66154 - Biochemistry and Biotechnological Applications in Pathology

Course contents

Introduction

Revisiting the carbohydrate metabolism: glucose homeostasis, glucose transporters and main glycolytic enzymes, gluconeogenesis. Hormones and endogenous control (kinetic and thermodynamic) of glycolysis and gluconeogenesis in the liver. Regulation of glycolysis in cardiomyocytes and skeletal muscle cells. Central role of PDH in the regulation of energy metabolism. Tricarboxylic acid cycle: flux regulation, anaplerotic reactions and interaction with other metabolic pathways. Brief recall of the glycogen and pentose phosphate pathway. Lipid metabolism: central role of the acetyl-CoA carboxylase; nitrogen balance and amino acid metabolism; interaction with other pathways. An update on structure and function of mitochondria; the electrochemical gradient and its homeostasis; proton motive force and energy transduction. Structure and mechanism of aaction of the nanomachine, ATP synthase complex.

Mitochondrial diseases

A brief survey on the rare mitochondrial diseases; the pathogenesis of two or three mitochondrial diseases caused by mtDNA point mutations will be examined at the molecular level (NARP, Leigh's syndrome). Possible therapeutic approaches and biotechnology contribution.

Aging and neurodegenerative diseases

Aging theories with particolar emphasis to the radical and the mitochondrial ones. Reactive oxygen and nitrogen species and damages to the biological structures. Hypoxia and damage to cells and tissues. Cells and animal models of senescence.

Metabolic changes associated with the Alzheimer disease; cardiovascular dysfunction and beta amyloid hypotheses. Insight into the dysfunction of cells and organules exposed to beta amyloid peptides. PET and magnetic resonance as non invasive tools for AD diagnosis.

Diabetes and insulin

Type I and II diabetes. Metabolic changes, glycemia, chetosis. Diet and insulin treatment as therapeutic means. Structure and functional properties of insulin. Insulin extraction from pig pancreas, purification and humanization; preparation of insulin by chemical synthesis. Preparation of the hormone by DNA recombinant techniques and site directed mutagenesis to obtain slow and fast insulins. Technical and economical analysis of the different methods of insulin preparation to be used therapeutically.

Cardiac metabolism and ischemia-reperfusion of the heart

Energy substrates of cardiomyocytes in physiology and in hypoxic conditions. Hyperlacticemia and ph changes in the ischemic heart; transient metabolic and mitochondrial adaptation to ischemia; oxidative metabolism in the early reperfusion of the ischemic heart. Brief analysis of drugs used to treat myocardial infarction. Ischemic preconditioning.

Most frequent metabolic hallmarks in tumor cells

Analysis of the carbohydrate metabolism (Warburg effect). Metabolism under hypoxia and the transcription factor HIF. Brief presentation of the protein p53 and consequences of her mutations.

Three practicals in the laboratory to evaluate enzyme kinetic parameters, rates of cells respiration, and measure of the mitochondrial membrane potential.

Readings/Bibliography

Prints of the ppt slides projected during the lectures will be supplied to the learners, and references to books, reviews and original papers from which tables and figures are taken will be provided.
Indicative reading for learners:
Textbooks: The biochemical principles of Lehninger, 6° Ed. Nelson DL e Cox MM (Zanichelli)
Biochemistry, 5° Ed. Devlin T.M. (Edises)
Biochemistry, 7° Ed. Berg JM, Timoczko JL, Stryer L. (Zanichelli)
Medical Biochemistry Lieberman M., Marks AD (Ambrosiana)
Biochemistry 4° Ed Voet & Voet (Wiley)

Teaching tools

Most time will be spent in the lecture theater to comment ppt slides and movies. Sometimes students will collaborate with the teacher by writing reactions and schemes on the black-board.

Three practical experience in the laboratory of Biochemistry (DIBINEM) will be planned.

Office hours

See the website of Giancarlo Solaini