Medical Biotechnology

General section Fundamentals of molecular medical diagnostics, pathological anatomy and nuclear physics. The concept of organ pathology. The methodologies for studying structural and functional gene variations. Main instruments used in molecular medical diagnostics, pathological anatomy and nuclear physics. Specific sections Course I (BIO/12 – Lucarelli) Potential and limits of the qualitative, quantitative and automated high throughput laboratory approach. Fundamentals of molecular medical diagnostics. Search and interpretation of DNA sequence variations. Use of molecular information for diagnostic, prognostic and therapeutic purposes. Concept of detection rate and mutational search of I, II, III and IV level. Nomenclature, classification and functional characterization of mutations. The epigenetic modifications of DNA. Structural, dynamic and functional aspects related to DNA methylation. DNA sequence variation search methodologies: ARMS, allelic discrimination, direct and reverse hybridization, PCR / OLA / SCS, minisequencing (SNaPshot). Automated DNA sequencing by cycle sequencing (Sanger method), pyrosequencing and next generation sequencing (NGS) approaches. Analysis of gene dosage: methodologies for investigating large deletions and duplications by MLPA. Automation in molecular and cellular diagnostics: liquid handlers, genetic analyzers, haematological analyzers and related software. Methods of DNA methylation analysis: HpaII / PCR, MS-MLPA, bisulfite / cloning / sequencing, RRBS, RLGS. Applications to molecular diagnosis of: pre-mRNA splicing diseases, cystic fibrosis, thalassemia, hemoglobinopathies, familial hypercholesterolemia, diseases of abnormal DNA methylation. Practical design and critical evaluation of molecular diagnostic tests. Course II (MED/04 – Giannini) Brief overview of genetic pathology. Review of the most relevant methodological achievements in molecular biology/molecular genetics. Methods to detect structural and numerical chromosomal aberrations. Methods to detect DNA rearrangements. Methods to detect point mutations. Set-up of pathology-related mutational screenings. Evolution of Sequencing technologies. Application of mutational screening approaches to different medical issues, with a particular focus on molecular oncology. High throughput molecular methods in molecular oncology. Course III (MED/08 – Corsi, Riminucci, Stoppacciaro) Prof. Stoppacciaro The histological diagnosis: tissue biopsy, tissue fixation, paraffin-embedding, frozen sections procedures, criteria of light microscopy examination of tissue sections. Techniques of molecular diagnoses at tissue level: FISH, in situ hybridization with immuno-enzymatic techniques; extraction of nucleic acids from fresh or paraffin-embedded tissues, laser-capture tissue micro-dissection, methods of DNA/RNA analysis, diagnostic relevance. Immunohistochemical staining: the principles of the technique, pitfalls and quality controls, diagnostic relevance. The diagnosis inflammatory response at the tissue level: morphological changes associated with acute or chronic inflammatory responses, diagnostic relevance of immunohistochemistry and flow cytometry, demonstration of infectious agents in tissues. Cancer diagnosis: the morphological diagnosis of cancer at tissue level, tumor invasiveness, histogenesis and classification of tumors, criteria for defining biological and clinical aggressiveness of cancer, concept and demonstration of minimal residual disease, targeted therapies for cancer, immune-oncology, liquid biopsy. Proff. Corsi, Riminucci Molecular pathology and molecular diagnosis: general principles and techniques. Molecular analysis of formalin-fixed, paraffin embedded tissues. Human Papilloma Virus infection and human pathology: cellular and molecular mechanisms, natural history, pathology and molecular diagnosis, with particular reference to the infection of the female genital tract. Molecular analysis in the diagnosis and prognosis of tumors of the haematopoietic and lymhoid tissues. Molecular analysis in the diagnosis and study of genetic skeletal disease, in particular Fibrous dysplasia of bone. Course IV (MED/36 – Signore) Instrumentation in nuclear medicine: main differences between nuclear medicine and radiology, collimators, linear scanners, gamma cameras and PET. Radiopharmaceuticals and radioisotopes: concept of radiopharmaceutical and notions of radiochemistry and radiopharmacology. Synthesis of the main iodine and technetium radiopharmaceuticals. In vitro quality controls of both chemical, biochemical and biological radiopharmaceuticals. Receptor imaging: examples of the preparation and use of radiopharmaceuticals for diagnostics and nuclear medical therapy; production and use of FDG. Receptoral imaging in infectivology, endocrinology, neurology, cardiology and oncology. Overview of radio-guided surgery. Overview of optical imaging and bioluminescence.

In order to understand properly the teaching content and to achieve the expected learning objectives, at the beginning of the didactic activity the student must possess the basic knowledge acquired during the frequency of previous courses in the field of cellular and molecular biology, biochemistry, embryology, anatomy, histology, general pathology and pathological anatomy.

Molecular Diagnostic W. B. Coleman, G. J. Tsongalis Humana Press Introduzione alla Medicina Molecolare Ross Dennis W. Springer Human Molecular Genetics Strachan T, Read . P. Garland Science Dai Geni ai Genomi Dale, Schantz, Plant EdiSES Robbins e Cotran. Le basi patologiche delle malattie. Edra Masson. Signore A, Paserio E, Chianelli M. Note di Medicina Nucleare per la pratica clinica. In: Trattato Italiano di Medicina Interna di P. Introzzi. M. Negri Ed; UTET Publ., Torino 1995, p. 1249-1292. Signore A, Mather SJ, Piaggio G, Malviya G, Dierckx RA. Molecular imaging of inflammation/infection: nuclear medicine and optical imaging agents and methods. Chem Rev. 2010 May 12;110(5):3112-45.

The teaching is based essentially on frontal lessons characterized by a strong interactive component between the teacher and the students. Other teaching methods may include working groups or individual activities focused on the analysis of methods and results of publications in international scientific journals. There are practical sessions and exercises.

The attendance is checked by the teacher by using updated lists provided by the Academic Office that will be signed by the students. The certificate of attendance of the teaching course is required for the student to be admitted to the final test.

The assessment consists of an oral exam with questions related to the course program and takes into account: a) knowledge of the topics related to the questions; b) adequacy of the answer to each question based on the expected skills at the end of the course; c) the logic of the answers; d) use of an appropriate language. The weight of each module is proportional to CFU.

Scientific articles related to topics included in the program will be recommended as a study tool for integrative purposes to frontal lessons and adopted texts.

The teaching is based essentially on frontal lessons characterized by a strong interactive component between the teacher and the students. Other teaching methods may include working groups or individual activities focused on the analysis of methods and results of publications in international scientific journals. There are practical sessions and exercises.