GENERAL AND APPLIED BIOCHEMISTRY
Academic year and teacher
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- Versione italiana
- Academic year
- 2017/2018
- Teacher
- RICCARDO GAVIOLI
- Credits
- 12
- Didactic period
- Primo Semestre
- SSD
- BIO/10
Training objectives
- Biochemistry is a branch of life science which deals with the study of chemical reactions and their organization that occur in living cells and organisms. It takes into account the studies related to the nature of the chemical constituents of living matter, their transformations in biological systems and the energy changes associated with these transformations. Students will be able to apply their knowledges for the evaluation of the impact of structural variations of biomolecules on their function, the role of modulators on biomolecules in the field of therapeutics and will allow the understanding on the impact of the variations of physiological conditions on metabolic pathways.
The objective of the “Biochemical and clinical analysis” part is analize the principal biochemical technologies and the modern diagnostic methodologies, with reports about available techniques for the bio-molecules study. The course is based on the examination of different biochemical methodologies (chromatography, electrophoresis, spectroscopy, etc) with particular attention to real and clinical examples and will constitute the bases for possible applications. Prerequisites
- Basic knowledge of Biology, general and Organic Chemistry.
To take the final examination (learning assessment procedures), students must have passed the exam of Organic chemistry. Course programme
- The course is divided in two parts of 6 CFU each, corresponding to 48 hours of lectures per part.
Part 1. Biochemistry
Topics of the lessons are:
Introduction to Biochemistry (2 hours): Cell and organelles, composition of living matter, configuration of the carbon atom, molecules, covalent and non-covalent bonds, properties of water.
Nucleotides (2 hours): structure and function
Proteins: Structures, functions and catalysis (10 hours).
Amino acids. Proteins. Structure and function of hemoglobin and myoglobin. Structure and function of immunoglobulins. MHC and TCR. Enzymes. Allosteric enzymes and enzymes regulated by covalent modifications.
Biological Membranes and Transport (2 hours): The constituents of plasma membranes. Structure and function of membranes. The membrane lipids. Fluidity. Permeability. Passive and active transport:
Membrane receptors (4 hours): function, specificity and signal transduction: receptors with intrinsic kinase activity (insulin), cAMP, diacylglycerol and inositol triphosphate.
Bioenergetics and Metabolism (26 hours):
Basic concepts of metabolism: Catabolism and anabolism. The transfer of phosphate groups. Structure and function of ATP. The electron carriers (NADH, NADPH and FADH2). Coenzyme A.
Degradation and synthesis of glucose. Synthesis of pentose phosphate and NADPH.
Glycogenolysis and glycogen synthesis. Regulation of sugar metabolis. Hormonal regulation of sugar metabolism. Citric acid cycle. Functions, energy balance and regulation. Degradation of lipids. The ketone bodies. Degradation of amino acids and proteins. Protein diet and cellular proteins. Ethanol metabolism. Oxidative phosphorylation. Biosynthesis of lipids.
Integration of Metabolism (2 hours): Metabolic Inter-relations between different organs. Fasting-feeding cycle. Hormonal regulation.
Part 2. Biochemical and clinical analysis
Topics of the lessons are:
Introduction (2 hours): Lab safety: safety for patients, visitors and personnel. Analytical variability.
Emocromo cytometric analysis (4 hours): Hematopoiesis and blood cells. Cell counts (multiparameter counters), Hct value, Hb evaluation, MCV value (flow cytometry), blood film.
Electrophoresis (10 hours): General principles. Agarose gel, cellogel, polyacrylamide gel, 2D gel, capillary electrophoresis. Examples: Hb analysis, serum proteins dosage, new markers, DNA analysis.
Chromatography (10 hours): General principles. Classic column chromatography (normal and invers phase). Adsorption, dimensional exclusion, ionic exchange and affinity chromatography. HPLC (High Performance Liquid Chromatography): pumps, injectors, columns, detectors. Clinical applications: nucleic acid and protein separation, analysis of plasma-proteins, monitoring of normal and pathological hemoglobins.
Spettroscopy (2 hours): Spectrophotometry: absorbance, Lambert-Beer equation. Applications: nucleic acid and protein concentration, Hb blood concentration, use of spectrophotometer in ELISA and in enzyme-colorimetric reactions.
Mass spectrometry (4 hours): Function and characteristics of mass spectrometers. Ion source (MALDI and ESI), analyzers (quadrupole, ion trap, TOF), detectors. Applications: differential expression and identification of interesting proteins (proteome study; biomarker's research).
Immunochemistry (6 hours): Antibodies and antigens (structure and function). Radio Immuno Assay (RIA), Enzyme Linked Immuno Sorbent Assay (ELISA). Clinical applications: tumor markers, cardio-vascular markers, diagnosis of diabetes mellitus type 1 and celiac disease, fast assays (pregnancy test).
Molecular biology techniques (10 hours): Polymerase Chain Reaction (PCR), DNA sequencing. Clinical applications: diagnosis of genetic diseases (Thalassemia) and non invasive pre-natal diagnosis. Didactic methods
- The course is organized in frontal lessons on all the course’s topics.
Learning assessment procedures
- Oral examination. The aim of the oral examination is to test the level of knowledge and deepening of the topics of the course program and the reasoning skills developed by the student on issues related to biochemistry and applied biochemistry. The student can choose to do partial tests of the two modules that will be valid for the final evaluation exam test. The evaluation is expressed in thirtieths (minimum grade 18)
Reference texts
- Biochemistry:
Scheme and Figures provided by the docent.
One of the following books:
"Biochimica" di Berg-Tynoczko-Stryer, Zanichelli
"I principi di Biochimica di Lehninger" di Nelson-Cox, Zanichelli
"Principi di Biochimica" di Garret&Grisham, Piccin
"Fondamenti di Biochimica" di Voet-Voet-Pratt, Zanichelli
“Biochimica con aspetti clinico-farmaceutici” di Devlin, Edises
Biochemical and clinical analysis:
Scheme and Figures provided by the docent.
One of the following books:
“Biochimica applicata” di Stoppini e Bellotti, EdiSES
“Metodologie biochimiche” di Bonaccorsi di Patti, Casa Editrice Ambrosiana
“Medicina di laboratorio” di Federici, McGrawHill
“Compendio di scienze del laboratorio clinico” di Hubbard, Verduci Editore
