European Journal of Nutrition (v.48, #1)

The effect of marine n-3 fatty acids in different doses on plasma concentrations of Lp-PLA2 in healthy adults by Maria Weinkouff Pedersen; Prof. Wolfgang Koenig; Prof. Jeppe Hagstrup Christensen; Prof. Erik Berg Schmidt (1-5).
Lipoprotein-associated phospholipase A2 (Lp-PLA2) is an emerging independent risk factor for cardiovascular disease (CVD). Lp-PLA2 can be modified by lipid lowering drugs, but it is unknown whether diet can reduce plasma levels of Lp-PLA2.The aim of the trial was to study the effect of marine n-3 polyunsaturated fatty acids (PUFA) on plasma Lp-PLA2 levels in healthy subjects.Sixty healthy subjects were randomized to a moderate dose (2 g) of n-3 PUFA, a high dose (6.6 g) of n-3 PUFA or olive oil (control) daily for 12 weeks. Plasma Lp-PLA2 was measured at baseline and after the interventions.Plasma Lp-PLA2 levels were unchanged in all three groups before and after the supplements. Neither did the results differ between groups. There was no correlation between the content of n-3 PUFA in platelets or granulocytes or plasma Lp-PLA2.Marine n-3 PUFA had no effect on plasma levels of Lp-PLA2 in healthy adults and relatively young people.
Keywords: omega-3 fatty acids; coronary heart disease; Lp-PLA2 ; fish

Foreword (1-1).

Effects of high-AGE beverage on RAGE and VEGF expressions in the liver and kidneys by Takashi Sato; Xuegang Wu; Noriko Shimogaito; Jun-ichi Takino; Sho-ichi Yamagishi; Masayoshi Takeuchi (6-11).
The formation and accumulation of advanced glycation end products (AGEs) increase in some lifestyle-related diseases as well as in aging; however, little is known about the relationship between food-derived AGEs and the pathology of such diseases.To explore whether food items containing high levels of AGEs are involved in the development of lifestyle-related diseases, rats were orally administered a commercial high-AGE beverage [Lactobacillus beverage-A (LB-A)]. With a particular focus on angiogenesis-associated diseases, the gene expressions of vascular endothelial growth factor (VEGF) and the receptor for AGEs (RAGE) were examined in the liver and kidneys using real-time reverse transcription-polymerase chain reaction. Moreover, AGE deposition was immunohistochemically investigated in these tissues.Hepatic VEGF expression was significantly increased in rats administered LB-A (P < 0.01 vs. control). Furthermore, immunohistochemical analysis detected glucose-derived AGE-positive cells in the liver from the LB-A group. These results suggest that AGE-rich beverages increase hepatic VEGF expression and AGE accumulation, bringing about early events associated with lifestyle-related diseases.
Keywords: Lactobacillus beverage; food-derived AGEs; lifestyle-related diseases; VEGF; RAGE

Enteral and parenteral nutrition distinctively modulate intestinal permeability and T cell function in vitro by Claudia Guzy; Anja Schirbel; Daniela Paclik; Bertram Wiedenmann; Axel Dignass; Andreas Sturm (12-21).
Nutritional support is an established element of therapy for various indications. However, its impact on the mucosal barrier function is not well understood.We investigated the influence of EN and PN on intestinal epithelial cells and peripheral blood (PBMC) and lamina propria mononuclear cells (LPMC), all of which are involved in the mucosal defense against bacterial translocation and systemic inflammation.Integrity of epithelial cells was measured as transepithelial electrical resistance (TER) of confluent Caco-2 monolayers in the presence of 1% EN, PN and a parenteral amino acid mixture (AM). To determine wound healing capacities, an established migration model with IEC-6 cells was used. Furthermore, we investigated apoptosis, cell activation, proliferation and cytokine secretion of Caco-2, HT29 and of stimulated PBMC and LPMC cultured with or without 1 and 5% EN, AM or PN.We demonstrated that EN, AM and PN promoted the integrity of the epithelial monolayer and reconstituted epithelial cell continuity TGF-β-dependently and -independently. Interestingly, only PN induced apoptosis and decreased the mitochondrial membrane potential. The activation status of PBMC was significantly reduced by EN and AM. Specifically, EN leads to an increased apoptosis rate, inhibited cell cycle progression and increased pro-inflammatory cytokine secretion. Both EN and PN reduced the activation status and the release of pro- and anti-inflammatory cytokines.Our study provides evidence that by promoting wound healing and regulating T cell function, EN, AM, and PN potently interact with the intestinal barrier and immune system, thus justifying its use in diseases accompanied by impaired mucosal barrier function.
Keywords: enteral nutrition; epithelial cells; T cells; intestinal permeability; inflammation

Within the framework of Codex Alimentarius, attempts are being made at international level to establish guidelines for use of nutrition and health claims. An important issue that has to be addressed is the process of scientific substantiating of claims on foods.To provide an insight into the current step procedure of the proposed draft recommendations on the scientific basis of health claims. These Codex recommendations are intended to facilitate governments’ own evaluation of health claims made by the industry.Review of comments of governments, observers and non-governmental organizations (NGOs) and relevant references to the proposed draft recommendations of the last sessions of the Codex Committee on Nutrition and Food for Special Dietary Uses (CCNFSDU). A literature search was performed using the PubMed database.Several proposed draft recommendations on the scientific substantiation of health claims have been considered and amended by the CCNFSDU in recent years but the work is not yet complete. The current work draws on the work of FUFOSE and PASSCLAIM and also on that of WHO and FDA. Given the important role of Codex in food safety, the draft recommendations emphasize circumstances where additional evaluation of safety or nutritional safety needs to be considered. High quality human intervention studies are the prime evidence needed to substantiate claims but there is recognition that, in some cases, only observational studies may be available. Animal and in vitro studies will also be evaluated as part of the totality of the evidence. It has been suggested that the recommendations should include re-evaluation of claims after a certain time period, or if new evidence calls into question the scientific validity underpinning the claims. Setting out a common approach for the substantiation of health claims is an important step in the use of health claims around the world. There is a need to reflect emerging as well as consensus science. The substantiating evidence should be proportionate to the claim. Further progress in the elaboration of this relevant Codex text is needed to reach consensus.
Keywords: Guidance for governments; Scientific substantiation; Health claims; Totality of evidence

Previous human studies on the effect of dietary calcium supplementation on faecal excretion of bile acids (BA) and faecal water concentrations of animal neutral sterols (NSt, cholesterol and its metabolites) lack detailed information about single BA and NSt.We investigated whether single BA and NSt in faeces and especially in faecal water are affected by calcium supplementation and whether this affects genotoxicity of faecal water. In addition, we differentiated between men and women with regard to the concentrations of BA and NSt in faecal water.Thirty-one healthy volunteers consumed a calcium supplemented bread (1.0 g/day) and a placebo bread, respectively, for 4 weeks in a double-blind, randomised cross-over trial. Faeces were collected quantitatively for 5 days in the last week of each period. NSt and BA were analysed by GC–MS.Due to calcium supplementation faecal concentrations of lithocholic acid (LCA, 14%, P = 0.008), deoxycholic acid (DCA, 19%, P < 0.001) and 12keto-deoxycholic acid (12keto DCA, 29%, P = 0.049) significantly increased whereas BA concentrations in faecal water were only marginally affected. In contrast, concentrations of cholesterol (30%, P = 0.020) and its metabolites coprostanol (43%, P = 0.004), coprostanone (36%, P = 0.003), cholestanol (44%, P = 0.001) and cholestenone (32%, P = 0.038) in faecal water significantly decreased. Total NSt concentration in faecal water was found to be significantly higher in women compared to men (P = 0.018). The genotoxicity of faecal water was neither affected by calcium supplementation nor were there gender-specific differences.Dietary calcium supplementation diversely affects BA and NSt in faeces and in faecal water but does not influence the genotoxicity of faecal water in healthy adults.
Keywords: neutral sterols; bile acids; faecal water; genotoxicity; human study

The concerted action “The process for the assessment of the scientific support for claims on foods”, PASSCLAIM, proposed criteria that could provide an international yardstick for the harmonised transparent assessment of evidence submitted to support a claim for a food or food component. The evidence would be systematically appraised against specific criteria: namely, (1) a characterisation of the food or food component to which the claimed effect is attributed; (2) human data, primarily from intervention studies that represent the target populations for the claim; (3) a dose response relationship; (4) evidence allowing for confounders such as lifestyle, consumption patterns, background diet and food matrix etc.; (5) an appropriate duration for the study; (6) a measure of compliance; (7) adequate statistical power to test the hypothesis. Validated and quality assured markers of intermediate or final outcomes could be used when ideal endpoints are not easily accessible for measurement as long as their relationship to the development of the principal outcome relevant to the claim is well characterised and substantiated. The overall coherence and totality of published and unpublished evidence should be considered in the process. Assessments for substantiation claims need expert judgement, weighting of the strength of the claim, and intelligent use of the criteria applied on an individual basis with respect both to gaps in the knowledge and to any need for new knowledge and data.
Keywords: Diet; Claims; Food and health; Markers; Health claims; Biomarkers

Although functional foods are currently regulated the same as conventional foods by the US Food and Drug Administration (FDA) there is some concern that they should not be. One concern is whether functional foods can/should carry the same type of health and nutrition labeling claims as conventional foods. For example, the type of nutrient content claim that describes a level of the nutrient such as “good or excellent source” presents a problem for functional foods since these claims relate back to a standard value for nutrients (the daily value or DV). At this time the bioactive or functional components in a functional food do not have daily values so they could not take advantage of this type of claim. Structure/function claims are also at issue since they are required to relate to the food’s attributes of taste, aroma, and nutritive value, rather than attributes of functionality (which would pertain to functional foods). There appear to be three categories of issues concerning the regulation of functional foods: safety; efficacy; and their effect on the overall diet. Since bioactive components can be synthesized or extracted and concentrated, the concern is that the amounts of these substances in functional foods might reach levels which are actually injurious to health or they may negate beneficial effects of substances in the same food. Most people/organizations consider that functional foods need to document their functionality. This means that unlike conventional foods, all functional foods, by definition, would have to apply for a health claim. Finally, the long term overarching concern is what will be the impact of a functional food-driven market on overall health. It is of interest to see how the regulatory environment for functional foods evolves in the next few years and what impact that environment has on the future of these foods.
Keywords: Functional foods; Efficacy/safety; Heath claims; USA

Vitamin D deficiency and hyperparathyroidism in relation to ethnicity: a cross-sectional survey in healthy adults by Rodrigo Moreno-Reyes; Yvon A. Carpentier; Marleen Boelaert; Khadija El Moumni; Ghislaine Dufourny; Christine Bazelmans; Alain Levêque; Christine Gervy; Serge Goldman (31-37).
The study of vitamin D status at population level gained relevance since vitamin D deficiency was recently suggested to trigger chronic disease.We aimed to describe vitamin D status, its association with bone and mineral metabolism and risk factors for deficiency in adults over 40 years in Belgium.We conducted a cross-sectional survey in a stratified random sample of 401 subjects aged between 40 and 60 years living in Brussels, and drawn from 4 different ethnic backgrounds: autochthonous Belgian, Moroccan, Turkish and Congolese. 25-Hydroxyvitamin D (25OHD), parathyroid hormone (PTH), osteocalcin, C-telopeptide and bone mineral density was measured.Three-hundred and six subjects (77%) showed 25OHD concentrations below 50 nmol/l,135 (34%) below 25 nmol/l and 18 (5%) below 12.5 nmol/l. The proportion of subjects with vitamin D deficiency was four times greater amongst those of Moroccan or Turkish descent compared with those of Congolese or Belgian descent. Moroccan subjects showed a significant higher PTH and bone marker concentrations compared to Belgian. Ethnicity, season and sex were independently associated with vitamin D deficiency in multivariate analysis.The prevalence of vitamin D deficiency is very high amongst the adult population of Brussels but immigrants are at greater risk. Given the established link between population health and adequate vitamin D status, a policy of vitamin D supplementation should be considered in these risk groups.
Keywords: vitamin D; ethnicity; bone metabolism; vitamin D deficiency; hyperparathyroidism

It is amazing to see how much the approach of the food risk analysis evolved in the recent years. For half a century and the birth of the risk assessment methodology in the food domain, only no appreciable health risk was considered acceptable by the manager. This is the vocabulary used in the case of a voluntary, deliberated human action, as the use of food additives (definition of ADI). In the case of risks not resulting from such an action, as that of the presence of contaminants, the risk assessor allocates provisional tolerable daily, weekly or monthly intake that are the basis for regulation. This vocabulary is in agreement with the objective which consists in approaching closer possible of the zero risk which is the wish of a majority of the consumers. Some years ago, the risk managers insisted to obtain from the assessors as often as possible a quantitative risk evaluation. More recently even, the managers would like to decide on the basis of a balance of risk and benefit acceptable for management purposes. Finally, they hope that general principles and tools will be available for conducting a quantitative risk-benefit analysis for foods and food ingredients. What is possible in the case of functional foods (FF)? Based on the definition of FF proposed in the programme FUFOSE, one has to distinguish between different situations in order to assess the risk: that of a micro-, that of a macro-component or that of a whole food. These situations have been clearly described in the document resulting from FOSIE. The standardized methodology relevant to assess micro-components is not well adapted to the assessment of whole food. Concepts of substantial equivalence and of history of safe use could be useful tools in this case. However, quantitative risk assessment remains a very difficult exercise. If a process for the assessment of health benefit of FF has been proposed as an outcome of the PASSCLAIM action, the quantification of this benefit needs adequate tools. An EFSA scientific colloquium on “Risk-Benefit Analysis of Foods” organized in July 2006 concluded that the risk-benefit analysis should mirror the current risk analysis paradigm and that its assessment should be performed with common scales. Disability adjusted life years (DALYs) or quality adjusted life years (QUALYs) have been proposed as some of these common scales. However, the meeting “concluded that the data available to undertake a quantitative risk-benefit assessment may be too scarce”. Because it was considered that it was premature to formulate guidelines on good risk-benefit analysis practice and it is now time to “learning by doing”, a reference to the upcoming ILSI Europe project BRAFO was done. All these aspects are discussed, in particular in relation to the specific case of FF.
Keywords: Functional foods; Safety; Risk; Benefits

Carboxymethyl-lysine, an advanced glycation end product, and decline of renal function in older community-dwelling adults by Dr. Richard D. Semba; Jeffrey C. Fink; Kai Sun; Stefania Bandinelli; Jack M. Guralnik; Luigi Ferrucci (38-44).
Advanced glycation end products (AGEs) are bioactive molecules found in greater concentrations in foods that have been processed at high temperatures. AGEs have been associated with impaired renal function in diabetes and in uremia. The relationship between AGEs and renal function in community-dwelling adults has not been well characterized.The objective was to determine whether plasma AGEs are independently associated with chronic kidney disease (CKD) and predictive of renal function in older adults.The relationship between plasma carboxymethyl-lysine (CML), an AGE, and CKD (≥ stage 3 of National Kidney Foundation classification; estimated glomerular filtration rate [eGFR] <60 ml/min/1.73 m2) and eGFR at 3- and 6-years follow-up was examined in a population-based study of aging, the InCHIANTI study, in Tuscany, Italy.Of 1,008 adults, aged ≥65 years, 153 (15.2%) had CKD at enrollment. Mean (standard deviation [S.D.]) plasma CML was 365 (110) ng/ml. Plasma CML was associated with CKD (odds ratio [O.R.] expressed per 1 S.D., 1.53, 95% confidence interval [C.I.] 1.27–1.84, P < 0.0001) in a multivariate logistic regression model, adjusting for potential confounders. Plasma CML was associated with eGFR (β = −2.77, standard error [S.E.] = 0.51, P < 0.0001) at baseline, 3-year (β = −2.54, S.E. = 0.61, P < 0.0001) and 6-year follow-up visits (β = −1.21, S.E. = 0.70, P = 0.08) in multivariate linear regression models, adjusting for potential confounders. The associations between plasma CML and prevalent CKD, eGFR, and eGFR at 3- and 6-year follow-up were significant and nearly unchanged after exclusion of adults with diabetes.Plasma CML is independently associated with CKD and is an independent predictor of decline in renal function in older community-dwelling adults.
Keywords: advanced glycation end products; aging; carboxymethyl-lysine; chronic kidney disease; renal function

Nutrition communication by means of nutrition and health claims and otherwise, holds the potential to contribute to public health by stimulating informed healthier food choices and enhanced health-focussed competition in the market place, provided that the health messages are trustworthy (i.e. scientifically substantiated) and correctly used and interpreted by the consumer. Not surprisingly, these two considerations constitute the cornerstone of the new EU legislation on nutrition and health claims, in which evidence for consumer understanding of nutrition and health claims is a new requirement.To review some of the key issues in consumer understanding of nutritional communication as a basis for reflection on the consumer understanding element of the new EU legislation on nutrition and health claims.There is a need for more methodologically advanced research in consumer understanding of nutrition and health claims as a basis for truly assessing the real-life use of such information and its actual effect on consumer food choices. Such approaches are pertinent in light of the evaluation and approval process of (new) nutrition and health claims as required under the new EU legislation on nutrition and health claims.
Keywords: Consumer understanding; Nutrition and health claims; EU legislation; Nutrition communication

Short-term effect of bedtime consumption of fermented milk supplemented with calcium, inulin-type fructans and caseinphosphopeptides on bone metabolism in healthy, postmenopausal women by Berit Adolphi; Katharina E. Scholz-Ahrens; Michael de Vrese; Yahya Açil; Christiane Laue; Jürgen Schrezenmeir (45-53).
Milk products are good sources of calcium and their consumption may reduce bone resorption and thus contribute to prevent bone loss.We tested the hypothesis that bedtime consumption of fermented milk supplemented with calcium inhibits the nocturnally enhanced bone resorption more markedly than fermented milk alone, and postulated that this effect was most pronounced when calcium absorption enhancers were added.In a controlled, parallel, double-blind intervention study over 2 weeks we investigated the short-term effects of two fermented milks supplemented with calcium from milk minerals (f-milk + Ca, n = 28) or calcium from milk minerals, inulin-type fructans and caseinphosphopeptides (f-milk + Ca + ITF + CPP; n = 29) on calcium and bone metabolism in healthy, postmenopausal women, and compared them with the effect of a fermented control milk without supplements (f-milk, n = 28). At bedtime 175 ml/d of either test milk was consumed. Fasting blood samples and 48 h-urine were collected at baseline and at the end of the intervention. Urine was divided into a pooled daytime and nighttime fraction. Multifactorial ANOVA was performed.Fermented milk independent of a supplement (n = 85) reduced the nocturnal excretion of deoxypyridinoline, a marker of bone resorption, from 11.73 ± 0.54 before to 9.57 ± 0.54 µmol/mol creatinine at the end of the intervention (P = 0.005). No effect was seen in the daytime fraction. Differences between the three milks (n = 28 resp. 29) were not significant. Fermented milk reduced bone alkaline phosphatase, a marker of bone formation, from 25.03 ± 2.08 to 18.96 ± 2.08 U/l, with no difference between these groups either. Fermented milk increased the nocturnal but not daytime urinary excretion of calcium and phosphorus. The effects on calcium and phosphorus excretion were mainly due to the group supplemented with Ca + ITF + CPP.Bedtime consumption of fermented milk reduced the nocturnal bone resorption by decelerating its turnover. Supplemented calcium from milk mineral had no additional effect unless the absorption enhancers ITF + CPP were added. A stimulated intestinal calcium absorption may be assumed, since urinary calcium excretion increased at a constant bone resorption.
Keywords: fermented milk; inulin-type fructans; caseinphosphopeptides; calcium supplementation; bone metabolism

Nutrition trends in Europe are driven by taste, health and convenience. The possibilities of research using new technologies and tools such as nutrigenomics, imaging techniques, nanotechnology, bioinformatics, cognitive sciences, innovative processes are very promising to support these nutrition trends and in particular their health aspects. This is supported by European Union research. The opportunities offered in the 7th Framework Programme (FP7), among other innovations, will contribute to the general aim of improving nutrition policy as well as improving products from the food industry in accordance with the Lisbon strategy to create employment and improve the quality of life of the European citizens.
Keywords: Functional foods; Framework Programmes; European Commission

Neuroprotective effect of cocoa flavonids on in vitro oxidative stress by Emma Ramiro-Puig; Gemma Casadesús; Hyoung-gon Lee; Xiongwei Zhu; Andrew McShea; George Perry; Francisco J. Pérez-Cano; Mark A. Smith; Margarida Castell (54-61).
Cocoa is a rich source of flavonoids that, among other functions, can act as antioxidants. In living systems, the production of reactive oxygen species (ROS) activate an array of intracellular cascades, including mitogen-activated protein kinases (MAPK), that are closely associated with cell death or survival pathways.To ascertain the role of a cocoa extract and its main flavonoid, (-)-epicatechin, in an in vitro model of oxidative stress induced in a neuronal cell line.We analyzed ROS production by fluorometry (dichlorofluorescein assay), and activation of MAPK pathways including extracellular signal-regulated kinases 1/2 (ERK 1/2), c-Jun N-terminal kinase (JNK), and p-38, by Western blot analysis.Cells incubated with cocoa extract or (-)-epicatechin, reduced ROS production in a dose-dependent manner, reaching 35% inhibition. pJNK and p38, involved in apoptosis, were down-modulated by cocoa extract and (-)-epicatechin with p38 inhibition reaching up to 70%.Our results show that cocoa extract and (-)-epicatechin may exert a neuroprotective action by reducing ROS production and modulating MAPK activation.
Keywords: cocoa; c-JNK; MAPK; neuronal damage; oxidative stress

Neuroprotective effect of cocoa flavonoids on in vitro oxidative stress by Emma Ramiro-Puig; Gemma Casadesús; Hyoung-gon Lee; Xiongwei Zhu; Andrew McShea; George Perry; Francisco J. Pérez-Cano; Mark A. Smith; Margarida Castell (61-61).

Evidence that a maternal “junk food” diet during pregnancy and lactation can reduce muscle force in offspring by Stéphanie A. Bayol; Raymond Macharia; Samantha J. Farrington; Bigboy H. Simbi; Neil C. Stickland (62-65).
Obesity is a multi-factorial condition generally attributed to an unbalanced diet and lack of exercise. Recent evidence suggests that maternal malnutrition during pregnancy and lactation can also contribute to the development of obesity in offspring. We have developed an animal model in rats to examine the effects of maternal overeating on a westernised “junk food” diet using palatable processed foods rich in fat, sugar and salt designed for human consumption. Using this model, we have shown that such a maternal diet can promote overeating and a greater preference for junk food in offspring at the end of adolescence. The maternal junk food diet also promoted adiposity and muscle atrophy at weaning. Impaired muscle development may permanently affect the function of this tissue including its ability to generate force.The aim of this study is to determine whether a maternal junk food diet can impair muscle force generation in offspring.Twitch and tetanic tensions were measured in offspring fed either chow alone (C) or with a junk food diet (J) during gestation, lactation and/or post-weaning up to the end of adolescence such that three groups of offspring were used, namely the CCC, JJC and JJJ groups.We show that adult offspring from mothers fed the junk food diet in pregnancy and lactation display reduced muscle force (both specific twitch and tetanic tensions) regardless of the post-weaning diet compared with offspring from mothers fed a balanced diet.Maternal malnutrition can influence muscle force production in offspring which may affect an individual’s ability to exercise and thereby combat obesity.
Keywords: maternal diet; junk food; skeletal muscle force; obesity; exercise