Molecular Toxicology - practice

Data

Official data in SubjectManager for the following academic year: 2022-2023

Topic

In the practices, the students will carry out laboratory experiments involving methods used for in vitro toxicity testing. On prokaryotic and eukaryotic cells, including cells of animal and human origin, they examine the cyto- and genotoxic effects of xenobiotics of toxicological relevance (e.g. mycotoxins, metal ions, and enzyme inducers) and of UV radiation; they analyze the binding of chemicals (e.g. ochratoxin A) to human albumin, and the effects of some drugs on the expression of cytochrome P450 (CYP) genes. The students will be acquainted with the principles of the methods used, the mechanisms of action of the toxicants studied, the details of the experimental techniques, as well as the statistical analysis, presentation and interpretation of the experimental results. The laboratory work will be carried out in an interactive fashion. The students will perform the experiments jointly according to a detailed protocol previously provided for them.

Lectures

Practices

• 1. Investigation of protein-ligand interactions with fluorescence spectroscopy I
• 2. Investigation of protein-ligand interactions with fluorescence spectroscopy II
• 3. Investigation of protein-ligand interactions with fluorescence spectroscopy III
• 4. Investigation of protein-ligand interactions with fluorescence spectroscopy IV
• 5. Detection of UV-induced cyto- and genotoxicity in Serratia marcencens I
• 6. Detection of UV-induced cyto- and genotoxicity in Serratia marcencens II
• 7. Detection of UV-induced cyto- and genotoxicity in Serratia marcencens III
• 8. Detection of UV-induced cyto- and genotoxicity in Serratia marcencens IV
• 9. Detection of cytotoxicity by the Azotobacter agile assay I
• 10. Detection of cytotoxicity by the Azotobacter agile assay II
• 11. Detection of cytotoxicity by the Azotobacter agile assay III
• 12. Detection of cytotoxicity by the Azotobacter agile assay IV
• 13. Detection of genotoxicity in Chinese hamster ovary cells by comet assay I
• 14. Detection of genotoxicity in Chinese hamster ovary cells by comet assay II
• 15. Detection of genotoxicity in Chinese hamster ovary cells by comet assay III
• 16. Detection of genotoxicity in Chinese hamster ovary cells by comet assay IV
• 17. Assessment of the viability by ATP analysis of human HepG2 cells exposed to cytotoxic chemicals I
• 18. Assessment of the viability by ATP analysis of human HepG2 cells exposed to cytotoxic chemicals II
• 19. Assessment of the viability by ATP analysis of human HepG2 cells exposed to cytotoxic chemicals III
• 20. Assessment of the viability by ATP analysis of human HepG2 cells exposed to cytotoxic chemicals IV
• 21. Detection of CYP induction by drugs in HepG2 cells I
• 22. Detection of CYP induction by drugs in HepG2 cells II
• 23. Detection of CYP induction by drugs in HepG2 cells III
• 24. Detection of CYP induction by drugs in HepG2 cells IV
• 25. Detection of CYP induction by drugs in HepG2 cells V
• 26. Detection of CYP induction by drugs in HepG2 cells VI
• 27. Detection of CYP induction by drugs in HepG2 cells VII
• 28. Detection of CYP induction by drugs in HepG2 cells VIII

Seminars

Reading material

Obligatory literature

Literature developed by the Department

A detailed description of each practice (handout) will be available for the students on Neptun in advance of each practice. The handout will explain the principle of the methods, characterize the toxicants to be tested, describe the necessary reagents and eqipment, the steps of the experiments (protocol), the presentation, statistical analysis and evaluation of the the results. The students are expected to review the handout thoroughly before taking part on the practice.

Notes

Recommended literature

C.D. Klassen (ed.): Casarett & Doull's Toxicology. The Basic Science of Poisons. 9th edition, McGraww-Hill, 2019.

Conditions for acceptance of the semester

Maximum of 15 % absence allowed

Mid-term exams

The performance of students during laboratory practices will be evaluated and graded with a practical course mark, which will be based mainly on the quality of their written reports (laboratory notes) of each practice, however, their preparedness for the practices will also be considered.

Making up for missed classes

Missed classes cannot be made up.

Exam topics/questions

The performance of students during laboratory practices will be evaluated and graded with a practical course mark, which will be based mainly on the quality of their written reports (laboratory notes) of each practice, however, their preparedness for the practices will also be considered.

Examiners

Instructor / tutor of practices and seminars

• Dr. Lemli Beáta
• Hartnerné Dr. Pohóczky Krisztina