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Analysis of antimicrobial peptides by capillary electrophoresis
Ehala, Sille ; Niederhafner, Petr ; Čeřovský, Václav ; Řezanka, P. ; Sýkora, D. ; Král, V. ; Kašička, Václav
Capillary electrophoresis was used for the purity degree determination of new antimicrobial peptides: octadecapeptide melectin and tetradecapeptide mastoparan PDD-B, and their synthetic analogues. Besides, four semiempirical models correlating electrophoretic mobilities of these peptides with their charge and size (relative molecular mass) were applied to predict their probable structures in solution. Additionally, capillaries coated with gold nanoparticles were employed for the separation of mixtures of antimicrobial peptides.
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Model Activities of the Integrated Rescue System Forces in Joint Actions from the Point of View of the Emergency Medical Service
KRÁL, Václav
A set of documents, called the Index of Model Activities of the Integrated Rescue System Forces was compiled to improve coordination of these forces. The Index describes organizational procedures of the IRS forces during ten cases of emergency. However, are members of basic IRS forces familiar with these documents? Are the IRS workers aware of procedures and needs of the Emergency Medical Service (EMS) staff? The objective of this thesis is to survey views and findings of the IRS workers regarding the above mentioned issues, evaluate possibilities to improve knowledge of these issues.
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Verification of the effect of rations on goats feed intake and on milk composition
KRÁL, Václav
Goat's milk has many great nutritional properties, which arise from its unique composition. The milk composition is however depending on the composition of the feed diet. The aim of this work is to determine how the diet affects the quantity of milk yield and composition. We also made observations to determine how selected herbs affect the intake of feed rations with goats. The experimental part is divided into three parts. In the first part, we evaluated the impact of selected herbs to income ration with goats. We found that the addition of perennial ryegrass (Lolium perenne) increases the amount of food taken. Conversely, an addition of meadow pea (Lathyrus pratensis) decreases the amount of food intake. In the second part, we tested the ribwort plantain (Plantago lanceolata) effect on the milk composition. We found that an addition of this herb in the ration slightly increases daily milk yield. In the third section we monitored a herd of goats on grassland. We evaluated influence of the composition of pasture vegetation and the addition of oats on milk yield and composition. Our conclusion is that the composition of the pasture had a significant impact not only on the basic components of milk, but also on the representation of individual groups of fatty acids. The adding of oats decreases the influence of grazing.
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Etické aspekty ošetřování nespolupracujících pacientů v rámci záchranné služby
KRÁL, Václav
Emergency health service is a specific part of health care and in connection with ethical approach to non-cooperating patients many questions appear: To what extent is acceptable to force the patient to cooperate and to what extent is it necessary and important to leave the patient the right to decide on his or her health in life? Is it in accordance with ethics to tell the patient a white lie on his or her state so that he or she would be willing to cooperate? The aim my thesis is to find out which approach is ethically acceptable when treating patients who do not cooperate. Further I wanted to learn about the opinions of rescue workers on ethical dilemmas, which I described above.
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Molecular cloning, E.coli expression and purification of SCFV antibody fragments of diagnostic/therapeutic interest
Král, Vlastimil ; Fábry, Milan ; Hořejší, Magdalena ; Závada, Jan ; Sedláček, Juraj
We describe molecular cloning, expression, purification and properties of two single chain antibody variable fragments (scFv) of potential diagnostic use, namely scFv M75 and scFv Tu-20. The former scFv is derived from a monoclonal antibody M75 specific for a cell surface protein MN/CA IX, strongly associated with many types of human carcinomas. The latter scFv is derived from a monoclonal antibody TU-20 specific for neuronal beta-III-tubulin.
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Crystallographic study of an anti=carbonic anhydrase IX monoclonal antibody M75
Štouračová, Renata ; Závada, Jan ; Závadová, Zuzana ; Pastoreková, S. ; Brynda, Jiří ; Fábry, Milan ; Král, Vlastimil ; Hořejší, Magdalena ; Sedláček, Juraj
Carbonic anhydrase IX (CA IX) is a cell surface protein, strongly associated with certain types of human carcinomas. Structural study of a CA IX-binding monoclonal antibody (mAb) M75, complexed with its epitope peptide may contribute toward elucidation of the role of CA IX. Monoclonal antibody M75 was obtained and proved to react excellently with native and denaturated CA IX. Using synthetic oligopeptides, the epitope of mAb M75 was localized in the proteoglycan domain of CA IX, in the region of a tandem repeat and identified as amino acids PGEEDLP. The Fab fragment was obtained by papain cleavage. We obtained crystals of free Fab M75 and Fab M75 complexed with two different epitope peptides. The data set for Fab M75 was collected and the structure solving is underway.
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