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.

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

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
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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 costimulate 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 sugarphosphate 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
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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,
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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.
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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
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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
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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
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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
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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.

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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, E751, 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 postprandial 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 PGC1alpha, 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 coactivator 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 yinyang function. Just as TOR may have a CG-like function during CR, SIRT 1 may
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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 PGC1alpha 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
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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
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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.

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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
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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.
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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
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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 antioxidant, 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
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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
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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
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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
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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.

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Figure 2. Substrate and ATP flow alterations in the cancer cell, as presented in the 1981 Bambeck dissertation at Kent State University [27].
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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
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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,
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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 prediabetic 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
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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 fullblown 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.
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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
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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
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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
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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
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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
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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. 209406, 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.
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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
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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.

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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
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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.

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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
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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
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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
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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.

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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.
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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.
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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 multiten 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
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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 antiaging 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.
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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. 20. Hirschey Lab, lab.hirschey.org/research/files/acetylation, 2012. 21. J. Zang et. al., Molecular and Cellular Proteomics, vol. 8, p. 215 2/1/2009. 22. A. Coghlan, New Scientist, p. 6, 5/15/2010. 23. D. Anastasio, Science, vol. 334, p. 1278, 2/12/2010. 24. M. Jain et. al., Science, vol. 336, p. 1040, 5/25/2012. 25. 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. 26. lef.org/references. 27. G. Bambeck, Dissertation, Kent State University, 1981. 28. J. C. Cohen et. al., Science, vol. 332, p. 1519, 6/24/11. 29. Disease Prevention and Treatment, fourth edition, published by Life Extension Media, cancer: pp. 209-406, cardiovascular disease: pp. 209-406, diabetes II: pp. 709-742, obesity: pp. 1241-1272, 2003. 30. S. Bhaleral and T. R. Clandidin, Science, vol. 336, p. 1040, 6/8/2012.

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All of our SCRIBD publications can be easily two-step accessed via our web site at FACEBOOK.COM/LEPATH. Simply go to this web site, find the paper you wish to read, and click the link. The paper will automatically be accessed from SCRIBD, and appear on your screen. * * *

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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