1. HOW MUCH DOES OZONE THERAPY NEED A REAL BIOCHEMICAL CONTROL SYSTEM? THE OXIDATIVE STRESS IMPORTANCE AND ASSESMENT FRANK HERNÁNDEZ Ozone Research Center. Cuba

2. Why ozone in medicine has not been approved as a common practice by world medical community? The use of specific parameters to control the applicable dose in terms of safety and therapeutic effects has been gaining importance Ozone Therapy

3. Ozone therapy and the Oxidative stress Mechanism of action Regulator of endogenous Oxidative Stress Toxic effects Therefore, the oxidative stress phenomenon is deeply involved in ozone therapy effects. A test or system for assessing oxidative stress process will be of great relevance

4. Ozone therapy and the Oxidative stress Oxidative stress can be defined as the prevalence withing the living cell of oxidizing activities over the cellular antioxidant defense activities. Complex Biological Process Different Oxidant substances Different Antioxidant compounds

5. ESPECIES REACTIVAS DEL OXIGENO Especie MolecularVida media (seg. 37 0 C HO · 1 x 10 -9 RO · 1 x 10 -6 ROO · 0.1 – 7 x 10 -3 O 2 ·- 1 x 10 -6 (SOD) 1O21O2 1 x 10 -6 NO · 0.1 - 5 H2O2H2O2 2.5 (plasma) HOCLRegularmente estable O=NOO · 1 – 4

6. Oxidative Stress from Ozone Therapy Oxidant Substances Criege Ozonides Hydrogen Peroxides Hydroxyperoxides Aldhydes Antioxidant Compounds GPx GSH GST SOD CAT ADH

8. System for Oxidative Stress Diagnosis 3 mL of Blood PlasmaErythrocytes Conjugated Dienes Thyl-Barbituric Acid Reactive Substances Total Hydroperoxides Reduced Glutathione Glutathione Peroxidase Glutathione S-Transferase Superoxide Dismutase Catalase

9. SISTEMA DE DIAGNÓSTICO DEL ESTRÉS OXIDATIVO System for Oxidative Stress diagnosis Total Antioxidant Activity Total Pro-oxidant Activity Redox Index Grade

10. PARAMETERSREFERENCE Total Antioxidant Activity (Units) 134.2 – 170.1 Actividad Prooxidante Total (Unidades) 145.0 – 169.9 Indice Redox0.924 – 1.320 Grado0 Oxidative Stress Diagnosis

11. Grades of oxidative stress and their clinical meaning Grade Meaning 0 No oxidative stress 1 Light oxidative stress 2 Moderate oxidative stress 3 Severe oxidative stress 4 Very severe oxidative stress

12. Oxidative Stress Diagnosis of cardiopathy Patient O3T Sessions Total Antioxidant Activity Total Pro- oxidant Activity Redox Index Grade 0128.38167.960.7642 5123.45178.690.6913 10138.92175.210.7902 14144.28169.700.8501

13. Using Oxidative Stress Diagnosis in Ozone therapy Grade 0: Any range of concentration and dosage. Grade 1: From top level of medium range to downward. Grade 2: From minimal level of medium range to downward. Grade 3: Minimal level of low range. Grade 4: No ozone therapy.