The Mitochondrial Factor: Key to Health


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Review the Microscopic Anatomy, Bioenergetics and Epigenetic Roles of the Mitochondria
Understand the Influence Mitochondria Play on Health Maintenance and Disease Prevention
Discuss Nonpharmacologic Approaches to Support Ideal Mitochondrial Function such as Diet, Supplementation and Exercise.

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The Mitochondrial Factor: Key to Health

  1. 1. The Mitochondrial Factor: Key to Wellness Andrew S. Bonci, BA, DC Private Practice Mission, Kansas
  2. 2. A Special Thank You ● And All of You! Missouri State Chiropractic Association ● Dr. Robert Riley ● Dr. Ron Manfredi ● Dr. Russell Matthias ● Dr. Doran Nicholson
  3. 3. Get the Notes Obtenga las Notas Akiru la Notojn or
  4. 4. Disclosures I work for NO special interest or lobbying groups nor do I receive stipends, consulting fees, gratuities or honoraria from any pharmaceutical or nutritional companies. 4
  5. 5. Most Humble Dedication When I sing to make you dance I truly know why there is music in leaves, and why waves send their chorus of voices to the heart of the listening earth. Rabindranath Tagore 1913 Nobel Laureate; Literature
  6. 6. Most Humble Acknowledgment Warburg and co-workers showed in the 1920s that, under aerobic conditions, tumor tissues metabolize approximately tenfold more glucose to lactate in a given time than normal tissues, a phenomenon known as the Warburg effect. Otto Heinrich Warburg (1883-1970) Nat Rev Cancer. 2011 May;11(5):325-37. 6
  7. 7. Objectives ● ● ● Review the Microscopic Anatomy, Bioenergetics and Epigenetic Roles of the Mitochondria Understand the Influence Mitochondria Play on Health Maintenance and Disease Prevention Discuss Nonpharmacologic Approaches to Support Ideal Mitochondrial Function such as Diet, Supplementation and Exercise. 7
  8. 8. The key to growth is the introduction of higher dimensions of consciousness into our awareness. ~Lao Tzu~
  9. 9. Minding My Own Business 9
  10. 10. And He Says ... It is well documented that tumorigenicity can be suppressed when cytoplasm from enucleated normal cells is fused with nucleated tumor cells to form cybrids. Seyfried, Thomas (2012-05-17T16:00:00+00:00). Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 5097-5099). Wiley Publishing. Kindle Edition.
  11. 11. Cybrids contain a single nucleus and mixtures of cytoplasm from two different cells. Seyfried, Thomas (2012-05-17T16:00:00+00:00). Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 5097-5099). Wiley Publishing. Kindle Edition.
  12. 12. The Secret of Cybrids
  13. 13. Suppression of Malignant State Israel and Schaeffer In Vitro Cell Dev Biol. 1987;23:627–32. showed that suppression of the malignant state could reach 100% in cybrids containing normal cytoplasm and tumorigenic nuclei. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 51065107). Wiley Publishing. Kindle Edition.
  14. 14. Nuclei Ain't Nothin' Israel and Schaeffer In Vitro Cell Dev Biol. 1988;24:487–90. also showed that nuclear/cytoplasmic hybrids derived by fusion of cytoplasts from malignant cells (nucleus absent) with karyoplasts from normal cells (nucleus present) produced tumors in 97% of the animals injected. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 51085109). Wiley Publishing. Kindle Edition.
  15. 15. The Secret of Cybrids
  16. 16. Whiskey Tango Foxtrot? It is also well documented that nuclei from cancer cells can be reprogrammed to form normal tissues when transplanted into normal cytoplasm despite the continued presence of the tumor-associated genomic defects in the cells of the derived tissues. Seyfried, Thomas (2012-05-17T16:00:00+00:00). Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 5176-5178). Wiley Publishing. Kindle Edition.
  17. 17. The Secret of Cybrids
  18. 18. It's About the OxPhos Normal mitochondria ... suppress tumorigenesis because their OxPhos is sufficient for maintaining energy homeostasis. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 52015202). Wiley Publishing. Kindle Edition.
  19. 19. The Differentiated State Respiration is required for the emergence and maintenance of differentiation, while loss of respiration leads to glycolysis, dedifferentiation, and unbridled proliferation. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 52555258). Wiley Publishing. Kindle Edition.
  20. 20. Hello! Is anyone listening? Replacement of damaged mitochondria with normal mitochondria, which will produce sufficient energy through respiration, restores the differentiated state. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 52605261). Wiley Publishing. Kindle Edition.
  21. 21. One More Time … Hello? In rephrasing, normal mitochondrial function maintains the differentiated state thereby suppressing carcinogenesis, whereas dysfunctional mitochondria can enhance dedifferentiation thereby facilitating carcinogenesis. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 52705272). Wiley Publishing. Kindle Edition.
  22. 22. “Complexity is a strategy used by professional elites to maintain control.” Ian Roberts The Energy Glut: The Politics of Fatness in an Overheating World, 2010. p.90
  23. 23. OxPhos = Mature State Thus, establishment of the mitochondrial system and engagement of oxidative metabolism are prerequisites for the differentiation of stem cells into a functional [cardiac] phenotype. Nat Clin Pract Cardiovasc Med. 2007 February; 4(Suppl 1): S60–S67.
  24. 24. Gylcolysis = Immature State Energy metabolism shifts from glycolysis to OxPhos with differentiation, or from OxPhos to glycolysis with reprogramming to pluripotency. Cell Stem Cell. 2012 Nov 2;11(5):589-95.
  25. 25. The Warburg Effect The Warburg effect underlies the simultaneous increase in glycolytic rate with a reduced mitochondrial respiratory rate; a kind of mitochondrial dysfunction that is found in tumor metabolic reprogramming pathways. Antioxid Redox Signal. 2012 May 15;16(10):1150-80.
  26. 26. Crabtree Effect Cancer cells have also been shown to be different from normal cells by reversibly down-regulating their oxygen consumption in response to increases in glucose: the Crabtree effect. PLoS One. 2013; 8(2): e56884. Increasing concentrations of glucose accelerates glycolysis (the breakdown of glucose) which results in the production of appreciable amounts of ATP through substrate-level phosphorylation. This reduces the need of oxidative phosphorylation done by the TCA cycle via the electron transport chain and therefore decreases oxygen consumption.
  27. 27. Joshua Lederberg, PhD The impediment to real discourse in academic communities is the tendency to proclaim what we know more than what we seek. Joshua Lederberg, PhD Nobel Prize Laureate Physiology and Medicine
  28. 28. The Gray Scale of Disease If it is true that a completely dysfunctional mitochondrial set results in cancer, then is it possible that a partially or incompletely dysfunctional mitochondrial set could result in any of the wide spectrum of diseases we see associated with genome instability … or “bad” genes?
  29. 29. Getting Reacquainted The word mitochondrion comes from the Greek μίτος, mitos, i.e. "thread", and χονδρίον, chondrion, i.e. "granule".
  30. 30. Endosymbiotic Theory The mitochondria are the product of a symbiosis between two micro-organisms that occurred about two billion years ago. The nature of the original partner organisms is actively debated, but the progenitor of the mitochondrion is thought to have been an alphaproteobacterium that harboured a complete OxPhos system. Philos Trans R Soc Lond B Biol Sci. 2013 July 19; 368(1622): 20120267.
  31. 31. Of Mitochondria and Chloroplasts The endosymbiotic theory of mitochondrial origin was substantiated with microbiological evidence by Lynn Margulis in 1967. J Theor Biol. 1967 Mar;14(3):255-74. Andreas Schimper (1883) that chloroplasts closely resembled free-living cyanobacteria.
  32. 32. Benefits of Symbiosis A single bacterial cell can generate only enough energy to sustain about 10,000 genes. When the host cell acquired multiple oxidative bacteria, the bacterial energy could be pooled to provide the required energy for adding more genes to the host cell’s DNA to create more complex anatomical structures. Philos Trans R Soc Lond B Biol Sci. 2013 July 19; 368(1622): 20120267.
  33. 33. Genome Expansion By enabling oxidative phosphorylation across a wide area of internal membranes, mitochondrial genes enabled a roughly 200,000-fold rise in genome size compared with bacteria. Nature. 2010 Oct 21;467(7318):929-34.
  34. 34. Costs of Gene Expression Whereas the energetic cost of possessing genes is trivial (2%), the cost of expressing them as protein (75%) is not and consumes most of the cell’s energy budget. Mitochondria increased the number of proteins that a cell can evolve, inherit and express by four to six orders of magnitude, but this requires mitochondrial DNA. Nature. 2010 Oct 21;467(7318):929-34.
  35. 35. The Price of Complexity Growth and replication of the nucleus is limited by mitochondrial energy production and thus calorie availability. Mitochondrion. 10(1): 12-31. Without mitochondria, [ cells ] cannot pay the energetic price of complexity. Nature. 2010 Oct 21;467(7318):929-34.
  36. 36. Mitochondria are of Female Origin The female oocyte (egg cell) will pass on more than 100,000 mitochondria to the next generation; sperm cells typically possess less than 100 mitochondria. Evans, Joseph (2013-02-28). The Secret Life of Mitochondria (Kindle Locations 136-138). Smashwords, Inc.. Kindle Edition.
  37. 37. What’s the Significance for Us? “Well, it means to find what is the source of your own life, and what is the relationship of your body, your physical form, to this energy that animates it. The body without the energy isn’t alive, is it? So you distinguish in your own life that which is of the body and that which is of energy and consciousness.” Campbell, Joseph; Bill Moyers (2011-05-18). The Power of Myth (p. 211). Knopf Doubleday Publishing Group. Kindle Edition.
  38. 38. The Giver of Forms “It’s the female as the giver of forms. She is the one who gave life to the forms and she knows where they came from. It is from that which is beyond male and female. It is from that which is beyond being and nonbeing. It both is and is not. It neither is nor is not. It is beyond all categories of thought and the mind.” Campbell, Joseph; Bill Moyers (2011-05-18). The Power of Myth (p. 226). Knopf Doubleday Publishing Group. Kindle Edition.
  39. 39. A Cellular Yin and Yang Mitochondria is the female energy and giver of forms. Nucleus is the male energy or action and creativity.
  40. 40. A Cellular Adam and Eve Eve … giver of forms … Adam … consciousness ...
  41. 41. A Cellular Shiva and Shakti Shakti … mitochondrial … giver of forms Shiva … nuclear … creative consciousness
  42. 42. Power and Wisdom is a Female Principle in Religion, Myth and Culture.
  43. 43. "I awoke, only to see the rest of the world was still asleep." Leonardo Davinci
  44. 44. 15 Pounds of Mitochondria Adults possess approximately 10 million billion mitochondria, which corresponds to approximately 10% of our body weight! Evans, Joseph (2013-02-28). The Secret Life of Mitochondria (Kindle Locations 138-139). Smashwords, Inc.. Kindle Edition.
  45. 45. Mitochondrial Bioenergetics
  46. 46. Mitochondrial Microanatomy There are five distinct parts to a mitochondrion. 1. outer mitochondrial membrane 2. intermembrane space (the space between the outer and inner membranes) 3. the inner mitochondrial membrane 4. the cristae space (formed by infoldings of the inner membrane) 5. the matrix (space within the inner membrane) Alberts, Bruce; Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, Peter Walter (1994). Molecular Biology of the Cell. New York: Garland Publishing Inc. ISBN 0-8153-3218-1.
  47. 47. Mitochondrial Outer Membrane The outer mitochondrial membrane contains enzyme systems for glycolysis, cardiolipin and phospholipid synthesis. Carolyn D. Berdanier. Mitochondria in Health and Disease CRC Press, 2005.
  48. 48. Glycolysis
  49. 49. Glycolysis Trivia Glycolysis produces its energy 100 times faster than aerobic respiration. Glycolysis is the preferred energy source of rapidly growing cell populations. The early zygote becomes increasingly dependent on glycolytic energy production as development progresses to the blastocyst stage. Int. J. Dev. Biol. 56: 799-808 (2012)
  50. 50. Mitochondrial Matrix The mitochondrial matrix contains enzyme systems for the Krebs cycle and ketone metabolism. The matrix also anchors the mitochondrial DNA (mtDNA). Carolyn D. Berdanier. Mitochondria in Health and Disease CRC Press, 2005.
  51. 51. Citric Acid Cycle
  52. 52. Mitochondrial Inner Membrane The inner mitochondrial membrane contains enzyme systems for partial fatty acid oxidation and the electron transport chain. Carolyn D. Berdanier. Mitochondria in Health and Disease CRC Press, 2005.
  53. 53.
  54. 54. NAD+/NADH The mitochondrial NADH is oxidized by the electron transport chain, which pumps protons across a membrane and generates ATP through oxidative phosphorylation. Biochem Soc Trans. 2003 Dec;31(Pt 6):1095105.
  55. 55.
  56. 56. FAD/FADH2 The primary biochemical role of FADH2 in eukaryotes is to carry high-energy electrons used for oxidative phosphorylation.
  57. 57.
  58. 58. Coenzyme Q10 CoQ10 functions as an electron carrier from enzyme complex I and enzyme complex II to complex III in this process … a process which also involves menaquinones or Vitamin K2. Science 336, 1306 (2012)
  59. 59.
  60. 60. Cytochrome C Cytochrome c carries an electron in the ETC and is also an intermediate in apoptosis, a controlled form of cell death used to kill cells in the process of development or in response to infection or DNA damage. Cell. 1996 Jul 12;86(1):147-57.
  61. 61. Cardiolipin: Mitochondrial-Specific Lipid Cardiolipin is a complex, mitochondrial-specific phospholipid that regulates numerous enzyme activities, especially those related to OxPhos and coupled respiration. Seyfried, Thomas (2012-05-17T16:00:00+00:00). Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 2125-2129). Wiley Publishing. Kindle Edition.
  62. 62. Cardiolipin: ETC Caulking Cardiolipin (CL) which represents 20% of membrane phospholipids is necessary for maintaining coupled mitochondria, and defects in CL can produce protein independent uncoupling. Hence, alterations in the content or composition of CL will alter cellular respiration. Seyfried, Thomas (2012-05-17T16:00:00+00:00). Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 2125-2129). Wiley Publishing. Kindle Edition.
  63. 63.
  64. 64. Mitochondrial Distribution Mitochondria are distributed throughout the cytoplasm in a nonrandom manner by cytoskeleton motors. Cells that are rapidly dividing have mitochondria close to the nucleus and to ribosomes because de novo protein synthesis is highly dependent on the energy provided by ATP. Carolyn D. Berdanier. Mitochondria in Health and Disease CRC Press, 2005.
  65. 65. Mitochondrial Genome The mitochondrial genome contains 37 genes that encode 13 proteins, 22 transfer RNAs, and 2 ribosomal RNAs. The 13 mitochondrial gene-encoded proteins all instruct cells to produce protein subunits of the enzyme complexes of the oxidative phosphorylation system, which enables mitochondria to act as the powerhouses of our cells. Nature Education 1(1):217; 2008.
  66. 66. mtDNA Mutations Most pathogenic mitochondrial DNA (mtDNA) mutations induce defects in mitochondrial oxidative phosphorylation (OxPhos). Epigenetics. 2012 Apr;7(4):326-34.
  67. 67. ROS mtDNA Dysfunction The mitochondrial free radical theory postulates that aging is caused by the toxicity of ROS, initiating a vicious cycle whereby damage to mtDNA and other mitochondrial constituents leads to respiratory chain dysfunction, which in turn leads to increased generation of ROS further facilitating mtDNA damage and thus creating a self-amplifying deterioration. J Intern Med. 2013 June; 273(6): 529–543.
  68. 68. Reactive Oxygen Species In general, harmful effects of reactive oxygen species on the cell are most often: ● ● ● ● damage of nDNA and mtDNA oxidations of polyunsaturated fatty acids in lipids (lipid peroxidation) oxidations of amino acids in proteins oxidatively inactivate specific enzymes by oxidation of co-factors
  69. 69. Antioxidant Defenses Antioxidant defenses are also up-regulated in conjunction with OxPhos to offset the effects of increased mitochondrial ROS production. Mitochondrion. 10(1): 12-31. Endogenous, genetically encoded antioxidant systems linked to the Antioxidant Response Elements (AREs) will be discussed in future slides. Exogenous antioxidants will be given passing attention in favor of the more powerful and evolutionary significant endogenous antioxidants.
  70. 70. Drug-induced Mitochondrial Toxicity Drugs that injure mitochondria usually do so by inhibiting respiratory complexes of the electron chain; inhibiting or uncoupling oxidative phosphorylation; inducing mitochondrial oxidative stress; or inhibiting DNA replication, transcription or translation. It is important to test for mitochondrial toxicity early in drug development as impairment of mitochondrial function can induce various pathological conditions that are life threatening or can increase the progression of existing mitochondrial diseases. Expert Opin Drug Metab Toxicol. 2005 Dec;1(4):655-69.
  71. 71. NSAID Induced Mitochondrial Toxicity [Mitochondrial toxicity] involves specific biochemical damage of mitochondria and uncoupling the oxidative phosphorylation reaction. Electron microscopic studies show vacuolisation and ballooning of mitochondria within an hour of indomethacin administration which is highly characteristic of uncoupling of oxidative phosphorylation. The consequence of uncoupling is diminished cellular ATP, which alters the intercellular junction, increases intestinal permeability, and releases calcium into cytosol which in turn causes secondary biochemical damage. Gut. 2001 February; 48(2): 163–167.
  72. 72. Mitochondrial Dysfunction Injury to the mitochondrial electron transport chain (ETC) or mutations of mtDNA leading to mitochondrial dysfunction have recently been suggested as an important factor in the pathogenesis of mitochondrial … a range of other diseases, aging, cancer and a range of other human disorders. human disorders. Epigenetics. 2012 Apr;7(4):326-34. Biochim Biophys Acta. 2010
  73. 73. Muscles Common Symptoms: Hypotonia, weakness, cramping, muscle pain, ptosis, and ophthalmoplegia. Despite the subjective weakness, many patients have minimal objective findings, possibly because fatigability is difficult to quantify in a physician’s office. Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  74. 74. Fibromyalgia? The etiology is unknown; however, recent studies suggest that mitochondrial dysfunction is involved in the pathophysiology of fibromyalgia. Mitochondrial DNA content (mtDNA/gDNA ratio) was reduced in fibromyalgia patients versus healthy controls (p<0.001). Expression levels of peroxisome proliferator-activated receptor gamma-coactivator 1-alpha was significantly lower in fibromyalgia patients (p<0.001) compared with healthy controls. Antioxid Redox Signal. 2013 Nov 20;19(15):1855-60.
  75. 75. Brain Common Symptoms: Developmental delay, mental retardation, autism, dementia, seizures, neuropsychiatric disturbances, atypical cerebral palsy, atypical migraines, stroke and stroke-like events. Migraine, dementia, seizures, and strokelike episodes can occur at any stage of the disease, but like myopathy, brain involvement is not required for the diagnosis. Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  76. 76. Nerves Common Symptoms: Neuropathic pain and weakness (which may be intermittent), acute and chronic inflammatory demyelinating polyneuropathy, absent deep tendon reflexes, neuropathic gastrointestinal problems (gastroesophageal reflux, constipation, bowel pseudoobstruction), fainting, absent or excessive sweating, and aberrant temperature regulation. Nerve cells and Schwann cells are extremely active metabolically: nerve cells require a tremendous amount of energy to maintain the electrochemical gradient necessary for nerve transmission. Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  77. 77. Kidneys Common Symptoms: Proximal renal tubular dysfunction, may result in loss of protein (amino acids), magnesium, phosphorous, calcium, and other electrolytes. The proximal renal tubular cells require an abundant and steady energy supply. Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  78. 78. Heart Common Symptoms: Cardiac conduction defects (heart blocks) and cardiomyopathy. The sinoatrial and atrioventricular nodes are the most metabolically active tissues in the body, and the muscular activity of the heart never ceases. Therefore, cardiac conduction defects and cardiomyopathy are complications of mitochondrial dysfunction. Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  79. 79. Liver Common Symptoms: Hypoglycemia, gluconeogenic defects, and nonalcoholic liver failure. Maintenance of glucose homeostasis is the most vital moment-to-moment function of the liver. Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  80. 80. Eye and Ear Common Symptoms: Optic neuropathy and Sensoryneural hearing loss Starting with high-frequency hearing loss, it can progress to total deafness. A number of mtDNA point mutations are associated with an extreme otosensitivity to aminoglycoside antibiotics (streptomycin). Cleve Clin J Med. 2001 Jul;68(7):625-6, 629-42.
  81. 81. Hope Springs Eternal A wide range of seemingly unrelated disorders have underlying pathophysiological mechanisms in common, namely reactive oxygen species (ROS) production, the accumulation of mitochondrial DNA (mtDNA) damage, resulting in mitochondrial dysfunction. If in the next 50 years advances in mitochondrial treatments match the immense increase in knowledge about mitochondrial function that has occurred in the last 50 years, mitochondrial diseases and dysfunction will largely be a medical triumph. Exp Mol Pathol. 2007 Aug;83(1):84-92.
  82. 82. Handle the difficult while it is still easy. Handle the big while it is still small. Difficult tasks begin with what is easy. Great accomplishments begin with what is small. Therefore, the wise never strive for the great and thus achieve greatness. Lao Tzu Tao Te Ching Chapter 63
  83. 83. Genome Instability If damage to mitochondria is persistent and defective mitochondria accumulate in the cell, it would lead to instability of the nuclear genome. T Hum Genet. 2009;54:647–54. Gene. 2005;354:140–6 Nucleic Acids Res. 2003;31:3909–17 Mutat Res. 2007;625:112–24 Epigenetics. 2012 Apr;7(4):326-34
  84. 84. Genome Instability Accumulated nuclear genome instability may help cells acquire new functions such as resistance to apoptosis, migration, and invasive characteristics which, in turn, could induce tumorigenesis. Epigenetics. 2012 Apr;7(4):326-34
  85. 85. Epigenetic Influences Evidence that mitochondrial dysfunction is associated with epigenomic changes can be found for a wide range of classical epigenomic diseases. In cancer, a genetic disease, changes in the epigenome such as loss of Imprinting (phenomenon by which certain genes can be expressed in a parent-of-origin-specific manner is lost) and hypomethylation are common. Mitochondrion. 10(1): 12-31.
  86. 86. Mitochondria & nDNA Methylation An increasing number of studies have identified a possible effect on the epigenetic landscape of the nuclear genome as a consequence of mitochondrial dysfunction. In particular, these studies demonstrate reversible or irreversible changes in genomic DNA methylation profiles of the nuclear genome. Epigenetics. 2012 Apr;7(4):326-34.
  87. 87. TCA Substrate & Epigenetics Co-factors, including FAD (Flavin Adenine Dinucleotide) Cell. 2004;119:941–53, acetyl–CoA Mitochondrion. 2010;10:12–31, Science. 2009;324:1076–80 and αketoglutarate (α-KG) are associated with the processes of active de/methylation or de/acetylation. Epigenetics. 2012 Apr;7(4):326-34 Reduced levels of FAD and α-KG due to mitochondrial impairment/dysfunction could have significant effects on regulation of the nuclear genome. Eur J Biochem. 2000;267:4888–900 Epigenetics. 2012 Apr;7(4):326-34.
  88. 88. Epigenetic Influences The chromatin modifications of cancer cells can be directly modulated by mitochondrial OxPhos. Cancer Biology and Therapy. 2008;7:1191–1193 Cancer Biology and Therapy. 2008;7:1182–1190.
  89. 89. Hello? Was Anybody Listening? Normal mitochondria ... suppress tumorigenesis because their OxPhos is sufficient for maintaining energy homeostasis. Seyfried, Thomas. Cancer as a Metabolic Disease: On the Origin, Management, and Prevention of Cancer (Kindle Locations 52015202). Wiley Publishing. Kindle Edition.
  90. 90. Then said a teacher, "Speak to us of Teaching." And he said: No man can reveal to you aught but that which already lies half asleep in the dawning of our knowledge. ~Khalil Gibran: The Prophet~
  91. 91. MtDNA Rehabilitation When mtDNAs were reintroduced into the mtDNAdeficient cancer cells, 30% of the hypomethylated sites become remethylated. Thus, there is a direct cause and effect relationship between mitochondrial function and epigenomic methylation in cancer cells. Cancer Biology and Therapy. 2008;7:1191–1193 Cancer Biology and Therapy. 2008;7:1182–1190.
  92. 92. OxPhos-p53 Connection The p53 protein also regulates OxPhos and it has been shown that mutational inactivation of p53 in cancer cells suppresses OxPhos and induces glycolysis. Science. 2006;312:1650–1653.
  93. 93. If mitochondria are so critical for cellular and overall health of the organism, then it behooves us to focus ourselves on mitochondrial biogenesis as a way to foster overall health of the organism.
  94. 94. Mitochondrial Biogenesis Mitochondria cannot be generated de novo; instead, they proliferate by growth and division of pre-existing organelles. Nature Rev Mol Cell Biol. 2010;11:872–884. Mitochondria divide during mitosis, providing daughter cells with a normal complement of mitochondria. Antioxid Redox Signal. 2012 May 15;16(10):1150-80 There are, however, instances in which mitochondrial divisions are not tied to cell cycle. Antioxid Redox Signal. 2012 May 15;16(10):1150-80
  95. 95. Mitochondrial Biogenesis Nutrient excess stimulates mitochondrial fission, fragmented mitochondrial morphology, and an arrest in oxygen consumption, oxidative phosphorylation, and Cr Cra abt ATP synthesis. Front Aging Neurosci. 2013 Sep 6;5:48. btre ree eE By contrast, nutrient deficiency triggers mitochondrial Effffe ect ct fusion, elongated mitochondrial morphology, and an acceleration of mitochondrial respiration and ATP production. Front Aging Neurosci. 2013 Sep 6;5:48. Thus, mitochondrial fission and fusion is a fine tuned process that controls the switches of energy production with energy demand, thereby maintaining homeostasis. Commun Integr Biol. 2011 November 1; 4(6): 752–754.
  96. 96. The Biogenesis Big Three 1.Calorie Restriction and Fasting 2.Dietary Polyphenols 3.Endurance Exercise
  97. 97. Calorie Restriction/ Fasting SIRT1 Dietary Polyphenols Endurance Exercise
  98. 98. SIRT1 SIRT1 stands for silent information regulator type 1. SIRT1 is an enzyme that deacetylates proteins that contribute to cellular regulation (reaction to stressors, longevity). J Biol Chem. 2005 Apr 22;280(16):16456-60.
  99. 99. CR/Fasting Targets SIRT1 Sirt1 is activated by calorie restriction in mammals and acts as a trigger in mitochondrial biogenesis. Am J Physiol Regul Integr Comp Physiol. 2011 Jul;301(1):R67-75. BONUS: Upon food withdrawal Sirt1 protein binds to and represses genes controlled by the fat regulator PPAR-γ (peroxisome proliferator-activated receptor-γ), including genes mediating fat storage. Nature. 2004 June 17; 429(6993): 771.
  100. 100. Polyphenols Target SIRT1 Naturally occurring dietary polyphenols, such as resveratrol, curcumin, quercetin, and catechins, have antioxidant and anti-inflammatory properties via modulating different pathways. In addition, these polyphenols have also been shown to activate SIRT1 directly or indirectly in a variety of models. Arch Biochem Biophys. 2010 September 1; 501(1): 79–90.
  101. 101. Exercise Targets SIRT1 SIRT1 is increased in response to aerobic exercise. Metabolism 2008; 57: 986-998. Rejuvenation Res 2008; 11: 139-150. Unlike the increase in mitochondrial content observed with regular aerobic exercise, resistance training, such as lifting weights, is associated with an increase in the contractile machinery. Exercise Physiology:Energy, Nutrition, and Human Performance. 5th ed. Baltimore: Lippincott Williams & Wilkins; 2001 p. 500-547. This increase in the production of contractile proteins results in increased muscle mass with very little enhancement of mitochondrial production.
  102. 102. Calorie Restriction/ Fasting SIRT1 Dietary Polyphenols PGC-1α Endurance Exercise
  103. 103. PGC-1α: Master Regulator PGC-1α = Peroxisome proliferator-activated receptor gamma coactivator 1-alpha PGC-1α appears to act as a master regulator of mitochondrial biogenesis by integrating and coordinating the activity of multiple transcription factors. Cell 1998;92(6):829–39. Expression levels of PGC-1α are directly related to mitochondrial biogenesis activity. J Cell Sci. 2012 Nov 1;125(Pt 21):4963-71
  104. 104. Calorie Restriction/ Fasting SIRT1 Dietary Polyphenols PGC-1α Endurance Exercise Mitochondrial Biogenesis
  105. 105. Benefits of Mitochondrial Biogenesis Increased Decreased ● OxPhos ● ROS ● Metabolic Efficiency ● Oxidative Stress ● Energy Level ● Body Fat ● Lean Body Mass ● Disease/Dysfunction ● Exercise Performance ● Age-related Deterioration ● Life Span (?)
  106. 106. Calorie Restriction In 1935, nutritional scientist Professor Clive M. McCay (Cornell University) published his seminal paper showing that rats fed a diet with 30-40% fewer calories lived about 33% longer than rats fed ad libitum. Proc Nutr Soc 1995; 54: 657-664.
  107. 107. The Responsible Component? From the beginning, investigators raised the question of whether it was caloric restriction or a specific dietary component such as fat, protein, carbohydrate, vitamins, or antioxidants that was responsible for observed physiological and metabolic benefits. After manipulating the various dietary components it was found that health and longevity effects derived predominantly from calorie restriction alone. Proc Nutr Soc 1995; 54: 657-664.
  108. 108. CRON-diet Calorie Restriction Optimal Nutrition The CRON-diet involves calorie restriction while still attempting to provide the recommended daily amounts of various nutrients. Proponents recommend a goal of restricting intake by 20%. The actual daily amount eaten depends on the adherent's basal metabolic rate (BMR). A common daily intake is 1800 calories per day.
  109. 109. Roy Lee Walford, M. D. (1924-2004) Walford found that mice fed a CRON of 50% normal caloric intake almost doubled their expected life span. Walford was one of eight “crew members” sealed inside Biosphere 2 where they lived from Sept 26, 1991 to Sept 26, 1993.
  110. 110. 20 Years of CR/CRON Research [P]rolonged CR appears to extend the lifespan of rhesus monkeys, which exhibited lower body fat; slower rate of muscle loss with age; lower incidence of neoplasia, cardiovascular disease, type 2 diabetes mellitus, and endometriosis; improved insulin sensitivity and glucose tolerance; and no apparent adverse effect on bone health, as well as a reduction in total energy expenditure. In addition, there are no reports of deleterious effects of CR on reproductive endpoints, and brain morphology is preserved by CR. ILAR J. 2011 February 8; 52(1): 66–77.
  111. 111. CRON Weakness Calorie restrictors are exquisitely aware of exactly how many calories they are eating at all times. CRON, in other words, falls prey to one of the biggest stumbling blocks for any diet: It requires you to constantly think about the one thing you don’t want to be thinking about: what you can and can’t eat. Johnson M.D., James B. (2008-04-10). The Alternate-Day Diet: Turn on Your "Skinny Gene," Shed the Pounds, and Live a Longer and HealthierLife (Kindle Locations 242-244). Penguin Group US. Kindle Edition.
  112. 112. Intermittent Fasting Intermittent fasting (IF) is a pattern of eating that alternates between periods of fasting (usually meaning consumption of water and sometimes low-calorie drinks such as black coffee) and non-fasting.
  113. 113. Intermittent Fasting Interestingly, intermittent fasting but not caloric restriction for 20 weeks increases hippocampal neuron tolerance to excitotoxic stress in mice, suggesting neuroprotective effects of intermittent fasting. Proc Natl Acad Sci U S A 100: 6216–6220.
  114. 114. Alternate Day Fasting Calorie restriction (CR) and alternate-day fasting (ADF) represent 2 different forms of dietary restriction (DR). The findings in animals suggest that ADF may effectively modulate several risk factors, thereby preventing chronic disease, and that ADF may modulate disease risk to an extent similar to that of CR. Am J Clin Nutr. 2007 Jul;86(1):7-13.
  115. 115. Eat Frequent, Small Meals (?) The idea that eating little and often is a “good thing” has been driven partly by snack manufacturers and faddish diet books, but it has also had support from the medical establishment. Their argument is that it is better to eat lots of small meals because that way we are less likely to get hungry and gorge on high-fat junk. Mosley, Michael; Spencer, Mimi (2013-02-26). The FastDiet: Lose Weight, Stay Healthy, and Live Longer with the Simple Secret of Intermittent Fasting (p. 14). Atria Books. Kindle Edition.
  116. 116. Eating Between Meals Once upon a time, parents told their children, “Don’t eat between meals.” Recent research in the United States, which compared the eating habits of 28,000 children and 36,000 adults over the last thirty years, found that the amount of time between what the researchers coyly described as “eating occasions” has fallen by an average of an hour. In other words, over the last few decades the amount of time we spend “not eating” has dropped dramatically. Mosley, Michael; Spencer, Mimi (2013-02-26). The FastDiet: Lose Weight, Stay Healthy, and Live Longer with the Simple Secret of Intermittent Fasting (p. 14). Atria Books. Kindle Edition.
  117. 117. Fasting is Evolutionary For most animals out in the wild, periods of feast or famine are the norm. Our remote ancestors did not often eat four or five times a day. Instead they would kill, gorge, lie around, and then have to go for long periods of time without having anything to eat. Our bodies and our genes were forged in an environment of scarcity, punctuated by the occasional massive blowout. Mosley, Michael; Spencer, Mimi (2013-02-26). The FastDiet: Lose Weight, Stay Healthy, and Live Longer with the Simple Secret of Intermittent Fasting (p. 13). Atria Books. Kindle Edition.
  118. 118. Seasonal Expressions of Mitochondria When carbohydrates were abundant to our ancestors, such as during the plant growing season, consumption of plant starch resulted in increased blood glucose levels. When glucose is abundant PGC-1α is down-regulated, mitochondrial OxPhos is depressed, and metabolism shifts toward glycolysis. Mitochondrion. 10(1): 12-31.
  119. 119. Insulin-Metabolism Response A rise in serum glucose stimulates the pancreatic β-cells to release insulin. Insulin signals the availability of carbohydrates to the rest of the tissues in the body, shifting metabolism toward glycolysis for ATP generation Mitochondrion. 10(1): 12-31., the storage of excess calories as fat by activating lipoprotein lipase J Clin Invest. 1982 May; 69(5): 1119–1125. and conservation of adipose tissue by the suppression of cellular [hormone-sensitive] lipase Mol Cell Biol. 2010 November; 30(21): 5009–5020..
  120. 120. Mitochondrial Response to Glucagon When carbohydrates are scarce, glucose becomes limiting to animals, serum glucose levels fall, insulin secretion declines, and the pancreatic α-cells secrete glucagon. Glucagon then signals via cAMP to mobilize stored fat and up-regulate mitochondrial OxPhos to burn fat and generate energy to survive periods of glucose deprivation. Annual Review of Biochemistry. 2007;76:781–821.
  121. 121. Carbohydrate Scarcity & PGC-1α [W]hen carbohydrates are limited, the expression of PGC-1α is doubly induced switching cellular metabolism toward OxPhos to burn stored fats to survive periods of carbohydrate deprivation. Mitochondrion. 10(1): 12-31.
  122. 122. Beta-Hydroxybutyric Acid Beta-hydroxybutyrate, the principal "ketone" body in starving man, displaces glucose as the predominating fuel for the brain, decreasing the need for glucose synthesis in the liver (and kidney) and accordingly spares its precursor, muscle-derived amino acids. Trans Am Clin Climatol Assoc. 2003; 114: 149–163.
  123. 123. Keystone Ketone of Evolution [E]volution occurs in long stable periods (equilibrium) interrupted by brief and rapid changes (punctuation). This could not have happened without betahydroxybutyric acid supporting survival as a fuel source during the frequent famines caused by natural catastrophies as well as those caused by man himself during hostilities and migrations. Trans Am Clin Climatol Assoc. 2003; 114: 149–163.
  124. 124. Metabolic Efficiency Decreasing food intake decreases the UCPs, “coupling” is tighter, and metabolic efficiency is increased. Increasing the UCPs increases the metabolic rate which tends to reduce obesity, but it generates more free radicals. Walford R. Beyond the 120 Year Diet, 2000. ISBN-10: 1568581572. p. 63.
  125. 125. Fasting State is Protective Ketones inhibit mitochondrial production of ROS by enhancing NADH oxidation in the mitochondrial respiratory chain. Neuroscience. 2007 March 2; 145(1): 256–264. Studies in cardiac tissue have suggested that ketones reduce oxidative stress IUBMB Life. 2001;51:241–247, a pathogenic process implicated in many disorders ranging from atherosclerosis and traumatic injuries to diseases more specific to the nervous system Physiol Rev. 2002;82:47–95; Neurology. 2005;64:1152–1156.
  126. 126. Ketones: the Superfuel Recent studies have shown that beta-hydroxybutyrate, the principal "ketone", is not just a fuel, but a "superfuel" more efficiently producing ATP energy than glucose or fatty acid. In a perfused rat heart preparation, it increased contractility and decreased oxygen consumption. Trans Am Clin Climatol Assoc. 2003; 114: 149–163.
  127. 127. Successful Fasting as a Selective Pressure A normal 70 kg man survives 2-3 months of starvation [on ketones] instead of several weeks, and obese man many months to over a year. Without this metabolic adaptation, homo sapiens could not have evolved such a large brain. Trans Am Clin Climatol Assoc. 2003; 114: 149–163.
  128. 128. Metabolic Efficiency Take in fewer calories and your body increases its metabolic efficiency; up your intake and your efficiency decreases. Walford R. Beyond the 120 Year Diet, 2000. ISBN-10: 1568581572. p. 63.
  129. 129. Ketoacidosis In the super-fasted state (the diabetic) where even basal insulin levels are inadequate, excessive amino acids are released from muscle, hepatic gluconeogenesis and ketogenesis increase and levels of glucose and ketones rise. These result in progressive hyperglycemia, hyperketosis and glucosuria and ketonuria. Diabetes Metab Res Rev 1999;15:412-26.
  130. 130. Ketosis v. Ketoacidosis Benign dietary ketosis is a controlled, insulin regulated process which results in a mild release of fatty acids and ketone body production in response to low carbohydrate intake. pH below 7.3 Ketoacidosis occurs when the body is producing large quantities of ketone bodies via the metabolism of fatty acids (ketosis) and the body is producing insufficient insulin to slow this production. pH below 7
  131. 131. Simple Starvation Simple starvation and/or the ketogenic diet produce a mild but closely regulated metabolic acidosis, compatible with normal life as evidenced. Trans Am Clin Climatol Assoc. 2003; 114: 149–163.
  132. 132. CR/Fasting/Ketosis Protective Effects Animal models and isolated cells show that ketone bodies, especially β-hydroxybutyrate, confer neuroprotection against diverse types of cellular injury. Behav Pharmacol. 2006 September; 17(5-6): 431–439. Nrf2-regulated signaling pathway is critical in protecting mitochondria from oxidative stress during feed deprivation, which ensures efficient utilization of fatty acids. PLoS One. 2013; 8(3): e59122.
  133. 133. Nrf2 Triggers During periods of oxidative stress, Nrf2 is released from sequestration in the cytoplasm and translocates to the nucleus. Nrf2 binds antioxidant response elements (AREs) in the regulatory regions of target genes and activates transcription. Toxicol Pathol. 2007 Jun;35(4):459-73. The Nrf2 antioxidant response pathway is "the primary cellular defense against the cytotoxic effects of oxidative stress." N Engl J Med. 2012 Sep 20;367(12):1098-107. Integr Comp Biol. 2010 November; 50(5): 829–843.
  134. 134. Three Important Nrf2 Targets 1. Superoxide Dismutase 2. Catalase 3. Glutathione Peroxidase
  135. 135. Superoxide Dismutase SODs are enzymes that catalyze the breakdown of the superoxide anion into oxygen and hydrogen peroxide. Free Radic Biol Med. 2002 Aug 1;33(3):337-49. CRC Crit Rev Biochem. 1987;22(2):111-80. SOD enzymes are present in almost all aerobic cells and in extracellular fluids. Mol Aspects Med. 2005 Aug-Oct;26(4-5):340-52.
  136. 136. Superoxide Dismutase Superoxide dismutase enzymes contain metal ion cofactors that can be copper, zinc, manganese or iron. In humans, the copper/zinc SOD is present in the cytosol, while manganese SOD is present in the mitochondrion. CRC Crit Rev Biochem. 1987;22(2):111-80. There also exists a third form of SOD in extracellular fluids, which contains copper and zinc in its active sites. Int J Biochem Cell Biol. 2005 Dec;37(12):2466-71.
  137. 137. Catalase Catalases are enzymes that catalyse the conversion of hydrogen peroxide to water and oxygen, using either an iron or manganese cofactor. Cell Mol Life Sci 61 (2): 192–208. Prog Biophys Mol Biol 72 (1): 19–66. This protein is localized to peroxisomes in most eukaryotic cells. Free Radic Biol Med 13 (5): 557–80.
  138. 138. Glutathione Peroxidase Glutathione peroxidase is an enzyme containing four selenium-cofactors that catalyzes the breakdown of hydrogen peroxide and organic hydroperoxides. Free Radic Biol Med 27 (9–10): 951–65.
  139. 139. Three Important Interventions Upregulate Nrf2 Expression 1.Dietary Restriction (DR) in the form of calorie restriction and fasting. 2.Dietary Polyphenols 3.Aerobic Exercise
  140. 140. CR/Fasting Targets Nrf2 Fasting induces NRF2 target gene expression by at least 1.5 to 5-fold. Antioxid Redox Signal. 2013 Jul 31. The Nrf2 antioxidant response pathway is "the primary cellular defense against the cytotoxic effects of oxidative stress." N Engl J Med. 2012 Sep 20;367(12):1098-107. Nrf2 is the primary transcription factor that binds to the antioxidant responsive elements (AREs) and activates the expression of these genes. It is evident that activation of ARE-regulated genes contributes to the regulation of cellular antioxidant defense systems. Curr Pharm Des. 2004;10(8):879-91.
  141. 141. Polyphenols Target Nrf2 Increasing Nrf2 expression or biological activity by small molecules such as polyphenols among others can be achieved in various cell types. Curcumin and resveratrol have been shown to activate Nrf2 leading to subsequent protection from oxidative insult and toxins. Flavonoid polyphenols such as epigallocatechin 3-gallate (EGCG) and quercetine are potent activators of Nrf2 that have been shown to be neuroprotective against oxidative stress in vitro. Recent Pat CNS Drug Discov. 2012 December; 7(3): 218–229.
  142. 142. Aerobic Exercise Targets Nrf2 Exercise activates Nrf2, the master regulator of antioxidant defenses, and attenuates CVD. Exerc Sport Sci Rev. 2013 Jul;41(3):162-8. Nrf2 is an important mediator of the antioxidant defense mechanism that transcriptionally regulates several phase II antioxidant enzyme-genes, including the glutathione complexes, superoxide dismutase and others. Am J Physiol Renal Physiol. 2009 November; 297(5): F1174–F1180. Integr Comp Biol. 2010 November; 50(5): 829–843.
  143. 143. Recommendations, or ...
  144. 144. Measure with a Micrometer
  145. 145. Mark with Chalk
  146. 146. Cut with an Axe
  147. 147. Hell! Cut with a Chainsaw!!
  148. 148. The Value of RCT Provides important information about an intervention's likely outcome in general practice. It should address margins of efficacy and safety. A sophisticated marketing strategy employed by professional elites to control the marketplace. Ian Roberts, PhD Statistical analysis is often used as a highly effective propaganda tool. David R. Hawkins, MD, PhD
  149. 149. Single Subject Research Design Single-subject research is a group of research methods that are used extensively in the experimental analysis of behavior and applied behavior analysis with both human and non-human participants. The most basic method in this type of research is the AB-A design.
  150. 150. Single Subject Research Design These methods form the heart of the data collection and analytic code of behavior analysis. Behavior analysis is data driven, inductive, and disinclined to hypothetico-deductive methods. Statistical methods have been largely ignored.
  151. 151. How Does One Begin?
  152. 152. Begin with a Conversation
  153. 153. What Does DR Look Like in Practice? It either looks like counting calories and restricting those calories over three meals each day, or It looks like intermittent fasting. I feel that intermittent fasting is preferable to traditional CR diets for several reasons. My major reason in particular is that it appears more accurately to reflect the large swath of our evolutionary existence and those adaptive mechanisms.
  154. 154. Carnivore? Herbivore? … or ... Omnivore built for scarcity This can explain the ● Dentition ● Digestive tract Length ● Gall Bladder Size
  155. 155. Which Supplements Do You Need to Take? Dark Berries Resveratrol Green Tea (ECGCs) Turmeric Vary them from time to time
  156. 156. What Really is Aerobic Exercise? Aerobic exercise is physical exercise of relatively low intensity that depends primarily on the aerobic energygenerating processes. Oxygen supply meets or exceeds the needs of exercising tissues.
  157. 157. Are there Synergistic Effects? Voluntary exercise and resveratrol (RSV) treatment induced mitochondrial biogenesis independent of each other. When RSV and exercise were combined, a synergistic effect was evident, leading to enhanced translocation of PGC-1α and SirT1 to the nucleus and stimulation of mitochondrial biogenesis. Thus, the magnitude of the effect of RSV on muscle mitochondrial biogenesis is reliant on the cellular environment and by repeated bouts of exercise. J Biol Chem. 2013 Mar 8;288(10):6968-79.
  158. 158. Thank You