A Current understanding of Parkinson's Book

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NATURAL

REGENERATIVE

THERAPIES FOR

PARKINSON'S

DISEASE

David A. Steenblock M.S., D.O.

Until you know what causes Parkinson's, you

cannot successfully treat it. Combining hundreds

of scientific findings with my own clinical results,

we now have powerful therapeutic approaches to

combat Parkinson's.

Forest-For-The-Trees Publishing San Juan Capistrano, California 2010

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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, wherein 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.

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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.

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One of the first times mitochondrial dysfunction was described was in the 1960s, but the wide range of disorders that would end up having mitochondrial damage as the basis for their pathology is only now becoming apparent in 2010.2

The mitochondria is, as the Orange County article states, the "energy factory" of your cells (see diagram of cell and its mitochondria). Of course you have cells that make up your entire body, and you have neurons which are special messenger types of cells which enable everything that happens in your body to occur. What we're going to see, as we pull together the scientific studies, is that dysfunctional mitochondria create a continuous excess of superoxide free radicals for reasons we shall explore as we continue. Of course a healthy body would neutralize superoxide free radicals with antioxidants.

Mitochondria have a "respiratory chain" that transfers energy across the outer membrane. The chain is made up of Complex I, Complex II, Complex III and

Complex IV. You'll likely want to refer to the chart below as we continue to

discuss more about the mitochondria.

MITOCHONDRIAL ELECTRON TRANSPORT CHAIN

Complex I Complex II Complex III Complex IV

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.

2

John Neustadt and Steve R. Pieczenik. Medication-induced mitochondrial damage and disease. From "Molecular Nutrition & Food Research" Vol 52, Issue 7, July 14, 2008. 780-788.

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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 an

3

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) 33:2416-2426

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excellent 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 protected].

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showing up in neurological diseases as a "glutathione deficiency" . Glutathione depletion doesn't cause mitochondrial impairment. Mitochondrial impairment is caused by the same thing that glutathione depletion is caused by - mercury. We discuss later why this is so.

The aforementioned study also says, "Oxidative stress and mitochondrial impairment, preceding DNA fragmentation, could be early events in the

apoptotic process induced by glutathione depletion." They go on to say that their

data is consistent with the hypothesis that glutathione depletion could contribute to neuronal apoptosis in Parkinson's disease through oxidative stress and

mitochondrial dysfunction. Of course glutathione depletion leaves gaping holes in the body's antioxidant capabilities. Without enough glutathione, the body is

rendered unable to defend against many of the deadly reactive oxygen species

(defined as any species capable of independent existence that contains one or more

unpaired electrons). This includes, for example, H2O2, peroxynitrite, and hydroxyl radical, all of which are spewed out from damaged mitochondria. At this point if we insert into this equation mercury's propensity for doing damage to

mitochondrial membrane and energy systems, depleting glutathione, and

fragmenting DNA, we would create a more accurate assembly of the puzzle pieces to the Parkinson's conundrum. Why is getting to the true core of the problem of utmost importance? Because the truth is, if you want to put out a fire, you have to

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In the quote from the 1987 study (above) these researchers suggest that the effect of the "poison" (mercury) on the translation mechanism associated with brain

mitochondria needs to be studied. These researchers called mercury right, a

poison. Why then is this poison still being injected into humans in 2010? I ask, because we've known about mercury's devastating toxicity since before Christ. More recently, people making felt hats in the 1800's, which used mercury in the process, observed that the "hatters" went mad and had the "shakes" from mercury.6 The 1987 study talks about mercury's inhibitory effect on cytoplasmic protein. Another study shows how mercury binds to proteins and the enzymes that

assemble proteins.7 In Parkinson's and many other neurological diseases enzyme

systems (made of proteins) that protect cells and orchestrate the proper functioning

of cells are damaged. This would include the glutamatergic system, the

dopaminergic system, and the serotonergic system.

Damaged mitochondria would have a deleterious effect in many areas of the body. Recently, researchers studying macular degeneration and cataracts have found extreme oxidative stress and damage to the mitochondria as the basis. So if mercury is damaging mitochondria, what exactly is it that mercury has such an affinity to that exists in proteins, mitochondria and glutathione? Well, here it is, and this is big. In fact, it is the reason why mercury is our number one suspect above all other toxins, and it is this: It is the sulfur compounds (sulfhydryl

group, thiol, mercaptan) plentiful in cells that has an extraordinary affinity to

mercury. As if that's not enough, here's a shocker: This extraordinary affinity of mercury for sulfur has been known since before Christ. As we've mentioned,

6

Koertge HH. The Hazard of Mercury Poisoning: Don't Be a Mad Hatter. J Am Coll Health Assoc. 1965 Apr; 13:551-558.

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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.

"The mode of neurotoxicity of methyl mercury, an environmental pollutant of considerable concern, involves a direct inhibitory effect on cytoplasmic protein synthesizing systems in brains cells." [Dmitrij A. Kuznetsov. Paradoxical Effect of Methyl Mercury on Mitochondrial Protein Synthesis in Mouse Brain Tissue. Neurochemical Research. Vol. 12, No. 8, 1987, pp. 751-753.]

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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.

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For example, some people deliberately intoxicate themselves. Extreme alcohol

consumption damages mitochondria as well.9 In one study "400 ml of ethanol" (alcohol) was used to "remove excess thiol".10

Interestingly, in China, mercury has become one of the main causes of toxic metal pollution in agriculture. They have found that plants with mercury toxicity show a positive correlation with O2

(superoxide radical) and H2O2 (hydrogen peroxide) in leaves. In addition plants poisoned by mercury show increased activity of NADH

oxidase and lipoxygenase, and damage to biomembrane lipids.11 These same phenomena that have to do with the mitochondria, are also seen in Parkinson's and other neurological diseases. This tells the detective in me to put these puzzle pieces together - they're not disparate bits of information.

So where is all the mercury coming from? The diagram to the left shows that amalgams ("silver fillings") still rank as the worst daily source of mercury toxicity. Vaccines are second, even in 2010 when everyone has been pretty much duped into thinking that "thimerosal" (mercury) has been taken completely out. Another fallacy is that some seafood is okay, when the truth is, all seafood has some level of mercury, coming from our polluted waters.

9

Guo R., Ren J. Alcohol dehydrogenase accentuates ethanol-induced myocardial dysfunction and mitochondrial damage in mice: role of mitochondrial death pathway. PLoS One 2010 Jan 18;5(1):e8757.

10

Mathias Brust et al. Synthesis of Thiol-derivatised Gold Nanoparticles in a Two-phase Liquid-Liquid System. J.Chem.Soc.Chem.Commun 1994

11

Zhou ZS et al. Biological detection and analysis of mercury toxicity to alfalfa (Medicago sativa) plants. Chemosphere 2008 Feb;70(8):1500-9.

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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 and

12_{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,}

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how 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-12.15

In 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.17,18

TNFa 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):1108-14

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While we've known about mitochondrial involvement in neurological diseases for a while, it has only been quite recently that its true relevance has been elucidated. In a 2009 study out of the department of chemistry, Central Washington University we read that even iron accumulation seen in Parkinson's is a part of mitochondrial damage:

Different Types Of Parkinson's?

The question with Parkinson's has always been, is it genetic or is it environmental? In 2009 Author Ruben K. Dagda et al in Mitochondrial Kinases in Parkinson's Disease: Converging Insights from Neurotoxin and Genetic Models [May, 2009] states: "Alterations in mitochondrial biology have long been implicated in

neurotoxin, and more recently, genetic models of Parkinsonian neurodegeneration. In particular, kinase regulation of mitochondrial dynamics and turnover are

emerging as central mechanisms at the convergence of neurotoxin, environmental and genetic approaches to studying Parkinson's disease." They go on to say

"...evidence gathered over the last decade implicate a central role for kinase

signaling at the mitochondrion in Parkinson's and related neurodegenerative

disorders." Again, the mitochondria emerges as the "part" damaged/not working and as these researchers state, "Interactions involving" a-synuclein (a protein found primarily in neural tissue), LRRK2 (a protein found mostly in cytoplasm, but also associated with the mitochondrial outer membrane), DJ-1 (also called PARK7, a protein that seems to protect neurons from oxidative stress and cell death) and parkin (a protein encoded by the PARK2 gene, which when mutated has been linked to early-onset Parkinson's), are involved not causal.19

19

Ruben K. Dagda et al. Mitochondrial Kinases in Parkinson's Disease: Converging Insights from Neurotoxin and Genetic Models. May 6, 2009.

"Our finding together with reports on iron accumulation in degenerative diseases highlight the importance of developing mitochondrial-targeted antioxidants for the

therapeutic intervention of diseases associated with mitochondrial dysfunction and oxidative stress." They say that hydroxyl radical levels occur in mitochondria under

oxidative stress and hydroxyl radical levels can be modulated with antioxidant enzymes and iron ligands. [Thomas C Mackey MM et al Hydroxyl radical is produced via the Fenton reaction in submitochondrial particles under oxidative stress: implications for diseases associated with iron accumulation. Redox Rep 2009;14(3):102-8.]

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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 Mitochondria

When 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.

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In "biology class" perhaps you learned that mitochondria are the "batteries" of cells. But the mitochondria is so much more. Indeed, the mitochondria reside inside of the body's cells. Mitochondria generate adenosine triphosphate (ATP). ATP transports chemical energy within cells for metabolism, which is why your teacher told you mitochondria are the "batteries" of your cells. But the

mitochondria also orchestrate various biosynthesis pathways, the regulation of

calcium homeostasis and apoptotic signaling (telling cells how and when to die).

The mitochondria are also responsible for oxidative phosphorylation, lipid

metabolism, tricarboxylic acid cycle, and iron-sulfur cluster formation.

Diseases known to be caused by mutations in some of the genes associated with the mitochondria, like Charcot-Marie-Tooth subtype 2A and autosomal dominant optic atrophy have been known for some time. But recent studies have shown that dysfunctional mitochondrial fission and fusion (these control the shape and function of the mitochondria) is involved in Parkinson's disease.21

But it's not just Parkinson's that has mitochondrial damage and dysfunction and resultant extreme oxidative stress now suspected at its core. Add to the list

schizophrenia, bipolar disease, dementia, Alzheimer's disease, epilepsy, migraine headaches, strokes, neuropathic pain, ataxia, transient ischemic attack, cardiomyopathy, coronary artery disease, chronic fatigue syndrome, fibromyalgia, retinitis pigmentosa, diabetes, hepatitis C, and primary biliary cirrhosis1. One would think that if we combine the fact that the cause of each of these continues to baffle, the treatment continues to be hit or miss, and that nobody

is being cured - with what we now know about mitochondrial

dysfunction, and mercury's role, that perhaps we could finally address the true underlying cause of these diseases to both prevent and find corrective therapies. Speaking of mitochondrial

fusions, this is thought to be

involved in the transport of

mitochondria along microtubules. Microtubules are one of the

21

Bingwei Lu, PhD. Mitochondrial Dynamics and Neurodegeneration. Current Neurology and Neuroscience Reports 2009, 9:212-219. [email protected]Commonly Affected Systems in Mitochondrial Disorders

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components of the cytoskeleton which is a cellular "scaffolding" or "skeleton" contained within the cytoplasm of cells. A more recent observation is that fission of mitochondria appears to be important for the correct distribution of

mitochondria along neurites and at synapses.22 Microtubules are mostly made of beta tubulin (a protein), which (you guessed it) is readily damaged by mercury.

Dopamine, One Of Many Neurotransmitters

Parkinson's disease has been attributed to what appears to be a deficiency of

dopamine. It does appear there is not enough dopamine in the Parkinson's brain,

but "deficiency" will likely not turn out to be the best way to describe the problem. Dopamine is one of many neurotransmitters in the body. Neurotransmitters are how neurons connect (synapse) to do the jobs they do in the body, that is, to pass along information from one neuron to another, like a chain reaction, ending up in cells where the information is needed. There are approximately 100 billion neurons in the human brain. Neurons travel throughout the body, of course. For example, the way in which you taste food is by neurons in your taste buds relaying the saltiness, sweetness, sourness, etc. of the food to your brain. When the

"dopaminergic" system of neurotransmitters goes awry, as in Parkinson's, bodily functions suffer.

How does the "dopamine" neurotransmitter work? On the end of one neuron are a collection of neurotransmitters and on the target cell there are places where the neurotransmitters are "received" (receptor sites). There is a space in-between the two cells called the "cleft". The neurotransmitters are released into that cleft ultimately binding to the receptors in the membrane on the "target cell". Voila! They've communicated. This connection was thought to be electrical when neurons were first discovered, but in 1921 German pharmacologist, Otto Loewi, found that neurons communicate by releasing "chemicals" which we now call neurotransmitters.

You may have heard the names of some of these familiar neurotransmitters: amino acid types: glutamate, aspartate, serine, GABA, glycine monoamine types:

dopamine, norepinephrine (noradrenaline), epinephrine (adrenaline),

histamine, serotonin, melatonin others: acetylcholine, adenosine, anandamide, nitric oxide.

22

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Receptors are those sites on cells that bind to neurotransmitters to complete the

communication between cells. People have made the analogy of the

neurotransmitter being the "key" and the receptor being the "keyhole". The receptor is actually a protein molecule imbedded in either the outer membrane of the cell or in the cytoplasm within the cell. A molecule that attaches to a receptor is given a general categorical name of ligand. The ligand can be a short protein, a neurotransmitter, a hormone, or even a toxin or a drug. Many drugs, in fact, are created for the purpose of attaching to specific receptor sites in an attempt to alter a disease course. Every cell has many receptors of many different kinds. Most receptors "receive" more than one ligand. Just one example, is that the receptor site for vitamin C also receives glucose (a consideration when we give children sugary drinks instead of fresh fruits with vitamin C).

Neurotransmitters are classified not by how they look, but how they behave.

Therefore, if they behave differently in other functions in the body, they can become something other than a neurotransmitter, for example, they can also behave as hormones (defined as a chemical released by a cell in one part of the body, that sends out messages that affect cells in other parts of the body.) The neurotransmitter glutamate is known to modulate the neurotransmitter

dopamine's release.23,24 This is important, because food manufacturers are putting free glutamates in processed foods by about a million tons worldwide annually.25 This issue of excess glutamate is very important because when there is

mitochondrial damage there is a resultant deficiency in the enzyme needed to process or recycle glutamate. Thus, glutamate is known to build up and become excitotoxic (causing neurons to synapse literally "to death") in neurological diseases like Parkinson's.

I started off by saying that "deficiency" of dopamine may not appropriately

describe what is going on in Parkinson's. As I began to discover in my research, it appears most likely that dopamine isn't deficient, but is being oxidized by all of the reactive oxygen species being generated due to mitochondrial damage. We'll

23

PJ Roberts et al. Effects of L-glutamate and related amino acids upon the release of [3H] dopamine from rat striatal slices. Brain Research 157 (1978) 391-395.

24

PJ Roberts et al. Stimulatory effect of L-glutamate and related amino acids on [3H]dopamine release from rat striatum: an in vitro model for glutamate actions. J Neurochem 32 (1979) 1539-1545.

25

According to Leo Hepner, president of the U.K. consulting firm L. Hepner & Associates, which specializes in fermentations and biotechnology.

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discuss that more later. In addition, mitochondrial-generated reactive oxygen species does damage to the glutamatergic system.

Excitotoxicity: Glutamate & Nitric Oxide

Glutamate is also a neurotransmitter. Researchers studying the mitochondria and its relationship to amyotrophic lateral sclerosis (Lou Gehrig's disease) say that in the central nervous system, glutamate is the principle stimulatory

neurotransmitter, and neuronal mitochondria play an important role in glutamate's metabolism, as well as in the inhibitory neurotransmitter GABA (where glutamate "excites" neurons, GABA "calms" them down). GABA, in fact, is the primary inhibitory transmitter in the brain. Many tranquilizer drugs act by enhancing the effects of GABA. Glycine is the primary inhibitory transmitter in the spinal cord. Excessive stimulation of glutamate receptors is associated with neurotoxicity and the further generation of reactive oxygen species, including excess nitric oxide. 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.26

Glutamate

Glutamate 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 + H2O

26

Alexander Panov MD, PhD Senior Scientist, Mitochondrial Biology Group, Carolinas

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Note the need for ATP. The majority of ATP production takes place in the

mitochondria27 as we've previously discussed, which we now know is damaged in Parkinson's. The result is excess glutamate buildup, and excitotoxicity, along with excess nitric oxide. Studies clearly show that glutamate exposure to neurons

negatively affects the mitochondrial respiratory chain (Complex I), depletes

glutathione and increases nitric oxide.28 To this day, the relationship of mitochondria to glutamate and nitric oxide seems to come under the question, "which came first the chicken or the egg". In case you didn't hear, in 2010,

scientists decided that the chicken had to have come first because the egg can only be formed because of a protein found in the chicken's ovaries. Thus, the egg can only be created inside of the chicken. With regard to glutamate and nitric oxide's relationship to the mitochondria, let us hope that researchers will soon come to the appropriate conclusion that mitochondrial damage comes first. This is important, because it is the mitochondria that needs our immediate attention, and not so much glutamate or nitric oxide.

Note in the above study the words "under physiological conditions". This means under normal, healthy bodily conditions. They do say that under pathological conditions there is elevated glutamine and ammonia (these occur as byproducts when glutamate is recycled). They make note of this occurring in conditions of liver failure. Mercury damage also leads to the endogenous production of excess

excitotoxic glutamate and nitric oxide. In fact, research has shown that

glutamine synthetase, the enzyme that is necessary to convert glutamate back to

glutamine (for recycling) in astrocytes (are a type of glial cells in the brain,

27

Lodish H. et al. Molecular Cell Biology 5th Ed. (2004) New York WH Freeman ISBN 9780716743668.

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.

Under physiological conditions, astrocytes take up L-glutamate from the synaptic gap, metabolize it to L-glutamine and return it to neurons, where L-glutamine is metabolized to L-glutamate and stored in neurotransmitter vesicles. However, under pathological conditions, such as hepatic failure, L-glutamine and ammonium are elevated globally in the brain. [Svoboda N, Kerschbaum HH. L-Glutamine-induced apoptosis in microglia is mediated by mitochondrial dysfunction. Eur J Neurosci 2009 Jul;30(2):196-206.]

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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.36

For 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. For

29

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.

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centuries 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.37

37

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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

38

Bittigau P, Ikonomidou C. Glutamate in neurologic diseases. J Child Neurol. 1997 Nov;12(8):471-85.

39

Andreas Plaitakis MD et al. Neurological disorders associated with deficiency of glutamate dehydrogenase. Annals of Neurology. Vol 15, Issue 2. 144-153. October 7 2004.

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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.40

Another 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.

Nitric Oxide

Nitric oxide is a molecule in the body made up of one atom of nitrogen and one atom of oxygen. It is a "free radical" because it has an unpaired electron looking for a mate. Nitric oxide is actually an important "messenger molecule" because it is highly reactive, and can thus react in some positive functions throughout the

40

Kossoff EH, et al.. A modified Atkins diet is effective for the treatment of intractable pediatric epilepsy. Epilepsia. 2006;47:421–424

41

Marchetti et al. Glucocorticoid receptor-nitric oxide crosstalk and vulnerability to experimental parkinsonism: pivotal role for glia-neuron interactions. Brain Research Reviews. 48 (2005) 302-321.

When the production of nitric oxide is abundant and uncontrolled, it results in damaging effects mainly mediated by its reactive species. In Parkinson's disease, nitric oxide increase is caused either by over expression of nitric oxide synthases or by other mechanisms, including glutamate excitotoxicity. The latter event causes the raise of intracellular calcium levels, which in turn increases nNOS dephosphorylation and its enzymatic activity. Nitric oxide reacts with superoxide anion formed during dopamine metabolism thus generating peroxynitrite that is considered one of the main damaging molecules in dopaminergic neuronal cells. [Katia Aquilano et al. Role of Nitric Oxide Synthases in Parkinson's Disease: A Review on the Antioxidant and Anti-inflammatory Activity of Polyphenols. Neurochem Res (2008) 33:2416-2426.]

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body. 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 also

42

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.

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know that glial cells produce excessive levels of nitric oxide in Parkinson's disease which would also be neurotoxic for dopaminergic neurons.46

Thus, 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.48,49

Protein modifications seen in Parkinson's are caused by nitric oxide through

nitrosylation and nitration. For example, during nitration, a nitro (-NO2) group is

46

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. 26

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added onto tyrosine to form nitrotyrosine. Increased nitrotyrosine was detected in the substantia nigra of in vivo models of Parkinson's.50

In 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.51,52

S-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.54

In 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.55

Nitric 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.57,58

50

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

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All 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 toxic metabolites, this would indicate a need to inhibit excess nitric oxide, not attempt to stimulate it.59 Yet some studies recommend therapies or supplements to increase nitric oxide. Creatine and arginine would be examples of two supplements in part aimed at increasing nitric oxide. Again, nitric oxide overproduction is what research has shown to be the event significantly contributing to death of

dopaminergic neurons, via oxidative damage on cellular lipids, proteins and DNA.60,61

A 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.

A Current understanding of Parkinson's Book

NATURAL

REGENERATIVE

THERAPIES FOR

PARKINSON'S

DISEASE

David A. Steenblock M.S., D.O.

Until you know what causes Parkinson's, you

cannot successfully treat it. Combining hundreds

of scientific findings with my own clinical results,

we now have powerful therapeutic approaches to

combat Parkinson's.

A CURRENT UNDERSTANDING OF PARKINSON'S

TABLE OF CONTENTS

MITOCHONDRIAL DAMAGE AND PARKINSON'S