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Ultra-processed foods in human nutrition and health

Belgian Nutrition Society Eight annual meeting, May 4th, 2018

Proceedings Recently, food classification and coding systems based on the degree of processing have been introduced to redefine food-based dietary guidelines hereby taking into account the potential role of processed foods on human health. Ultra-processed foods have received special attention in view of the higher levels of salt, added sugar, fat and additives to influence the palatability of products in this food category. In this edition of the BNS annual congress, we bring a balanced view on the role of (ultra)processed foods in human nutrition and health. We will pay attention to the classification of foods based on degree of processing, and give an overall state of the art regarding the impact of processed foods on human nutrition and health. Next, we will highlight the role of processing in food science and nutrition, and we discuss the contribution of nutrient intake from processed foods, in relation to their potential role towards human health. There is also a more global view of food processing, taking into account the international trade and globalization of food and dietary patterns in both developed and developing countries. This 2018 event is supported by

The Belgian Nutrition Society has strict procedures to ensure scientific independence and integrity as described in the rules for sponsorship and applications for membership i.e. General Assembly and Board.

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Programme Welcome session 8.30 – 9.10: Registration and welcome coffee 9.10 – 9.20: Introduction by BNS - N. Delzenne (UCL- President BNS) 9.20 – 9.30: Presentation of the scientific program – J. Van Camp (UGent– Chair of the Organizing Committee)

Session 1: (Chair Nathalie Delzenne) 9:30-10:00 Keynote 1: Ultra-processed foods in human health: a critical appraisal Prof. M. Gibney (UCD Institute of Food and Health, University College Dublin, Dublin, Ireland) selected oral presentations 10:00-10:15 Lise Deroover: Wheat bran with reduced particle size does not affect gut health and systemic health in normal weight and obese subjects 10:15-10:30 Moyersoen Isabelle: Do current fortification and supplementation programs assure adequate intake of fat soluble vitamins in Belgium? Session 2: (Chair Kristin Verbeke) 10:30 – 11:00 Keynote 2: Role of food processing in food science and nutrition Prof. Frank Devlieghere (Faculty of Bio-Science Engineering, Ghent University, Belgium) selected oral presentations 11:00-11:15 Wim Geeraerts: Bacterial species diversity on vegetarian, vegan, and insect alternatives for meat in the Belgian retail 11:15-11:30 Henry Ambayo: Development of an ontology for standardizing nutritional epidemiological study metadata and improving interoperability 11:30-13:00 Lunch and poster presentations

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Session 3: (Chair Stefaan De Henauw) 13:00-13:30 Keynote 3: Ultra-processed foods, diet quality and health: summary of the evidence

Prof. Jean-Claude Moubarac (Faculté de médecine - Département de nutrition, Université de Montreal, Canada) selected oral presentations 13:30-13:45 Lisa Miclotte: Dietary Emulsifiers and the microbial gut community: Effects on SCFA-production and community structure 13:45-14:00 Vandenbroele Jolien: Curbing portion size effects of processed foods at the point of purchase 14:00-14:45 Coffee break Session 4: (Chair: John Van Camp) 14:45-15:15 Keynote 4: Ultra-processed foods and the transformation of food systems: A global policy perspective Dr. S. Vandevijvere (WIV-ISP & INFORMAS, International Network for Food and Obesity/NCDs Research, Monitoring and Action Support) Selected oral presentations 15:15-15:30 Marta Olivares: Arabinoxylan-oligosaccharides prevent obesity induced in mice by the addition of gluten in the Western diet 15:30-15:45 Mulusew J. Gerbaba: The impact of food insecurity on mental health, growth trajectories and human capital among adolescents (generation Y) of Ethiopia Closing session 15.45 – 16.00: Presentation of the BNS prizes for best communication (oral and poster) - (Nathalie Delzenne UCL- President BNS) 16.00 – 16.15: Conclusions of the symposium – John Van Camp (UGent Belgium – Chair of the Organizing Committee)

Updates and announcements will feature on our website www.belgiannutritionsociety.be

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PRACTICAL INFORMATION Venue: Royal library of Belgium (near the central station in Brussels) Koninklijke Bibliotheek van België - Royal Sky Room / Keizerslaan 2, 1000 Brussel

Plan: http://www.kbr.be/images/informations/divers/plan_nl_gd.jpg

Bibliothèque royale de Belgique - Royal Sky Room / boulevard de l'Empereur 2, 1000 Bruxelles

Plan: http://www.kbr.be/images/informations/divers/plan_fr_gr.jpg

How to get there: Detailed information: http://www.kbr.be/informations/informations_nl.html Catering: Food and drinks will be provided for free for participants during the breaks. Language: English. No translations are provided.

Registration

All participants need to register electronically. The congress is free for BNS members. There will

be an opportunity to register as BNS member on-site (please consult membership procedures and

application form on our website (www.belgiannutritionsociety.be/ go2/Membership/) for more info.

Membership fees are 35€ and 15 € for students and PhD students.

Speakers and posters

The program of the symposium consists of different sessions. Each session has a keynote

speaker who will give a “state of the art” lecture. Each keynote lecture of all sessions is followed by

‘local’ speakers. For this part of each session, young Belgian scientists will be selected on the basis

of the submitted abstracts to orally present their research work. These parts of the sessions aim to

provide postgraduate students, PhD students or scientists at all stages of their research, the

opportunity to present their work (via oral or ‘live’ poster presentations), independently of the topic of

the symposium. Deadline of abstracts is March 30th, 2018.

Organising Committee

John Van Camp (Chair, UGent)

Stefaan De Henauw (UGent)

Kristin Verbeke (KULeuven)

Nathalie Delzenne (UCLouvain)

Carl Lachat (UGent)

Patrick Kolsteren (UGent)

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Oral presentations

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Ultra-processed foods in human health: a critical appraisal

by

Michael J Gibney

There are two routes to the improvement of the national diet, each of which plays a role. The first route is to change food choice, directing people away from one type of food to an alternative i.e. substitute white meat for red meat or fish for meat in general; choose boiled potatoes over fried potatoes or chips; eat fruit instead of snacks. The second route is to allow subjects to continue enjoying their traditional food habit but change the nature of the food choice: switch to low fat milk, to MUFA spreads, to wholegrain breads, to higher fibre cereals, to diet soft drinks. As regards the latter, considerable governmental and international commitments to the process of ‘reformulation’ are underway. The NOVA approach identifies four categories of foods and lists one to be avoided, ultra-processed foods (UPF) and one to be reduced, processed foods. Emphasis is given to the promotion of fresh and unprocessed foods.

A major difficulty in understanding the nature of UPF is its extremely vague definition. Part of this definition relates to salt, sugar and fat with no guidance as to cut off levels for consideration, the base unit of expression (per 100g, per 1000kJ or per serving size) or the population impact of a given food (% total daily intake) to the consumption of the three nutrients. The second element of the definition of UPF relates to the role that food additives play in food. In this context, the specific proposed impact of such additives is to ‘mask undesirable qualities’ and to ‘imitate sensorial qualities’ of unprocessed foods. The former would be a challenge not only to consumer acceptability but also to food safety legislation. The latter is difficult to interpret if an unprocessed food is deemed to imitate some unprocessed version when the latter is not obvious: ice cream, soft drinks, margarine, breakfast cereals.

By definition, UPF excludes fresh fruit and vegetables, fresh meat, fish and poultry, milk, eggs, cheese, butter, wine, fresh bread and other non-processed foods (in reality cheese, butter, wine, bread etc. are processed since ancient times). Thus, by definition, UPF has a low nutrient density and by virtue of a pre-defined high level of sugar and fat, has a high energy density. Not surprisingly, given it’s a priori but poorly defined nature, UPF will always be associated with lower fibre and micronutrient intakes and higher intakes of fat, saturated fat and sugar. Any public health nutrition campaign advocating the absolute avoidance of a poorly defined food category, such as UPF, is unlikely to be successful. The scale of social change required to effect a difference in the national diet would be too great. In contrast, the use of nudge theory to move populations toward a healthier diet is far more likely to succeed. Within that strategy, major reformulation of processed food is more likely to effect change than the abandonment of the poorly defined concept of UPF.

Institute of Food and Health, University College Dublin, Ireland

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Wheat bran with reduced particle size does not affect gut health and systemic health in normal weight and

obese subjects

by

Lise Deroover1, Joran Verspreet2,3, Greet Vandermeulen1, Anja Luypaerts1, Christophe Courtin2,3 and Kristin Verbeke1,3

Introduction and objectives Wheat bran (WB) consumption has been associated with improved gastrointestinal (GI) health and a reduced risk for metabolic disorders. Reducing the particle size of WB decreases its bulking effect but increases its fermentability. We investigated whether WB with reduced particle size (WB RPS) still affects gut and systemic health. In addition to normal weight (NW) subjects, obese (OB) subjects were included because of their presumed suboptimal health.

Materials & Methods During 1 month, 36 NW (BMI 22 ± 1.23 kg/m²) and 14 OB subjects (BMI 33 ± 2.13 kg/m²) daily consumed 20 g WB RPS or placebo (PL). Before and after the intervention, faecal wet weight and % dry matter, fat excretion, stool consistency (Bristol Stool Score) and faecal SCFA concentrations were quantified. Total GI transit was measured using 3H-PEG and expressed as % 3H recovered. Intestinal permeability was assessed using a 24-h 51Cr-EDTA urinary excretion test. Total cholesterol (TC) and triglyceride (TG) levels and C-reactive protein (CRP) levels were measured in blood as parameter of lipid metabolism and systemic inflammation, respectively. Baseline parameters were compared between NW and OB subjects using a Mann-Whitney U test. A two-way ANCOVA was performed to compare the effect of the WB and PL intervention between both BMI groups. A significance level p<0.05 was used.

Results Baseline faecal wet weight and % dry matter, fat excretion, GI transit and stool consistency were not different between NW and OB subjects. Similarly and despite literature data, SCFA concentrations were comparable in NW and OB (131.2 ± 64.7 mmol/L vs. 129.3 ± 64.9 mmol/L, respectively). There was no effect of treatment on faecal wet weight and % dry matter, GI transit, stool consistency and SCFA concentrations, irrespective of BMI. However, we found a significant interaction between treatment and BMI on fat excretion, due to a more pronounced decrease in fat excretion after WB consumption in OB than in NW subjects (p=0.044).

Unexpectedly, small intestinal and colon permeability were not higher in OB (0.68 ± 0.23 % and 0.84 ± 0.31%, respectively) than in NW (0.66 ± 0.36 % and 0.97 ± 0.31 %, respectively). In none of the groups, WB consumption affected permeability. Plasma TC and TG levels were not different between NW and OB subjects at baseline (153.3 ± 31.7 mg/dL and 74.3 ± 26.6 mg/dL for NW and 171.5 ± 35.6 mg/dL and 86.5 ± 32.6 mg/dL for OB) and were not influenced by the intervention. Significantly higher CRP levels in OB compared to NW subjects (3.70 ± 3.21 mg/L vs 1.46 ± 1.89 mg/L; p=0.003) confirmed their low-grade inflammation, yet were not decreased by WB treatment.

Conclusion WB RPS did not induce health benefits that have previously been associated with consumption of coarse WB. OB subjects were not more susceptible to WB-induced effects than NW subjects, possibly because of insufficient overweight.

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Keywords: wheat bran, wheat bran modification, dietary fibre, colonic fermentation, colonic health, systemic health 1 Translational Research in Gastrointestinal Disorders, KU Leuven, Leuven, Belgium 2 Centre for Food and Microbial Technology, KU Leuven, Leuven, Belgium 3 Leuven Food Science and Nutrition Research Centre, KU Leuven, Leuven, Belgium e-mail: lise.deroover@kuleuven.be

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Do current fortification and supplementation programs assure adequate intake of fat soluble vitamins in

Belgium? by

Moyersoen Isabelle1,3, Karin De Ridder1, Brecht Devleesschauwer1, Stefanie Vandevijvere1, Koenraad Cuypers1, Jean Tafforeau1, Arnold Dekkers2, Margot Van steenland3, Bruno De

Meulenaer3, Herman Van Oyen3,4, Carl Lachat3, John Van Camp3

Aims of the study: To evaluate the risk for inadequate and excessive intakes of fat soluble vitamins from fortified food and supplement consumption.

Methods: The study was conducted in the general Belgian population (3-64 year) and four at risk groups, i.e. infants, toddlers, pregnant and lactating women. Consumption data of the general population was retrieved from the Belgian food consumption survey (BFCS2014) using 2 24h recalls and a food frequency questionnaire (FFQ). Consumption data of the vulnerable groups were collected with a FFQ comprising the top 90% food groups contributing to the intake of vitamins A, D, E and K and fortified foods retrieved from a market research. Data were analysed in Spade and SAS respectively.

Main findings: The risk for inadequate intake of vitamin E and K was low. Inadequate intakes of vitamin A were found in all study-populations except infants (0-11 months). The prevalence increased with age and was highest in adolescents (girls, 26%; boys, 34-37%) and lactating women (35%). During infancy, non-users of vitamin D prophylaxis (22%) were at risk for inadequate vitamin D intake, while consumption of highly dosed supplements posed a risk for excessive intakes (6-26%). In all other study populations median vitamin D intake was less than one third of the adequate intake (AI=15μg/day) which predispose subjects with minor sun-exposure for vitamin D deficiency. Contribution of fortified foods to vitamin A, D, E, and K intake was minor, except in toddlers.

Conclusions: Belgian fortification and supplementation practices need to be revised to enhance vitamins A and D intake at the population level.

Keywords: Belgian population, vulnerable groups, fat soluble vitamins, dietary intake

assessment, fortified foods, supplements

1 Department of Public Health and Surveillance, Scientific Institute of Public Health (WIV-ISP), Juliette Wytsmanstraat 14, 1050 Brussels, Belgium; isabelle.moyersoen@wiv-isp.be ; Karin.DeRidder@wiv-isp.be; Stefanie.Vandevijvere@wiv-isp.be; Brecht.devleesschauwer@wiv-isp.be; Koenraad.Cuypers@wiv-isp.be;

Jean.Tafforeau@wiv-isp.be; herman.vanoyen@wiv-isp.be; 2Department for Statistics, Informatics and Modelling, National Institute for Public Health and the Environment (RIVM), P.O. BOX 1, 3720 BA Bilthoven, The Netherlands; arnold.dekkers@rivm.nl; 3 Department of Food Safety and Food Quality, Ghent University, Coupure Links 653, 9000 Ghent, Belgium; Margot.vansteenland@UGent.be; bruno.demeulenaer@UGent.be; Carl.lachat@Ugent.be; john.vancamp@ugent.be; 4Department of Public Health, Ghent University, De Pintelaan 185, 9000 Gent, Belgium

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Role of food processing in food science and nutrition

by

Frank Devlieghere, Bruno De Meulenaer, André Huyghebaert, Patrick Kolsteren, Carl Lachat, Katleen Raes, Imca Sampers, Iris Tavernier, Mieke Uyttendaele, John Van Camp

Recently, the Flemish food triangle considered ultra-processed foods, as described by the NOVA classification, to be avoided as much as possible. Because the NOVA definition of (ultra) processed foods is mainly based on the complexity of the product and not so much on the degree of processing, the consumer might as a result of this recommendation avoid processed foods, as defined from a technological point of view. In this presentation the objectives of food processing are discussed and examples are given on how food processing is influencing negatively as well as positively the nutritional properties of foods. Reflections are made on the possible consequences of the recommendation to avoid ultra-processed foods and alternative actions are given to improve the nutritional quality of food products.

Department of Food Technology, Safety and Health. Faculty of Bioscience Engineering, Ghent University Belgium

E-Mail: frank.Devlieghere@UGent.be

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Bacterial species diversity on vegetarian, vegan, and insect alternatives for meat in the Belgian retail

by

Wim Geeraerts, Luc De Vuyst, Fréderic Leroy1

Recently, vegetarian, vegan and insect products are becoming popular alternatives for meat products. These products are processed to varies degrees. Like modified-atmosphere-packaged cooked meat products, they are potentially at risk of bacterially induced spoilage, although information on their microbial species diversity is limited. In this study, a market study of several vegetarian/vegan products as well as formulations of the edible insect species Galleria mellonella (greater wax moth) and Gryllodes sigillatus (tropical house cricket) was performed

Eighteen samples from processed vegetarian/vegan products and six samples from insect products were analysed at expiration date. Four additional vegetarian/vegan samples were analysed after temperature abuse. Bacterial analysis was done using a combination of pour and spread plating on ten different agar media to cover different microbial groups. Colonies were picked up (5 to 30 % from the highest dilutions) and subjected to DNA extraction and (GTG)5-PCR genomic fingerprinting, followed by numerical cluster analysis. Identification of representative isolates was done by sequencing of the 16S rRNA, tuf, and pheS genes.

In general, the bacterial counts on the vegetarian/vegan samples examined were relatively low, ranging from < 2.0 to8.3 log (cfu g-1). On eleven samples, the bacterial loads were below 2.0 log (cfu g-1) on all agar media used. The bacterial counts on the insect samples ranged from < 2.0 to 8.1 log (cfu g-1) on the different agar media. The most commonly identified isolates from the vegetarian/vegan samples (327 isolates) were Enterococcus faecium (113 isolates) and Lactobacillus sakei (151 isolates). Enterococcus faecium (146 isolates) was the most common species on the insect products (219 isolates). Clear differences were found between the different types of vegetarian/vegan products and insect species, indicating that in-depth knowledge about the bacterial communities on these upcoming products is necessary in view of their optimal processing and thus market stability.

Keywords: vegetarian/vegan products, edible insects, bacterial spoilage, lactic acid bacteria

1 Research Group of Industrial Microbiology and Food Biotechnology (IMDO), Faculty of Sciences and Bioengineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussel, Belgium. Wim.geeraerts@vub.be

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Development of an ontology for standardizing nutritional epidemiological study metadata and

improving interoperability

by

Henry Ambayo1, Carl Lachat1, Chen Yang*1

Background: Integrating data can increase power of nutrition research and enable pooled analysis, and linkages with other disciplines related to genome and phenotype. Existing data repositories however have heterogeneous approaches to described data, which impairs machine-readability of data. An ontology, i.e. the description of a knowledge domain in a machine-readable format, can overcome these limits but is currently not available for nutrition research.

Aim: The objective of this study was to develop an ontology according to Minimal Data

Requirements (MDR) for description of metadata for nutrition studies.

Methods: The MDR is a set of minimal requirements for study metadata based on collation of relevant, specific nutrition epidemiological concepts. Concepts with multiple choice options were selected because the range of search responses are defined as opposed to filling in blanks. The Ontology for Nutritional Epidemiology- MDR(ONE-MDR) was constructed using Ontology Web Language (OWL Working Group, 2012). A case study application was conducted to demonstrate the practical application of the ONE-MDR ontology using a web tool.

Main findings: The ONE-MDR classes were categorized into general descriptors of the investigation, study and assay descriptors. Overall, 44 classes, 21 object properties relevant for search purposes and 16 datatype properties were created.

Principle conclusion: The ONE-MDR includes essential terms to describe study metadata in a machine-readable format, thus facilitates easy search and findability of study datasets.

Keywords: ontology, minimal data requirements, nutrition epidemiology, metadata, study

1 Department of Food Technology, Safety and Health, Ghent University, Ghent, Belgium Email: *Chen.Yang@UGent.be

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Ultra-processed foods, diet quality and health: summary of the evidence

by Jean-Claude Moubarac

In this presentation I will argue that although many types of food processing have been and

are essential for human life, ultra-processed foods are intrinsically harmful for human health, human culture, and the planet sustainability. I will present the NOVA food classification system, within each ultra-processed foods are defined, descriptive statistics showing worldwide increases in the sales and consumption of ultra-processed foods and a summary of evidence that supports the thesis that these products are intrinsically detrimental. I will conclude discussing the policies we need and the main obstacles we face to transform current food trends and systems into healthy and sustainable ones.

Département de nutrition, Faculté de médecine

Université de Montréal

Email: jc.moubarac@umontreal.ca

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Dietary emulsifiers and the microbial gut community: Effects on SCFA-production and community structure

by

Lisa Miclotte1, I. Van Bogaert2, T. Van de Wiele1

Background and aims: Dietary additives are widely used in processed food products and have become a target for research in terms of their impact on the gut microbiome. Recent studies with dietary emulsifiers in both in vitro and murine models have indicated a strong impact on both the gut microbiome and host health, possibly related to chronic gut inflammation and other western ailments. Some observed effects are an altered microbiome composition, an impaired gut barrier, increased expression of gut inflammatory compounds, increased food intake and weight gain.

This study aims at delving deeper into the effects of 5 dietary emulsifiers on the composition

and the activity of the gut bacteria. A comparison is made between 2 mainstream chemical ones (Carboxymethylcellulose and Tween80) and 3 natural or biotechnological examples (Soy lecithin, Sophorolipids and Rhamnolipids).

Set-up: The short terms effects of the 5 emulsifiers on the gut microbiota of several individuals were tested in a series of 48h batch-incubations, incorporating 5 emulsifier concentrations (max 0.5%). The endpoints measured were the bacterial activity (SCFA-production), the intact cell population (SGPI-staining and Flow cytometry) and community composition (MiSeq sequencing).

Results: The 5 emulsifiers show differential effects on both SCFA-production and cell survival,

indicating the novel Sophorolipids and Rhamnolipids as more harmful. Importantly, both seem to decrease the butyrate fraction, in favour of propionate. Carboxymethylcellulose and Tween80 show limited effects on SCFA-production and cell counts. Soy Lecithin’s impact is more noticeable in the cell count data than on SCFA-level.

Keywords: Gut microbiota, Dietary Emulsifiers, Batch Experiment, Interindividual variablility, Microbiome composition, Microbial activity. 1Cmet (Center for Microbial Ecology and Technology), Department of biochemical and microbial technology, Faculty of Bioscience Engineering, Ghent University, Belgium Lisa.Miclotte@UGent.be Tom.Vandewiele@UGent.be

2Inbio (Centre of Expertise for Industrial Biotechnology and Biocatalysis (INBIO), Department of biochemical and microbial technology, Faculty of Bioscience Engineering, Ghent University, Belgium

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Curbing portion size effects of processed foods at the point of purchase

by

Vandenbroele Jolien1, Hendrik Slabbinck1, Anneleen Van Kerckhove1, Iris Vermeir1

In recent decades, portion sizes have increased tremendously, prompting consumers to eat

more than recommended. Besides, this trend is aligned with higher CO2-emission rates because of increased food production volume. This is especially worrying for less sustainable products, such as processed red meat that generates a 150% more CO2-emission compared to poultry meat. This study takes a behavioural perspective to determine whether adding smaller portion sizes to a retailer’s assortment unobtrusively encourages consumers to buy smaller portions of processed meat. Therefore, a field experiment with meat sausage as focal product, was conducted over the course of a month in a branch of a large European retailer, generating receipts of each individual shopping trip. The product sales data revealed slightly more sales of the two smaller portions in terms of units (52%), versus the default, larger portion (48%), resulting in a pertinent reduction in total volumes sold in kg. Furthermore, a two-way ANOVA comparing sales at the individual level before and during the intervention for the experimental store versus eight control stores showed a reduction in the volume of meat sold during the intervention period in the experimental store. Moreover, the results of a one-way ANOVA indicated that smaller portion buyers do not compensate by buying more other products within the same product category. The finding that adding smaller portions to a default choice architecture can nudge consumers towards buying smaller sized items, has important implications for retailers and public policy makers involved in promoting healthy and sustainable consumer behavior.

TABLE 1. Overview of the data by research question

Keywords: Portion size; Choice architecture; Grocery shopping; Sustainable consumer behaviour

1 Ghent University, Faculty of Economics and Business Administration, Department of Marketing, Tweekerkenstraat 2, 9000 Ghent, Belgium. E-Mail: Jolien.vandenbroele@ugent.be

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Ultra-processed foods and the transformation of food systems: A global policy perspective

by

Dr Stefanie Vandevijvere

Recently it has been proposed that food processing, more specifically the type, intensity and purpose of food processing, may be linked to human health. Apart from Western Europe, North America and Australasia, volume sales per capita of ultra-processed food products increased over the last 15 years, particularly in South and South East Asia. Some countries, such as Brazil, Uruguay and France, now recommend limits on the consumption of ultra-processed foods within their food-based dietary guidelines. In addition, the Pan-American Health Organization (PAHO) developed a nutrient profile model for processed and ultra-processed foods and encourages countries to apply this model when developing specific nutrition policies. Globally, although some progress has been made on the implementation of healthy food policies, particularly in regards to taxes on sugar-sweetened beverages and front-of-pack labelling, progress is still slow and inadequate. Accountability systems need to be strengthened to accelerate the implementation of healthy food policies and reduce global obesity. The International Network for Food and Obesity/non-communicable diseases Research Monitoring and Action Support (INFORMAS) has developed tools and processes to increase accountability of the major actors, governments and the private sector, to implement widely recommended healthy food policies, in particular the Healthy Food Environment Policy Index (Food-EPI) and the Business Impact Assessment on Obesity and Population Level Nutrition (BIA-Obesity). These upstream monitoring tools can catalyze the formulation and implementation of SMART commitments by governments and industry to support the United Nations Decade of Action on Nutrition 2016-2025.

WIV-ISP & INFORMAS, International Network for Food and Obesity/NCDs Research, Monitoring and Action Support

E-mail: Stefanie.Vandevijvere@wiv-isp.be

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Arabinoxylan-oligosaccharides prevent obesity induced in mice by the addition of gluten in the Western diet

by

Marta Olivares1, Audrey M. Neyrinck1, Julie Rodriguez1, Laure B. Bindels1,

Nathalie M. Delzenne1

Gluten-containing foods are common in the Western diet (WD) due to the high consumption of cereals and to the addition of gluten for technological purposes in processed products. The impact of gluten in the context of obesity remains poorly studied, but the limited available references describe an obesogenic effect. We aimed to investigate the effect of gluten addition in a WD and to unravel the potential effect of prebiotics in this model.

45 male C57BL/6J mice were randomized in: 1) Control diet; 2) WD; 3) WD + 5% gluten

(WD+G); 4) WD+G + 5% arabinoxylans (WD+G+AXOs); 5) WD+G + 5% fructooligosaccharides (WD+G+FOS).

WD increased the body weight and the white adipose tissue. The addition of gluten aggravated

both effects. Among the prebiotics, only AXOs significantly lessened adiposity and decreased the size of adipocytes compared to the WD+G group. WD with gluten increased the gut permeability (assessed by the translocation of FITC-dextran), reduced the expression of Occludin, and increase Interferon-γ in the ileum. AXOs trended to reduce the expression of Interferon-γ. The effects of gluten were not related to changes in any of the bacterial members analysed until now (sequencing in progress), whereas AXOs effects must be analysed taking into account the increase in Bifidobacterium spp.

Our study describes the contribution of gluten in the development of obesity and the protective effect of AXOS in an animal model. Further analyses are ongoing to unravel the mechanisms behind the obesogenic effects displayed by gluten, as well as and the protective one of AXOs. Keywords: Gluten, Western diet, prebiotics, arabinoxylan, oligosaccharides, obesity

1Metabolism and Nutrition Research Group, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium E-mail: marta.olivares@uclouvain.be

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The impact of food insecurity on mental health, growth trajectories and human capital among adolescents

(generation Y) of Ethiopia

by

Mulusew J. Gerbaba1,2, David Lindstrom2, Nathalie De Cook3, Tefera Belachew1, Carl Lachat2, Craig Hadley4, Patrick Kolsteren2

Study aim: We examined the impact of food insecurity on mental health, growth trajectories and cognitive skills of adolescents in Ethiopia.

Methods: We used the Jimma Longitudinal Family Survey of Youth (JLFSY) (n=2084) and Young Lives (YL, UK, (n=2000). Appropriate statistical analyses were computed and interpreted accordingly.

Results: The prevalence of CMDs was 30.8 % (95% CI: 28.6, 33.2), poor self-rated health status was relatively unstable (18.9%-37.3 %), the magnitude of food insecurity also ranges from 20.4%-48.4%. In this study, food insecurity was independently associated with CMDs (β= 0.323, P<0.05), the relationship was mainly direct (92.8%) and only 7.2% was partially mediated through physical morbidity. Early exposure to food insecurity is also strongly associated with mental health(P<0.05), poor self-rated health (P<0.001), BMI growth trajectories(P<0.05) and adolescent girls (P<0.05). In addition, food insecurity and poor linear growth trajectories were linked to educational attainment, and poor cognitive skills (Math Score and PPVT test).

Conclusion: These findings reveals the need to reframe the issues of adolescents beyond existing sexual and reproductive health to other health dimensions such as poor health, mental health, nutrition and cognitive skills. It also highlights the importance of considering early life factors, urban settings, low income households, and adolescent girls. Therefore, any social, nutrition and public health interventions designed to improve the health and wellbeing of adolescent health the social determinants of health such as food insecurity into account. Further research on the other social determinants of adolescents’ health and are warranted to strengthen and support ongoing interventions in Ethiopia.

keywords: Food insecurity, adolescents, Ethiopia, mental health, growth

Affiliation: 1. Population and Family Health, Jimma University,Jimma,Ethiopia, 2. Department of Food Safety and Food Quality,Ghent University, Ghent,Belgium, 3, Department of Sociology, Brown University, Providence,Rhode Island,USA, 4. Deparment of Anthropology, Emory University, Atlanta GA, USA

Contact Address: mulusew.gerbaba@gmail.com; MulusewGerbaba.Jebena@UGent.be

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Poster presentations (alphabetically)

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Diet as moderator in the association of sedentary behavior with inflammatory biomarkers among

adolescents in the HELENA study

by

Aline Arouca1

Aim: To demonstrate that a healthy diet might attenuate the positive sedentary-inflammation relation, whereas an unhealthy diet may increase the inflammatory effect of sedentary behavior.

Methods: In 618 adolescents (13-17y) of the European HELENA study, data was available on body composition, a set of inflammation markers, and food intake determined by a self-administered computerized 24h-dietary recall for 2 days. A 9-point Mediterranean diet Score was used as dietary parameter and tested as moderator. A set of low-grade inflammatory parameters was used as outcome: several cytokines in an inflammatory ratio, C-reactive protein, 3 cell-adhesion molecules, 3 cardiovascular risk markers and 3 immune cell types. Sedentary behavior was reflected by reported total screen time. Multiple linear regression analyses tested moderation by diet in the sedentary behavior-inflammation association adjusted for age, sex, country, puberty, adiposity (sum of six skinfolds), parental education, and socio-economic status.

Results: Mediterranean diet was significant protective moderator in the effect of sedentary

behavior on alanine-transaminase enzyme (P= 0.014), and on the pro/anti-inflammatory cytokine ratio (P= 0.001), but not on other inflammatory parameters.

Conclusion: A higher adherence to the Mediterranean diet may attenuate the concentration

of some inflammatory biomarkers caused by sedentary behavior, whereas a poor diet seems to increase inflammation.

Key words: cytokines, screen time, Mediterranean diet, adolescents, HELENA study,

moderation.

1 Ghent University – Faculty of Medicine and Health Sciences - Department of Public Health - Unit Nutrition and

Food Safety E-Mail: aline.barbedoarouca@ugent.be Supervisor: Stefaan De Henauw Co-supervisor: Nathalie Michels

21

Table 1. Moderator effect of Mediterranean diet adherence on the relation between sedentary behavior and low-grade inflammation markers in 618 adolescents from the HELENA study.

Interaction term with

Mediterranean diet

Inflammation markers

(outcome)

B LLCI ULCI P

Pro/anti ratio (z-score) -0.0082 -0.0132 -0.0031 0.001

CRP (log mg/L) -3.156 x10-5 -2.805 x10-4 2.174 x10-4 0.803

WBC (log 10^/µL) 1.048 x10-5 -3.259 x10-5 5.355 x10-5 0.632

Lymphocytes (10^3/µL) -0.0024 -0.0059 0.0009 0.158

CD3 (T-cells) (%) -1.349 x10-5 -3.252 x10-5 5.54 x10-6 0.164

sVCAM-1 (log ng/mL) 1.46 x10-6 -5.224 x10-5 5.516 x10-5 0.957

sICAM-1 (log ng/mL) 7.09 x10-6 -8.310 x10-5 9.729 x10-5 0.877

sE-selectin (log ng/mL) -4.935 x10-5 -1.342 x10-4 3.558 x10-5 0.254

GGT (log U/L) -3.533 x10-5 -8.297 x10-5 1.231 x10-5 0.145

ALT (log U/L) -7.003 x10-5 -1.260 x10-4 -1.403 x10-5 0.014

Homocysteine (log µmol/L) 8.5 x10-7 -6.680 x10-5 6.851 x10-5 0.980

Moderation was tested using interaction (by Process macro for SPSS) based on including the diet*sedentary behavior predictor, next to diet, overall adiposity (sum of six skinfold thickness), age, center, socioeconomic status, maternal and paternal educational status in the prediction of inflammatory parameters. TGFβ-1, Transforming growth factor beta 1; TNF-α, Tumor necrosis factor alpha; pro/anti-inflammatory cytokines ratio ((TNF-α + IL-6)/(TGFβ-1 + IL-10)); CRP, C-reactive protein; WBC, White blood cells; CD, Cluster of differentiation; sVCAM-1, sICAM-1, sE-selectin, Soluble cell adhesion molecules; GGT, Gamma-glutamyl transferase; ALT, Alanine transaminase. B= unstandardized coefficient, LLCI and ULCI, lower and upper confidence interval respectively, β= standardized coefficient. Bold: statistical significance when P < 0.05.

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Figure 1. Moderation effect of the Mediterranean diet on the relation between inflammation-related biomarkers and sedentary behavior (A; B).

Legend:

Sedentary behavior is based on total screen time (i.e., using computers, watching TV, playing video games). In the case of a significant moderation, the sedentary behavior-inflammation relation was tested for 3 representative groups: those at the mean (average adherence), at 1 SD below the mean (lower adherence), and 1 SD above the mean (higher adherence) of the dietary index.

(A) Moderator effect of the adherence to the Mediterranean diet on alanine transaminase enzyme in sedentary behavior in all subjects.

(B) Moderator effect of the adherence to the Mediterranean diet on pro/anti-inflammatory cytokines ratio in sedentary behavior in all subjects.

23

Blinded acute gluten challenge induces extra-intestinal but not gastrointestinal symptoms in Non-Coeliac

Gluten Sensitivity by

Julie Iven1, Annelies Geeraerts 1, Tim Vanuytsel 1, Jan Tack1, Lukas Van Oudenhove1, Jessica R. Biesiekierski1

Non-coeliac gluten sensitivity (NCGS) is characterised by gastrointestinal (GI; e.g. bloating) and extra-intestinal (e.g. fatigue) symptoms, that disappear after dietary exclusion of gluten, in the absence of coeliac disease. Previous research has shown that a 3-day exposure to gluten increases depression scores in self-reported NCGS patients, however acute effects of gluten were not investigated. We tested the effect of acute single-blind administration of 16g gluten or whey protein (placebo), mixed in 250ml low-fat, unsweetened yoghurt, on psychological and GI symptoms in 20 healthy volunteers (HV) and 10 NCGS patients. GI symptoms (bloating, cramps) and extra-intestinal symptoms (fatigue, tension) were assessed using visual analogue scales every 15’ until 180’ after administration. Responses over time (compared to pre-administration) were analysed using (generalized) linear mixed models. After acute administration of gluten compared to placebo, fatigue scores increased in NCGS patients (p = 0.015, but not in HV (p = 0.744). Similarly, NCGS patients had higher tension scores after gluten compared to placebo (p = 0.043), contrary to HV (p = 0.305). NCGS patients showed more bloating (p < 0.0001) and pain (p = 0.019) compared to HV, regardless of whether gluten or placebo were ingested. These findings provide new insights in NCGS, which might be characterised by acute gluten-induced increases in extra-intestinal symptoms such as fatigue, rather than GI symptoms.

Keywords: gluten, non-coeliac gluten sensitivity, food tolerances, gastrointestinal disorders

1 Translational Research Center for Gastrointestinal Disorders, KU Leuven, Belgium Julie.iven@kuleuven.be

24

Dietary trans-fatty acid intake in relation to cancer risk: a systematic review

by

Michels Nathalie1, Van der Meulen Kristof, Vantomme Annelies, Huybrechts Inge

Purpose: A western-type diet is probably the key modifiable risk factor towards cancer. Ultra-processed foods may contain trans-fatty acids produced during partial oil hydrogenation. Harmful cardiovascular effects of trans-fatty acids are already proven, but the link with cancer risk has not been summarized yet.

Methodology: The electronic databases PubMed (n=375 articles) and Embase (n=122 articles) were used to find observational studies on the intake of trans-fatty acids as predictor of neoplasms. Eventually 24 articles were included for review.

Results: Seven cancer types have been researched in cohort and case-control studies on trans-fatty acids: breast cancer (n=6), colorectal cancer (n=9), mouth/pharynx/oesophagus cancer (n=3), non-Hodgkin lymphoma (NHL, n=4), ovarian cancer (n=5), pancreatic cancer (n=3) and prostate cancer (n=6). Univocal positive associations were found for NHL and oral/pharynx/oesophagus cancer, no significance for pancreatic cancer and contradictory results for other cancers. Concerning subtypes of trans-fatty acids: C16:1 and C18:2 but not C18:1 were related to breast cancer; C16:1 but not C18:1 or C18:2 were related to pancreatic cancer. Vulnerability factors (moderators) were genetics (prostate cancer), Caucasian ethnicity (prostate or colorectal cancer), menopause (breast cancer), high age (colon and prostate cancer) and high BMI (colon cancer).

Conclusion: This systematic review suggests increased cancer risks due to high consumptions of trans-fatty acids, especially for NHL and oral/pharynx/oesophagus cancer. Chronic inflammation and oxidative stress are possible pathways. Future studies should apply higher methodological quality (confounders and biomarkers). For targeted prevention, studies should examine which trans-fatty acid subtypes are more carcinogenic and what populations are at highest risk.

Keywords: diet, trans-fatty acid, cancer, systematic review, observational study

1 Department of Public Health, Ghent University, Belgium. E-mail: Nathalie.michels@ugent.be

25

Targeting the gut microbiota by dietary fibre and polyphenols improve the cardiometabolic dysfunction

in mice by

Audrey M. Neyrinck*1, Emilie Catry*1, Bernard Taminiau2, Georges Daube2, Chantal Dessy3 and Nathalie M. Delzenne1

Aim: Chitin-glucan (CG) is an insoluble fibre with prebiotic properties whereas pomegranate peel extracts (PPE) is rich in polyphenols prone to modulate inflammation. We tested the impact of CG and the combination of CG+PPE on gut microbiota and endothelial and inflammatory disorders in a mouse model of cardiovascular disease.

Methods: Male Apoe knock-out (KO) mice were fed a control (CT) or a high fat (HF) diet supplemented with or without 5% CG (Kitozyme, Belgium) and a combination of 5% CG and 0.5% PPE (Oxylent, Belgium) for 8 weeks.

Main findings: HF-fed KO mice presented endothelial dysfunction as attested by increasing abundance of caveolin-1 and decreasing ser1177-phosphorylated eNOS form in thoracic aorta whereas vasodilation to cumulative doses of acetylcholine was reduced in preconstricted carotid and mesenteric arteries. GC and CG+PPE reduced inflammatory markers in the liver and in the adipose tissues without changing caveolin-1 and eNOS phosphorylation. Furthermore, the combination CG+PPE improved the prostacyclin-dependent relaxation of carotid arteries. Among specific changes of gut microbiota composition, a significant increase in Lachnospiraceae family (Blautia genus) and a significant decrease in Rikenellaceae family (Alistipes genus) were observed in CG+PPE-treated mice.

Conclusion: Food supplementation with CG and PPE exerted systemic anti-inflammatory effects and restored endothelial dysfunction in carotid arteries from HF-fed KO mice. This study allows to point out gut bacteria, namely Blautia and Alistipes, that could be implicated in the management of endothelial and inflammatory dysfunctions associated with cardiovascular diseases, and unravel the role of nutrition in the modulation of those bacteria.

Keywords: endothelial dysfunction, gut microbiota, insoluble fibre, chitin-glucan,

pomegranate, polyphenolic compounds

Grant Support: This research is supported by the competitive cluster Wagralim from Wallonia (SPW-DGO6: ADIPOSTOP)

1Metabolism and Nutrition research group, Louvain Drug Research Institute, Université catholique de Louvain 2Fundamental and Applied Research for Animal and Health (FARAH), Faculty of Veterinary Medicine, University of Liège 3Pole of Pharmacology and Therapeutics, Institut de Recherche Expérimentale et Clinique, Université catholique de Louvain * contributed equally to this work audrey.neyrinck@uclouvain.be

26

Intestinal NAPE-PLD is involved in metabolic adaptation in mice model of diet-induced obesity

by

Amandine Everard 1,9, Hubert Plovier1,9, Matthias Van Hul1, Marialetizia Rastelli1 , Lucie Geurts1, Céline Druart1, Sylvie Robine2, Nathalie M. Delzenne1, Giulio G. Muccioli3, Nicolas Flamand4, Willem

M. de Vos5, Vincenzo Di Marzo4,6,7, Serge Luquet8 & Patrice D. Cani1,*

To study the roles of intestinal N-acylethanolamines (NAEs) on metabolism in the context of obesity-associated metabolic disorders, we developed a mouse model of inducible deletion of NAPE-PLD –NAEs synthetizing enzyme– specifically in the intestinal epithelial cells (NapepldΔIEC). The deletion was induced in adult mice, that were exposed to control or high fat diet for several weeks. Body weight, fat mass and energy intake were weekly monitored; we investigated glucose metabolism and the composition of the gut microbiota.

During the first week of HFD, NapepldΔIEC mice eat significantly more than WT; the hyperphagia of KO mice is associated with reduced expression of hypothalamic anorexigenic peptides in comparison to WT mice. Upon long-term HFD, intestinal NAPE-PLD KO mice develop a worsened obesity and hepatic steatosis than WT mice; this is not associated with a modification of oral glucose tolerance and insulin sensitivity. Metagenomic analyses revealed that HFD feeding strongly changes gut microbiota composition; on the other hand, NapepldΔIEC slightly modify the gut microbiota under CT diet and HFD. Treatment with Akkermansia muciniphila, a bacterium influencing endocannabinoid system’s tone, partly counteracted the effects of the deletion.

In conclusion, our results support the hypothesis that intestinal NAPE–PLD plays a role in the regulation of whole-body energy homeostasis in response to fat intake and that additional target are involved in A. muciniphila-to-host crosstalk. Finally, considering that the intestinal NAPE-PLD deletion influences food intake after the first exposure to fat, we hypothesize that this enzyme is a key sensor involved in a gut-to-brain axis.

Keywords: Metabolism; obesity; endocannabinoid system, intestinal N-acylethanolamines

Acknowledgement: We thank A. Barrois, A. Bever, H. Danthinne, T. Duparc, B. Es Saadi, R. M. Goebbels, S. Matamoros, T. Pringels (at Université Catholique de Louvain) and C. Martin (at Université Laval), for their excellent assistance. A.E. is research associate at FRS-FNRS, Belgium. P.D.C. is a senior research associate at FRS-FNRS (Fonds de la Recherche Scientifique), Belgium. P.D.C. is the recipient of grants from the FNRS and FRSM (Fonds de la recherche scientifique médicale, Belgium). V.D. is the holder of the Canada Excellence Research Chair on the Microbiome-Endocannabinoidome Axis in Metabolic Health at Université Laval. This work was supported by the FRFS-WELBIO under grant WELBIO-CGR-2017-C2, and the Funds Baillet Latour (Grant for Medical Research 2015), ERC Starting Grant 2013 (336452-ENIGMO).

1Metabolism and Nutrition Research Group, Louvain Drug Research Institute, WELBIO, Université catholique de Louvain, 1200 Bruxelles, Belgium; 2Institut Curie, CNRS, Unité 144, 75248 Paris, France; 3Bioanalysis and Pharmacology of Bioactive Lipids Research Group, Louvain Drug Research Institute, Université catholique de Louvain, 1200 Bruxelles, Belgium; 4Quebec Heart and Lung Institute Research Centre, Université Laval, Quebec City, Canada; 5Laboratory of Microbiology, Wageningen University, 6708WE Wageningen, the Netherlands; 6Institute of Nutrition and Functional Foods, Université Laval, Quebec City, Canada; 7Endocannabinoid Research Group, Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Italy; 8Université Paris Diderot, Sorbonne Paris Cité, BFA, UMR8251, CNRS, 75013 Paris, France; 9These authors contributed equally. *Correspondence: patrice.cani@uclouvain.be

27

Implication of the gut microbiota in personalized metabolic response to dietary inulin in obese patients

by

Julie Rodriguez1, Audrey M. Neyrinck1, Tiphaine Le Roy1, Sophie Leclercq1, Sophie Hiel1, Patrice D. Cani1, Nicolas Lanthier2, Jean Paul Thissen3, Laure B. Bindels1, and Nathalie M.

Delzenne1

Aim Dietary supplementation with inulin as a prebiotic has been shown to lessen obesity and related metabolic disorders in some individuals. We have adressed the following question, by using mice transferred with the fecal microbiota from obese patients: do the characteristics of the gut microbiota in obese individuals explain the different physiological response to inulin?

Methods The gut microbiota of four-weeks old mice was depleted using antibiotics. Mice were then colonized with stools from four human obese patients (hum-ob) selected for different gut microbiota composition (alpha-diversity indexes, Bifidobacterium levels…). Conventional and hum-ob mice were then fed with a high-fat diet (HFD) during four additional weeks, supplemented or not with native inulin (Cosucra) (6-9 mice per group).

Main findings We demonstrated that antibiotics effectively depleted more than 99.8% of fecal total bacteria in mice. Hum-ob mice tended to gain a higher body weight during a HFD, compared to the conventional ones. Inulin supplementation largely reduced the body weight of hum-ob mice for only one donor (p<0.05). Inulin significantly reduced the hepatic steatosis (lipids and triglycerides content), as well as the increased adiposity induced by a HFD and/or gut microbiota transfer for two donors. Despite an increased caecal content – signing gut fermentation- observed in all humanized mice, the increase in Bifidobacterium spp. was not found for each donor suggesting that the inulin response could differ depending on the gut microbiota.

Conclusions Our work using a model of gut microbiota transfer from patients into mice highlighted the different response of humanized mice to dietary supplementation with inulin. We propose that the gut microbiota is an important component to be taken into account for personalized nutrition related to prebiotic dietary fibers in the future.

Keywords: Obesity, mice, gut microbiota, prebiotics, inulin, hepatic steatosis, Bifidobacterium spp.

Grant Support: This research is supported by the competitive cluster Wagralim from Wallonia (FOOD4GUT project, convention 1318148). Affiliation 1Metabolism and Nutrition Research Group, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium. 2 Laboratory of Hepatogastroenterology, Institut de recherche expérimentale et clinique, Université catholique de Louvain, Brussels, Belgium. Electronic 3 Pole of Endocrinology, Diabetes and Nutrition; Institut de Recherche Expérimentale et Clinique IREC, Université catholique de Louvain, Brussels, Belgium. e-mail: j.rodriguez@uclouvain.be

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Vitamin D as predictor of telomere length in children by

Carola JC Van Aart1, Nathalie Michels1, Isabelle Sioen1,2, Dries Martens3, Tim S Nawrot3,4 and Stefaan De Henauw1,5

Introduction: Telomere length is seen as marker for healthy ageing, as accelerated shortening is involved in the development of age-related diseases. Anti-inflammatory activities of vitamin D might positively influence telomere length, because inflammation could facilitate shortening. Evidence supporting this theory is mainly from studies in adults, whether vitamin D in children is a predictor of telomere length is unknown. Therefore, this study investigated the role of vitamin D on telomere length in children.

Methods: Healthy Belgian children (N=142) were followed-up between 2010-2015 (aged 4.7-10.7 years old at baseline, 50.4% boys). Serum vitamin D (25-hydroxyvitaminD) concentrations were measured at baseline and in 2015 using standardized protocols. Average relative telomere lengths were measured in blood samples using a quantitative real-time PCR method. Cross-sectional and longitudinal age and sex adjusted mixed linear regression models addressed the association between vitamin D and telomere length.

Results: Telomere length was shorter with increasing age (p=0.01). Higher vitamin D was associated with longer telomere length in boys, but not in girls. An interquartile increase in vitamin D (9.4 ng/mL) in boys was associated with a 2.5% (95% confidence interval: 0.1 to 4.9%; p=0.04) increase in telomere length. No longitudinal associations were seen between vitamin D and telomere length.

Conclusion: This study suggests that higher vitamin D concentrations might be associated

with longer telomere length in boys. These findings highlight the potential beneficial effects of vitamin D on ageing, especially since vitamin D concentrations might be easily adjustable through nutritional supplements.

Keywords: Vitamin D, Telomeres, children

1 Department of Public Health, Ghent University 2 Department of Food Safety and Food Quality, Ghent University 3 Centre for Environmental Sciences, Hasselt University 4 Department of Public Health and Primary Care, Leuven University 5 Department of Health Sciences, University College Ghent e-mail: carola.vanaart@ugent.be

29

Low 25-hydroxyvitamin D plasma levels are associated with an increased risk for acute myocardial infarction in

patients with pre-existing coronary heart disease by

Van Asbroeck S1,2*; Lysne V3; Steenackers N1; Seifert R4; Nygard O3,4; Dierkes J2,5

Background: Low 25-hydroxyvitamin D (25(OH)D) plasma levels are associated with a higher risk for cardiovascular events. However, the underlying mechanism is unidentified. To elucidate the mechanism, it is important to know the determinants of 25(OH)D levels.

Objective: To generate a multiple linear regression model of 25(OH)D determinants and explore the association between 25(OH)D levels and acute myocardial infarction (AMI) risk in patients with stable angina pectoris (SAP).

Methods: 4,116 patients with SAP were included from April 14, 1999 to April 26, 2004 and followed-up until December 31, 2006. At baseline, body mass index (BMI) and subject characteristics were assessed and an extensive blood sample analysis was performed. Dietary intake was determined in 1,981 patients. Baseline measurements were used to generate a multiple linear regression model predicting 25(OH)D levels. The effect of 25(OH)D levels on AMI risk was studied with Cox regression analysis.

Results: 25(OH)D levels were positively associated with dietary vitamin D intake, serum apolipoprotein A-1 and blood-draw during sunny months. Furthermore, 25(OH)D levels were negatively associated with smoking, BMI and estimated glomerular filtration rate. Regarding AMI risk, one standard deviation decrease in Ln-transformed 25(OH)D was associated with a 24% higher risk for AMI (p<0.001).

Conclusion: The multiple linear regression model confirmed a number of 25(OH)D determinants. One standard deviation decrease in Ln-transformed 25(OH)D was associated with a 24% higher risk for AMI in patients with pre-existing SAP.

Keywords: Vitamin D, coronary heart disease, apolipoprotein A-1

1 Clinical and Experimental Endocrinology, KU Leuven and Department of Endocrinology, University Hospitals Leuven, Belgium 2 Department of Clinical Medicine, University of Bergen, Norway

3 Department of Clinical Science, University of Bergen, Norway 4 Department of Heart Disease, Haukeland University Hospital Bergen, Norway 5 Laboratory Medicine and Pathology, Haukeland University Hospital Bergen, Norway *E-Mail: stephanie.vanasbroeck@student.kuleuven.be

30

The effect of diet in fertility patients undergoing assisted reproductive technology

by

Vermeulen A1, Boedt T, Spiessens C, Matthys C

Background: Worldwide, 8 up to 12% of the couples are facing infertility. Whereas lifestyle factors, such as poor diet, are playing an important role in the development of infertility, it is suggested that promoting healthy dietary patterns may improve fertility and possibly the outcome of fertility treatments, e.g. assisted reproductive technology (ART). Therefore, it is preliminary to investigate whether diet is associated with ongoing pregnancy rate (OPR).

Method: Between September 2016 and February 2018, a retrospective dataset of 26 female

fertility patients visiting the University Fertility Centre of Leuven was collected. These women completed a FFQ at two timepoints during their treatment. Based on the FFQ, the consumption of the main food groups was calculated and compared with the Flemish Food based Dietary Guidelines (FBDG). Paired T-tests were performed to detect changes in diet over time and logistic regression was performed to test whether diet influences the outcome of ART, defined as OPR.

Results: The overall female fertility patient does not meet the Flemish FBDGs. With an average daily consumption of 147 g, only 12% meets the recommended vegetable intake. Paired T-tests do not report dietary changes during treatment (all food groups show p>0.05). The logistic regression model, which includes solely 26 patients, does not show an association between diet and OPR.

Conclusion: Female fertility patients have poor diet quality and do not change their

consumption pattern during treatment. Whether diet affects the outcome of ART, research including larger study populations is required.

Keywords: Infertility, assisted reproductive technology (ART), ongoing pregnancy rate (OPR), food frequency questionnaire (FFQ)

1 Clinical and Experimental Endocrinology, KU Leuven E-mail: astrid.vermeulen@student.kuleuven.be

31

Stability of Potential Renal Acid Load

by

Vickà Versele1, Peter Clarys, Benedicte Deforche, Johan Lefevre, Evelien Mertens, Ruben Charlier, Sara Knaeps, Dirk Aerenhouts

The potential renal acid load (PRAL) has been described in relation to different health outcomes. (1-3) Outcomes over time and conclusions made are often based on baseline dietary intake values. (4,5) However, to study reliable long-term associations, parameters calculated based on dietary intake data such as PRAL, must be stable over time. Therefore, the aim of this study was to assess the stability of PRAL and its components over a 10 year time period.

Methods: PRAL values of 3-day dietary intake data from 197 women and 373 men on two assessment moments (2002 – 2004 and 2012 – 2014) were calculated. Pearson correlation and intra-class correlation coefficients were used for assessing the stability of PRAL and its components. Level of agreement between the two assessment moments was estimated after splitting up subjects in quintiles of PRAL, calculating kappa values and changes of quintiles over time.

Results: No significant differences in mean PRAL over time were found. Stability of PRAL and its components was low. Poor agreement between first and second assessment was shown by low kappa values and change of most of the subjects to an adjacent and non-adjacent quintile after 10 years.

Conclusion: Stability of PRAL over a 10 year time period could not be confirmed based on nutrition assessment carried out by 3-day dietary records, even though no significant difference between mean PRAL and its components over time was found. Therefore, interpretation of longitudinal outcomes based on PRAL and its component calculated at baseline should be interpreted with caution.

32

TABLE 1. Mean (SD) Potential Renal Acid Load (PRAL), correlation coefficients of PRAL, and percentage of

subjects in the same quintile, adjacent or non-adjacent quintile after 10 year

Men (n=373) Women (n=197)

Mean (SD) PRAL 2002 – 2004 2012 –

2014 p 2002 – 2004 2012 – 2014 p

7.3 (19.5) 6.2 (17.7) 0.327 0.8 (14.8) -0.3 (17.3) 0.467

Correlation coefficients of PRAL between 2002 – 2004 and 2012 – 2014

Pearson r p ICC p Pearson

r p ICC p

0.358 <0.001 0.356 <0.001 0.168 0.019 0.165 0.01

Percentage of subjects in the different quintiles after 10 year

Same category 25.5% 28.1%

Adjacent category difference 35.8% 30.6%

Non-adjacent category difference

38.7% 41.3%

Keywords: nutrition assessment, potential renal acid load, stability

1 Faculty of Physical Education and Physiotherapy, Department of Movement and Sport Sciences, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium E-mail: Vicka.versele@vub.be

(1) Xu H, Akesson A, Orsini N, et al. Modest U-Shaped Association between Dietary Acid Load and Risk of All-Cause and Cardiovascular Mortality in Adults. The Journal of nutrition. 2016;146(8):1580-5.

(2) Han E, Kim G, Hong N, et al. Association between dietary acid load and the risk of cardiovascular disease: nationwide surveys (KNHANES 2008-2011). Cardiovascular diabetology. 2016;15(1):122.

(3) Luis D, Huang X, Riserus U, et al. Estimated dietary acid load is not associated with blood pressure or hypertension incidence in men who are approximately 70 years old. The Journal of nutrition. 2015;145(2):315-21.

(4) Alexy U, Remer T, Manz F, Neu CM, Schoenau E. Long-term protein intake and dietary potential renal acid load are associated with bone modeling and remodeling at the proximal radius in healthy children. The American journal of clinical nutrition. 2005;82(5):1107-14.

(5) Pedone C, Napoli N, Pozzilli P, et al. Quality of diet and potential renal acid load as risk factors for reduced bone density in elderly women. Bone. 2010;46(4):1063-7.

33

Cellular quercetin accumulation and localization under stressed conditions

by

H. Vissenaekens1,a,b, C. Grootaert 2,a, G.B. Gonzales3,a,b, K. Raes4,a, T. Van de Wiele5,c, G. Smagghe6,b, A. Rajkovic7,a, J. Van Camp8,a

Aim: The aim of this study is to investigate flavonoid uptake in several cell lines using innovative detection techniques. Moreover, changes of cellular uptake, accumulation and localization of quercetin, the most abundant flavonol in the daily diet, as affected by treatment with valinomycin (model for the emetic toxin cereulide) as a stressor are studied.

Approach: After cytotoxicity studies, the intestinal HCT116 and Caco-2 cells, as well as the endothelial EA.hy926 cells were exposed to quercetin and/or valinomycin. Subsequently, the changes of cellular quercetin uptake were investigated using (i) an innovative in-house developed spectrofluorometric method, (ii) an in-house developed high throughput flow cytometric method and (iii) confocal microscopy.

Results: Intracellular quercetin accumulation was detected by all three, of the previous described, techniques in all cell lines. Significant higher quercetin accumulation was observed when the cell lines were stressed by valinomycin treatment. Confocal microscopic results showed that quercetin (metabolites) were localized in the cell membrane of the Caco-2 cells under non-stressed conditions, whereas the valinomycin stress induced the migration of the compounds into the cytoplasm of the Caco-2 cells.

Conclusion: This study shows that the in-house developed spectrofluorometric and flow cytometric method can be used to detect quercetin uptake at the cellular level and that there is a tendency towards an altered localization and accumulation of quercetin (metabolites) in intestinal cells when stressed by valinomycin.

keywords: in vitro cell cultures, flavonoids, cellular stress, spectrofluorometry

1 Hanne.Vissenaekens@UGent.be; 2 Charlotte.Grootaert@UGent.be; 3 Gerard.Gonzales@UGent.be; 4

Katleen.Raes@UGent.be; 5 Tom.VandeWiele@UGent.be; 6 Guy.Smagghe@UGent.be; 7

Andreja.Rajkovic@UGent.be; 8 John.VanCamp@UGent.be

a Department of Food technology, Safety and Health, Faculty of Bioscience Engineering, Ghent University; b

Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University; c Department of Biotechnology, Faculty of Bioscience Engineering, Ghent University

H. Vissenaekens is an SB PhD fellow at FWO (1S18417N). Special thanks to the Special Research Fund (BOF) of Ghent University for financing the Ph.D. of GB Gonzales. All authors would like to than the Special Research Fund of Ghent University (BOF 01B04212) for the financial support.

34

A 10-year longitudinal study on the associations between changes in plant-based diet indices,

anthropometric parameters and blood lipids in a Flemish adult population

by

Jana Waterplas, Vickà Versele, Eva D’Hondt, Benedicte Deforche, Johan Lefevre, Evelien Mertens, Ruben Charlier, Sara Knaeps, Peter Clarys

Background: In the context of sustainability and health, plant-based diets are recommended.(1,2) Recently, different plant-based indices have been developed. (3,4) However, not all plant foods can be considered as beneficial, therefore a distinction needs to be made between healthful and unhealthful plant-based foods.

Objective: The aim of this study was to apply a plant-based diet index (PDI), a healthful plant-based diet index (hPDI) and an unhealthful plant-based diet index (uPDI) on a representative population and investigate the longitudinal association between changes in these indices with changes in anthropometric parameters and blood lipids.

Methods: Three day diet records completed in 2002-2004 and 2012-2014 of 352 men and 190

women were available for analysis. Waist circumference (WC) and Body Mass Index (BMI) were used as anthropometric parameters. Three indices (PDI, hPDI and uPDI) were calculated. Associations between changes in dietary indices and changes in anthropometric parameters and blood lipids were tested using multivariate linear regression. The analyse was adjusted for potential confounders.

Results: In women an increase PDI was associated with an increase in BMI. In men an

increase in uPDI was associated with an increase in the anthropometric parameters. No longitudinal associations were found between the dietary indices and blood lipids, except for the positive association between uPDI and total cholesterol in women.

Conclusion: Mean index values did not differ over time. Only few associations were found between dietary indices and anthropometric parameters and blood lipids.

(1) Van Loo et al. , sustainable and plant-based eating: Perceived (mis)match and involvement-based consumer segments as targets for future policy. Food Policy [Internet]. 2017;69:46–57. (2) Lacour C et al. Environmental Impacts of Plant-Based Diets: How Does Organic Food Consumption Contribute to Environmental Sustainability? Front Nutr [Internet]. 2018;5(February):1–13. (3) Martinez-Gonzalez MA, Sanchez-Tainta A, Corella D, Salas-Salvado J, Ros E, Aros F, et al. A provegetarian food pattern and reduction in total mortality in the Prevencion con Dieta Mediterranea (PREDIMED) study. Am J Clin Nutr. 2014;100(Supplement_1):320S–328S. (4) Satija et al. Healthful and Unhealthful Plant-Based Diets and the Risk of Coronary Heart Disease in U.S. Adults. J Am Coll Cardiol. 2017;70(4):411–22.

Keywords: Plant-based diet index, Unhealthful plant-based diet index, Healthful plant-based diet index, Waist circumference, Body mass index, Blood cholesterol

1 Faculty of Physical Education and Physiotherapy, Department of Movement and Sport Sciences, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium Jana.Waterplas@vub.be

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TABLE 1. Associations between changes in dietary indices and changes in anthropometric parameters (waist circumference, Body Mass Index) and total cholesterol (TC)

Men (N = 352)

Women (N = 190)

Men (N = 352)

Women (N = 190)

2002-2004 2012-2014 2002-2004 2012-2014 Model 1 Model 2 Model 1 Model 2

β R² β R² β R² β R²

PDI (mean ± SD) 53.66 ± 5.82 53.89 ± 5.89 53.73 ± 5.62 53.84 ± 5.56

uPDI (mean ± SD) 53.89 ± 6.46 53.86 ± 6.68 55.43 ± 5.54 54.81 ± 5.82

PDIa – BMI -0.028 -0.002 0.034 0.017 0.142 0.015 0.135* 0.219

uPDIb – WC 0.204*** 0.039 0.153 0.080 0.011 -0.005 0.000 0.217

uPDIb – BMI 0.144** 0.018 -0.049 0.018 0.015 -0.005 -0.001 0.200

uPDIb – TC 0.036 -0.002 -0.185 -0.023 0.148* 0.016 0.042 0.166

*p < 0.05, **p < 0.01, ***p < 0.001 Model 1: unadjusted Model 2: adjusted for age, residual change score smoking, residual change score VO2peakrelative and residual change score waist circumference for the association with blood lipids.

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Potential functionality of protein hydrolysates for glycaemia control

by

Virgilio N.1, Vitaglione P.1

Dietary proteins may contain some bioactive peptides (BAPs) encrypted within their primary structures. BAPs may be delivered in the food during processing and/or in the gastro-intestinal tract during food digestion and can exert some biological effects beyond nutrition such as antimicrobial, anti-thrombotic, antihypertensive, opioid and immunomodulatory effects. BAPs modulating blood glucose response are promising ingredients for functional foods development. They work through inhibition of the enzyme dipeptidyl peptidase IV (DPP-IV) that modulate glucose homeostasis by cleaving glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide. The aim of this study was to evaluate the potential activity of casein (CH) and soy (SH) protein hydrolysates as well as of CH and SH enriched biscuits (CHB and SHB) on post-prandial glucose response in vitro. Control biscuits (ConB) without protein hydrolysates and two types of CH and SH-enriched biscuits providing 4.5% (CHB1 and SHB1) and 13% (CHB2 and SHB2) of each hydrolysate were developed. CH, SH, CHB1, CHB2, SHB1 and SHB2 were subjected to in vitro simulated gastrointestinal digestion and the ability of the digested samples to inhibit DPP-IV activity was assessed. In vitro glycaemic index (GI) of the biscuits was also measured. Data showed that CH and SH behaved as mixed and competitive inhibitor of DPP-IV with an IC50 of 2.59 mg/ml and 3.56 mg/ml (p<0.05), respectively, when tested alone. No significant difference between digested biscuits for the inhibition of DPP-IV activity was observed. The GI of the biscuits was in the order ConB> CHB1>SHB1>CHB2>SHB2. This study suggested that CH and SH maybe functional ingredients for glycaemia control through inhibition of DPP-IV activity. A food matrix effect could hide the bioactivity of CH and SH at the doses used in the biscuits during in vitro enzymatic digestion.

Bioactive peptides; dietary protein; dipeptidyl peptidase IV; functional food; glycaemia control; simulated gastrointestinal digestion.

1 Department of Agricultural Sciences, University of Naples “Federico II”, Portici, Italy nicolina.virgilio@unina.it

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Dietary intake of polyphenols in European adolescents: the HELENA study

by

Ratih Wirapuspita Wisnuwardani1,2, Nathalie Michels1, Stefaan De Henauw1

Purpose Knowledge about polyphenols intakes and their determinants among adolescents might be helpful for planning targeted prevention strategies at an early age.

Methods In the European multicenter cross-sectional HELENA study of 2006-2007, 2428 subjects (47 % boys) had data on dietary intake of polyphenols from two non-consecutive 24 hour recalls via linking with the Phenol-Explorer database. Differences by sex, age, country, BMI, maternal education, paternal education, family affluence, smoking status, alcohol use, and physical activity were explored by linear regression.

Results Median, lower and upper quartiles of polyphenol intakes were 326, 167 and 564 mg/d, respectively. Polyphenol intake was significantly higher in the oldest (16-17.49 years), girls, non-Mediterranean countries, lowest BMI, highest maternal education, and alcohol consumers. Main food contributors were fruit (22.9 %, mainly apple and pear i.e. 16.3 %); chocolate products (19.2 %); and fruit and vegetable juices (15.6 %). Main polyphenol classes were flavonoids (74-76 % of total) and phenolic acids (18-20 % of total). The three most consumed polyphenols were hesperidin, proanthocyanidin polymers (>10 mers), and proanthocyanidin 4-6 oligomers.

Conclusion The current study provided for the first-time numbers on the total polyphenol intake and their main food sources in a heterogeneous group of European adolescents. Major differences with adult populations are the lower polyphenol consumption and some unhealthy major food group contributors (chocolate, biscuits) instead of vegetables and coffee. The discussed determinants and polyphenol types already point to some important population groups that need to be targeted in future public health initiatives.

Keywords: Polyphenol · Consumption · Food contributors · HELENA · Determinants · Mediterranean

1 Nutrition and Food Security Unit, Public Health Department, Medicine Faculty, Ghent University, Ghent, Belgium

2 Public Health Faculty, Mulawarman University, East Kalimantan, Indonesia

RatihWirapuspita.Wisnuwardani@Ugent.be; ratih@fkm.unmul.ac.id

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Fig. 1 Daily mean polyphenol intake (mg/day) and percentage contribution, stratified by sex (a boys; b girls) and center, adjusted for age and examination month.

Mean energy-adjusted polyphenol intake was 204mg/1000kcal. The highest total polyphenol intake was in Ghent (in boys 588 mg/day) and in Vienna (in girls 508 mg/day), whereas the lowest intake in both sexes was in Zaragoza (in boys 212 mg/day and in girls 203 mg/day) (Figure 1).