Universal nutrime pellet concept

OVERVIEW

How genetic variants can disrupt protective systems in our body

MICRONUTRIENTS Genes fulfill a number of protective roles in the body OXIDATIVE STRESS

GPX1 SOD2

TNFa

AGRESSIVE IMMUNE SYSTEM

IL1A

MTHFR

REGULATE HOMOCYSTEINE

MTRR

GSTM1

REMOVE TOXINS FROM BODY

GSTT1

MICRONUTRIENTS

Genetic variants disrupt protective effect OXIDATIVE STRESS

GPX1 SOD2

TNFa

AGRESSIVE IMMUNE SYSTEM

IL1A

MTHFR

REGULATE HOMOCYSTEINE

MTRR

GSTM1

REMOVE TOXINS FROM BODY

GSTT1

OVERVIEW

How micronutrients can recover some of the lost function

MICRONUTRIENTS

Certain micronutrients can recover lost function OXIDATIVE STRESS

GPX1 SOD2

Vit E Vit C

Vit A ALA

TNFa

AGRESSIVE IMMUNE SYSTEM

IL1A

MTHFR

REGULATE HOMOCYSTEINE

MTRR

GSTM1

REMOVE TOXINS FROM BODY

GSTT1

Folic acid Vit B12

Vit B6

Calcium Iron

Selenium

MICRONUTRIENTS The concept: Test 50+ genes and determine the requirement of 22 micronutrients Bone Health Osteoporosis (Calcium, Vit D, Magnesium)

Eye Health Macular Degeneration (Antioxidants)

Joint Health Rheumatoid Arthritis (Omega3)

Metabolic Health Iron overload disorder (Iron)

Heart health Cholesterol (Omega3) Homocysteine (Folic acid, B-Vitamins)

Food intolerances Lactose Intolerance (calcium) Coeliac disease (Multivitamin)

Cognitive Health Alzheimers disease (Antioxidants)

Detoxification Heavy metals (Calcium, Selenium, Iron)

OVERVIEW

One example of how genetics influence micronutrient requirement

OXIDATIVE STRESS Free radicals damage cells

5% of oxygen SOD2

Superoxide

Mitochondria

SOD3

SOD1

HydrogenPeroxide

5% of oxygen becomes Superoxide, a free radical. 3 genes protect the cell in various locations from these damaging chain reactions

CYTOPLASM OF A CELL

OXIDATIVE STRESS Free radicals damage cells

5% of oxygen

Superoxide

Mitochondria

SOD3

SOD1

HydrogenPeroxide

27% of the population has a genetic variant in this gene, abolishing its protective effect.

CYTOPLASM OF A CELL

OXIDATIVE STRESS Free radicals damage cells

5% of oxygen

Superoxide

Mitochondria

SOD3

SOD1

HydrogenPeroxide

Superoxide damages cells and speeds up ageing and disease development.

CYTOPLASM OF A CELL

OXIDATIVE STRESS Free radicals

OXIDATIVE STRESS Free radicals damage cells

5% of oxygen

ALA

ALA Vit A

Superoxide

Vit A

ALA

Vit E SOD3

SOD1

HydrogenPeroxide

Vit C Vit A

Vit C

Mitochondria

Antioxidants can help neutralize free radicals before they do any harm if available in the right quantities.

CYTOPLASM OF A CELL

OXIDATIVE STRESS Free radicals damage cells

5% of oxygen

ALA

ALA Vit A

Superoxide

Vit A

ALA

Vit E SOD3

SOD1

HydrogenPeroxide

Vit C Vit A

Vit C

Mitochondria

The lost protection from superoxide can be recovered

CYTOPLASM OF A CELL

OXIDATIVE STRESS Free radicals damage cells

SOD2 If SOD2 is active, it protects the cells. Only normal amounts of antioxidants are required.

SOD2

Vit A Vit E

ALA Vit C

e.g.

100mg

OXIDATIVE STRESS Free radicals damage cells

SOD2

SOD2 If SOD2 is deactivated, the protective function is lost.

Higher antioxidant dosages are necessary.

SOD2

Vit A Vit E

ALA Vit C

e.g.

100mg

Vit A Vit E

e.g.

ALA 300mg

Vit C

OVERVIEW

Another example of how genetics influence micronutrient requirement

OXIDATIVE STRESS

GPX1 – a Selenoprotein neutralizing free radicals The GPX1 gene produces an enzyme, that protects from oxidative stress as well.

GPX1

GPX1 Free Radical

OXIDATIVE STRESS

GPX1 – a Selenoprotein neutralizing free radicals Selenium

GPX1

GPX1 is a selenoprotein, meaning it needs dietary selenium to work.

GPX1 Free Radical

OXIDATIVE STRESS

GPX1 – a Selenoprotein neutralizing free radicals GPX1

GPX1

Selenium

Free Radical

OXIDATIVE STRESS

GPX1 – a Selenoprotein neutralizing free radicals GPX1

GPX1

Selenium

OXIDATIVE STRESS

GPX1 – a Selenoprotein neutralizing free radicals GPX1

GPX1

Selenium

Neutralized

OXIDATIVE STRESS Selenium deficiency and GPX1 activity

Selenium deficient 30% activity As selenium is required for GPX1 activity, selenium deficiency causes low activity and low protection.

OXIDATIVE STRESS Selenium deficiency and GPX1 activity more selenium has Selenium deficientAdding been shown to increase

30% activity

GPX1 activity and hence protection from oxidative stress.

More Selenium 70% activity

OXIDATIVE STRESS Polymorphism in the GPX1 gene 7% of the population carries a genetic variant that makes the enzyme lass effective.

GPX1

GPX1 Free Radical

OXIDATIVE STRESS Polymorphism in the GPX1 gene

GPX1

GPX1

Selenium

Free Radical

OXIDATIVE STRESS Polymorphism in the GPX1 gene

GPX1

GPX1

Selenium

Weaker binding (e.g. 50% weaker)

OXIDATIVE STRESS Selenium deficiency and GPX1 activity

Selenium normal 35% activity Even with normal selenium levels, the overall activity is low in carriers of this genetic variant.

OXIDATIVE STRESS Selenium deficiency and GPX1 activity

Selenium normal 35% activity

Increasing selenium levels to above normal levels recovers the lacking function of the enzyme.

MORE Selenium 60% Activity

OXIDATIVE STRESS Free radicals

OXIDATIVE STRESS

Dosage based on genetics

GPX1 If the person carries a functional GPX1-Gene, he receives the normal RDA as recommended by official agencies to be applicable to everyone.

55 Âľg / Day

OXIDATIVE STRESS

Dosage based on genetics

GPX1

GPX1 If the gene carries the variant, we increase selenium levels to recover the lost activity.

55 µg / Day

96 µg / Day

OVERVIEW

An example how some nutrients can have no effect

OXIDATIVE STRESS Coenzyme Q10 needs to be activated

Q10

Q10 – No effect Coenzyme Q10 has no antioxidant effect by itself and yet it is one of the most expensive Micronutrient available.

OXIDATIVE STRESS Coenzyme Q10 needs to be activated

Q10

Q10 – No effect NQO1

Q10

Ubiquinol Oxidative protection

UBI

The NQO1 Gene first must produce an enzyme converting inactive Coenzyme Q10 to active Ubiquinol.

OXIDATIVE STRESS Coenzyme Q10 needs to be activated

Q10

Q10 – No effect NQO1

Q10 Ubiquinol is a very potent free radical inhibitor.

Ubiquinol Oxidative protection

UBI

UBI

Free radical

OXIDATIVE STRESS Free radicals

OXIDATIVE STRESS Coenzyme Q10

Q10

Q10 – No effect

NQO1

Q10

Due to a genetic variant 30% of the population have less activity and 4% have no activity of the NQO1 gene.

OXIDATIVE STRESS Coenzyme Q10

Q10

Q10 – No effect

NQO1

Q10

Free Radicals

Free radicals accumulate and damage the cells

OXIDATIVE STRESS Coenzyme Q10

Q10 – No effect

Q10

NQO1

Q10 The only way to recover this function is through ingesting active Ubiquinol as supplement.

UBI

UBI

Free UBI Radicals UBI UBI

UBI UBI

OXIDATIVE STRESS Coenzyme Q10

Q10 – No Effect

Q10

NQO1

Q10 Alternatively, other antioxidants in higher dosages can protect from free radicals.

E ALA C

C

Free Radicals E

E E C

ALA

ALA

OXIDATIVE STRESS Free radicals damage cells

NQO1

Q10

People with active NQO1 genes can use Coenzyme Q10 as antioxidative support

OXIDATIVE STRESS Free radicals damage cells

NQO1

NQO1 People with inactive NQO1 genes require Ubiquinol and/or other antioxidants to recover lost function

E

Q10 Q10

C

UBI ALA

OVERVIEW

One example where micronutrient supply through food might be too low

LACTOSE INTOLERANCE Genetics of Lactose intolerance

GENETICS Normal process in a baby Lactose can not be absorbed in the intestine Glucose

GLU LactASE enzyme splits lactose into smaller absorbable sugars Galactose

GLU

Babies produce this enzyme throughout the first years of their life to be able to digest milk

Lactase-Gene

GENETICS Normal process in an adult

GLU Lactose is food source for bacteria

GLU Evolutionarily, Adult (cavemen) had no access to milk, needed no enzyme and so production was gradually swithed off with increasing age.

Gene is deactivated with increasing age

Age++

Lactase-Gene

GENETICS Lactose intolerance – a normal process in adults

Lactase-Enzyme

The normal process is a gradual decrease in enzyme production

1 in 6 caucasians: Lactase enzyme is deactivated

Light symptoms Intermediate Symptoms

Lactose intolerance

Severe symptoms 10

20

30

40 50 Age (Years)

60

70

GENETICS Genetic variant causing Lactose TOLERANCE in adulthood

GLU

GLU

SNP Genetic variant disrupts „Age-Deactivation“

Age++

Lactase-Gene

GENETIK Lactose intolerance 5 of 6 caucasians: Lactase enzyme is produced throughout life

Lactase-Enzyme

1 in 6 caucasians: Lactase enzyme is deactivated

Light symptoms Intermediate Symptoms

Lactose intolerance

Severe symptoms 10

20

30

40 50 Age (Years)

60

70

GENETICS How did lactose intolerance arise?

Genetic variation arose in one person in northern Europe 10 000 years ago

GENETICS How did lactose intolerance arise?

Famine caused many people to die

GENETICS How did lactose intolerance arise?

Milk as additional source of food = Survival & procreation Advantage 80% of europeans today carry this one genetic variant that arose back then.

GENETICS How did lactose intolerance arise?

Lactose tolerance

Lactose tolerance is hence only a european phenomenon

Lactose INtolerance

GENETICS How did lactose intolerance arise?

Lactose tolerance

Lactose tolerance was spread through colonization and emigration from europe

Lactose INtolerance

GENETICS Consequence for micronutrient dosing

Age++

Lactase-Gene

LACTOSE TOLERANT

= 600mg calcium through food

Lactose tolerant individuals thend to have a higher calcium intake and cover much of their requirement through their diet.

GENETICS Consequence for micronutrient dosing

Age++

Lactase-Gene

LACTOSE TOLERANT

= 600mg calcium through food

Ca Ca

200mg/day

Calcium RDA 800mg/day Since the RDA applicable to everyone is 800mg, some people dont get quite enough from their nutrition and would benefit from supplementation.

= 800mg calcium/day

GENETICS Consequence for micronutrient dosing

Age++

Age++

Lactase-Gene

LACTOSE INTOLERANT

LACTOSE TOLERANT

= 600mg calcium through food

People with a genetic variant in the LCT gene eat no milk products and ingest significantly less calcium through their diet.

Calcium RDA 800mg/day

Ca Ca

Lactase-Gene

200mg/day

= 800mg calcium/day

= 30mg calcium through food

GENETICS Consequence for micronutrient dosing

Age++

Age++

Lactase-Gene

LACTOSE INTOLERANT

LACTOSE TOLERANT

= 600mg calcium through food

Calcium RDA 800mg/day

= 30mg calcium through food

Ca

Ca

Ca Ca

Lactase-Gene

200mg/day

A higher supplementation is required to reach the RDA.

770mg/day

Ca Ca

Ca

= 800mg calcium/day

Ca Ca

OVERVIEW

One example where requirement of micronutrient might be higher than normal

GENETICS

Next question: Is the RDA enough?

Osteoporosis – brittle bones The question is, is the RDA enough or might some people need even more calcium?

OSTEOPOROSIS

Bone mineral density

Bone density and age

Bone density tends to increase up to the age of 30 and then gradually decrease with age.

Normal process: Bone density decreases slightly with age

Osteopenia Osteoporosis 10

20

30

40 50 Age (Years)

60

70

OSTEOPOROSIS Bone density and age

Bone mineral density

Normal process: Bone density decreases slightly with age Genetic variants: Bone density is lost rapidly

Osteopenia Osteoporosis 10

20

30

Genetic variants increase bone mineral density loss with increasing age.

Prevention is required.

40 50 Age (Years)

60

70

OSTEOPOROSIS Bone density and age

Bone mineral density

PREVENTION

Osteopenia Osteoporosis 10

20

30

As with saving money, prevention must be started as early as possible to be effective.

40 50 Age (Years)

60

70

OSTEOPOROSIS

Bone mineral density

Bone density and age

PREVENTION Osteopenia Osteoporosis 10

20

30

As with saving money, prevention must be started as early as possible to be effective.

40 50 Age (Years)

60

70

OSTEOPOROSIS

Bone mineral density

Bone density and age

Osteopenia Osteoporosis 10

20

30

PREVENTION As with saving money, prevention must be started as early as possible to be effective.

40 50 Age (Years)

60

70

OSTEOPOROSIS What dose of calcium is optimal?

1500 mg

The European food safety agency (EFSA) states the RDA of calcium to be 800mg. This applies to everyone.

800 mg

0 mg

RDA according to EFSA

OSTEOPOROSIS What dose of calcium is optimal?

1500 mg In scientific studies, 1200mg have been shown to be protective and reduce fracture risk.

1200 mg

800 mg

0 mg

Known to be effective in scientific studies

RDA according to EFSA

OSTEOPOROSIS What dose of calcium is optimal?

1500 mg 1200 mg

A person with ni genetic risk for osteoporosis should follow the RDA

average (low) risk

800 mg

800mg

0 mg

Known to be effective in scientific studies

RDA according to EFSA

OSTEOPOROSIS What dose of calcium is optimal?

1500 mg maximum (high) risk

1200 mg

1200mg

Known to be effective in scientific studies

A person with high osteoporosis risk should follow the dosages we know to be protective from osteoporosis.

average (low) risk

800 mg

800mg

0 mg

RDA according to EFSA

OVERVIEW

Another example where requirement of micronutrient might be higher than normal

GENETICS

Next question: Is the RDA enough?

Heavy metal detoxification

PHASE 2 DETOXIFICATION Enzymes remove heavy metals from body

Heavy metal

GSTM1

GSTT1 GSTP1

In Phase 2 detoxification, toxins are modified and removed from the body by GST Genes/enzymes

Removed through kidneys

Enzymatic modification

Neutralized

PHASE 2 DETOXIFICATION Enzymes remove heavy metals from body

Heavy metal

GSTM1

GSTT1 GSTP1

60% of the population have at least one of these genes deactivated through genetic variants. Toxins such as lead are not neutralized.

Not neutralized

Toxicity, Cancer etc.

PHASE 2 DETOXIFICATION Enzymes remove heavy metals from body

Heavy metal Calcium supplementation binds Lead

GSTM1

GSTT1 GSTP1

Ca

Ca Ca

Calcium has been shown to bind and remove lead from the body. Hence, calcium supplementation can recover lost function.

Removed through kidneys

Calcium binds toxins

OVERVIEW

How different cenetic factors collectively determine the optimal dose of a micronutrient

CALCIUM What influences the optimal calcium amount?

Recommended RDA

Lactose Intolerant

Osteoporosis Risk

Detoxification deficiency

NO +0 mg

NO +0 mg

NO +0 mg

YES +150 mg

YES NO +150 +0 mg mg

YES +100 mg

=1050mg

YES NO +150 +0 mg mg

YES +150 mg

YES NO +100 +0 mg mg

=950mg

YES +150 mg

YES +150 mg

YES +100 mg

=1200mg

=800mg

800 mg

Depending on genetic type, the calcium amount can be increased.

OVERVIEW

One example where one micronutrient has one effect in some and the opposite effect in other people

OMEGA 3 fatty acids An observation doctors frequently make

Person 1 with high cholesterol

Doctor recommends Omega3 supplements

Cholesterol improves

Most people assume fish oil capsules (omega 3) are beneficial for cholesterol und some are in fact right.

OMEGA 3 fatty acids An observation doctors frequently make

Person 1 with high cholesterol

Doctor recommends Omega3 supplements

Cholesterol improves

Person 2 with high cholesterol

Doctor recommends Omega3 supplements

Cholesterol becomes worse

What is the reason?

APOA1 (A/A)

APOA1 (G/G)

In some cases the opposite happens. HDL Cholesterol becomes worse.

OMEGA 3 fatty acids An observation doctors frequently make

Person 1 with high cholesterol

Doctor recommends Omega3 supplements

Cholesterol improves

Person 2 with high cholesterol

Doctor recommends Omega3 supplements

Cholesterol becomes worse

What is the reason?

APOA1 (A/A)

APOA1 (G/G)

This is caused by different genetic variations in the APOA1 gene.

OMEGA 3 fatty acids

What is the consequence?

Person 1 with high cholesterol APOA1 (A/A) Doctor recommends Omega3 supplements

Cholesterol improves

Person 2 with high cholesterol APOA1 (G/G) Doctor recommends Omega3 supplements

Doctor recommends Phytosterols!

Cholesterol improves Usind phytosterols instead of Omega-3 achieves the same cholesterol-improving effect.

Different micronutrients required to achieve same result

OMEGA 3 fatty acids

What daily dose is effective for improving HDL Cholesterol?

RDA: 250 mg The RDA for Omega-3 is only 250mg. Far below the dosages shown to be effective in scientific studies.

Amounts shown to be effective: 1000-2900mg

OMEGA 3 fatty acids

What daily dose is effective for improving HDL Cholesterol?

Daily dose 3000 mg

2900mg

Study 3

1500mg

Study 1

1000mg

Study 2

250mg

RDA according to EFSA

Ranges known to be effective among (also high risk) individuals

Range accepted to be sufficient for general population with average risk =RDA

0 mg

CHOLESTEROL Cholesterol is influenced by genes CETP

HDL

APOA5

Cholesterol OK

Genes that influence HDL cholesterol levels

In this case there is no genetic predisposition for bad HDL cholesterol levels

CHOLESTEROL Cholesterol is influenced by genes CETP

HDL

APOA5

Cholesterol OK

Genes that influence HDL cholesterol levels

APOA1

If APOA1 is of the beneficial variant, then Omega3 can be recommended at the low risk/RDA dose.

250mg OMEGA3

CHOLESTEROL Cholesterol is influenced by genes CETP

HDL

APOA5

Cholesterol OK

Genes that influence HDL cholesterol levels

APOA1

250mg OMEGA3

APOA1

250mg Phytoster.

If APOA1 reverses the effect of Omega3 fatty acids, the alternative, Phytosterols should be used instead.

CHOLESTEROL Cholesterol is influenced by genes CETP

HDL

APOA5

Cholesterol OK

Genes that influence HDL cholesterol levels CETP

HDL

APOA5

Cholesterol Too low

APOA1

250mg OMEGA3

APOA1

250mg Phytoster.

Certain genes cause bad HDL cholesterol levels. Stronger intervention neessary

CHOLESTEROL Cholesterol is influenced by genes CETP

HDL

APOA5

Cholesterol OK

Genes that influence HDL cholesterol levels CETP

HDL

APOA5

Cholesterol Too low

APOA1

250mg OMEGA3

APOA1

250mg Phytoster.

APOA1

1500mg OMEGA3

If APOA1 is beneficial, high doses of OMEGA 3 are recommended.

CHOLESTEROL Cholesterol is influenced by genes CETP

HDL

APOA5

Cholesterol OK

Genes that influence HDL cholesterol levels CETP

HDL

APOA5

Cholesterol Too low

If APOA1 is of the negative type, high doses of the alternative are given.

APOA1

250mg OMEGA3

APOA1

250mg Phytoster.

APOA1

1500mg OMEGA3

APOA1

1500mg Phytoster.

OVERVIEW

To summarize these few examples

SUMMARY Genes influence the requirement of Micronutrients Genes influence micronutrient effect Genes influence micronutrient dosage requirement By testing 50+ genes we can dose 22 micronutrients based on genes With 52 genes, there are 717 000 000 000 000 000 000 000 possible results Each micronutrient recipe is UNIQUE

OVERVIEW

How can one follow a specific micronutrient recipe?

CONCEPT OF NUTRIENTS How can you follow such variable recommendations?

Use standard supplements?

Dose always too high or too low

Individual gelatin capsules?

Very expensive to produce

Microtransporter method!

Easy to mix Easy to automate Every mix possible Cheap to produce

CONCEPT OF NUTRIENTS How can you follow such variable recommendations?

Vitamin C mix

Vitamin A mix

Zink mix

50% w/w

4% w/w

11% w/w

Molecule of filler Molecule of Vitamin C Molecule of Vitamin A Molecule of Zink

We simply add different amounts of different pellet types to a mixture, mix and then instruct to take one spoon per day

6g

38g

22g

Zink mix Mixture for one genetic profile

Vitamin A mix Vitamin C mix

Instructions: ingest 8g/day

Amounts differ depending on genetic profile

NUTRIME Pellets taken with spoon

OVERVIEW

What is the status of the product?

STATUS The status of the project

Patent pending (worldwide) Genetic test ready (50+genes) Microtransporters developed (23 different types) Label and instructions completed Product ready for launch

NUTRIME Materials so far

NUTRIME Materials so far

OVERVIEW

What can we offer?

OFFER What we can offer AMWAY

Genetic test to be performed for each individual (Ready to launch) Report booklet (digital or high quality print and post) describing results Highest level of certification (ISO15189, Austria for medical genetics) Personalized recipe for every person Pellet production using your high quality natural vitamins and minerals

Shipment logistics for every order

OVERVIEW

Important aspects to consider

TO CONSIDER

Are these tests not medical genetic tests?

One important aspect of nutrigenetic tests: The same test can be medical (if disease information is communicated), or a lifestyle test (if it only communicates nutrition advice).

LIFESTYLE

Makes you fat sensitive for obesity

PPARG

MEDICAL

Increases your risk of Diabetes by 38%

TO CONSIDER

Are these tests not medical genetic tests?

Lifestyle Genetic tests can be offered without limitations. Medical genetic tests are limited to qualified physicians.

LIFESTYLE

Lactose intolerance

MEDICAL

LCT

You need more calcium because of your genes

You are 95% likely to develop lactose intolerance during your lifetime

TO CONSIDER

Are these tests not medical genetic tests?

Another example:

LIFESTYLE

Osteoporosis

MEDICAL

LCT

You need more calcium because of your genes

Your risk of osteoporosis is 215% higher

TO CONSIDER

Are these tests not medical genetic tests?

Another example:

LIFESTYLE

Detoxification

MEDICAL

GSTM1

You need more antioxidants because of your genes

Your risk of cancer is 130% higher due to bad detoxification

TO CONSIDER

Are these tests not medical genetic tests?

Another example:

LIFESTYLE

OMEGA 3

MEDICAL

APOA1

You need Omega 3 for heart health

Omega 3 improves your HDL cholesterol (might be ok to communicate)

OVERVIEW

What about the science?

SCIENCE

Scientific background on the genes mentioned GENE: APOLIPOPROTEIN A1 (APOA1) Genetic Variant: rs670, G/A pos. -75 Promoter Disease risk: Effect of poly unsaturated fatty acid consumption on HDL (good) cholesterol Low risk genotypes: A/A or A/G show high HDL cholesterol levels High risk genotypes: G/G shows low HDL cholesterol levels The human Apolipoprotein A1 (APOA1) constitutes the major protein component of HDL (high-density lipoprotein; the socalled “good cholesterol”) in plasma. It promotes cholesterol efflux from tissues to the liver for excretion and is a cofactor for LCAT (lecithin cholesterol-acyltransferase) which is responsible for the formation of most plasma cholesteryl esters.

A relatively frequent promoter polymorphism (G/A pos. -75) in the human APOA1 gene modulates the transcription of the gene as shown in a series of in vitro experiments. Because low APOA1/HDL plasma levels constitute a well-known risk factor for coronary artery disease (CAD), scientists started to analyze and associate this polymorphism with plasma APOA1/HDL concentrations. Studies that examined this association reported contradictory results.

As a consequence, it was suggested that the inconsistencies between studies could be the result of interactions with environmental factors that modulate the effect of this polymorphism. Ordovas et al. (2002) studied this proposed interaction in a population-based sample (755 men and 822 women) from the Framingham Offspring Study and found out that polyunsaturated fatty acids (PUFA) intake significantly modulates the effect of the APOA1 G/A pos. -75 polymorphism.

In summary, in female carriers of the A-allele, higher PUFA intakes were associated with higher HDL-cholesterol concentrations, whereas the opposite effect was observed in G/G women. Their results illustrate the complexity of polymorphism-phenotype associations and underscore the importance of accounting for interactions between genes and environmental factors in population genetic studies. Subbiah MT (2007) wrote in a recent review on Nutrigenetics “From a dietary recommendation point of view, women with this mutation (A-allele carriers) should be counseled to consume higher levels of polyunsaturated fat.”

According to the current embodiment, a person carrying the genotype G/G for the rs670 genetic variant does not benefit from a high PUFA diet in terms of HDL cholesterol levels. PUFAS should not be a part of nutritional supplements in this case.

REFERENCES Angotti E. et al., 1994; PMID 8021234 Juo S.H. et al., 1999; PMID 10215547 Ordovas J.M. et al., 2002; PMID 11756058 Ordovas J.M., 2004; PMID 15070444 Subbiah M.T., 2007; PMID 17240315 Tuteja R. et al., 1992; PMID 1618307

SCIENCE

Scientific background on the genes mentioned

GENE: CADHERIN 13 (CDH13)

Genetic Variant: rs8055236 Disease risk: Cardiovascular disease Low risk genotypes: A/A (OR 1) High risk genotypes: A/G (OR1.91) and G/G (OR 2.23) The present variant describes a substitution from the genetic base A to the base G at the genetic location of the CDH 13 gene.

The CDH13 gene encodes a member of the cadherin superfamily. The encoded protein is localized to the surface of the cell membrane and is anchored by a GPI moiety, rather than by a transmembrane domain. The protein lacks the cytoplasmic domain characteristic of other cadherins, and so is not thought to be a cell-cell adhesion glycoprotein. This protein acts as a negative regulator of axon growth during neural differentiation. It also protects vascular endothelial cells from apoptosis due to oxidative stress, and is associated with resistance to atherosclerosis.

The Wellcome trust Case control Consortium demonstrated in 2007 on a study of 14,000 cases of seven common diseases and 3000 shared controls, that the risk of cardiovascular disease of a person is increased to an odds ratio of 1.91 in A/G heterozygotes and an odds ratio of 2.23 for homozygous mutants of this genetic variant when compared to the A/A wildtype (The welcome Trust Case Consortium, 2007). This genetic link between this genetic variant and cardiovascular disease was further demonstrated by Yan Y. in 2009 in the Framingham Heart Study Offspring Cohort.

Genetic evaluation: According to the current embodiment, a person carrying the genotypes A/G or G/G for the rs8055236 genetic variant are considered to be at an increased risk of cardiovascular disease REFERENCES The Wellcome Trust Case Control Consortium (2007): Nature. 2007 June 7; 447(7145): 661–678. Genome-wide association study of 14,000 cases of seven common diseases and 3,000 shared controls.

Yan Y (2009): BMC Proc. 2009 Dec 15;3 Suppl 7:S118. Evaluation of population impact of candidate polymorphisms for coronary heart disease in the Framingham Heart Study Offspring Cohort. Yan Y, Hu Y, North KE, Franceschini N, Lin D.

SCIENCE

Scientific background on the genes mentioned GENE: CORONARY HEART DISEASE, SUSCEPTIBILITY TO, 8 (CHDS8) Genetic Variant: rs1333049 Disease risk: coronary heart disease Low risk genotypes: G/G (OR 1) High risk genotypes: G/C and C/C Two of the most frequently and thoroughly studied genetic variants for coronary heart disease susceptibility are SNPs rs10757278 and rs1333049. These two SNPs are in strong linkage disequilibrium (LD, meaning that testing one completely predicts the states of the other genetic variant) and are practically “equivalent”. In view of this strong LD specific risk evaluation can be done by only genotyping rs1333049.

First identified in a study of the Wellcome Trust Case Control Consortium (2007; approx. 2.000 cases and 3.000 controls) the rs1333049 SNP was associated with CAD having an OR of 1,47 and 1,90 for hetero- and homozygotes respectively. Additional large studies could replicate this finding. A meta-analysis confirmed again the association of rs1333049 with CAD in 12.004 cases and 28.949 controls with an OR of 1,24 per allele. In the analysis ofSchunkert et al. (2008) there was no interaction concerning the risk of CAD between rs1333049 and numerous traditional risk factors, including history of MI; the population attributable risk (PAR) was 22 %. Furthermore Samani NJ. et al. (2007) demonstrated, that in their model the prediction of MI on the basis of the Framingham/PROCAM score was improved by using the rs1333049 genotyping information.

The association of this locus with CAD was also confirmed with the equivalent rs10757278 by Helgadottir A. et al. (2007) in a total of 4.587 cases and 12.767 controls all of European origin. The association of the G-allele of rs10757278 with CAD was highly significant resulting in an OR of 1,26-1,28 for heterozygotes and 1,64 for homozygotes according to an autosomaladditive model. This effect was even more pronounced in case of early-onset (men≤50 years, women≤60 years) myocardial infarction (MI) with an OR of 1,49 and 2,02 respectively.

According to the current embodiment, a person carrying the genotypes G/C or C/C for the rs1333049 genetic variant are considered to be at an increased risk of cardiovascular disease.

REFERENCES The Wellcome Trust Case Control Consortium (2007): Nature. 2007 June 7; 447(7145): 661–678. Genome-wide association study of 14,000 cases of seven common diseases and 3,000 shared controls.

Helgadottir A. et al., 2008; PMID 18176561 Helgadottir A. et al., 2007; PMID 17478679

SCIENCE

Scientific background on the genes mentioned GENE: APOLIPOPROTEIN A5 (APOA5) Genetic Variant: rs662799 Disease risk: Cardiovascular disease and hypertriglyceridemia Low risk genotypes: T/T (OR 1) High risk genotypes: T/C (OR 1.98) or C/C (OR1.98) Apoliporotein A-V (A5) is an important regulator of several lipoprotein fractions including plasma triglyceride (TG) and HDLcholesterol. The mature APOA5 protein is expressed in the liver only and secreted into the plasma. One of the most frequently studied polymorphism of the APOA5 gene is -1131 T/C (located in the promoter region). The genotype frequency of – 1131 T/C differs between populations: The MAF (Minor Allele Frequency) of the C-allele is ≈ 30 % in Japanese while only ≈ 12-15 % in Caucasians. The variant - 1131 C-allele is associated with significantly higher plasma triglyceride in multiple populations (Japan, USA, Germany, Hungary, UK etc.) and in several cases with significantly decreased HDL-cholesterol concentrations. Both are basic characteristics of the Metabolic Syndrome (MS) and are established risk factors for Cardiovascular Disease (CVD).

Yamada et al. (2007) reported in 1788 subjects that the -1131 C- allele was significantly associated (OR=1,57) with Metabolic Syndrome according to a dominant model, plasma triglyceride were significantly higher and HDL lower than in T/T carriers. In a study performed by Maász et al. (2007) the 1131 C- allele – was again significantly associated with the Metabolic Syndrome, with higher plasma triglyceride and lower HDL- concentrations. Most of the studies however yielded only significant associations of –1131 T/C variant with higher triglyceride levels and in several fewer cases with lower HDL. Szalai et al. (2004) investigated the possible association of –1131 T/C polymorphism with Coronary Artery Disease (CAD) in 308 patients referred to coronary bypass surgery and in 310 controls. The prevalence of –1131 C-allele was significantly higher in CAD patients, 10,9 % against 5,7 % (p < 0,001). After multiple adjustment: OR=1,98 for developing CAD in risk-allele carriers.

Vaessen et al. (2006) investigated the relationship between APOA5/Triglyceride and CAD in a nested case-control, the Epic-Norfolk Population Study (n= 1034 cases / 2031 controls). The minor –1131 C-allele was significantly associated with higher triglyceride concentrations and its frequency was higher in CAD cases.

According to the current embodiment, a person carrying the genotypes T/C or C/C for the rs662799 genetic variant are considered to be at an increased risk of cardiovascular disease.

REFERENCES Yamada Y. et al., 2007; PMID 16806226 Maász A. et al., 2007; PMID 17922054 Szalai C. et al., 2004; PMID 15177130 Vaessen S.F. et al., 2006; PMID 16769999

SCIENCE

Scientific background on the genes mentioned GENE: QUINONE ACCEPTOR OXIDOREDUCTASE 1 (NQO1)

Genetic variant: rs1800566, Pro187Ser Disease risk: Reduced conversion of Coenzyme Q10 to ubiquinol and increased oxidative stress High enzyme activity genotypes: C/C Low enzyme activity genotypes: C/T or T/T NAD(P)H dehydrogenase [quinone] 1 is an enzyme that in humans is encoded by the NQO1 gene. This gene is a member of the NAD(P)H dehydrogenase (quinone) family and encodes a cytoplasmic 2-electron reductase. This FAD-binding protein forms homodimers and reduces quinones to hydroquinones.

Coenzyme Q10 is an oil-soluble, vitamin-like substance and present in most eukaryotic cells, primarily in the mitochondria. It is a component of the electron transport chain and participates in aerobic cellular respiration, generating energy in the form of ATP. Ninety-five percent of the human body’s energy is generated this way. Therefore, those organs with the highest energy requirements—such as the heart, liver and kidney—have the highest CoQ10 concentrations. There are three redox states of coenzyme Q10: fully oxidized (ubiquinone), semiquinone (ubisemiquinone), and fully reduced (ubiquinol). The capacity of this molecule to exist in a completely oxidized form and a completely reduced form enables it to perform its functions in the electron transport chain and as an antioxidant respectively.

In order for Coenzyme Q10 to act as an antioxidant, the molecule needs to be converted to Ubiquinol and this transition is performed by the NQO1 enzyme. The genetic variant rs1800566 produces a completely inactive enzyme and the conversion of Coenzyme Q10 is significantly slowed. Therefore supplementation of Coenzyme Q10 has no beneficial health effect in carriers of the homozygous polymorphism.

According to the current embodiment, a person carrying the genotype C/T is considered to be an intermediate metabolizer of Coenzyme Q10 and a person carrying the T/T genotype is considered to be a poor metabolizer of Coenzyme Q10.

REFERENCES PMID: 9271353 PMID: 16551570 PMID: 10208650 Aberg F, EL Appelkvist, G Dallner, and L Ernster. Distribution and redox state of ubiquinones in rat and human tissues. Arch Biochem Biophys 295 (2): 230–4, 1992.

Alleva R, M Tomasetti, M Battino, et al. The roles of coenzyme Q10 and vitamin E on the peroxidation of human low density lipoprotein subfractions. Proc Natl Acad Sci 92(20):9388-91, 1995.

Bentinger M, M Tekle, G Dallner. Coenzyme Q – biosynthesis and functions. Biochem Biophys Res Commun 396(1):74-79, 2010.

Bhagavan HN and RK Chopra. Coenzyme Q10: Absorption, tissue uptake, metabolism and pharmacokinetics. Free Radical Research 40 (5): 445–53, 2006.

OVERVIEW

And many more…

TOUR OF THE LAB

A short tour of our facilities

Novogenia reception area

DNA extraction laboratory

DNA extraction laboratory

storage rack for swabs currently being processed

DNA analysis laboratory

meeting room and video conferencing station

meeting room and video conferencing station

head office

Dr. Daniel Wallerstorfer Founder and CEO

Sandra at the recetion desk

Maria at the reception desk

Saskia bringing new samples for analysis

paper forms are photographed and digitalized

we write our own software

presentations and trainings held via Skype

Florian at the sample processing queue

Saskia and Michael planning processes at the board

Thomas sterilizing his hands

Daniel picking up swabs to be processed

Jenny at our DNA extraction robot #1

Saskia checking the validity of barcoded swabs

Florian preparing DNA extraction

swabs are scanned and registered in the system

one swab goes into one slot in a 96 well plate

the swabs are trimmed

a full plate of swabs is moved to the DNA extraction robot

192 swabs are extracted in 1,5h

DNA is the transferred into a DNA plate

DNA extraction is fully automated

we have 3 large DNA extraction robots

and 2 small DNA extractors

total extraction capacity: 100 000 swabs per month

we use high capacity 384 well plates

these can analyze 72 samples at once

mixing DNA with reagents is also automated

plates are cooled during preparation for analysis

Daniel preparing a liquid handling robot

Robots use filter tips to avoid contamination

Reagents are prepared in UV sterilized cabinets

Maria preparing the analysis process

the Viia7 is the most advanced DNA analyzer

the machine can analyze 384 genes in 1h

adding up to 720 samples per 10h day

we operate a loading robot for 24/7 operation

the robot removes completed plates and loads new ones

plates are barcode labeled and automatically recorded

total analysis capacity is 51 000 samples per month

every bay holds 45 plates or 3200 customers

our alternative DNA analyzer in ring-format

office area â&#x20AC;&#x201C; we are largely paper less

opened swabs are sprayed with DNA degrading chemicals