|
|
Preparation and characterization of nanoliposomal carriers of hydrophobic cytostatics using nanofluidic mixing
Zelníčková, Jaroslava ; Mašek,, Josef (referee) ; Turánek, Jaroslav (advisor)
This diploma thesis is focused on preparation of liposome by relatively new method called nanofluidisation. This method allows the controlled preparation of small unilamellar liposomes in one step. In my thesis I was dealing with optimalization of liposomes preparation which carry hydrophobic cytostatics using this method. Cytotoxic effect of liposomes carrying hydrophobic cytostatics in vitro on cell lines A549 and MCF-7 was determined. In cytotoxicyty test I compared the effect of hydrophobic cytostatics (paclitaxel and derivates of vitamin E specifically alfa-Tos, alfa-TEA) that were incorporated into liposomes prepared via nanofluidisation method and lipid film hydration method. Moreover, a technology of lyophilisation in the presence of cryoprotectants for preparation of liposomes using the method of nanofluidisation was developed.
|
|
|
Structural studies of selected protein complexes involved in signal transduction
Honzejková, Karolína ; Obšil, Tomáš (advisor) ; Bouřa, Evžen (referee) ; Pavlíček, Jiří (referee)
Protein-protein interactions are critical for most physiological and pathophysiological processes. Detailed characterization of these interactions is therefore essential not only to understand the nature of these events, but also to design strategies to target these interactions. This work focuses on the study of the structure and interactions of several proteins and their complexes. Apoptosis signal-regulating kinase 1 (ASK1) is a mitogen-activated protein kinase kinase kinase (MAP3K) that activates the p38/JNK protein kinase pathways, thereby directing cells toward an inflammatory response or apoptosis. ASK1 interacts with thioredoxin (TRX), a small dithiol oxidoreductase, which inhibits ASK1, but the mechanism of this inhibition has not been clarified. CaMKK1 and CaMKK2 are Ca2+ /calmodulin (CaM)-dependent protein kinases that regulate cellular energy balance, memory, and inflammation, among others. Both are inhibited by 14-3-3 proteins, but despite their domain and sequence similarities, the extent of 14-3-3 protein- mediated inhibition is different. Estrogen receptor alpha (ERα) is a nuclear receptor involved in breast cancer. Tamoxifen, an ERα antagonist, is used to treat this disease, but resistance often develops. 14-3-3 proteins interact with ERα and inhibit its transcriptional activity,...
|
|
|
Structural and functional characterization of the E3 ligase BIRC6
GRATZL, Sascha
The aim of the thesis was to characterize the ubiquitin ligase BIRC6 in several contexts. The goal was to expand the knowledge on the selectivity of E1-E2 recognition, to increase the known substrate range of BIRC6, and to elucidate the molecular interactions fulfilled by BIRC6 during autophagy via interaction with GABARAP. In addition, it was aimed to gain a better understanding on the general ubiquitination mechanism in E2/E3 hybrid enzymes, to identify the ubiquitination sites of known apoptotic and autophagic substrates and to enlighten substrate recognition by BIRC6.
|
|
|
Pathofysiology of a tumour microenvironment of salivary glands cancer
Kuchař, Martin ; Skřivan, Jiří (advisor) ; Pokorný, Jaroslav (referee) ; Lohynská, Radka (referee)
Treatment options for salivary gland carcinomas (SGC), especially advanced ones, are limited. Immunotherapy, particularly therapy with immune checkpoint inhibitors (ICI), has brought significant progress and change in the treatment of malignant tumors. The effect and response to immunotherapy using ICI are largely driven by the characteristics of immune cells in the tumor tissue and, as it turns out, also in the peritumoral tissue. We conducted an immunohistochemical analysis of the expression of the immune checkpoint protein PD-1 and its ligand PD-L1 on the surface of tumor cells as well as tumor-infiltrating immune cells (TIIC) in samples of salivary carcinomas, separately in their centre and at their periphery. In addition to the above, an increasing amount of evidence suggests that resistance to ICI therapy is modulated by the interaction of the Fas receptor (CD95) and Fas ligand (FasL, CD178) between tumor cells and immune cells. We therefore decided to explore the expression and interaction of Fas-FasL between tumor cells and tumor-infiltrating immune cells in the centre of the tumor and in the peritumoral area of salivary carcinoma samples. Differential evaluation of the tumor centre and tumor periphery across various histological subtypes of SGC revealed the role of peripheral TIICs and...
|
|
|
Amaryllidaceae alkaloids of haemanthamine structural type and their semisynthetic derivatives as potential drugs in the treatment of Alzheimer's disease.
Peřinová, Rozálie ; Cahlíková, Lucie (advisor) ; Lapčík, Oldřich (referee) ; Mučaji, Pavel (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany Candidate: Ing. Rozálie Peřinová Supervisor: prof. Ing. Lucie Cahlíková, Ph.D. Title of doctoral thesis: Amaryllidaceae alkaloids of haemanthamine structural type and their semisynthetic derivatives as potential drugs in the treatment of Alzheimer's disease. To deepen the knowledge about the Amaryllidaceae alkaloid haemanthamine, which was isolated at our workplace as part of previous phytochemical studies, derivatives of this alkaloid were synthesized. First, series of aliphatic (3-12) and aromatic ester derivatives (13-66) were prepared, and then, to compare the structure-activity relationship, ether derivatives (67-80) were prepared from the most active substituents. All synthesized compounds were identified using the following structural analysis methods: NMR, HPLC/MS, and HRMS, including testing physical properties such as optical rotatability. After structure confirmation, all derivatives were subjected to screening studies for their inhibitory potential against hAChE and hBuChE. The selected derivatives were tested for their inhibitory potential against another enzyme, GSK-3β, which plays a significant role in the pathogenesis of AD. In cooperation with the Faculty of Medicine in...
|
|
|
The effect of synthetic modified mRNAs induced proliferation on pancreatic beta cells
Veľasová, Adriana ; Koblas, Tomáš (advisor) ; Bořek Dohalská, Lucie (referee)
Diabetes mellitus is a chronic disease caused by the loss of pancreatic beta cells due to autoimmune destruction or increased apoptosis. Beta-cell deficiency results in reduced insulin production, which plays an important role in glucose metabolism. The number of beta-cells in the body is one of the main factors that influence the development of this chronic disease. Therefore, it is necessary to find a way by which the number of beta-cells of the organism can be increased and thus the insulin production can be restored in a natural way without any need for the use of insulin infusions. However, the ability of beta-cells to divide decreases with age and is virtually nil in adulthood. The study of the cell cycle, especially the early and late cyclins and cyclin-dependent kinases, which act as cell cycle regulators, thus appears to be a promising way to restore natural insulin-producing tissues. In order to increase the number of beta cells entering the cell cycle, we focused on studying the effect of in vitro transcribed (IVT) mRNAs, encoding cyclins type D and cyclin dependent kinases 4 and 6 on stimulating cell division of isolated beta-cells. We found that transfection IVT mRNAs for type D cyclins in combination with cyclin-dependent kinases 4 and 6 significantly increased the proliferation of beta-cells...
|
|
|
The role of a specific miRNAs in the regulation of apoptosis during physiological and pathophysiological processes in the CNS
Kaslová, Tereza ; Romanyuk, Natalyia (advisor) ; Klassen, Ruslan (referee)
MicroRNAs are small non-coding RNAs of 20 to 24 nucleotides in size that are able to post- transcriptionally regulate gene expression by binding to mRNA. This paper focuses on how these microRNAs are generated and how they are able to regulate at the level of proteins involved in programmed cell death - apoptosis. By what mechanisms apoptosis occurs, what proteins are involved and what changes the cell undergoes are further discussed in this thesis. The precise influence of this post-transcriptional regulation is presented by using selected microRNAs that influence apoptosis during the development of the central nervous system, as well as during and as a consequence of the neurodegenerative diseases and damage that can affect it. Finally, it will also introduce the use of microRNAs as potential biomarkers, due to changes in their levels associated with various diseases, and as direct therapeutic targets. Keywords Apoptosis, microRNA, cell death, central nervous system, neurodegenerative diseases, gene expression regulation
|
|
|
PREPARATION AND CHARACTERIZATION OF NATURAL EXTRACTS WITH ANTIMICROBIAL EFFECT
Vysoká, Marie ; Obruča, Stanislav (referee) ; Kráčmar, Stanislav (referee) ; Márová, Ivana (advisor)
Celosvětová spotřeba antibiotik v posledních několika desetiletích významně vzrostla, což vedlo ke zvýšenému výskytu genů antibiotické rezistence (ARG), které představují rostoucí vážnou hrozbu pro lidské zdraví na celém světě. Používání přírodních nebo alternativních léčivých přípravků tudíž v posledních několika letech výrazně vzrostlo. Velká pozornost je nyní věnována přírodním extraktům, které by mohly být v jisté formě použity jako jedna z možností léčby bakteriálního onemocnění, a to s vysokou účinností proti patogenům a se zanedbatelnými vedlejšími účinky. Biomasa kvasinek, která obsahuje cenné mastné kyseliny, sacharidy, nukleové kyseliny, vitamíny a minerály, byla testována jako možná alternativa, a to samotná i v kombinaci s rostlinnými extrakty ve formě doplňku stravy. Tato disertační práce je zaměřena na studium antimikrobiální aktivity a nutriční přínosy vybraných rostlinných extraktů, jako je acai, hřebíček, kurkuma, zázvor, ženšen a oregano. Následně byly testovány biologické účinky kvasinkové biomasy rodu Metschnikowia pulcherrima CCY 029-002-145, Cystofilobasidium infirmominiatum CCY 17-18-4, Phaffia rhodozyma CCY 77-1, Rhodotorula kratochvilovae CCY 20-2-26. Tyto kvasinky byly vybrány zejména pro vysoký obsah aktivních látek v produkované biomase. Na závěr byl posouzen potenciální synergický efekt rostlinných extraktů ve spojení s kvasinkovou biomasou ke zvýšení celkového antimikrobiálního účinku, antioxidační aktivity a vlivu na růst buněčných linií Caco-2, B16F1 a HaCaT. Vzhledem k dosaženým výsledkům lze konstatovat, že zařazení imortalizované kvasinkové biomasy v kombinaci s přírodními extrakty do doplňků stravy ve vhodné formě může být prospěšné lidskému zdraví. Existuje přímý vztah mezi životaschopností buněk a apoptózou. Všechny mikrobiální extrakty vykazovaly určité pozitivní výsledky na apoptózu u rakovinných buněčných linií a současně nevykazovaly žádné cytotoxické účinky na nezhoubné buněčné linie při stejných koncentracích.
|
|
|
Molecular mechanisms of cfDNA effects
Hříbková, Natálie ; Daňková, Pavlína (advisor) ; Škubica, Patrik (referee)
The biological role of free circulating DNA is currently being investigated mainly in terms of the etiology of autoimmune conditions and cancer. It appears that the role of cfDNA of nuclear or mitochondrial origin in interacting with receptors of the innate immune system is quite complex and cfDNA is thus involved in a variety of molecular mechanisms such as the spread of cancer metastasis, induction of autophagy and triggering of inflammatory processes. Knowledge of the specific pathways in which cfDNA is involved in the development of these pathologies seems to be crucial, especially in terms of selecting and targeting appropriate therapies. This paper provides a framework for the knowledge on the interaction of cfDNA with receptors of the innate immune system, the induction of immunological signaling pathways and the possible consequences that these molecular pathways may have in the body. Key words: cfDNA, TLR9, cGAS/STING, AIM2, autoimmune disease, biomarker, cancer
|
|
|
Mitochondrial dysfunction in brain tumors
Rollerová, Kateřina ; Vaňátko, Ondřej (advisor) ; Zobalová, Renata (referee)
Brain tumors are one of the most serious pathologies of the central nervous system. Brain tumors are aggressive and very hard to treat due to the fragile nature of the nervous system, presence of blood-brain barrier and high recurrence rate. One of the hallmarks of brain tumors is mitochondrial dysfunction. Mitochondria are organelles involved in essential cellular processes, such as energy production, redox and calcium signaling, or the regulation of cell death. Structural and functional abnormalities, mutations in the mitochondrial genome and other mitochondrial dysregulations may cause disruptions in various cellular processes, such as production of reactive oxygen species, migration, proliferation, or regulation of cell death, promoting the development and/or maintenance of brain tumors. The goal of this thesis is to summarize current knowledge about mitochondrial dysfunction in brain tumors. Key words: brain tumors; mitochondria; mitochondrial dysfunction; Warburg effect; apoptosis; reactive oxygen species; isocitrate dehydrogenase
|
guest ::
