A CURRENT UNDERSTANDING OF PARKINSON’S Although thousands of studies exist searching for answers as to the cause and cure of Parkinson’s, there seems to be a vast disconnect. It is as if no one has ever read enough of the studies to come to any valuable conclusions. I found in reviewing and comparing the scientific studies, however, that a connection does emerge between the studies, history, and what people are actually experiencing. In summary, it has become increasingly apparent that what is missing in the current understanding of Parkinson’s is this:1. Damage to the mitochondria,where in damaged mitochondria are reproducing damaged mitochondria is not just a “part” of Parkinson’s, but the basis. Not only is damaged mitochondria the basis for Parkinson’s, but also likely for many neurological diseases, diabetes, immunological diseases, and cancers.2. Mercury is the number one suspect today as that which is causing damage to mitochondria, leading to so many devastating diseases. This is because of mercury’s unfettered, widespread, even brazen use in dentistry, medicine, and industry. It’s a poison we are actually injecting into people where we would never dream of doing the same with other poisons in the same category (like lead or arsenic). But it is also the number one suspect, because mercury has been shown to have perhaps the highest affinity for doing damage exactly how and where the damage is being done in Parkinson’s.3. All the”things going on”that are being observed in Parkinson’s stem from the damaged mitochondria directly or indirectly. Continuing to focus upon, study, and address all the “things going on” without focusing upon and addressing the mitochondria is doomed to never cure Parkinson’s.4. Many diseases are thought or known to have mitochondrial damage as the basis. So when the day comes that man focuses upon prevention of mitochondrial damage while also doing that which heals the mitochondria we will likely see a revival of sorts in the medical/health fields, such as has not been seen in medicine since the discovery of “germs”, especially given the many diseases to which damaged mitochondria is now linked.ii | P a g e
TABLE OF CONTENTS MITOCHONDRIAL DAMAGE AND PARKINSON’S Different Types of Parkinson’s?
All That Is Going On In Parkinson’s Emanates From The Mitochondria Dopamine, One Of Many Neurotransmitters
Excitotoxicity: Glutamate & Nitric OxideGlutamateNitric Oxide
The Mitochondrial Connection MERCURY Forms of Mercury
But My Dentist Told Me Mercury Isn’t Toxic
How Mercury Damages The Mitochondria
Mercury And Complex I Of The Mitochondria
Reactive Oxygen Species From Damaged Mitochondria
Mercury And The Brain’s Immune System
Mercury And The Viral Connection
Some Other Toxins That Have Been Shown To Cause Parkinsonism Rotenone
The Acetogenin Family Of Compounds OrganophosphatesALL THE “THINGS GOING ON” IN PARKINSON’S TRACE BACK TO MITOCHONDRIAL DYSFUNCTION
Iron And Copper – Causal Or A Result?
Not Simply A Dopamine DeficiencyThe Parkinson’s Brain Is Toxic To Dopamine
Mitochondrial Membrane, Electron Transfer Chain Dysfunction Calcium Homeostasis
How Does Increased Cellular Calcium Lead To Toxicity? Genes, Genetic Mutations, Protein Mutationiii | P a g ePerturbed Endoplasmic Reticulum Function Immune System Disruption
Neurons Firing Wildly Out Of Control Glutathione DeficiencyUp-Regulation Of Gamma-Glutamyltranspeptidase Homocysteine
The Need For Ascorbic Acid
Are Drugs An Insult To Injury?CAN WE REPAIR DAMAGED MITOCHONDRIA?Mitochondrial Transplant
Stem Cell Therapy
Nutritional Supplements For Parkinson’s Hormetic Pathway – Vitagenes BioavailabilityAcetyl-L-Carnitine Alpha Lipoic Acid Alpha Tocopherol Ascorbic Acid AstaxanthinBlack Tea Extracts Caffeine
Carnosine Coenzyme Q10 Creatine
Curcumin Ganoderma Lucidum Ginkgo Biloba Ginsenoside L-TheanineNAD
Retinoids And Carotenoids Rhodiola Rosea
Silymariniv | P a g e
A Diet Rich In PhytochemicalsChart of Phytochemicals Specific Foods For HealingSweet Green Tea Muscadine Grapes Mangoes
GarlicSoybeansTomatoes Diagnostic TestsMercury Testing
Mitochondrial Dysfunction Tests
Urine Test Taken As First Morning After Fasting Red Flags
MitoSciencesClinical Mitochondrial Therapies
DMSA For Mercury Chelation
Methylcobalamin, Folate and B6 Mitochondria-Targeted Peptide AntioxidantsSS AntioxidantsRESOURCES INDEX
MITOCHONDRIAL DAMAGE AND PARKINSON’S It was in 175 AD that the physician Galen described the “shaking palsy”. It wasn’t until 1817 that a detailed medical essay was published on Parkinson’s by London doctor James Parkinson. We must ask ourselves, is there a single common denominator that connects Parkinson’s today to what was observed in centuries past?In 1991 an article in California’s Orange County Register exclaimed: “Cause of Parkinson’s disease may have been discovered.” Within the article they say “Scientists may have tracked down the long sought cause of Parkinson’s disease with the discovery of a defect in the ‘energy factories’ in muscles of people with the ailment.” 1 The “energy factory” is your mitochondria. Much study on the mitochondria has gone on since then. For one thing, we now know that Parkinson’s stems not from the muscles themselves, but from damage to mitochondria in the brain which leads to a cascade of continuous, even self- perpetuating events that damage both dopaminergic neurons and dopamine itself, which results in the various symptoms we see in Parkinson’s.1 Paul Raeburn “Cause of Parkinson’s disease may have been discovered” Simple methods may prevent illness, researchers say. Friday, August 2, 1991 The Orange County Register.1|Page
MITOCHONDRIAL ELECTRON TRANSPORT CHAIN
NADH dehydrogenase(or NADH: quinone reductase) is an enzyme located in the inner mitochondrial membrane that catalyzes the transfer of electrons from NADH to coenzyme Q. This is the entry enzyme of oxidative phosphorylation in the mitochondria
Succinate dehydrogenase (or succinate-coenzyme Q reductase) is an enzyme complex that exists in the inner mitochondrial membrane. It is the only enzyme that participates in both the citric acid cycle and the electron transport chain.
Coenzyme Q (cytochrome c-oxidoreductase, or cytochrome bc1 complex) is the third complex in the electron transport chain of the mitochondria. It’s role is that of generating ATP (adenosine triphosphate) through oxidative phosphorylation.
Cytochrome c oxidase is a large transmembrane protein complex found in the mitochondrial membrane. It receives an electron from each of four cytochrome c molecules, transferring them to one oxygen molecule, converting oxygen to two molecules of water. In the process it translocates four protons across the mitochondrial membrane to establish a transmembrane difference of proton “electrochemical potential” that ATP synthase then uses to make ATP.
Suffice it to say here that when the function of the mitochondria’s respiratory chain complexes are impaired, there is an enhanced production of superoxide anion3 creating “mitochondrial toxins” so often referred to in literature today. These toxins (all free radicals) include hydrogen peroxide (H2O2), peroxynitrite, and hydroxyl radical. All of these are created by, and turn around to do further damage to mitochondrial proteins and membrane permeability, leading to severe mitochondrial dysfunction and destruction as well as “all the things going on” in Parkinson’s and other neurological diseases.One mitochondrial toxin is excess nitric oxide. Some who are treating Parkinson’s are using therapies and supplements that increase nitric oxide when it is already being generated in excess. In fact, high levels of neuronal and inducible nitric oxide synthase were found in the substantia nigra of patients and animal models of Parkinson’s disease.4 S-Nitrosation of mitochondrial proteins appears to contribute to the negative interactions of nitric oxide and its derivatives with the mitochondria. When the mitochondria’s supply of glutathione (an antioxidant produced within the body) is minimal or gone, the mitochondria has little defense against nitric oxide. Thus we have mitochondria reproducing damaged mitochondria and production of oxygen free radicals interacting with nitric oxide and the resultant production of peroxynitrite and a vicious cycle is perpetuated. So what is it that can do such damage to the mitochondria?The one damaging toxin around when Parkinson’s was observed in 175 AD and still around during James Parkinson’s observations in the 1800s and still being used today, is the toxic metal mercury. We will discuss more later exactly why mercury is so toxic; how it does its damage; how it is the initial toxic agent; exactly what damage it does; as well as all the subsequent damage done by the “fallout” of mercury damage.Of course there are other toxic substances, especially used in labs, that can cause parkinsonian symptoms if a person is injected with or poisoned by them. For now we’re labeling mercury as our “number one” suspect. And this might be an3 Turrens JF. Superoxide production by the mitochondrial respiratory chain. Biosci Rep. 1997;17:3-‐8.
4 Katia Aquilano et al. Role of Nitric Oxide Synthases in Parkinson’s Disease: A Review of the Antioxidant and Anti-‐inflammatory Activity of Polyphenols. Neurochem Res (2008) 33:2416-‐24263|Pageexcellent time (speaking of “number one suspect”) to suggest that the reason why man has been researching, looking for the cause and cure of diseases for centuries, yet can claim very few victories (don’t believe it? Start naming all of the cures man has discovered) is because researchers wear the researchers hat quite well, but then neglect to put on the detective or judge hat. Meaning, they seem to do quite well gathering up all the clues, then neglect to put them all together like a detective would do, and then make some bold assumptions to crawl out of the box and move forward like a judge would do. As you’ll see, we’ll be making bold, but intelligent assumptions as we move forward in this book.Let’s start with the question you must be asking yourself: Why hasn’t anyone blamed mercury up until now considering all the pain and misery it appears to have caused? For one thing, we know that mercury doesn’t hang around at the scene of the crime, but does it’s deadly deed, and moves on to imbed deep within the body’s bones and tissues. Without mercury hanging around to take the blame, it is clear why confusion exists even today, even with our modern means of testing.This appears to be what Professor Emeritus Martin L. Pall means when he talks about diseases sharing the same symptoms, each “initiated by a short-term stressor” only to be followed by chronic illness that…most often lasts for life.5 Mercury is a “short-term stressor” that has been confounding the minds of even the most brilliant men and women for centuries. Now that we have more abundant and free sharing of information let’s hope we can put together the puzzle pieces so we can solve the Parkinson’s mystery, once and for all.Again, we’ll see very clearly later why mercury specifically damages the mitochondria. But one indication that mitochondrial damage is part of Parkinson’s was discussed in a 1998 study: “Mitochondrial impairment as an early event in the process of apoptosis (cell death) induced by glutathione depletion in neuronal cells: Relevance to Parkinson’s disease”. They imply in the study that the glutathione depletion is the result of the mitochondrial impairment. In reality, glutathione transports mercury out of the body, and is also depleted by mercury,5 Martin L. Pall. A Common Causal (Etiologic) Mechanism for Chronic Fatigue Syndrome/Myalgic Encephalomyelitis, Multiple Chemical Sensitivity, Fibromyalgia and Post-‐Traumatic Stress Disorder. email@example.com|Page
7 A.A. Thaker and A.A. Haritos. Mercury bioaccumulation and effects on soluble peptides, proteins and enzymes in the hepatopancreas of the shrimp Callianassa tyrrhena. Comparative Biochemistry and Physiology Part C: Comparative Pharmacology. Vol 94, Issue 1. 1989, pp 199-‐205.7|Page
sulfhydryl groups are also called thiols and mercaptans. Interestingly, it was the ancient Romans who coined the term “mercaptan” because of sulfur’s ability to “capture” mercury.In macular degeneration and cataracts, researchers found that it is a sulfur compound that makes up the protein repair system in the function and maintenance of aging eyes.8 We’re going to find as we delve into this deeper, that the mitochondrial membrane, genes, enzymes, and antioxidants damaged by mercury, contain a sulfur group. So you can see how it is going to be difficult at this point to discuss damage to the mitochondria without mentioning mercury at nearly every turn. In the next chapter we go into much more detail how mercury is highly attracted to sulfur in the body.You might say that mercury is much like a nuclear explosion within the body, as mercury goes about damaging sulfhydryl groups wherever mercury shrapnel falls, because of that high affinity between the “soft” sulfide and the “soft” metal that is mercury. When mercury attaches to the sulfur, mitochondrial membranes are damaged, as are protective systems that contain sulfur, like glutathione.Man has needed to eliminate mercury as something we deliberately inject into people on a daily basis for centuries. The complete elimination of mercury from our environment and bodies would most likely end the one in one-hundred people getting Parkinson’s in America today (not to mention numerous other diseases). Would a random one in, say 10,000 still get Parkinson’s from extreme exposure to another toxicant like paraquat (or even living downwind of some factory spewing mercury into the air perhaps, or drinking water from pipes tainted by the lead used in soldering pipes)? Of course. The world is a toxic place, and exposure to enough of many toxins could cause neurological damage.8 Lisa A. Brennan, Marc Kantorow. Mitochondrial function and redox control in the aging eye: Role of MsrA and other repair systems in cataract and macular degenerations. Experimental Eye Research 88 (2009) 195-‐203.
Consider this, if your doctor told you that the immunization you were about to receive contained lead, would there not be an instant refusal to take the shot? How about if the shot contained arsenic? Of course you’d refuse! This is because it is common knowledge how toxic lead and arsenic are. In a 2010 series on CNN about toxins in America, it was explained how the Center For Disease Control (CDC) used to have a number they deemed a “safe” level of lead ingestion. The story is told of one man who set about to prove, via a town that was poisoned with lead from manufacturing, that the peoples’ sickness was worsened the closer they were to the plant. However, the sicknesses continued out in areas quite distant from the plant as well. He proved the point, and now the CDC acknowledges there is no safe level of lead. When will mercury be similarly acknowledged for the poison that it is? The truth is, the body has no beneficial use for lead or mercury. It is past time to brand mercury as the “no safe level” poison that it is so we can eliminate it from all medicines, dental materials, foodstuff, and God-willing, our environment once and for all.Interestingly, the myelin sheath, known to be damaged in multiple sclerosis, is made up in good part, of sulfhydryl groups (again, sulfhydryl groups are also called thiols).12 There is a strong connection between people having the thimerosal-containing (which is mostly mercury) hepatitis B injection and subsequently presenting with multiple sclerosis.13 A hepatitis B injection from a “multi-use” vial contains 12.5 mcg of mercury. The so-called “safe” level of mercury has been set at .1 mcg per 2.2 pounds of body weight per day, or about .32 mcg for a newborn. A 180 pound man, therefore, should not have more than 8.2 mcg of mercury in a day according to this fictitious “safe level”. Of course, if that adult also breathes the air, has amalgam fillings, and has fish for dinner, it’s easy to see how mercury is doing the damage that it is today.Researchers have observed that tumor necrosis factor alpha (TNFa) inhibits mitochondrial respirations (mitochondrial respiration occurs via those complexes I- IV previously mentioned). These same researchers note that TNFa appears to induce mitochondrial dysfunction. They attempted to figure out where, why and12 Thomas Weimbs and Wilhelm Stoffel. Proteolipid (PLP) of CNS Myelin: Positions of Free, Disulfide-‐Bonded, and Fatty Acid Thioester-‐Linked Cysteine Residues and Implications for the Membrane Topology of PLP.13 Miguel A. Hernan et al. Recombinant hepatitis B vaccine and the risk of multiple sclerosis. Neurology. 2004, 63:838-‐842.10 | P a g ehow the TNFa is generated. Nowhere in the study is mercury mentioned.14 Yet mercury causes the release of inflammatory cytokines (defined below) like tumor necrosis factor (TNFa). Mice exposed to mercury showed altered expression of TNFa as well as two other cytokines, interferon and interleukin-12.15In another study, low levels of mercury were put into drinking water for 14 days. The results showed that low levels of mercury caused lipopolysaccharide-induced p38 and extracellular signal-regulated kinase activation and downstream TNFa and Interleukin-6 expression.16 Excess TNFa expression is a phenomenon seen in ALS, MS, Parkinson’s, rheumatoid arthritis and other immune, degenerative and neurological conditions. Mercury has been shown to induce TNFa, deplete glutathione, increase glutamate and Ca2+ toxicity, all of which are involved in mitochondrial dysfunction, inflammation and the death of immune and neuronal cells.17,18TNFa is a cytokine (small protein secreted by the immune system) involved in inflammation. It’s main role is the regulation of immune cells. In Parkinson’s, the interest in TNFa is mostly in its ability to induce apoptotic neuron death. Therefore, it is the dysregulation of TNFa that has become of interest. Putting on my judge hat for a moment, it is obvious that focusing on TNFa is not getting to the core of the problem. TNFa doesn’t cause Parkinson’s, and more importantly, somehow “regulating” it by therapeutic means, won’t cure Parkinson’s. TNFa is a result of mitochondrial damage. Some Parkinson’s research focuses in on regulating TNFa as a possible way to control Parkinson’s. The truth is, stopping damage to and repairing the mitochondria is how we can stop the “dysregulation” of TNFa.14 J Stadler MD et al. Tumor Necrosis Factor Alpha Inhibits Hepatocyte Mitochondrial Respiration. Ann Surg (Nov 1992) 539-‐546.15 Sang Hyun Kim et al. Oral exposure to inorganic mercury alters T lymphocyte phenotypes and cytokine expression in BALB/c mice. Archives of Toxicology. Vol 77, No 11. 2003. 613-‐620.
16 Sang Hyun Kim, Sharma Raghubir P. Mercury alters endotoxin-‐induced inflammatory cytokine expression in liver: Differential roles of p38 and extracellular signal-‐regulated mitogen-‐activated protein kinases. Immunopharmacology and Immunotoxicology 2005 Vol 27 123-‐135.17 Noda M, Wataha JC, et al, Sublethal, 2-‐week exposures of dental material components alter TNFalpha secretion of THP-‐1 monocytes. Dent Mater. 2003 Mar;19(2):101-‐5
18 Dastych J, Metcalfe DD et al, Murine mast cells exposed to mercuric chloride release granule-‐ associated N-‐acetyl-‐beta-‐D-‐hexosaminidase and secrete IL-‐4 and TNFalpha. J Allergy Clin Immunol. 1999 Jun;103(6):1108-‐1411 | P a g e
So what is a kinase? Kinase is also known as a phosphotransferase and is a type of enzyme that transfers phosphate groups from high energy molecules such as adenosine triphosphate (ATP) to a specific substrate [a molecule upon which an enzyme acts]. This is called phosphorylation and is necessary to energize all that needs to occur in the body – like cell division, for one thing. (An enzyme that removes phosphate groups is known as phosphatase.) ATP is the molecule in the mitochondria responsible for transporting energy within cells for metabolism. ATP is produced by photophosphorylation, and by cellular respiration (via those mitochondrial complexes I-IV we keep mentioning). All to say, damaged mitochondria can’t do their job and the entire body suffers.Let me pause here and explain the reason for all the “science” in this book. If you read from beginning to end you should easily understand the four main points outlined at the very outset of the book. Whether or not you comprehend every single term is irrelevant. You will be able to make the proper conclusions, and from there you will be able to make wise decisions about yours or a loved-ones Parkinson’s. As doctors and medical professionals, it has always occurred to me that we don’t do patients any favors by leaving them in the dark about critical terminology, which includes the meaning of words that apply to their condition.All That Is Going On In Parkinson’s Emanates From The MitochondriaWhen we study mitochondrial function, we begin to see that nearly, or perhaps all, of “what is going on” in Parkinson’s (and many other neurological diseases) stem from mitochondrial damage, and most likely (or most often) by mercury. This is critical, because if we ever hope to prevent or cure Parkinson’s, the focus must be on protecting and repairing the mitochondria and this in turn would take care of “all the things going on”. A study from Cornell University states this about what causes degeneration of the nigrostriatal dopaminergic neurons in Parkinson’s disease: “…the role of mitochondrial dysfunction gains strongest support because mitochondria are central to a number of processes thought to be integral to PD pathophysiology.”20 (PD is the common abbreviation for Parkinson’s Disease).20 Bobby Thomas, PhD and M. Flint Beal, MD. Mitochondrial Therapies for Parkinson’s Disease. Movement Disorders Vol. 24, Suppl. 1, pp. S155-‐S160. 2010.
It has been discovered that neuronal mitochondria oxidize a combination of regular substrate pyruvate and glutamate, suggesting that mitochondria located at the inter-neuronal junctions, especially in the spinal cord, may be particularly vulnerable to oxidative stress.26GlutamateGlutamate is a neurotransmitter made from the amino acid L-Glutamine. Glutamate causes “excitation”. Where neurons synapse (connect) glutamate’s role is to stimulate the connection. In excess it is “excitotoxic”, literally causing neurons to fire wildly out of control, be damaged, even die. The healthy body makes all the glutamate it needs for proper synapsing, and all it needs in infinitesimal amounts.The amino acid L-glutamine is vital in many healthful functions in the body, including the good use of nitric oxide. In a healthy body, the following reaction is carried out by enzymes.Glutamate + ATP + NH3 → Glutamine + ADP + phosphate + H2O26 Alexander Panov MD, PhD Senior Scientist, Mitochondrial Biology Group, Carolinas Neuromuscular/ALS Research Laboratory, Department of Neurology. www.carolinasmedicalcenter.org
28 Angeles Almeida et al. Glutamate neurotoxicity is associated with nitric oxide-‐mediated mitochondrial dysfunction and glutathione depletion. Brain Research. Vol 790, Issues 1-‐2 (April 1998) 209-‐216.
discussed in more detail later), is inhibited by mercury (HgCl2) in vitro.29 In either case, what is being observed is a glutamatergic system not working properly.In addition, a deficiency of glutamine synthetase leads to an impairment of excitatory amino acid transporters (EAAT) which are found in neuronal and glial membranes.30 This results in the build-up of excess excitotoxic glutamate in astrocytes (a type of glial cell) as well as in the extracellular matrix, while simultaneously there is a decrease of glutamate inside neurons.31 Excess glutamate in the extracellular space over stimulates the NMDA receptor (a glutamate receptor).27,32,33,34 The over stimulation of the NMDA receptor results in alterations to calcium homeostasis.35 Influxes of calcium results in a change in membrane potential and the initiation of apoptosis, and ultimately, cell death.36For everyone, consuming free glutamates every single day is unhealthy to say the least. Glutamates added to foods is toxic by extreme excess, and something that must be dealt with. But for people with damaged mitochondria and resultant deficiencies in glutamate synthetase, consuming free glutamates adds tremendous insult to injury.When did we start adding free glutamates (which, in its pure form, is known as monosodium glutamate, or MSG) to our foods? Monosodium glutamate was discovered in 1908 by Kikunae Ikeda, a Japanese chemistry professor. For29 JW Allen et al. Mercuric chloride, but not methylmercury, inhibits glutamine synthetase activity in primary cultures of cortical astrocytes. Brain Res., 891, (2001)148-‐157.
30 Y Shigeri et al. Molecular pharmacology of glutamate transporters, EAATs and VGLUTs. Brain Res Brain Res Rev 45 (July 2004) 3:250-‐265.31 VA Fitsanaki & M Aschner. The importance of glutamate, glycine, and gammaaminobutyric acid transport and regulation in manganese, mercury and lead neurotoxicity. Toxicol Appl Pharmacol, 204, (2005)343-‐354.
32 JW Allen et al. The consequences of methylmercury exposure on interactive functions between astrocytes and neurons. Neurotoxicology, 23, (2002)755-‐759.33 E Mariussen & F Fonnum. The effect of polychlorinated biphenyls on the high affinity uptake of the neurotransmitters, dopamine, serotonin, glutamate and GABA into rat brain synaptosomes. Toxicology, 159, (2001)11-‐21.
34 E Matyja & J Albrecht. Ultrastructural evidence that mercuric chloride lowers the threshold for glutamate neurotoxicity in an organotypic culture of rat cerebellum. Neurosci Lett 158, (1993) 155-‐158. 35 DW Choi. Glutamate neurotoxicity in cortical cell culture is calcium dependent. Neurosci Lett 58, (1985) 293-‐297.36 TL Limke et al. Disruption of intraneuronal divalent cation regulation by ethylmercury: are specific targets involved in altered neuronal development and cytotoxicity in methylmercury poisoning? Neurotoxicology, 25, (2004)741-‐760.19 | P a g ecenturies Japanese cooks had been using various fermented food substances that when added to their cooking made food taste better. As Professor Ikeda discovered, the substance turned out to be L-glutamate. Monosodium glutamate was first marketed in 1909 as “Accent”. Since then, the evils of free glutamates have been uncovered as people suffered from what was at first described as the “Chinese Food Syndrome”. Now food manufacturers have become a bit sinister in their attempts to hide “MSG” by using various substances that contain free glutamate (see below). Today entire websites are maintained by people who suffer from the symptoms of excess free glutamates which include everything from severe migraines to neurological symptoms. It is said that over a million tons of free glutamate sources are sold and put into foods worldwide. Defenders of free glutamates just haven’t done their homework.MSG is just one source of free glutamates that most people have heard of. Most people think MSG is a preservative. But it’s not, it’s an “excitotoxin”, exciting the tastebuds, fooling them into thinking inferior food is far superior in taste than it actually is. The problem is that glutamates don’t just stimulate taste buds, but also causes excessive excitation of neurons in the brain and anywhere neurons are synapsing. Where there is damaged mitochondria, excess glutamate activity is already occurring because glutamate-clearing enzyme systems are damaged or missing. So ingesting glutamates in food becomes yet another neurological poison adding fuel to the fire.The worst part to the glutamate story is that food manufacturers hide free glutamates in up to 40 different additives so the consumer won’t know they’re ingesting it. Some names of compounds with hidden free glutamates are: yeast extract, hydrolyzed (anything), calcium caseinate, sodium caseinate, yeast food or yeast nutrients, autolyzed (anything), gelatin, textured protein, vetsin, ajinomoto, carrageenan, bouillons and broths, stock, whey protein, whey protein concentrate, whey protein isolate, any “flavors” or “flavoring”, maltodextrin, citric acid (E330), protease added, anything “enzyme modified”, malt extract, soy sauce, soy protein (unless it specifically says “whole soy”) and anything fermented. Often when something is “protein fortified”, beware. That protein is actually fermented, isolated, autolyzed, or textured, and thus full of free glutamates that are generated in the process.3737
Glutamate, like mercury is ubiquitous. Wherever you see a food with a long list of ingredients, many you can’t even pronounce, one or more of those ingredients is likely a free glutamate source. The daily ingestion of free glutamates is far in excess of what the body can handle. It isn’t surprising that we find glutamates being implicated in dozens of diseases, especially where damaged neurons are involved, as in Parkinson’s.38 Glutamates don’t cause the initial disease, but are being produced within the body excessively because of the disease in progress.Here’s an interesting thought. When you note that the Asian population seems far less affected by their extremely high level of free glutamate consumption, also note that they regularly drink sweet green tea, highest of all green teas in L-Theanine, a glutamate antagonist. There is some chatter about there being a synergistic benefit when L-Theanine is consumed in combination with caffeine (both of which are found in sweet green tea).Researchers have more recently found neurological disorders to be associated with a deficiency in glutamate dehydrogenase.39 Glutamate dehydrogenase is found in the mitochondria and is an enzyme that both breaks down and builds up L- glutamate. Like “all the other things going on” in Parkinson’s, the lack of glutamate dehydrogenase has been looked at as a possible cause of neurological diseases. However, when you put this finding into proper perspective, knowing that mitochondrial damage is the basis for Parkinson’s, you can see that a glutamate dehydrogenase deficiency is due to damaged mitochondria. Glutamate dehydrogenase deamination (breakdown) of glutamate requires NAD+ (nicotinamide adenine dinucleotide). NAD+ is also involved in the mitochondrial family of transport proteins, precisely where the mitochondria has been damaged.In Humans, the activity of glutamate dehydrogenase is controlled through ADP- ribosylation, a covalent modification carried out by the gene sirt4. This regulation is relaxed in response to caloric restriction and low blood glucose. It could very well be that the Ketogenic diet (high fat and protein, low carbohydrate) shown helpful against epileptic seizures and sometimes helpful for Parkinson’s has everything to do with the fact that blood glucose is kept low, increasing glutamate dehydrogenase, and consequently lowering excessive glutamate activity which fires neurons excessively (hence, a seizure or tremors). In fact, various modifications of the ketogenic diet, in attempts to make the diet less restrictive, but still creating a stable/low blood glucose have been shown to be as effective as the Ketogenic diet.40Another damaging effect of glutamate excitotoxicity is that it has been shown to lead to the excess production of nitric oxide. Glutamate excitotoxicity causes the rise of intracellular calcium which then increases neuronal nitric oxide synthase activity, the nitric oxide combines with superoxide anion to form peroxynitrite, and it is the peroxynitrite that has been shown as the main damaging molecule in dopaminergic neuronal cells. Thus, it is the production of large amounts of nitric oxide that are thought by many to contribute to dopaminergic neuron death.41 In case this is getting confusing, just remember. It all started with mercury damaging the sulfhydryl groups on the mitochondrial membrane (the various complexes that transport energy across the membrane). This in turn leads to a cascade of events that reduces antioxidants like glutathione, and increases excitotoxic glutamates and damaging nitric oxide.
As with so many compounds in the body, proper levels are good, but too much or too little is bad. As a messenger molecule nitric oxide is involved in many physiologic processes, including vasodilatation, immune response and neurotransmission.42 Proper levels of nitric oxide are necessary to keep blood pressure normal. Nitric oxide is only the beginning. It’s what happens when nitric oxide reacts with a dysfunctional mitochondrial electron transfer chain, and reactive oxygen species being spewed from the mitochondria that is of critical importance to the health of dopaminergic neurons.Nitric oxide is produced from the amino acid arginine via the enzymes inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase (nNOS). Only recently has an additional form of nitric oxide synthase been discovered in the mitochondria (mtNOS). This enzyme is currently being researched, but is thought to actually be iNOS, eNOS or nNOS translocated into mitochondria to participate in the regulation of the electron transfer chain.43,44 The electron transfer chain (ETC) is the part of the mitochondria we’ve previously mentioned as Complex I-IV (refer back to our chart at the beginning of the book)The study above says superoxide production occurs during dopamine metabolism as if somehow dopamine produces the damaging molecule. This may be an observation that is not fully descriptive of what really happens. Because we now know that when the mitochondrial membrane is damaged, there is long-term exposure of mitochondrial respiratory activities to nitric oxide which increases the excess production of superoxide anions (O2-), hydrogen peroxide and peroxynitrite. The excess production of these aforementioned reactive oxygen species results in a persistent inhibition of NADH:cytochrome c reductase activity (Complex III) which has been shown to inhibit Complex I.45 We also42 WK Alderton et al. Nitric oxide synthases: structure, function and inhibition. Biochem (2001) 357:593-‐ 615.
43 MC Carreras et al. Nitric oxide, complex I, and the modulation of mitochondrial reactive species in biology and disease. Mol Aspects Med (2004) 25:125-‐139.44 C Giulivi et al. Nitric oxide regulation of mitochondrial oxygen consumption I: cellular physiology. Am J Physiol Cell Physiol (2006) 291:C1225-‐C1231.
45 Riobo NA et al. Nitric oxide inhibits mitochondrial NADH:ubiquinone reductase activity through peroxynitrite formation. Biochem J. 2001 Oct 1;359(Pt1):139-‐45.23 | P a g eknow that glial cells produce excessive levels of nitric oxide in Parkinson’s disease which would also be neurotoxic for dopaminergic neurons.46Thus, one pathway leading to excess nitric oxide involves damage to mitochondrial Complex I activity of the electron transport chain (likely by mercury). There are other toxins that are actually used in labs to induce “parkinsonian” symptoms (discussed more later). Let’s just say here that you will see that these other toxins are now known to induce parkinsonianism by damaging the mitochondria.When complex I is damaged, impairing oxidative phosphorylation, we see an enhancement of excitotoxicity (excess glutamate and thus excess neuronal synapsing). Excitotoxicity leads to an influx of calcium, followed by activation of neuronal nitric oxide synthase. The excess nitric oxide combines with superoxide to form peroxynitrite and neurotoxicity ensues. During all of this, thiols (in the mitochondrial membrane as well as the antioxidants attempting to come to the rescue) react rapidly with a metabolite of nitric oxide to form S-nitrosothiols47 (another “reactive nitrogen species”, also known as thionitrites because they are a nitroso group attached to the sulfur atom of a thiol).We know that DNA damage occurs by direct reaction with reactive nitrogen species wherein repair processes are inhibited by lipid peroxidation products and/or hydrogen peroxide. The fragmenting of mitochondrial DNA can be attributed to reactive nitrogen species.48,49Protein modifications seen in Parkinson’s are caused by nitric oxide through nitrosylation and nitration. For example, during nitration, a nitro (-NO2) group is46 S. Hunot et al. Nitric oxide synthase and neuronal vulnerability in Parkinson’s disease. Neuroscience Vol 72, Issue 2. May 1996. 355-‐363.
47 Christina C. Dahm et al. Persistent S-‐Nitrosation of Complex I and Other Mitochondrial Membrane Proteins by S-‐Nitrosothiols but Not Nitric Oxide or Peroxynitrite: Implications for the interaction of nitric oxide with mitochondria. Journal of Biological Chemistry. Vol 281. No 15. April 14, 2006.48 PK Kim et al. The regulatory role of nitric oxide in apoptosis. Int Immunopharmacol (2001) 1:1421-‐ 1441.
49 Sten Orrenius et al. Mitochondria, oxidative stress and cell death. From “Oxidants And Antioxidants in Biology” Book of Abstracts. Translational Redox Science Co-‐Sponsored by the Linus Pauling Institute. Oxygen Club of California 2010. P. 2624 | P a g eadded onto tyrosine to form nitrotyrosine. Increased nitrotyrosine was detected in the substantia nigra of in vivo models of Parkinson’s.50In fact, increased nitrotyrosine was found in the core of Lewy bodies (abnormal aggregates of protein that develop inside nerve cells ) where a specific nitrated form of a-synuclein was also found. Of course, a-synuclein is also found to be highly expressed in the substantia nigra of Parkinson’s patients.51,52S-nitrosylation occurs when nitric oxide reacts with proteins through their reactive cysteine thiols.53 Modified proteins seen in neurological diseases like Parkinson’s include N-methyl-D-Aspartate receptor (NMDAR), p21ras, caspase 3 and 9, Nuclear Factor ӄB (NF-ӄB) and others.54In addition, increased levels of nitric oxide leads to specific S-nitrosylation of protein-disulphide isomerase (PDI). The up-regulation of PDI seems to be an adaptive response to protect neuronal cells, but S-nitrosylation of PDI leads to the accumulation of polyubiquitinated proteins leading to neuronal cell death via endoplasmic reticulum stress.55Nitric oxide has an affinity for heme (iron-containing molecule). Thus, nitrosylation or oxidation of protein thiols and removal of iron from iron-sulphur clusters in the mitochondria by nitric oxide will inhibit ATP synthesis.56 Some suggest that nitric oxide could actually be the primary compound involved in inhibiting complex I in dopaminergic neurons.57,5850 S Pennathur et al. Mass spectrometric quantification of 3-‐nitrotyrosine, orthotyrosine, and o,o’-‐ ditryrosine in brain tissue of 1-‐methyl-‐4-‐phenyl-‐1,2,3,6-‐tetrahydrophyridine-‐treated mice, a model of oxidative stress in Parkinson’s disease. J Biol Chem 274:34621-‐34628.
51 PF Good et al. Protein nitration in Parkinson’s disease. J Neuropathol Exp Neurol. (1998) 57:338-‐342. 52 BL Giasson et al. A hydrophobic stretch of 12 amino acid residues in the middle of a-‐synuclein is essential for filament assembly. J Biol Chem. (2001) 276:2380-‐2386.53 DT Hess et al. S-‐nitrosylation: spectrum and specificity. Nat Cell Biol (2001) 3:E46-‐E49.
54 DT Hess et al. Protein S-‐nitrosylation: purview and parameters. Nat Rev Mol Cell Biol (2005) 6:150-‐ 166.
55 T Uehara et al. S-‐nitrosylated protein-‐disulphide isomerase links protein misfolding to neurodegeneration. Nature (2006) 441:513-‐517.
56 S Moncada. Nitric oxide and cell respiration: physiology and pathology. Verh K Acad Geneeskd Belg (2000) 62:171-‐179 discussion 179-‐181.
57 AH Schapira. Etiology of Parkinson’s disease. Cell Death Differ (2007) 14:1261-‐1266.25 | P a g eAll of this, of course, occurs because mitochondria have been damaged by an initial stressor, most likely, most often mercury. It is important to keep the initial stressor in mind because just fighting nitric oxide won’t cure the underlying problem.In fact, because nitric oxide is being produced in excess and creating highly toxicmetabolites, this would indicate a need to inhibit excess nitric oxide, not attempt tostimulate it.59 Yet some studies recommend therapies or supplements to increasenitric oxide. Creatine and arginine would be examples of two supplements inpart aimed at increasing nitric oxide. Again, nitric oxide overproduction is whatresearch has shown to be the event significantly contributing to death ofdopaminergic neurons, via oxidative damage on cellular lipids, proteins and DNA.60,61A bit on the flip side, some researchers, in an attempt to protect good nitric oxide in the vasculature (eNOS), and in the immune and cardiovascular system (iNOS) have developed a class of “nonpeptide nNOS-selective inhibitors”. The thinking is that if they can keep nitric oxide down in the brain, but up everywhere else, they’d solve the problem. Of course, they are looking for a drug to do this. They still call it a “potential” therapy.62 But right off the bat, there’s a problem. It is inducible nitric oxide (iNOS) synthase that has been found in motor neuron mitochondria and Schwann cells, and thought to contribute to disease mechanisms in ALS.63 This once again reinforces the need to aim our “extinguisher” at the base of the fire, and not at “all the things going on”.58 MC Carreras et al. Nitric oxide, complex I, and the modulation of mitochondrial reactive species in biology and disease. Mol Aspects Med (2004) 25:125-‐139.
59 MF Beal. Excitotoxicity and nitric oxide in Parkinson’s disease pathogenesis. Ann Neurol. 1998;44:S110-‐114.60 JB Schulz et al. Inhibition of neuronal nitric oxide synthase by 7-‐nitroindazole protects against MPTP-‐ induced neurotoxicity in mice. J Neurochem (1995) 64:936-‐939.
61 P Hantraye et al. Inhibition of neuronal nitric oxide synthase prevents MPTP-‐induced parkinsonism in baboons. Nat Med (1996) 2:1017-‐1021.62 Richard B. Silverman. Design of Selective Neuronal Nitric Oxide Synthase Inhibitors for the Prevention and Treatment of Neurodegenerative Diseases. Accounts of Chemical Research. Vol 42. No 3. March 2009. 439-‐451.
63 Kevin Chen et al. Inducible nitric oxide synthase is present in motor neuron mitochondria and Schwann cells and contributes to disease mechanisms in ALS mice. Brain Struct Funct (2010) 214:219-‐ 234.26 | P a g eIn 2010 researchers Khan and Ghosh report on the herb Withania somnifera for possibly lowering nNOS.64 The benefit of looking to herbal therapies is that herbs tend to go where they are needed, and do their job without the dangerous side- effects of man-made drugs.65Another “herb” a type of edible mushroom contains polysaccharides that were found to prevent inflammation through the inhibition of both Cox-2 and nitric oxide production, dose dependently.66Compounds that inhibit nitric oxide biosynthesis were shown to significantly protect dopamine neurons against manganese chloride (a compound known to stimulate microglia to produce reactive oxygen species).67Indeed, the “good guy” “bad guy” nature of nitric oxide is a good example of why trying to control events far from the source of the problem can be highly frustrating, useless, and even detrimental. It would appear that neither stimulating nitric oxide nor making heroic efforts to squelch it, is the answer in Parkinson’s and other neurological diseases. Trying to regulate all the “things going on” can (and has) failed thus far. What would be safe and beneficial, however, is consuming copious plant polyphenols as antioxidants against the damaging effects of nitric oxide.First and foremost, however, the most important step that can be taken to stop the excess production of nitric oxide is to eliminate the consumption of all dietary free glutamates. It goes without saying, of course, that we should aim our efforts at truly repairing the mitochondria, while keeping mercury and any other poison from damaging it further. In this way all the toxic things going on as a result, like excess nitric oxide production, should cease.The Mitochondrial Connection64 Zaved Ahmed Khan and Asit Ranjan Ghosh. Possible nitric oxide modulation in protective effects of Withaferin A against stress induced neurobehavioural changes. Journal of Medicinal Plants Research Vol 4(6) 490-‐495. 18 March, 2010.
65 Alan Tillotson. Constituents and Tissue Affinities in Herbal Medicine. Journey of Dietary Supplements. Vol 5. Issue 3. November 2008. 238-‐247.66 Byung Ryong Lee et al. Agrocybe chaxingu polysaccharide prevent inflammation through the inhibition of COX-‐2 and NO production. BMB Reports. June, 2009. www.bmbreports.org
67 Ping Zhang et al. Microglia enhance manganese chloride-‐induced dopaminergic neurodegeneration: Role of free radical generation. Experimental Neurology 217 (2009) 219-‐230.27 | P a g eWithin the now hundreds of studies making the mitochondrial connection to neurological and immunological diseases, none thus far have clearly elucidate how the mitochondria got damaged in the first place. They all state the obvious, that oxidative damage both to and from the damaged mitochondria are “involved.” I’m sure you can see how we need to take this a step further!As we move forward, never lose sight of the fact that mitochondria are self- replicating units within the cells. Damaged mitochondria that don’t die altogether, will produce more damaged mitochondria. In 1999 Kowald observed the “slow accumulation of impaired mitochondria”.68 In 2009, Baqri et al observed that damage to mitochondria involves the disruption of mitochondrial DNA, but that this does not hinder mitochondria from replicating, i.e., damaged mitochondria produce damaged mitochondria.69Some say it is very difficult to study the mitochondria because when you isolate them for study in a lab, or view them in a sacrificed animal, they aren’t in their live active state. Or as one group of researchers put it: “as with all mitochondrial incubations, the environment and substrate availability depart considerably from the physiological.”70 Nevertheless, much has been learned. But again, what seems to elude researchers to this day is what would cause that initial defect in or damage to the mitochondria. We need to explore this question thoroughly, because until we are confident we have unearthed the initial stressor, we will continue to study “all the things going on” without understanding why.So what damages the mitochondria? Mercury must be our number one suspect today, given this distinction by virtue of its prevalence and deliberate injection and placement into bodies. Mercury is well known to damage the mitochondria.7168 Axel Kowald. The mitochondrial theory of aging: Do damaged mitochondria accumulate by delayed degradation? Experimental Gerontology. Vol 34, Issue 5. August 1999. 605-‐612.
69 Rehan M. Baqri et al. Disruption of Mitochondrial DNA Replication in Drosophila Increases Mitochondrial Fast Axonal Transport In Vivo. November 17, 2009. www.plosone.org70 Sung W. Choi et al. Bioenergetic analysis of isolated cerebrocortical nerve terminals on a microgram scale: Spare respiratory capacity and stochastic mitochondrial failure. J Neurochem 2009 May; 109 (4):1179-‐1191.
71 Pilar Carranza-‐Rosales et al. DMPS reverts morphologic and mitochondrial damage in OK cells exposed to toxic concentrations of HgCl2. Cell Biology and Toxicology. Vol. 23, Number 3/May, 2007 p 163-‐176.28 | P a g e
MERCURYWithin this chapter we will make optimal use of our detective and judging skills, to prove beyond a shadow of a doubt, that mercury is the number one suspect, causing devastating damage to mitochondria as well as protective systems within the body.Mercury was known to be neurotoxic in the time of Julius Caesar and the Roman Empire, 100-44 BC. It was well-known that people who worked in the mercury mines at that time (they mined mercury from cinnabar, an ore containing mercuric sulfide) went crazy, suffered neurological symptoms, even died within a few years. Indeed, we’ve proven from vials of mercury found in ancient caves, that the deadly substance has been harnessed and used since antiquity.72 What you will likely find amazing, as do I, is that over the centuries, man has never lost sight of the knowledge that mercury is highly toxic. Why is that amazing? Because we use it liberally in medicine, dentistry and industry, knowing it’s one of the most toxicthings on earth.In fact, mercury is currently third on the list of the Center for Disease Control’s most toxic substances in our environment.73 So why do we put even “trace” amounts ofmercury into anything? Especially into things that we inject or implant into our bodies (like immunizations and amalgam fillings). Yet, in 2010, mercury is still used in the medical field, as well as in “energy saving” lightbulbs, and in industry which spews it into our waterways, polluting seafood.72 Mercury -‐ element of the ancients. Center For Environmental Health Sciences. Dartmouth College. 73 http://www.atsdr.cdc.gov/cercla/07list.html30 | P a g eThe 2007 CERCLA table goes on to list a total of 275 toxic substances. You would certainly recognize many of them, like cadmium, DDT and chloroform. If you go to their website and take a look at the entire list you’ll find that aluminum, fluoride and other familiar substances are on it as well. But mercury is #3 only after lead and arsenic (would you ever expect a doctor to inject you with either of those?) I ask again, why can he inject patients with mercury with impunity?CERCLA (The Comprehensive Environmental Response, Compensation and Liability Act) is a government agency that establishes requirements for other agencies within the government, like the Environmental Protection Agency. The way substances are ranked, is based upon frequency of occurrence at sites studies, toxicity of the substance, and potential for human exposure. Considering all the confusion about what is causing our epidemic of neurological and other diseases, today, it would behoove us all to take a serious look at this list. Fluorine, for example, is way down near the bottom of the list at #211. As “detectives” shouldn’t we conclude from mercury’s #3 ranking versus fluorine’s #211 ranking that you are perhaps 208 times more likely on a daily basis to be poisoned by mercury than fluorine?Yet even healthcare professionals are confused, throwing up their hands and saying, there are hundreds of poisons that can cause neurological diseases. Or worse, are so confused, it becomes easier to believe that only “spontaneous internal combustion” causes neurological diseases. The truth is, experiments with 18 other heavy metals have not produced even one of the many biochemical and histological diagnostic signs seen in the neurological disease Alzheimer’s which shares many of the same biomarkers as Parkinson’s.74So why mercury and not lead or arsenic as our number one suspect? We’ve already mentioned that immunizations are laced with mercury, not lead or arsenic. But also, as has previously been mentioned , it all has to do with mercury’s extraordinary affinity to sulfur, as in sulfhydryl groups found abundantly in the brain and nerves. In fact, mercury has possibly the highest affinity to sulfur of any substance known. I spoke of a “shocker” before, as to how it was known before74 FDA Panel Submission Mercury and Neurotoxicology.31 | P a g e
Outside of the body, mercury is found as elemental (Hg2+), inorganic (ethylmercury or iHg) and organic (methyl mercury MeHg). Whatever form, the76 Norrby, L.J. (1991). “Why is mercury liquid? Or, why do relativistic effects not get into chemistry textbooks?”.Journal of Chemical Education 68: 11033 | P a g e
Note many electrons unpaired
toxicity of mercury once inside the body lies in how it binds to sulfhydryl groups on proteins in the body.77 Of course proteins make up virtually everything in thebody – your genes, cells, and tissues – so altering them alters their function.78,79,80,81,82Nevertheless, methyl mercury has been dubbed the most toxic because it is a vapor, easily absorbed through skin and gut, and it easily crosses the blood-brain barrier. This is the mercury that vaporizes off of dental amalgams every time you chew on something. On the other hand, liquid elemental mercury, or inorganic mercury is poorly absorbed, but once inside the brain is even more neurotoxic than methyl mercury through astrocyte dysfunction .83Tragically a chemistry professor at Dartmouth, Karen Wetterhahn, died in 1997 from just a couple of drops of dimethyl mercury on the latex-gloves she was wearing. She was poisoned in August of 1996, and died in June of 1997. The chain reaction events that occurred with the mercury poisoning only showed up as deadly by the time she was hospitalized nearly a year after the tragic event.84Even today there is much confusion as to whether there is a “safe” mercury versus a “toxic” mercury. It was once thought that the blood-brain barrier (a system of tight junctions around capillaries and exists only in the central nervous system to protect the brain from dangerous foreign molecules) prevents mercury from entering the brain and thus doing any damage there. (A foolish assumption77 http://www.atsdr.cdc.gov/toxprofiles/tp.asp?id=115&tid=24
78 AI Cabanero et al. Effect of animal feed enriched with Se and clays on Hg bioaccumulation in chickens:in vivo experimental study. J Agric Food Chem (2005) 53, 2125-‐2132.
79 LG Costa et al. Developmental neuropathology of environmental agents. Annu Rev Pharmacol Toxicol (2004) 44, 87-‐110.
80 VA Fitsanakis et al. The importance of glutamate, glycine, and gamma-‐aminobutyric acid transport and regulation in manganese, mercury and lead neurotoxicity. Toxicol Appl Pharmacol (2005) 204, 343-‐354. 81 J Gailer et al. Structural basis of the antagonism between inorganic mercury and selenium in mammals. Chemical Research in Toxicology, (2000)13, 1135-‐1142.
82 A Szasz et al. Effects of continuous low-‐dose exposure to organic and inorganic mercury during development on epileptogenicity in rats. Neurotoxicology (2002) 23, 197-‐206.
83 A Yasutakeet al. Induction by mercury compounds of brain metallothionein in rats: HgO exposure induces long-‐lived brain metallothionein. Arch Toxicol, 72, (1998)187-‐191.
84 Dartmouth Toxic Metals Research Program: A Tribute to Karen Wetterhahn. http://www.dartmouth.edu/~toxicmetal/HMKW.shtml34 | P a g econsidering the toxic history of mercury, but we’ll proceed.) What we now know is that the blood-brain barrier prevents a “charged” form of mercury (charged because it has extra electrons) from crossing into the brain. The truth is, there’s simply no such thing as a “non-toxic” form of mercury, because once mercury enters the body the enzyme catalase strips an electron or two from a charged form of mercury (mercury chloride), creating an uncharged form (elemental mercury vapor) that readily crosses the blood-brain barrier.85Once mercury is in the brain, catalase will contact the mercury and charge it again, and it will then be unable to exit the brain. Enter glutathione. The antioxidant glutathione is the main detoxifier of mercury, and is itself a thiol (sulfur-containing compound). About 80% of ingested mercury can potentially be excreted by attaching to glutathione and exiting through the bowel. Unfortunately, we’re seeing that nearly every disease, including Parkinson’s, involves glutathione deficiency. This would be because of being used up by mercury, but also many other toxic substances, like acetaminophen, as well as from a diet devoid in nutrients necessary to make glutathione. One nutrient critically needed is sulfur, which would most abundantly be obtained from a diet rich in cruciferous vegetables. Bottom line is that without sufficient detox methods, like glutathione, mercury is reabsorbed and continues doing damage.As we will discuss more later, simply taking glutathione orally or even intravenously doesn’t appear to solve the problem. In bodies with damaged mitochondria, and used up glutathione, there are elevated enzymes (attempting to compensate for the lack of glutathione) that break down much of the attempt to replenish glutathione. When glutathione breaks down, one of its component parts, glutamate is released and contributes to the problems already in progress. Also as discussed earlier, but it bears repeating, in a mercury-damaged brain, there is an inhibition of glutamine synthetase (GS).86 Normally, glutamine synthetase catalyzes the ATP-dependent condensation of glutamate with ammonia to yield85 E Duhr, B Haley et al. Hg2+ Induces GTP-‐Tubulin Interactions in Rat Brain Similar to Those Observed in Alzheimer’s Disease. Federation of American Societies for Experimental Biology (FESAB). 75th Annual Meeting. Atlanta, GA 21-‐25 April 1991. Abstract 493
86 JW Allen et al. Mercuric chloride, but not methylmercury, inhibits glutamine synthetase activity in primary cultures of cortical astrocytes. Brain Res (2001) 891, 148-‐157.35 | P a g eglutamine. In other words glutamine synthetase is crucial in recycling glutamate so it doesn’t build up and become “excitotoxic”. Lack of glutamine synthetase contributes to excess glutamate. Excess glutamate causes excess nitric oxide and resultant highly toxic metabolites like peroxynitrite and has been proven to result in the death of dopaminergic neurons.Glutathione needs to get back an electron it has lost to be reactivated, and it has been shown that vitamin C is critical in that process. A diet and supplement regimen rich in vitamin C, as well as sulfur-containing foods and supplements have been shown to elevate glutathione safely, abundantly, and better than taking glutathione itself. A caution here before we continue. There are supplements like whey protein (rich in sulfur proteins) that has been shown to elevate glutathione levels. Unfortunately the supplement is “whey protein isolate” which is a source of free glutamates, and therefore unsuitable for people with Parkinson’s.When damaged mitochondria spew out the reactive oxygen species hydrogen peroxide, the main antioxidant to protect neurons is catalase87 (it functions to “catalyze” hydrogen peroxide into water and oxygen).88 One molecule of catalase can convert millions of molecules of hydrogen peroxide to water and oxygen per second.89 Oral supplements are available for catalase as well some other antioxidants produced naturally within the body (e.g., SOD and glutathione). Unfortunately, taken orally, these antioxidants are merely broken down within the intestine before they ever reach the cells that need them. Thus, consuming foods rich in the building blocks required to make these natural antioxidants (manganese, zinc, copper, and selenium) is a more effective way to increasing their levels in the body. These trace elements are found abundantly in a diet made up of copious quantities of whole, organic plant foods.87 R Dringen et al. The glutathione system of peroxide detoxification is less efficient in neurons than in astroglial cells. J Neurochem (1999) 72, 2523-‐2530.
88 P Chelikani et al. Diversity of structures and properties among catalases Cell Mol Life Sci. 61 (January 2004) 2:192–20889 DS Goodsell “Catalase”. Molecule of the Month. RCSB Protein Data Bank. September 1, 2004.36 | P a g e
any maladies arising from such a toxicity. If a savvy dentist suspects mercury toxicity, he/she is supposed to refer the patient to their doctor or other healthcare professional. The irony here, is that a dentist who fully understands the toxicity of mercury would be the one who could safely remove amalgams, the source of the mercury. Without the dentist’s help, the patient is the one who is then left without a solution to their health dilemma.How Mercury Damages the MitochondriaA 2008 study lists many of the diseases known to have mitochondrial dysfunction at their core including diabetes, hyperlipidaemia, and hypertension. The researchers wonder “How does a mutation in mitochondrial DNA lead to disease at the cellular level, and how can a single mitochondrial DNA point mutation result in such a variety of renal and non-renal phenotypes…and why are some regions of the nephron seemingly more sensitive to mitochondrial dysfunction and damage by mitochondrial toxins?”91 Perhaps they would find their answer if they simply inserted mercury into the equation, that is, mercury’s extraordinary affinity to sulfur and the kidneys (rich in the aforementioned sulfur-rich microtubules).Why, again, mercury? Why not equally assume the toxin is a pesticide? One reason is that mercury does not break down into harmless substances, but repeatedly does damage, causing widespread devastation throughout the body (this damage is seen in mammals, plants, and insects). On the other hand, substances like rotenone (an “organic” pesticide used to induce parkinsonism in lab animals) does break down, quite easily, actually, by temperature, light oxygen and alkalinity. Rotenone also breaks down by the universal solvent, water, and by carbon.92 So you would have to deliberately encounter rotenone for it to be toxic.91 Hall AM et al. Renal function and mitochondrial cytopathy (MC): more questions than answers? QJM 2008 Oct;101(10):755-‐766.
92 http://myfwc.com/newsroom/Resources/News_Resources_PiranhaFAQs.htm38 | P a g eAs we discussed previously, some mercury can be eliminated from the body, bound to an antioxidant sulfur molecule (like glutathione), but the remaining mercury will come to “rest” deep within tissues, like in the bone.93Mercury does its dirty work and then flees the scene, and this is why researchers aren’t instantly pointing the finger at the poison. By the time damaged cells and tissues are studied, mercury is no longer necessarily hanging around at the scene of the crime – the obvious culprit. This leaves all the damage that was done to be studied and blamed.Why is this all so important? Because mercury’s devastating effects upon the “energy factories” of our bodies is not being given the weight of acknowledgement it merits, but is being “watered down” and lumped in with all the other potential dangers to the neurological system. Thus, people in the 21st century are still lining up for immunizations containing mercury. I cannot tell you how many times I’ve heard people say, “but they’ve taken the mercury out”. Wrong. Mercury (thimerosal) is still in “multi-use” vials in 2010. A sign seen during 2009’s “swine flu vaccination” campaign said: “If you have an allergy to thimerosal, let us know.” An allergy? To a poison they aren’t even honest enough to warn is really mercury?In fact, if you give birth to a baby in 2010, that infant is most likely going to be given a Hep B shot with 12.5 mcg of mercury the day it is born. Yet .1 mcg per 2.2 pounds of body weight is supposedly the “safe” limit per day.94 Watch closely, because if you blink your eyes in the birthing room you’ll miss it. It is often done by a nurse while the infant is in the warmer, minutes after birth!93 http://www.idph.state.il.us/envhealth/factsheets/mercuryhlthprof.htm 94 The Environmental Protection Agency (EPA).39 | P a g e
98 Sternlieb I, Goldfischer S. Heavy metals and lysosomes. Front Biol. 1976;45:185-‐200.42 | P a g e
100 Katalin Sas et al. Mitochondria, metabolic disturbances, oxidative stress and the kynurenine system, with focus on neurodegenerative disorders. Journal of the Neurological Sciences 257 (2007) 221-‐239. 101 Madia Trujillo and Rafael Radi. Peroxynitrite Reaction with the Reduced and the Oxidized Forms of Lipoic Acid: New Insights into the Reaction of Peroxynitrite with Thiols. Archives of Biochemistry and Biophysics (Jan 2002) Vol 397, Issue 1. 1 91-‐98.102 S Yee & BH Choi. Oxidative stress in neurotoxic effects of methylmercury poisoning. Neurotoxicology 17 (1996) 17-‐26.43 | P a g e
radicals (OH-), nitric oxide (NO), peroxynitrite (ONOO-) and hypochlorus acid (HOCl). These molecules can and do rapidly convert to even larger reactive intermediates that are even more dangerous and difficult to neutralize in the body. Again, this is absolutely critical, because if we’re going to stop the mess, we’re going to have to aim our therapies at the mitochondria where the damage first occurs, and the oxidative chain reaction begins!With what we know about mercury’s affinity for the mitochondrial membrane, can we at least make sure we don’t jump through hoops with mitochondrially-targeted therapies and then continue to offer our patients immunizations with mercury? Indeed, what do a 10 year old, a 30 year old (like Michael J Fox) and a 65+ year old all have in common? Regular exposure to mercury! For centuries people have aged without getting Parkinson’s. Debilitating neurological diseases are just not a part of the “Master Plan”, and neither is the deliberate ingestion of mercury.Reactive Oxygen Species From Damaged MitochondriaDamage to the mitochondria’s electron transport chain results in continuous oxidative stress. It is a vicious cycle, unable to be broken due to antioxidant systems that normally keep reactive oxygen species under control. Thus, the problem is the imbalance between production and removal of reactive oxygen species. Again, reactive oxygen species are defined as any species capable of independent existence that contains one or more unpaired electrons.103During normal physiological reactions occurring in a healthy body, reactive oxygen species such as O2-, nitric oxide, and hydrogen peroxide perform beneficial functions before being neutralized by antioxidants. It is when they are produced in abundance, unchecked, or transform into highly aggressive free radical species that extensive cellular damage occurs. One such aggressive free radical is produced when hydrogen peroxide converts to the highly reactive hydroxyl radical (OH-). This reaction is usually catalyzed by iron. Another aggressive oxidant is peroxynitrite (ONOO-). Peroxynitrite occurs in the body when O2- reacts with103 Halliwell B and Gutteridge J.M.C. Free radicals in biology and medicine. Oxford University Press, Oxford, New York. 2007.44 | P a g enitric oxide104, both of which are produced in abundance by damaged mitochondria.Oxidative stress induces cytotoxicity by damaging DNA, proteins and lipids. In DNA damage, strands break, crosslinkages between DNA-DNA or DNA-protein and errors in DNA replication occur, and can also lead to tumor formation.105 Oxidative stress is seen in Parkinson’s and other neurological diseases as oxidative damage occurs to proteins resulting in protein-protein cross-linking, fragmentation, and “folding” if the side chains of the molecule are affected.106All from mercury, you say? Could one single poison be affecting so many people today? Well, it just so happens that also during the aforementioned Roman Empire the people decided to make their entire waterways out of lead. They used lead for cups and cooking pots and for vessels to make their wine. Josef Eisinger107 estimated that a Roman consuming a liter of wine a day would consume about 20 mg of lead, which he said was more than enough to produce chronic lead poisoning. Ever heard of the “Fall of the Roman Empire”? What do you suppose would happen to a civilization wherein every single resident consumed lead on a daily basis? And yet to this day, this is debated! The infertility of the women that resulted from lead ingestion, for example, is often surmised to have been merely from a desire to not have any children. I suggest we continue with eyes wide open, and know that if everyone in a society ingests a known poison continuously, it can, indeed, poison everyone.So before you grow weary of the emphasis on mercury, let me perhaps amuse you with an analogy. Consider that there are many ways to “die in a crash”. It can be an airplane crash, a train crash, a boat crash or an automobile crash. And just as statistics show you are most likely to die in an automobile crash, we are finding104 P. Pacher et al. Nitric Oxide and Peroxynitrite: in Health and disease. Physiological Reviews 2007, Vol 87(1) 315-‐424.105 UA Boelsterli. Mechanistic Toxicology: The molecular basis of how chemicals disrupt biological targets (2003) New York: Taylor & Francis
106 PD Josephy. Molecular Toxicology (1996) New York: Oxford University Press
107 Josef Eisinger. “Lead in History and History in Lead” (1984) Nature Publishing Group.45 | P a g ethat mercury is the most likely vehicle causing the “crash” in neurological (and many other) diseases today.Mercury And The Brain’s Immune SystemLet’s talk a little bit about the cells that make up the brains immune system. Within the brain (in addition to neurons) are glial cells (also called neuroglia or glia). Glial cells support and protect the brain’s neurons. There are about three glial cells for every neuron in the gray matter of the brain (which is where neurons reside – compared to white matter which mostly contains myelinated axon tracts).108 Glial cells surround neurons and hold them in place, supply nutrients and oxygen to neurons, insulate one neuron from another, and orchestrate neurotransmission. Glial cells also act as the brain’s “immune system” by destroying pathogens and removing dead neurons.109Astrocytes (also known as astroglia) are star-shaped glial cells in the brain and spinal cord. They perform many unique functions of their own, including support of endothelial cells which form the blood-brain barrier. Because the job of the blood-brain barrier is to keep harmful or foreign molecules out of the brain, as you can see, it is an important part of the brain’s immune system.Research since the mid-1990s has shown that, similar to neurons, astrocytes release transmitters (called gliotransmitters) in a Ca2+-dependent manner. It is thought that astrocytes signal to neurons through Ca2+-dependent release of glutamate which has now made astrocyte research of even more interest to neuroscientists. 110Then there are the antioxidants that are designed to protect the brain from oxidative damage. Glutathione is the main antioxidant protecting the brain from reactive oxygen species. Glutathione is a thiol, i.e., an organosulfur compound (contains a sulfur-hydrogen bond). There needs to be enough of the amino acid cysteine to manufacture glutathione. Cysteine is also a thiol. Cysteine is considered a non- essential amino acid because it is synthesized within the healthy body if there is108 Dale Purves et al. Neuroscience 4th ed (2008) Sinauer Associates 15–16.
109 H Wolosker et al. d-‐ Amino acids in the brain: d-‐serine in neurotransmission and neurodegeneration. FEBS J (2008 Jul) 275(14):3514-‐26.
110 TA Fiacco et al. “Sorting out Astrocyte Physiology from Pharmacology”. Annu Rev Pharmaco. Toxicol (October 2008) 49:15146 | P a g eenough of the essential amino acid methionine. An essential amino acid means it must be obtained from the diet.Methionine synthesizes some other nutrients in the body like the amino acid taurine, the phospholipid lecithin, and phosphatidylcholine (a phospholipid that makes up healthy cell membranes). Healing foods that are rich in methionine are sesame seeds, brazil nuts, garbanzo beans, corn, almonds, lentils and brown rice. If you are looking to boost methionine levels in your diet, however, look to the raw nuts and seeds above the lentils or rice, unless you sprout them and eat them raw. The reason for this is that cooking alters nutrients. With proteins, I like to use an egg as my standard for judging proteins/amino acids. Consider the appearance of a raw egg versus a cooked egg. The difference in appearance has to do with the fact that when you cook the egg you coagulate the protein and oxidize the fats. The white turns from clear to opaque. The yolk turns from bright yellow to an ashen gray yellow, and smells of sulfur that was released by the cooking.Of interest, too, is that methionine is converted to S-adenosylmethionine (SAMe) by methionine adenosyltransferase. SAMe acts as a methyl-donor in many methyltransferase reactions in the body. Methyltransferase reactions are performed in wide variety of biological functions in the body from DNA methylation in an early embryo’s development to recycling amino acids like methionine. Along the pathway for methionine, homocysteine is formed. Homocysteine is actually a homologue (has a similar chemical formula) to cysteine, and is seen elevated in cardiovascular disease and Parkinson’s. Homocysteine can be recycled back into methionine, but to do so, the body uses Vitamin B12 (cobalamin)-related enzymes. In addition, there needs to be enough folic acid (Vitamin B9) and Pyridoxine (Vitamin B6).111 Trimethylglycine (a betaine) is also a methyl donor, and has been shown to reduce homocysteine levels in blood.112111 JW Miller et al. “Vitamin B-‐6 deficiency vs folate deficiency: comparison of responses to methionine loading in rats”. American Journal of Clinical Nutrition 59 (1994) (5):1033–1039.
112 DA Coen et al. “Homocysteine-‐lowering treatment: an overview”. Expert Opinion on Pharmacotherapy 2 (2001) (9):1449–1460.47 | P a g eHow does this all relate to the brain’s immune system? Remember, the immune system you are familiar with protects your cells from damage from foreign invaders, like parasites but also mercury. Likewise, it has been shown that astrocytes release glutathione to generate a dipeptide CysGly (cysteinyglycine) which is then used by neurons as the precursor for their glutathione synthesis. Glutathione, of course, is the main detoxifier of mercury in the brain. In fact, in one study incubating neurons with astroglial cells resulted in neuronal glutathione levels twice that of neurons incubated without astroglial cells.113 Astrocytes have been shown to be more resistant to cytotoxicity than neurons.114,115 Why are astrocytes more resistant to cytotoxicity than neurons? It has been shown that excess glutamate can impair the Na+-dependent cysteine transporters116 which would lead to deficiencies in glutathione in neurons, which would make neurons less capable of detoxifying reactive oxygen species by the glutathione pathway. Interestingly, chronic methyl mercury exposure in humans showed a loss of neurons, and a proliferation of astrocytes.117In his Master’s thesis exploring the demethylation of methylmercury in the central nervous system, Aaron Shapiro explains how inorganic mercury and methyl mercury are both toxic, just through different mechanisms. He outlines studies showing how inorganic mercury preferentially target astrocytes, while methyl mercury targets neurons. Because inorganic mercury does not penetrate the blood- brain barrier, he concludes that the presence of inorganic mercury in the brain after orally administered methyl mercury can only be explained by demethylation in situ. He points to studies that show that demethylation occurs in the brain, driven113 Ralph Dringen et al. Synthesis of the Antioxidant Glutathione in Neurons: Supply by Astrocytes of CysGly as Precursor for Neuronal Glutathione. The Journal of Neuroscience (Jan 15, 1999) 19(1):562-‐569.114 JW Allen et al. Mercuric chloride, but not methylmercury, inhibits glutamine synthetase activity in primary cultures of cortical astrocytes. Brain Res 891 (2001) 148-‐157.
115 R Dringen et al. The glutathione system of peroxide detoxification is less efficient in neurons than in astroglial cells. J Neurochem 72 (1999) 2523-‐2530.116 JW Allen et al. The consequences of methylmercury exposure on interactive functions between astrocytes and neurons. Neurotoxicology, 23, (2002)755-‐759.
117 K Eto et al. Differential diagnosis between organic and inorganic mercury poisoning in human cases— the pathologic point of view. Toxicol Pathol 27 (1999) 664-‐671.48 | P a g eby oxidative stress. He said it would be “reasonable to assume” that mitochondrial reactive oxygen formation contributes to methyl mercury demethylation.118As we mentioned, optimal levels of glutathione can be maintained in astrocytes under oxidative stress conditions if there are enough glutathione precursors (sulfur amino acids) and vitamin C.119Mercury And The Viral ConnectionMany diseases are shown to have a “viral component”, so much so, that researchers often assume it is the virus that caused the disease in the first place. The truth is, viruses live dormant in nerve ganglia. When an opportune time arrives, viruses leave the ganglia to travel along nerves to swoop in and invade damaged cells . Healthy cells build up defenses against viruses while damaged cells are completely vulnerable to infection. The entire purpose for a viruses’ existence is to replicate. When they move into cells to do this, they kill the cell in the process.Mercury damages cells, and has been shown to increase mortality due to viral infections.120 Mercury has been found repeatedly to accumulate in macrophages121,122 interfering with macrophage functions in the body that control viral replication leading to advanced viral infections.123Injected mercury shows up accumulated in organs, including organs of the immune system, which include the spleen, thymus, lymph nodes and bone118 Aaron Shapiro B.Sc., University of Guelph. Characterization of Methylmercury Demethylation In The Central Nervous System. (2005) Thesis Submitted in Partial Fulfillment Of The Requirement For The Degree Of Master Of Science In Interdisciplinary Studies. (January 2008) The University of Northern British Columbia.119 E O’Connor et al. Biosynthesis and maintenance of GSH in primary astrocyte cultures: role of L-‐cystine and ascorbate. Brain Research (1995) Vol 680 Issues 1-‐2. 157-‐163.120 LD Koller Methylmercury: effect on oncogenic and nononcogenic viruses in mice. Am J Vet Res 36, (1975)1501–1504.
121 YM Sin et al. Effect of lead on tissue deposition of mercury in mice. Bull Environ Contam
Toxicol 34, (1985) 438–445.122 M Christensen et al. Histochemical and functional evaluation of mercuric chloride toxicity in cultured macrophages. Prog Histochem Cytochem 23, (1991) 306–315.
123 S Ellerman et al. Effect of mercuric chloride on macrophage-‐mediated resistance mechanisms against infection with herpes simplex virus type 2. Toxicology 93, (1994)269–87.49 | P a g emarrow.124,125,126,127 In a study to estimate the toxicity of mercury, electron microscopy showed that the mercury accumulated mostly in the lysosomes of macrophages and endothelial cells. Their study showed that the bone marrow showed only a few grains of mercury, found in cells “with macrophage appearance”.128Lysosomes are organelles (recall that mitochondria are also organelles). Lysosomes contain enzymes (acid hydrolases) that disassemble waste materials and cellular debris within the cell, including other worn-out organelles, food particles, and engulfed viruses or bacteria (engulfed by macrophages).Some Other Toxins That Have Been Shown To Cause ParkinsonismContrast mercury, now, with other toxins that are said to produce Parkinson’s (used mostly in lab animals and injected directly into the animal) like paraquat, first synthesized in 1882, but not used commercially until 1961129; or amphetamine, which was first synthesized in 1887 but not really used until 1927130. It’s not that we discount these possible toxins, but that we be realistic about the epidemic of Parkinson’s, and just what toxin(s) Parkinson’s patients are most likely to be exposed to, and have in common, especially for all these years since the “Shaking Palsy” was first documented centuries ago. If you start to get confused, referred back to the CERCLA list, and how it is generated, especially the part about its potential for human exposure.124 M Mehra & KC Kanwar. Clearance of parenterally administered 203Hg from the mouse tissues. Jepto 5-‐4/5 (1984) 127-‐130.
125 JK Nicholson et al. Comparative distributions of zinc, cadmium and mercury in the tissues of experimental mice. Comp Biochem Physiol B 77 (1984) 249-‐256.126 F Planas-‐Bohne et al. The influence of administered mass on the subcellular distribution and binding of mercury in rat liver and kidney. Arch Toxicol 56 (1985) 242-‐246.
127 JB Nielsen & O Andersen. Disposition and retention of mercuric chloride in mice after oral and parental administration. J Trace Elem Electrolytes Health Dis 30 (1990) 167-‐180.128 M. Christensen. Histochemical localization of autometallographically detectable mercury in tissues of the immune system from mice exposed to mercuric chloride. Histochemical Journal 28 (1996) 217-‐225. 129 http://www.inchem.org/documents/ehc/ehc/ehc39.htm Paraquat and Diquat
130 L. Edeleanu. Uber einige Derivate der Phenylmethacrylsaure und der Phenylisobuttersaure. Ber Duetsch Chem Ges 1887;Vol 20:616.50 | P a g eThat said, many other potential toxins aren’t normally encountered to the degree mercury is, but all must be considered. Of course, you might be right in suspecting something other than just mercury if you live in Somerville, Texas, where nearly every single resident suffers everything from birth defects to bladder, pancreatic or brain cancer from a massive wood-treatment facility, which for more than 100 years has churned toxic chemicals into the atmosphere while manufacturing phone poles and bridge supports. But then, mercury has been used historically for preserving wood. Were they using it in Somerville?While we must first condemn mercury, and do so quickly, because it is deliberately being injected, implanted and eaten on a daily basis, we must also consider the other neurotoxins that have been shown to induce Parkinson’s as well.OHDA In “Tyrosine Hydroxylase Gene Transfected Hematopoietic Stem Cells in a rat model of Parkinson’s disease”, they say that dopamine levels were restored in46.6% and 33% of control. They say the transferred cells showed excellent survival rates in PD rat brains, and distant migration was observed. The toxin used to induce the Parkinson’s? The rats were injected with 8 micrograms of 6- OHDA.131 6-OHDA is oxidized dopamine.So would we observe the same success with a tyrosine hydroxylase gene transfected hematopoietic stem cell transplant in a human with Parkinson’s? So far it appears that Parkinson’s in humans involves widespread damage to mitochondria and resultant loss of glutathione along with the generation of reactive oxygen species that ultimately does to transplanted hematopoietic stem cells what normally occurs in the Parkinson’s patient, that is, damage to any new dopaminergic neurons.Before we go on, we need to discuss the mitochondrial electron transport chain (ETC). The ETC occurs in the mitochondrial membrane, and is what we’re now seeing is damaged leading to a cascade of events, all of which are seen in Parkinson’s. Complex I (NADH Dehydrogenase) is the first enzyme of the mitochondrial electron transport chain. Complex I translocates four protons per131 Shizhong Zhang et al. The Therapeutic Effects of Tyrosine Hydroxylase Gene Transfected Hematopoietic Stem Cells in a Rat Model of Parkinson’s Disease. Cell Mol. Neurobiol (2008) 28:529-‐543.51 | P a g e
Paraquat appears to contribute to the generation of reactive oxygen species via mitochondrial damage, imbalances or dysfunction. When O2 is present, paraquat generates superoxide in high amounts which is what accounts for its toxicity. In fact, it is due to this toxic level of reactive oxygen species that paraquat inhibits photosynthesis and Co2 fixation in plants, killing the plant.Some studies show that complex I of the respiratory chain is the main site of superoxide production by paraquat. Paraquat PQ2+ undergoes univalent reduction to generate the paraquat radical which then reacts rapidly with O2 to produce superoxide.133 Other studies point to Complex III as the potential site of paraquat’s toxic action. Whatever the case, paraquat exposure has been identified in epidemiological studies as well as in toxicant-based models of Parkinson’s, points to the importance of mitochondrial damage and reactive oxygen species as that which leads to dopaminergic cell death, and thus Parkinson’s.Paraquat rapidly depletes glutathione and protein thiols, induces lipid peroxidation and its cytotoxicity is related with the uncoupling of oxidative phosphorylation in the mitochondria.134 Superoxide dismutase (SOD) is capable of inhibiting the actions of paraquat.135MPTP used to induce Parkinson’s in studies with animals, has provided several insights on the potential role of mitochondrial complex I dysfunction in PDpathogenesis.136 In fact, there are other even more potent complex I inhibitors (paraquat, rotenone, pyridaben, and fenpyroximate) which, when administered at even low doses have been shown to produce symptoms like Parkinson’s, and lead133 Helena M. Cocheme and Michale P. Murphy. The uptake and interactions of the redox cycler Paraquat with mitochondria. Methods in Enzymology Vol 456. 2009. pp 395-‐417.
134 Carols M. Palmeira et al. Thiols metabolism is altered by the herbicide paraquat, dinoseb and 2,4-‐D: A study in isolated hepatocytes. Toxicology Letters Vol 81, Issues 2-‐3, 15 November 1995, Pages 115-‐123. 135 B.J. Day et al. A metalloporphyrin superoxide dismutase mimetic protects against paraquat-‐induced endothelial cell injury, in vitro. J Pharmacol Exp Ther 275, 1227-‐1232.
136 Przedborski S et al. MPTP as a mitochondrial neurotoxic model of Parkinson’s disease. J. Bioenerg Biomembr 2004;36:375-‐379.53 | P a g eto neurodegeneration.137,138 Many potential toxins, but one underlying issue: Damaged and thus dysfunctional mitochondria.The Acetogenin Family of Compounds are the most potent Complex I inhibitors. Some compounds in this family that the average person mightencounter are erythromycin A, Azithromycin, amphotericin, and tetracyclines – all are antibiotics or antifungals.Aerotoxic Syndrome There is also something few people know about called Aerotoxic Syndrome. Engine oil leaks fill the cabin air with fumes. One pilotwho experienced this firsthand relates, “During the descent my first officer complained he was feeling very sick. He needed to put his oxygen mask on. A couple of seconds later I felt so bad I was ready to throw up all over the instrument panel.” The pilots donned oxygen masks, but spent the next few minutes paralyzed. So what happened?With all modern aircraft the outside air is compressed and heated by the jet engines. The technical term for this is “bleed air”. But there are no filters in the system, therefore it is possible that the breathing air becomes contaminated with heated engine oil, especially when maintenance is an issue, or if a seal starts to leak or fail completely. This happens more often than airlines would like to admit. The cockpit and the pilots get 100 percent bleed air. Flight attendants and passengers in the cabin usually only get 40 to 60 percent of this air which is then recirculated. Pilots have become ill and are now unable to fly. Susan Michaelis is a former pilot for an Australia airline. She says this problem is a massive cover-up. She has written a book where she documents hundreds of cases.139Organophosphates Investigations have turned up the reason this is a problem. Oil residue from modern jet engines contain numerous chemicals.137 Betarbet R et al. Chronic systemic pesticide exposure reproduces features of Parkinson’s disease. Nat Neurosci 2000;3:1301-‐1306.
138 Derek A. Drechsel and Manisha Patel. Differential contribution of the mitochondrial respiratory chain complexes to reactive oxygen species production by redox cycling agents implicated in parkinsonism. ToxSci Advance Access Oxford University Press, September 18, 2009.139 Captain Susan Michaelis, Editor. Aviation Contaminated Air Reference Manual. 2007.54 | P a g eOrganophosphates used as anti-wear agents. Among the ingredients is a powerful neurotoxin, tricresyl phosphate, or TCP. There is up to 5% TCP in the special high performance oils. Professor Chistiaan van Netten is a toxicologist at the Canadian University of British Columbia, and an expert on TCP. He warns that TCP in humans interferes with the electrical conduction within the nervous system and consequently paralyzes people. In fact, in the 1920s, Jamaican ginger, a prohibited alcoholic beverage, was tainted by TCP. Many people suffered serious or fatal neurological damage.The way TCP causes neurological damage is by inhibiting the enzyme acetylcholinesterase leading to a buildup of acetylcholine in the synapses between neurons. This ultimately leads to hyperactivity and death of the neurons.140Karen Burns, a former stewardess, lost her health after just one flight. This was because the cabin of her plane was filled with TCP. Now, she and two of her coworkers suffer from serious nervous system effects, and Parkinson’s-like symptoms.140 M. Mumtazuddin Ahmed and P. Glees. Neurotoxicity of tricresylphosphate (TCP) in slow loris. ACTA Neuropathologica. (1971) Vol 19, No 2, 94-‐9855 | P a g eALL THE “THINGS GOING ON” IN PARKINSON’S TRACE BACK TO MITOCHONDRIAL DYSFUNCTIONIron And Copper – Causal Or A Result?Some researchers are looking at iron and copper as possible causal factors in Parkinson’s. But limiting the ingestion of iron and copper from whole foods likely does little to alter iron or copper “behaving badly” in the brains of healthy people. Similarly, the trace amounts found in a natural, whole plant foods diet are not something that can or should be avoided by people even with neurological diseases! Indeed, Nature has put iron and copper in trace amounts in food for a purpose. In fact, the brain is naturally rich in iron. It is reported that iron deficiency is the most prevalent nutritional problem in the world today, with up to 5 billion people affected.141However, taking iron supplements is a different story. Studies have shown an almost two-fold increase in Parkinson’s patient who took daily iron supplements.142 With what we now know about mitochondrial damage, the iron supplements did not cause Parkinson’s, of course, but can play a key role in the progression of the disease once in progress. This would be akin to the fact that systems are damaged to handle the body’s natural manufacture and handling of glutamate. Consuming free glutamates in the diet also play a key role in the progression of the disease.Undeniably, iron overload is seen in many neurological diseases, including Parkinson’s. Excess iron has been associated with brain lesions, also seen in neurological diseases. In addition, the excess iron has been associated with the toxicity of mercury and other metals in the brain.143 Parkinson’s disease, in fact, is141 John Beard. Iron Deficiency Alters Brain Development and Functioning. The Journal of Nutrition. 2003 Supplement.
142 KM Powers et al. Parkinson’s disease risks associated with dietary iron, manganese and other nutrient intakes. Neurology (2003) 60:1761-‐1766.143 Miyasaki K et al. Hemochromatosis associated with brain lesions -‐ a disorder of trace metal-‐binding proteins and/or polymers? J Neuropathol Exp Neurol. 1977 Nov;36(6):964-‐976.56 | P a g echaracterized by specific brain lesions (areas of damage) found in substantia nigra and other subcortical nuclei, namely Lewy bodies, made up of aggregates of alpha- synuclein. a-synuclein functions, in part, to regulate dopamine transporter activities.Recently, researchers have found that where there is accumulation of a-synuclein, there is a decrease in Complex I activity in the mitochondria in Parkinson’s disease brains.144In a 1986 study “Iron, A New Aid in the Treatment of Parkinson Patients” Birkmayer and Birkmayer state: “Intravenously applied iron – in form of a ferri- ferro-complex exhibited a considerable benefit for all (Parkinson’s) patients treated so far. They regained a remarkable mobility”. 145 This would suggest, of course, that iron is actually deficient because of a damaged mitochondria’s inability to “traffic” it properly.Of course metal ions do participate in oxidative stress seen in Parkinson’s and other neurodegenerative diseases. In fact, metal ion chelators have shown therapeutic value in ameliorating oxidative stress. The safest chelators, however, appear to be dietary chelators which have been shown to negate and even reverse the role of metal ions in oxidative stress.146 These dietary chelators are included in the supplements and diet sections.Indeed, the mitochondria is a “trafficker” of iron, or as some have put it, the mitochondria is not just about energy transduction, but it is also a focal point of iron metabolism.147 When the mitochondria is damaged, iron would be trafficked improperly, would elevate where it normally does not belong, and would144 Devi et al. Mitochondrial import and accumulation of a-‐synuclein impair complex I in human dopaminergic neuronal cultures and Parkinson disease brain. Journal of Biological Chemistry, 283, 9089-‐ 9100. 2008.
145 Birkmayer and Birkmayer. Iron, a New Aid in the Treatment of Parkinson Patients. J Neural Transm (1986) 67: 287-‐292
146 Theresa Hague et al. Dietary chelators as antioxidant enzyme mimetics: implications for dietary intervention in neurodegenerative diseases. Behavioural Pharmacology (2006) 17:425-‐430.
147 Richardson DR et al. Mitochondrial iron trafficking and the integration of iron metabolism between the mitochondrion and cytosol. Proc Natl Acad Sci USA. 2010 Jun 15;107(24):10775-‐82.57 | P a g eparticipate in oxidative reactions. Here again we can see that it is the underlying problem of mitochondrial damage leading to excess iron that needs to be addressed.Frataxin is a protein that in humans is encoded by the FXN gene. Frataxin is found in the mitochondrion. When frataxin homologue YFH1 is missing in yeast strains, there is an accumulation of iron in the mitochondria which then damages the mitochondria. 148When frataxin is mutated, there is iron overload. In deficiency of frataxin/YFH1, researchers have identified 14 proteins which are selectively oxidized as well as decreased superoxide dismutase activity, which promotes protein oxidative damage as seen in neurological diseases. The addition of copper and manganese to the culture medium restored SOD activity, preventing both oxidative damage and inactivation of magnesium-binding proteins. Recovery of mitochondrial enzymes required the addition of manganese, and cytosolic enzymes were recovered by adding copper. It is the reduced SOD activity that contributes to the toxic effects of iron accumulation.149In a 2009 study from the University of Washington, researchers report on the mutations and deletions in the mitochondrial genome saying the losses in stability correlate with a reduction in the mitochondrial membrane potential. They state that analysis of cells undergoing this instability showed a defect in iron-sulfur cluster biogenesis, which requires normal mitochondrial function.150 This is certainly not the only study to find that defects in the biogenesis of iron-sulfur clusters arise as a consequence of mitochondrial dysfunction, and that this increases genetic instability.148 Francoise Foury and Driss Talibi. Mitochondrial Control of Iron Homeostasis -‐ A Genome Wide Analysis of Gene Expression Yeast Frataxin-‐Deficient Strain. JBC Papers December 8, 2000. firstname.lastname@example.org
149 Irazusta V et al. Yeast frataxin mutants display decreased superoxide dismutase activity crucial to promote protein oxidative damage. Free Radic Biol Med 2010 Feb 1;48(3):411-‐420.150 Veatch JR et al. Mitochondrial dysfunction leads to nuclear genome instability via an iron-‐sulfur cluster defect. Cell 2009 Jun 26;137(7):1247-‐58.58 | P a g eA 2010 study was done to determine the mechanisms of iron exit from the “ferritin cage” in the mitochondria. Ferritin is a protein found inside of cells that stores iron and releases it as needed. The researchers describe how twenty-four subunits of ferritin combine to form a circular protein cage around up to 4,500 atoms of iron. They say that there are two models with regard to how iron exits the ferritin cage, and is reutilized. The first model has to do with the lysosomes degrading the protein ring causing iron to exit, and the second model has to do with “pores” at the junctures where the ferritin proteins join to create the ring, and iron being released through those pores. The study says that some of the fundamental functions of iron protein are still unclear. They say “…it is not known how cytosolic ferritin is degraded, how stored iron is released.” They do point to increased protein degradation, and that blocking protein degradation prevented iron mobilization from cytosolic ferritins. With regard to excess iron being released, we know that mercury degrades proteins. Nowhere in the study is mercury mentioned, however. With regard to a normal biological amount of iron being released, it appears lysosomes may very well do the job of controlled protein degradation in order for the iron to release.151So it isn’t that iron is a toxin like mercury to be avoided or eliminated at all costs. It is that damaged mitochondria leads to a “misdistribution” of iron. A 2010 study sums it up: “Iron concentrations can rise to toxic levels in mitochondria of excitable cells”. They say “iron chelation is probably inappropriate for disorders associated with misdistribution of iron within selected tissues or cells.”152Iron would most definitely appear to be a result of Parkinson’s, not a cause.Not Simply A Dopamine DeficiencyEarlier I said that Parkinson’s is no longer best defined as simply a dopamine “deficiency”. A damaged mitochondria results in toxic elements that cause151 Yinghui Zhang et al. Lysosomal Proteolysis Is the Primary Degradation Pathway for Cytosolic Ferritin and Cytosolic Ferritin Degradation Is Necessary for Iron Exit. Antioxidant & Redox Signaling (2010) Vol 13, No 7. 999-‐ 1009.152 Kakhlon O et al. Iron redistribution as a therapeutic strategy for treating diseases of localized iron accumulation. Can J Physiol Pharmacol. 2010 Mar;88(3):187-‐96.59 | P a g e
Prior to 1960 doctors didn’t even know that Parkinson’s had anything to do with dopamine. It was Ehringer and Hornykiewicz who described Parkinson’s disease as a “dopamine deficiency” disease154, and patients were given the first dose of Levodopa (L-dopa) in 1961.155 While drugs to increase dopamine production or block its oxidation are also often prescribed, L-dopa is the drug against which all other drugs are compared, said to be the most effective dopaminergic treatment for Parkinson’s aka The Gold Standard.But why L-dopa? Why not give dopamine? L-dopa can cross the blood brain barrier, and once in the brain is converted into dopamine. Conversely, dopamine doesn’t cross the blood-brain barrier, and if given as a drug would build up and be toxic to the rest of the body. In fact, Carbidopa is now routinely combined with L- dopa. Carbidopa is a drug that restricts L-dopa’s conversion to dopamine outside of the brain (to prevent peripheral toxicity). Carbidopa inhibits an enzyme (aromatic-L-amino acid decarboxylase) which is important in the conversion of L-dopa to dopamine, thus preventing L-dopa from becoming Dopamine prior to reaching the brain. Since Carbidopa cannot cross the blood-brain barrier, once L- dopa crosses, it can form dopamine unrestrained.Over the years L-dopa has been shown to have many drawbacks. The side-effect of nausea and vomiting, caused by L-dopamine’s conversion to Dopamine peripherally, is generally offset by use of Carbidopa. But this leaves the two most disconcerting side-effects of L-dopa use, which are motor complications that worsen year after year, and what is seen by researcher’s as L-dopa’s “potential to induce free radical-mediated damage and thereby induce and or accelerate nigral neuronal cell dysfunction and death.”156 I’d like to explore that statement, because with what we now know about mitochondrial damage, we should be able to put past observations into current perspective.154 H. Ehringer, O. Hornykiewicz. Distribution of noradrenaline and dopamine (3-‐hydroxytyramine) in the human brain and their behaviour in diseases of the extrapyramidal system. Klin Wochenschr (1960) 38:1236-‐1239.
155 W Birkmayer, O. Hornykiewicz. The L-‐3,4-‐dioxyphenylalanine (DOPA)-‐effect in Parkinson-‐akinesia. Wien Klin Wochenschr (1961) 73:787-‐788.156 CW Olanow et al. Levodopa in the treatment of Parkinson’s disease: current controversies. Mov Disord (2004) 19:997-‐1005.61 | P a g eMuch about dopamine toxicity was observed 40 or more years ago, and has not yet been “unlearned” by most who read those studies. I think we can show now that what we have blamed dopamine for doing all these years is not being done by dopamine, but to dopamine, by the reactive oxygen species generated from damaged mitochondria.Consider the next paragraph. Many of these observations are from studies done in the 1970’s and 80’s. Most are done in a lab (in vitro) not in humans or even “primates” (in vivo). These studies note that all of their observations are “poorly understood, inconclusive, that mechanisms have not been elucidated, and results are confounded by the in vitro environment being nothing like the in vivo environment”. These and more “disclaimers” continue even as recently as 2009.157 As you consider the next paragraph, note that if you insert that reactive oxygen species is being generated by damaged mitochondria as that which is actually oxidizing dopamine, causing it to become a part of the problem instead of the solution, much of the mystery is solved.Dopamine undergoes autoxidation, semiquinone formation and polymerization with the production of radical species.158,159 Dopamine can be metabolized by monoamine oxidase to produce hydrogen peroxide (H2O2)160 The H2O2 produced by dopamine, in the presence of iron (Fenton reaction) produces the highly reactive hydroxyl radical. Yet, you can find these in scientific literature as being associated with mitochondria as well.Non-physiological release of synaptic dopamine (such as when excitatory glutamate causes the release of dopamine161,162) is thought to play a major role in157 Arnar Astradsson et al. The Blood-‐brain barrier is intact after levodopa-‐induced dyskinesias in parkinsonian primates-‐Evidence from in vivo neuroimaging studies. Neurobiology of Disease 35 (2009) 348-‐351.
158 PG Jenner, DG Graham. Oxidative pathways for catecholamines in the genesis of neuromelanin and cytotoxic quinones. Mol Pharmacol (1978) 14:633-‐643.159 DC Tse et al. Potential oxidative pathways of brain catecholamines J Med Chem (1976) 19:37-‐40.
160 RN Adams et al. 6-‐Hydroxydopamine, a new oxidation mechanism. Eur J Pharmacol (1972) 17:287-‐ 292.
161 H. Mount et al. Glutamate Stimulation of 3H Dopamine Release from Dissociated Cell Cultures of Rat Ventral Mesencephalon. Journal of Neurochemistry Vol 52 (April 1989) Issue 4. 1300-‐1310.62 | P a g edyskinesia (diminishing ability to voluntarily move muscles, and an increasing presence of involuntary movements like tics and tremors).163 Researchers once suggested that the dyskinesia might have to do with a disrupted blood-brain barrier.164 But Astradsson et al found the blood-brain barrier in parkinsonian primates exhibiting L-dopa-induced dyskinesia to be intact.165In 2009 researchers found that calcium homeostasis is dysregulated in Parkinson’s patients with “L-Dopa-induced dyskinesias”. They found a depressed Ca2+ rise in response to mitogen-induced activation (which means a chemical substance encourages a cell to begin cell division). This defect was more pronounced in L- Dopa-induced dyskinesia patients. They conclude that “second messenger levels (like cAMP and free intracellular Ca2+) are altered in the peripheral blood lymphocytes of Parkinson’s patients treated with dopaminergic agents”, and this results in further alterations in Ca2+ homeostasis. 166Along with dopamine toxicity, researchers say they have observed mitochondrial dysfunction, specifically complex I deficiency.167 Here’s a case of the cart pulling the horse.On the one hand, research shows that L-dopa can act as a pro-oxidant at high levels, while conversely, at more normal levels, acts as an antioxidant, inducing the upregulation of glutathione and other neuroprotective molecules possibly because162 N.V. Kulagina et al. Glutamate regulates the spontaneous and evoked release of dopamine in the rat striatum. Neuroscience Vol 102 Issue 1 (January 2001) 121-‐128.
163 JA Obeso et al. Pathophysiology of levodopa-‐induced dyskinesias. Ann Neurol 47. (2000) S22-‐S32.
164 JE Westin et al. Endothelial proliferation and increased blood-‐brain barrier permeability in the basal ganglia in a rat model of 3,4-‐dihydroxyphenyl-‐L-‐alanine-‐induced dyskinesia. J Neurosci 26 (2006) 9448-‐ 9461.165 Arnar Astradsson et al. The blood-‐brain barrier is intact after levodopa-‐induced dyskinesias in parkinsonian primates-‐Evidence from in vivo neuroimaging studies. Neurobiology of Disease 34 (2009) 348-‐351.
166 Fabio Blandini MD et al. Calcium HOmeostasis is Dysregulated in Parkinsonian Patients with L-‐Dopa-‐ induced Dyskinesias. Clinical Neuropharmacology (May/June 2009) Vol 32, No 3. 133-‐139.167 AH Schapira et al. Mitochondrial complex I deficiency in Parkinson’s disease. Lancet (1989) 1:1269.63 | P a g ethe drug acts as a “minimal stressor”, enhancing the production of these protective molecules.168,169Perhaps most importantly is that more recent researchers have cautioned against placing much relevance upon observations of L-dopa toxicity in vitro culture because the culture is missing the high ascorbate found in tissues. Thus much of the in vitro evidence for a toxic effect by dopamine on neuronal cells may very well be “artifactual” and not the same as would be observed within the body.170 In addition, many studies that have demonstrated L-dopa toxicity in culture, used rather high concentrations of L-dopa, that is, >50μM/L compared to the typical 10- 20 μM/L given to patients, of which only about 12% actually shows up in the cerebrospinal fluid.171Scientists have noted that when glial cells (the brain’s “immune system”) and ascorbate have been added to cultures testing for L-dopa toxicity, because this becomes a scenario more like that found in the substantia nigra, L-dopa toxicity was significantly diminished or even abolished altogether!172,173It seems extreme measures have to be taken to induce L-dopa toxicity in rats. High levels of L-dopa were injected into them in the presence of iron to cause toxicity.174168 C Mytilineou et al. Toxic and protective effects of Levodopa on mesencephalic cell cultures. J Neurochem (1993) 61:1470-‐1478.
169 MA Mena et al. Neurotrophic effects of L-‐dopa in postnatal midbrain dopamine neuron/cortical astrocyte cocultures. J Neurochem (1997) 69:1398-‐1408.
170 MV Clement et al. The cytotoxicity of dopamine may be an artefact of cell culture. J Neurochem (2002) 81:414-‐421.
171 CW Olanow et al. Temporal relationships between plasma and cerebrospinal fluid pharmacokinetics of levodopa and clinical effect in Parkinson’s disease. Ann Neurol (1991) 29:556-‐559.
172 MA Mena et al. Glia protect fetal midbrain dopamine neurons in culture from L-‐dopa toxicity through multiple mechanisms. J Neural Transm (1997) 104:317-‐328.
173 C. Mytilineou et al. Levodopa is toxic to dopamine neurons in an in vitro but not an in vivo model of oxidative stress. J Pham Exp Ther (2003) 304:792-‐800.
174 H Maharaj et al. Levodopa administration enhances 6-‐hydroxydopamine generation. Brain Res (2005) 1063:180-‐186.64 | P a g e
176 TL Perry et al. Nigrostriatal Dopaminergic neurons remain undamaged in rats given high doses of Levodopa and carbidopa chronically. J Neurochem (1984) 43:990-‐993.65 | P a g e
“Complex I” is NADH dehydrogenase (also referred to as NADH: quinone reductase) is an enzyme located in the inner mitochondrial membrane. NADH is used to catalyze the transfer of electrons from NADH to Coenzyme Q. Complex I produces superoxide as well as hydrogen peroxide. Both of these free radicals are seen in excess in Parkinson’s. Complex I is the “entry enzyme” of oxidative phosphorylation in the mitochondria. Phosphorylation is the addition of a phosphate group (PO4) to protein enzymes to activate them (via enzymes called kinases) or deactivate them (via enzymes called phosphatases).Calcium HomeostasisCalcium Homeostasis is the term for when the body is maintaining adequate calcium levels. But it is also used to denote a proper balance of calcium within cells and outside of cells. A proper balance of calcium flowing into and out of cells would indicate homeostasis of calcium. Excess calcium flowing into cells would be called in influx, and excess calcium flowing out of cells would be called an efflux.Ca2+ (calcium ion) efflux is seen in Parkinson’s. Many substances were tested on heart mitochondria to see which increased Ca2+ efflux the most. Of all the substances tested, methyl mercuric chloride was the most effective since it was active at ratios of about 1 nmol/mg of mitochondrial protein.177Acetylcholine receptors found in the plasma membrane of certain neurons as well as other cells (muscarinic ACh receptors) when incubated with mercury in vitro caused calcium release from intracellular stores.178Mercury-induced damage has been observed to organelles which store calcium (e.g., the smooth endoplasmic reticulum and the mitochondrion) which led to increased intracellular calcium levels.179177 EJ Harris and H Baum. Production of thiol groups and retention of calcium ions by cardiac mitochondria. Biochem J. 1980 March 15;186(3):725-‐732.
178 TL Limke et al. Disruption of intraneuronal divalent cation regulation by methylmercury: are specific targets involved in altered neuronal development and cytotoxicity in methylmercury poisoning? Neurotoxicology (2004) 25 741-‐760.66 | P a g eMercury inhibits sarcoplasmic ATPases (a class of enzymes that catalyze the decomposition of adenosine triphosphate) resulting in an altered calcium homeostasis.180 The sarcoplasm is the cytoplasm (the part of a cell that is enclosed within the cell membrane) of a muscle fiber and has a high concentration of calcium used for muscle contractions.In excitable cells like neurons, the mitochondrial membrane contains a protein (NCX) that removes calcium from cells (one ion in exchange for 3 ions of sodium). Recent studies suggest that “supraphysiological activation” of NCX contributes to neuronal cell death and reduces the ability to reestablish normal ionic homeostasis.181 Supraphysiological activation just means that something out of the ordinary has occurred outside of normal physiological body functioning to cause activation of NCX. Of course toxins, and mercury is the primary suspect, would be just that, out of the ordinary.Researchers in 1997 found a Parkinson’s disease link to defects in mitochondrial function. When they bypassed complex I with succinate, they were able to partially restore suppressed recovery rates of cytosolic calcium increase. They concluded that the “subtle alteration in calcium homeostasis of Parkinson’s disease cybrids (mitochondrially transformed cells) may reflect an increased susceptibility to cell death under circumstances not ordinarily toxic.”182How Does Increased Intracellular Calcium Lead To Toxicity?It is known that the removal of extracellular calcium increases survival rate of neurons in vitro.183 Elevated calcium levels lead to cell death either by apoptotic or necrotic pathways. Calcium activates the protease calpain (a protein belonging to the family of calcium-dependent, non-lysosomal cysteine proteases). When179 AP Somlyo et al. Calcium content of mitochondria and endoplasmic reticulum in liver frozen rapidly in vivo. Nature (1985) 314, 622-‐625.
180 JJ Abramson et al. Heavy metals induce rapid calcium release from sarcoplasmic reticulum vesicles isolated from skeletal muscle. Proc Natl Acad Sci U.S.A, (1983) 80 1526-‐1530.181 P. Castaldo et al. Role of the mitochondrial sodium/calcium exchanger in neuronal physiology and in the pathogenesis of neurological diseases. Progress in Neurobiology 87 (2009) 58-‐79. Italy.
182 JP Sheehan et al. Altered Calcium Homeostasis in Cells Transformed by Mitochondria from Individuals with Parkinson’s Disease. Journal of Neurochemistry (March 1997) Vol 68, No 3. 1221-‐1233.183 DW Choi. Glutamate neurotoxicity in cortical cell culture is calcium dependent. Neurosci Lett (1985) 58, 293-‐297.67 | P a g ecalpain is abnormally activated, it leads to cleavage of vital proteins which results in damage to and death of cytoskeleton (cellular “scaffolding” or “skeleton” made of protein contained within the cytoplasm [interior of cells]).184Calcium-activation of phospholipase A2 (an enzyme involved in fatty acid metabolism in cell membranes) disrupts membrane stability which ultimately leads to death of the cell.185 In normal brains cells, phospholipase A2 regulates the balance between inflammatory and antiinflammatory fatty acids in cell membranes. Increased phospholipase A2 is associated with many inflammatory conditions, including neurological diseases.N-methyl-D-aspartic acid (NMDA) is an amino acid derivative which acts as a specific agonist at the NMDA receptor mimicking the action of glutamate. NMDA receptors are a type of glutamate receptor where upregulation has been seen in neuronal injury. Researchers in 2005 discuss how in previous studies, NMDA antagonists failed to treat stroke/neurotrauma patients. Newer studies indicate it would have been because the time period in which initial NMDA receptor up- regulation occurs is critical. Only when NMDA antagonists are employed during extracellular calcium reperfusion neurotoxicity can be abated.186Genes, Genetic Mutations, Protein MutationsThe so-called “familial Parkinson’s” because of pathogenic mutations in genes such as a-synuclein, parkin, DJ-1, PINK1, LRRK2, and HtrA1 has been shown to either directly or indirectly link their pathogenic roles with mitochondrial dysfunction.187,188 Mice with increased mitochondrial accumulation of human a- synuclein developed significant mtDNA damage and impaired cytochrome oxidase184 KK Wang. Calpain and caspase: can you tell the difference? Trends Neurosci (2000) 23, 20-‐26.
185 UA Boelsterli. Mechanistic Toxicology: The molecular basis of how chemicals disrupt biological targets. New York: Taylor & Francis. (2003)
186 Xin Wen-‐Kuan et al. The removal of extracellular calcium: a novel mechanism underlying the recruitment of N-‐methyl-‐D-‐aspartate (NMDA) receptors in neurotoxicity. European Journal of Neuroscience (February 2005) Vol 21, No 3. 622-‐636.187 Thomas B, Beal MF. Parkinson’s disease. Hum Mol Genet 2007;16 (Spec No. 2): R183-‐R194.
188 Konstanze F et al. Mitonchondrial dysfunction in Parkinson’s disease. Biochimica et Biophysica Acta 1802 (2010) 29-‐44.68 | P a g e(Complex IV) activity which led to mitochondrial dysfunction. The researchers reported that the mitochondrial dysfunction led to increased susceptibility to neurodegeneration induced by mitochondrial toxins.27 Indeed, many studies have now shown a direct link between parkin and its role in mitochondrial function, especially its ability to interact with mitochondrial transcription factor A (Tfam) to enhance mitochondrial biogenesis.189 Studies with mice and a conditional knockout of transcription factor A caused progressive loss of nigrostriatal dopaminergic neurons as seen in Parkinson’s.190Because mercury is a widespread contaminant, researchers deliberately fed zebrafish a mercury-contaminated diet for 25 days to see the effects on gene expression.It is also interesting to note that researchers have identified a “subset” of individuals who they say are exceptionally vulnerable to the toxic effects of mercury. As I go on to explain why, also consider that it has been shown that people who get flu shots (with mercury) year after year are many-fold more likely to get Alzheimer’s later in life than those who do not get flu shots.That said, people who have been identified with the genotype APOe 4/4 are considered at risk for developing Alzheimer’s. The APOe 4/4 gene has two binding sites that contain arginine, whose job it is to remove excess cholesterol from the brain. There is also an APOe 2/2 gene, not prevalent in the brains of those who are genotype APOe 4/4. APOe 2/2 gene makes a transport vehicle that has four binding sites that contain sulfur. The sulfur, as we’ve discussed, has an extraordinary affinity for mercury, and is thus protective, as a vehicle to transport mercury out of the brain. The arginine in the APOe 4/4 gene does a wonderful job of transporting cholesterol, but it has no affinity for mercury. APOe 2/2 gene is protective against AD and APOe 4/4 is predictive of early onset. Of course, if mercury weren’t being ingested, it is likely that even the APOe 4/4 individual would not succumb to Alzheimer’s or any other neurological disease. And so it is,189 Kuroda Y et al. Parkin enhances mitochondrial biogenesis in proliferating cells. Hum Mol Genet 2006;15:883-‐895.
190 Ekstrand MI et al. Progressive parkinsonism in mice with respiratory-‐chain-‐deficient dopamine neurons. Proc Natl Acad Sci USA 2007;104:1325-‐1330.69 | P a g e
192 WD Ehman et al. Application of Neutron Activation analysis to the Study of Age Related Neurological Diseases. Biol Trace Elem Res (1987) Vol 5. 19-‐33.
193 Shastry BS. Neurodegenerative disorders of protein aggregation. Neurochem Int 2003;43:1-‐7.70 | P a g e
neutrophils.194 Integrins are receptors that mediate attachment between a cell and the tissues surrounding it.The inflammatory response is part of a healthy immune system, acting to counteract infections and injury. It is only when inflammation is excessive or chronic that we see neurodegeneration. The microglia are associated with inflammation. Microglia are the “resident macrophages” of the brain and spinal cord. Microglia constantly scavenge the central nervous system cleaning up plaques, infectious agents and damaged or dead neurons. To induce Parkinson’s in lab animals, scientists inject the animal with substances that activate microglia. Some of these substances are rotenone, paraquat, 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Because microglia initiate the inflammatory response, if the activation of microglia is continuous, the inflammation will be excessive or chronic (as happens when toxic damage has occurred) and dopaminergic neurons will be damaged or killed as seen in Parkinson’s.195Neurons Firing Wildly Out of ControlIt’s been shown that restless leg syndrome is from a “neurological firestorm” which refers to neurons firing wildly out of control. Migraines are also the result of a neurological firestorm. Likely all aberrant movement and pain within neurological diseases can be labeled as a result of a neurological firestorm. This is critical to bring to the forefront, because involved in neurological firestorms are excessive glutamate and nitric oxide release. Neurons fire wildly out of control because of excessive glutamate release at the synapse, and then nitric oxide’s excessive creation and/or attempt to come to the rescue. Which leads us to the reason most/all Parkinson’s patients should likely not be given glutathione. As we have seen, damage to mitochondria, glutathione and glutamate dehydrogenase in the bodies of people with Parkinson’s and other neurological diseases, prevents the proper use of glutathione, and so it breaks down and releases free glutamates.Glutathione Deficiency194 RG Worth et al. Mercury inhibition of neutrophil activity: evidence of aberrant cellular signalling and incoherent cellular metabolism. Scand J Immunol (2001 Jan) 53(1):49-‐55.
195 PS Whitton. Inflammation as a causative factor in the aetiology of Parkinson’s disease. Br J Pharmacol (2007) 150:963-‐976.71 | P a g eThe main reason there is that deficiency of glutathione is discussed in this study: “Mercury can accumulate in mitochondria because of their high affinity for binding thiols (sulfur-containing molecules), leading to the depletion of mitochondrial glutathione.”196Apparently giving oral glutathione is not the best method of boosting glutathione levels anyway. In a study by Reddy et al called The systemic availability of oral glutathione, he states: When seven healthy people were given a single application of up to 3,000 mg of glutathione, there was no increase in blood glutathione levels. The human gastrointestinal tract contains significant amounts of an enzyme (gamma-glutamyltranspeptidase) that breaks down glutathione.197 I reiterate that glutathione, broken down, generates “free glutamates” which have been implicated in numerous diseases as a damaging factor, including Parkinson’s.One way to increase glutathione safely is stated in this study: “…in one trial, blood glutathione levels rose nearly 50% in healthy people taking 500 mg. of vitamin C per day for only two weeks.”198 Another way is to consume copious quantities of sulfur-containing food.199Up-Regulation of Gamma-Glutamyltranspeptidase HomocysteineOne extremely significant adverse effect observed along with L-dopa therapy (and again, most likely due to mitochondrial production of free radicals, including oxidized dopamine) is the increase in total homocysteine in the blood which has been linked to affective/cognitive impairment, dyskinesia and vascular disease in patients with Parkinson’s.200,201,202,203196 Charles R Arthur et al Parkinson’s Disease Brain Mitochondria Have Impaired Respirasome Assembly, Age-‐Related Increases in Distribution of Oxidative Damage to mtDNA and No Differences in Heteroplasmic mtDNA Mutation Abundance. Molecular Neurodegeneration 2009 4:37.
197 Witschi A. Reddy et al. The systemic availability of oral glutathione. Eur J Clin Pharmacol 1992. 43:667-‐9.198 Am J Clin Nutr 1993;58:103-‐5.
199 Sulfur amino acid deficiency depresses brain glutathione concentration. Nutr Neurosci 2001;4(3):213-‐ 22.
200 P.E. O’Suilleabhain et al. Elevated plasma homocysteine level in patients with Parkinson disease: motor, affective and cognitive associations. Arch Neurol (2004) 61:865-‐868.72 | P a g e
202 S Zoccolella et al. Elevated plasma homocysteine levels in L-‐dopa-‐treated Parkinson’s disease patients with dyskinesias. Clin Chem Lab Med (2006) 44:863-‐866.203 Martin-‐Fernandez JJ et al. Homocysteine and cognitive impairment in Parkinson’s disease. Rev Neurol (Feb 1-‐15, 2010) 50(3):145-‐151.
204 Daniel Offen et al. Prevention of Dopamine-‐Induced Cell Death by Thiol Antioxidants: Possible Implications for Treatment of Parkinson’s Disease. Experimental Neurology 141, (1995) 32-‐39.73 | P a g e
But the human body isn’t a Petri dish or lab animal. First you need to know that glutathione is made up of the amino acids L-cysteine, L-glutamate and glycine. The liver & lungs are where most glutathione is made. Glycine & glutamic acid are plentiful within the body, so it is the availability of cysteine that controls how much glutathione you can make.When human subjects were given 3,000 mg of glutathione there was no increase in blood glutathione levels because the human gastrointestinal tracts contacts gamma-glutamyltranspeptidase (GGT) which breaks down glutathione.205 This generates free glutamates, which is toxic, and already elevated in Parkinson’s. But GGT is also found up-regulated in the substantia nigra of Parkinson’s (who have depleted glutathione) thought to be a “sparing” act of the brain, in response to depleted glutathione.206 Giving intravenous glutathione could therefore be contrary to good medical practice for Parkinson’s, because the GGT would break it down in the brain causing excess free glutamates and further excitotoxic activity.So what about supplementing NAC? It would seem to be the miracle everyone depleted of glutathione would need. Unfortunately, taking it as a supplement is a double-edged sword in people with mitochondrial damage. When the brain contains (as it does in Parkinson’s) high levels of homocysteine, add more cysteine, and you actually cause oxidative stress in the presence of iron and copper (redox- active transition metal ions).207,208,209 In addition, taking NAC, which would result in an increase in glutathione, would only be broken down by GGT in the Parkinson brain, generating excess glutamate. Excess glutamate, in addition to causing excitotoxic neuronal death, causes excess nitric oxide, a potent vasodilator which leads to inflammation and headaches.205 A Witschi et al. The systemic availability of oral glutathione. Eur J Clin Pharmacol (1992) 43:667-‐669. 206 SJ Chinta et al. Up-‐regulation of gamma-‐glutamyltranspeptidase activity following glutathione depletion has a compensatory rather than an inhibitory effect on mitochondrial complex I activity: implications for Parkinson’s disease. Free Radic Biol Med (2006) 40(9):1557-‐1563.207 YZ Tang et al. Free radicals, antioxidants and nutrition. Nutrition (2002) 18:872-‐879.
208 RA Patterson et al. Mechanisms by which cysteine can inhibit or promote the oxidation of low density lipoprotein by copper. Atherosclerosis (2003) 169:87-‐94.
209 P Munoz et al. Differences between cysteine and homocysteine in the induction of deoxyribose degradation and DNA damages. Free Radic Biol Med (2001) 30:352-‐362.74 | P a g eGiving intravenous glutathione is actually a common practice and seems to help some patients. In others there is no response or they get worse. However, since no-one is cured by intravenous glutathione, it appears that it may be like beating a dying horse. He runs a little longer, and a little harder, but then eventually dies because the underlying problems have not been addressed. The safest way to increase glutathione is not by beating the horse, but by gently nurturing it back to life.So what’s the answer for Parkinson’s sufferers? Instead of trying to elevate glutathione by taking NAC or glutathione, the Parkinson’s patient should maintain a diet devoid of dietary free glutamates, and high in sulfur (cysteine is a sulfur- containing amino acid) and vitamin C. These two nutrients have been shown to increase glutathione dramatically.210,211The Need For Ascorbic AcidIn a 1975 study researchers tell of a 62-year old man who had done well on large doses of levodopa initially, and then intolerable side-effects caused him to terminate treatment. He was later restarted on a reduced level (3g) of levodopa along with ascorbic acid, 1 gram per day, and gradually increased to 4 grams, while at the same time, the L-dopa was reduced (2g) per day. It was reported that the patient “almost immediately” reported a decrease in side-effects. He was, in fact, able to return to doing the things he loved. To test that it was, indeed, the ascorbic acid causing the beneficial effects, the researchers put him on placebo and found that within two weeks the benefits disappeared. 212So how much vitamin C do we need? Dr. Irwin Stone produced a table showing how most mammals (except bats, humans, monkeys and guinea pigs) produce the ascorbic acid they need in their liver. On his table, it shows that a goat, which210 CS Johnston et al. Vitamin C elevates red blood cell glutathione in healthy adults. American Journal of Clinical Nutrition Vol 58 (1993) 103-‐105.
211 PG Paterson et al. Sulfur amino acid deficiency depresses brain glutathione concentration. Nutr Neurosci (2001) 4(3):213-‐222.212 William Sacks, George M. Simpson. Ascorbic Acid in Levodopa Therapy. The Lancet. March 1, 1975. P. 52775 | P a g ewould be about the size of a man, produces up to about 13 grams of vitamin C daily.213 Because of this, “Dr. Vitamin C”, Dr. Linus Pauling recommended between two and ten grams of vitamin C daily for people, at a time when most vitamin manufacturers were wary of producing supplemental vitamin C higher than about 100 mg. Mammals that do not produce their own vitamin C have lost one of the four enzymes (L-gulonolactone oxidase) that converts blood glucose into ascorbate in the liver.214Ascorbic acid has been shown to appreciably reduce the effect of MPTP neurotoxicity.215 Protein cross-linking seen in neurological diseases, was diminished in the presence of ascorbate.216It is known that ascorbic acid can behave as an antioxidant, but also as a pro- oxidant. The pro-oxidant ascorbic acid adds electrons to transition metals like copper and iron, which can generate superoxide and other reactive oxygen species. One study shows that intravenous use of vitamin C does not seem to increase pro- oxidant activity217 however in Parkinson’s patients the known elevation of iron may make intravenous vitamin C use unwise.Are Drugs an Insult to Injury?Long-term L-dopa therapy is known to lead to major complications. By now I think we can see that the evidence shows that dopamine isn’t causing the toxicity, but that toxicity is causing dopamine to oxidize. Without coming to that conclusion, however, researchers are searching for drug alternatives. One possibility is adenosine A2A receptor antagonists (A2A). A2A receptors in the213 I. Stone. New Dynamics of Preventive Medicine. 1974 2:19.
214 I. Stone. “Eight Decades of Scurvy. The Case History of a Misleading Dietary Hypothesis”. July 16, 1978.
215 H. Sershen et al. Protection against 1-‐Methyl-‐4-‐Phenyl-‐1,2,3,6-‐Tetrahydropyridine Neurotoxicity By The Antioxidant Ascorbic Acid. Neuropharmacology (1985) Vol 24. No. 12. 1257-‐1259.
216 Jennifer N. Rees et al. Protein Reactivity of 3,4-‐Dihydroxyphenylacetaldehyde, a Toxic Dopamine Metabolite, Is Dependent on Both the Aldehyde and the Catechol. Chem Res Toxicol (2009) 22:1256-‐ 1263.
217 A. Mühlhöfer et al. High dose intravenous vitamin C is not associated with an increase of pro-‐ oxidative biomarkers. European Journal of Clinical Nutrition (2004). 58(8):1151-‐1158.76 | P a g ebrain play important roles in the regulation of glutamate and dopamine release. A2A receptors are especially concentrated in brain areas naturally rich in dopamine. In pharmacology, when a receptor is upregulated within a disease, the attempt is to apply an antagonist as a therapeutic intervention. The problem is that the antagonist works in some cases, but not others, and worse, often causes serious, even life-threatening side-effects.Is there a better way than A2A antagonists? In the cascade of events stemming from mitochondrial damage, we see elevated levels of glutamate. As we’ve seen, glutamate is a neurotransmitter, which in excess is an excitotoxin, leading to neuronal death. Since studies have shown that A2A antagonists sometimes improves motor symptoms in Parkinson’s,218 it could very well be that it works by reducing the excitotoxic effects of glutamate. This is an important consideration, because glutamate excess is only one of many things going on in Parkinson’s. The better approach to controlling all of the things going on is not a drug that only partially works, and could be dangerous, but to stop “all the things going on” in the first place. Prevention and repair of the underlying mitochondrial damage again emerges as where our efforts should lie.To this day much confusion exists. Study after study will outline the pathological changes and motor dysfunctions seen in Parkinson’s; that oxidative stress is involved; that there are alterations in endogenous antioxidant systems like SOD and glutathione; that iron is present in excess; that lipids and proteins are oxidized; and that in the end, dopamine becomes toxic219 causing dopaminergic neurons to be killed. Researchers will examine each of these and wonder which of these events came first, and which is the “cause” of Parkinson’s. One study says that the “current major hypothesis is that nigral neuronal death in PD is due to excessive oxidative stress generated by auto and enzymatic oxidation of the endogenous neurotransmitter dopamine (DA).”220 Isn’t this just saying that Parkinson’s happens218 Micaela Morelli et al. Role of adenosine A2A receptors in parkinsonian motor impairment and L-‐DOPA-‐ induced motor complications. Progress in Neurobiology. (2007) 83:293-‐309.
219 Daniel Offen et al. Dopamine-‐melanin induces apoptosis in PC12 cells; possible implications for the etiology of Parkinson’s disease. Neurochem Int (Aug 3, 1997) 31(2):207-‐216.220 Ari Barzilai et al. Is There a Rationale for Neuroprotection Against Dopamine Toxicity in Parkinson’s Disease? Cellular and Molecular Neurobiology. (2001) Vol 21, No 3. 215-‐235.77 | P a g e
CAN WE REPAIR DAMAGED MITOCHONDRIA?”Mitochondrial membrane permeabilization” which involves the release of proteins from the mitochondrial intermembrane space, is commonly regarded as the “point of no return” in the cascade of events that lead to cell death.221 Unfortunately this would be why Parkinson’s and other neurological diseases involving mitochondrial damage are not much closer to being cured today than nearly 200 years ago at the time of James Parkinson’s essay. This whole mess, that we now know, begins with devastating damage to the mitochondria, has proven to be a “hard nut to crack.” But has this been because we’ve been barking up the wrong tree?One review discusses the “new role” of mitochondria in regulation of neuronal cell death of neurodegenerative disorders. They say that there are recent findings on new functions of mitochondria in regulation of their redox state and function through reversible S- glutathionylation. They go on to list the mitochondria as being at the beginning of the “apoptotic cascade”, which involves a defect of complex I detected in the outer membrane of the mitochondria which increases production of reactive oxygen and nitrogen species in mitochondrial oxidative phosphorylation system. Oxidative damage to proteins, lipids and DNA occur along with reduced anti-oxidant activities. The mitochondrial permeability transition is followed by release of apoptosis-inducing factors (AIFs) such as cytochrome c in cytoplasm, activation of caspase 9 and 3, and finally fragmentation and condensation of nuclear DNA. They say that the various genes in familial Parkinson’s (Parkin, PINK1, DJ-1, LRRK2, PARK8) directly or indirectly regulate mitochondrial function and integrity.222 Does mercury “mutate” the genes necessary for optimal functioning of the mitochondria in Parkinson’s and other “mitochondrial” diseases?Consider Blackinton et al, 2008, where they discuss DJ-1, a protein associated with “inherited Parkinsonism”. DJ-1 is normally protective of mitochondrial function in Parkinson’s. The researchers discuss how DJ-1 readily forms cysteine-sulfinic acid complex. They say that “Mutation of cysteine causes the protein to lose its normal protective function in cell culture and model organisms.” They go on to say, in effect, that they haven’t got a clue why the protein loses its protective function. They conclude that the formation of cysteine-sulfinic acid is a key modification221 Vladimir Gogvadze et al. Mitochondria as targets for chemotherapy. Apoptosis (2009) 14:624-‐640. 222 Makoto Naoi et al. Mitochondria in neurodegenerative disorders: regulation of the redox state and death signaling leading to neuronal death and survival. J Neural Transm September 18, 2009.79 | P a g ethat regulates the protective function of DJ-1. I propose that if we insert mercury into the equation as that which “mutates” the cysteine (molecular structure: C3H7NO2S, where “S” is sulfur not to mention the “sulfinic acid” it bonds with), we likely supply the missing puzzle piece.223Thankfully the research on mitochondrial dysfunction continues. In 2010, a study on human placental mitochondria, showed that the combination of melatonin with ascorbate with alpha-tocopherol nearly completely inhibited lipid peroxidation as seen in Parkinson’s.224CoQ10 is a potent free radical scavenger in the inner mitochondrial membranes.225 It participates in the production of ATP, and in reducing oxidative stress, thus blocking cell death.226 CoQ10 inhibits mitochondrial permeability transition pore known to cause cell death by increased retention of mitochondrial calcium.227 CoQ10 blocks nigrostriatal dopaminergic neurodegeneration in tests using various neurotoxic agents in various ages of lab mice.228A combined strategy of using creatine and CoQ10 has shown significant synergistic neuroprotective effects in laboratory studies.229 Creatine supplementation will increase already elevated nitric oxide in Parkinson’s, however, and should not be supplemented. Obtaining creatine from a diet containing high quality protein should probably be the only way to “supplement” creatine.223 Jeff Blackinton et al. Formation of a Stabilized Cysteine Sulfinic Acid is Critical for the Mitochondrial Function of the Parkinsonism Protein DJ-‐1. The Journal of Biological Chemistry. Vol. 284, No. 10. March 6, 2009.
224 Milczarek R et al. Melatonin enhances antioxidant action of alpha-‐tocopherol and ascorbate against NADPH-‐ and iron-‐dependent lipid peroxidation in human placental mitochondria.225 Beal MF. Bioenergetic approaches for neuroprotection in Parkinson’s disease. Ann Neurol 2003;53 (Suppl 3) S39-‐S47; discussion S47-‐S48.
226 Alleva R. et al. Coenzyme Q blocks biochemical but not receptor-‐mediated apoptosis by increasing mitochondrial antioxidant protection. FEBS Lett 2001;503:46-‐50.227 Papucci L et al. Coenzyme Q10 prevents apoptosis by inhibiting mitochondrial depolarization independently of its free radical scavenging property. J Biol Chem 2003;278:28220-‐28228.
228 Beal MF et al. Coenzyme Q10 attenuates the 1-‐methyl-‐4-‐phenyl-‐1,2,3,tetrahydropyridine (MPTP) induced loss of striatal dopamine and dopaminergic axons in aged mice. Brain Res 1998;783:109-‐114. 229 Yang et al., unpublished results as of 2010, as reported by Bobby Thomas PhD and M. Flint Beal, MD.80 | P a g eSome other mitochondrially-targeted therapies include MitoQ, currently being studied in New Zealand. MitoQ is the mitochondrially-targeted peptide antioxidant SS-31. It is a derivative of mitochondrial quinoline. It converts H2O2 to H2O and O2 to reduce toxicity from free radicals in the mitochondria. It has been shown to be effective in many in vivo models of mitochondrial dysfunction.230 Much more on mitochondrially-targeted antioxidants later.Some potential drug therapies being currently explored are PPAR-gamma coactivator 1alpha (PGC-1a) and the sirtuin family of enzymes both of which have been shown effective in mitochondrial biogenesis.231,232Mitochondrial TransplantResearchers at Newcastle University took the nucleus from a human embryonic cell and transplanted it into an anucleated human cell. This “swapped” the mitochondria, and other organelles in the cytoplasm. In essence, this would be similar to an organ transplant from another person to save a life. A reporter, writing about this, asked, “why not?” If someone with muscular dystrophy, multiple sclerosis, heart or any other “mitochondrial disorder” needed it to save their life, why wouldn’t you do a mitochondrial transplant? Might this be the answer for Parkinson’s?233 Apparently, the U.S. and Australia call this “cloning”. But the Council of Europe’s ‘Convention on Human Rights with Regard to Biomedicine’ states:Article 13 – Interventions on the human genome: An intervention seeking to modify the human genome may only be undertaken for preventive, diagnostic or therapeutic purposes and only if its aim is not to introduce any modification in the genome of any descendants.Well, this applies to Parkinson’s patients. Of course with what we know/suspect about mercury’s damage to the mitochondria, every Parkinson’s patient has a right to have a mitochondrial transplant to overcome the devastation caused to their230 Cocheme HM et al. Mitochondrial targeting of quinones: therapeutic implications. Mitochondrion 2007;7 (suppl):S94-‐S102.
231 McGill et al. PGC-‐1a, a new therapeutic target in Huntington’s disease? Cell 2006;127:465-‐468. 232 Nemoto S et al. SIRT1 functionally interacts with the metabolic regulator and transcriptional coactivator PGC-‐1(a). J Biol Chem 2005;280:16456-‐16460.233 Mitochondrial transplant for human embryos. Thursday, 14 February, 2008. http://hplusbiopolitics.wordpress.com/2008/02/14/mitochondrial-‐transplant-‐for-‐human-‐embryos81 | P a g ebody by mercury. Especially since they are damaged, due to no fault of their own, but to the fault of the government for allowing mercury in industry, dentistry, and medicine, long past when it’s devastating effects have been known and ignored.In 2010 the University of Newcastle’s Mitochondrial Research Group in England stated that treatment options for “mitochondrial myopathies” are limited, in spite of our current understanding of the problem! They do, however, state that beneficial to improving mitochondrial function are exercise training and CoQ10 above all. They say that there are other strategies, including the “ketogenic diet” that need more research.234 Of course, if we keep poisoning ourselves with mercury, all the benefits from any therapy will be for naught.Stem Cell TherapyAs of 2010 stem cell research continues. Results are encouraging, but not long- lasting. However, mitochondrial-dysfunction is not yet the primary focus.One study says: “They suggested two processes that put dopamine neurons at risk: the function of cellular machinery for degrading proteins and organelles, and oxidative stress secondary to uptake and sequestration of dopamine.”235 With all we’ve discussed heretofore, inserting mercury and mitochondrial dysfunction into these researchers’ equation would likely clear up a lot of the mystery.”Over the past quarter century, many experimental replacement therapies have been tried on PD animal models as well as human patients, yet none resulted in satisfactory outcomes… Eventually, nearly all of the dopaminergic neurons die of degeneration, and patients become unresponsive to L-dopa.”236 Even the title of this study tells the frustrating story: “Cells therapy for Parkinson’s disease – so close and so far away.”And why “so far away”, indeed? Without addressing dysfunctional mitochondria that are spewing out deadly free radicals, and repairing components of mitochondrial respiration that are not functioning properly, and somehow replacing normally protective endogenous antioxidants that are damaged, no amount of stem234 Hassani A et al. Mitochondrial myopathies: developments in treatment. Curr Opin Neurol 2010 Jul 21. 235 Collier TJ et al. Presidential symposium: aging and Parkinson’s disease: the connection revisited. Cell Transplant 2008;17:457.
236 Ren ZhenHua & Zhang Yu. Cells therapy for Parkinson’s disease -‐ so close and so far away. Sci China Ser C-‐Life Sci, 2009. 52(7): 610-‐614.82 | P a g ecell therapy will likely ever work! Both supplying more and more L-dopa, only to have it oxidize and become part of the problem and transplanting stem cells that may thrive for a while, but then die as well, is like seeing someone shooting at fish in a fish barrel, the fish dying and their guts spattering everywhere. Is the answer to stock the barrel with more fish? Or stop the shooter?The focus of stem cell therapy must be on repairing/replacing mitochondria or 100 years from now we will still be scratching our heads. Of course, prior to stem cell therapy, the focus must be on the complete elimination of mercury’s use in medicine, dentistry and industry (or at least insomuch as it is then spewed into the environment).Nutritional Supplements For Parkinson’sUntil such time as a Parkinson’s patient can walk into a doctor’s office and order a mitochondrial transplant, powerful preventative and even healing therapy can come from the right supplements, foods & therapies. We start here with supplements. Many of the supplements are “polyphenols” discussed in more detail in the next section. By concentrating food-source polyphenols, a more therapeutic dose can be achieved. Neurological diseases come about by extraordinary means as discussed, and extraordinary measures must be taken to counteract the damage.Phytochemicals, especially polyphenols have been studied by thousands of researchers. Researchers have found that polyphenols can behave as pro-oxidants sometimes, and anti-oxidants at other times. For example, polyphenols have been found to generate reactive oxygen species (like nitric oxide), especially in the presence of transition metals (like iron), and shown to arrest the proliferation of tumor cells.237,238 Then at other times polyphenols function either directly or indirectly as antioxidants.239,240,241 Because the brain is low in natural antioxidant properties, and high in natural transition metals (iron and copper), supplements237 CA de la Lastra, I Villegas. Resveratrol as an antioxidant and pro-‐oxidant agent: mechanisms and clinical implications. Biochem Soc Trans (2007) 35:1156-‐1160.
238 KW Lee, HJ Lee. The roles of polyphenols in cancer chemoprevention. Biofactors (2006) 26:105-‐121. 239 B Frei, JV Higdon. Antioxidant activity of tea polyphenols in vivo: evidence from animal studies. J Nutr (2003) 133:3275S-‐3284S.
240 KE Heim et al. Flavonoid antioxidants: chemistry, metabolism and structure-‐activity relationships. J Nutr Biochem (2002) 13:572-‐584.
241 T. Nakagawa, T Yokozawa. Direct scavenging of nitric oxide and superoxide by green tea. Food Chem Toxicol (2002) 40:1745-‐1750.83 | P a g ehave to be chosen carefully. Taking a high concentration of a single supplement can prove counterproductive at best, and harmful at worst.A supplement that might appear helpful for one of the things going on might horribly exacerbate another of the things going on. In neurological diseases, within the cascade of events stemming from a damaged mitochondria, there are elevated levels of nitric oxide. For example, a supplement recommended for Parkinson’s because it stimulates dopamine production (just to use as an example) might also stimulate nitric oxide production, and could therefore be detrimental. In addition, going to extreme efforts to supply every amino acid or protein compound known to repair cells can actually exacerbate symptoms and lead to further suffering, pain, and damage.Also, when choosing a supplement to combat Parkinson’s, does it even get to the intended target – the brain? Studies following polyphenols within the body find that metabolism of polyphenols involves absorption in the small intestine, and processing in the liver to generate active forms that reach the tissues in a form that can be utilized (glucoronidated, methylated, and sulfated, for example).242 This is what is meant by a substance being “bioavailable”. Once the polyphenol is bioavailable we run into the issues of uptake by tissues and cells, which varies from polyphenol to polyphenol. Then comes the issue of whether or not that bioavailable polyphenol can cross the blood-brain barrier. And, perhaps as a gift of God and Nature, it has been shown that most polyphenolic compounds are able to permeate the blood-brain barrier and therefore do their antioxidant magic directly in the brain.243,244,245,246Hormetic Pathway – Vitagenes
Some supplements have powerful healing and neuroprotective effects by activationof hormetic pathways. “Hormesis” means to “set in motion”, even impel or urge242 C Manach et al. Polyphenols: food sources and bioavailability. Am J Clin Nutr (2004) 79:727-‐747.
243 S. Mandel et al. Green tea catechins as brain-‐permeable, natural iron chelators-‐antioxidants for the treatment of neurodegenerative disorders. Mol Nutr Food Res (2006) 50:229-‐234.
244 KA Youdim et al. Flavonoids and the brain: interactions at the blood-‐brain barrier and their physiological effects on the central nervous system. Free Radic Biol Med (2004) 37:1683-‐1693.
245 M Mokni et al. Effect of resveratrol on antioxidant enzyme activities in the brain of healthy rat. Neurochem Res (2007) 32:981-‐987.
246 MM Abd El Mohsen et al. Uptake and metabolism of epicatechin and its access to the brain after oral ingestion. Free Radic Biol Med (2002) 33:1693-‐1702.84 | P a g eon. Used in physiology, a hormetic pathway is meant to mean that a biological response has been set in motion against exposure to a toxin or other stressor. In hormesis, the thought is that a response to low dose toxic insult is generally favorable, and the response to a high dose toxic insult would generally be unfavorable – even deadly.247 Homeopathic medicine is based upon the concept of the body being stimulated into repair mode in response to a low dose toxin.Of course survival against toxins would depend upon systems within the body capable of protection and repair. It is a complex system. The part of hormesis that involves longevity within the body’s protection and repair system, consists of several genes which have been termed “vitagenes”. When the vitagenes network begins to fail, it morphs into a “gerontogene” network, which puts the body on the pathway to aging.248 Our hope is that with a powerful antioxidant diet, supplements (including herbs) and lifestyle, we can reverse the hormetic pathway of gerontogene, back to one of vitagene. This is especially important in our discussion here on the use of dietary antioxidants in the treatment of neurodegenerative disorders like Parkinson’s. Recent studies show that there are many phytochemicals that have been found to be protective through the activation of the vitagenes hormetic pathway.BioavailabilityUltimately, finding the purest and most bioavailable form of needed supplements will be the crucial factor. Consider that when scientists study nutrients in the lab setting, the nutrient is often injected right near the cell to be tested and the results then watched and commented upon. Whereas in the body, some substances injected or taken orally might not reach the cells needing to be effected. One reason is that the blood-brain barrier keeps many substances out of the central nervous system (brain, spinal cord and retina). By the way this is not true of most “phytonutrients”, or plant nutrients (directly from the plant foods, eaten in their natural state), as they are found to cross the blood-brain barrier.In Parkinson’s, it is important to know which supplements are antagonistic to (oppose) nitric oxide and glutamate. We want to use supplements that are agonistic247 Calabrese EJ, Baldwin LA. The frequency of U-‐shaped dose responses in the toxicological literature. Toxicol Sci (Aug 2001) 62(2):330-‐338.
248 Sureshi I.S. Rattan. The Nature of Gerontogenes and Vitagenes: Anti-‐aging Effects of Repeat Heat Shock on Human Fibroblasts. Annal of the New York Academy of Sciences. Vol. 854. Towards Prolongation of the Healthy Life Span: Practical Approaches to Intervention 54-‐60. Nov. 1998.85 | P a g eto (encourage) endogenous (manufactured within the body) dopamine (both the production of dopamine and protection of dopamine from oxidation) and increase endogenous antioxidants like SOD and glutathione without increasing glutamate activity or nitric oxide. Above all, we want supplements and foods that support that which heals the mitochondria and the blood-brain barrier.The goal is not a lot of supplements, but the right supplements. We list most of the supplements known to be helpful in Parkinson’s. Reading through each, you and your physician can determine which should be a part of your regimen based upon things like have they been shown to cross the blood-brain barrier, can you get the substance in foods instead, is the substance available as a supplements, and of course, should you avoid them because they increase nitric oxide or glutamate.Acetyl-L-Carnitine an acetylated form of L-carnitine, also called ALCAR Blood-Brain Barrier acetyl-L-carnitine does cross the blood-brain barrier.Typical Dose During exercise, portions of L-carnitine and acetyl-coenzyme A are converted to acetyl-L-carnitine inside the mitochondria by carnitine O- acetyltransferase.249 L-Carnitine/acetyl-L-carnitine have been demonstrated in the lab to be neuroprotective through the activation of hormetic pathways, including vitagenes.250Alpha-Lipoic Acid – a thiol also called “thioctic acid”, abbreviated ALA. This supplement is widely available in capsule form.Made up of 2 sulfur atoms and a perhydroxyl radical (HO2)Blood-Brain Barrier Alpha-Lipoic Acid does cross the blood-brain barrier in a dosage and frequency dependent manner.251Typical Dose 200-400 mg./day249 Zeyner A, Harmeyer J. Metabolic functions of L-‐carnitine and its effects as feed additive in horses. A review. Archiv Fur Tierenahrung 52(2):115-‐138.
250 Vittorio Calabrese et al. Vitagenes, dietary antioxidants and neuroprotection in neurodegenerative diseases. Frontiers in Bioscience 14, 376-‐397, January 1, 2009. 376-‐397.251 Rooney, James. The role of thiols, dithiols, nutritional factors and interacting ligands in the toxicology of mercury. Toxicology 234(3):145-‐156.86 | P a g eAppears to help strengthen a weak blood-brain barrier.252 Converts to various forms within the body, one of which is an essential cofactor of four mitochondrial enzyme complexes.253 Reduces tissue levels of nitric oxide and increases glutathione.254 Also assists in restoring superoxide dismutase, catalase and myeloperoxidase activity.255 Treatment with lipoic acid prevents oxidative stress from nitric oxide produced by submitochondrial membranes damaged by lipopolysaccharide in experimental animals.256 Because of the sulfur, ALA appears to be a mercury chelator.257ALA may be more effective than glutathione at removing mercury from the brain, the body doesn’t produce nearly as much of it as it does glutathione.Alpha Tocopherol – also known as vitamin E and a-tocopherol. Blood-Brain Barrier
Typical DoseAlpha-tocopherol inhibits apoptosis induced by L-glutamine.258 Alpha-tocopherol along with ubiquinol are found in the mitochondria, and are particularly effective in scavenging lipid peroxyl radicals as well as preventing the free radical chain reaction of lipid peroxidation.259252 Schreibelt G et al. Lipoic acid affects cellular migration into the central nervous system and stabilizes blood-‐brain barrier integrity. J. Immunol. 177(4):2630-‐2637. August 2006.
253 Biewenga G Ph et al. An overview of Lipoate Chemistry, Chapter 1: Lipoic Acid in Health & Disease. 254 Emmez H et al. Anti-‐apoptotic and neuroprotective effects of alpha-‐lipoic acid on spinal cord ischemia-‐reperfusion injury in rabbits. Acta Neurochir (Wien). June 10, 2010.255 Saad El et al. Role of oxidative stress and nitric oxide in the protective effects of alpha-‐lipoic acid and amino guanidine against isoniazid-‐rifampicin-‐induced hepatotoxicity in rats. Food Chem Toxicol. April 22, 2010.
256 Vanasco V et al. The oxidative stress and the mitochondrial dysfunction caused by endotoxemia are prevented by alpha-‐lipoic acid. Free Radic Res 2008, Sep;42(9):815-‐823.257 Gregus, Z et al. Effects of alpha-‐lipoic acid supplementation on biliary excretion of glutathione and metals. Toxicology and Applied Pharmacology 114(1):88-‐96.
258 Svoboda N, Kerschbaum HH. L-‐Glutamine-‐induced apoptosis in microglia is mediated by mitochondrial dysfunction. Eur J Neurosci 2009 Jul;30(2):1960206.259 Hazel H. Szeto. Mitochondria-‐Targeted Peptide Antioxidants: Novel Neuroprotective Agents. The AAPS Journal 2006;8(3) Article 62.87 | P a g eDaily oral supplementation of a-tocopherol in rats prevented impairment in maze performance when they were infused with amyloid-beta (a peptide of 39–43 amino acids that appears to be the main constituent of amyloid plaques in the brains of Alzheimer’s disease patients, and Parkinson’s disease dementia).260Ascorbic Acid also known as vitamin C This vitamin is widely available in supplement form.Blood-Brain Barrier Typical DoseAstaxanthinBlack Tea ExtractsBlood-Brain BarrierTypical DoseBlack tea extract constituents acted as dietary chelators with the potential to form complexes mimicking the endogenous antioxidant super oxide dismutase activities.261CaffeineBlood-Brain BarrierTypical DosePretreatment with caffeine against 6-OHDA-lesioned rats showed that caffeine attenuated apopmorphine-induced rotational behavior. Apomorphine is used to260 K Yamada et al. Protective effects of idebenone and alpha-‐tocopherol on beta-‐amyloid (1-‐42)-‐induced learning and memory deficits in rats: implication of oxidative stress in beta-‐amyloid-‐induced neurotoxicity in vivo. Eur J Neurosci (1999) 11:83-‐90.
261 RK Chaturvedi et al. Neuroprotective and neurorescue effect of black tea extract in 6-‐ hydroxydopamine-‐lesioned rat model of Parkinson’s disease. Neurobiol Dis (2006) 22:421-‐434.88 | P a g etreat “off” episodes (times of difficulty moving, walking, and speaking that occurs in Patient’s with Parkinson’s, when their L-dopa wears off.262CarnosineBlood-Brain BarrierTypical DoseCarnosine has been shown in lab studies to be neuroprotective through the activation of the vitagenes hormetic pathway.263Coenzyme Q10 also known as CoQ10, ubiquinone or its more reduced and active form ubiquinol (a process normally done within the mitochondria). This supplement is widely available in supplement form.An oil-soluble substance made within the body, and present in foods as well.Blood-Brain Barrier Properly emulsified and micellized CoQ10 crosses the blood- brain barrier. Damage to the mitochondria increases requirements for CoQ10 and supplementation of forms that will readily cross the blood-brain barrier and mitochondrial membrane.Typical Dose 150 mg Ubiquinol equals 1200 mg Ubiquinone264 It was 1200 mg of Ubiquinone that was found in a 2002 study to reduce the progression of Parkinson’s by 44%.265Coenzyme Q10 is found in small amounts in foods, but due to mitochondrial damage , there is a need for therapeutic doses, and so supplementation is necessary. Coenzyme Q10 is present in most cells, primarily in the mitochondria. It is part of the electron transport chain across the membrane of the mitochondria, participating in cellular respiration, and generating energy in the262 MT Joghataie et al. Protective effect of caffeine against neurodegeneration in a model of Parkinson’s disease in rat: behavioral and histochemical evidence. Parkinsonism Relat D (2004) 10:465-‐468.
263 Vittorio Calabrese et al. Vitagenes, dietary antioxidants and neuroprotection in neurodegenerative diseases. Frontiers in Bioscience 14, 376-‐397, January 1, 2009. 376-‐397.264 Yan J, Fuji K et al Reduced coenzyme Q10 supplementation decelerates senescence in SAMP1 mice. Exp Gerontol. 2006 Feb;41(2):130-‐140.
265 Clifford W. Shults MD et al. Effects of coenzyme Q10 in early Parkinson disease: evidence of slowing of the functional decline. Archives of Neurology, 59, No. 10, (October 2002) 1541-‐155089 | P a g eform of ATP. Ninety-five percent of the human body’s energy is generated in this way.266 There are various kinds of Coenzyme Q, but it is Coenzyme Q10 that is most prevalent in the mitochondria. The 10 refers to a molecular unit that makes up the molecule and is repeated 10 times. Emulsification and micelle formation is required for CoQ10’s absorption. Micelles are lipid molecules that arrange themselves in a spherical form in aqueous solutions. This process is mostly done by secretions from the pancreas and bile salts in the small intestine. Due to poor absorption across the blood-brain barrier, researchers are rapidly working on a form that will enter the mitochondria. One formulation that completed its clinical trials July 2010, is MitoQ produced by Antipodean Pharmaceuticals in New Zealand. MitoQ is CoQ10 covalently bonded to triphenylphosphonium (TPP+) cation.267Creatine increases nitric oxide.Creatine is a guanidine compound, normally found in urine as a byproduct of protein metabolism. Creatine is a reservoir for ATP. It is possible that creatine enables mitochondrial creatine kinase to remain in a stable (octameric) condition to where mitochondrial permeability transition pore is inhibited, thus blocking cell death.268 If mitochondrial creatine kinase converts to a less stable (dimeric) state, the permeability transition pore opens leading to cell death.269 But if this is not the way creatine prevents cell death, another possibility is that both creatine and phosphocreatine enhance cytosolic high energy phosphates that maintain ATP levels during oxidative stress-induced neurodegeneration.270,271 Creatine is made in the body from other amino acids in the liver, kidney and pancreas. The rest is266 Dutton PL, et al 4 Coenzyme Q oxidation reduction reactions in mitochondrial electron transport (pp 65-‐82) in Coenzyme Q: Molecular mechanisms in health and disease edited by Kagan VE and Quinn PJ, CRC Press (2000), Boca Raton
267 Andrew M. James et al. Interaction of the Mitochondria-‐targeted Antioxidant MitoQ with Phospholipid Bilayers and Ubiquinone Oxiodoreductases. The Journal of Biological Chemistry. Dec 14,2006.
268 Dolder M et al. Inhibition of the mitochondrial permeability transition by creatine kinase substrates. Requirement for microcompartmentation. J Biol Chem 2003;278:17760-‐17766.
269 O-‐Gorman E et al. The role of creatine kinase in inhibition of mitochondrial permeability transition. FEBS Lett 1997;414:253-‐257.
270 Klivenyi P et al. Neuroprotective mechanisms of creatine occur in the absence of mitochondrial creatine kinase. Neurobiol Dis 2004;15:610-‐617.
271 Brustovetsky N et al. On the mechanisms of neuroprotection by creatine and phosphocreatine. J Neurochem 2001;76:425-‐434.90 | P a g eobtained from protein foods like poultry and lean red meat. Creatine should absolutely not be supplemented where there is kidney dysfunction. Excess creatine can cause the body to lower or stop producing its own.272Curcumin Botanical name: Curcuma longa L. Chemical name: diferuloylmethane. A polyphenol found in the root or stem of the herb Turmeric. This herb is widely available in supplement form.Blood-Brain BarrierTypical Dose One study used 80 mg/kg as a therapeutic dose for 3 days (About 4 grams in a 110 pound person)In the Indian culture, where curcumin is used widely in foods and medicines, there is a 4.4-fold reduced prevalence in Alzheimer’s disease compared to the United States.273 Curcumin scavenges superoxide anions and nitric oxide radicals274 seen in Parkinson’s, and inhibits lipid peroxidation275. Curcumin protects the mitochondria from peroxynitrite damage, restores depleted glutathione and preserves mitochondrial Complex I activity.276While the toxin paraquat enhances cell death via the production of reactive oxygen species and thus oxidative stress damage to cells277, curcumin inhibits reactive oxygen species, thus protecting against cell death by paraquat.278Curcumin has been shown in lab studies to be neuroprotective through the activation of the vitagenes hormetic pathway.279272 University of Maryland Medical Center. Creatine. http://www.umm.edu/altmed/articles/creatine-‐ 000297.htm
273 M. Ganguli et al. Apolipoprotein E polymorphism and Alzheimer’s disease: the Indo-‐US Cross-‐National Dementia Study. Arch Neurol (2000) 57:824-‐830.
274 Sreejayan N, Rao MN. Nitric oxide scavenging by curcuminoids. Pharm Pharmacol 1997;49:105-‐7. 275 Sreejayan N, Rao MN. Free radical scavenging activity of curcuminoids. Arzneimittelforschung 1996; 46:169-‐171.
276 Balusamy Jagatha et al. Curcumin treatment alleviates the effects of glutathione depletion in vitro and in vivo: Therapeutic implications for Parkinson’s disease explained via in silico studies. Free Radical Biology and Medicine (March 2008) Vol 44, Issue 5. 907-‐917.
277 Thrash B et al. Paraquat and maneb induced neurotoxicity. Proc West Pharmacol Soc 2007;50:31-‐42. 278 J. Chen et al. Curcumin protects PC12 cells against 1-‐methyl-‐4-‐phenylpyridinium ion-‐induced apoptosis by bcl-‐2-‐mitochondria-‐ROS-‐iNOS pathway. Apoptosis. 2006;11:943-‐953.91 | P a g eCurcumin reduced amyloid-beta deposits and prevented spatial memory deficits induced by infusions of amyloid-beta.280Curcumin is a dietary antioxidant chelator which has shown SOD-like activity.281 Curcumin has a protective effect with routine dietary intake against mercuryexposure.282
Ganoderma Lucidum Reishi Mushroom, Lingzhi This mushroom is widelyavailable in supplement form.Blood-Brain Barrier Typical DoseThis mushroom has been used in traditional Chinese medicine for more than 4,000 years. It has been shown to have anti-bacterial, anti-viral and anti-fungal properties. 283,284Microglia were isolated and purified from brains of infant Wistar rats. The microglia were activated by lipopolysaccharide and MPP+ treated MES 23.5 cell membranes. In a dose-dependent manner, extract of Reishi mushroom significantly prevented microglia-derived proinflammatory and cytotoxic factors: Nitric oxide, tumor necrosis factor-a (TNFa), interlukin 1β (IL-1β). The extract down-regulated TNFa and IL-1β expression at the mRNA level as well.285279 Vittorio Calabrese et al. Vitagenes, dietary antioxidants and neuroprotection in neurodegenerative diseases. Frontiers in Bioscience 14, 376-‐397, January 1, 2009. 376-‐397.
280 SA Frautschy et al. Phenolic anti-‐inflammatory antioxidant reversal of Abeta-‐induced cognitive deficits and neuropathology. Neurobiol Aging (2001) 22:993-‐1005.281 A Bank et al. Evaluation of a new copper(II)-‐curcumin complex as superoxide dismutase mimic and its free radical reactions. Free Radical Biol Med (2005) 39:811-‐822.
282 R Agarwal et al. Detoxification and antioxidant effects of curcumin in rats experimentally exposed to mercury J Appl Toxicol (Jul 2010) 30(5):457-‐468.283 Wang H, Ng TB. Ganodermin, an antifungal protein from fruiting bodies of the medicinal mushroom Ganoderma lucidum. Peptides (January 2006) 27(1):27-‐30.
284 Moradali MF et al. Investigation of potential antibacterial properties of methanol extracts from fungus Ganoderma applanatum. Chemotherapy (2006) 52(5):241-‐244..285 Ruiping Zhang et al. Ganoderma lucidum Protects Dopaminergic Neuron Degeneration Through Inhibition of Microglial Activation. eCAM 2009 1-‐9. email@example.com | P a g eGinkgo Biloba also known as maidenhair tree. Can also be spelled gingko. This herb is widely available in supplement form.Blood-Brain Barrier Studies show that after the administration of ginkgo, its flavonoid metabolites (quercetin, kaempferol and isorhamnetin derivatives)increase 2-fold in the brain, with about 90% distributed in the hippocampus, frontal cortex, striatum and cerebellum (38% of the brain).286Typical Dose The studies have used 50 to 600 mg/kg body weight.Ginkgo trees are very large, reaching up to over 100 feet and are mostly grown in China. During the Fall, the tree leaves turn bright yellow, then fall. The trees are highly resistant to insects and disease, making them very long-lived, with some trees thought to be up to 2,500 years old!The female trees produce seeds from which the extract is obtained. This extract has been found to contain potent antioxidant properties as well as natural monoamine oxidase B (MAO-B) inhibitor. The terpene lactones and flavonoid glycosides of ginkgo biloba are credited for its pharmacological actions.The natural MAO-B inhibitor has been studied for anti-parkinsonian effects in a 6- hydroxydopamine rat model of Parkinson’s. After several weeks on Ginkgo, the animals’ drug-induced symptoms of Parkinson’s were significantly (and dose- dependently) reduced by Ginkgo. Ginkgo restored glutathione as well as the activities of glutathione-dependent enzymes catalase and SOD in the striatum. Ginkgo reversed the decrease in number of dopaminergic D2 receptors in the striatum.287Ginkgo has been shown to have direct protective effects against mitochondrial dysfunction in platelets. In the hippocampi, these protective effects were observed286 Rangel-‐Ordonez L et al. Plasma Levels and Distribution of Flavonoids in Rat Brain after Single and Repeated Doses of Standardized Ginkgo biloba Extract EGb 761(R). Planta Med 2010 May 19. Epub.
287 Muzamil Ahmad et al. Ginkgo biloba affords dose-‐dependent protection against 6-‐hydroxydopamine-‐ induced parkinsonism in rats: neurobehavioural, neurochemical and immunohistochemical evidences. Journal of Neurochemistry, 2005, 93, 94-‐104.93 | P a g eonly in the elderly. Researchers decided this may be due to an increase in blood- brain barrier permeability that comes with age.288Ginsenoside a ginseng saponin derivative from Ginseng. Readily available in Ginseng supplements.Blood-Brain Barrier Typical DoseGinsenosides are actually a family of 31 known “triterpene saponins”. Each ginsenoside has been given a label, like Rp1 or Rg1. Ginsenosides from both American and Asian Ginseng are widely used to manage acute and chronic inflammatory diseases in Asian cultures.Ginsenoside Rp1 has been found to exert its powerful antiinflammatory effect due to inhibition of interleukin-1β production by inhibiting the NF-ӄB pathway.289NF-ӄB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a protein complex that controls the transfer of DNA sequences to mRNA. NF-κB is found in almost all animal cell types and is also involved in the cell’s response to negative stimuli. As such, NF-ӄB is seen to jump into action during the cascade of events leading to inflammation both acute and chronic. NF-ӄB is also involved in the synapsing of neurons.L-Theanine a non-sulfur amino acid. Also called 5-N-ethyl glutamine. It differs from glutamine by a CH2-CH3 (ethyl) group. This amino acid is best obtained from sweet green tea, but is also widely available in supplement form.Blood-Brain Barrier L-Theanine does cross the blood-brain barrier.290Typical Dose 20-300 mg/day. It is thought that a cup of tea containing theanine likely only contains about 5 mg. per cup, but this is difficult to quantify because288 Shi C et al. Ginkgo biloba extract EGb761 protects against aging-‐associated mitochondrial dysfunction in platelets and hippocampi of SAMP8 mice. Platelets. 2010;21(5):373-‐379.
289 Byung Hun Kim et al. Ginsenoside Rp1, a Ginsenoside Derivative, Blocks Lipopolysaccharide-‐Induced Interleukin-‐1β Production via Suppression of the NF-‐ӄB Pathway. Planta Med 2009; 75:321-‐326.290 Yokogoshi H et al. Effect of theanine, r-‐glutamylethylamideon brain monoamines and striatal dopamine release in conscious rats. Neurochem Res 23(5):667-‐673.94 | P a g edifferent green teas have different levels, and the person brewing the tea may steep it varying lengths of time. Gyokuro is a “sweet green tea” that is supposed to be the highest quality green tea, and highest in therapeutic compounds.L-Theanine is either extracted from green tea, or synthesized the same way as in tea leaves. To synthesize, an ATP added to ethylamine and glutamate yields ADP, a phosphate and theanine. “Suntheanine” is touted as the purest synthesized L- Theanine. L-Theanine protects neurons from excess glutamate by blocking glutamate’s entrance into cells, thus it inhibits excitotoxicity seen in neurological diseases. Decreases nitric oxide production resulting from down-regulated protein levels of iNOS and nNOS, indicating the inhibition of the NMDA subtype of glutamate receptors which would account in part for the neuroprotective effect of L-theanine.291 Attenuates the down-regulation of BDNF and GDNF production in SH-S&5& cells suggesting a direct neuroprotective action against Parkinson’s disease-related neurotoxicants.292 This is thought to be done, at least in part, by increasing levels of GABA (gamma-amino-butyric acid) an important inhibitory neurotransmitter.293 Theanine increases brain dopamine levels as well as has an affinity for AMPA, kainate and NMDA receptors.294Caution! It would likely be best for someone with mitochondrial dysfunction, to get their L-Theanine in its natural form, from high quality sweet green tea. Isolating L-Theanine, especially taken in the high dose found in supplements, will likely prove too disruptive to the “good guy” “bad guy” activity of nitric oxide, lowering the amount needed for healthy cardiovascular function. As with so many things in Nature, obtaining nutrients in their natural state is best because there are other compounds designed by Nature to work synergistically with the one nutrient. L-Theanine, in its natural state is surrounded by polyphenols (flavonols),291 X Di et al. L-‐Theanine Protects the APP (Swedish Mutation) Transgenic SH-‐SY5Y Cell Against Glutamate-‐Induced Excitotoxicity via Inhibition of the NMDA Receptor Pathway. Neuroscience (2010). 292 Cho HS et al. Protective effect of the green tea component, L-‐theanine on environmental toxins-‐ induced neuronal cell death.
293 Subhuti Dharmananda PhD, Director for Traditional Medicine, Portland, Oregon. Amino Acid Supplements: Theanine.
294 Nathan P et al. The neuropharmacology of L-‐theanine (N-‐ethyl-‐L-‐glutamine): a possible neuroprotective and cognitive enhancing agent. J Herb Pharmacother 6(2):21-‐30.95 | P a g ecommonly known as the catechins EGCG, EGC, ECG, EC plus caffeine (an alkaloid) to name a few synergistic compounds.295NADRetinoids And Carotenoids There are two types of vitamin A, retinoids and carotenoids. These nutrients are best obtained from the diet, but beta-carotene and vitamin A are widely available in supplement form.Blood-Brain Barrier Typical DoseRetinoids are the fat-soluble form of vitamin A and are found in animal-source foods like liver, fish oils and butter. Or retinoids can be made within the body just as they are made in the body of animals that are consumed. Carotenoids are found in plants like carrots and broccoli, and other dark green, orange, yellow or red vegetables. Carotenoids also convert to vitamin A in the body, but only as much as is needed.Studies have shown that retinoids bind to retinoic acid receptors (RAR) and peroxisome proliferator-activated receptor (PPAR). RAR and PPAR are ligand- activated transcription factors which have been recently implicated in the progression of neurodegenerative and psychiatric diseases. Studies with vitamin A deprivation have given scientists evidence that retinoic acid signaling is directly involved in the action and health of motoneurons. 296Rhodiola Rosea an herb also called Golden Root, Roseroot, Aaron’s Rod. This is an herb that is widely available in supplement form.Blood-Brain Barrier Typical DoseDepressive rats (induced by chronic mild stress) were given 1.5 grams/kg of Rhodiola rosea extract for three weeks. Their serotonin (5-HT) levels recovered to295 Chung S. Yang, Zhi-‐Yuan Wang. The Chemistry of Green Tea. http://www.teatalk.com/science/chemistry.htm
296 Savien van Neerven et al. RAR/RXR and PPAR/RXR signaling in neurological and psychiatric diseases. Progress in Neurobiology 85 (2008) 433-‐451.96 | P a g enormal status. Researchers concluded that Rhodiola rosea can induce neural stem cell proliferation at hippocampus to return to normal level, and repair injured neurons at hippocampus.297SeleniumBlood-Brain Barrier Typical DoseSelenium was protective against 6-OHDA-induced Parkinson’s.298 When selenium was given with or without vitamin E it reduced methyl mercury toxicity.299,300 When selenium as selenite (gypsum) was co-administered, there was a four-fold protection against methyl mercury over selenate (the sodium salt of selenic acid).301 It has been found that the protective effect of mercury occurs prior to absorption.302 Selenium isn’t something to be aggressively supplemented, but is better obtained from the right diet. When administered in conjunction with inorganic mercury (Hg2+), selenium protects against mercury toxicity. When administered alone selenium can methylate to the toxic dimethylselenide.282 Selenium is more efficiently absorbed as selenate than as selenite. (94% versus 59%)303 Selenium in nature doesn’t exist alone. Supplements of selenium contain forms more like those found in nature, most often L-selenomethionine (selenium + the amino acid methionine). Dietary consumption of selenium reduced the amount of mercury absorbed from mercury-contaminated fish by 5-11%, but had no effect on methyl mercury absorption from water.286297 Q.G. Chen et al. The effects of Rhodiola rosea extract on 5-‐HT level, cell proliferation and quantity of neurons at cerebral hippocampus of depressive rats. Phytomedicine 16 (2009) 830-‐838.
298 KS Zafar et al. Dose-‐dependent protective effect of selenium in rat model of Parkinson’s disease: neurobehavioral and neurochemical evidences. J Neurochem (2003) 84:438-‐446299 J Gailer et al. Structural basis of the antagonism between inorganic mercury and selenium in mammals. Chemical Research in Toxicology, 13, (2000) 1135-‐1142.
300 P Beyrouty & HM Chan. Co-‐consumption of selenium and vitamin E altered the reproductive and developmental toxicity of methylmercury in rats. Neurotoxicol Teratol (2006)
301 M Kasuya. Effect of selenium on the toxicity of methylmercury on nervous tissue in culture. Toxicol Appl Pharmacol, 35, (1976)11-‐20.
302 ML Cuvin-‐Aralar & RW Furness. Mercury and selenium interaction: a review. Ecotoxicol Environ Saf, 21, (1991)348-‐364.
303 CD Thomson & MF Robinson Urinary and fecal excretions and absorption of a large supplement of selenium: Superiority of selenate over selenite. Am. J.Clin. Nutr. 44, (1986) 659-‐663.97 | P a g eSilymarin Silibinin (also known as silybin) is the major active component in silymarin, a mixture of flavonolignans* extracted from an herb commonly knownas Blessed Milk Thistle (botanical name Silybum marianum). Lesser known components of the herb include Silibinin B, isosibilinin A and B, Silicristin and Silidianin. [*Flavonolignans are polyphenols composed of part flavonoid and part lignan. Lignans are phytochemicals known as phytoestrogens, which are estrogen- like chemicals which also act as antioxidants.]Blood-Brain Barrier Typical DoseSilymarin significantly inhibits the LPS-induced activation of microglia (mediated through the inhibition of nuclear factor kappaB activation) and resultant production of inflammatory mediators, such as tumor necrosis factor alpha (TNFa) and nitric oxide. Silymarin effectively reduced LPS-induced superoxide generation and nuclear factor kappaB (NF-kappaB) activation. This reduced damage to dopaminergic neurons.304Tripterygium Wilfordii an herb, also known as Thunder God Vine, is used in Chinese medicine for everything from fever to carbuncles. Not generally available in supplement form.Blood-Brain Barrier Typical DoseTriptolide is the active component in the herb Tripterygium Wilfordii that has been shown to inhibit NF-ӄB transcriptional activity, and thus induce apoptosis in tumor cells. Triptolide has immunosuppressive, antiinflammatory and antitumor activity. Because triptolide suppressed the activation of microglia significantly, it protected dopaminergic neurons from inflammation-mediated damage.305,306 At medicinal levels of administration, side effects have been lowered production of304 Wang MJ et al. Silymarin protects dopaminergic neurons against lipopolysaccharide-‐induced neurotoxicity by inhibiting microglia activation. Eur J Neurosci 2002 Dec; 16(11):2103-‐2112.
305 Hui-‐Fang Zhou et al. Triptolide protects dopaminergic neurons from inflammation-‐mediated damage induced by lipopolysaccharide intranigral injection. Neurobiology of Disease 18 (2005) 441-‐449.
306 Gao JP et al. Triptolide protects against 1-‐methyl-‐4-‐phenyl pyridinium-‐induced dopaminergic neurotoxicity in rats: implication for immunosuppressive therapy in Parkinson’s disease. Neurosci Bull. 2008 Jun;24(3):133-‐142.98 | P a g e
News Flash, August 31, 2010. European study found that participants in a study who ate a diet that was made up of a diverse mix of fruits and vegetables such as that shown in the photo above, had a lowered risk of a common type of lung cancer. Head of the project was Dr. H. Bas Bueno-de-Mesquita from the cancer epidemiology at the National Institute for Public Health and the Environment in the Netherlands. He explains that a diverse mix means more than just a banana with your cereal and then cooked carrots and peas with dinner. He says to “think kale and spinach; berries and melons; cabbage, cauliflower and eggplant…”There were more than 450,000 adults participating in the nine-year study, from 10 different European countries. Vegetables were divided into categories, like leafy, fruiting or root vegetables, and did not include legumes, potatoes and other tubers. Fruits could be fresh, dried or canned, but not included in the computations were nuts, seeds or olives. When the participants were matched up according to the diversity of their fruit and vegetable intake, it was shown that smokers who ate the greatest variety of fruits and vegetables were 27% less likely to develop squamous cell lung cancer.308 Of course the study concludes with the admonition that quitting smoking is by far the most important way to lower the risk of lung cancer. But we also know that more and more people who have never smoked are getting lung cancer today. So eating a diversity of fruits and vegetables is critical for us all.Of course we know that cooking and canning destroys antioxidants. We have to wonder, if one of the criteria been how many of the fruits and vegetables consumed were raw, most likely that group would have shown an even higher percentage of lung cancer avoidance. In fact, in a similar study conducted by the Roswell Park Cancer Institute in Buffalo, New York, smokers who ate at least 4.5 servings of raw cruciferous vegetables in a month cut their risk for developing lung cancer by up to 50%. This begs the question, would daily consumption of raw cruciferous vegetables cut the risk to nearly 100%? The researchers’ comments were that their findings were consistent with the “biological action of isothiocyanates, phytochemicals that are abundant in cruciferous vegetables.” This study also said that quitting smoking was by far the best way to prevent lung cancer.308 H. Bas Bueno-‐de-‐Mesquita, M.D., M.P.H., PhD, project director. Cancer Epidemiology, National Institute for Public Health, Bilthoven, The Netherlands. Marjorie McCullough ScD., strategic director, nutritional epidemiology, American Cancer Society. Atlanta, September, 2010 Cancer Epidemiology, Biomarkers & Prevention.100 | P a g eIn addition to destroying antioxidants, cooking foods presents another issue. It has been said that cooking and aging have similar biological properties. This is because cooked foods form advanced glycation end products (AGE). Glycation is when a protein and glucose molecule are bound together, forming irreversibly damaged protein structures. Many age-related diseases (for example hardening of the arteries, cataracts, diabetes and neurological diseases) have glycation that occurs within the body in their etiology. Advanced glycation end products in foods become glycotoxins and induce the production of a low-grade, but chronic state of inflammation.309Of course it is difficult to consume 100% raw foods when you’ve eaten cooked foods all your life. Some have done it, but often rely on excessive use of nuts, seeds, honey and even raw grains to be satisfied. So the “trick” is to eat mostly raw, and only consume cooked foods that don’t generate free glutamates or form advanced glycation end products. Generally these foods are low in protein or foods that have been very gently cooked. You have only two food preparation “rules”: Mostly raw (for example two of three meals or 2/3 of every meal) and whatever you cook or consume cooked must be low in protein and/or cooked “gently” (low temperatures, short duration). Of course all foods must not have any free glutamates added.As you embark upon this way of eating you are going to find that preparing healing foods on a daily basis is not so much about cooking, but purchasing fresh organic fruits and vegetables, and preparation. If you’re eating correctly, you’ll be doing far more purchasing, washing, cutting up, combining, dressing and a lot of chewing. But you won’t be doing a lot of cooking. When you do cook, it will be simple – steamed vegetables over rice, dressed with organic extra virgin olive oil and your favorite herbs is a good example of an acceptable cooked meal.You probably used to think of fruits as a big bowl of cereal with a few strawberries on it. Yet what you really need is a big bowl of fresh, organic, raw berries. This also means that a few leaves of lettuce crowned in a commercial salad dressing does not suffice as consuming enough therapeutic vegetables. What is needed every day is an entire bag of organic, raw spinach for the salad, dressed with organic, extra virgin olive oil, which has its own supply of polyphenols. In fact,309 Jaime Uribarri et al. Circulating Glycotoxins and Dietary Advanced Glycation Endproducts: Two Links to Inflammatory Response, Oxidative Stress, and Aging. J Gerontol A Biol Sci Med Sci. (2007 April) 62(4):427-‐433.101 | P a g e
A cereal bowl full of organic blueberries and organic strawberries. A healing afternoon snack.
they are needed. Indeed, most of the time, these chemicals do go where they are not wanted. Hence, we have a list of side-effects as long as your arm for nearly every drug on the market. Some are extremely frightening (“coma” or “death” for example). It is a wonder that the pharmaceutical industry is the multi-billion dollar industry that it is!Thankfully Nature has taken care of the delivery problem. Powerful, healing substances, numbering about 6,000 in just the category of polyphenols alone, not only cross the blood brain barrier when that is where they need to go, but don’t cause the dangerous side-effects that drugs do.The human body consists of between 10 and 50 trillion cells (apparently no-one has stayed up late enough to count them definitively). There aren’t even 7 billion people in the world as of 2010. Can you even fathom how many 50 trillion is? The point is that each and every one of those cells needs protection. Every cell in your body is being attacked from all sides, every day, by “free radicals” (highly reactive, unstable molecules, hell-bent on making themselves stable even if it means destroying your body’s cells). Early on we discussed free radicals being created in Parkinson’s mostly in the form of hydrogen peroxide and peroxynitrite, generated as a result of the damaged mitochondria.Taking a handful of antioxidant pills, while helpful, is not enough. I repeat, approximately 6,000 polyphenols have been studied. Each exert their own mechanism of antioxidant protection. So are your 10-50 trillion cells going to truly get enough protection when you consume a bowl of sugary cereal with pasteurized milk, Big Mac, fries, coca cola, Snickers bar, steak and baked potato, and a piece of pie (sounds like a typical day in the life of the average American to me!) Not only have you added additional oxidative burden from the foods themselves, but nowhere in that list of foods is there even a thimble full of protection for your cells by way of powerful polyphenols. Your 50 trillion cells need more than 50 trillion little “soldiers” standing guard – every day. The only way you’re going to supply that type of protection is by a diet made up, from dusk to dawn, of whole, mostly raw, organic plant foods.Because even nutritionists are often confused, we’ve inserted a chart of where polyphenols fit into the family of phytochemicals. As you read through foods and supplements that are recommended for Parkinson’s, you can refer to the chart to see where that phytochemical fits in. It’s truly fascinating. the chart shows that there are other powerful therapeutic phytochemicals other than polyphenols, and103 | P a g e
Chart Of Phytochemicals
|PHENOLIC COMPOUNDS||Example of Food(s) Rich in This Phytochemical|
|FLAVONOIDS also called Polyphenols|
|Silybin||Milk Thistle Herb|
|Flavan-3-ols also called Flavanols|
|Proanthocyanidin alsocalled procyanidin||Cocoa|
|Isoflavones also called phytoestrogens|
|Coumestans also calledphytoestrogens||Various Herbs|
|LIGNANS also called phytoestrogens|
|Silymarin||Milk Thistle Herb|
|Pinoresinol and lariciresinol||Sesame Seed|
|Cryptoxanthin||Red Bell Peppers|
|Zeaxanthin||Sweet Yellow Corn|
|Astaxanthin||Microalgae and Yeast|
|Lutein||Kale, Spinach, Romaine|
|Perillyl alcohol||Citrus Oils, Some Herbs|
|Phytosterols||Nuts, Seeds, Beans|
|Beta sitosterol||Avocadoes, Nuts, Seeds|
|Omega 3,6,9 fatty acids||Dark Green Leafy Vegetables, Legumes, Nuts|
|Gamma-linolenic acid||Evening Primrose, Borage, Black Currant|
|Oleanolic Acid||Garlic, Cloves|
|Ursolic Acid||Cranberries, Peppermint, Prunes|
|Betulinic Acid||Various Plants Not Commonly Eaten|
|Moronic Acid||Brazilian Propolis|
|Betaxanthins nonglycosidic versions|
|THIOSULPHONATES allium compounds|
|Allyl methyl trisulfide||Garlic, Onions|
|Diallyl sulfide||Garlic, Onions|
|OTHER ORGANIC ACIDS|
|PHYTIC ACID Inositol Hexaphosphate||Seeds|
have been found to be powerfully therapeutic in Parkinson’s disease. Yokozawa showed that green tea catechins protect from peroxynitrite-mediated damage and prevented 3-nitrotyrosine formation (seen in Parkinson’s) better than superoxide dismutase.311 Green tea catechins have also been shown effective in Parkinson’s therapy due to their ability to chelate iron naturally via preventing redox-active transition metal from catalyzing free radical formation.312Green tea catechins is the term to designate the group of catechins in the tea: Epigallocatechin gallate (EGCG), epigallocatechin (EGC), Epicatechin gallate (ECG) and epicatechin (EC). The amount of catechins in a cup of green tea is 60 mg. This varies, of course, because green tea comes from leaves of plants which are subject to growing conditions. Green tea also contains L-theanine, an amino acid, which is a glutamate antagonist.313,314Green tea catechins reduce inducible nitric oxide synthase expression and formation of peroxynitrite. Green tea catechins are also direct scavengers of nitric oxide and superoxide,315 and have stronger protective activity against peroxynitrite-induced oxidative damage than the synthetic peroxynitrite scavenger ebselen316. In addition green tea catechins protect lipid and proteins against oxidative modifications in the brain, all of which are seen in Parkinson’s.317311 Yokozawa T et al. ()-‐Epicatechin 3-‐O-‐gallate ameliorates the damages related to peroxynitrite production by mechanisms distinct from those of other free radical inhibitors. J Pharm Pharmacol (2004) 56:231-‐239.
312 Weinreb O et al. Neurological mechanisms of green tea polyphenols in Alzheimer’s and Parkinson’s diseases. J Nutr Biochem (2004) 15:506-‐516.
313 Shinozaki H, Ishida M. Theanine as a glutamate antagonist in crayfish neuromuscular junction. Brain Res. 1978 Jul 28;151(1):215-‐9.
314 X Di et al. L-‐Theanine Protects the APP (Swedish Mutation) Transgenic SH-‐SY5Y Cell Against Glutamate-‐Induced Excitotoxicity via Inhibition of the NMDA Receptor Pathway. Neuroscience. 2010 Jul 14;168(3):778-‐86.
315 T Nakagawa, T Yokozawa. Direct scavenging of nitric oxide and superoxide by green tea. Good Chem Toxicol (2002) 40:1745-‐1750.
316 T Yokozawa et al. ()-‐Epicatechin 3-‐O-‐gallate ameliorates the damages related to peroxynitrite production by mechanisms distinct from those of other free radical inhibitors. J Pharm Pharmacol (2004) 56:231-‐239
317 Keiko Unno and Minoru Hoshino. Brain Senescence and Neuroprotective Dietary Components. Central Nervous System Agents in Medicinal Chemistry 2007, 7, 109-‐114.109 | P a g eGreen tea catechins also have the ability to chelate excess iron, a factor in Parkinson’s. Of course, when excess iron is lessened, it is prevented from catalyzing free radical formation.318,319Muscadine GrapesMuscadine Grapes contain up to six times more resveratrol, found mostly in the skin and seeds, than any other grape. When high doses of resveratrol was given to fish, their lifespan increased by 56%.320 When high doses have been given to animals in various studies resveratrol inhibited cancer, was a powerful antioxidant, was effective against neuronal cell dysfunction and more. Resveratrol was found to potently reduce LPS-induced PGE2 synthesis and the formation of 8-iso-PGF2a’ a measure of free radical production. Resveratrol reduced the expression of mitochondrial RNA and protein of mPGES-1, a key enzyme responsible for the synthesis of PGE2 by activated microglia, while not affecting COX-2 (the first known inhibitor to do this).321 Unfortunately, these studies have been done only in animals thus far.Resveratrol given to 6-hydroxydopamine (6-OHDA) induced Parkinson’s disease rats showed reduced behavioral, biochemical and histopathological changes caused by free radicals. Immunohistochemical findings in the substantia nigra showed that resveratrol protected neurons from the deleterious effects of 6-OHDA.322Resveratrol inhibits hippocampal cell death induced by trauma or ischemia, and intracellular reactive oxygen species formation.323,324,325318 O Weinreb et al. Neurological mechanisms of green tea polyphenols in Alzheimer’s and Parkinson’s diseases. J Nutr Biochem. (2004) 15:506-‐516.
319 S. Mandel, MB Youdim. Catechin polyphenols: neurodegeneration and neuroprotection in neurodegenerative diseases. Free Radic Biol Med (2004) 37:304-‐317.320 Valenzano DR et al. Resveratrol prolongs lifespan and retards the onset of age-‐related markers in a short-‐lived vertebrate. Current Biology (2006) 16(3):296-‐300
321 Eduardo Candelario-‐Jalil et al. Resveratrol potently reduces prostaglandin E2 production and free radical formation in lipopolysaccharide-‐activated primary rat microglia. Journal of Neuroinflammation. 20074:25.322 MM Khan et al. Resveratrol attenuates 6-‐hydroxydopamine-‐induced oxidative damage and dopamine depletion in rat model of Parkinson’s disease. Brain Research. 2010 Apr 30;1328:139-‐151
323 KT Lu et al. Neuroprotective effects of resveratrol on cerebral ischemia-‐induced neuron loss mediated by free radical scavenging and cerebral blood flow elevation. J Agric Food Chem (2006) 54:3126-‐3131.110 | P a g eResveratrol participates in protecting against brain damage from excitotoxicity326 (primarily excess glutamate activity leading to excess nitric oxide production).Resveratrol protected rat primary neurons against the expression of caspases 3 and 12 and calcium overload thus inhibiting apoptosis.327Resveratrol inhibits oxidized dopamine’s activation of caspases (which leads to the degeneration of dopaminergic neurons), strongly enhancing the expression of the anti-apoptotic protein Bcl-2.328Because studies showed that resveratrol protected against rotenone toxicity (which acts primarily by inhibiting the electron transport chain at complex I of the mitochondria), researchers conclude that resveratrol would be protective in counteracting the mitochondrial production of reactive oxygen species at both the cytosolic (the liquid found inside of cells) and mitochondrial level.329An extract of whole grape exhibited dose-dependent scavenging effects of reactive oxygen species in fruit flies. The extract also inhibited increases of reaction oxygen species when rat liver mitochondria were exposed to a potent lipid oxidant generator. The extract protected enzyme activities of the mitochondrial respiratory electron transport chain, complexes I and II. Researchers concluded that whole324 U Sonmez et al. Neuroprotective effects of resveratrol against traumatic brain injury in immature rats. Neurosci Lett (2007) 420:133-‐137.
325 S Bastianetto et al. Neuroprotective abilities of resveratrol and other red wine constituents against nitric oxide-‐related toxicity in cultured hippocampal neurons. Br J Pharmacol (2000) 131:711-‐720.326 M Virgili, A Contestabile. Partial neuroprotection of in vivo excitotoxic brain damage by chronic administration of the red wine antioxidant agent, trans-‐resveratrol in rats. Neurosci Lett (2000) 281: 123-‐126.
327 QH Gong et al. Inhibition of caspases and intracellular free Ca2+ concentrations are involved in resveratrol protection against apoptosis in rat primary neuron cultures. Acta Pharmacol Sin (2007) 28:1724-‐1730.328 MK Lee et al. Resveratrol protects SH-‐SY5Y neuroblastoma cells from apoptosis induced by dopamine. Exp Mol Med (2007) 39:376-‐384.
329 Unpublished data from K. Aquilano et al. Department of Biology, University of Rome “Tor Vergata”, Via della Ricerca Scientifica, 00133 Rome, Ital. email: firstname.lastname@example.org | P a g egrape extract with resveratrol is a potent mitochondrial protector, and thus a candidate to protect against neurodegenerative diseases like Parkinson’s.330In healthy animals resveratrol significantly dose-dependently increased levels of superoxide dismutase, catalase and peroxidases.331Studies on bioavailability in humans have shown that resveratrol was found to be poorly absorbed in pill form because as it passes through the intestines where it conjugates with glucuronate and sulfonate.332 Even when a very large dose of resveratrol was given (2.5 and 5 grams), the amount that showed up in the blood was far below what would be necessary to do what was seen in the animal studies.333Because of poor absorption, resveratrol was given as a sublingual supplement, to allow for direct absorption through the mucosa of the mouth. When 1 mg of resveratrol (in a 50 ml solution) was held in the mouth for one minute before swallowing, 37ng/ml of free resveratrol was measured in plasma two minutes later. To achieve the same thing with pill form, 250 mg of resveratrol would need to be taken.334 It appears Nature knows best again, as chewing grapes and allowing the juice to bathe the inside of the mouth appears to be the best way to obtain resveratrol!If you do supplement resveratrol, you want to find one that is 99% Trans- resveratrol and free of emodin. Emodin is a “purgative resin” – a chemical naturally present in some plants that can cause diarrhea.Mangoes330 Jiangang Long et al. Grape Extract Protects Mitochondria from Oxidative Damage and Improves Locomotor Dysfunction and Extends Lifespan in a Drosophila Parkinson’s Disease Model. Rejuvenation Research. Vol 12, No 5 (2009) 321-‐331.
331 M Mokni et al. Effect of resveratrol on antioxidant enzyme activities in the brain of healthy rat. Neurochem Res (2007) 32:981-‐987.332 Walle T et al. High absorption but very low bioavailability of oral resveratrol in humans. Drug Metabolism and Disposition. (2004) 32 (12):1377-‐1382.
333 Boocock DJ et al. Phase I dose escalation pharmacokinetic study in healthy volunteers of resveratrol, a potential cancer chemopreventive agent. Cancer Epidemiology, Biomarkers & Prevention (June 2007) 16(6):1246-‐1252.
334 Asensi M et al. Inhibition of cancer growth by resveratrol is related to its low bioavailability. Free Radical Biology & Medicine (August 2002) 33(3):387-‐398.112 | P a g eMangoes contain mangiferin, a polyphenol studied as an antioxidant for Parkinson’s. Mangiferin was shown to reverse oxidative stress likely through its scavenging activity against reactive oxygen species. When Parkinson’s is induced in lab animals using the toxin MPTP, mangiferin protects against cytotoxicity.335 Mangiferin crosses the blood-brain barrier which is absolutely essential if it is to be of any use in Parkinson’s.336GarlicSulfur compounds found in garlic have been found to protect against MPTP toxicity.337SoybeansSoybeans contain a substance called genistein that has been found to protect dopaminergic neurons by inhibiting microglial activation (inflammation in the brain). Genistein is the primary soybean isoflavone. In a “concentration- dependent” manner genistein counteracted lipopolysaccharide-induced decrease in dopamine uptake and loss of tyrosine hydroxylase-immunoreactive neurons.338 Biochanin A, also in soy also protects dopaminergic neurons against lipopolysaccharide-induced damage through inhibition of microglia activation, and thus proinflammatory factors.339It is extremely important to make it clear here that soybeans must only be eaten in their whole, unprocessed state. When soybeans are processed in any way, high amounts of free glutamates are generated. Soybeans can be healthful whole, but are disease contributing in their processed state.335 Amazzal L et al. Mangiferin protects against 1-‐methyl-‐4-‐phenylpyridinium toxicity mediated by oxidative stress in N2A cells. Neurosci Lett (2007) 418:159-‐164.
336 Martinez G et al. Effect of Mangifera indica L. extract (QF808) on protein and hepatic microsome peroxidation. Phytother Res (2001)15:581-‐585.337 Liu KL et al. DATS reduces LPS-‐induced iNOS expression, NO production, oxidative stress, and NF-‐ kappaB activation in RAW 264.7 macrophages. J Agric Food Chem (2006) 54:3472-‐3478.
338 Xijn Wang et al. Genistein protects dopaminergic neurons by inhibiting microglial activation. NeuroReport Vol 16 No 3 (February 2005) 267-‐270.339 Han-‐Qing Chen et al. Biochanin A protects dopaminergic neurons against lipopolysaccharide-‐induced damage through inhibition of microglia activation and proinflammatory factors generation Neuroscience Letters 417 (2007) 112-‐117113 | P a g eTomatoesTomatoes contain carotenoids, like lycopene which have been found to counteract mercury’s toxic effects on cells, in particular the release of cytokines like TNFa.340Diagnostic TestsMercury TestingYou cannot test for mercury in hair341 or blood. Mercury becomes lodged in the brain, bone and other tissue and won’t be discerned by typical testing.Mitochondrial Dysfunction TestsTypical tests screen for disorders of oxidative phosphorylation342. Normal results do not exclude a mitochondrial disorder, however.Fasting Blood Test Complete blood count, comprehensive metabolic panel, creatine kinase, Vitamin B12, methylmalonic acid, leukocyte CoQ10, acylcarnitine profile, ammonia*, lactic acid*, pyruvic acid*, plasma amino acids. [*prone to sample handling/acquisition errors]Urine Test Taken As First Morning After Fasting pH, organic acids, amino acids, acylcarnitine profile.Red Flags increased lactate/pyruvate ratio, decreased serum bicarbonate, increased CK or AST (suggestive of mild metabolic muscle disorder), decreased carnitine, short chain dicarboxylic fatty acids seen on urine organic acid screen (suggests mitochondrial beta oxidation defect), urine organic acids also showing increased fumarate, malate, 3-methylglutaconate, increase alanine relative to lysine* (more stable surrogate marker of pyruvic acid seen on plasma and urine amino acid panels), increased glycine, or proline. [*alanine/lysine ratio is believed by some340 Zefferino R et al. Mercury modulates interplay between IL-‐1beta, TNFalpha, and gap junctional intercellular communication in keratinocytes: mitigation by lycopene. J Immunotoxicol 2008 Oct;5(4):353-‐360.
341 AS Holmes, BE Haley et al. Reduced Levels of Mercury in First Baby Haircut of Autistic Children. International Journal of Toxicology (2003) 22:227-‐285.342 Jon S. Poling MD, PhD. Mitochondrial Disorders and Autism. Athens Neurological Associates. July 12, 2008.114 | P a g eexperts to be the best non-invasive indicator of oxidative phosphorylation disorder; however, unless specifically calculated (normal ratio 1.5 to 2.5) by the ordering physician, the report will come back as normal from the reference laboratory on routine amino acid reports.]MitoSciences sells Monoclonal Antibody Tests, Enzyme Activity Assays, Protein Quantity Assays, Multiplexing Arrays and Enzyme Purification Kits all for the purpose of testing for Complex I dysfunction. 343Elevated Inflammatory Cytokines tumor necrosis factor (TNFa) and IL-6 are the best evidence of neuroinflammation in Parkinson’s disease patients.344Clinical Mitochondrial Therapies [ ] DMSA For Mercury ChelationDMSA (Dimercapto succinic acid) is a sulfhydryl-containing, water-soluble, non- toxic, orally administered metal chelator because it has a high affinity for metals like mercury because of the sulfhydryl group, just like that found in the mitochondrial membrane. However, DMSA has been shown to not cross the blood-brain barrier. In addition, in people with compromised kidney function, DMSA (and many other chelating agents) might redistribute the mercury, dropping mercury back into the body when the kidneys aren’t able to properly excrete the mercury quickly enough. As it also so happens, mercury is toxic to all organs, especially the kidneys.345[ ] Intravenous GinkgoResearchers administered 8 mg/kg “bilobalide” to rats and found a significant level of bilobalide in both plasma and brain.346343 MitoSciences, 1850 Millrace Drive, Suite 3A, Eugene, Oregon 97403. (541) 284-‐1800. email@example.com.
344 H. Wilms et al. Inflammation in Parkinson’s diseases and other neurodegenerative diseases: cause and therapeutic implications. Curr Pharm Des (2007) 13:1925-‐1928.
345 Rudolfs K. Zalups. Molecular Interactions with Mercury in the Kidney. Pharmacological Reviews. March 1, 2000. Vol 52. No 1. 113-‐144. firstname.lastname@example.org
346 Madgula VL et al. Intestinal and blood-‐brain barrier permeability of ginkgolides and bilobalide: in vitro and in vivo approaches. Planta Med 2010 Apr;76(6):599-‐606.115 | P a g e[ ] Methylcobalamin, Folate and B6
348 Orozco-‐Barrios CE et al. Vitamin B12-‐impaired metabolism produces apoptosis and Parkinson phenotype in rats expressing the transcobalamin-‐oleosin chimera in substantia nigra. PLoS One (Dec 21, 2009) 4(12):e8268.349 GA Qureshi et al. Is the deficiency of vitamin B12 related to oxidative stress and neurotoxicity in Parkinson’s patients? CNS Neurol Disord Drug Targets (Feb, 2008) 7(1):20-‐27.116 | P a g e
mitochondria are a-tocopherol (vitamin E) and ubiquinol (Coenzyme Q10). In a healthy body, these more familiar antioxidants are very good at getting rid of lipid peroxyl radicals, thus preventing the peroxidation of lipids (fats). One lipid needing major protection is cardiolipin, the major phospholipid on the mitochondria inner membrane. Cardiolipin is highly susceptible to peroxidation because it is highly unsaturated, which means it has many sites on its molecule which are easily oxidized.350 We know that cardiolipin is associated with cytochrome c. When cardiolipin is oxidized, cytochrome c is released through the outer mitochondrial membrane, something also seen in Parkinson’s.351When cytochrome c is released from the mitochondria into the cytoplasm, it initiates a cascade of events involving caspase 9, 3 & 7 (proteases which are involved in apoptosis) known to do damage to components within the cell352,353 all leading to what is called the intrinsic mitochondrial pathway of apoptosis. Excessive depletion of ATP results in cell death. Of course, it is the death of dopaminergic neurons that is what causes the symptoms of Parkinson’s.SS AntioxidantsThe statement is made that mitochondria undergo oxidative damage when reactive oxygen species production exceeds the antioxidant capacity of mitochondria, and this is true. What is missing, however, is that mercury initiates mitochondrial damage, as if mitochondria simply “self destruct” or at least “mysteriously” destruct. This is critically important because current efforts to cure Parkinson’s focus almost solely on the reactive oxygen species being created by a dysfunctional, damaged mitochondria. Therapies need to be aimed in two places: completely eliminating mercury exposure and repair of mitochondria. This is truly a radical shift in focus, but a shift that must be made if we are ever to “cure” Parkinson’s.If simply taking a-tocopherol, CoQ10 and green tea polyphenols were completely effective in humans, we would have cured Parkinson’s long ago. Yet all of these350 Laganiere S, Yu BP. Modulation of membrane phospholipid fatty acid composition by age and food restriction. Gerontology. 1993;39:7-‐18.351 Shidoji Y et al. Loss of molecular interaction between cytochrome c and cardiolipin due to lipid peroxidation. Biochem Biophys Res Commun . 1999; 264:343-‐347.
352 Green DR, Reed JC. Mitochondria and apoptosis. Science. 1998;281:1309-‐1312.
353 Li P, et al. Cytochrome c and dATP-‐dependent formation of Apaf-‐1/caspase-‐9 complex initiates an apoptotic protease cascade. Cell. 1997;91:479-‐489.117 | P a g eantioxidants, and many others have been shown to have powerful effects against the free radicals produced in Parkinson’s models in animals.354 Unfortunately, most substances tested in labs and animals have failed to show benefits in humans. As we discussed earlier, aiming a nutrient at a cell in a Petri dish, almost never translates to the same effect within a living, diseased human body. The reasons are likely too numerous to elaborate upon here, but a few are the blood-brain barrier, “rebound” effects within the human body, and bioavailability of substances, i.e., can the substance do what it is supposed to within the “environment” it finds itself in, in a human body.If we are going to have any effect upon the extreme free radical damage occurring in Parkinson’s, our therapies are going to have to target the mitochondria. This means that the compounds must be able to cross the blood-brain barrier, and the mitochondrial inner membrane.This all brings us to “SS antioxidants” (Szeto-Schiller). The previously mentioned MitoQ is the mitochondrially-targeted peptide antioxidant SS-31. How do these antioxidants work?Because there is a higher concentration of protons outside the inner membrane of the mitochondria than inside the membrane a negative potential of 150 to 180 millivolts is generated across the mitochondrial inner membrane. Therefore, lipophilic (attracted to fats) cations (an ion with more protons than electrons, giving it a positive charge) are highly attracted to the mitochondrial matrix. And indeed, when antioxidants are made into lipophilic cation compounds, they accumulate 100 to 1000-fold in the mitochondrial matrix.355 The mitochondrial matrix contains the mitochondria’s DNA and is where much activity of the mitochondria takes place.How are antioxidants transformed into those that will target the mitochondria? By conjugating it to a lipophilic cation, such as triphenylphosphonium (TPP+).Coenzyme Q10 and Vitamin E, when combined with TPP+ were found to preferentially accumulate in the mitochondrial matrix. MitoVitE was reported to354 Beal MF, Mitochondria, oxidative damage, and inflammation in Parkinson’s disease. Ann NY Acad Sci. 2003;991:120-‐131.
355 Matthias L. Jauslin et al. Mitochondria-‐targeted antioxidants protect Friedreich Ataxia fibroblasts from endogenous oxidative stress more effectively than untargeted antioxidants. The FASEB Journal August 15, 2003. mpm@mrc-‐dunn.cam.ac.uk Michael P. Murphy118 | P a g ebe 800-fold more potent than idebenone (a drug for Alzheimer’s with properties similar to CoQ10) in protecting against glutathione depletion. MitoVitE was 350- fold more potent than trolox (a water-soluble derivative of vitamin E).MitoQ and MitoVitE inhibited H2O2-induced apoptosis in endothelial cells.356In keeping with all natural functions in the body, there is a “homeostasis” within the mitochondria, seeking a proper balance of all substances and functions. In fact, it has been found that more is not better when it comes to these mitochondria- targeted antioxidants. The mitochondria will self-limit the uptake of the antioxidants at concentrations greater than 50 μM (50 micrometres).357 Too high of concentrations of MitoQ, for example, have been shown to cause mitochondrial depolarization.358Excited about the possibilities, a one year, double-blind, placebo-controlled study to assess Mito-Q’s function as a disease-modifier in Parkinson’s was completed in July of 2010. The study was conducted in 13 clinics in New Zealand and Australia, on idiopathic Parkinson’s disease patients who were taken off of all medications. Patients were given MitoQ at 40 or 80 mg or a matching placebo. At the end of the study the researchers concluded that MitoQ did not slow the progression of untreated Parkinson’s according to their UPDRS359 and other measurement scales. When I inquired about MitoQ, I was told, “MitoQ failed”. The researchers surmised that the sample size was too small, or that the outcome measures were inappropriate.360I would like to suggest that with everything that has been outlined in this book, Co- Enzyme Q10 alone would not stop the progression of Parkinson’s. However, I356 Anuradha Dhanasekaran et al. Supplementation of Endothelial Cells with Mitochondria-‐targeted Antioxidants Inhibit Peroxide-‐induced Mitochondrial Iron Uptake Oxidative Damage, and Apoptosis. The Journal of Biological Chemistry. September 3, 2004. Vol. 279. 37575-‐37587.
357 Smith RA, et al. Selective targeting of an antioxidant to mitochondria. Eur J Biochem . 1999; 263 :709-‐ 716 .358 Kelso GF et al . Selective targeting of a redox-‐active ubiquinone to mitochondria within cells: antioxidant and
antiapoptotic properties. J Biol Chem . 2001 ; 276:4588-‐4596 .
359 http://www.mdvu.org/library/ratingscales/pd/updrs.pdf360 Barry J. Snow MD et al. A Double-‐Blind, Placebo-‐Controlled Study to Assess the Mitochondria-‐ Targeted Antioxidant MitoQ as a Disease-‐Modifying Therapy in Parkinson’s Disease. Movement Disorders ahead of publication, August 2010. Correspondence to: Dr. Barry J. Snow, Neurology Department, Auckland Hospital, Auckland, New Zealand. E-‐mail: email@example.com | P a g e