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Anti-hemostatické účinky serpinů z klíštěte \kur{Ixodes ricinus}
EDEROVÁ, Monika
Antihemostatic effects of four recombinantly produced serpins of the tick Ixodes ricinus (IRS 2, IRS 3, IRS 5 and IRS 8) were tested using a method of flow cytometry. Fluorescently labeled antibodies against platelet surface molecules were used to determine the difference in platelet aggregation and activation after incubation of samples with specific serpin. To see how serpins affect coagulation pathway, the prothrombin time, activated partial thromboplastin time and thrombin time tests were performed.
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Preparation, characterization and testing of blood derivatives for applications in regenerative medicine
Sovková, Věra ; Amler, Evžen (advisor) ; Kolářová, Hana (referee) ; Vránová, Jana (referee)
Platelet products can be used, thanks to the broad range of bioactive molecules, either as a supplement for cell cultering in vitro alone or for development of cell- or cell-free scaffolds in diverse fields in regenerative medicine. The aim of this study was to prepare several types of platelet products. The concentration of selected molecules were observed. Subsequently, these products were tested with cell cultures in vitro alone or in combination with nanofibres scaffolds prepared by electrospinning or centrifugal spinning. It was found out, that platelets products contains chemokine RANTES and growth factor PDGF in the highest concentrations. It was further discovered the content of pro and antiinflammatory in terleukins and other growth factors. Platelet lysat in concentration 7% is sufficient to replace FBS in keratinocytes and fibroblasts cultures. In the other experiments, platelets in different concentrations were adhered to the scaffolds prepared by electrospinning and centrifugal spinning. Thus prepared scaffolds promote the proliferation and viability of all tested cell types in dose-dependent manner. In the last experiment, the individual components of platelet concentrate were separated and characterized. Their effect to the cell culture were tested. It was examinated the synergic...
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Comparison of control measurements on different types of analyzers used in ÚKBLD CHLTC of VFN in Prague
Koblasa, Vladimír ; Kvasnička, Jan (advisor) ; Kozák, Tomáš (referee)
Koblasa,Vladimír - Comparison of control measurements on various types of haematology analyzers used by ÚKBLD CHLTC at University Hospital in Prague First Faculty of Medicine, Charles University in Prague, Praha 2, Kateřinská 32 Head of the work: prof. MUDr. Jan Kvasnička, DrSc. Supervisor -consultant: Mgr. Ivana Malíková Blood cell count is essential testing method in hematology, where it is necessary to ensure properquality control. Aim of this study was to compare the results of measurement control materials with defined parameters and the same samples at different haematological analyzers to obtain evidence for the expression of measurement uncertainties. There are used more types of blood analyzers in ÚKBLD CHLTC at University Hospital in Prague, which operate on different principles. For comparsion were selected analyzers using the impedance working principle, where individual blood cell passes between two electrodes controlled by low voltage. Variation of this voltage is recorded and accurately defined for each type of blood cells. It was also chosen analyzer that works with optical detection. Analyzer illuminates the individual blood cell by light beam. A cell that enters into the path of light rays, reduce its optical density incident on the photocell. The change of the light density...
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Production and use of blood products in the Regional Trutnov Hospital.
Rosová, Valérie ; Červená, Gabriela (advisor) ; Honegrová, Barbora (referee)
Title of Diploma Thesis: Production and use of transfusion products in the District Hospital Trutnov. Aim of the work: The aim was to evaluate the production of transfusion products (TP) in the period 2014-2019 at the Transfusion and Hematology Department (THO) in the District Hospital Trutnov and their use for hemotherapy by individual medical disciplines. Futhermore, to compare the production of TP within the Czech Republic together with selected European countries. The incidence of post-transfusion reactions and other adverse events during the reporting period is also included. Material and methods: The methodological part describes the standard procedures of pre- transfusion examination using the column agglutination method from the company Grifols, S.A., laboratory examination of donors and transfusion products manufactured at the Transfusion Department Trutnov. Conclusion: Despite all the recommendations and efforts to reduce the use of transfusion products, this condition has not been proven both in our department, within the Czech Republic and in Europe. This work recommends deleucotization of transfusion products for the national and European part, unification of procedures for transfusion in a given disease, mandatory introduction of NAT testing to minimize the risk of transmission...
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Whole Blood - New Blood Component Containing Platelets
Nováková, Kateřina ; Červená, Gabriela (advisor) ; Honegrová, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Bc. Kateřina Nováková Supervisor: RNDr. Gabriela Červená, Ph.D. plk. MUDr. Miloš Bohoněk, Ph.D. Title of diploma thesis: Whole blood - New Blood Component Containing Platelets Background: The aim of this thesis was to evaluate in vitro quality parameters and haemostatic function of leucodepleted WB and their comparison with non-leucodepleted WB. The same way the quality parameters (RBC, PLT, HB and hemolysis) of erythrocyte blood component were measured and compared in day D14, D21, D35 and D42. Methods: WB collected from 30 healthy group A donors was divided into two groups - leucodepleted (LWB), using in-line platelet-sparing filters and collected to a blood bag system IMUFLEX® WB-SP (Terumo BCT, USA) and non-leucodepleted WB (NLWB), colletected to a blood bag system CompoFlex® Single System (Fresenius Kabi, Germany). Both groups were stored at 42řC for 14 days and following parameters were measured in days D0, D1, D3, D5, D10 and D14: WBC, RBC, PLT, HB, hemolysis, pH, TEG, FVIII, TT, PT, aPTT, aggregometry, concentration of PF4 and sCD40L (ELISA). Moreover, in days D0, D7 and D14 was measured the level of expression of platelet activation marker CD62P (P-selectin), CD42b, CD61 by...
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Role of Rnf207b in zebrafish hematopoiesis
Vondráková, Zuzana ; Bartůněk, Petr (advisor) ; Živný, Jan (referee)
Hematopoiesis is the process of proliferation, differentiation and self-renewal of hematopoietic stem cells. Regulation of hematopoiesis is a complex process, which takes place on many different levels and is directed by many signals. RNF207 is one of the perspective genes chosen based on a screen in chicken model, where obtained data show its role in hematopoiesis. The aim of this work was to confirm the role of rnf207b as a new regulator of hematopoiesis in Danio rerio and to find out on which level of hematopoiesis is active. Danio rerio is an excellent model to study the function of genes in vivo, thanks to the easy manipulation of genetic expression and wide range of phenotypes during the development. To study the effect of rnf207b in hematopoiesis of Danio rerio we performed the knock-down of this gene by microinjection of morpholino oligonucleotides into one cell stage embryos. In these injected fish, we saw the effect in both thrombocyte and erythroid lineage, suggesting that rnf207b could be a regulator at the hierarchical level of progenitors or even more upstream. The results of developmental and tissue specific expression analysis then show that expression of rnf207b begins as early as 18 hpf, at the time of primitive hematopoiesis. Although rnf207b is expressed in the kidney (an...
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Preparation, characterization and testing of blood derivatives for applications in regenerative medicine
Sovková, Věra ; Amler, Evžen (advisor) ; Kolářová, Hana (referee) ; Vránová, Jana (referee)
Platelet products can be used, thanks to the broad range of bioactive molecules, either as a supplement for cell cultering in vitro alone or for development of cell- or cell-free scaffolds in diverse fields in regenerative medicine. The aim of this study was to prepare several types of platelet products. The concentration of selected molecules were observed. Subsequently, these products were tested with cell cultures in vitro alone or in combination with nanofibres scaffolds prepared by electrospinning or centrifugal spinning. It was found out, that platelets products contains chemokine RANTES and growth factor PDGF in the highest concentrations. It was further discovered the content of pro and antiinflammatory in terleukins and other growth factors. Platelet lysat in concentration 7% is sufficient to replace FBS in keratinocytes and fibroblasts cultures. In the other experiments, platelets in different concentrations were adhered to the scaffolds prepared by electrospinning and centrifugal spinning. Thus prepared scaffolds promote the proliferation and viability of all tested cell types in dose-dependent manner. In the last experiment, the individual components of platelet concentrate were separated and characterized. Their effect to the cell culture were tested. It was examinated the synergic...
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Intelligent nanofibres functionalized with growth factors and blood derivatives for dermatology applications
Vocetková, Karolína ; Amler, Evžen (advisor) ; Rosina, Jozef (referee) ; Arenberger, Petr (referee)
Platelet derivatives are an attractive source of natural growth factors and they are widely used in various tissue engineering and regenerative medicine applications. The aim of this study was to optimize cell culture conditions using platelet lysate and to develop platelet-functionalized fibrous scaffolds as a controlled drug delivery system for native growth factors. Fibrous scaffolds were prepared by electrostatic and centrifugal spinning of PCL and they were functionalized by the platelets by surface adhesion or their encapsulation using emulsion spinning techniques. The cell culture study determined the 7% platelet lysate to be the optimum concentration as a medium supplement in keratinocyte and fibroblast culture. Additionally, following surface adhesion of the platelets to PCL electrospun nanofibres, the platelets were activated due to their contact with the nanofibre nanotopography, resulting in formation of fibrin network. Fibrin served as a reservoir of the growth factors, prolonging the half-time of EGF release to 1.7 days. Such platelet-functionalized samples fostered proliferation of keratinocytes, fibroblasts and melanocytes. Furthermore, adhesion of platelets to centrifugally spun nanofibrous scaffolds resulted in almost two-fold increase in the amount of immobilized platelet-derived...
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Historie a současnost léčby idiopatické trombocytopenické purpury
ŠIMKOVÁ, Veronika
The bachelor paper on The Past and Present of Idiopathic Trombocytopenic Purpura Therapy explores this disorder with particular emphasis on modern methods of treatment. It describes its origin and classification; its causes and incidence in the population; its diagnostics and pathogenesis. The text also includes a questionnaire related to its treatment in the South Moravian Region as well as cause studies of individual patients.
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Haematopoiesis in Sea lamprey
Kovář, Martin ; Bartůněk, Petr (advisor) ; Živný, Jan (referee)
To find out if the haematopoietic system is common feature of vertebrates, we decided to examine haematopoiesis in a sea lamprey (Petromyzon marinus). All blood cells arises from the haematopoietic stem cells in higher vertebrates. We assume that this is common for the higher vertebrates and a jawless vertebrates, but nobody was interested in the jawless haematopoiesis since 1970. Using a reverse genetic, we identify homologues of important hematopoietic of higher vertebrates in transcriptome of the sea lamprey with emphasis on important receptors or transcription factors, because they can be used as the specific markers of different blood cells and their progenitors. Then we use those sequences for cloning, expression measurements and other work. We picked up sea lamprey as model organism because its unique phylogenetic position, important foe evo-devo studies, but also because lack of elementary knowledge about sea lamprey haematopoiesis. Key words: Petromyzon marinus, haematopoiesis, HSC, evo-devo
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