National Repository of Grey Literature 84 records found  beginprevious49 - 58nextend  jump to record: Search took 0.00 seconds. 
The expression of TP53 gene at the mRNA level in patients with myelodysplastic syndrome
Šeborová, Karolína ; Beličková, Monika (advisor) ; Španielová, Hana (referee)
Myelodysplastic syndrome (MDS) is a heterogeneous group of diseases characterized by ineffective hematopoiesis which is caused by damage of differentiation of pluripotent haematopoietic stem cells. TP53 gene mutations are identified approximately in 10% of MDS and represent a negative prognostic factor. Altered TP53 gene expression may have similar effect as the mutation. Mutations or deregulated expression of this gene have an impact on many cellular processes including apoptosis, DNA repair, cell growth and angiogenesis. In this work, the expression mRNA levels of genes involved in p53 signalling pathway were studied in CD34+ pluripotent haematopoietic cells from bone marrow of patients with low- risk MDS. MDS patients showed increased expression of genes involved in apoptosis induction, regulation of cell cycle and DNA repair (BAX, BBC3, CCNE1, CDC25A, CDKN1A, FAS, GADD45A) as compared to healthy subjects. The patients with TP53 mutation had decreased expression of apoptotic genes (BAX, PIDD, TRAF2) and increased gene expression of apoptotic inhibitor (BCL2A1), indicating a reduced activity of apoptotic pathways and that way the pathological cell clone may gain a growth advantage. Deregulation of 21 genes (BAX, BBC3, EGR1, KAT2B, MDM2 etc.) was observed in patients with del (5q) compared to...
Utilization of mouse polyomavirus derived virus-like particles for cargo delivery into cells
Polidarová, Markéta ; Španielová, Hana (advisor) ; Šmahel, Michal (referee)
and key words Mouse polyomavirus-derived virus-like particles composed from major capsid protein VP1 (MPyV VP1-VLPs) are interesting structures for use as a delivery system of various cargos into cells. VP1 protein self-assembles into icosahedral particles of 45 nm in diameter that are hollow highly regular nanoparticles. In this work, model small molecule cargo, Cyclodextrin-Based Bimodal Fluorescence/MRI Contrast Agent, was encapsidated into MPyV VP1-VLPs. The cargo was stably associated with VLPs and was delivered into mammalian cells using these VLPs. To prevent VLPs entrapment in endolysosomal compartments and increase the potential of VLPs applications, MPyV VP1 protein was modified by insertion of histidine-tag (6 histidine long sequence surrounded by glycine and serine) sequences into VP1 surface loop DE, because histidine modification of synthetic systems had enhancing effect on endosome escape and cargo delivery. With the use of in Bac-to-Bac® baculovirus expression system His-VP1 protein was expressed in insect cells and a variety of VP1-assemblies was obtained: long tubules and small 20nm VLPs formed from VP1 with 4 histidine-tags in DE loop, and novel VP1 nanostructure, which we named nano-jumpers, formed from VP1 with 2 histidine-tags. Nonetheless the endosome escape properties of...
The longevity in mole rat.
Jelínková, Alena ; Schierová, Michaela (advisor) ; Španielová, Hana (referee)
Naked mole rat and blind mole rat are useful model organisms for human age-associated diaseases studies. Unlike human, their long lifespan is not accompanied by physical health impairment. In both species, the genes involved in aging process or carcinogenesis are under positive selection or their regulation differs from the regulatory pattern known in other rodents or human. Some genes are present in higher number of copies, missing or entirely new and not observed in other organisms. In naked mole rat, the degenerative development is reduced by elevated level of proteins which prevent amyloid β aggregation and contribute to oxidative damage tolerance. Their healthy aging is also caused by effective elimination of damaged proteins, natural caloric restriction or angiogenesis enhancement. High level of α-2-macroglobulin in blood, which is able to inhibit signal pathways required for tumor growth and malignancy, as well as the early contact inhibition repress tumorigenesis in naked mole rat. Many different mechanisms are involved in prolonged lifespan in both naked and blind mole rat species. The aim of this thesis is to present the most important genome and proteome differences contributing to their long lifespan. Key words: naked mole rat, blind mole rat, tumorigenesis resistance, senescence,...
Optimization of methods for analysis of early steps of mouse polymavirus life cycle
Soukup, Jakub ; Španielová, Hana (advisor) ; Němečková, Šárka (referee)
Mouse polyomavirus is a type species of Polyomaviridae family and serves as model for studying viral infection of human pathogenic polyomaviruses. Minor proteins of viral capsid have been found to be necessary for effective initiation of infection. In order to study their role in the early steps of infection we utilized the novel Cre-LoxP system for production of the viral mutant lacking both minor proteins. Virus produced this way was compared with virus produced by standard method and we found that both systems facilitate production of mutant virus with the comparable quality and quantity. The mutant virus contained reduced amount of viral DNA and formed virions with impaired stability. For further studies of intracellular virion trafficking we prepared virions with genomes modified by thymidine analogues 5- bromo-2'-deoxyuridine (BrdU) and 5-Ethynyl-2'-deoxyuridine (EdU) and optimized the methods for analogue detection. The viral genome become accessible for detection 4 hours post infection. For ultramicroscopic analysis of translocation of virus to the nucleus we used freeze substitution. All this methods will be utilized for detailed study of distinct steps in viral infection. Key words: Mouse polyomavirus, minor proteins,...
Directed evolution of mouse polyomavirus
Váňová, Jana ; Španielová, Hana (advisor) ; Mašek, Tomáš (referee)
The method of directed evolution represents a new approach to generate proteins with new or altered properties. The principle of directed evolution is random mutagenesis of the coding sequence for a protein of our interest followed by selection of generated mutants for the desired property. The aim of this pilot study was to investigate the possibility of utilization of directed evolution for alteration of mouse polyomavirus original tropism and virus retargeting to a model prostate cancer cell line. To generate randomly mutated gene encoding the major capsid protein of mouse polyomavirus, which is responsible for the interaction of the virus with cellular receptor for viral cell entry, error-prone PCR and DNA shuffling methods were used. Production of viruses composed of mutant major capsid protein was ensured by Cre/loxP site-specific recombination. The thesis also dealt with the design and characterization of the system for viral mutant selection. It was found that the prostate cancer cell lines markedly vary in their ability to bind and internalize particles derived from mouse polyomavirus. This knowledge can be used for the preparation of virus-like particles for prostate cancer diagnostics in the future. The study demonstrated that the method of directed evolution can be used for production...
Reporter expression system for study of silencing of provirus integrated inside transcriptionally active gene
Slavková, Martina ; Hejnar, Jiří (advisor) ; Španielová, Hana (referee)
Retroviral vectors are used as mighty tools for an introduction of recombinant genes into the recipient genome in gene therapy trials. In the vector design, great emphasis is put on safety and efficiency. In spite of a great progress in retroviral vector design with the purpose to stabilize its expression, e.g. introduction of protective elements into the viral regulatory sequences, the current approaches are still not sufficiently effective and the majority of proviruses is transcriptionally silenced. The understanding of the silencing mechanism is of special importance to the optimization of the vector design and handling. In this master thesis, I have designed and constructed an expression system for study of the mechanism involved in the silencing of retroviruses integrated inside gene bodies. This artificial system will be utilized for testing of hypothesis that retroviruses integrated into gene bodies are silenced by DNMT-dependent mechanism and this process is triggered by transcriptional read-through of the provirus from nearby host promoter. I have obtained preliminary results suggesting the validity of the suggested hypothesis; however the verification of general validity of this hypothesis for various retroviruses and elements will be a matter of further studies in our laboratory. Powered by...
Genotypic analysis of human cytomegalovirus in the patients after allogeneic haematopoietic stem cell transplatation.
Javornická, Tereza ; Hubáček, Petr (advisor) ; Španielová, Hana (referee)
In patients after allogeneic haematopoietic stem cell transplantation (HSCT) is a human cytomegalovirus (CMV) one of the most important viral pathogens. Its detailed characteristic could provide information about the impact of each CMV genotype on overall survival of the patient, and some serious complications, such as graft versus host disease (GvHD). This thesis deals with retrospective genetic analysis of samples from 1877 patients transplanted at the Clinic of Pediatric Hematology and Oncology, University Hospital Motol and the Institute of Hematology and Blood Transfusion since 2002. DNA from biological samples (especially whole blood) was isolated kit Qiagen DNA Blood Mini or Qiagen DNA Mini and samples were prospectively detected presence of CMV DNA. Samples were subsequently stored at -20 řC. Genotyping was performed using real-time PCR technologies to the genes of 2 structural proteins glycoprotein B, glycoprotein H and using sequence specific primers and probes. In 1343 samples (71.6%) from 390 patients there was only one strain of CMV; in 256 (13.6%) samples from 113 patients have detected mixed infection caused by two or more strains of CMV. The most common genotype demonstrated in "single" infection was in pediatric and adult patients gB1/gH2 detected in 118 (28.4%) patients. Most...
Stratification risk of disease progression in patients with abnormal cervical cytologic finding by means of molecular genetic analysis of selected biological factors
Gomolčáková, Barbora ; Kašpírková, Jana (advisor) ; Španielová, Hana (referee)
The aim of this thesis was to track the impact of selected herpesviruses, polyomaviruses, Chlamydia trachomatis and methylation of tumor supressor genes at the development and progression of high grade- lesion in HPV - positive patients by means of molecular-genetic techniques. Confirmation of these markers presence in women with severe lesions of cervix would help to raise necessary specificity of molecular genetics HPV testing and recommend it as a primary screening test for cervical carcinoma prevention. HPV testing could thus replace currently prevailing cytology which has relatively low sensitivity and therefore the number of false negative results. The analyzed samples consisted of cytological cervical smears of 51 HPV positive women, with histologically confirmed presence of severe lesions, collected in liquid medium. Samplings from 51 women without infection were used as a control. The possible effect on disease progress was confirmed only in the case of gene promoters' methylation whose presence was detected in up to 26 patients. It is, however, very unlikely that cancer would develop in all these women. This marker could thus help to stratify patients at risk but only to some extent. Although the individual effect of remaining markers has not been established in the carcinogenesis of cervical...
The application of viral nanoparticles in biomedicine
Kříž, Kristian ; Španielová, Hana (advisor) ; Beranová, Jana (referee)
This work compiles studies that lead to new applications of viral nanoparticles with an emphasis on utilizations in biomedicine. Firstly the work follows the characteristics of viral nanoparticles and some general aspects of nanoparticles. Further described herein are exemplary studies that are using viral nanoparticles as a tool for the treatment of cancer, viral, bacterial or other diseases. This review also includes examples of researches leading to the usage of viral nanoparticles for diagnostic purposes. Lastly, studies in which viral nanoparticles served as nanoextractors, nanoreactors or nanotemplates for the synthesis of metal nanoparticles and galvanic cells are mentioned here too.

National Repository of Grey Literature : 84 records found   beginprevious49 - 58nextend  jump to record:
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