Face to Face:

Teaching of theory consists of lectures in and laboratory practical. 

Laboratory exercises (in 2  student groups of students,2 instructors per group of 10 students) composing the students’ practical are complementary to the lectures and they aim to familiarize the student with the operation of simple laboratory instruments and the experimental procedures that are often used in diagnosis as well as to help the students comprehend concepts that are not easily presented theoretically (learning based on practical experience).

Attendance of lectures is not mandatory.  Attendance of Laboratory Practical Exercises is obligatory.

Information and Communication Technologies are used for the preparation of the lecture material, the online information and provision of supplementary learning material to students.

Specifically:

• Common software (e.g. MS powerpoint) is used to prepare lecture material and display slides and videos.
• The study guide (detailed supplementary material & additional bibliography), the theory and protocols of the laboratory exercises, the slides of each lecture as well as relevant videos and scientific articles made available electronically and online to students through the e-class system of our university.
• Information about the course, instructors, and their research interests and in general the Laboratory of Biology of the Faculty of Medicine are available online through the e-class system of our university.
• Common software (e.g. MS excel) is used to statistically process student assessment.

Announcements, information etc are available online via e-class. Communication is also done via e-mail and MS-Teams.

The language of assessment is Greek.

Evaluation methods.

A. For the laboratory practical: Laboratory Assignment Reports, Written Examination at the end of the semester with multiple choice questions and problem solving.

The participation of students in the laboratory exercises as well as the written report of the results of the exercises is mandatory. The report includes the results (presented in tables and diagrams, and the conclusions (e.g. if the results were expected, if not why, sources of possible errors in the experiments) as requested by each exercise. At the end of each exercise, the written report is checked by the instructors. At the end of the semester the students are examined in the content of the Laboratory practical exercises. The examined material consists of the theory, the methodology and the ways results are processed as included in the Guide of the Laboratory Practical or presented by the instructors during the exercises. Only the students that have successfully completed the laboratory exercises can participate in the written laboratory examination. Success in the laboratory examination is a prerequisite for participation in the course exams.

B. For the lecture material: The course exams are written, lasting 2 hours, and consist of critical or short answer questions as well as multiple choice questions The material to be examined is lectures and tutorial material as described above. Only those students who have successfully passed the Laboratory exams have the right to participate in the course exams.

Final Grade:

The final grade of the course is calculated as the sum of 80% of the grade of the written course exams and 20% of the grade of the Laboratory Exercises written exams.

The course aims to highlight the role of molecular biology in modern medicine. It focuses on the investigation of the nature, structure and properties of genetic material, on mutagenic factors (endogenous and exogenous) responsible for DNA damage, on the mechanisms of damage repair and on diseases that arise as a result of dysfunction of these mechanisms. Particular emphasis is placed on the analysis of the flow of genetic information and the mechanisms of regulation of gene expression in prokaryotic and mainly eukaryotic organisms. The properties of chromatin (euchromatin and heterochromatin), chromosomes, the genome and the mechanisms by which genetic variation arises, as well as the evolution of genes and genomes, are analysed. In addition, epigenetic modifications of the genome (DNA methylation, histone modifications, short and long non-coding RNA molecules) and their involvement in homeostasis and in diseases and malignancies are analysed and new data for their use in therapeutic interventions are provided. Particular emphasis is placed on new technologies for genome analysis and their role in the diagnosis, prevention and treatment of diseases and neoplasms, cloning and its applications in gene therapy. The basic molecular mechanisms regulating ageing, stem cell biology and their use in modern cellular therapies for diseases and neoplasms are described. Finally, the basic principles of tumorigenesis, oncogenes and tumor suppressor genes, the role of cancer stem cells in cancer transformation and new cancer therapies are discussed.

Upon successful completion of the course the student will be able to:

– Have an understanding of the biology of self-replicating macromolecules and their interactions with proteins

– Have knowledge of the basic functional structures of the human genome

– Have knowledge of basic mechanisms of genetic stability perturbation (carcinogenesis, hereditary cancer/ageing)

– Knowledge of modern molecular biology methods and their applications in the diagnosis and treatment of diseases and malignancies

– Able to distinguish the affected individual from the normal individual by interpreting the results of laboratory tests (e.g. PCR)

– Use a variety of laboratory instruments and equipment (pipettes, PCR apparatus, nucleic acid electrophoresis devices, etc.)

The course material includes the following sections:

– Molecular Biology in Medicine

– Molecular Biology and Personalised or Precision Medicine, Results of the Human Genome Project.

– Genome structure and organization

– Nucleosomes (structure, organisation, function), Interactions with proteins in centromeres and telomeres, Chromatin structure in the interphase nucleus, Organisation of genes in the genome, Types of genes, Repeat sequences, Scattered repeats, Transposable elements, Non-coding DNA.

– Physicochemical properties of nucleic acids

– Nucleic acids, DNA/ RNA structure, DNA/ RNA differences/similarities, Types of RNA, Transcriptome

– DNA replication

– DNA polymerases, Replication errors and proofreading activity of DNA polymerase, Replication machinery and its constituent enzymes (primases, helicases, topoisomerases), Telomers and telomerase.

– DNA damage and repair

– Types of DNA damage, Mutagenic agents (radiation, chemical agents), Basic mechanisms of damage repair (NHEJ, homologous recombination), Damage to repair mechanisms: the example of melanchromatic dry skin and classical progeria syndromes.

– RNA transcription and processing

– Structure and function of RNA polymerase; RNA species and RNA polymerases; Differences between eukaryotes and prokaryotes in transcription; Mechanism of transcription; Transcription factors (general, specific); Transcription of mitochondrial genes; Processing of RNA molecules; Splicing and alternative splicing: alternative splicing in familial hypercholesterolemia; MRNA degradation and exonuclease activity.

– Translation

– Transfer of genetic information from RNA to proteins, Redundant genetic code and the wobble or instability hypothesis, Translation initiation factors (elFs), Controlled protein degradation-structure and function of the ubiquitin-proteasome system and diseases: Autoimmune diseases, Parkinson’s disease, viral infections, cancer, Lysosome-dependent protein degradation, Post-translational modifications of proteins, RNA and origin of life, RNA-DNA evolution.

– Regulation of gene expression in prokaryotes

– Response to exogenous segments and modification of gene expression, Molecular switches, Basic structure and function of operon in prokaryotes (activators and repressors), Regulation of the lactose operon and the action of CAP protein, Regulation of the tryptophan operon and the concept of co-repressor, Secondary mechanism of operon regulation-transcriptional attenuation.

– Regulation of gene expression in eukaryotes

– Checkpoints in the transfer of genetic information from DNA to proteins, Transcriptional regulators and regulatory sequences; nuclear steroid hormone receptors and tamoxifen action, Activators and repressors of expression, Enhancers, Combinatorial control of gene expression and development.

– Epigenetic regulation of gene expression

– DNA methylation, histone modifications, large and small non-coding RNA molecules, Dysfunction of epigenetic mechanisms and disease pathogenesis, Small RNA interference (RNA interference/RNAi).

– Mutations and polymorphisms

– Types of mutations and their involvement in diseases: the example of sickle cell anaemia and cystic fibrosis, Polymorphisms (SNPs, STRs, VNTRs, CNVs), Polymorphisms as predisposing factors of diseases: Apolipoprotein E genotypes and Alzheimer’s disease, Polymorphisms and forensic DNA fingerprinting, Polymorphisms of mitochondrial DNA, Polymorphisms and drug response: example of polymorphisms in cytochrome P450 isoenzyme and antithrombotic therapy.

– Stem cells and tissue homeostasis

– Types and properties of stem cells, Method of reprogramming differentiated cells and production of induced stem cells, Cell cycle of adult stem cells, Damage to tissue signalling and induction of adult stem cell proliferation, Stem cells and cell therapy of diseases: Neurodegenerative and cardiovascular diseases, spinal cord injury, cancer, etc. α).

– Molecular mechanisms of ageing of cells and organisms

– Ageing as a response to damage accumulation, Milestones of ageing, Ageing as a barrier mechanism in carcinogenesis and as a risk factor for carcinogenesis.

– Molecular Biology Technologies

– Polymerase chain reaction (PCR) and real-time polymerase chain reaction (RT-PCR), Southern blot, Northern blot, Sanger method, Role of molecular biology technologies in biomedical research and their importance in clinical practice – molecular diagnostics.

– New technologies and applications in disease diagnosis

– Next generation sequencing (NGS), Comparative analysis of genomes, Gene identification and function prediction, DNA microarrays and RNA-seq, Applications in the diagnosis of genetic diseases (cystic fibrosis etc.) and malignancies (solid tumours and haematological neoplasms).

– Modern recombinant DNA technology

– Recombinant DNA technology, Genetic engineering, DNA manipulation and analysis techniques (restriction nucleases).

– Cloning

– DNA cloning in bacteria – use of plasmids and transformation of bacteria, DNA library generation (genomic library and cDNA library).

– Experimental animals as model organisms

– Model organisms (Drosophila, C. elegans, mouse), Ways of creating transgenic, conditional transgenic and knock out mice, Contribution of transgenic organisms to elucidate biological processes in the aetiopathogenesis and treatment of diseases. Animal models of human diseases (Huntington’s disease, progeria, Zucker rats and obesity).

– Evolution of genomes

– Ways of generating genetic diversity, gene change, Natural selection pressure, evolution: lactose intolerance, The paradigm of horizontal gene transfer and antibiotic resistance, Homologous genes (highly conserved genes), Genetic drift, Parts of conserved synteny, Inadmissible and intolerable changes in the genome; purifying selection.

– Carcinogenesis

– Mechanisms of carcinogenesis, Oncogenes and tumour suppressor genes: the example of retinoblastoma, Loss of heterozygosity and microsatellite instability (MSI), Cancer stem cells and cancer transformation, New therapies in cancer.

Laboratory Exercises (Two modules (Part A and B) conducted in 5 laboratory exercises)

Part A : Diagnosis of sickle cell anaemia by Restriction Fragment Length Polymorphisms (RFLP)

– Use of microplate – Isolation of genomic DNA from peripheral blood using columns

– Amplification of part of the beta-globin gene by polymerase chain reaction (PCR) technique

– PCR product segmentation using restriction endonucleases

– Preparation of agarose gel and electrophoresis of digestion products in the gel

– Evaluation of the result; diagnosis of sickle cell anaemia

Part B: Serological Diagnostic Method-ELISA

– Basic Principles of Immunological Response – Antibodies/Chemical Immunity

– ELISA (Enzyme-Linked ImmunoSorbent Assay)-Detection of antibodies (e.g. against measles virus)

• Course notes
• Molecular Cell Biology, Lodish H et al. [EUDOXOS 122091150]
• Genomes – modern research approaches, : Brown T. A. [EYDOXOS 122074091]

Nature Reviews Molecular Cell Biology, Molecular Cell, Molecular Biology and Evolution, Molecular Aspects of Medicine