Physiological, Biochemical

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Physiological, Biochemical Molecular Aspects of
Aging

• Geoffrey Grant Ph.D.
• Professor of Science.
• University of Texas _at_Arlington

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My goal is to discover the factors that limit
life after reaching adulthood and to determine
biochemical, molecular genetic cause and effect
mechanisms that govern the aging influence on
lifespan.

• Develop therapeutic interventions.

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What is Aging?Correctly termed senescence
defined as the decay of the bodys homeostatic
maintenance after adulthood. Senescence is an
age/time-related deterioration of the processes
that control the functionality of the adult body.
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Longest Lived Human
Jeanne Calment _at_116 years Born Feb 21, 1875 Died
August 4, 1997 122 yrs old Arles, France
Senescence Progressive deterioration of the
bodily functions over time. Max. Lifespan
longest life greatest age reached by a member
of a species
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Topics of Discussion.
The Aging Process The Rate of Living
theory Oxygen Burden Electron Transport
System Free Radical Theory of Aging Reconcile
Theories Chemicals to rejuvenate cells and to
block and prevent oxidative damage
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Prediction of Genetically Pre-programmed
Life-Span
Predicted squaring of survival curves

• Between 1900 1950 acute diseases contributed to
  the flatness of the survival curves.
• Removing chronic diseases of old age will further
  extend the period of healthy life and create a
  SQUARED SURVIVAL CURVE.
• Robert N Anderson National Vital Statistics
  Report. Center for Disease Control 47 28 Dec.
  1999

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Survival of Calorie Restricted Rats

• Calorie Restriction (without nutritional
  deficiency) extends life in all species that have
  been tested.

• Eakin F Witten M. Experimental Gerontology
  1995, 30 33-64
• McCay CM et al. J. Nutrition 1935, 10 63-79

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Death Rate during the past

• Aging statistics start after 30 years old
• Re-Plotted Jones1955 In Handbook of Aging Chicago
  Univ. Press
• National Vital Statistics Report. CDC 1992

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Death Rates in the past century
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AGING MARKERS
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Performance Standard Vs AGE

• Performance standards set by world class athletes
• D H Moore Nature 253 264-5 1975
• P S Riegel American Scientist 69 285-290 1981

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Changes in Body Composition with AGE

• Lower lean body mass - less muscle
• Reduced protein synthesis
• Increased Abdominal Fat
• Lower fatty acid oxidation - available fat is
  stored
• Short KR Nair KS. J. Endocrin. Invest. 22
  95-105. 1999

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Hormone Levels Vs AGE
Growth Hormone

• Level determined by the 24 hr integrated GH
  concentrations in 80 men and 80 women .
• Zadik et al J. Clinn. Endocrin. Metab. 60
  513-516, 1985

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Life is a genetically programmed,
self-generating, self-assembling, bio-dynamic,
self-sustaining state. - production of energy
is both essential to life the inefficiencies in
energy production are detrimental and are a
primary cause of aging.
After birth organisms become totally independent
and totally self-reliant.
SO.
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THE Decline of Lifes Energy Theory of
AgingDOLE Theory.I am here today to frame an
argument that as cellular energy declines, bodily
functionality and performance suffers, until a
critical point is reached that can no longer
sustain the organism as a whole.This loss of
cellular energy, caused by oxidative damage, is a
primary cause of aging.
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Cellular Energy Production Vs AGE

• The consequence of the loss of mitochondrial
  function is progressive with age.
• NEEDS REF AND MORE DATA

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Energy Production Vs Cellular Processes

• Lower energy production levels results in lower
  protein synthetic rates.
• Loss of muscle and increase in fat storage.

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Life requires a constant supply of energy !

• The process of life is powered by energy as
  Adenosine Triphosphate ATP
• ATP can only be produced within each cell by
  intracellular mechanisms
• ATP drives the programming that produces
  development
• AND the maintenance of the adult.
• - production of energy is essential to life

-the inefficiencies oxygen utilization for energy
production are a primary cause of aging.
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Third Law of Thermodynamics.

• all systems in the absence of a sustaining force
  tend towards disorganization and,
  consequentially, entropy increases.

Life requires the receipt, conversion and storage
of energy this energy drives the genetic
programs. Epigenetic programs create
biological structures, organ inter-relationships,
and cellular and tissue organization during
your development.
AND ALL THE PROCESSES REQUIRE ENERGY INPUT.
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Third Law of Thermodynamics.

• LIVING systems are not, and cannot be, exceptions
  to the THIRD LAW.
• SO..

In the absence of a critical input of fuel and
the production of a critical level of energy
production as ATP the system fails. your
systems fail and your body fails
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Cellular Energy Production Vs AGE

• The consequence of the loss of mitochondrial
  function is progressive with age.
• NEEDS REF AND MORE DATA

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Mitochondria -site of cellular energy production
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Energy Production Loss
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Biological System Failure !
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METABOLIC RATE is an Energy measurement
Calories per gram per day
Rate of Living Theory The Higher the METABOLIC
RATE of an animal species the shorter the
lifespan. Metabolic rate can be measured as
Oxygen consumption. Decreases with increasing
body size or weight.
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METABOLIC RATE is an Energy measurement
Calories per gram per day
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Cutler Energy States !
Ln LSP X Ln SMR Ln LEP
LEP Humans 781 Other
Primates 458 Other Mammals 220
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Avian vs Mammalian Longevity
Comparisons from Metabolic Rate and Actual
Lifespan -Bats, Birds and Marsupials live longer
than anticipated
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Oxygen Demand for Energy Production !
http//www.sugar-bureau.co.uk/energy_zone/carbohyd
rates.html Carbohydrate Needs for Exercisers
1 hr/day 6 g/kg/d _at_ 60 kg body wt.
Glucose 360 grams/day 384 g Oxygen
http//btc.montana.edu/olympics/nutrition/eat12.ht
ml Fat Needs 1 g/kg/day _at_ 60 KG or 20 of
total calories as fat 60 grams/day Linoleic
Acid - C-18 168 g of Oxygen
Total oxygen demand for the utilization of 360 g
glucose 60 g Linoleic acid Oxygen requirement
550g /day
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The Bloods Oxygen Load !
Arterial Blood 96 saturated with
oxygen Venous Blood 64 saturated with
oxygen Each 100 ml release 6.5 cc oxygen gas
at Atm. Pres. Temp Oxygen Density 1.4
mg/cc 9.3 mg Resting Heart Pump Rate
100cc 60 beats per minute Resting oxygen
consumption 60 x 9.3 mg 559 mg
/min Oxygen per day 805 g
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Lifes Weakest Link !
Oxygen Load
From Glucose utilization 552g / day From
Blood Circulation 805g / day Total Oxygen
consumption 2- 300 kilograms/year
UNFORTUNATELY Oxygen Utilization is only 98
EFFICIENT
Reactive Oxygen Species ROS generation 2
4-6 kg of Hydrogen Peroxide/year Human
Lifespan 75.5 years average 75.5 x 5 kg
377.5 kilograms of H2O2 This forms THE FREE
RADICAL Theory of Aging
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Oxygen is required for life and intracellular
energy production BUT. Also is a root cause of
the aging process
Oxidative Damage by H2O2 and Reactive Oxygen
Species ROSwhich include superoxide radicals,
hydroxyl radicals,hypochlorous acid, peroxyl
radicals, and singlet oxygen.
O2 1e O2 - Ubisemiquinone reduced
Coenzyme Q10 donates an electron to Oxygen UQ10
- O2 O2 - UQ10 Non-enzymatic reaction
These highly reactive molecules are detrimental
to Cells and Mitochondrial structures and its
functionality.
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Oxygen is both required for life and is
detrimental
H2O2 is formed from the superoxide by an enzyme
called Superoxide Dismutase. 2H O2 -
O2 - H2O2 O2 AND broken down by Catalase
and Glutathione Peroxidase H2O2 --------gt
H2O H2O2 Fe2 OH - Fe3 The Hydroxyl
ion is a very highly reactive molecule reacting
in the order of 10 -9 seconds forming peroxide
with lipids, proteins, RNA and DNA
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Site of free Radical Production !
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Electron Transport System !
Electromotive Force drives the Formation of ATP
from ADP and Phosphate.
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Lifes Weakest Link !
Electro-motive Force drives ADP Phosphate to
Adenosine Triphosphate ATP
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nmol per min per mg protein
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Lifes Weakest Link !

• The adult bodys continuously utilizes Oxygen to
  produce energy within the cell.
• The Electron Transport Systems inefficiency
  produce free radicals Reactive Oxygen Species
  ROS that cause oxidation damage to cellular
  components.
• Lifespan depends upon-
• Metabolic rate - level of oxidative metabolism
• Efficiency of the Electron Transport System.
• The repair mechanisms in place.
• Sensitivity of tissue components to oxidation

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Protein Damage in Mitochondria
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Energy Production Loss
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Avian Efficiency !
Max. Lifespan Canaries 24 years
Parakeets 21 years Mice 3 1/2
years Pigeons 35 Years Rats 4
Years
Similar Metabolic rates and body size
G. Barja Ageing Research Reviews 1 (2002)
397411
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Mouse vs Parakeet vs Canary Heart Mitochondrial
Free Radical Leak
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Rat vs Pigeon Mitochondrial Free Radical
Production
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nmol per min per mg protein
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MOUSE VS BIRD Lipid Peroxidation tests
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Double Bond Index Diet Vs Tissue Lipids
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Bats Produce less Free Radicals
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Fatty Acid Double Bond Index vs Lifespan
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Rat vs Pigeon Mitochondrial Hydrogen Peroxide
Production
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Survival of Calorie Restricted Rats
Less ROS production in CR animals

• Calorie Restriction (without nutritional
  deficiency) extends life in all species that have
  been tested.

• Eakin F Witten M. Experimental Gerontology
  1995, 30 33-64
• McCay CM et al. J. Nutrition 1935, 10 63-79

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Longevous Compounds
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Longevous Compounds
ß-Alanyl-L-histidine
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Longevous Compounds
Primary Alkyl hydroxylamine
Secondary Alkyl derivative
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Effect of Primary Alkyl Hydroxylamine
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Effect of Primary Alkyl Hydroxylamineson Cell
Culture Population Doublings
Oxygen Concentration in Tissue Culture Incubator
Shay et al.
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Coenzyme Q vs Superoxide Production
nmol/min/mg Protein
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Coenzyme Q10 derivatives- Longevous Compounds
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Longevous Compounds
Cellular Oxidative Damage
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Mito Q and Cell Population Doublings
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Acetyl-L-Carnitine required for Energy Production
Acetyl Carnitine plus Lipoic Acid has been
shown to stimulate energy production and been
shown to extend the lives of treated
animals It improves in the quality of existence
by reestablishing energy metabolism.
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Oxygen Consumption and Ambulatory ActivityYoung
and Old Rats with without Acetyl-Carnitine-
30,days
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Mitochondrial Phospholipid Levels Vs AGE
and Acetyl Carnitine.
Cytochrome C Oxudase vs. AGE

• Cardiolipin formation in the membranes of the
  mitochondria requires the import of long chained
  fatty acids into the mitochondria, without
  cardiolipin fatty acids are not burned.
• This fatty acid transport requires the formation
  of a Fatty Acyl-Carnitine which needs carnitine
  to be readily available, may be supplemented as
  Acetyl-L-Carnitine.

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Phospholipids and Acetyl Carnitine treatment in
Rats
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Mortality Data and Acetyl Carnitine treatment in
Rats
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Energy Production Loss with Age
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Carnitine Treatment Restores ETC Enzymes
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Restores Electron Transport Chain
Enzymes. Enhances transport of fatty acids into
mitochondria. Increases level of Phospholipids
in membranes. -especially Cardiolipin Restor
es energy metabolism Improves Ambulatory
Activity of Animals. Improves Cellular
immunity. Changes ratio of Adipose tissue to
lean muscle mass in elderly.
Carnitine Acetyl-Carnitine Treatment
And reports of extension of lifespan in rats.
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Effect of Acetyl-L-Carnitine on Energy Production
ß-alanyl histidine dipeptide
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Survival of Calorie Restricted Rats

• Calorie Restriction (without nutritional
  deficiency) extends life in all species that have
  been tested.

• Eakin F Witten M. Experimental Gerontology
  1995, 30 33-64
• McCay CM et al. J. Nutrition 1935, 10 63-79

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Thank You
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8-oxo-deoxyguanosine levels vs Lifespan
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Relative DNA Repair
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INSULIN TREATED Calorie Restricted Rats

• Calorie Restricted reduction of Hydrogen Peroxide
  production is reversed by Insulin -2 week
  infusion.

• A.J. Lambert et al BBRC 316 2004
  1196-1201

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Survival of Calorie Restricted Rats

• Calorie Restriction (without nutritional
  deficiency) extends life in all species that have
  been tested.

• Eakin F Witten M. Experimental Gerontology
  1995, 30 33-64
• McCay CM et al. J. Nutrition 1935, 10 63-79

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Hormone Levels CR rats Vs Ad Lib Fed
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Respiration in Calorie Restricted and Ad Lib Fed
Rats
CR
Ad Lib
Ref DUFFY
Hours from onset of lights
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Survival of Calorie Restricted Rats

• Calorie Restriction (without nutritional
  deficiency) extends life in all species that have
  been tested.

• Eakin F Witten M. Experimental Gerontology
  1995, 30 33-64
• McCay CM et al. J. Nutrition 1935, 10 63-79