European Journal of Nutrition (v.53, #3)

Dietary fat content is a primary factor associated with the increase in global obesity rates. There is a delay in achieving fat balance following exposure to a high-fat (HF) diet (≥ 40 % of total energy from fat) and fat balance is closely linked to energy balance. Exercise has been shown to improve this rate of adaptation to a HF diet. Recently, however, the role of dietary fatty acid composition on energy and macronutrient balance has come into question. We chose studies that compared monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), and saturated fatty acids (SFA). We have reviewed studies that measured diet-induced thermogenesis (DIT), energy expenditure (EE), or fat oxidation (FOx) in response to a HF meal challenge, or long-term dietary intervention comparing these fatty acids.While single-meal studies show that SFA induce lower DIT and FOx compared to unsaturated fats, the effect of the degree of unsaturation (MUFA vs. PUFA) appears to yet be determined. Long-term dietary interventions also support the notion that unsaturated fats induce greater EE, DIT, and/or FOx versus SFA and that a high MUFA diet induces more weight loss compared to a high SFA diet. Sex and BMI status also affect the metabolic responses to different fatty acids; however, more research in these areas is warranted.SFA are likely more obesigenic than MUFA, and PUFA. The unsaturated fats appear to be more metabolically beneficial, specifically MUFA ≥ PUFA > SFA, as evidenced by the higher DIT and FOx following HF meals or diets.
Keywords: Fatty acids; Saturated fatty acids (SFA); Monounsaturated fatty acids (MUFA); Polyunsaturated fatty acids (PUFA); Energy balance

Maternal diet, bioactive molecules, and exercising as reprogramming tools of metabolic programming by Paulo C. F. Mathias; Ghada Elmhiri; Júlio C. de Oliveira; Carine Delayre-Orthez; Luiz F. Barella; Laize P. Tófolo; Gabriel S. Fabricio; Abalo Chango; Latifa Abdennebi-Najar (711-722).
Nutrition and lifestyle, particularly over-nutrition and lack of exercise, promote the progression and pathogenesis of obesity and metabolic diseases. Nutrition is likely the most important environmental factor that modulates the expression of genes involved in metabolic pathways and a variety of phenotypes associated with obesity and diabetes. During pregnancy, diet is a major factor that influences the organ developmental plasticity of the foetus. Experimental evidence shows that nutritional factors, including energy, fatty acids, protein, micronutrients, and folate, affect various aspects of metabolic programming. Different epigenetic mechanisms that are elicited by bioactive factors in early critical developmental ages affect the susceptibility to several diseases in adulthood. The beneficial effects promoted by exercise training are well recognised, and physical exercise may be considered one of the more prominent non-pharmacological tools that can be used to attenuate metabolic programming and to consequently ameliorate the illness provoked by metabolic diseases and reduce the prevalence of obesity, type 2 diabetes, and cardiovascular diseases. Literature on the different outcomes of unbalanced diets and the beneficial effects of some bioactive molecules during gestation and lactation on the metabolic health of offspring, as well as the potential mechanisms underlying these effects, was reviewed. The importance of the combined effects of functional nutrition and exercise as reprogramming tools of metabolic programming is discussed in depth. Finally, this review provides recommendations to healthcare providers that may aid in the control of early programming in an attempt to optimise the health of the mother and child.
Keywords: Perinatal life; Metabolic programming; Nutrition; Metabolic syndrome; Epigenetic modification; Physical activity

The effect of submicron fat droplets in a drink on satiety, food intake, and cholecystokinin in healthy volunteers by Harry P. F. Peters; Elisabeth C. M. Bouwens; Ewoud A. H. Schuring; Edward Haddeman; Krassimir P. Velikov; Sergey M. Melnikov (723-729).
Small fat droplets infused into the gut reduce food intake and hunger more than bigger ones, at levels as low as 6 g, and these effects are hypothesized to occur via satiety hormones such as cholecystokinin. It is, however, unknown whether the effect of droplet size would persist after oral consumption. It is also unknown whether an even smaller droplet size can affect hunger and food intake and at what minimum amount of fat. Therefore, the aim of the study was to test the effect of very fine fat droplets on satiety and food intake in two different quantities.In a balanced-order 4-way crossover design, 24 volunteers consumed a fat-free meal replacement drink with either 5 or 9 g oil (rapeseed) and either 3 or 0.1 μm droplet size. Appetite scores and plasma cholecystokinin levels (in n = 12 subset) were measured for 180 min, when food intake was assessed during an ad libitum meal. Data were analyzed by ANCOVA, followed by Dunnett’s test and paired t test. The behavior of the emulsions was also characterized in a simulated gastrointestinal model.Despite faster in vitro lipolysis of the smallest droplets, neither droplet size nor fat amount affected satiety or food intake. From t = 45–150 min, cholecystokinin response was 50 % higher (P < 0.05) after the 0.1 versus 3 μm, but only with 9 g fat.When this particular fat at these amounts is delivered in a meal replacement drink, droplet size does not influence appetite or food intake. This effect is independent of the amount of fat or plasma cholecystokinin changes.
Keywords: Satiety; Food intake; Cholecystokinin; Lipids; Droplet size

Dietary, lifestyle, and genetic determinants of vitamin D status: a cross-sectional analysis from the European Prospective Investigation into Cancer and Nutrition (EPIC)-Germany study by Tilman Kühn; Rudolf Kaaks; Birgit Teucher; Frank Hirche; Jutta Dierkes; Cornelia Weikert; Verena Katzke; Heiner Boeing; Gabriele I. Stangl; Brian Buijsse (731-741).
Considerable variation in 25-hydroxyvitamin D (25(OH)D) in populations worldwide that seems to be independent of latitude has been reported. Therefore, we aimed to assess vitamin D status of a mid-aged German general population and to identify its dietary, lifestyle, anthropometric, and genetic determinants. 25(OH)D concentrations were measured by LC–MS/MS in plasma samples of a random subcohort of the German arm of the European Prospective Investigation into Cancer and Nutrition (EPIC) comprising 2,100 subjects aged 35–65 years. Associations between potential predictors and 25(OH)D were assessed by linear regression models.32.8 % of the variance in 25(OH)D was explained by a multivariable regression model, with season being the by far strongest predictor (semi-partial R 2: 14.6 %). Sex, waist circumference, leisure time physical activity, smoking, polymorphisms in the GC, CYP2R1, and DHCR7 genes, supplement use, exogenous hormone use, alcohol consumption, egg consumption, and fish consumption were significantly associated with 25(OH)D concentrations as well. However, none of these factors explained >2.3 % of the variance in 25(OH)D.Even with a comprehensive set of genetic, anthropometric, dietary, and lifestyle correlates, not more than 32.8 % of the variation in 25(OH)D could be explained in the EPIC-Germany study, implying that vitamin D prediction scores may not provide an appropriate proxy for measured 25(OH)D. Food intake was only a weak predictor of 25(OH)D concentrations, while a strong seasonal fluctuation in 25(OH)D was shown.
Keywords: Vitamin D; 25-Hydroxyvitamin D; Diet; Lifestyle; Single nucleotide polymorphisms; Prediction model

The principal function of the adipose tissue is the storage of energy in the form of triglyceride through the process of adipogenesis, as well as the provision of the stored energy through lipolysis. In the present study, we investigated the effect of hydroxytyrosol on lipolysis in 3T3-L1 adipocytes.3T3-L1 adipocytes, used as in vitro model in this study, were treated with several concentration of hydroxytyrosol. Glycerol release was measured to identify the lipolytic rate activation. All factors activation and expression were carried out via Western blotting and qRT-PCR.Our results showed that hydroxytyrosol, over a range of concentrations, attenuated triglyceride accumulation and stimulated glycerol release in fully differentiated adipocytes in a dose- and time-dependent manner. Moreover, hydroxytyrosol had no effect on adipocyte viability. To understand the mechanism underlying hydroxytyrosol-stimulated lipolysis, we used inhibitors of PKA, PKC, PKG, ERK1/2, and nitric oxide production. Pretreatment with a PKA inhibitor (Rp-cAMPs) and an ERK1/2 inhibitor (U0126) significantly attenuated hydroxytyrosol-stimulated lipolysis. In contrast, a PKC inhibitor (Calphostin C), 2 PKG inhibitors (KT 5823 and Rp-cGMPs), and a nitric oxide inhibitor (S-ethyl ITU) had no effect on hydroxytyrosol-stimulated lipolysis. Over the same range of concentrations, hydroxytyrosol downregulated the expression of adipose triglyceride lipase, hormone sensitive lipase (HSL), and adipogenesis-related transcription factors PPARγ and C/EBPα. In addition, hydroxytyrosol increased the phosphorylation rate of HSL at Ser563 and Ser660, as well as perilipin and ERK phosphorylation.Hydroxytyrosol induced lipolysis in 3T3-L1 adipocytes via the activation of PKA and ERK1/2 pathway.
Keywords: Hydroxytyrosol; 3T3-L1; Lipolysis; PKA; ERK1/2; ATGL; HSL; Perilipin

High-dose cholecalciferol supplementation significantly increases peripheral CD4+ Tregs in healthy adults without negatively affecting the frequency of other immune cells by Barbara Prietl; Gerlies Treiber; Julia K. Mader; Evelyne Hoeller; Michael Wolf; Stefan Pilz; Winfried B. Graninger; Barbara M. Obermayer-Pietsch; Thomas R. Pieber (751-759).
Regulatory T cells (Tregs) play a central role in the maintenance of self-tolerance. Animal and in vitro studies suggest that vitamin D is involved in reducing the risk of autoimmunity by modulating Tregs.In a double-blind, placebo controlled study in 60 healthy volunteers, we assessed the effect of a 12-week high-dose oral cholecalciferol supplementation (140,000 IU/month) on the number and function of CD4posCD25highFoxP3posCD127dim Tregs. We also assessed the clinical safety of the supplementation and the effect on the frequency of other immune cells such as monocytes, dendritic cells, natural killer cells, natural killer T cells, B cells and subgroups of T cells. We also tested the in vitro effect of cholecalciferol on Tregs in human cell cultures.By using FACS analysis, ex vivo suppressive co-cultures and apoptosis assays, we were able to show that a cholecalciferol supplementation leads to significantly increased numbers of peripheral Tregs in vivo. Tregs function and the frequency of other immune cells remained unchanged, and no clinically relevant safety concerns were found. The in vitro exposure of human peripheral blood mononuclear cells to cholecalciferol also supported our in vivo findings.Our results indicate a substantial effect of a supplementation with inactive vitamin D on the immune system of healthy humans in vivo and provide a rationale for future studies to investigate the immunomodulatory effects of vitamin D in autoimmune diseases.
Keywords: Vitamin D; Regulatory T cells; Immune regulation; Immunotherapy; Tolerance

Dietary conjugated α-linolenic acid did not improve glucose tolerance in a neonatal pig model by Christian-Alexandre Castellano; Jean-Patrice Baillargeon; Mélanie Plourde; Sandie I. Briand; Paul Angers; Alain Giguère; J. Jacques Matte (761-768).
There is an increased interest in the benefits of conjugated α-linolenic acid (CLNA) on obesity-related complications such as insulin resistance and diabetes. The aim of the study was to investigate whether a 1 % dietary supplementation of mono-CLNA isomers (c9-t11-c15-18:3 + c9-t13-c15-18:3) improved glucose and lipid metabolism in neonatal pigs.Since mono-CLNA isomers combine one conjugated two-double-bond system with an n-3 polyunsaturated fatty acid (PUFA) structure, the experimental protocol was designed to isolate the dietary structural characteristics of the molecules by comparing a CLNA diet with three other dietary fats: (1) conjugated linoleic acid (c9-t11-18:2 + t10-c12-18:2; CLA), (2) non-conjugated n-3 PUFA, and (3) n-6 PUFA. Thirty-two piglets weaned at 3 weeks of age were distributed among the four dietary groups. Diets were isoenergetic and food intake was controlled by a gastric tube. After 2 weeks of supplementation, gastro-enteral (OGTT) and parenteral (IVGTT) glucose tolerance tests were conducted.Dietary supplementation with mono-CLNA did not modify body weight/fat or blood lipid profiles (p > 0.82 and p > 0.57, respectively) compared with other dietary groups. Plasma glucose, insulin, and C-peptide responses to OGTT and IVGTT in the CLNA group were not different from the three other dietary groups (p > 0.18 and p > 0.15, respectively). Compared to the non-conjugated n-3 PUFA diet, CLNA-fed animals had decreased liver composition in three n-3 fatty acids (18:3n-3; 20:3n-3; 22:5n-3; p < 0.001).These results suggest that providing 1 % mono-CLNA is not effective in improving insulin sensitivity in neonatal pigs.
Keywords: Conjugated linolenic acid; n-3 fatty acid; Insulin resistance; Pig

Low dietary calcium and obesity: a comparative study in genetically obese and normal rats during early growth by Clarisa Marotte; Gabriel Bryk; Macarena M. S. Gonzales Chaves; Fima Lifshitz; Maria Luz Pita Martín de Portela; Susana N. Zeni (769-778).
A low calcium intake (LCaI) may predispose to obesity, and excessive fat mass may be detrimental to bone. The impact of Ca inadequacy would be greater in subjects predisposed to obesity. LCaI effect on obesity development during the rapid growth period was compared in two strains of rats: spontaneously obese IIMb/β (O) and Wistar (W). Pregnant rats were fed 0.5 % (N) or 0.2 % (L) of Ca (OLCa, ONCa, WLCa and WNCa). Male pups were fed the maternal diet until day 60. Body composition, lipid profile, glucose homeostasis, 25 hydroxyvitamin D, Ca-phosphorus, and bone metabolism were evaluated.BW and body fat were higher, whereas body protein was lower in OLCa versus ONCa (p < 0.05). OLCa presented the highest body fat, glucose, non-HDL and total cholesterol, TGL, insulin levels, and HOMA-IR, liver weight, and adipose perigonadal plus retroperitoneal pads (p < 0.05). WLCa did not exhibit an increase BW and only showed a slight change in body composition with minor biochemical alterations compared to WNCa (p < 0.05). Osteocalcin, CTX, and proximal tibia and lumbar spine BMDs were lower in O than in W rats fed the same Ca diet (p < 0.05). Body ash and Ca content, and total skeleton BMC/BW were lower in OLCa and WLCa versus their corresponding NCa groups (p < 0.05).The negative effect of a low Ca diet on fat mass accumulation and lipid profile may be more evident in rats predisposed to obesity. Nevertheless, low CaI interferes with the normal glucose homeostasis leading to an increase in insulin resistance. Low CaI during early growth may be an obesogenic factor that may persist into adult life and may account for the development of obesity and some of its co-morbidities.
Keywords: Low calcium diet; Obesity; Osteocalcin; Insulin resistance; Bone remodeling; Bone mass

Fucoxanthin in association with Vitamin c acts as modulators of human neutrophil function by A. C. Morandi; N. Molina; B. A. Guerra; A. P. Bolin; R. Otton (779-792).
Neutrophils provide the first line of defense of the innate immune system by phagocytosing, killing and digesting bacteria and fungi. During this process, neutrophils produce reactive oxygen species (ROS), which in excess, can damage the cells themselves and surrounding tissues. The carotenoid fucoxanthin (Fc) has been studied concerning its antioxidant and anti-inflammatory actions. Vitamin c (Vc) also demonstrates potent antioxidant action. This study aimed to evaluate the effect of Fc (2 μM) in association with Vc (100 μM) on functional parameters of human neutrophils in vitro.We evaluated the migration and phagocytic capacity, intracellular calcium mobilization, ROS production (O 2 ·− , H2O2, HOCl), myeloperoxidase activity, profile of antioxidant enzymes, phosphorylation of p38 MAPK and p65 NFκB subunit, GSH/GSSG ratio and release of pro-inflammatory cytokines (TNF-α and IL-6) in neutrophils under different stimuli.We verified an increase in phagocytic capacity for all treatments, together with an increase in intracellular calcium only in cells treated with Fc and Fc + Vc. ROS production was reduced by all treatments, although Vc was a better antioxidant than Fc. Phosphorylation of the p-65 subunit of NFκB was reduced in cells treated with Fc + Vc and release of TNF-α and IL-6 was reduced by all treatments. These findings indicate that the regulation of inflammatory cytokines by neutrophils is not exclusively under the control of the NFκB pathway. Fc reduced the activity of some antioxidant enzymes, whereas Vc increased GR activity and the GSH/GSSG ratio.In conclusion, the results presented in this study clearly show an immunomodulatory effect of the carotenoid fc alone or in combination with Vc on the function of human neutrophils.
Keywords: Carotenoids; Antioxidant; Anti-inflammatory; Leukocytes; Free radical

Consumption of wheat bran modified by autoclaving reduces fat mass in hamsters by Scott V. Harding; Harry D. Sapirstein; Todd C. Rideout; Christopher P. F. Marinangeli; Arshala K. M. Dona; Peter J. H. Jones (793-802).
To investigate the effect that wheat bran modified by autoclaving (MWB) had on reducing fat accumulation in hamsters fed a hypercholesterolemia- and obesity-inducing diet.Male hamsters (n = 45) were randomized into 3 groups and fed a hypercholesterolemia- and obesity-inducing diet with or without 10 % standard wheat bran or MWB for 28 days. Our outcome measures included body composition measured by DXA, oxygen consumption and plasma lipids and glucose concentrations.Animals fed the MWB diet had lower % fat mass (49.8 vs. 53.4 %; p = 0.02) and higher % lean body mass (47.2 vs. 44.1 %; p = 0.02) compared with controls despite no differences in food intake or weight gain. Additionally, plasma glucose tended to be lower (6.9 vs. 8.5 mmol/l; p < 0.08) in the MWB animals compared with controls.Our data suggest that the compositional changes in autoclaved wheat bran, specifically solubility of phenolic antioxidants and fiber, may have contributed to the lower fat accumulation in our animals. Further study is needed to determine whether the exact mechanism involved increased lipolysis and energy utilization from adipose.
Keywords: Adiposity; Wheat bran; Weight management; Dual-energy X-ray absorptiometry

The impact of protein supplementation on cognitive performance in frail elderly by Nikita L. van der Zwaluw; Ondine van de Rest; Michael Tieland; Jos J. Adam; Gert Jan Hiddink; Luc J. C. van Loon; Lisette C. P. G. M. de Groot (803-812).
Maintenance of cognitive abilities is important for elderly to stay independent. With the aging of the population, the call for modifiable factors is emerging. Dietary protein might improve cognitive performance; however, this has hardly been studied. Therefore, we studied the impact of 24-week dietary protein supplementation on cognitive performance in pre-frail and frail elderly people.Pre-frail and frail elderly subjects, according to the Fried criteria, randomly received a protein drink containing 15 g protein or a placebo drink twice a day. Cognitive performance was measured at baseline and after 24 weeks by means of a sensitive neuropsychological test battery. In addition, reaction time was assessed after both 12 and 24 weeks of intervention. Domain scores were calculated for the domains episodic memory, attention and working memory, information processing speed, and executive functioning. Analyses of covariance were used to determine differences between groups. Linear mixed models were used to determine differences in reaction time over time and per treatment.In total, 65 subjects (79 ± 8 years) with a median Mini-Mental State Examination score of 28 (interquartile range 26–30) were included. Reaction time improved more in the protein group (68 ms) than in the placebo group (18 ms, P = 0.03). Dietary protein had no significant effect on any of the cognitive domain scores.Protein supplementation might improve reaction time performance in pre-frail and frail elderly, but did not improve other cognitive functions.
Keywords: Nutrition; Protein supplementation; Frail elderly; Cognitive performance; Aging

Long-term dietary l-arginine supplementation increases endothelial nitric oxide synthase and vasoactive intestinal peptide immunoexpression in rat small intestine by Ksenija Velickovic; Milica Markelic; Igor Golic; Vesna Otasevic; Ana Stancic; Aleksandra Jankovic; Milica Vucetic; Biljana Buzadzic; Bato Korac; Aleksandra Korac (813-821).
Nitric oxide (NO) and vasoactive intestinal polypeptide (VIP) are important intestinal neurotransmitters that coexist in the gut enteric nervous system and play an important role in intestinal physiology (e.g., absorption, motility, fluid secretion and smooth muscle relaxation). It is also known that cold exposure alters several aspects of gastrointestinal physiology and induces hyperphagia to meet increased metabolic demands, but there are no data regarding NO and VIP involvement in intestinal response during acclimation to cold. The objective of this study was to determine the influence of long-term l-arginine supplementation on the expression of the three isoforms of nitric oxide synthase (NOS) and VIP in small intestine of rats acclimated to room temperature or cold.Animals (six per group) acclimated to room temperature (22 ± 1 °C) and cold (4 ± 1 °C), respectively, were treated with 2.25 % l-arginine, a substrate for NOSs, or with 0.01 % N ω-nitro-l-arginine methyl ester, an inhibitor of NOSs, for 45 days. The topographical distribution of VIP and NOSs expression in small intestine was studied by immunohistochemistry, and ImageJ software was used for semiquantitative densitometric analysis of their immunoexpression.Long-term dietary l-arginine supplementation increases VIP and NOSs immunoexpression at room temperature while at cold increases the endothelial NOS, inducible NOS and VIP but decrease neuronal NOS in rat small intestine.Our results demonstrate that long-term dietary l-arginine supplementation modulates NOSs and VIP immunoexpression in rat small intestine with respect to ambient temperature, pointing out the eNOS as a predominant NOS isoform with an immunoexpression pattern similar to VIP.
Keywords: Arginine; Small intestine; Nitric oxide synthase; Vasoactive intestinal peptide

Oxidative stress and inflammation in obesity after taurine supplementation: a double-blind, placebo-controlled study by Flávia Troncon Rosa; Ellen Cristini Freitas; Rafael Deminice; Alceu Afonso Jordão; Julio Sérgio Marchini (823-830).
Some researchers found decreased levels of plasma taurine in obese subjects and animals, and reduced expression of an important enzyme of taurine synthesis. These evidences, coupled with the metabolic imbalance of obesity and the possible anti-inflammatory and antioxidant effects of taurine, highlighted the use of taurine as a supplement in obesity treatment. The aim of the present study was to investigate whether taurine supplementation, associated with nutritional counseling, modulates oxidative stress, inflammatory response, and glucose homeostasis in obese women.A randomized double-blind placebo-controlled study was conducted with 16 women with obesity diagnosis and 8 women in the normal weight range. The obese volunteers were matched by age and body mass index and randomly assigned to either the placebo (3 g/day starch flour) or taurine (3 g/day taurine) group. The study lasted 8 weeks, and the experimental protocol included nutritional assessment and determination of plasma sulfur amino acids, insulin, and adiponectin, serum glycemia, and markers of inflammatory response and oxidative stress.Plasma taurine levels were significantly decreased (41 %) in the obese volunteers. Both the placebo and taurine groups showed significant reduction in weight (3 %), with no differences between groups. Different from placebo, taurine-supplemented group showed significant increase in plasma taurine (97 %) and adiponectin (12 %) and significant reduction in the inflammatory marker hs-C-reactive protein (29 %) and in the lipid peroxidation marker thiobarbituric acid reactive substances (TBARS) (20 %).Eight weeks of taurine supplementation associated with nutritional counseling is able to increase adiponectin levels and to decrease markers of inflammation (high-sensitivity C-reactive protein) and lipid peroxidation (TBARS) in obese women.
Keywords: Taurine; Obesity; Inflammation; Oxidative stress; Adiponectin

Plantago maxima leaves extract inhibits adipogenic action of a high-fat diet in female Wistar rats by Alexey A. Tinkov; Olga N. Nemereshina; Elizaveta V. Popova; Valentina S. Polyakova; Viktor A. Gritsenko; Alexandr A. Nikonorov (831-842).
The primary objective of this study is to investigate the content of biologically active compounds producing an antioxidant effect in Plantago maxima and their influence on main mechanisms of dietary obesity development.Biologically active compounds in P. maxima were tested using paper chromatography. In in vivo experiment, high-fat-fed Wistar rats obtained P. maxima water extract for 3 months. Morphometric parameters, weight gain, serum adipokines, and cytokines, as well as oxidative stress biomarkers in rats’ tissues were evaluated. Gut microflora was also examined. Plantago maxima leaves used in the experiment contained significant amount of flavonoids, iridoids, phenol carboxylic acids, and tannins and ascorbic acid. Our in vivo experiment data demonstrate that P. maxima water extract prevents excessive adiposity in a diet-induced model. P. maxima consumption reduced serum leptin (twofold), macrophage chemoattractant protein-1 (sevenfold), tumor-necrosis factor-α (25 %), and interleukine-6 (26 %) levels. P. maxima water extract decreased adipose tissue oxidative stress biomarkers in rats fed a high-fat diet. In addition, increased bacterial growth in the diet-induced obesity model was reversed by the P. maxima extract treatment. Plantago maxima water extract possessed antiadipogenic, antidiabetic, antiinflammatory, antioxidant activity, and normalized gut microflora in a rat model of diet-induced excessive adiposity due to a high content of biologically active compounds.
Keywords: Plantago maxima ; Phytochemicals; Excessive adiposity; Inflammation; Oxidative stress; Endocrine dysfunction

d,l-Sulforaphane (SFN) is a promising chemopreventive agent with in vivo efficacy against prostate cancer in experimental rodents. This study was undertaken to determine the role of vimentin and plasminogen activator inhibitor-1 (PAI-1) in anticancer effects of SFN.Effect of SFN on levels of different proteins was determined by Western blotting or immunofluorescence microscopy. RNA interference of vimentin and PAI-1 was achieved by transient transfection. Apoptosis was quantified by flow cytometry. Transwell chambers were used to determine cell migration.Exposure of PC-3 and DU145 human prostate cancer cells to SFN resulted in induction of vimentin protein, which was accompanied by down-regulation of E-cadherin protein expression. The SFN-mediated induction of vimentin was also observed in a normal human prostate epithelial cell line. RNA interference of vimentin did not have any appreciable effect on early or late apoptosis resulting from SFN exposure. On the other hand, SFN-mediated inhibition of PC-3 and DU145 cell migration was significantly augmented by knockdown of the vimentin protein. Knockdown of vimentin itself was inhibitory against cell migration. The SFN-treated cells also exhibited induction of PAI-1, which is an endogenous inhibitor of urokinase-type plasminogen activator system. Similar to vimentin, PAI-1 knockdown resulted in a modest augmentation of PC-3 cell migration inhibition by SFN. Tumors from SFN-treated transgenic adenocarcinoma of mouse prostate mice showed a 1.7-fold increase in vimentin protein level compared with control tumors.The present study indicates that vimentin and PAI-1 inductions confer modest protection against SFN-mediated inhibition of prostate cancer cell migration.
Keywords: Sulforaphane; Vimentin; Prostate cancer; Chemoprevention

Phase-II metabolism limits the antiproliferative activity of urolithins in human colon cancer cells by Antonio González-Sarrías; Juan Antonio Giménez-Bastida; María Ángeles Núñez-Sánchez; Mar Larrosa; María Teresa García-Conesa; Francisco A. Tomás-Barberán; Juan Carlos Espín (853-864).
Urolithins, gut microbiota metabolites derived from ellagic acid and ellagitannins, reach micromolar concentrations in the colon lumen where can have anti-inflammatory and anticancer effects. The antiproliferative activity of urolithins (Uro-A, Uro-B, Uro-C and Uro-D) and their most relevant in vivo glucuronides were evaluated in three human colon cancer cell lines (Caco-2, SW480 and HT-29).Cell proliferation was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide and Trypan blue exclusion assays. Cell cycle was evaluated by flow cytometry and urolithins metabolism by HPLC–MS/MS.Urolithins inhibited cell proliferation and cell cycle progression in a time- and dose-dependent manner and arrested the cells at S and G2/M phases, depending on the urolithin. Uro-A exerted the highest antiproliferative activity, followed by Uro-C, Uro-D and Uro-B. Unlike Caco-2 and SW480 cells, HT-29 cells partially overcame the effects after 48 h, which was related to the complete glucuronidation of urolithins. Uro-A or Uro-B glucuronides did not affect cell cycle and showed lower antiproliferative activity than their aglycone counterparts. Uro-A or Uro-B plus inhibitors of drug efflux ABC transporters partially prevented the glucuronidation of urolithins in HT-29 cells which became more sensitive.Uro-A, Uro-B, Uro-C and Uro-D exerted different antiproliferative effects depending on the colon cancer cell line. We also report here, for the first time, the role of ABC transporters and Phase-II metabolism in HT-29 cells as a mechanism of cancer resistance against urolithins due to their conversion to glucuronide conjugates that exerted lower antiproliferative activity.
Keywords: Urolithins; Ellagic acid; Glucuronide; Cell cycle; Colon cancer; Phase-II metabolism

Pediatric adiposity stabilized in Switzerland between 1999 and 2012 by Stefanie B. Murer; Siret Saarsalu; Michael B. Zimmermann; Isabelle Aeberli (865-875).
Several countries have recently reported stabilization and/or a decrease in the prevalence of pediatric obesity. However, systematic, repeated national monitoring studies are scarce, and it is unclear whether this trend would be sustained. The objective was to present the latest overweight and obesity prevalence in Swiss children and to investigate trends in prevalence from 1999 to 2012.Using probability-proportionate-to-size cluster sampling, nationally representative samples of children aged 6–12 years were recruited in 1999 (n = 594), 2002 (n = 2,493), 2004 (n = 328), 2007 (n = 2,218), 2009 (n = 907), and 2012 (n = 2,963). Height and weight were measured to calculate BMI (kg/m2). BMI cutoffs proposed by the Centers for Disease Control and Prevention (CDC) and by the International Obesity Task Force were used to determine the prevalence of overweight (excluding obesity) and obesity. Waist circumference was measured in 2007 and 2012, and multiple skinfold thicknesses assessed in 2002 and 2012.Using the CDC criteria, prevalences of overweight and obesity in 2012 were 11.9 % (95 % CI 10.7–13.1) and 7.1 % (95 % CI 6.2–8.0), respectively, and did not change between 1999 and 2012 (β = −0.144, p = 0.293 and β = −0.063, p = 0.552, respectively). Boys had significantly higher obesity prevalence than girls in 2007 (5.6 vs. 3.4 %) and 2012 (8.1 vs. 5.9 %). Percentage of children with excess waist circumference and body fat percentage did not differ between 2007 and 2012, and 2002 and 2012, respectively.Our data indicate the prevalence of childhood adiposity in Switzerland stabilized between 1999 and 2012, but ≈1 in 5 children remain overweight or obese and further efforts are needed to control the epidemic.
Keywords: Obesity; Overweight; Children; Trends; Body fat percentage; Waist circumference

Anti-anaemia efficacy of β-lactoglobulin hydrolysate-iron complex on iron-deficient anaemic rats by Jie Zhou; Xue-Ying Mao; Xu Wang; Ting Ai; Jing-Jiao Ma; Ying-Hui Li (877-884).
The effects of orally administered β-lactoglobulin hydrolysate-iron complex (β-LGH-Fe) on haematological and biochemical parameters in anaemic rats were evaluated. Female weaning Sprague–Dawley rats were fed with iron-deficient diet to induce iron deficiency anaemia. After 6 weeks, the obtained anaemic rats were divided into five groups: iron deficiency control group (iron-deficient diet without β-LGH-Fe complex supplementation, IDC); three groups supplemented with different dosages of β-LGH-Fe complex (0.5 mg Fe/kg BW, iron-deficient diet with low β-LGH-Fe, IDLFe; 2.0 mg Fe/kg BW, iron-deficient diet with medium β-LGH-Fe, IDMF; 4.0 mg Fe/kg BW, iron-deficient diet with high β-LGH-Fe, IDHFe); and ferrous sulphate-supplemented group at a dosage of 2.0 mg Fe/kg BW.β-LGH-Fe complex could significantly improve hematocrit and haemoglobin decrease, and normalise the serum iron level, total iron-binding capacity and transferrin saturation of anaemic rats in a dose-dependent manner. Serum ferritin content and hepatic nonheme iron level were also increased. In addition, the antioxidant enzyme activities of superoxidase dismutase, catalase and glutathione peroxidase in both plasma and liver homogenate were improved. The production of malondialdehyde and pro-inflammatory cytokines (TNF-α and IL-6) decreased.It suggests that β-LGH-Fe complex can ameliorate iron deficiency anaemia, which might make it a potential ingredient with anti-anaemia activity.
Keywords: β-Lactoglobulin hydrolysate-iron complex; Haematological parameters; Serum ferritin; Serum transferrin

Diet-induced obese rats have higher iron requirements and are more vulnerable to iron deficiency by Jesse Bertinato; Cristina Aroche; Louise J. Plouffe; Megan Lee; Zehra Murtaza; Laura Kenney; Christopher Lavergne; Alfred Aziz (885-895).
Since obesity is associated with poorer iron status, the effects of diet-induced obesity on iron status and iron-regulatory pathways were examined.Weanling male diet-induced obese sensitive (n = 12/diet group) and resistant (n = 12/diet group) rats were fed one of four high-fat, high-energy diets supplemented with 5 (5Fe, low), 15 (15Fe, marginal), 35 (35Fe, normal) or 70 (70Fe, high) mg iron/kg diet for 12 weeks. At the end of the study, rats in each diet group were categorised as obese (>19 %) or lean (<17 %) based on percentage body fat.Obese rats gained more weight, had larger total lean mass, consumed more food and showed greater feed efficiency compared with lean rats. Obese rats fed the 5Fe and 15Fe diets had poorer iron status than lean rats fed the same diet. Obese 5Fe rats had lower serum iron and more severe iron-deficiency anaemia. Obese 15Fe rats had lower mean corpuscular haemoglobin and liver iron concentrations. Hepcidin mRNA expression in liver and adipose tissue was similar for obese and lean rats. Iron concentration and content of the iron transporters divalent metal transporter 1 and ferroportin 1 in duodenal mucosa were also similar.Obese rats that were larger, regardless of adiposity, had higher iron requirements compared with lean rats that appeared independent of hepcidin, inflammation and intestinal iron absorption. Higher iron requirements may have resulted from larger accretion of body mass and blood volume. Greater food consumption did not compensate for the higher iron needs, indicating increased susceptibility to iron deficiency.
Keywords: Anaemia; Diet-induced obesity; Hepcidin; Iron deficiency; Rat

Molecular pathology of acute kidney injury in a choline-deficient model and fish oil protective effect by Valeria Denninghoff; Georgina Ossani; Ana Uceda; Matias Rugnone; Elmer Fernández; Cristóbal Fresno; German González; Maria Luisa Díaz; Alejandra Avagnina; Boris Elsner; Alberto Monserrat (897-906).
The aim of this work was to investigate the potential protective effects of fish oil on the basis of kidney transcriptomic data on a nutritional experimental model.Male weanling Wistar rats were divided into four groups and fed choline-deficient (CD) and choline-supplemented (CS) diets with vegetable oil (VO) and menhaden oil (MO): CSVO, CDVO, CSMO and CDMO. Animals were killed after receiving the diets for 6 days. Total RNA was purified from the right kidney and hybridized to Affymetrix GeneChip Rat Gene 1.0 ST Array. Differentially expressed genes were analyzed.All CSVO, CSMO and CDMO rats showed no renal alterations, while all CDVO rats showed renal cortical necrosis. A thorough analysis of the differential expression between groups CSMO and CDMO was carried out. There were no differential genes for p < 0.01. The analysis of the differential expression between groups CSVO and CSMO revealed 32 genes, 11 were over-expressed and 21 were under-expressed in CSMO rats.This work was part of a large set of experiments and was used in a hypothesis-generating manner. The comprehensive analysis of genetic expression allowed confirming that menhaden oil has a protective effect on this nutritional experimental model and identifying 32 genes that could be responsible for that protection, including Gstp1. These results reveal that gene changes could play a role in renal injury.
Keywords: Acute kidney injury; Choline-deficient; Gene expression; Menhaden oil; Protective effect

Hepatic lipid overloading induces lipotoxicity which can cause hepatocyte damage, fibrosis, and eventually progress to cirrhosis, which is associated with nonalcoholic fatty liver disease. Adiponectin receptors play important roles in regulating lipid metabolism. In this study, we used a lentivirus system to overexpress the adiponectin receptor 1 (AdipoR1) in HepG2 cells to define the role of adiponectin and its receptor 1 in the development of fatty liver syndrome. Exposure of human hepatocytes, HepG2 cells, to palmitate (0.2 or 0.4 mM) for 16 h resulted in elevated apoptosis, whereas AdipoR1 decreased the palmitate-induced apoptosis. Transgene AdipoR1 increased the expression of FATP2, acyl-coA oxidase, and carnitine palmitoyltransferase I in palmitate-treated HepG2 cells. The transcript level of acetyl-CoA carboxylase and fatty acid synthase was upregulated by palmitate treatment, while AdipoR1 reversed the effect induced by palmitate. AdipoR1 increased the gene expression of cytochrome C oxidase, peroxisome proliferator-activated receptor α, and decreased the gene expression of PGC1α and AMPKα in HepG2 cells under palmitate treatment. Palmitate suppressed ATP production, while transgene AdipoR1 reversed the decreased ATP production by palmitate. Transgene AdipoR1 enhanced AKT phosphorylation in HepG2 cells both with and without palmitate treatment. When PI3 kinase inhibitor was applied, the protective effect of AdipoR1 was absent, such that palmitate again decreased ATP production while also reducing cell viability.AdipoR1 enhances fatty acid metabolism and cell viability in palmitate-treated HepG2 cells partially by activating AKT signaling. Therefore, AdipoR1 has therapeutic potential in the treatment of nonalcoholic fatty liver disease.
Keywords: Adiponectin receptor 1; HepG2; Apoptosis; Lipid metabolism; PI3K/AKT

Protective effects of garlic extract on cardiac function, heart rate variability, and cardiac mitochondria in obese insulin-resistant rats by Luerat Supakul; Hiranya Pintana; Nattayaporn Apaijai; Siriporn Chattipakorn; Krekwit Shinlapawittayatorn; Nipon Chattipakorn (919-928).
Garlic has been shown to exhibit antioxidant effects and cardioprotective properties. However, the effects of garlic extract on the heart in insulin resistance induced by long-term high-fat-diet consumption are not well defined. Therefore, we sought to determine the effects of garlic extract in the obese insulin-resistant rats.Male Wistar rats (180–200 g) were divided into two groups: normal-diet or high-fat-diet (n = 24/group) fed for 12 weeks. Rats in each groups were divided into three subgroups (n = 8 each): vehicle or garlic extract (250 or 500 mg/kg/day, respectively) treated for 28 days. At the end of the treatment, the metabolic parameters, heart rate variability (HRV), cardiac function, and cardiac mitochondrial function were determined.Rats that received a high-fat-diet for 12 weeks had increased body weight, visceral fat, plasma insulin levels, total cholesterol, oxidative stress levels, depressed HRV, and cardiac mitochondrial dysfunction. Garlic extract at both concentrations significantly decreased the plasma insulin, total cholesterol, homeostasis model assessment index, and oxidative stress levels. Furthermore, garlic extract at both doses restored the HRV, cardiac function, and cardiac mitochondrial function.We concluded that garlic extract at both concentrations exerted cardioprotective effects against cardiac dysfunction and mitochondrial dysfunction in obese insulin-resistant rats.
Keywords: Garlic extract; High-fat-diet; Insulin resistance; Cardiac function; Mitochondrial function

Apoptosis is a major cause of myocyte death, and taurine is anti-apoptotic. Heat shock protein 70 (HSP70) (which is regulated by heat shock factor—HSF-1) is also anti-apoptotic, and caspase 3 stimulates the apoptotic pathway. This study investigated whether taurine affects atherogenic diet-induced myocardial apoptosis, and whether HSP70, HSF-1 and caspase 3 are involved.New Zealand white rabbits were divided into 3 groups for 4 weeks according to their diet. Group 1 (control) was fed a normal rabbit diet; Group 2 (MC) received a normal rabbit diet with 1 % methionine plus 0.5 % cholesterol. Group 3 received MC diet + 2.5 % taurine (MCT).The atherogenic diet did not affect myocardial HSP70 or HSF-1 protein, but increased myocardial apoptotic nuclei to 40 % (p < 0.01) versus 7 % in con and 12 % in MCT (p < 0.01). However, in MCT, myocardial HSP70 expression increased by 42.7 % versus con and MC (p = 0.016), HSF-1 by 12 % versus con and MC (p < 0.05), and total nuclei count increased by 37 % versus MC (p < 0.05). Caspase 3 subunits remained unchanged in all groups, and HSP70 was increased approximately twofold in endothelial layer of arterioles (p = 0.01).This study shows that taurine could reduce myocardial apoptotic nuclei and thus confer myocardial cytoprotection via stimulating myocardial HSP70 via HSF-1 and caspase 3-independent mechanisms.
Keywords: Cholesterol; Homocysteine; Methionine; Taurine; Hypochlorite

Meal replacement based on Human Ration modulates metabolic risk factors during body weight loss: a randomized controlled trial by Natalia Elizabeth Galdino Alves; Bárbara Nery Enes; Hércia Stampini Duarte Martino; Rita de Cássia Gonçalves Alfenas; Sônia Machado Rocha Ribeiro (939-950).
A meal replacement may be an effective strategy in the management of obesity to increase antioxidant intake, attenuating oxidative stress and inflammation. In the present study, we investigated the efficacy of a new nutritional supplement to reduce metabolic risk parameters in obese women.In a randomized controlled crossover study (2 × 2), 22 women (percentage body fat 40.52 ± 3.75 %; body mass index—BMI 28.72 ± 2.87 kg/m2; 35.04 ± 5.6 years old) were allocated into two treatments: hypocaloric diet and drink containing “Human Ration” (HR) consumption (CRHR), and hypocaloric diet and control drink consumption (CR). The study consisted of 2 periods of 5 weeks with 1 week of washout in two orders (CR → CRHR and CRHR → CR). Caloric restriction was 15 %, based on estimated energy requirement. Anthropometric, clinical and metabolic risk parameters were assessed at baseline and at the end of each period.Some metabolic risk factors were favorably modulated in both interventions: reduction in body weight (CR −0.74 ± 1.27 kg; p = 0.01; CRHR −0.77 ± 1.3 kg; p = 0.02), body mass index (BMI) (CR −0.27 ± 0.51 kg/m2; p = 0.02; CRHR −0.30 ± 0.52 kg/m2; p = 0.01) and HOMA-IR (CR −0.35 ± 0.82; p = 0.02, CRHR −0.41 ± 0.83; p = 0.03). However, CRHR reduced waist circumference (−2.54 ± 2.74 cm; p < 0.01) and gynoid fat (−0.264 ± 0.28 g; p < 0.01), and increased HDL-c levels (0.08 ± 0.15 mmol/l; p = 0.04).Associated with hypocaloric diet, the intake of a nutritional supplement rich in phytochemicals as a breakfast substitute for 5 weeks had no additional effect on weight reduction than caloric restriction alone, but increased central lipolysis and improved the lipoprotein profile.
Keywords: Obesity management; Meal replacements; Phytochemicals; Oxidative stress; Low-grade inflammation; Metabolic risk

TNFα blockade for inflammatory rheumatic diseases is associated with a significant gain in android fat mass and has varying effects on adipokines: a 2-year prospective study by Éric Toussirot; Laurent Mourot; Barbara Dehecq; Daniel Wendling; Émilie Grandclément; Gilles Dumoulin (951-961).
To evaluate the long-term consequences of TNFα inhibitors on body composition and fat distribution, as well as changes in serum adipokines in patients with rheumatoid arthritis (RA) or ankylosing spondylitis (AS).Eight patients with RA and twelve with AS requiring a TNFα inhibitor were prospectively followed for 2 years. Body composition was evaluated by dual X-ray absorptiometry and included measurements of total fat mass, lean mass, fat in the gynoid and android regions, and visceral fat. Serum leptin, total and high molecular weight (HMW) adiponectin, resistin, and ghrelin were also assessed.There was a significant gain in body mass index (p = 0.05) and a tendency for weight (p = 0.07), android fat (p = 0.07), and visceral fat (p = 0.059) increase in patients with RA, while in AS, total fat mass significantly increased (p = 0.02) with a parallel weight gain (p = 0.07). When examining the whole population of patients, we observed after 2 years a significant increase in body weight (+1.9 %; p = 0.003), body mass index (+2.5 %; p = 0.004), total fat mass (+11.1 %; p = 0.007), and fat in the android region (+18.3 %; p = 0.02). There was a substantial, albeit nonsignificant gain in visceral fat (+24.3 %; p = 0.088). Lean mass and gynoid fat were not modified. No major changes were observed for serum leptin, total adiponectin, and ghrelin, while HMW adiponectin and the HMW/total adiponectin ratio tended to decrease (−15.2 %, p = 0.057 and −9.3 %, p = 0.067, respectively). Resistin decreased significantly (−22.4 %, p = 0.01).Long-term TNFα inhibition in RA and AS is associated with a significant gain in fat mass, with a shift to the android (visceral) region. This fat redistribution raises questions about its influence on the cardiovascular profile of patients receiving these treatments.
Keywords: TNFα inhibitors; Body composition; Fat mass; Cardiovascular risk; Adipokines; Resistin

Long-term cysteine fortification impacts cysteine/glutathione homeostasis and food intake in ageing rats by Karine Vidal; Denis Breuillé; Patrick Serrant; Philippe Denis; Françoise Glomot; Fabienne Béchereau; Isabelle Papet (963-971).
Healthy ageing is associated with higher levels of glutathione. The study aimed to determine whether long-term dietary fortification with cysteine increases cysteine and glutathione pools, thus alleviating age-associated low-grade inflammation and resulting in global physiological benefits.The effect of a 14-week dietary fortification with cysteine was studied in non-inflamed (NI, healthy at baseline) and in spontaneously age-related low-grade inflamed (LGI, prefrail at baseline) 21-month-old rats. Fifty-seven NI rats and 14 LGI rats received cysteine-supplemented diet (4.0 g/kg of free cysteine added to the standard diet containing 2.8 g/kg cysteine). Fifty-six NI rats and 16 LGI rats received a control alanine-supplemented diet.Cysteine fortification in NI rats increased free cysteine (P < 0.0001) and glutathione (P < 0.03) in the liver and the small intestine. In LGI rats, cysteine fortification increased total non-protein cysteine (P < 0.0007) and free cysteine (P < 0.03) in plasma, and free cysteine (P < 0.02) and glutathione (P < 0.01) in liver. Food intake decreased over time in alanine-fed rats (r 2 = 0.73, P = 0.0002), whereas it was constant in cysteine-fed rats (r 2 = 0.02, P = 0.68). Cysteine fortification did not affect inflammatory markers, mortality, body weight loss, or tissue masses.Doubling the dietary intake of cysteine in old rats increased cysteine and glutathione pools in selected tissues. Additionally, it alleviated the age-related decline in food intake. Further validation of these effects in the elderly population suffering from age-related anorexia would suggest a useful therapeutic approach to the problem.
Keywords: Ageing rats; Dietary cysteine; Frailty; Low-grade inflammation

Red grape berry-cultured cells reduce blood pressure in rats with metabolic-like syndrome by A. Leibowitz; Z. Faltin; A. Perl; Y. Eshdat; Y. Hagay; E. Peleg; E. Grossman (973-980).
Cumulative evidence suggests that moderate red wine consumption protects the cardiovascular system. The effect of cultured cells derived from red grape berry (RGC) on blood pressure (BP) has not been investigated. We therefore studied the antihypertensive effects of oral consumption of RGC in experimental rat model of metabolic-like syndrome and assessed its effect on human umbilical vein endothelial cells (HUVECs).Forty male Sprague–Dawley rats were fed for 5 weeks with either a high fructose diet (HFD) (n = 10) or HFD supplemented, during the last 2 weeks, with different doses (200, 400 and 800 mg/kg/day) of RGC suspended in their food (n = 30). BP, plasma triglycerides, insulin and adiponectin levels were measured at the beginning and after 3 and 5 weeks of diet. RGC effect on vasodilatation was evaluated by its ability to affect endothelin-1 (ET-1) production and endothelial nitric oxide synthase (eNOS) expression in HUVECs.BP, plasma triglycerides, insulin and adiponectin increased significantly in rats fed with a HFD. The increase in BP, plasma triglycerides and insulin was attenuated by RGC supplementation. Incubation of HUVECs with RGC demonstrated a concentration-dependent inhibition of ET-1 secretion and increase in the level of eNOS, signaling a positive effect of RGC on vasodilatation.In rats with metabolic-like syndrome, RGC decreased BP and improved metabolic parameters. These beneficial effects may be mediated by the cell constituents, highly rich with polyphenols and resveratrol, reside in their natural state.
Keywords: Rats; Metabolic syndrome; Blood pressure; Red grape-cultured cell; Endothelin-1; Endothelial NO synthase

Plasma kinetics of chylomicron-like emulsion and lipid transfers to high-density lipoprotein (HDL) in lacto-ovo vegetarian and in omnivorous subjects by Juliana C. Vinagre; Carmen C. G. Vinagre; Fernanda S. Pozzi; Cristiane Z. Zácari; Raul C. Maranhão (981-987).
Previously, it was showed that vegan diet improves the metabolism of triglyceride-rich lipoproteins by increasing the plasma clearance of atherogenic remnants. The aim of the current study was to investigate this metabolism in lacto-ovo vegetarians whose diet is less strict, allowing the ingestion of eggs and milk. Transfer of lipids to HDL, an important step in HDL metabolism, was tested in vitro.Eighteen lacto-ovo vegetarians and 29 omnivorous subjects, all eutrophic and normolipidemic, were intravenously injected with triglyceride-rich emulsions labeled with 14C-cholesterol oleate and 3H-triolein. Fractional clearance rates (FCR, in min−1) were calculated from samples collected during 60 min. Lipid transfer to HDL was assayed by incubating plasma samples with a donor nanoemulsion labeled with radioactive lipids.LDL cholesterol was lower in vegetarians than in omnivores (2.1 ± 0.8 and 2.7 ± 0.7 mmol/L, respectively, p < 0.05), but HDL cholesterol and triglycerides were equal. Cholesteryl ester FCR was greater in vegetarians than in omnivores (0.016 ± 0.012, 0.003 ± 0.003, p < 0.01), whereas triglyceride FCR was equal. Cholesteryl ester transfer to HDL was lower in vegetarians than in omnivores (2.7 ± 0.6, 3.5 ± 1.5 %, p < 0.05), but free cholesterol, triglyceride and phospholipid transfers and HDL size were equal.Similarly to vegans, lacto-ovo vegetarian diet increases remnant removal, as indicated by cholesteryl oleate FCR, which may favor atherosclerosis prevention, and has the ability to change lipid transfer to HDL.
Keywords: Triglycerides; Emulsions; Nanoparticles; Vegetarian diets; Lipoprotein metabolism; Cholesterol

Incorporation of conjugated linoleic acid isomers into porcine erythrocytes by Tomaž Malovrh; Enver Melkić; Drago Kompan; Alenka Levart; Lidija Kompan (989-993).
The aim of the current study was to determine the incorporation of cis (c) 9, trans (t) 11-conjugated linoleic acid (CLA) and t10, c12-CLA into porcine erythrocytes—both isomers were supplemented in equal proportions.The study group consisted of 16 piglets randomly assigned into experimental and control group. For the period of 5 weeks, the piglets from the experimental group were receiving a 1.2 % CLA supplement while the controls were supplemented with the same amount of sunflower oil. For the remaining 7 weeks, the piglets were fed without a supplement. Blood samples to evaluate incorporation of CLA into erythrocyte membranes were taken from all animals on weekly basis.Compared to t10, c12-CLA isomer, proportion of c9, t11-CLA isomer in the membrane of erythrocytes was higher for the whole time of the study period. After 4 weeks of feeding, it approaches the plateau. The peak value for both isomers was measured at the end of week 5, with a value of 3.24 g c9, t11-CLA/100 g of fatty acids and a 1.09 g t10, c12-CLA/100 g of fatty acids (p < 0.0001). After cessation of supplementation, the proportion of both isomers gradually decreased to be almost completely washed out—in 7 weeks.During supplementation with equivalent amounts of CLA isomers, their proportion in membranes of porcine erythrocytes increases with time, with higher proportion of c9, t11-CLA. CLA isomers probably differently incorporate into different cell membranes at different species which could explain its various biological functions.
Keywords: Conjugated linoleic acid; Porcine erythrocytes; Fatty acid methyl esters