**Meet the editor**

Dr. Volodymyr I. Lushchak graduated from the Lomonosov Moscow State University (Soviet Union) in 1982. In 1986 he defended his Ph.D. Theisis and in 2002 his D.Sc. Dissertation. He obtained his full professor title in 2004. From 1987 to 1998 he worked in Southern Seas Biology Institute of Academy of Sciences of Ukraine. In 1998 Dr. Lushchak joined Vassyl Stefanyk Precarpathian Nation-

al University in Ivano-Frankivsk, Ukraine) where he currently works as the head of the Department of Biochemistry and Biotechnology. He also worked at Universities of Canada, Brazil, United Kingdom, Finland, Poland, Sweden and Germany. Dr. Lushchak's scientific interests are related to effects of environmental conditions and molecular aspects of adaptive responses in bacteria, fungi, plants and animals. Although interested in general aspects of metabolism regulation, his interests are mainly focused on the homeostasis of reactive species of oxygen, nitrogen and carbon.

Contents

**Preface IX** 

Chapter 1 **Introductory Chapter 3** 

**Section 2 Physical Factors 11** 

**Section 3 Chemical Factors 113** 

Chapter 6 **Oxidative Stress Induced** 

Volodymyr I. Lushchak

Chapter 2 **Oxidative Stress Induced Damage** 

Chapter 3 **Exercise and Oxidative Stress 33** 

**of the Human Retina: Overview** 

Vladimir Lj. Jakovljevic, Dejan Cubrilo,

**Stress Events in Antarctic Fungi 75**  Nedelina Kostadinova, Ekaterina Krumova,

Chapter 5 **Changes in Hydrogen Peroxide Levels and Catalase** 

Tayeb Wafa, Nakbi Amel, Chaieb Ikbal

and Hammami Mohamed

**Isoforms Expression are Induced With Freezing Tolerance by Abscisic Acid in Potato Microplants 99**  Martha E. Mora-Herrera, Humberto López-Delgado, Ernestina Valadez-Moctezuma and Ian M. Scott

**by the 2,4-Dichlorophenoxyacetic Herbicide 115** 

Chapter 4 **Transient Cold Shock Induces Oxidative** 

**of Mechanisms and Preventional Strategies 13**  Katrin Engelmann, Klio Ai Becker and Richard Funk

Vladimir Zivkovic, Dusica Djordjevic and Dragan Djuric

Tzvetanka Stefanova, Vladislava Dishliyska and Maria Angelova

**Section 1 Introduction 1** 

### Contents

#### **Preface XIII**

**Section 1 Introduction 1**  Chapter 1 **Introductory Chapter 3**  Volodymyr I. Lushchak **Section 2 Physical Factors 11**  Chapter 2 **Oxidative Stress Induced Damage of the Human Retina: Overview of Mechanisms and Preventional Strategies 13**  Katrin Engelmann, Klio Ai Becker and Richard Funk Chapter 3 **Exercise and Oxidative Stress 33**  Vladimir Lj. Jakovljevic, Dejan Cubrilo, Vladimir Zivkovic, Dusica Djordjevic and Dragan Djuric Chapter 4 **Transient Cold Shock Induces Oxidative Stress Events in Antarctic Fungi 75**  Nedelina Kostadinova, Ekaterina Krumova, Tzvetanka Stefanova, Vladislava Dishliyska and Maria Angelova Chapter 5 **Changes in Hydrogen Peroxide Levels and Catalase Isoforms Expression are Induced With Freezing Tolerance by Abscisic Acid in Potato Microplants 99**  Martha E. Mora-Herrera, Humberto López-Delgado, Ernestina Valadez-Moctezuma and Ian M. Scott **Section 3 Chemical Factors 113** 

Chapter 6 **Oxidative Stress Induced by the 2,4-Dichlorophenoxyacetic Herbicide 115**  Tayeb Wafa, Nakbi Amel, Chaieb Ikbal and Hammami Mohamed


Contents VII

Chapter 17 **Effects of NK-4, a Cyanine Dye** 

**with Antioxidant Activities: Attenuation of Neuronal Deficits in Animal Models of Oxidative Stress-Mediated Brain** 

**Ischemia and Neurodegenerative Diseases 369** Hitomi Ohta, Kenji Akita and Tsunetaka Ohta


#### **Section 5 Antioxidants 201**


Chapter 17 **Effects of NK-4, a Cyanine Dye with Antioxidant Activities: Attenuation of Neuronal Deficits in Animal Models of Oxidative Stress-Mediated Brain Ischemia and Neurodegenerative Diseases 369**  Hitomi Ohta, Kenji Akita and Tsunetaka Ohta

VI Contents

Chapter 7 **Environmental Pollution and Oxidative Stress in Fish 131** Oksana B. Stoliar and Volodymyr I. Lushchak

**in Citrus by** *Xanthomonas citri* **pv** *citri* **169**

Tiiu Kullisaar, Epp Songisepp and Mihkel Zilmer

Rajagopalan Vijayaraghavan and Anshoo Gautam

**on Secretory Function in Salivary Gland Cells 189**

**Section 4 Biological Factors and Effects 167**

Chapter 9 **Effect of Oxidative Stress** 

**Section 5 Antioxidants 201**

Chapter 12 **Flavonoid Treatment** 

Chapter 8 **Interference of Oxidative Metabolism** 

Robert C. Ebel and Naveen Kumar

Ken Okabayashi, Takanori Narita, Yu Takahashi and Hiroshi Sugiya

Chapter 10 **Probiotics and Oxidative Stress 203**

Chapter 13 **The Effects of Propolis in Animals** 

Chapter 14 **Antioxidants in Thai Herb, Vegetable** 

Warin Sangkitikomol

Fahmy G. Elsaid

Chapter 16 **Dietary Antioxidants:** 

Chapter 15 **Modification by Aqueous Extracts** 

Chapter 11 **Diabetes, Oxidative Stress and Tea 223** B. Alipoor, A. Homayouni Rad and E. Vaghef Mehrabany

**for Mustard Agents' Toxicity 249**

**Exposed Oxidative Stress 267** Pinar Tatli Seven, Seval Yilmaz,

Ismail Seven and Gulizar Tuna Kelestemur

**and Fruit Inhibit Hemolysis and Heinz**

**Body Formation in Human Erythrocytes 289** 

**of** *Allium kurrat L.* **and** *Ricinus communis L.* **of Cyanide Nephrotoxicity on Balb/C Mice 307**

**From Micronutrients and Phytochemicals** 

**on Early Atherosclerosis and Obesity 323**

**to Enzymes – Preventive Effects**

Sylvie Gaillet, Dominique Lacan

and Jean-Max Rouanet

Preface

Free radicals discovered in biological systems in 1950es were immediately suggested to be involved in diseases and aging (Harman, 1956; 1985). The term "free radicals" was later extended to denote a wider group of activated oxygen forms whose activity is higher than molecular oxygen, and were collectively named reactive oxygen species (ROS), which include singlet oxygen, superoxide anion radical, hydrogen peroxide, hydroxyl radical, and many of their derivatives. In 1969, J. McCord and I. Fridovich described the catalytic function for erythrocuprein (hemocuprein) as superoxide dismutase responsible for elimination of the superoxide anion. The information on free radical processes in biological systems allowed Helmut Sies (1985) to systematize "Oxidative stress" and came to denote a disturbance in the prooxidant-antioxidant balance in favor of the former. Recently, we modified this definition as "Oxidative stress is a situation when steady-state ROS concentration is transiently or chronically enhanced, disturbing cellular metabolism and its regulation, and damaging cellular constituents" (Lushchak, 2011b). The last definition included accumulated the up-todate knowledge on the effects of ROS on core and regulatory processes, and underlined the idea on their steady-state level in biological systems. Our understanding of the ROS roles in biological systems has gone through three phases: their appreciation as damaging ones, protection against infections and, finally, signaling and regulatory molecules in diverse biological processes. We can now state that all listed components operate in organisms in concert and are absolutely

Intensive research was invested into discovering whether the environmental factors can affect intracellular ROS steady-state levels. That resulted in understanding that this level may be modified by many external physical, chemical and biological factors. Since it is difficult to register ROS levels *in situ*, these data were mainly gained through indirect methods with the evaluation of levels of ROS-modified molecules of both external and internal origin. Therefore, this book mainly contains the information on oxidative stress induced by physical and chemical factors and a portion of the book

On January 2, 2012, a Google Scholar search for "oxidative stress environment" yielded about 589,000 publication hits, whereas in Scopus and Pubmed databases it yielded 4,428 and 6,302 hits, respectively. We have presented 17 chapters in this book,

includes the information on antioxidants capable to modify ROS levels.

necessary for realization of biological functions.

## Preface

Free radicals discovered in biological systems in 1950es were immediately suggested to be involved in diseases and aging (Harman, 1956; 1985). The term "free radicals" was later extended to denote a wider group of activated oxygen forms whose activity is higher than molecular oxygen, and were collectively named reactive oxygen species (ROS), which include singlet oxygen, superoxide anion radical, hydrogen peroxide, hydroxyl radical, and many of their derivatives. In 1969, J. McCord and I. Fridovich described the catalytic function for erythrocuprein (hemocuprein) as superoxide dismutase responsible for elimination of the superoxide anion. The information on free radical processes in biological systems allowed Helmut Sies (1985) to systematize "Oxidative stress" and came to denote a disturbance in the prooxidant-antioxidant balance in favor of the former. Recently, we modified this definition as "Oxidative stress is a situation when steady-state ROS concentration is transiently or chronically enhanced, disturbing cellular metabolism and its regulation, and damaging cellular constituents" (Lushchak, 2011b). The last definition included accumulated the up-todate knowledge on the effects of ROS on core and regulatory processes, and underlined the idea on their steady-state level in biological systems. Our understanding of the ROS roles in biological systems has gone through three phases: their appreciation as damaging ones, protection against infections and, finally, signaling and regulatory molecules in diverse biological processes. We can now state that all listed components operate in organisms in concert and are absolutely necessary for realization of biological functions.

Intensive research was invested into discovering whether the environmental factors can affect intracellular ROS steady-state levels. That resulted in understanding that this level may be modified by many external physical, chemical and biological factors. Since it is difficult to register ROS levels *in situ*, these data were mainly gained through indirect methods with the evaluation of levels of ROS-modified molecules of both external and internal origin. Therefore, this book mainly contains the information on oxidative stress induced by physical and chemical factors and a portion of the book includes the information on antioxidants capable to modify ROS levels.

On January 2, 2012, a Google Scholar search for "oxidative stress environment" yielded about 589,000 publication hits, whereas in Scopus and Pubmed databases it yielded 4,428 and 6,302 hits, respectively. We have presented 17 chapters in this book, covering several important aspects of environmentally induced oxidative stress and its prevention by antioxidants. Since oxidative stress seems to be an inevitable component of virtually all stresses that are strong enough, the book provides the interested readers with information needed to recognize this.

Preface XI

The next section is devoted to induction of oxidative stress by biological factors. Diverse pathogens invading the host organism are attacked by the immune system equipped by machinery to produce reactive species. R. C. Ebel and N. Kumar investigated the involvement of reactive oxygen species in combating *Xanthomonas citri pv citri* (Xcc), causing citrus canker in *Citrus sp*. and found that pathogen-induced oxidative stress was differently expressed in different representatives of the genera studied. K. Okabayashi et al. were able to demonstrate that ethacrynic acid, a thiolmodulating reagent, inhibited amylase release induced by β-adrenergic agonist in rat parotid acinar cells and the effect was independent of depletion of glutathione in the cells. The authors concluded that the inhibitory effect of ethacrynic acid on amylase release induced by β-adrenergic agonist was caused by the thiol-modulation of β-

It is very attractive to use antioxidants to prevent ROS-induced modification of organisms' functions. Intuitively developed at the beginning of ROS investigation in living organisms, it looked promising to use them for prophylactics and treatment of ROS-modulated damages. However, the promises were not realized and it became clear that there are no absolutely direct links between ROS-induced changes and pathologies. The last section of the book presents a broad discussion of positive effects of diverse antioxidants. The Estonian team led by T. Kullisaar provides an interesting topic – after short surveys on probiotics and oxidative stress they share extensive information on the potential use of different probiotics in functional foods and capsules that may be helpful to combat oxidative stress related to many pathologies, like cardiovascular diseases, metabolic syndrome, allergy, atopic dermatitis, radiationinduced problems in the intestinal tract. Diabetes is a very common human disease, which, in addition to health problems caused, is accompanied by many complications related with oxidative stress and the system character of the pathology therefore clearly needs specific approaches. It is very attractive to use a food stuff instead drugs and B. Alipoor et al. describe the potential of one of the most common drinks, tea, with health benefits particularly for diabetes and related complications. Sulphur mustard as a bifunctional alkylating agent readily reacts with a variety of macromolecules including nucleic acids, proteins and lipids, as well as small molecular mass metabolites such as glutathione, which is in the focus of chapter written by R. Vijayaraghavan and A. Gautam. Since sulphur mustard also induces oxidative stress, antioxidants can be useful and the authors analyze available data on the use of flavonoids, particularly from *Hippophae rhamnoides.* Bee products accompanied people since ancient times and only now do we start to understand the molecular mechanisms of many processes modulated by these products. Therefore, P. Tatli Seven provide an extensive analysis of beneficial properties of propolis with the focus on its antioxidant, antimicrobial, anti-inflammatory and antitumor effects. The antioxidant potential of 152 samples of Thai fruits, vegetables and herbs, and 33 brands of tea was measured by W. Sangkitikomol and this study shows that the products are a good source of compounds with health benefits. Since the toxicity of cyanide is associated with the induction of oxidative stress, F. G. Elsaid suggests and proves that it can be

adrenergic receptors.

The Introduction section (V. I. Lushchak) covers general aspects of oxidative stress theory and briefly analyses potential ways of oxidative stress induction by environmental factors – stimulation of ROS production and depletion of antioxidants. The role of antioxidants is also highlighted.

The book is divided into four parts. The first section, entitled "Physical Factors" demonstrates the induction of oxidative stress by exercise, light and temperature fluctuations. The chapter written by V. Lj. Jakovljevic and colleagues extensively introduces the biology of reactive oxygen and nitrogen species, measurement of redox status, levels of superoxide anion radical, hydrogen peroxide, glutathione, lipid peroxides, activities of superoxide dismutase and catalase, and then demonstrates that exercise may increase the production of ROS and modify redox status. Interestingly, it has been demonstrated that perturbations of free radical processes depend on the intensity and type of exercise, as well as specialization of athletes and their physical state. Different light types possessing high energy can also induce damage to cellular components, even in specialized organs. K. Engelmann et al. described the operation of human retina, ROS-related processes, protective role of specific parts of the light spectrum and retina protection by tinted intraocular lenses in detail. The next two experimental chapters deal with oxidative stress induced by temperature changes – in fungi and plants. Using two Antarctic fungi, *Penicillium sp.* and *Aspergillus glaucus*, N. Kostadinova et al. demonstrated a relationship between cold shock and oxidative stress evidenced by an increased level of oxidized proteins and activation of antioxidant enzymes. Since abscisic acid may increase freezing tolerance of plants, M. E. Mora-Herrera et al. were able to demonstrate that ther decrease in temperature affected the level of hydrogen peroxide and catalase isoforms in potato microplants, which was related to tolerance to low temperatures.

The induction of oxidative stress by chemical factors is presented in the second section of the book. Ions of metals may induce oxidative stress in at least two ways – entering Fenton reaction and replacing other metal ions in their binding centers (Valko et al., 2007). The detailed description of toxicokinetics of lead and cadmium, induction and role of oxidative stress in neurochemical changes in the hypothalamus and pituitary of F1 generation PND 56 male and female rats are presented by P. Pillai et al. Herbicides are well known inducers of oxidative stress and many mechanisms were described in this case. 2,4-Dichlorophenoxyacetic herbicide is one of the broadly used ones, and W. Tayeb et al. describe the general phenomenology and potential mechanisms of induction of oxidative stress in different organisms. The chapter by O. B. Stoliar and V. I. Lushchak is devoted to analysis of oxidative stress induced in fish by different environmental pollutants.

The next section is devoted to induction of oxidative stress by biological factors. Diverse pathogens invading the host organism are attacked by the immune system equipped by machinery to produce reactive species. R. C. Ebel and N. Kumar investigated the involvement of reactive oxygen species in combating *Xanthomonas citri pv citri* (Xcc), causing citrus canker in *Citrus sp*. and found that pathogen-induced oxidative stress was differently expressed in different representatives of the genera studied. K. Okabayashi et al. were able to demonstrate that ethacrynic acid, a thiolmodulating reagent, inhibited amylase release induced by β-adrenergic agonist in rat parotid acinar cells and the effect was independent of depletion of glutathione in the cells. The authors concluded that the inhibitory effect of ethacrynic acid on amylase release induced by β-adrenergic agonist was caused by the thiol-modulation of βadrenergic receptors.

X Preface

covering several important aspects of environmentally induced oxidative stress and its prevention by antioxidants. Since oxidative stress seems to be an inevitable component of virtually all stresses that are strong enough, the book provides the interested

The Introduction section (V. I. Lushchak) covers general aspects of oxidative stress theory and briefly analyses potential ways of oxidative stress induction by environmental factors – stimulation of ROS production and depletion of antioxidants.

The book is divided into four parts. The first section, entitled "Physical Factors" demonstrates the induction of oxidative stress by exercise, light and temperature fluctuations. The chapter written by V. Lj. Jakovljevic and colleagues extensively introduces the biology of reactive oxygen and nitrogen species, measurement of redox status, levels of superoxide anion radical, hydrogen peroxide, glutathione, lipid peroxides, activities of superoxide dismutase and catalase, and then demonstrates that exercise may increase the production of ROS and modify redox status. Interestingly, it has been demonstrated that perturbations of free radical processes depend on the intensity and type of exercise, as well as specialization of athletes and their physical state. Different light types possessing high energy can also induce damage to cellular components, even in specialized organs. K. Engelmann et al. described the operation of human retina, ROS-related processes, protective role of specific parts of the light spectrum and retina protection by tinted intraocular lenses in detail. The next two experimental chapters deal with oxidative stress induced by temperature changes – in fungi and plants. Using two Antarctic fungi, *Penicillium sp.* and *Aspergillus glaucus*, N. Kostadinova et al. demonstrated a relationship between cold shock and oxidative stress evidenced by an increased level of oxidized proteins and activation of antioxidant enzymes. Since abscisic acid may increase freezing tolerance of plants, M. E. Mora-Herrera et al. were able to demonstrate that ther decrease in temperature affected the level of hydrogen peroxide and catalase isoforms in potato microplants,

The induction of oxidative stress by chemical factors is presented in the second section of the book. Ions of metals may induce oxidative stress in at least two ways – entering Fenton reaction and replacing other metal ions in their binding centers (Valko et al., 2007). The detailed description of toxicokinetics of lead and cadmium, induction and role of oxidative stress in neurochemical changes in the hypothalamus and pituitary of F1 generation PND 56 male and female rats are presented by P. Pillai et al. Herbicides are well known inducers of oxidative stress and many mechanisms were described in this case. 2,4-Dichlorophenoxyacetic herbicide is one of the broadly used ones, and W. Tayeb et al. describe the general phenomenology and potential mechanisms of induction of oxidative stress in different organisms. The chapter by O. B. Stoliar and V. I. Lushchak is devoted to analysis of oxidative stress induced in fish by different

readers with information needed to recognize this.

which was related to tolerance to low temperatures.

environmental pollutants.

The role of antioxidants is also highlighted.

It is very attractive to use antioxidants to prevent ROS-induced modification of organisms' functions. Intuitively developed at the beginning of ROS investigation in living organisms, it looked promising to use them for prophylactics and treatment of ROS-modulated damages. However, the promises were not realized and it became clear that there are no absolutely direct links between ROS-induced changes and pathologies. The last section of the book presents a broad discussion of positive effects of diverse antioxidants. The Estonian team led by T. Kullisaar provides an interesting topic – after short surveys on probiotics and oxidative stress they share extensive information on the potential use of different probiotics in functional foods and capsules that may be helpful to combat oxidative stress related to many pathologies, like cardiovascular diseases, metabolic syndrome, allergy, atopic dermatitis, radiationinduced problems in the intestinal tract. Diabetes is a very common human disease, which, in addition to health problems caused, is accompanied by many complications related with oxidative stress and the system character of the pathology therefore clearly needs specific approaches. It is very attractive to use a food stuff instead drugs and B. Alipoor et al. describe the potential of one of the most common drinks, tea, with health benefits particularly for diabetes and related complications. Sulphur mustard as a bifunctional alkylating agent readily reacts with a variety of macromolecules including nucleic acids, proteins and lipids, as well as small molecular mass metabolites such as glutathione, which is in the focus of chapter written by R. Vijayaraghavan and A. Gautam. Since sulphur mustard also induces oxidative stress, antioxidants can be useful and the authors analyze available data on the use of flavonoids, particularly from *Hippophae rhamnoides.* Bee products accompanied people since ancient times and only now do we start to understand the molecular mechanisms of many processes modulated by these products. Therefore, P. Tatli Seven provide an extensive analysis of beneficial properties of propolis with the focus on its antioxidant, antimicrobial, anti-inflammatory and antitumor effects. The antioxidant potential of 152 samples of Thai fruits, vegetables and herbs, and 33 brands of tea was measured by W. Sangkitikomol and this study shows that the products are a good source of compounds with health benefits. Since the toxicity of cyanide is associated with the induction of oxidative stress, F. G. Elsaid suggests and proves that it can be reduced by the application of aqueous extracts of *Allium kurrat* and *Ricinus communis* which possess antioxidant properties. Due to high sugar and fat diets and sedentary lifestyles, modern people are frequently subjected to atherosclerosis and obesity, which are important risk factors for metabolic syndrome and greatly predispose individuals to liver diseases, cardiovascular disease, type 2 diabetes, dyslipidemia, hypertension and numerous cancers, and is associated with markedly diminished life expectancy. The French team (S. Gaillet, D. Lacan, J.-M. Rouanet) presents results of titanic systematic work to identify the beneficial diets and find a broad set of diary foods and beverages possessing antioxidant properties and helping to combat the mentioned pathologies. These products are fresh and possessed fruits grapes, and berries, preparations from them as well as selenium-enriched microalgae, algal and fungal polysaccharides. Recently, while screening more than 250 cyanine dyes for their neurotrophin-like activity, the compound called NK-4 and some related compounds were found to be potent neurotrophic agents for the promotion of growth and differentiation of neuronal rat adrenal pheochromocytoma cell line PC12. NK-4 is a divalent cationic pentamethine trinuclear cyanine dye that contains three quinolinium rings, N-alkyl side chains, and two iodine anions. In the last chapter of the book, the Japanese team (H. Ohta, K. Akita & T. Ohta) summarized the data on the biological effects in different models and found that NK-4 possesses free radical-scavenging activity, neuroprotective against various cytotoxic stresses, neuroprotective effects against β-amyloid (Aβ) toxicity, and intracellular signaling. Therefore, the authors suggest that this dye can be used to protect animal organisms against neurodegeneration**.**

This book is expected to be interesting to experts in the field of basic investigations of reactive oxygen species and oxidative stress, as well as to practical users in the diverse fields like environmental sciences, medicine, and toxicology.

> **Prof. Dr. Volodymyr I. Lushchak**  PhD, DSc, Department of Biochemistry and Biotechnology, Vassyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine
