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Molecular mechanism of autosomal dominant polycythemia
Berková, Linda ; Láníková, Lucie (advisor) ; Kapráľová, Katarína (referee)
All red blood cells, erythrocytes, originate in bone marrow. The process of their differentiation and maturation is called erythopoiesis and is regulated through hormone erythropoietin (EPO), which functions as a stimulatory factor for erythropoiesis. EPO is produced in kidney and its production is regulated by oxygen supplementation. EPO is transported to bone marrow via blood vessels. Chronic overproduction of erythrocytes leads to disease called polycythemia. Polycythemia may be diagnosed for example by measurement of haematocrit or haemoglobin concentration in blood. EPO level may or may not be increased. Patients suffering from polycythemia may or may not have any symptoms. It depends on manifestation level of the disease. The most common symptoms are higher blood pressure, headaches, dizziness, swelling and epistaxis. Recently, the most common treatment of polycythemia is phlebothomy. The aim of this master thesis is to unravel the role of a newly described mutation, which was found among members of one family suffering from polycythemia with increased EPO level. It is single substitution mutation -136 G > A in 5' UTR region of EPO gene. The clones of EPO producing cell lines bearing this mutation were prepared using CRISPR/Cas9 technology. Several experiments performed not only on those cell...
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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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Application of Haematological Methods in Zoological Studies
Poplová, Jitka ; Vinkler, Michal (advisor) ; Krulová, Magdaléna (referee)
Haematological methods are widely used in zoological researches. Unfortunately, especially in ecological studies there is often much imperfection in techniques adopted, and misinterpretation of results is also common. This is particularly the case of methods serving to assessment of the peripheral blood cellular composition. In this thesis I therefore decided to describe basic haematological methods, their mechanisms, advantages and disadvantages. I also highlighted possible application potential in several less widely utilised haematological techniques. In the practical part of this thesis I have shown that the examination of immature erythrocyte differential count may serve as a meaningful indicator of health state in birds.
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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.
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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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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...
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Variabilityin egg shell pigmentation of great tit clutches
Kratochvílová, Anna ; Svobodová, Jana (advisor) ; Veronika, Veronika (referee)
Eggshell coloration in bird clutches shows a striking variability within species which has not yet been fully explained. Its character is determined by the deposition of two main pigments: blue-green biliverdin is responsible for the background colour while brown-red protoporphyrin is associated with the dark spot pattern. Considering the role and matabolism of these two pigments in the avian organism it was suggested that they might have an important impact on the oxidative stress levels and their deposition into the eggshell is therefore suggested to possibly reflect the body condition of laying females during the laying periods. This is also the basic assumption for the signalling hypothesis of eggshell coloration which predicts the eggshell appearance to reflect the female body condition and individual health status or directly indicate aenemia in birds (aenemic hypothesis). Here it was tested whether there is a relationship between parameters representing the body condition of female great tits (Parus major) and the characteristics of protoporphyrin maculation of their eggs. Female body condition was expressed by standardized body weight and the selected haematological parameters (heterophil to lymphocyte ratio and relative immature erythrocyte counts). Eggshell coloration was characterized by spot colour (hue, saturation and brightness), spot counts and the reflectance of the background colour. It was found that females with higher heterophil to lymphocyte ratio laid eggs with more spots and higher reflectance of the background colour in some nesting seasons. Eggs with more spots on its surface were also laid by females with higher immature erythrocyte counts. These results suggest that more intensive deposition of protoporfyring into the eggsell can indicate worse body condition of the laying females. On the other hand, it was also found that heavier females laid eggs with lower reflectance of the background colour. Despite this discrepancy, these results are not inconsistent with the pressumptions of the signalling hypothesis and they partially support the aenemia hypothesis. However, the above mentioned dependencies were inconsistent among the nesting seasons, indicating that further important determinants must be considered in the avian eggshell coloration.
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Detection of blood elements
Kadlček, Václav ; Kolář, Radim (referee) ; Lamoš, Martin (advisor)
Thesis deals with the detection and subsequent blood elements classification. Blood elements are described mainly in terms of optical features, that is used for digital image processing. Furthermore, available pre-processing theory, image analysis and methods of classifikation are discussed. Acquired knowledges are applied in MATLAB. Image Processing Toolbox is a pillar for that work within this programming environment. The program includes user interface, thus facilitating communication with the program. The program was tested on blood smear images and the results were evaluated.
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Fish excretion of ammonia under different oxygen environmental conditions
MÜLLEROVÁ, Lucie
The aim of this work was to find out and compare amount of ammonia nitrogen excreted by fish in different oxygen saturation of water. Experiments (preliminary, main and additional) were made with ornamental form of common carp (Cyprinus carpio f. Koi) at Laboratory of Aquatic Toxicology and Ichtyopathology RIFCH in Vodňany. The preliminary (orientation) experiment was made in three different dissolved oxygen concentrations (I. 0,65 - 2,66; II. 2,43 - 5,70; III. 6,40 - 6,50 mg.l-1 O2) and fish production of ammonia nitrogen after 2 hours was I. 24,61; II. 85,29; III. 70,19 mg N-NH4+ per 1 kg of fish weight. The fish were also exposed to different oxygen conditions (hypoxia0,76 - 1,46; normoxia 9,8 - 9,46 mg.l-1 O2) in the main experiment which was repeated six times. Detected production of ammonia nitrogen in recalculated to 1 kg fish weight was 25,01 +- 3,14 mg N-NH4+ by the fish in hypoxia and 32,56 +- 4,08 mg N-NH4+ by the fish in normoxia after two hours of experiment duration. Detected difference was statistically significant (P < 0,05). Haematology and biochemical blood examination of fish which were exposed to hypoxia proved statistically significant increasing number of leukocytes, value of haematocrit, concentration of ammonia and glucose was made in the end of experiment. The additional experiment was repeated five times, the fish were exposed to hypoxia there (0,99 - 1,21 mg.l-1 O2) and the control fish were kept in water with concentration of dissolved oxygen between 7,67 - 9,11 mg.l-1. Statistically significant differences in ammonia nitrogen production were already found out in these groups of fish after two hours of experiment duration and difference increased markedly after 4,5 hours (fish production of ammonia exposed to hypoxia 47,92 +- 4,91 mg.kg-1 and in the control fish 101,53 +- 8,31 mg.kg-1). Production of ammonia by the fish originally exposed to hypoxia almost aligned to production of control fish during 2,5 hours after an aeration had been added to the fish in hypoxia. Production of ammonia nitrogen was 93,38 +- 17,82 mg.kg-1 by the fish originally exposed to hypoxia and 110,13 +- 18,21 mg.kg-1 by the control fish. This difference was not statistically significant yet. The results of experiment proved that fish ability to excrete ammonia from body is closely connected with oxygen conditions in water environment.
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