OVERVIEW

This integrated course should give the students theoretical-practical information to understand the fundamentals and the techniques at the basis of the biochemical analyses performed in the clinical laboratories. The different modules of this course analyse and integrate molecular and executive aspects concerning Human Systematic Biochemistry, Clinical Biochemistry, Radioimmunology and Clinical Chemical Techniques learning.

AIMS AND CONTENT

LEARNING OUTCOMES

The student should acquire basic and specialistic concepts about: • clinical information that can be obtained by measuring the alteration of selected  metabolic markers of glucidic, lipidic and nitrogen origin • relationship between protein structure/function characteristics and  possible utilization of proteins, including enzymes, as markers in clinical biochemistry analyses• assay of enzymes as tools to measure the concentration of some analytes •  mechanisms of hormone based signaling and relevant modification of validated parameters that can be measured • acid/base equilibria and changes indicative of altered tissue function. Clinical aspects of the  hepatic and kydney function parameters. Techniques utilized in the laboratory of clinical chemistry  including electrophoresis, spectrophotometry, potentiometry, nefelometry, radioimmunoassay.

AIMS AND LEARNING OUTCOMES

The general target of this course is to give the student the possibility to understand the links between the upgrade of the scientific discoveries on the   metabolism, the enzimology, the mechanism underlying the cell responses to local or systemic signals, the acid/base control and the choice of tissue function/lesion markers suitable to improve the diagnostics, prognostics and monitoring of therapeutic efficacy. To reach this target the teachers discuss the clinical meaning of the current parameters used to evaluate the functionality of the lungs, the liver, the muscle  and the kidney, as well as the problems connected to transfer new discoveries to clinical laboratory applications. The clinical biochemistry and the techniques of clinical chemistry issues are hence treated as dynamic and continuously evolving disciplines. Specific attention is given on the choice of innovative parameters, methodologies, techniques and devices utilized. Hence the students  will approach the world of work with a basic  understanding of the choice criteria to maintain some analytic tests, possibly sustained by a new assay, or  to change previously used tests with different systems endowed with a higher sensitivity or specificity, following their validation. Lastly we discuss critical issues and innovative future challenges in the clinical biochemistry field, including the expansion of the specific biomarkers and the increasing specificity to detect  tissue damage localized in different functional units of a single human organ. 

PREREQUISITES

The student should know the metabolic pathways occurring in humans and the mechanisms for their regulation. 

TEACHING METHODS

Teaching is performed by classroom-taught lessons with the aid of slides that are available to the students on  the  Aulaweb. For the part concerning the Clinical Chemistry techniques the student acquires information about the procedures utilized to obtain results that can be considered qualitatively and quantitatively acceptable, about the characteristics and the basis of the working principles of the devices utilized to carry out different clinical analyses.

SYLLABUS/CONTENT

Clinical Biochemistry: Clinical information obtained from specific alteration of electrolytic and acid/base equilibria. Markers of tissue function and damage: enzymes released by different cell sources. Glycemic profile: criteria to evaluate and manage the  mellitus diabetes; ematochemical and urinary parameters during diabetic ketoacidosis  and  hypoglicemic coma.Oral glucose tolerance test, glycated hemoglobin. Non protein nitrogen compounds of blood serum and information obtained from their concentration changes in the blood: urea, uric acido, creatinine, creatine, ammonia. Hepatic functionality:  biochemical alterations in different hepatopaties; bilirubine, transaminases. Kydney functionality: specific markers and tests, creatinine clearance, micro e macroalbuminuria. Clinical significance of altered urinary biochemical parameters .

Human Systematic Biochemistry: main metabolic pathways - mechanisms for the regulation of the metabolism of charboydrates, lipids and proteins.Metabolism integration and cell metabolic specialization. Liver: cellular organization  and functions-  specific metabolisms and their regulation (glycolisis and gluconeogenesis, glycogen; synthesis of different lipids, cholesterol, lipoproteins, aminoacids  and urea cycle) - relationship between the liver and other organs. Muscle: molecular bases of muscle contraction-specific metabolisms and their regulation-metabolic relationships with other tissues.  Adipose tissue: structure and function- specific imetabolisms and their regulation: signals for lipid mobilization-heat production. Intestinal metabolism: digestion of charboydrates, lipids, proteins,vitamins and nucleic acids.  Central nervous system: specific metabolisms and their regulation, syntesis of neurotransmitters. Blood: composition and role in the transport of metabolites- glycemia - transport of O2 e CO2 - ionic equilibrium,  plasma proteins and lipids  transport. Hormonal control: main classes of hormones and their functions- hormon  regulation of the  energetic metabolism- metaboliche changes following fasting and physical exercise.

Radioimmunology: Characteristics of a RIA test and  post-radioimmunologic era; overview of the structure/function of an antibody and production of  monoclonal and polyclonal antibodies; main isotopes used for RIA; calibration curve and interpolation of the results; kinetics of reaction and equilibrium in a  RIA reaction. Methods for the separation of free and bound fractions; elements of radiochemistry; definition of atom, nuclide, isotope and radioisotope. Radioactivity: types of radiations their energy and danger; radioactivity units; detection of beta radiation by  liquid scintillation and gamma radiation by solid scintillation; antigen labelling with tritium and 125  iodine; rules of behavior for persons working in a  RIA laboratory, knowing the  internal ed esternal contamination risks and use of  individual protection equipments; rules for the purchase, storage, transport  and disposal of radioactive material. Production of polyclonal antibodies; antibody titer and its importance in a  RIA method; antisera: specificity and affinity; production of a  RIAhybridoma:  polyclonal vs monoclonal antibodies;  IRMA (Immuno Radio Metric Assay or immunoradiometric method); Difference of IRMA vs RIA; Automation of data analysis; regression models: logit-log function, spline, polynomial with 4 parameters;  periodical monitoring of laboratory instruments: calibration and error limits. Outlines on the   control of quality in a  RIA laboratory.Types of errors in an analytical procedure  and their detection: rough error, random, systematic. Performance of an analytical system. Causes of data variability: common and special causes. outlines of the statistical control of quality: indexes of position, dispersion and shape; setting up of a control chart and characterization of the limits of alert and of action of the results obtained;  guidelines for a professional detecting errors or abnormal results during the assay. 

Techniques of Clinical Chemistry. Introduction to the Laboratory: the preanalytical phase, collection of the biological sample and criteria for its acceptability; types of samples; anticoagulants; treatment, delivery, centrifugation and storage. Equipments for analitical techniques: light carachteristics, interaction between materia and  radiant energy, Lambert/ Beer law, absorbance and titration curve; rudiments of photometry and spectrophotometry (with single or double beam spectrophotometers); turbidimetry; nephelometry. Clinical applications: spectrophotometry with direct and indirect methods; colorimetric reactions;  enzymatic reactions, primary and coupled reactions, enzymatic reactions  NAD e NADP dependent;  enzymatic-colorimetricreactions, Trinder end point method. Indirect Potentiometry: principles. Methods to determine the main parameters assayed in clinical chemistry (AST, ALT, gamma-GT, PCE and  dibucaine number, LDH,  total and direct Bilirubin, CPK, ALP, total and Pancreatic Amylase, Lipase, Urea and Ureic Nitrogen, Creatinine,  Uric Acid, calculation of endogenous clearance, Iron, Ferritin, Glucose, total Cholesterol, HDL e LDL, Triglycerides, APO A e B, Natrium, Kalium, Chloride, Calcium, Phosphorus, Magnesium): method and interpretation of a clinical datum, interferences. Quality of the analytical system: precision and preciseness of the analytical datum; specificity and sensitivity of a method; types of errors; calibrations, definition of limit of detection and sensitivity;  internal and external quality control; dilutions.

Electrophoresis and specifice proteins, biological materials  and their collections. General characteristics of proteins, overview of their structure and function, methods for their assay• automatic equipments and their functioning. Electrophoretic separation of  serum proteins•  Electrophoretic standard profile: protein fractions (albumin-α1-α2-β1-β2-γ globulins) •  specific proteins of the electrophoretic pattern.  Gammapathies, Electrophoretic Immunofixation  (EIF): principles and methods• Interpretation of the results, evaluation of normal and pathological patterns. Urinary electrophoresis. Search for Bence Jones protein • Methodology • Interpretatione of the results • Immunosubtraction and interpretation of the results. Definition of electrophoresis, electrophoretic techniques: principles, equipments and accessories (power supply,  electrophoretic chamber, buffer, dyes, densitometer) • Protein migration and conditioning factors (voltage,  frictional force,  migration rate, electrophoretic mobility; sample;  buffer solution; ionic strength;  Joule effect; electroendosmosis) • implementation methods: free or front phase; on sifting and non sifting support or zonal • zonal electrophoresis of serum proteins on paper,  cellulose acetate, agarose gel and in capillary • Methodology • Interpretation of migration and of graphics in capillary •evaluation of normal and pathological patterns• Comparison between methods.   

RECOMMENDED READING/BIBLIOGRAPHY

Biochimica clinica by Allan Gaw, Michael J. Murphy, Robert A. Cowan, M. Lobello, L. Rossi- Editore Elsevier

TEACHERS AND EXAM BOARD

Exam Board

MARIO PASSALACQUA (President)

ELISABETTA FRATERNALE

LAURA MARCOMINI

MADDALENA MASTROGIACOMO

ELENA ZOCCHI (President and Coordinator of Integrated Course)

LESSONS

Class schedule

CLINICAL BIOCHEMISTRY AND CLINICAL CHEMISTRY TECHNIQUES

EXAMS

EXAM DESCRIPTION

All the teachers involved in this class concomitantly participate to the oral exam and only the students that obtain a minimal examination pass mark  from  the whole committee obtain a positive evaluationa. The students have seven chances to pass the examination along each academic year.

ASSESSMENT METHODS

To assess the ability of the student to develop a specific topic proposed during the oral exam, it is highly appreciated his capacity to propose and discuss the links with other kindred disciplines, the deepening of crucial points by critical analysis of the information acquired during single parts of this integrated class, the use of technical terms to describe specific assays and the relevant instruments utilized to carry out different types of analyses, the correct framework of characteristics, potential and limitations  of alternative methods to measure clinical parameters.

Exam schedule