Cell Biology of Neurodegenerative Diseases 1. Prion diseases Prion disease: a new entity.
Its significance in biomedical and basic research. Classification of prion diseases: human and animal forms. Brief characterization of scrapie, bovine spongiform encephalopathy BSEkuru and Creutzfeldt-Jakob disease. Infectivity, as a unique phenomenon. General overview of cell, tissue and organ culture systems: in vitro methods, their advantages and disadvantages, conditions and levels of their applications.
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The most important conditions and factors for in vitro survival, safety measures, the analyis and evaluation of the results, conclusion to in vivo processes. Perfusion systems: in situ and isolated-perfused organ systems and organs, main types of arrangements constant flow-rate and pressure, single-pass and recirculation systemstechnical ground, perfusion fluids. Some examples of organ perfusion systems: liver, kidney, heart, pancreas.
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Organ cultures without perfusion. Embryonic and non-embryonic cultures, organitypic growth, solid, semisolid and liquid media, the problems of oxigenation, Developmental biology II An overview of body structure of the model organism Caenorhabditis elegans. Handling and propagation of worms. Cell lineage, stages of embryonal and postembryonal development.
Investigation of normal and developmental mutant worms Gametogenesis and localization of morphogens in the oocytes of Drosophila melanogaster. Determination of body axes and segments, introduction to stages of embryonal development. Anatomy of larvae, specificities of larval and imaginal tissues.
Hormonal and genetic control of metamorphosis and organogenesis. Fertilization and embryonic develoment of zebrafish. An overview of molecular mechanisms that control fish development.
Larval organs and larval development. Formation of the skull and adult organs. Pattern of morphogens in frog oocyte. Developmental genetics 1.
Mutant analyis in nematodes C. Epistatic analysis in worms C. Double mutant analysis. Recessive epistatis, dominant epistasis, complementer inheritance, additive genetic interaction. Dissecting genetic pathways in C. Recessive epistasis.
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Analysis of the core apoptotic world journal of diabetes predatory death pathway in C. Epistasis analysis. Analysis of the Gametogenesis: conversion of primordial germ cells into gametes. Mitosis, meiosis, polar bodies.
Chromosomal abnormalities: 1. Oogenesis: maturation of oocytes before birth — primary oocytes, primordial follicles.
Maturation of oocytes at puberty — primary, secondary and Graafian follicle. Ovulation and corpus luteum. Histology of ovary.
Spermatogenesis: primordial germ cells and supporting cells in sex cord world journal of diabetes predatory newborn boy. Puberty: development of seminiferous tubules, spermatogonia, spermatocytes and spermatids.
Immunocytochemistry Theory: Methods of protein separation from biological samples.
Planning and cloning of an antigen of interest. Production of mono- and polyclonal antisera. Antisera characterization and antibody labelling.
Preparation of samples: collection, fixation and embeding of different biological samples. Antigen retrieval technics and signal enhancement.
Bases of in situ hibridization and FISH methods.
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Practice: 1. Immunohistochemistry Localization of different cytoskeletal proteins cytokeratin, tubulin and actin in the cells of fibroblast culture by using two-steps immunohistochemistry. Fluorescein microscopy and digital microphotography.
Localization of a cuticular protein on paraplast-embedded sections from Manduca larvae by using enzyme -labelled second antibody.
Introduction — Summary of mammalian morphogenesis, germ layers and their derivatives mesodeum, somites, lateral mesoderma, head mesoderma, neural plate, neural crest.
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Pattern formation, secondary induction - interaction between epithelial and mesenchymal tissues. The common steps in the formation of parenchymal organs: generation of branches in the insect tracheal system, the mammalian kidney and airway system.
Hox-genes — pattern formation along the antero-posterior axis, RA in developmental malformations. Morphogenesis of the mammalian viscerocranium: pharyngeal pouches, determination of the polarity, position and identity of the branchial arches Bmp-7, Fgf-8, Pax1 and Shh Molecular evolution 1.
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Fundamentals of molecular evolution: Overview: genes and gene structure; genetic code; mutations and their consequences: changes in the DNA sequence. Molecular evolution of amino acid sequences: amino acid differences and proportion of different amino acids; amino acid substitution matrix. Molecular evolution of DNA sequences: nucleotide differences between sequences; nucleotide substitution in DNA sequences: Jukes-Cantor's one parameter model, Kimura's two parameter model.
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Sequence alignments for molecular phylogenetics: pairwise alignments; local and global alignments; substitution matrices; dot-plot, dynamic programming algorithm. Multiple alignments: extension of dynamic programming; hidden Markov models; genetic algorithm in multiple alignment.