- Decrease in blood lactate concentration after maximal effort
- Increased pyruvate oxidation, thus increasing Acetil-Coa production
- Activation of cellular enzymal protection system
- Activation of the erythrocyte metabolis
- Increased 2,3-diphosphogliceric acid cycle
- Increased redox function of the mitochondrial respiratory chain.

'In a study on equine laminitis with twelve animals, the overall health situation of all of the horses improved substantially', says Kempchen.

Today about 60 per cent of the treated animals only suffer from minor ailments or are even free of complaints. Before they became candidates for participating in this study, veterinary euthanasia would have been the only alternative.

But the vast opportunities of hyerbaric therapy are not restricted to horses. The animal hospital in Wahlstedt has been very successful in treating dogs with ozone as well. Open fractures are often accompanied by wound healing disorders. In many of these cases, hyperbaric ozone therapy has been an efficient supportive measure to alleviate suffering and accelerate the healing process. 

NATO endorses joint U.S. / Canadian O3 Ozone oxygen research. 100% safe inactivation of HIV, Hepatitis, Herpes Virus in blood.

In a brief to the NATO Blood Committee' the Surgeon General of the Canadian Armed Forces reported on joint U.S. Canadian findings that within the sensitivity of the screening methods used, a 3-minute ozonation of serum spiked with one million HIV-1 particles per millilitre would achieve virtually 100% viral inactivation (loss of infectivity):

* N.A.T.O.: The NATO Blood Committee endorses joint U.S. / Canadian findings regarding O3 Ozone research.

* United States and Canada: 3-minute ozonation of serum spiked with one million HIV-1 particles per millilitre would achieve virtually 100% viral inactivation (loss of infectivity).

* Germany: 10 000+ units of human blood containing hepatitis virus were rendered safe through treatment with a mixture of ozone (O3) and oxygen (O2) for transfusion.

* Canada: Clinical trial of O3 Ozone in the treatment of AIDS patients (by means of phlebotomy, O3 ozonation of the blood sample and intramuscular reinoculation) ... showed no evidence of serious side effects.

* United States: Two teams of US virologists have used comparable gas-diffusion techniques to confirm the Canadian findings.

* United State and Europe: Findings at laboratories in North America and Europe have demonstrated that ozone has remarkable potency against disease factors in blood products.

* United States and Canada: Ozonized biologic fluids, in which there is a presumed abundance of superoxide, singlet oxygen, hydroxyl and peroxyl radicals and other highly reactive species, had both extracellular and intracellular virucidal properties.

Ref: http://mosao2.org/Article%20-%20O3/O3_HIV_NATO_Canada.html

Ref: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1490857/

Ozone was found to inactivate HIV-1 virions in a dose-dependent manner. Greater than 11 log inactivation was achieved within 2 hours at a concentration of 1,200 ppm ozone. The data indicate that the antiviral effects of ozone include viral particle disruption, reverse transcriptase inactivation, and/or a perturbation of the ability of the virus to bind to its receptor on target cells.

Ozone has potent anti-HIV-1 activity in cell culture media and factor VI11 preparations ... it is readily apparent that ozone may be of use in rendering factor VI11 and possibly other blood products, both proteinaceous and cellular, free of HIV-1 and other infectious agents. Ozone treatment offers promise as a means to inactivate human retroviruses in human body fluids and blood product preparations.

1DVM Clínica Veterinaria Dr. Vigna. Vía Primo Maggio, 34 40026, Imola, Italy. 
2 Ozone Research Center. POBox 6414, Havana, Cuba.


ivanvigna@tin.it
silvia.gra@infomed.sld.cu

ABSTRACT:

The satisfactory use of ozone therapy in the field of Ophthalmology has been already reported. For the therapy of the eye anterior chamber (above all the herpetic keratitis), in humans and Veterinary, the application of ozone injection, by subconjunctival way, the use of ozonized sunflower oil (OLEOZON) collyrium, in conjunction with ozone rectal insufflation, are well known.

Taking into account the OLEOZON great germicide power, the capacity to favor the cicatrization and the anti-inflammatory effect, the aim of this paper is to evaluate its efficacy applying, as unique medication, in cats affected by herpetic keratitis and chlamydiosis, associated to the midriatic in order to avoid the iris anterior synechia, as well as in corneal ulcer and in corneal ulcer treated with conjunctival flaps.

This study was performed in cats of different origin and age. Local treatment over the lesions was applied using OLEOZON collyrium, every 8 hours. Evaluation was performed every 72 h and every 4 h after the conjunctival flaps. The healing criteria were the inflammation disappearance and ulcer cicatrization.

In comparison with known pharmacological conventional therapies, the OLEOZON collyrium treatment was superior, more simple and more rapidly to achieve the results.

Only one contraindication of this medication was observed in 1 cat with herpetic symblepharon, due to the capacity of OLEOZON to increase fibroblasts. It is interesting to consider the good corneal cicatrization obtained with this treatment. No side effects were observed during the study

'In a study on equine laminitis with twelve animals, the overall health situation of all of the horses improved substantially', says Kempchen.

Today about 60 per cent of the treated animals only suffer from minor ailments or are even free of complaints. Before they became candidates for participating in this study, veterinary euthanasia would have been the only alternative.

But the vast opportunities of hyerbaric therapy are not restricted to horses. The animal hospital in Wahlstedt has been very successful in treating dogs with ozone as well. Open fractures are often accompanied by wound healing disorders. In many of these cases, hyperbaric ozone therapy has been an efficient supportive measure to alleviate suffering and accelerate the healing process. 

Ozone was found to inactivate HIV-1 virions in a dose-dependent manner. Greater than 11 log inactivation was achieved within 2 hours at a concentration of 1,200 ppm ozone. The data indicate that the antiviral effects of ozone include viral particle disruption, reverse transcriptase inactivation, and/or a perturbation of the ability of the virus to bind to its receptor on target cells.

Ozone has potent anti-HIV-1 activity in cell culture media and factor VI11 preparations ... it is readily apparent that ozone may be of use in rendering factor VI11 and possibly other blood products, both proteinaceous and cellular, free of HIV-1 and other infectious agents. Ozone treatment offers promise as a means to inactivate human retroviruses in human body fluids and blood product preparations.

Abstract

West Nile virus (WNV) poses a serious threat to public health, especially to the elderly and the immuno-compromised. Neither vaccines nor treatments are available for humans. Active hexose correlated compound (AHCC) is an extract ofLentinula edodes of the Basidiomycete family of fungi rich in α-glucans. In this study, we evaluated the effect of AHCC on host susceptibility in the murine model of WNV infection.

Mice orally administered with AHCC (600 mg/kg) every other day for 1 wk before and at d 1 and 3 postinfection were assessed using viremia levels, survival rate, and protective immunity. AHCC administration in young (6- to 8-wk-old) mice attenuated viremia and mortality following lethal WNV infection. WNV-specific IgM and IgG production and γδ T cell expansion were also enhanced in these mice. Aged (21- to 22-mo-old) mice were more susceptible to WNV infection than young mice, partially due to the dysfunction ofγδ T cell subsets.

AHCC administration in aged mice enhanced the protective Vγ1+ T cell response as well as WNV-specific IgG but not IgM antibodies production.

AHCC administration in aged mice attenuated viremia levels but led to no difference in mortality rate.

Overall, our data suggests that AHCC enhances protective host immune responses against WNV infection in young and aged mice. Dietary supplementation with AHCC may be potentially immunotherapeutic for WNV-susceptible populations.

http://jn.nutrition.org/content/early/2009/01/13/jn.108.100297

http://www.ahccresearch.com/testimonials.html

Cats:

OZ-PO-037: NEURO-PHYSIOLOGICAL AND BIOCHEMICAL EFFECTS OF OZONIZED SOLUTIONS APPLICATION IN EXPERIMENTS WITH CATS.

S.Kotov; A.Edeleva and A.Mochalov.
Nizhni Novgorod Medical State Academy. Russia.

In the mechanism of the biological action of ozone there are many obscure questions concerning its immediate influence on the nervous tissue. The aim of this research is to study, in experimental conditions, the influence of an ozonized solution containing middle therapeutic concentration of ozone in the somatosensory caused potential (SP), in the enzyme activity of the aerobic metabolism and in the contents of peroxidation lipid products in a somatosensory zone of the cortex in cats brain.

The registration of the caused potential was made on an open brain of 13 adult cats which received intraperitoneal nembutalo-chlorosoli narcosis. With this purpose an electrical boring rendered branches of the radial nerve in the area of the wished joint. Consecutive application of nonozonized and ozonized solution was carried out. In subsequent, the site of the brain was exposed to biochemical researches.

The ozonized solution causes a peak latence decrease of a negative phase in caused potentials to 85 %, from an initial level, in combination with a convertible short-term (10 min) augmentation of the positive phase amplitude in caused potential (26 %). Simultaneously, insignificant raises of the initial peroxidation lipid products were obtained in the researched site of the brain.

The enzymes activity of the respiratory chain does not change simultaneously with the appreciable intensifying of oxidoreductases, activity such as: lactat- dehydrogenases, piruvatdehydrogenases, succinatedehydrogenases.

The ozonized solution stimulates the aerobic glycolysis, raises functional neurones activity of the top cortical layers, not rendering any toxic effect on the excitability and conduction of the nervous tissue

Dogs:

OZ-P-073: O3 OZONE TREATMENT OF APPLICATION In The malignant NEOPLASMS IN DOGS.

T.Scherbatyuk, G.Poslov and V.Illarionov. 
Medical Academy, Nizhny Novgorod, Russia.

Dogs with malignant neoplasms Were Used In This Study. 

Ozone therapy by intravenous infusion WAS Applied Physiological saline solution of ozonized (100 - 200 mL, ozone concentration in the gas mixture 800 mcg / L) and by local application: intratumoral and peritumoral injection of Physiological saline solution ozonized. A cycle of Treatment Lasted from 5 to 15 days.

The results Indicated That, ozone therapy on malignant Animals with tumors, tumor metabolism Affected STI and stopped action on pathologic organism, Confirmed by the normalization of blood biochemical indices in tumor Treated animals.

Free-radical attack Caused by local action of ozone led to the Intensification of tumor cell respiration, on one hand, and to the suppression of the antioxidant defense system neoplasia, on the other hand. 

Cell death of uncertain part of tumor popullation finally occured, Which led to the DECREASE in the sizes and the necrotic blastoma Processes development. Surgical procedure, In This case, in order to necesario is clean the damage of the necrotic tissue Tumour mass.

Pigs:

O3 Ozone Therapy: a New Option in Veterinary Medicine.

Camps A.M. and Elías-Calles B.

The effect of ozonized sunflower oil (OLEOZON®) on tissue wounds performed by castration of Yorkshire race pigs was studied. The experiment was done after performing a wound in the scrotal region of fifteen pigs with 90 days old and sixty three pounds weight. They were divided into three groups (A, B and C) of five animals each. Group A - animals treated with placebo, once a day, topically, during five days. Group B - animals treated with OLEOZON®, once a day, topically, during five days. Group C - animals treated with OLEOZON®, twice a day, topically, during five days.

The results showed an important positive effect of OLEOZON® on wound healings, achieving the best results in group C in which the wound scarred more rapidly than in groups B and A, with significant differences (p < 0.05). Our data indicate that the topic application of OLEOZON® has a cicatrization and regenerating effects on dermic wounds in pigs.

Rodents:

OZ-P-062: A STUDY OF THE EFFECT OF O3 OZONE MUTAGENIC rectal insufflation ADMINISTERED BY IN RODENTS.

National Center for Laboratory Animal Production,
Ozone Research Center.

The reaction of ozone with lipids occurs exclusively at the level of carbon-carbon double bonds present in the polyunsaturated fatty acids, organic peroxides and generating ozonides, which in adequate amounts and controlled exert different biological actions, which give a set ozone therapeutic properties. 

In this work we studied the possible mutagenic effects of ozone applied rectally. We used analysis of bone marrow in NMRI mice by the micronucleus test and SPRD rats using the chromosome aberration test. Two concentrations were used ozone (50 and 75 mg / mL). As negative and positive controls were used oxygen and cyclophosphamide (40 mg / kg) respectively. 10 administrations were performed every 24 hours. At the end of treatment animals were sacrificed and processed for bone marrow preparations. 

The results show no evidence of cytotoxic or clastogenic (induction of chromosome aberrations) in the bone marrow of rodents treated.

Protective Effects of O3 Ozone Oxidative Preconditioning Against Liver Ischemia/Reperfusion Injury in Rats.

León O.S. and Ajamiech H.

Liver transplantation is a recommended therapy for end-stage liver disease and the demand for donor organs has surpassed the supply resulting in the death of thousands of patients. The damage by hepatic ischemia/reperfusion (I/R) is considered as the main responsible of organic failure after transplantation. On the other hand, 30% of transplants still fail by acute or chronic rejection within five years.

The mechanisms of acute liver damage following I/R are thought to involve a complex interaction of immediate cellular damage caused by different mediators.

In the process of prolonged ischemia, a major caused of primary liver dysfunction in donor grafts results from the generation of reactive oxygen species (ROS) which are associated to pathogenesis of inflammatory disorders including I/R injury.

From the mechanism of Ozone Oxidative Preconditioning (OzoneOP) invoked by our research group, it is shown the protective effects of ozone on important enzymes as Xanthine Deshidrogenase-Xanthine Oxidase and Superoxide Dismutase: CuZn/SOD and Mn/SOD, among others).

It is demonstrated that Ozone OP regulate the Nitric Oxide levels and other mediators which take part in the cellular redox balance.

Ozone OP participation in the novo protein synthesis as protector mechanism for morphological integrity of the liver in the preconditionated groups is shown.

The protection conferred by ozone treatment indicates that this therapeutic alternative may be successful against hepatic damage mediated by I/R associated to liver transplantation.

OZ-P-130: EVALUATION OF O3 OLEOZON ® TOXICITY TEST IN CLASS.

National Center for Laboratory Animal Production, Center for Toxicology and Animal Experimentation,
Ozone Research Center.

The toxicity test class is an alternative assay in vivo for classifying the substance as its range of toxicity. In this case allowed to classify the OLEOZON ® . For which 12 rats were used in the subline Cenp: SPRD randomly distributed at the rate of six animals per experimental group, three of each sex. 

The animals were housed in plastic boxes of polycarbonate (Makrolon) type Tecniplast T 4 , maintained under conditions suitable for the species, with a temperature of 20 ± 4 ° C, relative humidity 70 ± 10% and photoperiod 12 h light-12 h darkness. The dosing was performed in a single dose via oral gavage at a rate of 2 000 mg / kg body weight of animals prior fasting for 12 h. 

The control group was administered the vehicle (sunflower oil) and treated (OLEOZON ® ). Were recorded daily clinical signs and body weight of individual animals prior to administration, and at 7 and 14 days of the trial. 

This substance caused no deaths or clinical signs, there was no difference in body weight, showed no gross lesions in cavities or organs at this dose. In sane with the Classification System of Acute Toxicity, according to the classification system, the substance is not classified, the DL50 is> 2 000 mg / kg body weight.

OZ-P-064: DNA DAMAGE IN PERIPHERAL BLOOD LEUKOCYTES AND exfoliated cell COLORECTAL MUCOSA OF RATS TREATED WITH O3 OZONE.

Centre for Biologics Evaluation and Research, Institute of Food and Drugs,
Biomedical Research Centre.

Treatment with ozone (O3) has gained prominence because of its efficacy in various diseases associated with deficits in antioxidant defenses. However, this has proven toxic gas inhalation and have been reported studies have demonstrated its mutagenic and genotoxic in microorganisms in animals and human cells in vitro. Colorectal insufflation of the pathways is the most widely used in therapy application and whereby the mucosal cells are directly exposed to the agent. 

To study the effect of treatment on colorectal cells as well as the effect on peripheral blood leukocytes was used the rat as an experimental model, which has been used successfully in other trials with O3. Rats were treated for 4 d with 42 mg / mL for body weight. It also used an untreated control (NT) and a positive control with bleomycin (BLM).

Samples were taken after different treatments with the gas. DNA damage occurred both in exfoliated cells and in leukocytes and showed a gradual increase of the length of migration and the percentage of damaged cells as a function of sampling time. 

The samples after 48 h of treatment indicate a trend toward recovery of the damage or removal of damaged cells nonviable. 

Through this study it is found that O3 damage generated both in cells directly exposed, as in leukocytes. In the latter case is probably due to the action of its reactive intermediates.

OZ-P-067: RECTAL ADMINISTRATION OF O3 OZONE ON INTRAPERITONEAL aseptic inflammation. 

Mary O. Fox (a), Sarai River (a), Silvia Menendez (b), Ernesto Barber (a), Zullyt Zamora (b), Siegfried Schulz (c) and Maritza Victorio (a). 

(a) Institute of Basic Medical Sciences Preclinical,
(b) Ozone Research Center,
(c) University of Marburg, Germany.

Was used aseptic intraperitoneal inflammation model in order to study the effects of ozone therapy could have on the phagocytic activity. Were taken 36 female Wistar rats of 250 ± 18 g body weight and divided into two study groups: experimental (E), 18 rats treated with ozone for 5 gives rectal a daily dose of 24 mg / kg body weight and control (C), 18 rats treated with oxygen for 5 d rectal. 

At the end of treatment, all rats were implanted them under aseptic conditions, intraperitoneally, two sheets coverslips and then closed cavity. These slides were taken at 24 h (subgroups C1 and E1), at 3 (subgroup C3 and E3) and 4 d (subgroups C4 and E4), each subgroup containing 6 animals. Two hours before removal of the plates were inoculated with 0.5 mL of washed RBCs in saline rooster, which enable the evaluation of the phagocytic activity of mononuclear cell inflammation present in the sheets. The rats were anesthetized with a mixture xilaxin / ketamine, prior to extraction of the sheets. The cells attached to these sheets were fixed and set with reagent-May Grümwald Giensa and mounted on the corresponding slide. 

Readings were taken with a light microscope in successive fields to reach 100 mononuclear cells. It also measured tumor necrosis factor (TNF) in subgroups E1, E3, E4 and a control group where only the anesthetic was applied, since it is known that drugs can have some influence on the TNF. 

Yielded a significant increase (p <0.005) in phagocytic index in the subgroups E1, E3 and E4 with respect to the subgroups C1, C3 and C4, as well as the number of cells that had phagocytosed. With regard to TNF was not observed increase thereof in either group pretreated with ozone or the group with anesthetic alone.

OZ-P-068: OPTIMIZING TREATMENT OF O3 OZONE IN THE RENAL ischemia-reperfusion.

Ozone Research Center,
Institute of Preclinical Basic Sciences,
Center for Biologics Evaluation and Research Institute of Pharmacy and Food.

There are no known methods to predict which organ will work better after having had warm and cold ischemia followed by reperfusion. It was our purpose in this paper to assess the effect of ozone on rat kidneys subjected to warm ischemia, and compare the function of kidneys previously treated with ozone compared to other groups. Another objective was constituted to assess the effect of ozone on direct and indirect indicators of oxidative stress and determine the optimal period of treatment with ozone. 

Our work took 80 Wistar rats of 250 g, female, grouped into 8 sets of 10 animals each which caused him renal ischemia for 30 min, followed by 3 h of reperfusion and flow midiéndosele renal plasma and the intensity of glomerular filtration and functional variables and biochemical variables by spectrophotometric techniques deterninó superoxide dismutase (SOD), phospholipase A2 and fructosamine.

Morphological studies were also conducted to determine the structural changes that appeared during the experiment. The working groups were distributed as follows: healthy control group (I), ischemic control group (II), oxygen control group at 5 d (III), oxygen control group at 10 d (IV), control group oxygen at 15 d (V), ozone group at 5 d (VI), ozone group at 10 d (VII), ozone group at 15 d (VIII). 

The results showed a marked decrease in renal plasma flow and glomerular filtration rate in all groups (control ischemic, control oxygen of 5, 10 and 15 d of treatment, except for the group previously treated with ozone which values remained statistically similar to healthy controls.

Biochemical variables yielded equivalent results.

Structural lesions were significantly lower in the group treated with ozone at 15 d specifically in relation to ischemic controls and oxygen. interpreting the results is dependent on action to improve the rheological properties of the movement known ozone and the effect of "preconditioning" of the same protecting the body from oxidative stress.

OZ-P-069: Evolution of multiple organ injury, in an experimental model BURNED MOUSE, TREATED WITH ALOE B, O3 OZONE AND EPIDERMAL GROWTH FACTOR.

Surgical Hospital
Ozone Research Center.

Burn disease is a familiar theme of medical interest because it allows lines of research related to the sepsis syndrome, generalized systemic inflammation syndrome, multiple organ failure, shock and trauma among others. 

The aim of this study was to evaluate the evolution of multiple organ injury in a mouse experimental model of critical burn the use of Aloe B, ozone (O3) and epidermal growth factor (EGF). 

We used a dry burn model, 96 female Balb / c, divided into 4 groups: saline as control (SF), Aloe, O3 and EGF, 20 ± 2 g, 12 weeks, combined with hair removal, anesthetized with ip ketamine (150 mg / kg), 5 min before the burn. The burn was produced with an automatic welding machine with 2 bronze plaques adapted, with an area of 6 cm 2 each in the lower thoracic paravertebral region of the animal, at a temperature of 100 ° C.

The animals were resuscitated with 1 mL of SF and treated ip immediately after burn, with 15 treatments of Aloe B injection (0.3 mg/20 g in the first week and 0.2 mg/20 g in the second week , brought up to 1 mL). Were used 0.9 mL of ozone at a concentration of 37 mg / L, for 15 treatments, rectally. EGF (500 mg in 1 mL) ip, single dose. 

The SF group, 1 mL ip. In groups SF, Aloe, O 3 and EGF died 12, 13, 9 and 4 animals, respectively. Regarding vitality, in the SF group 16 animals were dying and 3 with low vitality. 

In the Aloe, 5 and 14 less active dying, but with vital signs. In the EGF group and O3 were 20 and 15 active animals dying and 4 and 7, respectively. 

In the histological study, EGF and O3 groups showed increased activity of lymphoid tissue in the spleen and minor liver and kidney damage, while the control group showed higher expression of multiple organ failure.