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Induction of innate immune response against intracellular bacterium Francisella tularensis
Sommerová, Gabriela ; Konečná, Klára (advisor) ; Myslivcová Fučíková, Alena (referee)
Background: The purpose of this bachelor thesis is to describe Francisella tularensis and activation of innate immunity during host infection. Main findings: The main findings of the work include the method of recognition of F. tularensis via Toll-like receptors, the production of cytokines and chemokines and the subsequent involvement of other components of innate immunity, including neutrophils, NK cells, or humoral components of the immune response, which also includes complement. The information is based on a number of studies performed mainly on mouse models. Conclusions: The intracelullar bacterium F. tularensis is known to cause tularemia. Its high infectivity, together with the high risk of death in the lung form, raises great concerns about the misuse of this bacterium as a biological weapon. To date, not all of its mechanisms of pathogenesis are known, which is a major problem in the development of effective vaccines. The induction of innate immunity appears to be very important in the host's defense against F. tularensis. However, despite several decades of research, the mechanisms of F. tularensis involved in host cell manipulation, including the regulation of the induction of immune response to F. tularensis infection, have still not been fully elucidated. Key words: Francisella...
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Investigation of protein-protein interaction important for pathogenesis of tularemia infection
Prokšová, Magdaléna ; Stulík, Jiří (advisor) ; Myslivcová Fučíková, Alena (referee) ; Bouchal, Pavel (referee)
Francisella tularensis, sometimes called as a "stealth pathogen", causes zoonotic disease tularemia. Uniqueness of this intracellular bacterium is due to its ability to infect, survive and replicate within host phagocytic cells and evade the host immune response. Because of its extreme infectivity, ability to cause disease via inhalation route, and absence of a vaccine licensed for human use, F. tularensis is classified as a potent biothreat agent. The escape from phagosome plays a key role in the virulence of the bacterium. Type VI secretion system and F. tularensis pathogenicity island proteins are involved in this process; however, the exact molecular virulence mechanisms of F. tularensis are not fully characterized yet. The aim of this work was to characterize host-pathogen protein-protein interactions, which direct the infection process to F. tularensis benefits. The minimal experimental approach was selected for identification of protein-protein interaction between the host and the pathogen. The selected secreted proteins from FPI of F. tularensis subspecies novicida were fused with an epitope anchor FLAG tag and expressed in HEK 293T cell line. Interaction partners were identified by affinity purification followed by nanoLC-MS/MS analysis. The data indicate that the bacterial protein IglJ...
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Use of proteomic analysis to identify potential biomarkers for cardiovascular disease
Fučíková, Alena ; Lenčo, Juraj (advisor) ; Hernychová, Lenka (referee) ; Pudil, Radek (referee)
Hypertrophic cardiomyopathy is one of the most common inherited diseases of the cardiovascular system. Although this disease is known for a long time, a suitable diagnostic procedure uncovering its early stages in patients with negative or unknown family history is still lacking. A development of the method for targeted proteomic analysis in combination with subsequent quantification of chosen hypothetical markers of hypertrophic cardiomyopathy was the main aim of this thesis. This method is able to detect very small amounts of chosen markers in the minimum amount of complex biological material. Moreover, in combination with properly used standards, targeted proteomic analysis enables quite precise quantification of many analytes in a relatively short time. Several previously described protein markers of hypertrophic cardiomyopathy were assayed and quantified using unique proteomic technique. Concurrently, a new potential protein marker - soluble fibronectin - was described. Protein concentrations were validated using enzymoimmunoanalytical method and obtained results were compared with targeted proteomic analysis findings. In the presented thesis, a new short method was developed for detection and quantification of potential markers of hypertrophic cardiomyopathy. The main objectives of this...
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Induction of innate immune response against intracellular bacterium Francisella tularensis
Sommerová, Gabriela ; Konečná, Klára (advisor) ; Myslivcová Fučíková, Alena (referee)
Background: The purpose of this bachelor thesis is to describe Francisella tularensis and activation of innate immunity during host infection. Main findings: The main findings of the work include the method of recognition of F. tularensis via Toll-like receptors, the production of cytokines and chemokines and the subsequent involvement of other components of innate immunity, including neutrophils, NK cells, or humoral components of the immune response, which also includes complement. The information is based on a number of studies performed mainly on mouse models. Conclusions: The intracelullar bacterium F. tularensis is known to cause tularemia. Its high infectivity, together with the high risk of death in the lung form, raises great concerns about the misuse of this bacterium as a biological weapon. To date, not all of its mechanisms of pathogenesis are known, which is a major problem in the development of effective vaccines. The induction of innate immunity appears to be very important in the host's defense against F. tularensis. However, despite several decades of research, the mechanisms of F. tularensis involved in host cell manipulation, including the regulation of the induction of immune response to F. tularensis infection, have still not been fully elucidated. Key words: Francisella...
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Use of proteomic analysis to identify potential biomarkers for cardiovascular disease
Fučíková, Alena ; Lenčo, Juraj (advisor) ; Hernychová, Lenka (referee) ; Pudil, Radek (referee)
Hypertrophic cardiomyopathy is one of the most common inherited diseases of the cardiovascular system. Although this disease is known for a long time, a suitable diagnostic procedure uncovering its early stages in patients with negative or unknown family history is still lacking. A development of the method for targeted proteomic analysis in combination with subsequent quantification of chosen hypothetical markers of hypertrophic cardiomyopathy was the main aim of this thesis. This method is able to detect very small amounts of chosen markers in the minimum amount of complex biological material. Moreover, in combination with properly used standards, targeted proteomic analysis enables quite precise quantification of many analytes in a relatively short time. Several previously described protein markers of hypertrophic cardiomyopathy were assayed and quantified using unique proteomic technique. Concurrently, a new potential protein marker - soluble fibronectin - was described. Protein concentrations were validated using enzymoimmunoanalytical method and obtained results were compared with targeted proteomic analysis findings. In the presented thesis, a new short method was developed for detection and quantification of potential markers of hypertrophic cardiomyopathy. The main objectives of this...
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Influence hematological and biochemical parameters by different dogs.
Pechová, Eva ; Fučíková, Alena (advisor) ; Václav, Václav (referee)
The topic of this diploma work was to evaluate the hematological and biochemical indicators of various breeds of dogs. The aim of this work was to evaluate the blood samples taken
from 10 dogs and 10 bitches of various breeds and find out the dependency of values
in relation to their age, nutrition and sex. The samples were processed and evaluated from May to October 2015. The following was examined from the hematological indicators: RBC, HGB, HCT, MCV, WBC and a blood smear was made, from which a differential count
of leucocytes was made. Furthermore, the following biochemical indicators were examined: TP, GLU, CHOL and ATP. The literature research first focused on the basic components
of dog feed. Moreover, the hematological and biochemical indicators of blood as well as the methodology of the whole experiment were described. The results were processed with the aid of the RStudio program and the following basic statistical characteristics were counted: median, minimum, maximum and standard deviation. The ANOVA statistical method was used, in which the dependency of values among sex, age category and the type of feed fed
to the animals was determined. Furthermore, the values among the breeds were compared. From the differences observed among the breeds, there was an increase in red blood cells
in German Shepherds and Dobermans. Dobermans, Labrador Retrievers and German shepherd crossbreeds had increased values of ALT. Significant findings showed that there was an increase in the number of red blood cells and hematocrits in subjects fed with granulated feed as opposed to subjects with a combined diet. The difference was also observed for MCV between the genders.
Differences between the genders were found for ALT values. More data would be needed
to confirm the relationship to age. ALT values are dependent on the type of feed, where the values are higher in subjects fed with a completely granulated diet. The values of total protein increased in dogs with age. This trend is likely to be observed in bitches. For subjects fed a combined diet (granulated feed + home diet) an increase in the value of total protein was observed with age. For subjects fed granulated feed, this increase was not as significant.
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The influence of different diet on the hematological and biochemical parameters in rats
Hudíková, Tereza ; Fučíková, Alena (advisor) ; Jan, Jan (referee)
This thesis aims to investigate the relationship between increased protein dosage intake and selected haematological and biochemical markers. An experiment was designed using the Wistar laboratory rats of the male gander and similar age (born within +/- 5 days). These specimens were then divides into 3 groups with 10 specimens each. The age of the specimens at the beginning of the experiment was around 62 days. The first group was fed the standard corny mixture ad libitum, while the second group received 6 grams of the standard corny mixture and 10 grams of beef meat per day (thus reflecting the same amount of nitrous compounds as for the first group) and the last (third) group was fed ad libitum with homogenised beef meat. Before the diet was imposed on the three groups, blood samples were collected from all specimen from the lateral tail vein; after the diet was completed, blood samples were collected by in vivo heart puncture. All blood samples were subjected to haematological (using EDTA blood) and biochemical (using blood plasma) analysis. Overall, the following markers were measured: amount of erythrocytes, haematocrit level, haemoglobin level and Mean Corpuscular Volume (MCV) for the haematological analysis and triacylglycerol level, cholesterol level, amount of urea, amount of magnesium and the amount of iron for the biochemical analysis. Dietary induced significant changes were observed for the following markers: number of erythrocytes, levels of haemoglobin and MCV from the haematological analysis and cholesterol levels, amount of magnesium and amount of iron from the biochemical analysis; therefore showing that altered diet can have affect haematological and biochemical blood markers.
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