SUPPLEMENTARY MATERIALS

ContentsAppendix I: Methods section.................................................................................................................2

Table S1. Biomarkers analysed from Dry Blood Spots in the Food4Me POP study........................2

Table S2. Genetic loci used for deriving genotypic-based personalised nutrition advice in the Food4Me POP study......................................................................................................................3

Appendix II: Example of a participant’s report......................................................................................4

Description of the personalised report provided to participants randomised to level 1, 2 and 3 of personalised nutrition.......................................................................................................................4

Table S3. Section include in the participants feedback report by intervention arm......................4

Figure 1. Example of ‘A message from the nutritionist’ (Section 1)...............................................5

Figure 2. Example of ‘How your diet compares to recommendations’ feedback (Section 2).........5

Figure 3. Example of ‘Your Physical Characteristics’ feedback (Section 3)....................................6

Figure 4. Example of ‘Your Physical Activity level’ feedback (Section 4).......................................7

Figure 5. Example of ‘Your Nutrient Profile’ feedback (Section 5).................................................8

Figure 6. Example of ‘Your Blood Profile’ feedback (Section 6).....................................................9

Figure 7. Example of ‘Your Genetic Profile’ feedback (Section 7)................................................10

Figure 7. Example of ‘Your Personalised Nutrition Advice’ feedback (Section 9).........................11

Example of advice provided to participants randomised to L0 “Control group”.............................12

Table S4. No-personalised dietary eating guidelines...................................................................12

Appendix III: Nutrient Gradations and cut-off point for deriving personalised advice........................15

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Appendix I: Methods section

Table S1. Biomarkers analysed from Dry Blood Spots in the Food4Me POP study

Fatty acid biomarkers Carotenoids biomarkers Lauric acid Fb-Fatty acids Lutein Myristic acid Zeaxanthin Pentadecyclic acid beta-kryptoxanthin Palmitic acid alpha-Caroten Palmitoleic acid beta-Caroten Margaric acid Lycopene Linoleic acid Fb-Carotenoids Diabetes biomarker Oleic acid n9 Glucose cis-Vaccenic acid Arachidic acid Lipids biomarker Gamma-linolenic acid (GLA) Cholesterol (total) Alpha-linolenic acid (ALA) Eicosenoic acid Vitamin biomarker * Eicosadienoic acid vitamin 25-OH D2 Behenic acid vitamin 25-OH D3 Arachidonic acid (ARA) Tricosylic acid Eicosapentaenoic acid (EPA) Lignoceric acid Nervonic acid Docosapentaenoic acid (DPA) Docosahexaenoic acid (DHA) Dihomo-gamma-linolenic acid (DGLA) Stearic acid Trans Fatty Acids

All biomarkers were analysed by Vitas (Vitas Ltd, Norway) except from those denoted with an *, which were analysed by DSM (DSM Nutritional Products Ltd. Switzerland). The selection of biomarkers analysed in the Food4Me study was determined by the availability of assays which could be undertaken using dry blood spot cards.

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Table S2. Genetic loci used for deriving genotypic-based personalised nutrition advice in the Food4Me POP studyGene SNP Risk Variant Dietary or Phenotypic outcomeFTO rs9939609 AA & AT BMI, Weight, Waist circumferenceFADS1 rs174546 CC Omega-3TCF7L2 rs7903146 TT & CT Fat IntakeApoE(e4) rs429358/rs7412 Haplogroups

CC/CCCT/CTCT/CCCC/CT

Saturated Fat

MTHFR rs1801133 TT & CT FolateThese 5 genetic variants were used as a basis for personalised nutritional advice for participants randomised to Level 3 of the Food4Me intervention.

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Appendix II: Example of a participant’s report

Description of the personalised report provided to participants randomised to level 1, 2 and 3 of personalised nutrition. Table S3 provide a summary of the information provided to each intervention arm. These reports were sent to participants at baseline (month 0), and at months 1, 2 and 3. Examples of each of the report section are provided below. For Level 0 (Control group), a non-personalised dietary advice leaflet was provided at baseline and at month 3 of the intervention. At month 6 of the intervention (end of the study) all participants from Level 0, 1 and 2 received a full report, including all sections.

Table S3. Section include in the participants feedback report by intervention arm.Section included in the report Level 1

(dietary advice)Level 2

(dietary + phenotypic advice)

Level 3(dietary +

phenotypic + genetic advice)

Section 1. A message from your nutritionist (Fig 1.)

✓ ✓ ✓

Section 2. How your diet compares to recommendations (Fig 2.)

✓ ✓ ✓

Section 3. Your physical characteristics (Fig 3.)

✓ ✓ ✓

Section 4. Your Physical Activity Level (Fig 4.)

✓ ✓ ✓

Section 5. Your nutrient profile (Fig 5.) ✓ ✓ ✓Section 6. Your blood profile relating to nutrition (Fig 6.)

✓ ✓

Section 7. Your genetic profile relating to nutrition (Fig 7.)

✓

Section 8. Your Personalised Nutrition Advice (Fig 8.)

✓ ✓ ✓

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Figure 1. Example of ‘A message from the nutritionist’ (Section 1). Note: This section was a short paragraph of about 100 – 150 words max. The aims of the text were to personalise the report, to encourage the participant and to summarise the main message relating to improvement or progress since the last report.

Figure 2. Example of ‘How your diet compares to recommendations’ feedback (Section 2). Note: This section of the report showed the participants how their diet compares with the ‘Dietary Guidelines’. This was displayed to the participants in table format using coloured images to represent the number of portions of each food group they are consuming.

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Figure 3. Example of ‘Your Physical Characteristics’ feedback (Section 3)Note: Within this section of the report participants received information on their self-reported body measurements.

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Figure 4. Example of ‘Your Physical Activity level’ feedback (Section 4)Note: Within this section of the report participants received information on their self-reported physical activity level at work, during leisure and overall collected with the Beacke PA questionnaire (Section 5.a) and objectively measured PA level using accelerometer (Section 5.b).

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Figure 5. Example of ‘Your Nutrient Profile’ feedback (Section 5)Note: This section of the feedback report illustrated, graphically, how the participant’s intake of selected nutrients, vitamins and minerals compared with the IOM recommendations.

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Figure 6. Example of ‘Your Blood Profile’ feedback (Section 6)Note: This section of the feedback report illustrated, graphically, how the participant’s blood results for cholesterol, glucose, omega-3 and carotenoids compared with recommendations. Only participants randomised to Level 2 and Level 3 received these results during the intervention.

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Figure 7. Example of ‘Your Genetic Profile’ feedback (Section 7)Note: This section of the feedback report summarised the participant’s individual carriage of the 5 selected genetic variants (see Table S2). The statements “Yes” or “No” in the table reflected whether participant’s carried (or not) the risk variant of each genetic variant. Only participants randomised to Level 3 received these results during the intervention.

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Figure 7. Example of ‘Your Personalised Nutrition Advice’ feedback (Section 9)Note: This section of the feedback report is split into two sections 1) body weight and physical activity recommendations, and 2) dietary goals. For the first section, the nutritionist provided the participant with appropriate weight and physical activity recommendations. In the second section of the feedback report, the nutritionist identified the priority genes/nutrients/blood –related markers on which advice to the individual participant was based. These top 3 targets were selected on the basis of risk, i.e. the top 3 targets were those that were furthest from the recommended intake or level for that individual participant. For participants randomised to L1, personalised advice was based on dietary intake only, for those randomised to Level 2, personalised advice was based on dietary intake and phenotypic characteristics, and for Level 3 participants dietary intake, phenotype and genotypic information were all used to derive the personalised advice.

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Example of advice provided to participants randomised to L0 “Control group”

Table S4. No-personalised dietary eating guidelines

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Eat a varied diet that is primarily plant-based including plenty of fruit, vegetables, wholegrain and fish. Limit your intake of red meat, salt, added sugar and energy-rich food products.

A varied diet may be the best way to maintain and improve your health and ensure an optimal nutrient intake.

Maintain a balance between energy intake and energy expenditure. Energy intake from food and drink and energy expenditure through

physical activity should be balanced to maintain body weight within a normal range.

For individuals who are overweight, it is recommended to increase your physical activity alongside an energy-reduced diet.

Eat at least 5 portions of fruit and vegetables every day. Eat a minimum of 5 portions of fruit (including berries) and vegetables

every day. This corresponds to at least 400 g as one portion is about 80 g (this is about the size of your fist or a medium sized apple).

Eat a variety of fruit and vegetables of different colours (including red, green, yellow, white, violet and orange).

Fresh, tinned, frozen and cooked vegetables and fruit (boiled, microwaved, baked or fried) can all be included as part of your recommended intake. Choose products with no added sugar.

Dried fruit can be included as part of your recommended intake, but the portion size should be halved. One glass of juice or smoothie, as well as legumes, can be included in the five portions per day. However, no matter how much you drink or eat of these items, they will only count as one portion.

Potatoes are not included in your five portions of fruit and vegetables per day. However, potatoes can certainly be included in a varied diet. Nuts are not included in the five portions per day either, but a moderate consumption of nuts (approximately 20 g per day corresponding to 20 almonds) may be included in a varied diet. All nuts should be unsalted.

Eat wholegrain products every day. Eat at least 50 g wholegrain every day which can be obtained by eating

for example two slices of wholemeal bread, a small portion of wholegrain breakfast cereals or a portion of wholemeal pasta or brown rice. Breakfast cereals, porridge and crisp bread made with wholegrain are also good wholegrain sources.

Eat 2-3 portions of fish per week of which at least one should be oily.

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Eat fish 2-3 times per week. A portion of fish is approximately 150 g.

Both lean and oily fish can be included, but at least one portion should be oily.

Fish as a main meal can be replaced with fish as a sandwich topping. Three sandwich topping portions (50 g) of fish correspond to one main meal portion.

Oily fish include herring, sardines, halibut, mackerel, trout, salmon and tuna. Lean fish include cod and pollock.

Eat 3 low-fat dairy products per day.

Eat 3 portions of low-fat dairy products per day. One portion should be either one glass of milk (200 ml), a small pot of yoghurt (150 g) or small matchbox size piece/3 slices of cheddar-type cheese (30 g).

Low-fat dairy products are healthier. The consumption of dairy products containing high levels of saturated fat, such as full-cream milk, cream, fatty cheese and butter, should be limited.

Eat lean meat and lean meat products and limit your intake of red meat and processed meat.

Lean meat and lean meat products are important for most people to ensure an adequate consumption of a number of nutrients. Moderate consumption of lean meat can therefore be usefully included in the total diet.

Limit your consumption of red meat (beef, pork, lamb and goat) to 500 g per week. This corresponds to approximately 3 main meal portions per week, as one main meal portion is about 150 g (e.g. one chop or one burger). Note that cold cuts also are included in the 500 g.

Limit your consumption of processed meat products that are smoked, salted or preserved with nitrate or nitrite, for example sausages, ham, salami, bacon and Parma ham.

Use oil or soft margarine.

Include cooking oils or soft margarine in your diet. Choose cooking oils and margarine with high content of

unsaturated fatty acids, such as plant oils (e.g. rapeseed, sunflower, olive, soya, and walnut oil), and low content of saturated fatty acids (e.g. palm oil and animal fat).

Limit your use of butter and butter-margarine blends because these have high content of saturated fatty acids and low content of polyunsaturated fatty acids. Butter and animal fats also contain trans fatty acids and cholesterol.

Although you should limit your consumption of foods with a high energy content, cooking oils and soft or liquid margarines also contain polyunsaturated fatty acids and fat soluble vitamins. They should therefore be included in your diet.

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Water is the recommended drink.

Water should make up a large part of your fluid intake. If alcohol is consumed, limit the intake to no more than two

drinks a day for men and one drink a day for women. One portion is 1 glass of wine or 1 bottle of beer or 1 small glass of spirits.

Limit your intake of energy rich food products.

Limit your consumption of foods with a high energy content. That includes food with a lot of fat, oil and added sugar such as sweets, snacks, pastries, candies, and fast food.

These food products can be enjoyed occasionally and in small amounts (e.g. a handful). They are not meant for hunger, but for pleasure.

Limit your intake of salt.

Limit your intake of salt (sodium chloride) to a maximum of 6 g per day (which corresponds to 2.4 g of sodium). This equates to around 1 teaspoon of salt, and remember most of this is already included in foods.

Processed food products and ready meals often have a high salt content, whereas raw ingredients contain far less salt.

Try not to add salt when preparing, cooking or eating food. Use other flavourings such as herbs and salt-free spices instead of salt.

Be physically active for at least 30 minutes per day.

You should aim to be at least moderately physically active. You can achieve this simply by walking briskly for at least 30 minutes every day. This can be incorporated into occupational, transport, household, or leisure activities.

As your fitness improves, aim for 60 minutes or more of moderate physical activity, or for 30 minutes or more of vigorous, physical activity every day.

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Appendix III: Nutrient Gradations and cut-off point for deriving personalised advice

Protein (g/kgBW)<0.52 >0.52 & <0.66 >0.66 & <2.4 >2.4

Carbohydrate(% of total energy)

<40% >40 & <45% 45-65% >65 & <70% >70%

Total Fat(% of total energy)

<15% >15 & <20% 20-35% >35 & <40% >40%

Monounsaturated(% of total energy)

<10% >10 & <15% 15-20% >20 & <30% >30%

Polyunsaturated(% of total energy)

<5% >5 & <6% 6-11% >11 & <12% >12%

Saturated Fat(% of total energy)

<10% >10 &<15% >15

Salt (g)

18-50yrs<3.75 >3.75 & <5.75 >5.75

51-70yrs <3.25 >3.25 & <5.75 >5.75

>70yrs<3 >3 & <5.75 >5.75

Omega-3(% of total energy)

<0.2 >0.2 & <0.6 >0.6

Fibre (g)Males

18-50yrs<28 >28 & <38 >38

>50yrs <20 >20 & <30 >30

Fibre (g)Females

18-50yrs<15 >15 & <25 >25

>50yrs <14 >14 & <21 >21

Calcium (mg)Males

18-70yrs <600 >600 & <800 >800 & <2500 >2500

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>70yrs <800 >800 & <1000 >1000 & <2500 >2500

Calcium (mg)Females

18-50yrs <600 >600 & <800 >800 & <2500 >2500

>50yrs<800 >800 & <1000 >1000 & <2500 >2500

Iron(mg)Males >18yrs <4 >4 & <6 >6 & <45 >45

Iron(mg)Females

18-50yrs<3.15 >3.15 & <8.1 >8.1 & <45 >45

>50yrs <3.5 >3.5 & <5 >5 & <45 >45

Vitamin A (µg)Males

<350 >350 & <625 >625 & <3000 >3000

Vitamin A (µg)Females

<300 >300 & <500 >500& <3000 >3000

Folate (µg) <240 >240 & <320 >320 & <1000 >1000

Thiamin (mg)Males

<0.8 >0.8 & <1.0 >1.0

Thiamin (mg)Females

<0.7 >0.7 & <0.9 >0.9

Riboflavin (mg)Males

<0.9 >0.9 & <1.1 >1.1

Riboflavin (mg)Females

<0.7 >0.7 & <0.9 >0.9

Vitamin B12 (µg) <1.6 >1.6 & <2.0 >2.0

Vitamin C (mg)Males

<60 >60 & <75 >75 & <2000 >2000

Vitamin C (mg)Females

<45 >45 & <60 >60 & <2000 >2000

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