|
|
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...
|
|
|
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...
|
|
|
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...
|
|
|
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.
|
|
|
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
|
|
|
The analysis of autotransfusions carried out in the FN Motol
Šámalová, Martina ; Linhartová, Eva (advisor) ; Závadová, Žaneta (referee)
The theoretical part of the bachelor's thesis includes information connected with the autologous taking, the production of the autologous transfusion preparations and the protection of quality of the autotransfusion, also basic information about blood groups, which are the integral part of the transfusion medicine. The practical part is divided into three main parts. The first part deals in more detail with the issue of blood taking for autotransfusion and with the examination connected to autologous blood. The second part contains the statistical figures about the autotransfusions carried out in the Blood Bank Department of FN Motol. It examines the quantity of taken and expired transfusion preparations of patients indicated for the autologous blood taking from the orthopaedics department in total knee and hip joint endoprosthesis and from the urology department of pacients before prostate operations and transformation of gender from male to female. The third part consists of the quality control and the protection of quality of the autologous transfusion preparations. The practical part is based on statistical materials provided by the Blood Bank Department of FN Motol of the year 2011.
|
|
|
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...
|
|
|
Origins of vertebrate hematiopoiesis
Svoboda, Ondřej ; Bartůněk, Petr (advisor) ; Divoký, Vladimír (referee) ; Živný, Jan (referee)
(ENGLISH) Hematopoiesis is dependent on the actions of hematopoietic stem cells (HSCs). This process is tightly controlled through a complex array of extrinsic and intrinsic factors. Even though the hematopoiesis seems to be well conserved across the disparate vertebrate animals, erythroid and thrombocytic differentiation have changed during the evolution of mammals. Specifically, adult mammalian red blood cells have the unique feature of being enucleated, and mammalian thrombocytes are not individual cells, but fragments of megakaryocytes, instead. It is likely that these enhancements provided a survival advantage to early mammalian species; however, they also bring up the question of evolutionary origin of these cells that studied using zebrafish (Danio rerio) model. First, it was necessary to generate a toolbox of a recombinant cytokines and optimized culture media that allowed us to manipulate zebrafish hematopoietic cells ex vivo in liquid and clonal cultures. Interestingly, teleost species underwent an extra duplication event during their evolution and as a result, two copies (paralogs) of some of the genes are present in zebrafish. This was also the case for majority of the cytokines from our toolbox and here, we provide functional characterization of these paralogs. Strikingly, our results...
|
|
|
Comparison of microscopic and instrumental results of individual blood cell types with a focus on platelets
DRÁBOVÁ, Michaela
The aim of my thesis is to determine the number of platelets with different methods; then to determine the cause of the measured differences in the number of platelets, followed by determination of algorithm for reliable counting of platelets. In specific situations, the instrument determination of the levels of platelets may be significantly different from the real situation. The actual number of platelets can be obtained by using alternative methods. Microscopic assessment and determination of the number of platelets in sodium citrate are considered as alternative methods. Hypothesis I. When using alternative methods, different results are obtained in part of the population. Hypothesis II. These alternative methods help us to distinguish between true and false thrombocytopenia. The theoretical part deals with the formation, structure, physiology, function, metabolism and characteristics of platelets. The term of thrombocytopenia is defined. Quality control of laboratory analyzes is mentioned. Different anticoagulant products used in hematology are described. There are also described techniques by which platelets can be determined and interference associated with the measurement of blood cells. The practical part includes the processing of biological material in a test-tube with the purple cap containing anticoagulant EDTA (from this test-tube a complete blood count is provided), while a test-tube with the blue cap contains anticoagulant sodium citrate (this test-tube is only for re-determination of the platelets count). Analysis of biological material was performed by the analyzer Coulter LH 750, Beckman Coulter Inc. The Coulter method measures the size of blood cells due to changes in electrical resistance. Manual processing of biological material was performed using a light microscope Carl Zeiss Inc., the total magnification for observation of platelets is 400 times. Number of platelets is evaluated in Bürker chamber in 20 rectangles. Solution of procaine is used to stabilize and highlight the platelets. In the period of 23rd April - 22nd May 2012 at the department of Clinical Hematology, Nemocnice České Budějovice a.s., I examined the platelets count in two different anticoagulants by using different measurement techniques. This comparison is important because if the difference in the number of platelets found in EDTA anticoagulant compared with sodium citrate is higher than 20 %, the samples could be suspected of pseudothrombocytopenia. The measurement included fifty samples which were selected already with some suspicion according to following two criteria: the platelets count had to fall below 100 x 10^9/l and at the same time we had received analyzer report of the presence of clumps. The measured values of platelets were statistically processed and they are shown in charts and graphs in my thesis. For a better idea there are also pictures - especially histograms showing some changes in the number and volume of blood cells. This research has demonstrated the occurrence of the phenomenon PTCP in 0,052% cases. The difference between the techniques was clearly evident. Microscopy does not reach such accurate results as analyzers. Quantity of samples and speed of implementation can not be compared. Nowadays microscopy is used as an auxiliary technique in those cases where the analyzer is too affected by imperfections (for example formation of clumps) that it gives false platelets count. The disadvantage is the time delay from the sampling to the possibility of counting. By alternative methods we reach eventual verification or modification of first results (for example repeated sampling to another anticoagulant).
|
|
| |
guest ::
