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Cortical electrical stimulation and pain
Rusina, Robert ; Rokyta, Richard (advisor) ; Haninec, Pavel (referee) ; Paleček, Jiří (referee)
The aim of the study was to examine effects of sensorimotor cortex stimulation on pain in animal. A behavioral model investigated pain thresholds in deafferentated rats depending on cortex stimulation and two neurophysiological models studied different components of the jaw opening reflex (JOR) and tooth pulp evoked potentials (TPEPs) following cortical stimulation. The behavioral model used 18 deafferentated (dorsal root rhizotomy) rats and 14 controls. Pain thresholds were measured before and after cortical stimulation using plantar test and tail-flick latencies. In the neurophysiological model, rats were implanted with tooth pulp, cerebral cortex, and digastric muscle electrodes. 15 animals were divided into three groups, receiving 60 Hz, 40 Hz and no cortical stimulation, respectively. TPEPs were recorded before, one, three and fi ve hours after continuous stimulation. 10 other rats were submitted to recordings after a single tooth pulp stimulation, while in 5 more rats we administrated conditioning and test stimulation. TPEPs and digastric EMG were simultaneously recorded. A multiresolution denoising method was used for signal processing. Our results show a similar effect of the stimulation in man and experimental animals despite the differences in the organization of the cerebral cortex. Our results...
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Molecular factors of artherogenesis in chronic kidney disease with experimental model of chronic inhibition of Nitric Oxide Synthesis and its Effect of HMG-CoA reductase inhibitoin
Lecian, Dalibor ; Matouš, Bohuslav (advisor) ; Tesař, Vladimír (referee) ; Zadražil, Josef (referee)
The experimental part of the thesis covered the markers involved in nitric oxide deficiency model in rats. The analogue of arginine N-nitro- L -arginine-methyl ester ( L -NAME) was used. L -NAME administration resulted in sustained elevation of BP, decreased GFR, and in higher albuminuria compared to control animals. The administration of hypolipidemic drug - atorvastatin to Wistar rats was studied. The statin used caused lower albuminuria and prevented decreases in GFR in L -NAME rats without having an impact on hypertension. CAV1 protein expression was similar in all groups of rats. In contrast, VEGF expression and RhoA activity was increased in L -NAME-treated animals, and normalized with coadministration of atorvastatin. Clinical part of the thesis focused on the analysis of markers of oxidative stress in obese and non-obese patients with CKD and after kidney transplantation. The patients with CKD had elevated serum concentrations of AGEs, sRAGE and ADMA with the maximal concentrations of AGEs in the patients on hemodialysis. The successful kidney transplantation decreases the AGEs, sRAGE and ADMA concentrations but not to normal value according to general population. Patients with obesity exhibited lower concentrations of sRAGE in CKD as a protective factor for cardiovascular disease. Concentrations of...
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Effects of Spirulina platensis on the expression of endothelial NO synthase in apoE-deficient mouse model of atherosclerosis
Pešlová, Monika ; Nachtigal, Petr (advisor) ; Kopecký, Martin (referee)
1. ABSTRACT Spirulina platensis is a blue-green microalga belonging to the cyanobacteria family. Microalga is a large source of proteins and good fatty acid and other nutritional elements, for example iron, calcium, chromium, lithium, selenium. It also contains natural dyes chlorophyll, phycocyanin. Its antioxidant and anti-inflammatory were described. The aim of this thesis was to determine potencial hypolipidemic effects and potential effects on endothelium of Spirulina platensis in apoE-deficient mice. ApoE-deficient mice were fed standard diet for 2 weeks. At the age of 8 weeks the control group of animals were fed with the western type diet, which contained 21% fat and 0,15% cholesterol for 8 weeks. The same atherogenic diet was used in Spirulina platensis group, where Spirulina platensis was added to the atherogenic diet at the dosage of 20 mg per day. The biochemical analysis of lipid spectrum was done, area of atherosclerotic lesions was determined and imunohistochemical and stereological analysis of eNOS expression was performed as well. The results of this thesis showed positive effects of Spirulina platensis on cholesterol levels, VLDL and LDL cholesterol. Moreover stereological analysis of imunohischemical staining revealed that, that endotelial expression eNOS was significantly increased by...
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Monitoring the effect of atorvastatin on the expression of endoglin / / and endothelial NO synthase (eNOS) in mouse model of atherosclerosis
Uhlárová, Iva ; Nachtigal, Petr (advisor) ; Štaud, František (referee)
Atherosclerosis, or sclerosis of arteries, is a degenerative disease of arteries. Sometimes it is called "the disease of 20th century". ApoE/LDL - receptor double knockout mice represent a new animal model for study of atherogenesis, which is characterized by severe hyperlipidaemia and atherosclerosis. Statins (or competitive inhibitors 3-hydroxyl-3-methyl-glutaryl-coenzym A-reductase) currently belong to the most efficient and the most useful hypolipidemic drugs for all over the world. They decrease mainly levels of total cholesterol and LDL cholesterol. The aim of this thesis was to describe the expression of endoglin in atherosclerotic plaques in apoE/LDL-receptor deficient mice. Moreover we wanted to determine endoglin colocalization with eNOS and the effect of atorvastatin treatment on the expression of both molecules. ApoE/LDLR-deficient mice on were subdivided into 2 groups. The control group of animals was fed with the western type diet. The same atherogenic diet was used in ATV group, where atorvastatin was added to the atherogenic diet at the dosage of 100 mg/kg per day. The results of this thesis confirmed the expression of endoglin in atherosclerotic lesions in ApoE/LDLR-deficient mice. The expression of endoglin was located on the aortic vascular endothelium and in other smaller...
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Calculus Bovis effect on blood lipids and the expression of endothelial NO synthesis / / in ApoE-deficient mouse model of atherosclerosis
Studená, Romana ; Nachtigal, Petr (advisor) ; Trejtnar, František (referee)
Calculus Bovis is one of the most precious and commonly-used medicinal materials in China. It is consists predominantly bilirubin and bile acids. In this thesis we focused on possible effects of Calculus Bovis on lipids and atherogenesis in apoE-deficient mice. ApoE-deficient mice were fed standard diet for 2 weeks. At the age of 8 weeks the control group of animals were fed with the western type diet, which contained 21% fat and 0,15% cholesterol for 8 weeks. The same atherogenic diet was used in Calculus Bovis group, where Calculus Bovis was added to the atherogenic diet at the dosage of 50 mg/kg b.wt. Calculus Bovis (Artificial bezoar, Biopharma, Australia) per day. The biochemical analysis of lipid spectrum was done, area of atherosclerotic lesions was determined and imunohistochemical and stereological analysis of eNOS expression was performed as well. Calculus bovis treatment did not affect blood lipids, total cholesterol, VLDL, LDL, HDL and TAG when compared with control mice. Moreover Calculus bovis treatment did not decrease either atherosclerotic plaque area or endothelial expression of eNOS. The failure of Calculus Bovis treatment in this pilot study could be probably related to insufficient dose of Calculus Bovis.
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Information content in vocalization of guinea pigs (Cavia porcellus): individual distinction and recognition of predators
Baklová, Aneta ; Baranyiová, Eva (advisor) ; František, František (referee)
Guinea pigs represent domesticated precocious rodents which became common pets. From the first day after birth they are fully vocal. This thesis was devoted
to the vocalization of pups. The aim of the thesis was to 1) determine the age when the vocal individuality in whistle sound is demonstrable; 2) test possible ultrasonic signals emitted by young guinea pigs; 3) study the antipredator reactions and alarm calls to aerial (bird of prey) and terrestrial (dog) predators and human (control test).
A total of 16 guinea pig pups were tested for vocal individuality, 28 pups
for ultrasonic vocalization and 27 adolescents for predator recognition. When testing vocal ontogeny during the first 9 days after birth, I observed changes in temporal, frequency and parameter of intensity . When I test vocal individuality by cross-validated discriminant function analysis (DFA) based on ten acoustic parameters, calls were classified to correct animals with following success: day 1 = 71.9%, day 3 = 58.8%, day 5 = 53.10%, day 7 = 50.60% and day 9 = 63.10%. The highest frequency in whistle was 30.03 kHz. In predator discrimination reactions as freezing, fleeing and vigilance were observed. In the presence of dog, guinea pigs reacted for the longest time and most frequently by freezing. When confronted with a bird of prey, I observed for a longest time and most often fleeing and then freezing. When exposed to a human, guinea pigs showed mostly vigilance. Almost no vocalization was observed except for two events
of alarm calls - drrr as in the presence of dog and chirrup as reaction to bird of prey.
The following conclusions can be drawn from the presented results: 1) vocal individuality of guinea pigs is demonstrable immediately after birth and the rate
of individually different vocal parameters changes with age; 2) guinea pigs are able
to produce sound up to 30 kHz, i.e. within the ultrasound range, but signals of high frequencies are not crucial for their communication; 3) guinea pigs discriminate between terrestrial and aerial predators, but they emit alarm calls rather rarely.
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Emissions of greenhouse gases (N2O and CO2) from pasture soil as a result of activity of soil microbial community - preliminary results
Šimek, Miloslav ; Hynšt, Jaroslav ; Brůček, Petr ; Čuhel, Jiří
Cattle overwintering area was shown an important point source of N2O and CO2. The emissions of N2O were mostly directly related to the rate of animal impact as the greatest fluxes of N2O and the highest total cumulative N2O emissions as well were found at the most impacted site. The fluxes of N2O were however very variable in time and most of N2O was emitted during several short events in spring or in winter. Laboratory experiments suggested that total amount of nitrogen escaping from soil from impacted sites is much greater than indicated by field measurements of N2O fluxes and molecular nitrogen (N2) was shown as the main nitrogen gas. It is hypothesized that soil pH is a crucial soil factor controlling the mole fraction of N2O: under slightly alkaline conditions in severely impacted soils, denitrification is completed and most of nitrogen escapes from the soil as N2; under more acidic conditions, however, the activity of nitrous oxide reductase is partly reduced.
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CD46 and β1integrin interaction in mouse sperm head
Šebková, Nataša ; Frolíková, Michaela ; Dvořáková-Hortová, Kateřina
CD46 protein plays an important role during fertilization and its role is associated with acrosome stability. CD46 is probably involved in signalling pathways triggering the acrosome reaction. Integrins interact with many cytoskeletal proteins such as actin, therefore changes in the actin cytoskeleton before and after AR may lead to changes in the association and localization of CD46 and β1integrin. Our aim was to monitor mutual CD46 and β1integrin interaction detected by the proximity ligation assay. It generates a localized signal in a form of spots revealing the exact position of the recognition event. Proteins interaction was study in freshly released sperm and sperm during the acrosome reaction, during which there is a gradual relocation of these proteins towards the equatorial segment and the whole sperm head. Proteins α and β tubulin were used as a positive control, α tubulin and β1 integrin as a negative control. In situ PLA showed a distinct spotted signal indicating the mutual interaction of CD46 and β1integrin. A positive response was demonstrated not only in freshly released sperm but also in sperm during the acrosome reaction. Freshly released sperm were distinctively labelled in the acrosome region and the neck, similarly to the positive control. Sperm during the acrosome reaction showed the signal across the whole sperm head region. No signal or sporadic nonspecific staining was detected in the case of the negative control. In summary, our results deliver new information that proteins CD46 and β1 integrin interact with each other. These results suppose the theory that β1 integrin can mediate a connection between CD46 and sperm cytoskeleton thereby molecules of signalling pathways leading to activation of the acrosome reaction.
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Angiogenesis in cerebral ischemia border zone
Nulíčková, Radka ; Otáhal, Jakub (advisor) ; Brime, Tufikameni (referee)
Title of work: Angiogenesis in the border zones of cerebral ischemia The aims of work: The aim of this diploma thesis was to determinate the parameter of the angiogenesis after focal brain ischemia induced by photothrombosis in P7 rat pups. This thesis should answer the question, if there is some difference in vascular density according to the distance from the lesion and if there is some diference between the vascular density in normal brain and the brain with the lesion. Materials and Methods: Cerebral ischemia was induced in P7 rat pups with photothombic method. Intravenous administration of bengal rose dye was followed by illumination of beam of green laser over the cortex for 5 minutes at 0,5s on/off cycles. Bengal rose dye is potent photosensitive substance and the ilumination resulted in agregation of trombocytes and thrombosis. Cerebral ischemia evolved due to the thrombosis. After two months, the animals were overdosed with urethane and transcardially perfused. Then thein brain were cryoprotected and then cut into 50µm slices and prepaired for histological staining. The stained slices were the investigated under mikroskope. The microvessels in the cortex were counted and the density was estimated by stereological method. Results: Two brain with ischemic lesion and two control brain were...
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