| Main Menu   | Home  |  About Us  |  What's New | FAQSite Search   | Contact Us   |  Books  | Privacy Policy | 

Scientific Rationale for EDTA Chelation Therapy in Treatment of Atherosclerosis and Diseases of Aging

Elmer M. Cranton, M.D.

Copyright © 2013 Elmer M. Cranton, M.D.

This paper is based on an earlier version written in collaboration with James P. Frackelton, M.D., and has been extensively rewritten and updated in 2013. A search of the current literature reveals no contradictions to that former publication and adds much that is new. Updated references have been inserted numerically in order of appearance in the text but end with alphabetical designators.

ABSTRACT: The free-radical theory of aging provides one scientific explanation for benefits reported following administration of intravenous disodium EDTA chelation therapy, but EDTA  has many other actions. The emerging cell-senescence model of aging, and studies of apoptosis (programmed cell death), give us further scientific rationales. Compromised and ischemic tissues accumulate toxic levels of otherwise essential metallic nutrients. Infective micro-microorganisms are shielded against body defenses by biofilms, which are bound in place by polyvalent metallic cations. EDTA is able to rebalance concentrations of metallic cations among cells, organs, and fluids—reactivating metalloenzymes that require metal cofactors in proper locations and in proper ratios for healthy function. EDTA is able to reduce metals that have reached excessive levels. EDTA is able to remove metallic catalysts of free radical proliferation. Any proposed mechanism of action must also explain why full benefit is delayed for several months after treatment to fully occur and why improvement persists long thereafter.

Introduction

Chelation therapy with intravenous disodium ethylene diamine tetraacetate (EDTA) for the treatment of atherosclerosis began more than four decades ago. Many published studies confirm safety and effectiveness.(1-89) It has been reported consistently in widespread clinical practice. More than one million patients have received more than twenty million infusions without serious adverse effects—when administered following the approved protocol. [LINK] Many years ago, reports of kidney damage and other adverse effects resulted from excessive doses of EDTA, infused too rapidly (greater than 50 mg/Kg/day, infused more rapidly than 16.6 mg/min).

Excessive dose-rates of infusion, especially in the presence of preexisting kidney disease or heavy metal toxicity, were responsible for occasional reports of nephrotoxicity.(64-74) No such adverse side effects have been reported when the currently approved protocol has been correctly followed.(72,73,74)

Research with laboratory animals provides further support for the effectiveness of EDTA chelation therapy.(77-83)

There has never been a valid scientific study of EDTA chelation that did not show effectiveness, although there have been reports in which positive data were misleadingly interpreted as negative. Reports of negative or adverse results from EDTA chelation following the currently approved protocol have been either editorial comments and letters to the editor written by uninformed critics of this therapy, often by cardiovascular surgeons who freely admit their bias.(75,84-89)

Positive Data Misleadingly Reported as Negative

In the last ten years, a small cluster of studies has appeared in the medical literature purporting that EDTA chelation is not. Although highly flawed, those studies in actuality provide good support for chelation. Their negative conclusions are contradicted by their data.

The Danish Study

The most controversial and oft cited study of that type was done in Denmark. It was the handiwork of a group of Danish cardiovascular surgeons, who openly admitted their bias against chelation. Results of that study were published in two medical journals, the Journal of Internal Medicine and the American Journal of Surgery. The surgeons’ adverse conclusions were also widely publicized in the news media.(84,85)

The surgeons followed 153 patients suffering from intermittent claudication. The patients had such severely compromised circulation in their lower extremities that walking a city block or less would cause them to stop with pain. An endpoint measured for this study was their maximal walking distance (MWD)—the very longest distance that they could walk before pain of claudication brought them to a halt. Patients were equally divided into an EDTA group and a placebo group. In the pre-treatment phase, the EDTA group averaged walking 119 meters before pain stopped them; the placebo group was less limited at the outset and averaged 157 meters.

Treatment was either 20 intravenous infusions of disodium EDTA or 20 infusions of a simple salt solution, depending on their group. Although the study was purportedly double-blinded (neither patients nor researchers were supposed to know who received placebo and who received EDTA), the researchers later admitted that they broke the code well before the post-treatment final evaluation.

Both groups showed improvement, and the investigators concluded that the improvement was not statistically significant. This Danish study turned many people against chelation; but, in rather short order, the integrity of the study was called into question. It was learned that the researchers had violated their own double-blind protocol. Not only did they themselves know before the end of the study who was receiving EDTA and who placebo, they had also revealed this information to many of the test subjects. Before the study was over more than 64 percent of the subjects were aware of which treatment they had received. This was highly questionable from an ethical and scientific standpoint.

One important aspect of the Danish study is the startling fact that the patients who were given EDTA were much sicker than the patients treated with a placebo. Therefore, the improvements the EDTA group made were harder earned and more significant. The researchers (who had candidly admitted that they undertook the study to convince the Danish government not to pay for chelation) either never noticed that aspect or felt reluctant to reveal it. The evidence is seen in the pre-treatment MWDs: 119 meters for the EDTA group and 157 meters for the placebo group.

Still more significant were the standard deviations. The plus or minus 38 meters SD for EDTA patients versus the plus or minus 266 meters SD for the placebo group represents an enormous variation in walking capacity that is heavily biased in favor of the placebo group. Those standard deviations show that some placebo patients must have walked half a mile before stopping. The placebo group’s claudication was therefore markedly less severe, and the EDTA group was much more severely diseased. The design of the study was obviously biased against EDTA chelation from the outset.

Yet, when the six-month study was completed, the mean MWD in the EDTA group increased by 51.3 percent, from 119 to 180 meters, while the mean MWD in the placebo group increased only 23.6 percent, from 157 to 194 meters. The chelation group’s improvement was therefore more than twice as great as the placebo group’s, even though the chelation group was significantly sicker at the outset. This is a positive study, supporting the usefulness of EDTA chelation. The authors’ published negative conclusions are not supported by the data.

The New Zealand Study

Another study—also conducted by vascular surgeons—was done at the Otago Medical School in Dunedin, New Zealand two years after the Danish study. The subjects of this study were also suffering from intermittent claudication. The subjects were divided into two groups, the EDTA group and the control group. The study extended to three months after 20 infusions of either EDTA or a placebo were given. The authors concluded that EDTA chelation had been ineffective. Once again, that conclusion was unsupported by their data.(86,87)

Absolute walking distance in the EDTA group increased by 25.9 percent; while in the placebo group, it increased by 14.8 percent. The difference was not considered statistically significant. The study, however, had only 17 subjects in the placebo group. One of the placebo patients was what the statisticians call an “outlier,” whose walking distance increased by almost 500 meters. Gain in the placebo group would result from inclusion of this individual’s progress in the final data. Without him, the placebo group’s distance as much less compared with the EDTA treated group.

This illustrates the perils of a small study. The 25 percent gain in the EDTA group compared to no gain in the placebo group would have been very significant statistically.

In addition, the New Zealand researchers conceded that improvement in artery pulsatility (pulse intensity) in the EDTA group’s worse leg improved enough to reach statistical significance (p<0.001).

A 25.9 percent improvement in walking is by no means minor and would attract notice if the agent had been a patentable drug. Even that level of improvement is not representative of the much greater improvements claudication patients normally experience after chelation. The below-expected improvement seen in this study can be explained by smoking. Eighty-six percent of the chelated subjects were smokers. Although they were advised to quit smoking when the study began, how many of them actually complied is not known.

The Heidelberg Study

Another study that was carried out with an erroneous negative conclusion is the “Heidelberg Trial,” funded by the German pharmaceutical company Thiemann, AG in the early 1980s. A group of patients with intermittent claudication were given 20 infusions of EDTA and compared with a so-called “placebo” group that was actually given Bencyclan, a pharmacologically active vasodilating and antiplatelet agent owned by Thiemann.

From a practical commercial standpoint, Thiemann’s action was bizarre. If EDTA did well in the trial, Thiemann’s well-established drug could only suffer. Nonetheless, the trial went forward and was reported, with misrepresented data, in 1985 at the 7th International Congress on Arteriosclerosis in Melbourne, Australia.87 Immediately following 20 infusions of EDTA the trial subjects’ pain-free walking distance increased by 70 percent. Patients receiving bencyclan increased their pain-free walking distance by 76 percent. Twelve weeks after the series of infusions was completed, the EDTA patients’ average pain-free walking distance had continued to increase, going up to 182 percent. No further improvement had occurred in the patients receiving bencyclan. Those percentages were made public, although never published.(87)

An informal report from Thiemann mentioned only the 70 and 76 percent figures. Press releases stated that chelation was no better than a placebo, but failed to mention that the “placebo” was a drug that had been proven effective in the treatment of intermittent claudication. Thiemann never released the actual data on which the Heidelberg Trial based its conclusions, but some German scientists who had access to it, and who were disturbed at the deception they were witnessing, chose to reveal the complete raw data to members of the American scientific community.

The complete data showed that four patients in the EDTA group experienced more than a 1,000-meter increase in their pain-free walking distance following treatment. That highly significant data from those four patients mysteriously disappeared before the final results were made public. Thiemann had a legal right under terms of their contract to edit the final results and to interpret the data in any way that suited them. Another analysis of the data, with the four deleted patients included, showed an average increase in walking distance of 400 percent in the EDTA treated group—five times the 76 percent increase of the group receiving bencyclan.(89)

The Kitchell-Meltzer Reappraisal

A dark moment for chelation research occurred in 1963, when Drs. J.R. Kitchell and L.E. Meltzer coauthored an article reassessing their support for EDTA chelation.(75)

Although it was hardly in widespread use in 1963, chelation had not been controversial. Beginning in 1953, Dr. Norman Clarke and his associates at Providence Hospital in Detroit began using EDTA chelation to treat coronary artery disease. In 1956 they treated 20 patients suffering from angina pectoris. Nineteen of the 20 patients who received EDTA had had a “remarkable improvement” in symptoms.1

Soon other physicians became interested, among them Kitchell and Meltzer, at Presbyterian Hospital in Philadelphia. From 1959 to 1963, Kitchell and Meltzer reported good results treating cardiovascular diseases with EDTA. Their early reports were all very positive.(7,10,14)

In April of 1963, shortly after their last favorable report, Kitchell and Meltzer published a “reappraisal” article in the American Journal of Cardiology that questioned chelation’s value.(75)

In that reappraisal, they reported on 10 of the original patients they had treated for cardiovascular disease, plus another 28 patients that were treated subsequently. Patients in this study were all severely ill. The authors state, “. . . we selected ten patients referred to us because of severe angina. The patients had previously been treated with most of the accepted methods, and their inclusion in this study resulted from wholly unsuccessful courses. Each of the patients was considered disabled at the start of therapy.” This was therefore a high-risk group of very sick patients, who had not improved with any other form of therapy.

Seventy-one percent of patients treated had subjective improvement of symptoms, 64 percent had objective improvement of measured exercise tolerance three months after receiving 20 chelation treatments, and 46 percent showed improved electrocardiographic patterns. Kitchell and Meltzer concluded that chelation was not effective because some patients eventually regressed long after treatment. However, considering the poor health of the patients, some eventual worsening would be expected with any treatment. Eighteen months following therapy, 46 percent of the patients remained improved. The results were very favorable, even though the authors’ conclusions were not.

Kitchell and Meltzer’s reappraisal article was largely responsible for termination of hospital-based, academic research into chelation as a treatment for cardiovascular disease. Rather than analyzing the data for themselves, many physicians simply accepted the flawed conclusion at face value. We will probably never know what prompted those early researchers to change their position so abruptly. We can only speculate that it was an unrealistic expectation that the emergence of bypass surgery would be a final solution. At least one of the authors was a cardiovascular surgeon.

Cell-Senescence Model of Aging

The cell-senescence model (sometimes called the telomere theory) of aging is now hypothesized to underlie almost all aspects of age-related diseases, including atherosclerotic cardiovascular disease.(90-92) As cells continuously die and are replaced with daughter cells, accuracy of gene expression progressively deteriorates. Replacement cells, produced by cell division and DNA replication, grow increasingly weaker. With each replication, accuracy is lost and subsequent generations of cells reflect that deterioration.

Telomeres on chromosomes shorten with each successive cell division, eventually becoming spent. After many divisions, cells reach a so-called Hayflick limit; telomeres become fully depleted and those depleted cells lose their ability to divide. Without telomere replacement, cell death results.

There is more to the story than that. Telomere shortening also correlates with cell senescence, leading to gradual but progressive deterioration throughout multiple generations of daughter cells. When cell deterioration is sufficient to cause impairment in organ function, age-related disease results. It was once thought that only the absolute limit of cell division was important, when telomere length was totally depleted. We now know that as cells divide and telomeres shorten, cumulative inaccuracies in DNA replication cause progressive deterioration of gene expression. This is termed cell senescence. Cells divide and heal more slowly with each successive division.

Two types of cells that do not senesce are germ cells and cancer cells, both of which contain telomerase, an enzyme that restores telomeres. They are thus called immortal cells. When other types of cells, such as fibroblasts, are genetically modified in culture to contain telomerase, they also become immortal and do not senesce. Cell senescence is not only reversed, but aging ceases in cells that produce telomerase, even after 400 or more subsequent divisions.(93)

Endothelial cells in blood vessel walls, lacking telomerase, deteriorate with each cell division.94 With progressive telomere shortening, cell division and replacement slows and the healing process is retarded. Replacement cells also become increasingly weaker and defective with each division. Cells that are subjected to frequent trauma and injury divide more frequently and therefore age more rapidly. Endothelial cells become disrupted at points of stress, and divide most frequently at precisely those points where atherosclerosis prevails. When endothelial cells are injured and replaced by adjacent cells, daughter cells are produced to fill in the gap. Damage to endothelia occurs with increased stress at points of bifurcation caused by sheer effects, from hypertension, infection, toxins, tobacco byproducts, hyperglycemia, oxygen radicals, oxidized LDL cholesterol, and autoimmune processes. The cell senescence model therefore accommodates and provides a broadened explanation for known risk factors of atherosclerosis.

Senescent endothelial cells divide at a progressively slower rate and are less effective at healing breaches in arterial walls. Prolonged exposure of denuded subendothelial tissues triggers a cascade of events that encompasses current theories of causation: monocytes and platelets adhere to damaged areas; monocytes transform into macrophages; a variety of trophic factors and mitogenic factors, including cytokines and platelet-derived growth factor, are released locally; smooth muscle cells proliferate; oxidized lipids accumulate in macrophages; and eventually this enlarging plaque calcifies or ulcerates. An important underlying cause of this chain of events is postulated to be the progressive telomere shortening in endothelial cells at points where they are most often called upon to divide. This occurs at arterial sites most subject to damage and therefore to plaque formation.

Cell types that divide frequently throughout life are precisely those cells that show age-associated decline. Children with one type of progeria (rapid aging) are born with short telomeres. These cells quickly senesce and reach the Hayflick limit of cell division. Cells in skin and hair follicles are replaced frequently throughout life. Resulting deterioration is plainly visible to the naked eye and a close estimate of chronological age can be made at a glance. Cells that divide frequently throughout life age more rapidly at predictable rates include chondrocytes, fibroblasts, keratinocytes, microglia, hepatocytes, and lymphocytes. Astrocytes in the continue to divide throughout life and are prominent in the early inflammatory stages of Alzheimer’s dementia. Neurons make up only 10 percent of brain cells.

The cell senescence model leads to the conclusion that cell division and telomere shortening are central to cell senescence, and thus to age-related diseases.

To explain actions of EDTA using the cell senescence model, consider that biochemical pathways in cell division and replication depend on a large number of metallo-enzymes. DNA-dependent RNA polymerase, an enzyme involved at an early stage of cell replication, is zinc-dependent. Other enzymes needed for replication require the entire spectrum of essential nutritional minerals and trace elements. EDTA has its only known effect on those metallic ions—binding, redistributing, and removing them.

Recent data show that intracellular metallic elements accumulate abnormally in diseased tissues. Although essential at low levels, those are all toxic in excess. Ischemic myocardial cells accumulate an excess of trace elements to quite high levels compared with healthy, young control subjects: cobalt increases 500 percent; chromium increases 520 percent; iron increases 400 percent; and zinc increases 280 percent.(95) Metallic trace elements have a narrow margin between normal and toxic levels. Such three- to four-fold intracellular elevations can poison those cells. It is postulated that by rebalancing a normal distribution of metallic elements within the body, EDTA chelation therapy produces its some of benefits.

Apoptosis (Programmed Cell Death)

When intracellular stresses reach a critical threshold, permeability transition pores (PTP) open in mitochondrial membranes and holocytochrome-C is released. The combination of holocytochrome-C with d-ATP then triggers cellular damage and death. That process is termed apoptosis. Cells become increasingly susceptible to apoptosis with each successive DNA replication and cell division, adding support to the cell-senescence model introduced above. When free oxygen radicals reach high enough levels, apoptosis occurs. Formation of a specific protein, called BAX protein, also opens the PTP, and triggers apoptosis.(96,96A,97) Synthesis of BAX protein requires enzymes that contain metals as cofactors. Every step in the process of apoptosis involves metal-dependent enzymes, and it is on those metals that EDTA has its only known action.

Free Radical Causes of Degenerative Disease

The free radical concept helps to explain contradictory epidemiologic and clinical observations and provides an a scientific rationale for use of EDTA in treatment and prevention of atherosclerosis, dementia, cancer, arthritis, and other age-related diseases.(98-107) Detection and direct measurement of free radicals has only recently been possible.(108-110) The field of free radical biochemistry is as revolutionary and profound in its implications for medicine as the germ theory was for the science of microbiology. It has created a new paradigm for viewing the disease process. Recent discoveries in the field of free radical pathology, in combination with the cell senescent model and apoptosis, provide a coherent and elegant explanation reported benefits following EDTA chelation therapy.

Properly administered intravenous disodium EDTA, together with a program of applied clinical nutrition and modification of health-destroying habits, act synergistically to prevent free radical damage.

What Are Free Radicals?

A free radical is a molecular fragment with an unpaired electron in its outer orbital ring, causing it to be highly oxidative, unstable, and to react instantaneously with other substances in its vicinity.(111,112) The half-life of biologically active free radicals is measured in microseconds.100 Within a few millionths of a second, free radicals have the potential to react with and damage nearby molecules and cell membranes, producing an explosive cascade of free radicals in a multiplying effect—a literal chain-reaction of damage.(98, 98A,101,102,113,113A,114)

Free radicals react aggressively with other molecules to create aberrant compounds. Harmful effects of high-energy ionizing radiation (ultraviolet light, x-rays, gamma rays, nuclear radiation, and cosmic radiation) are also caused by the free radicals produced in living tissues when photons of radiation knock electrons out of orbiting pairs.105,115-119 Free radicals in cell membranes produce damaging lipid peroxides, oxyarachidonate, and oxycholesterol products.98, 98A,119-122 Oxidized cholesterol is toxic and contributes to atherosclerosis. Lipid peroxides can lead to chain reactions, accelerating a cascade of damaging free radical reactions. Protection against free radical damage is achieved from dietary and endogenous antioxidants.98, 98A,99,101-103,106,106A,121,123,124,124A Ongoing free radical reactions in normal cellular metabolism occur continuously in all cells of the body and are necessary for health.98-103,105-107,125-128 Mitochondrial oxidative phosphorylation produces free radicals during the production and storage of energy (ATP) from oxygen. These normal and essential free radical reactions are contained and damage is prevented if adequate antioxidant protection is available. The highly reactive free radicals continuously produced within healthy human cells include hydroxyl radicals, superoxide radicals, and excited or singlet state oxygen radicals. They are commonly referred to collectively as “free oxygen radicals,” or more simply as “free radicals.”99-107 When free radicals react in the body they in turn produce other highly reactive molecules, including hydrogen peroxide, lipid peroxide, and other peroxides. Peroxides are also metastable, highly reactive, corrosive molecules and also react rapidly, producing additional organic free radicals in surrounding tissues.111,112

To prevent uncontrolled propagation of free radicals, cells normally contain a dozen or more antioxidant control systems that regulate the many necessary and desirable free radicals present.98-106,106A 109,110,116,121,123,129-134 Those control mechanisms include endogenous enzymes, such as catalase, superoxide dismutase, and glutathione peroxidase. Free radical regulation also depends on nutritional antioxidants such as vitamins C and E, beta-carotene, coenzyme Q-10, and the trace element selenium. In fact, almost all vitamins, including the B vitamins, play a role in antioxidant protection. When functioning properly, antioxidant systems suppress excessive free radical production, allowing oxidative energy metabolism to proceed without cellular or molecular damage. When those control systems are weakened, free radicals multiply out of control, much like a nuclear chain reaction, disrupting cell membranes, damaging enzymes, interfering with both active and passive transport across cell membranes, and causing mutagenic damage to nuclear DNA. This is one cause of cancer.102,109,113, 113A,114,120,121,135,135A,136

Concentration of the free radical control enzyme, superoxide dismutase (SOD), in mammals is directly proportional to life span. Humans have the highest concentrations of SOD. SOD is the fifth most prevalent protein in the human body.101,102 Elephants, parrots, and other long-lived animals also have high levels. Thus, life expectancy seems to be highly dependent on effective free radical regulation.

Nonenzymatic free radical scavengers are stoichiometrically consumed on a one-to-one ratio when neutralizing free radicals. These include beta-carotene (provitamin A), vitamin E, vitamin C, glutathione, cysteine, methionine, tyrosine, cholesterol, some corticosteroids, and selenium. Once neutralized, other vitamins and enzymes are necessary to restore antioxidant activity, but they must all be present in adequate amounts.

Enzymes involved in free radical protection are proteins, but also require metallic co-enzymes. For example, copper, zinc, and manganese are all essential for superoxide dismutase activity; selenium is essential for glutathione peroxidase; and iron is necessary for catalase and some forms of peroxidase. Tens of thousands of different enzymes in the body depend on vitamins, trace elements, and minerals to function. Optimum dietary intake of those nutrients is therefore necessary for protection against free-radical mediated, age-related diseases.

Identifying Free Radicals

Free radicals rarely reach levels high enough for direct analysis.98, 98A,102 Sophisticated instruments that allow us to recognize the importance and extent of free radical damage in tissues have only recently become available. Electron paramagnetic resonance spectroscopy (EPR) is one type of technology now used.109,110 Free radicals can be estimated most easily, and perhaps even more accurately, by analyzing end-products of free radical reactions, using gas chromatography, mass spectroscopy, and high-performance liquid chromatography. Cross-linkages between molecules, damaged collagen, lipid peroxides, oxyarachidonate, oxidized cholesterol, lipofuscin, ceroid, and increased pigment are all caused by free radical reactions. Those substances can be easily measured.98, 98A,101,102,110,137,138

By sifting through the molecular wreckage left in the wake of evanescent free oxygen radicals, it thus becomes possible to indirectly estimate the type and extent of ongoing free radical reactions. For example, free radical damage in the brain and central nervous system (CNS) can be assessed by the rate of cholesterol depletion. Cholesterol is not otherwise metabolized in the nervous system. The only way for cholesterol to decrease in the CNS is through oxidation caused by free radicals. Cholesterol acts as an antioxidant and is consumed in the process.101,102,139,140

Cholesterol Metabolism

As a free radical scavenger, cholesterol is liberally disbursed throughout cell walls and lipid membranes in the body. Contrary to the popular notion that cholesterol is harmful, it actually protects cell membranes—if it has not previously become oxidized in the process of neutralizing a free radical.110,139 Unoxidized cholesterol is one of the body’s important antioxidant defenses. Cholesterol-derived steroid hormones, including glucocorticosteroids, dehydroepiandrosterone (DHEA), pregnenolone, testosterone, progesterone, and estrogen, can all function as free radical scavengers.110 Those substances decline steadily with age, inversely related to the increase of age-associated diseases.

Cholesterol is a precursor to vitamin D. Vitamin D is normally produced in the skin by exposure of cholesterol to ultraviolet radiation from sunlight. Without cholesterol, vitamin D deficiency can occur. Ultraviolet light is a form of ionizing radiation that can also produce free radicals in the skin manifested as sunburn and skin cancer. Unoxidized cholesterol and other antioxidants act to protect the skin.

Total body cholesterol (approximated by measuring blood levels of cholesterol) is derived primarily from cholesterol synthesis within the liver, not from dietary intake.102 Blood cholesterol levels increase with free radical stress. Elevation of blood cholesterol may be an indicator, not a cause, of excessive free radicals and increased risk of atherosclerosis and apoptosis. As cholesterol becomes oxidized, in the form of low-density lipoprotein (LDL) cholesterol, LDL receptor sites in the liver and elsewhere are altered, causing increased hepatic synthesis of endogenous cholesterol. In this context, an increase in cholesterol is thus a desirable physiologic response to free radical stress. Cholesterol is synthesized in the body as needed, and the need is greater to protect those at risk. Where atherosclerosis, cancer, and other free radical mediated diseases are epidemic, blood cholesterol levels commonly increase with age.

After encountering and neutralizing a free radical, cholesterol is oxidized to become LDL cholesterol. As oxidized LDL, it is toxic to blood vessel walls. If antioxidant protection is diminished, or if free radical production exceeds the threshold of tolerance, oxidized LDL cholesterol increases and contributes to atherosclerosis.

A recent multi-country study in Europe, funded by the World Health Organization, showed low blood levels of vitamin E are statistically 100 times more significant as a predictor of coronary heart disease than are high blood levels of cholesterol.141 In another report, all published autopsy studies that attempted to correlate the extent of atheromatous plaque with levels of blood cholesterol were reviewed. Surgical specimens removed at the time of bypass surgery were also analyzed. No significant correlation was found between blood cholesterol levels and the severity of atherosclerosis.142

Less than one half of one percent of the population has a hereditary trait for dangerously high cholesterol. People with that genetic disease have cholesterol blood levels above 400 mg/dL. They commonly suffer premature death from atherosclerosis despite aggressive pharmacological therapy to lower blood cholesterol. The statements in this section do not apply to such people.

Free radicals oxidize cholesterol into a variety of break-down products.98,100,101,102,110,143,144 Oxidized cholesterol is bound selectively to low-density lipoproteins, referred to as LDL cholesterol, while unoxidized (antioxidant) cholesterol is predominately bound to high-density lipoproteins, HDL cholesterol.101,102 Oxidized LDL is toxic to cells.

Laboratory research has demonstrated that both EDTA and the antioxidant glutathione prevent LDL cholesterol from becoming toxic.143

Oxidized forms of cholesterol possess varying toxicities.98, 98A,102,143-146 Some of those substances have abnormal vitamin D-like activity, which can cause abnormal calcium deposits locally. 144 Free radicals also cause tissue calcification by damaging the integrity of cell membranes, causing leaks in cell walls, damaging enzymatic cell-wall transport pumps. If the calcium pump is weakened, or if cell wall integrity is damaged, the calcium pump becomes unable to remove calcium as it leaks in. Intracellular calcium accumulates, causing malfunction and eventually cell death. X-rays of older people commonly show dense calcium deposits in soft tissues that do not normally have that bony appearance. A similar weakening of the sodium pump in cell walls allows an increase of intracellular sodium, leading to swelling of the cell and eventual cell lysis.

Dietary restrictions of cholesterol and prescription drugs to reduce blood cholesterol have, in some ways, been counterproductive because the antioxidant role of cholesterol has not been widely recognized. Natural, unoxidized cholesterol is widely dispersed in cell membranes as a protective factor against atherosclerosis, cancer, and other free radical-induced diseases. In this form, cholesterol is not the harmful substance we have been told. Cholesterol is a fat, and dietary cholesterol is consumed in fatty foods. Restriction of dietary cholesterol necessarily results in simultaneous reduction of total dietary fats. Research studies that allegedly show benefit from low-cholesterol diets may have only reflected benefit from reduction in excessive fat and the accompanying lipid peroxides and oxidized cholesterol, as often occur in processed foods.

It is not widely known that cholesterol-lowering drugs also have antioxidant activity, and antiplatelet activity.147 Those drugs also produce significant toxicity and cost much more than antioxidant nutritional supplements.

A statistical correlation has been reported between low blood cholesterol and increased risk of cancer.148 Cancer is caused in part by free radical damage to DNA. Free radicals can act as both primary initiators and subsequent promoters of malignancy. If antioxidant protection for nuclear membranes and DNA is lost, an important defense against mutation and cancer is also lost. Diets high in rancid fats are rich in lipid peroxides and increase the risk of cancer.98, 98A,101,102 However, if antioxidant defenses are reinforced, dietary fats can be protected against damaging oxidation and rendered useful for energy.

Free radicals cause damage by oxidation. Fats, especially unsaturated fats, are especially susceptible to oxidation, in the same way that cooking fats and oils can easily catch fire on the stove. They ignite (oxidize) easily and burn vigorously. All cells and intracellular organelles are enveloped in easily oxidized layers of unsaturated fat. Damaging oxidation of fatty cellular and intracellular membranes by free radicals can be prevented in three ways:

1) By “fire-proofing” lipid membranes with nutritional anti-oxidants;

2) By depriving the fire of fuel by partially restricting dietary fats; and

3) By removing metallic catalysts of free radical proliferation with EDTA chelation therapy (as described in detail below).

These three strategies used together act synergistically to reverse and slow diseases of aging.

Homocysteine Metabolism

Homocysteine can contribute to atherosclerosis in several ways. The higher the homocysteine level, the greater the statistical risk. Free radical production and oxidative stress occur during homocysteine metabolism.149-153 Homocysteine is a metabolite of methionine, and is oxidized by free radicals to become homocysteic acid, a potent stimulator of cell growth and multiplication.154 In this way, oxidative breakdown of homocysteine can induce proliferation of smooth muscle cells in arterial walls and promote growth of atherosclerotic plaques.155 Metabolic pathways of homocysteine that cause damage to blood vessel walls involve hydrogen peroxide, superoxide radical, and inhibition of glutathione peroxidase.156 Homocysteine also increases the tendency for blood clotting.157 In addition, homocysteine can increase the oxidation of cholesterol, which then becomes bound to small dense LDL particles and is taken up by macrophages to become foam cells in plaque.158

Damage to blood vessel walls from elevated homocysteine results in accelerated plaque growth, followed by cholesterol and lipid deposition.159,159A Clinical and epidemiological research has shown that atherosclerosis throughout the body correlates with blood levels of homocysteine. Elevated homocysteine is a powerful and independent risk factor, as strong or stronger than other well-documented risk factors.150-152 Homocysteine is metabolized by enzymes that require metals and other dietary factors.160-169

Essential Free Radical Reactions

Life cannot exist without a balance of controlled free radical reactions. Life in the presence of oxygen requires antioxidant protection. Cellular energy production requires transfer of electrons across mitochondrial membranes within cells. For every electron that crosses the mitochondrial membrane, a superoxide radical is produced. Antioxidants prevent damage to vital intracellular structures during this process. Humans cannot utilize food for fuel without continuous cellular oxidation-reduction reactions, which produce free radicals as a byproduct. Oxygen is breathed in through the lungs and transported in the blood to every cell in the body, where oxidation reactions produce life-supporting energy. Red blood cells even produce free radicals during the binding and release of oxygen and carbon dioxide by hemoglobin.

Superoxide radicals are released during oxidative phosphorylation of ATP. Cellular protection from the superoxide radicals is provided by mitochondrial superoxide dismutase (SOD), a manganese-containing enzyme. The average American diet contains suboptimal amounts of manganese.170 SOD in the cytoplasm of cells requires both zinc and copper for activity. Those trace elements are also marginal to deficient in the average American diet.170 If the integrity of cell membranes is not protected by adequate SOD, then the activity of other vital enzymes contained within cell membranes is compromised.171

The metabolism of many chemicals, including most prescription drugs, artificial colorings and flavorings, petrochemicals, and inhaled fumes, takes place in the endoplasmic reticulum of liver cells and other organs. That detoxification process releases hydroxyl free radicals and peroxides.98, 98A,101,102,125-127 Glutathione peroxidase, vitamin C, and other antioxidants must be present in adequate supply to prevent chain reactions of damaging free radicals. In that way, drugs and other chemical exposure cause increased production of free radicals, which may then exceed the threshold of antioxidant protection.98, 98A,101,102 The resulting excess of free radicals may then ¬ multiply further, in a chain reaction, magnifying the damage by a million times or more.98, 98A,102

Synthesis of prostaglandins and leukotrienes from unsaturated fatty acids also results in the release of free radicals.102,106, 106A 128 Lacking sufficient antioxidant protection, prostaglandin production may become imbalanced. For example, production of thromboxane can increase, and prostacyclin can decrease in the presence of lipid peroxides.98, 98A,101,102 Thromboxane is associated with atherosclerosis while prostacyclin acts to prevent arterial plaque.

Leukocytes and macrophages normally produce free radicals. Disease-causing organisms are ingested and destroyed by free radicals during phagocytosis. The leukocytes use free radicals much like “bullets” against an invading army.172,173 Antioxidants localize and limit the damage caused by the free radicals. If antioxidant protection is inadequate, free radicals migrate into adjacent tissues and produce inflammation, manifested by redness, heat, pain, and swelling.

Without antioxidant enzymes, we would die very quickly. The aging process is accelerated by the age-related decrease in antioxidant protection.98, 98A,101,102 An extreme example of accelerated aging is the disease known as progeria, caused by hereditary absence of free-radical protective enzymes. Within ten to fifteen years after birth, a victim of that genetic mutation will experience every aspect of the aging process, including wrinkled, dried, and sagging skin, baldness, bent and frail body, arthritis, and advanced cardiovascular disease. Administration of an antioxidant has successfully slowed one form of progeria. Heredity determines each individual’s unique resistance to free radical mediated disease; therefore, there is a wide variation in tolerance to the dietary and lifestyle stresses that increase free radical production.

Oxygen Toxicity

The process of free radical pathology is often referred to as “oxidative stress.” Ground state or unexcited atmospheric oxygen has the unique property of being both a free radical generator and a free radical scavenger.98, 98A,102,174,175 Although a liter of normal atmospheric air on a sunny day contains over one billion hydroxyl free radicals,122 oxygen at normal physiologic concentrations in living tissues neutralizes more free radicals than it produces.176 When oxygen concentration falls below normal levels, as occurs with diseased arteries and ischemia, oxygen becomes a net contributor to free radical ¬ production.101,102,177,77A

Oxygen in high concentrations for prolonged periods of time can cause toxicity and even death, primarily by free radical damage to the lungs and brain. Under proper conditions, however, intermittent high-pressure oxygen administered for short periods in a hyperbaric chamber can stimulate an adaptive increase of intracellular superoxide dismutase, an enzymatic antioxidant.175,178 Too much or too little oxygen can be equally harmful. High levels of oxygen should therefore only be given in short pulsatile exposures, to stimulate an adaptive increase in antioxidant defenses without causing harm. It is now believed that the pulsatile nature of exposure is important for benefit.

Protection Against Oxygen Free Radicals

Normal oxygenation of tissues strengthens defense against free radicals. Aerobic exercise stimulates blood flow, improves oxygen utilization, and increases oxygenation of distal capillary beds. Increased tissue oxygenation during exercise thus acts to protect against free radicals and reduces free radical related disease.

Trace elements are essential components of antioxidant metallo-enzymes. Each molecule of mitochondrial SOD contains three atoms of manganese. Each molecule of cytoplasmic SOD contains two atoms of zinc and one atom of copper. Each molecule of glutathione peroxidase contains four atoms of selenium. Catalase and peroxidase contain iron. Elemental selenium is an antioxidant, independent of its function as an enzyme co-factor.102

The human body lacks an intrinsic defense against one destructive free radical precursor—excited state singlet oxygen. When superoxide radicals exceed a concentration that can be neutralized by SOD, they spontaneously convert to singlet oxygen. Polynuclear aromatic hydrocarbons and aldehydes found in tobacco tar and tobacco combustion products produce free radicals in the body, including singlet oxygen. The most important protection against singlet oxygen is dietary intake of beta-carotene; a precursor form of vitamin A.179-183 Epidemiological evidence correlates increased dietary beta carotene with reduced incidence of cancer. Fully active vitamin A lacks this protective activity.101,102

Beta-carotene is inactivated by free radicals and must be re-activated by other antioxidants, including vitamin C. Vitamin C is inactivated in the process. Cigarette smoke depletes vitamin C. If smokers are given beta-carotene, the oxidized beta-carotene further depletes vitamin C and other antioxidants. It is therefore important to supplement with a full spectrum of antioxidants and micronutrients simultaneously. In one study, an increase in lung cancer was reported when beta-carotene alone was given to smokers. If beta-carotene and vitamin C are not supplemented simultaneously, beta-carotene alone could worsen a preexisting deficiency of vitamin C. That would explain the seemingly paradoxical report of increased cancer with beta-carotene supplementation. Many different antioxidants work together in harmony. An antioxidant is inactivated when it encounters a free radical, and must in turn be reactivated by the next antioxidant in a cascade—a multi-step process.184-187 All antioxidants must be present simultaneously for optimal protection.

Vitamin E (tocopherol), vitamin C (ascorbate), beta-carotene, selenium in glutathione peroxidase, the amino acid cysteine in reduced glutathione, riboflavin, niacin, and a spectrum of other antioxidants are all interrelated in a recycling process that protects against free radicals. If all of those antioxidants are present in optimum amounts, they are continuously restored to their active forms, after becoming inactivated by free radicals.

The process proceeds as follows: vitamin E or beta-carotene neutralizes a free radical by becoming oxidized to tocopherol quinone or oxidized beta-carotene. Tocopherol quinone and oxidized beta-carotene are recycled into antioxidant vitamin E (tocopherol) or reduced beta-carotene by vitamin C, which is in turn oxidized to dehydroascorbate. Interestingly, the ratio of ascorbate to dehydroascorbate diminishes progressively with age and no species of animal survives when that ratio falls below one to one.102 This can be corrected by supplementation.

Inactive vitamin C (dehydroascorbate) is reduced to active vitamin C by glutathione peroxidase (which must contain selenium). Glutathione peroxidase is returned to its active form by oxidation of reduced glutathione. A vitamin B-2 (riboflavin)-dependent enzyme, glutathione reductase, returns oxidized glutathione to its active form. Glutathione reductase is reactivated by a vitamin B-3 (niacin)-dependent enzyme, NADH, which is oxidized to become NAD. NAD is metabolized in the electron transport system, passed from step to step down the carboxylic acid (Kreb’s) cycle; potentially destructive energy that originated as a free radical is redirected to useful metabolism. Subsequent steps in energy metabolism require every nutritional vitamin, mineral, and trace element.

This stairstep cascade of oxidation-reduction pathways demonstrates that each component depends on an adequate supp1y of all other components in the chain.124,124A A chain is only as strong as its weakest link. This interdependence explains the sometimes-equivocal results that are reported from clinical trials utilizing just one antioxidant. For years medical scientists have been conducting research by supplementing only vitamin E, beta-carotene, selenium, or vitamin C. Although results were often positive,180-182 benefits would have been much greater had a whole spectrum of essential nutrients been supplemented simultaneously.184-188 If free radical production in the body exceeds the neutralizing capacity of this system, serious damage to cell membranes, protein molecules, and nuclear material (DNA) results.98, 98A,101,102,111,119,121,123

A comprehensive understanding of free radical defenses provides a rationale for nutritional supplementation with a wide spectrum of multiple vitamins and trace elements, in safe amounts and in proper physiologic ratios. Although large amounts of water-soluble vitamins are rapidly excreted, transient elevations in tissues are achieved shortly following ingestion, which saturate those tissues and provide additional free radical protection and enhanced metabolic function.102

An 18-year nutritional study of thousands of people, published by researchers at the University of California, Los Angeles, showed that daily intake of a multiple vitamin-¬ mineral supplement that contained at least 500 mg of vitamin C could extend average life expectancy by up to six years.189

Increased Production of Free Radicals

When free radicals in living tissues exceed safe levels, the result is cell destruction, malignant mutation, tumor growth, damage to enzymes, and inflammation, all of which manifest clinically as age-related, chronic degenerative diseases. Each uncontrolled free radical has the potential to multiply by a million-fold in a chain reaction, much like a nuclear explosion.98, 98A,101,102,111,119,121,123

Dietary fats, especially polyunsaturated fats, are major sources of pathological free radicals. Double bonds on unsaturated fatty acids combine very readily with oxygen and can produce lipid peroxides. This occurs both after consumption and while exposed to the atmospheric oxygen prior to consumption.

Lipid peroxidation begins when fats and oils are exposed to air, and is greatly accelerated when heated. Oxidation of fats and oils is catalyzed and hence accelerated enormously by metallic ions, especially free iron and copper. For example, peanuts crushed to make peanut butter are rich in both iron and copper, which are freed into the unsaturated oil when the peanut is disrupted. Iron and copper are potent catalysts of lipid peroxidation and increase the rate of rancidity of peanut oil by up to a million times. Oxidized fats and oils are commonly called rancid; however, extensive peroxidation can exist in some oils without a detectable rancid odor or taste.102 Lipid peroxidation occurs during the manufacture of many foods and cooking oils.102

The more unsaturated the oil in fatty acids, the more readily peroxidation will occur. The rate of peroxidation is proportional to the square of the number of unsaturated bonds in each fatty acid molecule. Factors that further increase the rate of peroxidation include heat, oxygen, light, and trace amounts of unbound metallic elements.102,119 Oils prepared in the dark, at low temperatures, in an atmosphere of pure nitrogen, and with added fat-soluble antioxidants such as vitamin E, would be best for nutritional use.102 Such oils are not commercially feasible. The alternative is to eat oil-containing foods, such as nuts and seeds, in their natural state, without crushing until chewed and swallowed. Dietary supplementation with insurance doses of antioxidants can help to prevent free radical damage in the body, even when oxidized fats and oils are eaten.

Unsaturated vegetable oils often contain trace amounts of iron and copper and are routinely exposed to heat and oxygen when foods are fried. That creates the worst possible combination. Hence, the admonition to limit consumption of fried foods. Oils used in the manufacture of salad dressings, such as mayonnaise, often contain high concentrations of lipid peroxides. The poorest quality oils are commonly used to produce commercial food products because heavy seasoning masks rancidity.102

Peroxidation and hydrogenation of vegetable oils during the manufacture of margarine and shortening results in cis- to trans-isomerization. Trans-isomerization alters the three-dimensional configuration of dietary fatty acid molecules from their normal ” coils to straightened “trans” configurations. Trans-fatty acids are then incorporated into cell membranes in the place of naturally occurring cis forms, weakening the membrane structure and impairing function of phospholipid-dependent enzymes imbedded in cell membranes.101,102,218,219 Substrate recognition by enzymes that synthesize cell membranes is not adequate to distinguish between these two forms.102

Phospholipids that compose cell membranes are easily damaged by free radicals, as already explained. Dietary intake of peroxidized fats initiates that process. The prostaglandin precursors—arachidonic and linoleic acids—are depleted in the process, as measured by gas chromatography.101,102 Cell membranes containing trans-fatty acids have impaired fluidity and increased permeability that interfere with transport of sodium, potassium, calcium, magnesium, and other substances. Receptors for insulin and other hormones are disturbed.98,101,102 Damage from trans-isomerization of fatty acids is cumulative with lipid peroxidation.98, 98A,101,102

Very little attention is commonly paid to the quality of dietary fats and oils. Emphasis has mistakenly been placed on the ratio of saturated to unsaturated fatty acids, irrespective of lipid peroxidation and trans-isomerization. Contrary to conventional wisdom, unsaturated fats are more toxic than saturated fats.102 Margarine contains far more peroxides and trans-fatty acids than butter. Moderating consumption of all fats and oils is desirable.102 It has been shown epidemiologically that margarine consumption is a risk factor for heart disease, contrary to advertisements for preventive benefit.192

The quantity and quality of dietary fat are as important or more so than the ratio of unsaturated to saturated fatty acids.98, 98A,101,102 If dietary fats and oils are obtained from fresh, whole, unfractionated, and unprocessed foods, they will be minimally oxidized and will produce healthy cell membranes with normal cis-fatty acid configurations. They will preserve a normal balance of prostaglandins. Although fully saturated fats are not as easily oxidized, all animal fats contain some unsaturated fatty acids and cholesterol, both of which are subject to oxidation. Animal experiments have shown that as little as one percent of dietary cholesterol consumed in oxidized form can contribute to atherosclerosis. Supplemental antioxidants reduce that risk.144,145,193

How much dietary fat and oil can one tolerate without risk? Evidence indicates that between 25 to 35 percent of dietary calories as fat can be both safe and nutritious, if attention is paid to the quality and source of the fat, as described above, and if supplemental vitamins, minerals, and trace elements are used.101,102 The more oxidized the fats, the less well they are tolerated. In the United States, an average of 45 percent of dietary calories are consumed as fat, mostly of poor quality, with no consideration for rancidity or cis- to trans-isomerization. Half the population takes little or nothing in the way of nutritional supplements. Lipid peroxidation occurs much more slowly when foods are frozen.102

Free radical damage contributes to senility, dementia, and other nervous system diseases, including Alzheimer’s and Parkinson’s syndromes. The brain and spinal cord contain the highest concentration of fats of any organ. Nervous system fats are also very rich in highly unsaturated arachadonic and docosahexanoic acids. Because the rate of lipid peroxidation increases exponentially with the number of unsaturated carbon-carbon double bonds, docosahexanoic and arachadonic acids peroxidize many times more readily than most other lipids. The brain and spinal cord therefore require added antioxidant protection.

To provide that extra protection, vitamin C is concentrated in the brain by metabolically active pumps in the blood-brain barrier. Ascorbate is 100 times more concentrated in the brain and spinal cord than in other organs.102 Two ascorbate pumps operate in series. The first increases the concentration ten-fold from blood to cerebrospinal fluid. A second pump concentrates vitamin C by another factor of ten between cerebrospinal fluid and the subdural space. The ¬ disappearance rate of vitamin C from spinal fluid can be used to indicate the extent of damage and subsequent lipid peroxidation following ischemia or trauma to the central nervous system.194,195

Experimental spinal cord injuries in animals have been used to show benefit from treatments based on free radical protection. A minor contusion to the spinal cord results in rapid breakdown of unsaturated fatty acid sheaths surrounding nerve pathways. Following a contusion, capillaries leak blood. Erythrocytes hemolyze, releasing free iron and copper. Those metals are potent catalysts and react with oxygen to massively increase the rate of lipid peroxidation.102 The chain reaction of oxidative damage and inflammation that occurs has been extinguished experimentally in two ways in animals: 1) The spinal cord has been exposed and irrigated with a potent free radical scavenger, such as dimethyl sulfoxide (DMSO); and 2) iron and copper catalysts have been inactivated by bathing the injured area in a chelating solution containing EDTA.102,109,140,191,196-198

Intravenous DMSO in large doses has been reported to prevent paralysis and permanent damage in victims of spinal cord and brain injuries. Results thus far indicate that if treatment is begun within the first thirty minutes, or at most within the first two hours, the outcome is much better than would otherwise have been expected.199-214

Chelation therapy with EDTA combined with dietary fat restriction has been reported to alleviate or temporarily reverse the progression of multiple sclerosis (MS).34,215,216 MS victims experience degeneration of the fatty myelin sheaths that insulate nerve pathways in the brain and spinal cord. Although lipid peroxidation may be only one link in the chain of cause and effect, it is sometimes possible to slow this devastating disease by using treatment principles that reduce free radical pathology.

Tobacco and Alcohol

Habitual tobacco use and excessive alcohol use lead to disease and premature death. Alcohol causes damage and scarring to the liver and cancer in the mouth and digestive organs.217 Alcohol is metabolized to acetaldehyde, which is a potent free radical precursor. Acetaldehyde is closely related to formaldehyde (embalming fluid), and causes cross-linkages of connective tissue by free radical reactions (like tanning leather).101,102,218,219 Alcoholic cirrhosis of the liver might therefore be considered quite literally a form of pre-death ¬ embalming.

Tobacco smoke contains polynuclear aromatic hydrocarbons, which are potent precursors of free radicals. Those substances can easily overwhelm the body’s free radical defenses and speed the onset of cancer, atherosclerosis, and other age-related degenerative diseases.220 Processed tobacco also contains cadmium, a heavy metal ten times more toxic than lead, which acts as a catalyst of free radical reactions and displaces zinc in metallo-activated enzymes.

Cell Membrane Metabolism

Every cell in the body is enveloped in fat in the form of bipolar, phospholipid membranes. Spanning those membranes are large proteins, enzymes, molecular pumps, and receptors for various hormones and peptides. Cell membranes are metabolically very active and have the characteristics of a viscous fluid. They are constantly changing. Cell membranes have one-way permeability to substances that must be kept out of or inside of cells. The water-soluble, polar ends of phospholipid molecules line up on the inner and outer surfaces of the cell membrane, bathed in aqueous fluids. The fat-soluble, nonpolar tails point toward the center of the membrane, intertwining with the fatty tails of similar molecules extending from the opposite surface, traversing the interior of the cell-wall membrane. The normal, curly cis-configuration of membrane phospholipids allows them to twist around each other and grasp cell-wall proteins, enzymes, and other constituents within the membrane, holding them in proper position. The cis-curvature of healthy lipids is essential to integrity and metabolic activity of cell membranes.98, 98A,101,102

Unoxidized cholesterol is widely disbursed within cell membranes and acts as an antioxidant.102 Oxidized dietary cholesterol produced in food processing offers no such protection and is atherogenic.102,144 When free radicals occur in the vicinity of a cell, unoxidized cholesterol, vitamin C, vitamin E, coenzyme Q-10, and the entire array of antioxidant defenses are needed to prevent damage.

Large enzymatic proteins span the full thickness of cell membranes and act as metabolically active “pumps.” They are bathed in plasma on the exterior and extend into the cytoplasm within the cell. One such pump keeps sodium ions out of the cell and potassium within, against a powerful diffusion gradient. Another keeps calcium out and magnesium in. Cellular organelles, including mitochondria, lysosomes, endoplasmic reticuli, Golgi bodies, and nuclear DNA are enveloped in similar bipolar lipid membranes that also contain many energy-dependent transport mechanisms. Mitochondrial membranes are protected by coenzyme Q-10, an antioxidant necessary for safe energy production. Mitochondria are the power plants of cells, and they continually produce free radicals during transport of electrons, in the process of oxidative phosphorylation. Damage to mitochondria causes premature aging. A spectrum of antioxidants is necessary to prevent those free radicals from destroying mitochondria.98, 98A,101,102

Receptors on cell membranes for neurotransmitters, insulin, hundreds of different oligopeptide regulators, and hormones are also subject to damage by free radicals. The ¬ calcium-magnesium and sodium-potassium pumps become weakened with advancing age, allowing harmful levels of calcium and sodium to enter cells. Free radicals damage nuclear membranes, altering nuclear pores and chromosomes, and causing mutations with resulting impairment of protein synthesis and cell replication. Free radical mutations of DNA can cause uncontrolled cell division and cancer.

Free radicals increase the activity of guanylate cyclase, which can stimulate uncontrolled cell multiplication. Lymphoid tissues are very rich in unsaturated fatty acids, and free radical damage can easily cause immunologic abnormalities.102 The immune system may subsequently attack the body’s own tissues in so-called autoimmune diseases or it may weaken and fail to recognize and destroy disease-causing organisms and malignant cells.98, 98A,101,102,106

Calcium Metabolism

Free radical damage to the calcium-magnesium pump allows excessive calcium to diffuse into the cell. Calcium is 10,000 times more concentrated outside than inside of cells. The calcium pump must constantly work against this high diffusion gradient. The reverse is true of magnesium. If the pump cannot prevent calcium from leaking into cells, and keep magnesium from leaking out, the cell is poisoned and soon dies.

Calcium activates phospholipase-A2, which cleaves arachadonic acid from membrane phospholipids. Increased levels of arachadonic acid can in turn create an imbalance of prostaglandins and leukotrienes, creating more free radicals in the process.101,102 Leukotrienes are potent mediators of inflammation and attract leukocytes. Leukocytes, as noted previously, release superoxide free radicals during phagocytosis. Leukocytes, stimulated by leukotrienes, can overpower local antioxidant defenses, leading to inappropriate inflammation and damage to surrounding tissues.101,102,227 Small capillaries and arterioles dilate, causing edema and leakage of erythrocytes through blood vessel walls. Platelets produce microthrombi. Erythrocytes hemolyze, releasing free copper and iron, which catalyze progressive oxidative damage to adjacent tissues. This results in an inflammatory chain reaction, beyond normal control mechanisms.

Free radical damage causes calcium to leak into smooth muscle cells in arterial walls. Calcium binds to calmodulin, activating myosin kinase, which in turn phosphorylates myosin. Myosin and actin constrict, causing the muscle cells to shorten. By this mechanism, excess calcium within arterial smooth muscle cells causes spasm. The same occurs in cells of the myocardium. When muscle fibers encircling arteries constrict, blood flow is reduced. Calcium channel blockers relieve symptoms by slowing abnormal entry of calcium into cells, but they do not correct the underlying cause of the problem—free radical disruption of cell membranes.101,102,222

Myocardial cells are weakened by excessive intracellular calcium, lowering the efficiency of oxygen utilization and placing an extra burden on an already impaired coronary artery system. If a coronary (or other) artery is partially occluded by atherosclerotic plaque, a small amount of spasm superimposed on a preexisting partial blockage can easily cause ischemia. A myocardial infarction can result from spasm alone, even in a coronary artery completely free from plaque.223 Thromboxane and serotonin are released by platelets in the presence of free radicals.101,102 Thromboxane and serotonin also cause arterial spasm.

Excessive intracellular calcium can result from a variety of other factors. Ionized plasma calcium, the metabolically active fraction not bound to protein, slowly increases with age. The higher the concentration of ionized calcium outside a cell, the harder the calcium pump must work to prevent excessive calcium from leaking in. Naturally occurring calcium channel antagonists can slow the calcium influx. These include dietary magnesium, manganese, and potassium. Magnesium and manganese intakes are suboptimal in the average American diet.170 Diets are rarely deficient in potassium, but an excessively high ratio of dietary sodium to potassium is common, allowing excessive sodium to diffuse into cells, further weakening metabolism. Losses of potassium and magnesium from diuretic therapy can potentiate this problem.

The efficiency of energy metabolism can be impaired by stress. Stress increases circulating catecholamines and inhibits production of ATPase. The calcium and sodium pumps both require ATPase. Cells lose potassium and magnesium and retain calcium and sodium at a greater rate under stress because of relative inhibition of both magnesium-calcium ATPase and sodium-potassium ATPase.

Catecholamines produce free radicals when they are metabolized.101,102 If free radicals in the central nervous system exceed defenses, stress-related catecholamines cause free radical damage to neuron receptors. That is a partial explanation for stress-related nervous disorders. Breakdown products of dopamine, a catecholamine neurotransmitter, can also cause free radical damage to neuronal receptor sites in the brain. That is hypothesized to be one factor in Parkinson’s syndrome and some types of schizophrenia.224 Neuronal receptors for norepinephrine can also be damaged by free radicals, leading to depression. Heart disease has also been shown to result from catecholamine-induced free radicals.225

In recent animal experiments, primates subjected to stress were found to have an increased incidence of atherosclerosis, even while fed a diet that is otherwise protective. Increased catecholamine metabolism provides an explanation.

Dementia of the Alzheimer’s type is thought by some authorities to be caused by brain cell destruction from free radicals.204 Arrest or improvement in that condition has been reported following treatment with deferoxamine, an iron chelating agent.226 EDTA also removes iron and aluminum very effectively. Free, unbound iron is a potent catalyst of lipid peroxidation. Accumulations of aluminum, combined with lipid and protein breakdown pigments are found in brains affected by both Parkinson’s and Alzheimer’s, although it is not known whether those are late events and not causative factors—much like the accumulation of calcium and cholesterol in arterial plaque.

Before the significance of free radical pathology was recognized, it was hypothesized that EDTA chelation therapy had its major beneficial effect on calcium metabolism. It now seems more probable that calcium is just another link in the chain of cause and effect created by free radical damage. EDTA can influence calcium metabolism in many ways, but direct action on calcium is not adequate to explain the many benefits that follow chelation.

EDTA lowers ionized plasma calcium during infusion. The body then attempts to maintain a homeostasis by producing parathormone.227 The intermittent three- to four-hour pulses of increased parathormone caused by EDTA can have a measurable effect on bone metabolism.228 Frost’s concept of bone metabolism, known as the Basic Multicellular Unit (BMU) theory, is accepted by other experts.229 The BMU theory helps to explain the causes and treatment of osteoporosis and osteopenia.

The BMU is a group of metabolically active cells that control the turnover of approximately 0.1 cubic millimeter of bone tissue. When a BMU is activated, it goes through a cycle consisting of an initial three to four weeks of bone absorption (osteoclastic phase) followed by a two- to three-month period of bone reformation (osteoblastic phase). Net increase or decrease in bone density at the end of the entire three- to four-month cycle depends on the rate and completeness of bone turnover. Hormone regulation of BMUs also includes calcitonin, growth hormone, thyroxin, and adrenal corticosteroids, but parathormone remains the most important controlling factor.228

Chronically high levels of parathormone have long been known to cause net bone destruction; but brief pulsatile increases in parathormone, as occur during intravenous EDTA chelation therapy, cause an increase of new bone formation.52,230

Anabolic activation of BMUs by pulsatile parathormone secretions provides one possible hypothesis for delayed benefit seen in chelation patients, although the theoretical explanations in this paper are more likely to be important. It is theorized that calcium deposits might be removed from arteries and other soft tissues for utilization in new bone formation.

Note that effect of EDTA on calcium factors do not occur when calcium EDTA is infused. Only disodium EDTA can bring these potential benefits.

calcium metabolism

In their original studies, Meltzer and Kitchell used EDTA to treat ten men who were severely disabled by heart disease and suffered from intractable angina. After approximately twenty infusions of EDTA, therapy was discontinued because of initially disappointing results. Three months later, nine out of ten patients returned to report marked relief of angina, despite no change in their lifestyles, such as altering smoking or nutritional habits.7 This three-month delay in achieving full benefits has remained a consistent observation by chelating physicians over the years. The delay in achieving full benefit suggests that EDTA activates some type of long-acting healing mechanism.

As an aside, postmenopausal women who are not supplemented with estrogen experience a large increase in follicular stimulating hormone (FSH). Elevated FSH interferes with new bone formation by BMU cells and is regarded as a contributing factor in postmenopausal osteoporosis.231

Before free radical pathology was discovered, it was hypothesized that removal of calcium from atherosclerotic plaque and from pathological cross-linkages could explain most of the benefits seen from chelation. Correction of molecular cross-linking may also be an important benefit from EDTA. Undesirable cross-linkages include disulfide bonds caused by free radical reactions that increase with age, and from intermolecular bridging by divalent cations that increase with age, such as calcium, lead, cadmium, aluminum, and other metals. EDTA can remove those metals. Chelation can also reduce abnormal disulfide cross-linkages. Reduction of cross-linking between molecules acts to restore the elasticity of vascular walls and other tissues that is lost with age .36,37

Improvements in blood flow may not be detectable on arteriograms, despite marked clinical improvement. Research shows that arteriograms are limited, and can only measure the diameter of an artery to within plus or minus 20 percent (as described in the introduction). Although counterintuitive, it is true that with perfect laminar blood flow, a mere 19 percent increase in the diameter of an artery doubles flow of blood. In a plaque-filled vessel with turbulent flow, less than 10 percent increase in diameter will result in a doubling of blood flow. This can be proved using Poiseuille’s law of hemodynamics, as explained in textbooks of medial physiology.

In an organ with compromised circulation, a 25 percent increase in blood flow could bring significant functional improvement and relief of symptoms. Changes in diameter of such small magnitude cannot be detected on either arteriograms or ultrasound imaging.

Lipid Peroxidation

EDTA can greatly reduce the excessive production of free radicals.102,143,232  Traces of unbound metallic ions act as catalysts of uncontrolled proliferation of free radicals in tissues. EDTA binds those ionic metals, making them chemically inert and rapidly removing them from the body. The amount of metal ions necessary to catalyze lipid peroxidation is so minuscule that the tiny traces remaining in distilled water can initiate and accelerate those reactions.102,176 Metals incorporated into metallic enzymes are tightly bound and not readily accessible to EDTA. Some essential elements are briefly removed, however, and may require oral supplementation for replacement.

To catalyze lipid peroxidation, a metallic ion must easily change electrical valence by one unit. Two essential nutritional elements, iron and copper, are potentially the most potent catalysts of lipid peroxidation, although copper is often deficient, and not prevalent enough in the body to be clinically important in that regard. With age, catalytic iron accumulates adjacent to phospholipid cell membranes, in joint fluid, and in cerebrospinal fluid. Iron is released into tissues following trauma and ischemia. This unbound form of extracellular iron causes free radical tissue damage, evidenced clinically as inflammation.110,119,121,233-238

Toxic Heavy Metals

EDTA has long been an accepted treatment for acute heavy metal poisoning. Toxic heavy metals can impede metabolism in a variety of ways. Heavy metal toxicity is rare these days because they have largely been eliminated from the environment. In the absence of ongoing exposure, the body slowly excretes metallic toxins, making chelation therapy unnecessary.  A blood test is the easiest and most accurate way to measure for this problem. Poisonous metals in excess, such as lead, mercury, and cadmium, react avidly with sulfur-containing amino acids on protein molecules. When lead reacts with sulfur on the cysteine or methionine moiety of an enzyme, enzyme activity is reduced or destroyed. Lead also displaces zinc in zinc-dependent enzymes. Chelation therapy reactivates enzymes by removing such toxic metals. The average concentration of lead in human bones has increased since the Industrial Revolution.239 Bone lead is in equilibrium with other vital organs and is released into the circulation with a fever and under stress, increasing toxicity when it can be least tolerated.240

Lead destroys the antioxidant properties of glutathione and glutathione peroxidase. Lead reacts vigorously with sulfur-containing glutathione and prevents it from neutralizing free radicals. As previously described, reduced glutathione is an essential antioxidant in the recycling of vitamins E and C, glutathione peroxidase, glutathione reductase, and NADH. Lead therefore poisons free radical protective activity of the entire cascade of antioxidant protection.

Lead reacts with selenium even more avidly than sulfur, inactivating the selenium-containing enzyme, glutathione peroxidase. In addition to performing its role in the antioxidant recycling system, that enzyme protects directly against lipid peroxides. Other toxic heavy metals also inactivate glutathione peroxidase. Testing for levels of those metals, as well as clinical evaluation for adequacy of trace element nutriture, is important in the initial evaluation of a patient prior to chelation therapy.170,241-266

The wide variety of benefits reported following EDTA chelation therapy can be understood using the above concepts. EDTA cannot easily chelate metallic ions when they are tightly bound within metal-containing enzymes and metal-binding proteins. Most EDTA remains extracellular.  On the other hand, when metals accumulate in unbound form, able to act as catalysts of uncontrolled lipid peroxidation, EDTA can easily bind and remove them. Iron accumulates with age at abnormal locations, where it accelerates free radical damage.34,233-238, 238A EDTA binds much more tightly to iron and other potential free radical catalysts than it does to calcium and toxic heavy metals. EDTA will only bind calcium if those other ions are not present.34

Excretion of metals after EDTA

Iron accumulates more slowly in women during the childbearing years because of monthly menstrual blood losses. Because of that monthly iron loss, younger women have significant protection against atherosclerosis. That protection is lost at menopause. Body iron stores, best reflected by a combination of serum ferritin and transferrin saturation, accumulate in men four times more rapidly than in premenopausal women.267,267A Chronic inflammation anywhere in the body can falsely elevate serum ferritin levels and transferrin saturation is needed to confirm the meaning of ferritin levels.]The risk of atherosclerosis is  four times greater in men in this same age group. Data from the Framingham study show that two years after a hysterectomy, a woman’s risk for cardiovascular disease becomes equal to a man’s, with or without hormone replacement. This occurs even if the ovaries are not removed.268-270 Those observations indicate that slower iron accumulation is primarily responsible for reduced atherosclerosis in premenopausal women, although estrogen also has antioxidant benefit.271 Periodic donation of blood to the blood bank significantly prolongs life expectancy.272 The fact that iron is a potent catalyst of lipid peroxidation provides a link between these clinical and epidemiologic findings. EDTA has a very high affinity for unbound iron and rapidly removes it from the body.

The affinity of EDTA to bind various metals at physiologic pH, in order of decreasing stability, is listed below. In the presence of a more tightly bound metal, EDTA releases metals lower in the series and binds to the metal for which it has a greater affinity.273 Calcium is near the bottom of the list, while iron and some toxic metals are near the top.

Chromium 2+

Iron 3+

Mercury 2+

Copper 2+

Lead 2+

Zinc 2+

Cadmium 2+

Cobalt 2+

Aluminum 3+

Iron 2+

Manganese 2+

Calcium 2+

Magnesium 2+

In clinical practice, chromium, mercury, and copper are not removed in any significant amount by EDTA, indicating that they are already more tightly bound than is possible by EDTA.273A

Magnesium is a calcium antagonist and is relatively deficient in many chelation patients. Magnesium is the metallic ion least likely to be removed by EDTA and is often given as an oral supplement.

Lasting inhibition of excessive free radicals by EDTA offers an explanation for the data from Switzerland, which documented a 90 percent reduction in deaths from cancer in a large group of patients who were chelated and then carefully followed over an eighteen-year period. Chelation patients were compared with a statistically matched control group. Death rate from cancer was ten times greater in the untreated group, compared to the death rate of patients who had been previously treated with EDTA (P=0.002).38 Cardiovascular death were unfortunately not analyzed. One common denominator of both cancer and atherosclerosis is free radical oxidative damage to molecules.98, 98A,101,102 Calcium-EDTA was administered in that study, which precludes any direct effect on calcium metabolism as an explanation for outcomes. Removal of free radical catalysts seems the likely explanation. Demopoulos first proposed that chelation be used to control free radical pathology.102,121 He also pointed out that many antioxidants have chelating properties.98, 98A,

EDTA increases the efficiency of mitochondrial oxidative phosphorylation and improves myocardial function, quite independently of any effect on arterial blood flow.274 Treatment with deferoxamine, an iron chelator, has been shown to improve cardiac function in patients with increased iron stores.275 In addition, removal of iron with deferoxamine reduces inflammatory responses in animal experiments.276 Sullivan has suggested that periodic donation of blood be studied as a way to reduce the risk of atherosclerosis in men and postmenopausal women.271-272

By reducing damage from free radicals, EDTA chelation therapy can support normal healing. The time required for healing of damaged tissues gives us another explanation for the time lapse of several months following chelation before full benefit is achieved. By correcting the underlying cause of the disease process, and allowing time for subsequent healing, treatment with EDTA seems far superior to the mere suppression of symptoms achieved with so many other therapies.

Chelation and Atherosclerosis

Minor injuries and tears in blood vessel walls occur frequently in an ongoing way. Rather than heal, some cause scars or progress to further injury.

If an injury results in bleeding, homeostatic mechanisms quickly stop the flow of blood to prevent hemorrhage and death. This regulation of blood loss is under the control of a complex array of mechanisms, including hormones, prostaglandins, fibrin, and thromboplastin. Prostaglandins are produced and degraded continuously and very rapidly in endothelial cells and platelets. Prostaglandins have a half-life measured in seconds and must be constantly synthesized at a controlled rate and with a proper ratio between their various subtypes to maintain normal blood flow.

The two most important prostaglandins in that regard are prostacyclin and thromboxane. Prostacyclin reduces the adhesiveness of platelets, facilitating free flow of blood cells and plasma, and reducing the tendency for fibrin deposition and thrombus formation. Prostacyclin relaxes encircling muscle fibers in artery walls, reducing spasm. Thromboxane does the opposite. It causes intense spasm in blood vessel walls and stimulates platelets to adhere.277 In oversimplified terms, thromboxane may be considered as undesirable and prostacyclin as desirable. In actual fact, a proper balance must be maintained to protect against injury and hemorrhage, on the one hand, and to maintain normal circulation, on the other.

Synthesis of prostacyclin is greatly inhibited by lipid peroxides and free radicals, while thromboxane production remains unaffected. If lipid peroxides are present, either from dietary intake of peroxidized fats and oils or from nearby peroxidation of lipid cell membranes, less prostacyclin is produced to balance the effects of thromboxane.101,102,278

Ongoing damage to vascular endothelium occurs continuously in response to hemodynamic stresses. Damage may also be caused by disordered immunity or bacteria. With good health, minor vascular injuries are rapidly healed, initiated by a layer of platelets that coat the disrupted surface with a protective blanket.145 If free radical protection is inadequate and local controls have been taxed, the local increase of free radicals blocks the production of prostacyclin. Without prostacyclin, thromboxane is unopposed and causes the injured area of the arterial wall to attract platelets abnormally. This causes platelets to increasingly stick to each other. Platelets thus aggregate, and the growing layer of platelets traps leukocytes, which in turn produce more free radicals. A network of fibrin and microthrombi is formed, and erythrocytes become trapped. Some of the erythrocytes hemolyze, causing iron to be released. Catalytic iron then produces an explosive increase in free radical oxidation, oxidizing any cholesterol present and damaging phospholipids in cell membranes. Prostacyclin production continues to be inhibited for some distance along the blood vessel.

The resulting high concentrations of free radicals can damage nuclear material in arterial cells, causing mutation and uncontrolled cell replication. Lipid peroxides increase the activity of guanylate cyclase, which speeds mitosis.98, 98A,101,102 Platelets release growth factors. This sequence of events can produce an atheroma, an enlarging tumor consisting of mutated, rapidly multiplying, multipotential cells that have lost their high degree of differentiation and specialized function. Atheroma cells produce substantial amounts of connective tissue, collagen, and elastin. They also act as macrophages, ingesting cellular debris and oxidized cholesterol. The monoclonal theory of atheroma formation first proposed by Benditt279 most accurately fits these known facts. Cholesterol is oxidized by free radical activity, and some of the cholesterol oxidation products ingested by atheroma cells have a toxic form of vitamin D activity.144

We previously explained how intracellular calcium increases abnormally because of free radical damage to homeostatic mechanisms, especially in cell wall transport mechanisms. In its role as an antioxidant, unoxidized cholesterol acts to protect against free radical damage but becomes oxidized in the process. Some cholesterol is synthesized within atheroma cells under this stimulus.280 Oxidized cholesterol and cholesterol esters can thus accumulate within plaque. As a plaque gradually expands to exceed its blood supply it may ulcerate. A central core of the plaque eventually degenerates into an amorphous fibro-fatty mass containing varying amounts of calcium, cholesterol, connective tissue, and cellular debris. When this necrotic core ruptures, it releases embolic showers of plaque debris, causing further aggregation of platelets. Platelets release high concentrations of thromboxane and serotonin, lwhich may promote arterial spasm and ischemia.

Symptomatic ischemia usually does not occur until a blood vessel becomes 75 percent occluded. A meal laden with peroxidized fats will cause a sudden free radical insult, triggering an abrupt spasm or acute thrombosis, superimposed on a partial occlusion, resulting in an acute infarction.

Cell damage may occur in any part of the body. Cells swell and die as membranes become leaky and damaged. Membrane pump mechanisms become uncoupled or disabled. DNA damage results in mutations, atheromata, and cancer.98,98A,101,102 Lymphoid tissues and other cells of the immune system become damaged.98, 98A,101,102 Tissues become lose flexibility as cross-linkages occur in connective tissue, elastin, and protein molecules.

Organ functions deteriorate. Joints become hypertrophic, inflamed, and deformed with arthritis. Leukotriene production and prostaglandin imbalances accelerate inflammatory change in joints and other organs. Lysosomes rupture, releasing autodigestive proteolytic enzymes that further devastate cell contents. Lysosomes have been called the cells’ internal digestive organs and, when disrupted, cause cellular autodigestion. Free radicals inactivate selenium, creating inert selenium compounds and causing metabolic selenium deficiency. Cancer patients excrete selenium in amounts up to five times the normal rate, just when they need it the most.170

Antibody production and cellular immunity are also impaired by free radicals. Cells of the immune system are especially rich in unsaturated fats and are therefore more vulnerable to free radical oxidative damage. Oxidized cholesterol and lipid peroxides are potent immunosuppressants.101,102,106, 106A 144 Antigenic substances and malignant cells, which would otherwise be neutralized, can overwhelm a weakened immune system. Intact food molecules can leak across the gut wall undigested and are poorly tolerated.281-283 Adverse reactions to specific foods (“food allergies”) then appear. Normal free radical reactions in macrophages during phagocytosis of antigens become out of control, increasing inflammation. Adverse reactions are triggered by a variety of nutritious foods and environmental exposures to which the immune system becomes sensitized. This is an increasingly common cause of symptoms. Avoidance of sensitizing foods and other trigger factors becomes necessary to control symptoms.284-288

Antigenic properties and toxins released by Candida albicans, a yeast normally present in the body in small numbers, can increase and overwhelm the immune system.289-295 A struggling immune system may become over-reactive in other areas, attacking healthy tissues, leading to so-called autoimmune syndromes.

Atheroma and Cancer: Both Are Tumors

The development of cancer can take decades from the initiating event to the onset of symptoms. If cancer-promoting factors are removed, free radical damage can be repaired and healing can be aided by a more nutritious diet, antioxidant supplementation, and lifestyle improvements. In the early stages, malignant cells have the ability to transform back into a normal, benign state. For example, smokers who stop the use of tobacco have approximately the same risk of cancer ten years later as those who never smoked.102

Atherosclerotic plaque is actually a benign tumor, an atheroma, somewhat analogous to cancer. It does not metastasize but expands locally to occlude the flow of blood, or outgrow its blood supply and rupture. Plaque can regress with time, if causative factors are reduced. Free radical pathology is the common denominator for both atherosclerosis and cancer.

Treatment and Prevention of Diseases of Aging

(1) Diet

Oxidized fats and oils are best limited.101,102 Consumption of fats and lipids that have been processed, exposed to air, heated, hydrogenated, or otherwise altered should be avoided when practical. Consumption of refined carbohydrates that are depleted of trace elements (white flour, white rice, and sugar) should be minimized. Total caloric intake should be moderated to maintain weight within 20 percent of ideal body weight. The use of excessive salt should be avoided. Diets should contain ample amounts of fiber-rich whole grains and fresh vegetables. Patients suffering with chronic debilitating diseases must be stricter with diet. Clinical improvement involves a healing process, often requiring months or years to complete.

(2) Nutritional Supplements

A scientifically balanced regimen of supplemental nutrients reinforces endogenous antioxidant defenses. It is often not possible to receive optimal quantities of those substances from food alone.170 Trace elements can be toxic if taken to excess, and iron supplementation in the absence of deficiency will speed free radical damage. Iron should be supplemented only to treat deficiency states, confirmed by low serum ferritin and confirmed by low transferrin saturation.267A,271,296 Trace element supplementation should be under the supervision of a health care professional knowledgeable in nutrition. Dietary histories and biochemical may indicate the need for and type of supplementation.170,241-266,296,297

(3) Modification of Health-Destroying Habits

Tobacco: It is best to eliminate the use of tobacco altogether, but, if that is not possible, a marked reduction in exposure would be helpful. This applies to cigarettes, pipe tobacco, cigars, snuff, and chewing tobacco. Tobacco causes problems, even without combustion. Free radical precursors are absorbed from tobacco through the lining of the mouth and nose, even without inhaling smoke. A relatively healthy adult with supplemental intake of antioxidants may tolerate a small exposure to tobacco without an increased risk of cancer, but even a small amount may increase the risk of atherosclerosis.298

Alcohol: Many victims of degenerative diseases discover for themselves that alcohol is not well tolerated. For individuals with chronic illness, complete avoidance is advisable. A healthy adult should be able to tolerate and detoxify one to two ounces of pure ethanol per twenty-four hours (up to four eight-ounce glasses of beer, four small glasses of wine, or two shot glasses of hard liquor at most). That amount may be consumed in twenty-four hours without exceeding a healthy person’s capacity to metabolize alcohol and neutralize the resulting free radicals.101,102 But with daily use, tolerance may be lost.

(4) Physical Exercise

Moderate physical exercise, even a brisk walk several times per week will improve efficient utilization of oxygen. More vigorous aerobic exercise results in proportionately greater benefits. Lactate accumulates up to twice normal levels in tissues during endurance exercise.299 Lactate has chelating properties, and it is possible that some of the benefits of exercise may result from chelating effects of lactate in tissues during aerobic exercise.37

(5) EDTA Chelation Therapy

The use of EDTA to alter the balance and distribution of essential metallic elements, while at the same time removing excessive metals and free radical catalysts, has been shown to slow or arrest progression of diseases of aging. Other benefits of chelation can occur from uncoupling of disulfide and metallic cross-linkages between molecules, by normalization of calcium metabolism, by reactivation of enzymes poisoned by toxic metals, and by restoration of normal prostacyclin production along blood vessel walls. Lasting benefits follow a series of intravenous EDTA infusions, plus indicated nutritional supplementation and lifestyle improvements.

This well-documented, safe, and effective therapy deserves widespread recognition and acceptance.

NEWER RESEARCH

We still do not know for certain the most important ways in which how EDTA chelation therapy benefits atherosclerosis and other age-related diseases. We only know that it binds to metallic ions in the body. EDTA rapidly removes ionic metals via urinary excretion, and in the process it redistributes metals within in the body. We have many theories attempting to explain how EDTA reverses symptoms, improves cardiovascular function, enhances quality of life, and improves blood flow, but we still do not know the most important mechanism(s) of action.

A recent study by Frustaci and associates in Italy showed that nutritional trace elements accumulate to potentially toxic levels in diseased myocardium.300 Marked increases in intracellular concentrations of myocardial trace and elements were measured by neutron activation analysis in a control group of patients, compared with with coronary ischemia, valvular disease and idiopathic cardiomyopathy. The nutritional elements, iron, zinc, chromium and cobalt increased from three- to seven-fold in compromised myocardium. All metals become toxic at those levels.

Frustaci data

Table data above excerpted from:  Frustaci A, Magnavita N, Chimenti C, Caldarulo M, Sabbioni E, Pietra R, Cellini C, Possati GF, Maseri A. Marked elevation of myocardial trace elements in idiopathic dilated cardiomyopathy compared with secondary cardiac dysfunction. J Am Coll Cardiol. 1999 May;33(6):1578-83.

In patients with advanced valvular disease and idiopathic cardiomyopathy, intracellular metals increased to a similar extent as in those with ischemic coronary artery disease.

In another study, copper and zinc accumulations in the brain are implicated in Alzheimer’s syndrome. Alzheimer's researchers at the Massachusetts General Hospital have reported that buildup of copper and zinc in the brain causes the type of protein deposits that are a hallmark of Alzheimer's disease.301 Using mice bred to develop a form of Alzheimer's, they found that a metal chelator, clioquinol, neutralized those metals and reduced by half the neurofibrillatory tangles and abnormal accumulations of beta-amyloid. Unfortunately, clioquinol is much too toxic for use in humans.

Beta-amyloid protein in the brain was found to trap copper. The bound copper catalyzed the release of hydrogen peroxide, causing further neurological damage. Accumulations of zinc in the brain acted to create a vicious cycle of increasing beta-amyloid and trapping more copper, leading to progressively more cell damage.

Nutritional and essential trace metallic elements have a very narrow margin between physiologic and toxic levels. Zinc rises to potentially toxic levels in the diseased heart. Chromium and cobalt accumulate even more. It seems likely that other nutritional elements not yet measured may accumulate to a similar extent. Iron accumulates to toxic levels and also acts as a catalyst to greatly speed the production of damaging free oxygen radicals.

We must now carefully consider this recent evidence indicating that EDTA benefits patients, at least in part, by removing abnormal accumulation of essential nutritional trace elements from diseased organs and arterial walls.

EDTA in the body remains extracellular, and can only remove intracellular accumulations of metallic ions by first binding and removing elements outside of cells. That process establishes a strong concentration gradient, which then acts to draw unwanted intracellular metals out through cell walls. Only then can they be chelated. That is one reason why EDTA is administered slowly and intravenously, over several hours. Diffusion outward is a relatively passive process and occurs more slowly than the subsequent binding by EDTA. That may also be the reason why no data have been published showing improved blood flow using oral EDTA. Absorption of oral EDTA is minimal, slow, and plasma concentrations are far lower, resulting in a much weaker intracellular to extracellular concentration gradient.

Based on recent findings, it now seems possible that an important mechanism of action of EDTA chelation therapy is to restore or rebalance safe and desirable levels of essential nutritional metallic elements within cells. A spectrum of such metals has been shown to accumulate to toxic levels in both diseased and stressed cells. Urinary excretion of excessive metals and free radical catalysts might be only a secondary benefit.

BIOFILM ACTION OF EDTA

A strong association between Chlamydia pneumoniae infection and atherosclerosis has been clearly established, although a causal relationship is not agreed upon. 302,303

Infective microorganisms commonly bind tightly together within the body in complex assemblages called biofilm—encased in a matrix of polymeric substances.304 Microorganisms produce this biofilm and become adherent to cell surfaces, which  enhances their survival and reproduction. Mature biofilms consist of large collections of microorganisms embedded in a matrix of non-cellular matter, mineral crystals, and blood components—with channels for diffusion of water, oxygen, and nutrients for bacteria. Microorganisms residing within such biofilms are highly resistant to antibiotics and to normal host immune responses. Up to 60% of human infections and 80% of refractory infections are from bacteria encased in biofilm. Protection thus conferred can allow infections to achieve a high level of antibiotic and immune resistance.

Divalent calcium and magnesium cations bind biofilms together creating an impermeable semi-crystalline matrix. Disodium EDTA binds to calcium and magnesium and thus weakens biofilm matrices. Disodium EDTA also causes structural damage to bacterial cell membranes, making them more permeable to antibiotics. In those ways, chelation therapy can inhibit growth of infective organisms, including those that generate and reside within biofilms. In one in vitro study, disodium EDTA alone reduced the number of biofilm-associated bacteria by up to 99% in a dose-dependent fashion.

This anti-biofilm and anti-infective activity of EDTA is another potential mechanism of action for disodium EDTA chelation therapy.

REFERENCES

1—Clarke NE, Clarke CN, Mosher RE. The “in vivo” dissolution of metastatic calcium: an approach to atherosclerosis. Am J Med Sci 1955;229:142-149.

2—Schroeder HA, Perry HM, Jr. Antihypertensive effects of metal binding agents. J Lab Clin Med 1955;46:416.

3—Clarke NE, Clarke CN, Mosher RE. Treatment of angina pectoris with disodium ethylene diamine tetraacetic acid. Am J Med Sci 1956;232:654-666.

4—Boyle AJ, Casper JJ, McCormick H, et al. Studies in human and induced atherosclerosis employing EDTA. (Swiss, Basel) Bull Schweiz Akad Med Wiss 1957;13:408.

5—Muller SA, Brunsting LA, Winkelmann RK. Treatment of scleroderma with a new chelating agent, edathamil. Arch Dermatol 1959;80:101.

6—Clarke NE, Sr. Atherosclerosis, occlusive vascular disease and EDTA. Am J Cardiol 1960;6:233-236.

7—Meltzer LE, Ural ME, Kitchell JR. The treatment of coronary artery disease with disodium EDTA. In: Seven MJ, Johnson LA, eds. Metal Binding in Medicine Philadelphia: J. B. Lippincott Co; 1960:132-136.

8—Peters HA. Chelation therapy in acute, chronic and mixed porphyria. In: Seven MJ, Johnson LA, eds. Metal Binding in Medicine. Philadelphia: J. B. Lippincott Co; 1960:190-199.

9. Seven MJ, Johnson LA, eds. Metal Binding in Medicine. Proceedings of a Symposium Sponsored by Hahnemann Medical College and Hospital, Philadelphia. Philadelphia: J. B. Lippincott Co; 1960.

10—Kitchell JR, Meltzer LE, Seven MJ. Potential uses of chelation methods in the treatment of cardiovascular diseases. Prog Cardiovasc Dis. 1961;3:338-349.

11—Peters HA. Trace minerals, chelating agents and the porphyrias. Fed Proc 1961;3(2nd pt)(suppl 10):227-234.

12—Boyle AJ, Clarke NE, Mosher RE, et al. Chelation therapy in circulatory and sclerosing diseases. Fed Proc 1961;20(3)(2nd pt) (suppl 10):243-257.

13—Soffer A, Toribara T, Sayman A. Myocardial responses to chelation. Br Heart J 1961 Nov;23:690.

14—Peripheral Flow Opened Up. Medical World News Mar 15, 1963;4:36-39.

15—Boyle AJ, Mosher RE, McCann DS. Some in vivo effects of chelation-I: rheumatoid arthritis. J Chronic Dis 1963;16:325-328.

16—Aronov DM. First experience with the treatment of atherosclerosis patients with calcinosis of the arteries with trilon-B (disodium salt of EDTA). Klin Med (Russ Moscow) 1963;41:19-23.

17—Soffer A, Chenoweth M, Eichhorn G, et al. Chelation Therapy Springfield: Charles C. Thomas; 1964.

18—Soffer A. Chelation therapy for cardiovascular disease. In: Soffer A, ed. Chelation Therapy Springfield: Charles C. Thomas; 1964:15-33.

19—Lamar CP. Chelation therapy for occlusive arteriosclerosis in diabetic patients. Angiology 1964;15:379-394.

20—Friedel W, Schulz FH, Schoder L. Therapy of atherosclerosis through mucopolysaccarides and EDTA (ethylene diamine tetraacetic acid). (German) Deutsch Gesundh 1965;20:1566-1570.

21—Lamar CP. Chelation endarterectomy for occlusive atherosclerosis. J Am Geriatr Soc 1966;14:272-293.

22—Birk RE, Rupe CE. The treatment of systemic sclerosis with EDTA, pyridoxine and reserpine. Henry Ford Hospital Medical Bulletin 1966 June;14:109-139.

23—Lamar CP. Calcium chelation of atherosclerosis, nine years’ clinical experience. Read before the Fourteenth Annual Meeting of the American College of Angiology San Juan, PR, Dec 8, 1968.

24—Olwin JH, Koppel JL. Reduction of elevated plasma lipid levels in atherosclerosis following EDTA chelation therapy. Proc Soc Exp Biol Med 1968;128:1137-1139.

25—Leipzig LJ, Boyle AJ, McCann DS. Case histories of rheumatoid arthritis treated with sodium or magnesium EDTA. J Chronic Dis 1970;22:553-563.

26—Brucknerova O, Tulacek J. Chelates in the treatment of occlusive atherosclerosis. (Czechoslavakian, Praha) Vnitr Lek 1972;18:729-735.

27—Nikitina EK, Abramova MA. Treatment of atherosclerosis patients with Trilon-B (EDTA). (Russian, Moscow) Kardiologiia 1972;12:137-139.

28—Evers R. Chelation of vascular atheromatous disease. Journal International Academy Metabology 1972;2:51-53.

29—Kurliandchikov VN. Treatment of patients with coronary arteriosclerosis with unithiol in combination with vitamins. (Russian, Kiev) Vrach Delo 1973;6:8.

30—Zapadnick VI, et al. Pharmacological activity of unithiol and its use in clinical practice. (Russian, Kiev) Vrach Delo 1973;8:122.

31—David O, Hoffman SP, Sverd J, et al. Lead and hyperactivity, behavioral response to chelation: a pilot study. Am J Psychiatry 1976;133:1155-1158.

32—Gordon GB, Vance RB. EDTA chelation therapy for atherosclerosis: history and mechanisms of action. Osteopathic Annals 1976;4:38-62.

33—Proceedings: Hearing on EDTA Chelation Therapy of the Ad Hoc Scientific Advisory Panel on Internal Medicine of the Scientific Board of the California Medical Society, March 26, 1976, San Francisco, California.

34—Halstead BW. The Scientific Basis of EDTA Chelation Therapy Colton, CA: Golden Quill Publishers; 1979.

35—Grumbles LA. Radionuclide Studies of Cerebral and Cardiac Circulation Before and After Chelation Therapy. Presented at a meeting of the American Academy of Medical Preventics, Chicago, Illinois, May 27, 1979.

36—Bjorksten J. The cross-linkage theory of aging as a predictive indicator. Journal of Advancement in Medicine 1989;2(1&2):59-76.

37—Bjorksten J. Possibilities and limitations of chelation as a means for life extension. Journal of Advancement in Medicine 1989;2(1&2):77-88.

38—Blumer W, Cranton EM. Ninety percent reduction in cancer mortality after chelation therapy with EDTA. Journal of Advancement in Medicine 1989;2(1&2):183-188.

39—Carpenter DG. Correction of biological aging. Rejuvenation 1980;8:31-49

40—Casdorph HR. EDTA chelation therapy, efficacy in arteriosclerotic heart disease. Journal of Advancement in Medicine 1989;2(1&2):121-130.

41—Casdorph HR. EDTA chelation therapy II, efficacy in brain disorders. Journal of Advancement in Medicine 1989;2(1&2):131-154.

42—McDonagh EW, Rudolph CJ, Cheraskin E. An oculocerebrovasculometric analysis of the improvement in arterial stenosis following EDTA chelation therapy. Journal of Advancement in Medicine 1989;2(1&2):121-130.

43—Olwin JH. EDTA Chelation Therapy. Presented at a meeting of the American Holistic Medical Association, University of Wisconsin, La Crosse, Wisconsin, May 28, 1981.

44—McDonagh EW, Rudolph CJ, Cheraskin E. The influence of EDTA salts plus multi-vitamin-trace mineral therapy upon total serum cholesterol/high-density lipoprotein cholesterol. Medical Hypothesis 1982;9:643-646.

45—McDonagh EW, Rudolph CJ, Cheraskin E. The effect of intravenous disodium ethylenediaminetetraacetic acid (EDTA) upon blood cholesterol in a private practice environment. Journal of the International Academy of Preventive Medicine 1982;7:5-12.

46—Williams DR, Halstead BW. Chelating agents in medicine. J Toxicol: Clin Toxicol 1983;19(10):1081-1115.

47—Casodorph HR, Farr CH. EDTA chelation therapy: treatment of peripheral arterial occlusion, an alternative to amputation. Journal of Advancement in Medicine 1989;2(1&2):167-182.

48—McDonagh EW, Rudolph CJ, Cheraskin E. The effect of EDTA chelation therapy with multivitamin/trace mineral supplementation upon reported fatigue. Journal of Orthomolecular Psychiatry 1984;13(4):277-279.

49—Riordan HD, Jackson JA, Cheraskin E. The effects of intravenous EDTA infusion on the multichemical profile. American Clinical Laboratory Oct 1988.

50—Riordan HD, Cheraskin E, Dirks M, et al: Electrocardiographic changes associated with EDTA chelation therapy. Journal of Advancement in Medicine 1988;1(4):191-194.

51—Deucher GP. EDTA chelation therapy: an antioxidant strategy. Journal of Advancement in Medicine 1988;1(4):182-190.

52—Rudolph CJ, McDonagh EW, Wussow DG. The effect of intravenous disodium ethylenediaminetetaacetic acid (EDTA) upon bone density. Journal of Advancement in Medicine 1988;1(2):79-85.

53—McDonagh EW, Rudolph DO, Cheraskin E. Effect of chelation therapy plus multivitamin/mineral supplementation upon vascular dynamics: ankle/brachial doppler blood pressure ratio. Journal of Advancement in Medicine 1989;2(1&2):159-166.

54—McDonagh EW, Rudolph DO, Cheraskin E. The “clinical change” in patients treated with EDTA chelation plus multivitamin/mineral supplemention. Journal of Advancement in Medicine 1989;2(1&2):189-196.

55—Olzewer E, Carter JP. EDTA chelation therapy: a retrospective study of 2,870 patients. Journal of Advancement in Medicine 1989;2(1&2):197-213.

56—Rudolph CJ, McDonagh EW, Barber RK. A non-surgical approach to obstructive carotid stenosis using EDTA chelation. Journal of Advancement in Medicine 1991;4(3):157-165.

57—Chappell LT, Stahl JP. The correlation between EDTA chelation therapy and improvement in cardiovascular function: a meta-analysis. Journal of Advancement in Medicine 1993;6(3):139-160.

58—Chappell LT, Stahl JP, Evans R. EDTA chelation treatment for vascular disease: a meta-analysis using unpublished data. Journal of Advancement in Medicine 1994;7(3):131-142.

59— Hancke C, Flytlie K. Benefits of EDTA chelation therapy on atherosclerosis: a retrospective study of 470 patients. Journal of Advancement in Medicine 1993;6(3):61-171.

60—Rudolph CJ, Samuels RT, McDonagh EW. Visual field evidence of macular degeneration reversal using a combination of EDTA chelation and multiple vitamin and trace mineral therapy. Journal of Advancement in Medicine 1994;7(4):203-212.

61—Holliday HJ. Carotid restenosis: a case for EDTA chelation. Journal of Advancement in Medicine 1996;9(2):95-100.

62—Olwin JH, Kannabrocki EL, Sothern RB. Rationale for the use of EDTA-magnesium-heparin therapy in subjects with coronary artery disease. Journal of Advancement in Medicine 1997;10(2):157-165.

63—Olszewer E, Sabbag FC, Carter JP. A pilot double-blind study of sodium-magnesium EDTA in peripheral vascular disease. J Natl Med Assn 1990;82(3):174-177.

64—Doolan PD, Schwartz SL, Hayes JR, et al. An evaluation of the nephro¬ toxicity of ethylenediaminetetraacetate and diethylenetriaminepentaacetate in the rat. Toxicol Appl Pharmacol 1967;10:481-500.

65—Ahrens FA, Aronson AL: A comparative study of the toxic effects of calcium and chromium chelates of ethylenediaminetetraacetate in the dog. Toxicol Appl Pharmacol 1971;18:10-25.

66—Feldman EB: EDTA and angina pectoris. Drug Therapy 1975 Mar:62.

67—Wedeen RP, Mallik DK, Batuman V. Detection and treatment of occupational lead nephropathy. Arch Intern Med 1979,139:53-57.

68—Moel DI, Kuman K. Reversible nephrotoxic reactions to a combined 2, 3-dimercapto-1-propanol and calcium disodium ethylenediaminetetraacetic acid regimen in asymptomatic children with elevated blood levels. Pediatrics 1982;70(2):259-262.

69—McDonagh EW, Rudolph CJ, Cheraskin E. The effect of EDTA chelation therapy plus supportive multivitamin-trace mineral supplementation upon renal function: a study in serum creatinine. Journal of Advancement in Medicine 1989,2(1&2):235-243.

70—Cranton EM. Kidney effects of ethylene diamine tetraacetic acid (EDTA): a literature review. Journal of Advancement in Medicine 1989;2(1&2):227-233.

71—Batuman V, Landy E, Maesaka JK, et al. Contribution of lead to hyper¬ tension with renal impairment. N Engl J Med 1983;309(1):17-21.

72—McDonagh EW, Rudolph CJ, Cheraskin E. The effect of EDTA chelation therapy plus supportive multivitamin-trace mineral supplementation upon renal function: a study in blood urea nitrogen (BUN), J Holistic Med 1989;2(1&2):251-261.

73—Sehnert KW, Claque AF, and Cheraskin E. The improvement in renal function following EDTA chelation and multivitamin trace mineral therapy: a study in creatinine clearance. Journal of Advancement in Medicine 1989;2(1&2):245-250.

74—Riordon HD, Cheraskin E, Dirks M, et al. Another look at renal function and the EDTA treatment process. Journal of Advancement in Medicine 1989;2(1&2):263-268.

75—Kitchell JR, Palmon F, Aytan N, et al. The treatment of coronary artery disease with disodium EDTA, a reappraisal. Am J Cardiol 1963;11:501-506.

76—Cranton EM, Frackelton JP. Current status of EDTA chelation therapy in occlusive arterial disease. Journal of Advancement in Medicine 1989;2(1&2):107-119.

77—Wartman A, Lampe TL, McCann DS, et al. Plaque reversal with MgEDTA in experimental atherosclerosis: elastin and collagen metabolism. J Atheros Res 1967;7:331.

78—Wissler RW: Principles of the pathogenesis of atherosclerosis. In: Braunwald E, ed. Heart Disease Philadelphia: W. B. Saunders Co; 1980:1221-1236.

79—Kjeldsen K, Astrup P, Wanstrup J. Reversal of rabbit atherosclerosis by hyperoxia. J Atheros Res 1969;10:173.

80—Vesselinovitch D, Wissler RW, Fischer-Dzoga K, et al. Regression of atherosclerosis in rabbits. I. Treatment with low fat diet, hyperoxia and hypolipidemic agents. Atherosclerosis 1974;19:259.

81—Sincock A. Life extension in the rotifer by application of chelating agents. J Gerontot 1975;30:289-293.

82—Wissler RW, Vesselinovitch D. Regression of atherosclerosis in experimental animals and man. Mod Concepts Cardiovasc Dis 1977,46:28.

83—Walker F. The Effects of EDTA Chelation Therapy on Plaque, Calcium, and Mineral Metabolism in Arteriosclerotic Rabbits. Ph.D. thesis. Texas State University, 1980. (Available from University Microfilm International, Ann Arbor, MI 48016.)

84—Guldager B, Jelnes R, Jorgensen SJ, et al. EDTA treatment of intermittent claudication—a double-blind, placebo-controlled study. Journal of Internal Medicine 1992;231:261-267.

85—Sloth-Nielsen J, Guldager B. Mouritzen C. Arteriographic findings in EDTA chelation therapy on peripheral atherosclerosis. The American Journal of Surgery. 1991;162:122-125.

86—Van Rij AM, Solomon C, Packer SG. Chelation therapy for intermittent claudication. A double-blind, randomized, controlled study. Circulation. 1994 Sep;90(3):1194-1199.

87—Diehm C, Wilhelm C, Poeschl J. Effects of EDTA-Chelation Therapy in Patients with Peripheral Vascular Disease—A Double-Blind Study. An unpublished study performed by the Department of Internal Medicine, University of Heidelberg, Heidelberg, Germany in 1985. Presented as a paper before the International Symposium of Atherosclerosis, Melbourne, Australia, October 14, 1985.

88—Carter JP. If EDTA chelation therapy is so good, why is it not more widely accepted? Journal of Advancement in Medicine 1989;2(1&2):213-226.

89—Chappell LT. Disputes author’s conclusions on effectiveness of EDTA chelation therapy. Alternative Therapies. Sep 1996;2(5):16-17.

89A—Lamas GA, Goertz C, Boineau R, et al: Design of the Trial to Assess Chelation Therapy (TACT). Am Heart J. 2012 Jan;163(1):7-12.

89B—Lamas GA, Goertz C, Boineau R, TACT Investigators, et al: Effect of disodium EDTA chelation regimen on cardiovascular events in patients with previous myocardial infarction: the TACT randomized trial. JAMA. 2013 Mar 27;309(12):1241-50.

90—Fossel M. Role of cell senescence in human aging. Journal of Anti-Aging Medicine 2000;3(1):91-98.

91—Fossel M. Telomerase and the aging cell: implications for human health. JAMA 1998;279:1732-1735.

92—Fossel M. Reversing Human Aging. New York: William Morrow and Company; 1996.

93—Benchimol S, Vaziri H. Reconstruction of telomerase activity in normal cells leads to elongation of telomeres and extended replicative life span. Curr Biol 1998;8:279-282.

94—Chang E, Harley CB. Telomere length as a measure of replicative histories in human vascular tissues. Proc Natl Acad Sci USA 1995;92:1190-1194

95—Frustaci A, Magnavita N, Chirmenti C, et al. Marked elevation of myocardial trace elements in idiopathic dilated cardiomyopathy compared with secondary cardiac dysfunction. JACC 1999;33(6):1578-1583.

96—Tatton WG, Olanow CW. Apoptosis in neurodegenerative diseases: the role of mitochondria. Biochem Biophy Acta 1999;1410(2):195-213 (a review).

96A —Ong SB, Hall AR, Hausenloy DJ. Mitochondrial Dynamics in Cardiovascular Health and Disease. Antioxid Redox Signal. 2012 Jul 15. [PubMed. Epub ahead of print]

97—Peng L, Nijjhawan D, Budihardya SM, et al. Cytochrome c- and d-ATP-dependent Apaf-l/caspase-9 complex initiate an apoptotic protease cascade. Cell 1997;91:479-489.

98—Demopoulos HB, Pietronigro DD, Flamm ES, et al. The possible role of free radical reactions in carcinogenesis. Journal of Environmental Pathology and Toxicology 1980;3:273-303.

98A —Jomova K, Valko M. Advances in metal-induced oxidative stress and human disease. Toxicology. 2011 May 10;283(2-3):65-87. Epub 2011 Mar 23.

99—Harman D. The aging process. Proc Natl Acad Sci USA 1981;78:7124-7128.

100—Dormandy TL. An approach to free radicals. Lancet 1983,ii.1010-14.

101—Demopoulos HB, Pietronigro DD, Seligman ML. The development of secondary pathology with free radical reactions as a threshold mechanism. Journal of the American College of Toxicology 1983;2(3):173-184.

102—Demopoulos HB. Molecular Oxygen in Health and Disease. Read before the American Academy of Medical Preventics Tenth Annual Spring Conference, Los Angeles, California May 21, 1983.(Available on three audio cassettes from Instatape, P.O. Box 1729, Monrovia, CA 91016.)

103—Ames BN. Dietary carcinogens and anticarcinogens. Science. 1983;221:1256-1264.

104—Dormandy TL. Free-radical reaction in biological systems. Ann R Coll Surg Engl 1980;62:188-194.

105—Dormandy TL. Free-radical oxidation and antioxidants. Lancet 1978;8:647-650.

106—Levine SA, Reinhardt JH. Biochemical-pathology initiated by free radicals, oxidant chemicals, and therapeutic drugs in the etiology of chemical hypersensitivity disease. Journal of Orthomolecular Psychiatry 1983;12(3):166-183.

106A—Martin-Ventura JL, Madrigal-Matute J, Martinez-Pinna R, Ramos-Mozo P, Blanco-Colio LM, Moreno JA, Tarin C, Burillo E, Fernandez-Garcia CE, Egido J, Meilhac O, Michel JB. Erythrocytes, leukocytes and platelets as a source of oxidative stress in chronic vascular diseases: Detoxifying mechanisms and potential therapeutic options. Thromb Haemost. 2012 Jul 26;108(3). [Epub ahead of print]

107—Del Maestro RF. An approach to free radicals in medicine and biology. Acta Physiol Scand 1980;492(supp1):153-68.

108—Poole CP. Electron Spin Resonance A Comprehensive Treatise on Experimental Techniques. New York: Interscience Publishers; 1967.

109—Demopoulos HB, Flamm ES, Seligman ML, et al. Membrane perturbations in central nervous system injury: theoretical basis for free radical damage and a review of the experimental data. In: Popp AJ, Bourke LR, Nelson LR, Kimelbert HK, eds. Neural Trauma. New York: Raven Press; 1979:63-78.

110—Seligman ML, Mitamura JA, Shera N, et al. Corticosteroid (methylprednisolone) modulation of photoperoxidation by ultra¬ violet light in liposomes. Photochem Photobiol 1979;29:549-558.

111—Pryor WA. Free radical reactions and their importance in biochemical systems. Fed Proc 1973;32:1862-1869.

112—Pryor WA, ed. Free Radicals in Biology Volumes 1-3. New York: Academic Press; 1976.

113—Lambert L, Willis ED. The effect of dietary lipid peroxides, sterols and oxidized sterols on cytochrome P450 and oxidative demethylation. Biochem Pharmacol 1977a;26:1417-1421.

113A—Wonisch W, Falk A, Sundl I, Winklhofer-Roob BM, Lindschinger M. Oxidative stress increases continuously with BMI and age with unfavourable profiles in males. Aging Male. 2012 Apr 2. [Epub ahead of print]

114—Lambert C, Willis ED. The effect of dietary lipids on 3, 4 benzo(a)pyrene metabolism in the hepatic endoplasmic reticulum. Biochem Pharmacol 1977b;26:1423-1477.

115—Fedorenko VI. Effect of cysteine, glutathione and 1-p-chlorophenyltetrazole-thione-2 on postradiation changes in the metabolic free radical content of albino rat tissues. Radiobiologiia 1979;19:67-73.

116—Fridovich I. Superoxide dismutases. Annu Rev Biochem 1975:147-159.

117—Black HS, Chan JT. Experimental ultraviolet light-carcinogenesis. Photochem Photobiol 1977;26:183-189.

118—Eaton GJ, Custer P, Crane R. Effects of ultraviolet light on nude mice: cutaneous carcinogenesis and possible leukemogenesis. Cancer 1978;42:182-188.

119—Tappel AL. Lipid peroxidation damage to cell components. Fed Proc 1973;32:1870-1874.

120—Kotin P, Falk HL. Organic peroxide, hydrogen peroxide, epoxides and neoplasia. Radiat Res 1963;3(supp1):193-211.

121. —Demopoulos, HB. Control of free radicals in the biologic systems. Fed Proc. 1973;32:1903-1908.

122—Walling C. Forty years of free radicals. In: Pryor WA, ed. Organic Free Radicals Washington, DC: American Chemical Society; 1978.

123—Demopoulos HB. The basis of free radical pathology. Fed Proc 1973;32:1859-1861.

124—Tappel AL. Will antioxidant nutrients slow aging processes? Geriatrics 1968;23:97-105.

124A—Box HC, Patrzyc HB, Budzinski EE, Dawidzik JB, Freund HG, Zeitouni NC, Mahoney MC. Profiling Oxidative DNA Damage: Effects of Antioxidants. Cancer Sci. 2012 Jul 26. doi: 10.1111/j.1349-7006.2012.02391.x. [Epub ahead of print]

125—Coon MJ. Oxygen activation in the metabolism of lipids, drugs and carcinogens. Nutr Rev 1978;36:319-328.

126—Coon MJ. Reconstitution of the cytochrome P-450-containing mixed-function oxidase system of liver microsomes. Methods Enzymol 1978;52:200-206.

127—Coon MJ, van der Hoeven TA, Dahl SB, et al. Two forms of liver microsomal cytochrome P-450, P-4501m2 and P-450M4 (rabbit liver). Methods Enzymol 1978;52:109-117.

128—Panganamala RV, Sharma HM, Sprecher H, et al. A suggested role for hydrogen peroxide in the biosynthesis of prostaglandins. Prostaglandins 1974;8:3-11.

129—Maisin JR, Decleve A, Gerber GB, et al. Chemical protection against the long-term effects of a single whole-body exposure of mice to ionizing radiation. II. Causes of death. Radiat Res 1978;74:415-435.

130—McGinnes JE, Proctor PH, Demopoulos HB, et al. In vivo evidence for superoxide and peroxide production by adriamycin and cis-platium. In: Autor A, ed. Active Oxygen and Medicine New York: Raven Press; 1980.

131—Petkau A. Radiation protection by superoxide dismutase. Photochem Photobiol 1978;28:765-774.

132—Schaefer A, Komlos M, Seregi A. Lipid peroxidation as the cause of the ascorbic acid induced decrease of ATPase activities of rat brain microsomes and its inhibition by biogenic amines and psychotropic drugs. Biochem Pharmcol 1975;24: 1781-1786.

133—Vladimirov YA, Sergeer PV, Seifulla RD, et al. Effect of steroids on lipid peroxidation and liver mitochondrial membranes. Molekuliarnaia Biologiia (Russian, Moscow) 1973;7:247-262. (Translated by Consultants Bureau, a division of Plenum Publishing Inc., New York.)

134—Sies H, Summer KH. Hydroperoxide-metabilizing systems in rat liver. Eur J Biochem 1975;57:503-512.

135—Alfthan G, Pikkarainen J, Huttunen JK, et al. Association between cardiovascular death and myocardial infarction and serum selenium in a matched-pair longitudinal study. Lancet 1982;2(8291):175-179.

135A—Tanguy S, Grauzam S, de Leiris J, Boucher F. Impact of dietary selenium intake on cardiac health: Experimental approaches and human studies. Mol Nutr Food Res. 2012 Jul;56(7):1106-21. doi: 10.1002/mnfr.201100766.

136—Willett WC, Morris JS, Pressel S, et al. Prediagnostic serum selenium and risk of cancer. Lancet 1983;2(8343):130-134.

137—Demopoulos HB, Flamm ES, Seligman ML, et al. Antioxidant effects of barbiturates in model membranes undergoing free radical damage. Acta Neurol Scand 1977;56(suppl 64):152.

138—Flamm ES, Demopoulos HB, Seligman ML, et al. Barbiturates and free radicals. In: Popp AJ, Popp RS, Bourke LR, Nelson HB, Kimelberg HK, eds. Neural Trauma. New York: Raven Press; 1979:289-300.

139—Butterfield JD, McGraw CP. Free radical pathology. Stroke 1978;9(5):443-445.

140—Demopoulos HB, Flamm ES, Pietronigro DD, et al. The free radical pathology and the microcirculation in the major ¬ central nervous system disorders. Acta Physiol Scand 1980;492(suppl):91-119.

141—Gey KF, Puska P, Jordan P, et al. Inverse correlation between plasma vitamin E and mortality from ischemic heart disease in cross-cultural epidemiology. Am J Clin Nutr 1991;53:326S-334S.

142—Seubens WE, Smith S. Serum cholesterol correlations with atherosclerosis at autopsy. American Clinical Laboratory 1997 Apr;14-15.

143—Morel DW, Hessler JR, Chisolm GM. Low-density lipoprotein cytotoxicity induced by free radical peroxidation of lipid. Journal of Lipid Research 1983;24:1070-1076.

144—Smith LL. Cholesterol Autooxidation. New York: Plenum Press; 1981.

145—Gaby AR. Nutritional factors in cardiovascular disease. Journal of Advancement in Medicine 1989;2(1&2):89-105.

146—Taylor CB, Peng SK, Werthessen NT, et al. Spontaneously occurring angiotoxic derivatives of cholesterol. Am J Clin Nutr 1979;32:40.

147—Cortese C, Bernardini S, Motti C. Atherosclerosis in light of the evidence from large statin trials. Ann Ital Med Int 2000;15(1):103-107

148—Song YM, Sung J, Kim JS. Which cholesterol level is related to the lowest mortality in a population with low mean cholesterol level: a 6.4-year follow-up study of 482,472 Korean men. Am J Epidemiol 2000;151(8):739-747.

149—Wilcken DEL, Wilcken B. The pathogenesis of coronary artery disease: a possible role for methionine metabolism. J Clin Investig 1976;57:1079-1082.

150—Boushey CJ, Beresford SAA, Omenn GS, et al. A quantitative assessment of plasma homocysteine as a risk factor for vascular disease: probable benefits of increasing folic acid intakes. JAMA 1995;274:1049-1057.

151—Clarke R, Daly LE, Robinson K, et al. Hyperhomocysteinernia: an independent risk factor for vascular disease. N Engl J Med 1991;324:1149-1155.

152—Graham IM, Daly LE, Refsum HM, et al. Plasma homocysteine as a risk factor for vascular disease: the European concerted action project. JAMA 1997;277:1775-1781.

153—Nygard O, Vollsett SE, Refsum HM, et al. Total plasma homocysteine and cardiovascular risk profile: the Hordaland hornocysteine study. JAMA 1995;274:1526-1533.

154—Clopath P, Smith VC, McCully KS. Growth promotion by homocysteic acid. Science 1976;192:372-374.

155—Tsai J-C, Perella MA, Yoshizumi M, et al. Promotion of vascular smooth muscle cell growth by homocysteine: a link to atherosclerosis. Proc Natl Acad Sci USA 1994;91:6369-6373.

156—Welch GN, Upchurch GR, Loscalzo J. Homocysteine, oxidative stress, and vascular disease. Hosp Pract 1997;32:81-92.

157—D’Angelo A, Selhub J. Homocysteine and thrombotie disease. Blood 1997;90:1-11.

158—Naruszewicz M, Mirkiewicz E, Olszewski AJ, et al. Thiolation of low-density lipoprotein by homocysteine thiolactone causes increased aggregation and altered interaction with cultured macrophages. Nutr Metab Cardiovasc Dis 1994;4:70-77.

159—McCully KS. Homocysteine and vascular disease. Nat Med 1996;2:386-389.

159A—Marosi K, Agota A, Végh V, Joó JG, Langmár Z, Kriszbacher I, Nagy ZB. The role of homocysteine and methylenetetrahydrofolate reductase, methionine synthase, methionine synthase reductase polymorphisms in the development of cardiovascular diseases and hypertension. 2012 Mar 25;153(12):445-53.

160—McCully KS. Homocysteine, folate, vitamin B-6, and cardiovascular disease. Editorial. JAMA 1998;279(5):392-393.

161—McCully KS. Vascular pathology of hornocysteinernia: implications for the pathogenesis of arteriosclerosis. Am J Pathol 1969;56:111-128.

162—Rimm EB, Willett WC, Hu FB, et al. Folate and vitamin B-6 from diet and supplements in relation to risk of coronary heart disease among women. JAMA 1998;279(5):359-364.

163—Boushey CJ, Beresford SAA, Omenn GS, et al. A quantitative assessment of plasma homocysteine as a risk factor for vascular disease: probable benefits of increasing folic acid intakes. JAMA 1996;274:1049-1057.

164—Brattstrom L, Israelsson B, Norrving B, et al. Impaired homocysteine metabolism in early onset cerebral and peripheral occlusive arterial disease: effects of pyridoxine and folic acid treatment. Atherosclerosis 1990;81:51-60.

165—Jacob RA, Wu M, Henning SM, et al. Homocysteine increases as folate decreases in plasma of healthy men during short-term dietary folate and methyl group restriction. J Nutr 1994;124:1072-1080.

166—Naurath HJ, Joosten E, Riezler R, et al. Effects of vitamin B-12, folate, and vitamin B-6 supplements in elderly people with normal serum vitamin concentrations. Lancet 1996,346:85-89.

167—Selhub J, Jacques PF, Bostom AG, et al. Association between plasma homocysteine concentrations and extracranial carotid-artery stenosis. N Engl J Med 1995;332:289-291.

168—Nygard O, Nordrehaug JE, Refsurn H, et al. Plasma homocysteine levels and mortality in patients with coronary artery disease. N Engl J Med 1997;337:230-236.

169—Rimin EB, Stampfer MJ, Ascherio A, et al. Dietary folate, vitamin B-6, and vitamin B-12 intake and risk of CHD among a large population of men. Abstract. Circulation 1996;93:625.

170—Passwater RA, Cranton EM. Trace Elements, Hair Analysis and Nutrition. New Canaan, Ct: Keats Publishing, Inc; 1983.

171—Fourcans B. Role of phospholipids in transport and enzymic reactions. Adu Lipid Res 1974:147-226.

172—Babior BM. Oxygen-dependent microbial killing by phagocytes. N Engl J Med 1978;298:659-668.

173—Rosen H. Klebanoff SJ. Bactericidal activity of a superoxide anion-generating system. J Exp Med 1979;149:27-39.

174—Masterson WL, Slowinski E. In: Chemical Principles. Philadelphia: Saunders; 1977:203(plate 5).

175—Mayes PA. Biologic oxidation. In: Martin DW, Mayes PA, Rodwell VW, eds. Harper’s Review of Biochemistry. Los Alton, Ca: Lange Medical Publications; 1983:129-130.

176—March J. Advanced Organic Chemistry Reactions, Mechanics, and Structure. 2nd ed. New York: McGraw-Hill; 1978:620.

177—Flamm ES, Demopoulos HB, Seligman ML, et al. Free radicals in cerebral ischemia. Stroke 1978;9(5):445-447.

177A—Wang CX, Shuaib A.Neuroprotective effects of free radical scavengers in stroke. Drugs Aging. 2007;24(7):537-46.

178—Hyperbaric Oxygen Therapy: A Committee Report. February 1981. Undersea Medical Society, Inc., 9650 Rockville Pike, Bethesda, MD 20014 (UMS Publication Number 30 CR(HBO) 2-23-81).

179—Foote CS. Chemistry of singlet oxygen VII. Quenching by beta carotene. J Am Chem Soc 1968;90:6233.

180—Rimm EB, Stampfer MJ, Ascheno A, et al. Vitamin E consumption and the risk of coronary heart disease in men. N Engl J Med 1993 May 20;328(20):1450-1456.

181—Stampher MJ, Hennekens CH, Morrison JE, et al. Vitamin E consumption and the risk of coronary disease in women. N Engl J Med 1993 May 20;328(20):1444-1449.

182—Rimm EB, Stampfer MJ, Ascheno A, et al. Vitamin E consumption and the risk of coronary heart disease in men. N Engl J Med 1993 May 20;328(20):1450-1456.

183—Peto R, Doll R, Buckley JD, et al. Can dietary beta-carotene materially reduce human cancer rates? Nature 1981;290:201.

184—Stoyanovsky DA, et al. Endogenous ascorbate regenerates vitamin E in the retina directly and in combination with exogenous dihydrolipoic acid. Curr Eye Res 1995 Mar;14(3):181-189.

185—Chan AC. Partners in defense, vitamin E and vitamin C. Can J Physiol Pharmacol 1993 Sep;71(9):725-731.

186—Ho CT, et al. Regeneration of vitamin E in rat polymorphonuclear leucocytes. FEBS Lett 1992 Jul 20;306(2-3):269-272.

187—Chan AC, et al. Regeneration of vitamin E in human platelets. J Biol Chem 1991 Sep 15;266(26):17290-17295.

188—Tappel AL. Will antioxidant nutrients slow aging processes? Geriatrics 1968 Oct;23(10):97-105.

189—Enstrom EE, Kanim LE, Klein MA. Vitamin C intake and mortality among a sample of the United States population. Epidemiology 1992;3(3):194-202.

190—Schaefer A, Komlos M, Seregi A. Lipid peroxidation as the cause of the ascorbic acid induced decrease of ATPase activities of rat brain microsomes and its inhibition by biogenic amines and psychotropic drugs. Biochem Pharmacol 1975;24:1781-86.

191—Ito T, Allen N, Yashon D. A mitochondrial lesion in experimental spinal cord trauma. J Neurosurg 1978;48:434-442.

192—Gillman MW, Cupples LA, Gagnon D, et al. Margarine intake and subsequent coronary heart disease in men. Epidemiology 1997 Mar;8(2):144-149.

193—Atherosclerosis and auto-oxidation of cholesterol. Lancet 1980;ii:964-965.

194—Pietronigro DD, Demopoulos HB, Hovsepian M, et al. Brain ascorbic acid (AA) depletion during cerebral ischemia. Stroke 1982;13(1):117.

195—Demopoulos HB, Flamm ES, Seligman ML, et al. Further studies on free-radical pathology in the major central nervous system disorders: effect of very high doses of methylprednisolone on the functional outcome; morphology and chemistry of experimental spinal cord impact injury. Can J Physiol Pharmacol 1982;60(11):1415-1424.

196—Flamm ES, Demopoulos HB, Seligman ML, et al. Free radicals in cerebral ischemia. Stroke 1978;9:445.

197—Demopoulos HB, Flamm ES, Seligman ML, et al. Oxygen free radicals in central nervous system ischemia and trauma. In: Autor AP, ed. Pathology of Oxygen New York: Academic Press; 1982:127-155.

198—Demopoulos HB, Flamm ES, Seligman ML, et al. Molecular pathogenesis of spinal cord degeneration after traumatic injury. In: Naftchi NE, ed. Spinal Cord Injury New York and London: Spectrum Publications, Inc; 1982:45-64.

199—Sukoff MH, Hollin SA, Espinosa OK, et al. The protective effect of hyperbaric oxygenation in experimental cerebral edema. J Neurosur 1968;29:236-239.

200—Kelly DL Jr, Lassiter KRL, Vongsvivut A, et al. Effects of hyperbaric oxygen and tissue oxygen studies in experimental paraplegia. J Neurosurg 1972;36:425-429.

201—Holbach KH, Wassman H, Hoheluchter KL, et al. Clinical course of spinal lesions treated with hyperbaric oxygen. Acta Neurochir 1975;31:297-298.

202—Holbach KH, Wassman H, Linke D. The use of hyperbaric oxygenation in the treatment of spinal cord lesions. Eur Neurol 1977;16:213-221.

203—Yeo JD, Stabback S, McKinsey B. Study of the effects of hyperbaric oxygenation on experimental spinal cord injury. Med J Aust 1977;2:145-147.

204—Jones RF, Unsworth IP, Marasszeky JE. Hyperbaric oxygen and acute spinal cord injuries in humans. Med J Aust 1978;2:573-575.

205—Yeo JD, Lawry C. Preliminary report on ten patients with spinal cord injuries treated with hyperbaric oxygenation. Med J Aust 1978,2:572-573.

206—Gelderd JB, Welch DW, Fife WP, et al. Therapeutic effects of hyperbaric oxygen and dimethyl sulfoxide following spinal cord transections in rats. Undersea Biomedical Research 1980;7:305-320.

207—Sukoff MH. Central nervous system: review and update cerebral edema and spinal cord injuries. HBO Review 1980,1:189-195.

208—Jesus-Greenberg DA. Acute spinal cord injury and hyperbaric oxygen therapy: a new adjunct in management. Journal of Neurosurgical Nursing 1980:12:155-160.

209—Higgins AC, Pearlstein MS, Mullen JB, et al. Effects of hyperbaric oxygen therapy on long-tract neuronal conduction in the acute phase of spinal cord injury. J Neurosurg 1981;55(4):501-510.

210—Sukoff MH, Ragatz RE. Use of hyperbaric oxygen for acute cerebral edema. Neurosurgery 1982;10:29-38.

211—De La Torre JC, Johnson CM, Goode DJ, et al. Pharmacologic treatment and evaluation of permanent experimental spinal cord trauma. Neurology 1975;25:508-514.

212—De La Torre JC, Kawanaga HM, Rowed DW, et al. Dimethyl sulfoxide in central nervous system trauma. Ann NY Acad Sci 1975,243:362-389.

213—De La Torre JC, Surgeon JW. Dexamethasone and DMSO in experimental transorbital cerebral infarction. Stroke 1976,7:577-583.

214—Laha RK, Dujovny M, Barrionuevo PJ, et al. Protective effects of methyl prednisolone and dimethyl sulfoxide in experimental middle cerebral artery embolectomy. J Neurosurg 1978;49:508-516.

215—Fischer BH, Marks M, Reich T. Hyperbaric-oxygen treatment of multiple sclerosis. N Eng J Med 1983;308:181-186.

216—Swank R. A Biochemical Basis of Multiple Sclerosis. Springfield, Ill: Charles C. Thomas; 1961.

217—Cimino JA, Demopoulos HB. Introduction: determinants of cancer relevant to prevention, in the war on cancer. Journal of Environmental Pathology and Toxicology 1980;3:1-10.

218—Seligman ML, Flamm ES, Goldstein BD, et al. Spectro¬ fluore¬ scent detection of malonaldehyde as a measure of lipid free radical damage in response to ethanol potentiation of spinal cord trauma. Lipids 1977;12(11):945-950.

219—Flamm ES, Demopoulos HB, Seligman ML, et al. Ethanol potentiation of central nervous system trauma. J Neurosurg 1977;46:328-334.

220—Dix T. Metabolism of polycyclic aromatic hydrocarbon derivatives to ultimate carcinogens during lipid peroxidation. Science 1983;221:277.

221—Samuelsson B. Leukotrienes: mediators of immediate hypersensitivity reactions and inflammation. Science 1983;220:568-575

222—Hess ML, Manson NH, Okabe E. Involvement of free radicals in the pathophysiology of ischemic heart disease. Can J Physiol Pharmacol 1982;60(11):1382-1389.

223—Vincent GM, Anderson JL, Marshall HW. Coronary spasm producing coronary thrombosis and myocardial infarction. N Engl J Med 1983;309(14):220-239.

224—Harmon D. The Free Radical Theory of Aging. Read before the Orthomolecular Medical Society, San Francisco, California, May 8, 1983. (Available on audio cassette from AUDIO-STATS, 3221 Carter Avenue, Marina Del Rey, CA 90291.)

225—Singal PK, Kapur N, Dhillon KS, et al. Role of free radicals in catecholamine-induced cardiomyopathy. Can J Physiol Pharmacol 198260(11):1340-1397.

226—Crapper-McLaughlin DR. Aluminum Toxicity in Senile Dementia: Implications for Treatment. Read before the Fall Conference, American Academy of Medical Preventics, Las Vegas, Nevada, Nov 8, 1981.

227—Raymond JP, Merceron R, Isaac R, et al. Effects of EDTA and Hypercalcemia on Plasma Prolactin, Parathyroid Hormone and Calcitonin in Normal and Parathyroidectomized Individuals. Read before the Frances and Anthony D’Anna International Memorial Symposium, Clinical Disorders of Bone and Mineral Metabolism, May 8, 1983. (Abstract available from Henry Ford Hospital, Dearborn, Michigan.)

228—Frost HB. Coherence treatment of osteoporosis. Orthop Clin North Am 1981;12:649-669.

229—DeLuca HF, Frost HM, Jee WSS, et al, eds. Osteoporosis: Recent Advances in Pathogenesis and Treatment. Baltimore: University Press; 1981.

230—Frost HM. Treatment of osteoporosis by manipulation of coherent bone cell populations. Clin Orthop 1979;143:227-244.

231—Meyer MS, Chalmers TM, Reynolds JJ. Inhibitory Effect of Follicular Stimulating Hormone in Parathormone in Rat Calvaria In Vitro. Read before the Frances and Anthony D’Anna International Memorial Symposium, Clinical Disorders of Bone and Mineral Metabolism, May 8, 1983. (Abstract available from Henry Ford Hospital, Dearborn, Michigan.)

232—Wills ED. Lipid peroxide formation in microsomes. Biochem J 1969;113:325-332.

233—Gutteridge JMC, Rowley DA, Halliwell B, et al. Increased non-protein-bound iron and decreased protection against superoxide-radical damage in cerebrospinal fluid from patients with neuronal ceroid lipofuscinoses. Lancet 1982;ii:459-460.

234—Willson RL. Iron, zinc, free radicals and oxygen tissue disorders and cancer control. In: Iron Metabolism. Ciba Foundn Symp 51 (new series). Amsterdam: Elsevier; 1977:331-354.

235—Gutteridge JMC. Fate of oxygen free radicals in extracellular fluid. Biochem Soc Trans 1982;10:72-74.

236—Wills ED. Mechanisms of lipid peroxide formation in tissues: role of metals and haematin proteins in the catalysis of the oxidation of unsaturated fatty acids. Biochem Biophys Acta 1965;98:238-251.

237—Gutteridge JMC, Rowley DA, Halliwell B. Superoxide dependent formation of hydroxyl radicals and lipid peroxidation in the presence of iron salts. Biochem J 1982;206:605-609.

238—Heys AD, Dormandy TL. Lipid peroxidation in iron overloaded spleens. Clinical Science 1981;60:295-301.

238A—Cassinerio E, Roghi A, Pedrotti P, Brevi F, Zanaboni L, Graziadei G, Pattoneri P, Milazzo A, Cappellini MD. Cardiac iron removal and functional cardiac improvement by different iron chelation regimens in thalassemia major patients. Ann Hematol. 2012 May 10. [Epub ahead of print]

239—Ericson JE, Shirahata H, Patterson CC. Skeletal concentrations of lead in ancient Peruvians. N Engl J Med 1979;300:946-951.

240—Schroder HA. The Poisons Around Us. Bloomington: Indiana University Press; 1974:49.

241—Jenkins DW. Toxic Trace Metals in Mammalian Hair and Nails. US Environmental Protection Agency publication No.(EPA)-600/4-79049. Environmental Monitoring Systems Laboratory, 1979. (Available from National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161.)

242—Cranton EM, Bland JS, Chatt A, et al. Standardization and interpretation of human hair for elemental concentrations. J Holistic Med 1982;4:10-20.

243—Hansen JC, Christensen LB, Tarp U. Hair lead concentration in children with minimal cerebral dysfunction. Danish Med Bull 1980;27:259-262.

244—Medeiros DM, Pellum LK, Brown BJ. The association of selected hair minerals and anthropometric factors with blood pressure in a normotensive adult population. Nutr Research 1983;3:51-60.

245—Moser PB, Krebs NK, Blyler E. Zinc hair concentrations and estimated zinc intakes of functionally delayed normal sized and small-for-age children. Nutr Research 1982;2:585-590.

246—Thimaya S, Ganapathy SN. Selenium in human hair in relation to age, diet, pathological condition and serum levels. Sci Total Environ 1982;24:41-49.

247—Musa-Alzubaida L, Lombeck I, Kasperek K, et al. Hair selenium content during infancy and childhood. Eur J Pediatr 1982:139:295-296.

248—Gibson RS, Gage L. Changes in hair arsenic levels in breast and bottle fed infants during the first year of infancy. Sci Total Environ 1982;26:33-40.

249—Ely DL, Mostardi RA, Woebkenberg N, et al. Aerometric and hair trace metal content in learning-disabled children. Environ Res 1981;25(2):325-339.

250. —Yokel RA. Hair as an indicator of excessive aluminum exposure. Clin Chem 1982;28(4):662-665.

251—Bhat RK, et al. Trace elements in hair and environmental exposure. Sci Total Environ 1982;22(2):169-178.

252—Hurry VJ, Gibson RS. The zinc, copper, and manganese status of children with malabsorption syndromes and inborn errors of metabolism. Biol Trace Element Res 1982;4:157-173.

253—Thatcher RW, Lester ML, McAlester R, et al. Effects of low levels of cadmium and lead on cognitive functioning in children. Arch Environ Health 1982;37(3):159-166.

254—Peters HA, Croft WA, Woolson EA, et al. Arsenic, chromium, and copper poisoning from burning treated wood. N Engl J Med 1983:308(22):1360-1361.

255—Yamanaka S, Tanaka H, Nishimura M. Exposure of Japanese dental workers to mercury. Bull Toyko Den Coll 1982;23:15-24.

256—Capel ID, Spencer EP, Levitt HN, et al. Assessment of zinc status by the zinc tolerance test in various groups of patients. Clin Biochem 1982;15(2):257-260.

257—Vanderhoof JA, et al. Hair and plasma zinc levels following exclusion of biliopancreatic secretions from functioning gastrointestinal tract in humans. Dig Dis Sci 1983;28(4):300-305.

258—Foli MR, Henningan C, Errera J. A comparison of five toxic metals among rural and urban children. Environ Pollut Ser A Ecol Biol 1982;29:261-270.

259—Collipp PJ, Kuo B, Castro-Magana M, et al. Hair zinc levels in infants. Clin Pediatr 1983;22(7):512-513.

260—Medeiros DM, Borgman RF. Blood pressure in young adults as associated with dietary habits, body conformation, and hair element concentrations. Nutr Res 1982;2:455-466.

261—Huel G. Boudene C, Ibrahim MA. Cadmium and lead content of maternal and newborn hair: relationship to parity, birth weight, and hypertension. Arch Environ Health 1981;35(5):221-227.

262—Marlowe M, Folio R, Hall D, Errera J. Increased lead burdens and trace mineral status in mentally retarded children. J Spec Educ 1982;16:87-99.

263—Marlowe M, Errera J, Stellern J, et al. Lead and mercury levels in emotionally disturbed children. J Orthomol Psychiatr 1983;12(4):260-267.

264—Nolan KR. Copper toxicity syndrome. J Orthomol Psychiatr 1983;12(4):270-282.

265—Klevay L. Hair as a biopsy material-assessment of copper nutriture. Am J Clin Nutr 1970;23(8):1194-1202.

266—Rees EL. Aluminum poisoning in Papua New Guinea natives as shown by hair testing. J Orthomol Psychiatr 1983;12(4):312-313.

267—Cook JD, Finch CA, Smith NJ. Evaluation of the iron status of a population. Blood 1976;48:449-455.

267A—Sung KC, Kang SM, Cho EJ, Park JB, Wild SH, Byrne CD. Ferritin Is Independently Associated With the Presence of Coronary Artery Calcium In 12,033 Men. Arterioscler Thromb Vasc Biol. 2012 Jul 26. [Epub ahead of print]

268—Kannel WB, Hjortland MC, McNamara PM, et al. Menopause and the risk of cardiovascular disease. The Framingham Study. Ann Intern Med 1976;85:447-452

269—Hjortland MC, McNamara PM, Kannel WB. Some atherogenic concomitants of menopause: The Framingham Study. Am J Emidemiol 1976;103:304-311.

270—Gordon T, Kannel WB, Hjortland MC, et al. Menopause and coronary heart disease. The Framingham Study. Ann Intern Med 1978;89:157-161.

271—Sullivan JL. Iron and the sex difference in heart disease risk. Lancet 1981; 1(8233):1293-1294.

272—Casale G, Bignamini M, de Nicola P. Does blood donation prolong life expectancy? Vox Sang 1983;45:398-399.

273—Skoog DA, West DM. Volumetric methods based on complex-formation reactions. In: Fundamentals of Analytical Chemistry. New York: Holt, Rinehart and Winston, Inc; 1969:338-600.

273A—Cranton E M (ed): A Textbook on EDTA Chelation Therapy, Second Edition, Charlottesville, VA, Hampton Roads Publishing Co, 2001, Foreword by Linus Pauling, PhD. Chapter 37.

274—Peng CF, Kane JJ, Murphy ML, et al. Abnormal mitochondrial oxidative phosphorylation of ischemic myocardium reversed by calcium chelating agents. J Mol Cell Cardiol 1977;9:897-908.

275—Freeman AP, Giles RW, Berdoukas VA, et al. Early left ventricular dysfunction and chelation therapy in thalassemia major. Ann Intern Med 1983;99:450-454.

276—Blake DR, Hall ND, Bacon PA, et al. Effect of a specific iron chelating agent on animal models of inflammation. Ann Rheum Dis 1983;42:89-93.

277—Addonizo VP, Wetstein L, Fisher CA, et al. Medication of cardiac ischemia by thromboxanes released from human platelets. Surgery 1982;92:292.

278—Yagi K, Ohkawa H, Ohishi N, et al. Lesion of aortic intima caused by intravenous administration of linoleic acid hyperoxide. J Appl Biochem 1981;3:58-65.

279—Benditt EP. The origin of atherosclerosis. Scientific American. 1977 Feb:74-85.

280—McCullach KG. Revised concepts of atherogenesis. Cleue Clin Q 1976;43:247.

281—Mayron LW. Portals of entry—a review. Ann Allerg 1978;40:399-405.

282—Walker WA, Isselbacher KJ. Uptake and transport of macromolecules by the intestine: possible role in clinical disorders. Gastroenterology 1974;67:531-550.

283—Hemmings WA, Williams EW. Transport of large breakdown product of dietary protein through the gut wall. Gut 1978;19:715-723.

284—Rowe AH, Rowe AH Jr. Food Allergy: Its Manifestations and Control and the Elimination Diets. Springfield, Il: Charles C. Thomas; 1972.

285—Speer F, ed. Allergy of the Nervous System. Springfield, Il: Charles C. Thomas; 1970.

286—Dickey LD, ed. Clinical Ecology. Springfield, Il: Charles C. Thomas; 1976.

287—Randolph TG. Human Ecology and Susceptibility to the Chemical Environment. Springfield Il: Charles C. Thomas; 1962.

288—Crook WG. The coming revolution in medicine. J Tenn Med Assn 1983;76(3):145-149.

289—Iwata K. Toxins produced by Candida albicans. Contr Microbiology Immunol 1977;4:77-85.

290—Iwata K, Yamamota Y. Glycoprotein Toxins Produced by Candida albicans. Reprinted from Proceedings of the Fourth International Conference on the Mycoses. PAHO Scientific Publication 1977;356:246-257.

291—Iwata K. Fungal toxins as a parasitic factor responsible for the establishment of fungal infections. Mycopathologia 65:141-154.

292—Crook WG. The Yeast Connection: A Medical Breakthrough. Jackson, TN: Professional Books; 1983.

293—Truss CO. Tissue injury induced by Candida albicans. Orthomolecular Psychiatry 1978;7(1):17-37.

294—Truss CO. Restoration of immunologic competence to Candida albicans. Orthomolecular Psychiatry 1980;9(4):287-301.

295—Truss CO: The role of Candida albicans in human illness. Orthomolecular Psychiatry 1981;10(4):228-238.

296— Hatano S, Nishi Y, Usui T. Copper levels in plasma and erythrocytes in healthy Japanese children and adults. Am J Clin Nutr 1982;35:120-126.

297— Harrison W, Yarachek J, Benson C. The determination of trace elements in human hair by atomic absorption spectroscopy. Clin Chem Acta 1969;23(1):83-91.

298— Gori GB. Observed no-effect thresholds and the definition of less hazardous cigarettes. Journal of Environmental Pathology and Toxicology 1980;3:193-203.

299— Saltin B, Karlsson J. Muscle Metabolism During Exercise. New York: Plenum Publishing Co; 1971:395.

300— Frustaci A, Magnavita N, Chimenti C, Caldarulo M, Sabbioni E, Pietra R, Cellini C, Possati GF, Maseri A. Marked elevation of myocardial trace elements in idiopathic dilated cardiomyopathy compared with secondary cardiac dysfunction. J Am Coll Cardiol. 1999 May;33(6):1578-83.

301— KJ, Volitakis I, Fraser FW, Kim Y, Huang X, Goldstein LE, Moir RD, Lim JT, Beyreuther K, Zheng H, Tanzi RE, Masters CL, Bush AI. Treatment with a copper-zinc chelator markedly and rapidly inhibits beta-amyloid accumulation in Alzheimer's disease transgenic mice. Neuron. 2001 Jun;30(3):665-76.

302— Belland RJ, Ouellette SP, Gieffers J, Byrne GI.Chlamydia pneumoniae and atherosclerosis.. Cell Microbiol. 2004 Feb;6(2):117-27.

303— Mussa FF, Chai H, Wang X, Yao Q, Lumsden AB, Chen C. Chlamydia pneumoniae and vascular disease: an update. J Vasc Surg. 2006 Jun;43(6):1301-7.

304— Dolan RM, Costerton JW. Biofilms: Survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 202;15:167-93.

Copyright © 2013 Elmer M. Cranton, M.D., all rights reserved

Last modified: