ozone therapy and oxidative stress evaluation · ozone therapy - pontevedra. june 5th, 2009. ozone...

Eugenio Luigi Iorio. 2008©

INTERNATIONAL CONFERENCE. New horizons for the ozone therapy - Pontevedra. June 5th, 2009.

Ozone therapy and oxidative stress evaluation

Eugenio Luigi Iorio, MD, PhD

International Observatory of Oxidative Stress (Salerno, Italy)OXI.GEN LAB (Brescia, Italy)


Evolution of living organisms and redox status

Cloroplasti di piante ed alghe verdi

Cloroplasti di alghe rosse Mitocondri

Batteri mobili E. coli Rhizobacteria

Solfo-batteri rossi

Solfo-batteri verdi

EUCARIOTIPROCARIOTI

Atmosfera ossidante

Atmosfera riducenteFotosintesi con H2O

Fotosintesi con H2S

Ciclo di fissazione del C

Cyanobacteria

Batteri rossi non zolfo-dipendenti

Batteri verdi filamentosi

Batteri blu-verdi

Perdita fotosintesi

Respirazione con O2 Respirazione con O2 Respirazione con O2

Batteri fermentanti ancestrali

Perdita fotosintesi


General metabolic pathways

From oxidative processes to energy and live

NAD+ FAD

Nutrienti

Ciclo di Krebbs

Lipidicomplessi

Glicerolo Acidi grassi

Glicidi complessi

Glucosio

Glicolisi

Piruvato

Acetil-CoA

Proteine

Amminoacidi

NH3, CO2, H2O

Prodotti semplici da substrati complessi

Estrazione diequivalenti riducenti

NADH + H+ FADH2

ATP ADP+P O2 H2O

PRODUZIONE DI ENERGIA


Comparison between anaerobiosis and aerobiosis

Much more energy from aerobic oxidative processes

Citoplasma

GLUCOSIO

Rendimento netto

2 ATPGlicolisi

2 piruvato

2 ATP

32 ATP

Mitocondrio

Catena respiratoriaFosforilazione ossidativa

O2

6 CO2

Rendimento netto

+

H2O

Ciclo di Krebbs

2 NADH

2 NADH

6 NADH

2 FADH2


The other face of oxidation


Oxygen and living organisms

The discovery of oxygen toxicity

Prime evidenze sperimentali in animali da laboratorio sul possibile ruolo tossico dell’iperossia1940-1950

Disturbi respiratori fino a comparsa di lesioni alveolari dopo prolungata esposizione ad O2 puro

1954-1955 La fibroplasia retrolenticolare è provocata dall’esposizione dei neonati ad elevata pO2

1960

Molte lesioni in vivo sono provocate dalle specie reattive dell’ossigeno

Alcune specie reattive sono utili (fagocitosi)


The two sides of oxygenThe two sides of oxygen


What is OXIDATIVE STRESS?

The breaking of a balance

The oxidative stress is a particular kind of chemical stress induced by the presence, in a living organism, of an excess of oxidant chemical species (OCS) that results from an increased production of OCS and/or a decreased efficacy of antioxidant enzymatic and/or non enzymatic defence system.

Free radicalsAntioxidantsProtection

from diseases Tissue damage

(ageing and diseases)


Ageing and almost 100 diseases are related to OXIDATIVE STRESS

“The free radical man”

Alzheimer’s disease

Atherosclerosis

Kidney diseases

Pancreatitis and liver diseases

Stroke, miocardial infarction

COPD

Parkinson’s disease

Maculopathy, cataract

Rheumatoid arthritis

Crohn’s disease

Diabetes, obesity

Cancer

AGEING

AGEING


THE INTERNATIONAL OBSERVATORY OF OXIDATIVE STRESS ACTIVITIES

CONTINUOUS EDUCATION

DATA COLLECTING

INFORMATIONRESEARCHINTERNATIONALOBSERVATORY

OXIDATIVE STRESS


AN OXI.GEN LAB VIEW


The REDOXOMICS. A common trait d’union.

GENOMICS

PROTEOMICS

LIPIDOMICS

GLYCOMICS

METABOLOMICSTRASCRIPTOMICS REDOXOMICS


AN OXI.GEN LAB VIEW


FAST TEST®: THE LIPIDOMICS ENTERS IN TO THE POPULATION STUDIES.


From the clinic to the laboratory

Clinical suspect

History and clinical visit

Oxidant/antioxidant status evaluation

↑Chemical oxidant species and/or ↓antioxidants

Generic diagnosis of OXIDATIVE STRESS


Inflammation/infection markers (ESV, CRP, AST, leukocytes)

Abnormal

AbnormalNormalCardiovascular markers(cholesterol, HCY, nitrates?)

Metabolism/respiration markers(BMI, thyroid hormones,LDH,lactate)

Normal

AbnormalNormal

Toxicity/exposure markers (AST, ALT, alcohol, drugs, chemicals)

Abnormal

O. S. by abnormal reactivity

O. S. by metabolic/respiratory deficit

O. S. by pO2 changes

Normal

O. S. by indefinite causes

EMPIRIC APPROACH

EZIOLOGIC DIAGNOSIS

RECOVERY

O. S. MONITORING

SPECIFIC TREATMENT

RELAPSES PREVENTION

O. S. MONITORING

O. S. by microsomial induction

INCREASED OXIDANT STATUS DECREASED ANTOX STATUS

INTAKE/ABSORP/AVAILABILITY

AO ENZ LP/HS AS MINER

FURTHER INVESTIGATIONS

O. S. biochemical assessment


THE PROJECT FOR SPAIN PHARMACISTS


Oxidant Chemical Species (OCS). The Free Radicals.

Free Radicals or Radicals, instead of most atoms, share always one or more unpaired electron(s)

Oxygen Free Radicals (unstable)

The hydroxyl radical (HO.) One unpaired electron

O H

An atom of OxygenTwo unpaired electrons

O

An atom of NeonOnly paired electrons

Ne

Atom (stable)


Non-Radical Oxidant Chemical Species

Non-Radical OCS, like hydrogen peroxide, share always paired electrons

OH O HO H


The REDOX process

The chemical basis of REDOXOMICS

++Free radical

(oxidant)

A A

Neutral molecule (reduced, stable)

Target molecule (e. g. a double bond C-C)

C C

New radical(oxidized, oxidant)

CC

unpaired electronoxidation

H2O2 + HCl → H2O + HClO-1

-1

-2

+1

REDUCTION

OXIDATION

OXIDANT

REDUCING

Gain of electrons

Loss of electrons


Plasma antioxidant barrier prevents cellular oxidative damage by blocking OCS in the blood

Thiol groups (-SH) are qualitatively important components of plasma antioxidant barrier

-SH Other chemical groupsOCS


The production of free radicals from leukocytes

Relevance of oxidative stress

PLA2

PL AA PG

Proteasi

O2O2

.NADP

H-Ox

H2O2

HClO OH. + OH-

SOD

MPx

Cl-

Fe/CuFenton/HW

ROOH

R-NHClR-NH2

Fe/Cu RO• /ROO•

H2O

CAT

/GPx

NOS

Arg

CitNO

NOO-O2

.

KILLING

Batteri, LPS, Ig

Batteri, LPS, Ig

Batteri, LPS, Ig

PLA2

PL AA PG

Proteasi

O2O2

.NADP

H-Ox

H2O2

HClO OH. + OH-

SOD

MPx

Cl-

Fe/CuFenton/HW

ROOH

R-NHClR-NH2

Fe/Cu RO• /ROO•

H2O

CAT

/GPx

NOS

Arg

CitNO

NOO-O2

.

KILLING

Batteri, LPS, Ig

Batteri, LPS, Ig

Batteri, LPS, Ig


Basic cell mechanisms of oxidative tissue damage

External agents

HO•

RH

H2O

R•

O2 ROO •

R1H ROOH

R1•

Oxidative damage

RespirationOxidative damage

Nucleus

Citoplasm

R-O-O-H R-O•

Fe3+Fe2+

R-O-O• R-O-O-H

H+

OH-

pH ↓

LDL oxidation

Endothelial damage

External agents

HO•

RH

H2O

R•

O2 ROO •

R1H ROOH

R1•

Oxidative damage


Nucleus

Citoplasm

R-O-O-H R-O•

Fe3+Fe2+

R-O-O• R-O-O-H

H+

OH-

pH ↓

LDL oxidation

Endothelial damage

PLA2

PL AA PG

Proteases

O2

O2. NADPH-O

xH2O2

HClO OH. + OH-

SOD

MPx

Cl-

Fe/CuFenton/HW

ROOH

R-NHClR-NH2

Bacteria, LPS, Ig Fe/Cu

RO• /ROO •

H2O

CAT

/GPx

Bacteria, LPS, Ig

Bacteria, LPS, Ig

NOS

Arg

CitNO

NOO-O2

.

PLA2

PL AA PG

Proteases

O2

O2. NADPH-O

xH2O2

HClO OH. + OH-

SOD

MPx

Cl-

Fe/CuFenton/HW

ROOH

R-NHClR-NH2


RO• /ROO •

H2O

CAT

/GPx

Bacteria, LPS, Ig

Bacteria, LPS, Ig

NOS

Arg

CitNO

NOO-O2

.+

Water

R HH

Hydrogen radical

H

Hydroxyl radical

RHUV +

Water

R HH

Water

R HH R HH

Hydrogen radical

H

Hydrogen radical

HH

Hydroxyl radical

RH

Hydroxyl radical

RH RHUVUV


Disastrous effects of free radical excess on the cell

Every biomolecule can be affected by OCS

Cellula normale(senza lesioni)

Cellula dopo l’attacco di SCO

Perossidazione di lipidi

Modificazioni enzimatiche

Lesioni del DNA

Rottura di proteine

Perossidazione ammi-noacidi e proteine

(Per)ossidazione di carboidrati

Alterazioni dell’omeostasi ionica


Biochemical basis of atheromatous lesion

The role of dyslipidemia and oxidative stress


The discovery of NITRIC OXIDE as signalling molecule

Together with the Nobel Prize, Louis Ignarro.

NORMAL FUNCTIONS

⇔O2•

NO•

NOSL-arg

ABNORNAL FUNCTIONS

⇑O2•

NO•

NOSL-arg

ONOO-


OXIDATIVE STRESS results by an unbalance between prooxidant and antioxidant systems

Oxidant Chemical species can be either the cause or the effect of OXIDATIVE STRESS

Oxidant Species ↑ Antioxidant defences ↓

Radiations, drugs, heavy metalsCigarette smoking, alcohol, pollutionInadequate exercise, to be sedentary

Infectious and other diseases

Reduced intakeand/or reduced synthesis

and/or reduced bioavailability e/o increased antioxidant consumption

Cell damage

Early aging

Cardiovascular diseases

Other diseases

Alzheimer’s d. Parkinson’s d.

Inflammations,neoplasm

Tissue damage

Organ damageSystemic damage


Oxidative stress. The break of a balance.

Evaluating to prevent and to monitor (OCS can be either the cause or the effect of damage)

Increased production of OCS (↑ROS, RNS, RCS, RAS…)

Antioxidant barrier impairment (IC and/or EC AO ↓)

↑ (Per)oxidation of biomolecules with generation of oxidised byproducts

ROOH, R-NHCl, AGE, IP, 8-OH-dG

↑Oxidised byproducts and/o ↓ concentration/activity of AOinto the tissues and/or extracellular fluids

EARLY AGING AND OXIDATIVE STRESS RELATED DISEASES(stroke, infarction, diabetes, Alzheimer’s and Parkinson’s disesase, cancer)


Biomarker of oxidative stress and relative diseases


The isoprostanes pathway

Overview


The carbonylation pathway

Overview


Other common biomarkers of oxidative stress

Overview

Advanced glycosilation end-products (AGEs)

Chlorine and nitro-derivatives

Markers of DNA oxidative damage

Glutathione-adducts


The challenge of imaging (monoclonal antibodies against 8-OH-dG) (1)

Rat lung slices (atypical dysplasia)


The challenge of imaging (monoclonal antibodies against 8-OH-dG) (2)

Rat lung slices (squammous cystis)


Ozone therapy

.Why and how to monitor oxidative stress

http://www.amazon.com/review/create-review/ref=dp_top_cm_cr_acr_wr_link?ie=UTF8&nodeID=&asin=0934740070


Ozone therapy

.Why and how to monitor oxidative stress?



Ozone therapy monitoring

.Which test?



Ozone therapy

.How to manage the results?



Evaluating to prevent and to monitor

Evaluation of the “attack” Evaluation of the “defenses”

First line tests (d-ROMs, MDA)

First line tests (OXY, TAS, BAP, SHp)

Second line tests(FAT PROFILE®, FAST, IP, 8-OH-dG)

Second line tests(GPx, SOD, CAT, vitamins, minerals)

Global evaluation of oxidative stress

Prevention and monitoring of early aging and oxidative stress-related diseases


The Carratelli’s panel

Evaluating to prevent and to monitor

GLOBAL EVALUATION OF OXIDATIVE STRESS

Evaluation of the “attack” Evaluation of the “defense”

PREVENTING AND MONITORING OF OXIDATIVE STRESS-RELATED DISEASES

d-ROMs test BAP test


d-ROMs test

The chemical principle (normal range 250-300 CARR U)

Serum/plasma blood sample

Buffered acidic solution

DPPD oxidation

Color increasing

Photometric reading Radical cationCOLORED (PINK)

Amine baseNOT COLORED


BAP test

The chemical principle (optimal value>2200 µmol/L)

Serum/plasma blood sample

Thiocianate + FeCl3

Colored complex

Loss of color

Photometric reading


Combined evidence of spectrophotometry and electron spin resonance spectroscopy

The DEPPD radical responsible for the ESR spectrum is also responsible for the absorption at 505 nm

0 25 10050

0

1.0

0.5

Time (min)

EPR

inte

nsity

(arb

itrar

y un

its)

75 0 25 100500

0.8

0.4

Abso

rban

ce a

t 505

nm

(A50

5)

75Time (min)

(A) Room temperature time profile of the normalised spectral intensity (•) and of A505 readings () exhibited by the system DEPPD (3.7 x 10-3 M)/tBuOOH (3.9 x 10-5 M)/FeSO4 (2.8x10-5 M) at room temperature. (B) Time profile of the A505 readinmgs exhibited by the systems DEPPD (3.7 x 10-3 M)/tBuOOH (3.9 x 10-5 M)/FeSO4 (2.8x10-5 M) (•), DEPPD (3.7 x 10-3 M)/tBuOOH (2.0 x 10-5 M)/FeSO4 (2.8x10-5 M) () and DEPPD (3.7 x 10-3 M)/tBuOOH (0.95 x 10-5 M)/FeSO4(2.8x10-5 M) () at room temperature. tBuOOH: tert-buthylhydroperoxide; DEPPD: N,N-diethylparaphenylendiamine


Genetic disorders with brain impairment. The Oxidative Balance Progeria Study.

Significantly increased oxidative status in Progeria (Iorio, 2007)

500

0

100

200

300

400

CA

RR

U

Prooxidant status (d-ROMs)2500

0

500

1000

1500

2000

µmol

es re

duce

d iro

n/L

Antioxidant status (BAP)


The d-ROMs test is a useful test to predict the first atherothrombotic event

Ridke et al. Circulation, 2004



Predictive role of d-ROMs test

Vass

alle

C, B

oni C

, D

i Cec

co P

, La

ndi P

. Ele

vate

d hy

drop

erox

ide

leve

ls a

s a

prog

nost

ic

pred

icto

r of

mor

talit

y in

a c

ohor

t of

pat

ient

s w

ith c

ardi

ovas

cula

r di

seas

e. I

nt J

Car

diol

. 20

05

. Nov

14.

[Ep

ub a

head

of pr

int]

Kaplan–Meier survival curves according to 75th d-ROMs test percentiles, considering cardiac death (A) and total mortality (B) as end points.

B

93%

81%

70

60

50

Surv

ival

rat

e (%

)

70

60

50

Months of follow-up0 5 10 15 20 25

98%

84%

70

60

50

Surv

ival

rat

e (%

)

70

60

50

Months of follow-up0 5 10 15 20 25

A



Predictive role of GPx

Blan

kenb

erg

S, R

uppr

echt

Hj,

Bick

el C

, To

rzew

ski

M,

Haf

ner

G,

Tire

t L,

Sm

ieja

M,

Cam

bien

F,

Mey

er J

, Lac

kner

KJ.

Glu

tath

ione

Per

oxid

ase

1 ac

tivity

and

car

diov

ascu

lar

even

ts in

pat

ient

s w

ith

coro

nary

art

ery

dise

ase.

N E

ngl J

Med

. 20

03.

349

(17

): 1

605

– 16

13.

1.0

0.9

0.8

0.7

0.00.0 100 200 300

Weeks of follow-up

Even

t fr

ee s

urv

ival

Quartile 4 (>56,31 U/g Hb)

Quartile 3(56,31 – 48,32 U/g Hb)

Quartile 2(48,32 – 42,00 U/g Hb)

Quartile 1(> 42,00 U/g Hb)

P


The d-ROMs test

A SIMPLE PROCEDURE FOR SCREENING PURPOSES

5544 66

2211 33

8877 99


THE NEW CARPE DIEM SYSTEM

CARRATELLI’S PANEL EVALUATOR


The model of hyperbaric oxygen therapy (HBO). Suitability of d-ROMs test.

Significant increase of oxidants after repeated cycles of hyperbaric oxygen therapy (Benedetti, 2004)

0

100

200

300

400

500

CA

RR

U

First HBO 15th HBO

Before HBOAfter HBO

Diabetic feet (n = 4) - Refractory chronic osteomyelitis (n = 4) - Aseptic osteonecrosis (n = 4)

p


The model of hyperbaric oxygen therapy (HBO).Suitability of BAP test.

Significant increase of BAP test values after two session of hyperbaric oxygen therapy (n=9, means ± SE, one-way repeated-measures ANOVA followed by Dunnett’s test) (modified by Kongoji et al, 2007).

0

500

1000

2500

µEq/

L

First Session

Before

After

Second Session

1500

2000

P


The model of SCUBA divers.Suitability of BAP test.

Significant increase of BAP test values after diving (n=10, means ± SE, one-way repeated-measures ANOVA followed by Dunnett’s test) (modified by Yamami et al, 2007).

0

500

1000

2500

µEq/

L

Before diving After diving

1500

2000

P


Ozone therapy

TheWimbledon’s Study


Ozone therapy

TheWimbledon’s Study

•Although the trends observed in this limited study suggest the beneficial effect of ozone on the immune system further research on a wider scale especially on the observation of the transient rise in free radical activity 24 hours following ozone application would be welcome.

•As there is the possibility, that in patients suffering from autoimmune conditions, free radical activity may not always fall following this observed 24 hour transient rise.

•One precaution to eliminate this risk would be to prescribe high antioxidant medication for 24 to 48 hours following ozone treatment.


Ozone therapy

.The protective effects of plasma antioxidants



Epithelial cancers

A multi-step process (Go et al.)


Cellular response to oxidation


The paradox of oxidant chemical species (1)


Cancer chemoprevention by ROS?


The paradox of oxidant chemical species (2)

From toxic to therapeutic effect depending on the dose


Chemopreventive –induced oxidative stress may enhance cell elimination


Increased levels of oxidative stress after chemotherapy

Usefulness of d-ROMs test

La T

orre

F, O

rland

o A

, Silip

igni

A, G

iaco

bello

T,

Per

goliz

zi S

, Ara

gona

M M

iner

va M

edic

a. 1

996.

86:

1-4

.

200

250

300

350

400

450

Baseline

CA

RR

U

1th cycle 2th cycle

*

*p


Oxidative stress levels increase during antineoplastic radiotherapy

d-ROMs test is useful to monitor oxidative stress in radio-treated neoplastic patients

La T

orre

F, O

rland

o A

, Silip

igni

A, G

iaco

bello

T,

Per

goliz

zi S

, Ara

gona

M M

iner

va M

edic

a. 1

996.

86:

1-4

.

160

210

260

310

360

410

Baseline

CA

RR

U

Ith cycle 2th cycle

*

** **

*p


Corticotherapy drammatically reduces free radical levels in patients with leukemia/lymphomas

d-ROMs test is useful to monitor immunosuppressive therapy in oncology

Dio

nis

io C

, Po

zzi D

, Li

si A

, San

toro

N,

Set

tim

o G

Rep

ort

of C.N

.R.

After therapyBefore therapy0

100

200

300

400

500C

AR

R U

p


Antioxidant formulas and cancer

Results from a controlled trial


Oxidative stress and cancer

Significant differences between controls and patients



Significant improvement of oxidative stress balance (d-ROMs⇓ and ⇑ GPx) after single supplements



Significant improvement of oxidative stress balance (d-ROMs⇓ and ⇑ GPx) after multiple supplements



Significant improvement of clinical outcomes after antioxidant treatments


Oncology. The SERUM MILK PROTEIN Intervention Study on female breast cancer.

Significant increase of biological antioxidant potential after 12-months supplementation (Landoni, 2009)

200250

300

350400

450500

0[350,29±39,29]

Month 2[379,79±54,24]

Month 4[385,93±71,91]

Month 6[390,93±61,92]

Month 8[380,64±50,22]

Month 10[394,86±57,97]

Month 12[407,00±58,54]

* ** *******

CAR

R U

200250

300

350400

450500

0[350,29±39,29]

Month 2[379,79±54,24]

Month 4[385,93±71,91]

Month 6[390,93±61,92]

Month 8[380,64±50,22]

Month 10[394,86±57,97]

Month 12[407,00±58,54]

* ** *******

CAR

R U

100012001400

16001800

2000

220024002600

0[1894,5±362,5]

Month 2[1909,9±478,2]

Month 4[2259,3±283,0]

Month 6[2244,6±225,9]

Month 8[2361,5±344,5]

Month 10[2267,9±304,0]

Month 12[2319,5±288,7]

Red

uced

iron

(µM

)

100012001400

16001800

2000

220024002600

0[1894,5±362,5]

Month 2[1909,9±478,2]

Month 4[2259,3±283,0]

Month 6[2244,6±225,9]

Month 8[2361,5±344,5]

Month 10[2267,9±304,0]

Month 12[2319,5±288,7]

Red

uced

iron

(µM

)

* ***** **** *****

Statistical comparisons were calculated towards the baseline values (time 0): *p=0,039; **p=0,017; ***p=0,009; ****p=0,002.

Statistical comparisons were calculated towards the baseline values (time 0): *p=0,007; **p=0,009; ***p=0,004; ****p=0,02; *****p=0,007.


Basic cell mechanisms of oxidative tissue damage

External agents

HO•

RH

H2O

R•

O2 ROO •

R1H ROOH

R1•

Oxidative damage


Nucleus

Citoplasm

R-O-O-H R-O•

Fe3+Fe2+

R-O-O• R-O-O-H

H+

OH-

pH ↓

LDL oxidation

Endothelial damage

External agents

HO•

RH

H2O

R•

O2 ROO •

R1H ROOH

R1•

Oxidative damage


Nucleus

Citoplasm

R-O-O-H R-O•

Fe3+Fe2+

R-O-O• R-O-O-H

H+

OH-

pH ↓

LDL oxidation

Endothelial damage

PLA2

PL AA PG

Proteases

O2

O2. NADPH-O

xH2O2

HClO OH. + OH-

SOD

MPx

Cl-

Fe/CuFenton/HW

ROOH

R-NHClR-NH2


RO• /ROO •

H2O

CAT

/GPx

Bacteria, LPS, Ig

Bacteria, LPS, Ig

NOS

Arg

CitNO

NOO-O2

.

PLA2

PL AA PG

Proteases

O2

O2. NADPH-O

xH2O2

HClO OH. + OH-

SOD

MPx

Cl-

Fe/CuFenton/HW

ROOH

R-NHClR-NH2


RO• /ROO •

H2O

CAT

/GPx

Bacteria, LPS, Ig

Bacteria, LPS, Ig

NOS

Arg

CitNO

NOO-O2

.+

Water

R HH

Hydrogen radical

H

Hydroxyl radical

RHUV +

Water

R HH

Water

R HH R HH

Hydrogen radical

H

Hydrogen radical

HH

Hydroxyl radical

RH

Hydroxyl radical

RH RHUVUV


The role of lipoperoxides (LPO) in cell physiology modulation

Girotti, 1998.


OXIDANTS AND CANCER

OUR HYPOTHESIS (CARRATELLI AND IORIO)

LPO ACIDOSIS RO•/ROO• DEATH

[]

Test Test Test

HYPOXIA FREE Fe/Cu


CARRATELLI AND IORIO HYPOTHESIS

STUDIES IN VITRO WITH PEROXIDISED EXTRA-VIRGIN OLIVE OIL


CARRATELLI AND IORIO HYPOTHESIS

STUDIES IN VIVO WITH PEROXIDISED EXTRAVIRGIN OLIVE OIL


Potential usefulness of γ-linolenic acid in cance therapy


Oxidative stress and cancer:a radical approach.


Peroxidative cancer: a possible approach to cancer therapy.


Pro-oxidant action of high ascorbate concentration


INTERNATIONAL CONFERENCE. New horizons for the ozone therapy - Pontevedra. June 5th, 2009.

www.oxidativestressobservatory.orgwww.oxigenlab.com

MUCHAS GRACIAS!

Slide 1Slide 2Slide 3Slide 4Slide 5Slide 6Slide 7Slide 8Slide 9Slide 10Slide 11Slide 12Slide 13Slide 14Slide 15Slide 16Slide 17Slide 18Slide 19Slide 20Slide 21Slide 22Slide 23Slide 24Slide 25Slide 26Slide 27Slide 28Slide 29Slide 30Slide 31Slide 32Slide 33Slide 34Slide 35Slide 36Slide 37Slide 38Slide 39Slide 40Slide 41Slide 42Slide 43Slide 44Slide 45Slide 46Slide 47Slide 48Slide 49Slide 50Slide 51Slide 52Slide 53Slide 54Slide 55Slide 56Slide 57Slide 58Slide 59Slide 60Slide 61Slide 62Slide 63Slide 64Slide 65Slide 66Slide 67Slide 68Slide 69Slide 70Slide 71Slide 72Slide 73Slide 74Slide 75Slide 76Slide 77Slide 78Slide 79Slide 80Slide 81Slide 82Slide 83Slide 84Slide 85Slide 86

ozone therapy and oxidative stress evaluation · ozone therapy - pontevedra. june 5th, 2009. ozone...

Documents