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GENE EXPRESSION AND IMMUNOLOGICAL RESPONSE IN MICE EXPOSED TO ZnO NANOPARTICLES
Rössner ml., Pavel ; Vrbová, Kristýna ; Strapáčová, S. ; Rössnerová, Andrea ; Ambrož, Antonín ; Brzicová, Táňa ; Líbalová, Helena ; Javorková, Eliška ; Zajícová, Alena ; Holáň, Vladimír ; Kulich, P. ; Večeřa, Zbyněk ; Mikuška, Pavel ; Coufalík, Pavel ; Křůmal, Kamil ; Čapka, Lukáš ; Dočekal, Bohumil ; Šerý, Omar ; Machala, M. ; Topinka, Jan
We analyzed gene expression changes in the lungs and the immunological response in splenocytes of mice exposed by inhalation of ZnO nanoparticles - NP. Adult female ICR mice were treated for three days and three months, respectively. Analysis of differential expression in genes involved in oxidative stress was conducted using quantitative RT-PCR. The potential immunotoxic and immunomodulatory effects of ZnO NP were analyzed by phenotyping and cytokine production by splenocytes after three months exposure. Three days exposure resulted in down-regulation of GCLC, GSR, HMOX-1, NQO-1, NF-kB2, PTGS2 and TXNRD1 mRNA expression, three months exposure increased the expression of these genes. Three months exposure caused a significant decrease in the percentage of granulocytes in the spleen cells, and affected the production of IL-10 and IL-6 by lipopolysaccharide-stimulated leukocytes. In summary, our study revealed changes in the expression of genes involved in the oxidative stress response following acute ZnO NP exposure. Subchronic ZnO NP exposure induced immunomodulatory effects in the spleen.
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Intensive care of patient with metformin intoxication
Zajícová, Alena ; Povolná, Pavla (advisor) ; Hocková, Jana (referee)
Metformin is the first-line oral antidiabetic drug for patients with type 2 diabetes mellitus. As the rate of new diagnosed cases of diabetes increases year-on-year, the number of patients who are prescribed metformin rises as well. In the Czech Republic only, 858,010 patients were treated for diabetes in 2015. 786,586 of them were treated for the type 2 diabetes mellitus. Metformin intoxication, called also MALA (metformin- associated lactic acidosis) after its main symptom, is a rare complication (≤ 10 cases per 100,000 patients per year) in the metformin treatment, but the mortality is high (up to 50%). Metformin-induced lactic acidosis develops when the contraindications of metformin therapy are not respected or in the event of an acute disease linked with dehydration and hypoxia. Patients with MALA are hospitalized at the int ernal medicine intensive care unit or the department of anesthesiology and critical care with multiple organ failure. This thesis aims at determining whether procedural nursing care standards, implemented in the form of a nursing process, are actively used in practice on intensive care beds. Methodology: The empirical part is compiled in the form of a qualitative research, as a case study of a female patient suffering from metformin intoxication, caused by a suicide...
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Effect of growth factors and bioactive substances on mesenchymal stem cell differentiation.
Blahnová, Veronika ; Daňková, Jana (advisor) ; Zajícová, Alena (referee)
Mesenchymal stem cells are a population of multipotent cells, which have the ability to restore damaged tissues derived from mesoderm. Under the influence of wide range of growth factors, hormones and other bioactive molecules they can differentiate for example into chondrocytes, osteocytes, adipocytes, myocytes or insulin producing cells. The differentiation is induced even by contacts with neighboring cells or with extracellular matrix. Tissue engineering often uses especially growth factors. Growth factors act through specific surface receptors, which mediate cell-to-cell and cell-to-extracelular matrix communication and influence many cellular processes such as growth, proliferation, differentiation and others. Due to their specific impact growth factors are used individually or in combinations in tissue engineering applications. Substances stimulating cells to proliferate and differentiate can be added right to the culture medium, when cultivated in vitro, or can be loaded into a scaffold onto which cells are later seeded. The determination of exact growth factors combinations and concentrations influence on MSCs differentiation would enable more efficient use of MSCs in regenerative medicine. The aim of this thesis is to summarise present knowledge about the effect of growth factors and other...
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Immunomodulatory properties of vitamin D3
Urbanová, Anna ; Stříž, Ilja (advisor) ; Zajícová, Alena (referee)
1 Abstract Vitamin D3 is important for keeping the right concetration of Ca2+ in plasma. Therefore it is essential for proper bone growth and development. Nevertheless, vitamin D3 has also a number of immunomodulating effects. Our thesis has been targeted on evaluation and comparison of vitamin D3 influence on expression of chosen surface markers (CD14, CD54, HLA-DR, CD16, CD36 and CD163) with THP-1 cells and monocytes gained from human peripheral blood. Other aims have been analysing the vitamin D3 influence on longevity of THP-1 cells and measuring the soluble CD14 and IL-8 production with THP-1 cells under the vitamin D3 influence. The cells have been stimulated with five different concentrations of vitamin D3 for the time 24, 48 and 72 hours. Higher used concetrations of vitamin D3, i.e. 100 nM and 1000 nM have increased the expression of CD14 with THP-1 cells in the time 48 and 72 hours of the stimulation time. With the monocytes from peripheral human blood the increase of the CD14 expression hasn't been remarkable from the physiological point of view. Together with the vitamin D3 concentration increase the sCD14 production with THP1 cells was considerably higher. The sCD14 was the highest in the time 72 hours after the stimulation with the highest used vitamin D3 concetration. The IL-8 quantity with...
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Impacts of chemotherapy on imunoregulatory gene expression in the tumor microenvironment
Paračková, Zuzana ; Reiniš, Milan (advisor) ; Zajícová, Alena (referee)
Tumor microenvironment is an area, where the local immunosuppressive effects dominate and prevents the immune system to perform its physiological functions. The cells infiltrating the microenvironment have an important function among many cell types since they produce a large quantity of factors suppressing the immune response. In our work, we monitored the immune changes in the microenvironment during tumor growth and chemotherapy. For these purposes, we utilized the methods for analysis of the proportion and phenotype of the distinct populations of immunocytes and for analysis of the total level of expressions of selected genes associated with immunosuppression or with distinct populations of immunocytes. The aim of our work was to discover, using two types of mouse tumors (TRAMP-C2 and TC-1/A9), how 5-azacytidine (5AC), a cytostatic drug with epigenetic activity, affects the proportion of leukocytes infiltrating the tumor microenvironment and, further, whether these changes are accompanied by decreased expression of immunosuppressing genes. In addition, we have also focused on the changes of relative expression of genes encoding markers of lymphoid lines and, on other immunoregulating genes, encoding IL-6, IL-10, IL-12, IL-4 and IFNγ cytokines, in the microenvironment of these tumors....
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Modulation of mesenchymal stem cell properties and their use in the regulation of transplantation immunity
Peřinová, Lucie ; Krulová, Magdaléna (advisor) ; Zajícová, Alena (referee)
Mesenchymal stem cells (MSCs) represent a heterogeneous population of stromal cells with a pluripotent differentiation potential. They can be isolated from multiple tissues of mesodermal origin, such as bone marrow, adipose tissue, umbilical cord blood and peripheral blood and afterwards externally expanded according their adherence to the plastic surfaces. These cells show remarkable immunomodulatory properties, suppressing T-, B- and NK-cell functions, and also modulating dendritic cell activities and influencing immune responses during tissue repair and recovery. MSCs have been shown to possess ability to migrate to sites of inflammation and tissue injury. All these properties make MSCs a promising tool for clinical application. Our primary goal was to identify processes that may influence immunoregulatory effects of MSCs. In order to promote immunossupressive qualities of MSCs we established the scheme comprising MSCs precultivated with various cytokines and Toll-like receptors (TLR) ligands in vitro, with the final aim to improve the therapeutic effect of MSCs on wound healing in vivo. We studied modulation of MSCs properties and consequently the effect of influenced MSCs on cells of the immune system. The immunosuppression is mainly mediated through secreted factors that MSCs produced after...
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Regulation of the cytokine IL-33 production and its biological effects.
Kardošová, Miroslava ; Stříž, Ilja (advisor) ; Zajícová, Alena (referee)
IL-33 is a dual function protein that may function as both a proinflammatory cytokine and an intracellular nuclear factor. In a role of cytokine IL-33 signals via receptor ST2 and induces T helper type 2-associated cytokines in its target cells including mast cells, basophils, eosinophils and natural killer cells. Additionally, it acts as chromatin-associated nuclear factor with transcriptional regulator properties affecting expression of some proinflammatory cytokines. Regulation of this processes is poorly understood, mechanisms underlying synthesis, processing and secretion of IL-33 also remain to be fully explored. The aim of our study was to examine mechanisms probably involved in regulation of IL-33 production and its secretion outside the cell. First, we investigated possibility that IL-33 secretion is affected by stimulation with cytokines TNFα, IFN γ, IL-1β, IL-13, IL-33, TGF-β and IL-10 or stimulation with LPS isolated from E. coli. Next we investigated hypothesis that IL-33 is released from cells during cell damage or necrosis and serve as "alarmin". Necrosis was induced in LPS-stimulated cells by freeze-thawing cycles. Besides the presence of IL-33 we tested levels of IL-1α and IL-1β. In our experimental model, we used A549 cell line (alveolar type II-like cells), THP-1 promonocytic...
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Immunologic profile of experimental autoimmune encephalomyelitis
Novosádová, Iva ; Fišerová, Anna (advisor) ; Zajícová, Alena (referee)
5 Anglický abstrakt Experimental autoimmune encephalomyelitis (EAE) is widely accepted as a murine model of human multiple sclerosis autoimmune disease. Murine EAE is usually actively induced by immunization with a suitable myelin antigen. Following immunization, CD4+ T helper lymphocytes Th1 and Th17 accumulate in the nervous tissue and via the production of cytokines, they mediate an inflammatory reaction and the subsequent destruction of myelin. The main goal of this study was the induction of EAE with clinically observable symptoms and the observation of changes in the counts and phenotypes of cells, mainly NK and T cells. NK cells express a wide range of inhibitory and activation receptors from the C-lectin-like receptor superfamily. The specific ligand of the activating NKR-P1C isoform is still unknown and thus this receptor's involvement in EAE was also observed. Another goal was the use of medication with regard to the disease progress improvement. For the purposes of this study, two inbred murine strains with distinct NKR-P1 surface expression were used - the SJL/J strain (expressing inhibitory NKR-P1B) and C57BL/6 (expression activating NKR-P1C). SJL mice elicited a relapse-remitting of EAE, while C57BL/6 had chronic EAE. Both mouse strains exerted changes in the counts of NK...
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Evolutionary implications of innate immunity receptors polymorphism
Bainová, Zuzana ; Vinkler, Michal (advisor) ; Zajícová, Alena (referee)
Interactions between hosts and their parasites are considered to be one of the major forces driving animal evolution. It can be assumed that the evolutionary changes will occur especially in host molecules directly involved in these interactions. The first line of host defense is formed by innate immunity receptors among which also pattern recognition receptors (PRRs) belong. PRRs detect the presence of parasites at the beginning of their invasion by binding characteristic structures of their bodies (so called pathogen-associated molecular patterns, PAMPs, e. g. lipopolysaccharide, flagellin or peptidoglycans) or abnormal self molecules (damage-associated molecular patterns, DAMPs, e.g heat shock proteins). Although this mechanism of immune system activation is based on the recognition of ligands that are relatively evolutionarily conservative in pathogens, growing body of evidence suggests that PRRs are highly polymorphic on both interspecific and intraspecific level. High frequencies of minority alleles can be observed in most populations studied. It has been proven that particular alleles of many PRRs may associate with increased or decreased resistance to various infectious or autoimmunity diseases. Relationship between polymorphic receptor and a disease could be the main force, which shapes the...
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