CURRICULUM OF THE LECTURES 1. Protein and amino acid degradation. Proteolysis, proteolytic enzymes, proteasome. Amino acid degradation (transamination, oxidative deamination). Glucogenic and ketogenic amino acids. 2. Urea cycle. Amino acid decarboxylation, synthesis of biogenic amines. Biosynthesis of non-essential amino acids. Essential, non-essential amino acids, limiting amino acid. Biosynthesis of glutamate, serine, aspartate and cysteine families. Glutathione – γ-glutamyl cycle. 3. Nucleotide metabolism. Biosynthesis of purine nucleotides. Biosynthesis of pyrimidine nucleotides. Synthesis of deoxyribonucleotides. ATP – a substrate for the synthesis of NAD+, FAD and coenzyme A. Degradation of purines. Degradation of pyrimidines. 4. DNA replication and transcription. Replicons. DNA replication mechanism in prokaryotic and eukaryotic cells. Principle of DNA transcription. Promotors. Termination of RNA synthesis. Posttranscriptional modification of eukaryotic RNA. 5. Proteosynthesis - translation. Amino acid activation before proteosynthesis. tRNA modification. Three phases of proteosynthesis (initiation, elongation, termination). Ribosomal synthesis of proteins in prokaryotes and eukaryotes. Gene expression regulation. 6. Modification and maturation of native proteins. Protein sorting. Secretory and cytoplasmic pathway. Translocation signals, glycosylation, exocytosis. Protein folding, chaperons and chaperonins. 7. Biochemistry of digestion. Nutrients, organic substances, vitamins, minerals and trace elements. Hydrolysis and resorption of food components. Digestive juices. Digestion and resorption. 8. Specific metabolism of ruminants. Saccharide and lipid metabolism of rumen. Nitrogen metabolism. 9. Biochemistry of liver. Functions of the liver. Hepatic metabolism: carbohydrate, lipid (bile acids and bile salts), amino acid and protein metabolism, biotransformations, storage. Ethanol metabolism. 10. Biochemistry of blood. Blood: composition and functions. Plasma proteins, lipoproteins. Heme biosynthesis and heme degradation. Hemoglobin, iron metabolism, gas transport, erythrocyte metabolism. Acid-base balance. 11. Biochemistry of kidneys and acid-base balance. Functions of the kidneys. Urine formation: ultrafiltration, resorption, secretion, clearance. Organic and inorganic components of urine. Functions in the acid-base balance: proton and ammonia excretion. Electrolyte and water recycling: calcium and phosphate ions, sodium ions, water. Renal hormones: calcitriol, erythropoietin, renin-angiotensin system. 12. Biochemistry of muscle. Mechanism of muscle contraction. Regulation of muscle contraction. Energy metabolism in the white and red muscle fibers, creatine metabolism. Cori and alanine cycle. Biochemistry of connective tissue. Bone and teeth. Calcium metabolism. Collagens. Extracellular matrix. 13. Biochemistry of nerve system Signal transmission in the CNS, synaptic signal transmission. Resting and action potential. Neurotransmitters: acetylcholine, biogenic amines, peptides, purine derivatives, catecholamines. Receptors for neurotransmitters (ionotropic, metabotropic), acetylcholine receptors (nicotinic, muscarinic). Sight. SCHEME OF THE PRACTICAL LESSONS 1. Amino acid reactions. a) Ninhydrin reaction for the detection of free amino group. b) Xanthoprotein reaction for the detection of aromatic amino acids. c) Pettenkoffer reaction for the detection of tryptophan. d) Adamkiewicz reaction for the detection of tryptophan. e) Pauly reaction for the detection of histidine and tyrosine. f) The detection of thiol groups in amino acids containing sulphur. 2. Protein reactions. a) Determination of the isoelectric point of albumin. b) Protein precipitation by boiling. c) Protein precipitation by organic acids.. d) Protein precipitation by heavy metal salts. e) Salting out of serum proteins. f) Protein detection by biuret test. 3. Quantitative determination of proteins. a) The Bradford method for quantitative determination of proteins. b) The Lowry method for quantitative determination of proteins. c) Determination of total proteins in blood serum of animals by biuret method. d) Determination of albumin concentration in blood serum of animals. 4. 1st written credit test: Lipid and cholesterol metabolism. Protein and amino acid catabolism. Serum/plasma proteins of animals. a) Electrophoretic separation of plasma proteins (tutorial). b) Turbidimetric determination of immunoglobulins in blood serum c) Protein dialysis. 5. Metabolism of nucleic acids. a) DNA isolation. b) Polymerase chain reaction. c) Detection of PCR product using gel electrophoresis. 6. Amino acid nitrogen metabolism – transamination reactions. Determination of aspartate aminotransferase (AST, EC 2.6.1.1) activity in blood serum of animals. 7. Biochemistry of liver. a) Determination of alanine aminotransferase (ALT, EC 2.6.1.2) activity in blood serum of animals. b) Determination of γ-glutamyltransferase (GGT, EC 2.3.2.2) activity in blood serum of animals. 8. Mineral metabolism. a) Determination of calcium concentration in blood serum of animals. b) Determination of magnesium concentration in blood serum of animals. 9. Biochemistry of blood. a) Determination of hemoglobin concentration in blood of cattle. b) Determination of bilirubin concentration in blood serum of animals. c) Determination of iron concentration in blood serum of animals. 10. Biochemistry of kidney. a) Determination of creatinine concentration in blood serum of animals. b) Determination of urea concentration in blood serum of animals. 11. Biochemistry of muscle contraction. Determination of creatine kinase (CK, EC 2.7.3.2.) activity in blood serum of animals. 12. 2nd written test: Organ biochemistry. Biochemistry of bone. Determination of alkaline phosphatase (ALP, EC 3.1.3.1) activity in blood serum of animals. 13. Immunochromatographic methods. Detection of human chorionic gonadotropin (hCG) in urine (early pregnancy test, baby test). Student presentations of their seminary works. Awarding credit points to students. |