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The signifikance of extrapituitary prolactin and innate immune reaction in severe immunopathological conditions.
Chromá, Věra ; Daňková, Pavlína (advisor) ; Gabalec, Filip (referee)
Introduction: Communication between neuroendocrinne and immune system is arranged by hormones and cytokines in endocrinne, paracrinne and autocrinne manner. One of the factors involved is also prolactin, a pituitary hormone and an immune cytokine. Sepsis is a system reaction to inflammation mediated by Th1 immune response, which is supported by prolactin as well. Primary protection against sepsis is mediated by innate immunity. Toll- like receptors distinguish molecules, which are connected with pathogens. Afterwards this identification of a specific pathogen toll-like receptors trigger immune reaction with the main goal of destroying this pathogen and also with the goal of renewing the balance of the organism. It is supposed that in the organism that is hardly attacked by a pathogen, the PRL, TLR2 and TLR4 gene expression is on the increase. We studied the levels of PRL, TLR2 and TLR4 mRNA production in circulating monocytes derived from septic patients. Simultaneously, the effect of PRL -1149 G/T SNP on physiological levels of PRL mRNA and its expression in the course of sepsis was evaluated. Materials and methods: As a source of monocytes, blood specimens from 43 septic patients and 40 healthy controls were used. The blood of septic patients was taken three times with some time difference and...
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The use of thromboelastography (TEG) in the evaluation of coagulation in patients on intensive care unit (ICU)
Durila, Miroslav ; Cvachovec, Karel (advisor) ; Stříteský, Martin (referee) ; Kovářová Kudrnová, Zuzana (referee)
Patients in the intensive care unit are in critical condition which is often accompanied by a coagulation disorder. Sepsis as a leading cause of death in critically ill patients may be associated with both hypercoagulable state with microtrombi formation in microcirculation and with increased production of endogenous heparinoids with inhibitory effects on blood clotting. Central venous catheter and arterial catheter are established in patients for hemodynamic monitoring and these are flushed with heparin to prevent their closure. Both inputs are used for blood sampling for laboratory tests such as blood count and coagulation parameters, including thromboelastography (TEG). In the first step of the work, arterio-venous differences in coagulation parameters were investigated in patients with sepsis. Higher concentration of D-dimers and lower antithrombin activity were found in venous blood. This finding can be explained by increased antithrombin consumption in hypercoagulable state and reactive hyperfibrinolysis. Inconsistency in the site of blood sampling may then lead to misinterpretation of the pathophysiological processes in the body. No significant differences were found in TEG parameters. In the second step of the work we examined how heparin commonly used for catheter flushing affects TEG-assessed...
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Significance of prolactin as peripheral cytokine in dysbalance of immune system
Janatová, Kateřina ; Černá, Marie (advisor) ; Holáň, Vladimír (referee)
Background: Interactions between the neuroendocrine and immune system play an importatnt role in maintaining homeostasis. This communication is mediated by cytokines, neurotransmiters and hormones through endocrine, paracrine and autocrine signaling. Prolactin (PRL), hormone of anterior pituitary, is produced by a number of other tissues and cells of immune system. On periphery, PRL is cytokine. Sepsis is an inflamatory response of the organism to severe infection, Th1 immune response is activated and PRL could participate in it. Toll-like receptors (TLR) play a key role in a recognition of bacteial components and mediate a systemic response (with PRL secretion) during infection. It is supposed that activated immune system leads to increasing of PRL, TLR2 and TLR4 gene expression. We detected PRL, TLR2 a TLR4 mRNA levels in monocytes from patiens with system inflammation. We studied influence of single nucleotide polymorphism (SNP -1149 G/T) in PRL gene promotor, it supposed that G allele increases PRL expression. Materials and Methods: For the pilot study 30 patients diagnose with severe infectious event. Collectoin of patiens blood samples was performed consequently three times. Control group comprised 40 healthy individuals. One blood sample was taken from each healthy subject. For testing of...
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The Immune response in critically ill patients
Melichová, Jana ; Průcha, Miroslav (advisor) ; Zazula, Roman (referee)
Cardiosurgery is a potent activator of the immune response similar to sepsis in critically ill patients. Therefore the differential diagnostics of infectious and non-infectious etiology plays an important role. The aim of the study was to compare the biomarkers in the diagnostics of patients after cardiosurgery and in septic patients. 24 patients fulfilling criteria of sepsis and 8 patients after cardio surgery were involved in the study. We demonstrated higher specificity and sensitivity of PCT in comparison with CRP at diagnostics of sepsis. sTREM-1, expression of TREM-1 on monocytes, TREM-1 on granulocytes and the number of FoxP3+T regulatory lymphocytes do not provide a reliable differential diagnostics of infectious and non-infectious etiology of examined group of the patients. The number of lymphocytes in patients with sepsis is a useful parameter at this diagnostics. Cardiosurgery represent a significant impulse for the evocation of the systemic inflammatory response of non-infectious etiology.
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Problem of sepsis in the intensive care unit
Škrabánková, Hana ; Marková, Eva (advisor) ; Jankovcová, Kateřina (referee)
The aim of the thesis was to determine the microbial colonization at the injection site of central venous catheter and its possible influence to maintain the lowest incidence of catheter sepsis. The culture results of skin swabs and taken blood cultures from two groups of patients were compared with each other using qualitative analysis methods. Those were Intensive Care Unit patients with established central venous catheter. The result was the discovery of microbial colonization of the skin and the representation of different pathogenic strains at the site of a central venous catheter for both established groups. Further showed that the higher the risk of microbial colonization with symptoms of catheter sepsis was seen where patients have central venous catheter established for longer period of time and have been treated under home parental nutrition program with stoma. The recommendation is the continuous education of health professionals to act in accordance with the latest preventive treatment recommendations, and the emphasis is mainly placed on hands cleaning and disinfecting as the hands of health professionals are one of the most common and risky way of transmission of pathogenic strains with potential nosocomial infection formation. Aseptic principles must be repeatedly communicated to all...
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Hemo elimination method in the treatment of sepsis and early multi-organ dysfunction
Sýkora, Roman ; Matějovič, Martin (advisor) ; Chytra, Ivan (referee) ; Maňák, Jan (referee) ; Šrámek, Vladimír (referee)
The most common cause of death in patients with sepsis/septic shock is deterioration of the function of multiple organs, termed multiple organ dysfunction syndrome. Although our understanding of mechanisms involved in the pathophysiology of sepsis-induced tissue damage has improved substantially, therapy of these syndromes still remains largely supportive. The hallmark of sepsis is an overwhelming systemic production of both pro- and anti-inflammatory mediators leading to generalized endothelial and epithelial damage, microcirculatory-mitochondrial distress, altered endocrine and coagulation homeostasis and cellular immune hyporesponsiveness. Therefore, the hypothesis that modulation of this excessive immunological and biological response to infection might improve patient outcome appears reasonable. Hemoelimination techniques represent biologically plausible way to provide non-specific removal of soluble pro- and anti-inflammatory mediators, although the concept of blood purification in sepsis remains a matter of considerable debate. The aim of this thesis was to elucidate effects of 1) high volume hemofiltration (HVHF) and 2) coupled plasma filtration adsorption (CPFA) in a long-term, hyperdynamic porcine septic shock model, which fulfils the criteria for human sepsis. We hypothesized that both HVHF and...
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Bacteriological examination of blood cultures
MAJEROVÁ, Natálie
The aim of this bachelor thesis is the introduction of possibilities of hemoculture examination, assuming the methodology of evolving the manual and automated hemocultures in tha Stafila laboratories and at the Department of Bacteriology in the Hospital of České Budějovice. Then it deals with the comparison of those two systems and learning the basic interpretation of results. In the theoretical part I focus on blood circulation infections, sepsis and its diagnostics and nozocomial infections.The last part delas with the most frequent bacterial causers of blood circulation infections. The practical part focuses on the approach of hemoculture process, firstly on the preanalytic part which includes the sample collection and its transport. Secondly there is the analytic part where cultivation bottles are described as well as the basic methods for bacteria certification such as making the microscopic sample and bacteria cultivation. The next part delas with the determination of a bacteria kind and its antibiotics sensitivity.In the last part there are results of cultivation of the followed up file from the both methods. Positive findig occuresd in 15% of taken hemocultures.
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Microbiological examination of blood cultures by automated system BACTEC
VANIŠOVÁ, Petra
Objective: The aim of this work is to describe a presence of pathogenic microorganisms in bloodstream that could possibly threaten the health and life of a patient. Using an automated microbial detection system Bactec we have detected bacteria or yeast that partake on an extension of a sepsis in given area, we have also compared results with special literature. We have evaluated the percentage of contaminated samples trying to point out the importance of a correct material sampling and also the importance of the level of education of the medical staff. After comparing samples in the cell culture medium we have used control skin smears. We have also marked the period of time for which the cultivation occurred since it is an important factor in the restoration of a patient and lowers the cost of a prospective treatnent. The method: In contrast with the theoretical section of this this work, the practical part is focused on laboratory samples preparation and actual analysis, including identification of the microorganisms, using microscopic, cultivation and biochemical techniques. For the detection of infective agents in blood culture we have used automated microbial detection system BACTEC 9050 which uses fluorescent sensor for the registration of CO2 creation. Carbon dioxide reacts with water and creates acid. This chemical process is followed with the descrease of pH. The content of a positive aerobic vial is vactinated for the right cultivation broth. Microbial groth is then evaluated after 17 to 24 hours in thermostat, followed be antibiotics sensitivity tests and then the optimal treatment is determined. The result: We have colected 1643 samples, 1359 of which did not show any pathogenic species. The work deals with findings inside of 284 samples which the machine marked as positive. The most common was Escherichia coli. The second place in quatity was represented by Klebsiella pneumoniae. The majority of bacteria were gram-negative (72 %). Sample contamination occured in 4.7 % of all samples and was connected with sampling errors. The conclusion: There is an annual growth in bloodstream infections that are also connected with a high percentage of death possibility. Blood cultivation examination with a use of automated systems is crucial for quick and precise detection of microorganisms.
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The problem of blood collecting for testing blood culture.
NOVÁKOVÁ, Olga
Current situation: Blood culture is intended to detect infection in the bloodstream. Increasingly today, complications of diseases like toxicemia, bacteremia and sepsis occur.Diagnosis of sepsis is not easy and blood culture plays an irreplaceable role. Blood sampling for blood culture is one of the technically challenging and time-consuming procedures, and these samples must be perfectly controlled by the nurses.Point of interest: Risk factors identification that may affect the results of blood culture relating to the collection of blood cultures was the main aim of the research. Determination of nurses' knowledge and compliance with the recommended procedures for collecting blood cultures, and identifi cation of the most frequently committed sisters' errors was the main objective.Participants: The research sample was chosen on purpose, with respect to the research problem. Group consisted of nurses working in the Department of Infectious Diseases in Hospital České Budějovice.Results: The recommended procedures of blood culture collection and risk factors that affect the result of blood culture are reviewed.The results of observations showed the errors of nurses taking blood culture are most often perpetrated.Nurses' knowledge is sufficient regarding the recommended sampling procedure and rules characterizing this collection, as detected by conducting interviews. Nurses are apprised with the procedure, and are able to justify different methods.Conclusion: Patients' lives could be saved by the correct diagnosis of bloodstream infections. The nurses have a high responsibility to perform their procedures in accordance with the sandards possible to eliminate risk factors that may affect the outcome of blood culture examination. The outcome of this work is to identify the most common errors made by the nurses and to propose preventive measures that would eliminate exposed areas of risk. The errors may be caused by ignorance, the automated process and also different time and mental pressure in patient care.
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