| |
|
Characterization and function of the transferrin from the tick \kur{Ixodes ricinus}
KADLECOVÁ, Jana
Iron metabolism is an important physiological process in ticks which has a great potential for an efficient tick control. This thesis is focused on the characterization and function assessment of the transferrin-related protein in the hard tick Ixodes ricinus. The full sequence Ixodes ricinus transferrin was determined and revealed that this protein is related to the insect transferrins of the type 2 (melanotransferrins). RNA interference silencing of tick transferrin did not affect tick feeding and survival suggesting that function of this protein is not directly involved in the iron transport.
|
|
Regulation of the expression of MHC class I molecules and other immunoactive molecules on tumor cells
Moravcová, Simona ; Vopálenský, Václav (referee) ; Reiniš, Milan (advisor)
4 Abstract Regulation of the expression of MHC class I molecules and other immunoactive molecules on tumor cells The aim of this master thesis project was to characterize the effects of IFNγ, TNFα and of the epigenetic agents 5-azacytidine (5AC) and 5-aza-2'-deoxycytidine (5AZAD) on the expression of molecules important for antigen presentation and modulation of the immune responses against tumors (MHC class I molecules and other immunoactive molecules CD54, CD80, B7-H1, B7-H2 and CD1d) on tumor cells. The experimental model used for this purpose were the HPV16-associated murine tumor cell lines, either MHC class I positive or negative. The goal was to determine the changes in surface expression of these molecules after treatment by FACS studies and also the expression at the mRNA level using qPCR. Expression of these proteins was also compared in the experiments in vitro on tumor cell lines and ex vivo on tumor cells explanted from animals. The most interesting result is the observation that 5AZAD increases the expression of B7-H1 which inhibits T cell mediated immune response and that 5AZAD and IFNγ act synergistically in the induction of the expression of MHC class I, CD54 and B7-H1 molecules. Klíčová slova: imunoeditace, protinádorová imunita, HVP16, MHC glykoproteiny I. třídy, B7-H1, IFNγ, TNFα,...
|
| |
| |
|
The study of molecular and biochemical bases of cytochrome c oxidase deficiency
Veselá, Kateřina ; Zeman, Jiří (advisor) ; Farghali, Hassan (referee) ; Drahota, Zdeněk (referee)
This thesis has been worked out in The laboratory for study of mitochondrial disorders (Department of Pediatrics, 1st faculty of Medicine, Charles Univezity in Praha), which serves as the diagnostic center for patients from Czech and Slovak Republics. During the last years, more than 40 children with isolated COX deficiency were diagnosed in our lab. But molecular background except 12 patients with mutations in SURF1 gene was remaining unknown. Due to the lack of adequate treatment for these patients, the genetic counseling and the possibility of prenatal diagnostics have high importance for the families. The possible dual origin of the defect with different hereditary aspects makes the genetic counseling in the affected families complicated and prenatal diagnostics based only on biochemical analyses very problematical if even possible. This work had been arisen basically from the necessity to find the molecular background of isolated COX deficiency in our patients. In addition of simple characterization of molecular background and optimalization of methods for routine diagnostics, we were able to study the impact of several mutations in nuclear genes for COX assembly factors on the biochemical, structural and histochemical level in affected tissues.
|
|
Hemoprotein Nitric Oxide Synthase in Aplysia Californica
Buganová, Michaela ; Martásek, Pavel (advisor) ; Pelouch, Václav (referee) ; Druga, Rastislav (referee)
Nitric oxide (NO) plays a crucial role in neuronal signaling in a variety of eukaryotic and prokaryotic organisms. Nitric oxide synthases (NOS) are heme-containing monooxygenases that catalyze the oxygen dependent oxidation of L-arginine to NO and L-citrulline. The NO produced by NOS activity is a gaseous molecule that diffuses easily through membranes and acts inter or intracellularly. NO activates metal-containing enzymes, including soluble guanylate-cyclase (sGC) that increase levels of the messenger molecule cyclic 3,5-guanosine monophosphate (cGMP) (1, 2) which in turn mediate various pathophysiological or physiological functions in neurons. Nevertheless, many aspects of nitrergic neurons and NO function in the central nervous system (CNS) are unclear. The aim of research described in this thesis was to characterize neuronal NOS, proteins metabolically linked to NOS and NO signaling pathways in the CNS of Aplysia cali/ornica (Aplysia), a popular experimental model in cellular and system neuroscience. The biochemical characteristics of Aplysia NOS (AcNOSj described here revealed its calcium-/calmodulin-(Ca/CaM) and NADPH dependence. A representative set of inhibitors for mammalian NOS isoforms also suppressed NOS activity in Aplysia Polyclonal anti-rat nNOS antibodies hybridized with a putative purified...
|
|
Effect of equilibration and freezing parameters on bull sperm motility after thawing
Beránková, Monika ; Stádník, Luděk (advisor) ; Zuzana, Zuzana (referee)
The hypothesis assume the existence differences in bulls spermatozoa viability after thawing, characterized by higher values of motility and higher proportion of live spermatozoa, in case of prolonged equilibration and lower temperature freezing gradient. The aim of this thesis was to find the influence of different equilibration and freezing curve conditions to on spermatozoa viability after thawing. The theoretical part of the thesis summarizes available information about the bulls semen parameters, methods for its evaluation and especially the processing and long-term preservation.
Within the case study part of the thesis, during the years 2013-2015 the semen of the pre-selected group of 5 breeding bulls was being collected at the Sires insemination center. The group was uniform as to age, breed and housing and treatment conditions. After initial assessment the collected semen was divided into parts, which were then processed in different wals. A part of straws was processed by standard protocol, the other part was processed using different length of equilibration and freezing curve.
First part of the straws was equilibrated in a standard protocol for 120 minutes; subsequently half of the equilibrated straws was standardly frozen by the Direct Freezing method based on 3-phase freezing curve while the other half was frozen using the different 2-phase freezing curve. Second part of the straws was initially equilibrated for 240 minutes with the subsequent freezing, similarly as in the previous case, used 3-phase and 2-phase freezing curve. Frozen straws were then stored in liquid nitrogen container at -196 °C.
Spermatozoa motility was evaluated using CASA. The semen was further evaluated on the proportion of live and dead spermatozoa, using coloration of Eosin and Nigrosine, and on the sensitivity of the spermatozoa membranes, using HOS test, which detects the reaction of spermatozoa tail membrane to hypoosmotic conditions.
The obtained results indicated higher progressive spermatozoa motility (+5.57 %) and higher percentage of live spermatozoa (+4.47 %) following the application of the prolonged equilibration (240 min). When applying the length of equilibration 240 minutes, the values of total spermatozoa motility and HOS test were higher as well, though the differences were not statistically significant. When evaluate the effect of using different freezing curves on the parameters of the spermatozoa after thawing, higher values (average + 1.6 %) at almost all of these parameters using 3-phase freezing curve were found, the diferrences were not statistically significant.
Statistically significant differences among the bulls (P < 0.01) were found in total as well as progressive spermatozoa motility, in the proportion of living spermatozoa and in value of the HOS test - the individuality among the bulls is therefore evident. While evaluating the characteristics of spermatozoa movement by CASA, statistically significant differences among the bulls were confirmed at VCL, VSL, VAP, LIN, STR, WOB (P < 0.01), also between the diferrent length of equilibration (P < 0.01), but were not statistically significant when using diferrent freezing curves.
|
|
Characterization of barbiturates and benzodiazepines-human serum protein interactions by cappilary electrophoresis
Studničková, Kamila ; Wsól, Vladimír (advisor) ; Pospíšilová, Marie (referee)
An important topic in the drug discovery and development process is the role of drug binding to plasma proteins. In this diploma thesis the characterization of the interaction between benzodiazepines and barbiturates towards human serum albumin and α-1-acid glycoprotein under physiological conditions by capillary electrophoresis-frontal analysis is presented. Furthermore, the binding of these drugs to all plasma proteins is evaluated by using ultrafiltration and capillary electrophoresis. The results indicate that the hydrophobic character of compounds seems to be the key factor on the interaction between these drugs towards proteins. In fact, hydrophobic basic drugs (benzodiazepines) bind in great extension to HSA, while less hydrophobic acid drugs (barbiturates) present lower interactions with proteins and bind especially to AGP.
|
|
Treatment of diabetes mellitus
Hrouzková, Šárka ; Tilšer, Ivan (advisor) ; Hrdina, Radomír (referee)
DM is metabolic disorder characterized with hyperglycaemia at absolute shortage of endogenous insulin (DM1T), or at its relative shortage (DM2T). DM1T is a symptom of B-cell destruction in islets of pancreas Langerhans that occurs mostly on autoimmune basis. Lack of insulin results in disorder of glucose use and in increased catabolism of fat and proteins. The result of this disorder is hyperglycaemia, glycosuria, polyuria, thirst, eventually ketoacidotic coma. DM2T which prevalence is continuously increasing represents significant medical, social and economic problem. DM2T develops due to insulin resistance and later due to insufficient insulin secretion after each meal. Insulin resistance mostly develops in connection with obesity. In the course of disorder the defect of insulin secretion gradually worsens and micro and macro vascular complications develops that are the most frequent cause of death of patient suffering from the disorder. Diabetes treatment is of complicated and complex nature that has to be based on simultaneous intensive therapy, prevention, collaboration with patient, education, individual adjustment of treatment and therapeutic goals. The Czech Republic belongs to a few countries where diabetology as an independent branch of medicine exists. The goal of treatment is to ensure...
|
guest ::
