Ateneo - UPO

The course addresses biochemistry applied to pathophysiology and laboratory medicine, integrating the biological significance of biomarkers with the methods used for their measurement and interpretation.The course is structured around the relationship between pathophysiological process, biomarker, biological sample and analytical technology. Protein, enzyme, metabolic, hormonal and genomic biomarkers will be examined, together with the principles of the main methods used for their determination.Selected diseases will be used as models to understand how biochemical and molecular mechanisms guide the choice of biological material and diagnostic method, as well as the interpretation of laboratory results. Particular attention will be devoted to inherited metabolic disorders, molecular diagnostics in oncology, hormonal regulation and the biochemical alterations associated with metabolic syndrome

M. Ciaccio (ed.), Trattato di Biochimica Clinica e Medicina di Laboratorio, EdiSES, 1st edition, 2021, 894 pp.Lecture slides and supplementary materials provided by the lecturer are also part of the reference material for exam preparation.

The course aims to provide students with the biochemical and methodological knowledge required to understand the role of biomarkers in pathophysiology and laboratory medicine.

In particular, the course aims to enable students to:
- understand the biological and diagnostic significance of the main protein, enzyme, metabolic, hormonal and genomic biomarkers;
- relate alterations in laboratory parameters to the biochemical and molecular processes underlying major pathophysiological conditions;
- understand the criteria guiding the choice of the most appropriate biological sample and analytical method according to the biomarker and the diagnostic question;
- identify the main sources of variability and the limitations of laboratory investigations;
- understand the process of biomarker development and validation, from experimental research to clinical application;
- acquire the tools required to interpret simple biochemical, clinical and molecular results in an integrated manner.

The course integrates basic knowledge, laboratory medicine applications and elements of translational biomedical research, showing how the understanding of biochemical mechanisms can guide the development and use of diagnostic tools

Basic knowledge of biochemistry, cell biology and molecular biology is required to adequately follow the course.
In particular, students should be familiar with:
- the structure and function of the main classes of biomolecules;
- the major metabolic pathways and their regulatory mechanisms;
- the principles of enzyme catalysis;
- signal transduction mechanisms;
- DNA replication, transcription and translation, as well as the main mechanisms regulating gene expression;
- cell organization and the functions of the main cellular organelles.
This background is necessary to understand the biochemical and molecular basis of pathophysiological alterations, the significance of biomarkers and the principles of the diagnostic methods covered in the course.

The course consists of lectures supported by slides and additional teaching materials.
During the lectures, questions, quizzes, application exercises and guided discussions of representative cases will be proposed to connect biochemical and molecular mechanisms with the choice of the most appropriate biomarker, biological sample and diagnostic method.
Interactive activities are intended to encourage active participation, critical thinking and the ability to apply acquired knowledge to simple problems in clinical biochemistry and molecular diagnostics.
Lecture slides, supplementary materials and any self-assessment activities will be made available on the Moodle DIR platform

Lecture slides, supplementary materials, notices and information concerning the course and examination procedures will be made available on the Moodle DIR platform.The examination always refers to the syllabus of the current academic year, including for students enrolled in previous academic years.Students with disabilities, Specific Learning Disorders (SLD) or Special Educational Needs (SEN) may request the dedicated services and support measures provided by the University by contacting the relevant University Office and consulting the dedicated section of the UPO website. After contacting the University Office, students may contact the lecturer to agree on the application of the appropriate measures to teaching activities and examination procedures

The examination consists of an individual, in-person written test administered through the Moodle DIR platform in a computer room.
The test lasts 45 minutes and consists of 35 questions covering all the course topics. The questions may include:
- multiple-choice questions;
- fill-in-the-blank questions;
- matching, ordering or classification tasks;
- short application-based questions aimed at assessing the ability to relate a pathophysiological process to the appropriate biomarker, biological sample and diagnostic method.

The examination assesses:
- knowledge and understanding of the course contents;
- the ability to relate biochemical and molecular mechanisms to biomarker alterations;
- the ability to apply acquired knowledge to simple problems in clinical biochemistry and molecular diagnostics;
- the ability to identify the potential, limitations and main sources of variability of laboratory investigations.

Incorrect and unanswered questions do not incur penalties. The overall score is converted into a grade out of thirty, and the examination is passed with a grade of at least 18/30.

A passing grade requires knowledge of the fundamental concepts and the ability to correctly establish the essential relationships between pathophysiological process, biomarker, biological sample and diagnostic method. Higher grades require comprehensive and in-depth knowledge, the ability to integrate different topics and the autonomous application of knowledge to more complex questions. Honours may be awarded for an excellent performance.

The course addresses biochemistry applied to pathophysiology and laboratory medicine. Its central theme is the relationship between pathophysiological process, biomarker, biological sample and analytical technology. Selected diseases will be used as models to understand how biochemical and molecular mechanisms guide the choice of biological material, diagnostic method and interpretation of results.1. Biomarkers and laboratory investigationsBiological and diagnostic significance of protein, metabolic and genomic biomarkers. Diagnostic, predictive and prognostic biomarkers, and biomarkers of treatment response and disease monitoring. Biomarker development and validation: candidate identification, preclinical and clinical validation, standardisation and implementation in clinical practice. Potential and limitations of laboratory investigations.The biological sample as the starting point of laboratory investigation: whole blood, serum, plasma, urine, cerebrospinal fluid, dried blood spots and tissue biopsies. Criteria guiding sample selection according to the biomarker, diagnostic question and analytical technology; main pre-analytical factors affecting the result.2. Protein and enzyme biomarkersMain plasma proteins and their diagnostic significance. Inflammatory biomarkers: C-reactive protein, procalcitonin and fibrinogen; sample type, kinetics and contexts of use. Tumour markers: criteria for use, applications and diagnostic limitations. Proteomics: general principles and translation from biomedical research to clinical diagnostics.Criteria for using an enzyme as a biomarker: intracellular localisation, tissue distribution and isoenzymes. Spectrophotometric and kinetic principles for measuring enzyme activity. Physiological and pathological factors affecting blood enzyme levels. Organ-related enzyme patterns: transaminases, creatine kinase, lactate dehydrogenase, alkaline phosphatase, gamma-glutamyl transferase, amylase, lipase, trypsin and cholinesterase. Diagnostic significance of isoenzymes. Biomarkers of myocardial injury: myoglobin and troponins. Acute myocardial infarction as a model integrating serial sampling, biomarker kinetics, immunoassays and point-of-care devices.3. Metabolic biomarkers and inherited metabolic disordersDefinition of metabolome and metabolomics; targeted and untargeted approaches. Main analytical platforms: nuclear magnetic resonance spectroscopy, mass spectrometry and liquid chromatography. Samples, potential and limitations of the different technologies. Applications in diagnosis, patient stratification and the study of ageing.Biochemical and pathophysiological classification of inherited metabolic disorders: accumulation or intoxication disorders, defects in energy production or utilisation, and disorders of complex molecule metabolism. Principles of newborn screening, criteria for disease selection, collection using dried blood spots and the role of tandem mass spectrometry. Integrated diagnostic approach: metabolic biomarker, enzyme or protein assay and genetic-molecular confirmation.Disease models: phenylketonuria, as an accumulation aminoacidopathy; Maple Syrup Urine Disease, as a defect in branched-chain amino acid catabolism; MCAD deficiency, as a beta-oxidation defect; Gaucher disease, as a lysosomal storage disorder; congenital adrenal hyperplasia, as a disorder of steroidogenesis. For each model, the biochemical mechanism, biomarkers, sample and diagnostic technologies will be considered, without a systematic clinical description.4. Genomic biomarkers and molecular diagnosticsClassification and functional consequences of the main genetic alterations investigated in molecular diagnostics. Potential and limitations of molecular tests. Selection of biological material according to the type and purpose of the investigation: germline DNA, tumour DNA, circulating cell-free DNA and RNA. Sensitivity and specificity of molecular tests. Principles of the main techniques used in molecular diagnostics. Molecular biomarkers in colorectal and lung cancer as models integrating molecular alteration, biological sample, tissue biopsy, liquid biopsy and diagnostic technology.5. Biochemistry of hormonal regulationHormones as biomarkers: sample selection, pre-analytical stability, interferences and timing of collection in relation to secretion rhythms.Vasopressin and oxytocin: structure, mechanism of action and metabolic effects. Renin-angiotensin-aldosterone system and regulation of water-salt balance and blood pressure.Hypothalamic-pituitary axes and peripheral targets: CRH-ACTH-adrenal hormone axis, glucocorticoids and mineralocorticoids; GnRH-LH/FSH-sex hormone axis, gonadotropins, progesterone, oestrogens and androgens; TRH-TSH-thyroid hormone axis, regulation, synthesis, mechanism of action, metabolic effects, hypothyroidism and hyperthyroidism; GHRH-GH-IGF-1 axis, metabolic effects and major alterations. Pro-opiomelanocortin-derived peptides: melanocortins, endorphins and lipotropins.Regulation of calcium and phosphate metabolism: parathyroid hormone, calcitriol and vitamin D; biosynthesis, receptors, signal transduction and regulation of calcium homeostasis.Neurohormonal regulation: signalling by the sympathetic and parasympathetic nervous systems; catecholamines and the stress response; dopamine and serotonin; overview of the main gastrointestinal hormones.6. Regulation of energy metabolism and metabolic syndromeRegulation of catabolic and anabolic processes. PI3K-AKT-mTOR and AMPK pathways. Sirtuins. Insulin signalling and systemic metabolic effects. Transcriptional control of energy metabolism. Alterations in insulin signalling and glucose and lipid homeostasis. Biochemical and molecular basis of metabolic syndrome. Overview of caloric restriction, energy metabolism and ageing.Integration of the gender dimensionWhere relevant, differences related to biological sex and gender determinants will be considered in endocrine and metabolic regulation, biomarker variability and the interpretation of laboratory results.

Knowledge and understanding
At the end of the course, students will be able to:
- describe the biological and diagnostic significance of the main classes of protein, enzyme, metabolic, hormonal and genomic biomarkers;
- understand the biochemical and molecular basis of alterations in laboratory parameters in the physiological and pathological conditions covered by the course;
- know the characteristics of the main biological samples and the criteria guiding their selection;
- understand the principles of the main methods used in clinical biochemistry, metabolomics and molecular diagnostics;
- describe the main mechanisms regulating hormonal and energy metabolism;
- identify the main sources of pre-analytical and analytical variability and the limitations of laboratory investigations.

Applying knowledge and understanding
Students will be able to:
- relate a biochemical or pathophysiological process to the most appropriate biomarkers;
- identify the most suitable biological sample and analytical method according to the biomarker and the purpose of the investigation;
- interpret simple biochemical, metabolic, hormonal and molecular profiles;
- integrate information concerning biochemical mechanism, biomarker, biological sample, technology and diagnostic timing;
- apply acquired knowledge to simple questions and problems in clinical biochemistry and molecular diagnostics;
- recognise pre-analytical and analytical conditions that may compromise the quality and informational value of a result.

Making judgements
Students will be able to:
- critically assess the appropriateness of a biomarker, biological sample and diagnostic method in relation to a specific question;
- compare the potential and limitations of different analytical approaches;
- distinguish the association between a biomarker and a pathological condition from its actual diagnostic, prognostic or predictive value;
- identify possible sources of error or incorrect interpretation of laboratory results.

Communication skills
Students will be able to use the terminology of clinical biochemistry and molecular diagnostics appropriately and to clearly describe the biological and diagnostic significance of a biomarker, the principle of a method and the rationale underlying sample selection.

Learning skills
Students will be able to:
- use the teaching materials, reference textbook and scientific sources provided to independently consolidate and expand their knowledge;
- integrate knowledge from biochemistry, molecular biology, physiology and laboratory medicine;
- update their knowledge in response to developments in biomarkers and diagnostic technologies.

The minimum passing level requires knowledge of the fundamental concepts, the correct association between pathophysiological process, biomarker, biological sample and analytical method, and the ability to apply this knowledge to simple situations.
An advanced level requires comprehensive and in-depth knowledge, the ability to independently integrate different topics, critically compare diagnostic approaches and apply acquired knowledge to more complex questions.