Redoxbiology

Faculty of Biology

Subject: Redoxbiology

Lecturer: Adil Bayzhumanov, Ph.D.

Course Objective As a result of mastering the discipline, the students will receive basic knowledge about oxidative stress and the mechanisms of antioxidant defense of cells, and get acquainted with biochemical and biophysical methods for assessing oxidative stress and the antioxidant status of cells.

Brief description of the academic discipline Redoxbiology is a special course during which students will acquire knowledge about the role of molecular oxygen in the life and metabolism of the cell, as well as the stages of cellular respiration (glycolysis, the Krebs cycle, and oxidative phosphorylation), about the main mechanisms for the formation of reactive oxygen species in the cell, the regulation of the system antioxidant protection and methods for assessing oxidative damage, the number of reactive oxygen species and the antioxidant status of the body.

During the seminars, students will get acquainted with the characteristics of the main reactive oxygen species formed in the cell, getting the knowledge what is oxidative stress and oxidative damage. They will receive information about various human diseases, in the pathogenesis of which oxidative stress plays a leading role, study methods for diagnosing oxidative stress and the characteristics of the antioxidant system, gain knowledge about the main enzymes of the antioxidant system and non-enzymatic antioxidants, as well as the molecular mechanisms of regulation of the Keap1/Nrf2/ARE signaling pathway and possible ways of modifying ferroptosis.

Topics of classes (lectures, seminars, projects) Lesson topics

Topic 1. Introduction to biochemistry and redox biology.

The elements that make up the body. Main classes of biomolecules. Redox reactions. (4 hours)

Topic 2. Cellular respiration.

Oxygen catastrophe. Archaea, prokaryotes, and eukaryotes, differences in the structure and composition of membranes. Stages of cellular respiration: glycolysis, pyruvate decarboxylation, Krebs cycle, and oxidative phosphorylation. (8 hours)

Topic 3. Reactive oxygen species (ROS)

Properties of molecular oxygen. Characteristics and classification of ROS. Singlet oxygen. Superoxide anion radical. Hydrogen peroxide. Nitrogen oxide. Mechanisms of ROS formation in physiological processes. Enzymatic synthesis of nitric oxide and inhibitors of this synthesis. ROS as signaling molecules. (6 hours)

Topic 4. Oxidative stress and its role in pathological processes.

Definition of oxidative stress. Formation of ROS upon irradiation with ionizing radiation. Radiolysis of water. Lipid peroxidation. Photodynamic therapy. Examples of diseases associated with oxidative stress. Free radical theory of aging - arguments for and against. (4 hours)

Topic 5. Non-enzymatic antioxidants

Fat-soluble and water-soluble antioxidants. Mechanisms of action of non-enzymatic antioxidants. reduced glutathione. Chelators of metals of variable valence. Vitamin C. Carnosine. Tocopherol. Polyphenols. Carotenoids. Sources of natural antioxidants. Assessment of total antioxidant activity. (2 hours)

Topic 6. Main antioxidant enzymes.

Superoxide dismutase. Catalase. Glutathione peroxidase. Mechanisms of their action. Examples of methods for determining the activity of antioxidant enzymes. Mutations in the genes of antioxidant defense enzymes, for example, amyotrophic lateral sclerosis and mutations in the SOD1 gene. Acatalasia. Ceruloplasmin. (4 hours)

Topic 7. Signaling pathways for the regulation of the antioxidant system. Regulation of the Keap1/Nrf2/ARE signaling pathway. Differences in the Keap1/Nrf2/ARE signaling pathway in the naked mole rat. Transcription factors of the FoxO family. The role of ROS in the development of the mitochondrial signaling pathway of apoptosis. Ferroptosis - what is the difference from other types of necrosis. ferroptosis inductors. (4 hours)

Topic 8. Markers of oxidative stress and damage, methods for their registration

Oxidative damage. Determination of ROS using the spin probe method. Measurement of hydrogen peroxide in cells and biological fluids and tissues. Measurement of "fingerprints" of ROS. Markers of lipid peroxidation. 8-hydroxy-20-deoxyguanosine (8OHdG) and 8-nitroguanine as markers of oxidative damage to nucleic acids. Markers of oxidative damage to proteins. Chemiluminescent method for the determination of ROS. Fluorescent dye methods for determining ROS in cells. Using nanotechnology to determine ROS and oxidative damage. Advantages and disadvantages of methods. (2 hours)

Duration of the academic discipline (course) 9 weeks

34 hours of lectures

36 hours of self-work

Reporting form Final assessment

Maximum number of points/percentages 100 points (100%)

Final grade structure (how many percent/points of the final grade is taken up by each type of student work) • Final certification 40 (40%)

• Attendance at the course is compulsory

• Active work in the classroom 15 (15%)

• Interim controls 15 (15%)

• Homework/self-work 15 (15%)

• Other - preparation of reports at seminars _ 15...(15%)

How many classes can be missed without a valid excuse? 0

List of recommended reading 1. Lehninger Principles of Biochemistry, Eighth Edition| ©2021 David L. Nelson; Michael M. Cox

2. Andrey B. Rubin. Fundamentals of Biophysics. Wiley–Scrivener Publishing, Hoboken–Salem, 2014

Additional materials and recommended Internet resources 1. Banerjee R., Becker D., Dickman M., Gladyshev V., Ragsdale S. Redox Biochemistry. Hoboken, New Jersey: John Wiley & Sons, 2008. 

2. Beatriz Alvarez, Marcelo Comini, Gustavo Salinas, Madia Trujillo Redox Chemistry and Biology of Thiols. 1st Edition - May 25, 2022

Paperback ISBN: 9780323902199 eBook ISBN: 9780323915663

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