Antioxidants PDF

Copyright DESOFT S.A. 2009 Revista Cubana de Farmacia Vol.
43 Supplement 1 2009, ISSN 0034-7515

Havana Redox 2009
Organized by: University of Havana and Cuban Society of Pharmacology
Sponsorship: International Society for Free Radical Research

Havana Redox 2009 January 29-31, 2009 Hotel Meli Habana, Havana Cuba
Abstract Book Page 1 of 125 Electronic version ISSN 0034-7515

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I II I I In nt te er rn na at ti io on na al l C Co on nf fe er re en nc ce e o of f O Ox xi id da at ti iv ve e S St tr re es ss s
H HA AV VA AN NA A- -R RE ED DO OX X 2 20 00 09 9

Cuban Society of Pharmacology

University of Havana
Pharmacy and Food Science
Colleague

International Society for Free Radical Research
Copyright DESOFT S.A. 2009 Revista Cubana de Farmacia Vol.43 Supplement 1 2009, ISSN 0034-7515

Havana Redox 2009


Index

Scientific committee .............................................................................................................................................................. 5
Organizing committee ........................................................................................................................................................... 5
Conference Agenda .............................................................................................................................................................. 8
Plenary Lecture................................................................................................................................................................... 10
Oxidative Stress and Infectious Disease: From Methodology to Antioxidant Therapeutic Approach ............................ 10
Plant Foods and Beverages: Rich in Antioxidants and Protective for Heart Disease.................................................... 11
Protective Effects of iron Chelation may involve HIF Induction: a Possible Novel Therapy for Parkinsons Disease.... 12
Evaluation of Oxidative Stress in Patients with Chronic Hepatitis C Treated with Pegylated Interferon Associated with
Ribavirin......................................................................................................................................................................... 13
Efficacy of Ozone Oxidative Postconditioning in Patients with Hernia Disc................................................................... 14
Panorama of Science in Cuba ....................................................................................................................................... 15
Redox Environment Puzzle, Pieces Fall in Place with Clinical Diagnostic .................................................................... 16
Accelerated Cytotoxic Mechanism Screening (ACMS) for Idiosyncratic Hepatotoxic Drugs......................................... 17
Free-radical Scavenging and In Vivo Antioxidant Activities of Polyphenol Extracts from Bidens pilosa L. (Asteraceae)
with Anti-inflammatory Properties .................................................................................................................................. 18
Real-Time Monitoring of Oxidants in Disease................................................................................................................ 19
A Procedure to Measure the Antioxidant Properties of Polyphenolic Compounds with DPPHradical assay; Biological
Applications.................................................................................................................................................................... 20
Isolated Mitochondria as Experimental Model for Pharmacological/Toxicological Researches .................................... 21
Exercise and Free Radicals: Harmony and Hormesis? ................................................................................................. 22
Oxidative stress in diabetes, an up-date........................................................................................................................ 23
Antioxidant Usage in Routine Dental Practice ............................................................................................................... 24
A Synergistic Antioxidant Cocktail Attenuates Age and Sucrose-Induced Cardiometabolic Risk by Protecting Meal-
Induced Insulin Sensitization ......................................................................................................................................... 25
Disturbances of Redox Homeostasis in the Response of the Yeast to Environmental Factors and its Modification by
Antioxidants ................................................................................................................................................................... 26
Reactive oxygen species, DNA damage and mutational spectrum. .............................................................................. 27
Nitric oxide, roll in physiology and diseases .................................................................................................................. 28
NO, Peroxynitrite and Matrix Metalloproteinases in Acute and Chronic Heart Failure .................................................. 29
Oxidative Stress and Diseases ........................................................................................................................................... 32
The Impact of Organic Hydroperoxides on Spermatogenesis and Sperm Function...................................................... 32
The Relationship Between the Mutations in the Familial Mediterranean Fever Gene and Blood Oxidative Stress
Markers in Patients with Familial Mediterranean Fever (FMF) ...................................................................................... 33
Oxidative Stress Levels During Familial Mediterranean fever (FMF) Attacks................................................................ 34
Effect of Systemic Colchicine on Oxidative Stress in Patients with Familial Mediterranean Fever ............................... 35
Oxidative Stress in the Blood of Subjects Exposed to a Thermoeletric Industry Emissions (Coal Dust)....................... 36
Benznidazole Therapy Promoted an Oxidative Insult in the Blood of Chronic Chagasic Patients with Cardiopathy..... 38
Oxidative Stress Induction and Antitumoral Effect of Sodium Orthovanadate............................................................... 39
Evaluation of the Oxidative Profile in Ehrlich Ascites Carcinoma in Mice Treated with Cordia Verbenaceae extracts . 40
Evaluation of the Antitumor Mechanism of Casearin-Rich Fraction From Casearia sylvestris Against the Ehrlich Ascites
Carcinoma in Mice ......................................................................................................................................................... 41
Metallothioneins and Zinc Metabolism in the Prevention and Progression of the Diabetes Mellitus and their
Complications: Experimental Evidence, Possible Mechanisms and Clinical Implications. ............................................ 42
Torres A. ........................................................................................................................................................................ 42
Evaluation of the A2 Phospholipase Activity in Hypertensive and Normotensive Patients............................................ 43
Evaluation of the Antioxidant Enzymes Activity in Hypertensive and Normotensive Patients ....................................... 44
Oxidative Damage Markers and DNA Repair Capacity in Two Von Hippel Lindau Familial Cases............................... 45
Sensibility of LDL to Oxidation in the Nefrotic Syndrome. ............................................................................................. 46
Evaluation of Some Oxidative Stress Biomarkers in Pediatric Down Syndrome Patients. ............................................ 47
Oligo and Macroelements Determination on Patients Classified as Normotensive Using Sustained Weight Test........ 48

Havana Redox 2009


Relationship of the Reactive Oxygen Species with Cervicouterine Cancer................................................................... 49
Oxidative Stress and Quality of Life............................................................................................................................... 50
Determination of Oxidative Stress Markers in a Model of Experimental Autoimmune Encephalomyelitis for the Study of
Multiple Sclerosis........................................................................................................................................................... 51
Antioxidant Therapy in Kids Suffering from Congenital Cardiopaties Related to Cyanosis. .......................................... 52
Immunological and Oxidative Stress Mechanisms Involved in the Recombinant Alpha IFN Treatment in Multiple
Sclerosis Patients. ......................................................................................................................................................... 53
Determination of Oxidative Stress Markers in Patients with Cardiovascular Diseases. ................................................ 54
Redox Salival Environment. Comparison Between Patients Affected by Inflammatory Periodontal Disease and
Periodontal Healthy Patients.......................................................................................................................................... 55
Presence of Oxidative Stress in Preclinical Model of Alcoholism. ................................................................................. 56
Determination of the Reactive Oxygen Species in type 2 Diabetic Patients.................................................................. 57
Oxidant Stress in Congenital Heart Diseases. Clinical - Lab Diagnosis. ....................................................................... 58
Oxidative Stress and Its Impact in Cataracts ................................................................................................................ 59
Oxidant Stress in Kids Suffering from Diabetes Mellitus Type 1.................................................................................... 60
T-cell Immunoregulation and Oxidative Stress in Neuromyelitis Optica. ....................................................................... 61
ApoH Polymorphisms, ApoH Plasma Levels and Oxidative Markers in Type 2 Diabetic Patients. ............................... 62
Redox Disruption in Male Infertility ................................................................................................................................ 64
Oxidative stress and pregnancy..................................................................................................................................... 65
Characterization of oxidative stress in spinocerebellar ataxia type 2 patients............................................................... 66
Antioxidants......................................................................................................................................................................... 69
Mitochondrial-Targeted Antioxidants as a Therapy in a Model of Tolerance to Nitroglycerin........................................ 69
Antioxidant Capacity Determination in Citric Juice ........................................................................................................ 70
Total Anthocyanins and Total Phenolics of Capulin (Prunnus capuli) and Their Antioxidant Properties....................... 71
Antioxidant Activity of Loquat (Eriobotrya japonica)....................................................................................................... 72
Neuroprotective Effect of Aqueous Extract of Yellow Maca in Newly Weaned Male Rat .............................................. 73
Antioxidants and Atherosclerosis: Effect of Hydroalcoholic Extracts of Plants for Medical Uses on Key Processes of
the Atherogenesis.......................................................................................................................................................... 74
Vitilvenz

Natural Alternative Therapy in Vitiligo ........................................................................................................... 75
Evaluation of the Antioxidant Potential of Banana (Musa Cavendish, Musaceae) Peel Hydroethanol Crude Extract in a
Wound Healing Model.................................................................................................................................................... 76
Hepatoprotective Effect of 2,4,6-trihydroxiacetophenone Against Oxidative Stress Induced by Carbon Tetrachloride in
Mice ............................................................................................................................................................................... 77
Anthraquinones Extracts From Different Organs of Morinda royoc L. In Vitro Plants and Their Antioxidant Properties.79
Antioxidant and Antigenotoxic Evaluation of DI-31, a Brassinosteroid Analogue.......................................................... 80
Antioxidant Protection of Vascular Endothelium in Oxidative Damage Models to Cultured Endothelial Cells............... 81
Content of Redox-Active Compounds in Foods Consumed in Macrobiotic Diet............................................................ 82
Determination of Antioxidant Capability of Drinks of Jamaica and Tamarind ................................................................ 83
Antioxidant and Wound Healing Effect of Rhizophora mangle (L.) in an Aseptic Wounds Model in Rats. .................... 84
Fruits and Vegetables Ingested by Hypertensive Women: Relationship of The Lipidic Profile and The Antioxidant Total
Capacity in Serum.......................................................................................................................................................... 85
In vitro Antioxidant Activity and Chemical Composition of Halimeda monile Seaweed Aqueous Extract...................... 86
Protective Effect of Bidens pilosa L Extract in Hepatotoxicity Induced by Paracetamol. ............................................... 87
Antioxidant Activity and Atheroprotective Effect of an Aqueous Extract of the Marine Seaweed Halimeda incrassata 88
Musa paradisiacal L Extract (Acitan
): Antioxidant Properties in Inflammatory Models ................................................ 89

Piper auritum HBK Cream: Antinflammatory Activity by Antioxidant Mechanisms. ....................................................... 91
Therapeutic Effect of Mangifera indica L. in Minor Recurrent Aphthous Stomatitis....................................................... 92
Antioxidant Activity of Six Aromatics Plants Used in Cuba as Condiments................................................................... 93
Absorption and Metabolism of Flavonoids and Their Metabolites ................................................................................. 94
Isolation of Antioxidant Compounds from Medicinal Extract of Pedilanthus Tithymaloides........................................... 95
Phenylpropanoid Content in Plant of Theobroma cacao L.: Antioxidant Activity ........................................................... 96

Havana Redox 2009


Mangifera indica L Extract (Vimang) and Its Main Polyphenol Mangiferin Prevent Mitochondrial Oxidative Stress in
Atherosclerosis-Prone Hypercholesterolemic Mouse .................................................................................................... 97
Protective Effects of Mangifera indica L extract (Vimang), and Its Major Component Mangiferin, on Iron-Induced
Oxidative Damage to Rat Serum and Liver ................................................................................................................... 98
Antioxidant Capacity Determination in Mandarin, Maracuya and Grape Juices............................................................ 99
Effect of Aqueous Extract of Yellow Maca in the Levels of Glutathione in the Brain After an Intense Exercise in Newly
Weaned Male Rat ........................................................................................................................................................ 100
Antioxidant Capacity of the Aqueous Extract of Peperomia choroniana D.CD. (ipintaki) by Means of DPPH and
Reception of Hydroxyl Radical..................................................................................................................................... 101
Ferric Reduction Antioxidant Power evaluation on Murraya paniculata L (Jack) tincture. ........................................... 102
Biological activity and chemical composition of Morinda citrifolia liin and Annona muricata extracts.......................... 103
Redox Signaling Mechanisms........................................................................................................................................... 105
Expression, Functionality and Relevance of Sodium-Vitamin C Cotransporter 2 (SVCT2) in Breast Cancer. ............ 105
DHA Reductases and Intracellular Vitamin C Accumulation and Recycling................................................................ 106
Application of NFkappaB Reporter Assays for Detection of Oxidative Challenge in Human Cancer Cell Lines. ........ 107
Role of Cystein Residues in Modulating Ascorbic Acid Transport by SVCT1.............................................................. 108
Regulation of Heat Shock-Induced Apoptosis by Sensitive to Apoptosis Gene Protein.............................................. 109
Effect of Glutathione Depletion by Buthionine Sulfoximine on Oxidative Metabolism and Cell Death in Rat Brain..... 110
Uncoupling and Oxidative Stress in Liver Mitochondria Isolated From Rats With Acute Iron Overload...................... 111
Role of hydrogen peroxide in vitiligo............................................................................................................................ 112
Ozone oxidative preconditioning and postconditioning in acute nephrotoxicity induced by cisplatin in rats................ 113
Biomarkers and Diagnostic Methods ................................................................................................................................ 114
Evaluation of Nitric Oxide Produced by Immune Cells for Determining Biological Activity in Vaccines Containing
Whole-Cell Pertussis.................................................................................................................................................... 114
Easy High Performance FRAP Utilizing Photocolorimetric Microplate (Format 96 Wells)........................................... 116
Optimisation of an EPR Method for Detection of Intracellular Superoxide Radicals in Stimulated Macrophages....... 117
Comet Assay: Application to Evaluate the DNA Repair Capacity in Individuals with Xeroderma Pigmentosum
Presumptive Diagnosis. ............................................................................................................................................... 118
Pharmacological or Dietetic Interventions......................................................................................................................... 119
In Vitro Effect of Trofin
, an Hydrolyzed of Proteins and Heme Iron Mixture on Ferrous Iron Status.......................... 119
Estimation of food ingestion and total antioxidant capacity of the offered diet in two elderly homes in La Habana .... 120
Other ................................................................................................................................................................................. 121
Chenopodium ambrosioides: Antiparasitic Drug that Acts on Mitochondrial Respiratory Chain.................................. 121
Oxidative Stress Associated Parameters in Cytotoxicity Induced by 1-O-alkylglycerols in N2a Cell Line................... 122
Authors Index.................................................................................................................................................................... 123

Editorial and Production Staff: Scientific Committee Havana Redox 2009

Sociedad Cubana de Farmacologa
Revista Cubana de Farmacia
CEDISAP, La Habana, Cuba, January, 2009

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ISSN 0034-7515

Revista Cubana de Farmacia Vol. 43 Supplement 1, 2009

Havana Redox 2009


Honor Presidents
Dr. Jos R. Balaguer Cabrera
Dr. Juan Vela Valds
Dra. Concepcin Campa Huerga
Dr. Oscar Ros Lpez
Scientific committee

President: Gregorio Martnez Snchez, Ph.D.
1

Ren Delgado Hernndez, Ph.D.
2
Olga Sonia Len Fernndez, Ph.D.
1

Tania Bilbao Revoredo, Ph.D.
1

Gilberto Pardo Andreu, Ph.D.
1

Evangelina Marrero, Ph.D.
4

Roberto A Menndez Soto del Valle, Ph.D.
7
Silvia Menendez Cepero Ph.D.
5

Pedram Ghafourifar, PharmD, Ph.D.
6
Organizing committee

President: Ren Delgado Hernndez, Ph.D.
2

Alain Valdivia Acosta, B.Sc.
1

Diadelis Ramrez Figueredo, Ph.D.
5

Gema Prez Davison, Ph.D.
1

Livan Delgado Roche, B.Sc.
1

Luis Alberto Gonzles R de Zrate, B.Sc.
1

Manuel Cedeo Palenzuela, M.Sc.
1

Mara Cristina Lara Bastanzuri, M.D.
3

Mariam Hernndez Colina, M.Sc.
1

Marisol Pea Snchez, M.Sc.
8

Miguel David Fernndez Prez, M.Sc.
7

Nielsen Lagumercindes Dennis, B.Sc.
1

Nubia Fernndez Hernndez, M.Sc.
1

Omar Ramrez Nez, B.Sc.
1

Pedram Ghafourifar, Pharm D, Ph.D.
6

Rodolfo Lpez Prez, M.Sc.
1

Souliem Chang Rodrguez, B.Sc.
1

Yahelin Ferrer, M.Sc.
1
Emilio Acosta Medina, M.Sc.
1

1 Instituto de Farmacia y Alimentos, Universidad de La Habana, Cuba
2 Centro de Coordinador de Ensayos Clnicos, La Habana, Cuba
3 Centro para el desarrollo de la Farmaco-epidemiologa, La Habana Cuba
4 Centro de Sanidad Agropecuaria, La Habana, Cuba
5 Centro de Investigaciones del Ozono, La Habana, Cuba
6 Ohio State University, USA
7 Centro de Investigaciones Marinas, La Habana, Cuba
8 Instituto de Neurologa, La Habana, Cuba


Havana Redox 2009

Sponsors
International Society for Free Radical Research
Univ. Milan. Italy
Univ. Politecnica delle Marche, Ancona. Italy
Ministerio de Salud Pblica
Consejo Nacional de Sociedades Cientficas de la Salud
Sociedad Cubana de Farmacologa
Centro de Qumica Farmacutica
Centro Nacional de Sanidad Agropecuaria
Instituto de Farmacia y Alimentos
Instituto de Vacunas Finlay
Empresa QUIMEFA
LABIOFAM

Others Pharmaceutics Laboratories and Companies
Lab. Alexis Biochemicals
Lab. Rubio-Pharma S.A. (Mxico)
Lab.Jeric S.A. (Venezuela)
Lab. Kybela (Ecuador)


Havana Redox 2009

PREFACE
The Cuban Society of Pharmacology and the Pharmacy & Food Sciences College of the Univertsity
of Havana call its associates and all researchers in the field of free radical / antioxidants in Medicine
and Biology to participate in HAVANA-REDOX 2009. This international meeting will be held in
Havana city from January, 29-31, 2009.

Reactive Oxygen Species have been associated with the aging process, life styles, diet, several
clinical disorders and a range of age-related diseases including cancer, atherosclerosis, and
neurodegenerative diseases such as Parkinsons and Alzheimers diseases. The percentage of the
world population of 65 years of age and older has been growing steadily. As longer life spans
become a reality, there is a new emphasis on diagnostic and therapeutic applications that not only
prolong life, but also increase the quality of life and the productivity of the aging population. Aging is
associated with an increased incidence and prevalence of numerous diseases, many of them
chronic. It is now known that free radicals cause extensive damage to cellular components which
can lead to serious dysfunctions and death. Longevity and preventive medicine will substantially
impact many of the social and economic issues of the 21st century.

Speakers invited by the organizing Committee will advance novel and important concepts and set
new directions in the fields of oxidative stress, redox chemistry and biology. Havana Redox is an
unique opportunity to combine the exciting field of oxidative stress with Cuban treasures, beaches
and peoples. hospitality and friendliness.

Around 100 Cuban researchers from universities, medical colleges, research institutes, hospitals
and pharmacies are going to participate in this meeting, part of the strategies developed worldwide
to encourage the integration and collaboration of professionals as well as their scientific update.
The Organizing Committee highlights the participation of more than 75 scientists from 25 countries.
From America, Europe, Asia, Australia and Africa.

More than 110 scientific contributions among lectures, oral communications and posters
presentations; as well as the debate, analysis and reciprocal exchange will make this congress
especially important. They will also allow the transfer of the acquired knowledge to the participants
institutions in more than 25 countries and will emphasize the need to continue research into
oxidative stress chemistry, normal and abnormal biological functions, oxidative stress-related
mechanisms, methods to measure oxidative stress in humans for clinical purpose,
dietary/pharmacological intervention strategies and the study of antioxidants.

During Havana Redox 2009, our invited speakers will be discussing advanced aspects of the
mechanism and measurements of protein oxidations and their role in aging and diseases; redox
signaling mechanisms, biomarkers of oxidative stress; the role of mitochondrial reactive oxygen
species in cell signaling and diseases; and methods in free radical biology and medicine, and their
application to translational research. We will also have the launching of a book about ozone
therapy.

A great wealth of research will be communicated through a three-day poster symposium and oral
presentation sessions that will be a vital component of the meeting. This is a great opportunity for
young investigators to share their latest results in all aspects of free radical biology and medicine.
Welcome everybody to the wonderful venue of this Congress, Havana City, one of the most
important urban areas of the continent, entitled by UNESCO as Mankind Heritage. We are
convinced that this meeting will bring forth new perspectives for future work and it will consolidate
the scientific activity of all the participants even more.

Thank you for accepting our invitation.
Dr. Gregorio Martnez Snchez, Ph.D.

Havana Redox 2009

Conference Agenda
Wednesday, 28 Room Hierro Hotel Meli Habana
09:00-2:00 Registration
Thursday, 29
09:00-2:00 Registration. Room Hierro. Poster and Exhibitions. Room Palma
09:00-9:30
Welcome / Open ceremony. Room Telde Hotel Meli Habana
Gregorio Martnez Snchez, Ph.D. President Scientific Committee. Welcoming speech: Rene Delgado Hernndez, Ph.D. President of
Cuban Society of Pharmacology
9:30-11:00
Plenary Lecture. Chairs: Ren Delgado, Ph.D. / Dagmar Garca, Ph.D. / Paul Cos Ph.D.
9.30 Paul Cos, Belgium: Oxidative stress and infectious disease: from methodology to antioxidant therapeutic approach
10.00 JA Vinson, USA: Plant Foods and Beverages: Rich in Antioxidants and Protective for Heart Disease
10.30 JK Andersen, USA: Protective effects of iron chelation may involve HIF induction: a possible novel therapy for Parkinsons disease
11:00-11:15 Coffee Break
11:15-12:45
Plenary Lecture. Chairs: Olga Sonia Len, Ph.D. / Lamberto Re, Ph.D. / Richard Schould Ph.D.
11.15 D Wilhem Fiho, Brazil: Evaluation of oxidative stress in patients with chronic hepatitis C treated with pegylated interferon associated with ribavirin
11.45 Olga S Len F, Cuba: Efficacy of Ozone Oxidative Postconditioning in Patients with Hernia Disc
12.15 Silvia Menendez, Cuba: Presentation of the book: Ozone: basics and clinical application
12.45-13-30 Lunch
13.30-14.00
Transfer to Scientific tour West Scientific Community of Havana
14.00 Special lecture: Panorama of Science in Cuba. Eduardo Pentn reas, Ph.D. Center of Genetic Engineering and Biotechnology
16.00- Hotel Transfer
Friday, 30
Room Telde Hotel Meli Habana
9:00-11:00
Plenary Lecture. Chairs: Gregorio Martnez, Ph.D. / J A. Melendez, Ph.D. / Peter OBrien Ph.D.
9.00 Gregorio MS, Cuba: Redox environment puzzle, pieces fall in place with clinical diagnostic
9.30 Peter OBrien, Canada: Accelerated Cytotoxic Mechanism Screening (ACMS) for idiosyncratic hepatotoxic drugs
10.00 RC Pedrosa, Brazil: Free-radical scavenging and in vivo antioxidant activities of polyphenol extracts from Bidens pilosa L.
10.30 J A Melendez, USA: Real-Time Monitoring of Oxidants in Disease
11:15-12:45
Plenary Lecture. Chairs: Gilberto Pardo, Ph.D. / Z Radak, Ph.D. / JK Andersen Ph.D.
11.15 I D Postescu, Romania: A procedure to measure the antioxidant properties of polyphenolic compounds with DPPHradical assay; Biol. Applicati.
11.45 Gilberto P, Cuba: Isolated mitochondria as experimental model in pharmacological-toxicological research
12.15 Z Radak, Hungary: Exercise and Free Radicals: Harmony and Hormesis?
12.45-13-30 Lunch
13.30-16.00 Cultural tour Havana City Tour
16.00- Hotel Transfer
Saturday, 31
9:00-11:00
Plenary Lecture. Chairs: Alberto Nez, Ph.D. / I.D Postescu, Ph.D. / D Wilhem Fiho, Ph.D.
9.00 Alberto N S, Cuba: Oxidative stress in diabetes, an up-date
9.30 Tuncen Ozen, TurKey: Antioxidant usage in routine dental practice
10.00 W W Lautt, Canada: A Synergistic Antiox. Cocktail Attenuates Age and Sucrose-Induced Cardiometabolic Risk
10.30 G Bartosz, Poland: Disturb.of redox homeost. in the resp. of the yeast to environmental factors and its modification by antioxidants
11:15-12:45
Plenary Lecture. Chairs: Tania Bilbao, Ph.D. / WW Lautt, Ph.D. / JA Vinson Ph.D.
11.15 Silvia Daz, Cuba, Reactive oxygen species, DNA damage and mutational spectrum.
11.45 R Delgado, Cuba, Nitric oxide, roll in physiology and diseases
12.15 General discussion. Discussion of prominent Poster section A and B
12.45-13-30 Closing Ceremony
13.30-15.00 Farewell Lunch / Cultural Gala Meli Habana Swimming Pool

Poster A*: Antioxidants / Biomarkers and Diagnostic Methods / Others. Chairs: Roberto Menndez Ph.D, Diadelis Remirez
Ph.D., Rodolfo Lpez M.Sc., G Bartosz Ph.D.
Poster B*: Oxidative Stress and Diseases / Pharmacological or Dietetic Interventions / Redox Signaling Mechanisms.Chairs:
Eva Marrero Ph.D, Miguel David M.Sc., Tatiana Yoldi M.Sc., Tuncen Ozen, Ph.D.
* All poster should be hanging in the morning of Thursday 29 and removed in the afternoon of Saturday 31

Note: If nothing else is specified, all events take place at Hotel Meli- Habana

Havana Redox 2009

Plenary lecture index
Thursday, 29
9.30
Paul Cos, Belgium
Oxidative stress and infectious disease: from methodology to antioxidant therapeutic approach
10.00
JA Vinson, USA
Plant Foods and Beverages: Rich in Antioxidants and Protective for Heart Disease
10.30
JK Andersen, USA
Protective effects of iron chelation may involve HIF induction: a possible novel therapy for
Parkinsons disease
11.15
D Wilhem Fiho, Brazil
Evaluation of oxidative stress in patients with chronic hepatitis C treated with pegylated interferon
associated with ribavirin
11.45
Olga S Len F, Cuba
Efficacy of Ozone Oxidative Postconditioning in Patients with Hernia Disc
14.00
Eduardo Pentn reas, Ph.D. Center of Genetic Engineering and Biotechnology, Cuba
Panorama of Science in Cuba.
Friday, 30
9.00
Gregorio MS, Cuba
Redox environment puzzle, pieces fall in place with clinical diagnostic
9.30
Peter OBrien, Canada
Accelerated Cytotoxic Mechanism Screening (ACMS) for idiosyncratic hepatotoxic drugs
10.00
RC Pedrosa, Brazil
Free-radical scavenging and in vivo antioxidant activities of polyphenol extracts from Bidens pilosa L.
10.30
J A Melendez, USA
Real-Time Monitoring of Oxidants in Disease
11.15
I D Postescu, Romania
A procedure to measure the antioxidant properties of polyphenolic compounds with DPPHradical
assay; Biol. Applicati.
11.45

Gilberto P, Cuba
Isolated mitochondria as experimental model in pharmacological-toxicological research
12.15
Z Radak, Hungary
Exercise and Free Radicals: Harmony and Hormesis?
Saturday, 31
9.00
Alberto N S, Cuba
Oxidative stress in diabetes, an up-date
9.30
Tuncen Ozen, Turkey
Antioxidant usage in routine dental practice
10.00
W W Lautt, Canada
A Synergistic Antiox. Cocktail Attenuates Age and Sucrose-Induced Cardiometabolic Risk
10.30
G Bartosz, Poland
Disturb.of redox homeost. in the resp. of the yeast to environmental factors and its modification by
antioxidants
11.15
Silvia Daz, Cuba
Mutagenic and antimutagenic mechanism mediated by antioxidants.
11.45
R Delgado, Cuba
Nitric oxide, roll in physiology and diseases

Havana Redox 2009

Plenary Lecture

Oxidative Stress and Infectious Disease: From Methodology to Antioxidant
Therapeutic Approach
P. Cos
a
, M. Deschacht
a
, K. Tot
a
, L. Maes
a

a
Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp,
Belgium, paul.cos@ua.ac.be

Oxidative stress is considered as a pivotal factor in the inflammatory processes that are involved in various
infectious diseases. Despite intense research in the last decade, many controversies are still reported in
literature. For example, the role of oxidative stress in AIDS appears to be quite broad and involves alterations
in viral replication, immune function and apoptosis (1). Oxidative stress caused by HIV may even accelerate
the progression to clinical disease. There is also a positive correlation between the virulence of H. pylori
strains and the production of ROS in gastric mucosa. Consequently, antioxidants could play a complementary
role in the treatment of several infectious diseases, but there is still debate about their real therapeutic
potential. First, an important but often neglected prerequisite to the in vivo evaluation of antioxidants is the
actual monitoring of oxidative stress. Current analytical techniques all suffer from specific shortcomings and
profound knowledge of their limitations and pitfalls in addition to stringent validation and activity criteria
becomes essential for the correct evaluation and interpretation of oxidative stress and antioxidant activity (2).
Second, excessive antioxidant protection could lean over the balance from oxidative stress to oxidative
deficit. The latter is deleterious as low levels of ROS do indeed have beneficial biological effects, for example
in phagocytosis. Third, ROS, antioxidants, transcription factors and cytokines are part of a large human
defence network that behaves like a black box.
1. Cos, P. et al. (2004) J. Nat Prod. 67: 284-293. 2. Hermans, N., Cos P. et al. (2007) Curr. Med. Chem. 14: 417-430.

Key: PL1
Type of Presentation: Oral Presentation
Key words: infectious disease, oxidative stress

Havana Redox 2009

Plenary Lecture

Plant Foods and Beverages: Rich in Antioxidants and Protective for Heart
Disease

Joe A. Vinson

Department of Chemistry, Loyola Hall, University of Scranton, Scranton, PA 19510 USA

Recent epidemiological evidence indicates that consumption of plant-based foods and beverages can reduce
the risk of cardiovascular disease. Our research group has measured the total phenolic antioxidants in fruits,
vegetables, beverages, grains, nuts, oils and spices. We have determined US per capita consumption of these
antioxidants using USDA data. Beverages provide the majority of the antioxidants in the US diet. We have
shown that individual phenols found in plant foods and beverages are more powerful antioxidants when
compared to antioxidant vitamins on a molecule to molecule basis using LDL+VLDL oxidation as a heart
disease model. Our research group has used the cholesterol/saturated fat-fed hamster model which in 10-12
weeks produces arterial foam cells, the early sign of atherosclerosis. Citrus extract, grape seed extract, berry
extract, green and black tea, grape juice, red wine, beer, and cocoa powder have been found to significantly
inhibit the hamster atherosclerosis process by both hypolipemic and antioxidant mechanisms. Fat and sugar
produce oxidative stress in the postprandial state and this is a risk factor for a myocardial infarction in humans.
We have demonstrated that phenols in cranberry juice or chocolate significantly inhibit this oxidative stress
and that high fructose corn syrup, fat and sugar are post-prandial pro-oxidants. After subject drank grape
juice, red wine, green or black tea and coffee, these beverages provided post-prandial antioxidant protection
of LDL+VLDL. Greater consumption of plant foods and beverage is suggested as a means to decrease the
risk of heart disease.

Key: PL2
Key words: Plant foods, beverages, Antioxidant, heart disease


Havana Redox 2009

Plenary Lecture
Protective Effects of iron Chelation may involve HIF Induction: a Possible
Novel Therapy for Parkinsons Disease

JK Andersen, D Lee, S Ragapolan, D Kaur

Buck Institute, 8001 Redwood Blvd, Novato, CA 94945 USA

Parkinsons disease (PD) is a neurodegenerative disorder resulting in loss of motor function stemming
primarily from the loss of dopaminergic (DAergic) neurons within the substantia nigra (SN). Several studies
have demonstrated elevated levels of intracellular iron in the Parkinsonian SN but whether this is causatively
involved in dopaminergic SN cell death has been controversial. We previously reported that iron chelation via
expression of the bodys major iron sequestering protein, ferritin, protects against DAergic SN
neurodegeneration associated with the PD-inducing neurotoxin MPTP in young mice
1
. The aim of our current
research is towards exploring whether iron elevation in young animals contributes to PD pathology via its
activation of the enzyme prolyl hydroxylase (PH). When activated, PH can act as an inhibitor of hypoxia-
inducing factor 1 alpha (HIF-1), a transcription factor which activates various genes involved in both
protection against oxidative stress and in regulation of cellular iron metabolism. In our studies, inhibition of
PHD by the specific inhibitor 3,4-dihydroxybenzoate (DHB) in vivo resulted in the stabilization of cytosolic HIF-
1 and significantly protected against MPTP-induced nigral dopaminergic cell loss. MPTP alone results in a
significant increase in striatal iron levels that was found to be attenuated by co-treatment with DHB. In addition
to HIF activation, we observed up-regulation of several downstream HIF-dependent genes including the
mitochondrial antioxidant MnSOD and the iron regulatory protein HO-1 which has recently been demonstrated
to contribute to cellular iron efflux. Additionally, MPTP-induced decreases in the iron export protein ferroportin
were found to revert back to normal in the presence of DHB. In vitro, the HIF pathway was found to be
activated in dopaminergic midbrain-derived rat N27 cells grown at 3% oxygen treated with two PHD inhibitors,
DHB and DOMG, and an iron chelator, SIH. Concordant with in vivo data, MPP
+
elicited an increase in total
intracellular iron that was attenuated in the presence of DHB. In addition, MPP
+
administration resulted in a
concentration-dependent increase in levels of the iron import protein TfR that was significantly reduced in the
presence of DHB. Taken together, data from this study suggest that the protection against MPTP neurotoxicity
as a consequence of iron chelation may be mediated by inhibition of PH and subsequent increases in cellular
HIF-1 levels that in turn activates genes involved in both protection against increased oxidative stress and
dysregulation of cellular iron homeostasis. This study provides novel data extending the therapeutic aspects of
HIF protein to a PD model of neurodegeneration and may prove beneficial in other diseases associated with
metal-induced oxidative stress such as Alzheimers disease and multiple sclerosis.

Acknowledgements: This work was funded by NIH R01 NS041264 (JKA) and a Hillblom fellowship (DL).

References: 1. Kaur et al. (2003) Neuron 37: 1-20.

Key: PL3
Key words: Parkinson, iron chelation, hypoxia inducing factor 1 alpha

Havana Redox 2009

Plenary Lecture

Evaluation of Oxidative Stress in Patients with Chronic Hepatitis C Treated
with Pegylated Interferon Associated with Ribavirin

MS Farias
a
, CM Ribeiro
a
, P Budni
a
, EB Parisotto
a
, EM Dalmarco
a
, RC Pedrosa, D Wilhem Filho
.

a
Departamento Ecologia e Zoologia, Universidade Federal de Santa Catarina, Florianpolis, Brazil. (a) Departamento de Ecologia e
Zoologia, CCB, Trindade, Florianpolis, 88040-900, Brazil. Tel.: +55 48 37216917; fax: +55 48 37215156.

Introduction: The WHO estimates that about 3% of the world population is contaminated by Hepatic C Virus (HCV). HCV leads to
increased production of Reactive Oxygen Species (ROS) and the endogenous antioxidant enzymes together with non-enzymatic
antioxidants are important defenses against ROS overgeneration. Objective: This study aimed to evaluate the oxidative stress in HCV
patients untreated and treated with interferon combined with ribavirin compared to healthy subjects. Methods: Nine untreated HCV
patients (group II), eleven HCV patients treated with interferon combined with ribavirin (group III), and twelve healthy subjects (group I)
were evaluated for biomarkers of oxidative stress in blood. Catalase activity was determined by measuring the decrease in a 10 mM
hydrogen peroxide solution at 240 nm. Superoxide dismutase activity was measured at 480 nm according to the method of epinephrine
autoxidation.

Glutathione reductase activity was measured at 340 nm, by measuring the rate of NADPH oxidation.

Glutathione
peroxidase was assayed at 340 nm, through the glutathione/NADPH/glutathione reductase system, by the dismutation of tert-
butylhydroperoxide.

Glutathione-S-transferase activity was determined at 340 nm using CDNB (1-chloro-2,4-dinitrobenzene) as
substrate and a 0.15 M GSH concentration.

The contents of reduced glutathione (GSH) was measured at 412nm, using the reagent
DTNB, lipid peroxidation in plasma was determined at 535 nm in triplicate by the thiobarbituric acid method (TBARS) and protein
carbonyl (PC) were determined using DPNH. For the evaluation of liver damage, the concentrations of AST, ALT, GGT and ALP were
measured, through a kinetic-colorimetric method using a E: BT 3000 plus, Wiener Lab, 1.7.6.0/RT version equipment. Statistical
comparisons of the different parameters between Group I and II were carried out through independent Student t test, and between
Group II and Group III through paired-simple Student t test, assuming a minimal (two-sided) level of significance of p<0.05. Results:
The results of the analysis of the enzymatic oxidative stress biomarkers showed several alterations among the three groups examined.
The increased activities of SOD (88.3%-p<0.001; 29.4%-p<0.01), CAT (31.2%-p<0.01; 56.0%-p<0.01) and GPx (9.8%; 31.5%-
p<0.001) found respectively in non treated and treated HCV patients when compared to controls, probably reflect the need for an
antioxidant compensation regarding the oxidative injury caused by virus infection and also the combined use of interferon and ribavirin.
Whilst we detected a marked decrease in GR activity in untreated HCV patients compared both to controls (p<0.01). An increase of
145.6% (p<0.01) and 365% (p<0.001) in blood GST activity was detected in treated HCV patients compared to blood donors and non
treated HCV patients, indicating that the combined therapy of interferon and ribavirin upregulated such activity. The concentrations of
TBARS as assessment of lipid peroxidation showed a significant increase (97.2% - p<0.1) in HCV patients compared to controls, whilst
oxidative damage to proteins measured as protein carbonyl levels, a significant increase was observed only in HCV patients treated
with interferon combined with ribavirin compared to controls and non treated HCV patients (76.3%-p<0.001; 75.1%-p<0.001,
respectively). Whole blood GSH concentrations showed significant decreases both in treated and non treated HCV patients compared
to controls (60.5%-p<0.1; 47%-p<0.1, respectively). Regarding the concentrations of liver enzymes, no changes in AST and GGT
concentrations were detected. However, significant increases in alanine aminotransferase (ALT) from non treated HCV patients
compared to controls (311.7%-p<0.001), whilst the concentrations of alkaline phosphatase (ALP) from non treated HCV patients were
lower compared to both blood donors and HCV patients treated with interferon combined with ribavirin (50.3%-p<0.01; 60.5%-p<0.001).
Conclusion: Our results showed that untreated HCV patients and also treated with interferon combined with ribavirin are coping with
oxidative stress, probably as a compensatory mechanism to the increased ROS generation associated with HCV infection and/or the
aforementioned treatment.

Key: PL4
Key words: oxidative stress, hepatitis C, interferon, ribavirin

Havana Redox 2009

Plenary Lecture

Efficacy of Ozone Oxidative Postconditioning in Patients with Hernia Disc

Olga Sonia Len Fernndez

Pharmacology Department /Center for Research and Biological. Evaluation and Food Department of Pharmacy and Food Institute from
University of Havana, Cuba.

It was investigated the redox status of 33 patients with diagnosis of Hernia Disc (HD) by Computarized Axial
Tomography (CAT), Nuclear Magnetic Resonance (NMR) and Clinical Evaluations. Ozone Oxidative
Postconditioning (OzoneOxPost) 20 sessions twice a week, 3-8 mg of ozone in stepped way and increasing
order was used. Treatment was efficacy in 73% of the patients. There was a decrease in the intensity of pain
in 41%. In addition, OzoneOxPost improved the life quality of the patients in essential functions such as For a
time Sitting and Standing Getting up from a low seat, Lifting, Traveling, Walking and Personal Care
according to BACKILL scale. It was demonstrated that 100% of patients showed a severe oxidative stress.
Major changes in the redox parameters were in Superoxide Dismutase (SOD), Total Hydroperoxides (TH),
Advanced Oxidation Protein Products (AOPP), Fructolysine content and Malondialdehyde (MDA) as well as
the relation between them. Taken all together these results provide scientific evidence about the efficacy of
OzoneOxPost in this neurological disorder and suggest that the oxidation of essential proteins involved in the
integrity of the intervertebral disc is one of the primary events that unchain the complex processes associated
to the HD. It is showed a proposal of the event secuences assodiated to HD and the critical points which
represent therapeutical targets of OzoneOxPost. At our knowledge this is the first study which made a
systemic characterization of patients with HD from redox point of view.

Key: PL5
Key words: hernia disc, ozone, postconditioning


Havana Redox 2009

Plenary Lecture
Panorama of Science in Cuba

Eduardo Pentn reas, Ph.D.

Center of Genetic Engineering and Biotechnology. E.Mail: eduardo.penton@cigb.edu.cu

The content of this lecture is not limited to the scope of the congress, but it is intended to provide an overview
of the biomedical and biotechnological research in Cuba. General information is given as a context
background on the country and the impact of life sciences on its development, which may be of interest to
participants. In this sense, the standards of education (almost 100% literacy, primary and secondary studies;
59 university centres) and health (1:165 doctors per inhabitant, 5.3 infant mortality and 77 years of life
expectancy) of the population (11 millions) are emphasized.

Science in Cuba employs >12 000 researchers and professionals (>15% PhD) working in 210 institutions, fully
integrated through collaboration --not competition--, with a GNP expenditure of about 1.9. The Cuban
government has prioritized biotech development in the health care and agricultural systems, creating spin off
centers derived from research and production institutes. Closed Cycle strategies (from research to post-
marketing) are implemented to achieve international competitiveness in quality, cost, novelty, and production
volume. Cuban biotech is supported by its high standards in health and education combined with the
development of biomedical and pharmaceutical products and services for human and agricultural applications.

Research Institutes located in different geographic regions of the country form scientific poles, which are
administrative structures for the coordination, collaboration, complementation and control of research plans
and production capacities of their members. The first and most consolidated one is the West Havana Scientific
Pole comprising 11 institutes with different profiles and complementing potentials. Research and production,
patent development, product sales, strategic alliances, joint ventures, technology transfers, contract
manufacture and scientific collaboration are important activities of these centers at the national and inter-
national levels.

Recombinant proteins (interferon, erythropoietin, streptokinase) and vaccines (hepatitis B, cattle tick), also of
synthetic origin (Hib), monoclonal antibodies (for production, diagnostic and therapeutic use), drugs reducing
cholesterol and free oxygen radicals, are examples of the Cuban biomedical and biotech research output,
benefiting patients of viral infections (HIV, hepatitis), cancer, degenerative and metabolic diseases, both in
Cuba as well as dozens of countries.

Key: PL6
Type of Presentation: Plenary Lecture
Key words: Cuba, research strategies, biomedical research, biotechnological research

Havana Redox 2009

Plenary Lecture

Redox Environment Puzzle, Pieces Fall in Place with Clinical Diagnostic

Gregorio Martnez Snchez

Center for Research and Biological Evaluation (CEIEB), Food and Pharmacy College (IFAL), Universidad de La Habana (UH), Cuba.
E.mail: gregorioms@infomed.sld.cu.

The methodology for detecting oxidative stress status at clinical level is hardly to be found in the literature.
There are some useful methods for investigating the oxidative profile but they are not applicable to the clinical
diagnostic. Worldwide in spite of having a high prevalence of illnesses in which Reactive Oxygen Species are
involved (cancer, diabetes, etc.) an integral diagnostic systems have not been developed. It is also insufficient
the education and preparation of the professional personnel (doctors and related health personnel) to face the
analysis of the data that contributes to a antioxidant/pro-oxidant diagnostic and to impact in the modification of
life habits and other measure directed to correct the redox disruption. In the CEIEB-IFAL-UH it was validate a
group of analytic methods for the diagnosis of the oxidative stress. The validated analytic methodology was
permit to establish the reference values in normal populations, an individualized diagnostic as base to
establish the adequate medical prescription, to fallow the evolution of chronic oxidative disorder and follow the
effect of nutritional or pharmacological intervention. The methodology include clinical marker of biomolecule
damage, antioxidant enzymes, concentration of low level antioxidant and indicators of total antioxidant status.
The analytic methodology was adaptable to micro and ultramicro analytic systems and was validated
according to the international recommendations. Low cost, high precision, fast analysis and integral evaluation
of the redox system in the main characteristic of those methods. The study of an extensive array of oxidative
stress indices permitted the examination of the role of oxidative stress in diabetic patients with
macroangiopatic complications, HIV subjects, patients with neurodegenerative disorders, dengue infected
people, etc. Those results was publishes in international journals. An efficient introduction of those methods in
clinic involves educational programs for physicians or health care personnel.

Key: PL7
Key words: redox status, clinic, diagnosis


Havana Redox 2009

Plenary Lecture

Accelerated Cytotoxic Mechanism Screening (ACMS) for Idiosyncratic
Hepatotoxic Drugs

Peter J. O'Brien

University of Toronto Faculty of Pharmacy Rm 1004, 144 College St. Toronto, Ontario M5S 3M2, Canada peter.obrien@utoronto.ca

One theory for idiosyncratic hepatic drug toxicity is that undetectable hepatic inflammation exposes
hepatocytes to reactive oxygen species, myeloperoxidase and cytokines released by immune / inflammatory
cells e.g. Kupffer cells or infiltrated neutrophils. This has been shown with in vivo rat studies. We have
developed a hepatocyte inflammation in vitro model for screening potential idiosyncratic drugs . A marked
increase (x 3-80 fold) in idiosyncratic drug toxicity was found when hepatocytes were exposed to drugs and
non cytotoxic H
2
O
2
levels and peroxidase. ACMS studies showed mitochondrial oxidative stress was the
molecular cytotoxic mechanism for hepatocyte death induced by the following drugs:- troglitazone,
amodiaquine, hydralazine, isoniazid. Other drug cytotoxicities e.g. ibuprofen, aspirin were not affected by H
2
O
2
with or without MPO. This H
2
O
2
inflammation model should prove to be a more robust screen for assessing the
idiosyncratic hepatotoxicity potential of drugs or drug candidates.

Key: PL8
Key words: hepatotoxicity, idiosyncratic, screening, hydrogen peroxide, H
2
O
2


Havana Redox 2009

Plenary Lecture
Free-radical Scavenging and In Vivo Antioxidant Activities of Polyphenol
Extracts from Bidens pilosa L. (Asteraceae) with Anti-inflammatory
Properties

M.R Kviecinski
a
, KB Felipe
a
, EA Ferreira
a
, MH Rossi
b
, R Geremias
c
, D Wilhelm Filho
a
, RC Pedrosa
a
.

a
Depto. de Bioqumica,
b
Centro de Sanidade Animal do Instituto Biolgico de So Paulo, Universidade Federal de Santa Catarina,
Cidade Universitria, CEP 88040-900, Florianpolis, Brazil.
c
Universidade do Extremo Sul Catarinense, Av. Universitria, CEP 88806-
000, Cricima, Brazil

The Bidens pilosa .L. (Asteraceae) a traditional plant used in Brazilian medicine as anti-inflammatory was
screened for in vitro and in vivo antioxidant activities versus anti-inflammatory property. The plant extracts
inhibited the H2O2/FeCl3 induced oxidation of deoxyribose, in particular the ethyl acetate (EtOAc) extract
(IC50=4.3 4.5 g/ml). High level of scavenger activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical was
detected in the ethanol extract (EtOH) (IC
50
=32.3 6.7 g/ml). In addition, it was demonstrated that this
fraction showed a strong decrease of lipoperoxidation in hepatocytes (IC50=25.1 + 4.5 g/ml). In accordance
with these data, the total soluble phenol constituents showed greater concentrations in EtOAc extracts (3.6
0.5 gallic acid equivalents-GAE/mg). The in vivo antioxidant activity assay with EtOH and EtOAc extracts
showed protection against hepatic oxidation of lipids (28% decrease of lipoperoxidation), protein carbonylation
(>80% of protection) and DNA fragmentation (>50% of protection) associated with significant increase of ferric
reducing\antioxidant power in plasma after treatment with a stressor agent (CCl
4
). Interestingly, data obtained
in acute inflammation model in vivo (Carrageenan paw edema assay) indicated that the EtOAc extract had
also an anti-inflammatory activity (55% of reduction in the paw edema). The results suggest that the anti-
inflammatory effect of this extract could be explained, at least in part, by it antioxidant properties.

Key: PL9
Key words: Bidens pilosa, antioxidant, anti-inflammatory


Havana Redox 2009

Plenary Lecture
Real-Time Monitoring of Oxidants in Disease

Pauline M. Carrico
a
, Nadine Hempel
b
, Amanda Melillo
b
, Jaya Dasgupta
b
, Stewart Sell
a
, J. Andres Melendez
b

a
Ordway Research Institute,
b
Albany Medical College, Albany, NY , USA

In recent years the activity of reactive oxygen and nitrogen species (ROS/RNS) has extended beyond their
role in damaging biomolecules to that of efficient signal transducing molecules that regulate cellular fate and
function. During the course of our work we have identified a number of signaling networks that are particularly
sensitive to subtle shifts in the steady-state production of ROS/RNS that participate in diverse processes
related to tumor progression & metastasis, aging and host resistance to infectious pathogens. A primary hurdle
related to the study of redox-signaling is the ability to monitor oxidant production in cellular signaling
compartments in real-time. Our collaborative team has recently developed a redox-sensing GFP (roGFP)
molecule to monitor oxidant production at sites of active signaling in real-time. Here we present data indicating
that roGFP constructs can be used to monitor oxidant production in metastatic tumor cells, detect shifts in
oxidant production as cells senesce and can monitor the ability of bacterial antioxidants to restrict host-cell
oxidant production. The roGFP probe can help in defining the precise contribution of oxidants in regulating cell
fate and function by allowing for the precise kinetic visualization of ROS/RNS production in real time.

Key: PL10
Key words: redox signal, disease, oxidative stress.

Havana Redox 2009

Plenary Lecture
A Procedure to Measure the Antioxidant Properties of Polyphenolic
Compounds with DPPHradical assay; Biological Applications

Ion-Dan Postescu
a
,Tiberius Dicu
b
, Gabriela Chereches
a
, Adriana Filip
c
, C.Tatomir
a
, M. Perde- Schrepler
a

a
Oncologic Institute Prof.dr. I. Chiricuta, Department of Radiobiology and Tumor Biology, 34-36 Republicii str., 400015, Cluj-Napoca,
Romania
b
University Babes-Bolyai, Faculty of Environmental Sciences, Cluj-Napoca, Romania
c
University of Medicine and Pharmacy I. Hatieganu, Faculty of Pharmacy, Cluj-Napoca, Romania.

Plant-derived antioxidants, due to their phenolic compounds content are reported as potential
candidates for reducing the levels of oxidative stress in living organisms. Therefore, the evaluation of
the antioxidant properties, particularly of dietary plant polyphenols, becomes of major importance and
there are numerous methods serving this purpose. In the present study we measured the antioxidant
activities of 5 synthetic phenols (gallic acid, tannic acid, caffeic acid, hydroquinone and quercetin)
and 4 natural plant-extracts (Vitis vinifera variety Burgund Mare, Calluna vulgaris, Camellia sinensis,
Epilobium hirsutum), by stable free radical DPPH

assay using a new computation procedure. EC
50

(substrate concentrations to lower the initial DPPH

activity by 50%) were calculated by non-linear
regression analysis (sigmoidal four parameters curve) at different intervals of time. Steady state
efficient concentration (EC
50ss
) and the T
EC50ss
, time required to reach steady state, were obtained
from an exponential decay equation. Redox reaction stoichiometry n, was calculated, as well.
Antioxidant capacity varied in order: tannic acid >quercetin gallic acid >hydroquinone> caffeic acid
for pure compounds, respectively: C.sinensis > V.vinifera (BM) >E.hirsutum> C. vulgaris, for plant
extracts. T
EC50ss
values

had different ranking, hydroquinone having the fastest kinetic followed by
caffeic acid, galic acid, tannic acid and quercetin. In the series of natural products the reaction kinetic
decreased as follows: V. vinifera (BM) > C. vulgaris > E.hirsutum > C.sinensis. The results are in
good agreement with data reported in literature. The procedure we described is fast, reproducible
and the parameters might be useful indices in characterizing the radical-scavenging properties of
antioxidants and to choose the doses in biologic experiments carried out with natural extract
samples. For instance, from EC
50
value of V. vinifera (BM) grapes seed extract, we found that this
preparate, at doses bellow its own cytotoxic threshold, decreased doxorubicin induced lipid
peroxidation, in normal fibroblasts (Hfl1) to values below basal levels, in a linear dose-dependent
manner (p< 0.01 by Anova; p< 0.001 by Dunnett). On contrary, in Mls tumor cells the effect was
reversed, the doxorubicin oxidative effect increasing with V. vinifera (BM) concentrations in a dose-
effect relationship (p<0.001 by Anova; p< 0.05 by Dunnett

Key: PL11
Key words: antioxidant polyphenols, plant extracts, DPPH assay, antioxidant activity, doxorubicin

Havana Redox 2009

Plenary Lecture
Isolated Mitochondria as Experimental Model for
Pharmacological/Toxicological Researches

Gilberto L Pardo Andreu

Centro de Estudios para las Investigaciones y Evaluaciones Biolgicas. Instituto de Farmacia y Alimentos .Universidad de La Habana.
Ave. 23 # 21425 e/ 214 y 222, La Coronela, La Lisa. CP 13600, Ciudad Habana, Cuba.

Mitochondrial dysfunction is inherent in a variety of human disorders from the classical mitochondrial
diseases arising from mitochondrial DNA mutations (encephalomyopathies) to those involving
mitochondrial signaling pathways to the rest of the cell, modulated by organellar dynamics and
culminating in programmed cell death. Beyond minding the cells energy status, mitochondria are
major cellular sources (and targets) of free radicals, play key roles in the regulation of calcium
homoestasis, and are effectors of the intrinsic apoptotic pathway due to the release of signaling
molecules that activate and trigger specific caspase cascades. The electron-transfer protein
cytochrome c has been recognized over 20 years ago as a signaling molecule released from
mitochondria, thus changing the view of these organelles from their energy-transducing functions to a
regulatory role in cell death pathways. Hence, it is not surprising that the use of mitochondria as
pharmacological or toxicological experimental model together with the development of mitochondrion-
targeted therapeutics have become primary focus of scientific research. In this regard, our laboratory
has used isolated mitochondria to assess the antioxidant ability of different compounds measuring
the oxygen consumption and the H2O2 production in antimycin A inhibited respiration condition.
Damage at the level of mitochondria have been reported after ischemia and reperfusion of organs.
We have used an in vitro model of isolated-mitochondria anoxia-reoxygenation to investigate the
specific damage (and protection) at the level of mitochondrial oxidative phosphorylation. We have
also utilized the state 3 and 4 respirations, uncoupled and inhibited respiration and ADP/O ratio to
characterize the effects of acute iron overload at mitochondrial level. We have also extensively
studied the effect of polyphenolic compounds on mitochondrial permeability transition (MPT), looking
for anticancer or neuro/cardio-protective potential drugs. MPT is a process involved in apoptotic or
necrotic cells death and it is currently identified as pharmacological and toxicological target. Finally,
we do believe that in vitro studies with mitochondria are of fundamental value since the complexity of
the in vivo system prevents from understanding and describing precise mechanisms and locations of
mitochondrial impairment. These studies cannot simply be transposed to in vivo situation and should
be interpreted with caution.

Key: PL12
Key words: mitochondria, experimental model, pharmacology, toxicology


Havana Redox 2009

Plenary Lecture
Exercise and Free Radicals: Harmony and Hormesis?

Z. Radk

Faculty of Physical Education and Sport Science, Semmelweis University, 1123, Alkotas u. 44, Budapest, Hungary.

The hormesis theory purports that biological systems respond with a bell-shaped curve to exposure to
chemicals, toxins, and radiation. Here I extend the hormesis theory to include reactive oxygen species (ROS).
It is further suggested that the beneficial effects of regular exercise are partly based on the ROS generating
capability of exercise, which is in the stimulation range of ROS production. Indeed, physical inactivity leads to
increased incidence of a variety of diseases and it can be regarded as one of the end points of the exercise-
associated hormesis curve. On the other hand, regular exercise, with moderate intensity and duration, has a
wide range of beneficial effects on the body including the facts that it improves cardio-vascular function, partly
by a nitric oxide mediated adaptation, and may reduce the incidence of Alzheimers disease by enhanced
concentration of neurotrophins and by the modulation of redox homeostasis. Mechanical damage-mediated
adaptation results in increased muscle mass and increased resistance to stressors. Physical inactivity or
strenuous exercise bouts increase the risk of infection, while moderate exercise up-regulates the immune
system. Single bouts of exercise increase, and regular exercise decreases the oxidative challenge to the body,
whereas excessive exercise and overtraining lead to damaging oxidative stress and thus are an indication of
the other end point of the hormetic response. Based upon the genetic setup, regular moderate physical
exercise/activity provides systemic beneficial effects, including improved physiological function, decreased
incidence of disease and a higher quality of life.

Key: PL13
Key words: exercise, free radical, hormesis


Havana Redox 2009

Plenary Lecture

Oxidative stress in diabetes, an up-date

Alberto J. Nez Sells

Centro de Qumica Farmacutica, Apdo. 16042, CP 11600 Habana, Cuba. E.Mail: nunez500412@homail.com

El conocimiento sobre la relacin entre el estrs oxidativo (EO) y la progresin de la diabetes mellitus (DM) ha
avanzado de forma considerable en la ltima dcada. El incremento de los productos de la peroxidacin
lipdica y la oxidacin de aminocidos y protenas (glicoxidacin) a medida que progresa la DM es un claro
indicador de dicha relacin. Dentro de la diversidad de hiptesis que tratan de explicar el surgimiento de las
complicaciones vasculares, disfunciones del endotelio y neuropatas, la hiptesis del EO parece tener grados
de sobrelapamiento con la mayor parte de ellas. La terapia anti-oxidante de la DM cobra cada da mayor
atencin de la comunidad mdica, en la medida que se conocen los resultados de las investigaciones sobre
los fundamentos bioqumicos del EO y sus marcadores en fluidos biolgicos. Sin embargo, existe an la
necesidad de desarrollar y demostrar la utilidad de marcadores bioqumicos especficos del EO en su relacin
con la progresin de la DM, sobre todo en las etapas tempranas de la enfermedad. Mas an, se necesita
conocer hasta qu punto el EO puede ser causa (terapia profilctica) o consecuencia de la progresin de la
DM y sus complicaciones. El presente trabajo discute los resultados publicados recientemente sobre dichos
aspectos y destaca la necesidad de continuar las investigaciones sobre la terapia anti-oxidante de la DM,
desde la etiopatognesis de la enfermedad hasta el desarrollo de ensayos clnicos con un diseo adecuado,
para dar respuesta a las preguntas que an quedan por responder sobre un aspecto de la salud humana que
afecta a ms de 150 millones de habitantes de este planeta.

Key: PL14
Key words: diabetes mellitus, oxidative stress, antioxidant therapy


Havana Redox 2009

Plenary Lecture

Antioxidant Usage in Routine Dental Practice

Tuncer OZEN

Glhane Military Medical Academy, Dental Science Centre, Oral Diagnosis
and Radiology Department, Ankara,TURKEY

Recent years have shown the role of antioxidant supplementation in prevention and/or management of
numerous dental disorders, such as periodontitis and peri-implant disease. Dietary and other enzymatic
antioxidants protect the lipids of lipoproteins and other biomembranes against oxidative damage by
intercepting oxidants before they can attack the tissues. In addition, antioxidant macronutrituents may prevent
increased cytokine production, which is a result of prolonged activation of the immune response. Thus, an
adequate antioxidant intake from both diet and supplementation can be a valuable adjunct in the treatment of
chronic inflammatory dental disorders. A wide array of antioxidants has potential applications in dental
therapeutics and most of them are easily available as dietary supplements. Although antioxidant therapy could
have a promising role in dental practice, a few unaddressed issues need to be clarified before it is firmly
established. For example, high dose supplements of some vitamins may pose a health risk. Another important
problem is drug interactions. In addition, administration of antioxidants may interfere with essential defense
mechanisms. In conclusion, and evidence based approach is needed before making recommendations about
safety and standardization of antioxidant usage in routine dental practice.

Key: PL15
Key words: antioxidant, dental practice


Havana Redox 2009

Plenary Lecture

A Synergistic Antioxidant Cocktail Attenuates Age and Sucrose-Induced
Cardiometabolic Risk by Protecting Meal-Induced Insulin Sensitization

W.W. Lautt

Department of Pharmacology & Therapeutics, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada, R3E 0T6.

Meal-induced insulin sensitization (MIS) is demonstrated by the dramatic increase in insulin response that
occurs following a meal. MIS occurs as a result of a selective increase in response of the skeletal muscle.
Absence of meal-induced insulin sensitization (AMIS) leads to postprandial hyperglycemia, compensatory
hyperinsulinemia and resultant hyperlipidemia and increased oxidative stress. AMIS increases progressively
with age. MIS is 119% in young rats decreasing to 20% after one year of normal aging. The degree of AMIS is
dramatically potentiated by administration of a low dose sucrose supplement but the synergistic antioxidant
cocktail, consisting of S-adenosylmethionine, vitamin E, and vitamin C, protects against the development of
AMIS so that after one year on the sucrose diet untreated rats have MIS of 7% whereas the treated group has
MIS of 75%. Using these methods of increasing and decreasing MIS, we were able to describe an AMIS
syndrome that represents the earliest metabolic defect in the prediabetic state that leads to a progressive
predictable series of dysfunctions, including generalized adiposity. This study demonstrates the concept of an
AMIS syndrome and shows that specific targeting of the MIS process offers dramatic protection against the
development of prediabetic insulin resistance and the associated progression of dysfunctions. Further, if a full
state of AMIS is allowed to develop as a result of a high sucrose supplemented diet, MIS can be restored to
normal levels by targeting the signaling pathway that regulates the MIS process. In order for a meal to lead to
MIS, two feeding signals must be delivered to the liver, the first being a parasympathetic reflex and the second
being an elevation of hepatic glutathione levels. Neither signal alone is sufficient to allow MIS to occur whereas
mimicking both the parasympathetic signal (bethanechol) and the elevated hepatic GSH signal (n-
acetylcysteine) results in complete restoration of MIS. These novel findings strongly support the concept of
targeting the AMIS syndrome through the use of both a preventative, to inhibit the development of AMIS, and a
therapeutic, to reverse AMIS once it has developed.

References: 1. Lautt, W.W. (2004) J Pharmacol Sci 95:9-17. 2. Lautt, W.W. et al. (2008) Exp Gerontol 43:790-800.

Key: PL16
Key words: Meal-induced insulin sensitization, antioxidant, bethanechol


Havana Redox 2009

Plenary Lecture
Disturbances of Redox Homeostasis in the Response of the Yeast to
Environmental Factors and its Modification by Antioxidants

Grzegorz Bartosz
a,b
, Tomasz Bilinski
a

a
Department of Biochemistry and Cell Biology, University of Rzeszow and
b
Department of Molecular Biophysics, University of Lodz,
Poland.

Environmental agents cause alterations in the redox equilibrium of the yeast Saccharomyces cerevisiae. We
have employed yeast mutants deficient in various elements of the antioxidant barrier to study the importance
of antioxidant proteins to these factors and protective effects of antioxidants. Disruption of CuZn-superoxide
dismutase (SOD) brings about auxotrophy to lysine and methionine, increased sensitivity to superoxide-
generating agents and hydrogen peroxide and also to salt stress, heat shock, many metals and xenobiotics
including aldehydes. Exogenous antioxidants are protective in all these cases. Thus, antioxidant-deficient
yeast yeast can be a sensor of both oxidative stress-inducing agents and antioxidants.

Replicative and chronologic aging of yeast cells also induce changes in the redox balance due to decreases in
the activities of antioxidant enzymes and in the contents of antioxidants. Both replicative and chronologic
lifespan is decreased in CuZn-SOD disruptants and can be extended be antioxidant supplements. However,
there is a considerable selectivity in the ability of exogenous antioxidants to protect the yeast against oxidative
stress, many antioxidants efficient in vitro being ineffective at the cellular level. Enhancement of endogenous
antioxidant defence protects against numerous environmental insults. Therefore, overexpression of enzymes
directly involved in antioxidant defence or involved in the biosynthesis of low-molecular weight antioxidants
may enhance the resistance of the yeast to environmental stressors and enhance its usefulness for
biotechnology.

Key: PL17
Key words: Saccharomyces cerevisiae, antioxidant enzyme


Havana Redox 2009

Plenary Lecture

Reactive oxygen species, DNA damage and mutational spectrum.

Silvia Daz Llera

Department of Pharmacology and Toxicology, Pharmacy and Food Sciences College, 25
th
Ave. 21425, 11600, Havana, Cuba.
sdllera@infomed.sld.cu.

Reactive oxygen species (ROS) such as the superoxide ion and hydroxyl radicals are formed continuously by
intracellular metabolism and as a result of environmental exposures. ROS induced damage to cellular
macromolecules has been implicated in the etiology of cancer and ageing, as well as several other human
diseases. Hydrogen peroxide (H
2
O
2
), an endogenous product of normal cell metabolism, can cause a variety
of damage to genetic material in intact cells including DNAprotein cross-links, strand breaks and oxidation
products at sugars or bases. Base damage includes modifications such as thymine glycol,
formamidopyridines, and 8-oxyguanine. The kind and severity of DNA damage vary considerably and may
depend on cell type, as well as culture and treatment conditions. H2O2 has been shown to cause mutations in
bacteria and fungi, and to be clastogenic in mammalian cells in vitro, but reports on its ability to induce gene
mutation in mammalian cells have been inconsistent. Reactive oxygen species (ROS) produced by
intracellular metabolism are believed to contribute to spontaneous mutagenesis in somatic cells. H2O2 has
been shown to induce a variety of genetic alterations. The kinds of DNA sequence alterations caused by H2O2
in prokaryotic cells have been studied extensively, whereas relatively little is known about the mutational
spectrum induced by H
2
O
2
in mammalian genes. We have used the T-cell cloning assay to study the ability of
H
2
O
2
to induce mutations at the hypoxanthine guanine phosphoribosyltransferase (HPRT) locus in primary
human lymphocytes. Treatment of cells for 1 h with 0.341.35 mM of H2O2 caused a dose dependent
decrease of cell survival and increase of the HPRT mutant frequency (MF). After 8 days of expression time,
the highest dose of H
2
O
2
caused a 5-fold increase of MF compared to the untreated control cells. Mutant
clones were collected and the genomic rearrangements at the T-cell receptor (TCR) -locus were studied to
identify independent mutations. RT-PCR and DNA sequencing was used to identify mutations in the HPRT
coding region. Due to a relatively high frequency of sibling clones, only six independent mutations were
obtained among the controls, and 20 among the H2O2 treated cells. In both sets, single base pair substitutions
were the most common type of mutation (5/6 and 13/20, respectively), with a predominance of transitions at
GC base pairs, which is also the most common type of HPRT mutation in T-cells in vivo. Among the single
base pair substitutions, five were new mutations not previously reported in the human HPRT mutation
database. Overall, the kinds of mutation occurring in T-cells in vivo and H
2
O
2
treated cells were similar, albeit
the number of mutants was too small to allow a meaningful statistical comparison. These results demonstrate
that H2O2 is mutagenic to primary human T-lymphocytes in vitro and induces mutations of the same kind that is
observed in the background spectrum of HPRT mutation in T-cells in vivo.

Key: PL18
Key words:


Havana Redox 2009

Plenary Lecture

Nitric oxide, roll in physiology and diseases

Ren Delgado Hernndez

Centro Coordinador de Ensayos Clnicos, La Habana, Cuba. E.Mail: rdelgado@infomed.sld.cu

In contrast with the short research history of the enzymatic synthesis of nitric oxide (NO), the introduction of
nitrate-containing compounds for medicinal purposes marked its 150th anniversary in 1997. Glyceryl trinitrate
(nitroglycerin) is the first compound of this category. On October 12, 1998, the Nobel Assembly awarded the
Nobel Prize in Medicine or Physiology to scientists Robert Furchgott, Louis Ignarro, and Ferid Murad for their
discoveries concerning NO as a signaling molecule in the cardiovascular system. NO-mediated signaling is a
recognized component in various physiologic processes (eg, smooth muscle relaxation, inhibition of platelet
and leukocyte aggregation, attenuation of vascular smooth muscle cell proliferation, neurotransmission, and
immune defense), to name only a few. NO has also been implicated in the pathology of many inflammatory
diseases, including arthritis, myocarditis, colitis, and nephritis and a large number of pathologic conditions
such as amyotrophic lateral sclerosis, cancer, diabetes, and neurodegenerative diseases. Some of these
processes (eg, smooth muscle relaxation, platelet aggregation, and neurotransmission) require only a brief
production of NO at low nanomolar concentrations and are dependent on the recruitment of cyclic guanosine
monophosphate (cGMP)-dependent signaling. Other processes are associated with direct interaction of NO or
reactive nitrogen species derived from it with target proteins and requires a more sustained production of NO
at higher concentrations but do not involve the cGMP pathway.

Key: PL19
Key words: NO, nitric oxide

Havana Redox 2009

Plenary Lecture

NO, Peroxynitrite and Matrix Metalloproteinases in Acute and Chronic Heart
Failure

Richard Schulz

Cardiovascular Research Group, University of Alberta, Edmonton, Alberta, Canada richard.schulz@ualberta.ca

Enhanced generation of NO, superoxide and peroxynitrite are common to several heart diseases including
ischemia-reperfusion (I/R) injury and inflammatory heart disease resulting from exposure to pro-inflammatory
cytokines. My lab wishes to understand how peroxynitrite causes acute contractile dysfunction of the heart.
Matrix metalloproteinases (MMPs) are activated and their endogenous inhibitors, the TIMPs, are inactivated
by peroxynitrite. Although best known for their ability to cleave extracellular matrix proteins, we found that
MMP-2 is localized to discrete intracellular sites including the sarcomere, nucleus and caveolae of cardiac
myocytes. Infusion of peroxynitrite, I/R injury, or cytokine exposure of the heart results in MMP-2 activation and
proteolysis of susceptible intracellular targets in the sarcomere and cytoskeleton including troponin I, myosin
light chain-1 and alpha-actinin, causing contractile failure. Other targets of MMP-2 inside the cell are likely and
being sought. Recent evidence from our lab shows that MMP-2 interaction with caveolin-1 or its
phosphorylation at serine and/or threonine residues inhibit its activity. A paradigm has emerged that MMP-2 is
a key effector of acute oxidative stress injury in the heart, in particular to peroxynitrite, by proteolysis of specific
intracellular targets. Although MMP inhibitor development focused on the extracellular form of the enzyme
activated by proteolytic processing, it is of key importance to develop inhibitors which target the full-length,
peroxynitrite-activated form of MMP-2. MMP inhibitors, including the tetracycline antibiotics such as
doxycycline which possess MMP inhibitor activity, are a viable means to treat/prevent cardiac diseases
associated with enhanced oxidative stress.

Key: PL20
Key words: NO, peroxynitrite, metalloproteinases, heart failure

Havana Redox 2009

Oxidative Stress and Diseases

Index

Oxidative Stress and Diseases ..............................................................................................Error! Marcador no definido.
The Impact of Organic Hydroperoxides on Spermatogenesis and Sperm Function.........Error! Marcador no definido.
The Relationship Between the Mutations in the Familial Mediterranean Fever Gene and Blood Oxidative Stress
Markers in Patients with Familial Mediterranean Fever (FMF) .........................................Error! Marcador no definido.
Oxidative Stress Levels During Familial Mediterranean fever (FMF) Attacks...................Error! Marcador no definido.
Effect of Systemic Colchicine on Oxidative Stress in Patients with Familial Mediterranean Fever ..Error! Marcador no
definido.
Oxidative Stress in the Blood of Subjects Exposed to a Thermoeletric Industry Emissions (Coal Dust)................ Error!
Marcador no definido.
Benznidazole Therapy Promoted an Oxidative Insult in the Blood of Chronic Chagasic Patients with Cardiopathy
..........................................................................................................................................Error! Marcador no definido.
Oxidative Stress Induction and Antitumoral Effect of Sodium Orthovanadate..................Error! Marcador no definido.
Evaluation of the Oxidative Profile in Ehrlich Ascites Carcinoma in Mice Treated with Cordia Verbenaceae extracts
Evaluation of the Antitumor Mechanism of Casearin-Rich Fraction From Casearia sylvestris Against the Ehrlich Ascites
Carcinoma in Mice ............................................................................................................Error! Marcador no definido.
Metallothioneins and Zinc Metabolism in the Prevention and Progression of the Diabetes Mellitus and their
Complications: Experimental Evidence, Possible Mechanisms and Clinical Implications.Error! Marcador no definido.
Torres A. ...........................................................................................................................Error! Marcador no definido.
Evaluation of the A2 Phospholipase Activity in Hypertensive and Normotensive Patients...............Error! Marcador no
definido.
Evaluation of the Antioxidant Enzymes Activity in Hypertensive and Normotensive Patients ..........Error! Marcador no
definido.
Oxidative Damage Markers and DNA Repair Capacity in Two Von Hippel Lindau Familial Cases..Error! Marcador no
definido.
Sensibility of LDL to Oxidation in the Nefrotic Syndrome. ................................................Error! Marcador no definido.
Evaluation of Some Oxidative Stress Biomarkers in Pediatric Down Syndrome Patients. ...............Error! Marcador no
definido.
Oligo and Macroelements Determination on Patients Classified as Normotensive Using Sustained Weight Test. Error!
Relationship of the Reactive Oxygen Species with Cervicouterine Cancer......................Error! Marcador no definido.
Oxidative Stress and Quality of Life..................................................................................Error! Marcador no definido.
Determination of Oxidative Stress Markers in a Model of Experimental Autoimmune Encephalomyelitis for the Study of
Multiple Sclerosis..............................................................................................................Error! Marcador no definido.
Antioxidant Therapy in Kids Suffering from Congenital Cardiopaties Related to Cyanosis. .............Error! Marcador no
definido.
Immunological and Oxidative Stress Mechanisms Involved in the Recombinant Alpha IFN Treatment in Multiple
Sclerosis Patients. ............................................................................................................Error! Marcador no definido.
Determination of Oxidative Stress Markers in Patients with Cardiovascular Diseases. ...Error! Marcador no definido.
Redox Salival Environment. Comparison Between Patients Affected by Inflammatory Periodontal Disease and
Periodontal Healthy Patients.............................................................................................Error! Marcador no definido.
Presence of Oxidative Stress in Preclinical Model of Alcoholism. ....................................Error! Marcador no definido.
Determination of the Reactive Oxygen Species in type 2 Diabetic Patients.....................Error! Marcador no definido.
Oxidant Stress in Congenital Heart Diseases. Clinical - Lab Diagnosis. ..........................Error! Marcador no definido.

Havana Redox 2009

Oxidative Stress and Its Impact in Cataracts ...................................................................Error! Marcador no definido.
Oxidant Stress in Kids Suffering from Diabetes Mellitus Type 1.......................................Error! Marcador no definido.
T-cell Immunoregulation and Oxidative Stress in Neuromyelitis Optica. ..........................Error! Marcador no definido.
ApoH Polymorphisms, ApoH Plasma Levels and Oxidative Markers in Type 2 Diabetic Patients. ..Error! Marcador no
definido.
Redox Disruption in Male Infertility ...................................................................................Error! Marcador no definido.
Oxidative stress and pregnancy........................................................................................Error! Marcador no definido.
Characterization of oxidative stress in spinocerebellar ataxia type 2 patients..................Error! Marcador no definido.


Havana Redox 2009

The Impact of Organic Hydroperoxides on Spermatogenesis and Sperm
Function

YG Aboua
a
, SS du Plessis
b
, N Brooks
a

a
Faculty of Health and Wellness Sciences, Department of Biomedical Sciences, CPUT, Cape Town,
b
Division of Medical Physiology,
Faculty of Health Sciences, University of Stellenbosch, Tygerberg, South Africa

Decreasing trends in male fertility in terms of sperm count and sperm quality have been correlated to
exposure to environmental contaminants in both wildlife and humans. Pollutants such as
hydroperoxides (HP) used in the chemical and pharmaceutical industries and by-products of fuel oil
slicks may cause abnormal spermatogenesis and testicular malfunctioning in the male. In this study,
we have investigated the susceptibility of sperm in the testes and epididymis of rat to hydroperoxide-
induced oxidative stress (OS) in vivo. Male Wistar rats aged 10-12 weeks were placed randomly in
three groups and fed ad libitum standard rat chow (SRC) and water. Solutions of t-butyl
hydroperoxide (tbHP) [5; 10; 20; 40 M], cumene hydroperoxide (cHP) [2.5; 5; 10; 20M] or saline
were administered peritoneally on a daily basis for 5 consecutive days a week for 8 weeks.
Epididymal sperm and testicular supernatants were used to analyze sperm function and enzymes.
The testes and epididymis were also processed for histological assessment. Paired Students t-test
and ANOVA were used to analyze results. No distinctive clinical signs of cytotoxicity and mortality
were found in any of the treatment groups. However, tbHP (20 and 40 M) and cHP (10 and 20M)
significantly decreased superoxide dismutase (SOD), GSH-Glo Glutathione assay activities as well
as sperm cells motility and viability. All these results strongly point to an increase in OS. We can
conclude that we have successfully created an animal model to test the adverse effects of OS on
male reproductive parameters, which will also allow us to study possible treatments in vivo.

Key: OSD1
Type of Presentation: Poster
Key words: hydroperoxides, sperm toxicity, animal model

Havana Redox 2009

The Relationship Between the Mutations in the Familial Mediterranean
Fever Gene and Blood Oxidative Stress Markers in Patients with Familial
Mediterranean Fever (FMF)

C. Oktenli
a
, S. Celik
a
, Y. Tunca
b
, M.H. Terekeci
a
, O. Ipcioglu
c
, Z. Yesilova
d
, S. Nalbant
a

a
GATA Haydarpasa Training Hostipal, Division of Internal Medicine (a), Tibbiye Cad., TR-34668 and Istanbul, Turkey.
b
Gulhane
Military Medical Academy, Department of Medical Genetics (b) , Etlik, TR-06018 and Ankara, Turkey.
c
GATA Haydarpasa Training
Hostipal, Division of Biochemistry (c), Tibbiye Cad., TR-34668 and Istanbul, Turkey.
d
Gulhane Military Medical Academy, Department
of Gastroenteroloji (d) , Etlik, TR-06018 and Ankara, Turkey.

Familial Mediterranean fever (FMF) is an autosomal recessively inherited autoinflammatory disorder
characterized by self-limited attacks of fever and recurrent serozal inflammation episodes. Mutations in the
Familial Mediterranean Fever Gene (MEFV) cause the FMF. The gene MEFV, which is located on human
chromosome 16p and encodes a protein named pyrin. Pyrin is expressed primarily in neutrophils, eosinophils,
cytokine-activated monocytes, and serosal and synovial fibroblasts. Based on the nature of FMF attacks, it has
been speculated that wild-type pyrin acts as a regulator of the inflammatory response, though the precise
nature of this regulatory activity remains to be identified. In a more recent study, it has been shown that pyrin
modulates the apoptotic response to oxidative stress mediated by Siva, a fifth pyrin-interacting protein.
Although several studies have indicated oxidative system abnormalities in patients with familial Mediterranean
fever, it is still obscure whether mutation type in MEFV gene has an effect on the oxidative system in FMF.
Twenty-seven male patients with FMF were enrolled into the study. Patients were investigated for the
presence of five pyrin gene mutations (M694V, M680I, V726A, R761H and M694I), encountered commonly in
our population. When we compared the all parameters between patients with the M694V homozygote
mutations (n = 13) and patients with any other heterozygote mutations (n = 14), serum lipid hydroperoxide
levels were significantly higher (6.33 0.94 nmol vs. 3.64 0.79 nmol; p < 0.0001), and serum superoxide
dismutase (0.09 0.008 U/ml vs. 0.14 0.02 U/ml; p < 0.0001) and catalase enzyme activities (2.06 0.52
nmol/min/ml vs. 4.53 0.94 nmol/min/ml; p < 0.0001) were significantly lower in patients with the M694V
homozygote mutations than the other genotypes. In conclusion, we suggested that patients with homozygote
M694V have higher oxidant stress and lower antioxidant enzyme concentrations than those of other
genotypes.Our findings support the notion that homozygote M694V mutation is associated with a more severe
disease course. Finally, this study has, for the first time, examined the relationship between mutations in the
MEFV gene and oxidative stress in FMF and offers new insights into the underlying mechanisms in the
disease.

Key: OSD2
Key words: Familial Mediterranean fever, oxidative stress, serum

Havana Redox 2009

Oxidative Stress Levels During Familial Mediterranean fever (FMF) Attacks

Z. Yesilova
a
, M.H. Terekeci
b
, O. Yildiz
c
, S. Celik
b
, O. Ipcioglu
d
, S. Nalbant
b
, C. Oktenli
b

a
Gulhane Military Medical Academy, Department of Gastroenteroloji (a) , Etlik, TR-06018 and Ankara, Turkey.
b
GATA Haydarpasa
Training Hostipal, Division of Internal Medicine (b), Tibbiye Cad., TR-34668 and Istanbul, Turkey.
c
Gulhane Military Medical Academy,
Department of Medical Pharmacology (c) , Etlik, TR-06018 and Ankara, Turkey.
d
GATA Haydarpasa Training Hostipal, Division of
Biochemistry (d), Tibbiye Cad., TR-34668 and Istanbul, Turkey.

It is well known that, increased reactive oxygen species (ROS) overwhelms the protective systems and results
in cell damage and lipid peroxidation. Some of the lipid peroxidation products can trigger diverse aspects of
cellular responses, including pro-inflammatory and anti-inflammatory responses. This may play a crucial and
perhaps causative role in the pathogenesis of some inflammatory diseases. Familial Mediterranean fever
(FMF) is an autosomal recessive autoinflammatory disorder predominantly affecting Turks, Sephardic Jews,
Levantine Arabs, and Druze. FMF begins in childhood with recurrent attacks of pain and fever accompanied by
synovitis, peritonitis, and pleuritis. As FMF is a chronic autoinflammatory disease, there may be an abnormality
in the oxidative stress level of FMF patients. During attacks, there are usually neutrophilia and production of a
brisk acute-phase protein, and substantial influx of polymorphonuclear leukocytes into the affected tissues. It
has been suggested that the persistent respiratory burst caused by activated neutrophils may generate ROS
including free radicals that can cause oxidative damage. However, little is known about oxidative system
during attacks in FMF patients. Fifty-five male patients with FMF and 25 body mass index- and age-matched
healthy male volunteers were recruited into the study. All patients were evaluated during attack-free period. In
addition, fifteen of the 55 patients gave an additional blood sample during an attack period (double donors).
Attack-free period (defined as being free of attacks for at least 3 weeks) and acute phases were determined
based on clinical (fever, abdominal pain, arthritis, e.g.) and laboratory findings (high levels of fibrinogen, white
blood cell, and erythrocyte sedimentation rate e.g.). The mean levels of serum lipid hydroperoxide were higher
in both patients with FMF than healthy controls (4.62 1.36 nmol vs. 1.78 0.45 nmol; p < 0.0001) and
patients on acute attack than those of attack-free period (6.26 0.89 nmol vs. 3.54 0.86 nmol; p < 0.0001).
FMF patients have lower serum superoxide dismutase (SOD) (0.11 0.02 U/ml vs. 0.22 0.03 U/ml; p <
0.0001) and catalase (CAT) (3.15 1.49 nmol/min/ml vs. 24.47 5.81 nmol/min/ml; p < 0.0001) enzyme
activities than healthy controls. Patients in acute attack period have also lower serum SOD (0.09 0.008 U/ml
vs. 0.14 0.02 U/ml; p < 0.0001) and CAT (2.04 0.49 nmol/min/ml vs. 4.34 1.16 nmol/min/ml; p < 0.0001)
enzyme activities than those of attack-free period. In conclusion, the principal findings of the present study are
a rise in serum levels of lipid hydroperoxide and depletion of antioxidants such as serum SOD and CAT
enzyme activities during attack period. This may be a finding related with effects of acute inflammation on
oxidative system.

Key: OSD3
Key words: familiar Mediterranean fever, oxidative stress, inflammation

Havana Redox 2009

Effect of Systemic Colchicine on Oxidative Stress in Patients with Familial
Mediterranean Fever

O. Yildiz
a
, S. Celik
c
, O. Ipcioglu
d
, Z. Yesilova
d
, M.H. Terekeci
b
, S. Nalbant
b
, O. Sayan
e
, C. Oktenli
b

a
Gulhane Military Medical Academy, Department of Medical Pharmacology (a) , Etlik, TR-06018 and Ankara, Turkey.
b
GATA
Haydarpasa Training Hostipal, Division of Internal Medicine (b), Tibbiye Cad., TR-34668 and Istanbul, Turkey.
c
GATA Haydarpasa
Training Hostipal, Division of Biochemistry (c), Tibbiye Cad., TR-34668 and Istanbul, Turkey.
d
Gulhane Military Medical Academy,
Department of Gastroenteroloji (d) , Etlik, TR-06018 and Ankara, Turkey.
e
GATA Haydarpasa Training Hostipal, Division of Hematology
(e), Tibbiye Cad., TR-34668 and Istanbul, Turkey.

Familial Mediterranean fever (FMF) is the most common of the inherited autoinflammatory disorders and is
characterized by recurrent, self-limited episodes of fever and serositis. Colchicine is used chiefly in the
treatment of FMF, and is effective in preventing the attacks. Although the precise mechanism of colchicine in
FMF is unknown, colchicine has been demonstrated to suppress the release of fibronectin and has anti-
inflammatory effects, which may be attributed to inhibition of polymorphonuclear leukocyte metabolism,
chemotaxis, or release of cytokines. There is evidence indicating an increased oxidative stress in FMF patients
both in the remission period and in the attack period. However, there have been little information about effect
of colchicine on oxidative stress parameters in patients with FMF. In a more recent report, colchicine
suppresses neutrophil superoxide production in a murine model of gouty arthritis. It has been also speculated
that the early effect of colchicine may be related to antioxidant properties, including inhibition of superoxide
derivate formation. Fifty-five male patients with FMF were included into the study. All patients with FMF, except
15 newly diagnosed, were taking colchicine. Patients taking colchicine (n = 40) have lower serum lipid
hydroperoxide (3.50 0.76 nmol vs. 5.76 1.20 nmol; p < 0.0001), and higher serum catalase (4.44 1.09
nmol/min/ml vs. 2.23 0.87 nmol/min/ml; p < 0.0001) and superoxide dismutase (0.14 0.02 U/ml vs. 0.09
0.01 U/ml; p < 0.0001) enzyme activities than those of non-colchicine users (n = 15). In conclusion, patients
taking colchicine have lower serum lipid hydroperoxide levels and higher antioxidant levels than those of non-
colchicine users. In this regard, systemic colchicine use may be considered as prophylactic therapy, via
decreased oxidative stress, beginning from younger age aimed at lowering some undesirable risk factors. This
study also has, for the first time, investigated the effects of colchicine on oxidative stress parameters in FMF
and offers new insights into the mechanisms by which colchicine may be of benefit.

Key: OSD4
Key words: familiar Mediterranean fever, oxidative stress, colchicine

Havana Redox 2009

Oxidative Stress in the Blood of Subjects Exposed to a Thermoeletric
Industry Emissions (Coal Dust)

FP Possamai
a,b
, S vila Jnior
a,b
, EB Parisotto
a
, AM Moratelli
a
,DB Incio
a
,TR Garlet
a
, P Budni
a
, F
Dal-Pizzol
b
, RC Pedrosa, D Wilhelm Filho
a

a
Depto. de Ecologia e Zoologia, Universidade Federal de Santa Catarina, Cidade Universitria, CEP 88040-900, Florianpolis, Brazil.
b
Universidade do Extremo Sul Catarinense, Av. Universitria, CEP 88806-000, Cricima, Brazil

Introduction: Mineral coal is the most important energetic source in world, Brazil presenting 0,3%t. Brazilian
coal can be found mainly in south and southeast regions where it is used for energy generation. In the process
of energetic generation, emissions of coal dust expose the surrounding population to increased Reactive
Oxygen Species (ROS) generation mainly via the heavy metal contamination. Therefore, it is important to
monitor the antioxidant status in subjects exposed to coal dust because such airborne contamination is a
serious threat to human health. Objectives: The purpose of the present study is to measure the antioxidant
status of workers from a thermoelectric plant, before and after the supplementation with vitamin C (500mg)
and E (800mg), compared to non-exposed subjects (blood donors). Methods: Four groups were studied:
residents (people living ca. 2.5km away) pre and post-suplementation (Rpre, Rpost), workers indirectly
exposed (administration staff) pre and post suplementation (WIpre, WIpost), workers directly exposed to coal
dust, pre and post suplementation (WDpre, WDpost) and also a control group (C, blood donors). Biomarkers of
oxidative stress such as levels of TBARS and protein carbonyls in plasma; levels of whole blood reduced
glutathione (GSH); erythrocytic activities of glutathione S-transferase (GST) and catalase ( CAT) and blood
contents of Protein-thiols were measured spectrophotometrically, and the results were expressed as mean
values standard errors. Differences between multiple groups were compared using ANOVA and the Dunnett
post-hoc test, using a minimal significance of p<0.05. Results: As shown below, TBARS and protein carbonyl
levels were significantly increased whilst all the antioxidant enzymes activities were induced in residents and
workers compared to controls. Antioxidant supplementation was able to attenuate lipoperoxidation and protein
damage and to normalize the activities of antioxidant enzymes when compared to controls. TBARS in plasma
(C: 61.5914.67; Rpre: 112.0913.21; Rpost: 84.2711.23; WIpre: 133.4718.97; WIpost: 84.9512.86;
WDpre: 138.7013.57; WDpost: 83.789 nmoles/mL). Protein carbonyls (C: 1.940.24; Rpre: 3.840.36;
Rpost: 1.220.19; WIpre: 5.650.86; WIpost: 1.010.23; WDpre: 4.350.59; WDpost: 1.070.15 nmol.mg
-
1
protein). GSH content (C: 0.670.09; Rpre: 10.1; Rpost: 0.660.06; WIpre: 0.920.05; WIpost: 0.720.05;
WDpre: 0.940.1; WDpost: 0.620.04 moles/mL). Erythrocytic activities of GST (C: 13.472.63; Rpre:
28.536.28; Rpost: 17.172.95; WIpre: 49.879.55; WIpost: 9.631.53; WDpre: 43.177.29; WDpost:
9.081.17 moles/min/mL). CAT (C: 34.464.53; Rpre: 37.595.62; Rpost: 37.514.28; WIpre: 78.515.4;
WIpost: 42.724.09; WDpre: 80.726.31; WDpost: 41.424.11 mmoles/min/mL). Protein thiols (C:
219.6020.24; Rpre: 177.4718.37; Rpost: 401.5643.30; WIpre: 148.3212.32; WIpost: 462.2046.14;
WDpre: 128.9217.19; WDpost: 308.7619.75 nmol.mg
-1
protein). Conclusions: An oxidative stress condition
were found in the blood of subjects either living near the thermoelectric plant or directly or indirectly exposed to
environmental contamination derived from coal combustion. After antioxidant supplementation, a protective
effect of vitamins C and E on the antioxidant system was evident.

Key: OSD5

Havana Redox 2009

Key words: coal dust, blood oxidative stress

Havana Redox 2009

Benznidazole Therapy Promoted an Oxidative Insult in the Blood of Chronic
Chagasic Patients with Cardiopathy

CM Ribeiro
a
, P Budni
a
, MS Farias
a
, EB Parisotto
a
, RC Pedrosa
b
, RC Pedrosa
a
, D Wilhelm Filho
a
,

a
Departamento Ecologia e Zoologia, CCB, Trindade, 88040-900. Tel.: +55 48 37216917; fax: +55 48 37215156. Universidade Federal
de Santa Catarina, Florianpolis, Brazil. (a)
b
Hospital Clementino Fraga Filho, Servio de Cardiologia - Rua Prof Rodolpho Paulo
Rocco n 255. Cidade Universitaria-Ilha do Fundo- Rio de Janeiro, 21941-913, Brazil. Tel: +55 21 25989494.

Introduction: Chagas' disease is one of the most important public health problems in Latin America with strong incidence in Brazil .
Benznidazole (BZN) is the only drug used for the treatment of chagasic patients in Brazil. However, BZN generates reactive oxygen
species (ROS) through the biotransformation process of xenobiotics. The interaction of both its reactive metabolites and ROS
generation that damage important biomolecules such as DNA, proteins and lipids, are responsible for its chemotherapeutic actions
against the parasite and toxic side effects in the host. Objective: The purpose of the present study was evaluate the biomarkers of
oxidative stress after BZN treatment in patients with Chagas heart disease. Methods: Forty one patients divided in four groups with
Chagas heart disease were selected and biomarkers of oxidative stress were measured before and after two months of BZN treatment
(5mg/kg/day). The four groups were classified according to the modified Los Andes clinical hemodynamic classification in groups IA
(n=10), IB (n=20), II (n=7) and III (n=4). Catalase activity was determined by measuring the decrease in a 10 mM hydrogen peroxide
solution at 240 nm. Superoxide dismutase activity was measured at 480 nm according to the method of epinephrine autoxidation.

Glutathione reductase activity was measured at 340 nm, by measuring the rate of NADPH oxidation.

Glutathione peroxidase was
assayed at 340 nm, through the glutathione/NADPH/glutathione reductase system, by the dismutation of tert-butylhydroperoxide.

Glutathione-S-transferase activity was determined at 340 nm using CDNB (1-chloro-2,4-dinitrobenzene) as substrate and a 0.15 M
GSH concentration.

The contents of reduced glutathione (GSH) was measured at 412nm, using the reagent DTNB, lipid peroxidation in
plasma was determined at 535 nm in triplicate by the thiobarbituric acid method (TBARS) and protein carbonyl (PC) were determined
using DPNH. Comparisons among the different chagasic groups (IA to III) at the two different conditions (before and after BZN
treatment) were carried out by ANOVA, with post hoc analysis using the Tukey test. Student t-test was used to compare each chagasic
group before and after BZN treatment. Results: After BZN treatment, catalase showed increased activity in groups IA (Bef. BZN: 8.84
0.87; After BZN: 11.96 0.74), IB (Bef. BZN: 6.95 0.41; After BZN: 10.76 0.58) and II (Bef. BZN: 9.34 1.26; After BZN: 13.47
1.31) respectively, while the increase in group III was not significant (Bef. BZN: 6.91 1.11; After BZN: 9.24 0.91). The activity of
glutathione peroxidase showed increased values in all groups: IA (Bef. BZN: 1.84 0.21; After BZN: 3.39 0.20), IB (Bef. BZN: 1.80
0.14; After BZN: 3.64 0.18), II (Bef. BZN: 2.05 0.33; After BZN: 4.37 0.14) and III (Bef. BZN: 1.59 0.12; After BZN: 3.83
0.28). While the SOD activity showed increases in group IA (Bef. BZN: 108.26 8.42; After BZN: 214.51 5.62), IB (Bef. BZN: 116.15
9.43; After BZN: 181.5 8.85), II (Bef. BZN: 122.24 10.30; After BZN: 195.07 10.66), in group III (Bef. BZN: 129.33 24.47; After
BZN: 188.51 20.84) this difference was not significant. GST showed enhanced significant activities in groups IA (Bef. BZN: 23.59
2.21; After BZN: 52.70 4.09), IB (Bef. BZN: 22.34 1.97; After BZN: 45.88 2.88), II (Bef. BZN: 17.56 1.69; After BZN: 41.65
4.98) and not significant in group III (Bef. BZN: 13.66 3.19; After BZN: 39.36 6.62) after treatment. In the other hand, the activity of
glutathione reductase showed enhanced activity only in group IB (Bef. BZN: 4.33 0.33; After BZN: 5.49 0.22), while in the other
groups no differences were detected. GSH and TBARS levels were not changed and PC levels showed enhanced significant levels in:
group IA (Bef. BZN: 0.11 0.01; After BZN: 0.18 0.01), group IB (Bef. BZN: 0.13 0.01; After BZN: 0.21 0.02), group II (Bef. BZN:
0.12 0.01; After BZN: 0.24 0.03), and not significant differences in group III, in spite of a tendency to increase (Bef. BZN: 0.14
0.01; After BZN: 0.18 0.03). Conclusions: BZN treatment beside the benefit to its tripanocide effect, promoted an oxidative insult in
chronic chagasic patients at different stages of cardiopathy severity. These responses were different from those obtained in rats,
indicating that caution should characterize inferences from murine models regarding BZN effects.

Key: OSD6
Key words: benznidazole, oxidative stress, chagas`s disease

Havana Redox 2009

Oxidative Stress Induction and Antitumoral Effect of Sodium Orthovanadate

TMF Gnther
a
, KB Felipe
a
, EB Parisotto
a
, F Biscaro
a
, A Pereira
a
,

EA Ferreira
a
, M Kviecinsk
a
, KS Moschetta
a
,

R Geremias
b
, D Wilhelm Filho
c
, RC Pedrosa
a

a
Departamento de Bioqumica, CCB-UFSC, Universidade Federal de Santa Catarina, Cidade Universitria, CEP 88040-900,
Florianpolis, Brazil.
b
Departamento de Farmcia, Universidade do Extremo Sul Catarinense, Cricima, Brazil.
c
Departamento de
Ecologia e Zoologia, CCB-UFSC, Universidade Federal de Santa Catarina, Cidade Universitria, CEP 88040-900, Florianpolis, Brazil.

Several studies report that vanadate possesses antineoplasic effects and it was also showed that vanadate
derivatives lead to oxidative stress in animals. The aim of this work was to evaluate the sodium orthovanadate
(SO) antitumor activity in vivo and its potential relationship with free radicals generation. Antitumor activity of
SO was investigated using the Ehrlich ascites carcinoma (EAC) inoculated intraperitoneally in isogenic Balb/c
male mice (~20 g b.w.). The inoculation moment was used to elect the day 0. The experimental treatment
lasted 9 days and on the tenth day animals were sacrificed and the ascitic fluid was collected for the study.
The following parameters were assessed: tumour and packed cell volume, viable cell count, contents of GSH,
lipid peroxidation (TBARS), and protein carbonylation, as well as the activities of catalase, glutathione-S-
transferase and superoxide dismutase. Animals were divided into two groups (n=6): Negative Control (NC),
which received only DMSO 10% (50 L) daily, and the treated group (TG) which received 37.5 mg/kg body
weight per day. Results were expressed by means and standard deviations and were analyzed using one-way
ANOVA and Tukey-Kramer test. A p-value<0.05 was considered statistically significant. SO treatment caused
decreased tumor (4.01.0 mL) and packed cell volume (1.70.4 mL) while it increased the viable cell count
(795 x 10
6
cell/mL) when compared to NC (8.72.4 mL; 3.00.5 mL; 9315 x 10
6
cell/mL, respectively)
demonstrating a SO antitumor potential. The activity of catalase (TG=1.870.06; NC=0.460.03 nmol/mg
protein), SOD (TG=0.120.01; NC=0.100.01 USOD/mg protein), GST (TG=0.120.01; NC=0.100.01
mol/mg protein) and the GSH content (TG=0.710.04; NC=0.380.04 mol/mg protein) were all increased
by the treatment with SO when compared to NC. SO treatment did not cause significant difference at the levels
of lipid peroxidation

and protein carbonyls when compared to controls. The results suggest that SO exhibited
antitumor activity and it also caused induction of oxidative stress in the tumor. These findings can be
considered to indicate a possible SO antitumor action mechanism.

Key: OSD7
Key words: vanadate, oxidative stress, cancer

Havana Redox 2009

Evaluation of the Oxidative Profile in Ehrlich Ascites Carcinoma in Mice
Treated with Cordia Verbenaceae extracts

EB Parisotto
a,b
, KB Felipe
a
, TMF Gnther
a
, EA Ferreira
a
, F Biscaro
a
, MR Kviecinski
a
, A Pereira
a
, AC
Vieira
a
, SRS Ferreira
c
,Correia JFG
a
, R Geremias
d
, D Wilhelm Filho
a
, RC Pedrosa
a

a
b
Depto. de Ecologia e Zoologia,
c
Depto. de Engenharia Qumica e Engenharia de Alimentos. Universidade
Federal de Santa Catarina, Cidade Universitria, CEP 88040-900, Florianpolis, Brazil.
d
Universidade do Extremo Sul Catarinense,
Av. Universitria, CEP 88806-000, Cricima, Brazil

Cordia verbenacea (Boraginaceae) is popularly known in Brazil as erva baleeira, and has been used by
people to treat several diseases, including cancer. Many effective anticancer drugs work generating free
radicals which attack the malignant cells leading to apoptosis. This study aims to evaluate the oxidative effects
of a treatment carried out with the hydroethanol crude extract (HCE) from C. verbenaceae on the Ehrlich
ascites carcinoma (EAC). EAC was inoculated in isogenic Balb/c male mice (~23g b.w.) and this moment was
taken as day 0. Intraperitoneal treatments with HCE (150 mg/kg per day) started on the sixth day and on tenth
day the animals were sacrificed. The ascitic fluid was collected and used for the analysis. Results were
expressed by means and standard deviation and they were analyzed using one-way ANOVA and Tukey-
Kramer test. A p-value<0.05 was considered to be statistically significant. The treatment caused increased
catalase (0.890.08 mol min/ mg protein) and glutamyl-S-transferase activities (0.330.80 mol min/ mg
protein) when compared to the negative control (NC=0.460.03 and 0.190.02 mol min/mg protein,
respectively). No significant differences were observed for the superoxide dismutase activity (HCE=0.070.01;
NC=0.090.05 USOD/ mg protein) and for the levels of lipid peroxidation (HCE=0.090.01; NC=0.070.01
nmol/mg protein). However, levels of GSH and protein oxidation in the tumor were increased importantly
(HCE=0.820.18; NC=0.380.04 mmol/mg protein and HCE=338.24 11.52; NC=33.758.81 mol/mg
protein,respectively). The results showed that HCE exhibited an antitumor potential because it was able to
induce oxidative stress in the tumor fluid.

Key: OSD8
Key words: Cordia verbenacea, oxidative stress, anticancer

Havana Redox 2009

Evaluation of the Antitumor Mechanism of Casearin-Rich Fraction From
Casearia sylvestris Against the Ehrlich Ascites Carcinoma in Mice

a
KB Felipe,
a
TMF Gunther,
a
F Biscaro,
a
EB Parisotto,
a
A Pereira,
a
E Ferreira,
a
M Kviecinski,
b
Correia
JFG
a
, R Geremias,
c
FM Gatti,
c
MH Rossi,
d
D Wilhelm Filho,
a
R C Pedrosa.

a
Departamento de Bioqumica. Universidade Federal de Santa Catarina, Cidade Universitria, CEP 88040-900, Florianpolis, Brazil ;
b
Universidade do Extremo Sul Catarinense, Av. Universitria, CEP 88806-000, Cricima, Brazi;
c
Centro de Sanidade Animal do Instituto
Biolgico de So Paulo;
d
Departamento de Ecologia e Zoologia. Universidade Federal de Santa Catarina, Cidade Universitria, CEP
88040-900, Florianpolis, Brazil.

Casearia sylvestris S.W. (Flacourtiaceae) is a Brazilian medicinal plant known mainly as Guaatonga. It is
rich in casearin, a clerodane diterpene with antitumor bioactivity. Doxorubicin (DOX) is a chemotherapeutic
agent whose action is also due to its ability to generate free radicals. In this study DOX was used as the
positive control (PC) and the action mechanism of the antitumor activity of a fraction rich in casearin (FRC)
was evaluated through comparison. EhrIich ascites carcinoma (EAC) was inoculated in isogenic Balb/c male
mice (~20g b.w.) and this moment was taken as day 0. Intraperitoneal treatments with DOX and FRC
(1,2mg/kg and 150mg/kg per day, respectively) started on the sixth day and on tenth the animals were
sacrificed. Their ascitic fluid was collected and used for the analysis. Results were expresssed by means and
standard deviation and were analyzed using one-way ANOVA and Tukey-Kramer s test. A p-value <0.05 was
considered to be statistically significant. FRC decreased GSH ascitic content
1
when compared to PC (FRC=
0.38 0.12; DOX= 0.89 0.09 mol/mg protein). FRC treatment also decreased the levels of lipid
peroxidation
2
(FRC= 0.05 0.006; DOX= 0.08 0.008 nmol/mg protein), protein oxidation
3
(FRC= 40.95 9.46;
DOX= 77.84 10 mol/mg protein) and GST activity
4
(FRC= 0.17 0.02; DOX= 0.57 0.06 mol min/ mg
protein). No significant difference was observed to catalase activity
5
when the treatments were done with FRC
or DOX (FRC= 0.33 0.05; DOX= 0.38 0.10 mol min/ mg protein). These findings suggest that FRC and
DOX apparently possess different ways of action and the FRC action mechanism does not seen to be the
induction of free radical generation.

References: 1 BEUTLER, E.; DURAN, O et al., 1963 Improved method for the determination of blood glutathione. J Lab Clin Med, v.
61, p. 882-890. 2 OHKAWA, H, 1979 Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem, v. 95, p.
351-358. 3

LEVINE, R. L. et al., 1990. Determination of carbonyl content in oxidatively modified proteins. Meth Enzymol., v. 186, p.
464-478. 4 HABIG, W. H.; PABST, M. J.et al., 1974. Glutathione S-Transferases. The first enzymatic step in mercapturic acid
formation. JBC, v. 249, n. 22, p. 7130-7139. 5 AEBI, H, 1984. Catalase in vitro. Meth. Enzymol., v. 105, p. 121-126.

Key: OSD9
Key words: Casearia sylvestris, oxidative stress, Ehrlich ascites carcinoma, antitumor

Havana Redox 2009

Metallothioneins and Zinc Metabolism in the Prevention and Progression of
the Diabetes Mellitus and their Complications: Experimental Evidence,
Possible Mechanisms and Clinical Implications.
Torres A.
Instituto de Farmacia y Alimentos, Universidad de la Habana. San Lzaro y L, Ciudad Habana, Cuba. Email: amarilis@ifal.uh.cu,
amarilyst@gmail.com

Diabetes is a serious problem of health for its high prevalence and its complications. It has been observed at
experimental level the incidence of the reactive oxygen species in the pancreatic cells damage and in micro-
macro vascular complications of the Diabetes Mellitus. Several lines of evidence point to Zn and its
metabolism as important players in the onset and/or progression of multifactorial diseases. Particularly there
are numerous investigations related with the Zn and Metallothioneins role in the physiopathology of Diabetes
and its vascular complications. Zinc is involved in virtually all aspects of cellular and molecular biology of the
cell as a catalytic, structural, and regulatory cofactor in over 300 proteins. Zinc ions have an insulin-like
(insulinomimetic) effect. Zinc deficiency activates stress pathways and may result in a loss of tyrosine
phosphatase control, thereby causing insulin resistance. The Metallothioneins (MT) are intracellular metal-
binding and cysteine-rich proteins that act mainly as regulator of metal homeostasis such as those of zinc and
copper in tissues. MT were found to be potent antioxidant proteins that protect cells and tissues from oxidative
stress. Genetically or pharmacologically enhanced MT expression in various organs including heart and kidney
provided significant protection from diabetes-induced organ dysfunction such as cardiomyopathy and
nephropathy. This study describe MTs biochemical features and role of MT together with zinc in the
prevention of diabetes and its complications with emphasis on experimental evidence, possible mechanisms,
clinical implications and future perspectives of pharmacological investigations on this disease.

Key: OSD10
Key words: diabetes, metallothionein, zinc

Havana Redox 2009

Evaluation of the A2 Phospholipase Activity in Hypertensive and
Normotensive Patients

Danay Heredia Ruiz, Mildrey Vales Almodovar, Douglas Fernndez Caraballo, Marianela Ballesteros
Hernndez, Jess Alfonso Rodrguez.

Superior institute of medical sciences " Dr. Serafn Ruiz of Zrate Ruiz ". Villa Clara. Cuba. danayhr@iscm.vcl.sld.cu

Introduction: In the pathogenesis of the arterial hypertension (HTA) the increment of the Reactive Oxygen
Species (ROS) plays an important role. The peroxidation of arachidonic acid forms the isoprostanes, a well
known vasoconstricting agents that have been implicated in endothelial dysfunction.
Objective: To compare the activity of the FLA2 enzyme in individual normotensive and hypertensive, belonging
to areas of health from Santa Clara municipality. Materials and methods: There were used blood samples from
patients belonging to the health area of Santa Clara municipality and from HTA consultation of the ISCM-VC. It
was employed a colorimetric technique based on the catalytic action of phospholipase enzyme A2, that produce
an acidification of the halfback due to the liberation of lysophospholipids and fatty acids. These changes in the
pH of the halfback produce changes in the absorbance of the substrate solutions. Results: For the evaluation of
the differences between both groups in relation to the enzymatic activities of FLA2 the student t test was used.
Significant differences were found between the groups. The A2 phospholipase actuvuty was higher in the
hypertensive patients.

Key: OSD11
Key words: phospholipase A2, hypertension

Havana Redox 2009

Evaluation of the Antioxidant Enzymes Activity in Hypertensive and
Normotensive Patients

Mildrey Vales Almodovar, Danay Heredia Ruiz, Douglas Fernndez Caraballo, Marianela Ballesteros
Hernndez, Jess Alfonso Rodrguez.

*Superior institute of medical sciences " Dr. Serafn Ruiz of Zrate Ruiz ". Villa Clara. Cuba. E.Mail: mildreyva@iscm.vcl.sld.cu

Introduction: The increment in the reactive oxygen species (RSO) in arterial hypertension (HTA) has been
attributed to cause endothelial dysfunction doe to the inactivation of the nitric oxide and as consequence, the
diminution in the levels of endogenous antioxidants like vitamin E, reduced glutathion (GSH) and Superoxide
Dismutase (SOD). Objective: To compare the activity of SOD and CAT enzymes, and the seric concentration of
GHS between normotensive and hypertensive patients, belonging to the areas of health from Santa Clara
municipality. Materials and methods: UV-VIS spectrofhotometer was used for the antioxidants determination in
10 cc of venous blood collected from patients. Results: It was verified a marked difference in the levels of the
antioxidant enzymes of hypertensive patients when compared with the normotensive values. It was observed a
significant increase in the SOD and CAT activity and a marked diminution in the GSH concentration in blood
samples from hypertensive patients. It is conclude that there is a noticeable redox unbalance in hypertensive
patients.

Key: OSD12
Key words: hypertension, antioxidant enzymes, glutathione


Havana Redox 2009

Oxidative Damage Markers and DNA Repair Capacity in Two Von Hippel
Lindau Familial Cases.

Gutierrez R, Rivern G, Pupo JB, Pereira N, Pandolfi A, Lantigua A

National Center of Medical Genetics. Higher Institute of Medical Sciences of Havana.
Address: 146 No. 3102. Playa, PC 10600, Havana City, Cuba. Phone: (53) (7) (208) 9991 - 9999 ext.1093.

Introduction: Von Hippel-Lindau disease (VHL) is a rare, genetic multi-system disorder characterized by the
abnormal growth of tumors in certain parts of the body. Individuals with VHL are also at a higher risk than
normals for certain types of cancer, especially kidney cancer. Symptoms of VHL vary among patients and
depend on the size and location of the tumors. Death is usually caused by complications of brain tumors or
kidney cancer. In this study we determine some oxidative stress biomarkers and the repair capacity H
2
O
2
-
induced DNA damage associated with VHL. Materials and Methods. Two patients with VHL and control
subjects were studied. We determined plasmatic levels of Malondialdehyde (MDA) and advanced oxidation
protein products (AOPP) by spectrophotometric methods. We used the alkaline Comet assay to evaluate
baseline oxidative damage to DNA and the repair capacity H2O2-induced DNA damage by kinetics studies.
Results and Conclusions. Our results showed that VHL patients have higher oxidative damage to lipids,
proteins and DNA than controls. The DNA repair capacity had seriously decreased in VHL patients while the
same damage was repaired in a short time in control subjects. Results from these studies suggest that
elevated oxidative damage and less efficient DNA repair capacity may be associated with VHL disease. Larger
casecontrol and follow-up studies are warranted to further test the potential application of the oxidative stress
markers and alkaline Comet assay in VHL risk assessment and prevention.

Key: OSD13
Key words: Hippel-Lindau disease, oxidative stress, DNA


Havana Redox 2009


Sensibility of LDL to Oxidation in the Nefrotic Syndrome.

E Moreno
a
, E Pedroso
b
, D Alonso
b
, Y Alarcn
b
, Y Rubio

a
Medical School Dr. Carlos Juan Finlay.
b
Local Pediatric Teaching Hospital Dr. Eduardo Agramonte Pia- Camaguey, Cuba.

A lowering in the levels of the osmotic pressure conditioned by the hypoalbuminemia present in patients who
suffer from Nefrotic Syndrome, seems to stimulate the hepatic synthesis of lipoproteins which have abnormal
structure with higher sensitivity to the oxidation than native lipoprotein. LDLox induces apoptosis in
macrophages and endothelial cells, which provides an oxidant stress situation that influences the response of
the kid to the therapy. To demonstrate the sensitivity of the LDL to oxidation in kids suffering from uncontrolled
Nefrotic Syndrome and determine the relation between the levels of cholesterol and LDLox, lipid profiles were
compared under informed consent in three groups of patients aging from 5 to 13 years. Group 1, n=20,
supposedly healthy kids. Group 2: n=20, supposedly healthy kids with high cholesterol levels. Group 3: n=15,
kids suffering from uncontrolled Nefrotic Syndrome. Patients who presented associated diseases or were
hospitalized or under vitamins treatment were excluded from the sample. Seric levels of total cholesterol were
determined, triglicerides, HDL Cholesterol, LDL ox and the results were compared with those of the control
group by means of proportion hypothesis test and the distribution of absolute and relative frequency. In group
2 HDL cholesterol and LDLox levels were normal in 100% of cases, even though kids presented high total
cholesterol levels. Group 3 presented high levels of total cholesterol, triglicerids and LDL ox in all cases, with
normal levels of HDL cholesterol, which shows the huge sensitivity of LDL to oxidation in patients suffering
from Nefrotic Syndrome.

Key: OSD14
Key words: LDL, oxidative stress, nefrotic syndrome


Havana Redox 2009

Evaluation of Some Oxidative Stress Biomarkers in Pediatric Down
Syndrome Patients.

Rivern G, Pupo JB, Martinez O, Martinez A, Lantigua A, Pereira N, Gutierrez R, Pandolfi. A.

National Center of Medical Genetics. Higher Institute of Medical Sciences of Havana. Address: 146 No. 3102. Playa, PC 10600,
Havana City, Cuba. Phone: (53) (7) (208) 9991 - 9999 ext.1093.

Introduction: Down syndrome (DS) is caused by trisomy of chromosome 21 and it is characterized by mental
retardation, immunodeficiency, cataracts, increased incidence of leukemia, signs of premature aging, and
neuropathological alterations similar to those found in Alzheimer disease. The mechanisms by which trisomy
21 leads to its characteristic phenotype are unclear. It has been suggested that the pathogenesis of Down's
syndrome involves reactive oxygen species. Objective: In DS patients to determine some oxidative stress
biomarkers. Materials and Methods. The universe of study included 47 children (0-5 years), 11 SD patients
and 36 control subjects, from Havana City. It were determined plasmatic levels of Malondialdehyde (MDA),
Advanced Oxidation Protein Products (AOPP), Total Thiols (SH) and intraerythrocytic enzymatic activities of
Cu/Zn Superoxide Dismutase, Catalase and Glutathione Reductase by spectrophotometric methods. Results
and Conclusions. Our results showed an increased tendency to oxidative damage in DS patients in
comparison with the control group. Besides it were found statistically significant increases in Cu/Zn Superoxide
Dismutase activity and high plasmatic levels of total thiols in DS group. The activity of the others enzymes
tested and the ratio of Cu/Zn Superoxide Dismutase to Catalase in the DS patients were not different from the
control group. It was obtained an increase in Cu/Zn Superoxide Dismutase activity in pediatric Down
Syndromes patients, confirming that reactive oxygen species could be involved in the pathogenesis of this
syndrome.

Key: OSD15
Key words: Trisomy 21, Down Syndrome, reactive oxygen species, oxidative damage, oxidative
stress biomarkers.


Havana Redox 2009

Oligo and Macroelements Determination on Patients Classified as
Normotensive Using Sustained Weight Test

Prez M
a
, Ballesteros M
b
, Prez de Arma A
b
, Mollineda A
c
.

a
Institute of Medical Sciences Dr. Serafn Ruiz de Zrate Ruiz. Vicerrectorate of investigations. Acueduct Rd. and Circunvalation.
Santa Clara, Villa Clara, Cuba. E-mail: maykelperez2003@yahoo.es, maykelpm@iscm.vcl.sld.cu
b
Institute of Medical Sciences Dr. Serafn Ruiz de Zrate Ruiz. Department of Physiology. Santa Clara, Villa Clara, Cuba.
c
Universidad Central Marta Abreu of Las Villas, Camajuan Rd. km 71/2. Santa Clara, Villa Clara, Cuba

Recently hypertension has been considered as a metabolic and structural (genetic and acquired) abnormalities
Syndrome. Within the metabolic one, evidences of oxidative stress participation in its genesis have emerged.
Some oligo and macroelement are very important in oxidative stress status as pro or antioxidants. Our study
proposes the determination of the Normal levels of oligo and macroelements in normotensive patients. Blood
pressure was measured in some working places of Santa Clara (Chemical Bioactive Centre, Milk Products
Enterprise and Territorial CITMA Delegation) obtaining a sample of 130 individuals classified as normotensive
using the sustained weight test, which is a variant of the isometric test, which has showed high usefulness as a
diagnosis methods of this conditions. All patients were measured for sera sodium, potassium, calcium,
magnesium, cupper of zinc, using atomic adsorption spectrophotometers methods. There was no different
regarding sex neither to the normal values found in literature. It is concluded that oligo and macroelement
tested values are inside the normal rank reported for normotensive patients.

Key: OSD16
Key words: oligoelements, macroelements, normotense


Havana Redox 2009


Relationship of the Reactive Oxygen Species with Cervicouterine Cancer

Irene Alonso Camaraza
a
, Mara Teresa Daz Soto
b

a
Policlnico Pedro Espern, Bauta
b
Food Science and Pharmacy College, University of Havana, Cuba.

Cancer is one of the leading causes of death worldwide. In Cuba it is possible the detection of cervicuterine
cancer at early stage by mean of the cytological test at the primary care level. In this way decrease the
mortality and increase the quality of the woman's life. Another important factor related to the pathogenesis of
uterine cancer is the human papilome virus (HPV) that is also a public health problem since it is a sexual
transmission illness. The HPV conditions the loss of the functions of some key process of the cellular
regulation. The oncoproteins of HPV forms complexes and inactivate proteins responsible for the reparation of
the DNA and the apoptosis induction when the mechanisms of repairs fail. This does facilitate the formation of
tumor cells. Among these processes, those related with the mechanisms of redox balance, are involved.

Key: OSD17
Key words: cancer, oxidative stress


Havana Redox 2009

Oxidative Stress and Quality of Life.

Mara Elena Licea Surez, Mara Teresa Daz Soto


The conditions of life connected to the satisfaction of the persons and the personal satisfaction pondered by
the scale of values, aspirations and personal expectations is what is known as quality of life. It has been
proven in the last years, the participation of the ROS in chronic non transmissible affectations (as diabetes)
and the obesity that which implies that, if it is not carried out an appropriate preventive treatment or
appropriate lifestyles are developed, the quality of life of these patients' is affected. In this work it was proven
that in patients altered metabolically there is an oxidative stress and however, they don't use an antioxidant
therapy.
This investigation was carry out to demonstrate the presence of the ROS in patient with some types of chronic
illnesses and to analyze the necessity of more appropriate therapies.
The present study was carried out in a group of patients that suffered metabolic alterations (with high levels of
cholesterol, triglycerides and glucose) and a group control of healthy persons. The levels of important
biochemical parameters were determined.
A significant increase was observed (p = 0.05) in the concentrations of CAT and MDA in sick patients when
compared to healthy controls.

Key: OSD18
Key words: quality of life, oxidative stress


Havana Redox 2009

Determination of Oxidative Stress Markers in a Model of Experimental
Autoimmune Encephalomyelitis for the Study of Multiple Sclerosis

Nielsen Lagumersindez Denis, Maite Elisa Oviedo Glvez, Alain Valdivia Acosta, Gregorio Martnez Snchez.

Food Science and Pharmacy College, University of Havana, Cuba. E.mail: nielsen@cieb.sld.cu

Multiple sclerosis (MS) is one of the most common neurological diseases of young adults in Europe and North
America. Worldwide, approximately 1 million people are afflicted by this chronic inflammatory disease of the
central nervous system (CNS). There are so many variants of experimental autoimmune encephalomyelitis
(EAE), the prime animal model of MS. Each one reflects given subtypes better than others and thus offer
themselves for the investigation of specific questions with respect to immune pathogenesis, lesion evolution,
and therapeutic developments. Increasing evidence shows that oxidative stress plays an important role in the
pathogenesis of MS and its animal model, EAE. The aim of the present work was to study the status of an
extensive array of oxidative stress indicators for a better characterization of the EAE model. The behavior of
the different biochemical markers that indicate bio-molecule damage, the total antioxidant activity in the serum
and rat brain homogenate antioxidant enzyme activity were evaluated. A significant up-regulation of the
markers of bio-molecule damage (malonildialdehyde: MDA for lipids and, advanced oxidation protein products:
AOPP for proteins) were observed when the EAE is induced, suggesting oxidative damage due to the
immunological events that produce the clinical symptoms. Also, a significative (p< 0.05) modification on the
variables indicating total antioxidant state such as the global indicator susceptibility to lipid peroxidation (PP),
and ferric reducing ability of plasma (FRAP), were noted. These indicators showed its value as biomarkers of
the inflammatory and oxidative process that takes place after the EAE induction.

Key: OSD19
Key words: Encephalomyelitis, oxidative stress, multiple sclerosis


Havana Redox 2009

Antioxidant Therapy in Kids Suffering from Congenital Cardiopaties Related
to Cyanosis.

D Alonso
a
, E Pedroso
a
, E Moreno
b
, Y Alarcn
a

a
Local Pediatric Teaching Hospital Dr. Eduardo Agramonte Pia-
b
Medical School Dr. Carlos Juan Finlay. Camaguey, Cuba.

Congenital cardiopaties constitute processes in whose physiopathology we can find free radicals causing
severe damage to the miocardial structure. These free radicals involve a sequence of events associated to the
loss of the membrane integrity. Early intervention in the process of oxidant stress can result in therapeutic
achievements with large benefits in the rates of cardiovascular morbidity. That is why we decided to
demonstrate the importance of antioxidant therapy in kids suffering from congenital cardiopaties related to
cyanosis. Twenty ambulatory kids were chosen between the ages of 4 to 12 years, who suffered from
congenital cardiopaties related to cyanosis. Patients who presented associated malformations or were under
vitamins treatment were excluded from the sample. Under the informed consent of the selected group, the
seric levels of antioxidants was tested: albumin, ceruloplasmin, total bilirubin, uric acid, ascorbic acid, LDLox
and malonildialdehyde. Vitamin E (400mg every 24 hours) was supplied for three months and the same tests
were repeated every month.Data was processed by means of descriptive statistic methods with absolute and
relative frequency distribution. LDLox and malonildialdehyde presented in all the cases over its normal levels
before the treatment was started and in the last month they went back to normal. 97% of the cases suffered
from hyperbilirubinemia and 46% from hyperuricemia. Both parameters diminished considerably after
treatment. Levels of ceruloplasmin were low in 78% of cases and they increased up to the third month, which
demonstrated the presence of antioxidant stress in kids suffering from congenital cardiopaties related to
cyanosis and the need to include in the treatment an antioxidant medication such as Vitamin E.

Key: OSD20
Key words: Antioxidant, cardiopaties, cyanosis, oxidative stress

Havana Redox 2009

Immunological and Oxidative Stress Mechanisms Involved in the
Recombinant Alpha IFN Treatment in Multiple Sclerosis Patients.

Pentn-Rol G
a,
Cervantes-Llanos M
a
, Martnez-Snchez G
b
, Cabrera-Gmez JA
c
, Valenzuela-Silva
CM
a
, Ramrez-Nez O
b
, Casanova-Orta M
b
, Gmez-Chvez D
a
, Lpez Saura P
a
.

a
Clinical Trials Division. Center for Genetic Engineering and Biotechnology,
b
Center for Research and Biological Evaluations, Institute
of Pharmacy and Food Sciences, University of Havana,
c
International Center of Neurological Restoration.

Type I Interferons (IFN-alpha/beta) were the first immunomodulatory agents to show clinical efficacy for
Multiple Sclerosis (MS) treatment, as a disease modifying therapy. Human IFNs- alpha/beta interact with the
alpha IFN receptor. The interaction with the receptor components results in the activation of a signalling
pathway leading to the regulation of specific genes and proteins which contributes to their numerous biological
effects. There is increasing evidence that MS is not only characterized by immune mediated inflammatory
reactions but also by neurodegenerative processes. In neurodegenerative diseases, neuronal and axonal loss
is mediated by oxidative stress and excitotoxicity which constitute a final common toxic pathway. In our study,
we evaluated the possible molecular mechanisms involved in the in vivo effects of the recombinant alpha-2b
IFN (r-alpha-2b IFN) treatment during 6 months in relapsing-remitting MS patients (RR-MS). We compared the
modulation of genes involved at different steps of MS pathogenesis: antigen presentation, Th1/Th2 cytokines
expression, brain blood barrier (BBB) integrity, regulatory T cells and the serum levels of oxidative stress
markers. Total RNA from peripheral blood mononuclear cells of rIFN-a2b treated MS patients and untreated
MS patients, was extracted and amplified for CD86, CD28, CTLA-4, TNF-, IFN-, CCL2, CCR5, IL-13, MMP-
9, TIMP-1, CD25, TGF-, IL-10, and the transcriptional factor Foxp3 by Reverse Transcription- Polymerase
Chain Reaction. Also, serum levels of oxidative stress markers: malondialdehyde (MDA), advanced oxidation
protein products (AOPP), peroxidation potential (PP), superoxide dismutase (SOD), catalase (CAT), and total
hydroperoxides (THP) from MS treated patients and untreated patients were measured. We found a significant
down-regulation of the CD28 costimulatory signal (p=0.043) and MMP-9 metalloproteinase (p=0.043) in the
group of patients treated with r-alpha-2b IFN. Furthermore, the chemokine CCL2 (p=0.011) and the regulatory
cytokine TGF-beta were up-regulated with the treatment as well (p=0.036). There were no differences
concerning the modulation of the genes studied, in untreated patients. Additionally, ours results showed that
MS is characterized by a marked REDOX imbalance favoring a pro-oxidant environment (statistically
significant for almost all REDOX markers) however r-alpha-2b therapy did not modified oxidative stress
markers. These finding strongly suggest that well-designed clinical studies are needed in order to assess
whether the antioxidant therapy together with other conventional treatments might be beneficial for MS
disease.

Key: OSD21
Key words: oxidative stress, interferons, multiple sclerosis, immunomodulation


Havana Redox 2009

Determination of Oxidative Stress Markers in Patients with Cardiovascular
Diseases.

Livan Delgado Roche
a
, Gregorio Martnez Snchez
a
, Arqumidez L. Daz Batista
b
.

a
Center of Studies for Research and Biological Evaluations (CEIEB-IFAL-UH), Institute of Pharmacy and Food Science, Havana
University, San Lzaro y L, Havana 10 400, Cuba. E-Mail: ceieb@cieb.sld.cu
b
National Institute of Angiology and Cardiovascular Surgery. Calzada del Cerro No.1551, La Habana, E-Mail: arquidia@infomed.sld.cu

The aim of this study was to investigate the status of an extensive array of redox indices in patients
with Cardiovascular Diseases compared with healthy subjects. Blood samples from 113 patients and
60 healthy subjects were tested by spectrophotometric techniques in order to measure oxidative
stress indices. All measured biomarkers of antioxidants/pro-oxidants balance were significantly (p <
0.05) modified in patients compared with normal subjects. An increase of biomolecule damage
markers was noted, a significantly (p< 0.05) reduction of the soluble antioxidant glutathione,
antioxidant enzymes were activated in these patients with a disruption of the catalase/superoxide
dismutase balance compared to normal subjects. In addition, the global indicator susceptibility to lipid
peroxidation was increased in 73.02 % in patients compared to the control group. Further, the
indicator of total antioxidant capacity was significant (p< 0.05) reduced in these patients. The
evaluated indicators could be extrapolated to routine clinical analysis and contribute to an integral
overview of the redox enviroment in patients with Cardiovascular Diseases and could also be used as
indices of treatment efficacy.

Key: OSD22
Key words: oxidative stress, interferon, cardiovascular diseases, clinic


Havana Redox 2009

Redox Salival Environment. Comparison Between Patients Affected by
Inflammatory Periodontal Disease and Periodontal Healthy Patients.

Hetzel de la C. Lourido Prez
a
b
, Gema Prez Davison
b

a
Clnica Estomatolgica Provincial Docente Antonio Briones Montoto, Calle Sol s/n. Pinar del Ro, Cuba. E.mail:
hetzel@princesa.pri.sld.cu
b
CEIEB-Instituto de Farmacia y Alimentos, La Habana, Cuba.

The aim of this study was to assay the saliva redox environment in patients affected by inflammatory
periodontal disease and periodontal healthy controls. It was made an observational, transversal study of cases
and controls in Pinar del Ro, between July 2006 and July 2007. The total sample was 35 individuals; two
groups were conformed, Study with affected patients, and Control with periodontal healthy controls; both of
them were interrogated and oral examination was made; all the information was registered in a form. Saliva
samples were collected and refrigerated at -20
o
C after the biochemical determinations were made in HITACHI
spectrophotometer. The results were analyzed using Outliers test, Shapiro-Wilks test, T-Student test and
Pearsons correlation coefficient. The statistical signification level was p < 0.05. It was verified an unbalance in
redox environment of periodontal affected persons respect healthy controls. It was found significant higher
values of Advanced Oxidation Protein Products in affected patients respect healthy controls and values
significant higher in Ferric Reducing Ability of Plasma in healthy controls with respect to affected patients. It
was determined that Advanced Oxidation Protein Products and Ferric Reducing Ability of Plasma were the
variables with more diagnostic values in this pathology.

Key: OSD23
Key words: inflammatory peridontal diseases, clinic, oxidative stress


Havana Redox 2009

Presence of Oxidative Stress in Preclinical Model of Alcoholism.

Mara Teresa Daz Soto, ngela Fraga, Mara de los ngeles Bcquer


The alcoholism is a toxicomanic that affects the individual that suffers it so much like to those that surround it,
with economic and social implications. The therapy used in the alcoholic patient causes adverse reactions that
commit the quality of life of the patient one and it doesn't control the addictive behavior. The present study has
as objective to obtain a model preclinical of alcoholism and to establish relationship between the addictive
behavior and the presence of oxidative stress to carry out later studies with anti-oxidant therapy. The study
was carried out with 20 rats male Lewis of 200 g of weight divided in two groups: 10 controls and 10 tried with
ethanol. The investigation was divided in 4 stages: Learning, administration of alcohol (5%, 10%, 20%, 30%,
40% at libitum, each concentration during 2 weeks), pharmacological tolerance, abstinence. They were carried
out behavioral tests and biochemical determinations in each stage. The results demonstrated that in the
measure that the rats acquired the addiction for the alcohol it diminished the capacity by heart and function
motorboat. As for the parameters related with the state redox in the alcoholic rats increase of MDA existed
(Malonildiladehide) and (PP) peroxidacin Potential with regard to rats controls. The analysis of the results
allows to conclude that the gradual administration of ethanol to different concentrations to rats Lewis allows to
develop a model of alcoholism in which you/they are related the behavioral dysfunctions and the oxidative
stres.

Key: OSD24
Key words: alcoholism, oxidative stress


Havana Redox 2009

Determination of the Reactive Oxygen Species in type 2 Diabetic Patients

Caridad Margarita Garca Pea
a
, Mara Teresa Daz Soto
b

a
Centro de Investigacin y Desarrollo de Medicamentos, Ave. 26 # 1605 e/ Boyeros y Puentes Grandes, CP. 10600, Cuba.
b
Centro de
Estudios para las investigaciones y Evaluaciones Biolgicas, Instituto de Farmacia y Alimentos, Universidad de la Habana, Cuba

It has been proved that Reactive Oxygen Species (ROS) participate in the destruction of the -pancreatic
cells, which influences in Diabetes Mellitus (DM). A study of the changeable indicators of oxidizing stress
determining the Superxide Dismutase Enzymes (SOD), catalase (CAT), levels of malonil dialdhyde (MDA) and
Fructosamine. It was found a diminishing in the activity of SOD and an increase in CAT and lipidic peroxidation
expressed as MDA in patients with Tipe 2 Mellitus Diabetes. It was found a correlation between fructosamin
and MDA and between SOD and MDA in patients with Diabetes Mellitus Type 2, suggesting that there is a
relation with products of advanced glicosilation and the generation of superoxid anion and hydrogen peroxide.
These results show the presence of oxidative stress in patients with Diabetes Mellitus Type 2.

Key: OSD25
Key words: diabetes, oxidative stress


Havana Redox 2009

Oxidant Stress in Congenital Heart Diseases. Clinical - Lab Diagnosis.

E Pedroso
a
, D Alonso
a
, E Moreno
b
, Y Alarcn
b
, CM Oliva
a

a
Paediatric Teaching Hospital Dr. Eduardo Agramonte Pia-
b
Medical School Dr. Carlos J. Finlay. Camagey, Cuba

Fifty one percent of all congenital anomalies have their origin in cardiopathies and the rate of incidence is 6 to
8 every one thousand live births. Ischemic heart disease constitute the manifestation of a process that begins
with an excess of free radicals which initiates the atherosclerotic process due to the damaged caused to the
vascular walls, provoking penetration into the subendotelial space of the low density lipoproteins (LDL) and as
a result to the atherosclerotic plaque. Lead by the purpose of diagnosing oxidant stress in children, we took
two groups of patients diagnosed with cyanosis congenital heart disease related to: (group 1: n=20); acyanotic
congenital heart disease (group 2: n=30), and a third control group of supposedly healthy kids (group 3: n=50).
All of them were four to twelve years old. Patients who presented associated malformations or were under
vitamins treatment were excluded from the sample. To check an the antioxidant capacity an the levels of
LDLox as a pointer of oxidant stress, comparing the results with the control group through the distribution of
relative and absolute frequency and proportion hypothesis test with a 95 % of significance level. In the first
group high levels of bilirubin were detected (97 %), uric acid (46 %), and diminished ceruloplasmin (78 %). In
the second group hiperuricemia was present in the 94 % of cases and ceruloplasmin diminished in 14 % of
patients. In both groups, LDLox increased in 100 % of the cases, which proves the diagnosis of oxidants stress
in children.

Key: OSD26
Key words: oxidative stress, congenital heart disease


Havana Redox 2009

Oxidative Stress and Its Impact in Cataracts

F Yaheln
a
, M Gregorio
a
, W Damin, T Nantembwa

Pharmacy and Foods Institute. University of Havana. Ave. 23 No. 21425 e/ 214 and 222. La Coronela. La Lisa

Cataracts are a serious health problem mainly in older pacient, its incidence in the population older than 65
years is approximately of 25 percent and in people older than 80 years its over 50 percent. This situation is
particularly serious in the developing countries where 17 million persons are blind due to cataracts. In Cuba
the pathology incidence is also high, however the number of persons becoming completely blind is scarce
because of the Cuban National Health System. For example, in The Ophthalmological Complex Pando Ferrer
of Havana City doctors carry out an average of 4000 surgeries per year. In order to strike out lens opacity,
which is the fundamental cause of blindness in cataracts. Some risk factors have been identified in this
affection nowadays free radicals and oxidative stress have been suggested to be part of this process. In this
work, a review took place with the purpose to establish most important achievements related to the
development of cataracts and its relation with oxidative stress. That is why bibliographies sources dating since
2000 until now were checked. The results show that cataract is related with reactive oxygen species
generation; the research also proved the preventive use of antioxidants that delay the start of the pathology
symptoms.

Key: OSD27
Key words: oxidative stress, cataracts


Havana Redox 2009


Oxidant Stress in Kids Suffering from Diabetes Mellitus Type 1.

Y Rubio
a
, E Pedroso
a
, D Alonso
a
, E Moreno
b
, Y Alarcn
a

a
Local Pediatric Teaching Hospital Dr. Eduardo Agramonte Pia-
b
Medical School Dr. Carlos Juan Finlay. Camaguey, Cuba

Diabetes mellitus is a progressive disease with a high risk of mortality which affects more than 30 millions of
persons worldwide. The main cause of death in these patients is a series of complications that involves blood
vessels, higher lipid levels, an increase on the oxidation of LDL and a growth in the platelet aggregation. The
pathogenesis of this disease is also related to free radicals and alterations due to glucosilations of the
structure of the LDL, which turns it into a particle more leaning to oxidation. That is why it was decided to
diagnose oxidant stress in kids suffering from Diabetes mellitus type 1. Thirty ambulatory patients were
selected between 6 to 12 years of age, who had been suffering from Diabetes mellitus type 1 since 3 years
ago. Patients who presented uncontrolled associated diseases or were hospitalized or under vitamins
treatment were excluded from the sample. Under informed consent were determined the seric levels of
glucose, albumin, ceruloplasmin, total bilirrubin, uric acid, ascorbic acid and LDLox as a pointer of oxidant
damage. Data was processed by means of descryptive statistic methods with absolute and relative frequency
distribution. 93% of patients presented slightly higher levels of glicemic, diminished levels of vitamin C (33%),
ceruloplasmin (17%), albumin (63%). LDLox levels increased in all kids which shows the presence of oxidant
stress in these kids.

Key: OSD28
Key words: oxidative stress, diabetes


Havana Redox 2009

T-cell Immunoregulation and Oxidative Stress in Neuromyelitis Optica.

Cervantes-Llanos M
a
, Martnez-Snchez G
b
, Cabrera-Gmez JA
c
, Valenzuela-Silva CM
a
, Ramrez-
Nuez O
b
, Casanova-Orta M
b
, Lopategui-Cabezas I
d
, Lpez Saura P
a
, Pentn-Rol G
a
.

a
Clinical Trials Division. Center for Genetic Engineering and Biotechnology,
b
Center for Research and Biological Evaluations, Institute
of Pharmacy and Food Sciences, University of Havana,
c
International Center of Neurological Restoration,
d
Higher Institute of Medical
Sciences Victoria de Girn.

Neuromyelitis optica (NMO) is an idiopathic central nervous system (CNS) demyelinating syndrome although
the cause of the disorder is not known; several lines of evidence suggest that the fundamental immunological
process is driven by humoral mechanisms. Clinical experience implies that plasmapheresis and
immunosuppressive therapies are beneficial for treatment and prevention of acute attacks however the
standard immunomodulatory drugs may not alter the course of NMO suggesting the involvement of other
factors in the NMO pathogenesis. Conversely to humoral immune arm, cytokines, chemokines related to the
cellular immune response, the blood barrier integrity markers and oxidative stress component have been
poorly studied. We performed a molecular characterization of cellular immune response and oxidative stress
parameters in serum from NMO patients. We measured the serum levels of TNF-, IFN-, IL-10, MMP-9
metalloproteinase, its inhibitor TIMP-1 and oxidative stress markers: malondialdehyde, advanced oxidation
protein products (AOPP), peroxidation potential, superoxide dismutase, catalase, and total hydroperoxides
from NMO patients and healthy controls. Serum levels of TNF- and IL-10 cytokines from NMO patients were
down regulated suggesting that in addition to a humoral disregulation, an effector - regulator imbalance could
be imply in its pathogenesis. Furthermore, we detected an up regulation statistically significant of all
parameters of oxidative stress studied indicating a noteworthy redox imbalance mainly due to an up-regulation
of the SOD activity and a low CAT activation. This pro-oxidant environment reflected on the elevation of the
lipid peroxidation and the up-regulation of the AOPP, evidence the chronic inflammatory process taking place
in NMO patients for which an antioxidant as much as an immunomodulatory therapy is required.

Key: OSD29
Key words: oxidative stress, neuromyelitis optica, immunoregulation


Havana Redox 2009

ApoH Polymorphisms, ApoH Plasma Levels and Oxidative Markers in Type
2 Diabetic Patients.

Antoni Castro
a
, Iolanda Lazaro
a
, Ela Cspedes
b
, Josefa Girona
a
, Montserrat Guardiola
a
, Anna Cabr
a
, Lluis
Masana
a

a
Unidad de Investigaciones en Lpidos y Arteriosclerosis. Facultad de Medicina de Reus. Universidad Rovira i Virgili. Reus-Tarragona,
Espaa
b
Facultad de Medicina Calixto Garca, 16400, Cuba.

The dyslipidemias, hypercoagulability and the oxidative stress have been implicated in the appearance,
progression and severity of the vascular complications that appear in diabetic patients. ApoH is a glycoprotein
involved in the coagulation pathway, and genetic variants determine modification in its activity with possible
repercussion in the oxidative metabolic state. To know the associations between genetic variations in the gene
of this glycoprotein and its biological function we studied Val247Leu and Trp316Ser polymorphisms, plasma
levels of ApoH and lipid and oxidative markers in 169 type 2 diabetic subjects with and without signs of
atherosclerosis. Those carriers for the Leu allele had lower HDL-cholesterol levels and higher maximal diene
production compared to those Val carriers (HDLc: 1.160.27 mmol/l vs 1.280.31 mmol/L, respectively,
p=0.009; and maximal diene production: 663.592.6 mol diene/mol LDL vs 633.978.0 mol diene/mol LDL,
respectively, p=0.037) and a tendency to have higher maximal rate and lower lipoperoxide levels. HDLc, vitamin
E and lipoperoxides were significantly lower in patients with vascular complications and LL, VL carriers in
relationship with VV genotype. Polymorphism Val247Leu was associated with diene formation in diabetic
patients without atherosclerosis. At the same time, ApoH was positively correlated with vitamin E (r=0.208,
p=0.007) and inversely correlated with LDL oxidation susceptibility (lag phase) (r=-0.162, p=0.046). There were
no relationship between ApoH and other markers of oxidation measured in plasma, such as oxidized LDL or
lipoperoxides. The Trp316Ser and Val247Leu ApoH gene variants did not modulate ApoH plasma values in this
population although there are high plasma levels of ApoH in diabetic patients with atherosclerosis

Key: OSD30
Key words: Apo H, oxidative stress, diabetes, atheroesclerosis


Havana Redox 2009


Havana Redox 2009

Redox Disruption in Male Infertility

Rosa Mara Flores Snchez
a
, Celia A. Alonso Rodrguez,
a
Gregorio Martnez Snchez,
b
Akel Mallok
b

a
Hosp. Hermanos Ameijeiras, La Habana Cuba, E.Mail: celialon@infomed.sld.cu,
b
Center For Research and Biological Evaluations,
Havana University, La Habana, Cuba. E.Mail: gregorioms@infomed.sld.cu

Couple's infertility has been recognized as a health-social problem of the reproductive health worldwide. Couples that not
achieve pregnancy after a period of 12 months of sexual activity without using birth-control methods are defined as
infertile couples. The incidence of infertility is notably in different between countries, and even in different areas of oneself
country. Male factor has been considered a major contributory factor to infertility 35%. Along with the conventional causes
for male infertility such as varicocele, cryptorchidism, infections, obstructive lesions, cystic fibrosis, trauma, and tumors, a
new, yet important cause has been identified: oxidative stress. Oxidative stress (OS) is a result of the imbalance between
reactive oxygen species (ROS) and antioxidants in the body, which can lead to sperm damage, deformity and eventually
male infertility. This involves peroxidative damage to sperm membrane and DNA fragmentation at both nuclear and
mitochondrial levels. OS has been implicated as the major etiological factor leading to sperm DNA damage. OS-induced
DNA damage can lead to abnormalities in the offspring including childhood cancer and achondroplasia. In this poster, we
discuss the need of ROS in normal sperm physiology, the mechanism of production of ROS and its pathophysiology in
relation to male reproductive system. ROS are produced by a variety of semen components, Including immotile or
morphologically abnormal spermatozoa, leukocytes, and morphologically normal but functionally abnormal spermatozoa.
One the effects of the excessive ROS production, is the cellular membrane alteration, due to lipid peroxidation (LPO)
which is a very important pathophysiological process occurring in numerous diseases and stress conditions, and it usually
results in a cascade of profound degradative processes, affecting the organization and function of biological membranes.
The membranes of the human spermatozoon contain a high concentration of polyunsaturated fatty acids; therefore they
are susceptible to the LPO damage. ROS have a variety of effects on the spermatic cell, this is a very controversial
subject and it is one of the reasons for this article, reviewing some of the biochemical aspects related to ROS production
and diagnostic methods in male infertility. Oxidative stress results from the production of oxygen radicals in excess of the
antioxidant capacity of the stressed tissue. Many conditions or events associated with male infertility are inducers of
oxidative stress. X-irradiation, for example, or exposure to environmental toxicants and the physical conditions of
varicocele and cryptorchidism have been demonstrated to increase testicular oxidative stress, which leads to an increase
in germ cell apoptosis and subsequent hypospermatogenesis. Such stress conditions can cause changes in the dynamics
of testicular microvascular blood flow, endocrine signaling, and germ cell apoptosis. Testicular oxidative stress appears to
be a common feature in much of what underlies male infertility, which suggests that there may be benefits to developing
better antioxidant therapies for relevant cases of hypospermatogenesis. Although reports have suggested the benefit of
antioxidant treatment of infertile men, many studies also show no effect. Most studies in the literature are not randomized,
placebo controlled, or double blinded in design, which makes it difficult to differentiate regression toward the mean from
true positive treatment effects. The small patient sample sizes and varying male populations also add to the difficulty in
comparing studies. Pregnancy, the most relevant outcome parameter, is rarely reported. Ideally, patients would be
selected based on OS levels, and improvement in these levels would be correlated to improvement in pregnancy rates.
Until those studies are performed, the use of antioxidants for the treatment of male infertility remains empiric.

Key: OSD31
Key words: infertility, oxidative stress


Havana Redox 2009


Oxidative stress and pregnancy

Cardosa E, Trujillo R, Cardosa P, Fernndez Y

Medical School. Guantnamo. Calle 5 West N.N e/ 6 y 9 North. Guantnamo. Email: farma@infosol.gtm.sld.cu

The cardiovascular adaptation in the pregnancy are essentials to secure an appropriate provide of oxygen and
nutrients to the placenta and the fetus. This adaptation include: increase of blood volume, increase of cardiac
output and diminish of peripherical artery resistance which are linked with a high Nitric Oxide production (NO),
probably estimated by a stress friction increaser and the running level of sexual hormones that happen
during the gestation. One of the consequence of such adaptation is constituted by the high blood pressure
and with it the preclampsiu which is the result of a serie of factors that end a physiopathological alteration
very common : an unbalance in the free- radical production. Keeping in mind the severity of this entity so as
the mother as the fetus , we took into account to purpose to deep in the topic because we determined to fix the
following conclusions: the infections and inflammatory process during the pregnancy, could disturbs the
balance of free radicals as well as the preclammpsiu

s origins can link enchange of Arginine- L way, the goal
of the preventive intervention as treatment must be led to restore the relation of ON with O
2
, avoiding the
ONOO production and it toxic effects. As the L- Arginine it is founded in animals protein of high biology value,
like fish, which could be the ingestion of this during the gestation, an alternative to warrant the nutritional
supply of amino acid of this stage.

Key: OSD32
Key words: pregnancy, preeclampsia


Havana Redox 2009


Characterization of oxidative stress in spinocerebellar ataxia type 2 patients

Omar Ramrez Nez,
a
Luis Velzquez-Prez,
b
Gilberto Snchez-Cruz,
b
Mariela Guevara,
c
Gregorio Martnez
Snchez
a

1
Centre for Research and Biological Evaluations, Institute of Pharmacy and Food Sciences, University of Havana, Cuba.
2
Centro para la Investigacin y Rehabilitacin de las Ataxias Hereditarias Carlos J. Finlay Holgun, Cuba.

The role of the reactive oxygen species in neurodegenerative disorders and its complications has been
established. The aim of this study was to investigate the status of an extensive array of redox indices:
glutathione (GSH), malondialdehyde, advanced protein oxidation products, peroxidation potential (PP), Ferric
Reducing Ability of Plasma, superoxide dismutase (SOD), Catalase (CAT) and total organoperoxides levels in
patients with Ataxia compared to healthy subjects. This analysis contributes to establish the reference values
that would permit examination of the role of oxidative stress which causes damage to biomolecules related
with ataxia patients and open the bases to follows the efficiency of future therapies. Blood samples from
spinocerebellar ataxia type 2 (SCA2) patients and healthy subjects were tested by spectrophotometric
techniques in order to measure oxidative stress indices. All measured biomarkers of antioxidant-prooxidant
balance were significantly modified in SCA2 patients compared with normal subjects. It was noted an increase
of markers of damage to biomolecules. A significantly reduction of the soluble antioxidant GSH levels were
also observed in SCA2. Patients enzyme SOD and CAT were activated in these patients with a disruption of
the balance CAT/SOD compared with normal subjects. In addition, the global indicator susceptibility to lipid
peroxidation PP was increased in SCA2 in relation to the control group. The used clinical diagnosis covers
different components making possible an integrative analysis of the redox balance in SCA2 patients. The
evaluated indicators could be extrapolated to the routine clinical analysis and contribute to an integral overview
of the oxidants / antioxidants balance in SCA2 patients and also could be used as indices of treatments
efficacy.

Key: OSD33
Key words: spinocerebellar ataxia; neurodegenerative diseases, biomarkers, free radicals, reactive
oxygen species

Havana Redox 2009

Antioxidants
Index

Antioxidants............................................................................................................................Error! Marcador no definido.
Mitochondrial-Targeted Antioxidants as a Therapy in a Model of Tolerance to Nitroglycerin...........Error! Marcador no
definido.
Antioxidant Capacity Determination in Citric Juice ...........................................................Error! Marcador no definido.
Total Anthocyanins and Total Phenolics of Capulin (Prunnus capuli) and Their Antioxidant PropertiesError! Marcador
no definido.
Antioxidant Activity of Loquat (Eriobotrya japonica)..........................................................Error! Marcador no definido.
Neuroprotective Effect of Aqueous Extract of Yellow Maca in Newly Weaned Male Rat .............................................. 70
Antioxidants and Atherosclerosis: Effect of Hydroalcoholic Extracts of Plants for Medical Uses on Key Processes of
the Atherogenesis.............................................................................................................Error! Marcador no definido.
Vitilvenz

Natural Alternative Therapy in Vitiligo ..............................................................Error! Marcador no definido.
Evaluation of the Antioxidant Potential of Banana (Musa Cavendish, Musaceae) Peel Hydroethanol Crude Extract in a
Wound Healing Model.......................................................................................................Error! Marcador no definido.
Hepatoprotective Effect of 2,4,6-trihydroxiacetophenone Against Oxidative Stress Induced by Carbon Tetrachloride in
Mice ..................................................................................................................................Error! Marcador no definido.
Anthraquinones Extracts From Different Organs of Morinda royoc L. In Vitro Plants and Their Antioxidant Properties.
Antioxidant and Antigenotoxic Evaluation of DI-31, a Brassinosteroid Analogue.............Error! Marcador no definido.
Antioxidant Protection of Vascular Endothelium in Oxidative Damage Models to Cultured Endothelial Cells........ Error!
Content of Redox-Active Compounds in Foods Consumed in Macrobiotic Diet...............Error! Marcador no definido.
Determination of Antioxidant Capability of Drinks of Jamaica and Tamarind ...................Error! Marcador no definido.
Antioxidant and Wound Healing Effect of Rhizophora mangle (L.) in an Aseptic Wounds Model in Rats. .................... 80
Fruits and Vegetables Ingested by Hypertensive Women: Relationship of The Lipidic Profile and The Antioxidant Total
Capacity in Serum.............................................................................................................Error! Marcador no definido.
In vitro Antioxidant Activity and Chemical Composition of Halimeda monile Seaweed Aqueous Extract ............... Error!
Protective Effect of Bidens pilosa L Extract in Hepatotoxicity Induced by Paracetamol. ..Error! Marcador no definido.
Antioxidant Activity and Atheroprotective Effect of an Aqueous Extract of the Marine Seaweed Halimeda incrassata
): Antioxidant Properties in Inflammatory Models ...Error! Marcador no definido.

Piper auritum HBK Cream: Antinflammatory Activity by Antioxidant Mechanisms. ..........Error! Marcador no definido.
Therapeutic Effect of Mangifera indica L. in Minor Recurrent Aphthous Stomatitis..........Error! Marcador no definido.
Antioxidant Activity of Six Aromatics Plants Used in Cuba as Condiments......................Error! Marcador no definido.
Absorption and Metabolism of Flavonoids and Their Metabolites ....................................Error! Marcador no definido.
Isolation of Antioxidant Compounds from Medicinal Extract of Pedilanthus Tithymaloides........................................... 90
Phenylpropanoid Content in Plant of Theobroma cacao L.: Antioxidant Activity ..............Error! Marcador no definido.
Mangifera indica L Extract (Vimang) and Its Main Polyphenol Mangiferin Prevent Mitochondrial Oxidative Stress in
Atherosclerosis-Prone Hypercholesterolemic Mouse .......................................................Error! Marcador no definido.
Protective Effects of Mangifera indica L extract (Vimang), and Its Major Component Mangiferin, on Iron-Induced
Oxidative Damage to Rat Serum and Liver ......................................................................Error! Marcador no definido.
Antioxidant Capacity Determination in Mandarin, Maracuya and Grape Juices...............Error! Marcador no definido.
Effect of Aqueous Extract of Yellow Maca in the Levels of Glutathione in the Brain After an Intense Exercise in Newly
Weaned Male Rat .............................................................................................................Error! Marcador no definido.
Antioxidant Capacity of the Aqueous Extract of Peperomia choroniana D.CD. (ipintaki) by Means of DPPH and
Reception of Hydroxyl Radical..........................................................................................Error! Marcador no definido.
Ferric Reduction Antioxidant Power evaluation on Murraya paniculata L (Jack) tincture. Error! Marcador no definido.

Havana Redox 2009

Biological activity and chemical composition of Morinda citrifolia liin and Annona muricata extracts.... Error! Marcador
no definido.

Havana Redox 2009

Antioxidants
Mitochondrial-Targeted Antioxidants as a Therapy in a Model of Tolerance
to Nitroglycerin
M. Rocha
a,b
, R. Garca-Bou
c
, C. Bauls
b
, J. R. Herance
d
, J. V. Esplugues
c
, A. Hernndez-Mijares
a
and V. M.
Vctor
a,b

a
Service of Endocrinology. Universitary Hospital Dr. Peset. Avda. Gaspar Aguilar 90, 46017 Valencia. Spain.
b
Dr. Peset Hospital Research Foundation. Avda. Gaspar Aguilar 90, 46017 Valencia. Spain
c
Pharmacology Department and Ciberehd, Faculty of Medicine, University of Valencia C/ Blasco Ibaez 15, 46010 Valencia. Spain.
d
Institute of High Technology The Barcelona Biomedical Research Centre, C/ Dr. Aiguader, 88 08033 Barcelona, Spain

Nitroglycerin (GTN)-tolerance was induced in vivo (rats) and in vitro (rat and human vessels).
Electrochemical detection revealed that the incubation dose of GTN (5X10
-6
mol/L) did not cause NO
to be released or modify O
2
consumption when administered acutely. However, development of
tolerance produced a decrease in both mitochondrial O2 consumption and the km for O2 in animal
and human vessels and in endothelial cells in a non-competitive action. GTN tolerance has been
associated with impairment of GTN biotransformation through inhibition of ALDH-2 and with
uncoupling of mitochondrial respiration. Feeding rats with mitochondrial-targeted antioxidants
(mitoquinone, MQ), and in vitro co-incubation with MQ (10
-6
mol/L) or glutathione ester (GSH ester,
10
-4
mol/L) prevented tolerance and the effects of GTN on mitochondrial respiration and ALDH-2
activity. Biotransformation of GTN requires functionally active mitochondria and induces ROS
production and oxidative stress within this organelle, as it is inhibited by mitochondrial-targeted
antioxidants and is absent in HUVEC
0
cells. Analysis of Complex I-dependent respiration
demonstrated that its inhibition by GTN was prevented by mitochondrial-targeted antioxidants.
Furthermore, in presence of succinate (10X10
-3
mol/L), a Complex II electron donor added in order to
bypass Complex I-dependent respiration, GTN-treated cells exhibited O
2
consumption rates similar to
those of controls, thus suggesting that Complex I was affected by GTN. We propose that Complex I
becomes a target for mitochondrially-generated ROS when there is prolonged treatment with GTN in
addition to ALDH-2. Our data also suggest a role for mitochondrial-targeted antioxidants as
therapeutic tools in the control of the tolerance that accompanies chronic treatment with nitrates.

Acknowledgements: This work was supported by grants PI070091 from FIS and GVPRE/2008/307 from Conselleria de Educacin of
Generalitat Valenciana.

Key: A1
Key words: nitroglicerin, mitochondria, antioxidant

Havana Redox 2009

Antioxidants
Antioxidant Capacity Determination in Citric Juice

Domnguez Gutirrez Margarita Concepcin, Guel Silva Gabriela, Len Fernndez Olga Sonia, Victor Huerta
Espinosa, Herrera Martnez Luis Ral, Zarate Maldonado Brenda ngela, Laura Mndez Ovando, Cindy
Bandala Rodrguez, Eduardo Gmez

Benemrita Universidad Autnoma de Puebla, Puebla, Mxico

Oxidation of biological molecules and tissues is induced by oxygen and free radicals. Great part of
the antioxidant capacity of the fruits and vegetables comes from phytonutrients like vitamin C, vitamin
E, -carotene and plant polyphenols, which have a protective effect in prevention of degenerative
processes of carcinogenic and cardiovascular diseases. The aim of this work was to determine the
scavenger and reducing capacity of antioxidant in natural citric juices. Experimental design was
cross-sectional study, citric juices studied were: Orange, Mandarin and a mix Mandarin-Orange.
Juices were extracted, packaged and cooled in amber color bottles, samples were held during 48
hours before carrying out determinations. Scavenger capacity was determined using 2,2-difenil-1-
picrilhidracilo (DPPH) reagent, first reading of optical density (DO1) was done after 30 seconds being
this the value taken as control, second reading of optical density (DO2) was done after 30 min;
reducing capacity was done by FRAP method, equipments used for both tests was a
spectrophotometer 6405UV/vis Jenway. Scavenger capacity of 84.8% was observed for mandarin
juice > Mandarin orange (78%) > orange (59.9%). Reactive oxygen species reported for mandarin
juice (870.5M) > mandarin-orange (865.5M) > orange (805.5M). Therefore it is concluded that
mandarin juice and orange-mandarin mix were those with higher antioxidant capacity.

Key: A2
Key words: citric juice, DPPH, FRAP, antioxidants


Havana Redox 2009

Antioxidants
Total Anthocyanins and Total Phenolics of Capulin (Prunnus capuli) and
Their Antioxidant Properties

VM. Huerta-Espinosa, Santacruz, VC, Santacruz VV, Guel Silva G

Facultad de Ingeniera Qumica, Colegio de Ingeniera de Alimentos, Benemrita Universidad Autnoma de Puebla, Puebla, Mxico

Capulin is an edible fruit known as Mexican black cherry it grows from a tree. The beneficial effects
from fruits and vegetables have been ascribed to natural antioxidants such as anthocyanins and
polyphenolics. Recent studies show a strong association between fruit and vegetable consumption
and reduced risk of several degenerative diseases such as cancer, cardiovascular disease, and
stroke, which are caused by oxidative stress. Specifically, capulin contain natural chemopreventive
chemicals its consumption has been reported to alleviate cough and respiratory diseases. Total
anthocyanins, total phenolics, and the antioxidant activities of Capulin (Prunus capuli.) were
determined. Total anthocyanin pigment content of the aqueous extracts was determined using the
pH-differential method described by Giusti and Wrolstad. Total phenolic content was determined
using the modified Folin-Ciocalteu procedure described by Singleton and Ross. Antioxidant activities
were determined by ORAC and FRAP assays. Capulin edible portion reported antocyanin content of
31.2+2.7 (mg cyn-3-glu/100g fw), total phenolics 2.43+0.14 (mgGAE/g fw), ORAC value was 13.10 +
1.18 and FRAP 14.31 + 0.28 micrograms TE/g fw. It is concluded that antioxidant activities were
highly correlated with total phenolics. In addition, Capulin may be potential source for natural
colorants, nutraceuticals, and natural antioxidants.

Key: A3
Key words: Prunnus capuli, antioxidants, FRAP, ORAC

Havana Redox 2009

Antioxidants
Antioxidant Activity of Loquat (Eriobotrya japonica)

V M Huerta Espinosa, V Santacruz Vazquez, C Santacruz Vazquez, G Guel Silva


Loquat is a golden pear-shaped fruit with a delicate sweet-sour flavor, it averages 2.5 cm diameter. It
is native of Asia but it grows in Mexico, south US, central and South America. Loquat contain
significant amounts of secondary plant metabolites, including carotenoids, flavonols, anthocyanins,
and procyanidins. These minor dietary compounds have been postulated to play a key role in
humans as antioxidants, by preventing reactions produced by oxygen and nitrogen reactive species
during the progression of different human pathologies. Aim of this works was to determine antioxidant
activity, Randox (Randox Laboratories Ltd.) spectrophotometric kit was used to determine TAS (Total
Antioxidant Status). Trolox (Sigma) was used as a reference antioxidant, antioxidant activity was
expressed as Trolox equivalents. Antioxidant Activity was 4.2 mmol TE/kg, this result is consistent
with those found for Nectarines and Peaches. It is cloncluded that it is recommendable to incorporate
loquat in the diet to benefit of his high antioxidant activity.

Key: A4
Key words: Eriobotrya japonica, Antioxidant, Total Antioxidant Status


Havana Redox 2009

Antioxidants
Neuroprotective Effect of Aqueous Extract of Yellow Maca in Newly
Weaned Male Rat

R. Or
a
, S.Surez
a,
L.Rojas
b
, R. Valdivieso
a
I. Arnao
a
.

a
Centro de Investigacin de Bioqumica y Nutricin Facultad de Medicina
b
Facultad de Farmacia y Bioqumica; UNMSM. Av. Grau
cuadra 7 s/n, Lima 1, Per.

The Lepidium meyenii Walp (maca) is a high altitude plant from the Peruvian Andes, is traditionally used for its
nutritional value and its energizing properties, a study of neuroprotective property is presented. Objective: To
evaluate the neuroprotective effect of the aqueous extract of yellow maca in newly weaned male rats. Material
and Methods: There were four groups of newly weaned male rats: (I) control, (II) 0.4 mg maca / g weight (III)
0.8 mg maca / g weight and (IV) 1.2 mg maca / g weight. The aqueous extract was administered by orogastric
canule. It is performed a control of the weights. It was applied the test of spatial learning of Morris after 15
days of starting treatment for 4 consecutive days, the last day the test was performed 3 times.The animals
were sacrificed by decapitation and were prepared brain homogenized to 10% for the determination of the
activity of seudocolinesterasa and levels of malondialdehyde (MDA). Results: There was no significant
difference in weight gain. In applying the test of Morris, the time in seconds to reach the platform were
sequentially as follows: (I) 80, 41, 42.6 and 11.34; (II) 74.8, 28.3, 32.8 and 10.41 (III) 62 21.3, 26.8 and 8.6 (IV)
45, 15.8, 15.0 and 7.2. The enzyme activity expressed as U / g brain tissue were 10-3 x (I) 5.5, (II) 4.1 (III) and
7.5 (IV) 5.1. Both parameters show no significant differences. The levels of MDA-TBARS showed significant
differences in groups III and IV with respect to Group I. Conclusion: The administration of aqueous extract of
yellow maca has neuroprotective effect on newly weaned rats.

Key: A5
Key words: Lepidium meyenii Walp, neurobehavior, TBARS

Havana Redox 2009

Antioxidants
Antioxidants and Atherosclerosis: Effect of Hydroalcoholic Extracts of Plants
for Medical Uses on Key Processes of the Atherogenesis

O. Lara
a,b
, A. Gil
a
, R. Ramrez
a,b
, J. Londoo
a,b

a
Bioactive Substances Research Group -
b
immunomodulation Group. University of Antioquia, (a) School of Pharmaceutical Chemistry
(b) School of Medicine, University Research Center SIU, Medellin Colombia

In Latin America and the Caribbean cardiovascular diseases represents 31% of all causes of death, being
Atherosclerosis the main cause of heart disease and cerebrovascular accidents. This chronic disease of
immune-inflammatory type has been related to the activation of macrophages by oxidized low density
lipoprotein (oxLDL) and the subsequent production of unbalanced pro-inflammatory substances such as
cytokines, chemotactic factors and reactive oxygen species (oxidative stress). Epidemiological data have
shown that antioxidant rich diets are associated with a reduced risk of cardiovascular disease. In this study,
the antioxidant capacity of hydroalcoholic extracts from seven medicinal and aromatic plants used in traditional
medicine: Rosmarinus officinalis, Calendula officinalis, Hypericum perforatum, Persea americana, Junglans
regia, Eucaliptus globulus and Verbena officinalis, were evaluated. The total content of phenolic compounds
were determined using the Folin Ciocalteaus method, the protection of human LDL from oxidation was
assessed by the method TBARS and the ability to modulation of oxidative stress induced by oxLDL in murine
macrophages derived from bone marrow was measured using flow cytometry. Additionally, the interaction
oxLDL-macrophage and the ability of extracts to inhibit the uptake of oxLDL after being labeled with the DiI
fluorophore (oxLDL* DiI) was evaluated by flow cytometry and fluorescent microscopy. Data analysis revealed
that Rosmarinus officinalis despite of not having the highest amount of phenolic compounds, showed the
maximum protective effect against LDL oxidation and also reduced oxidative stress at the lowest concentration
tested (10g/mL; p <0.005); however, its ability to reduce the uptake of oxLDL was not as high as the showed
by Junglans regia, Eucalyptus globulus, Hypericum perforatum and Persea Americana extracts. Even more,
Persea Americana was able to protect LDL from oxidation and to inhibit the uptake of oxLDL more efficiently
that Verbena officinalis (p <0.005) and Calendula officinalis (p <0.005), which showed the lowest levels of
antioxidant activity, inhibition of oxLDL uptake by murine macrophages and total content of phenolic
compounds.

Key: A6
Key words: medicinal plants, antioxidant, atherogenesis


Havana Redox 2009

Antioxidants
Vitilvenz

Natural Alternative Therapy in Vitiligo

Janeth E. Rojas Urdaneta
a
, Nubia Fernndez Hernndez
b
b

a
Jerico c.a. Venezuela.
b
Centro de Estudios para las Investigaciones y Evaluaciones Biolgicas. Food Science and Pharmacy College, University of Havana,
Cuba. UH

Oxidative stress has been proposed as a part of the pathological mechanisms of melanocyte degradation in
vitiligo. It has been demonstrated a redox disruption in affected areas. New therapeutically approach based on
these facts has been proposed recently. A novel combination of natural extracts (Cucumis melo, Citrus
aurantifolia and Pimenta racemosa) named Vitilvenz
, has been tested in in vitro assays, basically in the

inhibition of phospholipids oxidation and related assays on redox mediator, involving in vitiligo
physiopathology. The sample showed an antioxidant effect when assayed on spontaneous lipid peroxidation
test in phospholipids. In addition the extract acted as a scavenger of H2O2, and free radicals, in particular
hydroxyl radical by a mechanisms involving iron quenching. A photoprotective effect of the sample has been
also demonstrated. In clinical assay the application of a cream formula bases on the above mentioned natural
extracts, in leukodermic areas of patients of vitiligo was studied. A clinical, randomized, double blind study
was carried out in 50 male and 50 female patients with stable vulgar vitiligo. Patients were distributed in five
groups as follows: Group 1 (Vitilvenz
): Vitilvenz
cream, oral administration of antioxidants and

phenylalanine. Group 2 (Placebo): placebo cream, oral administration of antioxidants and phenylalanine.
Group 3 (AF): oral administration of antioxidants and phenylalanine. Group 4 (Placebo): placebo cream.
Group 5 (Vitilvenz

A): oral antioxidant and Vitivenz
. Clinical end points: area of newly formed pigment every

30 days, during five months; and the presence of melanocytes in the histological study, at the beginning and
at the end of treatment were followed. The best clinical results was reached in Group 1, Vitivenz

plus oral
administration of antioxidants and phenylalanine (p < 0.001); followed by Group 5 that only received the
topical treatment with Vitivenz
. The clinical and histological responses of these two groups (1 and 5) were
significantly different to the rest of the groups. We concluded that melanocytes in these patients could be in a
dysfunctional state, product of the formation of free radicals that cause cellular and mitochondrial toxicity; and
that these free radicals are removed by the active components of Vitilvenz
, turning melanocytes functional

and producing melanin in the achromic area of the vitiligo. In summary, our results shown that the modification
of melanocyte redox environment may explain, at least in part, the favorable clinical effects of this natural
combination in vitiligo.

Key: A7
Key words: Vitilvenz, antioxidant, vitiligo

Havana Redox 2009

Antioxidants
Evaluation of the Antioxidant Potential of Banana (Musa Cavendish,
Musaceae) Peel Hydroethanol Crude Extract in a Wound Healing Model

A Pereira
a
, FSL Catanhede
a
, SJF Garcia
a
, KB Felipe
a
, TMF Gnther
a
, EA Ferreira
a
, MR Kviecinski
a
, AC
Vieira
a
, SRS Ferreira
b
, Correia JFG
a,
R Geremias
c
, D Wilhelm Filho
a
, RC Pedrosa
a
.

a
b
Depto. de Engenharia Qumica e Engenharia de Alimentos. Universidade Federal de Santa Catarina, Cidade
Universitria, CEP 88040-900, Florianpolis, Brazil.
c
Universidade do Extremo Sul Catarinense, Av. Universitria, CEP 88806-000,
Cricima, Brazil.

In recent years, oxidative stress and free radicals have been implicated in wound healing. In the Brazilian
popular medicine the banana peel has a history of utility for burn healing. A flavonoid galocatechin was
identified in the banana species Musa cavendish (Musaceae) and it was found to be a strong antioxidant peel
constituent
1,2
. The present study aims to evaluate the wound healing action mechanism of the hydroethanol
crude extract of Musa cavendish banana peel (MCE) considering its antioxidant potential. The antioxidant
activity was evaluated by the following determinations: contents of GSH
3
, protein carbonyls (CP)
4
and lipid
peroxydation (LP)
5
,and catalase (CAT)
6
and glutathione peroxidase (GPx)
7
activities in the skin. Balb/c mice
(weight 202g, n=54) were divided into two groups: control group (CG), which did not receive any treatment,
and the treated group (TG), which received topically MCE (400mg/kg per day) during twelve days. Results
were expressed by means and standard deviation and were analyzed using one-way ANOVA and Tukey-
Kramer test. A p-value <0.05 was considered to be statistically significant. In the dead space wound model,
the GSH content increased significantly on days 3, 9 and 12 (CG: 0.460.18 and TG: 1.670.62; CG:
1.540.40 and TG: 2.220.70; CG: 0.630.21 and TG: 4.370.95 mol.mg protein
-1
, respectively). CP level
was reduced significantly on day 3 (CG: 0.61 0.02 and TG: 1.730.42 nmol.mg protein
-1
) and also on day 9
(CG: 5.671.86 and TG: 0.730.25 nmol.mg protein
-1
). It was observed protection against lipid peroxidation in
the animals treated until day 3 (CG: 314.9155.83 and TG: 151.1259.44 nmol.mg protein
-1
). Along the next
days (6, 9 and 12) TG showed LP normalization tendency and the basal level was achieved in the wound
healing process conclusion. CAT activity returned to the basal level when the animals were treated. CAT
showed significantly difference at the days 6 and 12 (CG: 1.610.29 and TG: 1.020.31; CG: 1.760.16 and
TG: 0.940.17 mmol.min
-1
.mg protein
-1
, respectively). GPx activity was reduced extremely during all the
treatment period (day 6 CG: 8.320.06 and TG: 1.340.96; day 9 CG: 8.781.54 and TG: 0.400.09; day 12
CG: 9.971.49 and TG: 0.210.07 mol.min
-1
.mg protein
-1
). These findings indicate that the antioxidant
potential of MCE can be proposed as a possible wound healing action mechanism.
References: 1. SOMEYA, S.; YOSHIKI, Y.; OKUBO, K. Food Chemistry, v. 79, pp. 351354, 2002. 2. KONDO, K.; KURIHARA, M.;
MIYATA, N.; SUZUKI, T.; TOYODA, M. Archives of Biochemistry and Biophysics, v. 362, pp.7986, 1999. 3. BEUTLER, E.;
DURAN, O.; KELLY, B. M. 4. LEVINE, R. L.; GARLAND, D.; OLIVER, C.N.; AMICI, A. CLIMENT, I.; LENZ, A.G.; AHN, B.W.;
SHALTIEL, S.; STADTMAN, E. R. Methods Enzymology, v. 186, pp. 464-478, 1990. 5. HERMES-LIMA, M.; WILLMORE, W. G.;
STOREY, K. B. Free Radical Biology & Medicine, v. 19, n. 3, pp. 271-280, 1995. 6. AEBI, H. Methods in Enzymology, v. 105, pp.
121-126, 1984. 7. FLOHE, L.; GUNZLER, W.A.Methods in Enzymology, v. 105, pp. 114-121, 198

Key: A8
Key words: Musa Cavendish, antioxidant, wound healing

Havana Redox 2009

Antioxidants
Hepatoprotective Effect of 2,4,6-trihydroxiacetophenone Against Oxidative
Stress Induced by Carbon Tetrachloride in Mice

Eduardo A. Ferreira
a
, Jussara M. Rebello
a
, Eliana F. Gris
a
, Karina B. Felipe
a
, Danilo Wilhelm Filho
b
, Rosendo
A. Yunes
c
, Rozangela C. Pedrosa
a

a
b
Depto. de Ecologia e Zoologia ,
c
Depto. de Qumica Universidade Federal de Santa Catarina, Cidade
Universitria, CEP 88040-900, Florianpolis, Brazil.

This study investigated the protective effects of phloroacetophenone (2,4,6-trihydroxiacetophenone, THA), an
acetophenone derived from Curcuma comosa, which also can be produced by organic synthesis, on carbon
tetrachloride-induced hepatotoxicity, as well as the possible mechanisms involved in such protection in mice.
This compound exhibited in vitro antioxidant effects on FeCl2-ascorbate induced lipid peroxidation in mouse
liver homogenate, and also exhibited hydroxyl and superoxide radical scavenging activity. In vivo assay: a
control group received saline once a day during 7 days and on the 8
th
day received only CCl
4
vehicle (Control);
another group (CCl4), received saline once a day for 7 days and on the 8th day received CCl4 (0.5 mL/kg body
weight, injected intraperitoneally); and finally the THA group, which was pre-treated with THA (6.0 mg/kg body
weight, administered orally by gavage) once a day for 7 days. Pre-treatment with THA prior to the
administration of carbon tetrachloride significantly prevented the elevation of serum enzymatic activities of
alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase. In addition, pre-treatment
with THA significantly prevented the increase of hepatic lipoperoxidation measured by TBARS levels and also
protected the hepatocytes against protein carbonylation and oxidative DNA damage. Consistent with these
observations, THA pre-treatment normalized the activities of antioxidant enzymes such as catalase (CAT) and
superoxide dismutase (SOD). These results suggest that the protective effects of THA pre-treatment against
carbon tetrachloride-induced hepatotoxicity are due to decreased CCl4-mediated oxidative damage resulting
from the inherent antioxidant properties of THA.

Key: A9
Key words: 2,4,6-trihydroxiacetophenone, antioxidant, CCl
4
, hepatotoxicity


Havana Redox 2009


Havana Redox 2009

Antioxidants
Anthraquinones Extracts From Different Organs of Morinda royoc L. In Vitro
Plants and Their Antioxidant Properties.

Janetsy Borroto
a
, Josep Coll
b
, Maribel Rivas
a
, Maria Blanco
a
, Oscar Concepcin
a
, Yudelsy A. Tandrn
b
,
Martha Hernndez
a
, Reinaldo Trujillo
a
.

a
Laboratorio de Ingeniera Metablica. Centro de Bioplantas. UNICA. Carretera a Morn km. 9. CP 69450. Ciego de vila. Cuba e-
mail: jborroto@bioplantas.cu

b
Instituto de Investigaciones qumicas y Ambientales de Barcelona Josep Pascual Vila, Consejo Superior de Investigaciones
Cientficas, Jordi Girona 18-26, 08034-Barcelona, Spain.

Plant antioxidants are very attractive as a natural alternative to synthetic compounds. Natural extracts used in
food, pharmaceutical and cosmetic have been more valorized by customers. This study compared
Anthraquinones (AQs) contents and antioxidant activity in different organs of Morinda royoc L. in vitro plants,
focus on root culture AQs and identification by H
1
-NMR. The AQs constitute an important group of natural
products. These compounds have been reported to exhibit some interesting in vitro biological activities: anti-
microbial, anti-fungal, anti-malarial, anti-oxidant and anti-mutagenic. The analyzed organs exhibited different
total AQs contents (0.33-4.5 mg.g
-1
of fresh weight), which do not correspond to the anti-oxidative activity
measured. This is probably due to the presence of different phenolics compounds in the samples. The
phenolics compounds can presents anti-oxidative activities which involves various mechanisms to avoid the
oxidation process.The highest AQ content was recorded in root culture showing significant statistical difference
compared with other organs. The stem extracts displayed the highest anti-oxidant activity with the lowest
malondialdehide and other aldehide value. The antioxidant activity found in roots culture can be due to the
AQs compounds present in the extract. Seven AQs were isolated from root culture with sufficient amount to be
characterized. The AQs were isolated from the dichloromethane extract and purified by silica gel column using
a reverse phase high performance liquid chromatography coupled with photo-diode array detection (HPLC-
DAD). The structures of the AQs were established based on spectral studies (H
1
-NMR). Seven AQs were
identified. One of them namely Rubiadin constitutes the first report of their occurrence from Morinda royoc L.
and six another known AQs: Nordamnacanthal. Damnacanthal, Morindone, Soranjidiol, Lucidina and Rubiadin-
1-methylether.

Key: A10
Key words: Antioxidant activity; root culture; Rubiaceae, secondary metabolism


Havana Redox 2009

Antioxidants
Antioxidant and Antigenotoxic Evaluation of DI-31, a Brassinosteroid
Analogue

I Gonzlez-Blanco
a
, Y Prez Torres
b
, R Gonzlez
b
, J Dranguet-Vaillant
b
, F Coll-Manchado
c
and S Daz-Llera
a

a
Department of Pharmacology and Toxicology, Pharmacy and Food Sciences College, 25
th
ivongb@ifal.uh.cu, sdllera@infomed.sld.cu.
b
Centre for Research and Biological Evaluations (CEIEB),. 23
rd
c
Faculty of Chemistry, Havana University, San Lzaro and L, 10400, Havana, Cuba

DI-31 is a brassinosteroid analogue phytohormone with a high plant growth-promoting and anti-stress activity
1
.
It has remarkable effects on cell elongation and division and is employed to improve crop yield and crop
quality. The aim of the study was to determine the antioxidant effect in vitro and the antigenotoxic activity in
vivo. The antioxidant effect was evaluated using biochemical assays which detect a) the reactive oxygen
species scavenging capacity (DPPH, H2O2, HO
) and b) the antioxidant protecting mechanisms to

biomolecules (proteins, lipids and DNA), and to intact cells with the Comet assay in human leukocytes
2
. The
antigenotoxic activity was studied with the mouse bone marrow micronuclei (MN) assay. Six concentrations of
DI-31 (0.042-0.17g/mL) were used for the in vitro tests. Appropriate control chemicals were employed in
each assay. Male Balb/C mice (5/group) were pretreated with four doses of DI-31 (0.03-30g/kg) by o.g.
Ciclophosphamide (CP, 40 mg/kg b.w., i.p.) and Bleomycin (BLM, 60 mg/kg b.w., i.p.) were used as mutagens
and Vitamin E (100 mg/kg b.w., o.g. ) as reference control. DI-31 (0.10 and 0.13 g/mL) moderately
scavenged the HO
. A similar protecting effect of DI-31(0.10-0.17 g/mL) was observed with respect to the
oxidative damage induced to lipids and DNA. 0.10 and 0.13 g/mL of DI-31 significantly protected HO

challenged proteins with 95% inhibition of carbonyl groups. In the Comet assay, DI-31 (0.10g/mL) was 30%
less active than Vitamin E (10
-5
M) regarding to protection against oxidative damage produced by H
2
O
2
of
human leukocytes DNA. Micronuclei frequency decreased in mouse bone marrow for both mutagens (BLM
and CP). The group treated with DI-31 (3 mg/kg b.w.) + BLM showed a significant 60% reduction of MN
frequency with respect to the BLM control group. Four doses of DI-31 (0.03; 0.3; 3 and 30 mg/kg b.w.)
significantly reduced the genotoxic effect induced by CP. The percentages of inhibition of the MN frequency
ranged from 55.7% with the lowest active dose to 71.6% with the highest dose. The in vitro tests make
possible to conclude that DI-31 is a moderate antioxidant which slightly protects lipids and considerably
prevent oxidative damage to protein and DNA. In vivo, DI-31 has an important antigenotoxic effect against two
potent mutagens BLM and CP.
References:
1. Sakurai, A. Yokota, G. et al. (2000) Brassinosteroids: Steroidal Plant Hormones. Academic Press. California, USA.
2. Collins, A.R. (2002) The Comet Assay, from Methods in Molecular Biology 203, In Situ Detection of DNA Damage: Methods and
Protocols, Human Press Inc., Totowa, NJ.

Key: A11
Key words: antigenotoxic; antioxidant; brassinosteroid

Havana Redox 2009

Antioxidants
Antioxidant Protection of Vascular Endothelium in Oxidative Damage
Models to Cultured Endothelial Cells.

Pereira N, Garca JC, Rivern G, Gutierrez R, Pupo JB, Pandolfi A.

*National Center of Medical Genetics. Higher Institute of Medical Sciences of Havana.
Address: 146 No. 3102. Playa, PC 10600, Havana City, Cuba. Phone: (53) (7) (208) 9991 - 9999 ext.1093.

Introduction: Cardiovascular disease is the first cause of death all over the world. The importance of the
vascular endothelium in the physiopathology and the essential role of the oxidative stress in the molecular
mechanisms have been demonstrated. Objective: Evaluate the response of cultured endothelial cells to the
oxidative damage induced in three different models and evaluate the possible protection using antioxidants.
Materials and Methods: Endothelial cells from H5V murine cell line were cultured under oxidative stress
condition using: isoprostane 8-iso-PGF2
H2O2 and the copper (II) complex of 1, 10-phenanthroline. Ascorbic
acid, coenzyme Q10, bovine serum albumin (BSA) and the heat shock protein -crystalline were used to
evaluate possible antioxidant protection. Cellular survival was tested by MTT assay and the type of cell death
was evaluated by Tunel, ELISA and double stain fluorescence microscopy. Comet assay was used to evaluate
DNA protection in H
2
O
2
- treated cells. Results and Conclusions. Cells survival decreased in the three models
in dose and time dependent way. The treatment with antioxidants increased cell survival, except when we
used ascorbic acid in the presence of H
2
O
2
. Necrosis was the kind of cell death induced in our experimental
conditions. Crystalline treatment increased cell survival significantly compared with BSA, a protein with
antioxidant properties known. Oxidative damage induces cell death of the endothelial cells in culture.
Antioxidants can attenuate the effect of oxidative damage in cell culture condition. Our results showed that the
heat shock protein -crystalline could be considered as endothelial protector according to its antioxidant
properties join to the chaperone ability.

Key: A12
Key words: endothelium; antioxidant; -crystalline

Havana Redox 2009

Antioxidants
Content of Redox-Active Compounds in Foods Consumed in Macrobiotic
Diet

J L Rodrguez
a
, D Gonzlez
b
, V Huerta
c
, D Castro
b

a
Instituto de Investigaciones para la Industria Alimenticia, Carretera al Guatao km 3, C.P. 19200, La Habana, Cuba.
b
Instituto Finlay, La Habana, Cuba
c

Macrobiotics is predominantly vegetarian, whole foods diet that has gained popularity in Cuba due to
individuals' cases reports that have been recovered from diabetes. For this reason, the main objective of this
study was to estimate antioxidant capacity of foods regularly consumed in a diet established and applied by a
group of specialists from Institute Finlay. Sampling of 19 foods was performed as they are consumed being
aware of typical culinary practices of this diet, antioxidant capacity was determined using FRAP test (Ferric
Reducing Antioxidant Power) of Benzie and Strain, and results were expressed as mg ascorbic acid equivalent
per 100 g of sample. Observed values for vegetable group (n = 12) were between 6 and 234 mg of ascorbic
acid equivalent, being parsley and chicory among those with higher values. For whole cereal group (n = 3)
antioxidant capacity varied between 7 and 200 mg and the best results were for rice and barley. Finally,
legume group (n = 4) values were between 36 and 1150 mg, corresponding to black and red beans the greater
values.

Key: A13
Key words: macrobiotic; antioxidant


Havana Redox 2009

Antioxidants

Determination of Antioxidant Capability of Drinks of Jamaica and Tamarind

Danae Prez Santana

Instituto de Farmacia y Alimentos .Universidad de la Habana. E.mail: danaeps@ifal.uh.cu

The antioxidant capacity and the relation with quantity to phenolic compounds of natural Jamaica flower
(Hibiscus sabdariffa), tamarind (Tamarindus indica L.) and its industrial forms of drinks, juices, tea were
evaluated. The dry flower drink of the Jamaica presented the most antioxidant capacity (32 meq / L) and the
most quantity of total phenolic components (1397 mg / L). The pulp and juices of tamarind possess
perspectives being funtional drinks with antioxidative potentials, although they didnt reach the potential that
the Jamaica drink and tea.

Key: A14
Key words: Hibiscus sabdariffa, Tamarindus indica, antioxidant capability, phenolic compounds.


Havana Redox 2009

Antioxidants
Antioxidant and Wound Healing Effect of Rhizophora mangle (L.) in an
Aseptic Wounds Model in Rats.

Janet Snchez,
a
Roberto Faure,
a
Gregorio Martnez,
b
Octavio Fernndez,
a
Ernesto Vega
a
and Evangelina
Marrero
a

a
Dep. Chemistry, Pharmacology and Toxicology, National Centre for Animal and Plant Health, CENSA, Aptdo.#10, San Jos de Las
Lajas, La Habana, Cuba.
b
Pharmacy and Food Sciences Institute, University of Havana, Cuba. E-mail: jsanchez@censa.edu.cu

Rhizophora mangle (L.) bark aqueous extract and its major component, high molecular weight polyphenols
fraction, were evaluated for antioxidant and wound healing activities in a rat model of aseptic wounds. The
wound area (mm
2
) was used as parameter of wound healing and was measured with Imagen Digital
Processing System MADIP 3.0 Lab Lapdis from Carlos J. Finlay Hospital, at 7 days after done the wounds.
Differents markers were evaluated in the new tissue in order to know the redox balance during wound healing
process at 0, 3, 7 and 14 days after to start the treatment: Dismutase Superoxide (SOD), Catalase (CAT),
reduced Glutahione (GSH) and malonildialdehyde (MDA). Results showed that the wound area was reduced
with the extract and its fraction treatment compare to control group (p< 0.05). Also the activity of antioxidants
enzymes SOD and CAT, and GSH concentrations were increased in presence of the extract, its fraction and
positive control compared to control group (p< 0.05); while the MDA concentrations was decreased in these
groups (p< 0.05). This study demonstrated that Rhizophora mangle (L.) modified the redox balance, increasing
the antioxidant biomarkers and decreasing prooxidant indicators in the wound area and in this way the
antioxidant properties of extract and its fraction contributed to improvement wound healing process.

Key: A15
Key words: Rhizophora mangle (L.), antioxidant, wound healing


Havana Redox 2009

Antioxidants
Fruits and Vegetables Ingested by Hypertensive Women: Relationship of
The Lipidic Profile and The Antioxidant Total Capacity in Serum.

L Ledesma, A Gutirrez, I Garca

Food Science and Pharmacy College, University of Havana, Cuba

The relation of the biomarkers of lipidic profile and the antioxidant total capacity in serum (TAS), was evaluated
in hypertensive women, with different diet ingestion of fruits and vegetables. According to the dietetic evaluation
of the hypertensive women, they were classified in four groups, based in the growing consume of fruits and
vegetables. Significant levels of the media values were founded for the TAS and the HDL cholesterol, in relation
to the increment of the consume of fruits and vegetables. Mathematical expressions were developed finally to
show the direct relationship between the uptake of fruits and vegetables with the TAS and HDL, and an inverse
relation with the LDL.

Key: A16
Key words: hypertension, lipids, antioxidant, fruits, vegetables


Havana Redox 2009

Antioxidants
In vitro Antioxidant Activity and Chemical Composition of Halimeda monile
Seaweed Aqueous Extract

Batista AE
a
, Costa A
a
, Zaldvar C
a
, Olivares S
b
, M Prez
c
, Mancini J
d
, Vidal A
a

a
- Department of Biochemistry, Faculty of Biology, University of Havana, Cuba
b
- Laboratory of Environmental Research, Institute of Technologies and Applies Sciences, Cuba
c
- Center for Natural Products Studies, Faculty of Chemistry, University of Havana, Cuba
d
- Department of Food and Experimental Nutrition, Faculty of Pharmacy, University of Sao Paulo, Brazil

The quest for antioxidant compounds extracted from natural sources is a field with expanding possibilities, due
to the role of free radicals in diverse pathologies. In this work the antioxidant activity of Halimeda monile
seaweed aqueous extract was studied in different in vitro models: spontaneous lipid peroxidation of rat brain
homogenates, DPPH
.
radical scavenging activity and reducing power. A pharmacognostic and chemical study
of the seaweed was also carried out, and the content of total phenolics and minerals was determined. H.
monile aqueous extract exhibited a dose-dependent effect against lipoperoxidation (IC
50
=1.09 0.07 mg/mL)
and for DPPH
.
radical scavenging (IC50 of 0.4 0.007 mg/mL) and also showed a significant reducing power.
The pharmacognostic study was positive for triterpenoids, but no alkaloids, saponins or flavonoids were found.
Total polyphenols a content was 26.4 mg of salicylic acid equivalents/ g of lyophilized. Mineral analysis
indicated presence of Cu (8.1 0.7 g/g of dry powder), Ni (18.7 1.2 g/g of dry powder) and Zn (< 0.8 g/g
of dry powder). The antioxidant activity of Halimeda monile could be related to the presence of phenolic
compounds and terpenoids that could contribute to the reducing power and free radical scavenging of the
extract.

Key: A17
Key words: Halimeda monile, antioxidant

Havana Redox 2009

Antioxidants
Protective Effect of Bidens pilosa L Extract in Hepatotoxicity Induced by
Paracetamol.

Rayza Mndez Triana, Mara BoffIill Crdenas, Emilio E. Monteagudo Jimnez, Maykel Prez Machn,
Omaida Lpez Bernal.

Medical College of Villa Clara. Clinical Trials Center. Acueduct Rd. and Circunvalation. Santa Clara, Villa Clara, Cuba.
maykelpm@iscm.vcl.sld.cu

Bidens pilosa L, known as romerillo blanco, is widely used by the cuban population. It is chemically composed
by flavonoids (quercetin) with proved antioxidant properties. A preclinical study was carried out to evaluate the
hepatoprotective effect of Bidens pilosa L in the toxicity induced by paracetamol. Male adults MNRI mice were
used to evaluate the effect of oral administration of three-level of dose (150, 300 y 600 mg/kg weight) of
Bidens pilosa L extract, 30 min before and 2 hours after the hepatotoxicity induced by paracetamol (600
mg/kg, oral, unique dose) establishing as markers the evaluation of: clinical signals, enzymatic activity of
alkaline fosfatase, alanine aminotransferase (ALT) and aspartate aminotransferase (AST); and histological
changes produced by hepatocites. Abnormalities in the behavior of animals were not evidenced during the
trials. The analysis of plasmatic concentrations of the tree enzymes in the groups treated with the plant
showed high significant statistical differences with respect to control group treated with paracetamol only.
Values within the reference values range for the specimen were obtained. Anatomopathological results confirm
the cellular protection, either macroscopic or microscopically, in the livers of the animals treated with the plant
extract. To sum up, the administration of Bidens pilosa L extract showed a protective effect against the liver
damage induced by paracetamol, with no relation between doses and pharmacological action.

Key: A18
Key words: Bidens pilosa, hepathoprotection, paracethamol, toxicity

Havana Redox 2009

Antioxidants
Antioxidant Activity and Atheroprotective Effect of an Aqueous Extract of the
Marine Seaweed Halimeda incrassata

Claudina Zaldvar
a
, Ariadna Costa
a
, Ana Elsa Batista
a
, Andriy Samokhin
b
, Saadath Hashamiyan
b
, Boram
Nho
b
, Adyari Fallarero
c
, Alexis Vidal
a
, Dieter Brmme
b

a
Faculty of Biology, University of Havana, Cuba
b
Faculty of Dentistry and Center for Blood Research, University of British Columbia, Canada

c
Department of Biochemistry and Pharmacy, Abo Akademi University, Finland

It is considered that targeting oxidative modifications can retard atherosclerosis progression as oxidative stress
is critically involved in atherosclerotic lesion advancement. In previous studies it was found that H. incrassata
(Hi) had an inhibitory effect on in vitro lipoperoxidation. This work had as objectives to continue the study of the
antioxidant activity of H. incrassata and to assess its atheroprotective activity during advancement of
experimental atherosclerosis. In vitro atheroprotective effect was evaluated by the inhibitory action of Hi on low
density lipoprotein (LDL) oxidation mediated by cupper ions, effect probably related to the capacity of the
extract to scavenge
OH

(IC
50
= 1,64 mg/mL) and O
2
(IC
50
= 0.50 mg/mL) radicals. To evaluate the in vivo
atheroprotective capacity of Hi apo E-/- mice were used. At six weeks of age animals were divided into 4
groups that received for 8 or 16 weeks a hypercholesterolemic diet and the groups supplemented with
seaweed were given 550 mg lyophilized of Hi/ kg body weight/ day dissolved in drinking water. Hi
supplementation caused a reduction in lesion size and total aortic lesion area at 8 weeks and decreased lesion
severity at 8 and 16 weeks with lower macrophage infiltration. The positive effect could be related to the
antioxidant potential of the extract as we also noticed a reduction in nitrotyrosine staining in the plaques of the
animals. The results obtained suggest that supplementation with Hi could influence atherogenesis in apo E-/-
mice and represents another step in the introduction of Hi for a nutritional or phytotherapeutic application.

Key: A19
Key words: Halimedaincrassata, antioxidant, atheroprotection


Havana Redox 2009

Antioxidants
): Antioxidant Properties in
Inflammatory Models

Prez MR
a
, Rodrguez CC
a
, Martnez G
b
, Horta D
b
, Chacon A.L
a
, Fraga J.A
a

a
Laboratorios Biolgicos Farmacuticos, LABIOFAM, Ave Independencia km16 Santiago de las Vegas, C. Habana Cuba.
b
Centro
de Estudios y Evaluaciones Biolgicas, CIEB, Instituto de Farmacia y Alimentos Universidad de La Habana, Cuba. Ave 23 # 21425
e/ 214 y 222, la Coronela, La Lisa C.P. 13600 C. de la Habana Cuba.

Musa paradisiaca L is a plant of the family Musaceae. Chemical constituents of the extract have been
investigated, and some of them identified. The role of free radicals has been shown in various pathological
conditions, such as, inflammation, arthritis and others, and are remove by antioxidant defense mechanism . In
the present study we investigated the effects of the Musa paradisiaca extract on antioxidant system and on
inflammatory models in a rat subplantar oedema and ulcerative colitis induced by acetic acid. For subplantar
oedema female Wistar rats were divided into five groups: Acitan groups (54, 64, 74 mg/kg respectively p.o),
indomethacin group and control group. Oedema was induced by subplantar administration of carragenan.
Phospholipase A2 and Peroxidase levels were evaluated in sample of blood. For ulcerative colitis female Wistar
rats were divided into tree groups: Acitan group, Azulfidine group as a positive control group and acetic acid
control. Colitis was induced by intracolonic administration of 3% acetic acid. Colonic inflammation was
evaluated by colonic myeloperoxidase (MPO), SOD, CAT activity, MDA levels, macroscopical mucosa
evaluation, weight of colon, mastocites degranulation in colonic mucosa. Compared to the control group, both
Acitan and Indomethacin showed a significant antinflammatory effect of oedema subplantar (p<0.05) the ED50
estimated was 57, 54 mg/kg, the treatment with Acitan decreased the levels of PLA2 and Peroxidasa respect to
control group (P<0.05). The treatment with Acitan decreased significatively colonic concentrations of MPO and
MDA respect to control group(P<0.001), Acitan show increase of levels of SOD and CAT in the homogenate of
colon respect to control group(P<0.05). Additionally, levels of mastocites degranulation in colonic mucosa was
significantly lower in the Acitan group and Azulfidine group than in the acetic acid group (P<0.005). Acitan
could reduce inflammatory injury in rat models of subplantar oedema and ulcerative colitis at least in part by
antioxidants mechanism.

Key: A20

Havana Redox 2009

Key words: Musa paradisiaca, Acitan, antioxidant, inflammation

Havana Redox 2009

Antioxidants
Piper auritum HBK Cream: Antinflammatory Activity by Antioxidant
Mechanisms.

Rodrguez CC
a
, Prez MR
a
, Martnez G
b
and Horta D
b

a
Laboratorios Biolgicos Farmacuticos, LABIOFAM, Ave Independencia km16 Santiago de las Vegas, C. Habana Cuba.
b
Centro
de Estudios y Evaluaciones Biolgicas, CIEB, Instituto de Farmacia y Alimentos Universidad de la Habana, Cuba. Ave 23 # 21425 e/
214 y 222, la Coronela, La Lisa C.P. 13600 C. de La Habana Cuba.

In the inflammatory disease, the free radicals play an important role in the pathological process. Piper auritum
HBK is a folk medicine plant used in the treatment of the inflammation. A formulation (cream) based in a
extract of this plant is object of the present work in which we have the purpose to determinate the effect of the
cream of P. auritum HBK on inflammatory model. Rats SD weighting 180-200 g b.w. were used. Four pellets of
cotton were implanted in the abdominal region of the rats. Three experimental groups were formed: control
group (without treatment), group (cream of P. auritum) and positive control (dexametasone). The products
were applied topically twice daily during 7 days. The granulome formed were extracted and the dry weigh was
determinated for two granulomes for each animal. The others two were homogenized for biochemical study
(SOD, CAT, XO and MPO). The topical application of the cream of Piper auritum HBK in the experimental
model of implantation of cotton pellet in rats decrease the granulomatous tissue respect to the control group
(p<0.05). Additionally, the activity of myeloperoxidase enzyme was inhibited respect to the control group
(p<0.05) in the homogenate of the granulome. The enzymes of the antioxidant system, SOD and CAT, showed
an significative increase (p<0.05) respect to the control group. Otherwise, the activity of Xantine Oxidase
enzyme was depleted (p<0.05)respect to the control group. Our results evidence that the inflammatory effect
of Piper auritum HBK cream was carried out by antioxidant mechanisms.

Key: A21
Key words: Piper auritum, antioxidant, antinflammatory


Havana Redox 2009

Antioxidants
Therapeutic Effect of Mangifera indica L. in Minor Recurrent Aphthous
Stomatitis

Hetzel de la C. Lourido Prez
a
b
, Gema Prez Davison
b

a
Clnica Estomatolgica Provincial Docente Antonio Briones Montoto, Calle Sol s/n. Pinar del Ro, Cuba. E.mail:
hetzel@princesa.pri.sld.cu
b
CEIEB-Instituto de Farmacia y Alimentos, La Habana, Cuba

The aim of this study was to determine the therapeutic effect of Mangifera indica L. (Vimang) oral gel
formulation in minor Recurrent Aphthous Stomatitis. It was made a stage II clinical trial, controlled,
randomised, longitudinal and prospective study, in Antonio Briones Montoto Pinar del Ro Dental Clinic,
between March and October 2007. The total number of individuals enrolled was 35; it was conformed two
groups, one of them involves 18 individuals treated with Vimang and the other one conformed by 17
individuals treated with 0.2% Clorhexidine mouth rinse. It was included patients suffering minor Recurrent
Aphthous Stomatitis who did their participation consent. Patients were checked at 24, 48, 72 hours and later at
5
th
, 7
th
and 10
th
days. Statistic analysis was made using X2 and Mantel and Hanszelses X2, Fishers exact
probability; statistic comparison was verified at 95% of confidence. It was obtained a similar time of evolution in
both groups (seven days); in the 55.6% of patients treated with Vimang the pain disappeared at 24 hours;
the number, size and time in witch lesions appearing had not influence in a treatment response. It wasnt found
relation between demographic variables, lesion location and clinical evolution, it wasnt found any adverse
effect, and the 88.9% of the study group accepted the new product. The Vimang gel is effective in minor
Recurrent Aphthous Stomatitis treatment, specifically in pain relief.

Key: A22
Key words: Mangifera indica,Vimang, recurrent aphthous stomatitis, clinical trial


Havana Redox 2009

Antioxidants
Antioxidant Activity of Six Aromatics Plants Used in Cuba as Condiments

O Valds
a
, JL Rodrguez
a
, Y Saborit
b
, O Marante Maldonado
a
, O Zerquera
a
.

a
Instituto de Investigaciones para la Industria Alimenticia, Carretera al Guatao km 3 ,CP 19200, La Lisa, La Habana, Cuba.
b
Instituto de Farmacia y Alimentos, calle 222 e/ 25 y 230, La Coronela, CP 19200, La Lisa, La Habana, Cuba.

The aromatics herbs and condiments have been very useful for the flavoring of different beverages and food
preparations. They have been employed in traditional medicine and their essential oils are very important for
food and pharmaceutical industries. Recently, it has been attributed antioxidant properties due to phenolic
components, such as flavonoids, phenolic acids, phenolic diterpenes, etc., which are present in their
composition. In the present study, it was investigated the antioxidant activity of alcoholic extracts of six
aromatic herbs commonly used in home preparation, they were: wild oregano (Coleus amboinicus Lour.),
parsley (Petroselinum crispum M.), chive (Allium schoenoprasum L.), spearmint (Mentha spicata L.), coriander
(Coriandrum sativum L.) and basil (Ocimum basilicum L.). Antioxidant capacity was assessed by the Ferric
Reducing Antioxidant Power (FRAP) and cido 2,2-azinobis-(3-etilbenzotiazolin-6-sulfnico (ABTS) assays
respectively. The phenolic content of the herbs was also evaluated. As result of this study was found that
among the extracts analyzed, wild oregano exhibited the highest antioxidant power. The highest phenolic
content was found in basil extract. The lowest values of antioxidant capacity were showed by chive, coriander
and parsley which were those of lowest phenolic content. The antioxidants capacity values obtained for wild
oregano were comparables whith those of the Greek oregano, aromatic plant considered with a high
antioxidant capacity.

Key: A23
Key words: Coleus amboinicus, Petroselinum crispum, Allium schoenoprasum, Mentha spicata, Coriandrum
sativuml, Ocimum basilicum, antioxidant

Havana Redox 2009

Antioxidants
Absorption and Metabolism of Flavonoids and Their Metabolites

L. Rodolfo

Pharmacy and Foods Institute. University of Havana. Ave. 23 No. 21425 e/ 214 and 222. La Coronela. La Lisa

Dietary flavonoids, a class of semi-essential food components, have long been believed to exert protective
effects against many diseases, in particular cardiovascular disease , cancer and neurodegenerative disorders.
The biological effect of these polyphenols and their in vivo circulating metabolites will ultimately depend on the
extent to which they associate with cells, either by interactions at the membrane or more importantly their
uptake. This has been well supported by a myriad of studies examining potential sites and modes of action.
This work summarizes the current knowledge on the cellular uptake of flavonoids and their metabolites with
particular relevance to further intracellular metabolism and the generation of potential new bioactive forms. The
impact of membrane transporters as well as metabolic enzymes on the cellular availability of these
phytochemicals are examined.

Key: A24
Key words: flavonoids, metabolism, absorption

Havana Redox 2009

Antioxidants
Isolation of Antioxidant Compounds from Medicinal Extract of Pedilanthus
Tithymaloides

Gonzlez TL,
a
Abreu P,
b
Matthew S,
b
Costa D,
c
Segundo M,
c
Fernandes E
c

a
Departamento de Farmacia, Facultad de Ciencias Naturales, Universidad de Oriente, Santiago de Cuba; Cuba, email:
tlopez@cnt.uo.edu.cu;
b
CQFB/REQUIMTE, Faculdade de Cincias e Tecnologia da Universidade Nova de Lisboa, 2829-516 Caparica,
Portugal;
c
REQUIMTE, Departamento de Qumica-Fsica, Faculdade de Farmcia da Universidade do Porto, Rua Anibal Cunha 164,
4090-030, Porto, Portugal.

In Cuban traditional medicine, the tincture of P. tithymaloides is used as an anti-inflammatory remedy in the
treatment of stomatological affections. The active principles responsible for these medicinal properties, was
determined using a methodology of bioguided isolation, monitored by TLC autographic assay of 2,2-diphenyl-
1-picrylhydrazyl (DPPH) radical the scavenging effects of the tincture against reactive oxygen species (ROS)
and reactive nitrogen species (RNS), with the following results: HO (IC
50
= 345 77 g/mL); O
2

(IC
50
= 143
7 g/mL); HOCl (IC50 = 113 20 g/mL); ONOO
(IC50 = 44 3 g/mL); ONOO
in the presence of
NaHCO
3
(IC
50
= 82 12 g/mL); and NO (IC
50
= 54 4 g/mL). The extract of stem and leaves of P.
tithymaloides was eluted over a RP-18 column with H
2
O, H
2
O/MeOH (1:1), MeOH, and CH
2
Cl
2
, and the
corresponding fractions tested against
NO, ONOO
, and ROO
. The MeOH/H2O (1:1), and the MeOH fractions

proved to be the most active, with the following IC
50
values (g/mL):
NO (MeOH/H
2
O, 11.9 2.2; MeOH, 21.1
8.4); ONOO

(MeOH/H
2
O, 2.5 0.2; MeOH, 16.3 1.1); ONOO

in the presence of NaHCO
3
(MeOH/H
2
O,
1.9 0.3; MeOH, 5.6 1.1); ROO

(MeOH/H2O, 1157 53; MeOH, 422 39. The flavonoids kaempferol 7-O-
D-glucopyranoside-6-(3-hydroxy-3-methylglutarate), quercitrin, isoquercitrin, and the coumarin scopoletin
was isolated and identified. The results here presented indicate that these phenolic compounds constitute part
of the active principles of the plant extract responsible for its antioxidant activities.

Acknowledgements: REQUIMTE and Fundao para a Cincia e Tecnologia (Portugal).

Key: A25
Key words: Pedilanthus Tithymaloides, flavonoid, Coumarin, Antioxidant activity


Havana Redox 2009

Antioxidants
Phenylpropanoid Content in Plant of Theobroma cacao L.: Antioxidant
Activity

J. Quiones, R. Trujillo, T. Augustine, Y. Capdesuer, Y. Quirs, J. Borroto, M. Hernndez.

Bioplant Center. University of Ciego de vila. Carretera a Morn km 9 CP 69450. Ciego de vila. Cuba. Telf:(53-33)224016/225768.
Fax: (53-33)266340. e-mail: jquinones@bioplantas.cu

The Theobroma cacao L. (Cocoa) is endemic to the Amazon basin and Central America. Cocoa-derived foods
are rich in phenol and is obtained from the fermented, roasted and milled seeds of Theobroma cacao L. A
growing body of evidences indicate that secondary plant metabolites plays a critical role in human health and
may be nutritionally important. Of special interest are plant-based phenolic metabolites due to their potent
antioxidant activity and wide range of pharmacologic properties including anticancer, antioxidant, and platelet
aggregation inhibitory activity. Antioxidants are compounds that can delay or inhibit the oxidation of lipids or
other molecules by inhibiting the initiation of propagation of oxidizing chain reactions. The antioxidant activities
of phenolic compounds are mainly due to their redox properties, which can play an important role in adsorbing
and neutralizing free radicals. The aim of this work were to determinate the phenolic content in plant of
Theobroma cacao L. and the antioxidant activity of the extracts. Leaves, branches, seeds, flowers and roots of
field plant, and callus of staminodes, petals and nucelus were analyzed. The Phenolic content was
determinated with Folin-Ciocalteu reagent according to the method of Gurr et al. (1992). The procedure for
performing antioxidant activity were based on Harding and Benson (1995) and Ait Barka et al. (2000) for
malondialdehyde and aldehyde levels respectably. Statistical Package for Social Sciences (version 11.5 for
Windows, SPSS Inc.) and parametric test (One Way ANOVA-Tukey (p>0.05)) was utilized for statistic
analysis. These studies showed that leaves and roots have more phenolic content than the other organs
evaluated, and a higher antioxidant activity was associated with these phenolic content.

Key: A26
Key words: phenylpropanoid, Theobroma cacao, Antioxidant activity


Havana Redox 2009

Antioxidants
Mangifera indica L Extract (Vimang) and Its Main Polyphenol Mangiferin
Prevent Mitochondrial Oxidative Stress in Atherosclerosis-Prone
Hypercholesterolemic Mouse

Gilberto L. Pardo-Andreu
a
, Bruno A. Paim
b
, Roger F. Castilho
b
, Jesus A. Velho
b
, Ren Delgado
a
, Anibal E.
Vercesi
b
, Helena C.F. Oliveira
c

a
b
Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas, 13083-887 Campinas, SP,
Brazil.
c
Departamento de Fisiologia e Biofsica, Instituto de Biologia, Universidade Estadual de Campinas,13083-970 Campinas, SP, Brazil.

Atherosclerosis is linked to a number of oxidative events ranging from low-density lipoprotein (LDL) oxidation
to the increased production of intracellular reactive oxygen species (ROS). We have recently demonstrated
that liver mitochondria isolated from the atherosclerosis-prone hypercholesterolemic LDL receptor knockout
(LDLr
-/-
) mice have lower content of NADP(H)-linked substrates than the controls and, as consequence, higher
sensitivity to oxidative stress and mitochondrial membrane permeability transition (MPT). In the present work,
we show that oral supplementation with the antioxidants Mangifera indica L extract (Vimang
) or its main
polyphenol mangiferin shifted the sensitivity of LDLr
-/-
liver mitochondria to MPT to control levels. These in vivo
treatments with Vimang
and mangiferin also significantly reduced ROS generation by both isolated LDLr
-/-

liver mitochondria and spleen lymphocytes. In addition, these antioxidant treatments prevented mitochondrial
NAD(P)H-linked substrates depletion and NADPH spontaneous oxidation. In summary, Vimang
and
mangiferin spared the endogenous reducing equivalents (NADPH) in LDLr
-/-
mice mitochondria correcting their
lower antioxidant capacity and restoring the organelle redox homeostasis. The effective bioavailability of these
compounds makes them suitable antioxidants with potential use in atherosclerosis susceptible conditions.

Key: A27
Key words: Vimang; mangiferin; atherosclerosis; hypercholesterolemia; antioxidant; oxidative
stress; mitochondria


Havana Redox 2009

Antioxidants
Protective Effects of Mangifera indica L extract (Vimang), and Its Major
Component Mangiferin, on Iron-Induced Oxidative Damage to Rat Serum
and Liver

Gilberto L. Pardo-Andreu
a
, Mariela Forrellat Barrios
b
, Carlos Curti
c
, Ivones Hernndez
a
, Nelson Merino
a
, Yeny
Lemus
a
, Ioanna Martnez
a
, Annia Riao
a
, Ren Delgado
a
.

a
Centro de Estudios para las Investigaciones y Evaluaciones Biolgicas. Instituto de Farmacia y Alimentos .Universidad de La
Habana. Ave. 23 # 21425 e/ 214 y 222, La Coronela, La Lisa. CP 13600, Ciudad Habana, Cuba.
b
Instituto de Hematologa e Inmunologa. Apartado Postal 8070, Ciudad de La Habana, CP 10800, Cuba
c
Departamento de Fsica e Qumica, Faculdade de Cincias Farmacuticas de Ribeiro Preto, Universidade de So Paulo, 14040-903
Ribeiro Preto, SP, Brazil

In vivo preventive effects of a Mangifera indica L extract (Vimang) or its major component mangiferin on iron
overload injury have been studied in rats given respectively, 50, 100, 250 mgkg
-1
body weight of Vimang, or
40 mg kg
-1
body weight of mangiferin, for 7 days prior to, and for 7 days following the administration of toxic
amounts of iron-dextran. Both Vimang or mangiferin treatment prevented iron overload in serum as well as
liver oxidative stress, decreased serum and liver lipid peroxidation, serum GPx activity, and increased serum
and liver GSH, serum SOD and the animals overall antioxidant condition. Serum iron concentration was
decreased although at higher doses, Vimang tended to increase it; percent tranferrin saturation, liver
weight/body mass ratios, liver iron content were decreased. Treatment increased serum iron-binding capacity
and decreased serum levels of aspartate-amine transferase (ASAT) and alanine-amine transferase (ALAT), as
well as the number of abnormal Kupffer cells in iron-loaded livers. It is suggested that besides acting as
antioxidants, Vimang extract or its mangiferin component decrease liver iron by increasing its excretion.
Complementing earlier in vitro results from our group, it appears possible to support the hypothesis that
Vimang and mangiferin present therapeutically useful effects in iron overload related diseases.

Key: A28
Key words: Mangifera indica L.; Vimang; mangiferin; iron; reactive oxygen species; antioxidants.


Havana Redox 2009

Antioxidants
Antioxidant Capacity Determination in Mandarin, Maracuya and Grape
Juices

Guel Silva Gabriela, Domnguez Gutirrez Margarita Concepcin, Martnez Morales Lisbeth, Herrera Martnez
Luis Ral, Laura Mndez, Ovando Cindy, Bandala Rodrguez, Eduardo Gmez Conde, Hernndez Castillo
Ma. Isabel

Benemrita Universidad Autnoma de Puebla, Puebla, Mxico

Biological oxidation is induced by active oxygen and mediated by free radicals. Antioxidant capacity of fruits
and vegetables comes from fitonutrients like vitamin C, B, -carotene and polyphenols which have a protective
effect in prevention of degenerative processes in carcinogenic and cardiovascular diseases. Objective of this
work was to determine antioxidant capacity in mandarin, maracuya and grape juices. A cross-sectional
experimental study was performed; studied juices were: mandarin, grape and maracuya. Juices were
obtained, packaged and cooled in amber color glass bottles, during 48 hours before performing
determinations. To determined secuestrant capacity the reagent 2,2-difenil-1-picrilhidracilo (DPPH) was used,
first OD reading (OD1) was at 30 seconds being this value taken as control, second OD reading (DO2) was
done after 30 min. Results shown antioxidant capacity of mandarin juice (84.8%) >> maracuy (78.3%) >
grape (46.72%). It is concluded that mandarin as well as maracuya are rich in vitamins A, C and Flavonoids
which capture reactive oxygen species. For such reason it is important to consider these fruit juices in a
regular diet in order to protect human organism from the action of the reactive oxygen species which cause
diseases and aging.

Key: A29
Key words: Antioxidant, juice, mandarin, maracuya, grape


Havana Redox 2009

Antioxidants
Effect of Aqueous Extract of Yellow Maca in the Levels of Glutathione in the
Brain After an Intense Exercise in Newly Weaned Male Rat

S Surez
a
,
,
R Or
a
L Rojas
b
, R Valdivieso
a
, I Arnao
a
, J Trabucco
a
.

a
Centro de Investigacin de Bioqumica y Nutricin Facultad de Medicina
b
Facultad de Farmacia y Bioqumica; UNMSM. Av. Grau
cuadra 7 s/n, Lima 1, Per.

The Lepidium meyenii Walp (maca) grows at high altitude in the Peruvian Andes, is traditionally used for its
nutritional value and its energizing properties. Objective: To evaluate the glutathione levels in the brain by
administation of the aqueous extract of yellow maca after an intense exercise in newly weaned male rats.
Material and Methods: There were four groups of newly weaned male rats: (I) control, (II) 0.4 mg maca / g
weight (III) 0.8 mg maca / g weight and (IV) 1.2 mg maca / g weight. The aqueous extract was administered by
orogastric canule. It is performed a control of the weights. It was applied the forced swimming test after 30
days of treatment. The animals were sacrificed by decapitation and were prepared brain homogenized to 10%
for the determination of the activity of glucose-6-phosphate dehydrogenase and levels of total glutathione
(GSH). Results: There was no significant difference in weight gain. The resistance test of forced swimming
showed significant difference; were sequentially as follows: (I) 7.25 min; (II) 11.14 min (III) 10.86 min (IV) 13.20
min. Both, the enzyme activity expressed as U / g brain tissue and the levels of total GSH expressed as umol/g
tissue showed no significant differences. Conclusion: The administration of aqueous extract of yellow maca
increases the physical resistance in newly weaned male rats, it seems to be for different mechanisms of the
GSH.

Key: A30
Key words: yellow maca, Lepidium meyenii, GSH, physical resistance


Havana Redox 2009

Antioxidants
Antioxidant Capacity of the Aqueous Extract of Peperomia choroniana
D.CD. (ipintaki) by Means of DPPH and Reception of Hydroxyl Radical.

Olinda Mayhuasca
a
, Silvia Surez
b
, Pablo Bonilla
a

a Institute of Pharmaceutical Sciences and Natural Resources "Juan de Dios Guevara. Faculty of Pharmacy and Biochemistry. b
Research Center of Biochemistry and Nutrition "Alberto Guzman Barron" Faculty of Medicine. Universidad Nacional Mayor de San
Marcos Jr. Puno 1002, Lima 1, Peru , Apartado Postal 4559 Lima 1, Peru ; Telefs: (51-1) 328-4737 annex 30 ; Fax: (51-1) 328-1560

The Peperomia choroniana D.CD. (ipintaki, corazoncito), is a silvester plant, grows in the central region of
peruvian jungle. Traditionally is used to slow the skin aging. Objective: to characterize the chemical
components of the aqueous extract of Peperomia choroniana D.CD. and evaluate its antioxidant capacity.
Material and Methods: Prepared the phytochemical screening l from aqueous extract of rood and leaves of
Peperomia choroniana; the phenolic compounds were characterized by chromatographic and spectroscopic
techniques; was analyze the free radical scavenging capacity of the DPPH radical free and the test of the
hidroxyl free radical sequester by inhibition of the oxidation of the deoxyrribose using Fe
2+
. Results: the
phytochemical screening from aqueous extract of Peperomia choroniana C.DC. showed the presence of
phenolic compounds, quinones and in minor quantity of alkaloids, terpenoids and steroids; of the
chromatogram in thin layer in preparative scale were isolated 6 flavonoids. The watery sample has
demonstrated free radical scavenging capacity of the DPPH free radical, with IC50 of 56.90 ug / mL. Trolox
was used as reference. Also demonstrated an ability to free radical scavenge of the hydroxyl radical of 29.9%
with 150ug/mL of sample. The manitol was used as a reference. Conclusion: The aqueous extract of the air
part of Peperomia choroniana D.CD. contains compounds phenolic as metabolitos secondary predominant,
and have antioxidant activity.

Key: A31
Key words: Peperomia choroniana, antioxidant, DPPH


Havana Redox 2009

Antioxidants

Ferric Reduction Antioxidant Power evaluation on Murraya paniculata L
(Jack) tincture.

C .Casado, Y. Gutirrez, and R. Lpez.

Pharmacy and Food Institute. University of Havana. 23 Ave # 21425 b/ 214 and 222. La Coronela. La Lisa. Havana City. Cuba.
celia@uh.cu; celiamcm@ifal.uh.cu.

Medicinal plants have become a source of new and more efficient antioxidants, to which pharmacology can
appeal to neutralize free radicals produced by cells. Moreover antioxidants have been related to several
illnesses, it has been accepted that those plants that have been traditionally used by their anti-inflammatory
effects, could have a therapeutic action due, at least partly, to the presence of compounds able to react with
free radicals involved in this kind of ailments.
The tincture obtained from Murraya paniculata fresh leaves, used as analgesic frictions, which possesses,
according to previous results, anti-inflammatory properties, presents, taking in to account the bibliographical
reports consulted, an important group of secondary compounds that could be associated to the antioxidant
effect of this phytopharmaceutical. In this sense the drug and the formulation were subjected to a
phytochemical screening that allows us to suppose the presence of alkaloids, terpenoids, coumarins,
flavonoids, among others. Keeping in mind these elements, antioxidant potential of the species were
determined by means of FRAP (Ferric Reduction Antioxidant Power) methodology. The obtained results
showed us a reduction power above 200 , so we can conclude this phytopharmaceutical posses antioxidant
property under the rehearsed experimental conditions.

Key: A32
Key words: FRAP, Murraya paniculata

Havana Redox 2009

Antioxidants
Biological activity and chemical composition of Morinda citrifolia liin and
Annona muricata extracts

Rubio Pino J.L, Iloki Assanga S.B, Lewis Lujn L. M, Meza Cueto C.Y, Gil-Salido A.A, Acosta Silva A.L, Rivera
Castaeda E.G.

Laboratorio de Investigaciones en Bioactivos y Alimentos Funcionales (LIBAF) de la Empresa Rubio Pharma y Asociados S.A. de C.V.
Blvd. Garca Morales km 6.5. Col. El Llano. C.P. 83210, Hermosillo, Sonora, Mxico

The main objective of this work is the chemical and biological characterization of fruit extracts and leaves of
Morinda citrifolia liin and extracts of leaves of Annona muricata. Inside the biological activity the cytotoxic
activity was evaluated in cellular lines of mouse (M12 AK, L929) and human (A549, HeLa) by means of the
spectrophotometric method of reduction of the tetrazolium salt (MTT). The antioxidant activity was quantified
by 2, 2-dipheniylpicrylhydrazyl radical scavenging method (DPPH) and by ferric reducing antioxidant power
(FRAP). As for the chemical composition of the extracts, it was determined the total phenolic content by Folin-
Ciocalteau method and total flavonoid content by Dowd method. The preliminary phytochemical screening was
carried out according to the method established by Rondina and Coussio. The extract of the fruit of Morinda
citrifolia liin and the extracts of leaves of Annona muricata showed in vitro cytotoxic activity against lung and
cervix system cancer cultured cells lines. The extracts of leaves of Annona muricata present an IC
50
50 g/mL,
while the IC50 of the aqueous extract of the fruit of Morinda citrifolia liin was 120 g/mL.
The extracts of leaves of Annona muricata and the aqueous extract of Morinda citrifolia liin were found to
exhibit highest antioxidant activity. High total phenolic content was determined in the extracts of leaves of
Annona muricata. The flavonoid content belonging to the flavanononas families and flavonoles were in more
quantity in the extracts of leaves of Morinda citrifolia liin continued by the extracts of leaves of Annona
muricata.

Key: A33
Key words: FRAP, Morinda citrifolia, Annona muricata

Havana Redox 2009

Redox Signaling Mechanisms
Biomarkers and Diagnostic Methods
Pharmacological or Dietetic Interventions
Other

Index

Redox Signaling Mechanisms................................................................................................Error! Marcador no definido.
Expression, Functionality and Relevance of Sodium-Vitamin C Cotransporter 2 (SVCT2) in Breast Cancer. ............ 100
DHA Reductases and Intracellular Vitamin C Accumulation and Recycling.....................Error! Marcador no definido.
Application of NFkappaB Reporter Assays for Detection of Oxidative Challenge in Human Cancer Cell Lines. ... Error!
Role of Cystein Residues in Modulating Ascorbic Acid Transport by SVCT1...................Error! Marcador no definido.
Regulation of Heat Shock-Induced Apoptosis by Sensitive to Apoptosis Gene Protein...Error! Marcador no definido.
Effect of Glutathione Depletion by Buthionine Sulfoximine on Oxidative Metabolism and Cell Death in Rat Brain.Error!
Uncoupling and Oxidative Stress in Liver Mitochondria Isolated From Rats With Acute Iron Overload Error! Marcador
no definido.
Role of hydrogen peroxide in vitiligo.................................................................................Error! Marcador no definido.
Ozone oxidative preconditioning and postconditioning in acute nephrotoxicity induced by cisplatin in rats........... Error!
Biomarkers and Diagnostic Methods .....................................................................................Error! Marcador no definido.
Evaluation of Nitric Oxide Produced by Immune Cells for Determining Biological Activity in Vaccines Containing
Whole-Cell Pertussis.........................................................................................................Error! Marcador no definido.
Easy High Performance FRAP Utilizing Photocolorimetric Microplate (Format 96 Wells). ...............Error! Marcador no
definido.
Optimisation of an EPR Method for Detection of Intracellular Superoxide Radicals in Stimulated Macrophages.. Error!
Comet Assay: Application to Evaluate the DNA Repair Capacity in Individuals with Xeroderma Pigmentosum
Presumptive Diagnosis. ....................................................................................................Error! Marcador no definido.
Pharmacological or Dietetic Interventions..............................................................................Error! Marcador no definido.
In Vitro Effect of Trofin, an Hydrolyzed of Proteins and Heme Iron Mixture on Ferrous Iron Status... Error! Marcador
no definido.
Estimation of food ingestion and total antioxidant capacity of the offered diet in two elderly homes in the capital Error!
Other ......................................................................................................................................Error! Marcador no definido.
Chenopodium ambrosioides: Antiparasitic Drug that Acts on Mitochondrial Respiratory Chain.......Error! Marcador no
definido.
Oxidative Stress Associated Parameters in Cytotoxicity Induced by 1-O-alkylglycerols in N2a Cell Line.............. Error!


Havana Redox 2009

Expression, Functionality and Relevance of Sodium-Vitamin C
Cotransporter 2 (SVCT2) in Breast Cancer.

Muoz-Montesino C, Sotomayor K, Pea E Sobarzo A, Villagrn M, Soliz C, Barra V, Oate S, Rivas C I, Vera
JC.

Departamento de Fisiopatologa, Facultad de Ciencias Biolgicas, Universidad de Concepcin. Chile

Two functional isoforms of sodium-vitamin C co-transporters (SVCT) have been identified until now: SVCT1
and SVCT2, which mediates the uptake of ascorbic acid (AA) from the extracellular medium. In breast cancer
tissues, there is no expression of SVCT1, and in contrast SVCT2 is widely expressed when compared to
normal breast tissues. The same phenomenon is observed in breast cancer cell lines. Although there is a high
expression of SVCT2 in breast cancer tissues and cell lines, uptake assays in this model showed that these
cells are not capable to transport AA from the extracellular medium, and vitamin C uptake occurs only under its
oxidized form, through GLUT transporters. To understand the relevance of SVCT2 expression in breast
cancer, we performed colocalization analysis to identify the subcellular distribution of this transporter. We
found that SVCT2 is a mitochondrial protein in all breast cancer cell lines analyzed: ZR-75, MCF-7, MDA-MB-
231 and MDA-MB-468, and that this same distribution is seen in breast cancer tissues. At the same time, we
performed western blot analysis for SVCT2 and different organelle markers for subcellular fractions,
concluding that SVCT2 is widely associated with the mitochondrial fraction. To further investigate the role of
SVCT2, this fraction was used to perform AA uptake assays. We found that mitochondria from breast cancer
cell lines are able to transport ascorbic acid, in both extracellular and intracellular conditions. We postulate that
SVCT2 is a functional transporter in breast cancer that mediates the uptake of ascorbic acid in the
mitochondria, and that it might be relevant for breast cancer cell survival under oxidative stress conditions.
Financed by Anillo de Ciencia y Tecnologia Grant ACT28.

Key: RSM1
Key words: sodium-vitamin C co-transporters, breast cancer, mitochondria


Havana Redox 2009

DHA Reductases and Intracellular Vitamin C Accumulation and Recycling

Sotomayor K, Muoz-Montesino C, Ziga F, Sobarzo A, Pea E, Villagrn M, C Soliz-Soto Toro C, Barra V,
Oate S, Rivas C I, Vera JC.

Departamento de Fisiopatologa, Facultad de Ciencias Biolgicas, Universidad de Concepcin. Chile

Vitamin C is an important hydrosoluble antioxidant and a cofactor for several physiologically relevant enzymes.
The human body is incapable to synthesize vitamin C, therefore body tissue levels depend on both the daily
dietary intake and the cellular capacity for vitamin C uptake. Cellular vitamin C transport is mediated by two
types of membrane transporters: the isoforms of the family of sodium-vitamin C cotransporters SVCT1 and
SVCT2, which transport the reduced form of vitamin C or ascorbic acid (AA), and several isoforms that belong
to the family of hexose transporters named GLUTs, which transport the oxidized form of vitamin C or
deydroascorbic acid (DHA). The intracellular management of vitamin C depends greatly on the expression of
DHA reductases, which are enzymes that catalyze the reduction of dehydroascorbic acid back to asocrobic
acid, therefore avoiding the hydrolysis of DHA and the consequent loss of vitamin C. These enzymes are
classified into two major groups: NADPH dependent DHA reductases and glutathione dependent DHA
reductases. In our laboratory, we have cloned and expressed in HEK 293 cells several DHA reductases both
NADPH and glutathione dependent: glutaredoxin 1 (Grx1), glutaredoxin 2 (Grx2), thioredoxin reductase 1
(TrxR1), thioredoxin reductase 2, glutathione-S-transferase 1 (GSTO1) and 2 (GSTO2). We have determined
the localization of these enzymes, the capacity of these enzymes to accumulate vitamin C and the participation
of these enzymes and high vitamin C concentrations in the survival of cells against oxidative stress events. We
conclude that different levels of expression of these enzymes can be correlated with a different cellular
capacity to manage vitamin C and therefore can be implicated with a different cellular response towards
oxidative stress.

Key: RSM2
Key words: deydroascorbic acid reductases, Vitamin C, oxidative stress


Havana Redox 2009

Application of NFkappaB Reporter Assays for Detection of Oxidative
Challenge in Human Cancer Cell Lines.

L Pulaski,
a,b
I Sachrajda
b
, D Jaros
b
, G Bartosz
a

a
Department of Molecular Biophysics, University of Lodz, Banacha 12/16, 90-237 Lodz, Poland.
b
Laboratory of Transcriptional
Regulation, Institute of Medical Biology PAS, Lodowa 106, 93-232 Lodz, Poland.

Artificial reporter constructs are excellent tools for studying the activation of NFkappaB by reactive
oxidant signals. We have constructed novel reporter plasmids based on two detection modes:
luminometric (with luciferase as the reporter gene) and microscopic (with advanced fluorescent
proteins as reporter genes). The expression of reporter proteins is transcriptionally regulated by
artificial promoters consisting of HSV TK or adenovirus E1b TATA box (minimal promoter) and six
concatenated NFkappaB consensus response elements upstream of it. This ensures relatively low
basal level of reporter gene expression and robust enhancement of reporter presence upon
NFkappaB activation, which can be detected already several hours after the stimulus. The reporter
constructs were transfected into human cancer cell lines Sk-Ov-3 (ovarian carcinoma), A549 (lung
adenocarcinoma), MCF-7 (breast carcinoma) and HeLa (cervical carcinoma), known for their ability to
upregulate NFkappaB activity significantly and for their use as cellular models of oxidative stress
response. Activation of the NFkappaB pathway by exogenous oxidative challenge is a controversial
issue, with some studies having shown clear-cut stimulation of this stress response pathway by
reactive oxidants, while other results pointed rather to the inhibitory influence of redox imbalance on
NFkappaB activity. Our studies, aimed at thorough validation of the NFkappaB signal in varying
cellular background, reactive oxidant concentration and time scale, led to the overall conclusion that
hydrogen peroxide (a model natural reactive oxidant) in most cell lines has a triphasic response curve
of NFkappaB signalling, with two distinct (although not very prominent) peaks of activatory
concentrations and a concentration range where there was conspicuous inhibition of signal even
though cell viability was not yet compromised. Most interestingly, the specific range of activatory
concentrations was cell type-specific and correlated well with some aspects of cellular antioxidant
defence capability (oxidative stress resistance, andioxidant enzyme expression), while it was
unrelated to the basal activity of the NFkappaB pathway itself.

Key: RSM3
Key words: NFkappaB signalling, hydrogen peroxide, cancer

Havana Redox 2009

Role of Cystein Residues in Modulating Ascorbic Acid Transport by SVCT1

Haensgen H, Zuiga F, Rivas CI, Vera JC

Facultad de Ciencias Biolgicas, Departamento de Fisiopatologa, Universidad de Concepcin, Concepcin, Chile.

The SVCT1 transporter mediates the transport of ascorbic acid (AA) by a sodium-ascorbate cotransport
mechanism. SVCT1 has a 100 M km for AA and is cooperativly activated by the presence of sodium ions
(Na
+
) and modulated by the presence of divalent cations (Ca
2+
, Mg
2+
). Nevertheless, although the functional
characterization of this transporter is very advanced there is no available structural information for this
transporter that can allow the establishment of a structure-function relationship for SVCT1. The literature
describes a very important structural and functional role for cysteine residues in membrane proteins,
suggesting the importance of these residues in SVCT1. We analyzed the primary structure of SVCT1 and
found 15 cysteine residues along its sequence that were localized in transmembrane domains, exofacial and
endofacial loops. We evaluated the importance of these residues by three methods. We expressed the
SVCT1-GFP chimera in HEK-293 cells and analyzed the effect of both cysteine reduction and cysteine
chemical modification using correspondingly DTT and PCMB. Third, we analyzed the importance of each
cysteine residue with site directed mutagenesis changing each of these residues to serine and analyzing the
effect of these mutants on the capacity of AA transport and the subcellular localization of SVCT1. A confocal
microscopy analysis revealed that all mutants displayed the expected cell membrane localization.
Nevertheless, three mutants displayed an impaired capacity to transport AA, transporting only 50% as much
as SVCT1.Even though DTT treatment without prior incubation didnt affect the function of SVCT1, a 30-min
preincubation with this reducing agent, as well as PCMB treatment, induced an important loss of AA transport
activity of SVCT1 in a dose-dependent manner. All this taken together suggests an important role for cysteine
residues in SVCT1s function.

Proyecto Anillo de Ciencia y Tecnologa ACT-28, Red 26 y beca CONICYT de doctorado

Key: RSM4
Key words: cysteine reidues, SVCT1 transporter, ascorbic acid


Havana Redox 2009

Regulation of Heat Shock-Induced Apoptosis by Sensitive to Apoptosis
Gene Protein

Jeen-Woo Park, Sun Joo Lee, Eun Sun Yang, Sung Youl Kim, Seoung Woo Shin

School of Life Sciences and Biotechnology, Kyungpook National University, Taegu 702-701, South Korea

Heat shock may increase oxidative stress due to increased production of reactive oxygen species and/or the
promotion of cellular oxidation events. Sensitive to apoptosis gene (SAG) protein, a novel zinc RING finger
protein that protects mammalian cells from apoptosis by redox reagents, is a metal chelator and a potential
reactive oxygen species scavenger, but its antioxidant properties have not been completely defined. In this
report, we demonstrate that modulation of SAG expression in U937 cells regulates heat shock-induced
apoptosis. When we examined the protective role of SAG against heat shock-induced apoptosis with U937
cells transfected with the cDNA for SAG, a clear inverse relationship was observed between the amount of
SAG expressed in target cells and their susceptibility to apoptosis. We also observed the significant decrease
in the endogenous production of reactive oxygen species and oxidative DNA damage in SAG-overexpressed
cells compared to control cells upon exposure to heat shock. In addition, transfection of PC3 cells with SAG
small interfering RNA markedly decreased the expression of SAG, enhancing the susceptibility of heat shock-
induced apoptosis. Taken together, these results indicate that SAG may play an important role in regulating
the apoptosis induced by heat shock presumably through maintaining the cellular redox status.

Key: RSM5
Key words: heat shock, apoptosis, redox status


Havana Redox 2009

Effect of Glutathione Depletion by Buthionine Sulfoximine on Oxidative
Metabolism and Cell Death in Rat Brain.

ME Gonzlez Fraguela, L Blanco Lezcano, L Lorigados Pedre, R Garca Miniet

Internacional Center of Neurological Restoration. Ave 25 No 15805. Playa. Ciudad de la Habana. Cuba

Our purpose was to test the effects of depleting tissue glutathione (GSH) by buthionine sulfoximine (BSO,
10mM, intracerebroventricular (icv)) on brain oxidative metabolism indicators and DNA fragmentation in rat
brain. The experimental groups were: icv BSOCtreated at 24h and 48h, icv saline-treated at 24h and 48h and
untreated groups. 24h after BSO treatment, GSH levels in hippocampus dropped down to 55% as compared to
untreated animals, 48h later the hippocampal GSH values showed a slight increase, in contrast, had been
significantly restored in striatum and frontal cortex. The administration of BSO induced significant changes in
antioxidant enzymatic activities like superoxide dismutase, catalase and glutathione peroxidase and modified
the malonilaldehyde content and tumor necrosis factor levels as direct indicators of oxidative cellular
damage. This fact was especially noticeable in hippocampus. DNA fragmentation study for
inmunohistochemical techniques detected TUNEL + cells in the hippocampal area in animal treated with BSO
at 6h, 12h and 24h and this contrast whit a very few cases of TUNEL + cells in the saline groups. Our results
support that glutathione status is a key piece acting in the regulation of brain function.

Key: RSM6
Key words: glutathione, buthionine sulfoximine, brain


Havana Redox 2009

Uncoupling and Oxidative Stress in Liver Mitochondria Isolated From Rats
With Acute Iron Overload

Gilberto L Pardo Andreu
a
, Natalia M Inada
b
, Anibal E Vercesi
b
, Carlos Curti
c
a
b
Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas 13083-970 Campinas, SP,
Brazil
c
Departamento de Fsica e Qumica, Faculdade de Cincias Farmacuticas de Ribeiro Preto, Universidade de So Paulo, 14040-903
Ribeiro Preto, SP, Brazil

One hypothesis for the etiology of cell damage arising from iron overload is that its excess selectively affects
mitochondria. Here we tested the effects of acute iron overload on liver mitochondria isolated from rats
subjected to a single dose of i.p. 500 mg/kg irondextran. The treatment increased the levels of iron in
mitochondria (from 21 4 to 130 7 nmol/mg protein) and caused both lipid peroxidation and glutathione
oxidation. The mitochondria of irontreated rats showed lower respiratory control ratio in association with
higher resting respiration. The mitochondrial uncoupling elicited by irontreatment did not affect the
phosphorylation efficiency or the ATP levels, suggesting that uncoupling is a mitochondrial protective
mechanism against acute iron overload. Therefore, the reactive oxygen species (ROS)/H
+
leak couple
functioning as a mitochondrial redox homeostatic mechanism could play a protective role in the acutely iron
loaded mitochondria.

Key: RSM7
Key words: Iron overload; Mitochondria; Oxidative stress; Reactive oxygen species (ROS);
Uncoupling


Havana Redox 2009


Role of hydrogen peroxide in vitiligo

Nubia Fernndez Hernndez
a
, Janeth E. Rojas Urdaneta
b
a

a
Centro de Estudios para las Investigaciones y Evaluaciones Biolgicas. Food Science and Pharmacy College, University of Havana,
Cuba. UH.
b
Jerico c.a. Venezuela.

Vitiligo is a pigmentation disorder with an incidence in 1 and 3 % of the worldwide population. It is studied for
years but the exact causes are not already knows. Vitiligo is characterized for the lack of skin pigmentation
and the presence of not functional melanocites in epidermis.
Different researchers groups try to explain the pathological origin of this disease. However, consent has not
reached. Independent groups defend his theory with solid arguments but without the integration of all the
knowledge already reached about this disease.
This work integrates some of the main findings to a common event: The demonstrated presence of
hydrogen peroxide (H
2
O
2
) in high concentrations and the decrease of catalase in epidermal and serum
in patients with vitiligo.
The normal basal levels of H2O2 in epidermis, measured in keratinocites, overcome the concentration in
melanocites. Disruptions in oxidative stress environment give raise the increment in the concentration of H2O2
in keratinocites and melanocites. Keratinocites act as source of reactive oxygen species (ROS) and transfer to
the melanocites, even by passive diffusion, great quantities of H
2
O
2
.
Experimental data confirm that as a results of the presence of high concentration of H
2
O
2
in aftereffects
epidermis in vitiligo patients:
The levels of butyrylcolinesterase (BchE) and acetylcolyneterase (AchE) decrease;
The dihydropteridine reductase (DHPR) activity, enzyme involved in the cycle of tetrahydrobiopterin
(6HB4), significantly decrease;
The stress's proteins are up-regulated;
The concentration of peptides derived from proopiomelanocortins (POMC) is affected.
All these events, after a triggered situation on a genetically predisposed individual, promote changes in the
skin pigmentation and lead to the vitiligo, as a final resort.

Key: RSM8
Key words: Vitiligo, hydrogen peroxide

Havana Redox 2009

Ozone oxidative preconditioning and postconditioning in acute
nephrotoxicity induced by cisplatin in rats

Silvia Menndez
a
,

Ricardo Gonzlez,
b
Zullyt Zamora,
b
Aluet Borrego,
b
Frank Hernndez
b
and Teresa Montero
c

a
Ozone International Clinic, Ozone Research Center. PO Box 6414, Havana, Cuba. Phone: 537-2719264. E-mail:
silviamenendez@infomed.sld.cu
b
Department of Biomedicine, Ozone Research Center. Havana, Cuba.
c
Luis Daz Soto Surgical and Clinical Hospital, Havana, Cuba.

Cisplatin (CDDP) is one of the most effective chemotherapeutic agents for the treatment of ovarian, testicular and bladder
carcinomas and cancers of the head and neck and lung. The cellular events in CDDP-induced acute nephrotoxicity,
including decreased protein synthesis, apoptosis induction, membrane lipid peroxidation, mitochondrial dysfunction and
DNA injury, are a consequence of free radical generation and the inability to scavenge such molecules. Acute renal failure
induced by CDDP is accompanied by the reduced renal blood flow associated with increase renal vascular resistance and
histological damage to the proximal tubular cells. Also, a depletion of renal reduced glutathione (GSH), an increase of
lipid peroxidation and a decrease in the activity of the antioxidant defence system, are observed. Ozone oxidative
preconditioning and postconditioning is a prophylactic approach, which favors the antioxidant-prooxidant balance for
preservation of cell redox state by increasing antioxidant endogenous systems. The aim of this study is to analyze the
protective role of ozone oxidative preconditioning and postconditioning against cisplatin-induced nephrotoxicity. For both
preconditioning and postconditioning prodecures, male Sprague-Dawley rats were treated with intra-rectal ozone
applications at a dose of 0.36, 1.1 and 1.8 mg/kg. For the first procedure rats were pretreated with 15 ozone applications
before cisplatin intraperitoneal (ip) injection (6 mg/kg) and for the second procedure rats were treated with 5 ozone
applications after cisplatin ip injection. Three control groups were included: Non treated rats; CDDP-rats submitted to the
challenge with CDDP ip injection (6 mg/kg) and Oxygen-rats treated with oxygen instead of ozone in both procedures. In
each group, 8 rats were included. Serum creatinine was measured, as well as thiobarbituric acid reactive substances,
reduced glutathione peroxidase (GSH) content and the activities of antioxidant enzymes (superoxide dismutase-SOD,
catalase-CAT and glutathione peroxidase-GSH-Px) in renal homogenate. The results demonstrated that both, ozone
oxidative preconditioning and postconditioning, diminished the increase in serum creatinine levels and TBARS content,
the GSH depletion and also reversed the inhibition of SOD, CAT and GSH-Px activities induced by cisplatin in the rat
kidney. These protective effects of ozone were dose dependent, obtaining best results with the dose of 1.1 mg/kg. The
histological study demonstrated that the tubular necrosis in the ozone group of each different procedure was slight and in
lesser extent, with significant diminution in the dequamation of tubular epithelial cells in comparison with the CDDP group.
We can conclude that intrarectal ozone applications reversed the renal pro-oxidant unbalance induced by cisplatin
treatment by the way of stimulation to some constituents of antioxidant system in the kidney. Thereby, it decreases the
renal damage as it was demonstrated in the histological study.

Key: RSM9
Key words: Ozone, postconditioning, preconditioning, cisplatin

Havana Redox 2009


Evaluation of Nitric Oxide Produced by Immune Cells for Determining
Biological Activity in Vaccines Containing Whole-Cell Pertussis

Chovel ML
a
, Figueroa JM, Abreu I and Garca M
b

a
Finlay Institute, Avenida 27, No. 19805, La Lisa, CP 11600, Habana, Cuba.
b
Centro para el Control Estatal de la Calidad de los
Medicamentos (CECMED), Calle 200, No. 1706, Entre 17 y 19, Siboney, Playa, CP 11600, Habana, Cuba.

It has been increasingly accepted that Nitric oxide (NO) is an important regulator of the immune system. In
fact, inflammatory cells, including monocytes and resident macrophages produce NO following incubation with
bacteria, bacterial lipopolysaccharides and cytokines. Production of NO from activated murine macrophages
has been implicated as an antimicrobial effector mechanism against several pathogens. The present study
aims to evaluate the suitability of a cell culture test based on the generation of NO by macrophages and
spleen cells from mice for determining biological activity of vaccines containing whole-cell pertussis (wP)
component. For that, ten mice per group were immunised with wP vaccine dilutions and macrophages and
spleen cells were collected after immunisation. Cells were pooled and resuspended in supplemented RPMI
1640 medium. They were cultured in presence of heat-killed (HKC) Bordetella pertussis 18323, a sonic extract
of the same strain and LPS. Nitrite determinations were made by using Griess reagent. IFN concentrations
were measured by ELISA. It was also evaluated if there was a correlation regarding the in vivo challenge test.
The HKC, the sonicate supernatant and LPS induced strong NO synthesis by macrophages and spleen cells
of vaccinated mice. The production of IFN- correlated well with the formation of nitrite. A good correlation was
also obtained against the challenge test in mice. It is not known if NO has a direct bactericidal effect on
Bordetella pertussis, but NO production may serve as a marker of macrophage activation, an important host
defence against this bacteria. The correlation versus the challenge test enables the cell culture test as a
potential alternative to evaluate biological activity of vaccines containing wP, taking into account the limitations
of the in vivo test.

Key: BDM1
Key words: NO, vaccine, Bordetella pertussis

Havana Redox 2009


Havana Redox 2009

Easy High Performance FRAP Utilizing Photocolorimetric Microplate
(Format 96 Wells).

Mercader OA
a
, Rodriguez JL
b
, Arocha C
a

a
Instituto de Nutricin e Higiene de los Alimentos, Infanta 1158, Centro Habana, Cuba.
b
Instituto de Investigaciones de la Industria
alimenticia, Guatao, Lisa, Cuba.

In order to develop an analytical platform of high efficiency for determination of Total Antioxidant Capacity in
foods and plasma for FRAP method, came true the calibration of method described by Benzie and Strain
(1996), utilizing Fe
2+
solutions in the interval of concentrations of 200 to 1000 M and microplate reader
(TecnoSuma, Cuba) in absorbance mode, with 540 nm. Result suggested that the method semiautomatic
utilized have coefficients of regression in the order of 0.99 to check against the linearity of reply with calibration
curve of Fe
2+
and different solution of Galic acid and black tea, that indicates a good analytical performance for
the determinations of technique developed in the INHA.

Key: BDM2
Key words: FRAP, microplate, analytical assay


Havana Redox 2009

Optimisation of an EPR Method for Detection of Intracellular Superoxide
Radicals in Stimulated Macrophages

M Deschacht
a
, T Horemansa, K Tot
a
, L Maesa,
b
P Cos
a

a
Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp,
Belgium,
b
Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerp, Belgium. E.mail: paul.cos@ua.ac.be

Despite the apparently clear relationships between oxidative stress and lung- and cardiovascular
disease, antioxidant therapy so far has not been very successful in either setting. This exemplifies the
need for proper documentation of actual levels of free radicals and of specific and sensitive
biomarkers to establish their potential contribution to pathological processes. However, most
investigators employ indirect approaches and measure markers of oxidative damage to biomolecules
(fingerprint assays). An analytical approach that permits direct detection of free radicals is electron
paramagnetic resonance (EPR) combined with spin trap technique. The aim of our study was to
establish an optimal protocol for the detection of intracellular superoxide radicals by comparing
different spin traps and stimulators at different incubation temperatures and times. PMA, LPS, INF
and TNF were used as stimulators for U937 (monocytes), J774, RAW 264 and primary mouse
macrophages. Three cyclic amine spin traps were compared, with CM-H for intracellular and CP-H
and PP-H for extracellular radical measurements. Optimisation of this EPR spin trap technique
resulted in a direct, sensitive and specific method for the detection of intra- and extracellular
superoxide radicals in different types of stimulated macrophages. The use of PMA as a stimulator (5
min incubation) and CM-H as a spin trap (60 min incubation) gives the highest response in EPR
signals for intracellular superoxide radicals. This optimized method is now being applied to a
Leishmania infection model to investigate the role of oxidative stress in the survival of amastigotes in
the macrophage and in the mechanism-of-action of several antileishmania drugs.

Key: BDM3
Key words: electron paramagnetic resonance, superoxide radical, macrophages


Havana Redox 2009

Comet Assay: Application to Evaluate the DNA Repair Capacity in
Individuals with Xeroderma Pigmentosum Presumptive Diagnosis.

Pupo JB, Gutierrez R, Pandolfi A, Cutara E, Acosta T, Roblejo H, Lantigua A, Pereira N

*National Center of Medical Genetics. Higher Institute of Medical Sciences of Havana. Address: 146 No. 3102. Playa, PC 10600,
Havana City, Cuba. Phone: (53) (7) (208) 9991 - 9999 ext.1093.

Introduction. Xeroderma Pigmentosum (XP) is an uncommon autosomal recessive disease; it is caused by
genes mutations implied in DNA nucleotide excision repair (NER). XP is associated to ultraviolet light (UV)
extreme cutaneous photosensitivity and a high risk to develop skin cancer. REN kinetic was evaluated and
compare between normal individuals and presumptive XP patients using alkaline comet assay in isolated
lymphocytes. Materials and Methods. The study universe comprehends 29 patients and 40 controls subjects.
Isolated lymphocytes from both groups were exposed to a single and identical uvC dose and incubated during
0, 45 and 180 min. Results and Conclusions. Healthy individuals showed a two-phase behaviours REN
kinetics: there was a significant increment of DNA migration at 45 min of incubation (first step), followed by a
quick decline at 180 min (second step), reaching similar values to the basal ones. DNA migrations increase is
attributed to the excision activity on UV-damaged DNA. Polymerization and ligation steps subsequently rejoin
the broken ends, determining a reduction of DNA. In presumptive XP patients, there were individuals that
followed a two-phase kinetics. In others patients, the REN kinetics first step was affected, probably due to a
disruption of some of the proteins implied in DNA damages recognition and excision steps; while in some
individuals the second step was not observed. In this group, the deficiency could be located in proteins
involved in the polymerization and binding of the breach. Alkaline comet assay allows following individual REN
two-phase kinetics, being very useful to evaluate the DNA repair capacity of individuals with suspicion of
deficient REN, like in XP patients.

Key: BDM4
Key words: Xeroderma pigmentosum, nucleotide excision repair, uv-damaged, DNA


Havana Redox 2009

In Vitro Effect of Trofin
, an Hydrolyzed of Proteins and Heme Iron Mixture

on Ferrous Iron Status

Y. Garca,
a
Gilberto L Pardo-Andreu,
b
D. Martnez
a
and R. Gonzlez
a

a
National Center of Biological Products (BIOCEN) Beltrn Mainroad, 1 km, Bejucal, Havana. Cuba. Email Yenela@biocen.cu
b

Centro de Estudios para las Investigaciones y Evaluaciones Biolgicas. Instituto de Farmacia y Alimentos .Universidad de La
Habana. Ave. 23 # 21425 e/ 214 y 222, La Coronela, La Lisa. CP 13600, Ciudad Habana, Cuba.

Iron salts as source of ionic iron are broadly used for the prevention and treatment of iron deficiencies, but they
cause adverse reactions in 25% of the patients. Trofin is a natural formulation containing blood caw as heme
iron source and proteins. It has been evaluated in different population groups, with a high effectivity and
absence of adverse reactions. It is well known that there are two different receptors in the duodenum for the
absorption of the ionic and the heme form of iron and that ferrous state is necessary for ionic iron absorption.
Autoxidation of Fe
+2
is affected by chelation, pH, buffers and other heavy metals. In this regard, a formulation
containing the two type of iron at the same time could favor total iron absorption if it is capable to preserve the
ionic iron in its reduced state. The aim of this work was to evaluate the influence of Trofin on oxidation of
Fe
+2
. It were prepared four different combination of Trofin +Fe
+2
in a w/w relation of: 500:1, 2 500: 1, 5000:1
and 10 000:1, each containing 20 M Fe
+2
in 50 mM HEPES buffer, pH=6,5. The remaining Fe
+2
concentration
was determined at 0, 5, 10, 15, 20, 25 and 30 min of Trofin-iron incubation using 1 mM 1,10 Phenantroline
and the spectrophotometric detection at 510 nm. Results show that Trofin dose dependently caused a
diminution of Fe
2+
concentration The amounts of iron diminution after 30 min of incubation were around 10, 20,
40 and 60% at 500:1, 2 500: 1, 5000:1 and 10 000:1 Trofin-Fe w/w ratios, respectively. Further experiments
will be require to elucidate the mechanism of Trofin-ionic iron interaction, but probably some Trofin
constituents stimulated the oxidation iron. These results suggest that the development of new antianemic
formulation containing Trofin+ Fe
+2
should be observed cautiously and should comprise amounts of Trofin
below the proportion 500:1 in relation to iron.

Key: PDI1
Key words: Trofin, iron, autoxidation

Havana Redox 2009


Estimation of food ingestion and total antioxidant capacity of the offered diet
in two elderly homes in La Habana

Bilbao, T., Forbe, T. and Prez, Y.

Institute of Pharmacy and Foods, Havana University

The aim of this research was to estimate the quality of the diet in two elderly homes (I and II) in the capital,
both for the content and quality of the main components of the diet, as well as for the total antioxidant capacity
(TAC) of the diet foods. Food consumption was determined by the visual appreciation method (Gay et al.,
1992) and energy and nutrients ingestion by the CERES computation method, version 1.02 (FAO, 1997). TAC
values were estimated from the individual values of antioxidant capacity in the average diet offered in each
elderly home of the published foods by Halvorsen et al., 2006, and the quantities of the consumed foods by
senior adults. Results showed that the diet of the elderly in both homes neither were balanced, nor enough,
adequate and varied, and was characterized by a marked deficiency of fruit consumption and almost nothing
of vegetables; this explained the failure in attaining the recommendations of most vitamins and minerals. The
average diet TAC showed significant differences (p<0.05) between the two elderly homes, where the higher
value was 4.92 1.36 (elderly home II). According to the Cuban Food Guide, the food groups of major
contribution to this value were Group I: Cereals and viands, and Group III: Fruits, and the values obtained from
coffee consumption.

Key: PDI2
Key words: CERES, diet, total antioxidant capacity


Havana Redox 2009

Other
Chenopodium ambrosioides: Antiparasitic Drug that Acts on Mitochondrial
Respiratory Chain

L. Monzote,
a
L. Gille
b

a
Institute of Tropical Medicine Pedro Kour, Apdo No. 601, Marianao 13, Havana City, Cuba.
b
Molecular Pharmacology and Toxicology
Unit, Department of Biomedical Sciences, University of Veterinary Medicine, Vienna, Austria.

The diseases caused by parasitic infections are responsible for considerable morbidity and mortality. The
progress in the development of vaccines against parasite infections tends to be slow and the epidemiological
control of diseases is unsatisfactory. Currently, the chemotherapy remains an essential component of clinical
management and disease control programs. A number of factors limit the benefit of existing drugs, such as low
efficacy, poor compliance, high toxicity and drug resistance. Accordingly, the demand for new therapies is
continuous. The majority of people in endemic areas of leishmaniasis depend of natural and traditional
medicine to alleviate the symptoms. In previous work, the antiparasitic activity of the essential oil from
Chenopodium ambrosioides was demonstrated against Leishmania amazonensis, Plasmodium falciparum and
Trichomona vaginalis. In this study, the possible mechanism of action of the Chenopodium oil and their main
components (ascaridole, carvacrol and caryophyllene oxide) was elucidated. Evidence for the paramagnetic
one-electron, centering in carbon radical has been showed by Electron Spin Resonance techniques in the
essential oil, due to the ascaridole. The tested products inhibited the electron transport in Complex I and II of
submitochondrial particles isolates from bovine liver (Bovine taurus) as mammalian model or yeast
(Saccharomyces cerevisiae) as surrogate protozoa model. However, the Chenopodium oil showed an
inhibition statistically significant against Complex III or bc1 from yeast compare with mammalian bc1 Complex.
These data are evidences that supports that the antiparasitic mechanism of action of essential oil from C.
ambrosioides can be result of the formation of a free radical that inhibit the electron transport chain of the
mitochondria.

Key: O1
Key words: Chenopodium ambrosioides, mitochondria, antiparasitic


Havana Redox 2009

Other
Oxidative Stress Associated Parameters in Cytotoxicity Induced by 1-O-
alkylglycerols in N2a Cell Line.

Marian Hernndez Colina, Yolanda Valds Rodrguez and Jos Luis Len

Instituto de Farmacia y Alimentos, Universidad de la Habana. Ave 23 No. 21425 e/214 y 222, La Coronela, La Lisa, Ciudad de la
Habana, Cuba

1-O-alkylglycerols has been studied from two decades ago in our country, due to their potential
activity over inflammation, metastasis and angiogenic processes. Obtained by chemical synthesis in
Basic Chemistry Department of Pharmacy and Food Institute, at University of Havana, their
properties have been demonstrated by our workgroup. Those results include cell differentiation,
apoptosis at higher concentrations, and concentration dependent cytotoxicity. The aim of this study
was to evaluate the influence of oxidative stress in cytotoxicity mediated by 1-O-alkylglycerols. Levels
of nitric oxide, reduced glutathione, total organic peroxides and malonildialdehyde were measured in
supernatants of N2a cell cultures, previously treated with cytotoxic and differentiating concentrations
of decyl, dodecyl, hexadecyl and octadecylglycerol. Marked decrease in total nitrites was found for all
assayed concentrations, with no changes in glutathione levels. Total organic peroxides were elevated
at cytotoxic concentrations and also MDA levels. With those results, we can conclude that oxidative
stress influences the cytotoxicity of 1-O-alkylglycerols in N2a cells, but independently of
concentration. It seems that cytotoxic damage is mediated by oxidative membrane damage and
oxidant peroxides generation, but coexist other factors that influence the cytotoxicity of those
compounds in N2a cells.

Acknowledgements: Studies Centre for Biological Research and Evaluations, Pharmacy and Food Institute, University of Havana.

References: 1. Valds Y (1997) Tesis de doctorado, UH. 2. Chandra J et al. (2000) Triggering and modulation of apoptosis by
oxidative stress. Free Radical Biology & Medicine, 29(3/4): 323-33. 3. Sotomayor H et al. (2006) Citotoxicidad de 1-O-decilglicerol y 1-
dodecilglicerol sintticos sobre carcinoma humano de mama MCF-7. Acta Farm. Bonaerense.; 25(3):339-43.

Key: O2
Key words: 1-O-alkylglycerols, oxidative stress, cytoxicity

Havana Redox 2009

Authors Index

A Gutirrez....................93
A Pereira ................44, 83
A. Gil ............................81
A. Hernndez-Mijares ....76
Abreu I........................123
Abreu P ......................103
AC Vieira ......................83
Acosta Silva A.L ..........111
Acosta T....................127
Adriana Filip..................23
Adyari Fallarero.............96
Akel Mallok ...................70
Alain Valdivia Acosta......56
Alberto J. Nez Sells ..27
Alexis Vidal ...........94, 96
Aluet Borrego ..............122
AM Moratelli ..................40
Amanda Melillo..............22
Ana Elsa Batista ............96
Andres Melendez...........22
Andriy Samokhin ...........96
ngela Fraga ................61
Anibal E Vercesi ..105, 120
Anna Cabr...................68
Annia Riao ................106
Antoni Castro ................68
Ariadna Costa ...............96
Arocha C ....................125
Arqumidez L. Daz Batista
................................59
Ballesteros M................53
Bandala Rodrguez ......107
Barra V...............113, 114
Batista AE.....................94
Bilbao, T...................129
Boram Nho....................96
Bruno A. Paim.............105
C .Casado...................110
C Santacruz Vazquez ....79
C Soliz-Soto Toro C.....114
C. Bauls...................... 76
C. Oktenli.......... 37, 38, 39
C.Tatomir...................... 23
Cabrera-Gmez JA.. 58, 66
Cardosa E..................... 72
Cardosa P..................... 72
Caridad Margarita Garca
Pea ........................ 62
Carlos Curti ......... 106, 120
Casanova-Orta M.... 58, 66
Celia A. Alonso Rodrguez
................................ 70
Cervantes-Llanos M. 58, 66
Chacon A.L................... 97
Chovel ML .................. 123
Cindy Bandala Rodrguez
................................ 77
Claudina Zaldvar .... 94, 96
CM Oliva....................... 63
CM Ribeiro.............. 14, 42
Correia JFG.................. 83
Costa A ........................ 94
Costa D...................... 103
Cutara E.................. 127
D Alonso..... 51, 57, 63, 65
D Castro ....................... 90
D Gonzlez................... 90
D Jaros....................... 116
D Kaur .......................... 12
D Lee ........................... 12
D Wilhelm Filho 21, 40, 42,
44, 83, 85
D. Martnez
..................... 128
Danae Prez Santana.... 91
DB Incio...................... 40
Dieter Brmme.............. 96
Domnguez Gutirrez
Margarita Concepcin77,
107
E Moreno.... 51, 57, 63, 65
E Pedroso... 51, 57, 63, 65
EA Ferreira ....... 21, 44, 83
EB Parisotto 14, 40, 42, 44
Eduardo A. Ferreira ....... 85
Eduardo Gmez ............ 77
Eduardo Gmez Conde107
Eduardo Pentn reas... 17
Ela Cspedes................ 68
Eliana F. Gris ................ 85
EM Dalmarco ................ 14
Emilio E. Monteagudo
Jimnez.................... 95
Ernesto Vega ................ 92
Eun Sun Yang............. 118
Evangelina Marrero ....... 92
F Biscaro ...................... 44
F Dal-Pizzol .................. 40
F Yaheln...................... 64
F. Coll-Manchado.......... 88
Fernandes E ............... 103
Fernndez Y ................. 72
Figueroa JM................ 123
Forbe, T.................... 129
FP Possamai ................ 40
Fraga J.A...................... 97
Frank Hernndez......... 122
FSL Catanhede............. 83
G Bartosz ................... 116
Gabriela Chereches....... 23
Garca JC..................... 89
Garca M .................... 123
Gema Prez Davison.... 60,
100
Gilberto L Pardo Andreu25,
105, 106, 120, 128
Gilberto Snchez-Cruz... 73
Gil-Salido A.A.............. 111
Gmez-Chvez D.......... 58
Gonzlez TL ............... 103

Havana Redox 2009

Gregorio Martnez Snchez
.18, 56, 58, 59, 60, 64,
66, 70, 73, 82, 92, 97,
99, 100, 121
Grzegorz Bartosz...........30
Guel Silva .77, 78, 79, 107
Gutierrez R50, 52, 89, 127
Haensgen H................117
Helena C.F. Oliveira.....105
Hernndez Castillo Ma.
Isabel .....................107
Herrera Martnez Luis Ral
........................77, 107
Hetzel de la C. Lourido
Prez................60, 100
Horta D...................97, 99
I Arnao..................80, 108
I Garca ........................93
I Gonzlez-Blanco .........88
I Sachrajda..................116
Iloki Assanga S.B ........111
Ioanna Martnez ..........106
Iolanda Lazaro ..............68
Ion-Dan Postescu ..........23
Irene Alonso Camaraza..54
Ivones Hernndez........106
J L Rodrguez................90
J Trabucco..................108
J. Borroto....................104
J. Dranguet-Vaillant .......88
J. Londoo....................81
J. Quiones.................104
J. R. Herance ................76
J. V. Esplugues .............76
Janet Snchez ..............92
Janeth E. Rojas Urdaneta
........................82, 121
Janetsy Borroto .............87
Jaya Dasgupta ..............22
Jeen-Woo Park............118
Jesus A. Velho ............105
JK Andersen .................12
JL Rodrguez...............101
Joe A. Vinson................ 11
Jos Luis Len.......... 132
Josefa Girona................ 68
Josep Coll..................... 87
Jussara M. Rebello........ 85
K Tot .................. 10, 126
KB Felipe.... 21, 44, 83, 85
KS Moschetta................ 44
L Blanco Lezcano........ 119
L Ledesma.................... 93
L Maes ....................... 126
L Pulaski..................... 116
L Rojas....................... 108
L. Gille...................... 130
L. Maes ........................ 10
L. Monzote ............... 130
L.Rojas......................... 80
Lantigua A ...... 50, 52, 127
Laura Mndez ....... 77, 107
Lewis Lujn L. M ......... 111
Livan Delgado Roche..... 59
Lluis Masana................. 68
Lopategui-Cabezas I...... 66
Lpez Saura P....... 58, 66
Lorigados Pedre.......... 119
Luis Velzquez-Prez .... 73
M Deschacht
..................... 126
M Kviecinsk................... 44
M Prez........................ 94
M. Deschacht ................ 10
M. Hernndez ............. 104
M. Rocha...................... 76
M.H. Terekeci.... 37, 38, 39
M.R Kviecinski............... 21
Maite Elisa Oviedo Glvez
................................ 56
Mancini J ...................... 94
Maria Blanco................. 87
Mara BoffIill Crdenas... 95
Mara de los ngeles
Bcquer.................... 61
Mara Elena Licea Surez
................................ 55
Mara Teresa Daz Soto 54,
55, 61, 62
Marian Hernndez
Colina ................... 132
Maribel Rivas ................ 87
Mariela Forrellat Barrios106
Mariela Guevara............ 73
Martha Hernndez......... 87
Martinez A .................... 52
Martnez Morales Lisbeth
.............................. 107
Martinez O.................... 52
Matthew S .................. 103
Maykel Prez Machn .... 95
ME Gonzlez Fraguela 119
Mercader OA .............. 125
Meza Cueto C.Y.......... 111
MH Rossi ...................... 21
Mollineda A................... 53
Montserrat Guardiola ..... 68
MR Kviecinski ............... 83
MS Farias............... 14, 42
Muoz-Montesino C.... 113,
114
N Brooks ...................... 36
Nadine Hempel ............. 22
Natalia M Inada........... 120
Nelson Merino............. 106
Nielsen Lagumersindez
Denis........................ 56
Nubia Fernndez
Hernndez........ 82, 121
O Marante Maldonado . 101
O Valds .................... 101
O Zerquera ................. 101
O. Ipcioglu ........ 37, 38, 39
O. Lara......................... 81
O. Sayan ...................... 39
O. Yildiz.................. 38, 39
Octavio Fernndez ........ 92
Olga Sonia Len Fernndez
.......................... 16, 77
Olinda Mayhuasca....... 109

Havana Redox 2009

Olivares S.....................94
Omaida Lpez Bernal ....95
Omar Ramrez Nez ...58,
66, 73
Oate S..............113, 114
Oscar Concepcin .........87
Ovando Cindy .............107
P Budni.............14, 40, 42
P Cos ...................10, 126
Pablo Bonilla...............109
Pandolfi A .50, 52, 89, 127
Pauline M. Carrico .........22
Pentn-Rol G ..........58, 66
Pea E .......................114
Pea E Sobarzo A.......113
Perde- Schrepler ...........23
Pereira N ..50, 52, 89, 127
Prez de Arma A...........53
Prez M........................53
Prez MR................97, 99
Prez, Y....................129
Peter J. O'Brien .............20
Pupo JB....50, 52, 89, 127
R Garca Miniet ...........119
R Geremias.......21, 44, 83
R Gonzlez...................88
R Or ...................80, 108
R Valdivieso................108
R. Garca-Bou ...............76
R. Gonzlez
.....................128
R. Ramrez....................81
R. Trujillo ....................104
R. Valdivieso.................80
Rayza Mndez Triana ....95
RC Pedrosa14, 21, 40, 42,
44, 83
Reinaldo Trujillo.............87
Ren Delgado Hernndez
................32, 105, 106
Ricardo Gonzlez........ 122
Richard Schulz.............. 33
Rivas C I..... 113, 114, 117
Rivera Castaeda E.G. 111
Rivern G ......... 50, 52, 89
Roberto Faure............... 92
Roblejo H ................. 127
Rodolfo Lpez..... 102, 110
Rodrguez CC ......... 97, 99
Rodriguez JL............... 125
Roger F. Castilho......... 105
Rosa Mara Flores Snchez
................................ 70
Rosendo A. Yunes......... 85
Rozangela C. Pedrosa... 85
Rubio Pino J.L............. 111
S vila Jnior ................ 40
S Ragapolan ................. 12
S Surez..................... 108
S. Celik............. 37, 38, 39
S. Nalbant......... 37, 38, 39
Saadath Hashamiyan..... 96
Santacruz, VC............... 78
Segundo M................. 103
Seoung Woo Shin........ 118
Silvia Daz Llera31, 88
Silvia Menndez.......... 122
Silvia Surez......... 80, 109
SJF Garcia.................... 83
Sobarzo A................... 114
Soliz C........................ 113
Sotomayor K....... 113, 114
SRS Ferreira................. 83
SS du Plessis................ 36
Stewart Sell................... 22
Sun Joo Lee................ 118
Sung Youl Kim ............ 118
T Horemans................ 126
T Nantembwa................ 64
T. Augustine................ 104
Teresa Montero........... 122
Tiberius Dicu................. 23
TMF Gnther .......... 44, 83
Tomasz Bilinski ............. 30
TR Garlet...................... 40
Trujillo R....................... 72
Tuncer OZEN........... 28
V Santacruz Vazquez .... 79
V. M. Vctor................... 76
Valenzuela-Silva CM58, 66
Vera JC...... 113, 114, 117
Victor Huerta Espinosa . 77,
78, 79, 90
Villagrn M.......... 113, 114
W Damin..................... 64
W.W. Lautt.................... 29
Y Alarcn.... 51, 57, 63, 65
Y Prez Torres.............. 88
Y Rubio .................. 51, 65
Y Saborit .................... 101
Y. Capdesuer ............ 104
Y. Garca .................. 128
Y. Gutirrez ................ 110
Y. Quirs .................... 104
Y. Tunca....................... 37
Yeny Lemus................ 106
YG Aboua..................... 36
Yolanda Valds
Rodrguez ............. 132
Yudelsy A. Tandrn ....... 87
Z. Radk....................... 26
Z. Yesilova........ 37, 38, 39
Zarate Maldonado Brenda
ngela...................... 77
Zullyt Zamora.............. 122
Ziga F............. 114, 117

Antioxidants

Mitochondrial damage in septic shock plus hypoxia: therapy with
mitochondria-targeted antioxidants

VM Vctor
a,b
, R Garca-Bou
c
, C Bauls
b
, JR Herance
d
, JV Esplugues
c
, A Hernndez-Mijares
a
, M Rocha
a,b

a
Service of Endocrinology. Universitary Hospital Dr. Peset. Avda. Gaspar Aguilar 90, 46017 Valencia. Spain.
b
Dr. Peset Hospital Research Foundation. Avda. Gaspar Aguilar 90, 46017 Valencia. Spain
c
Pharmacology Department and Ciberehd, Faculty of Medicine, University of Valencia C/ Blasco Ibaez 15, 46010 Valencia.
Spain.
d
Institute of High Technology The Barcelona Biomedical Research Centre, C/ Dr. Aiguader, 88 08033 Barcelona, Spain

Excessive production of nitric oxide (NO) contributes to the development of septic shock through its effects
on vascular tone. However, its inhibitory effect on mitochondrial respiration may play a role in the
pathophysiological mechanisms underlying organ dysfunction. Tissue hypoxia often coexists with sepsis in
clinical conditions. Therefore, we have evaluated the temporary effects of these modifications on the
inhibition of respiration as well as the influence of concurrent hypoxia. Human umbilical endothelial cells
(HUVECs) were activated by endotoxin, TNFalpha and IFNgamma over a 24-h period. Thereafter, cells
were incubated at 21% and 1.5% oxygen (normoxia and hypoxia respectively), and mitochondrial
respiration, complex I activity, peroxynitrite production, S-nitrosylation, antioxidant content and tyrosine
nitration were evaluated using the Clark oxygen electrode, fluorescent probes and western blot. Oxygen
consumption, complex I activity and antioxidant levels in activated cells fell progressively as time
progressed. This effect was largely prevented by coincubation with the non-specific N-nitro L arginine.
Addition of glutahione ester and mitoquinone, two mitochondrial antioxidants, reversed these effects at
early time points. Thereafter, the inhibition of complex I became more persistent, coinciding with a
progressive increase in nitration. Hypoxia accelerated the persistent inhibition of complex I, despite a
reduction in the total amount of NO generated. Our results suggest that hypoxia amplifies the
mitochondrial inhibition induced by the NO that is generated during inflammatory disease states such as
hypoxia, hypertension and atherogenesis, and endorse mitochondria-targeted antioxidants as a
therapeutic tool for these pathologies.

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Copyright DESOFT S.A. 2009 Revista Cubana de Farmacia Vol.

43 Supplement 1 2009, ISSN 0034-7515

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