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Analysis of selected physiological parameters of athletes during exercise
Ambrožová, Monika ; Kolářová, Jana (referee) ; Ronzhina, Marina (advisor)
Regular sport activity is considered as neccessary part of our lifes. Sport is reccomended for preventing various diseases, enhancing immunity system or for controlling weight and for enhancing conditions. However, how severaly is sport activity benefit for human health and when it becomes harmful for it? This work deals with possible negative effects of strenous excercise on human organism. In our case it is ultra-maraton and mountbiking. In following parts of this work it is tried to explaine metabolic changes and influance of strenous excercise on immunological and biochemical parametres. The parameters immunoglobulin A, M, leukocytes, creatinine, sodium and potassium ions and enzymes such as lactatedehydrogenase, creatinekinase and alanineaminotransferase were studied. The values of these parameters were obtained from the blood samples taken before and after the race. A 23 samples from a 24-hour cycling race and 24 run-of-the-run race samples per 100 km were examined. A matlab tool was created for statistical analysis of data. A huge increase in the number of leukocytes was found in both types of races. In the MTB, it is an increase from 5.75 ± 1.29 x 109 / l to 11.80 ± 3.08 x 109 / l, in the race from 4.99 ± 1.84 x 109 / l to 13, 20 ± 5.36 x 109 / l. IgA did not significantly decrease, on the contrary slightly increased. IgM values have not changed significantly in runners. Cyclists experienced a statistically significant reduction in the original amount of 60.23 ± 6.54 mg / dl to 52.60 ± 10.43 mg / dl. There were statistically significant changes in CK and LDH levels in the race. From the original CK values of 4.25 ± 2.74 kat / l, the activity of the enzyme increased by 50.09 ± 60.08 kat / l due to the race. LDH is not significantly different in the cycling race as well as in the ALT enzyme. In this case, there was no statistically significant change to either type of race. Concentration of potassium and sodium ions decreased significantly. In potassium, its concentration dropped from 5.61 ± 0.59 mmol / l to 4.57 ± 0.39 mmol / l. Sodium ions dropped from 138.22 ± 1.17 mmol / l to 137.00 ± 1, 75 mmol / l.
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Vývoj imunologických testů pro detekci nebezpečných bakteriálních patogenů
Šmídová, Lada
The aim of the thesis was to develop an immunoassay for the detection of dangerous bacterial pathogen, methicillin-resistant Staphylococcus aureus (MRSA). MRSA infections cause global problems in health facilities which provide acute and follow-up care in inpatient and outpatient parts. That is why early and rapid diagnosis and targeted thereapies are very important for the subjects. E. coli was purified using the QIAquick PCR kit Purificatin isolated bacterial constructs, which were subsequently purified and dialyzed. The recombinant protein PBP2a in different concentrations was applied to a nitrocellulose membrane in the form of lines. Furthermore was performed optimized blot-line method for the detection of specific antibodies against the recombinant antigen PBP2a in the classes IgG, IgA and IgM. Several different concentrations of the conjugate Goat Anti-human IgG-AP, Goat Anti-human IgA-AP or Goat anti-Human IgM-AP were used for the detection. The color intensity of each line of the strip was evaluated with Immunoblot software. The measured values were used to determine diagnostic sensitivity, specificity and overall diagnostic match. Further testing of precision was carried out under repeatability conditions (intra-assay) and reproducibility (inter-assay). Precision of the method was expressed by coefficient of variation. As the most suitable for the manufacture of a IgG kit was determined the concentration of the antigen PBP2a from 0.30 mg/ml to 0.45 mg/ml and the concentration of IgG conjugate from 1:1500 to 1:1800. For class IgA as the most appropriate antigen was determined concentration PBP2a from 0.40 mg/ml to 0.52 mg/ml and conjugate concentrations of IgA from 1:500 to 1:1000. The coefficient of variation under repeatability conditions for the entire range of the IgG class is 10.09 %, and for IgA is 8.91 %. Variation coefficient reproducibility conditions for the entire range of the IgG class is 9.23 % and for IgA is 9.60 %. Precision of the method under conditions of repeatability and reproducibility for classes IgG and IgA meets the criteria for the manufacture of a diagnostic kit. Titration results showed that particular batch of cards made of nitrocellulose membrane coated with antigen PBP2a must always be verified on the panel of reference samples and the values of concentrations (for both antigen and conjugate) should be set according to the needs, but differently for the class of immunoglobulins IgG and IgA.
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Experimental infection of Oryctolagus cuniculus with fluke Fascioloides magna
Melounová, Klára ; Kašný, Martin (advisor) ; Novobilský, Adam (referee)
Fasioloides magna is a trematode parasitizing in the liver parenchyma of ruminants. Its life cycle is associated with the humid environment and includes intermediate freshwater snail hosts from family Lymnaeidae. According to the ability of host to form a certain type of a pseudocyst during fascioloidosis, they can be,divided in three groups, specific definitive hosts (red deers, fallow deers, roe deers), nonspecific definitive hosts (cattle, wild boars and elks) and atypical hosts (sheeps and goats). Beside the natural infections also the experimental infections of other potential host species has been realized (chamois, llama and bighorn sheep and traditional laboratory animals such as mice, guinea pigs, rats and rabbits). In the context of different diseases, many changes in infected organism can occur. These can be qualitatively and quantitatively evaluated. Similarly, during fascioloidosis the changes associated with the presence of the parasite in the host's body is possible to monitor, e.g. antibody production, increase in the number of eosinophils, release of eggs in faeces, internal bleeding, or the level liver damage. The liver damage is corresponding primarily to biochemical parameters of blood, not only the liver enzymes, but also other blood components, like blood proteins, lipids,...
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Analysis of selected physiological parameters of athletes during exercise
Ambrožová, Monika ; Kolářová, Jana (referee) ; Ronzhina, Marina (advisor)
Regular sport activity is considered as neccessary part of our lifes. Sport is reccomended for preventing various diseases, enhancing immunity system or for controlling weight and for enhancing conditions. However, how severaly is sport activity benefit for human health and when it becomes harmful for it? This work deals with possible negative effects of strenous excercise on human organism. In our case it is ultra-maraton and mountbiking. In following parts of this work it is tried to explaine metabolic changes and influance of strenous excercise on immunological and biochemical parametres. The parameters immunoglobulin A, M, leukocytes, creatinine, sodium and potassium ions and enzymes such as lactatedehydrogenase, creatinekinase and alanineaminotransferase were studied. The values of these parameters were obtained from the blood samples taken before and after the race. A 23 samples from a 24-hour cycling race and 24 run-of-the-run race samples per 100 km were examined. A matlab tool was created for statistical analysis of data. A huge increase in the number of leukocytes was found in both types of races. In the MTB, it is an increase from 5.75 ± 1.29 x 109 / l to 11.80 ± 3.08 x 109 / l, in the race from 4.99 ± 1.84 x 109 / l to 13, 20 ± 5.36 x 109 / l. IgA did not significantly decrease, on the contrary slightly increased. IgM values have not changed significantly in runners. Cyclists experienced a statistically significant reduction in the original amount of 60.23 ± 6.54 mg / dl to 52.60 ± 10.43 mg / dl. There were statistically significant changes in CK and LDH levels in the race. From the original CK values of 4.25 ± 2.74 kat / l, the activity of the enzyme increased by 50.09 ± 60.08 kat / l due to the race. LDH is not significantly different in the cycling race as well as in the ALT enzyme. In this case, there was no statistically significant change to either type of race. Concentration of potassium and sodium ions decreased significantly. In potassium, its concentration dropped from 5.61 ± 0.59 mmol / l to 4.57 ± 0.39 mmol / l. Sodium ions dropped from 138.22 ± 1.17 mmol / l to 137.00 ± 1, 75 mmol / l.
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Humoral response of specific hosts to bird schistosome antigens
Turjanicová, Libuše ; Mikeš, Libor (advisor) ; Kolářová, Libuše (referee)
This thesis focuses on humoral immune response of specific hosts to antigens of various developmental stages of bird schistosomes T. regenti and T. szidati, and follows up on previous research of antibody response in non-specific hosts (mouse, human). Sera of experimentally infected and hunted-down wild ducks were examined using the ELISA and western blot methods. The sera samples were taken in predefined intervals. Results of the ELISA analysis show the process of humoral immune response after infection by bird schistosomes. The level of specific antibodies IgY against homogenate of T. regenti cercariae increased significantly 20 d.p.i. in ducks infected by T. regenti. Such reaction wasn't observed in ducks infected by T. szidati. Slight changes in level of specific antibodies IgM against T. szidati cercariae homogenate were observed 10 d.p.i. only in fully immunocompetent ducks and in reinfected ducks. Examination of hunted-down wild ducks didn't prove infection by bird schisosomes; this conclusion was confirmed by results of the ELISA analysis. IgY antibodies from ducks infected by T. regenti demonstrated strong reactions with 2 antigens in ranges 49-47 kDa and 47-45 kDa. Other reactions, which were recognized, have not been observed in all specimen. An Western blott with homogenate from 7 days...
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