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Role of endocytosis and endosomal acidification in TRAIL-induced apoptosis
Hradilová, Naďa ; Anděra, Ladislav (advisor) ; Kovář, Jan (referee)
TRAIL (TNF-related apoptosis inducing ligand) became known for its ability to selectively eliminate cancer cells. This ligand is a member of the TNF (tumor necrosis factor) ligands family and triggers extrinsic apoptotic pathway by binding of its death receptor 4 or 5 (DR4/5), and subsequent formation of death-inducing signalling complex (DISC). This signalling complex is required for successful transmission of apoptotic signal and activation of proximal caspases. However, regulation of the initial steps leading to activation of caspases is still not fully understood. Endocytosis of a TRAIL- DR4/5-DISC complex can be one of modulators of the initiation of extrinsic apoptotic pathway. Recent studies show controversial data documenting that endocytosis of TRAIL receptosomes can in cell type specific manner either positively or negatively influence TRAIL-induced apoptotic signalling. In this study, we focus on the analysis of a role of endocytosis and acidification of endosomal compartments during TRAIL-induced apoptosis in human colorectal cancer cell lines. Our results support the view that both clathrin-dependent endocytosis of TRAIL receptosome and endosomal acidification positively affect activation of caspases during the early stages of TRAIL-induced apoptosis. Inhibition of endocytosis or endosomal...
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Genetic factors of resistance to chemotherapy and clinical characteristics of breast tumors.
Kunická, Tereza ; Souček, Pavel (advisor) ; Kleibl, Zdeněk (referee)
Breast cancer is the most frequent malignancy in women population both in the Czech Republic and worldwide. Treatment of this disease involves surgical removal of the tumor, radiotherapy, chemotherapy and hormonal therapy. Recently, targeted biological treatment is also approached. Each patient reacts to the treatment individually and thus high variability in response is common. Multidrug resistance (MDR) presents one of the most important obstacles to successful chemotherapy. MDR is often associated with a decreased intracellular accumulation of anticancer drugs and an increased expression of ABC transporters such asABCC1 of our interest. The ABC family of membrane transport proteins includes the well-known mediators of resistance to anticancer drugs. In particular, ABCB1, ABCC1 and ABCG2 actively perform efflux of various types of drugs from cancer cells, thereby conferring resistance to those agents. The main aim of this study was to asses the genetic variability of the ABCC1 gene in 191 patients with breast cancer and to determinate the expression profile of ABCC1 in 30 patients from this cohort who were treated preoperatively. We evaluated relations between ABCC1 genotype, or phenotype and prognostic factors including the result of chemotherapy. Gene expression was measured in preamplificated...
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Interaction of polyomaviruses with proteasomal system of host cell
Verdánová, Martina ; Drda Morávková, Alena (advisor) ; Horák, Martin (referee)
Interaction of polyomaviruses with proteasomal system of host cells Abstract: Viral family Polyomaviridae includes besides model organisms - mouse polyomavirus and SV40 virus, also human pathogens, for example, BK virus. Polyomaviruses are small non- enveloped viruses with double-stranded DNA. Understanding of their life cycle is important for their use in gene therapy and immunotherapy as well as for prevention and treatment of complications caused by these viruses. This thesis is focused on early phases of MPyV and SV40 infection studying, mainly on delivery of viral genome to nucleus and role of proteasomal system in this stage of infection. It was found out that inhibition of proteasomes by specific inhibitor leads to increase of early non-structural protein LT expression, which was chosen as marker for viral entry to the nucleus and successful viral expression. Relative localization of proteasomes and VP1 protein of MPyV and SV40 was monitored and it showed 10% colocalization of mentioned structures. Further, it was found out that proteasomal inhibitor MG-132 negatively influences the replication of both viral and cellular DNA. Next aim of this diploma thesis was to prepare antigen - unique part of VP2 protein of BKV - for producing antibody. Expression vector with inserted fragment of unique part of...
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County Council and its Possibilities to Participate on Improving the Quality of Foster Care
DVOŘÁKOVÁ, Jana
The bachelor's work is is focused on parents, which desided to take a child into a foster care. Before this process, ``preparation for child acceptance into family{\crqq} preceded. Under our social conditions, the preparation is completely in authority of the county council. This institution makes a psychological adjudication of applicants, during which their character qualitites and reasons for child acceptance are assessed. On the basis of this adjudication, the applicant is declined or admit to peparatory course. The aim of the preparation course is to provide the potential parents with sufficient information essential for successful upbringing of the stepchild. The preparation has several parts and the decision of inclusion into applicants register is given at the end. Practical part includes two illustrative casustic interviews with foster families. They positively evaluate the activity of county council, but they also emphasize the necessity of permanent warning of wild range of potential problems and variety of their solutions.
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Preparation of expression constructs of phosducine binding partners.
Koláčková, Kateřina ; Obšil, Tomáš (advisor) ; Vaněk, Ondřej (referee)
Key words: SUG1, phosducin, G protein signaling, expression, pGEX-4T-1, pST39 Phosducin is an acidic protein found in cytosol and is involved in G protein signaling within rods of photoreceptors. Phosducin also influences stress-dependent hypertension. One of phosducin's important binding partners is SUG1, a 26S proteasome subunit, which belongs to a family of proteins that have an ATPase activity. This bachelor thesis takes part in a broader research, which tries to clarify physiological functions of the interaction of SUG1 with phosducin. Main goal of this study was to prepare constructs of SUG1 (human isoform 1) suitable for the recombinant protein production in an expression system E. coli BL21 (Rosetta). Gene that encodes SUG1 was inserted into two different plasmid DNAs that allow expression of SUG1 as two different fusion proteins in a prokaryotic expression system. Nucleotide sequences of these constructs were verified using sequencing. Next, expression tests revealed sufficient production of both recombinant fusion proteins. The main result of this bachelor thesis is the successful expression of human SUG1 in E. coli BL21 (Rosetta) cells. This result will enable the development of the purification protocol for the large scale expression of both recombinant fusion proteins as well as the...
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F. Kupka and his letters to V. Hynais
Brožová, Kristýna ; Wittlich, Petr (referee) ; Prahl, Roman (advisor)
This bachelor thesis, named F. Kupka and his letters to V. Hynais deals with mutual correspondence between František Kupka and Vojtěch Hynais. Their letters are currently placed in the National Gallery Archive in Prague. Dealing with this correspondence required translation from French to Czech and critical analysis of the text in comparison to already published studies. Many of the currently known information concerning Kupka has been confirmed while some new facts have also been uncovered. Although it may have seemed formal at first sight, the relationship between the two artists was truly warm. They shared not only their artistic points of view but also the joys and sorrows of their family lives. They often included magazine articles dealing with art and the current situation in the Czech and French art world into their letters. Kupka often mention his work, mainly book illustration, and expressed his longing for painting. His correspondence reassures us of his punctual preparation for each art project and also sometimes helps withs its dating. The reader is also confronted with Kupka's successes and failures with his search for a new artistic expression, although this can only be found in slight indications, not in connection to concrete paintings. The most important outcome of studying their...
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Portrait of Eleonora di Toledo in the Collection of the National Gallery in Prague
Táborská, Eva ; Zlatohlávek, Martin (advisor) ; Přibyl, Petr (referee)
Portrait of Eleonora di Toledo in the collection of National Gallery in Prague The main theme of my thesis is the study of one of the finest renaissance paintings in the collection of The National Gallery in Prague - Bronzino's Portrait of Eleonora di Toledo. Beside the main theme, the thesis also focuses on painting's context such as family relationship in which Eleonora had lived including a brief outline of general conditions in Florence around 1550. The thesis is set together on basis of secondary literature, which is noticed in the quotes. There is not a lot of information remained about Eleonora's own life, however we are able to develop a concrete idea about her personal life - a young lady from a distinguished Spanish family married the Duke of the one of the most significant Italian city states - Florence. She became a wife of Cosimo di Medici, she was a fine woman and a carefull mother, but also a lovely lady as shown in the portrait. When Agnolo Bronzino came up to produce Eleonora's portrait, he had been already treated as an accounted and acclaimed artist. Also Giorgio Vasari depicted his talent with honourable mention in his works. Particularly in Florence, he started his career of successful Medici portrait painter. As a turning point of his works, it is considered his painting of...
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Problematics of three-year old children adaptation to kindergartens
Benešová, Andrea ; Havlová, Jana (referee) ; Koťátková, Soňa (advisor)
The diploma thesis refers to a specific question of a kindergarten adaptation of three-year old children. The theoretical part covers the characteristics of a three-year old child from the point of view if his/her development level, necessaries of life, socialization, and psychic stability, and studies the coherence between the above-mentioned characteristics and the child's adaptation to a kindergarten. The practice-oriented part maps the most common problems with adaptation of three-year old children and compares the level of successful adaptation of three-year old children in homogenous and heterogeneous classes. Furthermore, this part studies also the interval of child's adaptation to his/her family, mainly the organizational and relationship changes. In the conclusion, the reasoning about possibilities of further use of this study for kindergarten teachers takes place.
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Preparation and study of human lymphocyte receptor LLT1
Bláha, Jan ; Vaněk, Ondřej (advisor) ; Konvalinka, Jan (referee)
Natural killer (NK) cells are an intensively studied part of immune system, possessing unique ability to recognize and induce death of tumor and virus-infected cells without prior antigen sensitization. Their function is regulated by a fine balance of signals induced by multiple activating and inhibitory cell surface receptors and their interaction with the ligands present on the target cell. Recent research in their C-type lectin-like receptors repertoire has shown that ligands of some of these previously orphan receptors lie within their own family, describing a lectin-lectin interaction. This is the case of human inhibitory receptor NKRP1 (gene KLRB1) and its ligand LLT1 (gene CLEC2D). Previous studies have shown that overproduction of LLT1 in cancer cells or lower production of NKRP1 in NK cells is connected to cancerous manifestations. This master's thesis shows a successful production of the extracellular part of LLT1 utilizing a mammalian expression system based on transient transfection of modified human embryonic kidney (HEK) cell lines. It was found that the five cystein residues contained within the lectin domain of LLT1 tend to cause misfolding and formation of aggregates. Stabilization of the domain was achieved by restoration of the sixth cystein residue at the evolutionary conserved...
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Cloning, detection and analysis of the gene cluster coding synthesis of the antibiotic Anthramycin
Poubová, Martina ; Černá, Milena (advisor)
he aim was to find , isolate and sequence a cluster of genes coding for the synthesis of anthramycin , antibiotics group pyrrolobenzodiazepines whose producer Streptomyces refuineus var . thermotolerans . Pyrrolo [2,1- c ] [1,4] benzodiazepines are a class of naturally occurring antibiotics anticancer effect. These are molecules that bind covalently to DNA molecules and this leads to blockage of transcription. Although anthramycin least affects the bone marrow, was not put into clinical practice as antitumor antibiotic because of the high cardiotoxicity. Better effects and good clinical features expected from a new synthetic analogues both in terms of antitumor antibiotics , and the production of antimicrobial compounds and antimalarial drugs . So far, successful antibiotic related families of antibiotics , lincosamide , was clindamycin antibacterial and antiparasitic effect. Knowledge of the genetic background of these substances allows modification simultaneously substances used , followed by a less costly production more efficient and safer substances. Using molecular biological methods were isolated part of the chromosomal DNA Producer Streptomyces refuineus var . thermotolerans containing the complete gene cluster anthramycin biosynthesis .
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