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1 INTERNATIONAL JOURNAL OF OZONE THERAPY formerly RIVISTA ITALIANA DI OSSIGENO-OZONOTERAPIA The Official Journal of WFOOT - World Federation of Oxygen-Ozone Therapy, FIO - Italian Federation of Ozone Therapy, SEOT - Spanish Association of Ozone Therapy, Hellenic, Indian, Slovach and Chinese National Societies VOLUME 12 - No. 1 - April 2013 CENTAURO S.r.l., BOLOGNA ISSN

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3 INTERNATIONAL JOURNAL OF Ozone THERAPY formerly RIVISTA ITALIANA DI OSSIGENO-OZONOTERAPIA The Official Journal of WFOT - World Federation of Oxygen-Ozone Therapy, FIO - Italian Federation of Ozone Therapy, SEOT - Spanish Association of Ozone Therapy, Hellenic, Indian, Slovach and Chinese National Societies Index Editorial Ischemic Stroke Penumbra 7 and Extracorporeal Ozone Treatment G. Wasser Original Articles Ozone therapy in the 16 treatment of some strictly neurologic pathologies R.M. Maffei, L.M. Maffei MR Assessment of Lumbar 25 Disk Herniation Treated with Oxygen-Ozone Diskolysis: the Role of DWI and Related ADC versus Intervertebral Disk Volumetric Analysis for Detecting Treatment Response A. Splendiani, M. Perri, A. Conchiglia, F. Fasano, G. Di Egidio, C. Masciocchi, M. Gallucci Letter to the Editor ISCO3 Data Base, a New Tool 34 for Doctors and Researchers in the Area of Ozone Therapy G. Martínez Sánchez VI Congresso Nazionale FIO Presentazione 36 G. Pellicanò Ozono e piante: 37 una difficile convivenza F. Loreto Efficacia dell ossigeno-ozono 38 terapia come trattamento di sostegno nella cura di pazienti oncologici sottoposti a chemio/ radioterapia F. Berni, A. Bolletin, A. Bonfatti, L. Chiamenti, V. De Belardini, T. De Bonis, F. Facincani, I. Kavka, G. Liaci, R. Pillai, C. Govoni L Analgesia con protossido 40 d azoto e ossigeno nella pratica ambulatoriale dell ozonoterapia per pazienti fobici, ansiosi e con bassa soglia del dolore T. De Bonis, F. Berni, A. Bollettin, A. Bonfatti, G. Chiamenti, V. de Belardini, F. Donati, F. Facincani, I. Kavka, G. Liaci, R. Pillai Plasma autologo ricco 42 di piastrine attivato con ozono: studio preliminare sulla ricrescita dei capelli The effect of ozone activated autologous platelet-rich plasma injection on pattern hair loss: a preliminary study G. Martínez-Sánchez, V. Ricci Cover: Lightning in the city, by Matteo Bonetti Indexed in: EMBASE, Elsevier (http://www.scopus.com), Google Scholar (http://scholar.google.com)

4 Effetti dell ozono terapia 43 sugli indici omeostatici e dello stress ossidativo nella malattia arteriosa coronarica (CAD) Effects of Ozone Therapy on Homeostatic and Oxidative Stress Index in Coronary Artery Disease G. Martínez-Sánchez, L. Delgado-Roche, A. Díaz-Batista, G. Pérez-Davison, L. Re Grande Autoemoterapia 44 Ozonizzata nelle ulcere vascolari croniche A. De Monte, C. Gori Ossigeno-ozonoterapia, 46 radiofrequenza e soluzioni aminoacidiche: Trattamento combinato nel ringiovanimento naturale del viso F. Berni, A. Bollettin, A. Bonfatti, L. Chiamenti, V. De Belardini, T. De Bonis, F. Facincani, I. Kavka, G. Liaci, R. Pillai Olio d oliva ozonizzato: 47 valutazione dell effetto anti-aging in vitro e della capacità ripartiva del foto danno UVB indotto in vivo M. Martinelli, G. Benevento, I. Matich, B. Bucci Ruolo dell idroozonoterapia 48 nel trattamento del piede diabetico M. Martinelli, S. Rotunno, A. Armiento, C. Valente, M. Cassol L ernia discale cervicale: 49 ozonoterapia a confronto con altre metodiche terapeutiche. Report di 736 casi V. De Belardini, F. Berni, A. Bolletin, A. Bonfatti, L. Chiamenti, T. De Bonis, F. Donati, F. Facincani, I. Kavka, G. Liaci, R. Pillai Effectiveness and Safety 51 of Oxygen-ozone Treatments for Erniated Disc: the Literature Meta-analysis Efficacia e sicurezza dei trattamenti con ossigeno ozono delle ernie discali lombari: le Meta-analisi della letteratura G. Pellicanò Open-Label, Randomized, 52 Controlled Pilot Study of the Effects of a Novel Hydrogel (Hymovis) in the Treatment of Degenerative Disc Disease F. Boraso, F. Parodi, N. Giordan Studio di sorveglianza 53 sull incidenza degli effetti collaterali indesiderati in corso di terapia infiltrativa paravertebrale con ossigeno-ozono M. Martinelli, M. Peresson, A. Armiento, S. Rotunno, M. Cassol Efficacia delle infiltrazioni 54 di Ossigeno-Ozono nel trattamento della sindrome di De Quervain nello sportivo Effectiveness of Oxygen-Ozone Injections in the Treatment of De Quervain s Syndrome in Athletes M. Moretti Efficacia delle infiltrazioni 55 con Ossigeno-ozono nel trattamento dell epicondilite nello sportivo Effectiveness of Ozone And Hyaluronic Acid in the Epicondylitis in Athletes M. Moretti La versatilità della terapia 60 con ozono nelle patologie di spalla V. Covi, G. Tabaracci Trattamento combinato 61 con ossigeno-ozono eco-guidato della patologia calcifica della spalla C. Morosi, M. Marras Iniezione intra-articolare 63 di hylan g-f 20 (synvisc ) ed ossigeno-ozono sotto guida ecografica nell artrosi dell anca, valutazione dell efficacia in 100 casi dal 2006 al 2012 C. Pasquali, C. Montoli, A. Zambello

5 L approccio conservativo 65 al trattamento dell osteoartrosi d anca. Ruolo della terapia manuale e dell esercizio terapeutico A. Romeo A, S. Borghi, C. Vanti La gonartrosi come malattia 66 sociale. La terapia con ozono come possibile soluzione G. Tabaracci, V. Covi Impiego di acido ialuronico 72 HA intra articolare in ossigeno ozonoterapia (O 2 O 3 ) per il trattamento di patologie articolari e tendinee nel paziente artrosico, novità e prospettive future integrate R. Cardelli, M. Micucci Nuovo metodo per l ossigeno 74 ozonoterapia nelle infezioni vaginali e vescicali: risultati preliminari F. Donati Herniated Discs Unchanged 83 Over the Time: Size Reduced after Oxygen-Ozone Therapy M. Bonetti Epidemiology, Economics 86 and Psycho-social of Low Back Pain B. Costa Gomes, R. Izzo, F. Zeccolini, M. Muto Organizzazione 90 di un ambulatorio per Ossigeno-ozonoterapia M. Bonetti 2013 Francesco Riccardo Monti 95 Prize Information & Congresses 6 WFOT Application Form 104 Subscription Form 105 FIO Application Form 106 Instructions for Authors 108

6 LA FORZA DELLA NATURA, LA SALUTE DELL ORGANISMO. Medozon compact Per ozonoterapia e autoemoterapia normobarica Il nuovo Medozon compact è un generatore di ozono prodotto dalla Herrmann, azienda tedesca che vanta un'esperienza trentennale nel settore, secondo gli standard tecnologici più avanzati. Si caratterizza per le elevate prestazioni, l'ottimo rapporto qualità/prezzo, le dimensioni sufficientemente compatte e un design innovativo. La presenza del touchscreen consente un'estrema semplicità di utilizzo e guida il medico passo dopo passo nell'impostazione delle percentuali di formazione dell'ozono. Integrabile con Aquazon, accessorio che consente un rapido arricchimento dell'acqua con ozono. Concentrazione di ozono 2-80 µg/ml (predefinita con fotometro) Velocità flusso 0,8-1 l/min Dimensioni (l x a x p) 424 x 185 x 388 mm Peso 13,1 kg Categoria prodotto II a CE 0123 (direttive MP 93/42 EWG) T named.it

7 meditalia srl - tel MEDITAIA srl - tel Kit ozonoterapia Solutran 0 3 ComuniCazione: La Società Meditalia, produttrice di materiale plastico, ha sentito l esigenza come altre società, di preparare un kit per Autoemo in Ozonoterapia con materiale privo di ftalati. È stato quindi superato l ostacolo della Circolare Ministeriale 1999 che proibiva l uso di sacche standard nell Autoemo perché non testate per quanto riguardava il rilascio di ftalati. Oggi gli Ozonoterapisti devono porre la loro attenzione sul confronto plastica-vetro. Per fare questo è necessario partire da una realtà: nella prima parte l Autoemo è da considerarsi prelievo di sangue intero da paziente e come tale è preferibile seguire le Disposizioni Ministeriali che regolano la trasfusione. Perché sacche -si: 1) Registrate specificatamente per Autoemo. 2) Prelievo a caduta fisiologico. 3) Prelievo reimpostato in ml. 4) Prelievo continuamente basculato. Questo implica l utilizzo di una piccola bilancia automatizzata Perché flacone-no: 1) Prelievo forzato (vuoto d aria interno). 2) Possibilità di collabire la vena del paziente. 3) Emolisi significativa dei globuli rossi. Il sangue intero infatti entrando velocemente sbatte violentemente contro le pareti rigide del flacone. 4) Procedimento che si allontana dal circuito chiuso. Cito due decreti ministeriali, tra i tanti, emanati e pubblicati nella Gazzeta Ufficiale: 1) prende in considerazione il materiale da usare - sacche con inserito anticoagulante. 2) al punto T4 indica la procedura da seguire per il prelievo. Per quanto detto e citato il flacone è stato abbandonato già dagli anni 70 nel settore trasfusionale sia a livello nazionale che internazionale. A voi Gentili Medici Ozonoterapisti riflettere sulla prima parte dell Autoemo, il prelievo di sangue intero. Cordiali saluti Fabio Malipiero Tecnico di pratiche emoinfusionali Tel

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9 International Journal of Ozone Therapy 12: 7-15, 2013 Editorial Ischemic Stroke Penumbra and Extracorporeal Ozone Treatment G. Wasser Internal Medicine Department, Chinese Federation of Ozone Therapy; Neukirchen-Vluyn, Germany Key words: ischemic stroke, penumbra, ozone therapy SUMMARY The course of events in ischemic strokes is normally seen from a point in which the penumbra is already in place. Since there is no known treatment for edema reduction, mainstream medicine focuses on re-opening the occluded vessel. Here we show that reducing the penumbra saves neuronal units from undergoing apoptosis. Introduction The treatment and treatment outcome of ischemic brain strokes is still unsatisfactory. Even medical associations dismiss major progress in the current means of counteracting the disease 1. The number of totally disabled patients after ischemic stroke has remained high over the decades at about 17% and without any sign of alleviation. Another point is that mainstream medicine does not focus on the first event after stroke onset because the first stage of inflammation, the incoming penumbra or perifocal edema, is not identified as a target for intervention since no medication exists. Physiology and pathophysiology of the neuronal unit The neuronal unit is composed of neurons, astrocytes, and specialized endothelial cells and pericytes building the blood brain barrier (BBB). The microglial cells patrolling the tissue as guards belong to the wider environment. It is common knowledge that the brain metabolizes mainly glucose and it is taken for granted that neurons are a part of glucose users. In 1999 Magistretti et al. reported that neurons mainly metabolize lactate and are widely devoid of the glycolytic pathways 2. In 2004 another publication by Pellerin and Magistretti investigated the topic further and proved the till then theoretical explanation 3. The reason is that neurons are the racehorses of our brain needing high amounts of energy to do their work in firing and transmission of excitations. The exocytic release of neurotransmitters and their reuptake are highly energy-consuming processes. While astrocytes depend on glycolysis to create pyruvate and metabolize pyruvate via acetyl-coa in the mitochondrial tricarbon cycle, neurons use lactate (lactate contains only one H-atom more). In glycolysis, phosphofructokinase-1, the enzyme creating fructose-1,6-diphosphate, is the limiting factor. This enzyme is the bottleneck in glycolysis. If neurons depended on the glycolysis to create ATP they could not work as high-speed transmitters and would be disabled in their firing capability. Nature found the solution in providing neurons with lactate enabling them to feed their mitochondria located close to the neuronal spines with fast food, lactate. Even other glial cells, like microglial cells, react promptly in case of lesions in the neuronal tissue. After laser occlusion of a single capillary the microglial cells occupy the area in a time as short as seven minutes 4. Ischemic stroke penumbra From the efficiency point of view, this division of metabolism is very effective under physiological conditions. But it creates an Achilles heel in case Reprinted with permission from The Neuroradiology Journal 26,

10 Editorial: Ischemic Stroke Penumbra and Extracorporeal Ozone Treatment G. Wasser A B C D Figure 1 A) Intact arteria cerebri media. B) Ischemic stroke and area of first impact. C) Widening of depolarized cells by potassium efflux. D) Penumbra. of disturbance and inflammation. Any obstruction in oxygen supply leads to ATP depletion and within eight minutes to necrosis. Necrosis is induced in neurons by ATP deficiency close to zero, resulting in instant cell death 5. This is why we find at least a scar in the location of the first impact of ischemic stroke in most cases. Neuronal death sends a wave of potassium into the neighborhood depolarizing the cells and creating the penumbra. In the penumbra zone cells can survive since the BBB is still intact and astrocytes and glial cells still have the means to create ATP by disabled but still functioning electron transport chains (ETCs). The relative resistance to ischemia is based on glycogen storage and use. In this area the cells will undergo apoptosis. This means that ETC complex IV still works providing the mitochondria with minimal ATP amounts enabling the cells to undergo controlled destruction, apoptosis 6. The mitochondria still maintain a mitochondrial membrane potential (mδψ) as part of their limited survival ability. Within 24 to 48 hours the penumbra passes the stages of apoptosis. But within this time with the special means of ATP delivery we can revitalize the area (Figure 1A-D). The ATP content (Figure 2) in the MCA area is close to zero after occlusion and never reaches the former level after reperfusion. The content does not equal zero since the astrocytes still produce ATP by their ETC. Here, in the middle cerebral 8

11 International Journal of Ozone Therapy 12: 7-15, 2013 Figure 2 Yellow bars show ATP deprivation after first impact, recovery of ATP content depends on astrocytes using glycolysis. Green bars show ATP in the penumbra area with ATP production continuing via ETC complex IV in astrocytes. Figure 3 Yellow bars show excessive lactate production after 60 min ischemia in the necrotic zone of neurons with still surviving astrocytes. Green bars show minor changes in the penumbra regarding lactate. 9

12 Editorial: Ischemic Stroke Penumbra and Extracorporeal Ozone Treatment G. Wasser Figure 4 Focusing on blue bars (male), 40% of the total body water is located intracellularly. artery (MCA) area the astrocytes will also develop apoptosis, since the effector cells, the neurons passed away. In the penumbra area we still find higher amounts of ATP for the same reason. If we look at the lactate content in the MCA area and penumbra we will find a vital difference (Figure 3). The MCA area produced in an animal trial by occlusion of the MCA shows an overload of lactate 5. This phenomenon mirrors the fact of surviving astrocytes with glycolysis and ETC partially functioning and the surplus of lactate results in the lack of surviving neurons which normally metabolizes the substrate. A larger portion of lactate is derived from the fact that astrocytes themselves are no longer able to use pyruvate because of cell membrane depolarization despite the fact that there is no lack of oxygen in the penumbra area. Penumbra water Researchers and clinicians consider penumbra edema a major problem. Still some clinicians infuse hyperbaric glucose or mannitol solutions to get rid of the water. If we look at the distribution of body water (Figure 4) we find that in males 40% of total body water is located intracellularly 7. From here we can assume that the edema water mainly comes from the depolarized cells and not out of the vasa. We have good reason to assume so, since the BBB is widely intact, most of the water is stored (40%) inside cells, only 16% reside in the intercellular space, and edema occurs even after total occlusion of the MCA. So the conclusion is: in the first period after ischemia the edema water comes from the affected cells. Later, after the end of the apoptotic pathways, the BBB will break down and then the edema becomes vasogenic. This view is shared by publications telling us that When vascular occlusion occurs, edema initially results in part from shifts in ions between the intracellular and extracellular compartments; this change leads to an accumulation of excess water within astrocytes and other cells, a change that in turn leads to swelling 8. On the other hand, we know by the function of aquaporins that these channels transport water without allowing protons to switch membrane side, here in the form of H 3 O +, and to pass their channel. In this way aquaporins do not disturb the membrane polarisation. The unsolved question is whether they are held in check by phosphorylation like the ATPdependent K + channel. In case of depolarization they would shed the water into the interstitium 10

13 International Journal of Ozone Therapy 12: 7-15, 2013 Figure 5 Successive steps in ATP production. See text for explanation. producing the penumbra. Otherwise the different water distribution in concert with the osmotic pressure of proteins would alter the normal arrangement 9,10. By removing the edema water early after ischemic stroke physicians close the door to revitalizing the stunned astrocytes, glial cells, and neurons. Timetable of ischemic stroke Back to the beginning of ischemic stroke: in most cases only a small artery or some capillaries are involved. Occlusion of supplying arteries like the MCA leads to death. The penumbra emerges as described above and is still perfused by mainly intact capillaries of the BBB. Only the small spot of first impact does not show any blood flow. The penumbra expands until it comes to the point that the cells are able to supply themselves with enough ATP to run their ion channels to maintain their membrane potential. This struggle creates the outer borderline of the penumbra. In the penumbra area the BBB is still intact during the time of apoptosis and glucose and oxygen is delivered but not taken up by the cell due to depolarization of the cell membranes and the stunned metabolism. Only the small obstructed vessel shows no perfusion signs. To restart the metabolism of the stunned cells ATP is needed. Glycolysis starts with the input of two mol ATP (the first to create glucose-6-phosphate and the second to create fructose-1.6-diphosphate), and exactly these two mol of ATP per mol glucose are missing in the penumbra to kick start metabolism (Figure 5). Conventional mainstream medical treatment Let us now look at conventional treatment for brain strokes. Guidelines limit the use of rtpa to four hours after onset of brain stroke. Approximately 7% of the victims reach a clinic within this therapeutic window. Even if the rtpa application reopens the occluded vessel the situation is unchanged. The latest publication records a clinical trial with rtpa and rtpa in combination with endovascular therapy 11. The first trial was stopped because there was no advantage of the combined therapy. Another paper published in the same edition of NEJM demonstrated no advantage of endovascular therapy over rtpa treatment 12. Depolarized cells in the penumbra are supplied by glucose and oxygen even without treatment or ambitions to reopen the single 11

14 Editorial: Ischemic Stroke Penumbra and Extracorporeal Ozone Treatment G. Wasser A B C D Figure 6 A) The penumbra is still perfused as the BBB is still intact. B) The extent of scarring in conventional treatment. C) Shrinking penumbra after ATP delivery. D) Spot of scarring, identical to the first impact and necrosis after ozone therapy. occluded vessel. But in the wake of depolarization they cannot start using the supply. Inflammation has started in the zone behind the occlusion and in the penumbra. The cellular environment becomes increasingly acidic with activation of metalloproteases which in turn will use the newly incoming oxygen supply to create radicals after successful re-opening of the vessel. These radicals extend the till then limited tissue damage (undergoing apoptosis) into a larger area of necrotic debris or inflict bleeding resulting in a much more extended scarring. This phenomenon is well-documented and known as reperfusion injury. The therapeutic window of four hours tries to limit the damage caused by reperfusion injury. But this treatment does not restart the metabolism in the penumbra. Warnings have been published by the American Society of Intensive Medicine not to use this method at all 1. Only a small percentage of patients treated with rtpa because of the therapeutic window seem to justify this treatment. And the evidence of induction of bleeding should be kept in mind. The end is that in most conventionally treated cases the area of first impact and the penumbra undergo scarring leading to extended loss of function and disability. Basis for explanation of ozone therapy Ozone is simply a small molecule which has only one intention: the oxidation of organic chemical double bonds. Ozone gas cannot be produced 12

15 International Journal of Ozone Therapy 12: 7-15, 2013 enriched to a hundred percent with ozone. It always contains oxygen, normally in the range of up to 97%. Therefore the gas should not be directly injected into the veins or arteries since it will disappear in ms encountering the first cell membrane. Instead we use major auto-hemo-therapy (MAHT) to induce intracellular ATP stocks and use mainly the RBCs as transporters for ATP. RBCs supply and maintain the serum with a threshold of 1 μmol/ml blood signal undisturbed metabolism 13. This ability of RBCs to disperse ATP at the area of need is used for therapy. Here the unsaturated fatty acids mostly oleic acids of the membrane phospholipids will react in ms with the ozone molecule to build ozonides. The presence of protecting enzymes inside and outside the cell dismantles the ozonides (= peroxides) into compounds no longer threatening to damage cellular structures. One of the main enzymes is glutathione peroxidase-reductase, a member of the glutathione peroxidase family, containing a selenium molecule inside the active center. The glutathione peroxidase-reductase defuses ozonides with glutathione as the substrate and proton donor into water, alcohols, and other compounds. In this reaction the reduced glutathione is transformed into the oxidized form. Since there are only limited amounts of glutathione per cell available most of the compound is found in RBCs 7 the oxidized form must be regenerated to gain reduced glutathione to start or to be ready for the next round of detoxification. By means of NADPH as proton donator, reduced glutathione is recycled. NADP +, the leftover of this reaction, changes the redox potential of the cell and ignites another round of regeneration. NADP + is transformed into NADPH in the first two steps of the pentose shunt. The limiting enzyme is glucose-6-phosphate-dehydrogenase (G-6-PD) which starts the pentose shunt. A lack of this enzyme more frequently found in Asia than in Europe disables the detoxification of radicals by glutathione peroxidase-reductase leading to hemolysis in the bottle or blood bag in case ozone gas is used. Of course there is no absolute G-6-PD deficiency since this would have devastating effects on all cellular functions. RBCs control Yin and Yang RBCs filled with ATP travel through the capillaries of the BBB sensing the low ATP content (< 1 μmol/ml full blood) and start giving away their ATP 13 which is instantly used by the stunned cells. All of a sudden there is energy available to create glucose-6-phosphate and fructose-1,6-diphosphate to restart glycolysis. With the first pyruvate available and the ion channels running in the cell membrane, re-polarisation of the double layer occurs, and ATP is generated by the ETC which did not lack oxygen but pyruvate or acetyl-coa in the penumbra area. ATP and effect on penumbra The earlier the penumbra is resupplied with ATP, the more visible is the instant effect on the clinical picture. In the first hours after stroke onset one can see the paralysis resolving. And the effect lasts with no blowback. The edema shrinks and only the area of first impact, the area of necrosis and debris from decaying neurons will be visible in MRI as a minor scar (Figure 6D). Outcome in a pilot study About 15 years ago I published 45 cases of ischemic brain strokes treated with extracorporeal administered ozone gas to full blood 14. The outcome was judged by three stages: 1) full recovery, 2) disabled with loss of fine motor function, but able to control daily life, 3) fully disabled (Figure 7). The Framingham Study shows over decades an outcome of 17% of the patients in stage 3 with no signs of decline even after the introduction of rtpa therapy. In the pilot study there was no patient enlisted in the stage 3 group. I am well aware that we cannot compare 45 patients with the high numbers of the Framingham Study, but if we can reduce the outcome in stage 3 by 5-10%, this would be a major step forward for the stroke victims to live a decent life after ischemic stroke. Therapeutic methodology with MAHT Ozone concentration and frequency of treatment: since ozone is not a medication with a dosage/efficacy ratio we must try to produce as much ATP inside cells, here RBCs, as possible to influence the perifocal edema via ATP delivery by RBCs. According to the SFDA in Beijing the highest concentration of ozone in oxygen gas is limited to 60 μmg/ml to prevent the overproduction of lysophospholipids which would interrupt membrane integrity. The effect would be hemolysis of blood cells or the necrosis of other cells. The application of MAHT is recommended with 60 μmg/ ml (equivalent to 3% ozone in 97 % oxygen) in 50 ml of blood. The surplus of oxygen will stay in the 13

16 Editorial: Ischemic Stroke Penumbra and Extracorporeal Ozone Treatment G. Wasser Figure 7 Stage 1 after stroke, full recovery. Stage II, disabled but managing daily life activities. Stage III totally disabled and immobilized. supernatant and is not used for therapeutic intervention. This lack of oxygen use prevents reperfusion injury. The often used term oxygen-ozone therapy is therefore the wrong expression and does not match the biochemical reactions. In principle and with a look at the pathophysiology of the penumbra development one extracorporeal ozone treatment is sufficient. But since ischemic stroke patients are multimorbid six to ten treatments, once on the following days, are recommended. Conclusion Because of the described rationale, we believe that using the physiological potential of RBCs to supply suffering tissues with ATP we can influence the outcome after ischemic stroke. The therapeutic window is extended to up to 48 hours until irreversible damage occurs in the sense of extended scarring. This method is safe and without side- effects since we use only the glycolysis to induce higher ATP levels in RBCs. Of course a large randomized study is advisable to better evaluate this therapeutic proposal. Acknowledgements The author and Marco Leonardi, NRJ editor, duly acknowledge the great help of numerous Chinese hospitals, only to name the Liao Ning Stroke Treatment Centre, Sujiatun District, Shenyang, and ANSCHAN CHAND DA Hospital which showed us the largest number of cases treated with the techniques here described. We also thank Luo Xiaoguang, Neurology Department, First Affiliated Hospital of China Medical University, director He Zhiyi, for the initiation of a controlled, double blinded clinical trial. And our thanks go to Pei Qi, who made all the necessary contacts. 14

17 International Journal of Ozone Therapy 12: 7-15, 2013 References 1 Alexandrov AV, Schellinger PD, Saqqur M, et al. Reperfusion and outcomes in penumbra vs. systemic tissue plasminogen activator clinical trials. Int J Stroke. 2011; 6 (2): Magistretti PJ, Pellerin L, Rothman DL, et al. Energy on demand. Science. 1999; 283 (5401): Pellerin L, Magistretti PJ. Neuroscience. Let there be (NADH) light. Science. 2004; 305 (5680): Nimmerjahn A. Kirchhoff F, Helmchen F, et al. Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science. 2005; 308 (5726): Welsh FA. Regional expression of immediate-early genes and heat-shock genes after cerebral ischemia. Ann N Y Acad Sci. 1994; 723: Kushnareva Y, Newmeyer DD. Bioenergetics and cell death. Ann N Y Acad Sci. 2010; 1201: Wissenschaftliche Tabellen - Documenta Geigy. 7. Auflage. Diem K, Lentner C eds. Stuttgart: Georg Thieme Verlag; Stam J. Thrombosis of the cerebral veins and sinuses. N Engl J Med. 2005; 352 (17): Berendsen HJ. Bioinformatics. Reality simulation--observe while it happens. Science. 2001; de Groot BL, Grubmüller H. Water permeation across biological membranes: mechanism and dynamics of aquaporin-1 and GlpF. Science. 2001; 294 (5550): Broderick JP, Palesch YY, Demchuk AM, et al. Endovascular therapy after intravenous t-pa versus t-pa alone for stroke. N Engl J Med. 2013; 368 (10): Ciccone A, Valvassori L, Nichelatti M, et al. Endovascular treatment for acute ischemic stroke. N Engl J Med. 2013; 368 (10): Rapaport E, Fontaine J. Anticancer activities of adenine nucleotides in mice are mediated through expansion of erythrocyte ATP pools. Proc Natl Acad Sci U S A. 1989; 86 (5): Wasser GH. Zerebrale Durchblutungsstörungen. In: Ozon- Handbuch. Grundlagen. Prävention. Therapie. Beck EG, Viebahn-Hänsler R, eds. Landsberg: Ecomed; p. V V Gerhard Wasser, MD Internal Medicine Department Chinese Federation of Ozone Therapy Franz-Marc-Strasse, 12 Neukirchen-Vluyn Germany 15

18 International Journal of Ozone Therapy 12: 16-24, Ozone Therapy in the Treatment of Some Strictly Neurologic Pathologies R.M. Maffei, L.M. Maffei Servizio di Neurologia, Casa di cura Villa dei Fiori ; Naples, Italy Key words: oxygen-ozone therapy, neurological pathology SUMMARY - In this work the Authors analyzed the benefits and the results of the Ozonate Autoemotherapy through two procedures: Major Auto-Emotherapy (MAHT) and Platelet Rich Plasma (PRP) in patients affected by some strictly neurologic pathologies. The patients observed, even if they were affected by very different pathologies (Parkinson Disease with the interest of Nucleus of the Base and Spine- Cerebellum Atassy with prevalent interest in Spinal Cord and Cerebellum), had in common an altered Redox Balance because of an evident Stress Oxidative verified by scientific test. In this work the Authors analyzed, therefore the action of Autoemotherapy headed for contrasting the so called Free Radicals, which are the products and the true responsible of the Oxidation and that marked the different mechanism of the two components of Autoemotherapy: MAHT and PRP. Introduction It is now generally accepted that the Oxidative Action (that takes place in our body and is important to keep some vital processes when included in physiological parameters), when excessive, can be significantly detrimental to determine pathological processes including some special severe condition as the Oxidative Stress. We have to clarify that, nowadays, does not exist a real pharmacologic therapy that can effectively fight oxidative stress, except for the one that use the natural antioxidants ingested with foods like certain vitamins (E, C, etc..) which, however, have not yet shown any real effects. Against this deficiency a valuable help comes from autohemotherapy in both the two forms: MAHT (Major Auto-Hemo-Therapy) and PRP (Platelet Rich Plasma). The more realistic and fascinating Hypothesis about the effectiveness of MAHT against the harmful effects of a high oxidative stress is certainly the one developed by Prof. Velio Bocci. Simplifying the theory of prof. Bocci we can say that it is rooted in the observation that if free radicals (oxidative stress real responsible) exceed a certain alert level in a slow and gradual way, they are not perceived by the endogenous antioxidant system as "aggressive", so the body does not react to them, as it considers the condition created still within physiological limits, on the contrary to what suddenly happens for the same abrupt increase in acute or traumatic events of any kind insurgents. A striking example of the above statement is offered by the latest theory on the genesis of Parkinson s Disease which is based precisely on the fact that the onset of a chronic oxidative stress at the level of the Substantia Nigra of Sommering would lead to the destruction of mitochondria of cells nigral with subsequent cell death and reduced production of dopamine. According to prof. Bocci the mechanism of MAHT is therefore based on the fact that, at least in chronic diseases and in deseases where there is no genetic damage that affects specifically the endogenous antioxidant system (See below a short explanatory note) a sudden increase of free radicals even if slight and subliminal, occurring with the entry into the circulation in a short span of time of blood ozonated around g/ ml would determine a reaction by the endogenous antioxidant system. Consequentially there would be an increase of the production of various enzymes with antioxidant action involved in the process. This mechanism would explain the long latency between the first treatment with MAHT and its beneficial effects that would not be visible before than days. At this point, however, attentions should also be paid when the etiopathogenesis of the disease is known. 16

19 R.M. Maffei ozone Therapy in the Treatment of Some Strictly Neurologic Pathologies An example is provided by the Amyotrophic Lateral Sclerosis. In this disease, in fact, the MAHT might be contraindicated if not even considered detrimental in those forms in which it has been hypothesized, like pathogenetic mechanism, a genetic deficiency of the production or the enzyme superoxide dismutase (Cr 21) or, according to peroxidative hypotheses, a reduction of glutathione reductase activity. Special attention deserves the Amyotrophic Lateral Sclerosis (ALS) where, precisely in at least one form of it, one of the most reliable and recent pathogenetic hypotheses would trace the primary cause of the disease in question in a lack of probable genetic nature, that is the production of glutathione reductase (an enzyme known to play a major role in controlling the redox balance as very important element in the so-called endogenous antioxidant system) and where, therefore, the MAT is ineffective or even contraindicated. According to the authors of this work the hypothesis that supports the use of the PRP against Oxidative Stress lies because some essential and fundamental components of the blood (erythrocytes and platelets) contain a rich set of endogenous antioxidants, especially some catalase which, activated and suddenly inoculated after a simple injection intramuscularly, exert a direct action even more rapid against oxidative stress with a duration in time significantly lower than the MAHT. In particular it is necessary here to recall that, as universally known, all the cells of an organism are capable of producing antioxidants enzymes and such action will occur each time a cell produces free radicals in excess. In such conditions, with a mechanism that could be called on-demand, when free radicals exceed a certain threshold limit, there would be a response by the cell affected by the process characterized by a specific and targeted production of antioxidant enzymes sufficient to neutralize the momentary aggressive oxidative action so that the body can defend from the harmful effects of free radicals present in excess in the same cell. To this rule, of course, make exception corpuscular elements of blood, such as Platelets and Erythrocytes, which haven't any nucleus and therefore are not able to produce antioxidant enzymes independently, continuously or on demand. However the corpuscular elements' progenitor cells provide them with antioxidant enzymes, produced in a larger quantity than what is needed, in order to let them survive even when already detached but still unable to produce independently antioxidant enzymes. At this point the Platelets and Erythrocytes use different antioxidant enzymes for different purposes: The Erythrocytes with their relatively long lifespan (approximately 120 days) will use the antioxidant enzymes substantially to defend the EME from very oxidative action because, as it is rich of iron, it has a strong tendency to oxidize if not able to bind the oxygen and therefore no longer able to carry out its mission. The Platelets, however, in their short life (4-5 days in all) basically and normally do not need that rich set of provided antioxidant enzymes, except for a small amount of them that will use them to defend their membrane from the attack of free radicals. Most of them, in fact, will be unusable and unused, unless they do not lose their membrane integrity and put at the disposal of the body their antioxidant enzymes together with all the other components of defense, as occurs, for example, in case of rupture of the endothelial barrier. Starting from these brief considerations, the authors intended to use this innovative technique of autohemotherapy by MAHT and PRP in a rational and planned way and so, as specialists in Neurology, they found it appropriate to do the same test on some patients who were suffering from diseases of neurological interest that, in their opinion, could provide an altered redox balance. The patients selected were in fact three men with Parkinson s disease and two subjects (one male and one female) affected by Spine-Cerebellar Atrophy both carriers of a sporadic form. All patients affected by diseases of the nervous system had an altered redox balance, in the sense that in all was observed, using the specific test Caratelli, a discrete Stress oxidation. At this point it is worth to point out that the present work, due both to the small number of cases treated, and to a lack of cross-checks, currently has no claim to scientific validity, but the authors, taking advantage from a careful observation of few clinical cases and good results obtained, consider it appropriate for the moment to make them objects of a simple scientific communication, hoping that it will arouse some interest and perhaps stimulate the desire to better investigate this fascinating field. Materials and Methods All the selected patients, once studied extensively and systematically classified clinically, after demonstrating at test Caratelli to present an altered redox balance, that is a discreet Oxidative Stress, were subjected to a single treatment protocol specifically defined as follows: 17

20 Ozone Therapy in the Treatment of Some Strictly Neurologic Pathologies r.m. Maffei Treatment Concentration Frequency Duration MAHT 25 g / ml 2 times per week From 1 st to 2 nd week MAHT 35 g / ml 2 times per week From 3 rd to 4 th week MAHT 40 g / ml 2 times per week From 5 th to 6 th week MAHT 40 g / ml 1 time a week From 7 th to 10 th week MAHT 40 g / ml 1 day 15 days From 11 th to 14 th week PRP 5 cc a 70 g / ml in m. Together with MAHT Together with MAHT Shown below only the date of two patients: a man with Parkinson s disease and a woman suffering from Sporadic Spine-Cerebellar Ataxia: Patient. N 1: B. N., born 10/08/46 Family History Ing neuropathological negate the noble History Physiological Pregnancy and Childbirth The second of five siblings (brothers and sisters in abs) in full-term pregnancy by eutocia normocondotta. Psycho-Motor Development normal Suitable for lever. Married with healthy offspring. Pathological History Other Pathology Common rashes s childhood. Myocardial infarction in March 1999 In the month of June 1999 he had a surgeon of coronary angiography with implantation of two stents. In 2005 he detected diabetes mellitus, the statecontrolled oral hypoglycemic agents. Specific Disease Since August 2007, the patient started to present large tremor shook aside all left upper limb, which disappeared during the sleep, slow ideo-motor, micrograph, slightly Figés facies, bradykinesia species at left upper limb. Neurological Examination Patient awake, conscious and well oriented in three dimensions Signs of meningeal irritation: none Cranial Nerves: free Tone: fairly increased waxy type Trophism and Motility: the 4 limbs preserved ROT: normal and symmetrical elicited in all four limbs Abdominal reflexes: present Babinski: absent bilaterally cerebellar tests: performed correctly Romberg: negative Gait: normal ambulant with reduced pendular movements of the upper limbs species left Psychological Examination Normal Tests Performed: MMSE: Score: 30/30 Q-EEG-Brain Mapping: The Spectral Analysis of Spontaneous Brain Electrical Activity argues for moderate aspects of cortical atrophy in fronto-parietal paramedian left. Redox Balance: Fort: 361 U (see n.: ) Ford: 1.52 mmll / l (see n.: ) Drug Therapy in Act Concor cp: cp ½ in the morning Ticlopidine cp: 1 tablet after breakfast and 1 tablet after dinner Stalevo 100: 1 hour 7 +1 cp cp 20 hours patch Neupro 4 mg: 1 patch every 24 hours Sirius cp 100 cp: 1 tablet in the morning Novoform cp: 1 tablet with meals Raliprost cp: 1 tablet per day Following are presented the rating scales for Parkinson s UPDRS at different times of life of the patient and in particular those carried out before and after treatment with ozone. Discussion on First Case From these above mentioned data it s easy to observe the wavering course of the disease, that generally responded well to drug therapy with Levodopa. 18

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