Laboratory Manual for Practical Biochemistry

Description

Ministry of Healthcare of the Russian Federation theme 1.1. amino acid structUre and classification. Protein structure INTRODUCTION SELF-STUDY QUESTIONS Practical COLOR QUALITATIVE PROTEIN AND AMINO ACID REACTIONS Reagents Material of the investigation NINHYDRIN TEST Procedure and observation: Principle TESTS FOR SULPHUR CONTAINING AMINO ACIDS AND PROTEINS Principle Procedure & observation Practical use of color reactions theme 1.2. sTRUCTURE, PHYSiCAL AND CHEMICAL PROPERTIES OF PROTEINS iNTRODUCTION SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical 1 Reagents Material of investigation Principle Design of laboratory work Practical 2 PROTEIN DENATURATION Reagents Material forthe investigation Principle Chemical denaturation. Practical significance Design of laboratory work theme 1.3. protein classification. protein structure and functions in human body. complex proteins introduction SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical Reagents Analysis of phosphoprotein chemical composition Material of the investigation Analysis of glycoprotein chemical composition Material for the investigation Procedure and observation Design of practical Principle Principle Principle Procedure Principle Procedure Tests case studies theme 2.1. FAT SOLUBLE VITAMINS INTRODUCTION SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical 1 QUAliTATIVE RECTIONS OF FAT SOLUBLE VITAMINS RETINOL QUALIITATIVE REACTION Principle Reagents Material for investigation Procedure and observation CHOLECALCIFEROL QUALITATIVE REACTION Principle Reagents Material for investigation Procedure and observation TOCOFEROL QUALITATIVE REACTION Principle Reagents Material for investigation Procedure and observation VITAMIN K QUALITATIVE REACTION Principle Menadione (synthetic analogue of vitamin K1) in the presence of cysteine in a basic medium turns to lemon yellow color. Reagents Material for investigation Procedure and observation Clinical and diagnostic significance Design of laboratory work theme 2.2. WATER SOLUBLE VITAMINS INTRODUCTION SELF-STUDY QUESTIONS topics for report Practical quaLItative reactions OF water soluble vitamins Reagents Equipment Material for investigation riboflavin qualitative reaction Material for investigation Reducing reaction Principle Procedure and observation nicotinic acid qualitative reaction Principle Material for investigation Procedure and observation pyridoxine qualitatuvereaction Principle Material for investigation Procedure and observation cobalamin qualitative reaction Principle Material for investigation Procedure and observation ascorbic acid qualitative reaction Material for investigation Principle Procedure and observation Design of laboratory work tests case studies Theme 3.1. ENZYME STRUCTURE AND properties. APPLICATION IN MEDICINE INTrODUCTION the aim of practical is SELF-STUDY QUESTIONS topic for reports Practical 1 dependence of enzyme activity on temperature Investigation of influence of temperature on salivary amylase activity Reagents Material of investigation Principle Procedure and observation. Preparation of saliva (the student on duty performs the saliva for the whole group). Design of laboratory work Practical 2 Investigation of enzyme specificity Reagents Material for investigation Detection of urease absolute specificity Principle Procedure Urease preparation: Design of laboratory work Detection of salivary amylase specificity Principle Procedure and observation Preparation of saliva (the student on duty performs the saliva for the whole group). Design of laboratory work theme 3.2. ENZYME ACTIVITY REGULATION INTRoDUCTION THE AIM OF THE PRACTICAL IS SELF-STUDY QUESTIONS Practical DETERMINATION OF AMYLASE ACTIVITY IN SERUM and urine Principle Reagents Material for investigation Procedure Calculation: Normal values Clinical and diagnostic significance Serum and urine Design of laboratory work Theme 3.3. enzyme classification and nomenclature (seminar) introduction the aim of the seminar SELF-STUDY QUESTIONS topic for reports tests case studies Theme 4.1. main catabolic pathways: pyruvate oxidative decarboxylation. tricarbonic acid cycle. enzymes of respiratory chain. oxidaTive phosphorylation (seminar) introduction the aim Of practic is SELF-STUDY QUESTIONS TESTS Case studies theme 5.1. externalmetabolism of PROTEINs. PRoteinsDIGESTION AND ABSORPTION introduction the aim of THE practicAL CLASS is SELF-STUDY QUESTIONS topics for reports Practical 1 qualitative reaction of free hyDrochloric acid in gastric juice Material of investigation With Congo red Principle Reagents Procedure and observation With methyl orange Principle Reagents Procedure and observation Normal values Clinical and diagnostic significance Design of laboratory work Practical 2 qualitative reaction of lactic acid presence in gastric juice Principle Reagents Material of investigation Procedure and observation Normal values Clinical and diagnostic significance Design of laboratory work Practical 3 Detection of blood and hemoglobin in gastric juice by diagnostic strips "hemophan" Principle Material for investigation Procedure and observation Normal values Clinical and diagnostic significance Design of laboratory work Practical 4 (in theory) tubeless method of acidity detection in gastric juice (test of acidity) Principle Reagents Material for investigation Procedure and observation Design of laboratory work theme 5.2. intracellular amino acid metabolism introduction the aim of the practical CLASS is SELF-STUDY QUESTIONS topics for reports Practical determination of aminotransferases activity in serum Principle Reagents Material of investigation Procedure and observation Calculation Normal values Clinical and diagnostic significance Design of laboratory work theme 5.3. ammonia metabolism and its disposal introduction the aim of THE practical CLASS is SELF-STUDY QUESTIONS topics for report Practical 1 determination of urea content in serum and urine Principle Reagents Material of investigation Procedure and observation Calculation Normal values Clinical and diagnostic significance Serum Urine Design of laboratory work Practical 2 determination of creatinine content in serum and urine Principle Reagents Material of investigation Procedure and observation Calculation Normal values Clinical and diagnostic significance Serum Urine Design of laboratory work theme 5.4. metabolism of some amino acids. their features and disorders (seminar) introduction the aim of THE practical CLASS is SELF-STUDY QUESTIONS topics for report tests case studies theme 6.1. STRUCTURE AND metabolism of purine and pyrimidine nucleotides introduction the aim oF the practical CLASS is SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical DETERMINATION OF URIC ACID CONCENTRATION IN SERUM AND URINE Principle Reagents Material of investigation Procedure and observation Calculation Normal values Clinical and diagnostic significance Serum Urine Design of laboratory work tests CASE STUDIES theme 7.1. nucleic acids synthesis and its regulation introduction the aim of THE practical CLASS is SELF-STUDY QUESTIONS topics for report Practical Analysis of chemical composition OF nucleoproteins Reagents Material for investigation Procedure and observation Design of laboratory work theme 7.2. protein biosynthesis and its regulation introduction the aim of THE practicalCLASS IS SELF-STUDY QUESTIONS topics for reports Practical determination of protein concentration in serum Biuret test Principle Material for investigation Reagents Procedure Calculation Normal values Clinical and diagnostic significance Hyperproteinemia Hypoproteinemia Design of laboratory work tests case studies theme 8.1. Structure and metabolism of carbohydrates. Glycogen metabolism Introduction SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical Detection of glucose in urine using "Glucophan" diagnostic strips Design of work: theme 8.2. Oxidation of glucose in aerobic conditions. Gluconeogenesis Introduction SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical 1 Principle Material of investigation Reagents Procedure Normal values Clinical and diagnostic significance Practical 2 DETECTION OF LACTIC ACID IN MUSCLE TISSUE BY UFFELMAN'S REACTION Principle The method is based on the complex compound interaction of violet iron phenolate with lactic acid to form yellow-green lactate of iron. Material for investigation Reagents Procedure Theme 8.3. Aerobic oxidation of glucose. Pentose phosphate pathway Introduction SELF-STUDY QUESTIONS TOPICS FOR REPORTS Practical Principle Material for investigation Reagents The glucose concentration determination Calculation Normal values Evaluation of the glycemic curve There are the following types of glycemic curves: Tests case studies theme 9.1. Structure and external metabolism of lipids Introduction SELF-STUDY QUESTIONS topics for reports Practical Principle Material for investigation Reagents Procedure Design of work: Theme 9.2. Intracellular metabolism of fatty acids and triacylglycerols Introduction the aim of the practical CLASS IS: SELF-STUDY QUESTIONS Topics for report Practical Principle Material for investigation Reagents Procedure Normal values Clinical-diagnostic significance Design of work: Theme 9.3. Intracellular metabolism of phospholipids and cholesterol. Transport of lipids in blood Introduction The aim of the practical CLASS is: SELF-STUDY QUESTIONS topics for report Practical Principle Material for investigation Reagents Procedure Normal values Design of practical: Tests case studies theme 10.1. The mechanisms of hormonal signal transduction. Classification of hormones. Hormones of pituitary gland. (seminar) introduction the aim of THE practical CLASS is: SELF-STUDY QUESTIONS topics for report theme 10.2. Hormones of hypothalamus, pituitary, thyroid, pancreas and parathyroid glands introduction the aim of the practical CLASS is: SELF-STUDY QUESTIONS topics for report Practical 1 Material for investigation Reagents Procedure Biuret test for Proteins Procedure Reaction for (-amino group detection Procedure Aromatic amino acids reaction The principle The principle Procedure Design of practical: Practical 2 Material for investigation Reagents Procedure Design of practical: Theme 10.3. Hormones of hypophysis, adrenal and sexual glands introduction the aim of the practical CLASS is: SELF-STUDY QUESTIONS topics for report Practical The principle Material for investigation: Reagents Procedure Design of practical: Tests case studies Introduction the aim of the practical CLASS is: SELF-STUDY QUESTIONS topics for report Practical 1 Thymol test The principle Reagents Material for investigation Procedure Normal values Clinical-diagnostic significance Design of practical Practical 2 (in theory) The principle Material for investigation Equipment Procedure (basic steps) theme 11.2. Iron exchange. Hemoproteins. The synthesis and breakdown of heme introduction the aim of the practical CLASS is: SELF-STUDY QUESTIONS topics for report Practical 1 DETERMINATION OF THE TOTAL BILIRUBIN AND ITS FRACTIONS CONCENTRATION IN BLOOD SERUM The principle Material for investigation Reagents Procedure Design of practical Practical 2 The principle Material for investigation Normal values Design of practical theme 11.3. Inorganic substances of the blood. Acid-base status introduction the aim of the practical CLASS is: SELF-STUDY QUESTIONS topics for report Practical 1 The principle Material for investigation Reagents Procedure Normal values Serum Urine Design of practical Practical 2 The principle The material for investigation Reagents Procedure Normal values Clinical-diagnostic significance Serum Urine Design of practical Tests case studies theme 12.1. Water-salt exchange. Normal and pathological components of urine introduction the aim of the practicalCLASS is: SELF-STUDY QUESTIONS topics for report Practical 1 Material for investigation Equipment Procedure The principle Procedure Normal value The influencing factors Design of practical: Practical 2 Material for investigation Equipment Procedure The principle Urine The principle Normal values Urine The principle Normal values Urine The principle Normal values Urine The principle Normal values Urine Normal values Urine Design of practical: Tests case studies Unit 1. "Structure, features and functions of proteins" Unit 2. "Structure, classification and role of vitamins" Unit 3."Enzymology" Unit 4. "Biological oxidation" Unit 5. "Amino acid and protein metabolism" Unit 6. "Structure and metabolism of purine and pyrimidine nucleotides" Unit 7. "Matrix biosynthesis" Unit 8. "Structure and metabolism of carbohydrates" Unit 9. "Structure and metabolism of lipids" Unit 10. "Hormonal regulation of human metabolism and functions" Unit 11. "Biochemistry of blood" Unit 12. "Biochemistry of kidneys" Unit 1. "Structure, features and functions of proteins" Unit 2. " Structure, classification and role of vitamins" Unit 3. "Enzymology" Unit 4. "Biological oxidation" Unit 5. "Amino acid and protein metabolism" Unit 6. "Structure and metabolism of purines and pyrimidines" Unit 7. "Matrix biosynthesis" Unit 8. "Structure and metabolism of carbohydrates" Unit 9. "Structure and metabolism of lipids" Unit 12. "Biochemistry of kidneys" The systematic name of oxidoreductases is formed as follows: Example 1 Example 2 Example 3 II Class. Transferase Group donor : group acceptor – transferable transferase group Example 1 Example 2 Example 3 III class. Hydrolase 1. Esterases – hydrolysis of ester bonds. 2. Lipases – hydrolysis of neutral fats. 3. Phosphatase – hydrolysis of monoesters of phosphoric acid. 4. Glycosidases – hydrolyses O- and N-glycosidic bonds. 5. Proteases, peptidases – hydrolysis of proteins and peptides. 6. Nuclease – hydrolysis of nucleic acids. The systematic name of hydrolases is formed: Hydrolyzable substrate : hydrolase release group Example 1 Example 2 Example 3 IV Class. lyase The systematic name of lyases is formed: Cleavable substrate : detachable group – lyase Example 1 Example 2 Example 3 Example 4 V CLASS. Isomerase Example 1 Example 2 Example 3 VI class. ligase The systematic name of the enzyme: Example 1 Example 2 Example 3 CHARACTERISTICS OFSOME BLOOD PROTEINS Fibrinogen Normal values Clinical and diagnostic significance (1-Globulins Acid (1-glycoprotein Normal values Clinical and diagnostic significance (1-Antitrypsin Normal values Clinical and diagnostic significance (1-Antichymotrypsin Normal values Clinicalanddiagnosticsignificance (2-Globulins С-reactive protein Normal values Clinicalanddiagnosticsignificance Haptoglobin Normal values Clinical and diagnostic significance (2-Macroglobulin Normal values Clinical and diagnostic significance Ceruloplasmin Normal values Clinicalanddiagnosticsignificance (-globulins Transferrin family Transferrin Normal values Clinical and diagnostic significance Lactoferrin Normal values Clinical and diagnostic significance Normal values of studied biocamical indicators Serum Urine Gastric juice Clearance of endogenous creatinine ContentS