European Journal of Nutrition (v.50, #6)

Markers of metabolic syndrome in obese children before and after 1-year lifestyle intervention program by C. Pedrosa; B. M. P. M. Oliveira; I. Albuquerque; C. Simões-Pereira; M. D. Vaz-de-Almeida; F. Correia (391-400).
Excess weight may be related to the development of adverse cardiometabolic risk factors in children. The aim of this study was to evaluate the effect of a lifestyle intervention program (nutrition and exercise counseling) on anthropometric parameters and metabolic syndrome (MS) components in Portuguese overweight/obese children.A total of 83 overweight/obese children aged 7–9 years were assigned to a 1-year individual or group-based treatment (GT); 61 children (z-score BMI (zBMI): 1.93 ± 0.28; 27 boys and 34 girls) completed the program. Anthropometric and biochemical parameters were assessed at baseline, at 6 months and at 1 year.The overweight/obese children, compared to normal-weight ones, presented significantly higher blood pressure, total-cholesterol, total-cholesterol/high density lipoprotein cholesterol (HDL) ratio, triglycerides, Apolipoprotein B and C-reactive protein levels, while HDL and Apolipoprotein A-I were significantly lower. At baseline, the prevalence of MS was 16.4% in overweight/obese and 0% in normal-weight children. The number of components of MS was significantly higher in children with higher zBMI. Lifestyle intervention led to a significant improvement in zBMI, waist circumference/height ratio, HDL, triglycerides, Apolipoprotein A-I, and Apolipoprotein B levels. The prevalence of MS decreased to 14.8%. The GT intervention seems to be more successful, with a significant decrease in zBMI and an increase in HDL and a lower drop-out rate.Overweight/obese children have multiple risk factors associated with the MS. Lifestyle intervention, both individual and group-based treatment, led to an improvement in the degree of overweight/obesity and in MS components.
Keywords: Children; Metabolic syndrome; Obesity; Nutrition; Lifestyle intervention

Grape seed extract ameliorates tumor necrosis factor-α-induced inflammatory status of human umbilical vein endothelial cells by Chia-Lun Chao; Nen-Chung Chang; Ching-Sung Weng; Kueir-Rarn Lee; Shung-Te Kao; Jiin-Chyr Hsu; Feng-Ming Ho (401-409).
Inflammation has played a key role in the causation of atherosclerosis. However, the effects of grape seed extract (GSE) on the pro-inflammatory intracellular signaling, enzyme activity, and inflammatory mediators of endothelial cells have not been sufficiently studied, and less information exists on the comparison between GSE and vitamin C, a well-known antioxidant compound, on their anti-inflammatory properties.We investigated the effects of GSE and vitamin C on the cell viability, oxidative stress, monocyte adhesion, the expression of nuclear factor-κB inhibitor (IκB), intercellular adhesion molecule-1 (ICAM-1) and cyclooxygenase-2 (COX-2), and the production of prostaglandin E2 (PG E2) in TNF-α-treated human umbilical vein endothelial cells (HUVECs).Cell viability was measured by MTT assay. The adhesion of THP-1 to HUVECs was evaluated by cell adhesion assay. The oxidized nucleoside 8-hydroxydeoxyguanosine (8-OHdG) (an indicator of oxidative damage to DNA), ICAM-1, and PG E2 were measured by ELISA. IκB and COX-2 expression were evaluated by western blot analysis.TNF-α (10, 20, and 50 ng/mL), GSE (50 and 200 μg/mL), or vitamin C (100 μM) did not affect cell viability. GSE (50–100 μg/mL) attenuated TNF-α (20 ng/mL)-induced 8-OHdG production, THP-1 adhesion, the expression of IκB degradation, ICAM-1 and COX-2, and the production of PGE2 in a dose-dependent manner. Vitamin C (100 μM) also showed significant antioxidative and anti-inflammatory effects.GSE effectively ameliorates TNF-α-induced inflammatory status of HUVECs. The findings of the present study suggest that consumption of GSE may be beneficial to inflammatory atherosclerosis.
Keywords: Inflammation; Grape seed extract; Adhesion; Cyclooxygenase-2; Prostaglandin E2 ; Endothelial cell

To assess the effect of medium-chain fatty acid (MCFA)-rich mustard oil and polyunsaturated fatty acid (PUFA)-rich mustard oil on erythrocyte membrane composition and osmotic fragility in normal and hypercholesterolemic rats.Membrane composition was analyzed using standard kits. Osmotic fragility was determined using method described by Dacie and Lewis. Fatty acid composition of membrane was analyzed using gas chromatographic methods. Membrane shape analysis was performed using scanning electron microscope.Osmotic fragility data suggested that the erythrocyte membrane of hypercholesterolemic rats were relatively more fragile than that of the normal rat’s membrane, which could be reversed with the addition of MCFA- and PUFA-rich oil in the diet. The increased plasma cholesterol in hypercholesterolemic rats could also be lowered by the experimental oils. There was also marked changes in the fatty acid composition of the plasma and erythrocyte membrane phospholipids. Polyunsaturated fatty acids decreased in the plasma of the hypercholesterolemic subjects were increased with the treatment of the experimental oils. Shape changes of the membrane holes were observed in the hypercholesterolemic condition, which was brought to normal shape with the administration of the experimental oils.In conclusion, rat erythrocytes appear to be deformed and became more fragile in cholesterol-rich blood. This deformity and fragility was partially reversed by experimental oils by virtue of their ability to lower the extent of hypercholesterolemia.
Keywords: Hypercholesterolemia; Erythrocyte membrane; Membrane fragility; Membrane lipid profile; Scanning electron microscopy

Dietary deoxynucleic acid induces type 2 T-helper immune response through toll-like receptor 9 in mice by Mariko Nakamoto; Emi Shuto; Toshio Hosaka; Tohru Sakai (421-426).
It has been shown that dietary nucleotides modulate immune response. Due to their unique properties in immune responses, nucleotides are used as immunonutrition in the field of clinical nutrition.In this study, we examined the effect of dietary deoxynucleic acid (DNA) on antigen (Ag)-specific immune response in ovalbumin (OVA)-immunized BALB/c mice and determined the mechanism using toll-like receptor 9 (TLR9) knock-out (KO) mice.BALB/c or TLR9 KO mice were fed control and 1% DNA diets and immunized with OVA. Spleen cells from OVA-immunized mice were stimulated with OVA in vitro, and the contents of IFN-γ and IL-4 in supernatants were measured by an ex vivo system. CD11c+ dendritic cells were purified, and ability of cytokine induction to CD4+ cells was examined.The level of OVA-specific IL-4 production in the DNA group was significantly higher than that in the control group. In contrast, the level of OVA-specific IFN-γ production in the DNA group was lower than that in the control group. The DNA diet decreased Ag-specific IL-4 production and enhanced Ag-specific IFN-γ production in TLR9 KO mice. CD11c+ DCs from mice fed the DNA diet had a greater ability than CD11c+ DCs from mice fed the control diet to induce the production of IL-4 from DO11.10 CD4+ T cells.Dietary DNA increases Ag-specific IL-4 production and decreases IFN-γ production through a TLR9-dependent pathway. CD11c+ dendritic cells are target cells in dietary DNA-induced immune regulation.
Keywords: Nucleic acid; T-helper; Dendritic cell; IL-4; CD4+ T cell

Lymphatic absorption of choline plasmalogen is much higher than that of ethanolamine plasmalogen in rats by Megumi Nishimukai; Maya Yamashita; Yudai Watanabe; Yuya Yamazaki; Toru Nezu; Ryouta Maeba; Hiroshi Hara (427-436).
Plasmalogen is a subclass of phospholipids widely distributed in animal tissues and ingested as food; however, the absorptive characteristics of different classes of plasmalogen have not been clarified.Our object was to compare the lymphatic output of choline and ethanolamine plasmalogens after an administration of phospholipid preparations containing each class of plasmalogens, and to analyze molecular species of plasmalogen absorbed into the lymph.A duodenal infusion of 1 ml of 10% emulsion of choline phospholipid (PC) containing 50.6% choline plasmalogen (PlsCho) or ethanolamine phospholipid (PE) containing 52.5% ethanolamine plasmalogen (PlsEtn) was administered in the lymph duct-cannulated rats. Molecular species of plasmalogen absorbed into the lymph were measured by LC-MS/MS.Lymph outputs of PlsCho and PlsEtn increased and reached a peak value at 3 h after PC and PE injection, respectively. The peak value of PlsCho was much higher and remained at a high level until 8 h, whereas PlsEtn output fell to half of the peak value at 7 h. Total lymphatic output of PlsCho was 5-times higher than that of PlsEtn. Compositions of sn-1 in lymph plasmalogens roughly reflected those of the injected lipids, whereas sn-2 in both PlsCho and PlsEtn was rich in arachidonic acid (20:4) regardless of the composition of the administered fatty acid. Both plasmalogen and lysoplasmalogen after PE injection were not released into the portal vein.Lymphatic absorption of PlsCho is much higher than that of PlsEtn in rats, and plasmalogens are re-esterified as 20:4-rich forms in the small intestine
Keywords: Plasmalogen; Phospholipids; UPLC-MS/MS; Lymph absorption

Vitamin C levels in blood are influenced by polymorphisms in glutathione S-transferases by Alexandra Horska; Csilla Mislanova; Stefano Bonassi; Marcello Ceppi; Katarina Volkovova; Maria Dusinska (437-446).
Glutathione S-transferases (GSTs) are intimately involved in combating oxidative stress and in detoxifying xenobiotics. Our objective was to examine possible interactions between polymorphisms in GST genes and plasma vitamin C, tocopherols and carotenoids in 149 reference subjects and 239 subjects occupationally exposed to mineral fibres (asbestos, rock wool, glass fibre), agents that induce oxidative stress.Deletion of GSTM1 and GSTT1, and substitution 105Ile/Val in GSTP1 genes were determined by PCR, antioxidants in plasma were measured by HPLC.Tocopherols and carotenoids were affected by age, sex, smoking, occupational exposure to fibres, but not by GST polymorphisms. Vitamin C level was influenced by sex, smoking and occupational exposure. Subjects with deletion of GST had lower vitamin C levels compared with subjects carrying the functional gene variant. Vitamin C levels varied according to GSTM1 polymorphism in the whole group (p < 0.05), in all reference subjects (p < 0.05), in the asbestos factory reference group (p < 0.05), and according to GSTT1 polymorphism in reference group of the rock wool plant (p < 0.05). Vitamin C levels were approximately 20% lower in subjects with both functionally deficient genes in the whole group (p < 0.01) and in all non-exposed subjects (p < 0.05).The correspondence of lower vitamin C levels with non-functional GST isoenzymes may indicate a causal connection between two antioxidant defence pathways, also the underlying mechanism is not yet clear. It seems that supplementation by natural antioxidants is particularly important for subjects with unfavourable genetic makeup and in those exposed to oxidative stress.
Keywords: GST polymorphisms; Antioxidants; Vitamin C; Oxidative stress; Exposure to mineral fibres; Individual susceptibility

l-Arabinose uncompetitively inhibits intestinal sucrase by forming an enzyme-inhibitor-substrate (EIS) complex. The transient period of the EIS complex affects the time span of inhibition. We determined the apparent transient period of the EIS complex of sucrase, l-arabinose, and sucrose both in vitro and in humans.Intestinal acetone powder (a source of sucrase), l-arabinose, and sucrose were mixed and injected into a dialysis membrane that was placed in a sucrose solution. The production rate of d-glucose and the release rate of l-arabinose from sucrase were determined. We also investigated the suppression of blood glucose levels by l-arabinose in 21 healthy volunteers. Sucrose (40 g) was ingested with or without l-arabinose (2 g), then blood glucose values were measured, which returned to steady-state conditions within 2 h. Volunteers were then given 90 g of commercial adzuki bean jelly containing 40 g sucrose as the sucrose load, and blood glucose values were measured again.Addition of l-arabinose reduced the production rate of d-glucose compared to the rates measured in the absence of l-arabinose for several hours in vitro. l-Arabinose was released at a lower rate in the presence of sucrose than in its absence. Blood glucose values measured 2 h after sucrose was given with l-arabinose were significantly lower than those measured when l-arabinose was not given (Δ change in maximum value: with l-arabinose, 53.8 ± 19.7 mg/dL; without l-arabinose, 65.0 ± 17.7 mg/dL).The EIS complex of sucrase-l-arabinose-sucrose was maintained for several hours both in vitro and in humans.
Keywords: l-Arabinose; EIS complex; Transient period; Second meal effect

Short-term isocaloric manipulation of carbohydrate intake: effect on subsequent ad libitum energy intake by Adela Penesova; Colleen A. Venti; Joy C. Bunt; Susan M. Bonfiglio; Susanne B. Votruba; Jonathan Krakoff (455-463).
Isocaloric manipulation of carbohydrate or fat intake could alter subsequent ad libitum food intake.In a controlled inpatient study, we investigated whether isocaloric manipulation of carbohydrate or fat would alter subsequent ad libitum energy intake. Eighteen non-diabetic subjects (age range 19–53 years.; 15 M/3F; % body fat 38.5 ± 9.1 (mean ± SD)) were fed for 3 days an isocaloric high-carbohydrate diet (HC; 60% carbohydrate, 20% fat, 20% protein) and a high-fat diet (HF; 50% fat, 30% carbohydrate, 20% protein) in random order each followed by 3 days of ad libitum food intake.There were no differences in mean daily energy intake (EI) following each diet (HC vs. HF: 4,811 ± 1,190 vs. 4,823 ± 1,238 kcal/d; P = 0.7) or in the percent of weight maintenance energy needs (%EN-WM; 173 ± 41 vs. 173 ± 46%, P = 0.5). However, the individual difference in EI between the HF versus HC diet (ΔEI) both on day one and over the 3 days of each ad libitum period was negatively associated with % body fat (%BF) and waist circumference (day 1: ΔEI vs. %BF, r = −0.49, P = 0.04; mean day 1–3 kcal ΔEI vs. %BF, r = −0.66, P = 0.003, and ΔEI vs. waist, r = −0.65, P = 0.004).A short-term isocaloric HC diet did not result in overall lower EI compared with a HF diet in the same individuals. However, we did find that increasing body fat was associated with less decline in EI following the HC versus HF diet indicating that increasing adiposity is associated with altered regulation of EI in response to macronutrient changes.
Keywords: Carbohydrates; Adiposity; Energy intake; High-fat diet; Clinical nutrition

Epidermal growth factor receptor (EGFR/ErbB1) is a transmembrane protein with tyrosine kinase activity activated mainly by ligand, EGF. Matrix metalloproteinases (MMPs) are a family of proteinases that catalyses the destruction of ECM, among which MMP-9 has important role in tumor cell invasion. Secretion of MMP-9 is stimulated by a variety of factors, EGFR being significant. Epigallocatechin-3-gallate (EGCG) is a major polyphenol of green tea that inhibits cell proliferation and invasion. Here, we study the effect of EGFR alone and in collaboration with fibronectin on the status of MMP-9 in human breast cancer cell MDA-MB-231 and its molecular mechanism; study the role of EGCG on the induced MMP-9; and elucidate the signaling molecules involved in the process.We performed zymography, immunoblots, real-time RT-PCR, cell adhesion assay, siRNA studies, and electrophoretic mobility shift assay to demonstrate the findings.EGF induces MMP-9 activity and expression; FAK, PI3 K, and ERK are mainly involved in the process. EGF also causes the transactivation of MMP-9 gene by increasing the DNA binding activity of the transcription factors. EGCG downregulates EGF-induced MMP-9 expression by inhibiting the involved regulatory kinases. EGF collaborates with fibronectin to create a synergistic response, and EGCG inhibits the synergistic response in MDA-MB-231.The study demonstrates the requirement of cross talk between cell matrix adhesion molecules and growth factor receptors to improve biological responses and shows FAK/ERK as the pivotal point of this convergence in human breast carcinoma cell line MDA-MB-231. We also establish EGCG as the potential anti-tumor agent in human breast carcinoma.
Keywords: Epidermal growth factor receptor; MMP-9; Fibronectin; EGCG; Breast cancer; Signaling

Leucine is suggested to act as nutrient signal of high-protein diets regulating pathways associated with an alleviation of metabolic syndrome parameters. However, the subject remains controversial.The aim of this study was to assess and to compare the effects of high-protein diets with dietary leucine supplementation in mice, particularly on energy homeostasis, body composition, and expression of uncoupling protein (UCP), which are suggested to decrease food energy efficiency.Male C57BL/6 mice were exposed for 14 weeks to semi-synthetic diets containing either 20% (adequate protein content, AP) or 50% whey protein (high-protein content, HP). A third group was fed the AP diet supplemented with L-leucine (AP + L) corresponding to the leucine content of the HP diet. The total fat content was 5% (w/w).Body weight gain, body composition, energy expenditure, and protein expression of UCP1 in brown adipose tissue, and UCP3 in skeletal muscle were not different between groups. In HP-fed mice, a stronger increase in blood glucose levels was detected during glucose tolerance tests compared to AP and AP + L, whereas plasma insulin was similar in all groups. Leucine supplementation did not affect glucose tolerance. Plasma cholesterol was significantly decreased in HP and AP + L when compared to AP. Plasma triglyceride concentrations were increased twofold in HP-fed mice when compared to AP + L and AP groups. Liver and skeletal muscle triglyceride and glycogen concentrations were similar in all groups. Postabsorptive plasma concentrations of branched-chain amino acids were not significantly increased after exposure to HP and AP + L diets, whereas those of lysine were decreased in HP and AP + L mice when compared to AP (P < 0.001). Plasma methionine concentrations were lower after HP intake when compared to AP and AP + L (P < 0.05).We suggest that an exposure of mice to HP diets or a corresponding leucine supplementation has no significant effect on energy homeostasis and UCP expression compared with AP diets when feeding a low-fat diet. The use of high-quality whey protein might at least in part explain the results obtained.
Keywords: Dietary protein; Leucine supplementation; Energy expenditure; Body composition; Uncoupling protein; Mice