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Activity of antioxidant enzymes in different pathophysilogical states.
Vávrová, Lucie
Background: Oxidative stress is supposed to be implicated in the pathogenesis of several diseases which are connected with increased formation of reactive oxygen and nitrogen species (RONS). Oxidative stress could play an important role in the pathogenesis of inflammation and sepsis, acute and chronic pancreatitis or in the development of cancer. Organisms are protected against RONS from antioxidant system that is composed of antioxidant enzymes and non-enzymatic antioxidants. To the most important antioxidant enzymes belong superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase, glutathione reductase and paraoxonase (PON). The aim of this Doctoral Thesis was to investigate the behaviour of three of these antioxidant enzymes - CuZnSOD, CAT and PON1 in different pathophysiological states. Materials and methods: The activities of CuZnSOD, CAT and PON1 were measured in six different pathophysiological states. Forty patients with metabolic syndrome (MetS), 35 women with depressive disorder (DD), 30 septic patients (SP), 50 patients with pancreatic cancer (PC), 50 patients with chronic pancreatitis (CP) and 13 patients with acute pancreatitis (AP) were included in different studies together with sex- and age-matched healthy controls (CON). Patients with AP and SP were observed in the course...
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Glutathione and glutathione-dependent enzymes in various pathophysiological states.
Kodydková, Jana
Backround: Oxidative stress (OS) has been implicated in pathogenesis of human disorders such as depressive disorder, sepsis, cardiovascular disease, acute and chronic pancreatitis, and cancer. Increased OS is result of imbalance between increased reactive oxygen and nitrogen species (RONS) production and / or insufficient activity of antioxidant defence system. Antioxidant system, which is composed of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidases (GPX), glutathione reductase (GR) and non- enzymatic antioxidant reduced glutathione (GSH) plays an important role in the protection of cells against enhanced OS. The aim of this study was to assess the OS markers and antioxidant enzymes in different pathophysiological states. Materials and methods: Activities of erythrocyte glutathione peroxidase (GPX1), GR and concentration of GSH as well as levels of OS markers were analysed in six different pathophysiologic states. These parameters were measured in 35 women with depressive disorder (DD), 40 patients with metabolic syndrome (MetS), 30 septic patients (S) followed up in the course of sepsis; 15 non-septic critically ill patients (NC), 13 patients with acute pancreatitis (AP), 50 with chronic pancreatitis (CP) and 50 patients with pancreatic cancer (PC), compared to...
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Replacement and Support of Vital Organs in Sepsis Pathophysiology and New Aspects of Treatments
Martínková, Vendula ; Matějovič, Martin (advisor) ; Chytra, Ivan (referee) ; Charvát, Jiří (referee)
Infectious diseases are the worldwide leading cause of morbidity and mortality. Sepsis is the major cause of death in infectious diseases. It is one of the most serious and also one of the most difficult treatable conditions of contemporary medicine. Sepsis is the main cause of death in intensive care units. Causal therapy of sepsis does not yet exist. With a far better understanding of patho/physiological mechanism of sepsis, it is possible to model new preclinical experiments to verify the efficiency and security of new therapeutic procedures. Large animal experiments in progressive sepsis, with the use of domestic porcine, play a vital role. Long-standing experience with this model and similarity to human facilitate the realization of more complex experiments with potential for the relevant translation of results into the subsequent clinical studies on human subjects. The objective of this doctoral dissertation was to assess on the clinically relevant model: 1) the efficiency and security of extracorporeal membrane oxygenation in the event of vasoplegic septic shock; and 2) the benefit of two innovative therapeutic approaches to treatment of sepsis: a) the intravenous administration of mesenchymal stem cells; and b) the activation of the neuro-inflammatory reflex through the vagus nerve stimulation.
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Endothelial Glycocalyx - Diagnostic Approach and Intervention Assesment
Pouska, Jiří ; Beneš, Jan (advisor) ; Málek, Jiří (referee) ; Štourač, Petr (referee)
UNIVERZITA KARLOVA Lékařská fakulta v Plzni Dizertační práce Endothelial glycocalyx - diagnostic approach and intervention assessment MUDr.Jiří Pouska ABSTRACT Endothelial glycocalyx (EG) is fine structure on the surface of endothelium. After extensive research in past years, revisited Starling principle was finally formulated. It describes fluid physiology in capillaries precisely. EG has pivotal role in keeping endothelium semipermeable and thus avoiding extensive filtration of fluids to interstitium. Assessment of EG is clinically difficult. Many pathological conditions lead to damage of EG (sepsis etc.). Intravenous fluid therapy is mainstay of treatment of such conditions. Our aim was to determine the changes of EG integrity depending on the choice of intravenous fluid and its infusion time in physiological and pathological conditions. Key words: Endothelial glycocalyx, infusion therapy, anaesthesia, sepsis, microcirculation.
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Sepsis and Sepsis-associated Acute Kidney Injury: Molecular Mechanism and Lovel Aspects of Treatment
Horák, Jan ; Matějovič, Martin (advisor) ; Šrámek, Vladimír (referee) ; Balík, Martin (referee)
Sepsis is the most common cause of death in the intensive care units worldwide. Despite the undeniable progress in pre- and clinical research, the effective causal therapeutic strategy still does not exist. Given to extremely complex and heterogenic host response to presence of infection, the paradigm "one disease, one drug" is obviously flawed and combination of multiple targets that involves early immunomodulation and cellular protection are needed. Cellular therapy using mesenchymal stem cells represents strategy that brought positive results in experiments with rodent septic models. Part of this thesis is dedicated to evaluation of safety and efficacy of stem cells intravenous administration to well-established large animal model of progressive peritoneal sepsis. Affecting of the neuroinflammatory reflex through the vagus nerve stimulation showed potential to attenuate sepsis in rodents. Rest of the text is dedicated to evaluation of vagus nerve stimulation effectivity in our model. KEYWORDS Sepsis, sepsis-associated acute kidney injury, mesenchymal stem cells, vagus nerve stimulation
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Analysis of volatile organic compounds produced by monocytes during sepsis
Bártová, Adéla ; Řezáčová, Veronika (referee) ; Mravcová, Ludmila (advisor)
This thesis is focused on the possibility of analysis of volatile organic compounds produced by monocytes during sepsis. Method of comprehensive two-dimensional gas chromatography with mass spectrometric detection was chosen for this purpose. Content of the first part was the optimization of the method of two-dimensional gas chromatography for the determination of volatile organic compounds. In this part were gradually adjusted parameters of the gas chromatography method to achieve the maximum efficiency. Further were adjusted conditions of samples preparation. Content of the second part was the usage of already optimized method for the analysis of the samples set of monocytes. Samples were subjected to the action of different inhibitors of the immune system and stimulators simulating bacterial or yeast infection. Based on this analysis were identified some compounds, which are produced by monocytes under condition simulating the infection.
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The basic microbiological examination of blood cultures in the automated blood culture system
ŽÁKOVÁ, Hana
This bachelor thesis is focused on examination of blood cultures in automatic blood culture system. In the theoretical part, it deals mainly with general terms of the presence of bacteria in the blood and the development of sepsis, as well as with the description of the most frequent infectious agents in blood cultures in different groups of patients and also with different types from the oldest to newest blood culture systems. Manual systems are described, such as Septi-Chek and Oxoid Signal System, and automated BACTEC and Bact / Alert systems that work on different principles. Also discussed is the subsequent processing of samples to identify microbial agents and determine their susceptibility to antibiotics. The aim was to acquire practical processing of blood cultures in automatic BACTEC hemocultivation system. The methodical part is devoted to the preparation and processing of samples for hemoculture, the subsequent procedure of inoculation on the cultivation soil and the preparation of a microscopic preparation stained by Gram. Furthermore, the thesis deals with the statistical processing of data provided at the microbiological department of the Nemocnice Písek, a.s. The statistical survey was carried out in order to determine the microbial spectrum in blood cultures and the percentage of positivity to the total number of collected blood cultures in 2014-2018. A total of 9136 blood cultures from 3027 patients were examined during the study period. Of the total number, 1663 were positive, i.e. 18 %. 82 % of blood cultures were negative, did not show growth of microorganisms. The microbial spectrum is very wide, with Escherichia coli, Staphylococcus aureus and Klebsiella species.
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Diagnostics of the blood cultures from the departments of internal medicine in Nemocnice Tábor, a.s.
SMRŽOVÁ, Kamila
The thesis describes blood culture testing at internal medicine departments of Nemocnice Tábor, a.s. (Tábor Hospital). The theoretical part presents the most important terms related to infections of the bloodstream and sepsis. EARS-Net, an important European project is also mentioned. The practical part describes blood culture laboratory process in all phases (preanalytical, analytical and post-analytical) including the steps taken when a sample is positive. Furthermore, the study evaluates data on blood cultures done at internal medicine departments of Nemocnice Tábor, a.s. in 2018, determining the incidence of gram-negative bacteria in positive samples and seeking to describe the differences between community and hospitalised patient groups. Finally, the study also gives an overview of antibiotic resistance in the most commonly detected gram-negative rods including a comparison to EARS-Net data (mean value for internal medicine departments in the Czech Republic). In total, 527 blood culture vials were tested as positive in 2018. A total of 267 positive vials were analysed further after the removal of duplicities and contaminations; a total of 97 strains of gram-negative rods were cultured from the vials. The most commonly detected gram-negative bacterial strains found in positive samples included Escherichia coli (48.5%) and Klebsiella pneumoniae (22.7%). Differences between community and hospitalised patient groups were found only for Klebsiella pneumoniae; however, the pathogens spectra in both groups were not distinctively different. In terms of resistance assessment in the two most common bacterial strains, a higher frequency of incidence of Klebsiella pneumoniae strains resistant to third generation cephalosporins and ciprofloxacin is alarming, also in comparison to EARS-Net data (mean value for internal medicine departments in the Czech Republic).
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The basic microbiological examination of blood cultures in the automated blood culture system
ŽÁKOVÁ, Hana
This bachelor thesis is focused on examination of blood cultures in automatic blood culture system. In the theoretical part, it deals mainly with general terms of the presence of bacteria in the blood and the development of sepsis, as well with the description of the most frequent infectious agents in blood cultures in different groups of patients and also with different types from the oldest to newest blood culture systems. Manual systems are described, such as Septi-Chek and Oxoid Signal System, and automated BACTEC and Bact / Alert systems that work on different principles. The subsequent processing of samples is also discussed to identify microbial agents and determine their susceptibility to antibiotics. The aim was to acquire practical processing of blood cultures in automatic BACTEC hemocultivation system. The methodical part is devoted to the preparation and processing of samples for hemoculture, the subsequent procedure of inoculation on the cultivation soil and the preparation of a microscopic preparation stained by Gram. Furthermore, the thesis deals with the statistical processing of data provided at the microbiological department of the Nemocnice Písek, a.s. The statistical survey was carried out in order to determine the microbial spectrum in blood cultures and the percentage of positivity to the total number of collected blood cultures in 2014-2018. A total of 9838 blood cultures were examined during the study period. Of the total number, 2338 were positive. That is 24%. 76% of blood cultures were negative, did not show growth of microorganisms. The microbial spectrum is very wide, the most common infection agents are Staphylococcus plasmakoagulasa negative, Escherichia coli, Staphylococcus aureus subsp. aureus, Staphylococcus epidermidis and Klebsiella species.
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