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Lipid emulsions in lonf-term parenteral nutition
Ševela, Stanislav ; Novák, František (advisor) ; Cahová, Monika (referee) ; Lahoda Brodská, Helena (referee)
The combination of intestinal failure and parenteral nutrition is a condition associated with mild chronic inflammation and organ injury, the most significant of which is hepatic injury. This crossover-controlled trial investigated the effects of different intravenous lipid emulsions (IVLE) enriched with increased doses of omega-3 polyunsaturated fatty acids (n-3 PUFA) from fish oil (FO) in patients on home parenteral nutrition (HPNP) for chronic intestinal failure. Twelve patients were consecutively given three different IVLEs, with an FO emulsion added after each IVLE. The study also included twelve paired healthy controls for comparison. The aim of this study was to investigate how long-term administration of these IVLEs affected primarily the erythrocyte and plasma fatty acid (FA) spectrum, as well as the inflammatory response, antioxidant status, markers of liver function and bile acid spectrum in HPNP, and how these changes differed from healthy subjects. We demonstrated successful incorporation of n-3 PUFA into erythrocyte phospholipids and corresponding changes in the plasma FA spectrum, characterized in particular by significant increases in n-3 versus n-6 PUFA, with a decrease in the n-6/n-3 PUFA ratio. We also found a more prolonged effect on these changes with high dose FO...
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Influence of the citrulination of histon proteins on the expression of selected genes in myeloid cells
Tučková, Kristýna ; Dobeš,, Pavel (referee) ; Číž,, Milan (advisor)
Neutrophils are major cell type of innate immunity, that can eliminate pathogens by different mechanisms. One of these mechanisms is called NETosis, which leads to release of decondensed chromatin and citrullinated histone proteins. Citrullination is post-translational modification catalysed by peptidylarginine deiminase (PAD) and causing transformation of possitively charged arginin to neutral citrullin and can change expression of cytokine genes. Concetrations of pro-inflammatory cytokines (IL-8, TNF, IL-1) were measured after activation of PAD4 and induction of citrullination. Calcium ionophore was used to induce citrullinaton, Cl-amidine and TDFA were used as inhibitors. Production of cytokines was assessed by ELISA on protein level and by qPCR on mRNA level. It was found that induction of citrullination led to increased concentrations of IL-8 and IL-1. Elevated gene expression of IL-8 was confirmed on mRNA level. Both inhibitors were able to decrease level of histone H3 citrullination and IL-8 and IL-1 concentrations. Expression of TNF was not detected on protein and mRNA level.
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Monitoring of sublingual allergen immunotherapy using serum cytokine levels
Brzáková, Adéla ; Řezáčová, Vladimíra (advisor) ; Andrýs, Ctirad (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Bc. Adéla Brzáková Supervisor: Mgr. Vladimíra Řezáčová, Ph.D. Title of thesis: Monitoring of sublingual allergen immunotherapy using serum cytokine levels Keywords: allergen immunotherapy, cytokine, ELISA, grass pollen allergy Background: The aim of this thesis was to find out whether there are detectable changes in the levels of IFNγ, IL-4 and IL-5 during allergen immunotherapy and whether these cytokines could thus serve as biomarkers of the efficiency of this treatment. Methods: Selected cytokines were determined using Quantikine® HS ELISA Human IFN-γ Immunoassay, Quantikine® HS ELISA Human IL-4 Immunoassay and Quantikine® ELISA Human IL-5 Immunoassay. The monitored population consisted of 32 polinotics, 19 of which were administered allergen immunotherapy and 13 of which constituted the control group. As part of the follow-up, both groups also continuously recorded the severity of their symptoms using a visual analog scale. Results: In the group of treated patients, a decrease in IL-4, IL-5 and IFNγ concentrations was observed during treatment, but no significant differences in the values of these cytokines were found between patients treated with Oralair and control individuals. Using...
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Produkce cytokinů v průběhu mastitid skotu
Tylčová, Natálie
The aim of this work was to detect the pro-inflammatory cytokine TNF-α and the anti-inflammatory IL-10. The experiment included 20 dairy cows at different stages of mastitis, which came from the Nová Ves agricultural cooperative. 80 samples were totaly taken, from all quarters, from each dairy cow, which were divided into three groups. The first group represented healthy dairy cows, without clinical signs of mastitis, with a somatic cell count of up to 100 000, from 5 dairy cows, that is 20 milk samples. The second group consisted of dairy cows with clinical signs of mastitis and somatic cell counts up to 400 000, from 10 cows, that is 40 milk samples. The third group consisted of dairy cows with somatic cell counts of more than 1 000 000, from 5 cows, that is 20 milk samples. These samples were processed in the laboratory of the Department of Animal Morphology, Physiology and Genetics at Mendel University in Brno. The concentration of TNF-α and IL-10 was detected by ELISA. The average concentration of TNF-α in the first group was 138.70 pg/ml, in the second group 133.60 pg/ml, in the third group 95.05 pg/ml. The average concentration of IL-10 in the first group was 9.52 pg/ml, in the second group 11.03 pg/ml, in the third group 8.66 pg/ml.
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Pathophysiological and genetic factors affecting serum uric acid level.
Hasíková, Lenka ; Závada, Jakub (advisor) ; Hrnčíř, Zbyněk (referee) ; Horák, Pavel (referee)
Introduction: Serum uric acid level (SUA) depends on the balance between its production and excretion. SUA is associated with several transmembrane proteins responsible for reabsorption (mainly URAT1 and GLUT9) and secretion (ABCG2) on the apical and basolateral membranes of the proximal tubules in the kidney, and in the case of ABCG2, it also correlates with its significant excretion through the gastrointestinal tract. Gout is a metabolic disease caused by the deposition of urate crystals in the joints and tissues. Chronic hyperuricemia is a primary risk factor for the development of gout; however, gout patients usually have a lower SUA during an acute gout attack than in the intercritical periods. The exact mechanism of this phenomenon is unknown. It has been speculated that the systemic inflammatory response can explain this discrepancy. The aim of the study is to determine whether treatment with specific inhibitors of the proinflammatory cytokine TNF (TNFi) affects SUA in patients with systemic rheumatic disease (SRD), and whether changes in SUA correlate with changes in selected proinflammatory cytokines or with the biomarker of oxidative stress, allantoin. Another aim is to determine the frequency and effect of allelic variants in the ABCG2 urate transporter gene in patients with primary...
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Circadian system as a modulator of neuroinflammation
Kotková, Eliška ; Spišská, Veronika (advisor) ; Dočkal, Tereza (referee)
The circadian system is involved in the regulation of biological rhythms in physiological, behavioural and immune processes. These rhythms can be found in the central nervous system, including the blood-brain barrier, astrocytes, microglia, and the pineal gland, which produces the hormone melatonin. Neuroinflammation is a complex response of the central nervous system to inflammatory stimuli by rhythmic expression of pro-inflammatory and anti-inflammatory mediators or by rhythmic regulation of immune system cells. Studies have examined the influence of genes and proteins of the circadian system, suprachiasmatic nuclei, melatonin, and glial cell rhythms on neuroinflammation. Lipopolysaccharide was used to induce neuroinflammation in these studies. Based on these studies, the effect of melatonin on mikroglia and endothelial cells, and the responses of suprachiasmatic nuclei was evaluted as the most important circadian modulator of neuroinflammation. This thesis describes the basic principles of the circadian system and neuroinflammation, with the last section presenting the modulation of neuroinflammation by the circadian system. Keywords: astrocytes, blood-brain barrier, circadian system, cytokines, immune system, melatonin, microglia, neuroinflammation, suprachiasmatic nuclei
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Cytokine networks and their impact on the immune profile of acute myeloid leukemia (AML) patients
Ptáček, Antonín ; Musil, Jan (advisor) ; Fišer, Karel (referee)
Acute myeloid leukemia (AML) is a malignant hematological disorder characterized by aberrant expansion of blasts in the bone marrow and peripheral blood. The immune system protects the body from leukemia by eliminating transformed cells. However, in AML, the abilities of immune cells are affected both by direct contact between leukemic cells and effector cells, as well as by cytokines, metabolites and other soluble proteins that, together with the cells, form the specific AML microenvironment. The effects of cytokines and other soluble molecules in the AML microenvironment are not sufficiently described yet. This thesis aimed to optimize and implement a multiparametric flow cytometry panel for the measurement of cell populations and to implement multiparametric assays for the analysis of cytokine levels, chemokines and other soluble proteins in plasma. The following goal was to use these methods to characterize the frequency and functional phenotype of cell populations and the levels of the soluble proteins and to describe their influence on disease severity and overall survival of the patients. We also tried to find novel biomarkers of the immune escape of leukemic cells. In patients, we observed a suppressive microenvironment with aberrant levels of soluble receptors and other proteins. This...
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Vnímavost plemen kapra obecného k onemocnění způsobenému kapřím edémovým virem
BALOCH, Ali Asghar
Fish diseases refer to any condition that affects the health and well-being of fish, whether in the wild or in captivity. Fish are vulnerable to a variety of diseases caused by bacteria, viruses, fungi, parasites, and other pathogens. The prevalence and severity of these diseases vary widely, depending on the species of fish, the environmental conditions, and the management practices. Fish have a complex immune system that allows them to respond to a variety of diseases. Their immune system includes both innate and adaptive responses, which work together to protect them from infections. Innate immunity is the first line of defense and includes physical barriers such as the skin, scales, and mucus, as well as cellular and humoral responses that can quickly recognize and respond to pathogens. These responses include the activation of phagocytic cells such as macrophages and neutrophils, the release of antimicrobial peptides, and the production of reactive oxygen species. Adaptive immunity, on the other hand, is a more specific and targeted response that is mediated by T and B lymphocytes. These cells can recognize and respond to specific pathogens, and can develop immunological memory, allowing for a faster and stronger response to subsequent infections. When a fish encounters a pathogen, its immune system is activated, and a series of responses are initiated. The exact response depends on the pathogen and the fish species, but generally involves the recruitment of immune cells to the site of infection, the activation of phagocytic cells to engulf and destroy the pathogen, and the production of cytokines to stimulate the immune response. Cytokines specially help to coordinate the immune response and ensure that the appropriate mechanisms are activated to fight the infection. The initial investigation of the thesis provides a comprehensive review of cytokine responses in common carp exposed to CEV, CyHV-3, SVCV and Aeromonas hydrophila infections. It aims to synthesize existing literature on immune responses, particularly cytokine expression, and highlights the essential role of cytokines as inflammatory mediators and their mechanisms in response to both pathogenic and non-pathogenic immunostimulants in common carp. The production of these cytokines may result in various symptoms and signs of infections, which can lead to the recruitment of inflammatory cells that play an important role in clearing up different infections. Furthermore, this study provides insights into the ways by which cytokines can be used to diagnose and combat disease in common carp. Further, the examination of the immune response of carp to CEV infection using gene expression profiling and qPCR was carried out. The expression patterns of eleven immune-related genes were monitored in four different carp strains. The expression patterns of several immune-related genes were found to be similar across all carp strains and genogroups, while some significant differences were observed in koi carp infected with both CEV genogroups. Additionally, a resemblance was observed in the expression pattern of several genes between KSD-resistant AS strain and koi. Furthermore, the observed increase in mRNA expression of several immune related genes indicates a broader mucosal immune response triggered by CEV. Overall, these results provide essential insights into the immunological response of carp to infectious diseases and may help to develop effective management strategies for preventing and controlling CEV disease in aquaculture.
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Mechanisms of immune response against kidney allograft
Švachová, Veronika
Kidney transplantation improves the prognosis and quality of life of patients with end- stage renal disease. However, the occurrence of rejection still represents a major complication that can lead to failure of allograft function. In our work, we have studied the phenotypic changes of subpopulations of B lymphocytes and monocytes, which represent one of the key players involved in immune responses against kidney transplantation. In the part of the research devoted to B lymphocytes, we have observed a different redistribution of individual B cell compartments in the peripheral blood during the first year after transplantation without significant differences between used immunosuppression. We have demonstrated that sensitized and rejected patients had a lower level of IgMhigh CD38high CD24high transient B cells. These results support the hypothesis of a protective role of transient B lymphocytes. Activation of monocyte subpopulations in peripheral blood in the posttransplantation perion was also subject of research. We have compared a cohort of patients with normal graft function with a group of patients with acute rejection. The level of "transient" monocytes and HLA-DR expression in a subset of "non-classical" monocytes did not decrease in patients with rejection, compared to the group with a...
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The Role of Cytokines in Support of Malignant Properties of Glioblastomas
Fortelková, Nikol ; Hodný, Zdeněk (advisor) ; Nováková, Zora (referee)
The tumor microenvironment of Glioblastomas (GB) is very complex and transforms throughout radiotherapy and chemotherapy, posing a favourable environment for the malignant properties of GB including invasivity, which promotes aggressive recurrency of the tumor. These changes in the tumor microenvironment are partly induced by cellular senescence and cellular response to stress, accompanied by a specific secretome, so called senescence-associated secretory phenotype (SASP). The SASP is distinct in its secretion of growth factors, components of the extracellular matrix and remodelling enzymes, but above all proinflammatory cytokines, which may contribute to the malignant properties of GB. This thesis reviews the current state of knowledge of the potential role of selected cytokines of the SASP (IL-6, IL-1β, IL-8, IL-1α, TGF-β, CCL2, TNF-α) in support of the malignant properties of GB. Key words: glioblastoma (GB), senescence, SASP, malignant properties, cytokines
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