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Signaling pathways and genes regulating oocyte maturation in pig
Blaha, Milan
The gonadotropin-induced resumption of meiosis and cumulus expansion in preovulatory follicles is preceded by expression of epidermal growth factor (EGF)-like factors, amphiregulin (AREG) and epiregulin (EREG), in mural granulosa and cumulus cells. In vitro, the EGF-like peptides are also produced in cumulus cells upon stimulation by FSH. Both FSH and the EGF-like peptides stimulate resumption of meiosis and cumulus expansion in vitro via activation of a broad signaling network in cumulus cells. To define signaling pathways that drive FSH- and AREG-induced cumulus expansion and meiotic resumption, in vitro cultured pig cumulus-oocyte complexes (COCs) were treated with specific protein kinase inhibitors. The results document that FSH-stimulated, but not the AREG-stimulated resumption of meiosis, depends on the PKA and MAPK14 activities; both modes of stimulation require activation of EGFR and MAPK3/1. To characterize the effects of FSH and EGF-like peptides on gene expression in cumulus cells, transcriptomes of cumulus cells were analysed using microarray approach. Both FSH and AREG+EREG increased the expression of genes associated with regulation of cell proliferation, blood coagulation and extracellular matrix remodeling. In contrast to AREG+EREG, FSH also increased the expression of genes coding...
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Methods of molecular and cell biology in the frame of secondary school education
Blaha, Milan ; Ehler, Edvard (advisor) ; Pavlasová, Lenka (referee)
Teaching of cell and molecular biology is often restricted by financial and material limitations as well as a lack of expertise. These problems may be overcome by cooperation with scientific institutions and universities. Some scientists teach select courses under the auspices of high school. Others allow students to fulfill internships at their workplaces. The first aim of this thesis is to describe the characteristics of summer school, wherein the participants are integral for providing feedback on what matters most to them. The second goal of the study is to perform four molecular biology lab lessons in the advanced placement genetics course. The results of a qualitative research conducted among summer school participants have shown the benefits present due to contact that occurs between people of the same genuine interest. Student evaluation of molecular biology lessons performed in the advanced placement course revealed that the lab lessons are feasible and positively accepted by students. The thesis also highlights the importance of cooperation between high school and academia. Key words: molecular biology, DNA, secondary schools, gifted students
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Signaling pathways and genes regulating gonadotropin-induced maturation of porcine oocytes and cumulus expansion
Blaha, Milan ; Němcová, Lucie (advisor) ; Král, Jiří (referee)
In vitro, meotic maturation of porcine oocytes and cumulus expansion are induced by FSH and EGF-like peptides AREG and EREG. FSH and EGF-like peptides induce expression of cumulus expansion-related genes (HAS2, PTGS2 and TNFAIP6). To define signaling pathways that control FSH- and AREG-induced cumulus expansion, porcine cumulus-oocyte complexes were treated with specific protein kinase inhibitors. Inhibitors of MAPK3/1, MAPK14 and ERBB1 significantly reduced both FSH- and AREG-induced expression of HAS2, PTGS2 and TNFAIP6. These inhibitors decreased FSH/LH-induced expression of AREG and EREG in mural granulosa cells. Surprisingly, inhibitor of PKA had no effect on AREG expression in cumulus-oocyte complexes but the inhibitor decreased expression of TNFAIP6 induced by AREG. Inhibitor of PI3K increased expression levels of AREG and PTGS2 but EREG, HAS2 and TNFAIP6 were reduced. Expression levels of the cumulus expansion-related genes were not affected by an analog of cGMP (8-CPT-cGMP). However, 8-CPT-cGMP blocked spontaneous in vitro meiotic maturation of porcine oocytes and its effect was abolished by FSH. Key words: cumulus expansion, cumulus expansion-related genes, meotic maturation, FSH, amphiregulin, cGMP
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The Quality and Safety of Transfusion Products - a Study on Critical Aspects of Quality and Important Factors in Quality Control
Zimová, Renata ; Bláha, Milan (advisor) ; Gašová, Zdenka (referee) ; Turek, Petr (referee)
The quality and safety of transfusion products - a study on critical aspects of quality and important factors in quality control Products made from blood (transfusion products and blood derivatives) represent a special group of therapeutic preparations that have specific properties (danger of allergic reactions, transfer of infections etc.). The aim of this study was to show critical points of the process of producing safe, first-rate transfusion preparations at the level reflecting the progress in medicine and technology. Secondary aim was to analyze whether inspections of the State Institute for Drug Control, strict requirements and implementation of EU legislative have influence on the quality of work in blood establishments (BEs) and whether there are some relations between selected BEs with similar characteristics (the size and type of facility) and quality of work as this is presently a heated topic in discussions about restructuralization of transfusion service. The author analyzed her own experience gained during 129 inspections from the view of Good Manufacturing Practice (GMP) in BEs in the Czech Republic and abroad. The survey of results The most frequent and most important shortcomings (key and critical points) in production of transfusion products are parameters described in the chapter...
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The Effect of EGF - like Peptides on Cumulus-Oocyte Complexes
Blaha, Milan ; Krylov, Vladimír (referee) ; Němcová, Lucie (advisor)
In mammals, the ovarian cycle is controlled by follicle-stimulating hormone (FSH) and luteinizing hormone (LH), secreted by the anterior lobe of the pituitary gland. FSH is assumed to be essential for follicle growth. LH is required for the terminal phases of follicle growth, estrogen synthesis, ovulation, development of the corpus luteum and progesterone synthesis. Although cumulus cells and oocytes lack LH-receptors, LH causes cumulus expansion and meiosis resumption. The effect of LH is mediated via EGF-like peptides - amphiregulin (AREG), epiregulin (EREG) and betacellulin (BTC). The precursors of these peptides are present in the mural granulosa cells and EGF-like peptides are released by activation of metalloproteases in response to LH-stimulus. The aim of this thesis is to present EGF-like peptides in the wide context of the molecular mechanisms regulating the late phases of folliculogenesis and meiotic maturation of oocytes.
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Revision of the electronic diabetic file at Clinic of Diabetology IKEM
Fialová, Lenka ; Blaha, Milan (referee) ; Provazník, Ivo (advisor)
Electronic diabetic record (EDR) is an electronic medical record used in Diabetology Clinics, Instittute for Clinical and Experimental Medicine (IKEM), Prague. EDR is complex record of patient statistical and clinical data. It´s complexity and large time demands on filling in all items make EDR not user friendly. Thus, benefits of electronic medical record are not fully utilized. The paper describes solution that leads to simpler user interface as component of outpatient message allowing easy entering data. The new interface has been designed and created in the form of program module. There are described other procedures to implement module into IKEM information system Zlatokop. At the end, there are discussed the advantages and shortcomings of this revised vision and current status of project of EDR. The first part of the work is about the basic concepts and problems regarding EDR. It is about e-health, specially about the electronic health record, databases, specially about Caché, which is used in Zlatokop, about diabetes mellitus and about user experience.
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Detection and analysis of eye movement
Křístek, Jakub ; Blaha, Milan (referee) ; Provazník, Ivo (advisor)
This thesis deals with the movements of eyes and it describes the matters related to its measurement and data processing. Theoretic part is focused on eye‘s anatomy and electrophysiology, types of eye’s muscles and their movements. Even more methods, not just used electrooculography, are described thoroughly. In the second part of this thesis are designed two experiments and the measurement is realized on a group of ten tested subjects. Acquired signals are saved and processed in Matlab software, statistically analyzed and the results are interpreted.
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