European Journal of Nutrition (v.53, #1)

Diet, cognition, and Alzheimer’s disease: food for thought by Ane Otaegui-Arrazola; Pilar Amiano; Ana Elbusto; Elena Urdaneta; Pablo Martínez-Lage (1-23).
The prevention of Alzheimer’s disease (AD) has become a real challenge due to its rising prevalence and the lack of an effective cure. Diet and nutrients have gained significant interest as potentially modifiable protective factors.The aim of this review is to provide an updated summary of evidence related to the effect of diet and nutritional factors on the risk of AD and cognitive aging, and discuss the potential mechanisms and confounding factors involved.A search was conducted in Medline and Web of Knowledge for epidemiological and clinical studies in the international literature from January 2000 to February 2013 using combinations of the following keywords: “Alzheimer’s disease”, “mild cognitive impairment”, “cognitive function”, “dietary factors”, “omega-3”, “antioxidants”, “B vitamins”, “dietary patterns”, and “Mediterranean diet”.Data from observational studies point to a protective role for certain nutrients, such as omega-3 fatty acids, antioxidants or B vitamins, and dietary patterns (Mediterranean diet). However, data from randomized controlled trials do not show a consistent effect. Whether confounding factors such as age, disease stage, other dietary components, cooking processes, and other methodological issues explain the divergent results remains to be established. Moreover, if certain nutrients protect against dementia, it is as yet unknown whether they may have a general effect on brain vascular health or directly interfere with the etiopathogenesis of AD.
Keywords: Omega-3 fatty acids; Antioxidants; B vitamins; Mediterranean diet; Alzheimer’s disease

Exploring the benefits and challenges of establishing a DRI-like process for bioactives by Joanne R. Lupton; Stephanie A. Atkinson; Namsoo Chang; Cesar G. Fraga; Joseph Levy; Mark Messina; David P. Richardson; Ben van Ommen; Yuexin Yang; James C. Griffiths; John Hathcock (1-9).
Bioactives can be defined as: “Constituents in foods or dietary supplements, other than those needed to meet basic human nutritional needs, which are responsible for changes in health status” (Office of Disease Prevention and Health Promotion, Office of Public Health and Science, Department of Health and Human Services in Fed Reg 69:55821–55822, 2004). Although traditional nutrients, such as vitamins, minerals, protein, essential fatty acids and essential amino acids, have dietary reference intake (DRI) values, there is no such evaluative process for bioactives. For certain classes of bioactives, substantial scientific evidence exists to validate a relationship between their intake and enhanced health conditions or reduced risk of disease. In addition, the study of bioactives and their relationship to disease risk is a growing area of research supported by government, academic institutions, and food and supplement manufacturers. Importantly, consumers are purchasing foods containing bioactives, yet there is no evaluative process in place to let the public know how strong the science is behind the benefits or the quantitative amounts needed to achieve these beneficial health effects. This conference, Bioactives: Qualitative Nutrient Reference Values for Life-stage Groups?, explored why it is important to have a DRI-like process for bioactives and challenges for establishing such a process.
Keywords: Bioactives ; Dietary reference intakes; Non-essential nutrients; Adequate intake

Coffee and caffeine have been linked to type 2 diabetes mellitus (T2DM). A dose–response meta-analysis of prospective studies was conducted to assess the association between coffee and caffeine intake and T2DM incidence.Pertinent studies were identified by a search of PubMed and EMBASE. The fixed- or random-effect pooled measure was selected based on between-study heterogeneity. Dose–response relationship was assessed by restricted cubic spline.Compared with the lowest level, the pooled relative risk (95 % CI) of T2DM was 0.71 (0.67–0.76) for the highest level of coffee intake (26 articles involving 50,595 T2DM cases and 1,096,647 participants), 0.79 (0.69–0.91) for the highest level of decaffeinated coffee intake (10 articles involving 29,165 T2DM cases and 491,485 participants) and 0.70 (0.65–0.75) for the highest level of caffeine intake (6 articles involving 9,302 T2DM cases and 321,960 participants). The association of coffee, decaffeinated coffee and caffeine intake with T2DM incidence was stronger for women than that for men. A stronger association of coffee intake with T2DM incidence was found for non-smokers and subjects with body mass index <25 kg/m2. Dose–response analysis suggested that incidence of T2DM decreased by 12 % [0.88 (0.86–0.90)] for every 2 cups/day increment in coffee intake, 11 % [0.89 (0.82–0.98)] for every 2 cups/day increment in decaffeinated coffee intake and 14 % [0.86 (0.82–0.91)] for every 200 mg/day increment in caffeine intake.Coffee and caffeine intake might significantly reduce the incidence of T2DM.
Keywords: Coffee; Caffeine; Type 2 diabetes mellitus; Dose–response meta-analysis

A high intake of dietary fiber influences C-reactive protein and fibrinogen, but not glucose and lipid metabolism, in mildly hypercholesterolemic subjects by Anna Johansson-Persson; Matilda Ulmius; Lieselotte Cloetens; Toni Karhu; Karl-Heinz Herzig; Gunilla Önning (39-48).
The aim of the study was to investigate how a diet high in dietary fiber, with several fiber sources included, modulates glucose and lipid metabolism and the inflammatory response in humans.Subjects (n = 25) aged 58.6 (1.1) years (mean and SD) with a BMI of 26.6 (0.5) kg/m2 and a total cholesterol (TC) of 5.8 (0.1) mmol/L (mean and SEM) were given a high fiber (HF) and low fiber (LF) diet, in a randomized controlled 5-week crossover intervention, separated by a 3-week washout. The HF diet consisted of oat bran, rye bran, and sugar beet fiber incorporated into test food products; one bread roll, one ready meal, and two beverages consumed daily. Equivalent food products, without added fibers, were provided in the LF diet.Total dietary fiber intake was 48.0 g and 30.2 g per day for the HF and LF diet, respectively. Significant reduction in C-reactive protein (CRP) was observed between the diets (P = 0.017) and a significant reduction in fibrinogen within the HF diet (P = 0.044). There were no significant effects in other measured circulating cytokines or in glucose, insulin, and lipid levels.Our study suggests that a 5-week high dietary fiber intake of oat bran, rye bran, and sugar beet fiber might reduce the low-grade inflammatory response measured as CRP which could, together with reduced fibrinogen, help to prevent the risk of cardiovascular disease.
Keywords: Inflammatory markers; Dietary fiber; Oat; Rye; Sugar beet

Eating carbohydrate mostly at lunch and protein mostly at dinner within a covert hypocaloric diet influences morning glucose homeostasis in overweight/obese men by Raquel Duarte Moreira Alves; Fernanda Cristina Esteves de Oliveira; Helen Hermana Miranda Hermsdorff; Itziar Abete; María Ángeles Zulet; José Alfredo Martínez; Josefina Bressan (49-60).
To evaluate the effects of two dietary patterns in which carbohydrates and proteins were eaten mostly at lunch or dinner on body weight and composition, energy metabolism, and biochemical markers in overweight/obese men.Fifty-eight men (30.0 ± 7.4 years; 30.8 ± 2.4 kg/m2) followed a covert hypocaloric balanced diet (−10 % of daily energy requirements) during 8 weeks. Subjects were randomly assigned to three groups: control diet (CT); diurnal carbohydrate/nocturnal protein (DCNP); and nocturnal carbohydrate/diurnal protein (NCDP). Main analyzed outcomes were weight loss, body composition, diet-induced thermogenesis (DIT), and glucose/lipid profile.In all groups, a significant decrease in body weight, BMI, and fat mass (kg and %) was verified, without differences between groups. Interestingly, within group analyses showed that the fat-free mass (kg) significantly decreased in NCDP and in CT after 8-week intervention, but not in DCNP. A detrimental increase in fasting glucose, insulin, and homeostasis model assessment of insulin resistance (HOMAIR) was verified only in DCNP, while NCDP and CT groups presented a non-significant reduction. Moreover, significant differences between DCNP and the other groups were detected for fasting insulin and HOMAIR. After the adjustments, NCDP presented a significantly higher DIT and energy expenditure after lunch, compared with DCNP, but after dinner, there were no differences among groups.Eating carbohydrates mostly at dinner and protein mostly at lunch within a hypocaloric balanced diet had similar effect on body composition and biochemical markers, but higher effect on DIT compared with control diet. Moreover, eating carbohydrates mostly at lunch and protein mostly at dinner had a deleterious impact on glucose homeostasis.
Keywords: Obesity; Weight management; Body composition; Macronutrients; Energy metabolism; Glucose homeostasis

Few studies have focused on the metabolic changes induced by creatine supplementation. This study investigated the effects of creatine supplementation on plasma and urinary metabolite changes of athletes after endurance and sprint running.Twelve male athletes (20.3 ± 1.4 y) performed two identical (65–70 % maximum heart rate reserved) 60 min running exercises (endurance trial) before and after creatine supplementation (12 g creatine monohydrate/day for 15 days), followed by a 5-day washout period. Subsequently, they performed two identical 100 m sprint running exercises (power trial) before and after 15 days of creatine supplementation in accordance with the supplementary protocol of the endurance trial. Body composition measurements were performed during the entire study. Plasma samples were examined for the concentrations of glucose, lactate, branched-chain amino acids (BCAAs), free-tryptophan (f-TRP), glutamine, alanine, hypoxanthine, and uric acid. Urinary samples were examined for the concentrations of hydroxyproline, 3-methylhistidine, urea nitrogen, and creatinine.Creatine supplementation significantly increased body weights of the athletes of endurance trial. Plasma lactate concentration and ratio of f-TRP/BCAAs after recovery from endurance running were significantly decreased with creatine supplementation. Plasma purine metabolites (the sum of hypoxanthine and uric acid), glutamine, urinary 3-methylhistidine, and urea nitrogen concentrations tended to decrease before running in trials with creatine supplements. After running, urinary hydroxyproline concentration significantly increased in the power trial with creatine supplements.The findings suggest that creatine supplementation tended to decrease muscle glycogen and protein degradation, especially after endurance exercise. However, creatine supplementation might induce collagen proteolysis in athletes after sprint running.
Keywords: Purine metabolites; Glutamine; Alanine; Hydroxyproline; 3-Methylhistidine; Urinary urea nitrogen

Ilex paraguariensis (yerba mate) improves endocrine and metabolic disorders in obese rats primed by early weaning by Natália da S. Lima; Juliana G. Franco; Nayara Peixoto-Silva; Lígia A. Maia; Andrea Kaezer; Israel Felzenszwalb; Elaine de Oliveira; Egberto G. de Moura; Patricia Cristina Lisboa (73-82).
We showed that early weaned rats developed obesity, hyperleptinemia, leptin and insulin resistance at adulthood. Here, we studied the potential beneficial effects of Ilex paraguariensis aqueous solution upon body composition, glycemia, lipid and hormonal profiles, leptin signaling and NPY content.To induce early weaning, lactating rats’ teats were blocked with a bandage to interrupt lactation during the last 3 days (EW group), while control offspring had free access to milk throughout lactation (C group). In postnatal day (PN) 150, EW offspring were subdivided into: EW and EW+ mate groups treated, respectively, with water or yerba mate aqueous solution (1 g/kg BW/day, gavage) during 30 days. C offspring received water for gavage. In PN180, offspring were killed.EW+ mate group presented lower body weight (−10 %), adipose mass (retroperitoneal:−40 % and epididymal:−44 %), total body fat (−43 %), subcutaneous fat (−46 %), visceral adipocyte area (−21 %), triglyceridemia (−31 %) and hypothalamic NPY content (−37 %) compared to EW group. However, hyperglycemia and lower HDL-c levels observed in EW group were not reverted with mate treatment. Although the hyperleptinemia, lower hypothalamic JAK2 and pSTAT3 content of EW group were not corrected by mate treatment, the hyperphagia and higher hypothalamic SOCS-3 content were normalized in EW+ mate group, indicating that the central leptin resistance could be restored.Thus, the therapy with yerba mate solution was capable to reverse abdominal obesity, leptin resistance and hypertriglyceridemia, suggesting an important role of this bioactive component in the management of obesity in this programming model.
Keywords: Obesity; Programming; Fat mass; Leptin; Ilex paraguariensis

Effects of improved fat content of frankfurters and pâtés on lipid and lipoprotein profile of volunteers at increased cardiovascular risk: a placebo-controlled study by Gonzalo Delgado-Pando; Paloma Celada; Francisco J. Sánchez-Muniz; Francisco Jiménez-Colmenero; Begoña Olmedilla-Alonso (83-93).
To study the effect of modified frankfurters and pâtés: (a) reduced-fat products (RF) (15.3 and 15.2 % fat, respectively); (b) n-3-enriched reduced-fat products (n-3 RF) (15.1 and 15.5 % fat, respectively); and (c) normal-fat products (NF) (18 and 30.8 % fat, respectively) on lipids, lipoproteins, atherogenic ratios, oxidized LDL, and blood pressure of volunteers at high CVD risk.Twenty-two volunteers were enrolled in a sequential study of 3 consecutive 4-week periods separated by 4-week washout periods.LDL-cholesterol (p < 0.01), oxidized LDL, LDL-cholesterol/HDL-cholesterol (both p < 0.05) were significantly affected by the overall intervention. Compared to baseline, LDL-cholesterol decreased significantly (p = 0.012) during the RF period; the LDL-cholesterol/HDL-cholesterol ratio (p = 0.08) and the diastolic blood pressure (p = 0.06) also decreased, although non-significantly, after RF consumption. LDL-cholesterol (p = 0.040) and oxidized LDL (p = 0.016) increased significantly after NF product consumption; systolic blood pressure did not show significant variations after this period. No significant differences, in absolute or relative changes, were observed between RF and n-3 RF consumption for any parameter tested. However, LDL-cholesterol, oxidized LDL, and the LDL-cholesterol/HDL-cholesterol ratio were lower (12, 15 and 10 %, respectively) after n-3 RF versus NF product consumption. Oxidized LDL was approximately 15 % lower after RF versus NF product consumption.The regular consumption of RF meat products, enriched in n-3 fatty acids or not, positively affects the lipoprotein profile of volunteers, decreasing LDL-cholesterol and oxidized LDL levels and, thus, future risk of cardiovascular accident. On comparison with the effects of NF product intake, the responses to n-3 RF and RF products differ, and while n-3 RF intake induces a reduction in LDL-cholesterol, oxidized LDL-cholesterol, and LDL-cholesterol/HDL-cholesterol ratio, the intake of RF products modifies only the oxidized LDL level.
Keywords: Reduced-fat; Omega-3-enriched meat; Cholesterol; Lipids; Lipoprotein; Atherogenic ratio; Blood pressure; Frankfurters; Pâtés

Effectiveness of a tailor-made weight loss intervention in primary care by Bastiaan C. de Vos; Jos Runhaar; Sita M. A. Bierma-Zeinstra (95-104).
Weight loss interventions have been studied extensively, but methodological limitations negatively affecting applicability in everyday clinical practice are a very common problem in these studies. Despite the fact that obesity is treated mostly in a primary care setting, studies that investigate weight loss interventions in a primary care setting are scarce. Our objective was to assess the effectiveness of a tailor-made weight loss intervention in achieving a clinically significant weight loss in overweight (BMI ≥ 27 kg/m²) women aged 50–60 years in a primary care setting.As part of a randomized controlled trial on the effects of a tailor-made weight loss intervention and oral glucosamine sulphate on the incidence of osteoarthritis of the knee in 407 overweight women aged 50–60 years, we analysed the effectiveness of the weight loss intervention in achieving clinically relevant weight loss.At baseline, the mean body weight for all participants was 88.7 ± 13.2 kg, and the mean BMI was 32.4 ± 4.3 kg/m². The percentage of participants that lost ≥5 kg or 5 % of their baseline body weight was 14.8 versus 6.3 % (p = 0.012) at 6 months for the intervention group and the control group, respectively. At 12 months, this was 18.7 versus 14.9 % (p = 0.027). Mean weight gain at 6 months was −0.9 versus 0.9 kg (p < 0.001) for the intervention group and the control group, respectively. At 12 months, this was −0.6 versus 0.6 kg (p = 0.01). At 30 months of follow-up, no significant differences were found between both groups.This weight loss intervention, which, at short notice, is easily applicable in everyday clinical practice, is effective in achieving clinically significant weight loss in overweight women aged 50–60 over a 12-month period. Long-term weight loss maintenance, however, occurred only marginally. Magnitude of the effect is comparable to that achieved in many other more intensive weight loss interventions.
Keywords: Overweight; Weight reduction; Weight loss intervention

Fermented milk is considered one of the best sources for efficient consumption of probiotic strains by hosts to promote good health. The purpose of this study was to investigate the effects of orally administering LGG-fermented milk (LGG milk) on intestinal inflammation and injury and to study the mechanisms of LGG milk’s action.LGG milk and non-LGG-fermented milk (non-LGG milk) were administered through gavage to mice before and during dextran sodium sulfate (DSS)-induced intestinal injury and colitis. Inflammatory/injury score and colon length were assessed. Intestinal epithelial cells were treated with the soluble fraction of LGG milk to detect its effects on the epidermal growth factor receptor (EGFR) and its downstream target, Akt activation, cytokine-induced apoptosis, and hydrogen peroxide (H2O2)-induced disruption of tight junctions.LGG milk treatment significantly reduced DSS-induced colonic inflammation and injury, and colon shortening in mice, compared to that in non-LGG milk-treated and -untreated mice. The soluble fraction of LGG milk, but not non-LGG milk, stimulated the activation of EGFR and Akt in a concentration-dependent manner, suppressed cytokine-induced apoptosis, and attenuated H2O2-induced disruption of tight junction complex in the intestinal epithelial cells. These effects of LGG milk were blocked by the EGFR kinase inhibitor. LGG milk, but not non-LGG milk, contained two soluble proteins, p40 and p75, that have been reported to promote survival and growth of intestinal epithelial cells through the activation of EGFR. Depletion of p40 and p75 from LGG milk abolished the effects of LGG milk on prevention of cytokine-induced apoptosis and H2O2-induced disruption of tight junctions.These results suggest that LGG milk may regulate intestinal epithelial homeostasis and potentially prevent intestinal inflammatory diseases through activation of EGFR by LGG-derived proteins.
Keywords: Lactobacillus GG; Fermented milk; Colitis; Epidermal growth factor receptor; Apoptosis; Tight junction; Intestinal epithelial cells

Olive oil improves the intestinal absorption and bioavailability of lutein in lutein-deficient mice by Bhatiwada Nidhi; Bangera Sheshappa Mamatha; V. Baskaran (117-126).
To investigate the influence of olive (OO), groundnut (GNO), soybean (SBO), sunflower (SFO), rice bran (RBO), corn (CO), palm (PO) oil or mixed micelle (control) on absorption kinetics and bioavailability of lutein in lutein-deficient mice. Additional aim was to correlate the activity of intestinal triacylglycerol lipase with intestinal and plasma lutein levels.After induction of lutein deficiency, mice (n = 165) were divided into eight groups (OO, SFO, GNO, RBO, PO, CO, SBO and control; n = 20/group) and the remaining (n = 5) were used as baseline (0 h). Groups were further divided into four subgroups (n = 5/subgroup) and were intubated with lutein (200 μM) dispersed in different vegetable oils. Plasma and tissue (intestine, liver and eyes), lutein, triglycerides, intestinal triacylglycerol lipases and fatty acid profile of plasma and tissues were measured at different time intervals.The percentage area under the curve value for plasma lutein in OO and GNO was higher by 41.8 and 5.1 %, while it was lower in other groups (18.2–53.3 %), when compared to control. Similarly, the percentage area under the curve for eye lutein in OO and GNO groups was higher by 35.2 and 4.8 %, whereas in other groups it was lower (5.4–69 %) than in control. Results show that olive oil facilitates the lutein absorption more compared to other vegetable oils, which may be due to the difference in fatty acid composition and higher activity of intestinal triacylglycerol lipase.Dietary olive oil rich in oleic acid improves the bioavailability and accumulation of lutein in lutein-deficient mice by modifying the intestinal triacylglycerol lipase activity.
Keywords: Absorption kinetics; Bioavailability; Lutein deficiency; Triacylglycerol lipase; Vegetable oils

The effect of chromium on inflammatory markers, 1st and 2nd phase insulin secretion in type 2 diabetes by Yen-Lin Chen; Jiunn-Diann Lin; Te-Lin Hsia; Frank Chiahung Mao; Chun-Hsien Hsu; Dee Pei (127-133).
Impaired insulin sensitivity (SI) and β-cell function are the two main causes of type 2 diabetes (T2D) and are related to low-grade inflammation status. Trivalent chromium has shown to improve SI in our previous study. This might be due to the ability of decreasing interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) shown in animal studies. In the current study, we measured SI, β-cell function, and plasma levels of IL-6 and TNF-α after treatment of chromium chloride (GaCr) in T2D.Sixty-six patients were randomly assigned to the 20 g of GaCr milk powder studying group or the milk powder placebo group. Oral glucose tolerance test was performed before and after the treatment. The SI and the β-cell function were measured as well.The SI was significantly improved. At the same time, the static insulin responsivity index (Φs) was significantly higher after the treatment (p = 0.003). On the other hand, the dynamic insulin responsivity index (Φd) remained unchanged. Interestingly, a significant decrease in the IL-6 level after the treatment (p = 0.015) was noted. Although there was a trend of decreasing in TNF-α, it was not statistically significant. Finally, there was no significant correlation between the δ-IL-6, SI, and Φd after GaCr treatment.In conclusion, other than the improvement of SI, GaCr could also improve the second phase of insulin responsivity (Φs) and IL-6. However, δ-IL-6 was correlated with neither δ-SI nor δ-Φs which indicated that the improvement of SI and Φs might involve mechanisms other than lower inflammatory effect.
Keywords: Chromium; Insulin sensitivity; Insulin secretion; Interleukin-6; Tumor necrosis factor-α

Inflammation is a hallmark of many diseases, such as atherosclerosis, autoimmune diseases, obesity, and cancer. Isoflavone-free soy protein diet (SPI) has been shown to reduce atherosclerotic lesions in a hyperlipidemic mouse model compared to casein (CAS)-fed mice, despite unchanged serum lipid levels. However, possible mechanisms contributing to the athero-protective effect of soy protein remain unknown. Therefore, we investigated whether and how SPI diet inhibits inflammatory responses associated with atherosclerosis.Apolipoprotein E knockout (apoE−/−) mice (5-week) were fed CAS or SPI diet for 1 or 5 week to determine LPS- and hyperlipidemia-induced acute and chronic inflammatory responses, respectively. Expression of NF-κB-dependent inflammation mediators such as VCAM-1, TNF-α, and MCP-1 were determined in aorta and liver. NF-κB, MAP kinase, and AKT activation was determined to address mechanisms contributing to the anti-inflammatory properties of soy protein/peptides.Isoflavone-free soy protein diet significantly reduced LPS-induced VCAM-1 mRNA and protein expression in aorta compared to CAS-fed mice. Reduced VCAM-1 expression in SPI-fed mice also paralleled attenuated monocyte adhesion to vascular endothelium, a critical and primary processes during inflammation. Notably, VCAM-1 mRNA and protein expression in lesion-prone aortic arch was significantly reduced in apoE−/− mice fed SPI for 5 weeks compared with CAS-fed mice. Moreover, dietary SPI potently inhibited LPS-induced NF-κB activation and the subsequent upregulation of pro-inflammatory cytokines, including TNF-α, IL-6, IL-1β, and MCP-1. Interestingly, SPI inhibited NF-κB-dependent inflammatory responses by targeting I-κB phosphorylation and AKT activation with no effect on MAP kinase pathway. Of the five putative soy peptides, four of the soy peptides inhibited LPS-induced VCAM-1, IL-6, IL-8, and MCP-1 protein expression in human vascular endothelial cells in vitro.Collectively, our findings suggest that anti-inflammatory properties of component(s) of soy protein/peptides may be a possible mechanism for the prevention of chronic inflammatory diseases such as atherosclerosis.
Keywords: Soy peptides; Inflammation; VCAM-1; Hyperlipidemia; Atherosclerosis

Dietary cocoa ameliorates obesity-related inflammation in high fat-fed mice by Yeyi Gu; Shan Yu; Joshua D. Lambert (149-158).
To investigate the effect of cocoa powder supplementation on obesity-related inflammation in high fat (HF)-fed obese mice.Male C57BL/6J (n = 126) were fed with either low-fat (LF, 10 % kcal from fat) or HF (60 % kcal from fat) diet for 18 weeks. After 8 weeks, mice from HF group were randomized to HF diet or HF diet supplemented with 8 % cocoa powder (HF–HFC group) for 10 weeks. Blood and tissue samples were collected for biochemical analyses.Cocoa powder supplementation significantly reduced the rate of body weight gain (15.8 %) and increased fecal lipid content (55.2 %) compared to HF-fed control mice. Further, cocoa supplementation attenuated insulin resistance, as indicated by improved HOMA-IR, and reduced the severity of obesity-related fatty liver disease (decreased plasma alanine aminotransferase and liver triglyceride) compared to HF group. Cocoa supplementation also significantly decreased plasma levels of the pro-inflammatory mediators interleukin-6 (IL-6, 30.4 %), monocyte chemoattractant protein-1 (MCP-1, 25.2 %), and increased adiponectin (33.7 %) compared to HF-fed mice. Expression of pro-inflammatory genes (Il6, Il12b, Nos2, and Emr1) in the stromal vascular fraction (SVF) of the epididymal white adipose tissue (WAT) was significantly reduced (37–56 %) in the cocoa-supplemented mice.Dietary supplementation with cocoa ameliorates obesity-related inflammation, insulin resistance, and fatty liver disease in HF-fed obese mice, principally through the down-regulation of pro-inflammatory gene expression in WAT. These effects appear to be mediated in part by a modulation of dietary fat absorption and inhibition of macrophage infiltration in WAT.
Keywords: Theobroma cacao ; Cocoa; Polyphenols; Inflammation; Obesity

Absorption and isomerization of caffeoylquinic acids from different foods using ileostomist volunteers by T. Erk; M. Renouf; G. Williamson; R. Melcher; H. Steiling; E. Richling (159-166).
Polyphenols are thought to play important roles in human nutrition and health but these health effects are dependent on their bioavailability. This study is one of a series with the aim of determining possible effects of food matrices on caffeoylquinic acid (CQA) bioavailability using ileostomy volunteers.After a CQA-free diet, ileostomists consumed coffee (746 μmol total CQA), and CQAs in excreted ileal fluid were subsequently identified and quantified with HPLC–diode array detection and HPLC-ESI-MS/MS. In our previous studies, other food sources such as cloudy apple juice (CAJ) (358 μmol CQA) and apple smoothie (AS) (335 μmol CQA) were investigated with the same model.Interesterification of CQA from both apple matrices was observed during gastrointestinal passage, whereas CQA consumed in coffee was not influenced by interesterification reactions. In total, 74.3, 22.4, and 23.8 % of the CQA from CAJ, AS, and coffee, respectively, were absorbed or degraded.Our results show that variations in food matrices and variations in phenolic composition have a major influence on intestinal bioavailability and interesterification of the investigated subclass of polyphenols, the CQAs.
Keywords: Caffeoylquinic acid; Apple juice; Apple smoothie; Coffee; Bioavailability; Ileostomy

The Czech Republic lacks body composition data for women. Therefore, the purpose of the study was to analyze body composition [body fat mass (BFM), fat-free mass (FFM), body fat percentage (%BFM) and visceral adipose tissue (VAT)] and to evaluate the changes that occur with aging in women aged 18–89 years. We also analyzed anthropometric characteristics of study participants and developed age-specific percentile curves for body composition parameters.A cross-sectional, non‐randomized study was conducted with a sample of 1,970 apparently healthy Czech women. Body composition was measured using a direct segmental multi-frequency bioelectrical impedance analysis (BSM-BIA).The mean BFM was 19.7 ± 8.9 kg, and BFM reached its peak in women over 70, at 27.6 ± 8.8 kg. There was a strong correlation between BFM and age (r = 0.61; r 2 = 0.37). Fifty percent of the women in the study had a BFM between 13.0 and 25.0 kg. The %BFM (r = 0.69; r 2 = 0.47) and VAT (r = 0.88; r 2 = 0.77) were also significantly associated with age. The reference range for %BFM was 22.0–35.6 % (25th–75th percentile). The mean FFM was 45.8 ± 5.5 kg, and FFM decreased with age (r = −0.27; r 2 = 0.07).The results presented in this study showed a statistically significant increase in BFM,  %BFM and VFA as age increased, and the values reached their peak in women over 70. Even when FFM decreased slightly with age, body weight increased because of the increase in BFM.
Keywords: Body fat mass; Fat-free mass; Visceral adipose tissue; Percentiles; InBody 720

Effect of ascorbic acid deficiency on catecholamine synthesis in adrenal glands of SMP30/GNL knockout mice by Akiko Amano; Makoto Tsunoda; Toshiro Aigaki; Naoki Maruyama; Akihito Ishigami (177-185).
The effect of an AA deficiency on catecholamine biosynthesis in adult mice in vivo is unknown. Therefore, we quantified catecholamine and the expression of catecholamine synthetic enzymes in the adrenal glands of senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice placed in an AA-deficient state.At 30 days of age, mice were divided into the following 4 groups: AA (−) SMP30/GNL KO, AA (+) SMP30/GNL KO, AA (−) wild type (WT), and AA (+) WT. The AA (+) groups were given water containing 1.5 g/L AA, whereas the AA (−) groups received water without AA until the experiment ended. In addition, all mice were fed an AA-depleted diet. Catecholamine levels were measured by a liquid chromatographic method. Tyrosine hydroxylase, dopa decarboxylase, dopamine β-hydroxylase, and phenylethanolamine N-methyltransferase mRNA expression levels were measured with the quantitative real-time polymerase chain reaction (qPCR). Tyrosine hydroxylase and dopamine β-hydroxylase protein levels were quantified by Western blot analysis.In the adrenals of AA (−) SMP30/GNL KO mice, noradrenaline and adrenaline levels decreased significantly compared to other three groups of mice, although there were no significant differences in dopamine β-hydroxylase or phenylethanolamine N-methyltransferase mRNA content. Moreover, there was no significant difference in their dopamine β-hydroxylase protein levels. On the other hand, AA depletion did not affect dopamine levels in adrenal glands of mice.An AA deficiency decreases the noradrenaline and adrenaline levels in adrenal glands of mice in vivo.
Keywords: Ascorbic acid; Catecholamines; Noradrenaline; Adrenaline; Dopamine; Adrenal glands

Epigallocatechin gallate attenuates fibrosis, oxidative stress, and inflammation in non-alcoholic fatty liver disease rat model through TGF/SMAD, PI3 K/Akt/FoxO1, and NF-kappa B pathways by Jia Xiao; Chi Tat Ho; Emily C. Liong; Amin A. Nanji; Tung Ming Leung; Thomas Yue Huen Lau; Man Lung Fung; George L. Tipoe (187-199).
To investigate the protective mechanisms of an 85 % pure extract of (−) epigallocatechin gallate (EGCG) in the development of fibrosis, oxidative stress and inflammation in a recently developed dietary-induced animal model of non-alcoholic fatty liver disease (NAFLD).Female Sprague–Dawley rats were fed with either normal rat diet or high-fat diet for 8 weeks to develop NAFLD. For both treatments, rats were treated with or without EGCG (50 mg/kg, i.p. injection, 3 times per week). At the end, blood and liver tissue samples were obtained for histology, molecular, and biochemical analyses.Non-alcoholic fatty liver disease (NAFLD) rats showed significant amount of fatty infiltration, necrosis, fibrosis, and inflammation. This was accompanied by a significant expressional increase in markers for fibrosis, oxidative stress, and inflammation. TGF/SMAD, PI3 K/Akt/FoxO1, and NF-κB pathways were also activated. Treatment with EGCG improved hepatic histology (decreased number of fatty score, necrosis, and inflammatory foci), reduced liver injury (from ~0.5 to ~0.3 of ALT/AST ratio), attenuated hepatic changes including fibrosis (reduction in Sirius Red and synaptophysin-positive stain) with down-regulation in the expressions of key pathological oxidative (e.g. nitrotyrosine formation) and pro-inflammatory markers (e.g. iNOS, COX-2, and TNF-α). EGCG treatment also counteracted the activity of TGF/SMAD, PI3 K/Akt/FoxO1, and NF-κB pathways. Treatment with EGCG did not affect the healthy rats.Epigallocatechin gallate (EGCG) reduced the severity of liver injury in an experimental model of NAFLD associated with lower concentration of pro-fibrogenic, oxidative stress, and pro-inflammatory mediators partly through modulating the activities of TGF/SMAD, PI3 K/Akt/FoxO1, and NF-κB pathways. Therefore, green tea polyphenols and EGCG are useful supplements in the prevention of NAFLD.
Keywords: EGCG; NAFLD; Fibrosis; Oxidative stress; Inflammation

Different postprandial acute response in healthy subjects to three strawberry jams varying in carbohydrate and antioxidant content: a randomized, crossover trial by Idoia Ibero-Baraibar; Marta Cuervo; Santiago Navas-Carretero; Itziar Abete; M. Angeles Zulet; J. Alfredo Martinez (201-210).
Dietary food composition influences postprandial glucose homeostasis. Thus, the objective was to investigate the effects of an acute intake of three different types of strawberry jam, differing in carbohydrate and antioxidants content, on postprandial glucose metabolism, lipid profile, antioxidant status, and satiety.Sixteen healthy adults participated in a randomized, crossover, double-blind study with three arms, receiving 60 g of three different strawberry jams. Blood samples were collected at fasting and at 30, 60, 90, and 120 min after its intake. Blood analyses were performed with validated procedures and satiety was estimated with visual analogue scale (VAS).Blood glucose concentrations were maintained at normal values and without peaks within the 2 h after consumption of low-sugar jams. However, blood glucose and insulin were significantly higher at 30 and 60 min after high-sugar (HS) jam intake versus both low-sugar jams. Furthermore, HS jam produced more satisfaction at short time, but decreased as soon as blood glucose concentration began to decrease. Moreover, HS ingestion produced lower free fatty acid levels (p < 0.05) throughout the trial with respect both the low-sugar jams. However, no additional benefits on oxidative status (malondialdehyde, glutathione peroxidase, total antioxidant capacity, and uric acid), glucose, lipid, and satiety variables were observed due to the inclusion of an antioxidant to low-sugar jam.This study reinforces the idea that products without added sugars are appropriate for the management of glycemic alterations and provides further insight into the effect of natural antioxidants as a functional ingredient on oxidative status and related metabolic disturbances. Registered at www.clinicaltrials.gov as NCT01684332.
Keywords: Antioxidant; Glucose metabolism; Jam; Polyphenols; Postprandial; Strawberry

Dietary fat quality in regular fat diets has minor effects on biomarkers of inflammation in obese Zucker rats by Daniela Graf; Stephan W. Barth; Achim Bub; Judith Narr; Corinna E. Rüfer; Bernhard Watzl; Stephanie Seifert (211-219).
Adipose tissue-associated chronic inflammation is involved in the pathogenesis of obesity-related diseases. Dietary fatty acids are known to influence inflammatory processes. The aim of this study was to investigate, whether diets with regular fat contents but variable fat qualities affect adipose tissue-associated inflammation through the fatty acid composition of mesenteric adipose tissue (MAT).Obese Zucker rats were fed diets containing 7 % wt:wt rapeseed oil, corn oil, or lard for 10 weeks. Fatty acid composition and endocrine function regarding adipokines and cytokines of MAT, number of total CD3+ T cells, and cytokine secretion of mesenteric lymph node (MLN)-derived lymphocytes were determined. Local effects in MAT and MLN were compared to systemic effects assessed in serum and peripheral blood mononuclear cells.Fatty acid composition of MAT reflected dietary fatty acid intake, without affecting endocrine function. Feeding the lard diet for 10 weeks increased the serum adiponectin and TNF-α secretion of blood lymphocytes, whereas CD3+ T cells in blood were decreased. No effects were seen for the secretion of adipokines and cytokines from MAT, the amount of T cells in MLN, and cytokine secretion of MLN lymphocytes.In conclusion, feeding obese rats a diet with regular fat content but variable fat sources for 10 weeks, changed the fatty acid composition of MAT but not its secretory properties or MLN functions. Although the local immune system was not influenced, lard-feeding induced minor changes in systemic immune function.
Keywords: Inflammation; Fat quality; Obesity; Immune function

Probiotic Lactobacillus casei Zhang ameliorates high-fructose-induced impaired glucose tolerance in hyperinsulinemia rats by Yong Zhang; Lifeng Wang; Jiachao Zhang; Yunxu Li; Qiuwen He; He Li; Xiao Guo; Jianlin Guo; Heping Zhang (221-232).
To evaluate the preventive and therapeutic effects of Lactobacillus casei Zhang on impaired glucose tolerance (IGT) by using fructose-induced hyperinsulinemia rats.Rats were fed 25 % fructose solution for hyperinsulinemia with L. casei Zhang for prevention or therapy. Serum levels of insulin, glucagon-like peptide-2 (GLP-2), osteocalcin, malondialdehyde (MDA), total intestinal bile acids and hepatic glycogen contents were determined by assay kits. The major bacteria from feces and liver expression of adiponectin receptor 2 (AdipoR2), liver X receptor-α (LXR-α), peroxisome proliferator-activated receptor gamma (PPAR-γ) and vitamin K epoxide reductase complex subunit 1 mRNA were assessed by RT-PCR. Pancreas injury was evaluated by histological analysis. Lactobacillus casei Zhang significantly increased numbers of Lactobacillus and Bifidobacterium and decreased Clostridium in the intestine (p < 0.01). Meanwhile, liver glycogen contents were significantly decreased (p < 0.05). In preventive group, accompanied by significantly lower insulin and GLP-2 levels (p < 0.05), L. casei Zhang prevented rats from an increase in oral glucose tolerance area under curve (AUC) which was significant in hyperinsulinemia group (p < 0.05). In therapeutic group, L. casei Zhang administration possessed improved glucose tolerance (p < 0.05), which were associated with increased osteocalcin level (p < 0.01), improved intestinal bile acids secretion (p = 0.060), decreased serum MDA levels (p < 0.05) and upregulation of LXR-α, PPAR-γ and AdipoR2 gene expression, as well as an increase in Bacteroides fragilis (p < 0.05). Lactobacillus casei Zhang administration exert both preventive and ameliorative effect on oral glucose tolerance AUC in IGT rats but may be via different mechanisms. L. casei Zhang could prevent rats from increased AUC through GLP-2 lowering, while the ameliorative effect in high-fructose-fed post-adolescent rats may be via B. fragilis enriched vitamin K2-dependent osteocalcin mechanism in which AdipoR2, LXR-α and PPAR-γ signaling were involved.
Keywords: Hyperinsulinemia rat; Lactobacillus casei Zhang; Osteocalcin; Microflora; Gene expression

The inhibitory activities of the edible green alga Capsosiphon fulvescens on rat lens aldose reductase and advanced glycation end products formation by Md. Nurul Islam; Sung Hwa Choi; Hye Eun Moon; Jin Ju Park; Hyun Ah Jung; Mi Hee Woo; Hee Chul Woo; Jae Sue Choi (233-242).
Accumulating evidence suggests that inhibitors of aldose reductase (AR) may prevent hyperglycemia-induced long-term complications in diabetes mellitus. In the present study, we evaluated the AR inhibitory potential of ethanolic (EtOH) extracts from 22 seaweed species.AR inhibitory activities of the selected seaweed species were evaluated using the rat lens aldose reductase (RLAR) inhibitory assay.All extracts exhibited RLAR inhibitory activity, which ranged from 5.87 to 92.71 % at a concentration of 50 μg/mL. Since Capsosiphon fulvescens exhibited significant inhibitory potential and is a frequently used foodstuff, it was selected for a detailed investigation using RLAR and advanced glycation end products (AGE) formation inhibitory assays. Among the different solvent-soluble fractions, the CH2Cl2, EtOAc, and n-BuOH fractions showed promising RLAR and AGE formation inhibitory activities. Considering the AR inhibitory potential, CH2Cl2 and EtOAc fractions were selected for chromatographic separation and yielded 11 compounds in which capsofulvesin A, capsofulvesin B, and chalinasterol showed potential RLAR inhibitory activity with the respective IC50 values of 52.53, 101.92, and 345.27 μM. Kinetic studies revealed that capsofulvesin A and chalinasterol exhibited mixed type inhibition, while capsofulvesin B exhibited noncompetitive inhibition. To our knowledge, this is the first report of AR inhibitory activity of the glycolipids capsofulvesin A and capsofulvesin B.Our results clearly indicate the potential RLAR and AGE formation inhibitory activities of C. fulvescens as well as its isolated constituents, which could be further explored to develop therapeutic modalities for the treatment of diabetes and related complications.
Keywords: Capsosiphon fulvescens ; Rat lens aldose reductase; Advanced glycation end products; Diabetic complications; Green alga; Glycolipid

Short bouts of anaerobic exercise increase non-esterified fatty acids release in obesity by Alberto Salvadori; Paolo Fanari; Paolo Marzullo; Franco Codecasa; Ilaria Tovaglieri; Mauro Cornacchia; Amelia Brunani; Livio Luzi; Erminio Longhini (243-249).
It is demonstrated that aerobic exercise plays an important role in weight loss programs for obesity by increasing 24 h metabolic rate. While aerobic exercise can result in health and fitness benefits in obese subjects, also independently of weight loss, not completely clear are the effects of bouts of hard exercise on metabolic outcomes. The aim of this study was to test the hypothesis that short-term aerobic activity with anaerobic bouts might result in a greater improvement in the management of obesity than aerobic activity alone.We studied 16 obese subjects (eight men) during a progressive cycloergometric test up to exhaustion, before and after 4 weeks of two different training schedules (6 days/week). Insulin and glycaemia, non-esterified fatty acids (NEFA) and lactic acid were sampled. Group A (eight subjects, four men) performed an aerobic cycle workout; Group B (eight subjects, four men) performed a 25 min aerobic workout followed by 5 min of anaerobic workout. All the subjects maintained their individual eating habits.The post-training test showed a decrease in AUCs NEFA in Group A (p < 0.05) and an increase in Group B (p < 0.05), together with an increase in lactic acid in Group A and a decrease in Group B (p < 0.01). β-cell function (HOMA2-B) revealed a reduction only in Group A (p < 0.05). Group B achieved a greatest reduction in body fat mass than Group A (p < 0.05).Aerobic plus anaerobic training seem to produce a greater response in lipid metabolism and not significant modifications in glucose indexes; then, in training prescription for obesity, we might suggest at starting weight loss program aerobic with short bouts of anaerobic training to reduce fat mass and subsequently a prolonged aerobic training alone to ameliorate the metabolic profile.
Keywords: Obesity; Exercise; Anaerobic threshold; Non-esterified fatty acids

Sugar-sweetened beverage and diet soda consumption and the 7-year risk for type 2 diabetes mellitus in middle-aged Japanese men by M. Sakurai; K. Nakamura; K. Miura; T. Takamura; K. Yoshita; S. Y. Nagasawa; Y. Morikawa; M. Ishizaki; T. Kido; Y. Naruse; Y. Suwazono; S. Sasaki; H. Nakagawa (251-258).
This cohort study investigated the association between sugar-sweetened beverage (SSB) and diet soda consumption and the incidence of type 2 diabetes in Japanese men.The participants were 2,037 employees of a factory in Japan. We measured consumption of SSB and diet soda using a self-administered diet history questionnaire. The incidence of diabetes was determined in annual medical examinations over a 7-year period. Hazard ratios (HRs) with 95 % confidence intervals (CIs) for diabetes were estimated after adjusting for age, body mass index, family history, and dietary and other lifestyle factors.During the study, 170 participants developed diabetes. The crude incidence rates (/1,000 person-years) across participants who were rare/never SSB consumers, <1 serving/week, ≥1 serving/week and <1 serving/day, and ≥1 serving/day were 15.5, 12.7, 14.9, and 17.4, respectively. The multivariate-adjusted HR compared to rare/never SSB consumers was 1.35 (95 % CI 0.80–2.27) for participants who consumed ≥1 serving/day SSB. Diet soda consumption was significantly associated with the incident risk of diabetes (P for trend = 0.013), and multivariate-adjusted HRs compared to rare/never diet soda consumers were 1.05 (0.62–1.78) and 1.70 (1.13–2.55), respectively, for participants who consumed <1 serving/week and ≥1 serving/week.Consumption of diet soda was significantly associated with an increased risk for diabetes in Japanese men. Diet soda is not always effective at preventing type 2 diabetes even though it is a zero-calorie drink.
Keywords: Cohort study; Epidemiology; Incidence; Nutrition

Increased vitamin D-binding protein and decreased free 25(OH)D in obese women of reproductive age by Therese Karlsson; Amra Osmancevic; Nina Jansson; Lena Hulthén; Agneta Holmäng; Ingrid Larsson (259-267).
Obese subjects have lower circulating 25-hydroxyvitamin D (25(OH)D) than normal-weight subjects. Knowledge is scarce regarding differences in vitamin D-binding protein (DBP), free 25(OH)D, and intake of vitamin D between normal-weight and obese subjects. The purpose of this study was to examine intake and vitamin D status in obese compared with normal-weight women.Between September 2009 and October 2011, 43 obese and 43 normal-weight women, 22–45 years of age, mean BMI of 39.1 ± 4.6 and 21.6 ± 1.8 kg/m2, respectively, were recruited in the western Sweden region (latitude 57°N). Blood samples, data regarding diet, and sun exposure were collected.DBP concentrations were 320 ± 121 and 266 ± 104 μg/mL (P = 0.02) in obese and normal-weight women, respectively. Calculated free 25(OH)D was 13.3 ± 5.5 (obese) and 23.7 ± 10.7 (normal-weight) (P < 0.001). The obese women had a 20.1 nmol/L lower mean 25(HO)D concentration compared to normal-weight women (P < 0.001). 56 % of obese women and 12 % of normal-weight women had 25(OH)D concentrations ≤50 nmol/L. There was no statistically significant difference in total vitamin D intake between the groups. 39 % of the women had a total vitamin D intake <7.5 μg/day, the current national recommendation for vitamin D in Sweden.Obese women had higher DBP concentrations compared with normal-weight women and lower free 25(OH)D. The obese women were more likely to have 25(OH)D concentrations that could be considered suboptimal. Vitamin D intake was generally low in normal-weight and obese women of childbearing age.
Keywords: Vitamin D; Vitamin D intake; Vitamin D status; Vitamin D-binding protein; Obesity; 25(OH)D

The flavonoid luteolin induces nitric oxide production and arterial relaxation by Hongwei Si; Richard P. Wyeth; Dongmin Liu (269-275).
Luteolin, a flavone present in many foods and medicinal plants, may have beneficial effects on various human chronic diseases. In the present study, we investigated the hypothesis that luteolin can directly act on vascular endothelial cells (ECs), leading to nitric oxide (NO) production and subsequent vascular relaxation.Rat aortic rings were mounted in organ bath. Luteolin was added cumulatively, and vessel relaxation of rat aortic rings precontracted with phenylephrine (PE) or potassium was recorded. Endothelial nitric oxide synthase (eNOS) phosphorylation at Ser1177 and NO production from aortic rings and primary human aortic endothelial cells (HAECs) exposed to luteolin were measured by using Western blot and fluorometric assay, respectively.Luteolin dose-dependently (10–100 μmol/L) elicited relaxation of PE- or potassium-contracted aortic rings. The vasorelaxation effect of luteolin was attenuated by the eNOS inhibitor, N-nitro-l-arginine methyl ester, suggesting that this luteolin action is at least partially mediated by activating eNOS activity. We further found that luteolin dose-dependently (10–100 μmol/L) increased eNOS phosphorylation at Ser1177 (up to 1.9-fold) in isolated rat rings. Consistently, exposure of HAECs to luteolin also increased eNOS phosphorylation and NO production.Luteolin may be a vascular protective agent by directly acting on vascular ECs to stimulate NO-dependent vascular dilatation.
Keywords: Luteolin; Vasorelaxation, endothelial cells; eNOS; NO

Homocysteine, antioxidant micronutrients and late onset dementia by Lawrence J. Whalley; Susan J. Duthie; Andrew R. Collins; John M. Starr; Ian J. Deary; Helen Lemmon; Ashleigh C. Duthie; Alison D. Murray; Roger T. Staff (277-285).
To distinguish between contributions to dementia made by homocysteine, folate, B12 and antioxidant micronutrients.This is a follow-up study of a sample reported in 2002. Homocysteine was measured at baseline in 201 individuals born in 1921 and without dementia at age 77 years and followed up to age 88 years. Baseline macro- and micronutrient status was estimated from BMI, the MONICA food frequency questionnaire, plasma folate, B12 and, in a subgroup (N = 173), plasma antioxidant micronutrients. Time to dementia onset during follow-up was compared between participants grouped by homocysteine concentration using Cox regression. Model 1 adjusted for age, sex, childhood IQ, education, socioeconomic deprivation, presence of heart disease, hypertension, plasma folate and B12. In model 2 plasma, antioxidants were added to these covariables.During a mean follow-up of about 5 years, there were 39 incident dementia cases among 201 participants. In model 1, being in the highest homocysteine group (>14 μmol/L) was associated with a 234 % increased risk (HR 3.34, 95 % CI 1.16–9.57) of any dementia. After inclusion of plasma antioxidants in model 2, there were 32 incident dementia cases from a subsample (N = 173). Homocysteine >14 μmol was associated with a 272 % increased dementia risk (HR = 3.72, 95 % CI 1.06–13.08).High homocysteine increases the risk of dementia. The association between tHcy and dementia is independent of plasma folate, B12 and antioxidant micronutrient status.
Keywords: Homocysteine; Vitamin B12; Folate; Plasma antioxidants; Dementia

Comparison of cardiovascular aquaporin-1 changes during water restriction between 25- and 50-day-old rats by Vanina A. Netti; Mariana C. Vatrella; Melina F. Chamorro; María I. Rosón; Elsa Zotta; Andrea L. Fellet; Ana M. Balaszczuk (287-295).
Aquaporin-1 (AQP1) is the predominant water channel in the heart, linked to cardiovascular homeostasis. Our aim was to study cardiovascular AQP1 distribution and protein levels during osmotic stress and subsequent hydration during postnatal growth.Rats aged 25 and 50 days were divided in: 3d-WR: water restriction 3 days; 3d-WAL: water ad libitum 3 days; 6d-WR+ORS: water restriction 3 days + oral rehydration solution (ORS) 3 days; and 6d-WAL: water ad libitum 6 days. AQP1 was evaluated by immunohistochemistry and western blot in left ventricle, right atrium and thoracic aorta.Water restriction induced a hypohydration state in both age groups (40 and 25 % loss of body weight in 25- and 50-day-old rats, respectively), reversible with ORS therapy. Cardiac AQP1 was localized in the endocardium and endothelium in both age groups, being evident in cardiomyocytes membrane only in 50-day-old 3d-WR group, which presented increased protein levels of AQP1; no changes were observed in the ventricle of pups. In vascular tissue, AQP1 was present in the smooth muscle of pups; in the oldest group, it was found in the endothelium, increasing after rehydration in smooth muscle. No differences were observed between control groups 3d-WAL and 6d-WAL of both ages.Our findings suggest that cardiovascular AQP1 can be differentially regulated in response to hydration status in vivo, being this response dependent on postnatal growth. The lack of adaptive mechanisms of mature animals in young pups may indicate an important role of this water channel in maintaining fluid balance during hypovolemic state.
Keywords: Aquaporin-1; Dehydration; Heart; Postnatal growth; Vascular tissue

Effects of high-tryptophan diet on pre- and postnatal development in rats: a morphological study by Paola Castrogiovanni; Giuseppe Musumeci; Francesca Maria Trovato; Rosanna Avola; Gaetano Magro; Rosa Imbesi (297-308).
Tryptophan is an essential amino acid, precursor of serotonin. Serotonin (5HT) regulates the secretion of pituitary growth hormone (GH), which in turn stimulates the liver to produce insulin-like growth factor-I (IGF-I) that is necessary for development and growth. The aim of our study was to investigate the effects of an excess of tryptophan in the diet of pregnant rats on the differentiation of skeletal muscle tissue.We conducted an immunohistochemical study on the IGF-I expression in hepatic and muscle tissues in offspring, and then, we associated this molecular data with morphological effects on the structure of the muscle fibers and hepatic tissue at different postnatal weeks, from birth to sexual maturity. Measurements of 5HT, GH in blood, and of tryptophan hydroxylase (Tph) activity in gastrointestinal tracts tissue were also taken.Hyperserotonemia and higher values of Tph activity were detected in both pregnant rats and pups. Very low levels of GH were detected in experimental pups. Morphological alterations of the muscle fibers and lower IGF-I expression in hepatic and muscle tissue in pups were found.Our data suggest that an excess of tryptophan in the diet causes hyperserotonemia in fetus. Hyperserotonemia results in an excess of serotonin in the brain where it has an adverse effect on the development of serotonergic neurons. The affected neurons do not regulate optimally the secretion of pituitary GH that consequently decreases. This limits stimulation in the liver to produce IGF-I, crucial for development and growth of pups.
Keywords: Tryptophan; Serotonin; Growth hormone; Insulin-like growth factor; Development; Diet

Glycaemic index of meals affects appetite sensation but not energy balance in active males by Mei-Yi Wu; Joanna L. Bowtell; Craig A. Williams (309-319).
Foods with low glycaemic index (LGI) are reported to suppress appetite mainly in overweight population but have not been investigated in athletic adults.The aim of this study was to compare the short-term effects of LGI and high GI (HGI) meals over a day on subsequent subjective appetite sensation, energy intake, energy expenditure, energy balance and resting metabolic rate in physically active males.This cross-sectional randomized crossover study included 14 active males (mean ± SD; age 34.5 ± 8.9 years, body mass index 22.8 ± 2.1 kg m−2) to consume LGI and HGI meals on two separate days. On each trial day, participants consumed a breakfast in the laboratory and then left with a packed lunch, dinner and snacks. Appetite scores, energy intake and expenditure were assessed.The area under the curve for appetite scores of the HGI trial was significantly smaller than that of the LGI trial during the laboratory period (p = 0.027) and throughout the day (p = 0.009). No significant differences in energy intake, energy expenditure, energy balance and resting metabolic rate were found between groups, between the trial days and between the corresponding post-trial days.These results show that frequent ingestion of the HGI meals, contrary to the previous reports, suppresses appetite more than that of LGI meals, but did not affect energy balance in physically active normal-weight males.
Keywords: Energy intake; Energy expenditure; Hunger; Satiety; Carbohydrate; Body weight

Positive muscle protein net balance and differential regulation of atrogene expression after resistance exercise and milk protein supplementation by Søren Reitelseder; Jakob Agergaard; Simon Doessing; Ida C. Helmark; Peter Schjerling; Gerrit van Hall; Michael Kjaer; Lars Holm (321-333).
Resistance exercise and amino acid availability are positive regulators of muscle protein net balance (NB). However, anabolic responses to resistance exercise and protein supplementation deserve further elucidation. The purpose was to compare intakes of whey, caseinate (both: 0.30 g/kg lean body mass), or a non-caloric control after heavy resistance exercise on protein turnover and mRNA expressions of forkhead homeobox type O (FOXO) isoforms, muscle RING finger 1 (MuRF1), and Atrogin1 in young healthy males.Protein turnover was determined by stable isotope-labeled leucine and femoral arteriovenous blood samples at rest and during 6-h recovery. Muscle biopsies were collected at −60 min (rest) and at 60, 210, and 360 min in the recovery period.During recovery, leucine NB was significantly higher in the protein groups compared to control (P < 0.001). Differences in leucine NB, rate of disappearance, and oxidation were observed in the early recovery period between whey and caseinate. FOXO1A and MuRF1 were upregulated at 60 and 210 min, and, in contrast, FOXO3 and Atrogin1 were downregulated at 210 and 360 min. For leucine rate of appearance and all FOXO and atrogene mRNA expressions, no differences were observed between groups.Whey and caseinate were equally superior to control in the 6-h recovery period and displayed temporal differences with whey having a fast and superior effect in the early part of the recovery period. Effects on mRNA expressions indicate different regulatory mechanisms on the ubiquitin ligases MuRF1 and Atrogin1 in recovery from heavy resistance exercise.
Keywords: Whey; Caseinate; Muscle protein turnover; Leucine oxidation; Ubiquitin ligases

Determination of anti-inflammatory activities of standardised preparations of plant- and mushroom-based foods by Dhanushka Gunawardena; Kirubakaran Shanmugam; Mitchell Low; Louise Bennett; Suresh Govindaraghavan; Richard Head; Lezanne Ooi; Gerald Münch (335-343).
Chronic inflammatory processes contribute to the pathogenesis of many age-related diseases. In search of anti-inflammatory foods, we have systematically screened a variety of common dietary plants and mushrooms for their anti-inflammatory activity.A selection of 115 samples was prepared by a generic food-compatible processing method involving heating. These products were tested for their anti-inflammatory activity in murine N11 microglia and RAW 264.7 macrophages, using nitric oxide (NO) and tumour necrosis factor-α (TNF-α) as pro-inflammatory readouts.Ten food samples including lime zest, English breakfast tea, honey-brown mushroom, button mushroom, oyster mushroom, cinnamon and cloves inhibited NO production in N11 microglia, with IC50 values below 0.5 mg/ml. The most active samples were onion, oregano and red sweet potato, exhibiting IC50 values below 0.1 mg/ml. When these ten food preparations were retested in RAW 264.7 macrophages, they all inhibited NO production similar to the results obtained in N11 microglia. In addition, English breakfast tea leaves, oyster mushroom, onion, cinnamon and button mushroom preparations suppressed TNF-α production, exhibiting IC50 values below 0.5 mg/ml in RAW 264.7 macrophages.In summary, anti-inflammatory activity in these food samples survived ‘cooking’. Provided that individual bioavailability allows active compounds to reach therapeutic levels in target tissues, these foods may be useful in limiting inflammation in a variety of age-related inflammatory diseases. Furthermore, these foods could be a source for the discovery of novel anti-inflammatory drugs.
Keywords: Food; Inflammation; Cinnamon; Oregano; Oyster mushroom

Soy isoflavones may inhibit tumor cell invasion and metastasis via their effects on matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). The current study investigates the effects of daidzein, R- and S-equol on the invasion of MDA-MB-231 human breast cancer cells and the effects of these compounds on MMP/TIMP expression at the mRNA level.The anti-invasive effects of daidzein, R- and S-equol (0, 2.5, 10, 50 μM) on MDA-MB-231 cells were determined using the Matrigel invasion assay following 48-h exposure. Effects on MMP-2, MMP-9, TIMP-1 and TIMP-2 expression were assessed using real-time PCR. Chiral HPLC analysis was used to determine intracellular concentrations of R- and S-equol.The invasive capacity of MDA-MB-231 cells was significantly reduced (by approximately 50–60 %) following treatment with 50 μM daidzein, R- or S-equol. Anti-invasive effects were also observed with R-equol at 2.5 and 10 μM though overall equipotent effects were induced by all compounds. Inhibition of invasion induced by all three compounds at 50 μM was associated with the down-regulation of MMP-2, while none of the compounds tested significantly affected the expression levels of MMP-9, TIMP-1 or TIMP-2 at this concentration. Following exposure to media containing 50 μM R- or S-equol for 48-h intracellular concentrations of R- and S-equol were 4.38 ± 1.17 and 3.22 ± 0.47 nM, respectively.Daidzein, R- and S-equol inhibit the invasion of MDA-MB-231 human breast cancer cells in part via the down-regulation of MMP-2 expression, with equipotent effects observed for the parent isoflavone daidzein and the equol enantiomers.
Keywords: Daidzein; R-equol; S-equol; Breast cancer; Soy isoflavone