higher ghrelin levels were more likely to have increased total FM and FFM. Conversely, body composition was not associated with ghrelin levels in boys. Consequently, ghrelin may influence the gender-related differences of body composition during childhood in girls. But, further study is needed to confirm our findings.
Keywords: Ghrelin; Fat mass; Fat-free mass; Body composition; Children

Effect of walnut oil on hyperglycemia-induced oxidative stress and pro-inflammatory cytokines production by Lucia Laubertová; Katarína Koňariková; Helena Gbelcová; Zdeňka Ďuračková; Ingrid Žitňanová (291-299).
In this study, we focused on the effect of hyperglycemia on the generation of reactive oxygen species and on the release of pro-inflammatory cytokines in the human monocytic cell line (U937). We also monitored potential anti-inflammatory effects of walnut oil as well as its protective effect against oxidative damage to biopolymers (DNA and proteins).We cultured U937 cells under normoglycemic or hyperglycemic conditions for 72 h, in the absence or presence of walnut oil. We detected cell proliferation by the MTT test. To determine the antioxidant status of cells, we used the trolox equivalent antioxidant capacity method. We determined the activity of superoxide dismutase (SOD) spectrophotometrically, the oxidative damage to DNA by an enzyme-modified comet assay, and the oxidative damage to proteins by the marker—protein carbonyls and the levels of pro-inflammatory cytokines by the ELISA method.Hyperglycemia reduced the antioxidant capacity of cells, induced oxidative damage to DNA, and increased the release of pro-inflammatory cytokines. It had no effect on cell proliferation, SOD activity, nor oxidative damage to proteins. Walnut oil significantly increased the antioxidant capacity of cells as well as SOD activity on the second and third day of incubation, but had no effect on cell proliferation and showed no protective effect against oxidative damage to DNA and proteins. The walnut oil showed both anti-inflammatory and pro-inflammatory properties depending on its concentration and time of its incubation with the monocytic cell line.Our in vitro results indicate that walnut oil can diminish oxidative stress with its antioxidant properties. However, we could not confirm its protective effect against oxidative damage to DNA and proteins.
Keywords: Hyperglycemia; Oxidative stress; Cytokines; Plant oil; Cell line

Astaxanthin from shrimp by-products ameliorates nephropathy in diabetic rats by Assaâd Sila; Zohra Ghlissi; Zeineb Kamoun; Mohamed Makni; Moncef Nasri; Ali Bougatef; Zouheir Sahnoun (301-307).
This study investigated the hypoglycemic and antioxidant effects of shrimp astaxanthin on the kidney of alloxan-induced diabetic rats.Animals were distributed into four groups of six rats each: a control group (C), a diabetic group (D), a diabetic group supplemented with Astaxanthin (D+As) dissolved in olive oil and a diabetic group supplemented with olive oil (D+OO). In vitro antidiabetic effect was tested in plasma and kidney tissue. The group D of rats showed significant (P < 0.05) increase of glycemia, creatinine, urea and uric acid levels compared to those of the control group (C). Moreover, plasma and kidney malondialdehyde (MDA) and protein carbonyl (PCO) levels for the rats of the group D were significantly increased compared to the control group. Contrariwise, antioxidant enzyme activities, such as catalase (EC 1.11.1.6), superoxide dismutase (EC 1.15.1.1) and non-enzymatic levels of reduced glutathione, were significantly (P < 0.05) decreased in the plasma and kidney of diabetic rats compared to the control ones. The astaxanthin supplementation in rats diet improved the antioxidant enzyme activities and significantly decreased the MDA and PCO levels compared to diabetic rats. Indeed, no significant (P ≥ 0.05) improvement was observed for the fourth group (D+OO) compared to the control group (C). Histological analysis of kidney showed glomerular hypertrophy and tubular dilatation for the diabetic rats. For D+As rats, these histopathological changes were less prominent.Our results suggest that shrimp astaxanthin may play an important role in reduction of oxidative damage and could prevent pathological changes in diabetic rats suggesting promising application of shrimp astaxanthin in diabet treatment.
Keywords: Astaxanthin; Diabetes; Oxidative stress; Lipid peroxidation; Nephropathy

Dietary patterns and cognitive function in Korean older adults by Jihye Kim; Areum Yu; Bo Youl Choi; Jung Hyun Nam; Mi Kyung Kim; Dong Hoon Oh; Kirang Kim; Yoon Jung Yang (309-318).
The objectives of this study were to identify major dietary patterns and to investigate the association between dietary patterns and cognitive function in older adults.This is a cross-sectional study. The data from the Korean Multi-Rural Communities Cohort Study, which is a part of the Korean Genome Epidemiology Study, were used. There were 806 (340 men and 466 women) subjects aged ≥60 years. Usual dietary intake was assessed using a quantitative food frequency questionnaire with 106 food items. Cognitive function was assessed using the Korean version Mini-Mental State Examination (MMSE-KC). We conducted factor analysis using the principal component analysis method to identify the major dietary patterns. The association between major dietary patterns and cognitive function was investigated by logistic regression analysis.Three major dietary patterns were identified and assigned descriptive names based on the food items with high loadings: “prudent” pattern, “bread, egg, and dairy” pattern, and “white rice only” pattern. As the white rice only pattern scores increased, a significant decreasing trend for MMSE-KC scores was observed after adjusting for covariates. The bread, egg, and dairy pattern was inversely related to the risk of cognitive impairment, and the white rice only pattern was positively associated with the risk of cognitive impairment.This study suggests that specific dietary patterns were significantly associated with cognitive impairment in older adults. In particular, like the white rice only pattern, a rice-centered diet without well-balanced meals may increase the risk of cognitive impairment. However, since our study is a cross-sectional design, the possibility of reverse causality should be considered.
Keywords: Dietary patterns; Cognitive function; Older adults; Cognitive impairment

Protective effect of KI in mtDNA in porcine thyroid: comparison with KIO3 and nDNA by Malgorzata Karbownik-Lewinska; Jan Stepniak; Magdalena Milczarek; Andrzej Lewinski (319-323).
Iodine, bivalent iron (Fe2+), and hydrogen peroxide (H2O2), all significantly affecting the red-ox balance, are required for thyroid hormone synthesis. Intracellular iodine excess (≥10−3 M) transiently blocks thyroid hormonogenesis (an adaptive mechanism called Wolff–Chaikoff effect). The aim of the study was to evaluate the effects of iodine, used as potassium iodide (KI) or potassium iodate (KIO3), in concentrations corresponding to those typical for Wolff–Chaikoff effect, on the level of oxidative damage to nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) isolated from porcine thyroid under basal conditions and in the presence of Fenton reaction (Fe2++H2O2 → Fe3++·OH + OH) substrates.Thyroid nDNA and mtDNA were incubated in the presence of either KI or KIO3 (2.5–50 mM), without/with FeSO4 (30 µM) + H2O2 (0.5 mM). Index of DNA damage, i.e., 8-oxo-7,8-dihydro-2′-deoxyguanosine, was measured by HPLC.Neither KI nor KIO3 increased the basal level of 8-oxodG in both nDNA and mtDNA. KI—in all used concentrations—completely prevented the damaging effect of Fenton reaction substrates in mtDNA, and it partially prevented this damage in nDNA. KIO3 partially prevented Fe2++H2O2-induced oxidative damage in both DNA only in its highest used concentrations (≥25 mM).Without additional prooxidative abuse, both iodine compounds, i.e., KI and KIO3, seem to be safe in terms of their potential oxidative damage to DNA in the thyroid. The superiority of KI over KIO3 relies on its stronger protective effects against oxidative damage to mtDNA, which constitutes an argument for its preferential utility in iodine prophylaxis.
Keywords: Potassium iodide; Potassium iodate; Nuclear DNA; Mitochondrial DNA; Thyroid