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The effect of morphine withdrawal on the adenylyl cylclase signaling system in rat brain
Nováková, Daniela ; Novotný, Jiří (advisor) ; Rudajev, Vladimír (referee)
The aim of this bachelor thesis is to study the effect of morphine withdrawal on the adenylyl cyclase signal system in the rat brain. Theoretical part of this thesis summarizes basic information known about opioids and its leading representative morphine, molecular mechanisms of its effect on the organism and yet determined knowledge about effects of morphine addiction and subsequent withdrawal on the organism and above all on the central nervous system. There is still much unknown about exact mechanism of development of morphine addiction and withdrawal. This thesis also summarizes briefly the Western blot method used in analysing the effect of morphine withdrawal on selected proteins of the adenylyl cyclase signal system in the rat brain. Experimental part of this thesis is based on the detection of expression of key components of the adenylyl cyclase pathway in selected regions of the rat brain after long-term administration and subsequent withdrawal of increasing doses of morphine. Results of this study did not reveal statistically significant differences in the expression of adenylyl cyclase signaling system components between the withdrawal and the corresponding control groups of animals, confirming high ability of the organism to withstand morphine administration and to return to...
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Sexual dimorphism of rat gut microbiota composition and intestinal immunity
Kalousová, Pavla ; Kovařík, Miroslav (advisor) ; Jílek, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Pavla Kalousová Supervisor: PharmDr. Miroslav Kovařík, Ph.D. Title: Sexual dimorphism of rat gut microbiota composition and intestinal immunity Background and Aim: Many factors can influence the composition of gut microbiota and the immune system. It is well-known that one of those factors is sex. This sexual dimorphism can lead to a specifically adjusted treatment of diseases for different gender and nutritional interventions. This study focuses on analysing sexual differences in gut microbiota and intestinal immunity in adult rats. Methods: Caecal content from 12-week-old female and male Wistar rats were collected and analysed by DNA-sequencing technique to characterize microbiota composition. ELISA test was performed to quantify the concentration of IgA in faeces and gut wash. Flow cytometry determined the concentration of IgA-coating bacteria in faecal samples. Results: Metagenomic analysis revealed that female gender presents 1 phylum, 4 families, 13 genera, and 13 species which are not present in male rats. Only one male-specific colonization was observed at the species level. Quantitative analysis showed a higher proportion of Firmicutes phylum in males which was associated with...
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Analysis and Detection of RAT Malware
Sidor, Samuel ; Frolka, Jakub (referee) ; Hajný, Jan (advisor)
Goal of this bachelor’s thesis is studying problematics of various types of malware with specific focus on RAT (Remote Access Trojan) category. This thesis will also acquaint reader with static and dynamic binary analysis and terms like reverse engineering, sandboxing, decompilation, etc. Then chosen malware families will be analysed and for these families detection rules in YARA language will be created. Except this, reader will be acquainted also with protection against RAT malware and finally data acquired from detail analysis will be evaluated.
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Evidence securing in virtual environment
Havlová, Miluše ; Malina, Lukáš (referee) ; Martinásek, Zdeněk (advisor)
The bachelor thesis deals with evidence securing in virtual environment. The main goal is to suggest suitable virtual environment for evidence securing which can be used in prosecution. As the next the tools that are suitable for safe securing of evidence are described. With the help of selected tools the sample of malware is secured and then the sample is analyzed and described. The suggested environment is used to evidence gathering within the frame of simulated security incident.
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The in vitro effects of selected fenolic substances on isolated rat vessels
Rajtmajerová, Iveta ; Pourová, Jana (advisor) ; Hrdina, Radomír (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Iveta Rajtmajerová Supervisor: PharmDr. Jana Pourová, Ph.D. Title of diploma thesis: The in vitro effects of selected fenolic substances on isolated rat vessels Silymarin flavonolignans are endowed with various aspects. Recently their hepatoprotective effect has been discussed widely as well as their positive influence on the cardiovascular system. This diploma thesis is concerned with the vasodilatory effects of silybin-A, silybin-B, silybin-A + B and isosilybin-A on rat aorta. The aim of this study was to test the vasodilatory effect of selected substances. The dependence of aortic relaxation on the increasing concentration of tested substances was measured. Isolated Wistar breed rat aorta was used. DRC curves were created and EC50 values were assessed from the obtained values of vessel tension. The results were evaluated subsequently. The vasodilator potential of silybin-A and isosilybin-A is comparable; hence the position of the substituents may be irrelevant in having any impact on relaxation of the vessels. Different vasodilatory effects of silybin-A and silybin-B have been observed thus, the relaxation could be stereoselective. The testing of the equimolar mixture of silybin-A and...
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Bezpečná anestezie potkana pro náročné výkony
NOVOTNÁ, Nikola
Rat is a significant laboratory animal that is used especially in pharmacology, experimantal surgery, onkology and in reproduction and nutrition experiments. Therefore, it is very important to know the dosage of anestetics even for longer an more demanding operations. This bachelor thesis deals mainly with optimimization of anesthetic doses for demanding procedures and comparison of effects of individual anestecics on rat physiology. In the experiment, rats were divided into five experimental groups. The first group was given a combination of ketamine and xylazine totaling 3 times, where the first dose was for ketamine 100 mg/kg and 10 mg/kg for xylazine; the second and third doses were dosedfor ketamine 50 mg/kg and 0.5 mg/kg for xylazine. The second group was also administered 3 times with propofol, medetomidine and nalbuphine, where the first dose was for propofol 100 mg/kg, 0.1 mg/kg for medetomidine and 10 mg/kg for nalbufine; the subsequent anesthetic dosing was 50 % of the initiation dose. The third group was given a combination of propofol, medetomidine and nalbuphine again 3 times, where the first dose was 100 mg/kg for propofol, 0.1 mg/kg for medetomidine and 10 mg/kg for nalbuphine; the subsequent anesthetic dosing was 25 % of the initiation dose. The fourth group was given a combination of ketamine and xylazine only once, where ketamine was 100 mg/kg and 10 mg/kg for xylazine. The last fifth group was given a combination of medetomidine and nalbuphine only once, where the dosage was 100 mg/kg for propofol, 0.1 mg/kg for medetomidine and 10 mg/kg for nalbufine. After inhalation of isoflurane, the rats were given intraperitoneal anesthetics. The results show that the experimental groups used, long-term anesthesia (3 doses of anesthetics) was most, beneficial for rats using a combination of propofol (100 mg/kg), medetomidine (0.1 mg/kg) and nalbuphine (0.1 mg/kg) kg), where the subsequent doses were 25 % of the initial dose. Furthermore, we found that ketamine and xylazine anesthesia is insufficient for long-term surgery. The prolongation of the surgical phase of anesthesia by repeated administration of a reduced dose of anesthetics is possible, but this anesthesia is characterized by a more frequent occurrence of complications and therefore generally not recommended.
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Cardioprotective effect of adaptations to hypoxia and cold: Connexin 43 and hexokinase 2
Kohútová, Jana ; Žurmanová, Jitka (advisor) ; Radošinská, Jana (referee) ; Barančík, Miroslav (referee)
Ventricular arrhythmias are the main cause of death worldwide. An increased incidence of arrhythmias in the heart of mammals is accompanied by a remodeling of the cellular distribution gap between the channels of mainly connexin 43 (Cx43). Recently has been demonstrated significant effect of mitochondria and their association with arrhythmogenesis. Various pathological conditions alter the expression and / or distribution of Cx43, depending on the phosphorylation status. but also on altering the association of hexokinase with mitochondria, which reduces the likelihood of apoptosis activation. Adaptation to intermittent hypobaric hypoxia potentiates endogenous pathways reducing the incidence of ventricular arteries, whereas continuous normobaric hypoxia does not have this effect. Another studied model is cold acclimatization, which has been known for several decades by known effects on human health. However, the heart study of these models in relation to Cx43 is missing. Our goal was to determine the expression of Cx43, phosphorylated Cx43 (p-Cx43) and hexokinase (HK1, HK2) and their distribution in cardiomyocytes. In addition, the expression of Cx43 upstream kinases, protein kinase A, protein kinase G, casein kinase 1 in normoxic and hypoxic left ventricles of rats, along with Cx43 distribution...
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Imunomodulační účinky extraktů z helminta na střevní buněčnou linii potkaního modelu
LEVÁ, Jana
In this study, we examined the immunomodulatory effect of excretory/secretory products, crude adult extracts and crude larvae extracts from Hymenolepis diminuta on the intestinal epithelilal cell line from a rat. For determination of the immunomodulation effect of all H. diminuta extracts was used relative gene expression of TNFa, IL-17re and IL-33 from epithelial cells and it was tested using real-time PCR. Our result showed that excretory/secretory products had the strongest antiinflammatory effect on the epithelial cells. We assume that crude adult extracts play an important role in increase of gene expression of IL-33 and also in the immunomodulatory ability of H. diminuta in the host organism.
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Analysis of oxidative stress markers in rat brain: the effect of maternal separation
Pallag, Gergely ; Novotný, Jiří (advisor) ; Vodička, Martin (referee)
Adverse events that cause stress during the early stages of life may alter the normal development of the brain and neuroendocrine system and increase the vulnerability of the individual to various disorders. Chronic stress and subsequent releasing of stress mediators can lead to oxidative stress and cell damage. The first aim of this work was to determine selected oxidative stress markers in the cerebral cortex, hippocampus, and cerebellum after the exposure of rats to early life stress. To model the stressful situation, we used maternal separation of the offspring for three hours a day during the first three weeks of life. We choose reduced glutathione, protein carbonyls, lipid peroxides and hydroperoxides as typical markers. These markers were determined in the brains of rats aged 22 days. Any significant changes were found in the levels of the studied markers after maternal separation. Damage to brain cells may also be reflected in behavior. Studies of numerous neuropsychiatric and neurodegenerative diseases have indicated that oxidative stress is a promising candidate for inducing changes at the cellular level. The second aim of this work was to monitor the behavior of rats by the light/dark box test after maternal separation along with administration of N-acetylcysteine (NAC), a drug with...
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Inter-individual differences in behaviour of laboratory rats
Rudolfová, Veronika ; Nekovářová, Tereza (advisor) ; Sedláček, František (referee)
Number of studies report that even when experimental animals are subjected to the exact same conditions, they differ in their behaviour. If these differences were stable in time and across several experimental procedures, we could talk about personality. This diploma thesis studies inter-individual differences in behaviour of laboratory rats (Long Evans strain) in a series of experiments conducted in early ontogeny and in adult age. Apart from analysing inter-individual differences in behaviour and personality of experimental animals, this thesis has two main aims. The first aim is assessing stability of inter-individual differences in behaviour throughout ontogeny. The second aim is to explore possible link between inter-individual differences in behaviour and performance in cognitive tests. We confirmed the existence of inter-individual difference in behaviour in laboratory rats. In this thesis we were, however, not able to assess personality of experimental animals. The differences in behaviour were best described by behaviour in Open field test and Elevated plus maze test. Our results also show marked differences between successive trials of these experiments. We also report that performance in Active allothetic place avoidance is not linked to performance in Morris water maze, even though both...
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