What causes Free radicals?

There are many internal and external factors which form free radicals: tobacco smoke, excessive alcohol, radiation including ultraviolet radiation from the sun, auto exhaust, pesticides, herbicides, pollution, prescription drug use, chemotherapy, surgery, breakdown of bacteria by white blood cells, microbial or viral infections, metabolism of toxins, inflammatory processes, byproducts of oxygen metabolism, stress, shock, trauma, hypoxia, enzymatic reactions, calorie consumption, poor diet and many food materials especially oxidizing hydrogenated oils.

A single free radical can destroy an enzyme, a protein molecule, a strand of DNA, or an entire cell, but even worse, in a nano-second it can unleash torrential chain reactions in our bodies. Each free radical can initiate and perpetuate millions of other free radicals, setting off chains of biologically damaging reactions. This damage is at the molecular and cellular levels. Ironically, the underlying mechanisms that most chemotherapeutic agents and ionizing radiation have is not to neutralize free radicals but to produce more free radicals which leads to irreversible tissue injury.

What Are Antioxidants?

In the most simplistic terms, the role of antioxidants is to interact with free radicals and "quench" them or render them harmless. Researchers believe that increased dietary intake of antioxidants can slow the process of free radical damage and associated disorders. By removing free radicals, antioxidants help to: protect against DNA damage in cells, protect cell membranes, protect against all forms of cancer, protect the brain against various forms of dementia, protect against the harmful cross-linking of proteins with sugars that cause cell damage and may help slow the aging process. Antioxidants have been shown to provide: blood vessel strength and protection, enhanced memory and learning function, healthy lung function, bone and joint flexibility.

According to the National Cancer Institute, "Antioxidants are substances that may protect cells from the damage caused by unstable molecules known as free radicals. Free radical damage may lead to cancer. The antioxidants interact with and stabilize free radicals and may prevent some of the damage free radicals otherwise might cause. Considerable laboratory evidence from chemical, cell culture, and animal studies indicate that antioxidants may slow or possibly prevent the development of cancer. Antioxidants are often described as "mopping up" free radicals, meaning they neutralize the electrical charge and prevent the free radical from taking electrons from other molecules."

US Federal Courts have recently instructed the FDA to allow the claim that antioxidants may prevent cancer (Whitaker vs. Thompson {2002}, Pearson vs. Shalala {1999} and Pearson vs. Shalala 2 {2001}). These landmark rulings benefit the general public, which now has access to this extremely important health information that may help millions of people reduce the risk of this dreaded disease and live longer as a result of using antioxidants in their diet.

Two principal mechanisms of action have been proposed for antioxidants, first is stabilizing free radical present in the system, and the second mechanism involves the removal of chain initiating catalysts. Furthermore, antioxidants can act by scavenging biologically important reactive oxygen species, by preventing their formation, or by repairing the damage that they do. Antioxidants can suppress apoptosis (programmed cell death), act as reducing agents, chelate metal compounds and affect directly or indirectly the expression of genes in tissues. A diet high in antioxidants may even bolster the body's own defenses against biological invaders transmitted by germ warfare, mosquitoes or other delivery methods.

The body's arsenal of antioxidants appear to be sufficient for keeping oxidation in check in children and in youths, but once we reach our 20's, the effectiveness of the body's antioxidants defense mechanisms lessen and free radicals are given greater rein to do damage. For example, the antioxidant enzyme, superoxide dismutase, appears to diminish with age and the antioxidant capacity in human plasma decreases. While the body's antioxidant defenses are reduced, the number of free radicals in the body rises dramatically. Studies also show that cells from old individuals are more susceptible to oxidative damage than cells from younger donors and that some of this damage can actually be prevented by antioxidants.

Researchers found that cancer patients with small cell lung cancer who used antioxidants showed increased long-term survival rates as compared with previously published studies. It was also noticed that patients receiving antioxidants were able to tolerate chemotherapy and radiation treatment well. It was concluded that antioxidant treatment could potentiate orthodox cancer treatments by decreasing the likelihood of side effects and increasing the host immune defense. Ralph Moss, PhD, author of the best selling book, "The Cancer Industry," former assistant director of public affairs at Memorial Sloan-Kettering Cancer Center, and producer for several documentaries, including, "The Cancer War," stated in his newsletter of 8/17/2003; "I would argue that the preponderance of evidence already suggests that antioxidants reduce the side effects of chemotherapy and radiation, without, however, interfering with their effectiveness.

As the body's own antioxidant defenses are gradually overwhelmed by the aging process or disease or both, fruits, vegetables, herbs, spices, and concentrated dietary supplements should be consumed as they contain the largest amounts of antioxidants to help replenish and augment the system.

Various Forms of Free Radicals and Antioxidants

There are 6 common reactive species of free radicals existing in the body. Radicals of oxygen compromise the variety of reactive molecules that can constitute oxidative stress to the cells. The reactive oxygen species are the peroxyl radical, hydroxyl radical and superoxide ion. Nitric oxide, singlet oxygen and peroxynitrite are the other reactive forms of radicals.

As free radicals are all different, likewise, all antioxidants are not alike. Not only do antioxidants differ in their potency but also on what type of free radicals they work upon. For example, Vitamin C only works on water-soluble peroxyl radicals, vitamin E scavenges fat-soluble peroxyls, superoxide dismutase and catalase is only effective on superoxide radicals, etc. It is rare to find an antioxidant that can work effectively on more than one type of radical let alone all 3 forms of the harmful reactive oxygen species. To properly understand the significance of our independent study, a brief discussion of the three reactive oxygen species that Cantron was tested on is necessary.

The Peroxyl Radical

Free radicals attack all major classes of bio-molecules, but lipids are the most susceptible and the easiest to damage. The peroxyl radical species is reasonably stable and not very reactive but they are reactive enough, however, to attack adjacent fatty acid side chains, as well as enzymes, receptors, and other structures found in cell membranes. The cell membrane is a rich source of polyunsaturated fatty acids, which are easily attacked by oxidizing radicals including the peroxyl radical. The destruction of polyunsaturated fats causes damage by unleashing a chain reaction of chemical events that can collapse cell membranes. Once one peroxyl free radical forms, and an appropriate antioxidant is not available to stop the process, the chain of events keeps occurring until the cell membrane literally collapses. As the cellular membrane becomes compromised, the cell bursts open, spews its contents and dies. This series of damaging breaks in the cellular membrane can be prevented by antioxidant defenses and the cells can remain intact. However, it has been demonstrated that total peroxyl radical scavenging antioxidant capacity (TRAP) in human plasma decreases with age.

Peroxyls are formed by several routes especially during the breakdown of organic peroxides, oxidation of lipids or other organic molecules in oxidative stress. They are formed within the delicate cellular membrane. If a free radical within or on the outside of a cell attacks the fatty acid cell membrane structures, they create peroxyl free radicals. These radicals are also formed as a byproduct of the clash between hydroxyl radicals and polyunsaturated fatty acids derived from vegetable oils.

The Hydroxyl Radical

These dangerous radicals can attack and damage almost every molecule found in living cells because they react as soon as they come in contact with another molecule. Since it is so reactive, hydroxyl radicals generated in vivo do not persist for even a microsecond as they rapidly combine with molecules in their immediate vicinity as fast as they collide. Hydroxyl radicals can be produced at an enormous rate, have easy access to every portion of the cell, are capable of causing great damage within a small radius of their site of production and are highly carcinogenic. In addition to damaging unsaturated fats in cell membranes, hydroxyls are reactive enough, aggressive enough, and persist long enough to damage the less susceptible proteins (including the fragmentation of vital proteins in plasma), nuclear acids, enzymes and carbohydrates.

Russell Reiter, PhD, professor of neuroendocrinology at University of Texas Health Center has highlighted the dangers of the hydroxyl radical; "If the function of radicals is to destroy molecules and tissues, then the hydroxyl radical would be the radical’s radical. It reacts at diffusion rates with virtually any molecule found in its path including macromolecules such as DNA, membrane lipids, proteins and carbohydrates. In terms of DNA, the hydroxyl radical can induce strand breaks as well as chemical changes in the dioxyribose and in the purine and pyrirnidine bases. Damaged proteins, many of them crucial enzymes in neurons lose their efficiency and cellular function wanes. Protein oxidation in many tissues, including the brain, has been proposed as an explanation for the functional deficits associating with aging."

In addition to the direct damage caused by hydroxyls, they play a major role in forming peroxyl radicals and stimulating the free radical chain reaction known as lipid peroxidation. Peroxyl radicals are formed when oxygen combines with the hydrogen radical. One hydroxyl radical can result in the conversion of many hundred fatty acids side chains into lipid hydroperoxides. As hydroxyl radicals react with carbohydrates it leads to chain breaks in important molecules in a process involving the peroxyl radical as an intermediate. Since hydroxyls do such direct damage, work in conjunction with and create harmful peroxyl radicals, then supplementation of hydroxyl scavenging antioxidants may be extremely important.

The Superoxide Radical

The body utilizes important antioxidants to deal with superoxide radicals, in particular the enzyme Superoxide Dismustase (SOD). Decreased SOD favors Superoxide anion formation. This antioxidant is so necessary that its very absence would be lethal. The problem is that SOD levels in the body decline with age and supplementation with SOD tablets is not that effective because they are poorly absorbed into the bloodstream.

To Recap

• It is a well-established fact that free radicals have been implicated in over 50 degenerative diseases including heart disease, cancer and aging.
• Consumption of antioxidants may eradicate dangerous free radicals and help prevent disease.
• The federal court has forced the FDA to allow the claim that antioxidants may prevent cancer.
• The body’s storehouse of antioxidants diminish with age and leads to the onset of disease and premature aging.
• There are many forms of free radicals. Peroxyl radicals are the most abundant, hydroxyl radicals are the most dangerous and superoxide anions are the greatest initiator of other dangerous free radicals.
• One free radical engenders many other free radicals, causing a chain reaction of biological events.
• Cantron is not only the most potent antioxidant known to man, but works against a number of radicals that are responsible for multitudes of diseases in men.
• To obtain the equivalent antioxidant protection of just one daily dose of Cantron (1 1/2 teaspoons), one would have to consume 44 ounces of orange juice (5 1/2 cups) for it peroxyl scavenging activities, 26 teaspoons of concentrated green tea extract with 95% polyphenols for its hydroxyl radical scavenging ability, and mega-doses of superoxide dismutase and/or catalase tablets.
• Supplementation of antioxidants, along with a healthy diet of antioxidant foods and beverages should be exercised by anyone who values good health or who is seeking “Wellness.” A wide variety of antioxidant substances are recommended in our “Cantron Total Wellness Program” and our “Cantron Total Wellness Diet.”

Cantron has been known to be a powerful antioxidant. Research was needed, however, to determine just how potent Cantron is in relation to other powerful antioxidants and to ascertain which types of Reactive Oxygen Species (ROS) it works upon. Most antioxidants usually scavenge only one specific type of radical species, for example, vitamin C only works upon water-soluble peroxyl radicals and has no effect upon fat-soluble peroxyl radicals, hydroxyl radicals, or superoxide radicals.

Nutraceutical chemistry consultant, Daniel Hetrick, PhD., was commissioned to design, implement and oversee this project. It was decided to test Cantron on all 3 of the reactive oxygen species that wreak havoc on the body and cause severe illnesses. It was also decided to test previous generations and other variations of the Cantron formula including: Sheridan’s Entelev, Sopcak’s Cancell, and 3 different variations of the Cantron formula (our original Sopcak/Feather version, our current New Millennium Version and an experimental version). All versions of this formula have the same basic ingredients - the only difference being the quantities and ratios of those components that are created in the manufacturing process. The results of the different variations were not expected to vary widely, if at all. Cantron was also tested for its phenolic content. Phenolic and polyphenolic compounds are known as powerful antioxidant substances that exist in many plant foods and herbs.

Samples were sent to Brunswick Laboratories in Wareham, MA (www.brunswicklabs.com) to test for phenolic content and activity against peroxyl and hydroxyl radicals. To test for activity of superoxide radicals, samples were sent to Cayman laboratories in Ann Arbor, MI (www.caymanchem.com).

Oxygen Radical Absorption Capacity and Hydroxyl Radical Absorption Capacity of Cantron and Related Formulas

The oxygen radical absorption capacity test (ORAC) and Hydroxyl Radical Absorption Capacity test (HORAC) was carried out at Brunswick Laboratories, a state of the art facility with highly skilled scientists. Brunswick serves the nutraceutical, pharmaceutical, food and cosmetic industries. Their antioxidant team developed proprietary high throughput technologies in antioxidant screening and has accumulated the world’s largest antioxidant database on a wide variety of natural product compounds. Their expertise has made them a leader in antioxidant activity profiles. Brunswick Labs is the inventor of the improved and automated ORAC assay (US patent Pending). This assay is the second generation of the ones used by the National Institute of Health and US Department of Agriculture and is more accurate than its predecessors. Brunswick screens natural products, antioxidant supplements, beverages, pure chemicals, plasma, serum, and urine in their antioxidant assays.

The ORAC test measures the antioxidant activity against peroxyl radical by determining how many peroxyl free radicals can be absorbed by a given antioxidant as compared to a vitamin E analog (Trolox) as the test standard and vitamin C. Since Cantron is an aqueous product, the test radicals utilized in this study were hydrophillic peroxyl radicals (note: vitamin E is lipophillic). The original ORAC assay was primarily for water-soluble antioxidant activity against peroxyl radicals. Recently Brunswick Laboratories expanded the ORAC method to incorporate fat-soluble antioxidant activity.

The HORAC measures the activity of a given antioxidant against hydroxyl radicals. Cantron and related formulations were to be tested and to be compared to the test standard, Gallic Acid, one of the most common and powerful antioxidants found in green vegetables such as broccoli and spinach. Since Cantron is an aqueous product, the test radicals utilized in this study were hydrophillic hydroxyl radicals.

In February 2003, two samples of the Cantron formula were sent to Brunswick Laboratories: the current New Millennium version and an experimental version. Brunswick was to test the Cantron samples and to send the raw scores expressed in units to Dr. Hetrick who in turn was to make the necessary calculations comparing Cantron to the various test standards.

For the ORAC test, Trolox, a water-soluble Vitamin E analog, is used as the calibration standard and the ORAC unit is expressed as micromole Trolox equivalent (TE) per liter (uMole TE/L). The ORAC value for the vitamin E standard is 1200 units (1200 uMole TE/L) and the value for the vitamin C standard is 5000 units (5,000 uMole TE/L). Because Cantron is a liquid formulation we also decided to compare its ORAC values to that of an equivalent amount of orange juice, which contains vitamin C, bioflavonoids and other antioxidant factors. Orange juice has a value of 7,400 units, which is slightly higher than the vitamin C standard. For the HORAC test, Gallic Acid is used as the calibration standard and the HORAC result is expressed as mg. Gallic Acid equivalent (GAE) per liter. Gallic Acid is a potent antioxidant that exists in vegetables such as broccoli. The HORAC value for Gallic acid is 1,000 units (1,000mgGAE/gL).

Because each variation of Cantron has a slightly different density, in order to make a proper comparison, the raw scores of the study needed to be adjusted for the density of the sample, before illustrating the comparative values. The results of these 2 samples came back on February 19, 2003 (copies of the report are available upon request). The phenolic content of Cantron was measured at an incredibly high 86,659.24 mg/L and the results of the Cantron formula against peroxyl and hydroxyl radicals were astonishing based upon the values of other potent antioxidant substances that have previously been tested throughout the years.

The ORAC score for the New Millennium version of Cantron - after adjusting for density - was a whopping 1,081,886 units as opposed to a mere1200 for vitamin E and 5,000 for vitamin C. Comparatively speaking, this version was 902 times more powerful than the vitamin E standard, 216 times more powerful than vitamin C standard and 142 times more powerful than orange juice against peroxyl radicals. The experimental version was even more potent against peroxyl radicals. It scored 1,135,591 making it 942 times more effective than vitamin E, 226 times vitamin C and 153 times orange juice. Surprisingly, however, while the experimental formula worked better against peroxyl radicals, the New Millennium version was far superior on hydroxyl radicals, scoring 45,003 (45 times Gallic Acid) to 33,180 (33 times Gallic Acid).

Samples of 3 other variations of the formula were sent to Brunswick Labs in April 2003. These samples included: a bottle of Entelev that was manufactured by the original developer, James V. Sheridan, in 1982, a bottle of Cancell that was manufactured by Edward Sopcak in 1988, and a bottle of 1997 vintage Cantron manufactured by Medical Research Products. This 1997 version of Cantron utilized the same manufacturing procedures as Ed Sopcak did in his Cancell formulation and was chemically identical; therefore, results were expected to be identical or closely similar. MRP manufactured and distributed this version from 1984 to 1998 before the new, improved version was released. Andy Johnson a member of the Hope Foundation provided the bottles of Entelev and Cancell.

On May 1, 2003, Brunswick completed their report (copies available upon request). Unexpectedly, the results of these samples were dramatically different than the first two samples. The 1982 Entelev formula scored 879,050 in the ORAC test, the 1997Cantron scored 2,122,527, and the 1988 Cancell product scored 1,882,618. The 97 Cantron and 88 Cancell formulas worked significantly better on peroxyl radicals than the first samples, but rather surprisingly; they had almost no effect on hydroxyl radicals - the most dangerous radical of all. None of these 3 formulas were as effective as the Gallic Acid standard of 1,000 units. Entelev scored 161, 1997Cantron scored 236 and 1988 Cancell scored 215 as compared to New Millennium Cantron’s 45,003.

Because Cantron is in liquid form - not in pill form as contrasted to the established test standards of vitamin C and E in the ORAC study - the comparison of Cantron to liquid orange juice has been included in this report. Orange juice, which has an ORAC value of 7,400 (slightly higher than the vitamin C standard) contains vitamin C, bioflavonoids and other antioxidant substances. The problem with this comparison, however, is related to dosage/consumption. Although an equivalent volume of New Millennium Cantron is 142 times more effective than orange juice, the daily dosage for Cantron is small (11/2 teaspoons). Obviously, one is able to consume a great deal more orange juice than Cantron. But the small daily dose of Cantron still provides tremendous antioxidant benefits. To obtain equivalent peroxyl scavenging protection of a daily dose of Cantron, one would have to consume 44 ounces of orange juice (5 1/2 cups). And since orange juice is not effective upon hydroxyl or superoxide radicals, to get the equivalent protection of a daily dose of Cantron one would also have to consume 11/2 cups (12 oz.) of green tea that contains at least 95% polyphenols and mega doses of superoxide dismutase and/or catalase. Naturally, one could further increase their antioxidant protection by taking the above-mentioned antioxidants or other potent antioxidants along with Cantron.

Brunswick Laboratories has compiled a vast and comprehensive antioxidant database. Green Tea was by far the greatest hydroxyl radical scavenger on their chart; in fact, it is 9.2 times more potent than the next highest antioxidant (eldeberry extract). Volume for volume, Cantron is 16 times more powerful than Green Tea (147 times eldeberry extract), which demonstrates its vast superiority to all other hydroxyl radical scavengers. Superoxide Dismutase and Catalase do not absorb well, and data is not available at this writing that could quantify the actual intake of these substances, which that would be equivalent to one dose of Cantron, but it would certainly take a significant amount. When data is available, it will be reported on Cantron.com.

The extraordinarily high phenolic content of Cantron (86659.24 mg/L) is ascribed to the presence of a series of quinones, which cross-react in the phenol assay. Cantron does not contain free phenol.

Conclusion

Cantron has an extremely high phenolic content and all variations of the formula have extremely high antioxidant activity. Some of the formulas were more active on peroxyl radicals, while others were more effective against hydroxyl radicals. Cantron New Millennium formula had the most balanced antioxidant activity, having substantial activity against both hydroxyl and peroxyl radicals.

HORAC Scores

ORAC Scores

ORAC Values Of Various Antioxidant Juices & Liquid Cantron    (ORAC Value Per Liter)

Cantron: An Assay of Superoxide Radical Scavenging Ability

Scavenging of superoxide radicals cannot be directly quantified. Instead, measurements of the activity of the superoxide specific antioxidant enzyme, superoxide dismutase (SOD), are taken after samples of a given antioxidant are added to a solution of superoxide anions.

The Cayman Laboratories assay for SOD activity utilizes superoxide anions generated from Xanthine Oxidase. The anion production is used to measure the activity/concentration of SOD. The assay is modified by inserting the test samples- in this case Cantron and related formulations- after the generation of superoxide anions, but before the addition of SOD. Thus if the Cantron samples inactivate the superoxide anion, activity of SOD will be less than the untreated controls. The reduction in SOD activity/concentration is expressed in units/ml and is the measure of Cantron’s ability to destroy the superoxide anions.

Cayman assayed the samples of Cantron at solutions ranging from 1:1 to 1:16,000. At the lower dilutions, the opacity of the samples often interfered with the absorbency measurements and blocked the readings. The first readable sample was at a dilution of 1:10 (diluted 10 times). At this dilution, all of the Cantron samples tested were able to scavenge all of the generated superoxide anions. The endpoint dilution for activity of the samples lies somewhere between 1:10 and 1:100. All dilutions beyond that had no inhibitory effect. The control value at the dilution of 1:10 was + .129 units/ml. Therefore, any score less than this figure would result in a decrease in SOD activity. A zero or negative number indicates no SOD activity whatsoever, and conversely, complete blockage of Cantron against superoxide radicals. All formulas received a negative score.

Conclusion

At a dilution between 1:10 to 1:100, all samples tested had high activity as superoxide anion scavengers or inactivators. At dilutions of 1:10, all samples exhibited complete scavenging ability against superoxide radicals.

Table: Results for Superoxide Anion Scavenging in the SOD Assay (Cayman Laboratories)

Discussion of the Two Studies

The results of the scientific studies conducted at both laboratories indicate that Cantron is not only the most potent but also perhaps the most versatile antioxidant ever tested. In addition to being highly effective upon all three forms of reactive oxygen species, the other damaging radical species such as peroxynitrite, hydrogen peroxide and singlet oxygen are also averted because the propagating radicals (superoxide and hydroxyl) are quenched before these other radicals can be formed.

The most puzzling question is why the various formulations differ so widely in their ORAC and HORAC scores. As the components are the same and the ratios of the ingredients are not that different, results were expected to be similar or the same. With only one exception (1982 Entelev), an inverse relationship or “seesaw” effect was manifested, i.e., whenever the HORAC went up, the ORAC simultaneously went down and vice versa. The question is; are the various formulas actually more or less effective against certain radicals or is there another scientific explanation? Hydroxyl radicals are known to initiate the formation of peroxyl radicals and peroxyls are known to play an intermediary role with hydroxyls, causing a wide range of chain reactions. Could it be that this complex relationship between the two radicals is responsible for changing the research results so dramatically? Could it be that the prevention or interference of certain chain reactions are responsible? These questions will be addressed in future investigations?

As expected, because the 1997 Cantron and the 1988 Cancell utilized the same manufacturing procedure, their performance was virtually the same, albeit, 97 Cantron was slightly more effective. The scores are even closer before the adjustment for density was made. These two samples had the highest ORAC scores (Cantron 1,798,752 to Cancell’s 1,727,173), among the lowest HORAC scores (Cantron 205, Cancell 187), and were equally effective against superoxide anions (Cantron -.66, Cancell -.67). The similar behavior of these two samples certainly proves that they are virtually identical, with 97 Cantron being slightly superior in its scavenging abilities to the Cancell.

It would be hard to dispute that New Millennium Cantron was the most effective and most complete antioxidant formula in this group as it had the highest hydroxyl radical absorption capacity along with a very high-capacity to scavenge peroxyl radicals. Hydroxyl radicals are the most dangerous radicals known to chemistry and are also known to initiate peroxyl radicals.

Final Conclusion of the Two Studies

Cantron contains a high level of phenolic compounds. Among all formulas tested, New Millennium Cantron has the most balanced antioxidant effect, as it is highly effective upon all three forms of the Reactive Oxygen Species of free radicals. By quenching the initiating radicals it may also help prevent the formation of other dangerous radicals such as peroxynitrite, hydrogen peroxide and singlet oxygen. Future tests on in vivo activity of Cantron are warranted.

The FDA has not evaluated the statements in this article or the research results.

For Further Laboratory Studies on Other Variations of Cantron
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