Metabolomics for monitoring dietary exposure

Lead Research Organisation: Aberystwyth University
Department Name: IBERS

Abstract

There is strong evidence that dietary choices modulate risk of developing a range of chronic diseases and strategies for reducing disease burden emphasize the importance of changing dietary patterns. Current public health guidelines have common themes that encourage consumption of specific food groups, for example, oily fish, wholegrain foods, fruits and vegetables whilst reducing intakes of fatty and sugary foods. To maximise the effectiveness healthy eating interventions in the UK (and elsewhere), we need validated population screening methods with which to measure food consumption before, during and after the intervention. Conventional dietary survey methods are a considerable burden for study participants and are subject to bias. In addition, the processing and analysis of dietary data is extremely labour intensive with the net result that such dietary survey information may be difficult to validate. Such difficulties limit the ability to interpret the outcomes of healthy eating interventions.

Significant advances have been made recently in developing 'biomarkers' of dietary intake based on metabolites found in urine or blood but, currently, chemical biomarkers are available for only a relatively small number of specific foods and food components and most are of uncertain validity.

Funded largely by the UK Food Standards Agency the research teams in Aberystwyth, Newcastle and Imperial College have used metabolomics approaches to develop a novel strategy for discovery of new biomarkers of food intake. "Metabolomics" describes the measure of all (or many) of the metabolites (small molecules) in biological fluids such as blood or urine. Following digestion, absorption and metabolism, foods give rise to thousands of different metabolites in the human body and the appearance of certain metabolites is characteristic of particular foods. In our earlier work, we have developed urine sampling methods potentially suitable for use in population studies and have explored the use of comprehensive, non-targeted chemical analysis of urine (metabolomics technology) to identify dietary biomarker leads. Using this approach, we have identified potential biomarkers for several foods of high public health significance including oily fish such as salmon, citrus fruit, other fruit such as raspberries and the green vegetable broccoli. We thus now intend to test the hypothesis that the chemical composition of urine samples from free-living individuals will accurately reflect dietary intake.

The research programme will have the following major aims:

1. To identify chemicals in urine associated with recent consumption of specific foods deemed to have high public health importance.

2. To demonstrate a quantitative relationship between the amount of each specific food eaten and each potential chemical 'biomarker' in urine samples.

3. To validate a simple urine sampling procedure that is minimally intrusive to the every day activities of individuals participating in dietary surveys.

4. To assess the performance of dietary exposure biomarkers in the prediction of habitual diet in the UK.

The research strategy will be designed to integrate where possible with major surveys of UK dietary habits which will offer future opportunity to determine whether the adoption of biomarker technology improves the cost effectiveness and accuracy of population-level dietary surveys. In addition there will be distinctive advantages in future longitudinal studies aiming to generate definite links between disease risk and individual habitual exposure to specific foods. The results of this research will provide data to allow statuary bodies to refine dietary advice provided to consumers as well as offering further opportunity to interact with food producers, manufacturers and retailers to encourage provision of foods targeted to improve the overall health of the UK population.

Technical Summary

In collaboration with colleagues at the MRC Human Nutrition Research, we have identified a panel of foods of high public health importance suitable for exposure monitoring in urine using biomarker technology. Free-living volunteers will consume these specific foods as part of everyday eating patterns over 3 days before collection of urine and blood for identification of potential biomarkers.

The timings of specific meals and of biofluid collection will be pre-determined to facilitate investigation of the duration of food-derived metabolite signals and the most informative urine collection strategies. In addition, 400 study participants consuming their usual diets will be recruited. We will estimate habitual dietary intake using the EPIC FFQ and collect "spot" urine samples. This population sample will be used to test the utility of putative metabolite biomarkers of dietary intake.

NMR and LC-MS technology will be used to generate comprehensive metabolite profiles of urine samples. Supervised data mining methods will identify signals explanatory of exposure to specific foods. Targeted NMR and accurate mass FT-ICR-MS will be used to structurally annotate metabolites with biomarker potential. Targeted analytical methods will be established to simultaneously measure several biomarker signals (e.g. using MRM approaches) in urine.

In controlled residential food intervention studies, volunteers will be asked to consume known amounts of test foods for which we have identified potential biomarkers and urine and blood will be collected at defined intervals for up to 3 days after consumption. We aim to describe the kinetics of signal decay and to determine the quantitative relationships between food intake and urinary metabolite concentrations.

Biomarkers proving informative will be tested for utility using dietary data and urine samples from both recent National Diet and Nutrition Survey studies (24hr urines) and the 400 "spot" urines from the present study.

Planned Impact

Who will benefit from this research?

This proposal aims to help future studies to investigate more efficiently the challenging question of the effect of dietary exposure on disease prevention - health - or causation. We therefore anticipate that this research will have a wide range of beneficiaries (in addition to the academic community) including:

- Regulators with responsibilities in the area of food and health e.g. the European Food Safety Authority and the UK's Advisory Committee on Novel Foods and Processes,
- The food industry including those developing novel foods or food ingredients aimed at personalised nutrition and those advising consumers,
- Manufacturers of analytical platforms and software tools for metabolomics analyses,
- The general public.


How will they benefit from this research?

Regulators with responsibilities in the area of food and health are currently exerting considerable effort to get across the message to consumers about the benefit to health and well being of including, for example, sufficient amounts of fruit and vegetables (F & V) in their diet. However, their effort is pre-empted by the conflicting conclusions resulting from current epidemiological and dietary intervention studies in humans. Within the next 4-5 years this project will provide food biomarker technology that will allow future researchers to generate objective quantitative measures of habitual dietary exposure at the population level in the UK. Quicker and cheaper high throughput methods to assess dietary exposure will support the scale up of longitudinal population surveys to allow links to be made between exposure to specific food and future health risks.

Thus any health outcomes associated with dietary trends can allow regulators to develop even stronger messages to persuade the general public to change eating habits.

It is anticipated that there would be incremental integration of biomarker technology into population studies, starting with validation of the present dietary recording tools, particularly for monitoring accurately exposure to the individual components of composite meals. As the 'coding' of meals from diet diaries is extremely time consuming (as well as a source of much experimental inaccuracy) it is expected that the availability of validated food biomarker technology would allow organisations responsible for diet habit monitoring to simplify the data collection protocols, making the surveys more cost effective.

Outcomes from this research will provide the food industry with more robust measures and tools for R&D in the area of personalised nutrition and it is anticipated that impact will be realised within the next 5 years.

A deep understanding of metabolites in urine will help manufacturers of mass spectrometers and NMR instruments to develop new data processing and data annotation software tools more adapted to human nutritional metabolomics.

The general public are expected to benefit from this project in at least 3 ways i) through provision of objective information about the complex interactions of humans with their diet; ii) by enhancing the quality and robustness of evidence which will be required to elaborate and support dietary recommendation on consumption of healthy foods, and iii) by encouraging the food industry to focus on the marketing of food products with proven health benefits.

What will be done to ensure that they have the opportunity to benefit from this research?

We will disseminate the outcomes from the research project via the conventional route of academic publications using open access journals where possible. This will ensure widespread access by all stakeholders to the project outcomes following the important quality assurance step of peer review.
In addition, we will present findings on an on-going basis via conference presentations targeting conferences and workshops attended by diverse user groups including industry and regulators.

Publications


10 25 50
Beckmann M (2013) Dietary exposure biomarker-lead discovery based on metabolomics analysis of urine samples. in The Proceedings of the Nutrition Society
 
Description EiT-Health Innovation Coordination Committee
Geographic Reach Europe 
Policy Influence Type Membership of a guideline committee
URL https://eithealth.eu/
 
Description MRC/NIHR event for OSCHR: Review of Nutrition and Human Health Research - Workshop 7th & 8th July 2016, London
Geographic Reach National 
Policy Influence Type Participation in a national consultation
 
Description Direct support for EIT Health Nutrition and Healthy Living Partnering Workshop
Amount £3,000 (GBP)
Organisation Welsh Government Food and Drink 
Sector Public
Country Unknown
Start 05/2015 
End 05/2015
 
Description EIT-Health Innovation by Design Project: Cook 2 Health
Amount € 257,116 (EUR)
Funding ID EIT-Health 2016 
Organisation European Institute of Innovation and Technology (EIT) 
Sector Public
Country Hungary, Republic of
Start 03/2016 
End 12/2017
 
Description EIT-Health Innovation by Design Project: FACET
Amount € 188,619 (EUR)
Funding ID EIT-Health 2016: 1.1.3 
Organisation European Institute of Innovation and Technology (EIT) 
Sector Public
Country Hungary, Republic of
Start 02/2016 
End 12/2017
 
Description Endeavour Grant: for EIT Health Nutrition and Healthy Living Partnering Workshop
Amount £3,000 (GBP)
Organisation Life Sciences Research Network Wales 
Sector Academic/University
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 05/2015 
End 05/2015
 
Description European Structural Funds programmes 2014-2020 West Wales and the Valleys
Amount £20,500,000 (GBP)
Organisation European Commission (EC) 
Department European Regional Development Fund (ERDF)
Sector Public
Country European Union (EU)
Start 12/2014 
End 11/2018
 
Description Knowledge Economy Skills Scholarships
Amount £68,000 (GBP)
Organisation European Commission (EC) 
Department European Social Fund
Sector Public
Country European Union (EU)
Start 07/2016 
End 06/2019
 
Description Research and Innovation Campus
Amount £12,000,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 12/2014 
End 11/2018
 
Description Fecal derivative of wholegrain diet 
Organisation University of Bologna (Università di Bologna UNIBO)
Country Italy, Italian Republic 
Sector Academic/University 
PI Contribution Trained a visiting Ph.D student (Miss Danielle Taneyo - 4 months) in metabolomics technology to discover dietary biomarkers
Collaborator Contribution Provided samples from a wholegrain dietary intervention
Impact Ph. D thesis
Start Year 2013
 
Description Fruit and Vegetable consumption level biomarker discovery 
Organisation Queen's University of Belfast
Department School of Medicine, Dentistry and Biomedical Sciences
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Used metabolomics methods to discover potential biomarkers of Total Fruit and Vegetable Exposure level in urine. My team helped to design the urine sampling method and experimental statistical design. Trained a visiting Post doc in metabolomics methods
Collaborator Contribution Undertook a Fruit and Vegetable dietary intervention as part of an MRC grant. Collected dietary exposure data and specifically timed urine samples.
Impact Potential biomarkers are currently under structural analysis. Collaboration is multidisciplinary linking Nutrition Research in Belfast with Metabolomics and Machine Learning in Aberystwyth
Start Year 2014
 
Description Standardised MRM tests for dietary exposure biomarker quantification 
Organisation University of Barcelona (UB)
Department Faculty of Pharmacy
Country Spain, Kingdom of 
Sector Academic/University 
PI Contribution Identified Meat and Fish biomarkers and quantification methods
Collaborator Contribution Identified Fruit and Veg Biomarkers and quantification methods
Impact We have joined a EIT-Health consortium (Cook 2 Health) which funds our collaboration
Start Year 2015
 
Description Tyrosine metabolism in humans 
Organisation Aberystwyth University
Department Sport and Exercise Science
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Developed methodology to measure tyrosine metabolism in human plasma and analysed samples in an intervention study
Collaborator Contribution Organised a Tyrosine supplementation cohort and measured exercise capacity in humans
Impact Two publications journal articles
Start Year 2012
 
Description UKB CMC - UCL collaboration 
Organisation University College London (UCL)
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution UK Biobank Cardiometabolic Consortium coordination and leadership
Collaborator Contribution Working with Prof Harry Hemingway and colleagues through the Consortium's CHD working group with a focus on exploring opportunities for large scale CVD research arising from linking electronic health records .
Impact collaboration in progress
Start Year 2012
 
Description BBC 1 Wales Television interveiw 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Media (as a channel to the public)
Results and Impact Describing the science behind chemical analysis of urine to monitor diet

Invitations to write articles for trade magazines in the Food and Drink sector
Year(s) Of Engagement Activity 2013
 
Description BBSRC Institutes Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact National Institutes of the Biosciences, 2013. 18-20th June 2013. Roslin Institute, Edinburgh. Title "Detecting the dietary component of the human 'exposome' " Over 150 people attended event including newpaper and other media representatives.


Several UK researcher expressed interest in future collaboration , including the BBSRC Chief Executive.
Year(s) Of Engagement Activity 2013
 
Description Bioinformatics Awareness 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact Made researchers aware of the impact of using Machine Learning and HPC technology to process and interrogate complex MS data

Several Ph.D students and postdocs keen to learn how to use software. This increased the visibility of the Diet and Health group within the Institute and possibly enabling future discussions
Year(s) Of Engagement Activity 2014
URL https://docs.google.com/document/d/1MBBY28ICdAKZh3YdxhWS1fJ0XCI0hBn8poK7pfmB_-s/edit?pli=1
 
Description British Geriactric Society meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact Presentation to outline urine test to screen for protein intake in pre-frail individuals
Year(s) Of Engagement Activity 2016
URL https://online.bgs.org.uk/events/regions/walesspring16
 
Description EIT Health "Dietary Impact on Healthy Aging Phenotype" Partnering Workshop, Cardiff 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Organised a workshop with support of Welsh Government Food and Drink to raise awareness of EIT-Health and get companies involved in translational projects. From this we attracted 2 EIT-Health consortia and a KESS studentship as well as stimulating further interest from Welsh Government in supporting our Institute research strategy.
Year(s) Of Engagement Activity 2015
 
Description EIT-Health Partnership meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Activity centred on developing research consortia for translation of research relevant to health sector. My input stimulated ideas relating to human phenotyping.
Year(s) Of Engagement Activity 2016
URL http://eithealth.eu/innovation-by-design-8th-march-in-leuven/
 
Description EIT-Health 2016 Summit in Barcolona 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Provided slides for two talks. Had the opportunity to present two posters and discuss translational projects with a large number of meeting participants.
Year(s) Of Engagement Activity 2016
URL https://eit.europa.eu/interact/events/eit-health-summit
 
Description EIT-Health matchmaking meeting in Warsaw 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact The meeting aimed to provide structured discussion opportunities between Draper and European companies interested in urine biomarker technologies
Year(s) Of Engagement Activity 2016
URL http://eithealth.eu/innovation-by-design-matchmaking-event-in-warsaw/
 
Description Food Bioactives and Health Conference, Norwich, UK 13-15th September 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Oral Presentation: Dietary exposure biomarkers and metabolic profiling technology to support a standardised validation pipeline for functional foods
Poster Presentation: Developing community-based urine sampling methods to facilitate dietary exposure biomarker technology for population assessment.
Poster Presentation: Quantification of dietary biomarkers in spot urine samples reflects the intake of foods of UK high public health importance
Year(s) Of Engagement Activity 2016
URL http://www.ifr.ac.uk/news/latest-news/2016/06/food-bioactives-and-health-conference-september-2016/
 
Description Food Standards Agency - Horizon Scanning 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/parliamentarians
Results and Impact Food Standards Agency very interested in food analysis methodology

Second Higher level meeting now requested for later in year
Year(s) Of Engagement Activity 2014
 
Description Frailty workshop in Madrid 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Outreach meeting to inform stakeholders supporting frail individuals about use of urine biomarkers to monitor nutritional status
Year(s) Of Engagement Activity 2016
 
Description Human Nutrition Research visit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Presentation to HNR staff and Epidemiology Groups in Cambridge
Year(s) Of Engagement Activity 2016
 
Description InnoLife Summit - Erlangen 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact John Draper was asked to act as one of the Innovation Co-coordinators for the InnoLIFE consortium, representing 6 EU countries. The summit aimed to develop thematic areas for future collaborative work in EU between Industry, Academics, Health Care Organisations and Government Departments
Year(s) Of Engagement Activity 2015
 
Description LabTube Video mage at 2015 Metabomeeting in Cambridge 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Invited to make a video describing my research presented as a Keynote speaker at the 2015 Metabomeeting in Cambridge. This is web accessible on LabTube
Year(s) Of Engagement Activity 2015
URL http://www.labtube.tv/video/labtube-meets-john-draper-aberystwyth-university
 
Description NugoWeek 2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact AJ Lloyd made an oral presentation in the Applied Omics in the field of Nutrigenomics session at NuGOweek 2014. Talk was well received and stimulated discussion in relation to sampling humans in epidemiological context

JD invited to collaborate with Portuguese National Dietary study in
Year(s) Of Engagement Activity 2014
URL http://www.nugo.org/everyone/42673/5/0/30
 
Description Nutrition Society Meeting in Dublin 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact My team provided slides derived from the MRC Programme Grant to 2 plenary speakers (Prof. John Mathers and Prof. Jayne Woodside) and also presented 2 posters on our work
Year(s) Of Engagement Activity 2016
 
Description Nutritional Metabolomics Workshop, Glasgow 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Workshop Title: "The food metabolome and biomarkers
for dietary exposure" 4-5th July 2012, Glasgow. Draper made a short presentation to set the scene and acted as acted and Facilitator and Rapporter for the session "Tools and resources for metabolite identification". Around 65 experts in epidemiology and nutritional metabolomics attended the workshop with sparked lengthy discussion on the use of bio markers to monitor dietary exposure.

The workshop Facilitators were invited to write up the proceedings and conclusions of the meeting in the form of a substantial review/position paper for the top ranked journal in the area of nutrition (American Journal of Clinical Nutrition)
Year(s) Of Engagement Activity 2013
URL http://www.nugo.org/everyone/42461/7/0/30
 
Description OSCHR reveiw of Nutrition Research (London) 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Policymakers/politicians
Results and Impact Invited participant in a meeting organised by NHOR and MRC to review the status of nutrition research in the UK. I provided slides for one of the panel members (John Mathers) and was able to ask a range of questions after different presentations and sparked discussion of the use of urine biomarkers to assess diet.
Year(s) Of Engagement Activity 2016
 
Description Presentation to BRC Nutrition in Southampton 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Presentation on biomarker technology in nutrition research
Year(s) Of Engagement Activity 2016
 
Description Presentations on Diet and Health research at national meetings of EIT-Health (Spain, UK, France, Belgium, Germany) in 2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact John Draper presented on a theme of Diet and Health research translation opportunities to national meetings of organisation that eventually came together to form EIT-Health in 2016
Year(s) Of Engagement Activity 2015
 
Description Radio Wales Interview BBC in August 2012 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Media (as a channel to the public)
Results and Impact Radio Interview for BBC Radio Wales describing the concept of urine testing to monitor recent diet

BBC Television asked for an interveiw
Year(s) Of Engagement Activity 2012
 
Description Royal Society of Chemistry Meeting: Advances in the Chemical Analysis of Food, Burlington House, London 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Oral Presentation: De novo compositional analysis of meal components: using LC-MS/MS to link food components with urinary metabolites and human health in nutritional epidemiology studies.
Year(s) Of Engagement Activity 2016
 
Description Thermo Scientific LCMS Annual User Meeting, Welcome Centre, London UK, June 2016 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Invited Oral Presentation: High Resolution LC-MS in Nutritional Metabolomics,
Year(s) Of Engagement Activity 2016
 
Description Urine Biomarker technology presented at the Medi-Wales annual meeting in Cardiff 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Presented talk on likely outputs of MRC project that could be commercialised in the future to develop point of care diagnostics. Lot of industy interest and several companies followed up afterwards.
Year(s) Of Engagement Activity 2015
 
Description Urinomics 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Conference Title: 1st International Conference on Urine Omics. 9-11 September, 2013. Talk title: " Urine sampling in human populations and discovery of dietary exposure biomarkers". Presentation sparked more then 15 mins of discussion on urine biomarker technology.

Draper asked to co-organise a session in Urinomics 2014
Year(s) Of Engagement Activity 2013
URL http://www.urinomics2013.com/
 
Description Urinomics 2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Made 2 presentations on dietary exposure biomarkers and method for urine sampling in community settings. There was considerable interest in the latter from teams involved in kidney disease research.
Year(s) Of Engagement Activity 2015
 
Description Wales Kidney Research Unit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Meeting in Bangor to discuss urine biomarker technology and application sin kidney research
Year(s) Of Engagement Activity 2016