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The Effect of Selected Drugs on Bone Metabolism
Karešová, Iva ; Palička, Vladimír (advisor) ; Bayer, Milan (referee) ; Horák, Pavel (referee)
With an aging population and increasing life expectancy, osteoporosis represents a major global health problem. Currently it is estimated that more than 200 million people suffer from osteoporosis. Fractures due to osteoporosis are the major cause of morbidity and mortality in the elderly (Compston et al., 2019). A large body of evidence suggests an association between AEDs and bone abnormalities. The prevalence of osteoporosis and osteopenia in long-term treated epileptics is high. More than 50% of patients with epilepsy have low bone mineral density (BMD) (Ko et al., 2020). The aim of the dissertation was to evaluate the effect of orchidectomy and the effect of selected new antiepileptic drugs (zonisamide, gabapentin, pregabalin, levetiracetam) on bone metabolism in young Wistar rats. The negative effect on bone tissue after rat orchidectomy was confirmed after 12 weeks. Orchidectomy led to a statistically significant reduction in BMD and had a negative effect on the biomechanical properties of bones. Our results confirm that young rats after orchidectomy can be considered a suitable animal model for the study of osteopenia. After 12 weeks of administration of new antiepileptic drugs (zonisamide, gabapentin, pregabalin, levetiracetam), there was no statistically significant effect on BMD in...
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Epigenetic Regulation of Adhesive Molecules in High-grade Serous Ovarian Carcinoma
Baranová, Ivana ; Palička, Vladimír (advisor) ; Matoušková, Petra (referee)
5 Summary Title: Epigenetic Regulation of Adhesive Molecules in High-grade Serous Ovarian Carcinoma The lack of effective biomarkers for screening and early detection of ovarian cancer is currently considered as one of the most pressing problems in oncogynecology. Because epigenetic alterations occur early in the cancer development, they provide great potential to serve as such biomarkers. Epigenetic mechanisms have been implicated also in regulation of adhesion molecules that play a major role in cancer progression. The main aim of this study was to investigate the methylation pattern of selected cadherin and catenin genes in ovarian cancer tissue by comparison with control tissue. The study group consisted of 68 patients with high-grade serous ovarian cancer (HGSOC) and 46 control patients. To determine the sites with the most significant methylation in selected genes next-generation sequencing was employed. For further confirmation of detected methylation of selected regions, methylation-sensitive high-resolution melting analysis and real-time methylation-specific polymerase chain reaction were used. In attempt to design potential biomarker panel for future screening of HGSOC as the secondary aim of our study, cadherins were evaluated together with transcription factors from our previous study....
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Bioanalytical Evaluation of New Acetylcholinesterase Inhibitors
Mžik, Martin ; Palička, Vladimír (advisor) ; Maier, Vítězslav (referee) ; Kučera, Radim (referee)
6 SUMMARY Bioanalytical evaluation plays a pivotal role in preclinical research of new drugs, particularly in investigation of their pharmacokinetics and metabolism. Use of modern analytical approaches is indispensable; chromatography and mass spectrometry are the techniques of choice in this area of research. To obtain reliable data, the analytical methods must be validated. Several authorities - such as European Medicines Agency or Food and Drug Administration - have published recommendations on bioanalytical method validation. In its theoretical part, this dissertation deals with description of Alzheimer's dementia with special focus on treatment strategies. Further attention is paid to topics that are directly related to the experimental work - liquid chromatography, mass spectrometry, sample preparation, method validation and pharmacokinetics. The experimental part - a commentary on the author's published articles - consists of three units: I. bioanalytical evaluation of PC-37 and PC-48 - novel 7-methoxytacrine and donepezil-derived dual cholinesterase inhibitors, II. study of the transport of donepezil across the blood-brain barrier with focus on its structure and substructures and III. bioanalytical evaluation of S-K1025 and S-K1026 - new 7-methoxytacrine and tryptophan-derived dual cholinesterase...
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Bioanalytical Evaluation of New Acetylcholinesterase Inhibitors
Mžik, Martin ; Palička, Vladimír (advisor) ; Kučera, Radim (referee)
6 SUMMARY Bioanalytical evaluation plays a pivotal role in preclinical research of new drugs, particularly in investigation of their pharmacokinetics and metabolism. Use of modern analytical approaches is indispensable; chromatography and mass spectrometry are the techniques of choice in this area of research. To obtain reliable data, the analytical methods must be validated. Several authorities - such as European Medicines Agency or Food and Drug Administration - have published recommendations on bioanalytical method validation. In its theoretical part, this dissertation deals with description of Alzheimer's dementia with special focus on treatment strategies. Further attention is paid to topics that are directly related to the experimental work - liquid chromatography, mass spectrometry, sample preparation, method validation and pharmacokinetics. The experimental part - a commentary on the author's published articles - consists of three units: I. bioanalytical evaluation of PC-37 and PC-48 - novel 7-methoxytacrine and donepezil-derived dual cholinesterase inhibitors, II. study of the transport of donepezil across the blood-brain barrier with focus on its structure and substructures and III. bioanalytical evaluation of S-K1025 and S-K1026 - new 7-methoxytacrine and tryptophan-derived dual cholinesterase...
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Epigenetic Regulation of Adhesive Molecules in High-grade Serous Ovarian Carcinoma
Baranová, Ivana ; Palička, Vladimír (advisor) ; Matoušková, Petra (referee)
5 Summary Title: Epigenetic Regulation of Adhesive Molecules in High-grade Serous Ovarian Carcinoma The lack of effective biomarkers for screening and early detection of ovarian cancer is currently considered as one of the most pressing problems in oncogynecology. Because epigenetic alterations occur early in the cancer development, they provide great potential to serve as such biomarkers. Epigenetic mechanisms have been implicated also in regulation of adhesion molecules that play a major role in cancer progression. The main aim of this study was to investigate the methylation pattern of selected cadherin and catenin genes in ovarian cancer tissue by comparison with control tissue. The study group consisted of 68 patients with high-grade serous ovarian cancer (HGSOC) and 46 control patients. To determine the sites with the most significant methylation in selected genes next-generation sequencing was employed. For further confirmation of detected methylation of selected regions, methylation-sensitive high-resolution melting analysis and real-time methylation-specific polymerase chain reaction were used. In attempt to design potential biomarker panel for future screening of HGSOC as the secondary aim of our study, cadherins were evaluated together with transcription factors from our previous study....
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Bioanalytical Evaluation of New Acetylcholinesterase Inhibitors
Mžik, Martin ; Palička, Vladimír (advisor) ; Maier, Vítězslav (referee) ; Kučera, Radim (referee)
6 SUMMARY Bioanalytical evaluation plays a pivotal role in preclinical research of new drugs, particularly in investigation of their pharmacokinetics and metabolism. Use of modern analytical approaches is indispensable; chromatography and mass spectrometry are the techniques of choice in this area of research. To obtain reliable data, the analytical methods must be validated. Several authorities - such as European Medicines Agency or Food and Drug Administration - have published recommendations on bioanalytical method validation. In its theoretical part, this dissertation deals with description of Alzheimer's dementia with special focus on treatment strategies. Further attention is paid to topics that are directly related to the experimental work - liquid chromatography, mass spectrometry, sample preparation, method validation and pharmacokinetics. The experimental part - a commentary on the author's published articles - consists of three units: I. bioanalytical evaluation of PC-37 and PC-48 - novel 7-methoxytacrine and donepezil-derived dual cholinesterase inhibitors, II. study of the transport of donepezil across the blood-brain barrier with focus on its structure and substructures and III. bioanalytical evaluation of S-K1025 and S-K1026 - new 7-methoxytacrine and tryptophan-derived dual cholinesterase...
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Epigenetic Regulation of Adhesive Molecules in High-grade Serous Ovarian Carcinoma
Baranová, Ivana ; Palička, Vladimír (advisor) ; Matoušková, Petra (referee) ; Friedecký, David (referee)
5 Summary Title: Epigenetic Regulation of Adhesive Molecules in High-grade Serous Ovarian Carcinoma The lack of effective biomarkers for screening and early detection of ovarian cancer is currently considered as one of the most pressing problems in oncogynecology. Because epigenetic alterations occur early in the cancer development, they provide great potential to serve as such biomarkers. Epigenetic mechanisms have been implicated also in regulation of adhesion molecules that play a major role in cancer progression. The main aim of this study was to investigate the methylation pattern of selected cadherin and catenin genes in ovarian cancer tissue by comparison with control tissue. The study group consisted of 68 patients with high-grade serous ovarian cancer (HGSOC) and 46 control patients. To determine the sites with the most significant methylation in selected genes next-generation sequencing was employed. For further confirmation of detected methylation of selected regions, methylation-sensitive high-resolution melting analysis and real-time methylation-specific polymerase chain reaction were used. In attempt to design potential biomarker panel for future screening of HGSOC as the secondary aim of our study, cadherins were evaluated together with transcription factors from our previous study....
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Epigenetic Alternations in Head and Neck Squamous Cell Carcinoma: Potential Biomarkers
Kovaříková, Helena ; Palička, Vladimír (advisor) ; Friedecký, David (referee) ; Pešta, Martin (referee)
5 1 Summary ENG Head and neck squamous cell carcinomas are a group of heterogenic tumors arising from epithelial tissue of aerodigestive tract characterized by difficult diagnosis, treatment and prognosis. Alcohol consumption, smoking and high-risk human papillomavirus infection are very well described risk factors of head and neck cancer development. Sinonasal carcinomas are group of malignancies developing in nasal and paranasal sinuses. Oropharyngeal carcinomas are malignancies developing in the throat area downstream to oral cavity. microRNAs (miRNAs) are short (~23 nucleotides) non-coding RNA molecules participating in regulation of gene expression. Primary function of miRNAs is negative translation regulation as part of RISC (RNA-induced silencing complex) by translational repression and mRNA degradation. miRNAs are involved in various disease pathologies such as neurodegenerative diseases, metabolic disorders and cancer. Moreover, microRNAs have been recognized as key molecules in cancer development and progression in various types of tumors. My doctoral study research activities were focused on investigation of relative expression of several preselected miRNAs (using real-time PCR) and the relationship between their expression and clinicopathological characteristics of the squamous cell carcinoma...
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Laboratory diagnostics of kidney function
Šálek, Tomáš ; Palička, Vladimír (advisor) ; Jabor, Antonín (referee) ; Průša, Richard (referee)
Laboratory diagnostics of kidney function The work deal with issue of glomerular filtration rate (GFR). GFR is the most important indicator of kidney function. Its decline is associated with increased morbidity and mortality. Chronic kidney disease (CKD) is defined as decrease of GFR bellow 1.0 ml/s/1,73m2 for more than three months. Patients with CKD receive nephroprotective drugs and have reduced drugs excreted by kidneys if needed. Early detection of CKD is of clinical importance. We compare GFR estimated from serum creatinine (eGFRcreatinine) and from serum cystatin C (eGFRcystatinC) in four cohorts of patients: 101 MGUS patients, 13 methanol intoxicated patients, 1515 diabetics and 352 patients with CKD. Creatinine was measured by enzymatic method traceable to international reference material NST SRM 967 in all patients. Cystatin C was determined by standardized immunoturbidimetric method traceable to DA ERM 471. This standardized method was not available only for MGUS patients. eGFR was always calculated according to the best available validated equation. MGUS patients were calculated according to MDRD equation for eGFRcreatinine and according to Grubb equation for eGFRcystatinC. Methanol intoxicated patients were estimated according to Lund-MalmÖ equation for eGFRcreatinine and according to...
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DNA methylation changes in ovarian cancer patients
Chmelařová, Marcela ; Palička, Vladimír (advisor) ; Vávrová, Jiřina (referee) ; Bouda, Jiří (referee)
DNA methylation changes in ovarian cancer patients. In our work we studied DNA methylation changes in ovarian cancer. Due to the aggressive nature of this disease, it is very important to obtain detailed mapping of molecular parameters including epigenetics. These parameters could potentially be used in the diagnostics and treatment of ovarian cancer. To begin with, we focused on establishing and optimizing techniques which are suitable for methylation analysis of DNA isolated from paraffin blocks. The next step was to analyze selected CpG loci primarily of tumor suppressor genes. Formalin-fixed, paraffin-embedded tissue samples from both ovarian adenocarcinomas and normal ovarian tissue were obtained from 109 women (69 patients with ovarian cancer, 40 patients with normal ovarium) treated at the Department of Obstetrics and Gynecology, University Hospital Hradec Kralove. The samples of normal ovary were obtained from patients surgically treated for non-malignant diagnosis. DNA was extracted using a Qiagen DNA extraction kit. We established and optimized two techniques - MSP (Methylation specific PCR) and MS-MLPA (Methylation Specific Multiplex Ligation-dependent Probe Amplification). We discovered that for methylation analysis of poor quality DNA isolated from paraffin blocks MS-MLPA is more...
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