1. Leading nitric oxide research. Changing lives.
Nitric Oxide:
The Miracle Molecule
Nathan S. Bryan, Ph.D.
Chief Science Officer for Neogenis Labs
Professor of Molecular Medicine at the University of Texas Health Sciences Center
at the Houston Institute for Molecular Medicine
Disclosure:
N.S. Bryan is a paid consultant and
Chief Science Officer of Neogenis Labs, Inc.
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2. Leading nitric oxide research. Changing lives.
Nitric Oxide: Research
One of the Most Studied
Molecules In Medical History
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3. What is Nitric Oxide?
• A gas with chemical formula NO
• A free radical, reacting with the oxygen in air to form
nitrogen dioxide
• One of the few “signaling” molecules in the body of mammals
including humans
– Naturally produced in the endothelium
– Literally “tells” surrounding muscles to relax
– Enhances blood flow through proper dilation
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4. Thousands of Trials & Published Studies
Nitric Oxide
Molecule of the Year
Over 120K Articles
On Nitric Oxide
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6. Leading nitric oxide research. Changing lives.
Nitric Oxide: In the body
“The discovery of Nitric Oxide and its
function is one of the most important in
the history of cardiovascular medicine.”
The American Heart Association
Quoted in The New York Times
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7. Nitric
Oxide
Cell Proliferation
Apoptosis
Angiogenesis
Tumor cell growth
Immunology
Unspecified immunity
Inhibition of viral replication
Transplant rejection
Cardiovascular System
Vasorelaxation
Blood cell regulation
Myocardial contractility
Microvascular permeability
Peripheral Nervous System
NANC nerve-mediated relaxation
Central Nervous System
Learning/memory
Pain sensitization
Epilepsy
Neurodegeneration
Central BP control
Respiratory Tract
Bronchodilation
Asthma, ARDS
Gastrointestinal/
Urogenital Tract
Penile erection
Pre-term labor
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8. Nitroglycerine, the Original Nitric Oxide Donor
Smooth muscle
Endothelium
Plaque
Adventia
Atherosclerosis=plaques reduce blood flow
• decreases oxygen supply to the heart
• causes angina pectoris or myocardial infarction
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9. Nitroglycerine, the Original Nitric Oxide Donor
Smooth muscle
Endothelium
Plaque
Adventia
Nitroglycerine provides
NO to the narrow
atherosclerotic vessel
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10. Shear stress
Thrombocytes Monocytes Granulocytes
> anti-atherosclerotic properties
Regulation of vascular tone
and blood pressure
Inhibition of aggregation of
thrombocytes and of adhesion of
thrombocytes, monocytes and
granulocytes at the endothelium
Cardiovascular Effects of NO
Inhibition of migration and
proliferation of smooth muscle cells
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11. Leading nitric oxide research. Changing lives.
Nitric Oxide: Disease
Development
“NO plays an important role in the
protection against the onset and
progression of cardiovascular disease.”
2005, PubMed
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12. Endothelial Dysfunction is the Enemy
Insufficient Nitric Oxide levels =
endothelial dysfunction
Sufficient Nitric Oxide levels =
endothelial cells communicate with
surrounding tissues telling them to relax
Endothelial Dysfunction =
contributor to all major CV diseases
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13. RISK FACTORS
Hyperlipidemia
Arterial hypertension
Smoking
Age (45 males: 55 females)
Plasma NO concentrations reflect the degree
of endothelial dysfunction in humans.
Kleinbongard et al FRBM 2006
Plasma NO & Endothelial Dysfunction
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14. Nitric Oxide Production & Aging
Gerhard et al Hypertension 1996 Celermajer et al JACC 1994 Taddei et al Hypertension 2001 Egashira et al Circulation 1993
Adults over 40 make
far less Nitric Oxide
than their bodies need
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15. L-Arg
L-Arg
Arginase
ADMA
NOS
Uncoupling
Reduced Oxygen
Reduced Cofactor + Substrate
Oxidative Stress
Antioxidants
NO2
NO3
Oxidation
Bacterial Reduction
NO
O2
-٠۰
ONOO-
Disease
BH4
Ca/Cam
FAD+
FMN
NADPH
O2
Heme iron
GSH
Mitochondria
XO
NADPH
oxidase
Urea Cycle
L-Arg
Health
Diet
The Complex L-arginine Pathway
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16. How the road from promising scientific
breakthrough to real-world
remedy has become all but a dead end.
“From 1996 to 1999, the U.S. food and
Drug Administration approved
157 new drugs. In the comparable
period a decade later—that is,
from 2006 to 2009—the agency
approved 74. Not among them were
any cures, or even meaningfully
effective treatments, for Alzheimer’s
disease, lung or pancreatic cancer,
Parkinson’s disease, Huntington’s
disease, or a host of other afflictions
that destroy lives.”
Newsweek, May 15, 2010
The Current Environment
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17. Co-factor or Substrate Supplementation
L-Arginine
Ascorbic Acid
Folic Acid
Tetrahydrobiopterin (BH4)
Nitrosothiols
Nitrite/Nitrate
Nitro-fatty acids
Nitroglycerin/organic nitrates
NO hybrid drugs (NO-NSAIDS)
Require Functional NOS System
NOS-Independent Sources of NO
Ways to enhance NO availability
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Monday, June 20, 2011
18. Dietary nitrate is rapidly absorbed
into the bloodstream, where it mixes
with endogenous nitrate from the
NOS/NO pathway. A large portion
of nitrate is taken up by the salivary
glands, secreted with saliva and
reduced to nitrite by symbiotic bacteria
in the oral cavity. Salivary-derived nitrite
is further reduced to NO and other
biologically active nitrogen oxides in
the acidic stomach. Remaining nitrite
is rapidly absorbed and accumulates
in tissues, where it serves to regulate
cellular functions via reduction to NO
or possibly by direct reactions with
protein and lipids. NO and nitrite are
ultimately oxidized to nitrate, which
again enters the enterosalivary
circulation or is excreted in urine.
Human Nitrogen Cycle
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19. Bryan et al PNAS (2007)
Comparing mice on a low nitrite diet and
mice receiving supplemental nitrite
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20. Bryan et al PNAS (2007)
Heart attack injury is reversed
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21. *
*
*
Bryan et al, FRBM 2008
Steady state levels of NOx affected both
By diet and NOS
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22. Bryan et al, FRBM 2008
Nitrite Reverses MIR injury in eNOS-/- mice
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31. Nitrite is Enriched in Colostrum and Declines as Gut
Bacteria Colonize
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32. Traditional Chinese Medicine (TCM) has been used as a main stream of
medical care throughout Asia for centuries. But it is still considered an
alternative medical system in the western world. Mostly because of a lack of
understanding of their mechanisms of action and/or the active compounds.
Many cardiovascular diseases are characterized by a NO insufficiency.
There are a number of published reports on the association of TCM and NO-
related effects in cardiovascular field. However, their mechanism of action is
far from clear.
Traditional Chinese Medicine
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33. Tang et al FRBM 2009
Traditional Chinese Medicines for CVD
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34. Kinetics of NO Formation From Herbs
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37. Leading nitric oxide research. Changing lives.
Nitric Oxide:
Restoration
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38. Tibetans live at 4200 meter above sea level.
The low barometric pressure causes lower
arterial oxygen content and subjects the highlanders to
chronic hypoxia.
Study: Can Nitric Oxide be Naturally Restored?
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39. Erzurum et al.
PNAS (2007)
Results: Increased Blood Flow
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40. Because of increased NO production
Erzurum et al.
PNAS (2007)
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41. Leading nitric oxide research. Changing lives.
Nitric Oxide: Restoration
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Monday, June 20, 2011
42. Leading nitric oxide research. Changing lives.
Nitric Oxide: Restoration
Over 40
L-arginine cannot be
converted
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46. Significant reduction in patients
with elevated triglycerides
Neo40 Daily Clinical Trial Results
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47. Neo40 Daily Clinical Trial Results
Modest reduction in blood pressure
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48. Dosing & Protocol
Objective Dose Routine
Prevention, improved
absorption of dietary
nutrients & supplements
1 lozenge per day,
dissolve on tongue
Early in the morning on
empty stomach
Therapeutic, history of
CV disease or current
risk factors
1 lozenge, 2X per day,
dissolve on tongue
Early in the morning on
empty stomach, ½
before dinner on empty
stomach. At least 2
hours before bed.
For exercise
enhancement
1 lozenge 15-30 minutes
before exercise
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50. The First & Only Saliva Test for NO
Simple to use
Instant, easy-to-read results
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51. Testing Protocol
Baseline Neo40 Daily Test 1 Test 2
Repeat every
week
1 test on
empty
stomach
1 lozenge on
empty
stomach
15 minutes
later
2-3 hours
later
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52. Neo40 Daily Benefits
Benefit How it works
Energy Circulation
Sexual function Circulation
Sleep Circulation
Workout – pre &
post
Circulation
Mental acuity Circulation
Absorption of
nutrients
Circulation
Blood flow to
extremities
Circulation
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53. Chapter 3
NOtrition: NO-Boosting Foods and Supplements
Go For the Greens
“There’s a scientific revolution in progress,
and it’s happening on your plate.”
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54. Rich source of nitrate (beet root, artichokes, etc)
Contain the active nitrate and nitrite reductase, antioxidants
and polyphenols that can generate nitrite and NO
NO From Plants & Botanicals
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55. H. Garg, Master’s Thesis
Western Menu
Mediterranean Menu
Breakfast
Bagel w/ Cream Cheese Toast w/ Jam and Butter
Black Coffee (12 oz)
Cappuccino (Espresso+Milk)
AM Snack Carrot Nut Muffin
Yogurt
(Strawberry)
Diet Coke (12 oz)
Carrot Juice (12
oz)
Lunch
Big Mac
Mediterranean
Wrap
Large French Fries
Garden Vegetable Soup
Diet Coke (12 oz)
Mineral Water (12
oz)
PM Snack Snickers
Trail Mix
Black Coffee (12 oz)
Orange Juice (12
oz)
Dinner
Cheese Pizza (4 slices)
Salmon
(Smoked)
Diet Coke (12 oz)
Red Wine (12 oz)
Daily NO Intake Varies By Diet
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57. The Ultimate Nitric Oxide Index
Kale
6825
Swiss
Chard
2055
Arugula
1452
Spinach
1123
Chicory
938
Wild
Radish
814
Bok
Choy
775
Collard
Greens
697
Beets
632
Chinese
Cabbage
499
Le>uce
388
Cabbage
312
Mustard
greens
226
Cauliflower,
Raw
167
Parsley
150
Kohlrabi
136
Carrot
127
Broccoli
122
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58. Current paradigm for NO production through L-arginine is complex
and inefficient, especially in the aging population
Strategies to restore NO production/homeostasis will have a profound
impact on public health
Utilizing activity from natural products can overcome our inherent
inefficiencies and provide an essential source of bioactive NO
Rational design of NO rich herbs may provide a natural, cost effective
and over the counter product for conditions related to NO insufficiency
to improve health, performance and prevent disease.
Recognizing foods rich in NO potential may provide information into the
health benefits of certain foods/diets (Nitric Oxide Index)
Conclusions
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59. The University of Texas Health
Sciences Center at Houston
Harsha Garg
Yaoping Tang, M.D.
Ferid Murad, M.D., Ph.D.
Pamela Berens, M.D.
Albert Einstein College of Medicine
David Lefer, Ph.D.
John Elrod, Ph.D.
Mark Duranski
John Calvert, Ph.D.
Susheel Gundewar M.D.
LSU Health Sciences Center at
Shreveport
Karen Stokes, Ph.D.
Tammy Dugas, Ph.D.
D. Neil Granger, Ph.D.
Cleveland Clinic & Case Western
Cynthia Beall
S.C. Erzurum
Dennis Steuhr
J. Tejero
Acknowledgements
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62. Book Highlights:
Restoring nitric oxide
production in the
body thereby
combating:
• High blood pressure
• Heart attack
• Stroke
• Diabetes
• Arthritis
• Kidney disease
• Memory loss
• Osteoporosis
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