international journal of the official journal of wfoot ... · ) molecule has an extremely insta-ble...

formerly RIVISTA ITALIANA DI OSSIGENO-OZONOTERAPIA

INTERNATIONAL JOURNAL OF

OZONE THERAPYThe 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 10 - No. 2 - OCTOBER 2011 CENTAURO S.r.l., BOLOGNA

ISSN 1972-3539

Index Majorozonated 85 autohaeMotherapy

InthetreatMentofLIMbuLcersnotrespondIngtoconventIonaLtherapy

a.deMonte,c.gori

effectsofozonetherapy 99 onoxIdatIvestress

bIoMarkersIncoronaryarterydIseasepatIents

L.delgado-roche,g.Martínez-sánchez, a.díaz-batista,L.re

oxygen-ozonetreatMent 105ofverrucas

t.gastaldi

anewInfILtratIve 109 paravertebraLapproach

forthecureofdIschernIatIonusIngo2o3

r.viglioli

LetterstotheedItor 110

RepoRtage 111 IIIworLdcongress

ofoxIgene-ozonetherapyv°congressonazIonaLef.I.o.

Information & Congresses 84,147

WFOT Application Form 150

Subscription Form 151

FIO Application Form 152

Instructions to Authors 154

Indexed in: eMBaSe, elsevier (http://www.scopus.com), google Scholar (http://scholar.google.com)

INTERNATIONAL JOURNAL OF OZONE THERAPYThE 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

formerly RIVISTA ITALIANA DI OSSIGENO-OZONOTERAPIA

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85

www.centauro.it International Journal of ozone therapy 10:85-98,2011

Major Ozonated Autohaemotherapy in the Treatment of Limb Ulcers not Respondingto Conventional Therapy

a.deMonte1,c.gorI2

1 anaesthesia and ICU Department, azienda ospedaliero-Universitaria; Udine, Italy2 paSS outpatients Clinic; Udine, Italy

Key words:legulcer,chroniculceration,ozonetherapy,oxidativestress,regenerativemedicine

SUMMARY -Chronic leg ulceration significantly impacts on the individual’s health status. It is also a major financial burden on the national healthcare system. the maintenance of chronic ulcer is the result of an equilibrium imbalance between local tissue demand and systemic metabolic supply. the effect of this pathological condition leads to an increase in free radical production overwhelming the scavenging capac-ity of physiological antioxidant defence mechanisms. the reduction of oxygen free radicals concentration removes one of the major causes hindering the healing of chronic ulcer. Blood ozonation is recognised to induce a strong oxidative stress which causes an oxidative preconditioning that increases the efficacy of endogenous oxygen free radicals’ scavenging properties. after a brief review of the pathophysiology of cicatrization and ozone properties, we document a significant number of patients who did not respond to conventional treatment, yet were successfully cured with major ozonated autohaemotherapy.

Introduction

chroniclegulceration(cLu)isdefinedasany wound below the knee that does not heal within a six-week period1.Itisacommonpathologyaffect-ingapproximately1%of thepopulation, increas-ing up to 3% or more when considering peopleaged65andover1-11.cLuhasamajor impactonanindividual’squalityoflife,aswellashealthcaresystemcosts.healthfinancialreportsestimatethat1% of annual healthcare budgets in europeancountriesareabsorbedbylegulcertreatments.Intheusa,cLupatientsaccountformorethansixmillionpeopleandthefinancialimpactapproach-es 2 billions euro1,4. In Italy approximately twomillionspeoplehavecLu5,6.regardlessoftheeti-ology,cLuleadstotissueinflammationproducedbyinadequatebloodsupply,whichresultsintissueanoxia, oedema, induration and extravasation ofserum,cytokinesandbloodformedelements.thefinal step of this pathophysiological cascade is astructural change in the surrounding tissue andcellulardeath1,3,10.

therapeutic management of cLu includes amultidisciplinaryapproachinvolvingmedical,para-medical,surgicalandtechnologicalcompetencies8-11.

Majorozonatedautohaemotherapy(o3-Maht)has been reported as an effective treatment

of cLu. this therapy was first introduced intothe clinical field by hans wolff in 1974. sincethen, much progress and many new discover-ies have been made. although there have beenmany contributors involved in the research, wegratefully acknowledge professor velio bocci ofsienauniversitywhoserelentlessenthusiasmandresearch has created a major awareness and hasbrought to the forefront the significant medicalapplicationofo3-Maht12,-15,35-46.

this review present the results obtained in aseriesofpatientswithchronic legulcerationwhodidnotrespondtoconventionalmedicalandsur-gical therapy, but subsequently were successfullytreatedwitho3-Maht.

Healing Process

before examining the mechanism of oxygen-ozone treatment, it isbeneficial tobrieflyrefreshthepathophysiologyofthehealingprocess.

regardlessoftheetiology,anulcerationimpliesabreachintheepitheliallayercoveringtheskinsothattheunderlyingtissueisexposedanddamaged.thephysiologicalresponsesincludetheactivationof the healing process reacting to the interactionbetween platelets, exposed collagen and other

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Major ozonated autohaemotherapy in the treatment of Limb Ulcers not Responding to Conventional therapy a. De Monte

Ozone Properties

theozone(o3)moleculehasanextremelyinsta-blestructurecomposedbythreeatomsofoxygen.Initsgaseousstateo3isheavierthano2;at20°cits half-life is around 40 minutes. structural o3decayentersthesamemetabolicpathwayofoxy-gen, its triatomic molecule breaks down into theo2 molecule and atomic oxygen according to thefollowingreaction12-15:

o3+biomolecules--->o2+o°

atomic oxygen is a very reactive atom whichimmediatelyreactswiththesurroundingenviron-ment and contributes to generate the same rosregularly produced by o2 decay during normalphysiologicalprocesses.Itisnoteworthytounder-line that 1-5% of oxygen metabolized by themitochondria,leakstheoxidativephosphorylationtoundergounivalentreductionandoriginatefreeradicals which have to be detoxified by endog-enousantioxidantsdefencemechanismstopreventros injuries24-30. contact with ozone originatesa short and moderate oxidative stress that raisestherosconcentrationandtriggersanincreaseinendogenousantioxidantsystemresponse.

repeatedcontrolledexposuretoo3givesrisetoanoxidativepreconditioningwith the final effectof upregulating the body’s antioxidant defencemechanisms6-11 aimed at maintaining the cellu-lar redox balance and increasing the scavengingefficacy of ros and free radicals31-33.as a conse-quence,becauseoftherapiddisappearanceofo3due to its short half-life, the enhanced action ofendogenousantioxidantagentsismainlytargetedtoward the ros population distributed acrosstheorganismrather than toward thevolatileandinstableo3.

other reported direct and indirect effects ofo3ono2deliveryandimprovedmicrocirculationinclude the modification of cell wall membranecharacteristics and the increase in 2-3 dpg andnitric oxide production. the direct effect of 2-3dpgincreaseisareducedaffinityofhaemoglob-in foro2 that shifts to the right theoxygen-hae-moglobin dissociation curve and makes it easierfor the haemoglobin to release bound oxygen.several authors have reported a rise in fluidityand deformability of the erythrocyte membraneafterozonetreatmentwhoseultimateeffectisanimprovementinbloodrheology12.

finally,theincrementofnitricoxidegenerationis another factor improving tissue oxygenation.nitric oxide (no) is a potent signal transmitterinvolved in nearly every phase of cellular activ-ity, including inflammation processes and host

extracellularsubstances.evenifthehealingproc-ess is a continuum that begins immediately afterinjury,conventionally it isdivided into fourover-lappingphases:inflammatory,migratory,prolifera-tiveandremodelling.

the inflammatory reaction ismainlysupportedby platelets which attract the neutrophils andmonocytes intheaffectedregion.theseactivatedcells produce chemotactic agents which amplifythe attraction of other inflammatory cells likefibroblastsandothermesenchymalcells18,19.

during the migratory phase, an increase inthe local concentration of inflammatory cells,cytokines, growth factors and other mediatorsaid in improving and creating an advantageousenvironment for tissue proliferation. fibroblastsproduceextracellularmatrixandneoangiogenesisisstimulatedbylocalhypoxiaandspecificgrowthfactors.

during the proliferative phase, the regenera-tive process requires not only growing enzymesbutalsoaproperdeliveryofo2andvitaminc,astrongantioxidantagent,andavigorousenzymat-icactivity.thecontractileactionofmyofibroblastswill contribute to restrict thewoundareaand topulltogethertheedgesofthelesion.onlyifthisphase is uninterrupted, will the final remodel-ling stage take place and the collagen structurallinks be rearranged in a normal structure16-23. Ifthe metabolic demand for damaged tissue is notproperly met, the healing cascade is interruptedattheinflammatoryphaseandthelesionbecomeschronic.

achroniclesionischaracterizedbyanincreasedconcentrationofvasoactivesubstancessuchashis-tamine, bradykinin, serotonin, paf,tnf, cellulargrowth factors, adp, derivatives of arachidonicacid (prostacyclin, thomboxane, leukotrienes),interleukin, etc. chronic inflammation is alsoresponsibleforpersistingplateletsandcoagulationcascadeactivation,vascularpermeability,oedema,capillarycompression,hypoperfusion,etc.

Local ischemia is a particularly critical factor,whichhinders thehealingprocessbecause itper-petuateshypoxia,lowph,highlacticacidconcen-trationandlipo-peroxidation.oxidantmetabolitessuchasfreeradicalsandotherreactiveo2species(ros) increase dramatically in chronic hypoxicareas and become major and essential factorsof chronic lesion maintenance16-23. In physiologi-cal conditions, ros synthesis is balanced by thebreakdown activity of endogenous antioxidantdefencemechanismswhichincludeseveralagentslike superoxide dismutase, catalase, albumin, etc.when this equilibrium is lost, the toxic effect ofros becomes clinically recognizable in differentforms,uptopermanenttissuedamage.

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ofthepatients.aftersigninganinformedconsentform patients began the o3-Maht therapy.themajority of ulcers were of multifactorial origin.theaverageulcerarearangedfrom1 to150cm2

(mean30cm2).ulcerswerereportedtohavebeenopened and exposed varying from three to 60months (median 10). deep ulcers involving themuscles were observed in nine cases. pictures ofthe lesions were taken at the beginning of treat-ment,everyfive-sixsessionsandagainattheendofo3-Mahttherapy.

theo3-Mahtwasperformedasdescribedbybocci12,13. with the patients lying on a treatmentcouch, a sterile technique was used and a largebore peripheral vein cannulated with a butterfly16g needle, 225 ml of the blood was withdrawninadedicatedozone-resistantglassvacuumbottle(b-braun)containing20mlofsodiumcitrate3,8%as anticoagulant. again with a sterile technique,

defence.nohasalsoapotentdirectvasodilatingeffect and therefore it may play a crucial role incLupathophysiology34.

Materials and Methods

thecasesof17patients(12menand5women)are described below. their mean age was 74.7yearsrangingfrom37to82.fromtheonsetofthefirst examination, these patients had had chronicleg ulcerations for several months or years andallwerepoorlyrespondingtotheirstandardcon-ventionalmedical treatments.any indications foropen surgery or percutaneous revascularisationhad been excluded in all patients.three patientswereonregularinsulintreatment.duetodramaticworseningofthelesionandaninabilitytocontrolextreme pain, amputation was considered in six

A B

C D

figure1Case 1.a)rightleg;b)Leftleg;c-d)finalresult.

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effect.thepainwasunmanageabledespiteacom-binationofmajorandminoranalgesicagents.thepatient’squalityoflifewasextremelypoorduetothecontinuouspainandtheextremelimitationofdeambulation which was only possible with thehelpofcrutches.

anepiduralcatheterwasplacedforacontinu-ousbupivacaine(25mg/day)andmorphine(3mg/day) administration. the pain became control-led, however, the lesion worsened and becameinfected.asanextrememeasure,thesurgeontookintoconsiderationabilaterallegamputation.asalastresort,o3-Mahtwasinitiated.after62treat-ments the left leg wound was completely healedwhile a total of 120 sessions of o3-MaHt wererequired fora complete recoveryof the right legaswell(figure1).

Case 2. a 62-year-old type 2 diabetic womanwasbeingadministeredxvunitsof insulin twicea day. prior to our intervention, the patient hadundergoneachemical lumbarsympatectomy,andshewaswearinganepiduralcatheterforcontinu-ousadministrationofbupivacaine(18mg/day)andmorphine (2 mg/day). for three years there hadbeena largechroniculceronher right legwhichwas regularly medicated and debrided by a der-matologist. due to the worsening of the clinicalcourse a o3-Maht treatment was initiated.theeffect of 18 treatments is reported in figure 2a.after40sessionsthewoundwashealed.

case3.a72-year-oldwomandevelopedanon-healing chronic ulcer on the medial aspect ofher left malleolar zone 12 months prior to ourintervention. due to the poor results of con-

225mlofaoxygen-ozonemixturewasintroducedthroughadedicated inlet into thebottlecontain-ingthecollectedblood.theoxygen-ozonemixtureconcentration was gradually increased in the fol-lowingfourtofivetreatmentstoreachamaximumof 50 mcg/ml of ozone, corresponding to an o2concentrationof95%.toproducetheozoneano3generator(dr.haenslerozonosanphotonik)wasused,whichsynthesizedthedesiredoxygen-ozonemixturebytreatingmedicalo2withahighvoltagedischarge.after the addition of ozone, the bloodcontainingbottlewasgentlymixedfortenminutestoallowcompletebloodozonation.finally,usingatransfusiondripthebloodwasslowlyreinfusedtothepatientovera20-30minuteperiod.

these patients were treated twice a week dur-ing the first four-five weeks, followed by weeklyadministration.

patients’currenttherapyincludingtopicaldress-ingswasuninterrupted inall cases.therefore theonly therapeutic change made in these patientswastheadministrationofo3-Maht.

Case Reports

Case 1.a57-year-oldman;non-smokerandnon-drinker.there were no other associated patholo-gies reported. two years earlier he started tosufferaprogressiveworseningofdeepulcerationson both legs. the patient was evaluated by sev-eralspecialists,includingadermatologist,internist,allergologist,plasticandvascularsurgeon.allergic,vascularandautoimmunediseasewasexcluded.avasculitisofunknownoriginwasdiagnosed.

evenacourseofcortisonedidnotproduceany

A B

figure2Case 2.a)reportstheeffectof18treatments.b)finalresult.

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painandsympatheticblocktreatment.theanalge-sicmixturewastitratedtoinfusemorphine3mg/dayandbupivacaine35mg/day.this therapywasefficaciousforpaincontrol,buttheulcersdidnotsealinspiteofroutinesurgicaltoiletteandtopicalmedication.afterfivemonthsofepiduralcatheterplacement, major ozonated auto-haemotherapywasinitiatedtwiceaweek.after20treatmentshewasnolongerreceivingo2treatment.fiveweekslater,a reductionofepiduralmorphineandcath-eter removal was scheduled. after a total of 46 o3-MaHttreatments,theulcerswerecompletelysealed,thesurroundingerythemadisappearedandnormalskincolourwasrecovered(figure4b).

Case 5.an 82-year-old man had suffered withanulcerforninemonthsandwasscheduledforanautologous skingraft forulcer repair.theplasticsurgeon initially proposed o3-Maht to explorethepossibilitytodetergethebottomofthewoundbefore plastic surgery.after 13 o3-MaHt treat-ments the lesion was completely healed withoutrequiringanysurgery(figure5a-b).

Case 6. the clinical history of this 43-year-oldpatientreportedakidneyinfectionsincetheageoftwenty.thisconditionledtorenalfailurerequiringchronichaemodialysis.heunderwent twokidneytransplants,inbothcasesexperiencinggraftfailureresultinginremovaloftheorgan.subsequentlyhefirstunderwentpercutaneousangioplastydilationofacriticalfemoralarterystenosisandthenaper-cutaneousinsertionofastent.despitethissurgicaltreatment,achronicpainfululcerappearedonhisleft big toe and heel approximately three yearspriortoourintervention.apartialtoeamputation

ventional medical treatment, the patient becamediscouraged, and was non-compliant in applyingthe recommended dressings on the wound.after22 sessions of o3-MaHt the lesion had healed(figure3).

Case 4. a 77-year-old man had been a heavysmoker for 50 years (40 cigarettes a day) with ahistory of lung emphysema and the features of apinkpuffer.hepresentedapersistentproductivecough, with abundant excretion and sputum.thepatienthadbeenreceivinghomeo2therapyforafewmonths(3litres/min).hisgaitwaslimiteddueto exertional dyspnoea and lower limb pain, andhiswalkwasassistedwithcrutches.achestx-rayrevealed cardiomegaly, despite being on diuretictreatmentsforfouryears.

the patient suffered lower limb arterial insuf-ficiency and pain-related insomnia. a painfulchronic ulcer had extended over the right lateralmalleolusforthelast18months(figure4a).theulcer showed a marked red discolouration, theedges of the ulcer were rolled and the base wascoveredbyafranklypurulentmaterialanddebris.a smaller ulcer was also located on the lateralportionofthefoot.asevereexfoliativewaterydis-chargeerythemaaffectedthesurroundingareaofthefootandankle.hewasclassifiedinstageIvoftheLeriche–fountainescale.acontrastographicarteriogram excluded surgical revascularisationindications. he underwent percutaneous angi-oplasty, and intravenous prostaglandin (80 mcgtwiceaday)was infused for15dayswithoutanysignificant result.an epidural catheter was posi-tioned in the lumbar spine and connected to anelastomericpumpwith0.5ml/hofdeliveryratefor

A B

figure3Case 3.

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from her relatives, she looked for an alternativetherapeuticapproach.ozonizedautohaemothera-pywasinitiatedandcompletehealingofulcersandpainreliefwereobtainedafteronly10 o3-MaHt treatments(figure7b).

Case 8.a70-year-oldmalecarpenterwithtype-2 diabetes and receiving xxxv units of insulintreatment daily had had a chronic small butextremely deep painful ulcer in his left heel forthe past 12 months.the ulcer appeared purplish,with rolled margins, a non-granulating base andawaterydischarge(figure8a).twiceaweekthepatient underwent major ozonated autohaemo-therapy. the pain disappeared after five treat-ments,atotalof15 o3-MaHtwasrequiredbeforetheulcerhealedcompletely.

was required due to ostenecrosis (figure 6a,c).the chronic ulceration did not heal and con-tinuouspainpersistedwithintensityafterdialysistreatment.thepatientrefusedacompletebigtoeamputation proposed by the surgeon. after twomonths no evident improvement was noted.thepatientcameformajorozoneautohaemotherapy,inwhichhereceived25 o3-MaHtallowingacom-pletehealingofulcers(figure6b,d).

Case 7. a 75-year-old woman with depressionand a sedentary lifestyle. the patient developedvenous stasis ulcers on her left leg. her surgeonadministered topical medication and curettage.however, the clinical picture worsened with theenlargement of ulcers and an increase in pain(figure 7a).after three months, under pressure

A B

figure4Case 4.

A B

figure5Case 5.

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A B

C D

figure6Case 6.

A B

figure7Case 7.

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A B

figure8Case 8.

A B

figure9Case 9.

← ←

A B

figure10Case 10.

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ments. subsequently, the planned radiotherapywas not feasible due to an incomplete healing oftheexcisedregion.thepatientcametoourobser-vationeightmonthsaftertheinitialoperation.theresults achieved were astonishing. after only 10 o3-MaHttreatments,thesurgicalwoundwasper-fectlyhealedandthepatientwasabletocontinuehistherapeuticplan(figure10).

Case 11.a 74-year-old obese man with type 2diabetes. for several months he had presenteda rapid worsening of a devastating ulcer, whichdeepenedintothemuscularplaneoftherightleg(figure 11a,c).the pain suffered by the patientwas scored as 100 on a visual analogic scaleandthepatientwasunabletowalkorstandonaweight-bearing foot. angiography excluded anycriticalvascularstenosissusceptibletosurgicalorpercutaneous intervention. ozone treatment was

Case 9.a65-year-oldwomanwithosteoarthritis.threeyearsearlier,aseveredeformityoftheleftfirst metatarsophalangeal joint required excisionand amputation of the left big toe. subsequently,in the insertionsiteof surgicalnails,anosteomy-elitis developed. the patient underwent severalantibiotictreatmentsandthenailswereremoved.however, a chronic deep ulceration remained atthe distal extremity of the first metatarsal bone.figure9(arrowindicatesacornthatdevelopedasaconsequenceofthealteredtransmissionofforceson the weight-bearing foot).all the medical andsurgicalattemptstohealtheulcerfailed.after25 o3-MaHtthewoundhadhealed.

Case 10. a 42-year-old man presented with ararecaseofMerkeltumor.thetumorwasexcisedfrom the patient’s gluteus. Insufficient healing ofthis area existed despite multiple surgical treat-

A B

C D

figure11Case 11.

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A B

C D

figure12Case 12.

A B

figure13Case 13.

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o3-Maht.figure13bdepictsthecompleteheal-ing of the wound after 20 o3-MaHt treatments.theresultsobtainedinthiscaseshouldbeempha-sized since the healing was achieved where scartissueexisted.Moreover,duringsurgeryallthesofttissueclosetothebonehadbeenexcised,limitingthephysiological capabilitiesof anormalhealingprocess. as a reaction to the healing process, astrongadhesiontotheunderlyinganatomicplaneoccurredhinderinganadequateamountof colla-gendepositionandneo-angiogenesis.

Case 14.a55-year-oldmalesufferedatraumaticfractureofhisleftankle.asurgicalfixationofthefracture was performed by a plate application.a few months later, the plate and screws wereremovedduetoanintolerance.woundhealingwasdifficultandanalternationofopeningandclosureofthewoundbecamepartofthepatient’slifeforthenexttwoyears.heunderwentseveralcoursesofantibiotictreatmentsduetoarecurrentwoundinfection.hedevelopedtheclinicalmanifestationofalgodystrophywithpain,vasomotordisturbanc-esandtrophicchanges,withamarkedswellingofthelowerleftlegonstanding.priortoo3-Maht,his wound had been constantly open during thelastninemonthsandhewasunderciproxintreat-ment because the culture of exudative effusiongrewpseudomonascolonies(figure14a).after29 o3-MaHttreatmentsweobservedclosureofthewoundasshowninfigure14b.

Case 15.a 38-year-old man. fifteen years agothe patient underwent two surgeries for arthro-desis.a few years later an ulceration formed onhisheel,whichwasinitiallytreatedconservatively

proposed as the only alternative to amputation.anepiduralcatheterwaspositionedinthelumbarspineforcontinuousadministrationofbupivacaine25 mg/day and morphine 3 mg/day. the resultsdisplayedinfigure11b,dwereobtainedafter50 o3-MaHttreatments.

Case 12. a 68-year-old man suffered a motor-bike accident which lead to the formation of anextendedhaematomainhisrightcalf.thesurgeonfollowed a conservative therapeutic protocol andthe lesion was left undrained. the haematomabecame infected and a surgical incision was per-formedtodrainthecopiouspus.systemicantibiot-icadministrationtopreventstaphylococcusaureusinfectionwasinitiatedandantisepticdressingwasapplied locally. despite this protocol, a loss ofmusculartissue(figure12a,c)wasevidentduetoanextendednecroticareawhich involveda largeportionofthecalf.ninemonthsaftertheaccident,the surgeon recommended ozone treatment asa last resort to save the leg.after 35 o3-MaHttreatments the results depicted in figure 12b,dwereachieved.tensubsequenttreatmentsdidnotyield any further significant results.this allowedthepatienttoundergoasuccessfulskinautologousgrafttransplantation.

Case 13. a 70-year-old moderately obese dia-betic man underwent surgical excision of a cuta-neous tumouron the foot twoyearsprior toourintervention. the surgical wound only partiallyhealeddespitevarioustreatments,includingtopicalapplicationofautologousplatelet-derivedgrowthfactors. figure 13a, taken before commencingozone therapy,depicts theextentofhealingafter

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figure14Case 14.

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figure15Case 15.

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figure16Case 16.

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figure17Case 17.

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surgeonswerepessimisticanddismantlingof theimplant was seriously considered. the daughter,an anaesthesia nurse, forced her father and thesurgeonstoattemptozonetherapyasalastresort.Majorozonatedautohaemotherapytreatmentwasinitiated and after 30 o3-MaHt treatments theresultsobtainedareshowninfigure17b.thetra-cheostomytubewasremoved.thenecrotictissueandacompleteevenifretractingcicatrizationwasobtained. we considered it imperative to reportthiscasetosupporttheefficacyofo3-Mahtnotonly for the treatment of leg ulcerations, but forany lesion where there is an imbalance betweenmetabolicdeliveryanddemand.

Conclusions

thecaseswehavepresentedare themost sig-nificant and impressive in our experience witho3-Maht. several other patients with lesionsless advanced than those presented in this paperhavebenefittedfromozonetreatments.accordingto the extremely favourable cost/efficacy ratio,we support the application of o3-Maht at leastin those patients suffering chronic ulcers poorlyresponsive to traditional medical and surgicaltherapy.

with success. subsequently, due to chronicizationofthelesion,heunderwentautologousskingraft-ing twice.sixmonthsafter the lastoperation theulceration reappeared and the plastic surgeondecided to follow a conservative approach withtissuegrowthstimulatingdressings.

a few months later, a decision was made toimplement the treatment with o3-Maht. figure15awastakenaftereightmonthsofwoundopen-ing.figure15bdepictstheresultobtainedafter20 sessionsofo3-MaHt,11monthsaftertheappear-anceofthelesion.

Case 16.an82-year-oldwomanwithnoreport-ed concomitant pathologies. over the past threeyears, she has presented an extended chroniculcerationonthelateralaspectofherrightleg.sheunderwentregularmedicateddressingsandsurgi-cal curettage, but the maximum results achievedare shown in figure 16a. she decided to imple-ment ozone treatment as part of her therapy.figure 16b shows the result obtained after 22 o3-MaHttreatments.

Case 17.thelastcasewepresentheredoesnotconcernlegulcerationsbutafailingmusculo-cuta-neousfreeflapimplantedafteroncologiclarynge-ctomyina71-year-oldman.figure17adepictstheclinical aspect three months after the operation.

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99


Effects of Ozone Therapy onOxidative Stress Biomarkersin Coronary Artery Disease Patients

L.deLgado-roche1,g.Martínez-sánchez2,a.díaz-batIsta3,L.re2,4

1 Center of Studies for Research and Biological evaluations, pharmacy and Food Sciences College, University of Havana; Havana, Cuba2 Medinat srl Clinic; Camerano, Italy3 National Institute of angiology and Vascular Surgery; Havana, Cuba 4 pharmacology, D.I.S.M.a.R., University of ancona; ancona, Italy

Key words:ozonetherapy,coronaryarterydisease,oxidativestress

SUMMARY - Coronary artery disease is considered a major cause of death in the western world and its primary pathological manifestation is myocardial damage due to ischemia-reperfusion phenomena. ozone has been used as a therapeutic agent and beneficial effects against the damage induced by renal or hepatic ischemia-reperfusion have been observed in vivo. the present study evaluated the behaviour of oxidative stress biomarkers in coronary artery disease patients after 20 sessions of ozone (50 µg/mL ; 200 mL) by rectal insufflation. Blood samples of 40 patients and 50 healthy subjects were tested by spectro-photometric techniques. Indicators of biomolecular damage, enzymatic antioxidant activity and total anti-oxidant status were determined. We demonstrated that ozone therapy reduced the oxidative stress index as reflected by the increase in superoxide dismutase and catalase activities, the reduction of malondialdehyde, total hydroperoxides and advanced oxidation protein products concentration. Furthermore, an increase in glutathione levels was noted. the results of the present study show that repeated administrations of ozone in non-toxic doses play a positive role in the control of antioxidant/pro-oxidant balance in coronary artery disease patients.

Abbreviations:cad: coronaryarterydiseaseIr: ischemia-reperfusionros: reactiveoxygenspeciessod: superoxidedismutasecat: catalaseh2o2: hydrogenperoxideo2

•-: superoxideanionradicalMda: malondialdehydegsh: glutathionepp: peroxidationpotentialth: totalhydroperoxidesaopp:advancedoxidationproteinproductsgpx: glutathioneperoxidase

Introduction

coronaryarterydisease(cad)remainsamajorcause of death in the western world 1. Myocardialischemia-reperfusion (Ir) injury is the principalcontributor to themorbidityandmortalityassoci-atedwithcad2.

the level of Ir-induced myocardial injury canrangefromasmallinsultresultinginlimitedmyo-cardial damage to a large injury culminating incardiomyocyte death 3. despite the complexity of

the mechanisms responsible for the Ir-inducedmyocardial damage, essential factors leading tocellular injury have been delineated. evidenceindicates that several interrelated factors, includ-ingadecreaseincellularatplevels,accumulationof hydrogen ions 4, production of reactive oxygenspecies (ros) 5, calcium overload 6, and leukocyteactivation,contributetoIrinjury7.

the production of ros during both ischemiaandreperfusionhasbeenconfirmedusingelectronparamagnetic resonance 5 and experimental evi-dencehasaddressedtheroleofrosinmyocardialIr injury 7-9. given the worldwide prevalence ofcadandtheassociatedIr-inducedcardiacinjury,theevaluationofstrategies toprovidecardiopro-tection is an important research area. recently,emphasisandattentionhavefocusedontheuseofmedicalozone10.

In the light of more recent pharmacologicalknowledge, ozone can be considered a pro-drugwhichatcertainnon-toxicdosescaninducearear-rangement of the biochemical pathways with theactivationofasecondmessengerinacascadewithamultiplesystemaction11.

100

effects of ozone therapy on oxidative Stress Biomarkers in Coronary artery Disease patients L. Delgado-Roche

gasmixture(o2+o3).theozoneconcentrationwasmeasuredbyusingabuilt-inuvspectrophotom-etersetat254nm.thepatientsweretreatedwith200 mL of o2-o3 containing 50 µg/mL of ozoneonceadayfor20days.

blood samples for biochemical determinationswere obtained after 12 h overnight fast, at thebeginningofthestudy,and24hafterthelastozoneadministration. these samples were immediatelycentrifugedat3000g,at4°cfor10min.theserumwas collected and aliquots were stored at –70 °cuntilanalysis.

Biochemical determinations

all biochemical parameters were deter-mined by spectrophotometric methods using apharmacia 1000 spectrophotometer (pharmaciaLkb, uppsala, sweden) and a microplate reader(suMa,centerofImmunoassay,havana,cuba).superoxide dismutase (sod) activity was evalu-ated by using randox Ltd. kit cat. no. sd125(diamondroad,crumlin,uk).

themethodemploysxanthineandxanthineoxi-dasetogeneratethesuperoxideradical(o2

•-),whichreacts with 2-(4-iodophenyl)-3-(4-nitrophenol)-5-phenyltetrazolium chloride (Int) to form a redformazandye.sodactivitywasmeasuredbytheinhibitiondegreeofthisreaction16.

catalase(cat)activitywasdeterminedbyfol-lowing the decomposition of hydrogen peroxide(h2o2)at240nmat10sintervalsforoneminute

17.afterprecipitationofthiolproteins,thereducedglutathione (gsh) levels were measured accord-ing to the method of sedlak and Lindsay 18 withellman's reagent (5,5'dithiobis-2-nitrobenzoicacid) 10-2 M (sigma st Louis, Mo, usa), theabsorbancewasmeasuredat412nm.

purifiedgsh(sigmastLouis,Mo,usa)wasused to generate standard curves. the advancedoxidation protein products (aopp) were meas-uredasdescribedpreviously19.

briefly, the technique consists in treating 100µLofseruminpbs(1mL)with50µLofpotas-sium iodide 1.16 M followed by 100 µL of aceticacid.theabsorbanceof thereactionmixturewasimmediatelyreadat340nm.aoppconcentrationswereexpressedasµMofchloramine-t(sigmastLouis,Mo,usa).

concentrationofmalondialdehyde(Mda)wasdeterminedusing theLpo-586kitobtainedfromcalbiochem(Lajolla,ca,usa).Intheassay,theproductionofa stablechromophoreafter40minof incubation at 45 ºc was measured at a wave-lengthof586nm.forstandards, freshlypreparedsolutionsofmalondialdehydebis[dimethylacetal]

evidencethatantioxidantenzymes,nitricoxidepathwaysandothersubcellularactivitiescouldbemodulatedbylowozonedosesisnowprovenandcouldsupporttheeffectsofozoneinmanypatho-logicalconditionssuchasdiabetesmellitus,hepaticandrenalIranddischernia10-14.

the present work considered that treatmentwithozoneatrepeatedlowdosesincadpatientscould reduce the oxidative stress and provide anantioxidantstatus,whichcanservetocompensatethe cad-derived cardiovascular complicationsand also to enhance patients’ quality of life. ourresults confirmed this hypothesis and demon-strated that ozone compensated the antioxidant/pro-oxidantbalanceincadpatients.


Study design

the clinical study was reviewed and approvedby both the national Institute of angiologyand vascular surgery and the pharmacy andfood sciences college (university of havana)committees for research on human subjects,and the procedures were in accordance withprinciples of the declaration of helsinki 15.

all patients gave their informed consentto be enrolled after receiving adequate infor-mation about the study (characteristics of thestudy, benefits and possible side-effects).

Medicalpersonnelwereinstructedtoreportalladverseexperienceswhetherornotdescribedforthemedicationused.adultpatientsofbothgendersand different ethnicity with a diagnosis of cadwhoattendedthenationalInstituteofangiology(havana, cuba) from october 2009 to october2010 were eligible to participate in the study.

exclusion criteria were: severe septic condi-tions, hypersensitivity to the medication to beused, hepatic dysfunction, renal failure (serumcreatinine level >1.32 µmol/L), pregnancy, canceror other serious disease, inability to cooperatewiththerequirementsofthestudy,recenthistoryofalcoholordrugabuse,currenttherapywithanyimmunosuppressive agentoranticonvulsant, con-current participation in another clinical study, orcurrenttreatmentwithaninvestigationaldrug.

thestudyincluded50gender-andage-matchedhealthysubjects.

ozonewasgeneratedbyozoMedequipmentmanufactured by the ozone research center(havana, cuba) and was administered by rectalinsufflation. ozone was obtained from medicalgrade oxygen, and was used immediately upongenerationandrepresentedonlyabout3%ofthe

101


Results

general characteristics of subjects involved in the study

Inrelationtothebaselinecharacteristics(table1), both groups were similar at randomization(p>0.05).Morethan60%ofsubjectsinbothgroupswereolderthan60yearsandmaleswerethemajor-ity.themedicalhistoryofpatientswas character-ized mainly by cad, but also by hypertension,ischemiccardiopathyandmyocardialstroke.

patients presented a prevalence of risk factorsforcardiovasculardiseases(cvd)suchashyperc-holesterolemiaandobesity.

the conventional treatments were those usedto control hypertension (captopril in 40%, nitro-surbide in 45%, and atenolol in 60% of patients,respectively), hypercholesterolemia (ateromixol®in70%ofpatients),andcvd(aspirin®in90%ofpatients).

Biomarkers of oxidative stress

table2showsthebehaviourofoxidativestressbiomarkers.after20sessionsofozonetreatment,th levels in cad patients were similar to thecontrolgroup.

atthebeginningofthestudy,Mdaconcentra-tion was higher in patients (p<0.001) compared

(sigma st Louis, Mo, usa) were employed andassayedunderidenticalconditions20,21.

Quantification of total hydroperoxides (th)was measured by bioxytech h2o2-560 kit (oxisInternationalInc.,portland,or,usa).

the assay is based on the oxidation of fe2+ tofe3+ by hydroperoxides under acidic conditions.ferricionsbindwiththeindicatorxylenolorange(3,3'-bis(n,n-di(carboxymethyl)-aminomethyl)-o-cresolsulfone-phtalein, sodium salt) to form astable colored complex, which can be measuredat560nm.

finally, todetermine susceptibility to lipidper-oxidationtheperoxidationpotential(pp)wascal-culated.sampleswereincubatedwithasolutionofcoppersulfate(2mM)at37ºcfor24handthenMdaconcentrationwasdetermined22.

Statistical analysis

the outLIers preliminary test for detec-tion of error values was initially applied. datawere subsequently analyzed by one-way analysisof variance (anova) followed by a homogene-ity variance test (bartlett-box). student's t-test(two-tailed) was used to determine differencesbetweengroups.datawereexpressedasthemean±standarddeviation(sd).the levelofstatisticalsignificance employed was at least p<0.05 for alldeterminations.

Table 1 baselinecharacteristicsofcontrolsubjectsandcadpatients.

Characteristics Control group(n=50)

CaD patients(n=40)

n % n %

age(years)

50-6061-7071-80>80

1123197

18.0038.0032.0012.00

122071

30.0050.0017.502.50

gender femaleMale

1931

38.0062.00

1426

35.0065.00

History cadhypertensiona

MyocardialstrokeIschemiccardiopathy

––––

4032337

100.0080.007.5092.50

Risk factors hypertensionhypercholesterolemiab

obesityc

smoking

––––

32141721

80.0035.0042.5052.50

Complementarydiagnostic criteria

tc(mM)bMI(kg/m2)

3.18±0.5022.10±2.30

6.8±1.3134.5±9.67

Legend: ahypertensionwasdefinedaselevationofsystolic(>140mmhg)and/ordiastolic(>90mmhg)bloodpressure. bhypercholesterolemia:increaseintotalcholesterol>6.7mM.bMI,bodymassindex:weight(kg)/height(m2).nosignificantstatisticaldifferencesbetweengroups(p>0.05)werefoundforageandgender.cad:coronaryarterydisease.tc:totalcholesterol.

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Discussion

In addition to traditional risk factors for cvd(hta, obesity, hyperlipidemia, diabetes, ciga-rettesmokingandage),oxidativestress(os)andinflammationarenowbeingconsideredsignificantriskfactorsforcvdandotherdiseases.underosconditions take place a disruption of redox bal-ance,affecting theantioxidantstatusand favoredtherosgeneration23,24.

ozone therapy is capturing attention all overtheworldsincethebasicstudiesclarifiedthemainbiochemical mechanisms of action and the realpossibilityoftamingozonetoxicity.therearegoodreasonstobelievethatthetherapeuticefficacyofozone therapyconsists in simultaneously improv-ing circulation and oxygen delivery, in enhanc-ing the release of autacoids, growth factors andcytokines and in reducing chronic os 10.this lasteffect has been approached in the treatment ofmanydiseases,suchasdiabetes13,hepaticischemia/reperfusioninjury14andothers.

anefficientclinicaldiagnosticoftheredoxbal-anceincadpatientsisofparamountimportanceto control the degenerative damage associatedwithos,ortomonitoringtheeffectofanutritionalor therapeutic regimen. In order to establish thebehaviourofantioxidantstatusafterozonethera-pywedeterminedtheserumactivityofsodandcatenzymesandalsothenonenzymaticantioxi-dantsgshandwatersolublereducedsubstances.

sods are metal-containing proteins that cata-lyze the removal of superoxide anion, gener-ating h2o2

25. then, this ros is converted in

to healthy individuals, but after the last ozoneadministrationthisparameterwaspositivelymodi-fied. Mda levels were reduced in cad patientscomparedtotimezero,withstatisticaldifferences(p<0.05).

similarbehaviourwasobservedforaoppcon-centrations. In patients, ozone therapy induced asignificant reduction ofaopp levels (p<0.05) incomparison to the high levels determined beforethegasadministration.

In relation to the antioxidant enzyme activity,bothcatandsodshowedasignificantincreasein cad patients (p<0.001) compared to controlgroup.ozonetreatmentpositivelymodulatedtheactivityoftheseenzymes.attheendofthestudythe enzyme activity was significantly lower thanvaluesattimezerointhesamepatients(p<0.001).the concentration of low weight molecular anti-oxidant gsh in cad patients was significantlylower compared to reference values in controlgroup (p<0.001). It was important to note thatthe ozone administration promoted a significantincrease(p<0.05)ofgshinpatientsafter20ses-sionsoftreatment.

In order to measure the susceptibility to lipidperoxidation, as indicator of total antioxidantactivity,theppwasdetermined.

theresults showedahighlyoxidativedamageon lipids in patients with cad respect to con-trol group, with statistical differences (p<0.001).however, after ozone treatment, the antioxidantstatus of cad patients was higher than in timezero,asreflectedthemeasuredppandbacktothenormalinterval.

Table 2 effectsofozonetherapyonbiochemicalparametersofoxidativestress.

oxidative stressbiomarkers Control group (n=50)

CaD patients (n=40)

time 0(Conventional

therapy)

20 days afterozone therapy

Mda(µM) 3.87±0.91a 14.78±5.03b 9.21±3.30c

th(µM) 63.00±8.09a 92.96±7.70b 65.45±6.52a

aopp(µMofchloramines) 9.19±0.64a 25.63±6.93b 15.84±3.34b

gsh(mmol/L) 789.45±97.30a 378.73±54.12b 517.17±81.36c

cat(u/L/min) 231.80±11.33a 804.70±108.67b 584.10±94.77c

sod(u/mL/min) 11.35±1.97a 39.75±8.86b 22.17±12.36c

pp(µMofMda) 9.17±0.82a 18.93±6.50b 8.48±1.73a

thetableshowsthemeans±sdofallmeasuredbiomarkers.attimezerothepatientsonlyreceivedtheirconventionalmedicationforthecad,duringtheexperiment,patientsreceivedtheozonetherapyconcomitantlywiththeconventionalmedication.differentlettersrepresentstatisticaldifferences(p<0.05)betweenthesameset.

103


only as an indicator of os, but also as a markerof progression of atherothrombotic risk in cadpatients 37.also, total hydroperoxides levels werereducedafterozonetreatmentwhichsuggeststhatlipid peroxidation was positively modulated byo2-o3.

aminoacids,peptides,andproteinsarevulner-abletotheattackofavarietyofrosandrelatedoxidants.accordingtoourresultsitisevidentthatthehighaopplevelsinthecadpatientsplasmacouldbe reducedbyozone therapy.the trendofthisparametermediatedbyozoneadministrationisverypositivebecauseisknowthattheoxidativedamage on proteins is correlated with the extentof atherosclerotic lesions and the occurrence ofadversecardiovascularevents24.

as an expression of the total antioxidant-oxi-dantbalance,theppwaspositivelyaffectedattheendofthestudyrespecttothebeginninginthosepatients who received the ozone therapy. a sig-nificantdecreasecomparedtotheinitiallevelwasfoundafterozoneadministration.thefactthatthevalues return to normal intervals can be consid-eredanormalizationoftheredoxstatus.

Conclusions

repeated rectal insufflations of ozone con-tributed to enhance the activity of antioxidantenzymes and non enzymatic defenses. In addi-tion, the treatment reduces the oxidative injuryof lipidsandproteins.theseobservationssuggestthat ozone may be used in combination with theconventional drugs for cad. consecutive andspacedtreatments,inaregularform,arenecessarytoobtainaneffectiveantioxidantactivationstatusincadpatients.normalizationoftheantioxidant/ pro-oxidant status in these patients could be apositive influence to avoid future complicationsandfatalevents.futureclinicaltrialswillbeneces-sarytoestablishhowlongtheantioxidantstatusismaintainedaftertherapyandhowoftenitwillbenecessarytorepeattheozonetreatment.

Acknowledgements

the study was partially supported by the National Institute of angiology and Vascular Surgery, Havana, Cuba.

water and oxygen in a cat-catalyzed reaction

26,27. In the present study ozone treatment pro-ducedanincreaseinantioxidantenzymesactivity.overexpressionofsodandcatinculturedcellsandanimalmodelshasprovidedprotectionagainstthedeleteriouseffectsofawiderangeofospara-digms27.theseobservationssuggestthatthetreat-mentwith therapeuticagents,whichpromote theexpression of endogenous antioxidants, representa plausible alternative in the treatment of thoseos-mediateddiseasessuchascad28.

the sod and cat activity trends observedin the present work could be associated with theobservations that ozone treatment may promotea moderate os which, in turn, increases antioxi-dant endogenous systems protecting against oxi-dativedamage10,12. It isknownthat theprotectivemechanismofozonemay involveproteinsynthe-sis 14. elevated ros concentrations induce geneexpression in many cells, whose products exhibitantioxidantactivity.amajormechanismofredoxhomeostasisisbasedontheros-mediatedinduc-tionofredox-sensitivesignalcascadesthatleadtoincreasedexpressionofantioxidants29,30.

anotherassayedcomponentoftheendogenousantioxidant system was gsh.the ozone therapypositively modulated the gsh level in patients.gshplaysanimportantroleinmediatingseveralredox-based signaling processes as well as geneexpressionwithinthecell31.gshisalsoessentialfortheactivityofthepowerfulglutathioneperoxi-dase(gpx)antioxidantenzymefamily32,33,inanti-oxidantgenetranscription33andintransthiolationreactions 34. the importance of gsh in vascularpathologywashighlightedbystudiesshowingthatit protects the macrophages from oxidized low-density lipoprotein (ox-LdL)-induced cell injurywithintheatheroscleroticlesion35.

biomarkers of ros-induced damage have thepotential not only to determine the extent ofoxidative injury, but also to predict the poten-tial efficiency of therapeutic strategies aimed atreducingsuchanos36.Inlinewiththeincreaseinantioxidantenzymestherewasareductionofbio-moleculesdamages.althoughMdalevelsincadpatients were higher than in control group, wasvery satisfactory the result after the ozone treat-ment,becauseattheendofthestudytherewasasignificantlyreductionofthisend-productoflipidperoxidation.IthasbeenreportedthattheplasmaMda-lowdensitylipoproteinlevelsareusefulnot

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13 Martínez-sánchez g,al-dalain sM, Menéndez s, et al.ozone treatment reducesbloodoxidative stressandpan-creasdamage ina streptozotocin-induceddiabetesmodelinrat.actafarmbonaerense.2005;24:491-497.

14 ajamieh hh, berlanga j, Merino n, et al. role of pro-tein synthesis in the protection conferred by ozone-oxi-dative-preconditioning in hepatic ischemia-reperfusion.transplantInt.2005;18:604-612.

15 wMa.world medical association declaration of helsinki.ethical principles for medical research involving humansubjects.adopted by the 18thwMa generalassembly,helsinki, finland, june 1964: j Int bioethique. 2004; 15:124-129.

16 boehringer M. biochemical information.a revised bio-chemical reference source. In: enzymes for routine.germany:boehringerMannheim;1987.p.15-16.

17 haining jL, Legan js. Improved assay for catalase baseduponsteady-statesubstrateconcentration.analbiochem.1972;45:469-479.

18 sedlak j, Lindsay rh. estimation of total protein boundand non protein sulfhydryl group in tissue with ellman`sreagent.analbiochem.1968;25:192-205.

19 witko-sarsat v, friedlander M, nguyen-khoa t, et al.advanced oxidation protein products as novel mediatorsofinflammationandmonocytesactivationinchronicrenalfailure.jImmunol.1998;161:2524-2532.

20 erdelmeier I,gerardd,yadanjc,etal.reactionsofn-methyl-2-phenyl-indole with malondialdehyde and 4-hy-droxy-alkenals. Mechanistic aspects of the colorimetricassay of lipid peroxidation. chem restoxicol. 1998; 11:1184-1194.

Livandelgadoroche,Mdcalle222y27ano.21425cp13600,Lacoronela,LaLisaLahabana,cuba.tel.:(53-7)2719531fax:(53-7)2736811.e-mail: [email protected] [email protected]

105


Oxygen-Ozone Treatment of Verrucas

t.gastaLdICentre Médical de la Côte; Corcelles Ne, Switzerland

Key words:oxigen-ozone,verrucas

SUMMARY -this paper describes the oxygen-ozone treatment of verrucas using bags and washing with ozonized water resulting in a complete clinical resolution.

Introduction

verrucasareskinwartscausedbyhumanpapil-lomavirus(hpv)belongingtothepapovaviridaefamily. they are benign lesions made up of anucleusofskinstriationsfedbybloodvesselsandcoveredindifferentlayersofepithelialtissue.thevirus penetrates the epidermis causing infectionandtriggeringexcessiveskinreplication.verrucasare spread by surface contact (the virus remainsin the skin and is not found in blood) usually incommunalareaslikeshowers,gymsandswimmingpoolswherethewarmatmosphereandmoistsur-faces favour survival of the virus in active form.withouttheseenvironmentalconditionstheviruswillnotsurvivelongoutsidetheskin.

verrucas can develop on any part of the bodybutoftenonlyaffectspecificareas,e.g.hands,feet,elbowsandknees.thesepartsoftheskinarethosemostcommonlysubjecttomechanicaltraumaandcontactwith theexternal environmentand thosemost likely to have microlesions the virus canattack.

the appearance of verrucas varies dependingon the body area affected and the virus straininvolved. verrucas can be divided into common(orvulgar)verrucas,andplantar,flatandfiliformverrucas.commonverrucashavethesamecolourastheskinandareusuallydistinguishedbytheirtypical rough often crinkled surface. plantar ver-rucas are confined to the soles of the feet andusually alter the skin striae. for this reason andthefactthatverrucasmayalsocontaintinyblackpetechiae,plantarwartsare readilydistinguishedfromcallusesandcorns.theblackpetechiaerep-resent tiny haemorrhages under the skin formedby stretching the papillae. verrucas of this typetend to be soft, flat and covered by calluses andtend to be painful on application of pressure onwalking.


betweenjune2005andapril2011wetreated55patientspresentingverrucasondifferentpartsofthebody:45%onthehands,40%onthefeetand15%withlesionselsewhere.

twomethodswereused:1. Intralesionalinjectionofanoxygen-ozonemix-tureataconcentrationof40micrograms/mlusing5 ml syringes and 27 or 30 g needles. Injectionswere administered every three to four days untiltheverrucasdisappeared.2. washingwithozonizedwaterataconcentrationof25micrograms/mlassociatedwithbagscontain-ingoxygen-ozoneataconcentrationof50micro-grams/ml.treatmentswererepeatedeverythreetofourdaysuntiltheverrucasdisappeared.

Results

thirty-five patients underwent treatment withozonizedwaterandbagswhile20receivedoxygen-ozoneinjections.

all patients fully recovered with a disappear-anceof theverrucas inaperiod ranging from14to 35 days (average 21, average 17 with oxygen-ozoneinjections,23withwaterandbagtreatment).Multipleverrucaswerepresentin41patients.

the verrucas regressed progressively and ondetachmentnoscarringoccurred.bothtechniqueswere well-accepted by the patients as they arecompletelypainless.

Conclusions

to date, human papilloma virus has not beenthe topic of methodical in-depth studies. forthis reason a treatment guaranteeing complete

106

oxygen-ozone treatment of Verrucas t. gastaldi

figure5 figure6

figure3 figure4

figure1 figure2

107


figure7 figure8

figure9 figure10

figure11 figure12

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oxygen-ozone treatment of Verrucas t. gastaldi

Reference

1 gibbss,harveyI.topicaltreatmentsforcutaneouswarts.cochranedatabasesystrev.2006;3:cd001781.

cure was not available until recently. Leadingmedical journals1 claim that no treatmenthas a likelihood of success greater than 73%.thetreatmentsutilizedtodateinclude:– surgicalresection.– cryosurgery.– keratolyticchemicals.– Intralesional injections: consisting in the injec-

tionofinterferonintotheverruca.– Lasersurgery.– vitamineapplication.– figtreelatex.

amongthemanytreatmentsproposed,oxygen-ozonetherapyyieldedparticularlysignificantout-comeswithoutscarringaftertherapyorpaindur-ing treatment. no patients suspended treatment.oxygen-ozonetreatmentisaninnovativetherapyrecommendedforitssimplicityandsuccessrates.

tizianogastaldi,MdcentreMédicaldelacôteruedepetit-berne142035corcellesnetel.:0327271100fax:0327271101e-mail:[email protected]

figure13 figure14

figure15

109

www.centauro.it International Journal of ozone therapy 10:109,2011

A New Infiltrative Paravertebral Approach for the Cure of Disc Herniation Using O2/O3

r.vIgLIoLIBrescia, Italia

Key words:oxygen-ozone,paraventricular,discherniation

SUMMARY - this paper describes a new infiltrative paravertebral approach for the cure of disc herniation using o2-o3.

Introduction

ahighnumberofpatientsapplyfor infiltrativeparavertebraltherapy,or‘complaince’forsciaticaand/or severe cervicobrachialgia given the crip-plingeffectofthepathology.however,itisstillthedoctor’sdutytotrytoreducetheemotiveimpactcausedbythepainoftheevent(evenifitsinvasivenature is limited),maintainingoreven improvingthefavourableoutcomesofthismethod.


atthechirotherapeuticcenterinbrescia,Italyapproximately 850 patients have undergone thisnew operative model over the last two years.the treatment protocol consists in paravertebralmonofiltrationofthesideoftheherniaorprotru-sivelesion,indicatedbyclearradialsymptomology,usinga0.4×40mmneedleandinjecting4/5mlofo2-o3gasatanozoneconcentrationof-30mcrgtoadepthofo3-ml.o2anotherfollowedby2-3mltothemoresurfacemuscleforatotalofsixtoeightsessionsatthreeweekintervals.withnoclearradi-al symptoms, the scheme is identical, being surehowevertoalternatetheinfiltrativesite(left-right)ateachsessionandpossiblyalsoincreasethetotalnumberofsessionstoeightorten.thepatientcanbeallowedtostandimmediatelyafterthebandageisputinplace,soastoreducetheclino-orthostatictime,whichwilldrastically reduceposturalhypo-tensionwhichoccursaboveallinelderlypatients.

Results

thismethodhasbeencarriedoutfortwoyearswith very consistent results. patients’ sympto-mological improvement, meaning a reduction of

at least 50% of the vas score, was achieved inapproximately80%ofpatients,inlinewithprevi-ousfindings.therewasalsoacompleteabsenceofadverseside-effects.

Conclusions

the shorter ‘therapeutic time’, low traumaticnature of the method together with the brevityof the cycle (maximum therapy three weeks) allcontribute to lowering anxiety in patients and atthe same time making the o2-o3 therapy saferandmorepracticalwhilestillmaintainingitswell-established therapeutic effects. this approach isconsiderably less invasive than traditional treat-mentmethods,partiallyduetothethinnerneedles,andthesmallervolumeofgasusedineachsession.however,itiscloselylinkedtotheuseofneedlesat least 40 mm long and three weekly treatmentsessions.

drriccardoviglioliviaIndipendenza5/abrescia-Italye-mail:[email protected]

110

Letters to the editor

Letters to the Editor

drMatteobonettiscientificdirectorInternationaljournalofozonetherapy

Caro Direttore

Lainformodiaverricevutoneigiorniscorsila sentenza del tas di Losanna, dove sonostato ascoltato come esperto- Mi sembraopportunodarespazioaquestainformazione,importantissima per chi ha creduto e credenell’ozono terapia. Informazione nella qualesiattribuisceilgiustoriconoscimentopubblicoall’usomedicaledell’ozono.

Lamberto Re www.lambertore.com

trIbunaLearbItraLedeLLosport

arbitrounico:prof. Ulrich Haals, Zurigo, Svizzera

omissis

9.31. tornando sul punto concernenteI’adeguatezza del ricorso all’ossigeno-ozonoterapia per la cura delle patologie soffertedagliatleti,bisognaconstatareche,anchequi,nonvièconcordanzadiopinionitragliespertidi parte. ed infatti, mentre il consulente tec-nicodelricorrenteha fermamente sostenutol’opportunità dell’impiego della terapia pertali patologie, gli esperti dell’upa·conI sisono mostrati decisamente scettici al riguar-do. anche il consulente tecnico d’ufficio haespresso la sua opinione sul punto, rilevandochepiùcheescludersicategoricamentel’utilitàdella terapia per la cura delle patologie indi-cate,sipuòconstatarechenonvisonodeidati

scientificichemostrino,inmanierainequivoca,I’efficaciadellastessa.

9.32.orbene,mentre,riscontrandounadivi-sionediopinioniqualequellaappenadescritta,sembraopportunoevitarediemettereverdettiscientifici sulla questione dell’ efficacia dellaterapiaperiltrattamentodellepatologieindi-cate,parepotersiaffermareche,perI’indaginevoltaadesaminarel’intentodelricorrente,visiaunaltroelementochepuòritenersiessererilevante, al di la della reale efficacia dellaterapia,infatti,deveconstatarsicheessavieneeffettivamente impiegata dai medici che nesostengono l’utilità, per il trattamento dellepatologie allegate. Questo dato -che si evincedalle perizie prodotte dal ricorrente e dalledichiarazionirilasciatedalprof.reinudienza-non è stato smentito dai consulenti tecnicidell’upa-conIche,comedetto,sisonolimi-tati a mettere in discussione l’efficacia dellaterapia.e’possibile,dunque,cheilricorrentefosserealmenteconvintodell’utilitàdellatera-pia nei casi in cui essa sia stata praticata. suquesta base, allora, non sembra che il fattocheilricorrenteabbiautilizzatolaterapia inoggettoper la curadellepatologiedi cui sof-frivanogliatleti,chealuisieranorivolti,possaessere interpretato automaticamente comeun’intenzionedisottoporreglistessiapratichedopanti.

omissis

Losanna,2agosto2011

Federazione Italiana diOSSIGENO-OZONOTERAPIA

FEDERAZIONE NAZIONALEORDINE VETERINARI ITALIANI

Roma, 13 Aprile 2011

Mariastella Gelmini

Prof. Matteo BonettiBrescia

Egregio Professore, desidero innanzitutto esprimere i miei sinceri apprezzamenti per la promozione del 3° Convegno Internazionale di Ozonoterapia che in questi ultimi anni ha trovato sempre più ampia ed efficace applicazione in campo medico. La veste internazionale dell’iniziativa che ha un ruolo di alta rilevanza scientifica, la presenza di autorevoli relatori saranno sicuramente motivo di apporto di esperienze concrete e convalidate e sono pertanto spiacente di non poter essere presente a causa di inderogabili impegni di Governo. Nel ringraziare per il cortese invito, invio un cordiale saluto e formulo i migliori auguri per il successo dell’importante evento.


FEDERAZIONE NAZIONALEORDINE VETERINARI ITALIANI

Dott. CARMELO SCARCELLADirettore Generale ASL di Brescia

Dott. MARCO TOMAin rappresentanza del Sindacodi Brescia Dott. Adriano Paroli

Dott. RAFFAELLO MANCINIPresidente Ordine dei Medici

Dott. ENZO MONTEMAGNOQuestore di Brescia

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Dott. CARMELO SCARCELLADirettore Generale ASL di Brescia

Dott. MARCO TOMAin rappresentanza del Sindacodi Brescia Dott. Adriano Paroli

Dott. RAFFAELLO MANCINIPresidente Ordine dei Medici

Dott. ENZO MONTEMAGNOQuestore di Brescia

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The Francesco Riccardo Monti prize for life time achievements is a recogni-tion for scientific work done to spread Oxygen-Ozone Therapy practice in Italy and around the world.

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Professor Vjiay Kumar (India), a Neurosur-geon and President of the World Federation of Oxygen-Ozone Therapy has the impor-tant merit of having spread the practice of oxygen-ozone therapy with scientific rigour throughout Asia starting from his native city New Delhi. Vjiay Kumar is currently a world reference for the treatment of disc hernia-tion.

Professor Vjiay Kumar

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Professor Marco Leonardi

Professor Marco Leonardi (Italy) is full pro-fessor of Neuroradiology at Bologna Uni-versity, President of the Italian Federation of Oxygen-Ozone Therapy and founder of the Italian Journal of Oxygen-Ozone Therapy, currently the International Journal of Ozone Therapy, a journal indexed in Elsevier’s Em-base database and internationally acknow-ledged as the only scientific journal devoted to ozone therapy.

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Professor Vjiay Kumar (India), a Neurosur-geon and President of the World Federation of Oxygen-Ozone Therapy has the impor-tant merit of having spread the practice of oxygen-ozone therapy with scientific rigour throughout Asia starting from his native city New Delhi. Vjiay Kumar is currently a world reference for the treatment of disc hernia-tion.

Professor Vjiay Kumar

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Professor He Xiaofung

Professor He Xiaofung (China) is head of the Interventional Radiology Service at Guangzhou University and President of the Chinese Society of Ozone Therapy. His prize is awarded for his research activity, mainly on animals.

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Professor Velio Bocci

Professor Velio Bocci (Italy) has been head of the Pharmacology Institute at Siena Uni-versity since 1971. He is the single person who has contributed most to international scientific research into oxygen-ozone therapy worldwide. His career has been crowned by more than five hundred articles published in the international literature.

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Professor He Xiaofung

Professor He Xiaofung (China) is head of the Interventional Radiology Service at Guangzhou University and President of the Chinese Society of Ozone Therapy. His prize is awarded for his research activity, mainly on animals.

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2013FIRENZE

II° FRANCESCO RICCARDO MONTIPRIZE

Presidente Gianantonio Pellicanò Firenze (Italy)

II° FRANCESCO RICCARDO MONTI

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2013FIRENZE

II° FRANCESCO RICCARDO MONTIPRIZE

Presidente Gianantonio Pellicanò Firenze (Italy)

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PROF. MARCO LEONARDIUniversity of Bologna, ItalyChair of Neuroradiology - Bellaria Hospital - www.neuroradiologia.unibo.it

Complimenti per il grande successo ottenuto, un Congresso ad altissime livello scientifico. Complimenti ancora Matteo”

Marco

PROF. MASSIMO GALLUCCIProfessor and HeadDept of Neuroradiology, University-Hospital - 67100 L’Aquila - [email protected]

Complimenti per il grande lavoro fatto per questo III° Congresso Mondiale di Ozonoterapia e per il successo ottenuto, un salto di qualità per il mondo-dell’ozonoterapia.

HE XIAOFENG M.D.,PH.D.Nanfang Hospital, Southern Medical UniversityGuangzhou, China

I, on behalf of the Chinese Society of Ozone Therapy, warmly congratulate the holding of the Third World Congress of Oxygen-Ozone Therapy, and wish the conference a complete success, and wish the delegates good health and hap-piness!

VIKRAM AND VIJAY KUMAR

Dear Prof. Bonetti

Thank you again for the wonderful conference in Brescia. It was a tremendous show! Congratulations to all of you. Thank you.

Warm regardsVikram and Vijay Kumar

ANGELES ERARIO AND ANIBAL GRANGEAT IAOT www.iaot.com.ar

Dear Matteo,thank you very much for inviting me. Congress and the entire organization were perfect and I look forward to participating with my boss in the WFOOT. You can count with us.Once again, it was our pleasure to be part of the conferences A big hug from Buenos Aires, Argentina.

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ALFONSA MARTÍN

Dear colleague,It has been a pleasure meeting you in Brescia.Our Association, the ACEOOT, is very grateful to you for your kind invitation to participate at the Congress, which was most interesting. We wish you luck for your recent joint cooperation agreement with ISCO. We hope to be able to maintain, in future, the kind and respectful relationship we have been having so far.

Sincerely yours

GIANNI PELLICANÒ

Grazie di cuore Matteo, organizzazione perfetta, un grandissimo congresso ad alto livello scientifico, grazie davvero e sentiti orgorglioso del grande lavoro fatto.

YVES BERGERON

Dr. Bonetti,first of all, I want to thank you and congratulate you for the excellent organiza-tion of the congress.It would be interesting to gather together the presentations about the muscu-loskeletal system on one side, and the presentations pertaining to internal me-dicine on the other side.Also, presentations varied a lot on a scientific point of view. Some had high scientific standards whereas others were more personal and less critical about this therapeutic resource.I myself believe it is important to keep an objective, critical mind, about ozone therapy instead of presenting it like a panacea appreciated by some, and little-know by others.I’ll be glad to further discuss this matter with you. Thanks once again.

Regards

DR. MARIAN SIMKOFribourg, Svizzera

Dear Matteo,Thank you very much for the invitation.The meeting, was wonderfull.La ringrazio ancora di tutto e il piacere di incontrarci in futuro.

ALESSIO ZAMBELLO

Caro Matteo,complimenti vivissimi per questo III Congresso Mondiale, so quanto lavoro ti è costato! Un congresso che segna una svolta, per la prima volta si è cambiato passo. Relatori di grande spessore scientifico, professionale ed umano. Relazioni serie, ben preparate, casistiche numerose e proposte terapeutiche razionali. L’ozono sta acquisendo una meritata autorevolezza, la strada è questa.

Un salutoAlessio

RAMIRO ALVARADO

Mi Caro Matteo,Just coming back to my country, I wanto to Thank You for the invitation to participate in the wonderfull congres os Ozone Therapy, really was very nice CONGRATULATION for your big and hard effort I hope to see you again very soon. Let me know about all the plans and programs you have

With My best Regards

STAFA ALTINCaro Matteo,volevo di nuovo ringraziarti e farti i complimenti per il tuo lavoro nell’organizza-zione del Congresso. Un bel Congresso, con sostanza e dettagli curati. Mi è piaciuto molto.

A presto! Altin

LAMBERTO RE, MD

Carissimo Matteo,a distanza di alcuni giorni dal Congresso da te coordinato e splendidamente organizzato, invio queste mie note in merito all’evento.Desidero innanzitutto complimentarmi per l’alto spessore scientifico degli argo-menti trattati che hanno riguardato tutti i campi di applicazione dell’Ozono Tera-pia con approfondimenti tematici che saranno di grande supporto per il futuro di questa nuova branca della Medicina.Forse per la prima volta, e dinanzi ad una platea di elevato impatto internazio-nale, sono stati discussi argomenti e dati scientifici che confermano la crescita di questa terapia e l’impatto che potrà avere nella pratica clinica. Veramente complimenti e auguri per un futuro che, sono certo, riserverà all’Ozono Terapia uno spazio sempre più ampio soprattutto se supportato da Convegni e Aggior-namenti all’altezza di quest’ultimo evento bresciano.

Saluti vivissimi a te e al nostro caro Presidente Marco.Lamberto Re

D.I.S.M.A.R. University of Ancona

Clinical Pharmacology and ToxicologyVia Brecce Bianche - 60128 Ancona (IT)

Mail: [email protected]: www.lambertore.comMobile: + 39 339 5372953

ALFONSA MARTÍN

Dear colleague,It has been a pleasure meeting you in Brescia.Our Association, the ACEOOT, is very grateful to you for your kind invitation to participate at the Congress, which was most interesting. We wish you luck for your recent joint cooperation agreement with ISCO. We hope to be able to maintain, in future, the kind and respectful relationship we have been having so far.

Sincerely yours

GIANNI PELLICANÒ

Grazie di cuore Matteo, organizzazione perfetta, un grandissimo congresso ad alto livello scientifico, grazie davvero e sentiti orgorglioso del grande lavoro fatto.

YVES BERGERON

Dr. Bonetti,first of all, I want to thank you and congratulate you for the excellent organiza-tion of the congress.It would be interesting to gather together the presentations about the muscu-loskeletal system on one side, and the presentations pertaining to internal me-dicine on the other side.Also, presentations varied a lot on a scientific point of view. Some had high scientific standards whereas others were more personal and less critical about this therapeutic resource.I myself believe it is important to keep an objective, critical mind, about ozone therapy instead of presenting it like a panacea appreciated by some, and little-know by others.I’ll be glad to further discuss this matter with you. Thanks once again.

Regards

DR. MARIAN SIMKOFribourg, Svizzera

Dear Matteo,Thank you very much for the invitation.The meeting, was wonderfull.La ringrazio ancora di tutto e il piacere di incontrarci in futuro.

GIULIANO FABRIS

Caro Matteo,

ancora complimenti per l’organizzazione e la riuscita del Congresso Mondiale. La presenza di personalità di indubbia autorevolezza nel mondo dell’ozonote-rapia e la qualità delle relazioni sono state di elevata valenza scientifica. Il 3° Congresso Mondiale di Brescia, è un effettivo e forte punto di riferimento, un confronto delle varie esperienze mondiali, un ulteriore traguardo per l’ozono-terapia, che come è stato confermato è di grande efficacia terapeutica, basso costo ed esiguo rischio nell’ernia del disco e nel dolore muscolo-scheletrico.L’ampia e qualificata partecipazione di vari specialisti in branche mediche e chirurgiche diverse, ma complementari, ha apportato certezze per i medici che si dedicano all’ozonoterapia.

Un grazie per tutto il lavoro che con passione hai svolto.

ANNUNZIATA IZZO E ANTONELLA BERTOLOTTIINTERMED ONLUS

La riuscita del III World Congress of Oxygen-Ozone dell’aprile 2011 a Brescia, è stata eccezionale, soprattutto per la possibilità di confrontarci con i colleghi e di trovare riscontro in alcune scelte che, soprattutto nei Paesi in via di svilup-po, sono a volte difficili. Il nostro punto di vista è stato supportato anche dalla dott.ssa Iabichella,che ci ha suggerito di applicare la TOA anche sull’Ulcera di Buruli. Anche la collega Schwartz Tapia che ha parlato di una patologia che noi trattiamo spesso nei nostri dispensari in Africa, ci ha dato suggerimenti impor-tanti ed utili. Inoltre il trattamento con olio ozonizzato è risultato utilizzabile sulle lesioni che stiamo curando.

JOSIP BURIC

Caro Matteoinnanzi tutto ti volevo ringraziare ancor una volta per avermi dato la possibilità di presentare le mie relazioni Il convegno è stato di alto livello e non lo dico solo per cortesia. Ho incontrato persone di diverse nazionalità il che conferma la internazionalità del convegno e la diffusione del ozono stesso. Nella stragrande maggioranza dei casi del-le presentazioni cui ho avuto modo di assistere devo confessare che non mi aspettavo un livello così elevato di preparazione scientifica ma anche di meto-dologia della ricerca clinica che ha portato alla presentazione stessa. Inoltre, la condotta del intero convegno nella lingua inglese ha dato quel tocco in più al congresso che certamente se lo è meritato. Congratulazioni per l’organizzazio-ne meticolosa e attenta dell’evento. Al prossimo anno.

Sinceri saluti

MARIA LETIZIA IABICHELLA

Il recente consenso mondiale di ossigeno-ozonoterapia ha offerto la possibilità di interscambio di esperienze pluridisciplinari ed al tempo stesso ha stimolato in noi ricercatori la collaborazione nella programmazione di protocolli necessari a documentare ulteriormente i risultati clinici con i dati ottenuti in laboratorio.All’impegno di Matteo Bonetti un encomio di merito.

F. ALBERTINI

Servizio di neuroradiologiaistituto Clinico S.Anna - Brescia

L’utilizzo dell’ozonoterpia nelle varie discipline mediche suscita sempre mag-giore interesse sia nel mondo scientifico che da parte dei pazienti. Grande oc-casione di incontro è stato quest’anno il Congresso Mondiale di Ozonoterpia organizzato a Brescia, con numerose sessioni scientifiche dedicate a tutti i set-tori di impiego e alle novità. L’impatto e il livello scientifico del congresso sono stati notevoli permettendo un utile confronto tra gli specialisti e evidenziando come tale terapia sia spesso in grado di influenzare molti aspetti e la qualità di vita dei pazienti.

ROBERTO DALL’AGLIO

il congresso di Brescia è stato contrassegnato da due grandi elementi: da una parte si è assitito alla sempre piu raffinata definizione delle tecniche interven-tistiche neuroradiologiche e neurochirurgiche dell’ozono sulle patologie del rachide;dall’altra si è osservata una nuova gamma di applicazioni dell’ozono legate all’attivita antalgico-antiinfiammatoria che è stata un primo progresso clinico delle dimostrazioni di interferenza positiva col sistema immunitario di-mostrate in vitro:dalle premesse biologiche e speculative di Bocci e Re, la lectio magistralis di fahmy sulle applicazioni reumatologiche ha segnato una imposta-zione innovativa in questo campo, foriera di promettenti esperienze da espleta-re anche in studi comparativi coi farmaci antireumatici.

UNIVERSITÀDEGLI STUDIDI MILANO

DANIEL ROUBINI

Grazie per lo splendido congresso. Ottima organizzazione con relazioni originali e stimolanti. Ho appreso fatti nuovi e consolidato le mie conoscenze.

GIULIANO FABRIS

Caro Matteo,

ancora complimenti per l’organizzazione e la riuscita del Congresso Mondiale. La presenza di personalità di indubbia autorevolezza nel mondo dell’ozonote-rapia e la qualità delle relazioni sono state di elevata valenza scientifica. Il 3° Congresso Mondiale di Brescia, è un effettivo e forte punto di riferimento, un confronto delle varie esperienze mondiali, un ulteriore traguardo per l’ozono-terapia, che come è stato confermato è di grande efficacia terapeutica, basso costo ed esiguo rischio nell’ernia del disco e nel dolore muscolo-scheletrico.L’ampia e qualificata partecipazione di vari specialisti in branche mediche e chirurgiche diverse, ma complementari, ha apportato certezze per i medici che si dedicano all’ozonoterapia.

Un grazie per tutto il lavoro che con passione hai svolto.

ANNUNZIATA IZZO E ANTONELLA BERTOLOTTIINTERMED ONLUS

La riuscita del III World Congress of Oxygen-Ozone dell’aprile 2011 a Brescia, è stata eccezionale, soprattutto per la possibilità di confrontarci con i colleghi e di trovare riscontro in alcune scelte che, soprattutto nei Paesi in via di svilup-po, sono a volte difficili. Il nostro punto di vista è stato supportato anche dalla dott.ssa Iabichella,che ci ha suggerito di applicare la TOA anche sull’Ulcera di Buruli. Anche la collega Schwartz Tapia che ha parlato di una patologia che noi trattiamo spesso nei nostri dispensari in Africa, ci ha dato suggerimenti impor-tanti ed utili. Inoltre il trattamento con olio ozonizzato è risultato utilizzabile sulle lesioni che stiamo curando.

JOSIP BURIC

Caro Matteoinnanzi tutto ti volevo ringraziare ancor una volta per avermi dato la possibilità di presentare le mie relazioni Il convegno è stato di alto livello e non lo dico solo per cortesia. Ho incontrato persone di diverse nazionalità il che conferma la internazionalità del convegno e la diffusione del ozono stesso. Nella stragrande maggioranza dei casi del-le presentazioni cui ho avuto modo di assistere devo confessare che non mi aspettavo un livello così elevato di preparazione scientifica ma anche di meto-dologia della ricerca clinica che ha portato alla presentazione stessa. Inoltre, la condotta del intero convegno nella lingua inglese ha dato quel tocco in più al congresso che certamente se lo è meritato. Congratulazioni per l’organizzazio-ne meticolosa e attenta dell’evento. Al prossimo anno.

Sinceri saluti

V. Kumar India

W. KosAustralia

S. Viti PaganelliVenezuela

F. D. UngureanuRomania

J. BuricCroazia

T. YoussefSiria

T. BarkhotkinaUcraina

S. Catelani CardosoBrasile

M. N. MawsoufEgitto

A. Schwartz TapiaSpagna

R. Viebahn-HanslerGermania

A. Martin FranciscoSpagna

J. VyletelkaSlovacchia

B. ClavoSpagna

J. Baeza NociSpagna

R. QuinteroSpagna

M. A. SamyEgitto

R. AlvaradoBolivia

O. A. PepaArgentina

O. BetancourtVenezuela Argentina

H. XiaofungCina

Y. BergeronCanada

H. KonradBrasile

A. StafaAlbania

V. Kumar India

W. KosAustralia

S. Viti PaganelliVenezuela

F. D. UngureanuRomania

J. BuricCroazia

T. YoussefSiria

T. BarkhotkinaUcraina

S. Catelani CardosoBrasile

M. N. MawsoufEgitto

A. Schwartz TapiaSpagna

R. Viebahn-HanslerGermania

A. Martin FranciscoSpagna

L. Cardelli Italia

M. MorettiItalia

G. PellicanòItalia

G. TabaracciItalia

L. ReItalia

A. De MonteItalia

M. LeonardiItalia

C. AndreulaItalia

V. BocciItalia

M. GallucciItalia

A. ZambelloItalia

M. MutoItalia

P. FalzoniItalia

A. CorradiItalia

A. BertolottiItalia

M. SiritoItalia

A. IzzoItalia

R. Dall’AglioItalia

F. ParodiItalia

V. CoviItalia

F. BerniItalia

C. DimauroItalia

M. L. IabichellaItalia

H. GiulianoItalia

L. Cardelli Italia

M. MorettiItalia

G. PellicanòItalia

G. TabaracciItalia

L. ReItalia

A. De MonteItalia

M. LeonardiItalia

C. AndreulaItalia

V. BocciItalia

M. GallucciItalia

A. ZambelloItalia

M. MutoItalia

147

www.centauro.it International Journal of ozone therapy 10:147,2011

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ore9.00:apeRtURa CoRSoregistrazionepartecipanti

ore9.15:dr.Matteobonetti

L’ozono terapia: stato dell’arte.

ore9.45:dr.MarcoMorettiefficacia del trattamento con 02-03 e acido Yaluronico nelle lacerazioni parziali del tendine del sovra spinato.

ore10.15:dr.alessiozambelloautoemoterapia ozonizzata nel trattamento delle ulcere vascolari non responsive al trattamento tradizionale.

ore10.45–11.00:coffeebreak

ore11.00:dr.MarcoMorettiefficacia delle infiltrazioni con ossigeno ozono e acido Yaluronico nella tendinopatia da sovraccarico del tendine d’achille nello sportivo.

ore11.30:dr.alessiozambelloL’ossigeno ozono nella pannicolopatia edematosa degli arti inferiori: trattamento estetico o terapeutico?

ore12.00:dr.MatteobonettiL’ossigeno ozono terapia nel trattamento del mal di schiena.

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The International Journal of Ozone Therapy (IJOT) is a clinical practice journal documenting the current state of ozone therapy practice worldwide. IJOT publishes original clinical observations, descriptions of new techniques or procedures, case reports, and articles on the ethical and social aspects of related health care. Submissions suitable for the Journal include observa-tional studies, clinical trials, epidemiological work, reports on health services and outcomes, and advances in applied (translational) and/or basic research.The instructions for submission of articles to IJOT follow the Uniform Requirements for Manuscripts Submitted to Biomedical Journals of the International Committee of Medical Journal edi-tors (ICMJE, http://www.icmje.org), if not otherwise indicated below.

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Publication Typesoriginal Research articlesarefull-lengthresearchpapers,whicharefavoredbyIJOT.articlescovertopicsrelevanttoclinicalstudies,andmayincludebothbasicandexperimentalwork.Review articlesarecomprehensive,state-of-the-artpapersfocusingonim-portantclinicalproblemsandshouldaddressaspecifictopicinascholarlymanner.sucharticlesmaybe invitedby theeditorormaybeunsolicitedreviews.Case Reports/technical Notes should be unusually educational and medi-cally important.although IJOT preferentially encourages submission offull-lengthoriginalresearcharticles,thejournalwillconsiderpublicationofalimitednumberofconcisecasereportsandtechnicalnotes.editorials areusuallyinvitedbytheeditor.editorial Comments are specific comments on the articles published inIJOTandareusuallyinvitedbytheeditor.Letters to the editorcontainconstructivecommentsorcriticismofaspecificpaperpublishedbyIJOT.Lettersdealingwithsubjectsofgeneral interestwithin the fieldofIJOT,orpersonalopinionsonaspecific subjectwithintheambitofIJOT,mayalsobeaccepted.all papers should be sent to the Scientific editordr.Matteobonetti:[email protected]

Manuscript PreparationIntroductions that Satisfy Reader expectations

– overview: onceyouhavea reviseddraft, youneed toensure thatyourIntroductionframes it, so thatyourreaderswillunderstandwhereyouaretaking them.the Introduction should orient readers and motivate themto read the restof thepaper.the Introductionmustalsomakea contractwiththereaderthataquestionwillbeanswered.– Functions: – toawaken

the reader’s interest. – to be informative enough to prepare readers tounderstand your paper. – Content and organization: 1. common ground➜ context➜ relevant background➜ orients the reader. 2. disruption➜ problem➜ gap in knowledge, Question➜ Motivates the reader. 3.resolution➜response➜promiseofananswer➜Makesacontractwiththereader.1.Common ground:statestheconsensus,sharedunderstanding,commongroundinthefield,what’sknowandnotknown.givesthereadercontextandprovides relevantbackground information,nota literature re-view.2.Disruption: statestheproblem/thequestionthatthepaperaddresses.conveys the significance, i.e. the costof leaving problemunsolved,or thebenefitofsolvingit.3.Resolution: Implicitlypromisesthatyouwillpresentyouranswerintheresultsanddiscussion.trynottostatetheanswer;thathastheeffectofclosingoffthepaperratherthanleadingintoit.When draft-ing:althoughtheIntroductionunfoldsintheaboveorder,1-2-3,whenyou’redrafting,write it ina3-2-1order.bywritingtheIntroduction inthisorder,youaresuretoworkthroughandclearlysetupyourmainpoint,theques-tion that it answers, and to includeonly relevantbackground information.When revising:“thekeyistothinklikeareader--readersexpectandneeda senseof structure.since readers readeachsentence in lightofhowtheyseeitcontributingtothewhole,whenyoureviseitmakessensetodiagnosefirstthelargestelementsofthepaper,thenfocusonthecoherenceofyourparagraphs,theclarityofyoursentences,andonlylastonmattersofspellingandpunctuation.ofcourse,inreality,noonerevisessoneatly;allofusreviseaswego.but,itisusefultokeepinmindthatwhenyourevisefromthetopdown,fromglobalstructuretosectionstoparagraphstosentencestowords,youaremorelikelytodiscoverusefulrevisionsthanifyoustartatthebottomwithwordsandsentencesandworkup”.boothwc,colombgg,williamsjM,etal.thecraftofresearch.chicago:universityofchicagopress;2008.

originalresearchshouldbeorganizedinthecustomaryformat,asdescribedbelow.thetextofthemanuscriptshouldbesubmittedasasingledocumentwiththefollowingsections(inorder):1. author Information page (firstpage) -full titleof thearticle, authors’names,highestacademicdegreeearnedbyeachauthor,authors’affiliations,nameandcompleteaddress forcorrespondence,address forreprints ifdif-ferentfromaddressforcorrespondence,faxnumber,telephonenumber,ande-mailaddress.2. acknowledgments and Funding page (second page) - theacknowledgments section lists all funding sources for the researchof thestudy,anddetailssubstantivecontributionsofindividuals.theauthorsmustrevealallpossibleconflictsofInterest/disclosures.3. title page(thirdpage)-thefulltitle,itemizedlistofthenumberofta-bles,thenumberandtypes(colororblack-and-white)offigures,andthree-to-fivekeywordsforuseasindexingtermsshouldbeincluded.appropriatekeywordsshouldbeselectedfromtheMedicalsubjectheading.thewordcountofthetextshouldbespecified.4. Summary-asummaryofupto250wordsshouldsummarizetheprob-lemspresentedanddescribethestudiesundertaken,resultsandconclusions.sincetheabstractmustbeexplicative,theabbreviationsmustbereducedtoaminimumandexplained.referencesshouldnotbecited intheabstract.5. text-typicalmainheadingsincludeIntroduction,MaterialsandMethods,results,discussion,andconclusions.abbreviationsmustbedefinedatfirstmention in the text, tablesand figures.thecompletenamesand shortad-dressesofmanufacturersofanyequipmentusedinMaterialsandMethodsmustbe supplied. Ifanimalsareused inexperiments, state the species, thestrain,thenumberofanimalsused,andanyotherpertinentdescriptivechar-acteristics.Ifhumansubjectsorpatientsareemployed,provideatablewithrelevantcharacteristics.whendescribingsurgicalorneurointerventionalpro-ceduresonanimals, identify thepre-anestheticandanestheticagentsused,andstatetheamountorconcentrationandtherouteandfrequencyofadmin-istrationofeachagent.genericnamesofdrugsmustbegiven.Manuscriptsthatdescribestudiesonhumansmustindicatethatthestudywasapprovedbyan Institutionalreviewcommitteeand thatall subjectsgave informedconsent.reportsofstudiesonbothanimalsandhumansmustindicatethatall procedures followed were in accordance with institutional guidelines.6. References-citationsshouldbelistedinorderofappearanceinthetext,andbetweensquarebrackets[].referencesmustbelistedattheendofthetextintheorderofcitation.journaltitlesshouldbeabbreviatedaccordingtoIndexMedicus.forcitationrulesnotspecifiedhere,authorsshouldrefertotheNLM Style guide for authors, editors, and publishers (http://www.nlm.nih.gov/citingmedicine).all references must be checked by the author(s).Journal article:Laredojd,bardM.thoracicspine:percutaneous trephinebiopsy.radiology.1986;160:485-489.Journal article if the number of authors is more than six: list the first three authors followed by et al.geibpraserts,kringst,pereirav,etal.clinicalcharacteristicsofduralar-teriovenous shunts in 446 patients of three different ethnicities. Interventneuroradiol.2009;15:395-400.entire book:valavanisa.Medicalradiology:interventionalneuroradiology.heidelberg:springerverlag;1993.part of book if number of authors is more than two:bonnevillejf,clarissej,etal.radiologieinterventionnelle.In:Manelfeced.Imageriedurachisetdelamoelle.paris:vigotediteur;1989.p.761-776.7. tables-eachtablemustbetypedonaseparatesheetanddouble-spaced,ifpossible.tablesshouldbenumberedusingthearabicsystem,followedbyabriefinformativetitle.usetypeofthesamefontandsizeasemployedinthetext.Includefootnotesatthebottomofeachtable.tablesmustbenum-beredintheordercitedinthetext.tablesshouldnotduplicatedatagiveninthetextorfigures.

Rivista di proprietà EDIZIONI DEL CENTAURO S.r.l. - Edita da CENTAURO S.r.l. - Sede legale: Via del Pratello, 8 – I-40122 BolognaC.F. e P. IVA 01896531207 – Cap. soc. Euro 15.600.00 i.v. – Reg. Impr. BO – R.E.A. BO n. 397358

8. Figure Legends - providefigurelegendsonaseparatesheet.Legendsmustbedouble-spaced,andfiguresmustbenumberedintheordercitedinthetext.9. Figures - figures should preferably be submitted online in .tiff formatto:[email protected] timerequired to send fileswill varydepend-ingonthenumberoffigures,butimageresolutionmust not be reducedtodecrease transmission time.when labeling the figures, please ensure thatthelabelcorrespondstothefigurenumber.Digital images (originals or imagesacquired bya scanner) mustmeet thefollowingcriteria:Black-and-white figures:Imagesmustbeacquiredusingthegreyscalewithaminimumresolutionof300pixelsper inchor150pixelspercm.Imagesmusthaveabaseofatleast8.1cmforoneitemoraminimumbaseof16.9cmforseveralitems. Color figures:ImagesmustbeacquiredusingthefullcolorcMykmethodwith a minimum resolution of 300 pixels per inch or 150 pixels per cm.Imagesmusthaveabaseofatleast8.1cmforoneitemoraminimumbaseof16.9cmforseveral items.thergbmethod isrecommendedforvideoreproductionsonly,asthequalityofprintedfiguresispoor.Imagesmustbesavedin.tiff format.Imagesin.jpgformatarenotacceptableasdetailstendtobelostuponscanning,evenathighresolution.Imagedefi-nitionalsodependson theenlargement factor.thus,a large low-resolutionimagecanbeproportionallyreduced(by24%)forpublication,therebyper-mittingoptimalpresentationinprint.however,enlargementofasmallhigh-resolutionimagewillhighlightallflaws,yieldingapixelatedeffect.donotsubmit figuresalreadypaged inword,powerpoint,orotherdocu-ments,orimagesinsertedinwebpages.suchimagesareoflowresolutionandareunsuitableforprinting.figuresin.dcm(dicom)formatmaybesubmittedas.dcmor.tifffiles.theeditorialofficewillprocesssuchimagesforprinting.

IllustrationsmaybecompressedusingthestuffIt,aladdin,orzipprograms.donotlabelanimagewitharrows,numbers,orletters,butratherindicateonaduplicatecopyorasketchwheresuchindicationsaredesired.authorsareadvisedtorefertothejournalguidelineswhenformattingtheirwork.otherwisetheeditorialofficemayreturnasubmissiontotheauthorsforimprovementbeforeanyeditorisassigned.IfitisnotpossibletosendfiguresviatheInternet,imagesmaybesentbyexpresscourierinoneofthefollowingformats:originalx-rayfilms,slides,orglossyoropaqueprints.clearlyindicateonthebackoftheimagethefirstauthor’snameandthenumbercorrespondingtothefigurecaptionandcitationinthetext.figures may be submitted on a cd-roM or dvd, copied in Iso9006format legibleonbothpcandMac.floppydisksarenotrecommended.clearlyindicateononesideofthefigureboththetopofthefigureandthefigurenumber.donotlabeltheactualimagewitharrows,numbers,orlet-ters,butratherindicatethetopandfigurenumberonaduplicatecopyoronasketch.donotcutorattachfigureswithadhesivetapeorusepaperclips.Ifimagesaresentoffline,thumbnailsoftheimagesshouldbeuploadedasasubmissionitemonline,togetherwiththemanuscript,toallowthepublish-ers to reserve space for the original high-resolution images.all materialshouldbesenttotheeditorialoffice:

the International Journal of ozone therapycentauros.r.L.viadelpratello,8I-40122bologna–Italytel:+39.051.227634fax:+39.051.220099e-mail:[email protected]

Manuscriptno.:________________________________________ date:_______________

withrespecttothearticleentitled:_________________________________________________________

theauthorscertifythat:

(Conflicts of Interest/Disclosures)thecorrespondingauthormustdiscloseall relevant financial,personal,and/orprofes-sionalrelationshipswithotherpeopleororganizationsrelatedtothesubjectofthisarticle.Isthereanyactualorpotentialconflictofinterestinthesubjectofthisarticle?

no(___).Ifyes,pleasegivedetailsinthespacebelow.

__________________________________________________________________________________________________________

(Copyright transfer agreement)all rights, title,and interest in themanuscript, includingcopyrightownership,are trans-ferredtocentaurosrlpublisher,bologna.authorsreserveallproprietaryrights(suchaspatentrights)otherthancopyrightandtherighttousepartsofthisarticleintheirownsubsequentworkassoonasthearticleisacceptedforpublication.thearticlemaynotbepublishedelsewherewithoutpermissionfromthepublisher.

(Financial Support) If the authors or their institution(s) is/are affiliated with any organization having a financial interest(includingtheprovisionoffinancialsupport)inthesubjectofthisarticle,theauthorsshouldindicate(a)thenameoftheindustry/institutionprovidingsupportforthestudy,(b)thetypeofaffiliationoftheorganization(stockholder,consultant,donorofhonorarium,grantingbody,sourceofotherfinancialormaterialsupport,providerofequipment,orothersupportcategory),and(c)thattheauthorshadcompletecontrolofthedataandinformationsubmittedforpublication.anyothernon-financialconflictofinterestshouldalsobedisclosedtotheeditor,withtheunderstandingthattheinformationmaybepublishedifdeemedappropriatebytheeditor.

no,I(we)donothaveanyconflictofinterestrelatedtothisarticle(___).

Ifyes,(a)(_________)(b)(_________)(c)(_________).

(exclusive publication Statement)noneofthematerialinthismanuscripthasbeenpublishedpreviouslyorsubmittedforpublicationelsewherepriortoappearanceinnrj.

(author Contribution Form)thecorrespondingauthormustcertify thatallauthorsparticipated inandcontributedsuf-ficientlytotheconceptionanddesignofthework,andanalysisofthedata,aswellasthewritingofthemanuscript,andassume public responsibility for the content of the paper.the corresponding author also certifies that all authors havereviewedthefinalversionofthemanuscriptandapproveditforpublication.

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thissignedstatementmustbereceivedbeforethemanuscriptcanbeprocessedforpublication.

INTERNATIONAL JOURNAL OF OZONE THERAPY Conflicts of Interest/Disclosures, Copyright Transfer Agreement, Financial Support,

Exclusive Publication Statement, and Author Contribution Form

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