APPLIED PATHOPHYSIOLOGY

Degree course: 
Corso di Second cycle degree in Biotechnology for the Bio-based and Health Industry
Academic year when starting the degree: 
2022/2023
Year: 
2
Academic year in which the course will be held: 
2023/2024
Language: 
English
Credits: 
6
Period: 
First Semester
Standard lectures hours: 
52
Requirements: 

Students are required to have previous knowledge on cell, tissue and organ physiology, biochemistry, physics, and organic chemistry, molecular biology. Concerning the section of Pathology, a strong background in immunology is required. By the end of the lessons, students will be able to use basic concepts of Molecular Physiology and Molecular Pathology in the context of biotechnologies.

Final Examination: 
Orale

Oral examination with evaluation in thirty. The exam is individual for the physiology and the pathology program.
During the Physiology course, each student is required to find a review/scientific article based on different topics/techniques covered during the lesson and write a two-page report (three reports during the entire course) based on it. The Physiology exam consists of an oral Journal Club presentation based on an original research paper on a neurodegenerative disease involving a receptor or channel described during the course. About 70% of the evaluation is based on the knowledge concerning the topics included in the program. The remaining part is based on the ability of the student to clearly answer the questions, use a proper terminology, and on the work done on paper presentation.
For the pathology exam, the student must sustain an oral exam that concerns questions related to the theoretical and technical aspects discussed during the lessons. The final evaluation will consider the knowledge acquired and understanding (40%), the application of the knowledge acquired, by discussing the specific topic. And providing related examples/applications (40%), communication skills (20%). Precision in terminology and content correctness are required to pass the exam. Ability to integrate the different knowledge acquired and contents will be considered to get higher marks (29-30 and honors).

Assessment: 
Voto Finale

Specific knowledge: Improve the knowledge on basic homeostatic and pathologic processes by taking advantage of the progress on Molecular Physiology and Molecular Pathology research. The purpose is to provide the students updated information on the function of molecules, cells and tissues, which is the prerequisite for understanding the mechanisms involved in maintaining homeostasis, and whose alterations, both at cellular and molecular levels, contributes to the development of diseases.
For the Physiology section, particular attention will be dedicated to the nervous system, in particular to the behavior of the cell membrane and its protein, the role of specific ions in cell function, cell transport systems, cell communication, receptors and signaling. Specific cell types (i.e. neuron and glia cells) are used as examples to describe complex molecular processes which are the base of cell homeostasis. The course will provide a comprehensive presentation of the various cellular and electrophysiological methods used to study cellular communication between neurons.
For the pathology section, particular attention will be dedicated to the molecular and cellular process in cancer and tumor immunology. To this end the contribution of soluble and structural components, tumor cells, stromal cells, and immune cells, as mediators of cancer onset and progression, will be dissected.

Contents section of Physiology

Cell physiology-Cell homeostasis: Sodium, Calcium, Chloride, Volume, Osmolarity, Excitation -Inhibition balance

Cell membrane physiology:

Neuron-Glia:
-Principles of cell communications
-Basic electrical properties
-Neurotransmitter

Receptors
-Metabotropic
-Ionotropic

Ion channels
-Chloride channels, Sodium channels, Potassium channels.
-Ligand activated channels,
-Voltage activated channels,
-Calcium activated channels,
-cAMP activated channels,
-TRP Channels

Calcium and the cell:
-Calcium-binding proteins
-Calcium pumps and exchangers
-Calcium receptors
-Calcium transients
-Stimulus-secretion coupling

Solute Carriers and neurotransmitter transporters

Molecular events at the synapse

Techniques for functional investigations of membrane protein and the synaptic events such as Two-electrodes voltage clamp, Patch clamp, Multi Electrode Array, in vivo electrophysiology, optogenetics, voltage clamp fluorimetry.
Laboratory activities will be aimed at presenting the main techniques covered in the course.

Contents section of Pathology

Overview on general pathology and immunology: cell adaptation to damages, acute and chronic inflammation, tumor biology. Introduction to applied pathophysiology.

Molecular pathways in oncology. Overview of the most relevant signaling pathways involved in tumor onset and progression from a tumor intrinsic and tumor extrinsic point view.

Cancer Associated Fibroblasts (CAFs): classification, differences with normal fibroblasts (NFs). CAF phenotypes. CAFs in tumor progression, immunosuppression, angiogenesis, and tumor metabolism. Experimental procedures to study CAFs. Therapies targeting CAFs.

Cancer Stem Cells: classification, relevance in tumor biology and anti-tumor therapy. Circulating tumor cells. CSCs and metastasis. Experimental procedures to study CSCs. Therapies targeting CSCs.

Tumor spheroids and organoids in pathophysiology: generation of tumor spheroids and related biomedical applications; generation of tumor organoids and biomedical applications.

Animal models in oncology: murine models of chemically induced carcinogenesis, orthotopic cancer murine models, transgenic mouse models (selected examples), humanized mouse models.

Cancer immunotherapy. General concepts for active and passive immunization. Immune checkpoints. Strategy in cancer immunotherapy: bacterial products, immune-cytokines, vaccines, adoptive cell transfer with T or NK cells. Chimeric antigen receptors: CAR-T and CAR-NK cells. Combining immunotherapy with chemotherapy and anti-angiogenesis. Side effect of immunotherapy: Cytokine Release Syndrome (CRS).

Extracellular vesicles (EVs): definition, classification, biological activities, experimental procedures to study EVs.

Advanced flow cytometry and cell sorting. Basic of flow cytometry: fluorescence, monoclonal antibody, antigen density, determination of the stain index. Components of a flow cytometer and cell sorter: flow chamber, optics and filters, signals. Flow panel design and compensation. Specific application and examples of flow cytometry: apoptosis, cell cycle, oxidative stress, intracellular and intranuclear factors, immunophenotyping.

Applications: Immunophenotyping by multicolour flow cytometry: panel design, isolation of PBMCs from whole blood, antibody staining, FACS acquisition and flow data analysis.

Contents section of Physiology

Cell physiology-Cell homeostasis: Sodium, Calcium, Chloride, Volume, Osmolarity, Excitation -Inhibition balance

Cell membrane physiology:

Neuron-Glia:
-Principles of cell communications
-Basic electrical properties
-Neurotransmitter

Receptors
-Metabotropic
-Ionotropic

Ion channels
-Chloride channels, Sodium channels, Potassium channels.
-Ligand activated channels,
-Voltage activated channels,
-Calcium activated channels,
-cAMP activated channels,
-TRP Channels

Calcium and the cell:
-Calcium-binding proteins
-Calcium pumps and exchangers
-Calcium receptors
-Calcium transients
-Stimulus-secretion coupling

Solute Carriers and neurotransmitter transporters

Molecular events at the synapse

Techniques for functional investigations of membrane protein and the synaptic events such as Two-electrodes voltage clamp, Patch clamp, Multi Electrode Array, in vivo electrophysiology, optogenetics, voltage clamp fluorimetry.
Laboratory activities will be aimed at presenting the main techniques covered in the course.

Contents section of Pathology

Overview on general pathology and immunology: cell adaptation to damages, acute and chronic inflammation, tumor biology. Introduction to applied pathophysiology.

Molecular pathways in oncology. Overview of the most relevant signaling pathways involved in tumor onset and progression from a tumor intrinsic and tumor extrinsic point view.

Cancer Associated Fibroblasts (CAFs): classification, differences with normal fibroblasts (NFs). CAF phenotypes. CAFs in tumor progression, immunosuppression, angiogenesis, and tumor metabolism. Experimental procedures to study CAFs. Therapies targeting CAFs.

Cancer Stem Cells: classification, relevance in tumor biology and anti-tumor therapy. Circulating tumor cells. CSCs and metastasis. Experimental procedures to study CSCs. Therapies targeting CSCs.

Tumor spheroids and organoids in pathophysiology: generation of tumor spheroids and related biomedical applications; generation of tumor organoids and biomedical applications.

Animal models in oncology: murine models of chemically induced carcinogenesis, orthotopic cancer murine models, transgenic mouse models (selected examples), humanized mouse models.

Cancer immunotherapy. General concepts for active and passive immunization. Immune checkpoints. Strategy in cancer immunotherapy: bacterial products, immune-cytokines, vaccines, adoptive cell transfer with T or NK cells. Chimeric antigen receptors: CAR-T and CAR-NK cells. Combining immunotherapy with chemotherapy and anti-angiogenesis. Side effect of immunotherapy: Cytokine Release Syndrome (CRS).

Extracellular vesicles (EVs): definition, classification, biological activities, experimental procedures to study EVs.

Advanced flow cytometry and cell sorting. Basic of flow cytometry: fluorescence, monoclonal antibody, antigen density, determination of the stain index. Components of a flow cytometer and cell sorter: flow chamber, optics and filters, signals. Flow panel design and compensation. Specific application and examples of flow cytometry: apoptosis, cell cycle, oxidative stress, intracellular and intranuclear factors, immunophenotyping.

Applications: Immunophenotyping by multicolour flow cytometry: panel design, isolation of PBMCs from whole blood, antibody staining, FACS acquisition and flow data analysis.

Convenzionale

Front lessons with ppt files and slides projection, seminars, and videos. Throughout the teaching timing, appropriate scientific papers are provided to the students.

Professors