Bruce N. Ames

Children’s Hospital Oakland Research Institute

Professor, University of California, Berkeley

A DIET FOR HEALTH AND

LONGEVITY

18 November 2010

Chicago NHRI

~40 Essential Micronutrients

• Biotin

• Folic acid

• Niacin

• Pantothenate

• Riboflavin

• Thiamine

• VitA

• VitB6

• VitB12

• VitC

• VitD

• VitE

• VitK

• Calcium

• Chloride

• Chromium

• Cobalt

• Copper

• Iodide

• Iron

• Magnesium

• Manganese

• Molybdenum

• Phosphorus

• Potassium

• Selenium

• Sodium

• Zinc

• Linolenic acid/DHA [ -3]

• Linoleic acid [ -6]

• Isoleucine

• Leucine

• Lysine

• Methionine

• Phenylalanine

• Threonine

• Tryptophan

• Valine

• Histidine

• Choline

Micronutrient Undernutrition in Americans

Nutrient Population Group

% Ingesting

< EAR *

From Food

Iron Menstruating Women 16 %

Magnesium All 56 %

Zinc All 12 %

B6 Women > 70 years 49 %

Folate Adult Women 16 %

E All 93 %

C All 31 %

Minerals

Vitamins

Very low intake:vitamins D & K, calcium, potassium, omega-3

•USDA What we Eat in America (NHANES 2001-2002) Sept. 2005

Micronuclei in: RNA positive erythrocytes

RNA negative erythrocytes

Folic AcidFolinic

Acid

TIME (DAYS)

1 year

preRx

Normal

range

Mic

ron

ucle

i p

er

1000

cells

20

50 100 150 200 250 300 350

0

30

40

80

130

Everson RB, Wehr CM, Erexson GL, and MacGregor JT. (1988) J Natl Cancer Inst 80:525-9.

dUMP dTMP

Methionine

SHMT

B6

Serine

MTHFR

Homocysteine

(polymorphism)

TS

MS

CH2=THF

B12CH3-THF

Dose-response on micronuclei induction in cultured lymphocytes

Acute exposure to X-rays vs. Folic Acid deficiency

Fenech 2003, Nutrition Research Reviews

Each of the six dependent variables (that were analyzed by nonlinear regression in former figures) were transformed to Z

scores and modeled as a quadratic function of the ln-liver nonheme iron as the independent variable. The equation for the

RCR ratio's Z score was obtained from inverted RCR values (1/RCR) so that normal rats had the lower instead of the higher

values. For presentation purposes each model line was obtained from 9 values of liver iron. All statistics were performed as in

materials and methods.

Analysis of nonlinear regression models: comparison of an overall model

and individual models of Z-transformed values vs. ln- nonheme liver iron

Over all

DCF-PMNs

DCF-Lymph

Rh123-PMNs

Rh123-Lymph

mtDNA damage

1/RCR

-1.5

-1

-0.5

0

0.5

1

1.5

2

2.5

3

-1.5 -1 -0.5 0 0.5 1 1.5 2 2.5

LN nonheme Fe (µmol/g wet liver)

Z s

co

re

normal

20

40

60

80

100

120

140

Control ZnAD ZnDF

DC

F F

luo

res

ce

nc

e

Inte

ns

ity (

RF

U)

Zinc Deficiency Induces Increased

Oxidative Stress in C6 Glioma Cells

*

Zinc Deficiency Induces Fapy Glycosylase (Fpg)-sensitive

Single Strand Breaks in Human Lung Fibroblasts

0

40

80

120

160

200

Control ZnAD ZnDF

Co

me

t S

co

re

Control (+Fpg) ZnAD (+Fpg) ZnDF (+Fpg)

*

Mitochondrial

Matrix

Cellular Cytoplasm

Mitochondrial Outer Membrane

Inner Membrane

Intermembrane

Space

V

I

IIIII

IV

CytCCoQ

H+

Pyruvate

Dehydrogenase

complex

Citrate

Synthase

-Ketoglutarate

Dehydrogenase

Complex

Fumarate

L-Malate

Oxaloacetate

Acetyl-Co-A Citrate Isocitrate

-Ketoglutarate

Succinyl-Co-A

Succinate

FADFADH2

CITRIC

ACID

CYCLE

H2OO2 ADP

ATP

ATP

NADH

NADH

NADH

NAD+

NADHNADH

H+

H+

H+ H+

H+H+H+

H+

H+H+

H+ H+H+

H+

H+

H+

H+ H+H+

H+

H+

H+

9

Magnesium Deficiency Induces DNA-Protein Cross-links

Calcium Deficiency Vitamin B12 Deficiency Fenech: chromosome breaks Fenech: Chromosome breaks

Lipkin: colon cancer mice

Folate Deficiency Selenium DeficiencyMacGregor/Ames/Fenech: chromosome Rao: DNA damage

breaks mice/humans Combs/Trumbo: Cancer humans

Willett: epi colon cancer humans

Vitamin D Deficiency Omega-3 FA DeficiencyHolick: epi many types of cancer Denkins: Cancer

Magnesium Deficiency Niacin DeficiencyBell: chromosome breaks humans Kirkland/Depeint: DNA damage

Larsson: epi colorectal cancer humans

Zinc Deficiency Choline DeficiencyFong: esophageal cancer humans/rodents da Costa: DNA damage in humans

Potassium DeficiencyChang: Cardiovascular Disease

Proc. Natl. Acad. Sci. USA

Vol. 103, pp. 17589-17594, November 2006

Low micronutrient intake may accelerate the degenerative diseases

of aging through allocation of scarce micronutrients by triage

Bruce N. Ames

Children’s Hospital of Oakland Research Institute, Nutrition and Metabolism Center,

5700 Martin Luther King Jr. Way, Oakland, CA 94609

• Most of the world’s population has inadequate intake of one or more micronutrients.

• Triage theory posits as a results of recurrent shortages of micronutrients during evolution, natural

selected developed a metabolic rebalancing response to shortage.

• The rebalancing favors micronutrient-dependent protein needed for short term survival while

those only required for long-term health are starved.

• This impairment results in insidious damage (e.g. increased DNA damage) that, over time, leads

to the acceleration of age-associated diseases (e.g. increased cancer).

40 Essential Micronutrients

• Biotin

• Folic acid

• Niacin

• Pantothenate

• Riboflavin

• Thiamine

• Vitamin A

• Vitamin B6

• Vitamin B12

• Vitamin C

• Vitamin D

• Vitamin E

• Vitamin K

• Calcium

• Chromium

• Cobalt

• Copper

• Fluoride

• Iodine

• Iron

• Magnesium

• Manganese

• Molybdenum

• Phosphorus

• Potassium

• Selenium

• Sodium

• Zinc

• Linolenic acid/DHA [ -3]

• Linoleic acid [ -6]

• Isoleucine

• Leucine

• Lysine

• Methionine

• Phenylalanine

• Threonine

• Tryptophan

• Valine

• Histidine

• Choline

16 Vitamin K Dependent Proteins

F2 (Prothrombin)

F7

F9

F10

(Anticoagulant protein C)

Osteocalcin

Gas 6 protein

Matrix Gla protein

TGFBI

Periostin

(Anticoagulation Protein Z)

Coagulation Factors Other Proteins

glutamyl-carboxylase, vitK quinone reductase

and vitK epoxide reductase)

5 Lethal KO 6 nonlethal KO

Non-lethal

VKD-

protein

Function Mouse

Knockout

Phenotypes

Human

Mutants

Anticoagulant

Therapy

Modest

VitK

deficiency

Osteocalcin

Matrix Gla

Protein

(Mgp)

Bone struct.

Glucose

homeostasis

Negative

regulator of

vascular

calcification

Fragile bones

Insulin

resistance

Arterial

calcification

BMD loss

(SNPs)

Abnormal soft

tissue

calcification

(Keutel

Syndrome;

SNPS)

Bone health

(men/children/rats)

Arterial

calcification

(humans/rats)

Bone health

Insulin

resistance

Arterial

calcification

Four Causes of Functional Deficiency of VKD-

Proteins in Humans

McCann & Ames (2009) Vitamin K, an example of triage theory: is micronutrient

inadequacy linked to diseases of aging? Am J Clin Nutr 90,889-907.

Natto

Selenoprotein genetic loss Se dietary loss

Rodent Human Human

Gpx1 Knockdown: UV-induced

micronuclei

Gpx1 SNP, heterozygosity loss,

hypermethylation: Various cancers

DNA damage/Cancer

Gpx2 KO: UV-induced cancer

Gpx1 Heterozyg: Heart Abnormalities Heart disease: hypertension,

mortality;

Keshan’s cardiomyopathy

(severe def)Gpx3 SNP: Stroke & thrombosis

Dio2 SNP: Hypertension (?)

Gpx1 KO: Viral induced myocarditis

Sensitivity to virus infection

Gpx2 KO: Airway inflammation

McCann, JC & Ames, BN. An examination of selenoprotein biology from the perspective of the triage theory: Why modest selenium deficiency may increase risk of diseases of aging. October, 2010 (to be submitted)

Modest Selenium deficiency, genetic impairment of nonessential

selenoproteins, and diseases of aging

An overview of evidence for a causal relationship

between iron deficiency

during development and cognitive or behavioral

function in children

Joyce C McCann and Bruce N Ames

(2007) AJCN

Is docosahexaenoic acid, an n-3 long-chain

polyunsaturated fatty acid, required for development

of normal brain function? An overview of evidence

from cognitive and behavioral tests in humans and

animals

Joyce C McCann and Bruce N Ames

American Journal of Clinical Nutrition (2005)

82:281-95

Is there convincing biological or behavioral evidence linking

Vitamin D deficiency to brain dysfunction?

Joyce C McCann and Bruce N Ames

Faseb J 22, 982-1001 (2008)

Neurons Contain The Enzyme That Activates Vitamin D

Eyles, DW et al (2005) J Chem Neuroanat 29, 21-30

Calcitriol Target Genes in the Brain

Gene products whose expression in the brain or brain cells has been reported to be affected by calcitriol

Neurotrophins and other growth factors:

NGF; NT-3 & NT-4/5; GDNF; TGF-β2

Calcium-binding proteins:

Calbindin D28K, parvalbumin, calretinin;

Protein sub-units for L-Type Voltage Sensitive Ca++Channels (L-type VSCCs);

Transcription factors or enzymes involved in signal transduction pathways:

N-myc, c-myc, protein kinase C family (PKC);

Other enzymes:

Choline acetyltransferase, responsible for synthesis of the neurotransmitter acetylcholine;

γ-Glutamyltranspeptidase, involved in recycling of the reactive oxygen species scavenger glutathione);

Hormones:

[Oxytocin, the “trust hormone”]

Biochemical or cellular brain functions in which calcitriol target gene products are involved

Synaptogenesis (formation of synaptic connections); Synaptic plasticity (e.g., memory formation); Calcium

signaling and homeostasis; Neurotransmission and neurotransmitter synthesis; Survival and differentiation of

dopaminergic and other neurons; Control of toxic free radicals;

Behavior affected by target gene product dysfunction

Learning and memory; Motor control; Maternal or social behavior; Aging (neuronal density);

Immune Risk Phenotype of Aging

Lower CD4 and CD8 T-lymphocytes

Increase in anergic effector (CD8+CD28-) T-cells

Low lymphoproliferative response

Decline in antigen-presenting cells

Decreased expression of co-stimulatory molecules

Decline in IL-12 production and Th1 response

Immune Risk Phenotype of Aging

Lower CD4 and CD8 T-lymphocytes

Def: vit A., zinc, folate

Increase in anergic effector (CD8+CD28-) T-cells

Def: tryptophan, zinc,

Low lymphoproliferative response

Def: vit C, vit E, zinc. Vit B6

Decline in antigen-presenting cells

Def: vit E

Decreased expression of co-stimulatory molecules

Def: vit E, tryptophan. zinc

Decline in IL-12 production and Th1 response

Def: vit B6, Vit E, zinc

Fenech 2007, Forum Nutr.

Healthy Non-smoking

Males

Healthy Non-smoking

Females

Variation in chromosomal DNA damage rates within and between age

groups measured as MNC frequency.

The Economist, December 13, 2003

Dr. Allen Spiegel, NIDDK/NIH

Top Sources of “Nutrition” for 20-30 year olds

NHANES III (1988-1994) Wakimoto P & Block G. J Gerontol A Biol Sci Med Sci, 2001

% total energy

1. Regular soft drinks 8.8

2. Pizza 5.1

3. Beer 3.9

4. Hamburgers, meat loaf 3.4

5. White bread 3.3

6. Cake, doughnuts, pastries 3.3

7. French fries, fried potatoes 3.0

8. Potato chips, corn chips, popcorn 2.7

9. Rice 2.6

10. Cheese or cheese spread 2.5

38.6%

Supplementation Strategies

Conventional

• cumbersome

• compliance

• incomplete

CHORI Bar

• convenient

• complete

1. Moderate micronutrient deficiencies are widespread.

2. Triage theory postulates that moderate micronutrient deficiencies

can lead to accelerated aging and age-related diseases.

3. RDAs and EARs should be re-examined on the basis of triage theory,

which would put nutrition on a firm foundation.

4. The optimum intake of micronutrients will vary among people

due to polymorphisms.

CONCLUSIONS

• Bring the world’s population up to optimal intake with

inexpensive micronutrients.

• Identify people who need increased intakes of micronutrients

due to polymorphisms.

GOALS

80

75

70

65

60

55

50

45

401900 1910 1920 1930 1940 1950 1960 1970 1980 1990

46.4

4950.1

53.654.5

56.3

5861.3 61.4

65.7 65.666.7

71.173.2

74.9

77.5

78.9

71.4

69.967.1

Life Expectancy of Men and Women at Birth

SOURCE: National Institute on Aging

New Yorker, June 6, 2005

“You’re fifty-seven years old.

I’d like to get that down a bit.”

Jung

Suh

Mark

Shigenaga

Ash

Lal

Michele

Mietus-Snyder

Nutrition & Metabolism Center

Scientists

Sandy

Calloway

Joyce

McCannBruce Ames

Swapna

Shenvi

David

Killilea

END