Sign-up/Member Login                                 Microcard2u on Facebook
Loading
Bookmark and Share

bizarre topic gallery header
Topic Gallery Main Page   /    MyTopic

Contents: Proof Of Ozone ,


Air Cleaning Product Page

(INTRODUCTION),     (WHY OZONE IS A DRUG)   (METHODS OF OZONE GENERATION AND ADMINISTRATION) (THE EFFECTS OF OZONE ON PATHOGENS),     (CUTANEOUS PHYSIOLOGICAL EFFECTS OF OZONE),     (EXTERNAL MEDICAL CONDITIONS BENEFITED BY OZONE THERAPY),     (INFECTED WOUNDS), (POORLY HEALING WOUNDS),     (DECUBITUS ULCERS),    (CIRCULATORY DISORDERS),     (LYMPHATIC DISEASES),     (FUNGAL SKIN INFECTIONS),     (BURNS),     (GENERAL VIRAL CONDITIONS WITH REFERENCE TO CUTANEOUS AFFLICTIONS),     (NAIL AFFLICTIONS),    (RADIODERMATITIS),     (FROSTBITE),     (ADVANTAGES OF TOPICAL OZONE THERAPY),     (The salient advantages of topical ozone therapy include:),     (CONCLUSIONS),     (BIBLIOGRAPHY - Ozone),     (Possible Mechanisms of Viral Inactivation by Ozone),     (Comentaries on Prion Diseases and Ozone),     (BIBLIOGRAPHY - Comentaries on Prion Diseases and Ozone),     (Diabetic Leg Ulcers: A Missing Ingredient in Their Treatment and Management),     (Factors contributing to skin lesions in diabetes:),     (Circulatory impairment),     (Neuropathy),     (Mechanical stress),     (Ozone),    (Ozone as a drug),     (Ozone generation and administration),     (Ozone's actions on wound pathogens),     (Fungi),     (Protozoa),    (Ozone's cutaneous physiological effects),     (Diabetic skin conditions benefited by ozone therapy:),     (Diabetic leg ulcers),    (Gangrene),     (The practice of external ozone therapy in diabetic skin lesions),     (Advantages of topical ozone therapy in diabetes),    (Disadvantages of topical ozone therapy in diabetes),     (Conclusions),     (Suggested Reading and References),     (Hepatitis C and Ozone Therapy),    (Abstract),      (Ozone: Possible mechanisms of anti-viral action),      (Summary),(BIBLIOGRAPHY - Hepatitis C and Ozone Therapy),    (A Virology Primer: With Special Reference to Ozone),     (The Nature of Viruses),    (Components of Viruses),    (Major Enveloped Viruses Include the Following Families:),     (Major Non-Enveloped (Naked) Viruses:),    (The Viral Life Cycle),    (Commentaries on the Evolution of Viruses),     (Principles of Viral Inactivation),     (The viral culling effects of ozone in infected blood may recruit the following mechanisms:),     (Conclusion and Commentaries on Ozone Therapy and Research),     (Bibliography - A Virology Primer: With Special Reference to Ozone).

Proof of ozone>

From http://www.ozonehospital.com

The utilization of ozone for external medical applications

by Gerard V. Sunnen, M.D.
May 1998


INTRODUCTION

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.

back

WHY OZONE IS A DRUG

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.

back

METHODS OF OZONE GENERATION AND ADMINISTRATION

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.

back

THE EFFECTS OF OZONE ON PATHOGENS

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.

back

CUTANEOUS PHYSIOLOGICAL EFFECTS OF OZONE

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.

back

EXTERNAL MEDICAL CONDITIONS BENEFITED BY OZONE THERAPY

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:

INFECTED WOUNDS

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.

back

POORLY HEALING WOUNDS

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).

back

DECUBITUS ULCERS

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.

back

CIRCULATORY DISORDERS

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.

back

LYMPHATIC DISEASES

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.

back

FUNGAL SKIN INFECTIONS

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.

back

BURNS

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.

back

GENERAL VIRAL CONDITIONS WITH REFERENCE TO CUTANEOUS AFFLICTIONS

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.

back

NAIL AFFLICTIONS

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.

back

RADIODERMATITIS

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.

back

FROSTBITE

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.

back

ADVANTAGES OF TOPICAL OZONE THERAPY

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.

back

The salient advantages of topical ozone therapy include:

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.

back

CONCLUSIONS

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.

back

BIBLIOGRAPHY - Ozone

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

back

Possible Mechanisms
of Viral Inactivation by Ozone

by Gerard V. Sunnen, M.D.


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.

Gerard 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.

back

Comentaries on Prion Diseases and Ozone

by Gerard V. Sunnen, M.D.


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.

 

http://www.ozonehospital.com/prion.ilus340x312.gif

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.

back

BIBLIOGRAPHY - Comentaries on Prion Diseases and Ozone

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

back

Diabetic Leg Ulcers: A Missing Ingredient
in Their Treatment and Management

by Gerard V. Sunnen, M.D.
March 2007


Introduction

Diabetes is a disorder of metabolism and of the circulation. Chronic metabolic irregularities linked to poor circulatory perfusion and nerve damage can affect a number of organ systems, including skin tissues. In this article, the focus is on factors in diabetes that can contribute to dermal breakdown, ulceration, and infection. Most importantly, it proposes a treatment modality, which, backed by solid experimental, and clinical data cumulated worldwide, shows great promise in the management of diabetes-related skin lesions.

The conditions surveyed include infected wounds, skin ulcers and gangrene. These wounds, in the context of diabetes, are notoriously difficult to resolve. Healing resistance is thus a well-recognized element of frustration in their clinical care.

In most of the above conditions, multiple factors play into healing resistance. Among them are circulatory impairments, neurological deficits, tissue injury, and immunological compromise. A central factor is the proliferation of infectious microorganisms that, by the variety of their families, their toxin-producing capacities, and their resistance to antibiotics, offer daunting obstacles to standard treatment regimens.

Approximately 15% of the estimated 20 million Americans afflicted with diabetes mellitus develop lower leg skin ulcers. Of those patients, 20% will eventually require amputations. Diabetes mellitus is the leading cause of nontraumatic lower extremity amputation in the United States (LeRoith 2003).

back

Factors contributing to skin lesions in diabetes:

Circulatory impairment

Arteries and arterioles in chronic diabetes are prone to plaque buildup (Tesfaye 2005). The precise reason for this phenomenon is still elusive, yet it is well documented that Type II non-insulin dependent diabetes is linked to abnormal blood lipid profiles known as diabetic dyslipidemia (Goldberg 2004). Low-density lipoproteins particles are smaller in size and thus more apt to adhere to vessel walls, resulting in progressive vascular occlusion (Beckman 2002; Renard 2004). Lowered oxygen and nutrient supplies stress tissue resilience and impair recovery from injury (Chapnick 1996).

back

Neuropathy

Poorly controlled diabetes is correlated with peripheral nerve dysfunction. The mechanisms of diabetic injury to neurons are poorly understood. Higher blood glucose level seem to promote oxidative stress in neurons, but much more complex mechanisms are implicated (Tomlinson 2002).

Diabetic neuropathy can involve motor, sensory, and autonomic system neurons. Sensory neuron malfunction is translated as loss of feeling, reflex loss, problems with limb position sense, tingling (paresthesias) and pain. Motor impairment shows as muscle weakness. Autonomic neuropathy alters local circulation (Boulton 2004, Bensal 2006).

back

Mechanical stress

Chronic and repeating pressure on the skin compresses dermal arterioles, inhibiting tissue perfusion. Tissue weakness leads to ulceration. Ulcers are fertile ground for pathogenic microorganisms, and surrounding tissues become prone to cellulitis. At times, the ulcer crater reaches the underlying bone, initiating osteomyelitis (Boulton 2000).

back

Ozone

The oxygen atom exists in nature in several forms: (1) As a free atomic particle, singlet oxygen (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 (03 g 3/2 02 + 143 KJ/mole). It has a bond angle of 127� � 3�, resonates among several hybrid forms, is distinctly blue as a gas, and dark blue as a solid. (4) 04, a very unstable, rare, nonmagnetic pale blue gas readily breaks down into two molecules of oxygen.

Ozone, as a triatomic configuration of oxygen, possesses supreme oxidizing power derived from its marked tropism for extracting electrons from other molecules, simultaneously releasing one of its own oxygen atoms in the process.

back

Ozone as a drug

Ozone's capacity for inactivating microorganisms has been increasingly appreciated since the turn of the last century (Viebahn 1999). In the past few decades, ozone's action against bacteria, viruses and fungi has sparked keen interest for its use, not only for purifying water supplies, but also for medical objectives.

Ozone/oxygen mixtures exert significant antimicrobial activity. As with many medications, however, ozone has a range of action that, in the terminology of pharmacokinetics, is referred to as a therapeutic window (Bocci 2005). Indeed, ozone applied in concentrations that are too low, has little therapeutic effect. Applied externally in high concentrations, ozone may become irritating and tissue-toxic.

Due to ozone's demarcated therapeutic range, ozone concentrations administered to the patient need to be carefully calibrated and controlled. Optimally therapeutic ozone/oxygen mixtures require state of the art quantitative (dosage, concentration), as well as qualitative (purity) controls currently available in contemporary ozone generation technologies, all predicated upon the evaluation of the lesions under treatment.

back

Ozone generation and administration

Ozone is a gas with a half-life of approximately one hour at room temperature. Medical ozone generation and delivery systems therefore require that ozone be created at the moment it is to be administered. Ozone, in this sense is not a drug that has a shelf life enabling it to be kept for long periods of time.

Ozone is created by applying energy to oxygen. The oxygen source should be pure and devoid of nitrogen or other impurities. The presence of too much nitrogen favors the production of tissue-toxic nitrogen oxides.

Importantly, the humidity level of the ozone/oxygen mixture enters into the treatment protocol. Indeed, in certain wounds, humidity added to the ozone/oxygen mixture, markedly enhances therapeutic results.

back

Ozone's actions on wound pathogens

Bacteria fare poorly when exposed to ozone, a fact appreciated since the 19th century (Viebahn 1999). Ozone is a strong germicide needing only micrograms per liter for measurable action. At a concentration of 1 mg per liter of water at 1�C, ozone rapidly inactivates coliform bacteria, staphylococcus aureus, and Aeromonas hydrophilia (Lohr 1984). The inactivation rate for E. coli, takes place in relatively small concentrations of ozone, and is influenced by pH and temperature (Ivanova 1983).

At dosage concentrations used in external therapy, ozone essentially inactivates all bacterial species. This holds true for oxygen-dependent aerobic organisms, for oxygen-independent anaerobic bacteria associated with gangrene, and for facultative species that can function with or without oxygen. Spores and cysts are neutralized as well (Ishizaki 1986, Langlais 1986). Spores of Bacillus cereus and Bacillus megaterium are susceptible to ozone exposure (Broadwater 1973). Ozone's universal antibacterial action makes it an agent of choice in the management of wound infections colonized by bacterial species belonging to diverse groups.

An incomplete list of bacterial families susceptible to ozone inactivation includes the Enterobacteriaceae, a large group whose natural habitat is the intestinal tract of mammals. These Gram-negative organisms include Escherichia coli, Salmonella, Enterobacter, Shigella, Klebsiella, Serratia, and Proteus. Other ozone-sensitive bacterial species include Streptococci, Staphylococci, Legionella, Pseudomonas, Yersinia, Campylobacteri, and Mycobacteria (Dyas 1983, Broadwater 1973).

The cell envelopes of bacteria are composed of intricate multilayers. Covering the bacterial cytoplasm to form the innermost layer of the envelope is the cytoplasmic membrane, made of phospholipids and proteins. Next, a polymeric layer built with giant peptidoglycan molecules provides bacteria with a stable architecture. In Gram-positive organisms, the pepticoglycan shell is thick and rigid. By contrast, Gram-negative bacteria possess a thin pepticoglycan lamella on which is superimposed an outer membrane made of lipoproteins and lipopolysaccharides. In acid-fast bacteria, such as Mycobacterium, up to one half of the capsule is formed of complex lipids (Parish 2005, Hogg 2005).

The most cited explanation for ozone's bactericidal effects centers on disruption of cell membrane integrity through oxidation of its phospholipids and lipoproteins. There is evidence for interaction with proteins as well (Mudd 1969). In one study exploring the effect of ozone on E. coli, evidence was found for ozone's penetration through the cell membrane, breaking the closed circular plasmid DNA, which would presumably diminish the efficiency of bacterial procreation (Ishizaki 1987).

back

Fungi

Fungi are frequent inhabitants of chronically infected wounds. One study (Moussa 1999) found colonization by Candida and Aspergillus. Fungal organisms neutralized by ozone exposure include Candida, Aspergillus, Histoplasma, Actinomycoses, and Cryptococcus. The multilayered cell walls of fungi, composed of carbohydrates, proteins and glycoproteins, contain many disulfide bonds sensitive to ozone oxidation.

back

Protozoa

Protozoan organisms are often found in chronically infected wounds. Species disrupted by ozone include Giardia, Cryptosporidium, and free-living amoebas, including Acanthamoeba, Hartmonella, and Negleria. Several authors have demonstrated ozone�s capacity to penetrate through the walls of Giardia cysts causing fatal structural damage (Widmer 2002, Wickramanayake 1984).

back

Ozone's cutaneous physiological effects

Oxygen has long been established as beneficial in many pathological conditions, forming the basis for the use of hyperbaric oxygen treatment for carbon monoxide poisoning, decompression sickness, gas gangrene and stroke, among others. Oxygen under pressure, applied to infected tissues, inhibits the proliferation of anaerobic bacteria and stimulates local circulation (Wunderlich 2000).

Ozone, when added to oxygen, however, has properties that clearly transcend oxygen administration alone. The two properties invoked are:

1.     Ozone's extremely broad range of antipathogenic action and,

2.     The vasodilation of arterioles promoting tissue oxygenation and the delivery of nutrients and immunological factors to compromised tissues; and the vasodilation of veins, increasing venous outflow and the removal of toxins.

back

Diabetic skin conditions benefited by ozone therapy:

Wounds with a potential for infection

This category addresses wounds that are not yet infected but have a high probability for eventual infection. Post-surgical wounds, injuries such as abrasions, contusions and lacerations are salient examples.

The use of topical ozone therapy in these cases may be solely preventive, aimed at inhibiting the proliferation of potentially infective organisms. Preventative topical ozone therapy may thus stave off the development of potentially disastrous infectious complications.

Poorly healing wounds

Wounds healing in an indolent manner are apt to regress if treatment continuity is interrupted.

In these wounds, anaerobic bacteria - bacteria that do not need oxygen for their growth (e.g., Bacteroides, Clostridium) - may be active at deeper levels of the dermis, insulated from the influence of oxygen. While anaerobic bacteria are responsible for many devastating infections including gas gangrene, aerobic bacteria normally found on skin surfaces such as Staphylococcus epidermis, Corynebacteria, and Propionobacteria, given propitious circumstances, are capable of remarkable aggressive infectivity.

back

Diabetic leg ulcers

Diabetic ulceration is accelerated by poor circulation and neuropathy. One study (Anandi 2004) reported bacterial culture results for 107 patients with diabetic foot lesions. They included E. coli, Klebsiella, Pseudomonas, Proteus, Enterobacter, Clostridium perfringens, Bacteroides, Prevotella, and Peptostreptococcus.

The treatment of diabetic ulcers requires a multidisciplinary approach, including surgical, topical, and systemic interventions when indicated (Cavanagh 2005, Kruse 2006). Topical antibiotics often fail to penetrate far enough into the wound and frequently cause secondary dermatitis and allergy in their own right (De Groot 1994). For this reason, they are not generally recommended. Systemic antibiotics, prescribed for infections transgressing ulcer borders, can only address a portion of the spectrum of microorganisms cultured from such wounds. Bacterial resistance is common (e.g., �-lactam antibiotic resistance, as in methicillin-resistant staphylococcus).

Ozone applications in diabetic ulcers provide essential dual functions of topical broad-spectrum coverage and circulatory stimulation. In addition, ozone, via multiple serial applications and higher dose ranges, is able to further its penetration into deeper tissue layers where anaerobic bacteria are apt to reside.

back

Gangrene

Gas gangrene, also known as necrotizing fascitis, myositis, and myonecrosis is feared because of its rapid evolution leading to the galloping breakdown of affected tissues (Chapnick 1996, Falanga 2002)).

Several bacterial species are implicated in this process, the most common being Clostridium and toxin-producing Group A Streptococcus families. Other bacterial species implicated in gas gangrene include E. coli, Proteus, Staphylococcus, Vibrio, Bacteriodes, and Fusiforms (Caballero 1998). Gas gangrene may become a fatal complication of diabetic and decubitus ulcers.

Anaerobic and facultative bacteria feed on sugars and glycogen, produce lactic acid, and gases such as methane, carbon dioxide, and hydrogen. Their life threatening toxins cause severe tissue breakdown, hemolysis, renal failure, and shock.

These impressively destructive wounds demand emergency ozone application as an important adjunct to their multidisciplinary interventions.

back

The practice of external ozone therapy in diabetic skin lesions

In every case, an individual assessment has to be made relative to the skin lesion under treatment. Noted in this evaluation are the size (diameter and depth) of the lesion, and in deeper lesions, the involvement of dermal tissues, ligaments, muscle and bone. Also, the presence of purulence and necrosis, the relative health of surrounding tissues, and adjacent circulatory competence.

Ozone therapy is always individualized to incorporate these clinical observations. Accordingly, ozone concentrations are adjusted, as are lengths and frequencies of treatment, all recalibrated as treatment progresses.

In the practice of external ozone application, a specially designed ozone-resistant envelope is used to enclose the area being treated. A precise fitting of the envelope is needed in order to ensure a constant ozone/oxygen concentration within the envelope milieu and a proper containment of the gas. Ozone will thus be prevented from escaping into the ambient environment, reducing respiratory exposure to treating personnel.

The ozone concentrations prescribed during the course of treatment, the duration and frequency of individual sessions, and the lengths of the overall course of therapy are all predicated upon the evolution of the specific medical condition under treatment. In extensive wet ulcers and burns, for example, initial topical ozone concentrations need to be low in order to prevent excessive systemic ozone absorption. With gradual epitheliazation of the ulcer wound, applied ozone concentrations will require corresponding adjustments.

back

Advantages of topical ozone therapy in diabetes

1.     The ease of administration of this therapy. Once the principles of ozone dynamics and the art of adapting ozone dosages and treatment protocols are mastered by the clinician, topical oxygen/ozone therapy can safely be applied to a broad range of diabetes-related afflictions.

2.     Ozone is an effective antagonist to an enormous range of pathogenic organisms. In this regard, ozone cannot be equaled. It inactivates aerobic, facultative, and anaerobic bacterial organisms, a wide spectrum of viruses, and a comprehensive range of fungal and protozoan pathogens. To replicate this therapeutic action, ulcerative conditions would have to be treated with an assortment of various systemic antibiotic agents. In the context of accepted contemporary medical practice, this is not feasible.

3.     External ozone therapy, applied in a timely fashion, may obviate the need for systemic antipathogen therapy, thus saving the patient from all the side effects and organ stresses this option entails. External ozone is both a preventive, acute care, and chronic care therapeutic agent.

4.     External ozone application to superficial tissues whose blood supply is reduced enhances tissue blood and oxygen perfusion.

5.     There is evidence that ozone, via its oxidizing properties, inactivates bacterial toxins. Toxins, whose function is to destroy tissues, provide bacteria with colonizing advantage.

6.     Ozone exerts its anti pan-pathogenic actions through entirely different mechanisms than conventional antibiotic agents. The latter must be constantly upgraded to surmount pathogen resistance and mutational change. Ozone, on the other hand, presents a direct and powerful oxidative challenge that any and all pathogens are incapable of circumventing.

7.     Externally applied ozone/oxygen mixtures are entirely compatible with systemically administered antibiotics, as they are with debridement and other local wound care procedures.

back

Disadvantages of topical ozone therapy in diabetes

1.     Ozone/oxygen mixtures are not transportable and need to be created at the site and time of administration.

2.     Ozone/oxygen mixtures need to be administered serially in diabetic wounds. This may translate, in many circumstances, to daily applications until the lesion resolves.

3.     Ozone/oxygen mixtures, applied externally, have limited penetrability. While they possess panpathogenic power on ulcer surfaces, their therapeutic action has limited range at greater depths of ulcer boundaries.

back

Conclusions

Topical ozone/oxygen therapy has shown effectiveness and safety in healing diabetic skin afflictions. In this article, the following are cited: Wounds with potential for infection, infected wounds, poorly healing wounds, diabetic leg ulcers, decubitus ulcers and gangrene.

Ozone possesses unique physico-chemical attributes enabling it to exert potent antipathogenic activity. Applied to the adjunctive treatment and management of diabetic leg lesions, ozone can tip the balance from chronic failure to resolution. There is one crucial element missing from contemporary therapeutic regimens for diabetic skin lesions: Ozone

back

Suggested Reading and References

Ackey D, Walton TE. Liquid-phase study of ozone inactivation of Venezuelan Equine Encephalomyelitis virus. Appl Environ Microbiol 1985; 50: 882-886

Albrant DH. Management of foot ulcers in patients with diabetes. J Am Pharm Assoc 2000; 40(4): 467-474

Anandi C, Alaguraja D, Natarajan V et al. Bacteriology of diabetic foot lesions. Indian J Med Microbiol 2004; 22: 175-178

Armstrong. Infectious Diseases, First Ed. Mosby, Philadelphia, 2000

Beckman JA, Creager MA, Libby P. Diabetes and Atherosclerosis. JAMA 2002 May 15; 287:2570-2581

Bensal V, Kalita J, Misra UK. Diabetic neuropathy. Postgraduate Medical J 2006; 82:95-100

Berger J, Blum D, Bourdette, DN, Corey-Bloom J. Adult Neurology. Blackwell Publishing, 2005

Bocci V. Oxygen-Ozone Therapy: A Critical Evaluation. Kluwer Academic Publishers, Dordrecht, 2002

Bocci V. Biological and clinical effects of ozone. Br J Biomed Sci 1999 Jan; 56(4): 270-279

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

Boulton AJ. The diabetic foot: a global view. Diabetes Metab Res Rev 2000; 16 (suppl 1): 2-5

Boulton AJ, Malik RA, Arezzo JC et al. Diabetic somatic neuropathies. Diabetes Care 2004; 27:1458-1486

Broadwater WT, Hoehn RC, King PH. Sensitivity of three selected bacterial species to ozone. Applied microbiology 1973 Sept; 26(3): 391-393

Buckley RD, Hackney JD, Clarck K, Posin C. Ozone and human blood. Archives of Environmental Health 1975; 30:40-43

Caballero E, Frykberg RG. Diabetic foot infections. J Foot Ankle Surg 1998; 37:248-255

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

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

Cavanagh PR, Lipsky BA, Bradbury AW et al., Treatment of diabetic foot ulcers. The Lancet 2005 Nov 12; 366(9498): 1725-1735

Champion RH, Burton JL, Ebling FJ. Textbook of Dermatology. Blackwell Scientific Publications, Oxford, 1992

Chapnick EK, Abter E. Necrotizing soft-tissue infections. Infectious Disease Clinics of North America 1996; 10(4): 835-843

Clark M, Price PE. Is wound healing a true science or a clinical art? The Lancet 2004 Oct 16; 364 (9443): 1388-1389

Dailey JF. Blood. Medical Consulting Group, Arlington MA, 1998

De Groot AC, Weyland WJ, Nater JP. Unwanted Effects of Cosmetics and Drugs Used in Dermatology, Elsevier, Amsterdam, 1994

Dyas A, Boughton B, Das B. Ozone killing action against bacterial and fungal species. Journal of Clinical Pathology 1983; 36(10): 1102-1104

Epstein E. Common Skin Disorders, Saunders, Philadelphia, 1994

Evans AS, Kaslow RA (Eds). Viral infections of humans: Epidemiology and control. Plenum, New York, 1997

Falanga V, Phillips TJ, Harding KG et al. Text Atlas of Wound Management. Taylor and Francis, 2002

Goldberg IJ. Why does diabetes increase atherosclerosis: I don�t know. J Clin Investigation 2004 Sept 1; 114(5): 613-615

Gunther-Schlegel H. General Microbiology. Cambridge University Press, 2003

Harakeh M, Butler MJ. Factors influencing the ozone inactivation of enteric viruses in effluent. Ozone: Science and Engineering 1985; 6:235-243

Hogg S. Essential Microbiology. Wiley, New York, 2005

Ishizaki K, Shinriki N, Matsuyama N. Inactivation of Bacillus spores by gaseous ozone. J Applied Bacteriology 1986; 60(1): 67-72

shizaki K, Sawadaishi D, Muira K, Shinriki N. Effect of ozone on plasmid DNA of Escherichia coli in situ. Water Research 1987; 21(7): 828-823

Ivanova O, Bogdanov M, Miura K, Shinriki N. Ozone inactivation of enteroviruses in sewage. Vopr Virusol 1983; 0(6): 693-698

Jeffcoate WJ, Harding KG. Diabetic foot ulcers. The Lancet 2003 May 03; 361 (9368): 1545-1551

Knipe DM, Howley PM. Fundamental Virology, Fourth Edition. Lippincott Williams & Wilkins, New York, 2001

Komananapalli IR, Lau BH. Inactivation of bacteriophage lambda, Escherichia coli, and Candida albicans by ozone. Abst Gen Meet Am Soc Microbiol 1997 May; 97:457

Kruse I, Edelman S. Evaluation and treatment of diabetic foot ulcers. Clinical Diabetes 2006 Spring; 24(2): 91-93

LeRoith D, Olefsky JM, Taylor SI. Diabetes Mellitus: A Fundamental and Clinical Text. Lippincott Williams & Wilkins (3rd Ed), 2003

Lesher JL. An Atlas of Microbiology. Informa Healthcare, 2000

Lippincott Williams & Wilkins, Philadelphia, 2001

Langlais B, Perrine D. Action of ozone on trophozoites and free amoeba cysts, whether pathogenic or not. Ozone: Science and Engineering 1986; 8:187-198

Leland DS. Clinical Virology. Saunders, Philadelphia, 1996

Lesher JL. An Atlas of Microbiology of the Skin. Informa Health Care, 2000

London NJ, Donnely R. Ulcerated lower limb. BMJ 2000 June 10; 320 (7249): 1589-1591

Lorh A, Gratzek J. Bactericidal and paraciticidal effects of an activated air oxidant in a closed aquatic system. J Aquaric Aquat Sci. 1984; 4:1-8

McNair Scott DB, Lesher EC. Effect of ozone on survival and permeability of Escherichia coli. J Bacteriology 1963 Mar; 85(3): 567-576

Marhell EK, Voge M, John DT. Medical Parasitology. Saunders, Philadelphia, 1986

Max J. Antibodies kill by producing ozone. Science 2002 Nov 15; 298:1319

Menzel DB. Ozone: an overview of its toxicity in man and animals. J Toxicol Environ Health 1984; 13:183-204

Mousa HA. Fungal infection of burn wounds in patients with open and occlusive treatment methods. Eastern Mediterranean Health Journal 1999; 5(2): 333-336

Mudd JB, Leavitt R, Ongun A, McManus T. Reaction of ozone with amino acids and proteins. Atmos Environ 1969; 3:669-682

Murray PR (Ed). Manual of Clinical Microbiology. ASM Press, Washington DC, 1995

Olinescu R, Smith TL. Free Radicals in Medicine. Nova Science Publishers, Inc. Huntington, New York, 2002

O�Meara SM, Cullum NA, Majid M, Sheldon TA. Systematic review of antimicrobial agents used for chronic wounds. Brit. J Surgery 2001 Jan; 88(1): 4-21

Parish T. Mycobacterium: Molecular Microbiology of the Skin. Taylor & Francis, 2005

Poretsky L (Ed) Principles of Diabetes Mellitus, Springer, 2002

Razumovskii SD, Zaikov GE. Ozone and its reactions with organic compounds. Elsevier, Amsterdam, 1984

Renard CB, Kramer F, Johansson F et al. Diabetes and diabetes-associated lipid abnormalities have distinct effects on initiation and progression of atherosclerotic lesions. J Clin Investigations 2004; 114:659-668

Ropper AH, Brown RH. Adams and Victor's Principles of Neurology (8th Edition). McGraw-Hill, New York, 2005

Roy D, Wong PK, Engelbrecht RS, Chian ES. Mechanisms of enteroviral inactivation by ozone. Applied Environmental Microbiology 1981; 41:728-723

Ryan KJ (Ed). Medical Microbiology. Appleton & Lange, Norwalk, Connecticut, 1994

Sobsey MD. Inactivation of health-related microorganisms in water by disinfection processes. Water Science Technology 1989; 21(3): 179-195

Sunnen G. Ozone in medicine: Overview and future directions. Journal of Advancement in Medicine 1988; 1(3): 159-174

Sunnen G. Possible mechanisms of viral inactivation by ozone. Townsend Letter for Doctors 1994 Ap: 336

Tesfaye S, Chaturvedi N, Eaton SE et al. Vascular risk factors and diabetic neuropathy. NEJM 2005 Jan 27; 352(4): 341-350

Thanomsub B. Effects of ozone treatment on cell growth and ultrastructural changes in bacteria. J Gen Appl Microbiol 2002 Aug 01; 48(4): 193-199

 Tomlinson D, Bradley R, Harris R, Jenner P. Neurobiology of Diabetic Neuropathy. Academic Press, 2002

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

Vaughn JM, Chen YS, Novotny JF. Effects of ozone treatment on the infectivity of hepatitis A virus. Can J Microbiol 1990; 36: 557-560

Viebahn R. The Use of Ozone in Medicine. Odrei Publishers, Iffezheim, 1999

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

Wentworth P, McDunn JE, Wentworth AD, et al., Evidence for antibody-catalysed ozone formation in bacterial killing and inflammation. Science 13 Dec 2002; 298:2195-2199

Werkmeister H. Subatmospheric 02/03 treatment of therapy-resistant wounds and ulcerations. OzoNachrichten 1985; 4:53-59

White DO, Fenner FJ. Medical Virology, Fourth edition. Academic Press, New York, 1994

Wichramanayake G, Rubin A, Sproul O. Inactivation of Giardia lamblia cysts with ozone. Applied and Environ Microbiology 1984; 84:671-672

Widmer G, Clancy T, Ward H et al. Structural and biochemical alterations in Giardia lamblia cysts exposed to ozone. J Parasitology 2002; 88(6): 1100-1106

Wunderlich RP, Peters EJ, Lavery LA. Systemic hyperbaric oxygen therapy: lower extremity wound healing and the diabetic foot. Diabetes Care 2000; 23:1551-1555

Young SB, Setlow P. Mechanisms of bacillus subtilis spore resistance to and killing by aqueous ozone. J Applied Microbiology 2004 May; 96(5): 1133-1142

Yu BP. Cellular defenses against damage from reactive oxygen species. Physiological Reviews 1994 Jan; 74(1): 139-162

back

Hepatitis C and Ozone Therapy

by Gerard V. Sunnen, M.D.


February 2001

Abstract

Hepatitis C (HCV) is a global disease with an expanding incidence and prevalence base. Of massive public health importance, hepatitis C presents supremely challenging problems in view of its adaptability and its pathogenic capacity. The unique strategies that HCV utilizes to parasitize its host make it a formidable enemy and therapeutic interventions need considerable honing to counter its progress. Ozone, because of its special biological properties, has theoretical and practical attributes to make it a potent HCV inactivator.

History of the virus A form of hepatitis became recognized in the 1970's that resembled hepatitis B, serum hepatitis, and to a lesser extent hepatitis A, infectious hepatitis. It had, however, novel features, amongst them, a distinctive serological profile. In 1989, the genome of hepatitis C (HCV) was deciphered.

It is possible, by means of extrapolation from the genetic evolution of a virus, to approximate its age. Sequence genetic analysis points to the diversification of different HCV genotypes 200 to 400 years ago. Ancestors to these genotypes probably date back 100,000 or so years when viruses co-evolved with modern humans. Further analysis of genetic viral trees and Old and New World primates take the primordial forms of these viruses to primate speciation periods some 35 million years ago.

Today, in the context of human population growth, migration, and global travel, the hepatitis C virus has expanded its territories, geographically, and demographically. There is every indication that the evolution of this virus, in all its forms, is currently manifesting an accelerated phase.

Virion architecture and molecular biology The HCV particle is composed of a nucleocapsid containing its genome, an RNA single strand composed of approximately 9600 nucleotides, and its protein coating. The nucleocapsid is surrounded by an envelope which allows attachment and penetration into host cells. The genome encodes structural proteins designated as core (C), envelope 1 (E1), envelope 2 (E2), and P7 (unknown function), providing for virion architecture, and nonstructural proteins, mainly enzymes essential to the virion's life cycle, designated as NS2, NS3, NS4A, NS4B, NS5A, and NS5B. Proteases release structural and nonstructural proteins. Helicases unwind viral nucleic acid. Polymerases replicate RNA. Within this genome is located a hypervariable region implying an area of intensive genetic fluidity and mutational potential. HCV displays great genotypic flexibility which makes for sophisticated evasiveness to host defenses.

The nucleocapsid is surrounded by an envelope, a lipid bilayer associated with a union of carbohydrates and proteins, glycoproteins. Up to 60% of the lipid component of the envelope is phospholipid and the remainder is mostly cholesterol. It possesses projections called peplomers which facilitate attachment to host cells. One protein on peplomers of the HCV particle which is thought to be instrumental in the attachment process is designated CD-81.

The sequence of nucleotides within the HCV genome shows significant variations. Strains obtained from different parts of the world, for example, may differ substantially in their structural and nonstructural protein compositions. This has lead to a system of classification of the HCV family into 6 genotypes (1 to 6), and approximately 100 subtypes (designated a, b, c, ect.). Genotypes vary from each other by a factor of 30% over the entire genome. Subtypes vary by about 20%. Genotypes 1 to 3 have global distribution, while genotype 4 and 5 are found mainly in Africa, and 6 is distributed in Asia. Importantly, genotype and subtype differences have shown varying susceptibility to antiviral therapy.

Within any one afflicted individual, HCV particles do not show a homogeneous population. Instead, they function as a pool of genetically variant strains known as quasispecies. This is due to the high replication error inherent in the function of the polymerase enzymes. Herein lies one of the important armaments of HCV. Continuously generated genetic diversity gives it great advantage in negotiating and conquering immune defense and therapeutic strategies. Furthermore, the antigenic differences between genotypes may have implications regarding the proper evaluation and the therapeutic regimen of patients.

Viral life cycle A freely circulating virion enters a host cell by binding to a cell surface receptor. In the case of HCV the host cell is a hepatocyte. However, bone marrow, kidney cells, macrophages, lymphocytes, and granulocytes may also be trespassed.

Once cell entry is achieved, the virion sheds its envelope to commence its replication. It binds to cellular ribosomes and released viral polymerase begins the RNA replication cycle. Newly formed nucleocapsids continue their assembly with the acquisition of new envelopes by means of budding through membranes of the cell's endoplamic reticulum. Newly formed virions may number in the range of 10 billion daily. The average life span of virions is in the order of a few hours.

Virions are then released into the general blood and lymphatic circulation, ready to infect new cells, re-infect already diseased cells, or a new host, mainly through bodily fluid transmission pathways. HCV RNA, as measured by polymerase chain reaction (PCR) may show 10 million or more virions per ml. As little as 0.0001 ml of blood may be sufficient to impart infection. The evolution of hepatitis C is characterized by phases of accentuated viremia punctuated by periods of relative quiescence. The presence and timely detection of these viremic waves may offer novel therapeutic considerations.

Clinical and laboratory manifestations Hepatitis, from anyone of the several viruses capable of inducing liver inflammation, produce a spectrum of clinical and laboratory manifestations. Hepatitis C distinguishes itself by the low incidence of acute phases and by the high incidence of progression to chronicity. Acute hepatitis C progresses from exposure, to incubation, to pre-icteric, icteric, and convalescent phases. With an incubation period of about 6 weeks, the first and sometimes only symptoms include weakness, fatigue, indolence, headache, nausea, poor appetite, and vague abdominal pain. The pre-icteric period extends from the onset of symptoms to the appearance of jaundice, ranging usually from 2 to 12 days. The icteric phase corresponds to the declaration of jaundice and darkened urine. The convalescent phase is marked by the gradual disappearance of symptoms.

Chronic hepatitis C is characterized by the presence of HCV RNA and the elevation of liver enzymes for 6 months or longer. Patients may be asymptomatic, or at times suffer an acute exacerbation with a return of symptoms. Approximately 75% of acutely ill patients continue into a chronic phase evidenced by parameters of viral presence.

Hepatitis C can only be distinguished from other viral hepatic conditions by serological and virological determinations. Liver enzymes characteristically affected by HCV infection include serum alanine transfesferase (ALT), aspartate aminotransferase (AST), gamma- glutamyl transpeptidase (GGTP), and alkaline phosphatase; in addition, there may be abnormalities in bilirubin, serum albumin, prothrombin time, and platelet density.

Cirrhosis, a diffuse disruption of liver tissue architecture with regenerative nodules surrounded by fibrosis, is an important sequel to hepatitis C. Within 20 years post HCV infection 20 to 25% of patients will develop cirrhosis. Hepatic decompensation ensues with ascites as the salient marker.

Hepatocellular carcinoma, another notable outcome of HCV infection is present in approximately 5% of patients post infection. The presence of cirrhosis is central to its genesis. Although the mechanisms by which cirrhosis ushers carcinoma are unknown, it is likely that chronic inflammation and the sustained pressure of cellular regeneration play important roles.

Up to 10% of patients appear to have fully conquered the disease. HCV antibodies are undetectable, as is HCV RNA. Liver enzymes are fully normalized, but liver biopsy may show lingering areas of stagnant inflammation and spotty necrosis. It is thus possible for host immunocompetence to vanquish HCV infection and therapeutic strategies aim to assist the host immune system to achieve this goal.

Immunological response to the virus HCV particles are detected early in the infection, usually 1 to 2 weeks following exposure. Antibodies to HCV core, nonstructural, and envelope elements appear about 6 weeks after exposure. A broad range of cytokines are mobilized. Cellular immunity is activated with broad recruitment of neutrophils, natural killer (NK), macrophages, and CD4 and CD8 T helper cells.

Current and experimental treatment strategies As of this date the main treatment strategies for hepatitis C include interferon and ribavirin. Interferons are natural cellular products which activate macrophages, neutrophils and natural killer cells. There is controversy as to interferon's biological effects, be they mostly immunoregulatory or directly antiviral. Ribavirin is a guanosine analog that represses messenger RNA formation thus inhibiting the replication of many DNA and RNA viruses. It is, however, mutagenic to mammalian cells. Ribavirin and interferon have significant medical and psychiatric side effects.

Treatment response is defined as undetectable viral load 6 months following therapy. Contemporary detection methods of quantitative HCV RNA determinations are capable of detecting approximately 1000 viral copies per serum ml.

Resistance to antiviral therapies is a particularly vexing problem in anti HCV treatment. Novel and experimental antiviral compounds include inhibitors of protease, polymerase and helicase.

Vaccine development needs to take into account HCV's antigenic rainbow and its high mutability. High mutation rates in this condition implies a dauntingly diverse and variable array of viral antigenic components. It is estimated, for example, that HCV mutates significantly in its own host approximately a thousand times a year. This implies that within any one afflicted individual there exists an awesomely large array of viral quasispecies, which in turn creates commensurate difficulties in the creation of effective vaccines.

Ozone: Physical and physiological properties Ozone (O3) is a naturally occurring configuration of three oxygen atoms. With a molecular weight of 48, the ozone molecule contains a large excess of energy. It has a bond angle of 127� and resonates among several forms. At room temperature, ozone has a half life of about one hour, reverting to oxygen. A powerful oxidant, ozone has unique biological properties which are being investigated for applications in various medical fields. Basic research on ozone's biological dynamics have centered upon its effects on blood cellular elements (erythrocytes, leucocytes, and platelets), and to its serum components (proteins, lipoproteins, lipids, carbohydrates, electrolytes). Administrating increaing dosages of ozone to whole blood shows that beyond a certain threshold there is a rise in the rate of hemolysis. This threshold, depending upon various parameters, begins to be reached at 40 to 60 micrograms per milliliter, and becomes significant when higher levels are attained. Precise ozone dosing capacity is therefore essential in clinical practice and research.

Leucocytes show good resistance to ozone because they have enzymes which protect them from oxidative stress. These enzymes include superoxide dismutase, glutathione, and catalase. Research has shown that platelets also maintain their integrity after ozone administration. In ozone therapy, the doses applied to blood are gauged to avoid disruption of its cellular elements. Serum components remain viable during ozone therapy. Lipid and protein peroxides, produced in small amounts by ozonation, have demonstrable antiviral properties. Interestingly, ozone tends to stimulate leucocyte function and cytokine production. Ozone increases the oxygen saturation (p02) in erythrocytes and enhances their pliability so that capillary circulation is facilitated.

Ozone: Antiviral properties Recently, there has surged renewed interest in the potential of ozone for viral inactivation. It has long been established that ozone neutralizes bacteria, viruses, and fungi in aqueous media. This has prompted the creation of water purification processing plants in many major municipalities worldwide.

Ozone's antiviral properties may also be applied to the treatment of biological fluids, albeit in technologically and physiologically appropriate ways. Indeed, it is noted that ozone, administered in such dosages designed to respect the integrity of blood's cellular and constituent elements, is capable of inactivating a spectrum of viral families.

Some viruses are much more susceptible to ozone's action than others. It has been found that lipid-enveloped viruses are the most sensitive. This group includes, amongst others, HCV, Herpes 1 and 2, Cytomegalus, HIV1 and 2.

The envelopes of viruses provide for intricate cell attachment, penetration, and cell exit strategies. Peplomers, finely tuned to adjust to changing receptors on a variety of host cells, constantly elaborate new glycoproteins under the direction of E1 and E2 portions of the HCV genome. Envelopes are fragile. They can be disrupted by ozone and its by-products.

In HCV, viral load appears to be a major factor in the invasiveness and virulence of the disease process. Preliminary research has shown that reduction of viral load in Hepatitis C by means of ozone therapy can significantly normalize hepatic enzymes and improve measures of global patient health. Volunteers administered ozone therapy according to the method outlined below achieved a viral load reduction in the order of 5 log, or 99.9%, along with a normalization of liver enzyme levels.

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 hepatitis C, 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.

The aliquots of blood range from 50 ml. to 300 ml. Ozone dosages and treatment frequency vary according to treatment protocols. The reason aliquots of blood are treated and not, as one would propose, the entire blood volume, is that in the latter case the total ozone dosage administered would exceed toxic limits.

The average adult has 4 to 6 liters of blood, accounting for about 7% of body weight. How can the viral load reduction observed via ozone therapy be explained in the face of a technique that treats relatively small amount of blood, albeit serially?

back

Ozone: Possible mechanisms of anti-viral action

The viral culling effects of ozone in infected blood may recruit the following mechanisms:

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.

back

Summary

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 evolved impressive genetic diversity and mutational capacity to cope with the changing ecology of planetary life.

HCV has an extremely high rate of mutation and within any one individual there may exist millions of antigenic quasispecies. The disease process is marked by periods of viral quiescence alternating with viremic waves whereby billions of virions are poured into the blood and lymphatic reservoirs. Their astounding numbers stress the immune system relentlessly and produce an inexorable compromise in all parameters of its functioning.

Viral load reduction by means of ozone blood treatment alleviates immune system fatigue. Ozone-mediated viral culling may be achieved by anyone of a number of possible mechanisms. Direct virion denaturation, peplomer alteration, lipid and protein peroxide formation, cytokine induction, host pan-humoral activation, and host-specific autovaccine creation are suggested mechanisms. Due to the excess energy contained within the ozone molecule, it is theoretically likely that ozone, unlike antiviral options available today, will show effectiveness across the entire genotype and subtype spectrum.

Ozone embodies unique physico-chemical and biological properties which suggest an important role in the therapy of hepatitis C, either as a monotherapy, or as an adjunct to standard treatment regimens.

back

BIBLIOGRAPHY - Hepatitis C and Ozone Therapy

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

back

A Virology Primer: With Special Reference to Ozone

Revised October 2001

by Gerard V. Sunnen, M.D.


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.

back

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.


Size of microscopic entities and microscope resolution

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


http://www.ozonehospital.com/icosahedral337x446.gif

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


http://www.ozonehospital.com/helical338x466.gif

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.

back

Major Enveloped Viruses Include the Following Families:

Hepadnaviridae (from Hep: liver; dna: DNA)

Hepatitis B (HBV).

Retroviridae (from retro:reverse, to describe the reverse transciption allowing these viruses to make DNA from RNA)

 

HIV-1, Immunodeficiency syndrome

 

HIV-2, Immunodeficiency syndrome

Herpesviridae (herpes: to creep, describing creeping vesicular lesions)

Herpes simplex, types I and 2

 

Varicella-zoster: Chicken pox and shingles

Cytomegalovirus (from cyto: cell; megalo: large)

 

Epstein-Barr causes infectious mononucleosis; implicated in chronic fatigue syndrome

Filoviridae (filamentous viruses with hook shapes)

Ebola virus (Ebola river, Zaire): Hemorrhagic fever

Marburg (Marburg, Germany): Hemorrhagic fever

Orthomyxoviridae (Ortho: normal; myxo: mucus)

Influenza A and B

Paramyxoviridae (resembles Orthomyxoviridae)

Measles

Mumps

Parainfluenza

 

Respiratory syncytial virus: Flu-like syndrome

Coronaviridae (Capsid resembles a crown of thorns). Syndrome resembles flu.

Togaviridae (Toga: coat)

 

Rubella - German measles

 

Eastern and Western equine encephalitis. Transmitted by mosquitoes.

Rhabdoviridae (Rhabdos: rod)

Rabies

Bunyaviridae (Bunyamera, Uganda): Encephalitis

Flaviviridae (Flavus: yellow)

Hepatitis C

back

Major Non-Enveloped (Naked) Viruses:

Adenoviridae (from adenoid tissues)

Adenovirus: Respiratory infections, gastroanteritis, cystitis

Papovaviridae (from papilloma, polyoma, vacuolating

Polyomavirus, Papillomavirus (warts)

Picornaviridae (pico: small)

Poliovirus (polios: gray; myelitis: infection of the myelin in the spinal cord)

 

Echovirus (from E-enteric, C-cytopathogenic, H-human, 0-orphan): Aseptic meningitis

 

Coxsachie virus: Pharyngitis, aseptic meningitis, pericarditis, myocarditis

 

Hepatitis A: Infectious hepatitis

 

Rhinovirus (Rhino: nose): respiratory infection

Reoviridae (Respiratory, enteric, orphan)

Rotavirus (Rota: Wheel) - Severe diarrhea in infants and adults

Caliciviridae (Chalice-like): Gastroenteritis

back

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.

back

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.

back

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 .

back

The viral culling effects of ozone in infected blood may recruit the following mechanisms:

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.

back

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 knowledge applied to every main viral species will assist greatly in the precise methods for decontamination of mammalian products (serum, vaccines, etc.), and in the proper administration of therapeutic ozone in clinical situations. Research is needed, as well, to elucidate more precisely the mechanisms by which ozone decreases viral load.

Viral diseases are expanding worldwide. This is not a science fiction scenario but a regrettable fact. Old diseases are not only finding increased population reservoirs, but are also developing novel mechanisms of infectivity. Of special concern are emergent viruses, products of mutational creativity, never before encountered by humans, which are potentially devastating to health and life. Ozone has the unparalleled promise to find a major place in the armamentarium against these new plagues.

Viral load reduction by means of ozone blood treatment alleviates immune system fatigue. Due to the excess energy contained within the ozone molecule, it is theoretically likely that ozone, unlike antiviral options available today, will show effectiveness across the entire genotype and subtype spectrum. Ozone embodies unique physico-chemical and biological properties which suggest an important role in the therapy of a variety of viral conditions, most likely those mediated by lipid enveloped viruses, either as a monotherapy, or as an adjunct to standard treatment regimens.

back

Bibliography - A Virology Primer: With Special Reference to Ozone

Akey DH, Walton DE. Liquid phase study of ozone inactivation of Venezuelan equine encephalitis virus. Applied Environmental Microbiology 1985; 50: 882

Bocci V. Autohemotherapy after treatment of blood with ozone. A reappraisal. Journal of International Medical Research 1994 May-Jun; 22(3): 131-144

Bocci V. Ozonation of blood for the therapy of viral diseases and immunodeficiencies. A hypothesis. Medical Hypotheses 1992 Sept; 39(1): 30-34

Bocci V, et al. Studies on the biological effects of ozone: Generation of reactive oxygen species (ROS) after exposure of human blood to ozone. Journal of Biological Regulators and Homeostatic Agents 1998 July-Sept; 12(3): 67-75

Bolton DC, Zee YC, Osebold JW. The biological effects of ozone on representative members of five groups of animal viruses. Environmental Research 1982; 27: 476-484

Bolton DC, Tarkington BK, Zee YC, Osebold JW. An in vitro system for studying the effects of ozone on mammalian cell cultures and viruses. Environmental Research 1982; 27: 466-475

Buckley RD, Hackney JD, Clark K, Posin C. Ozone and human blood. Archives of Environmental Health 1975; 30: 40-43

Burleson GR, Murray TM, Pollard M. Inactivation of viruses and bacteria by ozone with and without sonication. Applied Microbiology 1975; 29: 340

Cann AJ. Principles of Molecular Virology. Academic Press, San Diego, 1997

Carpendale MT, Freeberg JK. Ozone inactivates HIV at noncytotoxic concentrations. Antiviral Research 1991; 16: 281-292

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

Evans AS, Kaslow RA (Eds) Viral Infections in Humans: Epidemiology and Control, Fourth Edition, Plenum, New York, 1997

Garber GE, Cameron DW, Hawley-Foss N, et al. The use of ozone-treated blood in the therapy of HIV infection and immune disease - a pilot study of safety and efficacy. AIDS 1991; 5: 981-984

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

Grob PJ. Hepatitis B: virus, pathogenesis, and treatment. Vaccine 1998; 16 Suppl S:11-16 Harrison TJ, Zuckerman AJ (Eds) The Molecular Medicine of Viral Hepatitis. Molecular Medical Science Series. John Wiley & Sons, New York, 1997

Kim CK, Gentile DM, Sproul OJ. Mechanism of ozone inactivation of bacteriophage f2. Applied Environmental Microbiology 1980; 39: 210-218

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

Leland DS. Clinical virology. Saunders, Philadelphia, 1996

Liang TJ, Hoofnagle JH (Eds) Hepatitis C. Academic Press, San Diego, 2000

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

Razumovskii SD, Zaikov GE. Ozone and its reactions with organic compounds. Elsevier, Amsterdam , 1984

Roy D, Wong PK, Engelbrecht RS, Chian ES. Mechanism of enteroviral inactivation by ozone. Applied Environmental Microbiology 1981; 41: 718-723

Sunnen GV. Ozone in medicine: Overview and future directions. Journal of Advancement in Medicine 1988; 1(3): 159-174

Turner BG, et al. Structural biology of HIV. Journal of Molecular Biology 1999 Jan; 285(1): 1-32

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, Heidelberg, 1994

Wells KH, Latino J, Gavalchin J, Poiesz BJ. Inactivation of human immunodeficiency virus Type I by ozone in vitro. Blood 1991; 78(7): 1182-1890

Yu BP. Cellular defenses against damage from reactive oxygen species. Physiological Reviews 1994 Jan; 74(1): 139-162  

Microcard2u, Bizarre Creative Consultants, Microshop2u Footer
Microcard2u Microcard2u
Chemicaboy
Microcard2u
Chemicaboy
Microcard2u Microcard2u 0162632281
Our Product Available At Store Below
  Product Video Comment To Microcard2u.com Charity Site
Topic Gallery MyTopic TalkTalk Forum Forex
 
Copyright © 2008 - 2017 Bizarre Creative Consultants. | Powered by Microcard Marketing | Privacy Statement