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Mutační status a jiné chromozomální změny u chronické lymfatické leukémie
ŠLOUFOVÁ, Martina
This thesis is focused on mutation status and other chromosomal changes in chronic lymphocytic leukemia. CLL is a lymphoproliferative illness with a low malignancy based on clonal proliferacy of malignatly transformed B lymphocytes. Although this is the most frequent malignant illness in western population, the cause leading to its origin has not been yet clearly given. Identifying prognostic markers has a significant meaning in identifying the illness prognosis, the choice of treatment strategy, and it influences general survival of patients. The aim of this thesis is to describe the frequency of the most usual mutations in patients suffering from CLL. All the results obtained from CLL patients were acquired from the Hospital in České Budějovice from 2016 to 2018.
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Detekce a kvantifikace inhibitorů proteáz v klíštěti \kur{Ixodes ricinus} pomocí monoklonálních protilátek
VANÍČKOVÁ, Martina
Inhibitors of proteases in tick saliva play an important role during tick feeding. Tick saliva contains a wide range of bioactive components which are able to modulate host imunity. Therefore, ticks are able to feed for a long time and transfer tick-borne diseases pathogens. The risk of transfer can be significantly reduced by deactivation of theese protease inhibitors. In this study I made monoclonal antibodies for detection and quantification of two serine protease inhibitors in tick saliva and other tick-body parts.
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Radiolabelled antibodies - the perspective for diagnosis and therapy
Mejtská, Jana ; Bárta, Pavel (advisor) ; Ramos Mandíková, Jana (referee)
1 CHARLES UNIVERSITY FAKULTY OF PHARMACY IN HRADEC KRÁLOVÉ DEPARTMENT OF BIOPHYSICS AND PHYSICAL CHEMISTRY DISSERTATION THESIS RADIOLABELED ANTIBODIES - THE PERSPECTIVE FOR DIAGNOSIS AND THERAPY Supervizor: Mgr. PAVEL BÁRTA, Ph.D. HRADEC KRÁLOVÉ, 2017 Bc. JANA MEJTSKÁ 2 ABSTRACT EN Different types of tissues have a characteristic cell morphology. Each cell has typical molecules on its surface, which may be either of physiological or pathological type. The presence of these surface structures can be interesting for possible modulation of specific cell populations from neighboring cells. Utilization of this property is then essential particularly in a case of tumor cells. Targeting on tumor specific cell structures involves the use of receptor specific peptides or monoclonal antibodies. The discovery of the preparation of monoclonal antibodies has opened a new chapter in the treatment and diagnosis not only tumor diseases. The advantage of monoclonal antibodies is their specificity and also high affinity to the type of the target cell structures. This study is focused on the summary of monoclonal antibodies which are currently being applied on the treatment or diagnosis of a particular cancer. Furthermore, this work also includes antibodies which are under development for intended medical applications with...
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New approaches in vaccination against HIV
Dobiášová, Julie ; Drda Morávková, Alena (advisor) ; Strachotová, Dita (referee)
A B S T R A C T Most current vaccines are based on using whole-inactivated viruses. After creating the immune response and immune memory is organism able to cope with infection create by patogens. In the case of HIV, however, fail to produce the vaccine, which would have been able vaccinated individual from subsequent infections protect. Virus HIV attacks CD4+ cells and destroys the immune system. Rate of his replication is high and virus HIV is resistant to existed antivirotics. And he is resistant before cells, which conveying the immune response. Moreover, the virus persists in cell in the form proviral DNA. For a successful vaccine against HIV is developed a lot of new vaccines and vaccination procedures. One way is the using recombinat viral glycoproteins, which are incorporated into the membrane of virus HIV, which should produce in the vaccinated organism production of neutralizing antibodies. Some modern models of vaccines strategies don't target the virus itself, but they target the restriction of HIV infection by destroying infected cells via apoptosis, or cytokine secretion. Using plasmid DNA cobination with recombinant vectors appear as the most perepective opportinity to develop HIV vaccine. Unfortunately, traditional models or new models of vaccine against HIV are failing to provide a...
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Diversity of methods used for characterization of molluscan hemocytes
Jindrová, Zuzana ; Skála, Vladimír (referee) ; Horák, Petr (advisor)
Hemocytes are the main immune cells of invertebrates; therefore they can be found in molluscs, too. They differ both in morphology and function. The two generally accep- ted morphological types, granulocytes and hyalinocytes, vary in the level of phagocy- tosis and encapsulation, production of reactive oxygen species and nitrogen oxide, and presence of some enzymes. There is an array of methods by means of which hemocy- tes can be characterized. Microscopy serves particularly for study of morphology. An- tigens localized on the surface can be determined by monoclonal antibodies or lectin probes. Hemocytes can be divided on the basis of cell size and granularity using gra- dient centrifugation or flow cytometry. Production of nitrogen oxide and reactive oxy- gen species is monitored by adding appropriate substrate which changes its proper- ties after reaction with the radical. It may become fluorescent, change absorbance of the solution or form a visible precipitate. Another possibility is the use of chemilu- miniscence. The objective of hemocyte research is to explain mollusc-pathogen inter- action. 1
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Epididymal maturation – a crucial step in the post-testicular sperm development
Postlerová, Pavla ; Pohlová, Alžběta ; Zigo, Michal ; Jonáková, Věra
Mammalian spermatozoa after their development in testis undergo the post-testicular maturation in epididymis where acquire their fertilization ability and competence of movement. The epididymis is tissue with very active fluid-absorbing and fluid-secreting activity. Epididymal fluid contains ions and small molecules, proteins, glycoproteins and enzymes. The surface of spermatozoa is exposed directly to the epididymal fluid, and the sperm plasma membrane is significantly changed. Some testicular proteins are altered, masked, or replaced by new proteins/glycoproteins of epididymal origin. Several proteins produced by epididymis have been described in various mammalian species and shown to be associated with spermatozoa suggesting a role in the sperm maturation and/or sperm-egg binding and fusion. We isolated proteins from fluid, tissue and sperm of boar epididymis, and separated them by chromatographic and electrophoretic methods. We searched for known proteins using panel of antibodies and tested proteins of epididymal fluid for binding abilities. In the epididymis, we found proteins described as proteins of seminal plasma and associated with the sperm surface, such as spermadhesins, beta-microseminoprotein and acrosin inhibitor. These proteins were detected in epididymal sperm, fluid and tissue. We showed that some epididymal proteins may bind the spermatozoa and change the binding sites on the sperm surface. We determined and identified some proteins from boar epididymal fluid with affinity to heparin, hyaluronan and zona pellucida glycoproteins. These phenomena indicate that epididymal fluid proteins bind to the sperm surface during epididymal maturation and might subsequently play role in the sperm capacitation or sperm-zona pellucida binding.
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Estrogen receptor beta (ERβ) in testicular cells and sperm
Dostálová, Pavla ; Žatecká, Eva ; Děd, Lukáš ; Dorosh, Andriy ; Postlerová, Pavla ; Jonáková, Věra ; Dvořáková-Hortová, Kateřina ; Pěknicová, Jana
Estrogen is a steroid hormone that plays an important role during sperm development in the male and female reproductive tract. Estrogen signalling is a complex process that depends on cell milieu and presence of receptors. Thanks to the steroid nature of estrogens, they can pass through the plasmatic membrane and bind to the intracellular estrogen receptors (ERs). Within the cell, there are several pools of ERs. One of them is localized to the cell nucleus and their activation leads to direct or indirect binding to DNA and ultimately to alternation in gene expression (genomic pathway). Other pools of ERs are associated with plasma membrane or are located in cytosol. Activation of membrane associated ERs leads to rapid non-genomic responses. Nowadays, two classical estrogen receptors are known – ERα and ERβ. Since ERβ is a predominant variant in testes, we focused our study on expression of ERβ variants in murine testes and sperm. We detected two variants of ERβ at mRNA level in both, testes and sperm. These variants differ in 54 nucleotids within the ligand binding domain and this variability results in different affinity to estrogen. We analyzed individual testicular cell types (spermatogonia, spermatocytes, spermatids, Sertoli cells) by RT-qPCR. Our results suggest that both ERβ variants are coexpressed in the same cell type and may therefore interact together. This may have consequences in mediating of estrogen signalling. Moreover, ERβ is expressed more in the later stages of spermatogenesis suggesting the role of ERβ in these stages or alternatively in spermatozoa alone. At the protein level, we detected ERβ in nuclear, membrane and cytosolic fraction prepared from testicular tissue suggesting the involvement of both, genomic and non-genomic, pathways of estrogen signaling in testes. In sperm, anti-ERβ antibodies localized ERβ in acrosome region and tail which is in accordance with the known role of estrogen on capacitation, acrosome reaction and motility.
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Preparation of monoclonal antibodies against Borrelia
ŠVEJDOVÁ, Veronika
In my bachelor thesis I deal with preparation of monoclonal antibodies against antigens of Borrelia. In theoretical section I describe genus Borrelia and antibodies. The introduction deals with the history of discovery of the spirochete that causes Lyme disease, vectors of this disease and classification of Borreliae according to geographical occurrence. Taxonomic categorization and description of the bacterium followers. It is focused on flagellum, arrangement of genetic material and on external surface antigens important for this bacterium. In the next section I describe Borrelia vector - host interactions and the development of immune response during the disease. Dermal symptoms of the Lyme disease are described in detail. A few words about relaxing fever are added. This chapter ends with description of laboratory diagnostics of Borrelia burgdorferi. Next, I focused on the characterization of antibodies and on preparation of monoclonal antibodies by hybridoma technology. I briefly describe ways of their utilization. Practical part of Bachelor thesis was carried out in the laboratories of Faculty of Science. Here, with the help of hybridoma technology, I fused myeloma cells with splenocytes of immunized mice producing hybridomas. These hybridomas I cultivated in medium, which I subsequently tested for occurrence of antibodies by ELISA. Hybridomas having high productivity were cloned, frozen and stored in liquid nitrogen. Finally I purified monoclonal antibodies, concentrated them and tested their titre.
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