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Biotechnology in the world and the Czech lands
Pěknicová, Jana
Biotechnology in the world and the Czech lands Biotechnology is any technology that uses biological systems, living organisms, or parts of a particular production or their conversion or other specific applications. The lecture should show a brief overview of the biotechnology industry in the world, in the Czech lands, in the Institute of Biotechnology CAS, v. v. i., and in the project BIOCEV: 1. Biotechnology in the world • Beginning of biotechnologies • Milestones • Application in medicine • Gene therapy • Genetic modification (science, agriculture, industry) 2. Biotechnology in the Czech lands • Czech biotechnology: milestones 3. Institute of Biotechnology CAS, v. v. i. • Research and outputs into practice 4. BIOCEV – Biotechnological and Biomedicine Centre of the Academy of Sciences and Charles University in Vestec Establishment of centre of excellence as part of the European scientific area guaranteeing the development of modern biotechnology and medicine.
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Biochemical methods as tool for study of reproductive proteins
Postlerová, Pavla ; Zigo, Michal ; Pohlová, Alžběta ; Jonáková, Věra
Study of molecular mechanisms in reproduction is essential for the understanding of this outstanding process. Our lab studies proteins secreted by reproductive organs and sperm using various biochemical methods for a long time. We have expertise in protein extraction from spermatic cells using different approaches, and by kits for proteins from the sperm surface and distinct subcellular compartments. The proteins of reproductive organ fluids are separated by chromatographic methods, such as size exclusion chromatography, high-performance liquid chromatography with reverse phase (RP-HPLC) and affinity chromatography on matrices with various ligands. Proteins are subjected to SDS- or 2D-electrophoresis for their characterization and comparison of various extraction methods, different mammalian species, and sperm in different functional development. Electrophoretically separated proteins may be transferred onto nitrocellulose membrane (Western blot) for antibody detection or binding studies with lectin-labelled ligands (lectins, polysaccharides, zona pellucida glycoproteins). We use immunoprecipitation method with specific antibody for protein determination followed by the MALDI identification. Proteins are localized by immunofluorescent techniques on/in spermatic cells and tissue sections of reproductive organs. Isolation of proteins from reproductive tissues and fluids, and the antibody detection is crucial for the studying of reproductive protein origin.
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Expression of selected proteins in sperm in men with normal and pathological spermiograms using monoclonal antibodies
Pěknicová, Jana ; Čapková, Jana ; Dorosh, Andriy ; Margaryan, Hasmik ; Kubátová, Alena ; Děd, Lukáš
Recent studies show that infertility in human populations it affects an estimated 15% of couples of reproductive age. Male infertility is the primary cause for 60% of these cases. For these reasons, we analyzed the acrosomal and sperm surface proteins in men with normal and pathological spermiograms. We found that intra-acrosome proteins: TERA (Transitional endoplasmic reticulum ATPase), GAPDHS (Sperm Glyceraldehyde-3-phosphate dehydrogenase), and PRKAR2A (C-AMP-dependent protein kinase II, PRKAR2A), which can be identified using our monoclonal antibodies, are different express in healthy men and men with astenozoospermia (with reduced sperm motility), and with a significantly reduced expression in the astenozoospermia. These proteins are involved in energy metabolism and apoptosis of the cells, and some of them in the sperm-egg interaction; therefore, they have an important role in reproduction. On the other hand, there were no statistically significant differences in the expression of surface proteins (Appolipoprotein J (Clusterin) and Semenogelin). Our findings show that astenozoospermia as a complex disorder of the semen is often combined with other pathological conditions that are not diagnosed by the semen analysis. Therefore monoclonal antibodies are so suitable instrument for the detection of proteins associated with the pathology of the sperm in the semen with low sperm motility. In general, monoclonal antibodies against the sperm proteins are an appropriate tool to detect sperm quality in reproductive medicine.
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