Ozone, an allotropic form of oxygen, possesses unique properties which are being defined and applied to biological systems as well as to clinical practice. As a molecule containing a large excess of energy, ozone, through incompletely understood mechanisms, manifests bactericidal, virucidal, and fungicidal actions which may make it a treatment of choice in certain conditions and an adjunct to treatment in others.

The oxygen atom exists in nature in several forms: (1) As a free atomic particle (0), it is highly reactive and unstable. (2) Oxygen (02), its most common and stable form, is colorless as a gas and pale blue as a liquid. (3) Ozone (03), has a molecular weight of 48, a density one and a half times that of oxygen, and contains a large excess of energy in its molecule (03 -> 3/2 02 + 143KJ/mole). It has a bond angle of 127 ± 3, resonates among several forms, is distinctly blue as a gas, and dark blue as a solid. (4) 04 is a very unstable, rare, nonmagnetic pale blue gas, which readily breaks down into two molecules of oxygen.

Ozone is a powerful oxidant, surpassed in this regard only by fluorine. Exposing ozone to organic molecules containing double or triple bonds yields many complex and as yet incompletely configurated transitional compounds (i.e. zwitterions, molozonides, cyclic ozonides), which may be hydrolysed, oxidized, reduced, or thermally decomposed to a variety of substances, chiefly aldehydes, ketones, acids or alcohols. Ozone also reacts with saturated hydrocarbons, amines, sulfhydryl groups, and aromatic compounds.

Importantly relevant to biological systems is ozone's interaction with tissue--including blood--constituents. The most studied is lipid peroxidation, although interactions have yet to be more fully investigated with complex carbohydrates, proteins, glycoproteins, and sphingolipids.

Ozone is a pan-virucidal, and a pan-bactericidal agent. In addition, it is well documented that many species of fungi are inactivated by its actions, as well as several types of protozoa.

Ozone is a gas which, properly interfaced with biological systems or pathologically afflicted tissues, exerts significant therapeutic activity. As is the case with many medications, however, ozone has a range of therapeutic action which, in the terminology of pharmacokinetics, is termed a therapeutic window. Indeed, ozone applied in concentrations that are too low, has little therapeutic effect. More importantly, when it is applied in too high concentrations, it is known to have some toxic sequelae.

Due to ozone's demarcated therapeutic range, ozone concentrations administered to the patient need to be carefully calibrated and controlled. The therapeutic ozone/oxygen mixture requires state of the art quantitative (dosage, concentration), as well as qualitative (purity) controls, which can only be provided by an appropriate contemporary technology.

At room temperature, approximately 50% of ozone reverts to pure oxygen. This adds an important dimension to the calculation of the amount of ozone administered. As regards the generation and delivery system, of foremost importance is the oxygen source which must be of medical grade purity, and thus devoid of nitrogen or impurities. The presence of nitrogen favors the production of nitrogen oxides which are tissue-toxic. Due to these considerations, ozone needs to be conceptualized as a medication with complex therapeutic dynamics, which need to be carefully considered and evaluated in relation to the particular medical conditions being treated.

Ozone generation and delivery systems are intrinsically connected to the fact that ozone, utilized for human or veterinary therapeutic purposes, requires that it be created at the moment it is to be administered. Ozone, in this sense is not a drug that has a shelf life, and that can be kept for long periods of time at a certain determined dosage. As a gas with a half life of approximately one hour at room temperatures, the gauging of ozone's dosage is intrinsically connected to the sophistication of its manufacture technology and its pharmacodynamics.

Ideally, the treating clinician should be able to be informed of the exact concentration of the ozone drug being generated and delivered (i.e., a digital readout of ozone output in micrograms per milliliter, or grams per cubic meter). In addition, the clinician needs to factor the natural and constant conversion of ozone into oxygen, so as to arrive at precise measurements of dosage in relation to duration of administration.

In the case of external application, the ozone generator supplies a dosage of ozone/oxygen determined by the clinician to be therapeutically indicated. This, in practice, may involve an infected foot, a post-surgical incision, an area afflicted by a burn, a decubitus ulcer, or a poorly healing post-traumatic wound.

In the practice of external ozone application, a specially designed polyester envelope is used to enclose the area under treatment. A precise fitting of the bag is needed in order to ensure (I) A proper constant concentration of delivered ozone, (2) A suitable containment of ozone/oxygen to the affected area. This guarantees that ozone will be prevented from escaping into the ambient environment which, in higher concentrations, may lead to respiratory epithelial irritation in the patient or in the treating personnel, and (3) An opportunity for the precise timing of the duration of ozone exposure under controlled conditions.

In order to respect proper environmental controls, and to prevent ozone from diffusing into the treatment space, an exit catheter connected to the polyethelene envelope is directed to the ozone generator for catalytic reconversion to oxygen.

Externally applied ozone concentrations need to be carefully adjusted. The clinician must be able to gauge the proper ozone concentration geared to the specific medical condition under treatment. In wet burns, for example, initial ozone concentrations will need to be low, in order to prevent inordinate systemic absorption. As the burn heals, and progressively dries, greater ozone concentrations may then be administered in order to keep pace with the rate of healing.

The antipathogenic effects of ozone have been substantiated for several decades. Its killing action upon bacteria, viruses, fungi, and in many species of protozoa, serve as the basis for its increasing use in disinfecting municipal water supplies in cities worldwide.

Indicator bacteria in effluents, namely coliforms and pathogens such as Salmonella, show marked sensitivity to ozone inactivation. Other bacterial organisms susceptible to ozone's disinfecting properties include Streptococci, Shigella, Legionella pneumophila, Pseudomonas aeruginosa, Yersinia enterocolitica, Campylobacter jejuni, Mycobacteria, Klebsiella pneumonia, and Escherichia coli. Ozone destroys both aerobic, and importantly, anaerobic bacteria which are mostly responsible for the devastating sequelae of complicated infections, as exemplified by decubitus ulcers and gangrene.

The mechanisms of ozone bacterial destruction need to be further elucidated. It is known that the cell envelopes of bacteria are made of polysaccharides and proteins, and that in Gram negative organisms, fatty acid alkyl chains and helical lipoproteins are present. In acid-fast bacteria, such as Mycobacterium tuberculosis, one third to one half of the capsule is formed of complex lipids (esterified mycolic acid, in addition to normal fatty acids), and glycolipids (sulfolipids, lipopolysaccharides, mycosides, trehalose mycolates). The high lipid content of the cell walls of these ubiquitous bacteria may explain their sensitivity, and eventual demise, subsequent to ozone exposure. Ozone may also penetrate the cellular envelope, directly affecting cytoplasmic integrity, disrupting any one of numerous levels of its metabolic complexities.

Numerous families of viruses including poliovirus I and 2, human rotaviruses, Norwalk virus, Parvoviruses, and Hepatitis A, B, and non-A non-B (C), among many others, are susceptible to the virucidal actions of ozone.

Most research efforts on ozone's virucidal effects have centered upon ozone's propensity to break apart lipid molecules at sites of multiple bond configuration. Indeed, once the lipid envelope of the virus is fragmented, its DNA or RNA core cannot survive.

Non-enveloped viruses (Adenoviridae, Picornaviridae, namely poliovirus, Coxsachie, Echovirus, Rhinovirus, Hepatitis A and E, and Reoviridae (Rotavirus), have also begun to be studied. Viruses that do not have an envelope are called "naked viruses." They are constituted of a nucleic acid core (made of DNA or RNA) and a nucleic acid coat, or capsid, made of protein. Ozone, however, aside from its well recognized action upon unsaturated lipids, can also interact with certain proteins and their constituents, namely amino acids. Indeed, when ozone comes in contact with capsid proteins, protein hydroxides and protein hydroperoxides are formed.

Viruses have no protection against oxidative stress. Normal mammalian cells, on the other hand possess complex systems of enzymes (i.e., superoxide dismutase, catalase, peroxidase) which tend to ward off the nefarious effects of free radical species and oxidative challenge. It may thus be possible to treat infected tissues with ozone, respecting the homeostasis derived from their natural defenses, while neutralizing offending and attacking pathogen devoid of similar defenses.

The enveloped viruses are usually more sensitive to physico-chemical challenges than are naked virions. Although ozone's effects upon unsaturated lipids is one of its best documented biochemical action, ozone is known to interact with proteins, carbohydrates, and nucleic acids. This becomes especially relevant when ozone inactivation of non-enveloped virions is considered.

Fungi families inhibited and destroyed by exposure to ozone include Candida, Aspergilus, Histoplasma, Actinomycoses, and Cryptococcus. The cell walls of fungi are multilayered and are composed of approximately 80% carbohydrates and 10% of proteins and glycoproteins. The presence of many disulfide bonds has been noted, making this a possible site for oxidative inactivation by ozone.

In all likelihood, however, ozone has the capacity to diffuse through the fungal wall into the organismic cytoplasm, thus disrupting cellular organelles.

Protozoan organisms disrupted by ozone include Giardia, Cryptosporidium, and free-living amoebas, namely Acanthamoeba, Hartmonella, and Negleria. The exact mechanism through which ozone exerts anti-protozoal action has yet to be elucidated.

The positive effects of oxygenation of many dermatological conditions has long been established, and forms the basis for the use of hyperbaric oxygen treatment. Oxygen has the capacity to diffuse into the tissues, inhibit the growth of anaerobic bacteria, and raise the local oxygen content of treated tissues, thus alleviating their oxygen deprivation.

Ozone, however, as an added ingredient, has properties which clearly transcend oxygen administration alone. The two properties invoked are (I) A much broader range of pathogen killing action, and (2) A vasodilatation of arterioles, stimulating greater blood flow to tissues, with all its attendant benefits, including the greater availabilities of nutrients and of the component of vital immunological adaptations and defenses.

In view of the above-mentioned principles of external ozone/oxygen applications, we may list the following common conditions to be beneficially influenced by this unique drug therapy, utilized either in conjunction with other modalities, or used alone:

This category of wound has, by definition, not yet reached the status of chronicity due to a combination of circulatory compromise and infective onslaughts. In fact, this category of wound may simply be post-surgical, and only potentially prone to infection.

The use of topical ozone therapy in these cases may be solely preventive, and aimed at improving circulation on one hand, and inhibiting the proliferation of potentially infective organisms on the other.

Wounds which heal in an indolent manner are frustratingly difficult to master. Some of these wounds, are apt to regress, thus encouraging therapeutic strategies to become more aggressive, even experimental, but not necessarily effective.

Generally speaking, poorly healing wounds owe their definition by the chronicity of their healing, which is most commonly caused by the types and mixed variety of offending organisms they harbor.

Living organisms are constantly in contact with pathogens which, under the proper conditions, are able to parasitically proliferate to create pathological conditions. Many different types of pathogens may be involved, spanning a large spectrum of infective diversity:

Anaerobic bacteria--bacteria that do not need oxygen for their proliferation (i.e. Bacteroides, Clostridium, Streptococci), maybe noxiously active at deeper levels of the dermis, insulated to the healing influence of oxygen. Anaerobic bacteria are responsible for many devastating infections, which are generically subsumed under the appellation of gangrene. Aerobic bacteria, on the other hand, are closely identified with superficial epidermal layers; yet, when the latter are broken down, they may become influential in infective processes (i.e., Staphylococcus epidermis, Corynebacteria, Propionobacteria).

This common condition arises when a patient stays in bed, or in a wheelchair, in one position for a prolonged period of time. The pressure exerted upon the skin contact points compresses the dermal arterioles preventing proper perfusion of tissues. This leads to the oxygen starvation of tissues, impaired skin resilience, then to eventual breakdown of the skin itself. An ulcer develops, which may become quite large and usually infected with a spectrum of pathogenic organisms. At times the breakdown is so severe and the denudation of skin tissues so complete that the bottom of the ulcer reaches the bone and osteomyelitis begins.

The treatment of decubitus ulcers requires a multidisciplinary approach, including surgical, topical, and mechanico-physiological interventions. Topical antibiotics often fail to penetrate the wound and not infrequently cause secondary dermatitis in their own right.

Aside from the benefits of topical ozone therapy enunciated in this text, it should be mentioned that an added therapeutic feature of ozone, especially as it relates to the treatment of deep ulcers, is its capability to penetrate to deeper tissue level, thereby affecting pathogens which would normally be protected by tissue overlay.

This extremely common class of disorders have one common denominator, namely impaired circulation to tissues via compromise of vascular patency and integrity. A prototypic disease showing this phenomenon is diabetes. Diabetes is a complex disease which manifests both vascular disturbances to many organ systems (i.e. retina, kidney, peripheral nerves), and, in addition, disturbances to carbohydrate metabolism.

In cases where diabetes affects the peripheral circulation, tissues such as the epidermis and dermis become vascularly compromised, and thus are more prone to injuries and recalcitrant infections.

Diabetic ulcers frequently develop following simple abrasions, contusions, and lacerations. These ulcers, not unlike decubitus ulcers, are notoriously difficult to treat, and are apt to be chronically treated with topical creams and ointments, which can only address the viability of a minor proportion of putative infectious organisms. These organisms may easily develop resistance to these therapeutic agents. Concurrently, pathogens resistant to these therapies continue to proliferate and to aggravate the condition.

Ozone topical therapy, applied serially, offers the opportunity to inactivate most, if not all, offending pathogens, thus stopping the vicious cycle of infection, thus leading to ulcer healing and cicatrization. In addition, circulatory stimulation, brings essential nutritional and immunological aids to healing.

Arteriosclerosis is a condition marked by the thickening and hardening of all arterial conduits in the body. The normal pliability and patency of blood vessels is compromised, leading to disturbed circulation to many organ systems. In the case of impaired peripheral circulation (Arteriosclerosis obliterans), skin disorders may develop which include trophic changes (dry hair, shiny skin), apt to injury and eventual ulcer formation. As in the case with diabetic ulcers, these circumstances often invite multi-pathogenic infections.

The lymphatic system is essential for proper fluid equilibration within the body, and most importantly for adequate defense against infections.

Lymphedema is a condition caused by blockage to lymphatic drainage. It may be secondary to trauma, surgical procedures, and infections (i.e. streptococcal cellulitis, filiriasis, lymphogranuloma venereum).

Increasingly common is lymphedema resulting from surgical removal of lymph nodes following surgery for breast cancer. The affected arm in these patients is likely to be chronically swollen, and exercises are often prescribed to develop collateral circulation. Most importantly, however, is the occurrence of infections following even minor injuries to the arm. Injuries are then much more apt to become infected due to the absence of lymphatic system defenses. In these cases intensive topical wound care is resorted to and systemic antibiotic treatment is often prescribed.

Topical ozone treatment applied as soon as injury is noted in the affected hand or arm may prevent secondary infection, lymphedema, and the use of topical and/or systemic antibiotics.

Fungi are present on human skin in a quasi symbiotic relationship. Candida, Aspergillus, Histoplasma, are often found on intact skin, without causing clinical problems.

However, under certain conditions, the normal balance of the dermis is disturbed, allowing superficial fungi to proliferate. Tinea capitis is manifested by pustular eruptions of the scalp, with scaling and bald patches. Tinea cruris is a fungal pruritic dermatitis in the inguinal region.

Serial topical ozone applications have shown marked success in eradicating the most chronic and stubborn fungal skin conditions.

Thermal burns are divided into first, second, and third degrees, depending upon the depth of tissue damage. First degree burns are superficial, and include erythema, swelling, and pain. In second degree burns, the epidermis and some portion of the underlying dermis are damaged, leading to blister and ulcer formation. Healing occurs in one to three weeks, usually leading to little or no scar formation.

In third degree burns, muscle tissue and bone may be involved, and secondary infection is very common.

It is in cases marked by significant tissue injury, and especially in cases involving infections, that topical ozone therapy finds the most usefulness. In the case of burns, the range of pathogenic organisms may be extremely wide (see the section on poorly healing wounds), and thus may be ideally suited for ozone therapy.

Ozone is actively virucidal to a staggering number of viral families. Most clearly documented are ozone's neutralizing effects upon lipidenveloped virions. These include diverse viral groups as the Hepadnaviridae (Hepatitis B and C), the Retroviridae (HIV-I and HIV-II), the Herpesviridae (Herpes simplex I and II, Cytomegalovirus, Epstein-Barr), Filoviridae (Ebola virus and Marburg virus), Orthomyxoviridae (Influenza A and B ), the Paramyxoviridae (Measles, Mumps, Parainfluenza, Respiratory syncytial virus), the Coranoviridae, the Togaviridae (Rubella, Eastern and Western equine encephalitis), and the Rhabdoviridae (Rabies).

Although lipid-enveloped viruses appear to be most susceptible to ozone inactivation due to their dependency on their outer lipid sheath, non-enveloped viruses are also negatively subject to ozone through its ability to interact with proteins, amino acids, carbohydrates and glycoproteins.

Herpes simplex viruses are extremely widespread in the human population. Two distinct types of viruses are known, namely Herpes simplex type I and II. Type I is transmitted via contact through mucosa or broken skin (often through saliva), while type II is more specifically sexually propagated.

In herpetic lesions, fluid accumulates between the dermis and epidermis, producing vesicles which rupture, thus releasing more virions. They then become easily infected by secondary organisms.

Herpes lesions have been extensively studied with reference to topical ozone administration. Ozone in these cases (I) Directly inactivated herpes viruses which are lipid-enveloped (2) Act as a pan-bactericidal agent in cases involving secondary infections, and (3) Promotes healing of tissues through circulatory enhancement. It is also postulated that ozone may have beneficial effects upon the peripheral neurons which harbor these viruses.

Afflictions implicating nails which are therapeutically assisted by topical ozone treatment include the following:

1. Candida albicans. Nails in this condition are painful, with swelling of the nail fold, and often, thickening and transverse grooving of the nail architecture. Loss of the nail itself is not infrequent. Another frequent condition is Tinea Unguium, marked by thickened, hypertrophic, and dystrophic toenails. There are currently no antifungal agents of proven efficacy for this condition.
2. Tinea Pedis (Athlete's Foot). This very common disorder is caused by infection with species of Trichophyton, and with Epidermophyton floccosum. Chronic infection involving the webbing of the toes may evolve to secondary bacterial involvement. Lymphangitis and lymphadenitis may present themselves, as well as infection of the nails themselves (Tinea unguium, Onychomycosis). Nails may become thickened, yellow, and brittle. The patient may then develop allergic hypersensitivity to these organisms which may manifest in other parts of their bodies.

Topical ozone therapy offers unique treatment opportunities to these recalcitrant infections. Ozone penetrates the affected areas, including the nails proper, and with repeated administration, is capable of inactivating all species of fungi mentioned above.

Healing occurs slowly yet consistently, and skin integrity along with nail anatomy, gradually regain their normal configuration.

This condition occurs during times when the body is exposed to ionizing radiation. This may occur during an accident, or within the course of radiation therapy. Radiation energy is imparted to individual cells, leading to alteration in cellular DNA, thus favoring cellular injury and/or death.

Clinical findings are commensurate with the type, amount, and duration of radiation exposure. Several clinical syndromes have been delineated, including Radiation Erythema, Acute Radiodermatitis, and Chronic Radiodermatitis.

While DNA damage cannot be easily repaired (except perhaps partially through nutritional avenues such as vitamin E), secondary infections made more likely by decreased tissue resistance, may be countered by topical ozone therapy. This avoids the systemic absorption of topical creams and ointments, and ensures pan-pathogen protection.

Factors contributing to skin injuries due to cold derive from vasoconstriction and the formation of ice crystals within tissues. As frostbite progresses, loss of sensation occurs, and tissues become increasingly hard to the touch. Depending upon length of exposure and processes related to rewarming, dry gangrene may develop. Dry gangrene may evolve to wet gangrene if infection occurs.

Topical ozone therapy has proven to be effective in decelerating or halting the pathogenesis of frostbite through (I) The immediate oxygenation of tissues, (2) Increasing blood flow through a direct vasodilatory effect upon the dermal arterioles, and (3) The prevention of secondary infection.

Topical ozone therapy for the disorders mentioned above requires sophisticated medical diagnosis of the underlying conditions, and an appropriately tailored treatment plan, which may include any one of several therapeutic modalities utilized concomitantly, including ozone, or may call for the utilization of ozone as the sole therapeutic intervention.

1. The ease of administration of this therapy, taking into consideration the strict parameters of the technology-to-drug symbiosis.
2. Ozone is an effective antagonist to the viability of an enormous range of pathogenic organisms. In this regard, ozone cannot be equaled. It is effective in inactivating anaerobic and aerobic bacterial organisms, a wide spectrum of viral particles--lipid as well as non-lipid enveloped--and a substantial spectrum of fungal and protozoal pathogens.

To replicate this therapeutic action, the medical conditions in question would have to be treated with a conglomeration of antibiotic agents, systemically and/or topically applied. This would present, in the context of contemporary medical practice, massive clinical difficulties.

3. Ozone therapy, appropriately applied in a timely fashion, may obviate the need for systemic anti-pathogen therapy, thus saving the patient from all the side effects and organ stresses this option could entail.
4. Ozone exerts its anti pan-pathonegic actions through entirely different mechanisms than conventional antibiotic agents. The latter must be constantly upgraded to surmount pathogen resistance and mutational defenses. Ozone, on the other hand, presents direct oxidative challenge which cannot, by all available pathogen defenses cannot be circumvented.

Topical ozone therapy has shown effectiveness in an impressive array of medical conditions. In this article, the following are cited: Infected wounds; poorly healing wounds; decubitus ulcers; circulatory disorders; lymphatic diseases; fungal skin infections; burns; cutaneous viral afflictions; nail afflictions; radiodermatitis; and frostbite.

Ozone presents many features that are common to many drugs, namely a therapeutic window demarcated by sub-optimal dosage on one hand, and toxic higher dose levels on the other. For this reason ozone dosage must be carefully calibrated and delivered, a feasibility which has only currently been achieved through advances in contemporary technology.

Ozone is a pan-bactericidal, pan-virucidal, anti-fungal and antiprotozoan therapeutic agent which, utilized under treatment protocols which continue to need proper delineation through research, promises to become a potent adjunct to current medical treatment. It is also likely to show promise as a drug used as a sole therapeutic agent in our global growing need to bolster our antipathogen armamentarium.

• Buckley R D, Hackney J D, Clarck K, Posin 1975 Ozone and human blood. Archives of Environmental Health 30:40-43
• Cann A J 1997 Principles of Molecular Virology. Academic Press, San Diego
• Champion R H, Burton J L, Ebling F J,1992 Textbook of Dermatology, Blackwell Scientific Publications, Oxford
• De Groot A C, Weyland W J, Nater J P,1994 Unwanted Effects of Cosmetics and Drugs Used in Dermatology, Elsevier, Amsterdam
• Habif T P Clinical Dermatology 1966, Mosby, St Louis
• Dyas A, Boughton B, Das B 1983 Ozone killing action against bacterial and fungal species. Journal of Clinical Pathology 36(10):1102-1104
• Epstein E 1994 Common Skin Disorders, Saunders, Philadelphia
• Evans A S, Kaslow R A (Eds) 1997 Viral infections of humans. Epidemiology and control. Plenum, New York
• Farooq S, Akhlaque S 1983 Comparative response of mixed cultures and virus to ozonation. Water Research 17:809
• Harakeh M, Butler M J 1985 Factors influencing the ozone inactivation of enteric viruses in effluent. Ozone: Science and Engineering 6:235-243
• Langlais B, Perrine D 1986 Action of ozone on trophozoites and free amoeba cysts, whether pathogenic or not. Ozone: Science and Engineering 8:187-198
• Leland D S 1996 Clinical virology. Saunders, Philadelphia
• Marhell E K, Voge M, John D T 1986 Medical Parasitology. Saunders, Philadelphia
• Menzel D 1984 Ozone: An overview of its toxicity in man and animals. Toxicology and Environmental Health 13:183-204
• Murray P R (Ed) 1995 Manual of Clinical Microbiology. ASM Press, Washington D.C.
• Razumovskii S D, Zaikov G E, 1984 Ozone and its reactions with organic compounds. Elsevier, Amsterdam
• Roy D, Wong P K, Engelbrecht R S, Chian E S 1981 Mechanisms of enteroviral inactivation by ozone. Applied Environmental Microbiology 41:728-723
• Ryan K J (Ed) 1994 Medical Microbiology. Appleton & Lange, Norwalk, Connecticut
• Shelley W B, Shelley E D, 1992 Advanced Dermatological Diagnosis, Saunders, Philadelphia
• Sobsey M D 1989 Inactivation of health-related microorganisms in water by disinfection processes. Water Science Technology 21(3)179-195
• Sunnen G V 1988 Ozone in medicine: Overview and future directions. Journal of Advancement in Medicine 1(3):159-174
• Vaughn J M, Chen Y, Linburg K, Morales D 1987 Inactivation of human and simian rotaviruses by ozone. Applied Environmental Microbiology 48:2218-2221
• Viebahn R 1994 The use of ozone in medicine. Haug, Heildelberg
• Werkmeister H 1985 Subatmospheric 02/03 treatment of therapy-resistant wounds and ulcerations. OzoNachrichten 4:53-59

This is an addendum to a February/March 1994
article on Ozone in Medicine.

The inactivation of viral particles by ozone may take place by a variety of mechanisms which range from direct physico-chemical effects to more indirect immunological pathways. Virions coated by a lipid glycoprotein envelope such as rectoviruses, hepatitis B and C, Herpes 1 and 2, and Epstein-Barr among others, are vulnerable to the influence of ozone by its intense oxidizing properties,

In retroviruses for example, which possess a glycolipid encapsulation, ozone confronts the double bond sites in its matrix thus destroying its architecture. Without its envelope, the virus perishes.(60) In virions which lack a lipid envelope but whose nucleic acids are surrounded by a protein capsid such as those of the minovirus family, ozone may diffuse through the protein coating and deform or cleave the genome core. Viruses, unlike cells, lack enzymes designed to repair injured DNA or RNA, and are incapacitated by this process.

In major autohemotherapy, relatively large amounts of blood are treated with ozone, then reinfused into the patient. The dosage and concentration of ozone administered is carefully calibrated so that maximal antiviral action is mobilized while at the same time sparing the integrity and viability of the cellular elements. Viruses are small, denuded of complex defenses,(63) and vulnerable to the oxidative challenge of ozone. Cells, in contrast, are large and incorporate a multiplicity of homeostatic mechanisms. There are developing technologies which are designed to treat whole blood in a manner similar to the dialysis process.

In the case of retroviral infection, it is extracellular viral particles which are presumably most affected by ozone oxidation. Intracellular virions or provirions on the other hand, were thought to be relatively spared of destruction by ozone by the barrier of cellular membranes, the buffer of cytoplasmic constituents, and by the refuge provided by their incorporation into the genome of the host cell. However, it has been demonstrated that ozone possesses the capacity to inactivate intracellular virions as well.(64)

Major autohemotherapy, repeatedly administered, could thus exert a culling action on circulating virions, especially during phases of the viral life cycle associated with viral seeding in the general circulation, the so-called viremic episodes. Studies attempting to measure the efficacy of this treatment modality should take into account the patient's clinical status as it relates to the cycle phase of viral activity at the time of the therapeutic intervention.

Immunological mechanisms may be invoked through several pathways. In minor autohemotherapy, a small amount of blood is ozone-treated in such a manner as to fragment most virions without regard to preserving cellular elements. This treated blood, injected intramuscularly, carries fragments of viral envelope and nucleic acids which find their way into the general circulation and to the immune network. The latter, if still relatively operational, begins to manufacture appropriate antibodies which in turn, serve to counter the evolution of the infection.

The interesting feature of this technique is that antibodies thus manufactured are individualized to the particular patient receiving the treatment, since they are derived from their own viral stock, In view of the high mutability of retroviruses, each patient carries a unique viral strain. Minor autohemotherapy can thus be conceptualized as a method of autovaccination providing a high degree of antibody specificity.

A non-invasive and increasingly popular method of oxygen/ozone administration, the so-called "Sauna bag" method, does not involve heat, but consists of enclosing the patient up to the shoulders with a comfortable ozone-resistant plastic cover. An oxygen/ozone mixture is introduced in the bag, and' the patient allows it to interface with the entire skin surface for a few minutes. Surprisingly, the mixture is able to penetrate far enough into the capillary networks to raise blood oxygen pressure. Presumably then, ozone is able to exert its biochemical influence. The added advantage in this technique is that superficial skin conditions amenable to antiseptic influence are addressed. As with all ozone therapies, gas mixture concentration and duration of exposure need to be clinically adjusted and monitored.

In recent years, it has been discovered that nitric oxide, a gas under atmospheric conditions traditionally associated with toxicity, actually exerts essential biological functions. It has a free radical structure, is short-lived, and is an eager electron contributor. Aside from its activity as a neurotransmitter and as an antihypertensive agent, nitric oxide appears to be an essential component of the mechanisms by which macrophages become activated to destroy tumor cells, bacteria and viruses. Macrophages, scavenger components of the immune network, become activated by creating minuscule amounts of nitric oxide using arginine as a substrate and the enzyme nitric oxide synthase. Without nitrid oxide, macrophages remain idle. It has also been shown that nitric oxide is directly toxic to tumor cells.(62)

It may be theorized that ozone with the mobilization of its own free radical structure could facilitate the elaboration of nitric oxide in macrophages, thus promoting their scavenging mission.

In January 1994, the first Phase 1 (human) clinical trial of ozone therapy will be conducted at 5 major University centers in Italy. It will involve 300 volunteers, will be conducted according to FDA-approved protocols, and will test the effectiveness of major auto-hemotherapy in AIDS and Hepatitis B. The scientific community is eagerly awaiting the data generated by this breakthrough study.

Ozone's interactions with biological systems and its activities in pathogen inactivation are varied, complex, and to a large extent still largely unknown. The recent discoveries that nitne oxide and carBon monoxide(61)(62) assume crucial functions in regulating metabolic and physiological health, may give new reasearch impetus to the investigation of the therapeutic properties of ozone.

Gérard V. Sunnen, M.D, 200 East 33rd St. #26J New York, NY 10016 212-679-0679

Editor's Comment: The original article "Ozone in Medicine" (Feb/Mar 1994 TLfD) was previously published in the Journal o Advancement in Medicine (1988).

References

60. Wells K, Latino J, Gavalchin J, Poiesz B: Inactivation of Human lmmunodeficiency Virus Type 1 by Ozone in vitro. Blood. Oct. 1, 1991;78(7):1882-1890.

61. Verma A, Hirsch D, Glatt C, Ronnett G, Snyder S: Carbon Monoxide: A Putative Neural Messenger. Science. 15 Jan 1993:259(5093):381-384.

62. Snyder S. Bredt D: Biological Role of Nitric Oxide. Scientific American. May 1992;266(5):68-77.

63. Evans E. ed: Viral Infections of Humans. 1991. 3rd Edition. Plenum Medical book Company. New York and London.

64.Baggs A: Are Worry-free Transfusions Just a Whiff of Ozone Away? Can Med Assoc J. 1993:148(7):1156-1160

65.Carpendale M, Griffiss J: Is There a Role for Medical Ozone in the Treatment of HIV and Associated Infections? Proceedings, Eleventh Ozone World Congress, San Francisco, 1993.

Comentaries on Prion Diseases and Ozone

by Gérard V. Sunnen, M.D.

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A number of diseases afflicting humans, animals and plants are, in contemporary times, stimulating great interest. This is due to the fact that these conditions do not follow traditional trajectories of infectivity, and that the responsible infective agents have poorly understood structural configurations and/or mechanisms of action.

Transmissible Spongiform Encephalopathies (TSEs) occur in humans and animals, and are characterized by progressive pathological effects upon the nervous system, invariably resulting in death. As their names suggest, they produce, usually over long periods of time, neuronal loss and gliosis, leading to microlacunae in brain tissue, which are apt to fill, by unknown mechanisms, with amyloid-like deposits.

Human TSEs include Creuzfeldt-Jacob disease (CJD), Familial fatal insomnia (FFI), Kuru, and Gerstmann-Straussler-Scheinker disease (GSS).

Animal TSEs include Scrapie in sheep, Feline spongiform encephalopathy (FSE), Chronic wasting disease in deer and ungulates, and of most concern in current times, Bovine spongiform encephalopathy (BSE). This concern is derived from evidence of transmission of TSEs from animals to humans.

These diseases were suspected of having viral origins, until the 1960's when T Alper suggested that the scrapie agent might have the capacity of replicating without the presence of nucleic acid. In 1982, S Prusiner coined the word prion, to stand for a new class of infective agents with highly novel characteristics.

Prions are not conventional viruses. Nucleic acid is apparently not necessary for their infectivity. In addition, they appear to interact with host genes, cell proteins, and cellular strucures in very complex and yet unchartered fashion.

Properties which further differentiate prions from viruses include their resistance to radiation, to heat inactivation, to DNAse and RNAse treatment, and to such protein-denaturing chemicals as phenols. These characteristics point to a protein structure in prions. Some prions have been shown to consist of glycoproteins with amino acid sequences approximating 250 units. There is no evidence at this time that prions contain lipid components.

It appears, interestingly, that, aside from the precise sequencing of amino acids within their structure, the spatial configuration of prions may play an important role in their pathogenecity, especially as it relates to the disruption of neuronal membranes. There is evidence that this is true in the case of scrapie prions which contain two highly hydrophobic regions capable of spanning such membranes. Presumably, an alteration in the molecular architecture of the prion could impair this capacity for membrane attachement, and thus compromise its destructive potential.

Prions, by all current evidence, are very precisely structured molecules, whose specific stereotaxis needs to be quintessentially intact for the expression of their infectivity.

Ozone has not yet been studied for the inactivation of prions or viroids (such as HDV). While the agents cited above such as heat, radiation, enzymes, and cleaving chemicals, have proven to be unsuccessful in inactivating prions, ozone has properties which are sufficiently distinct from these agents to embody a unique potential in offering novel mechanisms of prion destruction. Specifically, ozone has the proven ability to offer intense oxidizing action, which may provide a means of altering the prion's precisely tuned chemical and spatial design through:

1. A demonstrated capacity to react with C-C bonds of organic origin, resulting in:
2. The selective breakage of multiple bond linkages, thus permanently altering the crucially needed proper sequencing of amino acid units, thus inducing a probable alteration in their proper attachment and alignment to each other, and to associated components such as carbohydrate-and possibly lipids-and,
3. The metamorphosis of the spatial configuration of the prion in its globality, which, experimental data shows, could have major implications in the mechanisms of its reproductive strategy, It is theoretically plausible that in compact molecular structures such as prions, whose amino acid sequences have been so precisely elaborated over long periods of time, that even miniscule alterations in their composition and/or configuration could insure their deactivation.
4. Ozone's ability, when applied to fluids, to react with components of these fluids in the entirety of their volume and space, almost instantaneously. Ozone, as a gas, follows the laws of gas to liquid dynamics, as is thus distinctly and intrinsically different from all other inactivating agents.

From: Cann A J 1997 Principles of Molecular Virology. Academic Press, San Diego

The illustration shows two different spatial configurations of the same prion structure. The one on the right is infectious and pathogenic, while the other is not. The factors which promote prions to metamorphose their spatial architecture are unknown.

In conclusion, ozone offers unique capabilities to potentially destroy prions. Ozone, while altering these small infective protein molecules, is theoretically capable of exerting its contrapathogenic role, while leaving much larger and versatile protein and lipid molecules found in mammalian serum functionally intact.

BIBLIOGRAPHY

• Cann A J 1997 Principles of Molecular Virology. Academic Press, San Diego
• Diener T 0 1987. The Viroids. Plenum Press, New York
• Eigen M 1996. Prionics or the kinetic basis of prion diseases. Biophysiological Chemistry Dec 10;63(1):1-18
• Fleminger S et al Prion diseases 1997. British Journal of Psychiatry 170:103-105
• Harrison P M 1997. The prion folding problem. Current Opinion in Structural Biology. Feb 7(1):53-59
• Horwick A L et al 1997. Deadly conformations-protein misfolding in prion diseases. Cell. May 16;89(4):449-510
• McCardle L 1997. Human prion diseases. British Journal of Biomedical Science 54(1):2-4
• Prusiner S B 1995. The prion diseases. Scientific American 272:48-57
• Razumovskii S D, Zaikov G E 1984. Ozone and its reactions with organic compounds. Elsevier, Amsterdam
• Taylor D 1996. Inactivation of the causal agents of Creutzfeldt-Jacob disease and other prion diseases. Brain pathology 6(2):197-198
• Will R G et al 1996. A new variant of Creutzfeldt-Jakob disease in the UK. Lancet 347:921-925

February 2001

Abstract

Summary

BIBLIOGRAPHY

• Bartenschlager R. Candidate targets for hepatitis C virus-specific antiviral therapy. Intervirology 1997; 40:378-393
• Bocci V, Luzzi E, Corradeschi F, Paulesu, et al. Studies on the biological effects of ozone: 5. Evaluation of immunological parameters and tolerability in normal volunteers receiving ambulatory autohaemotherapy. Biotherapy 1994; 7:83-90
• Bocci V. Ozonation of blood for the therapy of viral diseases and immunodeficiencies. A hypothesis. Medical Hypotheses 1992 Sept; 39(1):30-34
• Bolton DC, Zee YC, Osebold JW. The biological effects of ozone on representative members of five groups of animal viruses. Environmental Research 1982; 27:476-48
• Buckley RD, Hackney JD, Clarck K, Posin C. Ozone and human blood. Archives of Environmental Health 1975; 30:40-43
• Cardile V, et al. Effects of ozone on some biological activities of cells in vitro. Cell Biology and Toxicology 1995 Feb; 11(1):11-21
• Carpendale MT, Freeberg JK. Ozone inactivates HIV at noncytotoxic concentrations. Antiviral Research 1991; 16:281-292
• Dailey JF. Blood. Medical Consulting Group, Arlington MA, 1998
• Di Bisceglie AM, Bacon BR. The unmet challenge of hepatitis C. Scientific American 1999; 281:58-63
• Dienstag JL. Sexual and perinatal transmission of hepatitis C. Hepatology 1997; 26: 66S-70S
• Dieperink E, Willenbring M, Ho SB. Neuropsychiatric symptoms associated with hepatitis and interferon alpha: A review. Am J Psychiatry 2000 June; 157(6):867-876
• Evans AS, Kaslow RA (Eds). Viral Infections in Humans: Epidemiology and Control, Fourth Edition, Plenum, New York, 1997
• Gonzalez-Peralta RP, Qian K, She JY, et al. Clinical implications of viral quasispecies heterogeneity in chronic hepatitis C. J Med Virology 1996; 49:242-24
• Harrison TJ, Zuckerman AJ (Eds). The Molecular Medicine of Viral Hepatitis. Molecular Medical Science Series. John Wiley & Sons, New York, 1997
• Konrad H. Ozone therapy for viral diseases. In: Proceedings 10th Ozone World Congress 19-21 Mar 1991, Monaco. Zurich: International Ozone Association 1991:75-83
• Liang TJ, Hoofnagle JH, (Eds). Hepatitis C. Academic Press, San Diego, 2000
• Maggi F, Fornai C, Morrica A, et al. Divergent evolution of hepatitis C virus in liver and peripheral blood mononuclear cells of infected patients. J Med Virology 1999; 57:57-63
• Major ME, Feinstone SM. The molecular virology of hepatitis C. Hepatology 1997; 25: 1527-1538
• Maertens G, Stuyver L. Genotypes and genetic variation of hepatitis C virus. In: The Molecular Medicine of Viral Hepatitis. John Wiley $ Sons Ltd., London, 1997: 225-227
• Monjardino J. Molecular Biology of Hepatitis Viruses, Imperial College Press, London, 1998
• Par A, et al. Hepatitis C virus infection: pathogenesis, diagnosis and treatment. Scandinavian Journal of Gastroenterology. 1998 Suppl; 228: 107-114
• Paulesu L, Luzzi L, Bocci V. Studies on the biological effects of ozone: Induction of tumor necrosis factor (TNF-alpha) on human leucocytes. Lymphokine Cytokine Research 1991; 5:409-412
• Pawlotsky J. Hepatitis C virus resistance to antiviral therapy. Hepatology Nov. 5, 2000; 32: 889-89
• Roy D, Wong PK, Engelbrecht RS, Chian ES. Mechanism of enteroviral inactivation by ozone. Applied Environmental Microbiology 1981; 41: 728-733
• Sarara AI. Chronic hepatitis C. South Med J. 1997; 90: 872-877
• Seeff LB. Natural history of hepatitis C. Hepatology 1997; 26: 21S-28S
• Sunnen GV. Ozone in Medicine. Journal of Advancement in Medicine. 1988 Fall; 1(3): 159-174
• Trivedi M. Newly diagnosed hepatitis C: Lack of symptoms doesn't mean lack of progression. Postgraduate Medicine 1997; 102: 95-98
• Valentine GS, Foote CS, Greenberg A, Liebman JF (Eds). Active Oxygen in Biochemistry. Blackie Academic and Professional, London, 1995
• Vaughn JM, Chen Y, Linburg K, Morales D. Inactivation of human and simian rotaviruses by ozone. Applied Environmental Microbiology 1987; 48:2218-2221
• Viebahn R. The Use of Ozone in Medicine. Haug, Heildelberg, 1994
• Wells KH, Latino J, Gavalchin J, Poiesz BJ. Inactivation of human immunodeficiency virus Type 1 by ozone in vitro. Blood 1991 Oct; 78(7):1882-1890
• Yu BP. Cellular defenses against damage from reactive oxygen species. Physiological Reviews 1994 Jan; 74(1):139-162

First published September 22, 1997

The Nature of Viruses

Viruses are intracellular parasites. They require a living host cell in order to replicate and to infect new hosts. Viruses have been enormously successful in parasitizing most known forms of living organisms in both the animal and plant world.

Components of Viruses

1. Nucleic acid core. At the core of viruses is genetic material which encodes the transcription of all viral components, mostly proteins (such as enzymes). This genetic material is either RNA or DNA, never both. The nucleic acid may be single or double stranded. Viral nucleic acid has been described as the software program for making copies of the virus.
2. The nucleic acid coat or capsid. Surrounding the core is a protective coating made of protein, called the capsid. The capsid is rigid and determines the shape of the virus. It is made of repeating protein units called capsomeres.

The architecture of capsomere assembly is quite fascinating. A very common crystal-like configuration is the 20-sided isocahedral construction, with each capsomere forming an equilateral triangle.

Another main pattern is helical or rod shape. The nucleic material and its capsid curls unto itself like a snake, forming a spherical structure.

A few viruses have capsids that are neither icosahedral nor helical; these configurations are termed complex, and may be spiral, brick shaped, or may show other non-standard appearances.

The capsid and its nucleic acid core together form the nucleocapsid.

3. The nucleocapsid is sometimes surrounded a membrane called an envelope. Enveloped viruses are usually spherical because the envelope, unlike the capsid, is loose-fitting. Envelopes are lipid bilayers that contain proteins. Some proteins incorporate carbohydrates and are thus called glycoproteins. Glycoproteins usually protrude out of the envelope as spikes called peplomers. The function of peplomers is to form points of attachments to host cell receptors for entry into cells.

FIGURE 1-1 SIZE OF MICROSCOPIC ENTITIES AND MICROSCOPE RESOLUTION. Viruses are smaller than the smallest bacteria and larger than macromolecules. They can be seen with the electron microscope. (Illustration demonstrates relative sizes and is not drawn to scale.) From: Leland DS 1996 Clinical Virology, Saunders, New York

FIGURE 1-2 ICOSAHEDRAL CAPSID CONFIGURATION IN NAKED AND ENVELOPED VIRUSES. Naked icosahedral viruses appear cubic or crystalline (A, B, and C). Enveloped icosahedral viruses appear nearly spherical (D, E, F). (Fig. 1-2C from Ryan KJ (Ed), Sherris Medical Microbiology: An Introduction to Infectious Diseases, 3rd ed. Norwalk, CT: Appleton & Lange, 1994. Fig. 1-2F From: Murray PR, Kobayashi GS, Pfaller MA, Rosenthal KS. Medical Microbiology, 2nd ed. St. Louis: Mosby Year Book, 1994, p 573

FIGURE 1-3 HELICAL CAPSID CONFIGURATION IN NAKED AND ENVELOPED VIRUSES. Naked helical viruses are cylindrical or rod shaped (A, B, and C). Enveloped helical viruses appear nearly spherical because the nucleocapsid (capsid and nucleic acid) may curl inside the envelope (D, E, and F). Fig. 1-3C from: Tortora et al. Microbiology, 4th ed., Benjamin Cummings, Redwood City CA, 1992, p 336. Fig. 1-31F From: Ryan KJ (Ed), Sherris Medical Microbiology: An Introduction to Infectious Diseases, 3rd ed. , Appleton & Lange, Norwalk CT, 1994.

The envelope is sometimes connected to the nucleocapsid by a matrix made of proteins (matrix proteins).

4. Viruses that do not have an envelope are called "naked viruses".
5. The term virion describes the mature viral particle capable of infecting other cells. In non-enveloped viruses, the virion consists of the nucleocapsid alone. In enveloped viruses, the nucleocapsid and the envelope make up the virion.

Enveloped viruses are usually comfortable in bodily fluids and are transmitted by such routes as blood transfusion, or mucosa to fluid contact, as in sexual contact.

Naked viruses, on the other hand, are usually transmitted via the intestinal route.

Major Enveloped Viruses Include the Following Families:

Major Non-Enveloped (Naked) Viruses:

The Viral Life Cycle

Viruses have complex life cycles which demonstrate their extraordinary symbiosis with their hosts. They lack the tools for self sufficient growth and thus depend upon more advanced life systems for their existence.

Viral replication, in broad strokes, starts with viral attachment to host cells. Enveloped viruses use glycoprotein molecules on surface peplomers for binding to the cell membrane. Viral and cell membranes fuse and the viral nucleocapsid penetrates into the cell's cytoplasm. Naked viruses may be engulfed by the cell membrane in a process called endocytosis. Once inside the cell, the capsid is reconfigured to prepare it for replication.

In the replication phase, viral nucleic acid has the capacity to direct the host cell to manufacture all components of the mature virion. These are then assembled and mass produced. The virions are then released into the circulation in a process that may or may not involve cell destruction , or lysis.

A wave of viral particle carried through the blood stream to infect organs throughout the body is called a viremic episode. Many viruses travel free in the plasma. Others may attach themselves to platelets, lymphocytes or red blood cells.

Recent research has shown that the number of virions involved in infection is much more important than previously realized. In both HIV and hepatitis C, several billion new particles may be produced each day. The amount of viral concentration present at any one time is called the viral load. The immune system is placed under constant stress to deactivate these new infective particles, and to regenerate its own decimated cellular components.

Any reduction in the viral load offers an advantage to the immune system, and thus enhances the probability for easing clinical symptoms.

Commentaries on the Evolution of Viruses

Viruses are far from being static entities. As quintessential intracellular parasites they have developed, through millions of years of cohabitation with their hosts, astoundingly sophisticated structures, survival, and propagation mechanisms. They have adapted, modified their biological strategies, and incorporated genetic diversity and mutational capacity to cope with the changing ecology.

In the twentieth century, this ecology, namely the human reservoir, has changed dramatically. The eruptive world population and the mobility of the planet's inhabitants are two major factors responsible for the accelerated evolution of viruses into new frontiers of pathogenicity.

Most of the families of viruses mentioned above have produced new strains. Of great concern is the diversification of retroviruses and of Hepatitis B and C. Since 1985, for example, the following viruses are among the many that have been discovered: Human herpesvirus 6, 7, and 8; Hepatitis C; Hepatitis E; Morbillivirus, causing encephalitis; Hantaviruses causing the hantavirus pulmonary syndrome.

Principles of Viral Inactivation

The inactivation of viral particles in the test tube is possible by a number of interventions. The enveloped viruses are usually more sensitive to physico-chemical challenges than are naked virions. Chemical agents and other adverse conditions that affect the envelope invariably destroy the entire virus. These include drying, high temperature, freezing and thawing, pH below 6 or above 8, lipid solvents, hydrogen peroxide, chloroform, chemicals containing chlorine, ultraviolet irradiation, phenols, and ozone.

The test tube environment presents different parameters than, let us say, a biologically active substance e.g.., a serum, or a vaccine. In this case the virions in solution need to be destroyed while the components of the biological solution need to be preserved.

Ozone is an exemplary agent capable of deactivating a wide range of viruses while significantly sparing the biological integrity of their medium. There is, as concerns ozone, a therapeutic range of administration. Like many other drugs, ozone may be said to have a therapeutic window which defines its optimal levels of administration. Below the window, concentrations of administered ozone are poorly effective; within the window they function optimally; and above the window, deleterious effects take place.

Ozone offer distinct advantages over other antiviral agents. It is above all, a gas. When a gas is administered to a liquid, an immediate dispersion of the gas occurs, thus effecting reactions in the entire fluid volume. This may be contrasted to any fluid additive, such as hydrogen peroxide or detergents which mix with the fluid being treated much more slowly due to the physics of fluid-fluid dynamics. Furthermore, at the interface of the fluids, the concentrations of the chemicals are inordinately high and potentially toxic. As concerns the viability of cellular elements such as red and white blood cells, the oxidizing potential of ozone may be accurately calibrated so that it is fatal to virions, but innocuous to cells. Cells protect themselves from oxidative injury by means of protective enzymes (glutathione, superoxide dismutase, catalase), in contrast to virions which have no protection against oxidation.

The enveloped viruses are more fragile than the naked viruses. Their envelope, made up of lipids and glycoproteins are especially vulnerable to ozone's capacity for oxidation (oxidation is defined as the removal of electrons from compounds). Envelope lipids which are unsaturated, when exposed to ozone, become saturated, are cleaved at their double bond sites, and break apart. The envelope thus becomes torn and the viral capsid, by itself, cannot survive.

Although ozone's effects upon unsaturated lipids is one of its best documented biochemical actions, ozone is known to interact with proteins, carbohydrates, and nucleic acids. This becomes especially interesting when ozone inactivation of non-enveloped virions is considered.

The protective layer surrounding the DNA or RNA of virions (the capsid), is made up of proteins. Circulating freely in the bloodstream, the capsid's protein coat, in the case of naked viruses, is thus its first and last line of defense. Challenged by ambient ozone or its peroxides, the protein coating itself becomes denatured and incapable of sustaining its protective role (The viral nucleic acid material, by itself cannot survive). Indeed, when ozone comes in contact with capsid proteins, protein hydroxides and protein hydroperoxides are formed.

In the viral decontamination of a serum sample, it is assumed that several viral species may be present. Each virus has its own tolerance, and intolerance, for ozone challenge. There is an optimal concentration range, however, within which ozone may be administered to the serum sample, destroying its viral and bacterial occupants, and at the same time preserving the great majority of its biological activity (i.e., in the case of, vaccines, antigen purity; or in passive immunization, the sustenance of antibody titers).

The task of viral inactivation in vivo becomes more difficult. In this case we are dealing with a live patient and the awesomely complex dynamics determining the evolution of a viral infection. Some general guiding principles nevertheless stand out:

We have seen that viremic episodes represent invasions of virions into bodily fluids. In the case of acute infections such as Ebola, or more commonly in the flu syndrome, there may be one viremic episode, which in the first instance may be fatal, and in the second, may be hardly noticed by the patient. depending upon host factors.

In the case of chronic infections such as hepatitis or HIV, however, viremic episodes may occur numerous times in periods spanning several years. Viral load may be high, indicating a shift towards virus victory in relation to immune surveillance, defense and reserve, or may be low, indicating a quiescence within the viral life cycle.

Any intervention which will safely decrease the numbers of virions from the circulation will proffer an advantage to beleaguered immune function.

Ozone hemotherapy consists in the regular treatment of aliquots of blood with precise doses of ozone. The result is a culling action due to the direct action of ozone and the biologically active compounds it produces on viral particles, which translates into a progressive diminution in the viral load, and a corresponding enhancement of immune potency.

Another mechanism of viral inactivation with ozone is indirect. Subsequent to the treatment of blood with ozone, there exists in the serum a plethora of fragmented virions most of which are excreted through the kidneys. Some of these fragments, however, are processed by the immune system for the elaboration of its own defenses.

We know that each HIV-afflicted patient, for example, is infected with a unique subspecies of HIV virion. Both intact virions - and once destroyed, their fragments - have one of a kind antigenic structures (an antigen is defined as a substance capable of stimulating the production of antibodies). The immune system is thus able to manufacture organism-specific antibodies. The still poorly appreciated uniqueness of ozone therapy in this regard, is that - assuming the preservation of a minimum of immunocompetence - it provides the patient with an opportunity to make his own individualized autovaccine to the distinctive type of virus particle harbored.

Ozone: Clinical Methodology Ozone may be utilized for the therapy of a spectrum of clinical conditions. Routes of administration are varied and include external and internal (blood interfacing) methods. In the technique of ozone major autohemotherapy for viral diseases, an aliquot of blood is withdrawn from a virally-afflicted patient, anticoagulated, interfaced with an ozone/oxygen mixture, then re-infused. This process is repeated serially until viral load reduction is documented.

Ozone: A Review of Possible Mechanisms of Anti-viral Action

• Denaturation of virions through direct contact with ozone. Ozone, via this mechanism, disrupts viral envelope proteins, lipoproteins, lipids, and glycoproteins. The presence of numerous double bonds in these unsaturated molecules makes them vulnerable to the oxidizing effects of ozone which readily donates its oxygen atom and accepts electrons in these redox reactions. Double bonds are thus reconfigured, molecular architecture is disrupted and widespread breakage of the envelope ensues. Deprived of an envelope, virions cannot sustain nor replicate themselves.
• Ozone proper, and the peroxide compounds it creates, may directly alter structures on the viral envelope which are necessary for attachment to host cells. Peplomers, the viral glycoproteins protuberances which connect to host cell receptors are likely sites of ozone action. Alteration in peplomer integrity impairs attachment to host cellular membranes foiling viral attachment and penetration.
• Introduction of ozone into the serum portion of whole blood induces the formation of lipid and protein peroxides. While these peroxides are not toxic to the host in quantities produced by ozone therapy, they nevertheless possess oxidizing properties of their own which persist in the bloodstream for several hours. Peroxides created by ozone administration show long-term antiviral effects which serve to further reduce viral load. This factor may explain in part the reason for the fact that ozonated blood in the amount processed in usual treatment protocols is able to reduce viral load values in the total blood volume.
• Immunological effects of ozone have been documented. Cytokines are proteins manufactured by several different types of cells which regulate the functions of other cells. Mostly released by leucocytes, they are important in mobilizing the immune response. It has been found that ozone induces the release of cytokines which in turn activate a spectrum of immune cells. This is likely to constitute a significant avenue for the reduction of circulating virions.
• Ozone action on viral particles in infected blood yield several possible outcomes. One outcome is the modification of virions so that they remain structurally grossly intact yet sufficiently dysfunctional as to be nonpathogenic. This attenuation of viral particle functionality through slight modifications of the viral envelope, and possibly the viral genome itself, modifies pathogenicity and allows the host to increase the sophistication of its immune response. The creation of dysfunctional viruses by ozone offers unique therapeutic possibilities. In view of the fact that so many mutational variants exist in any one afflicted individual, the creation of an antigenic spectrum of crippled virions could provide for a unique host-specific stimulation of the immune system, thus designing what may be called a host-specific autovaccine.

Conclusion and Commentaries on Ozone Therapy and Research

In view of these considerations, it is evident that ozone presents fascinating opportunities for experimental and clinical research. Of all the antiviral agents known, none appear to offer the unique features of ozone, including its potent oxidizing power, and its gaseous nature. Indeed, as a gas, ozone is incomparably able to penetrate fluids with great ease and rapidity, delivering its biochemical actions almost instantaneously.

Research is needed to determine the effective range of ozone administration for the inactivation of each viral strain within all viral families. While this has been done for some members of major viral families, e.g., HIV and Hepatitis C, this know