European Journal of Nutrition (v.55, #3)

Increasing vegetable intakes: rationale and systematic review of published interventions by Katherine M. Appleton; Ann Hemingway; Laure Saulais; Caterina Dinnella; Erminio Monteleone; Laurence Depezay; David Morizet; F. J. Armando Perez-Cueto; Ann Bevan; Heather Hartwell (869-896).
While the health benefits of a high fruit and vegetable consumption are well known and considerable work has attempted to improve intakes, increasing evidence also recognises a distinction between fruit and vegetables, both in their impacts on health and in consumption patterns. Increasing work suggests health benefits from a high consumption specifically of vegetables, yet intakes remain low, and barriers to increasing intakes are prevalent making intervention difficult. A systematic review was undertaken to identify from the published literature all studies reporting an intervention to increase intakes of vegetables as a distinct food group.Databases—PubMed, PsychInfo and Medline—were searched over all years of records until April 2015 using pre-specified terms.Our searches identified 77 studies, detailing 140 interventions, of which 133 (81 %) interventions were conducted in children. Interventions aimed to use or change hedonic factors, such as taste, liking and familiarity (n = 72), use or change environmental factors (n = 39), use or change cognitive factors (n = 19), or a combination of strategies (n = 10). Increased vegetable acceptance, selection and/or consumption were reported to some degree in 116 (83 %) interventions, but the majority of effects seem small and inconsistent.Greater percent success is currently found from environmental, educational and multi-component interventions, but publication bias is likely, and long-term effects and cost-effectiveness are rarely considered. A focus on long-term benefits and sustained behaviour change is required. Certain population groups are also noticeably absent from the current list of tried interventions.
Keywords: Vegetables; Interventions; Systematic review; Published literature

Functional gastrointestinal symptoms such as abdominal pain, bloating, distension, constipation, diarrhea and flatulence have been noted in patients with irritable bowel syndrome (IBS) or inflammatory bowel disease (IBD). The diversity of symptoms has meant that finding an effective treatment has been challenging with most treatments alleviating only the primary symptom. A novel treatment option for IBS and IBD currently generating much excitement is the low fermentable, oligo-, di-, mono-saccharides and polyol (FODMAP) diet. The aim of this meta-analysis was to determine the evidence of the efficacy of such a diet in the treatment of functional gastrointestinal symptoms.Electronic databases were searched through to March 2015 to identify relevant studies. Pooled odds ratios (ORs) and 95 % confidence intervals were calculated for the effect of a low FODMAP diet on the reduction in IBS [Symptoms Severity Score (SSS)] score and increase in IBS quality of life (QOL) score for both randomized clinical trials (RCTs) and non-randomized interventions using a random-effects model.Six RCTs and 16 non-randomized interventions were included in the analysis. There was a significant decrease in IBS SSS scores for those individuals on a low FODMAP diet in both the RCTs (OR 0.44, 95 % CI 0.25–0.76; I 2 = 35.52, p = 0.00) and non-randomized interventions (OR 0.03, 95 % CI 0.01–0.2; I 2 = 69.1, p = 0.02). In addition, there was a significant improvement in the IBS-QOL score for RCTs (OR 1.84, 95 % CI 1.12–3.03; I 2 = 0.00, p = 0.39) and for non-randomized interventions (OR 3.18, 95 % CI 1.60–6.31; I 2 = 0.00, p = 0.89). Further, following a low FODMAP diet was found to significantly reduce symptom severity for abdominal pain (OR 1.81, 95 % CI 1.13–2.88; I 2 = 0.00, p = 0.56), bloating (OR 1.75, 95 % CI 1.07–2.87; I 2 = 0.00, p = 0.45) and overall symptoms (OR 1.81, 95 % CI 1.11–2.95; I 2 = 0.00, p = 0.4) in the RCTs. In the non-randomized interventions similar findings were observed.The present meta-analysis supports the efficacy of a low FODMAP diet in the treatment of functional gastrointestinal symptoms. Further research should ensure studies include dietary adherence, and more studies looking at greater number of patients and long-term adherence to a low FODMAP diet need to be conducted.
Keywords: FODMAP; Diet; Functional gastrointestinal disorders; FGID; Symptoms; Meta-analysis

Validation of a urine color scale for assessment of urine osmolality in healthy children by Stavros A. Kavouras; Evan C. Johnson; Dimitris Bougatsas; Giannis Arnaoutis; Demosthenes B. Panagiotakos; Erica Perrier; Alexis Klein (907-915).
Urine color (UC) is a practical tool for hydration assessment. The technique has been validated in adults, but has not been tested in children.The purpose of the study was to test the validity of the urine color scale in young, healthy boys and girls, as a marker of urine concentration, investigate its diagnostic ability of detecting hypohydration and examine the ability of children to self-assess UC.A total of 210 children participated (age: 8–14 years, body mass: 43.4 ± 12.6 kg, height: 1.49 ± 0.13 m, body fat: 25.2 ± 7.8 %). Data collection included: two single urine samples (first morning and before lunch) and 24-h sampling. Hydration status was assessed via urine osmolality (UOsmo) and UC via the eight-point color scale.Mean UC was 3 ± 1 and UOsmo 686 ± 223 mmol kg−1. UC displayed a positive relationship as a predictor of UOsmo (R 2: 0.45, P < 0.001). Based on the receiver operating curve, UC has good overall classification ability for the three samples (area under the curve 85–92 %), with good sensitivity (92–98 %) and specificity (55–68 %) for detecting hypohydration. The overall accuracy of the self-assessment of UC in the morning or the noon samples ranged from 67 to 78 %. Further threshold analysis indicated that the optimal self-assessed UC threshold for hypohydration was ≥4.The classical eight-point urine color scale is a valid method to assess hydration in children of age 8–14 years, either by researchers or self-assessment.
Keywords: Hydration status; Children; Hydration assessment; Dehydration; Hypohydration markers

Indirect effects of a high-protein maternal diet are not well understood. In this study, we analyzed short-term and sustainable effects of a prenatal versus early postnatal maternal high-protein diet on growth and hepatic gene expression in mouse offspring.Dams were exposed to an isoenergetic high-protein (HP, 40 % w/w) diet during pregnancy or lactation. Growth and hepatic expression profiles of male offspring were evaluated directly after weaning and 150 days after birth. Offspring from two dietary groups, high-protein diet during pregnancy and control diet during lactation (HPC), and control diet during pregnancy and high-protein diet during lactation (CHP), were compared with offspring (CC) from control-fed dams.Maternal CHP treatment was associated with sustained offspring growth retardation, but decreased numbers of affected hepatic genes in adults compared to weanlings. In contrast, offspring of the HPC group did not show persistent effects on growth parameters, but the number of affected hepatic genes was even increased at adult age. In both dietary groups, however, only a small subset of genes was affected in weanlings as well as in adults.We conclude that (1) prenatal and early postnatal maternal HP diet caused persistent, but (2) different effects and partially complementary trends on growth characteristics and on the hepatic transcriptome and associated pathways and that (3) only a small number of genes and associated upstream regulators might be involved in passing early diet-induced imprints to adulthood.
Keywords: High-protein diet; Microarray analysis; Pregnancy; Lactation

Medium-chain triglyceride ameliorates insulin resistance and inflammation in high fat diet-induced obese mice by Shanshan Geng; Weiwei Zhu; Chunfeng Xie; Xiaoting Li; Jieshu Wu; Zhaofeng Liang; Wei Xie; Jianyun Zhu; Cong Huang; Mingming Zhu; Rui Wu; Caiyun Zhong (931-940).
The aim of the present study was to investigate the in vivo effects of dietary medium-chain triglyceride (MCT) on inflammation and insulin resistance as well as the underlying potential molecular mechanisms in high fat diet-induced obese mice.Male C57BL/6J mice (n = 24) were fed one of the following three diets for a period of 12 weeks: (1) a modified AIN-76 diet with 5 % corn oil (normal diet); (2) a high-fat control diet (17 % w/w lard and 3 % w/w corn oil, HFC); (3) an isocaloric high-fat diet supplemented with MCT (17 % w/w MCT and 3 % w/w corn oil, HF–MCT). Glucose metabolism was evaluated by fasting blood glucose levels and intraperitoneal glucose tolerance test. Insulin sensitivity was evaluated by fasting serum insulin levels and the index of homeostasis model assessment-insulin resistance. The levels of serum interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-α were measured by ELISA, and hepatic activation of nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathways was determined using western blot analysis.Compared to HFC diet, consumption of HF-MCT did not induce body weight gain and white adipose tissue accumulation in mice. HFC-induced increases in serum fasting glucose and insulin levels as well as glucose intolerance were prevented by HF-MCT diet. Meanwhile, HF-MCT resulted in significantly lower serum IL-6 level and higher IL-10 level, and lower expression levels of inducible nitric oxide synthase and cyclooxygenase-2 protein in liver tissues when compared to HFC. In addition, HF-MCT attenuated HFC-triggered hepatic activation of NF-κB and p38 MAPK.Our study demonstrated that MCT was efficacious in suppressing body fat accumulation, insulin resistance, inflammatory response, and NF-κB and p38 MAPK activation in high fat diet-fed mice. These data suggest that MCT may exert beneficial effects against high fat diet-induced insulin resistance and inflammation.
Keywords: Medium-chain triglyceride; Insulin resistance; Inflammation; NF-κB; p38 MAPK

Labrador tea (Rhododendron groenlandicum) attenuates insulin resistance in a diet-induced obesity mouse model by Meriem Ouchfoun; Hoda M. Eid; Lina Musallam; Antoine Brault; Shilin Li; Diane Vallerand; John T. Arnason; Pierre S. Haddad (941-954).
Using a diet-induced obesity (DIO) mouse model, we investigated the antidiabetic effect of Labrador tea [Rhododendron groenlandicum (Oeder) Kron and Judd], a beverage and medicinal tea used by the Cree Nations of northern Quebec.C57BL6 mice were divided into five groups and given standard chow (~4 % of lipids) or high-fat diet (~35 % of lipids) for 8 weeks until they became obese and insulin resistant. Treatment began by adding the plant extract at three doses (125, 250 and 500 mg/kg) to the high-fat diet for another 8 weeks. At the end of the study, insulin-sensitive tissues (liver, skeletal muscle, adipose tissue) were collected to investigate the plant’s molecular mechanisms.Labrador tea significantly reduced blood glucose (13 %), the response to an oral glucose tolerance test (18.2 %) and plasma insulin (65 %) while preventing hepatic steatosis (42 % reduction in hepatic triglyceride levels) in DIO mice. It stimulated insulin-dependent Akt pathway (55 %) and increased the expression of GLUT4 (53 %) in skeletal muscle. In the liver, Labrador tea stimulated the insulin-dependent Akt and the insulin-independent AMP-activated protein kinase pathways. The improvement in hepatic steatosis observed in DIO-treated mice was associated with a reduction in inflammation (through the IKK α/β) and a decrease in the hepatic content of SREBP-1 (39 %).Labrador tea exerts potential antidiabetic action by improving insulin sensitivity and mitigating high-fat diet-induced obesity and hyperglycemia. They validate the safety and efficacy of this plant, a promising candidate for culturally relevant complementary treatment in Cree diabetics.
Keywords: Labrador tea; AMPK; GLUT4; SREBP-1; Natural health products; Diabetes

Fruit peel polyphenols demonstrate substantial anti-tumour effects in the model of breast cancer by Peter Kubatka; Andrea Kapinová; Martin Kello; Peter Kruzliak; Karol Kajo; Desanka Výbohová; Silvia Mahmood; Radovan Murin; Tischlerová Viera; Ján Mojžiš; Anthony Zulli; Martin Péč; Marián Adamkov; Monika Kassayová; Bianka Bojková; Nadežda Stollárová; Dušan Dobrota (955-965).
Fruit and vegetable intake is inversely correlated with cancer; thus, it is proposed that an extract of phytochemicals as present in whole fruits, vegetables, or grains may have anti-carcinogenic properties. Thus, the anti-tumour effects of fruit peel polyphenols (Flavin7) in the chemoprevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis in female rats were evaluated.Lyophilized substance of Flavin7 (F7) was administered at two concentrations of 0.3 and 3 % through diet. The experiment was terminated 14 weeks after carcinogen administration, and mammary tumours were removed and prepared for histopathological and immunohistochemical analysis. In addition, using an in vitro cytotoxicity assay, apoptosis and proliferation after F7 treatment in human breast adenocarcinoma (MCF-7) cells were performed. High-dose F7 suppressed tumour frequency by 58 % (P < 0.001), tumour incidence by 24 % (P < 0.05), and lengthened latency by 8 days (P > 0.05) in comparison with the control rats, whereas lower dose of F7 was less effective. Histopathological analysis of tumours showed significant decrease in the ratio of high-/low-grade carcinomas after high-dose F7 treatment. Immunohistochemical analysis of rat carcinoma cells in vivo found a significant increase in caspase-3 expression and significant decrease in Bcl-2, Ki67, and VEGFR-2 expression in the high-dose group. Both doses demonstrated significant positive effects on plasma lipid metabolism in rats. F7 significantly decreased survival of MCF-7 cells in vitro in MTT assay by dose- and time-dependent manner compared to control. F7 prevented cell cycle progression by significant enrichment in G1 cell populations. Incubation with F7 showed significant increase in the percentage of annexin V-/PI-positive MCF-7 cells and DNA fragmentation. Our results reveal a substantial tumour-suppressive effect of F7 in the breast cancer model. We propose that the effects of phytochemicals present in this fruit extract are responsible for observed potent anti-cancer activities.
Keywords: Mammary carcinogenesis; Rat; Fruit polyphenols; Angiogenesis; Apoptosis; Cell proliferation; MCF-7

Longitudinal associations of serum fatty acid composition with type 2 diabetes risk and markers of insulin secretion and sensitivity in the Finnish Diabetes Prevention Study by Markus J. Takkunen; Ursula S. Schwab; Vanessa D. F. de Mello; Johan G. Eriksson; Jaana Lindström; Jaakko Tuomilehto; Matti I. J. Uusitupa (967-979).
To examine the longitudinal associations of serum fatty acid composition with type 2 diabetes, insulin secretion and insulin sensitivity over several years.We conducted a prospective cohort study derived from the randomized Finnish Diabetes Prevention Study. Total serum fatty acid composition was measured using gas chromatography in 407 overweight, middle-aged people with impaired glucose tolerance at baseline (1993–1998) and annually during the intervention period (1994–2000). Longitudinal associations of 20 fatty acids and three desaturase activities (Δ5 (20:4n-6/20:3n-6, D5D), Δ6 (18:3n-6/18:2n-6, D6D), stearoyl-CoA desaturase-1 (16:1n-7/16:0, SCD-1)) with type 2 diabetes incidence, and estimates of insulin sensitivity (Matsuda), secretion (ratio of insulin and glucose concentrations) and β-cell function (disposition index) by an oral glucose tolerance test were analyzed using Cox regression and linear mixed models. We validated estimated D5D and D6D using a known FADS1 gene variant, rs174550.The baseline proportions of 20:5n-3, 22:5n-3 and 22:6n-3, and D5D were associated with lower incidence of type 2 diabetes during a median follow-up of 11 years (HR per 1SD: 0.72, 0.74, 0.73, 0.78, respectively, P ≤ 0.01). These long-chain omega-3 fatty acids and D5D were associated with higher insulin sensitivity in subsequent years but not with disposition index. Saturated, monounsaturated and trans fatty acids and 18:3n-3, 18:2n-6, SCD-1 and D6D were inconsistently associated with type 2 diabetes or related traits.Serum long-chain omega-3 fatty acids and D5D predicted lower type 2 diabetes incidence in people at a high risk of diabetes attending to an intervention study; a putative mechanism behind these associations was higher insulin sensitivity.
Keywords: Type 2 diabetes; Biomarkers; Serum fatty acids; Fatty acid desaturases; Omega-3 fatty acids; Cohort study

Association between n-3 polyunsaturated fatty acids in erythrocytes and metabolic syndrome in Chinese men and women by Xiao-wei Dai; Yu-ming Chen; Fang-fang Zeng; Li-li Sun; Cao-gang Chen; Yi-xiang Su (981-989).
Evidence of an association between n-3 polyunsaturated fatty acids (PUFAs) and metabolic syndrome (MS) is limited and inconsistent. We investigated the association between n-3 PUFAs in erythrocytes and the presence of MS in Chinese adults.The levels of α-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA) in erythrocytes were measured using gas chromatography in 3072 participants (900 men and 2172 women) aged 30–75 years from Guangzhou, China. Cardiometabolic factors were determined, and MS was defined using the updated Adult Treatment Panel III criteria. Other covariates were collected via interviewer-administered questionnaires.After adjusting for age and other confounders, higher levels of marine-derived n-3 PUFAs, including EPA, DPA, and DHA, were associated with a lower presence of metabolic syndrome in both men and women. The odds ratios (95 % confidence interval) for MS obtained by comparing extreme quartiles were 0.55 (0.35–0.88) (EPA), 0.54 (0.34–0.87) (DPA), 0.45 (0.27–0.73) (DHA), and 0.52 (0.32–0.84) (total n-3 PUFAs) in men (p trend <0.05 for all results); and 0.74 (0.56–0.99) (EPA), 0.73 (0.55–0.98) (DPA), 0.75 (0.56–1.02) (DHA), and 0.71 (0.53–0.96) (total n-3 PUFAs) in women, respectively. No significant association of ALA with MS was observed (p trend > 0.05).Higher levels of total n-3 PUFAs, EPA, DPA, and DHA, but not of ALA, in erythrocyte membranes are associated with a lower presence of metabolic syndrome in Chinese adults.
Keywords: n-3 polyunsaturated fatty acids; Erythrocyte; Metabolic syndrome; Chinese adults

The New Nordic Diet: phosphorus content and absorption by Louise Salomo; Sanne K. Poulsen; Marianne Rix; Anne-Lise Kamper; Thomas M. Larsen; Arne Astrup (991-996).
High phosphorus content in the diet may have adverse effect on cardiovascular health. We investigated whether the New Nordic Diet (NND), based mainly on local, organic and less processed food and large amounts of fruit, vegetables, wholegrain and fish, versus an Average Danish Diet (ADD) would reduce the phosphorus load due to less phosphorus-containing food additives, animal protein and more plant-based proteins.Phosphorus and creatinine were measured in plasma and urine at baseline, week 12 and week 26 in 132 centrally obese subjects with normal renal function as part of a post hoc analysis of data acquired from a 26-week controlled trial. We used the fractional phosphorus excretion as a measurement of phosphorus absorption.Mean baseline fractional phosphorus excretion was 20.9 ± 6.6 % in the NND group (n = 82) and 20.8 ± 5.5 % in the ADD group (n = 50) and was decreased by 2.8 ± 5.1 and 3.1 ± 5.4 %, respectively, (p = 0.6) at week 26. At week 26, the mean change in plasma phosphorus was 0.04 ± 0.12 mmol/L in the NND group and −0.03 ± 0.13 mmol/L in the ADD group (p = 0.001). Mean baseline phosphorus intake was 1950 ± 16 mg/10 MJ in the NND group and 1968 ± 22 mg/10 MJ in the ADD group and decreased less in the NND compared to the ADD (67 ± 36 mg/10 MJ and −266 ± 45 mg/day, respectively, p < 0.298).Contrary to expectations, the NND had a high phosphorus intake and did not decrease the fractional phosphorus excretion compared with ADD. Further modifications of the diet are needed in order to make this food concept beneficial regarding phosphorus absorption.
Keywords: New Nordic Diet; Dietary phosphorus intake; Fractional phosphorus excretion; Plasma phosphorus; Urinary phosphorus excretion

Multiplatform metabolomic fingerprinting as a tool for understanding hypercholesterolemia in Wistar rats by Diana González-Peña; Danuta Dudzik; Clara Colina-Coca; Begoña de Ancos; Antonia García; Coral Barbas; Concepción Sánchez-Moreno (997-1010).
The aim was to investigate the impact of hypercholesterolemic diet on the metabolome of male Wistar rats by a multiplatform metabolomic fingerprinting.Male Wistar rats were fed with two different diets [control (C) and high-cholesterol diet (HC)—containing 2 % cholesterol and 0.5 % cholic acid]. After 7 weeks of experimental feeding, the rats were euthanized for blood collection and plasma recovery. The metabolite fingerprint was then achieved by applying a multiplatform comprising LC–MS, GC–MS and CE–MS.Multivariate statistical analysis showed a clear separation between the C and HC groups. Individual differences in metabolites were evaluated using univariate statistical analysis, and multiple metabolites were identified and confirmed in the plasma. A global profiling integrates for the first time pathways affected by high-cholesterol diet intake and allowed us to elucidate some of the associated alterations underlying the hypercholesterolemia event in Wistar rats.HC feeding stimulated the alteration of multiple pathways in Wistar rats, warning of the risk of developing important diseases, which can be modulated by the diet. Further studies are required to investigate the possibilities to revert or ameliorate the negative effects triggered by HC intake.
Keywords: Atherosclerosis; Global profiling; High-cholesterol diet; Metabolomics; Untargeted analysis

There is limited evidence whether environmental exposure to perfluorinated compounds (PFCs) affects insulin resistance (IR) and whether vitamin C intake protects against the adverse effect of PFCs. This study was carried out to investigate the effect of PFCs on IR through oxidative stress, and the effects of a 4-week consumption of vitamin C supplement compared placebo on development of IR by PFCs.For a double-blind, community-based, randomized, placebo-controlled crossover intervention of vitamin C, we assigned 141 elderly subjects to both vitamin C and placebo treatments for 4 weeks. We measured serum levels of PFCs to estimate PFC exposures and urinary levels of malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) for oxidative stress. We also measured levels of fasting glucose and insulin and derived the homeostatic model assessment (HOMA) index to assess IR.Perfluorooctane sulfonate (PFOS) and perfluorododecanoic acid (PFDoDA) levels were found to be positively associated with HOMA index at the baseline and after placebo treatment. Risks of IR for the top decile of PFOS and PFDoDA exposures were significantly elevated compared with those with lower PFOS and PFDoDA exposures (both, P < 0.0001). However, the effects of PFOS and PFDoDA on HOMA disappeared after vitamin C supplementation (both, P > 0.30). Furthermore, PFOS and PFDoDA levels were also significantly associated with MDA and 8-OHdG levels, and MDA levels were positively associated with HOMA index.PFOS and PFDoDA exposures were positively associated with IR and oxidative stress, and vitamin C supplementation protected against the adverse effects of PFOS and PFDoDA on IR.
Keywords: Perfluorinated compounds; Insulin resistance; Oxidative stress; Vitamin C supplementation

Folic acid causes higher prevalence of detectable unmetabolized folic acid in serum than B-complex: a randomized trial by Rima Obeid; Susanne H. Kirsch; Sarah Dilmann; Cosima Klein; Rudolf Eckert; Jürgen Geisel; Wolfgang Herrmann (1021-1028).
Unmetabolized folic acid (UMFA) is common in serum of elderly individuals receiving folic acid (FA)-fortified foods or supplements. We studied the effect of supplementing FA or B-complex on serum concentrations of (6S)-5-methyltetrahydropteroylglutamate [(6S)-5-CH3-H4Pte] and UMFA in elderly people and explored factors associated with detectable UMFA post-supplementation.This is a randomized single-blind non-controlled trial on 58 elderly people using daily 400 µg FA (n = 31) or 400 µg FA, 10 µg cyanocob(III)alamin and 8 mg pyridoxine (n = 27) for a median of 23 days. Main outcome includes changes in concentrations of serum (6S)-5-CH3-H4Pte and UMFA.Total homocysteine declined by a median of 1.6 (p = 0.074) in the FA and 1.3 µmol/L (p = 0.009) in the B-complex arms (p = 0.66 between the arms). Serum (6S)-5-CH3-H4Pte significantly (p < 0.001 vs. baseline) increased by a median of 9.2 and 6.5 nmol/L in the FA and B-complex groups, respectively (p = 0.152 between the groups). Compared to FA, B-complex reduced cystathionine and caused lower post-intervention serum UMFA, percentage of UMFA to (6S)-5-CH3-H4Pte and prevalence of UMFA ≥ 0.21 nmol/L. Higher serum cystathionine and whole-blood folate predicted higher post-intervention serum UMFA.FA caused higher UMFA as compared to B-complex. Pyridoxine appears to improve folate recycling. Data on serum UMFA should be interpreted in relation to other vitamins involved in folate metabolism. Serum UMFA is suggested to play a sensory role through which the cell recognizes FA available for metabolism via dihydrofolate reductase.
Keywords: Unmetabolized folic acid; Elderly; Supplementation; Vitamin B6; Vitamin B12; Serum folate

Urinary phytoestrogens and cancer, cardiovascular, and all-cause mortality in the continuous National Health and Nutrition Examination Survey by Michael K. Reger; Terrell W. Zollinger; Ziyue Liu; Josette Jones; Jianjun Zhang (1029-1040).
Experimental studies suggest that phytoestrogen intake alters cancer and cardiovascular risk. This study investigated the associations of urinary phytoestrogens with total cancer (n = 79), cardiovascular (n = 108), and all-cause (n = 290) mortality among 5179 participants in the continuous National Health and Nutrition Examination Survey (1999–2004).Urinary phytoestrogens were measured using high-performance liquid chromatography with tandem mass spectrometric detection. Survival analysis was performed to evaluate hazard ratios (HRs) and 95 % confidence intervals (CIs) for each of the three outcomes in relation to urinary phytoestrogens.After adjustment for confounders, higher urinary concentrations of total enterolignans were associated with a reduced risk of death from cardiovascular disease (HR for tertile 3 vs. tertile 1 0.48; 95 % CI 0.24, 0.97), whereas higher urinary concentrations of total isoflavones (HR for tertile 3 vs. tertile 1 2.14; 95 % CI 1.03, 4.47) and daidzein (HR for tertile 3 vs. tertile 1 2.05; 95 % CI 1.02, 4.11) were associated with an increased risk. A reduction in all-cause mortality was observed for elevated urinary concentrations of total enterolignans (HR for tertile 3 vs. tertile 1 0.65; 95 % CI 0.43, 0.96) and enterolactone (HR for tertile 3 vs. tertile 1 0.65; 95 % CI 0.44, 0.97).Some urinary phytoestrogens were associated with cardiovascular and all-cause mortality in a representative sample of the US population. This is one of the first studies that used urinary phytoestrogens as biomarkers of their dietary intake to evaluate the effect of these bioactive compounds on the risk of death from cancer and cardiovascular disease.
Keywords: Cancer; Cardiovascular disease; Cohort study; Mortality; Urinary phytoestrogens

Modulation of cholesterol-related gene expression by ergosterol and ergosterol-enriched extracts obtained from Agaricus bisporus by Alicia Gil-Ramírez; Víctor Caz; Roberto Martin-Hernandez; Francisco R. Marín; Carlota Largo; Arantxa Rodríguez-Casado; María Tabernero; Alejandro Ruiz-Rodríguez; Guillermo Reglero; Cristina Soler-Rivas (1041-1057).
To investigate the effect of two extracts obtained from Agaricus bisporus on the mRNA expression of cholesterol-related genes. One of the extracts contained ergosterol and other fungal sterols (SFE) and the other contained β-glucans and fungal sterols (EβG).Firstly, the dietary mixed micelles (DMMs) generated after in vitro digestion of standards and SFE were applied to Caco2 cells. Then, the lower compartment after a Caco2-transport experiment was applied to HepG2 cells. The mRNA expression was assessed in both cell lines by low-density arrays (LDA). Mice received the extracts, ergosterol or control drugs after 4 weeks of a high-cholesterol diet. The lipid profile of plasma, liver and feces was determined. LDA assays were performed in liver and intestines.The DMM fraction of SFE up-regulated the LDLR mRNA expression in Caco2 cells. The lower compartment after Caco2-transport experiments up-regulated LDLR and modulated several other lipid-related genes in HepG2 cells. In mice, SFE decreased TC/HDL ratio and reduced hepatic triglycerides paralleled with down-regulation of Dgat1 expression, while EβG did it without transcriptional changes. Addition of SFE or ergosterol induced in jejunum a similar transcriptional response to simvastatin and ezetimibe; they all down-regulated Srebf2 and Nr1h4 (FXR) genes.Ergosterol-containing extracts from A. bisporus lowered hepatic triglyceride and modify the mRNA expression of cholesterol-related genes although the transcriptional regulation was unrelated to changes in plasma lipid profile. These extracts may be useful limiting hepatic steatosis and as bioactive ingredients to design novel functional foods preventing lifestyle-related diseases such as non-alcoholic fatty liver disease.
Keywords: Ergosterol; Cholesterol; Supercritical CO2 extraction; White button mushroom; Low-density array (LDA); Gene expression

Recent studies suggest that nutritional status during developmental periods is associated with subsequent development of metabolic abnormalities. In this study, we examined whether malnutrition by fasting for 3 days during the suckling–weaning transient period induces subsequent development of metabolic abnormalities in rats.Male Sprague–Dawley rats were fasted for 3 days during the suckling–weaning transient period. They are subsequently fed a high-fat, high-sucrose (HF) or low-fat, high-starch (LF) diet for 14 weeks from 17 weeks of age, and the liver and blood samples were collected for measuring mRNA and protein levels of metabolic genes and blood concentrations of glucose and insulin, respectively.Fasting for 3 days during the suckling–weaning transient period induced impaired glucose tolerance in rats fed the LF diet in adulthood. Liver triglycerides in rats fed the HF diet in adulthood increased to 140 % in rats fasted for 3 days during the suckling–weaning transient period compared with those non-fasted. Furthermore, liver expression of FBP1 and ACCα genes in adult rats fed the LF diet increased to 125 and 145 %, respectively, in rats fasted for 3 days during the suckling–weaning transient period compared to non-fasted rats. PEPCK1 protein expression levels in rats fed the LF diet were higher in rats fasted for 3 days during the suckling–weaning transient period than in non-fasted rats.Fasting for 3 days in rats during the suckling–weaning transient period enhances metabolic abnormalities in animals fed a HF or LF diet in adulthood by confounding metabolism of lipid and sugar in the liver.
Keywords: Metabolic abnormalities; Malnutrition; Suckling–weaning transient period; Impaired glucose tolerance

Dietary inflammatory index and risk of lung cancer and other respiratory conditions among heavy smokers in the COSMOS screening study by Patrick Maisonneuve; Nitin Shivappa; James R. Hébert; Massimo Bellomi; Cristiano Rampinelli; Raffaella Bertolotti; Lorenzo Spaggiari; Domenico Palli; Giulia Veronesi; Patrizia Gnagnarella (1069-1079).
To test whether the inflammatory potential of diet, as measured using the dietary inflammatory index (DII), is associated with risk of lung cancer or other respiratory conditions and to compare results obtained with those based on the aMED score, an established dietary index that measures adherence to the traditional Mediterranean diet.In 4336 heavy smokers enrolled in a prospective, non-randomized lung cancer screening program, we measured participants’ diets at baseline using a self-administered food frequency questionnaire from which dietary scores were calculated. Cox proportional hazards and logistic regression models were used to assess association between the dietary indices and lung cancer diagnosed during annual screening, and other respiratory outcomes that were recorded at baseline, respectively.In multivariable analysis, adjusted for baseline lung cancer risk (estimated from age, sex, smoking history, and asbestos exposure) and total energy, both DII and aMED scores were associated with dyspnoea (p trend = 0.046 and 0.02, respectively) and radiological evidence of emphysema (p trend = 0.0002 and 0.02). After mutual adjustment of the two dietary scores, only the association between DII and radiological evidence of emphysema (Q4 vs. Q1, OR 1.30, 95 % CI 1.01–1.67, p trend = 0.012) remained statistically significant. At univariate analysis, both DII and aMED were associated with lung cancer risk, but in fully adjusted multivariate analysis, only the association with aMED remained statistically significant (p trend = 0.04).Among heavy smokers, a pro-inflammatory diet, as indicated by increasing DII score, is associated with dyspnoea and radiological evidence of emphysema. A traditional Mediterranean diet, which is associated with a lower DII, may lower lung cancer risk.
Keywords: Dietary inflammatory index; Mediterranean diet; Lung cancer; Lung function; Emphysema; Dyspnoea

Olfactory impairment in older adults is associated with poorer diet quality over 5 years by Bamini Gopinath; Joanna Russell; Carolyn M. Sue; Victoria M. Flood; George Burlutsky; Paul Mitchell (1081-1087).
Decreased smell could cause appetite suppression and malnutrition. However, there is a paucity of longitudinal data between olfaction and nutritional status in older adults. We aimed to prospectively examine the relationship between olfactory impairment and overall diet quality (reflecting adherence to dietary guidelines) in a population-based cohort of older adults.We used 5-year follow-up data from 557 adults (aged 60+ years at baseline) whose olfaction was measured using the San Diego Odor Identification Test (SDOIT). Dietary data were collected using a validated semiquantitative food frequency questionnaire. A total diet score (TDS) was calculated for intake of selected food groups and nutrients for each participant as described in the national dietary guidelines. Final scores ranged from 0 to 20; higher scores indicated closer adherence to dietary guidelines.After adjusting for all potential confounders, older adults with moderate/severe olfactory impairment (SDOIT score ≤ 3; lower scores indicate impairment) compared with those with no olfactory impairment had significantly lower adjusted mean (±SE) TDS, 9.09 (0.40) versus 9.94 (0.10), p = 0.04. Women with moderate/severe impaired olfaction (i.e., scored poorly on the odor identification test) compared with those with normal olfaction had significantly lower adjusted mean TDS, 8.87 (0.69) versus 10.31 (0.13), p = 0.04. No associations were observed between olfaction and TDS in men.Olfactory impairment in older women could signal an increased risk of poorer diet quality, defined as adherence to national dietary guidelines. Additional longitudinal studies are needed to confirm or refute the observed link between olfactory loss and overall patterns of food intake in older adults.
Keywords: Olfactory impairment; Diet quality; Older adults; Blue Mountains Eye Study

Association between obesity and glomerular hyperfiltration: the confounding effect of smoking and sodium and protein intakes by Adam Ogna; Valentina Forni Ogna; Murielle Bochud; Idris Guessous; Fred Paccaud; Michel Burnier; Gregoire Wuerzner (1089-1097).
Glomerular hyperfiltration has been suggested as a possible mechanism linking obesity and chronic kidney disease (CKD), independently of classical risk factors. We explored the association of overweight and obesity with glomerular hyperfiltration in a large sample of the Swiss adult population, accounting for several confounders including dietary factors.Data from a 2010 to 2012 cross-sectional population-based survey in Switzerland were used. Creatinine clearance (CrCl) was determined from 24-h urine collection; CrCl > 140 ml/min was used to define glomerular hyperfiltration. Participants were categorized into lean (<25 kg/m2), overweight (25–29.9 kg/m2) and obese (≥30 kg/m2) according to body mass index (BMI).A total of 1339 participants were included in the analysis [median (IQR) age 49.4 (34.3–63.5) years, 48.9 % men]. The prevalences of overweight and obesity were 32.2 and 14.2 %, respectively. Median CrCl was 102[84–121] ml/min in lean, 110 [87–136] ml/min in overweight and 124 [97–150] ml/min in obese participants (p < 0.001). The prevalence of glomerular hyperfiltration increased across BMI categories (10.4, 20.8 and 34.7 %, respectively; p < 0.001). This positive association remained significant after adjusting for age, sex, hypertension, diabetes, smoking and dietary factors (sodium and protein intakes): odds ratio [95 %CI] 2.39 [1.52–3.76] (p < 0.001) for overweight versus lean and 4.10[2.31–7.27] (p < 0.001) for obesity versus lean.BMI categories and glomerular hyperfiltration are positively associated, independently of other known CKD risk factors and dietary confounders, suggesting that glomerular hyperfiltration may represent an early renal phenotype in obesity. Our observations confirm the significant association of glomerular hyperfiltration with sodium and protein intakes and identify sodium intake as an important modifying factor of the association between hyperfiltration and obesity.
Keywords: Obesity; Overweight; Kidney disease; Glomerular hyperfiltration; Dietary factors

Urolithin A causes p21 up-regulation in prostate cancer cells by Claudia Sánchez-González; Carlos J. Ciudad; Maria Izquierdo-Pulido; Véronique Noé (1099-1112).
Walnuts contain several bioactive compounds, including pedunculagin, a polyphenol metabolized by microbiota to form urolithins, namely urolithin A (UA). The aim of this study was to determine gene expression changes in prostate cancer cells after incubation with UA.We performed a genomic analysis to study the effect of UA on LNCaP prostate cells. Cells were incubated with 40 µM UA for 24 h, and RNA was extracted and hybridized to Affymetrix Human Genome U219 array. Microarray results were analyzed using GeneSpring v13 software. Differentially expressed genes (p < 0.05, fold change > 2) were used to perform biological association networks. Cell cycle was analyzed by flow cytometry and apoptosis measured by the rhodamine method and by caspases 3 and 7 activation. Cell viability was determined by MTT assay.We identified two nodes, FN-1 and CDKN1A, among the differentially expressed genes upon UA treatment. CDKN1A was validated, its mRNA and protein levels were significantly up-regulated, and the promoter activation measured by luciferase. Cell cycle analysis showed an increase in G1-phase, and we also observed an induction of apoptosis and caspases 3 and 7 activation upon UA treatment.Our results indicate a potential role of UA as a chemopreventive agent for prostate cancer.
Keywords: Urolithin A; Walnuts; Ellagitannins cancer prevention

Moderate physical training attenuates perinatal low-protein-induced spleen lymphocyte apoptosis in endotoxemic adult offspring rats by Sueli Moreno Senna; Marília Kalinne Torres; Daíllo Augusto Pereira Lopes; Maria Claudia Alheiros-Lira; Diógenis Barbosa de Moura; Valéria Rêgo Alves Pereira; Francisco Carlos Amanajás de Aguiar Jr.; José Candido Ferraz; Carol Góis Leandro (1113-1122).
To evaluate the effects of a moderate physical training (T) on the blood and splenic lymphocytes subsets and the rate of apoptosis in adult offspring submitted to perinatal low-protein (LP) diet.Male Wistar rats were divided according to their mother’s diet: control (C, 17 % casein) and undernourished (LP, 8 % casein). At the 60th day, pups were submitted to moderate physical training (8 weeks, 5 days week−1, 60 min day−1, at 70 % of VO2max). After T period, pups received an injection of lipopolysaccharide (LPS). B, NK, and TCD3+ lymphocytes subsets were analyzed by flow cytometry. Spleen lymphocytes apoptosis was evaluated by DNA fragmentation, phosphatidylserine externalization (PSE), and mitochondrial transmembrane depolarization (MTD) using a flow cytometer. Plasma TNF-α concentrations were analyzed by ELISA.LP + LPS pups showed a higher percentage of blood B, CD4+, and NK and a reduction in TCD3+, CD8+ than C pups. The percentage of NK and CD3+ was restored in LP + T + LPS pups. In the spleen, T normalized the percentage of NK in LP + LPS pups. LP + LPS pups showed a higher percentage of cells with PSE and MTD than C + LPS pups that was attenuated by T. The concentration of TNF-α was higher in LP + LPS than C + LPS, but it was attenuated in LP + T + LPS pups.Moderate physical training was able to revert the effects of perinatal LP diet on circulation lymphocytes subsets and attenuated splenic lymphocytes apoptosis and plasma TNF-α concentrations.
Keywords: Protein malnutrition; Developmental plasticity; Rats; Physical exercise; Splenic lymphocytes; Cell death, LPS

This randomised controlled trial assessed the acute and long-term effects of daily supplementation of kanuka honey, formulated with cinnamon, chromium and magnesium on glucose metabolism, weight and lipid parameters in individuals with type 2 diabetes. Twelve individuals with type 2 diabetes received 53.5 g of a formulated honey and a control (non-formulated) kanuka honey in a random order for 40 days, using cross-over design. Fasting glucose, insulin, HbA1c, lipids and anthropometric measures were measured at baseline and end of treatment. A meal tolerance test was performed at baseline to assess acute metabolic response.There was no statistically significant difference in acute glucose metabolism between treatment groups, as measured by the Matsuda index and AUC for glucose and insulin. After the 40-day intervention with honey, fasting glucose did not differ significantly between the two treatments (95 % CI −2.6 to 0.07). There was no statistically significant change in HbA1c or fasting insulin. There was a statistically significant reduction in total cholesterol by −0.29 mmol/L (95 % CI −0.57 to −0.23), LDL cholesterol by −0.29 mmol/L (95 % CI −0.57 to −0.23) and weight by −2.2 kg (95 % CI −4.2 to −0.1). There was a trend towards increased HDL and reduced systolic blood pressure in the intervention treatment.The addition of cinnamon, chromium and magnesium supplementation to kanuka honey was not associated with a significant improvement in glucose metabolism or glycaemic control in individuals with type 2 diabetes. Use of the formulated honey was associated with a reduction in weight and improvements in lipid parameters, and should be investigated further.
Keywords: Diabetes; Honey; Cinnamon; Chromium; Magnesium; Lipids

Adherence to the Mediterranean diet and academic performance in youth: the UP&DOWN study by Irene Esteban-Cornejo; Rocio Izquierdo-Gomez; Sonia Gómez-Martínez; Carmen Padilla-Moledo; Jose Castro-Piñero; Ascensión Marcos; Oscar L. Veiga (1133-1140).
To examine the association between adherence to the Mediterranean diet and academic performance in children and adolescents.This is a cross-sectional study conducted with 1371 youth aged 12.04 ± 2.50 years (685 girls) in Spain during 2011–2012. Adherence to the Mediterranean diet was assessed using the KIDMED index (Mediterranean Diet Quality Index in children and adolescents), which includes 16 questions on specific dietary patterns. Levels of adherence were classified into three groups: poor adherence (0–3), average adherence (4–7), and good adherence (8–12). Academic performance was assessed through school records using four indicators: math, language, an average of math and language, and grade point average score.Adherence to the Mediterranean diet was related to academic performance (β ranging from 0.107 to 0.148; all P < 0.001) after adjusting for confounders. The group of good adherence to the Mediterranean diet had significantly higher scores in all of the academic indicators compared with the poor group (ranging from +0.429 to 0.464; all P ≤ 0.001); as well as the group of average adherence to the Mediterranean diet had significantly higher scores in all of the academic indicators compared with the poor group (ranging from +0.292 to 0.344; all P ≤ 0.06). There were no differences between the groups of good and average adherence to the Mediterranean diet.Adherence to the Mediterranean diet may have a beneficial influence on academic performance in youth. Importantly, the benefits of adherence to the Mediterranean diet on academic performance may be stronger as youth adhered to the optimal Mediterranean diet levels.
Keywords: Mediterranean diet; Academic performance; Children and adolescents

The development of allergic inflammation in a murine house dust mite asthma model is suppressed by synbiotic mixtures of non-digestible oligosaccharides and Bifidobacterium breve M-16V by K. A. T. Verheijden; L. E. M. Willemsen; S. Braber; T. Leusink-Muis; P. V. Jeurink; J. Garssen; A. D. Kraneveld; G. Folkerts (1141-1151).
The incidence and severity of allergic asthma is rising, and novel strategies to prevent or treat this disease are needed. This study investigated the effects of different mixtures of non-digestible oligosaccharides combined with Bifidobacterium breve M-16V (BB) on the development of allergic airway inflammation in an animal model for house dust mite (HDM)-induced allergic asthma.BALB/c mice were sensitized intranasally (i.n.) with HDM and subsequently challenged (i.n.) with PBS or HDM while being fed diets containing different oligosaccharide mixtures in combination with BB or an isocaloric identical control diet. Bronchoalveolar lavage fluid (BALF) inflammatory cell influx, chemokine and cytokine concentrations in lung homogenates and supernatants of ex vivo HDM-restimulated lung cells were analyzed.The HDM-induced influx of eosinophils and lymphocytes was reduced by the diet containing the short-chain and long-chain fructo-oligosaccharides and BB (FFBB). In addition to the HDM-induced cell influx, concentrations of IL-33, CCL17, CCL22, IL-6, IL-13 and IL-5 were increased in supernatants of lung homogenates or BALF and IL-4, IFN-γ and IL-10 were increased in restimulated lung cell suspensions of HDM-allergic mice. The diet containing FFBB reduced IL-6, IFN-γ, IL-4 and IL-10 concentrations, whereas the combination of galacto-oligosaccharides and long-chain fructo-oligosaccharides with BB was less potent in this model.These findings show that synbiotic dietary supplementation can affect respiratory allergic inflammation induced by HDM. The combination of FFBB was most effective in the prevention of HDM-induced airway inflammation in mice.
Keywords: House dust mite; Asthma; Allergy; Oligosaccharides; Bifidobacterium breve

Moderate alcohol consumption diminishes the development of non-alcoholic fatty liver disease (NAFLD) in ob/ob mice by Giridhar Kanuri; Marianne Landmann; Josephine Priebs; Astrid Spruss; Marina Löscher; Doreen Ziegenhardt; Carolin Röhl; Christian Degen; Ina Bergheim (1153-1164).
Using ob/ob mice as a model of non-alcoholic fatty liver disease (NAFLD), we investigated the effect of moderate alcohol intake on the development of NAFLD and molecular mechanisms involved. Ob/ob mice were fed water or ethanol solution (2.5 g/kg body weight/day) for 6 weeks, and markers of liver injury, insulin signalling and adiponectin in visceral adipose tissue were determined.Whereas bodyweight and the degree of liver steatosis did not differ among ob/ob mouse groups, those consuming ethanol had markedly less macrovesicular hepatic fat accumulation, inflammatory alterations and significantly lower transaminase levels. Despite similarly elevated protein levels of tumour necrosis factor α, protein concentrations of plasminogen activator inhibitor 1 were significantly lower in livers of ob/ob mice consuming ethanol in comparison with controls. The hepato-protective property of moderate alcohol ingestion in ob/ob mice was associated with an induction of the sirtuin-1/adiponectin-signalling cascade in visceral fat tissue and an activation of Akt in the liver. Similar effects of moderate alcohol exposure were also found in vitro in 3T3-L1 and AML-12 cells.These data suggest that moderate alcohol intake may diminish the development of NAFLD through sirtuin-1/-adiponectin-dependent signalling cascades.
Keywords: Adiponectin; Ethanol; PAI-1; SIRT1; Visceral fat

Tamoxifen has been used for the treatment of estrogen receptor (ER)-positive breast cancers and in women who are at an increased risk of breast cancer. Acquired resistance to this drug and its toxicity still pose a clinically significant problem, especially in the prevention setting. Isothiocyanates present in cruciferous plants, such as sulforaphane or erucin, have been shown to reduce growth of breast cancer cells in vivo and in vitro. In this study, we explored their ability to sensitize cancer cells to 4-hydroxytamoxifen. We used three ER-positive breast cancer cell lines, T47D, MCF-7 and BT-474, as well as the drug-resistant T47D and MCF-7 derivatives. We examined the effect of 4-hydroxytamoxifen, isothiocyanates and their combinations on cell viability by MTT and clonogenic assays. Impact of treatments on the levels of proteins engaged in apoptosis and autophagy was determined by Western blotting.Isothiocyanates act in a synergistic way with 4-hydroxytamoxifen, and co-treatment reduces breast cancer cell viability and clonogenic potential more effectively than treatment with any single agent. This is connected with a drop in the Bcl-2/Bax ratio and the level of survivin as well as increased PARP cleavage, and elevation in ADRP, the mitochondrial stress marker. Moreover, isothiocyanates sensitize 4-hydroxytamoxifen-resistant T47D and MCF-7 cells to the drug.Isothiocyanates enhance response to 4-hydroxytamoxifen, which allows for reduction of the effective drug concentration. Combinatorial strategy may hold promise in development of therapies and chemoprevention strategies against ER-positive breast tumors, even those with acquired resistance to the drug.
Keywords: Breast cancer; Sulforaphane; Erucin; 4-Hydroxytamoxifen; Estrogen receptor

Changes in macronutrient, micronutrient, and food group intakes throughout the menstrual cycle in healthy, premenopausal women by Anna M. Gorczyca; Lindsey A. Sjaarda; Emily M. Mitchell; Neil J. Perkins; Karen C. Schliep; Jean Wactawski-Wende; Sunni L. Mumford (1181-1188).
It is thought that total energy intake in women is increased during the luteal versus follicular phase of the menstrual cycle; however, less is understood regarding changes in diet composition (i.e., macro- and micronutrient intakes) across the cycle. The aim of this study was to investigate changes in macronutrient, micronutrient, and food group intakes across phases of the menstrual cycle among healthy women, and to assess whether these patterns differ by ovulatory status.The BioCycle study (2005–2007) was a prospective cohort study of 259 healthy regularly menstruating women age 18–44 who were followed for up to two menstrual cycles. Dietary intake was measured using 24-h dietary recalls, and food cravings were assessed via questionnaire, up to four times per cycle, corresponding to menses, mid-follicular, expected ovulation, and luteal phases. Linear mixed models adjusting for total energy intake were used to evaluate changes across the cycle.Total protein (P = 0.03), animal protein (P = 0.05), and percent of caloric intake from protein (P = 0.02) were highest during the mid-luteal phase compared to the peri-ovulatory phase. There were also significant increases in appetite, craving for chocolate, craving for sweets in general, craving for salty flavor, and total craving score during the late luteal phase compared to the menstrual, follicular, and ovulatory phases (P < 0.001).Our findings suggest an increased intake of protein, and specifically animal protein, as well as an increase in reported food cravings, during the luteal phase of the menstrual cycle independent of ovulatory status. These results highlight a plausible link between macronutrient intake and menstrual cycle phase.
Keywords: Menstrual cycle; Macronutrients; Micronutrients; Anovulatory

NMR-based metabolomics highlights differences in plasma metabolites in pigs exhibiting diet-induced differences in adiposity by Maëva Jégou; Florence Gondret; Julie Lalande-Martin; Illa Tea; Elisabeth Baéza; Isabelle Louveau (1189-1199).
A better understanding of the control of body fat mass and distribution is required for both human health and animal production. The current study investigates plasma parameters in response to changes in body fat mass.Pigs from two lines divergently selected for residual feed intake were fed diets contrasted in energy sources and nutrients. Between 74 and 132 days of age, pigs (n = 12 by diet and by line) received isoproteic and isoenergetic diets, either rich in starch (LF) or in lipids and fibres (HF). At the end of the feeding trial, plasma samples were analysed by 1H NMR spectroscopy and standard hormonal and biochemical assays.Pigs fed the HF diet had lower (P < 0.01) perirenal and subcutaneous adipose tissue relative masses than pigs fed the LF diet. Metabolomic approach showed a clear discrimination between diets, with lower (P < 0.05) plasma levels of creatinine–lysine, creatine, tyrosine, proline, histidine, lysine, phenylalanine and formate but higher (P < 0.001) plasma VLDL-LDL levels in HF pigs than in LF pigs. Plasma concentrations of triglycerides were higher (P < 0.001), while plasma concentrations of β-hydroxybutyrate, leptin, glucose, insulin and urea were lower (P ≤ 0.05) in HF pigs than in LF pigs. Plasma levels of leptin, creatine and urea were positively correlated (r = 0.3, P < 0.05) with relative adipose tissue masses.These data indicate that metabolites associated with energy and protein metabolism were involved in the response to a high-fat, high-fibre diet. Relevant plasma indicators of metabolic flexibility related to changes in body adiposity were then proposed.
Keywords: Amino acids; Fatness; High-fibre diet; Leptin; NMR spectroscopy

To measure the iodine status and iodine intake of New Zealand adults 18–64 years of age following mandatory fortification of bread with iodine.A cross-sectional survey of NZ adults living in Dunedin and Wellington during February–November 2012. Three hundred and one men and women aged 18–64 years randomly selected from the New Zealand Electoral Roll completed a 24-h urine collection, a demographic and iodine-specific food frequency questionnaire (FFQ), and had height and weight measured. Urine collections were analysed for iodine and reported as median urinary iodine concentration (UIC) µg/L and median urinary iodine excretion (UIE) µg/day. The FFQ was used to estimate iodine intake with and without discretionary iodised salt use.The median UIC for all adults was 73 µg/L, indicative of mild iodine deficiency. The mean urinary volume was 2.0 L. As an estimate of iodine intake, the median UIE was 127 µg/day. Estimated iodine intake, using the FFQ which included discretionary iodised salt use, was 132 µg/day. Iodine intakes were associated with UIC (P = 0.040) and UIE (P = 0.003), but not with bread iodine intake and iodised salt use.Using the WHO/UNICEF/ICCIDD target for iodine sufficiency (a UIC of >100 µg/L) based on school-aged children with a mean urinary volume of 1.0 L, the iodine status of NZ adults does not reach adequate levels (73 µg/L). A more realistic parameter in a population with a higher urinary volume excretion (2.0 L) is the UIE. A median UIE of 127 µg/day suggests that the iodine status of NZ adults is now likely to be adequate.
Keywords: Iodine; Urinary iodine; Iodised salt; Fortification; Bread

Vitamin B6 status in the body is affected by several factors including dietary supply of the antivitamin B6 factor, 1-amino d-proline (1ADP), which is present in flaxseed. Owing to the prevalence of moderate B6 deficiency in the general population, a co-occurrence of 1ADP may lead to a further deterioration of B6 status. To this end, we applied a nontargeted metabolomics approach to identify potential plasma lipophilic biomarkers of deleterious effect of 1ADP on moderately vitamin B6-deficient rats using a high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.Twenty-four rats were fed with a semi-purified diet containing pyridoxine·HCl (PN·HCl) either 7 mg/kg diet (optimal B6) or 0.7 mg/kg diet (moderate B6). The rats were divided into four treatments (n = 6), and one treatment in each B6 diet group was also fed ad libitum with 10 mg/kg diet of synthetic 1ADP. After 5 weeks of study, plasma was collected from the rats and lipophilic metabolites were extracted using acetonitrile as a solvent for analysis. Ten potential plasma lipophilic biomarkers were identified out of >2500 detected entities, which showed significant differences between the treatments. Plasma glycocholic acid, glycoursodeoxycholic acid, murocholic acid, N-docosahexaenoyl GABA, N-arachidonoyl GABA, lumula, nandrolone and orthothymotinic acid concentrations were significantly elevated, while plasma cystamine and 3-methyleneoxindole concentrations were significantly reduced as a result of either low B6 status or 1ADP or their interaction.Changes in these metabolites revealed a potential defect in pathways linked with the biosynthesis and metabolism of bile acid components, N-acyl amino acids, analgesic androgens, anti-inflammatory and neuroprotective molecules. We also noted that the changes in these biomarkers can be alleviated by the application of adequate vitamin B6.
Keywords: Metabolomics; Biomarkers; Vitamin B6 ; Antivitamin B6 ; 1-Amino d-proline

Do bread-crust-derived Maillard reaction products affect the retention and tissue distribution of trace elements? by Cristina Delgado-Andrade; Irene Roncero-Ramos; Ana Haro; Silvia Pastoriza; María Pilar Navarro (1225-1233).
To investigate the effects of the consumption of Maillard reaction products (MRPs) from bread crust (BC) on iron, copper and zinc body retention and tissue distribution, determining whether these effects are related to the molecular weight of browning products. During an 88-day study period, rats were fed a Control diet or diets containing BC as source of MRPs, its soluble high or low molecular weight fractions (BC, LMW or HMW diets). A mineral balance was conducted throughout the experiment to determine iron, copper and zinc retention. At day 88, animals were killed, blood was drawn for haemoglobin determination and some organs removed to analyse minerals. Copper and zinc balances were unchanged, and scant modification detected in their body delivery. However, the Fe retention rate from the diet increased (13, 22 and 32 % for BC, LMW and HMW diets), and a parallel higher Fe body concentration was observed (13–18 % higher than the Control group). Incoming iron accumulated particularly in the liver, femur and small intestine, but functional iron tended to decrease, as reflected by haemoglobin levels.The long-term intake of BC or derivatives did not produce a notable effect on copper or zinc balances, although slightly increased iron retention rate and the body concentration of this mineral were observed. Iron accumulated in some organs, but the production of haemoglobin was not improved. In view of the differences observed between the effects of BC and its derivatives, our results underline the importance of working with real food matrices, where the joint presence of different components modulates the in vivo final effects.
Keywords: Maillard reaction products; Iron; Copper; Zinc; Retention; Tissue distribution

Comparison of plasma alkylresorcinols (AR) and urinary AR metabolites as biomarkers of compliance in a short-term, whole-grain intervention study by Nicola M. McKeown; Matti Marklund; Jiantao Ma; Alastair B. Ross; Alice H. Lichtenstein; Kara A. Livingston; Paul F. Jacques; Helen M. Rasmussen; Jeffrey B. Blumberg; C.-Y. Oliver Chen (1235-1244).
Alkylresorcinols (AR) are phenolic lipids present in the bran of wheat and rye. Plasma AR and their urinary metabolites may be suitable biomarkers of whole-grain (WG) wheat and rye consumption. The objective of this study was to examine plasma AR and urinary AR metabolites in response to WG wheat consumption.In a randomized crossover study, 19 subjects (10 males, 9 females; BMI 22.0 kg/m2; age 26 years) incorporated either 3 servings (48 g) or 6 servings (96 g) of WG wheat daily into their regular diet for 1 week. Subjects completed a 2-week washout period, abstaining from all WG consumption, before each intervention. Fasting blood and 24-h urine were collected before and after each intervention. Plasma AR homologues (C19:0, C21:0, C23:0) were quantified by GC–MS after diethyl ether and solid phase extraction and derivatization. Urinary AR metabolites [3,5-dihydroxybenzoic acid and 3-(3,5-dihydroxyphenyl)-propanoic acid] were determined using HPLC with electrochemical detection after enzymatic deconjugation and ethyl acetate extraction.Urinary total AR metabolites were significantly higher after 6 compared with 3 servings of WG wheat (56 vs. 32 μmol/day, P < 0.001). This dose–response relationship was independent of age, sex, energy intake, and baseline urinary AR metabolite concentration. Plasma total AR tended to be higher after 6 compared with 3 servings of WG wheat (103.0 vs. 86.9 nmol/L), but this difference was not significant (P = 0.42).The results suggest that urinary AR metabolites from 24-h urine collections may be useful as biomarkers of compliance in intervention studies of WG wheat.
Keywords: Alkylresorcinols; Biomarkers; Whole grain; Whole wheat

Vinegar decreases blood pressure by down-regulating AT1R expression via the AMPK/PGC-1α/PPARγ pathway in spontaneously hypertensive rats by Lixin Na; Xia Chu; Shuo Jiang; Chunjuan Li; Gang Li; Ying He; Yuanxiu Liu; Ying Li; Changhao Sun (1245-1253).
Vinegar has been reported to lower blood pressure, but its mechanism is unclear. This study explored whether vinegar plays antihypertensive effect by activating AMP-activated protein kinase (AMPK) pathway.Male spontaneously hypertensive rats (SHRs) were assigned to vinegar, acetic acid, nifedipine, nifedipine + vinegar, or distilled water by oral gavage for 8 weeks. Blood and aortas were analyzed for biochemical indices and protein expression levels. Sv40-transformed aortic rat endothelia cell line (SVAREC) cells were treated with acetate at different doses for 24 h; protein expression levels were assessed.Vinegar and acetic acid decreased blood pressure in SHRs on weeks 6 and 8, and nifedipine + vinegar had a better effect on blood pressure control than vinegar or nifedipine alone. Vinegar and acetic acid could decrease serum renin and angiotensin-converting enzyme (ACE) activities, angiotensin II and aldosterone concentrations in SHRs. Vinegar and acetic acid also increased AMP/ATP ratios and expression levels of pAMPK, PPARγ coactivator-1α (PGC-1α), and PPARγ while inhibited angiotensin II type 1 receptor (AT1R) expression in SHRs. The changes in these protein expressions were also found in SVAREC cells treated with 200 or 400 μmol/L acetate. In the presence of AMPK inhibitor or PGC-1α small interfering RNA, the effects of acetate on their downstream protein expression in SVAREC cells were abolished, respectively.Vinegar activates AMPK by increasing AMP/ATP ratios, thereby increases PGC-1α and PPARγ expressions, and inhibits AT1R expression in SHRs. Acetic acid is responsible for the antihypertensive effects of vinegar. There is a joint effect between vinegar and nifedipine in blood pressure control.
Keywords: AMPK; Angiotensin II type 1 receptor; Blood pressure; Vinegar

Mid-regional-pro-adrenomedullin plasma levels are increased in obese adolescents by Silvia Del Ry; Manuela Cabiati; Vanessa Bianchi; Laura Caponi; Pietro Di Cecco; Benedetta Marchi; Emioli Randazzo; Chiara Caselli; Tommaso Prescimone; Aldo Clerico; Giovanni Federico (1255-1260).
Recently, adrenomedullin (ADM) was defined as a new member of the adipokine family. ADM secreted by adipocytes, through its vasodilator and antioxidant actions, might be protective against metabolic syndrome-associated cardiovascular complications. The aim of the study was to assess plasma mid-regional (MR)-proADM levels in obese adolescents compared to normal-weight subjects and its relation with BMI, body composition and metabolic indices.Plasma MR-proADM was measured in 32 healthy adolescents [BMI z-score (mean ± SEM) = 0.6 ± 0.09 and 0.8 ± 0.07 in females and males, respectively] and in 51 age-matched obese adolescents [BMI z-score (mean ± SEM) = 2.8 ± 0.12 and 2.9 ± 0.08 in female and males, respectively] by a time-resolved amplified cryptate emission technology assay.Plasma MR-proADM levels resulted significantly higher in obese than in normal-weight adolescents (MR-proADM: 0.33 ± 0.1 vs 0.40 ± 0.1 nmol/L, p < 0.0001). Using univariate analysis, we observed that MR-proADM correlated significantly with BMI z-score (p < 0.0001), fat mass (p < 0.0001), circulating insulin (p < 0.004), HOMA-IR (p < 0.005), total cholesterol (p < 0.03) and LDL-cholesterol (p < 0.05). Including MR-proADM as response variable and its significant correlates into a multiple regression analysis, we observed that fat mass (p = 0.014) and BMI z-score (p = 0.036) were independent determinants of circulating MR-proADM.Our study shows for the first time that obese adolescents have higher circulating levels of MR-proADM compared with normal-weight, appropriate controls suggesting its important involvement in obese patients.
Keywords: MR-proADM; Obesity; Adolescents; Adipokine

Green tea polyphenol extract in vivo attenuates inflammatory features of neutrophils from obese rats by K. F. F. S. Albuquerque; M. P. Marinovic; A. C. Morandi; A. P. Bolin; R. Otton (1261-1274).
Our study aimed to evaluate whether obesity induced by cafeteria diet changes the neutrophil effector/inflammatory function and whether treatment with green tea extract (GT) can improve neutrophil function.Male Wistar rats were treated with GT by gavage (12 weeks/5 days/week; 500 mg/kg of body weight), and obesity was induced by cafeteria diet (8 weeks). Neutrophils were obtained from the peritoneal cavity (injection of oyster glycogen). The following analyses were performed: phagocytic capacity, chemotaxis, myeloperoxidase activity (MPO), hypochlorous acid (HOCl), superoxide anion (O 2 ·− ), hydrogen peroxide (H2O2), IL-1β, IL-6 and TNFα, mRNA levels of inflammatory genes, calcium mobilisation, activities of antioxidant enzymes, hexokinase and G6PDH. Neutrophils from obese rats showed a significant decrease in migration capacity, H2O2 and HOCl production, MPO activity and O 2 ·− production. Phagocytosis and CD11b mRNA levels were increased, while inflammatory cytokines release remained unmodified. mRNA levels of TLR4 and IκK were enhanced. Treatment of obese rats with GT increased neutrophil migration, MPO activity, H2O2, HOCl and O 2 ·− production, whereas TNF-α and IL-6 were decreased (versus obese). Similar reductions in TLR4, IκK and CD11b mRNA were observed. Catalase and hexokinase were increased by obesity, while SOD and G6PDH were decreased. Treatment with GT reduced catalase and increased the GSH/GSSG ratio.In response to a cafeteria diet, we found a decreased chemotaxis, H2O2 release, MPO activity and HOCl production. We also showed a significant immunomodulatory effect of GT on the obese condition recovering some of these factors such H2O2 and HOCl production, also reducing the levels of inflammatory cytokines.
Keywords: Obesity; Immune system; Polyphenols; Reactive oxygen species; Flavonoids; Catechins

Preliminary iodine concentration (UIC) measurements in spot urines of the representative German adult study DEGS indicated a severe worsening of iodine status compared to previous results in German children (KiGGS study). Therefore, we aimed to evaluate adult iodine status in detail and to investigate the impact of hydration status on UIC. UIC and creatinine concentrations were measured in 6978 spot urines from the German nationwide DEGS study (2008–2011). Twenty-four-hour iodine excretions (24-h UIE) were estimated by relating iodine/creatinine ratios to age- and sex-specific 24-h creatinine reference values. Urine osmolality was measured in two subsamples of spot urines (n = 100 each) to determine the impact of hydration status on UIC.In DEGS, median UIC was 69 µg/L in men and 54 µg/L in women, lying clearly below the WHO cutoff for iodine sufficiency (100 µg/L). Estimated median 24-h UIE was 113 µg/day, accompanied by 32 % of DEGS adults, lying below the estimated average requirement (EAR) for iodine. Comparative analysis with the KiGGS data (>14,000 spot urines of children; median UIC 117 µg/L) revealed a comparable percentage
Keywords: Iodine; DEGS study; Creatinine; Spot urine; 24-h Iodine excretion

Amelioration of oxidative and inflammatory status in hearts of cholesterol-fed rats supplemented with oils or oil-products with extra virgin olive oil components by Ageliki I. Katsarou; Andriana C. Kaliora; Antonia Chiou; Nick Kalogeropoulos; Apostolos Papalois; George Agrogiannis; Nikolaos K. Andrikopoulos (1283-1296).
The contribution of extra virgin olive oil (EVOO) macro- and micro-constituents in heart oxidative and inflammatory status in a hypercholesterolemic rat model was evaluated. Fatty acid profile as well as α-tocopherol, sterol, and squalene content was identified directly in rat hearts to distinguish the effect of individual components or to enlighten the potential synergisms.Oils and oil-products with discernible lipid and polar phenolic content were used. Wistar rats were fed a high-cholesterol diet solely, or supplemented with one of the following oils, i.e., EVOO, sunflower oil (SO), and high-oleic sunflower oil (HOSO) or oil-products, i.e., phenolics-deprived EVOO [EVOO(−)], SO enriched with the EVOO phenolics [SO(+)], and HOSO enriched with the EVOO phenolics [HOSO(+)]. Dietary treatment lasted 9 weeks; at the end of the intervention blood and heart samples were collected.High-cholesterol-diet-induced dyslipidemia was shown by increase in serum total cholesterol, low-density lipoprotein cholesterol, and triacylglycerols. Dyslipidemia resulted in increased malondialdehyde (MDA) and tumor necrosis factor-α (TNF-α) levels, while glutathione and interleukin 6 levels remained unaffected in all intervention groups. Augmentation observed in MDA and TNF-α was attenuated in EVOO, SO(+), and HOSO(+) groups. Heart squalene and cholesterol content remained unaffected among all groups studied. Heart α-tocopherol was determined by oil α-tocopherol content. Variations were observed for heart β-sitosterol, while heterogeneity was reported with respect to heart fatty acid profile in all intervention groups.Overall, we suggest that the EVOO-polar phenolic compounds decreased MDA and TNF-α in hearts of cholesterol-fed rats.
Keywords: Extra virgin olive oil; Hypercholesterolemia; Inflammatory markers; Oxidation; Polar phenols; Rat heart

Mediatory effect of circulating vaspin on resting metabolic rate in obese individuals by Sajjad Moradi; Khadijeh Mirzaei; Ahmed Abdulahi Abdurahman; Seyed Ali Keshavarz; Arash Hossein-nezhad (1297-1305).
Vaspin is a recently identified adipokine related to obesity and insulin sensitivity. The precise mechanism of vaspin in the body is not well known, and its function in resting metabolic rate (RMR) is even less understood. Therefore, the main aim of this study was to investigate the effect of circulating vaspin on RMR in obese people.A total of 222 obese participants were included in the current comparative cross-sectional study. Body composition was measured using body composition analyzer. RMR was measured by means of indirect calorimetry. For the measurement of vaspin serum concentrations, an enzyme-linked immunosorbent assay was used. Dietary intake was assessed using 3-day 24-h dietary recall.Between low and high circulating vaspin groups, there was significant difference for sex (P = 0.03), fat percent (P = 0.008), RMR per weight (P < 0.001), and RMR per fat free mass (FFM) (P = 0.007). However, there was no statistical difference between the groups in dietary intake after adjustment for energy intake (P > 0.05). Furthermore, individuals with higher level of RMR had higher vaspin concentration. Weight, visceral fat, FFM, and fat mass had significant effect on increasing RMR (P < 0.05) but after adding vaspin as a covariate in the general linear model; visceral fat (P = 0.078) and fat mass (P = 0.339) missed their effectiveness.Circulating vaspin level is higher in women than in men in obese individuals. Moreover, it was found that vaspin had mediator effect between visceral fat and fat mass associations with RMR in obese participants.
Keywords: Vaspin; Resting metabolic rate; Obese; Adipose tissue

This study was aimed to investigate the effects of a maternal low-protein diet on transcriptional regulation of the myostatin (MSTN) gene in skeletal muscle of weaning piglets.Sows were fed either a standard-protein (SP, 15 and 18 % crude protein) or a low-protein (LP, 50 % protein level of SP) diet throughout pregnancy and lactation. Longissimus dorsi muscle was sampled from male piglets at 28 days of age. The mRNA was determined by RT-PCR, and protein was measured by Western blot. Chromatin immunoprecipitation assay was used to determine the binding of transcription factors and histone H3 modifications on the MSTN gene promoter.The maternal LP diet significantly decreased body weight and average daily gain (P < 0.05), which was associated with significantly lower plasma concentration of urea nitrogen and total protein (P < 0.05), as well as decreased muscle RNA content (P < 0.05). MSTN mRNA (P < 0.05) was significantly increased, together with enhanced (P < 0.05) mRNA and protein expression of forkhead box class O family member protein 3 (FoxO3), and a tendency of an increase (P = 0.10) in glucocorticoid receptor (GR) mRNA in the muscle of LP piglets. Furthermore, the binding of both FoxO3 and GR to the MSTN gene promoter was significantly higher (P < 0.05) in muscle of LP piglets, together with significantly enriched (P < 0.05) gene activation markers, H3K9Ac and H3K4me3.These results indicate that MSTN mediates maternal LP diet-induced growth retardation, through epigenetic regulation involving FoxO3 and GR binding to its promoter.
Keywords: Histone modification; Low-protein diet; Myostatin; Skeletal muscle; Weaning piglets