The Health/Life Extension vs.

Aging Disease Metabolic Pathway

as an Oppositional and Controllable Systematic Mechanism.
Gregory S. Bambeck PhD., Michael Wolfson J.D., M.B.A. and Warren Weller H. B.

Summary

The three most common and deadly diseases of aging are atherogenic
cardiovascular disease, the cancers and diabetes II, with obesity often being a
precondition for all three diseases. The caloric restriction (CR) metabolic pathway
conferring both health extension (HE) and life extension (LE) has just been recently
outlined. The three aging disease metabolic pathway systems run in direct
opposition to the CR/HE/LE pathway. The primary disease initiating defect(s) arise
from the fixation of an imbalance between anaerobic and aerobic carbohydrate
metabolism, most often precipitated by misdirected cell growth (CG) and/or CG
suppressor control elements, which in turn, are caused by either direct mutation or
external physiological regulatory control signal drive states gone awry. The overall
system is ancient, hailing back to even before the initial primordial eukaryotic cells.
We have generated a metabolic flow chart (map) that shows the CG/aging and
CR/HE/LE systems as an oppositional dynamic singularity. We make sense of these
diseases in the text, in terms of the metabolic map. Then, we show how disruptions
of proper metabolic map path flow are critical to the formation of these three
disease states. Throughout the discussion, we describe how the system can be
manipulated to alter their disease outcomes. Finally, from a compendium of food
supplements, we discovered a small handful of them that satisfied a quite stringent
activation site or map pathway mechanism of action set of criteria for avoiding and
fighting these diseases and accomplishing genuine HE, if not LE. The five most
successful candidates form a remarkable fit in terms of our metabolic map, while
the six runners up are very close fits or highly desirable system enablers. Used
singly, or in combination, all eleven have found efficacy in treating many dozens of
age related diseases beyond the three great killers of 85% of humanity we describe,
herein. Our arguments derive as much from carefully selected review papers as from
large scope work papers, so our references are broadly informative as well as deeply
technical, without overburdening the reader with a massive bibliography.

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Table of Contents

1. Introduction
2. The CG/CM/aging and CR/HE/LE System as a Metabolic Pathway Map
3. The Cancer Metabotype (CM) and Some Map Based Therapeutic Logics
4. Diabetes II, Metabolic Syndrome and the Map
5. Atherogenic Cardiovascular, Related Disease States and the Map
6. Food Supplements/Phytonutrient/Nutriceutical Candidates
7. An HE/LE Dietary Protocol for Even the Laziest of Minimalists
8. Conclusion
9. References

Contents topic 2 is mostly directed toward the research scientist, molecular

biologist or metabolic pathway specialist who wishes to keep up with current
research and its (and our) latest hypotheses. Contents topics 3, 4 and 5 are mostly
directed toward the above mentioned professionals and the medical professionals,
such as doctors, nurses, medical technicians and nutritional specialists. The
remaining contents topics (the introduction and from 6 through 9) are oriented
toward both the above mentioned professionals and the reasonably informed lay
person. All of the first six items in the contents represent our argument for personal
implementation of the minimalist dietary protocol, which should be of interest to
everybody. After all, our ultimate objective, is to arrive at a logical, straightforward
and simple protocol to induce genuine life extension.

1) Introduction

The most common and deadly diseases of aging are the 1) atherosclerotically
induced cardiovascular disease group, 2) middle age onset type II diabetes and 3)
the multitudinous array of cancers. Obesity might be considered a fourth disease, as
it shares many of the attributes of the metabolic pathway mechanisms of the first
three disease groups, but since its morbidity often precedes these three disease
states and manifests its ultimate outcome, in consequence, as one, two or all three of
these diseases of aging, it will only be treated, here, as a pre-condition. All these
diseases share, at their causative core, a carbohydrate utilization metabolic pattern

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of imbalances that are remarkably similar. Viewed superficially, this does not
appear to be the case, because these carbohydrate and related downstream
metabolic pathways operate in different physiological compartments and at
different levels of cellular, tissue, organ or organ system interaction. For instance, in
cancer, the metabolic pattern operates at the intracellular level as a direct and
stuck CG drive state we call the cancer metabotype (CM), and in fact, can trace its
origins to a single cell, often outwardly communicating only to its nearest
neighbors. Extracellular physiological manifestations of cancer mostly become
evidenced at the multicellular level, much later, as mesenchymal and metastatic 3D
tumor invasion and dispersal as its tissue mass causes increased signal strength to
its surrounding and, sometimes, distant tissues. Diabetes II requires the mobilization
of trillions of cells working in concert at the insulin generating and responding
tissue and organ level of integration; particularly between muscle, adipose, liver and
pancreas. Atherosclerotic cardiovascular disease also involves the participation of
gaggles of cells operating at the level of the intestine, liver, adipose tissue and
vascular tree. It manifests mostly as responses to inflammation induced by the
metabolic pattern itself and lipid carrier imbalances as altered liver, blood born and
arterial wall functions. The common denominator includes maladaptations to
carbohydrate metabolism that become fixed and self-exacerbating, ultimately
leading to each disease having characteristic forms of symptomatics and outcome.

Recent developments give us three great proofs that the aforementioned statements
are true. First, the same metabolic pathways and/or their downstream destructive
outputs are evidenced in all three disease situations. Second, and more importantly,
pharmaceutical, certain food supplement or phytonutrient metabolic pathway
differential rectifiers prevent, delay onset, delay progression, and in some cases,
actually reverse all three disease states with a single therapeutic regimen. It is only
very recently discovered (within the last three years) that these diseases, plus
critical aspects of obesity share such a surprising and striking metabolic
commonality at their very core. Lastly, pushing the metabolic system in the reverse
direction of its multiple disease manifestations with the same therapy actually
extends life beyond its normal maximum time span limit, which is defined as the
length of time beyond the time span enjoyed by the oldest 5% of the normal
population distribution, which can be an LE as high as 25-35% in mammals, and
translates into two to three decades in humans. This multi-serendipity is enough to

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induce a lustful hunt for a rational description for such an apparent Occams razor.

We have developed an essential skeletal outline of the entire regulatory metabolic

system with its most salient (and therefore, with minor omissions) inputs and
outputs. This map is provided in Figure 1, with legend. To simplify matters, it is
presented from the standpoint of a single cell. After describing its functions, we will
probe its CG/CM/aging vs. CR/HE/LE perturbations, which are most readily and
easily evidenced by cancer cells, because they represent the most straightforward
aberration of the CG system. Then, we will branch out to the somewhat more
obscure and less obvious whole body physiological manifestations evidenced by
alterations at the organ and tissue level as demonstrated by diabetes II and
atherosclerotic cardiovascular disease. Finally, we use these understandings, and the
map itself, to speculate on the mechanisms of action of both well and slightly less
understood pharmaceuticals and food supplement molecules that are known to have
had a beneficial impact on all three disease states, but have not had the benefit of
the cell growth and life extension pathways, to be matched to, until the present time.

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Figure 1. The cell growth (CG)/cancer metabotype (CM)/aging disease vs. the
caloric restriction (CR)/health extension(HE)/life extension (LE) pathway flow
system (map).

Circled ovals represent the core elements of the CR/HE/LE vs. CG/CM/aging
system. Boxes are used to contrast the mitochondrial CG neogenic state from its CR
driven regenic state, as shown here, in ovals. Neogenesis and regenesis are
described in the text. Asterisks represent very recently discovered or described
phenomena which made this map possible for the first time. All other (unboxed and
uncircled) terms represent well established knowledge gained prior to recent
asterisk ascribed knowledge. Most solid lines represent responses to stimulation,
while dashed lines show downstream impacts of a special core (RTG) catabolic
interaction. TSC2 and TOR (known as mTORc1 in mammals), are the trunk of the
input/output tree. The two opposing input branches flow downward through the
TSC2/TOR trunk and continue downward and outward from there, analogous to
roots. Input branches of the CR/HE/LE system are in the upper right hand
quadrant, while the functionally oppositional input branches of the CG/CM/aging
system are in the upper left hand quadrant. The same quadrant concept is not true
for the bottom half of the diagram because input dependent opposing outputs of
TOR have differential impacts upon all of its targets. Arrows represent up
regulation and blocking end plates represent down regulation. All components are
best thought of as if in the activated state, when observed alone and upstream
effects are ignored. The map should be kept at the ready when reading the text,
because the two are inextricably co-dependent. In the text, we show how CR, HE
and LE are the same pathway, while CG, CM and aging are also their own self-same
pathway, but in opposition to CR, HE and LE. A legend is provided, below.

Legend: (in alphabetical order)

AKT- the protein kinase AKT

AMP-adenosine monophosphate
AMPK- adenosine monophosphate kinase
Angiogenesis- process of initiating vascular growth toward a tissue
Apoptosis- programmed cell suicide mechanism
ATP- adenosine triphosphate

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COX-2- cyclo oxygenase 2
4E-BP- eukaryotic translation initiation factor 4E binding protein
EGF- epidermal growth factor
Genotoxic stress- generally refers to DNA damage necessitating ROS reduction
system and DNA repair mechanism initiation
GH- growth hormone
HIF- hypoxia inducible factor
I and IR- insulin and insulin resistance
Mitochondrial neogenesis- making new, but ox/phos incomplete mitochondria
(described in text)
Mitochondrial regenesis- the process of completing ox/phos in neogenic
mitochondria (described in text)
IGF- insulin like growth factor
Inflammatory cytokines- molecule which initiate the inflammatory reaction,
either intracellularly, extracellularly or both
NF-kB- nuclear factor of kappa chain activated B cells
P53- tumor suppressor p-53 protein
PGC-1alpha- peroxysome proliferator activated receptor gamma (ppar gamma)
co-activator-1alpha
P13K- phosphoinositide 3-kinase
PKC- protein kinase C
RAS- rat sarcoma virus protein
ROS- reactive oxygen species
RTG- mitochondrial/glycolytic retrograde response
SESN- Sestrin (a redox containing active domain protein)
S6K- ribosomal protein S6 kinase
TGF- transforming growth factor
TOR- target of rapamycin (mTORc1 in mammals)
TSC2- tuberous sclerosis complex 2
VEGF- vascular endothelial growth factor

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2) The CG/CM/aging and CR/HE/LE System as a Metabolic Pathway Map

This segment of our narrative is presented in a rather concentrated and rapid fire
fashion. A solid understanding of metabolic pathways and a reasonable familiarity
with their regulatory elements is very helpful. For those less inclined, we
recommend that you skip forward to the three disease type examples in contents
topics 3, 4 and 5 and/or to sections 6 through 9. For those who wish to brush up a
bit before jumping in, we recommend that you read the metabolism Special Section
introduced by L. Brian Ray, Science, vol. 330, p. 1337, 12/3/2010; Z. Feng, Cold
Springs Harbor Laboratory Press, p.199, 2010; Seyfried and Shelton, Nutrition and
Metabolism, vol. 7, no.7, 1/27/2010 [1, 2, 3] This is up to date review material that
is highly focused around the subject of this article, but without the unifying
metabolic pathway system. It is helpful if one has a printed copy of the Figure 1
CG/CM/aging and CR/HR/LE metabolic map on the side, as a reference guide,
when reading this paper, as we refer to the map throughout the narrative. We will
begin with an overview of the connection between the aging diseases from a
generic, understanding of the map before delving into details.

In the mid twentieth century, it was discovered that calorically restricted (CR) mice
lived longer than their normal ad libitum maximum life expectancy, as previously
defined toward the end of the introduction, and which we term as true life
extension, just life extension or LE, for short. CR is usually defined as holding an
organism to 40% below ad libitum caloric intake, while maintaining nutrient
balance. CR is initiated by elevated AMP concentrations resulting from exhaustion
of importable glucose as a consequence of caloric restriction. AMP directly
activates AMP kinase kinase (AMPKK) to activate AMPK, then TSC2 to inhibit
TOR, which shuts down anabolic genes while activating genes to increase catabolic
efficiency. Also, the cell responds to glucose need and paucity by increasing its
sensitivity to insulin (I), and activating regulatory proteins that inhibit the CG
system. An activated CG system has the opposite outcomes as CR. Since then,
research has reported the CR/LE phenomenon throughout all major branches of the
proto-animal and animal kingdom, from fungi, worms, insects, spiders, fish, and in
mammals, from rodents to primates. The field lay fallow for decades, until just
recently, when LE was also noted to occur when using the anti-tissue rejection
pharmaceutical rapamycin and the anti-diabetic drug metformin. Metformin

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activates AMPK to inhibit TOR, while rapamycin directly inhibits TOR; two
actions shared with CR. Knowing their pharmacological targets and actions on
those targets provided the earliest inklings of a pharmaceutically induced up
regulation of the bulk of the same LE pathway genes activated by CR. For the first
time, the thousands of genes activated by CR had been whittled down to a few
target regulatory proteins that seemed to induce a similar set of gene activations and
protein products. We call these drugs and similarly acting food supplement
molecules, CR mimetics. All CR mimetics cause a statistically significant delay in
the onset of aging diseases, a circumstance we call health extension, or HE.
Unfortunately, it is too early yet, to tell exactly which HE food supplements actually
might induce LE because not enough time has elapsed to complete the experiments,
nor has there been enough grant money to support the vast array of experiments
needed. However, a few of the many thousands of possible herbal and food
supplement candidates share metabolic pathway mechanisms of action that
similarly or directly mimic the regulatory pathway systematics of CR and the LE
pharmaceuticals, and all known mimetics operate on the TOR inhibition input side
of our map, whether as direct CR component activators or as CG component
inhibitors.

Even more recently, on the opposing CG side of the TOR input tree, cancer
investigators found powerful tumor CG inhibiting properties in the molecules that
strike at the highly activated catabolic core glycolytic pathway downstream of TOR
and common to all known CG upstream TOR pathway activating factors. Previous
therapies had focused on the CG factors, themselves. Similarly, CR pathway
activation increased insulin sensitivity, reduced hyperglycemia, slowed CG,
inhibited tumor growth and inhibited the whole downstream gene transcription
output of the normal CG pathway by inhibiting TOR, thus, contradicting the cancer
CG pathway, allowing us to connect the CR/LE and CG systems into a mutually
oppositional regulatory interaction in terms of some of the critical metabolic
features of diabetes II, and allowing us to revisit an old cancer cell metabolism
hypothesis.

A new and dawning realization in the cancer research community, that mutations in
the CG proteins controlling core metabolism caused cells to remain stuck in a CG
metabolic pattern and inhibited the CR metabolic pattern, resurrected a long

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forgotten more than seventy year old hypothesis put forth by Otto Warburg.
Warburg stated that cancer cells differed from normal cells, in that they are stuck,
both during and between rounds of replication, in a state of aerobic glycolysis in
which anaerobic glycolysis is dramatically enhanced and aerobic mitochondrial
respiration (as reflected in oxygen consumption) is dramatically reduced, thus
forcing carbohydrate metabolites to be funneled into lactic acid export and anabolic
cell growth rather than efficient catabolic energy generation. Normal cells, on the
other hand, are not stuck and could shift back and forth so as to efficiently couple
glycolysis to respiration by maintaining highly capable respiratory function between
rounds of replication. Unfortunately, the respiratory deficiency portion of the
hypothesis was proven false (or more accurately, not always true) some 56 years
ago. Also unfortunately, all of Warburgs good postulates were thrown out with the
one that was false. Twenty years later, in 1976, the error was corrected by G.
Bambeck in his mitochondrial alteration dissertation at Kent State University, when
he attempted to direct attention from respiration deficiency toward the entirety of
the mitochondrial oxidative phosphorylation (OX/PHOS) system, which includes
the electron transport chain, the chemiosmotic proton pump via mitochondrial
intramembraneous space sequestered protons from NADH and FADH and their
coupling to ATP synthetase, in addition to oxygen consumption, so as to show how
such alterations were integral to supporting both reducing power and metabolic
substrates for a metabolite flux change that assisted an anabolic drive state [30].
However, the Warburg rejection was so firmly entrenched, that he was ignored at
that time, and incidentally, for the next 30 years.

It is only during the last three years that the Bambeck modification has been shown
to be true and to be part of a natural consequence of profound cell cycle disruptions
in respiratory/OX/PHOS and metabolite flow. Although the author is still ignored,
the idea is definitely not. This was mostly realized when the relationship between
CG drivers through TOR was found, soon after the turn of the twenty first century,
to directly affect the relationship between glycolytic and mitochondrial function.
Our new understandings of modern genomic, proteomic and metabolomic
molecular biology rigorously reinforce these ideas, and Otto Warburg is back on
center stage, albeit very posthumously. Even the December 2010 issue of Science
(referenced earlier) admitted that the wholesale rejection of Warburg had been a
multi-decade blunder of massive proportions, and dedicated a multi-paper mea

9
culpa, in somewhat of an apology. Although science sometimes makes big mistakes,
the intrinsic nature of its methodology eventually corrects them. In this paper, we
refer to this metabolically CG stuck state of the cancer cell as the cancer
metabotype (CM).

Thus, we were enabled to generally connect CG, CR, LE, cancer and diabetes II into
a more global unification. Atherogenesis was less obvious, but became an easier fit,
when understood from the CG/CM-CR/LE context, as a result of direct pollutant
ROS (such as smoking) activation of the NF-kB to inflammatory cytokine system,
and/or CR pathway inhibition and CG pathway activation, primarily focused on the
liver (particularly fatty liver) and arterial wall, in relation, mostly, as a response to
obesity in a way not too dissimilar from diabetes II. Obesity causes a large increase
in circulating I and IGF, causing activation of the cell survivor factor system all
over the body, which, in turn, feeds directly into all of the bodys organ and tissue
CG systems. This physiologically reinforces activation of all three aging diseases,
and most horribly, doubles the incidence of the most common cancers (G. Taubes,
Science, vol. 335, p. 28, 1/6/2012) [4]. Results published in the March 2010 issue of
Science [10] helped integrate much of this overview into a singularity by knitting an
antioxidant sestrin redox containing active domain protein (SESN) to the tumor
suppressor protein P53 and into the CR/LE pathway, to be more deeply discussed
and referenced, later. We will also describe, later, how it was necessary to divide
mitochondrial biogenesis into two functionally discrete domains in order to render
the pathway unification hypothesis consistent, coherent and realistic. All of the
above combined to yield the structural outline for the metabolic flow diagram
shown in Figure 1.

What is not shown in the chart is that there is considerable reinforcing regulatory
self-talk and counter reinforcing cross-talk between the two pathway systems. This
means that activating regulatory stimulators of one pathway synergistically co-
stimulate multiple components in the self-same pathway, while inhibiting control
elements in the opposing pathway. This permits us to keep the map both reasonably
accurate and simple, while avoiding confusing clutter, and hopefully, over
simplification. The chart also does not include wholesale nuclear gene system
activation and deactivation, as they are described in the text, as needed. Thus, the
basic perspective of the presentation is from the cytoplasm of a single cell to the

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extracellular milieu via the plasma membrane. After all, it is the cytoplasm which
contains the vast bulk of intermediary bioenergetic metabolism. Also, it is important
to note that most of our presentation will be based on mammalian physiology. Now,
we delve deeper into the system.

The system provided in the flow chart is an outline of the regulatory control
pathways monitoring, responding to and governing catabolism, anabolism, nutrient
availability, cell energy status and cell damage and repair integrity; all being in
terms of the cells decision to grow and divide under CG drivers, or to hunker
down and wait out something as severe as a famine, or even something as small as
an overnight fast. First, we will look at the system from the cell growth side of the
equation. Cell growth and division is critical for life, as organisms need to manifest
it, explosively, during fetal development and wound healing, while utilizing a more
leisurely rate for worn out cell replacement. We represent it, here, mostly from the
perspective of a fetal cell, because it so closely resembles cancer, but in a regulable
form. For instance, cell growth and replication, in the fetus, advances at a furious
pace, is in a hypoxic environment, needs to invade surrounding tissues, requires
vascularization and parasitizes the fuel and nutrient base that is provided carte
blanche by the mother (or host), similar to an aggressive cancer.

Growth factors, like growth hormone (GH), transforming growth factor (TG),
epidermal growth factor (EGF) and, to date, more than twenty known others,
stimulate either the rat sarcoma virus oncogene protein (RAS) and/or P13K, via
their activated kinase and tyrosine phosphatase cascades to up regulate the protein
kinase AKT (AKT) to block TSC2, to TOR activate the hypoxia inducible factor
HIF, ribosomal S6 protein kinase (S6K) and the mitochondrial neogenesis portion
of mitochondrial biogenesis via PGC-1alpha. S6K activates the thousand, or so,
genes that up regulate the whole of the anabolic system. PGC-1alpha activates the
thousand, or so, genes for the biogenesis of OX/PHOS incomplete neogenic
mitochondria and participates with its co-activator ppar gamma to shut down lipid
catabolism derived acetylCoA for entry into the Krebs cycle. This favors lipid
synthesis and the ATP production requirement becomes much more favored by
heightened anaerobic glucose metabolism via transcription of the low Km fetal
glycolytic enzyme set ordered by HIF. The terminal enzyme in glycolysis, pyruvate
kinase, becomes replaced by a fetal form that causes a build up of glycolytic

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intermediates, particularly the three carbon molecules, glyceraldehyde 3 phosphate
(G3P), dihydroxy acetone phosphate (DHAP) and the six carbon glucose 6
phosphate at the headwaters of glycolysis, all of which drives into the pentose
phosphate shunt, (PPP) for formation of the pentoses in nucleotide synthesis to
create the information molecules, such as DNA and RNA, and the bioenergetic
carriers such as ATP. G3P and its interconvertible analogue DHAP, also form the
three carbon backbones for triglyceride synthesis and also, their penultimate
breakdown. The lipid synthesis motif also can be co-opted by CG activators to form
fatty liver and adipose tissue, in addition to enhancing cancer cell replication, as we
shall see later. For now, it is sufficient to note that glycolysis, PPP, gluconeogenesis,
lipid synthesis and lipid catabolism are joined at the G3P hip as one might say, a
commonality which implies an ancient heritage. It might be noted here, that G3P is
also a multi-faceted core intermediate in the photosynthetic Calvin cycle of plants,
and that this pathway shares many substrates and reaction intermediates with
gluconeogenesis and PPP. In all of these most primitive of central metabolic
pathways, proton and electron shuttling are employed to create and lose energy,
while chemiosmotic proton electromotive force is not employed, or to put it in a
more evolutionary context, was not yet likely, employed, back in the day.

One can actually see the evolutionary roots of animal life in the three carbon sugar-
phosphate analogues (particularly G3P) and of higher sugars as energy carriers in
the glycolytic, PPP and their immediate accessory pathways, and particularly, their
integration with the nucleotide energy carrying and information molecules with
their head to head dinucleotide pyrrophosphate redox water-hydrogen and carbon
dioxide-oxygen substrate couplers NAD, NADP and FAD, on an ancient anaerobic
earth, when cellular bioenergetics was ruled by substrate phosphorylation, prior to a
life sustaining concentration of photosynthetically derived oceanic and atmospheric
oxygen. In essence, the modern animal cell, just like its ancient counterpart,
becomes a highly anaerobic ATP producing sugar fuel junkie, and appears to
revert back to a more ancient catabolic format as part of its replication process, so
as to preserve amino acids, fats and nucleotides as new cell construction material. It
is as if some metabolic form of ontogeny recapitulating phylogeny hails back to
the days when the anaerobic host cell was evolving replicative control over its
acetate feedstock requiring aerobic invader, which purportedly came from the alpha
proteobacter evolutionary lineage, as seen through genomic analysis, and then is

12
thought to have finally become the mitochondrion. It is probably from prior to this
evolutionary heritage (in anaearobic prokaryotes) that the regulatory
phosphorylation and kinase cascades that dominate CG were derived, then later
followed by the aerobic prokaryotic acetylation and deacetylation metabolic protein
regulation required for the later integration of sugar fed anaerobic glycolysis and
acetylCoA fed oxidative mitochondria to orchestrate efficient metabolic substrate
flow, redox substrate needs, bioenergetic partitioning and co-ordination of the
eukaryotic cell cycle. We will revisit the acetylation/deacetylation issue via
metabolomic and proteomic analysis, later. It is gratifying to evolutionary theory
that these systems and their components still remain so clearly visible today. We
now leave our little evolutionary side bar, to return back to HIF function.

In addition to its profound effects upon glycolysis, HIF also blocks the apoptotic
mechanism, which would normally be poised for activation by the elevated
mitochondrial ROS output under these conditions. ROS would also activate SESN
to activate p53, but p53 is inhibited by CG regulatory elements. HIF also activates
vascular endothelial growth factor (VEGF) that is exported from the cell to
mitogenically stimulate vascular endothelial cells to grow capillaries, followed by
arterioles etc. to feed the growing and dividing CG activated cells with oxygen, fuel
and nutrients, in a process called angiogenesis. Angiogenesis is important in wound
healing, fetal growth and cancer tumor growth. Paradoxically, under the apoptosis
resistant CG drive state, most probably as a result of elevated genotoxic stress, the
mitochondrion becomes primed with pre-apoptotic proteins that sensitize the
mitochondrion to increasing its outer membrane permeability (MOMP) for
cytochrome c release, which in turn, institute the apoptotic cascade. This is more
pronounced under the CM state than the CG state, because the order to die or
switchover to a CR regimen is either not given or not received, as it is in normal
cells. MOMP in the CM state is demonstrated, and with interesting therapeutic
results by Triona Ni Chonghaile et.al. Science, vol. 334, p. 1129, 11/25/2011 [5].
This issue will also be revisited when we look at cancer therapeutics. A sterling
review integrating autophagy, the mitochondrion, apoptosis and MOMP functions is
provided by D. R. Green et. al., Science, vol. 333, p. 1109, 8/26/2011 [6].

To support the growth system for impending tissue invasion, P13K and TOR
stimulated S6K activates the release of cytoplasmic bound NF-kB to the nucleus,

13
where inflammatory cytokines such as cyclo oxygenase 2 (COX 2), tumor necrosis
factor (TNF) and interleukins (ILs) become transcribed and exported to initiate
extracellular edema, interstitial matrix breakdown and a furthering of the
inflammatory cascade. In the interests of speed, the system sacrifices cleanliness
and efficiency by elevating ROS generation, inhibiting self-component recycling
via autophagy, and by delaying the 4E-BP activated transcription gene set for
nuclear mitochondrial respiratory/OX/PHOS proteins until after the CG driver
system has been shut down. These late gene set respiratory/OX/PHOS proteins
render inefficient neogenic mitochondria into efficient regenic mitochondria. The
autophagy we just mentioned, although integrally tied to metabolism, is mostly
activated/deactivated by regulatory elements of the CG and CR systems, and is too
complex for discussion, here. An excellent review of autophagy and the metabolic
regulatory control elements is provided by J. B. Rabinowitz and E. White in
Science, vol. 330, p. 1344, 3/12/2010 [7]. Suffice it to say, that the autophagy
regulatory network is a sterling example of CG and CR regulatory element self
pathway reinforcement and cross pathway inhibition. Also under CG activation,
hexokinase II associates with the external surface of mitochondria to highjack
mitochondrial ATP to entrap phosphorylated glucose within the cell immediately
following importation, and to reinforce hyperactivation of the glycolytic/PPP
system. Mitochondria can also adapt to anaerobiasis by importing deaminated
glutamine for substrate phosphorylation via the succinyl CoA synthetase step,
therefore, bypassing citrate synthetase, which normally utilizes the glycolytic post
end product, acetyl CoA to prime the Krebs cycle. Glutamine importation excess is
not terribly uncommon in cancer cells, but seems most pronounced in the minority
of cancer cell types operating at the lower end of an elevated glycolytic rate,
seemingly, to reinforce proper CM driven substrate flow (A. J. Levine and A. M.
Puzio-Kuter, Science, vol. 330, p. 1340, 3/12/2010) [8]. Thus, we can see how the
core anaerobic and aerobic catabolic systems complement the anabolic drive state
initiated by the CG/CM pathway.

Mitochondrial participation in CG has always been a bone of contention,

particularly in the case of cancer cells, and even more so since the 1956 demise of
the Warburg hypotheses. The recent elucidation of the CR/LE pathway interaction
with the CG/CM pathway, by ourselves and others, has helped to resolve this
debate. A very clean and recent paper by Z. Szijgyarto et. al., Science, vol. 334, p.

14
802, 11/11/2011 [9] measures all of the most salient variables in terms of the
relevant bioenergetic impacts of the anaerobic/aerobic catabolic matrix via the
inositol phosphates IP6 and IP7, as CG downregulators via AKT inhibition. They
demonstrate this elegantly in both yeast and mouse embryonic stem cells. They
measured all ATP/AMP, NAD/NADH, glycolytic rate and mitochondrial
respiration/OX/PHOS rates and coupling, in both the IP6-IP7 up and down
regulated states, and found all of our aforementioned glycolytic and mitochondrial
CG and CR effects in type quantity and sequence. The real significance of their
work is that they conducted a time line focused multi-functional and targeted, but
limited, metabolomic analysis of an up and down CG drive state that is also
requisite and manifest, respectively, as an oppositional down and up CR response
state from the standpoint of relative glycolytic and mitochondrial bioenergetic
output responses. It has been known for a long time that IP6 is an anti-aging disease
and HE adjuvant, but its conversion to IP7 as a more powerful AKT inhibitor had
been unknown. Also, the pathway and its impacts had never been so fully and so
temporally articulated. One real powerful take home lesson is that, from the
standpoint of CG and CR, the mitochondrion has very bipolar behavior. This
contrast is further reinforced when we compare the oppositional activation and
functions of PGC-1alpha and 4E-BP.

Mitochondrial biogenesis has long been thought of as a singular process, initiated

principally, via PGC-1alpha, even though it has been known, for over two decades,
that mitochondrial biogenesis consists of two transcription phases, aptly named, the
early phase and the late phase. The early phase begins immediately after TOR
activation of PGC1-alpha via phosphorylation, and produces the bulk of the
thousand, or so, proteins that make up the vast majority of the proteins constituting
the mitochondrion. Indeed, from all outside appearances, a whole new
mitochondrion is born; but, in fact, such a mitochondrion is very functionally
incomplete. The late phase proteins are a small, but vital set of deactivated TOR
block lifted eukaryotic transcription initiation factor 4E binding protein (4E-BP)
mitochondrial respiratory chain proteins also required for OX/PHOS, typically
produced some 24 to 48 hours after mitosis, and have been traditionally thought to
merely complete the mitochondrial biogenesis process. However, until this process
is completed, the mitochondria are not fully coupled, nor do they become fully
coupled until CG has ended and lifted the TOR block on 4E-BP. This mitochondrial

15
second phase lag is seen throughout fetal development, and has traditionally been
thought of as a mere consequence of rapid proliferation and, therefore, as a
component in a singular biogenic process. We see it as a dual process with the first
phase requiring the absence of the second phase if cell growth and proliferation is
to be able to occur at all. More importantly, we see the cancer cell, unlike its normal
counterpart, to be interminably trapped in the CG/CM neogenic state and
chronically suffering from a regenic deficit. We are not saying that the cancer cell
has no mitochondrial regenic capacity, because, as we shall see later, there is reason
to believe that CG and CR drive states may have temporary and limited
oppositional subroutines that leave cancer cells in a somewhat steady state of
mitochondrial OX/PHOS dysfunction.

Thus, the elucidation of the CR/HE/LE system, TOR as a toggle switch oppositely
activating the early and late phase aspects of mitochondrial biogenesis and the
different roles of mitochondrial catabolism during CG opulence and CR starvation,
as the reader can see, forced us to split mitochondrial biogenesis into distinct
functional phases we call mitochondrial neogenesis and mitochondrial regenesis.
Neogenesis is driven by CG, makes new OX/PHOS inefficient mitochondria and
participates in CG metabolically. When CG shuts down and/or CR turns on,
regenesis creates late gene mitochondrial OX/PHOS efficiency, and switches to
functioning in the between replication cell quiescence, cleaning and maintenance
phase. The CG pathway shows this, and its oppositional CR pathway, described
below, further illustrates the point.

The CR/LE pathway upstream of TOR is best described by starting at AMPK. Until
a mere year ago, AMPK was primarily viewed as a cell energy status sensor, being
up regulated by elevated concentrations of energy poor AMP, and being down
regulated by its absence, as a result of high concentrations of its alternative energy
rich form as ATP. AMPK is now also known to participate as a powerful reactor to
ROS and genotoxic stress from ROS effects via P53 to SESN, or other SESN
activation, as shown in our chart. From the CG vs. CR standpoint, the largest ROS
generator is via poorly coupled mitochondria derived from the CG drive state, as
previously described. Together, these components act as a combination of
mitochondrial neogenic or regenic function feedback sensor, as well as a nutrient
availability monitor, both a CG or CR drive state switch activator, and an energy

16
status an ROS production sensing and response cluster that acts through TOR. The
SESN/P53/AMPK/TOR/4E-BP vs. PGC-1alpha interactions are brilliantly
articulated in a work paper by Jun Hee Lee et. al. in Science, vol. 327, p. 1223,
3/5/2010 [10].

When activated by cell energy depletion via a low ATP/AMP, or an elevated ROS
activated SESN or P53, AMPK activates TSC2 and, thereby, inhibits TOR, which
lifts its block of 4E-BP and initiates transcription of the 4E-BP late phase
mitochondrial regenesis respiratory/OX/PHOS complex genes. These regenerated
mitochondria become efficient in OX/PHOS and dramatically cut ROS production,
which in turn, reduces genotoxic stress, and shuts down p53 induced DNA repair
mechanisms. This also requires that there is no strong CG driver, but work with CR
mimetics shows that it can and does overpower cell survivor factors, such as I and
IGF. Such therapy has remarkable impacts upon the diseases of aging, particularly if
they are obesity associated, as obesity is a powerful I and IGF promoter, as
referenced previously. A CR to AMPK activation state eventually lifts TOR
inhibited autophagy (referenced earlier) causing the cell to engage in frugally
scavenging its own defective parts for the new famine-like situation maintenance,
energy and repair requirements. We must note here, that if uncoupled mitochondrial
ROS damage becomes too severe for scavenging and repair, an already cross
pathway CR sensitized apoptotic mechanism due to activated p53, cytochrome c
efflux from dying mitochondria, a down regulated CG pathway and others, will
institute cell suicide. Since HIF is CR down regulated under these circumstances,
the low Km glycolytic fetal enzyme system reverts to the slow and efficient
glycolytic rate that, in turn, slows down PPPs anabolic contribution, lactic acid
production, and with other CR system regulator outputs, such as angiogenesis,
anabolism, neogenesis, the inflammatory response etc.

The map also shows that mitochondria and glycolysis talk to each other through
metabolic intermediates and not yet fully understood factors, mediated in part by
NF-kB and COX2, in a complex system called the retrograde response (RTG). This
helps facilitate the switching back and forth between mitochondrial metabolic states
associated with regenesis and neogenesis in the context of up and down regulation
of glycolysis. In general, the RTG response is utilized in the on position by
hypoxic neogenic mitochondria with poor oxygen utilization when cells are highly

17
dependent on anaerobic glycolytic substrate phosphorylation for ATP, as is found in
fetuses and in the core of neoplastic tumors. Interestingly, in cancers, mitochondrial
telomerase can relocate to the nucleus to initiate immortalization. It is intriguing
that central elements of both cell suicide (apoptosis-cytochrome c) and immortality
(mitotic index-telomerase) were both evolved to be released from mitochondria.
Thats a lot of power for such a lowly organelle. A much deeper treatment of RTG
and mitochondrial function is provided by Seyfried and Shelton, as referred to
previously.

It would be so easy if everything just feeds into and out of TOR as a simple toggle
switch. However, nothing in biology is simple. There are many TOR molecules in a
cell, and they can be geographically and microsomally compartmentalized in the
cell, with some in the on position in one compartment, while others are in the off
position, elsewhere, as was initially revealed in the spring of 2011 and further
refined in November 2011. It appears that both TOR conditions might be operating
in a pseudo homeostatic condition, when there is no clear cut drive state, such as
during circadian rhythms, when the system leans CG during post prandial states or
leans CR during temporary, light fasting, sleep states. The circadian CG and CR
states are clearly and succinctly described by J. Bass and J. S. Takahashi, Science,
vol. 330, p. 1349, 3/12/2010 [11]. In clear cut drive states, like starvation or host
provided ad libitum fetal growth, the TOR system is much more likely to respond
wholesale to the preponderance of the inputs. This paradox of two possible TOR
states appears to have been resolved by R. Zonca et.al., Science, vol.334, p.678,
11/4/2011 [12].

They found an additional TOR activation, via a membrane attached lysosomal

internal amino acid abundance detection mechanism. This activating strategy may
have provided us with the hint of a CG subroutine that can operate within the CR
program, when autophagy and mitophagy nutrients become available, as this TOR
activation format acts as a lysosomal internal amino acid detector and mobilizer. In
short, a lysosomally external membrane bound ATPase-RAG complex recruits
inactive TOR from the cytoplasm to become Rheb activated when the lysosomally
enriched amino acid stimulated ATPase proton pump acidifies the lysosomal lumen
and dissociates the ATPase-RAG complex to activate TOR. This system is
surprisingly independent of the CG via the AKT/TSC2/TOR pathway, as it can

18
drive lysosomal membrane bound TOR activation in cell and cytoplasm free
lysosomal suspensions simply as a result of lysosomal amino acid loading and
ATPase activation alone. Thus, TOR can be activated outside the conventional CG
drive state. However, nutrient availability must be present, even for this alternative
activation.

Lysosomal amino acid loading can occur via extracellular importation or from
autophagy. Autophagy is inhibited during CG, while fuel importation is externally
constrained during CR. Conversely, autophagy is up regulated during CR,
particularly when readily available internal food stores run low and internal
component recycling becomes necessary. Autophagy comes to the rescue by raiding
defective internal resources and by providing nutrient energy rich and metabolic
building block autophagosomes that merge with lysosomal elements and bud off
into lysosomes. It appears that all of the correct components have appeared in time,
space and circumstance to elicit a cellular housecleaning and efficiency upgrade
with scavenged resources, just ahead of a worsening and possibly, critical famine:
everything the cell needs, for sustenance and repair, except for an impossibly
unavailable external food supply, that is.

A similar to TOR, but even more obfuscate situation has emerged with the
conundrums associated with the silent information repeat gene product (SIRT 1
lysine deacetylase). To those who have been watching the so called life extension
scene for the last several years, SIRT 1 lysine deacetylase (hereafter called SIRT 1)
was once hailed as the greatest hope as a CR activated LE control element. For one
thing, it seemed to be activated by the same LE mimetic molecules that activate CR,
and it, itself, is activated by CR. More so, SIRT 1 responds to the depressed
NADH/NAD that occurs when reducible substrates become in low supply, as
happens under CR conditions. However, experiments with organisms containing
extra copies of SIRT 1 genes or hyperactivated SIRT 1 genes yielded mixed results,
ranging from shortened life to just HE. A brief history of the unraveling of the SIRT
1 story is provided by J. C. Frankel, in Science, vol. 334, p. 1194, 12/2/11 [13].
Other revelations, concerning the actual SIRT 1 molecular functions in the CG vs.
CR pathways, demonstrate why SIRT 1 is so glaringly (and probably genuinely
surprising to many), absent from our metabolic map.

19
The timing and function of SIRT1 activation, relative to the timing of CR, AMPK,
AKT and PGC-1alpha and their functions, in the whole, are excellently articulated
in a small but well presented set of work and review papers by E. H. Jennings et.
al., Oncogene, vol. 29, p.1056, 8/19/2010; C. Canto et. al., Nature, vol. 458, p.
1056, 4/23/2009 and N. B. Ruderman et. al., Am. Jour. Physiol., vol. 298 no. 4, E-
751, April 2010 [14, 15, 16]. Altogether, the results show SIRT 1 to actually be a
CG activator!

From a timing perspective, the response of AMPK to the lack of bioenergetic ATP
availability via increased AMP/ATP is a fast response that can happen in seconds, to
minutes, to hours depending upon the speed type and severity of CR. One may
envision cyanide poisoning, an insulin induced hypoglycemic episode or post-
prandial sleep as examples, respectively. Regardless, the AMPK response can be
fast, and is fast when AMP rises. SIRT 1 responds to a much more slowly
developing situation in the cell. Much later, after AMP levels rise in the cell, the
redox potential begins to collapse as the NAD/NADH rises to a critical level. It is
important to note that if AMPK is activated at X time units after elevation of
AMP/ATP, then the NAD/NADH activation of SIRT 1 may take as long as one to
two orders of magnitude longer than X time units. The next question is: What is
SIRT 1 doing when it does it?

For one thing, an over expressed SIRT 1 activates AKT by deacetylating it and
causes a CG mode AKT activation that up regulates TOR, increasing tumor
formation and shortening life span (Sundarsen et. al., Sci. Sig. ra 46, July 2011)
[17]. SIRT 1 has also been shown to be a 12 site deacetylase and activator of PGC-
1alpha, while AMPK is a phosphorylator and inhibitor of PGC-1alpha. PGC-1alpha
is known to have activation/inactivation dual contrapositive functions. In its CG
activated state, it institutes mitochondrial biogenesis (neogenesis) and shuts down
its ppar gamma co-activator fatty acid catabolism association, while in the AMPK
to TOR CR drive state, it shuts down building mitochondria, associates with its co-
activator ppar gamma and initiates the catabolic fat conversion to mitochondrially
required acetyl CoA that is needed due to the loss of pyruvate from sugar deprived
glycolysis. The acetylation and deacetylation of the epsilon amine of proteinaceous
lysines actuating the dual contrapositivity of PGC-1alpha, is referred to as its yin-
yang function. Just as TOR may have a CG-like function during CR, SIRT 1 may

20
even have a more pronounced, but similar CG function. Perceiving TOR and SIRT
1 as having a normal ability to function as CG subroutines during CR removes the
paradoxes and conundrum of function surrounding them. It also explains how cells
can slowly accumulate new mitochondria under CR conditions, when PGC-1alpha
is not under its pronounced activation commanded by the CG drive state. Further, it
explains how mutationally or genetically altered over expression of SIRT 1 can
shorten life and increase cancer incidence.

From all these data, we have formulated the following CR temporal sequence
hypothesis: As CR institutes elevated AMP/ATP, AMPK is activated. Cross pathway
regulation inhibits the P13K, AKT, TSC 2 to TOR CG axis. Inhibited TOR shuts
down all anabolic S6K, HIF/glycolytic and mitochondrial neogenesis from PGC-
1alpha via its TOR kinase deactivation while AMPK phosphorylates and down
regulates PGC-1alpha in preparation for SIRT 1 deacetylase function, which if
needed, may come later. TOR deactivation turns on the 4E-BP mitochondrial
regenesis program to diminish ROS output and increase ATP production efficiency.
Simultaneously, PGC-1alpha switches off mitochondrial neogenesis and turns on its
lipid recruitment and lypolysis program to provide acetyl CoA feedstocks into the
Krebs cycle to make up for falling glycolytic input and output, and thus, the
eventual exhaustion of readily available internal food stores. As these stores exhaust
and autophagy becomes induced, NAD/NADH rises and SIRT 1 deacetylation is
activated as a CG subroutine of CR. SIRT 1 engages CG activation of AKT and
PGC-1alpha via deacetylation to assist autophagy provided lysosomal food store
activated TOR, to assist in a punctuated CG subroutine for anabolic
macromolecular assembly, and mostly bypass anabolic metabolite production via
the glycolytic and PPP anabolic and reduction support pathways, which at the very
least, suffer from both external and internal feedstock limitations. The SIRT 1
deacetylated PGC-1alpha from one of the halves of yin-yang function, temporarily
reactivates the mitochondrial neogenesis program to build a small contingent of
neogenic mitochondria from recycled cell parts derived from lysosomal TOR
activation by substrates from a now matured autophagy and SIRT 1derived AKT
activation via deacetylation. In addition, SIRT 1 temporarily stunts AMPK and p53
activity and erects its own cytoplasmic ROS fighting program (possibly through
FOXOFOXO gene activation) in p53s stead. Soon after re-establishment of
NAD/NADH redox potential, and the now prepared to become additional capacity

21
for conversion into regenic mitochondrially derived energy efficiency, the SIRT 1
subroutine shuts down and the p53/SESN/AMPK function set re-establishes
standard CR system master control. PGC-1alpha is reacetylated through the
contrapositive other half of the yin-yang process via GCN transacetylases and AKT
is also re-acetylated, so TOR shuts down and 4E-BP is reactivated, and thus,
mitochondrial regenesis ensues, assuring efficient low ROS ATP production with
maximum energy output and minimal fuel expenditure. One might wonder if the
systems critical energy needs might be supported by residual 4E-BP mitochondrial
regenesis proteins still lying around, at the ready, from the immediately previous
CR. In our hypothetical scenario, the SIRT 1/TOR CG subroutine cycles back and
forth like a metronome, as needed, within the CR master program. In a sense, the
CR machinery turns like a wheel, or better yet, like a spiral, going from a CR
dynamic to a special form of CG subroutine, and then, back around to a second and
weaker CR dynamic, again, with all of this occurring under CR conditions. Round
after round of CR-CG-CR continues to attempt to retain homeostasis for as long as
possible with minimal external fuel until the cell either limps along interminably,
exhausts itself to death in defect ridden apoptotic starvation or locates exogenous
food and breaks the cycle. Thus, we come to the end of our hypothesis. This
hypothesis plus our first evolutionary side bar leads us to our second evolutionary
side bar.

The epsilon amine acetylation/deacetylation of selected protein lysines has gained a

lot of scrutiny recently, because of the discovery of its over 2,000 protein
modification set in eukaryotic cells, with many of the proteins being regulatory in
nature. Historically, acetylation was thought to be restricted almost entirely to the
function of selective histone anionic loosening on DNA for large orchestrated gene
set transcription in eukaryotes, with deacetylation having the exact opposite effect.
Protein acetylation/deacetylation was also considered to be virtually non-existent in
prokaryotes, which also turned out not to be true. Because of the large protein sets
involved, most of the data has been collected by proteomic partitioning analysis
followed by amino acid composition back tracking to the genome, so as to
determine evolutionary relatedness and cellular function compartmentalization.

We have selected a few excellent representative papers that, together, yield an

overview of some of the early conclusions that seem to be emerging, such as papers

22
by: J. Patel et. al., Nutrition and Metabolism, vol. 8, no. 12, 3/3/2011; Kun- Liang
Guang and Yu Xiong, Trends in Biochemical Sciences, 2010; Hirschey Lab,
lab.hirschey.org/research/files/acetylation, 2012; J. Zang et. al., Molecular and
Cellular Proteomics, vol. 8, p. 215, 2/1/2009 [18, 19 ,20, 21]. In the prokaryote
model, as typically represented by E. coli, there are about 100 lysine acetylated
proteins with over 50% devoted to metabolism, over 20% devoted to translation and
the remaining minority devoted to various known and unknown functions. In
prokaryotes, the genetic conservation appears to hail back to a putative alpha
proteobacter thought to be the forerunner of the mitochondrion. Notably, the vast
majority of the metabolic contingent is utilized during the switchover from and to
fermentable and non-fermentable substrate catabolism. Also, in prokaryotes, the
acetylation/deacetylation system can undergo these sweeping changes with the
activation of just a single phosphatase and phosphokinase.

In eukaryotes, the largest plurality of the acetylome is also metabolic, and is very
DNA sequence conserved from the prokaryote acetylome, while the remainder of
the eukaryotic acetylome has radiated outward to include virtually every facet of
cellular function. It generally appears that the primitive host archaeobacter and its
pre-mitochondrial prokaryotic passenger had much mutual integration to attend to
in orchestrating their two genomes, proteomes, metabolomes and cell cycles in the
process of evolving into true eukaryotes, and it appears that much of this arose from
the use of the (same as prokaryote) non-fermentable mitochondrial feedstock, acetic
acid.

The net result from this hybridization, that led to Kingdom Animalia, represents a
quantum leap in complexity and variability, and that it led to even us humans, who
can sit around and ponder it all. In general, these preliminary results, with the
inclusion of what was said in our first evolutionary side bar, indicate that
acetylation is a temporal evolutionarily handmaiden to phosphorylation, from the
protein regulatory side of the equation. We present this notion less so as an
hypothesis and more so as conjecture. We await further acetylomic and genomic
analysis of even more ancient and strictly anaerobic prokaryotes to see where the
very origins of protein phosphorylation and acetylation might lie and/or have
diverged.

23
In conclusion, the metabolic map and the text look at the CG and CR metabolic
control systems in the large and from a somewhat holistic perspective. However,
we will always need meticulous experiments that decipher the specifics of
individual protein-protein, protein-nucleotide, protein-substrate, and other,
functional interactions. These component specifics are the ultimately deduced
mechanistic particularities that make up systems. But then again, to arrive at a
deeper understanding of the overall operations and meanings of the system as a
whole, we will need more input/output and temporal experimental design analyses
that arise from the combination of the genomic, proteomic and metabolomic (the so
called, shotgun analyses), that see everything at once. Only by looking at all the
interactions simultaneously, like a helicopter crew looking down over the traffic
patterns of a large city, will we gain understanding and control over its complex
integrated entirety. We have been very fortunate over the last decade, that these
tools have become mature, rapid, incredibly cheap on a per data point basis, and
that the databases connecting them together have become more fluid and integrated.
Surely, exciting times are ahead. We now turn our attention to focus on the medical
implications of the metabolic map and the possibilities for understanding of, and
intervening in, several of humanitys major killing diseases.

3) The Cancer Metabotype (CM) and Some Map Based Therapeutic Logics

As we saw from the previous section, the glycolytic and mitochondrial metabolic
pathways form a catabolic core system driven by CG and CR. Furthermore, these
systems make coherent and consistent requisite changes depending on the cell state
in all animal organisms and all of their cell types, with the major core cell system
initiating variations operating at the CG and survival factor level on the plasma
membrane. No matter what the status of myriad regulatory elements superimposed
on the ancient core system, it must respond in a flexibly limited fixed manner, or
the global system collapses. Apoptotic and novel metabolic cancer therapies show
some of the bounds of this limited flexibility. In normal cells, the core system can
toggle back and forth between the CG and CR states. It cannot do this, either easily,
or at all, in the cancer cell, as a result of mutations in the core system or in the
regulatory elements of the core system. We distinguish this mutationally stuck
cancerous growth state from the normal CG state by referring to it as the cancer
metabotype, or CM, for short. It has long been felt that the core system was a mere

24
normal handmaiden to cell growth factors and other regulatory elements, and
therefore, unworthy for investigation as a means of chemotherapeutic attack. Recent
developments show that many types of cancer cells can be growth arrested, killed or
even differentiated by direct attack on the core system, its most intimate regulators
or nearest neighbor/function partners. However, cancer cell kill therapies are neither
the thrust, nor the scope of this paper, albeit the logic behind such therapies is well
within its purview. Our primary focus is to define the basic CG/CM and LE/CR
systems, show their relatedness to principal aging diseases and to conduct a search
for phytonutrient regulators which rectify the system in a way that helps to prevent
or renormalize the major diseases of aging, and which promote HE and/or LE.
Thus, in this segment, we will give a cursory overview of cancer in terms of our
metabolic flow chart, and some of the therapeutic logic that can impact cancer.

The cancers are a madhouse of mutagenic realities in terms of both their many cell
variations and types, and the myriad ways they can convert CG to CM. For
instance, there are over twenty growth factors, and even more growth factor
receptors and downstream cascade components that are specific to different tissue
types and cancers, each of which would need a different magic bullet if attacked at
this level. As an example, Herceptin blocks the up regulated growth promoter
estrogen receptor on breast cancer cells and is, therefore, a magic bullet for this
kind of flaw. This receptor is not the error for most breast cancers, or virtually any
other types of cancer, for which, Herceptin is useless. The amount of research time,
cost and testing to develop such a vast target specific array, is mind boggling. Later
mutations in the cascades down mutatstream of growth factors and their receptors
can, and do, eventually circumvent such therapy. There are just too many branches
and cascade components funneling into the CG system, to thwart. Even insulin
dependent cancers can skirt therapies by up regulation of non-insulin dependent
glucose transport proteins, such as GLUT 4, to feed CM hyperglycolysis. CR cant
even significantly slow down the growth of these tumors [4]. We must note that the
high ROS index of cancer cells fosters further mutagenesis to enhance selective
survival of individual cells from the tumor cell population. Many tumors contain
several, or more, clonal subpopulations in a tumor mass. However, all of these CG
errors facilitate the core metabolic CM system. Even as we move deeper into the
CG vs.CR and closer to a smaller cadre of TOR inputs, therapeutic prospects look
grim, while the preventative aspects are sterling, as will be discussed later.

25
On the CR/LE side, the P53 tumor suppressor protein is defective, in one of many
possible ways, in about half of all cancers, being far and away, the single most
common gene product functional inducer of cancer, but we have yet to figure out
how to fix defective proteins; and even if we did, we are still in the infancy of our
understanding of how to target gene and gene product therapy to the cell type of
interest. In addition, too much of a good thing might be a bad thing, as mice with
extra copies of P53 genes die prematurely. In addition, p21 and PTEN, two tumor
suppressor proteins activated downstream of p53, are also often mutated, with
PTEN actually being absent from nearly half of all tumors. In total, this really
restricts the cells ROS fighting, CR pathway activating and CG suppressing
capabilities. Back on the CG/CM side, RAS function is also elevated in a plurality
of tumors and provides a reasonable therapeutic target, but can be circumvented by
P13K, which is the most often activated protein on the CG side of the equation, and
can become activated by survival factors. To make things even more complex,
PTEN actually inhibits P13K, so its absence even exacerbates the CG situation.
Also, about half of all tumors are CG driven by elevated I, IGF and/or enhancement
of their receptors, particularly if circulating I and IGF have been elevated by
obesity. This is the primary CG drive state accentuating mechanism by which
obesity doubles the rate of incidence of cancer, and increases the risk of diabetes II
by as much as an order of magnitude, particularly if the diet is overloaded with
sugar [29]]. One, or more, of these upstream of TOR CG protein or tumor
suppressor protein defects is found in virtually 100% of all cancers, implying that a
stuck CG drive state is, at least, one of the clearest hallmarks of cancer.

Attacks at this level are more like cancer management than cure strategies, as they
basically renormalize metabolic flow into a non-growth dynamic, which might
sometimes initiate their stem cell progenitor format to differentiate into a less
mitotic form, or delay disease progression rate by slowing growth, which is not a
bad thing when one has a lethal disease, but it is neither a prevention of future
mutational circumvention nor a cure. The HIF to VEGF response family, which is
downstream of TOR, could possibly be a much more useful target as it is strongly
impactful to glycolysis, angiogenesis and apoptosis. Blocking HIF would shut down
the whole fetal enzyme driven glycolytic pathway while the rest of the TOR
activated anabolic drive state would still be operating, cross pathway inhibitory

26
response notwithstanding. Not only would this block PPP, but it would block the
cytoplasmic ROS quenching effects of its products NADPH and glutathione. This
would really throw a monkey wrench into the whole CM metabolite flow dynamic.
As a bonus, this also blocks angiogenesis downstream of VEGF, which shuts down
the tumor fuel supply, in addition to down regulated HIF already having diminished
its ability to use a fuel supply, by down regulating glycolysis. To sweeten the pot
even further, blocking HIF also sensitizes the cell to apoptosis, and since the CG
drive state would still be operating through TOR, neogenic mitochondrial ROS
production should further sensitize apoptosis. The closer our therapies get to the
CM glycolytic core system of metabolism, the more general the impact across a
broader array of tumor types should be, particularly if glycolysis can be selectively
down regulated while the rest of the cell is stuck in a CG drive state.

Some recent developments in glycolytic attack logics have really raised a lot of
eyebrows. Blocking Hexokinase II with 3-bromopyruvate eradicated a hepatoma in
all cases of a rat tumor model. Attacking HIF induced fetal pyruvate kinase 2
(PMK2) with dichloroacetic acid has shown merit against a broad array of tumors .
Dichloroacetate disrupts CM gylcolysis by renormalizing pyruvate flow into
mitochondria, supposedly by restoring PMK1 or PMK1-like activity. This removes
the CG through HIF typical cancer cell PMK2 blocking of pyruvate production
which, in turn, causes glycolytic intermediates to pile up and activate PPP (A.
Coghlan, New Scientist, p. 6, 5/15/2010) [22]. Some cancers merely stop growing,
while others just shrink, or even, go away altogether. PMK2 is not just a glycolytic
enzyme that converts phosphoenolpyruvate to pyruvic acid, as it also acts as a ROS
sensor that inhibits its pyruvate formation function when oxidized, causing elevated
glycolysis to initiate the PPP production of cytoplasmic ROS reduction products,
such as NADPH and glutathione during CG when neogenic mitochondria are at
their maximum ROS generating state. Conversely, PMK2 is upregulated by the anti-
oxidant, glutathione, but its maximum activity is still only half that of PMK1.
PMK2 also associates with HIF and moves to the nucleus to assist in the HIF
activation gene set (D. Anastasiou, Science, vol. 334, p. 1278, 2/12/2010) [23].
More work needs to be done to determine how these variable tumor outcomes,
although all favorable, are achieved. It would seem wise to broaden our assault
further up the glycolytic chain and its input branches to the PPP. There are many are
simple molecules to mimic, and there should be thousands of inexpensive

27
candidates. Even as long as thirty five years ago, 5-thio-D glucose, which is a
potent killer of cancerous cells but not normal cells, achieved its desired in vitro
therapeutic goals by glycolytic disruption. Maybe we should look into that and
similar areas of work, again, and with our more modern tools. The fact that the vast
array of pre-TOR inputs funnels down to the somewhat inherently linear pathway of
glycolysis that is attackable in a medically favorable way, lends strong support for
the aerobic glycolysis portion of the Warburg hypothesis. It is good news that we
have robotic micro-array tissue culture and automated fluorescent cell status and
cell apoptosis monitors that allow testing thousands of molecules and dozens of cell
types at a rapid pace.

Therapeutic attacks on mitochondrial functions can also be envisioned. In fact, very

recently, two have been tried, and been found to have a respectable degree of
efficacy. T. N. Chonghaile et. al. [5] looked into a MOMP therapy and its effect on
the excised bone marrow cancer cells from myeloma and other cancers. They
figured that since cancer cells are unusually close in proximity of tumor cell
mitochondria to the apoptotic threshold, a process called mitochondrial priming,
but dont die because apoptosis is blocked by HIF and associated pathways, that
tipping those cells over the priming edge would kill them. Then they developed an
in vitro assay to monitor mitochondrial priming. They tested their hypothesis with
anti-cancer chemotherapies that induce apoptosis and found that strongly primed
mitochondrial cells responded well while poorly primed cells did less so. They also
found that normal cells were spared, having little measurable priming. Although this
result is not directly related to any metabolic function, the fact that mitochondrial
ROS induced priming and apoptotic inhibition are strongly linked to CG via HIF
and aerobic glycolysis, supports the efficacy of our metabolic map and its
functional description.

An even more recent publication by M. Jain et. al., Science, vol. 336, p. 1040,
5/25/2012 [24], shows that they have discovered a way, to at least, dramatically
reduce the intermediate term growth rate of the fastest growing cancer cells. Their
metabolomic study followed the consumption and release of 219 metabolites across
60 cell lines from the National Cancer Institute NCI-60 panel. They discovered that
the fastest growing cell lines had a very abnormally elevated activity of the
mitochondrion specific serinehydroxymethylase transfer enzyme (SHMT2) that

28
converts serine to glycine, then shuttles it to a direct PPP accessory pathway for
purine synthesis. This assists an already elevated glycolysis to PPP accessory
pathway for enhanced pyrimidine synthesis. Together, this increases the rate of
nucleotide synthesis. The most rapidly growing cancer cells, when inhibited by an
anti-SHMT2 inhibitory RNA , (RNAi) were immediately caused to nearly stop their
growth by almost 100%, and that about 85-90% of this inhibition was retained for
three days, which translates to about three to five doubling times, depending on the
cell line. This is a clear cut case of a mitochondrial control element defect further
entrenching aerobic glycolysis. We got a bit of a chuckle out of one of their
conclusions. They kind of scoffed at hypothesis driven research, preferring their
own massive number of components data driven research, then they conclude with
the hypothesis that SHMT2 might be a reasonable in vivo therapeutic target for
further research. Regardless, metabolomics is a very useful and powerful tool set. It
might be noted, that back in the late 1970s, there was also some promising in vitro
work with the OX/PHOS disruptors rhodamine 123 and tetraphenylphosphonium,
but we dont know if it was followed up on. There was little to no funding for
Warburg related research in those days. OX/PHOS disruptors and uncouplers are
also somewhat toxic to normal cells, but they enhance apoptosis. Perhaps they
might be useful as adjuvants with apoptotic sensitizers.

Interesting synergies between glycolysis and mitochondrial therapies can be

envisioned. TOR up regulation of both systems keeps mitochondria spending most
of their existence in a neogenic default state. In some cases, this is concomitant with
a large glutamine importation increase, but it always includes a programmed
OX/PHOS insufficiency. Glutamine deamination inhibitors or Kreb cycle blockers
downstream of oxalosuccinis acid, could inhibit glutamine pathway flow and assist
glycolytic disruptors in further catabolic substrate flow disregulation with TOR
activated anabolism [8]. Used judiciously, respiratory chain blockers or OX/PHOS
to ATP uncouplers could also disrupt CM flow integrity.

Inducing CR and its generic cross pathway moderate inhibition of CG regulators

can have a significant growth inhibiting effect on glucose plus I or IGF CM driven
tumors, which is partly due to the intracellular metabolic impact on glycolytic
substrate flow and partly due to it causing a decrease in circulating systemic I, IGF
and glucose [5]. The impact is far less in I independent tumors. Either way, the

29
outlook is very grim. However, CR and CR mimetics cause an incredible two-thirds
reduction in the incidence of cancer in the first place, and across the entire panoply
of solid tumors. Actual reduction in the disease state, as well as a similar to cancer
whopping reduction in incidence, is much more commonly found in atherogenic
cardiovascular disease and diabetes II, when using CR or CR mimetics. Following
these pursuits would cause an incalculable reduction in human suffering and the
national medical bill.

The number of papers attesting to the multiple aging disease impacts with CR
mimetics, across the years, is enormous, even though the researchers had no idea
that they were studying CR mimetics, because 1) they had never heard of the
concept of CR mimetics, 2) they did not know of the mimetics molecular targets, nor
of the targets CR impact and 3) they had no concept of the existence of a CR
pathway and its relationship to CG. All they knew was the outcomes of their
experiments and the net up and down regulation of the peripheral pathways (of CR
and CG) they measured. Actually, knitting it all together is very straightforward,
now that we know all of the above. Such research goes back decades and the papers
number in the so many hundreds, and maybe even thousands, that it is a waste of
time to reference them. It turns out to be a veritable treasure trove of CR/HE/LE
data, and it is relatively easy to mine. All one needs to do is search engine our
selected food supplement candidate molecules from section 6 and the disease type
from sections 4, 5 or 6 of this paper to become overwhelmed (Internet search
engined under: curcumin, silibinin, berberine, resveratrol, pterostilbene, epigallo
catechin gallate (EGCG), genistein, inositol 6 hexaphosphate, omega 3 oil,
coenzyme Q or alpha lipoic acid and cancer, diabetes II, cardiovascular disease or
obesity: 11 chemicals x 4 disease/conditions = 44 combinations in all) [25]. The
Life Extension Foundation probably has more such references than any other single
web site in the world (lef.org/references) [26].

From all that was presented thus far, we conclude that Warburg was correct, in that
aerobic glycolysis is a hallmark of cancer, but he was incorrect in stating that a
mitochondrial respiratory defect and the resultant aerobic glycolysis cause cancer.
Sydney Weinburg was correct in saying that mitochondrial defects, including
respiration were not the cause of cancer, but he was wrong in stating that aerobic
glycolisis is not required for cancer. Bambeck was the most correct of all in stating

30
that cancer is CG driven in a way to institute a glycolytic/mitochondrial differential
substrate and ATP flow state that is characterized by rapid glycolysis and a
mitochondrial respiratory/OX/PHOS shortfall, on a per cell basis. He felt this to be
true, regardless of the cause of the shortfall, be it an actual mitochondrial defect, a
paucity of mitochondria or a mitochondrial control element flaw. There was no
knowledge of a TOR, PGC-1alpha, 4E-BP mitochondrial regenesis/neogenesis
control triad or a CR pathway at that time, but the relationship between glycolysis,
the mitochondrion, and the substrate and ATP flow was there, and plainly visible, all
the same. We present Figure 2, reprinted, exactly as it appeared in his 1981
dissertation, as an example of his work and overall understanding at that time [27].

From our discussion of the relationship between glycolysis and the mitochondrion,
we conclude that it is the stuck state of aerobic glycolysis that remains as one of
the chief and necessary hallmarks of the disease, and in fact, that it is this stuck
state that is a cause of cancer, and that this state is a necessary and important one,
because it sets the stage for all the, most probably ROS induced, mutationally
aggressive changes that must follow to implement lethality; i.e., the avoidance of
immune surveillance, tissue invasion via mesenchymal conversion and metastasis
being the most deadly alterations required. Later alterations in the rate of aerobic
glycolysis, are also deadly because of its acceleratory impacts upon cell growth and
division speed. The fact that the system can be successfully attacked at this
metabolic level, that the rapid uptake of 18-F-2deoxyglucose by tumors is the
number one diagnostic tool for cancerous tumors and their metastatic detection, and
the fact that stuck aerobic glycolysis is a cancer cell growth requirement, pretty
much cements the cancer cell metabolism case shut. Primary defects at the
headwaters of the CG and CR systems appear to be the most likely principal causes
of cancer, but they funnel into a metabolic machinery that is therapeutically
sensitive to assault. We request that all readers note here, that the cancer portion of
this paper is devoted to the metabolic bioenergetic and substrate flow alterations
which are necessary to the maintenance of the cancerous state, even though we
discuss other elements of cancer that are related to these alterations. We are now
prepared to move on to the CG and CR disease manifestations outside the cell.

31
Figure 2. Substrate and ATP flow alterations in the cancer cell, as presented in
the 1981 Bambeck dissertation at Kent State University [27].

32
4) Diabetes, Metabolic Syndrome and the Map

Diabetes has been called a disease of starvation in the midst of plenty. It disrupts
the carbohydrate balance of the whole cell population of the body. It does so by
coupling massive carbohydrate intake, over a long time, with a muscular lassitude
that fails to burn the carbohydrate fuel and creates a catabolic lipid oxidizing
collapse, a pre-diabetic metabolic condition called metabolic syndrome (MS), and
also sometimes referred to as syndrome X, that not only maintains, but exacerbates
the condition. Thus, in the preponderance of cases, diabetes is preceded by obesity,
as the popular press never lets us forget. We now know that obesity exacerbates the
development of both cancer and diabetes II through the common pre-disease
pathway of elevated circulating insulin (I) [4]. The overall physiological response is
rather slow to develop, but has recently been revved up in the population by a
childhood obesity epidemic. The MS state is both a hyperglycemic and
hyperinsulinemic state, before eventually developing into full blown diabetic I
deficiency due to exhaustive pancreatic collapse in I producing capability. Simply
put, at the diabetic conversion from end stage MS, glucose is in abundance, and the
cells cant take it up. Many now feel that diabetes progresses from a disorder of
liver/muscle/adipose origin. These tissues occupy more than half of the
metabolizing wet tissue weight of the body in normal, non-obese individuals, and
thus, is a disease process facilitating bulk flow mismanagement of the whole
carbohydrate/lipid economy. This is now thought to be due, in part, to a progressive
obesity caused fatty liver induction of a downfield tissue ppar gamma deficit that
aids in the inhibition of both muscle fatty acid metabolism and the movement of
fatty acids from the adipose compartment to the muscle compartment. In the end,
elevated glucose, increased I resistance and I insufficiency cause glucose starvation
in I requiring cells, in addition to hyperglycation, elevated ROS, widespread
inflammation, and with a massively elevated cancer incidence and cardiovascular
system collapse risk, instituting a total body rebellion that disrupts virtually all of
the physiological mechanisms of the whole body. There are I independent tissue
cells in the body, like in heart, brain and breast, but they too, suffer from the ravages
of ROS, hyperglycation and inflammation. We describe more of the physiochemical
relationships, below.

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Early in the twentieth century, type I diabetes was discovered to result from loss of
pancreatic insulin production, which is now known to result from an autoimmune
destruction of pancreatic beta islet I producing cells. I replacement dramatically
reduced lethality, albeit with an over all reduced life expectancy. Middle age onset
type II diabetes also responded to I replacement, and was most pronounced in the
obese. Thus, initial research focused around a food intake (intestinal), liver and
pancreas metabolic triad. In the late twentieth century, the growing obesity
epidemic, the growing fatty liver (steatosis) epidemic, increased muscular lassitude,
a massive upsurge in fructose in the diet and key inter-tissue lipid transport
discoveries, have shifted our focus toward a muscle, adipose and liver triad, with
the pancreas, as mostly, a progressively failing response element to the triad.

The metabolic syndrome (MS), in modern developed nations, begins from a

combination of over indulgence of prepared convenience and junk foods, coupled
with a physically lethargic life style. These foods contain astronomical amounts of
rapidly absorbed simple carbohydrates, because these carbohydrates are almost dirt
cheap on a $ to weight ratio, making great filler and they also foster an eternal
sweet tooth addiction in humans, as described below. There are three main rapid
absorption carbohydrates in these foods, and they are: 1) high fructose corn syrup
(Hi FCS), which is composed of the simple sugar monomers fructose and glucose in
a 55/45 ratio, 2) sucrose, mostly from sugar cane, is a sugar dimer composed of
fructose and glucose in a 50/50 ratio and 3) simple starch, a mostly linear sugar
polymer composed of nearly 100% glucose. Hi FCS, being monomeric, enters the
blood stream fastest of all, achieving bulk transfer concentrations in about seven to
ten minutes, while sucrose must be cleaved into its monomers by the enzyme
sucrase, to also achieve similar to Hi FCS blood levels, in about twenty minutes.
Simple starch, as found in potatoes, rice, white flour etc., is partially cleaved into
large chunks by salivary amylase, then, converted to glucose only monomers by
further amylase treatment in the digestive tract. It is bulk transferred to the blood
over about an hour. Although all three are rapidly absorbed, Hi FCS is, by far, the
worst offender, due to its prodigious and rapid uptake, so we will use it to
demonstrate our MS to diabetes II conversion. Hyperglycemia is the cause of
protein glycation, and a previous month long diabetic life style management of
glucose can be measured as glycated blood hemoglobin, known as HbA1c.
Bambeck first became involved in diabetic research by developing an

34
electrophoretic isoelectric focusing glycosylated hemoglobin (HbA1c) diagnostic
test for Isolab Inc. in 1981. The test also detected sickle cell anemia, fetal
hemoglobinopathy and 70 other mutational hemoglobinopathies. The test gained
FDA approval and was widely used on newborns, mostly for fetal and sickle cell
hemoglobinopathies, throughout the 1980s and 90s. In diabetics, elevated glucose
damages protein function all over the body by reacting with them via glycation, just
as it does with hemoglobin. Glycation, and the resulting inflammation, is a major
component in the later development of diabetic complications. For the above
mentioned reasons, glucose management is critical, in diabetics.

When the Hi FCS monomers rapidly enter the blood stream in very high
concentrations, the sharp glucose hyperglycemic state initiates a rapid I spike, that
causes intense cellular uptake of glucose, soon instituting a precipitous glucose drop
in the blood, manifest as hypoglycemia. I over response, in the first place, results in
an unusual circumstance, not common prior to the ready availability of Hi FCS
foodstuffs, manifest as hyperinsulinemia co-incident with hypoglycemia. This
condition initiates a huge craving for sweet tasting stuff, to relieve the conflict
between the apparent fuel need and the I induced over response as a trans plasma
membrane fuel driver. This creates a vicious cycle of eating and craving, that fosters
the obese condition. This is the pre-MS condition. The second, and more covert
sugar, is fructose. Fructose in the blood stream does not induce insulin secretion in
the vast majority of humans, and is not readily taken up by most somatic cells.
Instead, it is sequestered in the liver to a high extent, where it is converted to triacyl
glycerols, which are then, esterified to fatty acids to form triglycerides, which are
stored in the liver as fat. Over time, this causes fatty liver (steatosis). Steatosis up
regulates Apo B protein, causing increased production of the system wide delivery
protein, called very low density lipoprotein (VLDL), and thus, increased
triglyceride transport to, especially, adipose tissue. Elevated VLDL is also
implicated in cardiovascular disease via the generation of arterial plaque, as
discussed, later. Chronic steatosis, if left unchecked, can institute early stage liver
cirrhosis in about a decade or two, then relentlessly progresses to advanced liver
cirrhosis and death, or to advanced liver cirrhosis, hepatic cancer and death,
depending on the luck of the draw. Progressive liver failure causes increasing
hepatic ROS, pro-inflammatory molecule production and increasing loss of
detoxification functions. Inflammation inducers and toxins flood the vascular tree,

35
and initiate system wide molecular damage that induces peripheral somatic cell
ROS and inflammation throughout the body, and a vicious cycle ensues,
progressively worsening the conditions. A well published wide range of disabilities
arising from this disease is available to all, from readily available popular books, to
uncountable sources on the World Wide Web.

It is estimated that thirty percent of the American population suffers from liver
steatosis, that the disease is appearing in large numbers, in obese teenagers, for the
first time in history, is occurring in parallel with diabetes II, and is a harbinger of a
need for future liver transplants in the untold thousands, or even millions. (A more
thorough review of fatty liver disease can be found by J.C. Cohen, et.al., Science,
vol. 332, p.1519, 6/24/11) [28]. Does this mean that we add a liver transplant
epidemic to the already existing obesity, steatosis and diabetes II epidemics? In
addition, what do we do about the fact that these traditionally middle and old age
onset conditions/diseases are now commonly appearing in young adults?

Silibinin, one of our phytonutrient gold medal winners, was originally used to save
mushroom poisoning victims from hepatotoxic liver failure. Later, it was put into
use for general hepatotoxicity. Medical drug applications, like some actors, get
type cast for their function and more creative research becomes stalled, as it did
so, for silibinin. Just recently however, silibinin has also been found to be useful as
an adjuvant to conventional cancer therapy across numerous tumor types, thus
showing a metabolic commonality between cancer and diabetes. We would note
here, that anyone thinking of linking silibinin along with the modern sugar junkie
diet would be living in a fools paradise. Silibinin is now known to act near to, or at
AMPK to initiate the CR pathway activation cascade, and to thus, relieve the pre-
diabetic and some of the diabetic conditions, just as does metformin. In fact all of
our gold medal finalist food supplements are CR mimetics, and perform this same
function, but with varying degrees of bioavailability. Now, we can look at some of
the diabetic muscle and adipose interactions, and their CR connection.

Inactive muscle does not raid adipose tissue for fat energy because it is white fiber
muscle, is almost totally anaerobic and burns glucose to lactate, for export to the
liver for gluconeogenic reconversion to glucose and then export back to the muscle
as fuel, the whole process known as the Cori cycle. Highly athletic endurance red

36
muscle fibers are the ones that use fat and oxygen as fuel for the Krebs cycle.
Couch potatoes have virtually no red muscle. It has recently been found that white
muscle is almost absent of peroxisome proliferator activated receptor (PPAR
gamma), a vital component necessary for the transfer of fat from the adipose
compartment to the muscle compartment. Not only is it true that white muscle will
not oxidize fat, but it cannot. Red muscle makes PPAR gamma, and both, will and
can oxidize fat. If there is enough muscle tissue oxygen demand, over several
months of intense endurance exercise used in concert with CR activators, white
muscle content can decrease and red muscle content can increase, and red muscle
mitochondrial content can rise from a lowly 5% of muscle volume to over 30% of
muscle volume, most probably as a result of the SIRT 1 CG subroutine described
earlier. However, that is not the case for the sugar sucking and lethargic humans we
have been discussing so far. At this point, sadly, their condition has become full-
blown MS. Lastly, tissues other than muscle respond to MS hyperinsulinemia and
hyperglycemia by down regulating their insulin receptors, while muscle up
regulates its insulin independent Glut 4 receptors; the same receptors found in
insulin independent cancers.

It becomes obvious why whirlwind exercise programs and crash diets will not work
on a slowly developing set of circumstances, like those outlined above. If not
checked, the liver/adipose/muscle triad system simply wears out the pancreas with
its chronic I requirements and the organ can no longer provide the demanded I: this
then is diabetes II, and without externally supplied I, the victim will die. From a
metabolic standpoint, it resembles a kind of CG/CM/aging and CR/HE/LE cellular
analogue that has gone awry at the tissue and organ level of integration. The
downstream activations are similar in both diabetes and atherosclerosis, in that
ROS, glycation and inflammation cause general tissue damage throughout the body.
It should be no surprise then, that diabetes dramatically accelerates aging, heart
disease and cancer incidence. With sugar intake reduction, moderate exercise and
our phytonutrient metabolic pathway modifiers discussed in the phytonutrient
candidate section, the preconditions can be halted and, to some degree, reversed.
CR mimetics can be very useful, here, because of their ability to reduce both
glucose demand and circulating I concentration, as shown in our metabolic pathway
map. The full blown disease cannot be totally reversed, but its progress can be
diminished or slowed, and we mean slowed for a long time, and by a lot.

37
Pharmaceutical metformin, a direct activator of AMPK, and thus CR/HE/LE, has
been widely used for decades in beyond MS low level diabetes II, to control blood
sugar and to delay insulin dependence. It decreases insulin resistance, up regulates
the CR/LE pathway and down regulates the CG/CM/aging pathway, and with all
our metabolic maps appropriate myriad downstream impacts. In a multi-patient
survival record analysis, it was shown to dramatically increase the survival time of
diabetic brain glioblastoma victims over those patients not taking metformin,
regardless of all other therapies used. Our five gold medal winners have a
mechanism of action similar to metformin, and they have all shown efficacy in
glucose control and across a startlingly broad array of tumor types [26, 27, 30].To
our knowledge, metformin has not yet been tested across a broad array of tumors,
but such research has already begun, in humans, and with respectable preliminary
results. Again, and this time with a pharmaceutical and CG/CM/aging and
CR/HE/LE rationale, cancer and diabetes show therapeutic linkage via a metabolic
commonality.

The metabolic syndrome/diabetes sequence can daisy chain into very serious
cardiovascular damage, as is both common knowledge, and as we shall see
presented in the next section. Of the three aging disease states outlined here,
atherogenic cardiovascular disease is the most cryptic and obscure, from the
standpoint of our CG/CM/aging and CR/HE/LE metabolic map, as the disease is a
mixed bag, of as much of an effect from other conditions, as it is from its own
causes. However, the disease responds, especially during its early and mid stages,
magnificently to CR mimetics.

5) Atherogenic Cardiovascular, Related Disease States and the Map

The rough and tumble environment of the vascular tree is far different than that of
the calm extracellular interstitial fluid backwaters bathing most of the bodys cells.
The vessel walls are directly exposed to mechanical and chemical stresses that arise
from distant sites and are delivered or manifest throughout the system. For instance,
hypertension accelerates the rate of formation of atheratomous lesions, initiating a
call out for low density lipoprotein (LDL) repair mechanisms. Also, MS, initiated
elsewhere, as hyperinsulinemia, hyperglycemia, hyperglycation and

38
hyperinflammatory ROS, TNF, COXII, ILs etc., cause molecular damage within
and throughout the lumen exterior and interior walls of the arterial system. The lipid
delivery system is somewhat unique to the circulatory system and has its own
peculiar ways of going awry, as we shall see, with LDL, HDL, foam cells and
arterial intima smooth muscle cells, in addition to the liver steatosis activation of
VLDL that we discussed in the diabetes section. Atherosclerosis is mostly restricted
to the much higher pressured and most thickly walled arterial side of the vascular
network, where turbulence inflicted damage is most pronounced, and where the
damage resulting from the reaction to the initial damage, is even more pronounced.
The response and response to a response sequence, somewhat obscures a direct
interpretation of our metabolic map. Rational salvation lies in the fact that the
disease causes and therapeutic outcomes share that same, previously mentioned,
Occams razor.

The classical low density lipoprotein (LDL i.e., bad cholesterol) and high density
lipoprotein (HDL i.e., good cholesterol) story is the most widely known,
medically instructed and publicly disseminated narrative in the history of medicine,
and will only be given a cursory brush over, herein. We will, however, give some
space to a few very lethal, but less known variations of the theme. The lipoproteins
LDL and HDL precursors are synthesized in the liver as a hyper-lipid packed very
low density lipoprotein (VLDL) and the lipid void form of HDL. VLDL and LDL
engage in lipid delivery to the peripheral cells via the vascular network, while HDL
acts as a lipid scavenger, return to liver, lipoprotein LDL and cellular transfer agent.
Hyperlipidemic disease participates in the cause of atherogenesis by most typically
manifesting as a genetic or environmentally induced over representation of LDL, or
less typically, as a genetic or environmentally induced under representation of HDL.
Thus, the notion of the LDL/HDL ratio, as the famous bad cholesterol to good
cholesterol index of cardiovascular disease risk, which has now become a common
household phrase. Environmental and genetic factors synergize, so that one born
with a genetic proclivity for atherosclerosis requires less of an environmental insult
than one born with a more normal genotype, as is also true for a significant fraction
of environmentally induced obesity, diabetes II and cancer victims. We try to
integrate our approach, so as to provide as few examples as possible.

In the normal process, VLDL in the blood stream slips between the endothelial

39
lining cells of arteries, veins and capillaries, where it is converted sequentially from
VLDL to intermediate IDL to LDL and then to a mostly lipid depleted LDL by
sequentially binding to B, E and B plus E receptors on cells throughout the body,
where lipids are off-loaded. The mostly lipid depleted LDL returns to the blood to
be decommissioned by the liver and replaced by nascent LDL. An LDL molecule
can be ROS oxidized, and becomes a B and E receptor independent lipid delivery
molecule to arterial wall intima layer residing macrophages sporting LDL lipid
scavenger receptors. LDL is all too frequently present in the population as a small
dense (SDLDL). The SDLDL form is much more readily oxidized, is viciously
atherogenic and has a very high affinity for macrophage LDL lipid scavenger
receptors. This converts the macrophages into foam cells, which will be briefly
described, later.

Meanwhile, much is happening on the HDL side of the equation. After being
converted to its active form, the lipid void HDL-3 mostly fills up with lipid by
binding to E receptors, which further activates a blood born new form, now called
HDL-2a to utilize the cholesterol ester transfer protein (CETP) for LDL interactive
transfer. The fully lipid loaded HDL, called HDL-2b, is captured by liver E
receptors and is decommissioned. There appears that some people have an unknown
defect that interferes with HDL-3 conversion to HDL-2a, possibly interfering with
Apo AII removal or Apo E activation. This causes HDL-3 to pile up in the blood.
Bambeck created a lipoprotein subractionation assay, over a decade ago, for both
SDLDL quantitation and HDL-3 quantitation, and in the process, became a little
more knowledgeable about their relationship to cardiovascular disease. His test also
revealed that there are, at least, two VLDLs, three IDLs, seven LDLs in two
diagnostically meaningful functional groups and thirteen HDLs in three
diagnostically meaningful functional groups. Except for a few intrepid individuals,
this lipoproteomic assay had a few too many components for the research and
diagnostic communities to digest at that time.

The quantitation of SDLDL should be a medical priority, as being atherogenic it

appears in about 27% of the general population,and does not appear on any
traditional cholesterol screen nor responds to the standard regimen of statin drugs,
but does respond to fibroates, such as gemfibrozil. Approximately half of all
SDLDL victims show no other risk factors with conventional diagnostics. About

40
eight or nine years ago, as contract work, Bambeck proposed that SDLDL disease
is represented by a single co-dominant allele of an unknown gene, the only other
allele being the normal form. The disease was proposed to manifest in its
homozygous form at about age twenty five and kill by age 45 in about 2% of the
population. More disheartening, is that in its hypothesized heterozygous form, it
manifests in about 25% of the population at age 45 and kills by age 65. The disease
is almost a perfect match for a two allele system in Hardy-Weinberg equilibrium
with a 14% general population penetrance. How it gained such penetrance remained
a mystery until it was discovered that grossly obese children of parents with
SDLDL, who were put on emergency lipid free diets, turned on the SDLDL
response at the unheard of early age of ten. That, plus the dietary response to
intestinally active gemfibrozil, indicated that the mutation had a strong gut lipid
extraction survival advantage in a small human race founder population that faced
severe nutritional lipid semi-starvation for many generations. Do not know if this
notion ever gained any traction. Regardless, SDLDL is a major league population
killer, for which resolution is readily available, if we would only test for it and treat
it. For now, abundant exercise, a low sugar and fat diet and CR mimetics to aid in
low ROS generation is all that exists for those poor souls who dont know that they
have SDLDL disease or cant readily locate a test to diagnose it.

The non-functional HDL-3 situation is less severe, but just as silent as SDLDL,
and appears in about one-fourth of the general population. About 90% of
individuals with elevated HDL-3 have an elevated LDL, but conventional
diagnostic assays report only the total HDL number, which includes HDL-3 in the
aggregate, thus under reporting the risk as a lower functional LDL/HDL than it
should be. The risk is even more skewed in the 10% without other known risk
factors. The impact of HDL-3 was not clearly defined until recently, and the
Cleveland Clinic bemoans the fact that there is no available non-functional HDL-3
test. Unfortunately, the source of the HDL-3 assay filed for dissolution in 2004. All
samples used for the lipoprotein subfraction assay came from 4,000 participants in
the Framingham Study, and were kindly provided by Judy McNamara of Tufts
University.

The lipoproteins are only part of the atherogenesis story, although, for most arterial
disease, LDL and its variants are most often considered to be the culprits. In

41
response to arterial wall injury due to baromechanical, chemotoxic or
inflammotoxic origin, macrophages migrate to the intima layer of the artery and
acquire lipids from LDL for use as fuel, repair and maintenance. In the abnormal
condition of hyperlipidemic LDL, high numbers of ROS induced OXLDL and/or
genetic SDLDL, macrophages become lipid overloaded, taking on a golden lipid
droplet engorged state, microscopically referred to as foam cells. Foam cells
hyperactivate an immediate vicinity NF-kB cascade and somehow induce mitosis in
the smooth muscle cells lining the intima layer. Necrotic foam cells make up much
of the fatty part of the forming atherosclerotic plaque, while replicating smooth
muscle cells form beneath a fibrous plaque capsid that acts as a surface seal to the
growing atheratoma, which eventually protrudes into the arterial lumen. Some
scientists have speculated that smooth muscle cell proliferation is akin to benign
tumor growth and that the macrophage foam cell and smooth muscle cell
inflammatory/mitotic cascade form a progressive self-reinforcing vicious cycle that
facilitates and exacerbates plaque growth and its spread as colonial groups. This
makes sense from a molecular damage stand point, and is consistent with the
CG/CM/aging pathway dynamic.

Ultimately, plaque artery blockage becomes, so severe, as to cause downstream

tissue ischemia and hypoxia. Sudden death often occurs when a plaque capsid tears
and fibrinogenic polymerization blocks the artery with a clot, or a clot breaks off
and lodges further downstream, causing anoxic death to vital function tissues such
as heart, lung or brain.

There are a host of other cardiovascular disease states, more particular to the heart,
that result from a shift from a CR/HE/LE pathway driven to CG/CM/aging pathway
driven status. Inflammation, ROS and glycation are neuropathic and can disrupt the
cardiac cycle, leading to arrhythmias, fibrillation and infarction, as well as causing
valvular and endocardial lining damage. All of the above can lead to a heart attack
or congestive heart failure. Mitochondrial inefficiency and low mitochondrial
numbers can reduce ATP production and force of contraction, as found in
congestive heart failure via ventricular hypertrophy. Shifting the drive state from
CG/CM/aging to CR/HE/LE has been shown to be neuroprotective, as well as
neuroregenerative in both heart disease and diabetic neuropathy. This metabolic
shift has been shown to increase mitochondrial numbers in both cardiac and skeletal

42
muscle, to increase ATP production and to increase muscular force of contraction,
thus relieving all the above mentioned conditions [25, 26, 29]. Our five gold medal
winning food supplement molecules perform all these functions, in addition to their
anti-diabetic and anti-cancer functions, as described earlier. The silver medalists
synergistically support the gold medalists. Finally, we are ready to discuss our
selected dietary supplements.

6) Food Supplements/Phytonutrient/Nutriceutical Candidates

The health food stores and giant pharmacy chains are awash with a bewildering
array of hundreds, if not thousands of natural (and otherwise) dietary food
supplements and herbal extracts from the four corners of the world. Just walking
down the aisles should institute some form of clinical depression. Where to begin?
How does one separate the questionable legitimacies of folk remedies, old wives
tales, heap big mojo shamanism, venerable ancient medical wisdom, mythology,
cultural favorites, modern marketing snake oil hucksterism etc. from actual fact and
good medicine? As an alternative to taxing our already enfeebled brains, we hit on
the best idea weve had in years. Instead of embarking on an interminable grand
search, we let the worlds best and brightest topic specific health and life extension
professional brains do our thinking for us, and we went in with a strategic plan.

We embraced a copy Disease Prevention and Treatment, Fourth Edition, published

by The Life Extension Foundation (LEF) by Life Extension Media, cancer: pp. 209-
406, cardiovascular disease: pp. 413-521, diabetes II: pp. 709-742, obesity:
pp.1241-1272, 2003 [29]. Therein lies the combined efforts of thousands of
research studies and the clinical experience of physicians around the world (to put
it in LEFs own words) devoted to the over riding purpose of LE. After reviewing
what food supplement molecules have and yield, the proper CG and CR peripheral
inputs and outputs, respectively (based upon our map in Fig. 1), and a history of
appropriate medical outcomes, we used scientific publications and the Internet to
update and modernize our understanding, mostly in hopes of finding the food
supplements most active ingredients molecular targets and/or mechanism of action
[25, 26, 29]. Having our CG/CM/aging and CR/HE/LE map in hand, and our three
principal diseases to both direct our search and institute the strategic plan, we
plunged in.

43
The plan had a few simple but inviolate and all inclusive rules. Each candidate must
have utility against multiple cancers and against diabetes and against
atherosclerogenesis; all three It must both statistically prevent or delay onset of all
three diseases by significantly reducing the probability of age related population
incidence, and slow the progression of all three disease states, once disease is
instituted. Where the principle target and/or mechanisms of action are known, it
must regulate its target, and in turn, their downstream pathway targets in the
direction consistent with the CG/CM/aging and CR/HE/LE map. Where principle
target or mechanism of action is not yet known, downstream effects must match the
regulatory directions of our metabolic map. Although not an absolute requirement,
each chemical should have an extremely high LD-50, in the multiple dozens to
hundreds of times its functional dynamic range and a history of extreme safety. All
our winners achieve this. The probability of satisfying all of these and
requirements is minute, and a powerful set for a true candidate.

Limiting our search from the world at large to the LEF compendium, instantly
whittled the search from thousands of possibilities to a few hundred. Applying the
all inclusive rule set to these few hundred, fairly quickly drove the number down to
a few dozen and then, much more slowly and agonizingly, chipped its way down to
a handful. Incidentally, route of administration was not a consideration as some of
these molecules have poor oral bioavailability in a pure form, but much higher
bioavailability in a soluble injectable form, or in a conjugated oral form. Winning
candidates might be more accurately defined as nutriceuticals rather than as
phytonutrients or dietary supplements due to their mechanism of action. We have
already provided a general description of the differential therapeutic merits of our
gold medal and silver medal winners, and we will diversify those merits and
differentials, later.

Before we begin, we must say a word or two, about antioxidants. For one thing,
antioxidants are good for us, plain and simple. Thus, it is not surprising that
antioxidants have been all the rage for the last two decades, and for a host of good
reasons. They protect us from the major diseases of aging, reduce ROS formation,
give us more vitality and with a long list of well worn etceteras, allow the
population survival curve to leptokurtically shift toward the maximum normal life

44
expectancy. However, they simply do not institute LE, albeit having been, up to this
point, the best game in town. Driving this point home is that organisms on a CR or
CR mimetic protocol manifest LE without any antioxidant supplementation
whatsoever! If this were chess, that alone would be check and mate against
antioxidants as LE activators.

For this and other reasons the antioxidant impact on the ROS damaging hypothesis
is now seen more from an HE and avoidance of the life shortening side, as opposed
to the life lengthening side of the equation. Thus, the antioxidant hypothesis is
found to be limited and in need of modification. We must remember here, that the
most critical elements of the CR/HE/LE pathway were only discovered during the
last two to three years, so a theory modification was not even realistically possible
prior the very recent present. We must remember also, that researchers prior to 2005
had no real knowledge of anything like CR metabolism regulatory controllers or
that CR metabolism could be mimicked by either pharmaceuticals or extracted food
supplement molecules. For instance, from an antioxidant standpoint, the CR/HE/LE
pathway institutes mitochondrial regenesis and reduces ROS production, in the first
place, obviating the need to mop up ROS with antioxidants. Prior to the elucidation
of the CR/HE/LE pathway, any LE system activator would have been, and in fact
was called an antioxidant, simply because mitochondrial neogenesis and regenesis
were not separated from biogenesis at the time. Even earlier in the twentieth century
and before, the sources of these molecules were relegated to herbal remedies and
folk medicine as liver, blood, digestive and whole body purifiers. We believe
ourselves to be the first folks to attempt to formulate a primitive global
CG/CM/aging and CR/HE/LE map that includes the notions of cancer metabotype,
atherosclerogenesis, diabetes, mitochondrial neogenesis/regenesis, CR/HE/LE
pathway, CG/CM/aging pathway metabolism, CR/HE/LE pathway mimetic and
functional antioxidant classification as simple antioxidant, funnel antioxidant and
metabolic pathway antioxidant, even though all of these things are well described in
the literature, but disjointed, because this age of specialization obscures the big
picture overview approach, such as ours. We know of no scientists that can talk
fluently across such a multiplicity of topics. We cant either, but it all becomes
rather simple and straightforward when one has the map: and that is the beauty of
this whole endeavor.

45
As we said before, the health food supplement world is awash in so called
phytonutrient antioxidants, and our metabolic map has helped us to loosely classify
them into three broad categories with considerable overlap, principally because of
the importance of CR in activating the cells anti-ROS mitochondrial regenesis
program. Simple antioxidants are, to put it simply, basically just antioxidants.
Vitamin C is a simple antioxidant. In fact, if it doesnt have a partner to defuse it, it
becomes a pro-oxidant in its ROS activated state. Funnel antioxidants are
antioxidants that both accept ROS from other antioxidants and pass their ROS
activated electron(s) to a metabolic system to defuse them and to extract useful
energy. For instance, the vitamin C, vitamin E, NAD sequence, sets vitamin E as a
funnel antioxidant. Better yet, a little system like coenzyme Q (CoQ) and alpha
lipoic acid funnel ROS energy from ROS and many other antioxidants, with CoQ
being a requisite functional element in the metabolic pathway itself, for passing
ROS electrons to the OX/PHOS system of mitochondria for energy capture as ATP
production. In addition, alpha lipoic acid restores vitamin E and vitamin C to full
antioxidant status by reducing their oxidized state; a possible recycling alternative
to massive simple antioxidant dosing. Best of all, are giant macromolecular
machine system antioxidants such as the mitochondrial OX/PHOS efficiency
system of mitochondrial regenesis, as turned on by activated CR/HE/LE. This third
category is the big antioxidant player in our new understanding of the CR/HE/LE
mitochondrial neogenesis vs. regenesis distinction. Now, we are in a position to
consider the functions of our winning candidates. Sadly, we cannot list all of the
impacts of our gold and silver medal winners, or this narrative would read like a
phone book. We will bundle wherever possible and stick to main effects, hopefully,
without short changing the reader.

The five big gold medal winners are curcumin, silibinin, berberine, resveratrol
and pterostilbene. Cumin root can be partially purified to turmeric, taken as is, or
further purified to its most active ingredient, curcumin. Milk thistle extract, called
silymarin, can also be taken as is, or can be purified to its most active component,
silibinin. Berberine is most often provided as an extract of the popular herb
goldenseal. Resveratrol is most often alcohol extracted from waste grape skins or
Japanese knot weed roots, and provided as an impure mixture of about 80/20 to
60/40 ratio of phytosterols to resveratrol and consumed as is. It is also available in
its most active form, trans-resveratrol. Pterostilbene is a dimethylated derivation of

46
resveratrol, and is found in vanishingly small quantities in purple berries, with the
blueberry being the source of its fame. Except for pterostilbene, all are cheap to
extract from plants, are readily available, have a long historical tradition, own a
virtually immaculate safety record, are un-patentable and are causing
pharmaceutical companies to rip their hairs out by the roots, in exasperation. On the
pharmaceutical upside, the companies are assiduously pursuing better, more
biochemically focused, patentable, and most importantly, more expensive
pharmaceutical alternative mechanism of action variants. But, until then, we are
stuck with the natural stuff. Because their overall impacts and principal active sites
are so similar, as described below, we bundle all five together. Of course, as
downstream mitochondrial regenesis activators, all have been historically listed as
powerful antioxidants, but now we know better.

As succinctly as possible, all five gold medalists down regulate AKT, COX II,
inflammatory IL cytokines, NF-kB, HIF, VEGF, angiogenesis, fasting insulin,
insulin resistance, LDL, blood glucose, HbA1c, ROS, all incidence and rate phases
of cancer cell initiation, progression, proliferation and invasion, as well as all the
phases in the obesity, liver steatosis/cirrhosis/MS/diabetes sequence and the
atherogenic cardiovascular and related disease processes and groups, to name a few.
In addition, they all up regulate apoptosis, autophagy, insulin sensitivity, AMPK,
P53, insulin dependent receptors, heart and skeletal muscle oxygen consumption,
ATP output, mitochondrial numbers, force of contraction, hepatogenesis,
neurogenesis, mitochondrial regenesis, antioxidant response and multiple type
cancer cell differentiation and disease victim survival time, also, to name a few[25,
26, 29]. These data imply that that they must be operating upstream of TOR, close
to a main input branch of TOR, perhaps on the CR side not far from AMPK, due to
strong AMPK and p53 system up regulation in the absence of evidence supporting a
very strong AKT, P13K, or RAS down regulation. A little over a year ago,
Affymetrix gene chip shotgun analysis showed that resveratrol activated exactly the
same near 1,000 gene set activated by CR downstream of AMPK in all tissues
tested, those being muscle, brain and liver. We learned, after our map had predicted
it, that curcumin activated the same gene set and that pterostilbene turns out to be a
much more bioavailable (about 30X) analogue of resveratrol. Berberine also
activates all the same downstream pathways activated by AMPK.

47
We suspect a similar outcome for silibinin, as it has been recently found to directly,
or close to directly, up regulate P53 and its down stream cancer suppressor protein
P21. The P53 interactions with AMPK may be complex and self reinforcing,
because P53 is up regulated in all five of the CR mimetic cases, which seemingly,
should not occur because one would expect eventual cyclic feedback inhibition, as a
result of mitochondrial regenesis, from a simple or direct AMPK activation.
However, this same result is achieved from use of the well characterized
pharmaceutical AMPK activator, metformin. The rather soft deactivation of the
CG/CM/aging pathway with all five of the food supplement CR mimetics shows
itself to be due to cross regulatory pathway inhibition by CR pathway activated
factors such as P53, PTEN, P21 and others, causing partial TOR block and
diminishment of the mitochondrial RTG response, rather than due to direct
upstream CG pathway inhibitors, which would still be in operation because of
plentiful food availability, if one is only doing CR mimetics, and not dieting.
Fortunately, CR mimetics reduce circulating glucose and turn on fat burning in its
stead, which helps to reduce the sugar junkie appetite and to counter obesity.

Oral bioavailability of curcumin, silibinin and resveratrol is poor, in all cases. Oral
bioavailability of curcumin increases 2,000% when mixed with piperine from black
pepper. Silibinin is obtainable as bioavailable conjugates of several kinds.
Resveratrol is available as lozenges to support buccal absorption, or is provided in
pure form, in mega-doses. Resveratrol conjugation would not work orally, because
its unprotected hydroxyl groups are converted to kidney eliminable glucoronides
and sulfonates in the intestine and liver on the first blood pass through these organs,
which occurs prior to the first pass to the somatic tissues, resulting on only 2-3%
actual metabolic bioavailability to the cells. Pterostilbene seems to emerge from the
pack as being the strongest nature based CR mimetic contender. Its two resveratrol
analogue methylated hydroxyl groups block glucoridiration and sulfonation,
resulting in 64 to 91% bioavailability, depending on the experiment one consults.
This makes pterostilbene about thirty times more bioavailable than resveratrol.
Even though pterostilbene is only present in vanishing quantities in nature, its
simple structure renders it easy to synthesize from natural precursors.

The remaining candidates failed to make our rigorous gold medal cut for one reason
or another, but they are not slouches by any means, and that is why we give them

48
the high status of the words silver medal. The silver medalists are all excellent
supplements and form powerful synergies with each other and the other five big
gold medal winners. Unfortunately, all five gold medalists only exert their main
effect on the CR/HE/LE pathway, depending entirely on regulatory CR/HE/LE
elements to inhibit CG/CM/aging. Some of our silver medal winners are direct
CG/CM/aging down regulators, while others are CR/HE/LE reinforcers, and one is
even a shotgun multi-system synergizer.

The shotgun synergizer is a real powerhouse, a whole system appearing to have

been evolved to operate it. It is the omega 3/6 essential oil mix available from flax
seed oil, fish oil or krill oil, with flax being by far, the least expensive. The most
important are the omega 6 linoleic acid, the omega 3 gamma linolenic acid, and
their metabolites omega 3 eicosopentanoic acid (EPA) and 3 docohexanoic acid
(DHA). Flax seed oil is more than 60% omega 3 gamma linolenic, which is readily
converted by enzymatic processes, in the body, to EPA and DHA. These fatty acids
incorporate into cell membranes and give them durable integrity, reducing
brittleness and thus, increase all membrane dependent cell function efficiency. They
are metabolized to become a number of cell system activators and inhibitors. For
instance, the DHA and EPA pathway products reduce arachidonic acid, IL1, COX II
and TNF induced inflammation. Other impacts include increasing apoptosis and
immune surveillance, and reducing cancer cell proliferation, cachexic wasting and
angiogenesis. They act as anti-metastatic, anti-HIF, anti-VEGF and are essential for
mitochondrial cardiolipin production, which is critical in the efficient function of
the mitochondrial complexes III and IV in the respiratory chain [BIG THREE
REFERENCES]. The American diet is seriously deficient in omega 3/6; some say
suicidally deficient, containing less that 10% of the healthy minimum needed. Some
of the omega 6 oil is shunted down the proinflammatory arachidonic acid
production pathway, but if the anti to pro inflammatory metabolite producing
omega 3 to omega 6 ratio, respectively, is greater than 1.5 to 1, the overall response
is anti-inflammatory. For instance, the flax oil 3/6 ratio is about 3:1, far beneath the
tipping point to net inflammation.

Another synergistic beauty is alpha lipoic acid. The Linus Pauling institute
considers it to be one of the most powerful antioxidants known. Of all of the
winners listed, it would be classified as a zoonutrient, but is only actually available

49
as a synthetic, as natural source isolation is ridiculously expensive. Fortunately, its
structure is so dirt simple that its synthesis is also dirt cheap. After having
reacted with a free radical, it can be regenerated from its oxidized form by
glutathione. It is also a funnel anti-oxidant, regenerating vitamin C, vitamin E, the
ubiquinone form of CoQ and all the while, destroying a wide array of ROS. It
reduces NF-kB and foam cell production and renders regenic mitochondria more
efficient [26, 27, 30]. In fact, alpha lipoic acid preferentially locates to
mitochondria, where it has the most ROS reduction impact. This strongly supports
the CR/HE/LE side of the equation.

Coenzyme Q (CoQ) runs along similar lines as alpha lipoic acid but its primary
function is more specific. It is a powerful antioxidant in its own right as ubiquinone,
but preferentially locates to mitochondrial respiratory complex I, where it increases
net mitochondrial ATP production via enhanced electron transfer efficiency. CoQ,
along with lipoic acid, activate glutathione production in the liver and assist with
glutathione ROS reduction throughout the body. Glutathione is the most powerful
antioxidant produced by the body, is a key alpha lipoic acid regenerator and has
multiple impacts all over the CG/CM/aging and CR/HE/LE systems, particularly
with NADPH in the cytoplasm during CG driven neogenic induction of
mitochondrially elevated ROS production. CoQ has long been used in Japan to fight
congestive heart failure and left ventricular hypertrophy by increasing cardiac ATP
production and force of contraction. CoQ assists the CR/LE pathway in helping to
achieve its full capacity as mitochondrial regenic efficiency.

The most active ingredient in green and white tea is epigallo catechin gallate
(EGCG) while the most active component in soy extract is genistein. They both
inhibit the CG/CM/aging pathway somewhere downstream of the growth factors,
and upstream of AKT. There is some evidence that EGCG might operate
somewhere between growth factors and RAS. They both have broad spectrum
tumor anti-proliferative activity, and where measured, down regulate the
CG/CM/aging to TOR upstream activators. Another powerful CG inhibitor which
operates directly on AKT, is inositol 6 hexaphosphate (IP6) (described
previously), which can be extracted from rice grains. Finally, we have
CG/CM/aging down regulators to complement our five gold medal CR/HE/LE
activators! It seems to make sense, that if one were to take both a CG inhibitor and

50
a CR activator, the metabolic system should operate as if under much more realistic
CR mimicking conditions, because it should render food intake much less relevant.
With further research, maybe EGCG, IP6 and gensistein might someday be elevated
to our gold medal winner status. We still need to know whether CG inactivation can
act as a CR synergizer of LE.

Vitamin K, or more significantly vitamin K2, might someday be included as a

player on the CG/CM/aging side of the equation, as it is a tyrosine phosphatase
inhibitor of vital elements in the cell growth cascade. It has been very recently
discovered that vitamin K2 has the surprisingly unexpected function of operating in
the mitochondrial cytochrome chain as an electron donor that complements CoQ,
and that if it is absent, it might result in mitochondrial electron transport deficiency
diseases (S. Bhaleral and T. R. Clandinin, Science, vol. 336, p. 1040, 6/8/2012)
[30]. Thus, vitamin K2 may also act downstream of TOR as a CR/HE/LE pathway
synergist. There also might be a few more winners among the supplements already
in common use that simply have not yet been tested within the framework of the
CG/CM/aging or CR/HE/LE system. All in all, we conclude that alterations in
carbohydrate metabolism are responsible, to a large degree, for generating the great
diseases of aging, and that these diseases can be avoided, inhibited in development
or reduced in severity with the rational inclusion of specific metabolic function
defined food supplements.

7) An HE/LE Dietary Protocol for Even the Laziest of Minimalists

Practice the following regimen, if possible, daily.

1) One good multiple vitamin: particularly a brand that includes small amounts
of a wide array of metal cofactors, taken once a day with a meal.
2) A cell growth inhibitor and caloric restriction mimetic: from our food
supplement listing in the contents section 6, preferably EGCG (100 mg) and
pterostilbene (50 mg), once or twice daily, with food.
3) Omega 3 oil: from flax, borage, fish, krill etc. (1 tablespoonful, or 14 grams),
taken once a day with a meal.

51
 4) Dense salad: a fresh multiple vegetable salad that is either finely diced with a
food chopper or liquefied with a blender, (one 8 oz. cupful before the morning
meal and one 8 oz. cupful before the evening meal), as an appetizer).
5) Antioxidant/vitamin supplementation (optional): vitamin C (1,000 mg),
vitamin E, (400 I.U.), alpha lipoic acid (100 mg), coenzyme Q (50 mg) and
vitamin D3 (5,000 I.U.), all taken once daily with a meal.

The need for number 1), a good multiple vitamin with small amounts of a wide
array of metal cofactors has to do with the fact that farm monoculture, and in
particular, the use of insecticides and fungicides, although necessary on massive
farm scales, result in poor microbiota and detritivore mobilization of soil
micronutrients to plant roots, leaving our food deficient in many of these vital
cofactors. Coral calcium consists of a virtual periodic table of these necessary
micro-elements, and should be a reasonable alternative. Number 2), the CG
inhibitor and CR mimetic combination is designed to act synergistically, so as to
reduce both the obesity causing sugar/hunger drive state, and the incidence and
severity of the three great diseases of aging. Although we are not in the business of,
nor do we intend to engage in the practice of, product promotion, we have only
found one product which satisfies both of our CG and CR requirements. It is called
Eternal Blue, and it is manufactured by Blue Science. It came out on the market a
mere three months prior to the completion of this paper, and is the product that we
would have invented if we owned a $100 million food supplement processing plant.
Item number 3), omega 3 oil, is adequately described in our food supplement
candidate section.

To the truly lazy minimalist, our number 4) item, dense salad, sounds like a chore,
but there are some things that one may wish to consider (including the chore part).
If you dice up, or liquefy three bowls of a typical salad, you end up with only one
bowl of dense salad. This shows that three bowls of a standard restaurant tossed
salad (mostly lettuce) consists of two bowls of air, to yield a net of, one bowl of
actual food. Also, prepared foods have trained us not to chew properly, and poorly
chewed vegetables have very diminished digestive absorption. Dicing or liquefying
vegetables makes their metal cofactors and micronutrients much more bioavailable.

52
Liquifying the vegetables in a blender is very fast and easy. Throw in the juicy stuff,
(like tomatoes) first, then follow with a veritable cornucopia of day to day vegetable
options and taste variations, including nuts, berries and whatnot, to obtain
something similar to the consistency of a smoothie or a milk shake. If its
consistency is too thick, add a couple ounces of juice or water. This should be made
and consumed fresh, because many phytonutrients are lost via enzymatic, acid /base
and redox decomposition within a few hours. The beauty of the blender is that after
youre done making this delightful liquefaction, you can rinse out the reservoir, fill
it half full of tap water, add a few drops of dish detergent, set the machine on
blastomatic and it will blend itself clean. Rinse with tap water, and youre done.
This is true minimalism at its best, because you spend most of your time just
watching things go around in circles, and besides, it is also a practical answer for
veggie haters, who use the excuse that all that cutting just takes too much time.

The number 5) item, the antioxidant and vitamin supplement option, from the
antioxidant standpoint, is just a backup plan because the CR regimen turns on the
cells 200+ gene activated macromecular machine antioxidant systems anyway.
Coenzyme Q is rather useful to most people over the age of fifty because it has been
shown to strengthen force of contraction in the heart. Vitamin D3, the so called
sunshine vitamin, is probably the least optional, and most necessary, on the list. It is
important because it has untold numbers of functions and is in short supply polar of
the thirty to fortieth latitude lines, in both hemispheres, due to the lack of high
vertical angle sunshine, especially during the four deepest winter months. Vitamin
D is produced by the body in adequate quantities if the skin of the arms, legs and
head are exposed to high angle sunlight for only about 20-30 minutes per day. This
is not enough sunlight to burn even the palest of skin tones. For these reasons, we
should supplement during the colder months of the year. It is important to note that
the darker the natural skin color, the more one needs to supplement, due to the
blocking of D producing rays by melanin.

Of course, proper exercise and diet are a bonus, but for all those entrenched couch
potatoes and absolute minimalists out there, our dietary protocol is the best
alternative option we can think of. The protocol can be started at any age, and the
effects of CR/HE/LE pathway activation have been shown to have the most
pronounced effects on those of us who have more than fifty years under our belts.

53
8) Conclusion

We all know that there is no perfection in this real world, and we are far from
projecting perfection herein. Animal studies show that CR and CR mimetics induce
HE/LE, even when started during the pre-adult growth phase. They do not inhibit
normal mitosis or cell differentiation, but the resultant animals have slightly smaller
cells and body size. We must remember that CG is a good thing, in its proper time
and circumstance, for without it, we wouldnt even exist in the first place. With that
being said, it would seem to be prudent not to apply CR mimetics or CG inhibitors
when pregnant, still growing, taking intentional ROS generating cancer
chemotherapy, suffering from GH deficient dwarfism or being immune
compromised by such items as rapamycin, organ transplant or a large, late stage,
HIV load. Anyone going in for major surgery should stop CR, and in fact, should
probably begin a preparative GH therapeutic regimen at least five days prior to the
surgery, so as to promote wound healing. GH provides no benefit if taken after
surgery. Of course, you should always consult your doctor before trying nutritional/
dietary changes of any kind.

Not surprisingly, there have been a few internet blog inklings of the unsubstantiated
possibility of increasing the probability of such normally common conditions as
kidney casts and gall bladder/bile duct involvement. These would necessitate multi-
ten thousand people population studies to weed out a single cause from the land fill
of known entities, as 50 million Americans contract these conditions during their
lifetimes, anyway. Besides, no human studies have been conducted, nor have been
deemed worthy of being conducted, on these topics. Considering the vested interests
(i.e. no CR patents, no CR clinical pipeline) of some of the sources of these rumors,
and there being no real beyond arms length data, we chalk this up to purposeful
misdirection or typical internet scare tactic mythology. We must note however, that
there is a weak association between curcumin over use and biliary obstruction and
exacerbation of existing stomach ulcers if curcumin is taken, regularly, on an empty
stomach. The recent access to high bioavailability CR mimetic formats should
dispel such concerns, and besides, most of these food supplement molecules have
been used for centuries without such red flags arising in the medical literature.

Although CR/LE mimetics increase LE, they do so less than half as much as true

54
CR. There is a real difference between genuine CR and CR mimetics. In real CR,
the intracellular environment and extracellular environment are fuel depleted, while
under CR mimetic conditions, fuel and energy remain ad libitum in the extracellular
milieu. The CG pathway must shut down during real CR, but does it have to shut
down if only the CR/LE half of the system is strongly and directly affected? We
have already seen that TOR can be compartmentalized in the cell and behave
differentially [12]. All known CR mimetics operate on the CR/LE side. Maybe
down regulating CG with one of our silver medalists along with a CR mimetic
might force TOR to operate in all compartments like it must do in actual CR. In
addition, CG inhibition should assist activated CR in reducing carbohydrate
importation to the cell. Then, we might have a more realistic CR mimicry, and an
LE outcome more so approaching actual CR itself. Research of such a potential
synergistic effect on LE has yet to be conducted, but its effects on HE have been
well documented [25, 26, 29], and the use of CG inhibitors alone have had anti-
aging disease effects similar to CR mimetics, further indicating that such a synergy
should actually occur.

Our first primitive attempt to map the key elements of the CG/CM and CR/LE
system is just a beginning, and we hope that it is a correct beginning, because the
medical implications are altogether stupefying. If this really is that good beginning,
then it could function as a rational template for future pharmaceutical blockbusters,
concept pathway expansion to other diseases of aging that are known to be
favorably affected by CR mimetics and CG inhibitors, and a variety of, yet
unsuspected, or wholly unknown fruitful applications.

From our perspective, the system is coherent and consistent, and seems to explain a
lot, in terms if our present knowledge, especially in the way that so many pieces
seems to fit together, and so well. Only future research will tell if our ideas hold
together, as a global unifying hypothesis, for the furtherance in our understanding of
the aging diseases and life extension phenomena.

9) References

1. L. Brian Ray, Science, vol. 330, p. 1337, 12/3/2010.

2. Z. Feng, Cold Springs Harbor Laboratory Press, p. 199, 2010.

55
3. Seyfried and Shelton, Nutrition and Metabolism, vol. 7, no. 7, 1/27/2010.
4. G. Taubes, Science, vol.335, p. 28, 1/6/2012.
5. T. Ni Chonghaile, Science, vol. 334, p. 1129, 11/25/2011.
6. D. R. Green et. al., Science, vol. 333, p. 1109, 8/26/2011.
7. J. Rabinowitz and E. White, Science, vol. 330, p. 1344, 3/12/2010.
8. A. J. Levine and A. M. Puzio-Kuter, Science, vol.330, p. 1340, 3/12/2010.
9. Z. Szijgyarto et. al., Science, vol. 334, p. 802, 11/11/2011.
10. Jun Hee Lee et. al., Science, vol. 327, p. 1223, 3/5/2010.
11. J. Bass and J. S. Takahashi, Science, vol. 330, p. 1349, 3/12/2010.
12. R. Zonca et. al., Science vol. 330, p. 678, 11/4/2011.
13. J. C. Frankel, Science, vol334, p. 1194, 12/2/11.
14. E. H. Jennings et. al., Oncogene, vol. 29, p. 1056, 8/19/2010.
15. C. Canto et. al., Nature, vol. 458, p. 1056, 4/23/2009.
16. N. B. Ruderman et. al., Am. Jour. Physiol., Vol. 298, no. 4 E-751, April 2010.
17. Sundarsen et.al. Sci. Sig, ra 46, July 2011.
18. J. Patel et. al., Nutrition and Metabolism, Vol. 8, no. 7, 3/3/2011.
19. Kun-Liang Guang and Yu Xiong, Trends in Bioch. Sci., 2010.
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FACEBOOK.COM/LEPATH. Simply go to this web site, find the paper you wish to
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* * *

Gregory S. Bambeck Ph.D., e-mail: gregorybambeck@yahoo.com

Michael Wolfson J.D., M.B.A., e-mail: mwolfson@stanfordalumni.com

Warren Weller H. B., e-mail: warren.weller@yahoo.com

Copyright by Gregory S. Bambeck, Michael Wolfson and Warren Weller,

July 12, 2012

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