1
Nutrition Beyond Calories &
the Role Trees can play
Navin Sharma
Science Seminar – 25 April 2014
2
• How knowledge is changing and how this will lead to future nutrition
• Double burden of nutrition
• Maternal nutrition and evolution of epigenetics
• From single molecule to total calories – a case study of aging research
• What will be the roles of trees?
DNA controling you  environment controlling you 
maternal nutrition  Mind controlling you??
Theme of the Seminar
3
Exogenous plant MIR168a specifically targets mammalian LDLRAP1:
evidence of cross-kingdom regulation by microRNA. Zhang L et al. Cell
Res. 2012 Jan;22(1):107-26
• Discovered plant micoRNAs in the serum of mice and humans.
• The microRNAs possibly came from ingested rice.
• The microRNA MI168a binds to the mRNA of low-density lipoprotein
receptor adapter protein 1, inhibiting its translation.
• Decrease LDL removal from plasma
• Ingested plant microRNAs can regulate the expression of human genes.
Micro RNA from food can be found in blood stream
MicroRNAs (miRNAs) are a class of small noncoding RNAs that control gene expression by targeting mRNAs
and triggering either translation repression or RNA degradation
4
Royal jelly is a complex, protein-rich substance secreted from
glands on the heads of worker bees.
To be queen larva is fed royal jelly inside a compartment
called a queen cup.
The workers and queens are genetically identical.
The royal jelly diet makes the queen: develops ovaries and a
larger abdomen for egg laying,
The worker are sterile.
The royal jelly silences a key gene (Dnmt3), which codes for
an enzyme that silences a group of queen genes. When
Dnmt3 is turned "on," the queen genes are epigenetically
silenced, and the larvae develop into the default "worker"
variety.
To be Queen – need special diet
Science. 2008 Mar 28;319(5871):1827-30.
"Genetic Science Learning Center, University of Utah, http://learn.genetics.utah.edu."
5
Jerdon’s Jumping Ant
Florida Carpenter Ant
ANT GENOME SEQUENCING –
WAY TO UNDERSTAND LONGEVITY
• In 2010 – The Ant Genome sequenced
• Jerdon‟s Jumping Ant & Florida Carpenter Ant
• Very Different Social Structure
• In Jumping Ant – Queen replaceable
• New Queen Fed Special Diet – Lives 10- times Longer
Up-regulation of telomerase and sirtuin deacetylases in longer-lived H. saltator
reproductives, caste-specific expression of microRNAs and SMYD histone
methyltransferases, and differential regulation of genes implicated in neuronal
function and chemical communication
Science (2010) – 329: 1068-1071
6
Malnutrition, per capita calories, Prevalence of diabetes and
CVD
Malnutrition Calories
Diabetes CVD
7
Source: Global Health Risks (2009)
Nutritional Double Burden
8
• Famine exposure in first 2 trimesters lead to 80% higher prevalence of overweight
(p<0.0005)
• Famine exposure in last trimester or famine exposure in first 5 months of life
associated with 40% lower prevalence of overweight (p<0.005)
Obesity in Young Men after Famine Exposure in Utero and
early Infancy (Ravelli et al NEJM, 1976)
The Fetal Origins or “Barker” Hypothesis
• Pioneered by David Barker in the early 1990s
– Painstaking development of historical birth cohort studies in
England
– Poor nutrition of mother during pregnancy: 2-fold increase
in coronary heart disease from largest to smallest BW &
Impaired GT increased from 14% to 40%
• Impaired fetal growth causes adult disease
• “Thrifty phenotype” - impaired fetal growth permanently
changes the body‟s structure and physiology
– Adaptive for compromised nutrition
– Maladaptive for over-nutrition
• Potential mechanisms include blood pressure & glucose
tolerance
Am J Clin Nutr 2000 71: 1344S-1352S.
10
11
Yellow Mouse Agouti Mouse
Maternal Supplements
with
zinc
methionine
choline
folate
B12
•High risk
cancer, diabetes, obesity
•Reduced lifespan
•Lower risk of
cancer, diabetes, obesity
• Prolonged life
LTR Hypomethylated LTR Hypermethylated
Hypermethylation leads to increase in lifespan and decrease in metabolic disorders
Cooneyetal.(2002)JNutr.132:2393S
Mother’s nutrition and epigenetic changes
"Genetic Science Learning Center, University of Utah, http://learn.genetics.utah.edu."
NUTRITION & HEALTH IN „OMICS ERA: EPIGENETICS
• CpG- Cytosine Phosphate
• Can be methylated to generate 5-methyl
cytosine
• Spontaneously deaminates to form
Thymine
• Poorly recognized by DNA repair
mechanism
• Thus CG  TG mutation propagates
12
Changes in gene expression, cellular memory, reversible, no change in DNA structure
Homeostasis Broken Down with methylation
Disease : Outcome of interaction of DNA and Food components
Figure 4. Resveratrol induces overall hyperacetylation in HepG2 but not Hep3B or HuH7 cells.
Venturelli S, Berger A, Böcker A, Busch C, et al. (2013) Resveratrol as a Pan-HDAC Inhibitor Alters the Acetylation
Status of Jistone Proteins in Human-Derived Hepatoblastoma Cells. PLoS ONE 8(8): e73097.
doi:10.1371/journal.pone.0073097
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073097
15
Nutrient or diet Epigenetic mechanism
Embryonic development Folate DNA methylation, imprinting
Choline DNA methylation
Protein restriction
DNA methylation, histone
modifications
Alcohol DNA methylation
Stem cell Butyrate
Histone acetylation, DNA
methylation
Retinoic acid PRC
Aging Folate DNA methylation
Calorie restriction Histone acetylation
Immune function Folate DNA methylation
Cancer Methyl-deficient diet Histone modification, microRNA
Genistein DNA methylation, microRNA
(−)-Epigallocatechin-3-gallate DNA methylation, PRC
Curcumin microRNA
Obesity, insulin resistance High-fat diet DNA methylation, microRNA
Methyl-deficient diet DNA methylation
Curcumin Histone acetylation
Inflammation Resveratrol Histone acetylation
AdoMet Histone methylation
Methyl-deficient diet microRNA
Neurocognition Choline
DNA methylation, histone
methylation
16
Add life to age – clear evidence from foods and nutrition
HEALTHY AGEING
http://www.healthandage.com/html/min/eama/eama6/idx/docs/june_2006/TSAL_J_POWELL_Healthy_Ageing.pdf
Oxidative stress appears to be a major
determinant of life-span accessible to
nutritional intervention.
17
SMR-Specificmetabolicrate
Increase in SOD may act as a determinant in longevity
ANTIOXIDANTS AND AGEING
AmJC/inNutr1991;53:373S-9S
MLSP- Maximum lifespan potential
18
ANTIOXIDANTS AND LIFESPAN
Increased concentration of antioxidants help in longevity
AmJClinNutr1991;53:373S-9S
19
Sod 2 null mutants show decreased life span in Drosophila
Genetics165:2295–2299(December2003)
A Sod2 NULL MUTATION IN DROSOPHILA
Life-span and superoxide dismutase/catalase
mimetics
Small synthetic mimetics increases the
mean life-span of C elegans by 44 %,
and treatment of prematurely aging
worms resulted in normalization of their
life-span (a 67 % increase).
MelovSScience2000Sep1;289(5484):1567-9
21
SOD 1 expression is higher in Male ant when compared to Queen
SOD1 gene expression as measured
by Northern blots
Sod1 EXPRESSION IN ANTS
SOD1 activity assay levels in
activity units per milligram
Lasius niger (Ant)
Queens reach an
age of 28 years,
whereas workers
for 1-2 years and
males, a few weeks
3486–3489 PNAS March 9, 2004 vol. 101 no. 10
Oops!!
22
sod-2 mutant worms live significantly longer than wild-type worms
PLoSGenetics:February2009|Volume5|Issue2
Sod-2 DELETION IN CAENORHABDITIS
ELEGANS
23
EFFECT OF CR AND EXERCISE ON SURVIVAL
IN RATS
Adapted and modified from Holloszy 1995
ClinicalInterventionsinAging2007:2(4)537–543
Median lifespan is increased with Calorie restriction
24
Credit:UniversityofWisconsin-Madison/JeffMiller CALORIC RESTRICTION IN MONKEYS
Caloric restriction slows ageing in monkeys
Resveratrol (3,5,4‟-trihydroxystilbene) mimics the effects
of dietary restriction
• Hypothesized to activate Sir2, or SIRT1 in
mammals
• Alternatively, hypothesized to upregulate AMPK,
downregulate mTOR (Mammalian target of
rapamycin)
Baur et al. 2006 Nature
26
ENHANCING ENERGY EXPENDITURE AND FAT
METABOLISM
Red grape (resveratrol) compound shows promise
Industry looking for exercise mimetics now!
www.medindia.net/news/tripepperblend
27
Basic
Functional
PrescriptiveCarbohydrates
Fats
Proteins
Vitamins
Minerals
Water
Flavonoids
Omega 3& 6
Hunger
Satiety
Basic functions
Driven by Cultural
& Ethnic Preferences
Preventive
Performance
Maintenance
Specific needs
For a condition
Driven by
Aspirational
needs
IndulgenceFunction
Doctor
Age
Sustenance Enjoyment / Health Health & Wellbeing
Well-being/
appearance
Age
Add life to
years
Evolution of Nutrition
Deva Kumari, Yashwanth Radhakrishnan & Navin Sharma (2012): International J of Life Science & Pharma
Res. 2(3):174-184
Agriculture
Life
Sciences
Forestry
Nutrition
Actives
How to improve nutrition of people in Developing World
28
Epigenetics
Human Genome
• New ways of working
• Newer / different alliances and partnerships
• Local knowledge
Focus in-and out-of-RDA
Barley Max Odorless Soy RS in Wheat
Amaranth Moringa
Agroforestry – Complete Nutrition
Nutrition Pyramid
29Methionine and Nyacin rich fruits: Peach, Jackfruit, Tamarind, Dates
Synsepalum dulcificum, a West African berry better
known as "miracle fruit," changes the way foods taste.
Possible model for the taste-modifying activity of MCL. (A) MCL is held on hT1R2-
hT1R3 at neutral pH (Left) and activates it under acidic pH (Right).
Koizumi A et al. PNAS
2011;108:16819-16824
MCA= Miraculin
MCL is widely used in Japan, where dedicated cafés are serving low-cal food
with a miracle fruit on the side
31
Low GI Palm Sugar
32
Low GI Maple Sugar
Oil with High Oleic / High Stearic Acid
Margarines and spreads with cardio neutral saturated fatty acid – no chemical
hydrogenation & no trans fats
Enhanced functionality with higher heat stability for frying and improved shelf life
34
Amla, or Indian gooseberry (Emblica officinalis),
cultivated in India, has long been used in
Ayurveda.
Fruit contains between 700 and 1,000 mg of
vitamin C. The vitamin C is considered to be
highly stable due to the presence of tannins and
polyphenols.
Additionally, evidence suggests that amla may
help to boost immune response, presumably due
to its vitamin C content
Immunity: Emblica officinalis
35
Activating alpha waves in Brain: Theanine
Tea Contains Theaninie and Caffeine
Required Physiological Level – 50mg
Current Level – 20mg
http://www.biomedcentral.com/1471-2164/12/131
36
• Nutrilite is the vitamin brand to grow, harvest and process
plants on its own organic farms.
• Controls the development of its supplements from seed to
finished product.
• Claims to practice sustainable farming, the future of
organic farming.
• Owns 4 farms in 3 countries with over 6000 acres. (more
than any other supplement company)
• Established a Nutri-Certification program. This insures
that any plant concentrate purchased has been grown
according to strict organic farming specifications.
Primary crops: blueberry, Echinacea purpurea, Echinacea
angustifolia, oregano, peppermint, nettle, valerian, white
kidney bean, dandelion, lemon balm, black cohosh. spinach,
kale, broccoli, watercress, parsley, grapefruit, orange,
lemon, oregano, sage, and asparagus. acerola; picão preto,
also called Spanish needle, and passion fruit
37
Take Home Message
• To maintain normal human functions recommended calories
and RDA of micro + macro nutrients are required
• Nutrition can also enhance certain functions e.g. aging
• Rapid progress in biosciences pointing to altered genome functions by
nutrition
• Specially during pregnancy and early childhood
• A nutrition strategy therefore should consist of meeting RDA
and enhanced function
• Trees can play an important role : healthy carbohydrates and fats

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Nutrition Beyond Calories & the Role Trees can play

  • 1. 1 Nutrition Beyond Calories & the Role Trees can play Navin Sharma Science Seminar – 25 April 2014
  • 2. 2 • How knowledge is changing and how this will lead to future nutrition • Double burden of nutrition • Maternal nutrition and evolution of epigenetics • From single molecule to total calories – a case study of aging research • What will be the roles of trees? DNA controling you  environment controlling you  maternal nutrition  Mind controlling you?? Theme of the Seminar
  • 3. 3 Exogenous plant MIR168a specifically targets mammalian LDLRAP1: evidence of cross-kingdom regulation by microRNA. Zhang L et al. Cell Res. 2012 Jan;22(1):107-26 • Discovered plant micoRNAs in the serum of mice and humans. • The microRNAs possibly came from ingested rice. • The microRNA MI168a binds to the mRNA of low-density lipoprotein receptor adapter protein 1, inhibiting its translation. • Decrease LDL removal from plasma • Ingested plant microRNAs can regulate the expression of human genes. Micro RNA from food can be found in blood stream MicroRNAs (miRNAs) are a class of small noncoding RNAs that control gene expression by targeting mRNAs and triggering either translation repression or RNA degradation
  • 4. 4 Royal jelly is a complex, protein-rich substance secreted from glands on the heads of worker bees. To be queen larva is fed royal jelly inside a compartment called a queen cup. The workers and queens are genetically identical. The royal jelly diet makes the queen: develops ovaries and a larger abdomen for egg laying, The worker are sterile. The royal jelly silences a key gene (Dnmt3), which codes for an enzyme that silences a group of queen genes. When Dnmt3 is turned "on," the queen genes are epigenetically silenced, and the larvae develop into the default "worker" variety. To be Queen – need special diet Science. 2008 Mar 28;319(5871):1827-30. "Genetic Science Learning Center, University of Utah, http://learn.genetics.utah.edu."
  • 5. 5 Jerdon’s Jumping Ant Florida Carpenter Ant ANT GENOME SEQUENCING – WAY TO UNDERSTAND LONGEVITY • In 2010 – The Ant Genome sequenced • Jerdon‟s Jumping Ant & Florida Carpenter Ant • Very Different Social Structure • In Jumping Ant – Queen replaceable • New Queen Fed Special Diet – Lives 10- times Longer Up-regulation of telomerase and sirtuin deacetylases in longer-lived H. saltator reproductives, caste-specific expression of microRNAs and SMYD histone methyltransferases, and differential regulation of genes implicated in neuronal function and chemical communication Science (2010) – 329: 1068-1071
  • 6. 6 Malnutrition, per capita calories, Prevalence of diabetes and CVD Malnutrition Calories Diabetes CVD
  • 7. 7 Source: Global Health Risks (2009) Nutritional Double Burden
  • 8. 8 • Famine exposure in first 2 trimesters lead to 80% higher prevalence of overweight (p<0.0005) • Famine exposure in last trimester or famine exposure in first 5 months of life associated with 40% lower prevalence of overweight (p<0.005) Obesity in Young Men after Famine Exposure in Utero and early Infancy (Ravelli et al NEJM, 1976)
  • 9. The Fetal Origins or “Barker” Hypothesis • Pioneered by David Barker in the early 1990s – Painstaking development of historical birth cohort studies in England – Poor nutrition of mother during pregnancy: 2-fold increase in coronary heart disease from largest to smallest BW & Impaired GT increased from 14% to 40% • Impaired fetal growth causes adult disease • “Thrifty phenotype” - impaired fetal growth permanently changes the body‟s structure and physiology – Adaptive for compromised nutrition – Maladaptive for over-nutrition • Potential mechanisms include blood pressure & glucose tolerance Am J Clin Nutr 2000 71: 1344S-1352S.
  • 10. 10
  • 11. 11 Yellow Mouse Agouti Mouse Maternal Supplements with zinc methionine choline folate B12 •High risk cancer, diabetes, obesity •Reduced lifespan •Lower risk of cancer, diabetes, obesity • Prolonged life LTR Hypomethylated LTR Hypermethylated Hypermethylation leads to increase in lifespan and decrease in metabolic disorders Cooneyetal.(2002)JNutr.132:2393S Mother’s nutrition and epigenetic changes "Genetic Science Learning Center, University of Utah, http://learn.genetics.utah.edu."
  • 12. NUTRITION & HEALTH IN „OMICS ERA: EPIGENETICS • CpG- Cytosine Phosphate • Can be methylated to generate 5-methyl cytosine • Spontaneously deaminates to form Thymine • Poorly recognized by DNA repair mechanism • Thus CG  TG mutation propagates 12 Changes in gene expression, cellular memory, reversible, no change in DNA structure
  • 13. Homeostasis Broken Down with methylation Disease : Outcome of interaction of DNA and Food components
  • 14. Figure 4. Resveratrol induces overall hyperacetylation in HepG2 but not Hep3B or HuH7 cells. Venturelli S, Berger A, Böcker A, Busch C, et al. (2013) Resveratrol as a Pan-HDAC Inhibitor Alters the Acetylation Status of Jistone Proteins in Human-Derived Hepatoblastoma Cells. PLoS ONE 8(8): e73097. doi:10.1371/journal.pone.0073097 http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073097
  • 15. 15 Nutrient or diet Epigenetic mechanism Embryonic development Folate DNA methylation, imprinting Choline DNA methylation Protein restriction DNA methylation, histone modifications Alcohol DNA methylation Stem cell Butyrate Histone acetylation, DNA methylation Retinoic acid PRC Aging Folate DNA methylation Calorie restriction Histone acetylation Immune function Folate DNA methylation Cancer Methyl-deficient diet Histone modification, microRNA Genistein DNA methylation, microRNA (−)-Epigallocatechin-3-gallate DNA methylation, PRC Curcumin microRNA Obesity, insulin resistance High-fat diet DNA methylation, microRNA Methyl-deficient diet DNA methylation Curcumin Histone acetylation Inflammation Resveratrol Histone acetylation AdoMet Histone methylation Methyl-deficient diet microRNA Neurocognition Choline DNA methylation, histone methylation
  • 16. 16 Add life to age – clear evidence from foods and nutrition HEALTHY AGEING http://www.healthandage.com/html/min/eama/eama6/idx/docs/june_2006/TSAL_J_POWELL_Healthy_Ageing.pdf Oxidative stress appears to be a major determinant of life-span accessible to nutritional intervention.
  • 17. 17 SMR-Specificmetabolicrate Increase in SOD may act as a determinant in longevity ANTIOXIDANTS AND AGEING AmJC/inNutr1991;53:373S-9S MLSP- Maximum lifespan potential
  • 18. 18 ANTIOXIDANTS AND LIFESPAN Increased concentration of antioxidants help in longevity AmJClinNutr1991;53:373S-9S
  • 19. 19 Sod 2 null mutants show decreased life span in Drosophila Genetics165:2295–2299(December2003) A Sod2 NULL MUTATION IN DROSOPHILA
  • 20. Life-span and superoxide dismutase/catalase mimetics Small synthetic mimetics increases the mean life-span of C elegans by 44 %, and treatment of prematurely aging worms resulted in normalization of their life-span (a 67 % increase). MelovSScience2000Sep1;289(5484):1567-9
  • 21. 21 SOD 1 expression is higher in Male ant when compared to Queen SOD1 gene expression as measured by Northern blots Sod1 EXPRESSION IN ANTS SOD1 activity assay levels in activity units per milligram Lasius niger (Ant) Queens reach an age of 28 years, whereas workers for 1-2 years and males, a few weeks 3486–3489 PNAS March 9, 2004 vol. 101 no. 10 Oops!!
  • 22. 22 sod-2 mutant worms live significantly longer than wild-type worms PLoSGenetics:February2009|Volume5|Issue2 Sod-2 DELETION IN CAENORHABDITIS ELEGANS
  • 23. 23 EFFECT OF CR AND EXERCISE ON SURVIVAL IN RATS Adapted and modified from Holloszy 1995 ClinicalInterventionsinAging2007:2(4)537–543 Median lifespan is increased with Calorie restriction
  • 24. 24 Credit:UniversityofWisconsin-Madison/JeffMiller CALORIC RESTRICTION IN MONKEYS Caloric restriction slows ageing in monkeys
  • 25. Resveratrol (3,5,4‟-trihydroxystilbene) mimics the effects of dietary restriction • Hypothesized to activate Sir2, or SIRT1 in mammals • Alternatively, hypothesized to upregulate AMPK, downregulate mTOR (Mammalian target of rapamycin) Baur et al. 2006 Nature
  • 26. 26 ENHANCING ENERGY EXPENDITURE AND FAT METABOLISM Red grape (resveratrol) compound shows promise Industry looking for exercise mimetics now! www.medindia.net/news/tripepperblend
  • 27. 27 Basic Functional PrescriptiveCarbohydrates Fats Proteins Vitamins Minerals Water Flavonoids Omega 3& 6 Hunger Satiety Basic functions Driven by Cultural & Ethnic Preferences Preventive Performance Maintenance Specific needs For a condition Driven by Aspirational needs IndulgenceFunction Doctor Age Sustenance Enjoyment / Health Health & Wellbeing Well-being/ appearance Age Add life to years Evolution of Nutrition Deva Kumari, Yashwanth Radhakrishnan & Navin Sharma (2012): International J of Life Science & Pharma Res. 2(3):174-184
  • 28. Agriculture Life Sciences Forestry Nutrition Actives How to improve nutrition of people in Developing World 28 Epigenetics Human Genome • New ways of working • Newer / different alliances and partnerships • Local knowledge Focus in-and out-of-RDA
  • 29. Barley Max Odorless Soy RS in Wheat Amaranth Moringa Agroforestry – Complete Nutrition Nutrition Pyramid 29Methionine and Nyacin rich fruits: Peach, Jackfruit, Tamarind, Dates
  • 30. Synsepalum dulcificum, a West African berry better known as "miracle fruit," changes the way foods taste. Possible model for the taste-modifying activity of MCL. (A) MCL is held on hT1R2- hT1R3 at neutral pH (Left) and activates it under acidic pH (Right). Koizumi A et al. PNAS 2011;108:16819-16824 MCA= Miraculin MCL is widely used in Japan, where dedicated cafés are serving low-cal food with a miracle fruit on the side
  • 31. 31 Low GI Palm Sugar
  • 33. Oil with High Oleic / High Stearic Acid Margarines and spreads with cardio neutral saturated fatty acid – no chemical hydrogenation & no trans fats Enhanced functionality with higher heat stability for frying and improved shelf life
  • 34. 34 Amla, or Indian gooseberry (Emblica officinalis), cultivated in India, has long been used in Ayurveda. Fruit contains between 700 and 1,000 mg of vitamin C. The vitamin C is considered to be highly stable due to the presence of tannins and polyphenols. Additionally, evidence suggests that amla may help to boost immune response, presumably due to its vitamin C content Immunity: Emblica officinalis
  • 35. 35 Activating alpha waves in Brain: Theanine Tea Contains Theaninie and Caffeine Required Physiological Level – 50mg Current Level – 20mg http://www.biomedcentral.com/1471-2164/12/131
  • 36. 36 • Nutrilite is the vitamin brand to grow, harvest and process plants on its own organic farms. • Controls the development of its supplements from seed to finished product. • Claims to practice sustainable farming, the future of organic farming. • Owns 4 farms in 3 countries with over 6000 acres. (more than any other supplement company) • Established a Nutri-Certification program. This insures that any plant concentrate purchased has been grown according to strict organic farming specifications. Primary crops: blueberry, Echinacea purpurea, Echinacea angustifolia, oregano, peppermint, nettle, valerian, white kidney bean, dandelion, lemon balm, black cohosh. spinach, kale, broccoli, watercress, parsley, grapefruit, orange, lemon, oregano, sage, and asparagus. acerola; picão preto, also called Spanish needle, and passion fruit
  • 37. 37 Take Home Message • To maintain normal human functions recommended calories and RDA of micro + macro nutrients are required • Nutrition can also enhance certain functions e.g. aging • Rapid progress in biosciences pointing to altered genome functions by nutrition • Specially during pregnancy and early childhood • A nutrition strategy therefore should consist of meeting RDA and enhanced function • Trees can play an important role : healthy carbohydrates and fats