|
| |
|
|
Circadian system as a modulator of neuroinflammation
Kotková, Eliška ; Spišská, Veronika (advisor) ; Dočkal, Tereza (referee)
The circadian system is involved in the regulation of biological rhythms in physiological, behavioural and immune processes. These rhythms can be found in the central nervous system, including the blood-brain barrier, astrocytes, microglia, and the pineal gland, which produces the hormone melatonin. Neuroinflammation is a complex response of the central nervous system to inflammatory stimuli by rhythmic expression of pro-inflammatory and anti-inflammatory mediators or by rhythmic regulation of immune system cells. Studies have examined the influence of genes and proteins of the circadian system, suprachiasmatic nuclei, melatonin, and glial cell rhythms on neuroinflammation. Lipopolysaccharide was used to induce neuroinflammation in these studies. Based on these studies, the effect of melatonin on mikroglia and endothelial cells, and the responses of suprachiasmatic nuclei was evaluted as the most important circadian modulator of neuroinflammation. This thesis describes the basic principles of the circadian system and neuroinflammation, with the last section presenting the modulation of neuroinflammation by the circadian system. Keywords: astrocytes, blood-brain barrier, circadian system, cytokines, immune system, melatonin, microglia, neuroinflammation, suprachiasmatic nuclei
|
|
|
Simulation of Perfusometric Data
Hývlová, Denisa ; Mézl, Martin (referee) ; Jiřík, Radovan (advisor)
This diploma thesis focused on the simulation of perfusion MRI data. The introduction describes standard perfusion imaging methods, DCE and DSC, and the quantitative analysis of perfusion. In the following section, simulation of the perfusion MRI data and the structure of the provided simulation software are described. This software was reworked, and new methods and parameters were implemented. The key product of this work is a simulation study of focused ultrasound-induced blood-brain barrier opening, comparing the pharmacokinetic models used in this application and the accuracy of their permeability estimates. The results of the simulation study were compared with real-dataset results.
|
|
|
Gut microbiome and autoimmune mechanisms in patients with type 1 narcolepsy
Ježková, Janet ; Roubalová, Radka (advisor) ; Hrdý, Jiří (referee)
Narcolepsy Type 1 (NT1) is a chronic neurological disease characterized by the presence of cataplexy and excessive daytime sleepiness. NT1 probably arises from autoimmune destruction of orexinergic neurons in the hypothalamus. These neurons are the only known producers of the neuropeptides orexin A and B, which are involved in the regulation of sleep and wakefulness. NT1 is often accompanied by comorbidities such as obesity, type 2 diabetes, hormonal disorders, depression, and anxiety. Gut microbiota affects the quality of sleep by the production of various metabolites. It is considered that it may be involved in the pathogenesis of NT1 or in the development of related comorbidities. In our study, we analyzed the gut microbiota composition of 41 NT1 patients and 32 healthy controls using next-generation sequencing. The diversity of patients' gut microbiota did not differ significantly from healthy controls. In patients, we revealed a positive correlation between the abundance of the bacterial family Coriobacteriaceae, BMI, and the disease duration. Besides, we observed a negative correlation between the Coriobacteriaceae family and cholesterol levels, suggesting that these bacteria are involved in host lipid metabolism. Compared to healthy controls, a higher abundance of bacteria from the families...
|
|
|
Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method
Šinaľová, Lucia ; Hofman, Jakub (advisor) ; Jansová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucia Šinaľová Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method This master thesis focuses on the prediction of tacrine derivatives permeability through blood- brain barrier and their cytotoxicity. We studied tacrine-benzothiazole and tacrine- thiaquinazoline derivates as potential drugs for treatment of Alzheimer's disease. The reason of testing new tacrine derivates was effort to find substances with same effect and lower toxicity, for which was the tacrine discarded from clinical practice. The probable permeability of the studied substances was determined in vitro by the PAMPA method. Based on the permeation coeficient values we have identified that tacrine-benzothiazole derivates A-D (Pe= 8,31- 16,9×10-6 cm/s) and tacrine-thiaquinazoline derivates 1, 3, 4 (Pe= 8,59-14,9×10-6 cm/s) were permeable through blood-brain barrier, tacrine-thiaquinazoline derivate 2 (Pe= 3,79×10-6 cm/s) had uncertain permeability and tacrine-thiaquinazoline derivate 5 (Pe= 2,0×10-6 cm/s) was not permeable. In the pre-clinical evaluation of...
|
|
|
Study of exosomes as drug delivery system in therapy of glioblastoma
Tomášková, Lucia ; Šimůnek, Tomáš (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lucia Tomášková Supervisor: prof. PharmDr. Tomáš Šimůnek, Ph.D. Title of diploma thesis: Study of exosomes as a drug delivery system in the treatment of glioblastoma Central nervous system disorders are among the most serious diseases affecting humans. They affect not only the patient's life, but also his/her surroundings. Therefore, their therapy, whether at the level of complete cure or alleviation of accompanying symptoms, is a challenge for scientific research. In our research, we focused on glioblastoma multiforme, a brain cancer not yet treatable. The main drawback in therapy is overcoming the blood-brain barrier. Exosomes, such as the body's natural nano-vesicles, have been shown to be a suitable system for delivering drugs to brain tissue. Our research has shown that by a suitable method we are able to obtain sufficient quality exosomes from macrophage and fill them very efficiently with antitumor agents paclitaxel, doxorubicin and temozolomide, while the delivered substances show higher efficacy and fewer side effects than the free form.
|
|
|
Acute neuroinflammatory reaction after focal cerebral ischemia
Ambrož, Ondřej ; Otáhal, Jakub (advisor) ; Konopková, Renata (referee)
Title: Acute neuroinflammatory reaction after focal cerebral ischemia Aim: The aim of this thesis is to evaluate neuroinflammatory response after focal cerebral cortical ischemia. Also, familiarizing with the method of displaying damage of blood brain barriers, neurons and the possibility of detection of microglia cells as a marker of acute neuroinflammatory processes. Methodology: This is an experimental study. We brought about cortical cerebral ischemia in rats using an application of photosensitive dye "bengal red," and a green laser. Two animals were were given the additional application of "Evans blue" in order to visualize the defects of the blood brain barrier. The animals were returned to their cage for the time needed before they were induced terminal anesthesia. This was followed by the process of brain perfusion, slicing the brain in sections 50 µm thick and then applied these sections onto slides. Sections with applied EB were immediately analyzed under the microscope. Sections to illustrate neuronal death were immunohistochemically stained via the Nissl method. Sections visualizing microglial activity were stained using CD11b antibody. Results: Following the induction of focal ischemia there occured brain tissue damage. In the vicinity of lesion there is degeneration of neurons and...
|
|
|
Preparation of oxim reactivators loaded PLGA nanoparticles
Veliu, Mimoza ; Šnejdrová, Eva (advisor) ; Smékalová, Monika (referee)
5 2 ABSTRACT Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Technology Consultant: PharmDr. Eva Šnejdrová Ph.D. Student: Mimoza Veliu Title of Thesis: Preparation of oxim reactivators loaded PLGA nanoparticles In theoretical part the main attention is paid to polymeric nanoparticles (NP) for brain targeting. The drug properties and methods of preparation of the nanoparticles with hydrophilic drugs are presented. The part is devoted to physico-chemical properties that affect the permeability of these substances across biological barriers to the target site. The polymers used for NP preparation for brain targeting are focused. The experimental part deals with the nanoparticle formulation. Nanoparticles were prepared by double emulsion method. The method of preparation was chosen based on the drug solubility. In the W/O/W system oil phase was formed by linear copolymers of poly(lactic-co-glycolic) acid (PLGA) and its derivatives branched on polyacrylic acid (A2) or tripentaerythritol (T3). Poloxamer 407 and dimethyldidodecylammonium bromide (DDAB) were chosen for NP stabilization. The effect of the polymers on particle size was studied. The effect the polymers used, and the formulation factors on the size, polydispersity and stability of NP were studied. The concentration...
|
|
|
Study of exosomes as drug delivery system in therapy of glioblastoma
Tomášková, Lucia ; Šimůnek, Tomáš (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lucia Tomášková Supervisor: prof. PharmDr. Tomáš Šimůnek, Ph.D. Title of diploma thesis: Study of exosomes as a drug delivery system in the treatment of glioblastoma Central nervous system disorders are among the most serious diseases affecting humans. They affect not only the patient's life, but also his/her surroundings. Therefore, their therapy, whether at the level of complete cure or alleviation of accompanying symptoms, is a challenge for scientific research. In our research, we focused on glioblastoma multiforme, a brain cancer not yet treatable. The main drawback in therapy is overcoming the blood-brain barrier. Exosomes, such as the body's natural nano-vesicles, have been shown to be a suitable system for delivering drugs to brain tissue. Our research has shown that by a suitable method we are able to obtain sufficient quality exosomes from macrophage and fill them very efficiently with antitumor agents paclitaxel, doxorubicin and temozolomide, while the delivered substances show higher efficacy and fewer side effects than the free form.
|
|
|
In vitro Methods for the Prediction of Blood Brain Barrier Penetration
Zálešáková, Helena ; Vopršalová, Marie (advisor) ; Čečková, Martina (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Helena Zálešáková Supervisor: PharmDr. Marie Vopršálová, CSc. Title of diploma thesis: In vitro Methods for the Prediction of Blood Brain Barrier Penetration This thesis deals with the correlation between two in vitro models simulating the blood- brain barrier (HEB, hematoencephalic barrier) and their comparison in terms of practical use. These are the PAMPA (Parallel Artificial Membrane Permeability Assay) method and the MDCK (Madin-Darby Canine Kidney) cell line, which are models for potential central nervous system (CNS) penetration screening. Within this work, a set of sixteen standard drugs were measured. The procedure was similar in both methods in order to obtain information on the amount of test substances passing through the membrane from the donor portion of the plate to the acceptor. The concentration in the donor portion was measured by UV-VIS spectrophotometry. The main difference between these methods is the membrane through which the substances penetrate. In the case of PAMPA, a lipid solution that has been isolated from pig brain (PBL, polar brain lipid) is used. This lipid simulates the phospholipid membrane of the brain capillary endothelium. In the MDCK model, the membrane...
|
guest ::
