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Fylogeneze vybraných druhů letounů Afriky na základě cytogenetického a molekulárního přístupu
Koubínová, Darina ; Zima, Jan (advisor) ; Macholán, Miloš (referee) ; Martínková, Natália (referee)
Phylogenetic relationships of a sample comprising 248 bats belonging to 19 species and four families (Hipposideridae, Rhinolophidae, Molossidae and Vespertilionidae) from Senegal (Western Africa) were investigated with the use of multi-locus sequence data and non- differentially stained chromosomes. The karyotypes of Hipposideros ruber, H. tephrus, H. jonesi and H. cyclops were described for the first time. The standard Hipposideros formula was recorded in H. tephrus, H. jonesi and H. ruber (2n = 32, FNa = 60, FN = 64). The karyotypes of H. cyclops (2n = 36, FN = 66) and H. gigas (2n = 52, FN = 64) substantially diverged from this typical chromosomal complement. Rhinolophus landeri and R. fumigatus shared the same diploid number (2n = 58), but differed in the chromosome morphology (R. fumigatus - FNa = 60, FN = 64; R. landeri - FNa = 64, FN = 68). Rhinolophus landeri was found karyotypically distinct to other African populations, thus signalling a possible presence of cryptic forms within this species. The karyotypes of Chaerephon pumilus and Mops condylurus had a 2n = 48, FN = 54 and were similar to other previously studied species of this chromosomally conservative family. Chromosomal, Bayesian, maximum likelihood and genetic distance analyses revealed an indication for the existence of cryptic...
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Development of a bioanalytical method for the determination of novel drug candidates derived from thiosemicarbazone in a biological material
Suprunová, Vlasta ; Stariat, Ján (advisor) ; Kučera, Radim (referee)
Nowadays, high-performance liquid chromatography is widely used analytical method for separation and quantification of drugs in biological material due to its high sensitivity and selectivity. HPLC is suitable for quantitative as well as qualitative analysis. 2-benzoylpyridine-4-ethyl-3-thiosemicarbazone (Bp4eT) is a potential drug from the group of thiosemicarbazones, which are currently intensively studied and developed as anticancer agents. Their specific mechanism of action, based on chelation of iron, might overcome resistance to standard chemotherapy. The aim of this study was to develop and assess essential validation parameters of an HPLC-MS/MS method for determination of Bp4eT and its main phase I metabolites in rat plasma and to utilise the method for the analysis of samples from a pilot pharmacokinetic study in rats. The separation of Bp4eT and its metabolites was achieved on chromatographic column Discovery HSC18 (75 x 4.6 mm, 3 µm) protected by the same type of guard column using the mobile phase consisting of ammonium formate and acetonitrile in ratio 40:60 (v/v). The isocratic elution and the flow rate of 0.4 ml/min were utilised. Mass spectrometer was chosen as a detector and the quantification was performed in selected reaction monitoring mode. Plasma samples were treated using the...
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New matrices for MALDI-MS analysis of lipids
Macháčková, Petra ; Cvačka, Josef (advisor) ; Volný, Michael (referee)
In this work focused on research of new matrices for MALDI-MS analysis of lipids several organic salts were tested as potential matrices for hydrocarbons and wax esters. It was shown previously that the effective ionization of lipids occurs in conjuction with a lithium cation to form adducts [M + Li] + . Therefore, lithium salts of aromatic acids were synthesized. Matrices were evaluated according to their MALDI properties and compared with lithium 2,5-dihydroxybenzoate, which is proven matrix for MALDI analysis of lipids. It was found that effectiveness of matrix depends on presence of hydroxyl group in matrix molecule. The best results were achieved with lithium vannilate, even in comparison with lithium 2,5-dihydroxybenzoate.
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Molecular phylogeny and evolutionary trends in Hieracium (Asteraceae, Lactuceae)
Krak, Karol ; Fehrer, Judith (advisor) ; Zozomová, Judita (referee) ; Nieto Feliner, Gonzalo (referee)
The hawkweed subgenus Hieracium s. str. is notoriously known for its extreme morphological variability and variation in ploidy levels that is associated with differences in modes of reproduction. Extensive past hybridization is supposed for the subgenus, but recent hybridization was evidenced only in few cases. The subgenus attracts the attention of botanists already for more than a century. Therefore the species diversity is largely examined and the taxonomy of the subgenus is well elaborated, although several contradictory taxonomic concepts exist. However the relationships among the species are unknown and haven't been studied yet. The investigation of these relationships from a phylogenetic perspective using molecular approaches was the main aim of the presented thesis. Basic species (both diploid and polyploid), representing morphologically unique taxa, that are supposed to be the basic evolutionary units of the subgenus were studied. The sequences of two intergenic spacers of the cpDNA (trnT-trnL and trnV-ndhC) and the external transcribed spacer of the nuclear ribosomal DNA (nrDNA ETS) were analyzed. Moreover, three new low-copy nuclear markers with higher variability than nrDNA and cpDNA markers were developed and their suitability for phylogenetic studies in Hieracium s. str. was...
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Analysis of antimicrobial peptides in venom glands of bumblebees.
Janechová, Daniela ; Cvačka, Josef (advisor) ; Monincová, Lenka (referee)
The growing resistance of bacteria to traditional antibiotics promotes the interest in finding new substances for their production. Antimicrobial peptides have comparable effect to conventional antibiotics, but a different mechanism of action and they do not provoke bacterial resistance. These peptides were characterized in all forms of multicellular organisms. Hymenoptera venom contains many biologically active substances including antimicrobial peptides. For this reason, this thesis focuses on the acquisition of antimicrobial peptide sequences from selected species of bumblebees (Bombus terrestris, B. hortorum, B. hypnorum, B. pratorum, B. lucorum, B. lapidarius, B. humilis and B. bohemicus). The isolation from the venom glands was performed by high performance liquid chromatography with reversed phases. Subsequent analysis was performed using the methods of mass spectrometry, matrix-assisted laser desorption/ionization with time of flight analyzer and electrospray ionization connected with hybrid linear ion trap analyzer with orbitrap. The sequences for the found peptides were determined by tandem mass spectrometry methods "de novo" and Edman degradation. In this work we characterized 17 sequences of peptides extracted from bumblebee venom glands for which antimicrobial activity was determined...
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Lifestyle and relationships in the subculture of ballet
Erben, Jakub ; Buriánek, Jiří (advisor) ; Sládek, Jan (referee)
This thesis deals with the lifestyle and relationships among professional ballet dancers. Previous studies that have focused on this group, have not focused on the lifestyle as a complex phenomenon, the goal is therefore to supplement existing knowledge in this matter. Given the recent research coverage problem, the thesis uses explorative approach to the conceptual level based on the concepts of lifestyle and subculture and it uses methodologically a qualitative approach. In particular, the conceptual basis in the form of subculture largely determines the orientation, the process and the structure of the paper. It figures as a tool that allows us to organize a view of professional ballet dancers. After defining the concepts related to ballet and introducing the factual account of events enabling to look into the historical and institutional context in which Czech ballet dancers act, followed by a description logic of its study in this paper, including basic concepts of introducing subculture and lifestyle, and clarification of the used methodology. The results of qualitative research among professional ballet dancers that form the core of the paper, are then presented in the next section. Among ballet dancers there were then monitored various aspects of their lives and lifestyles. As an important...
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Theoretical study of spin-orbit coupling on spectra and photophysics of rhenium complexes
Heydová, Radka ; Záliš, Stanislav (advisor) ; Slavíček, Petr (referee) ; Srnec, Martin (referee)
Title: Theoretical study of spin-orbit coupling on spectra and photophysics of rhenium complexes Author: RNDr. Radka Heydová Department: Physical and Macromolecular Chemistry Supervisor: Ing. Stanislav Záliš, CSc., JHI AS CR, v.v.i. Supervisor's e-mail address: stanislav.zalis@jh.inst-cas.cz Abstract: Relativistic effects, especially spin-orbit coupling (SOC), play an essential role in transition metal chemistry and SOC treatment is indispensable for a correct theoretical description. To demonstrate the importance of SOC, the energies and oscillator strengths of vertical transitions for a series of [ReX(CO)3(2,2'-bipyridine)] (X = Cl, Br, I) and [Re(imidazole)(CO)3(1,10-phenanthroline)]+ complexes were calculated in the spin-free (SF) and spin-orbit (SO) conceptual frameworks. Two different computational approaches were adopted: SO-MS-CASPT2 where SOC was added a posteriori using a configuration interaction model (SO-RASSI), and the approximate perturbative SO-TD-DFT method. Relativistic effects were included via the two-component Douglas-Kroll-Hess transformation and the zeroth-order regular approximation in the former and the latter technique, respectively. The SF (i.e. accounting only for the scalar relativistic effects) and SO results from both methods were compared with each other and to available...
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Education and development of children in lesbian family
Sedláčková, Anna ; Šulová, Lenka (referee) ; Gillernová, Ilona (referee)
The thesis deals with topic of lesbian families and specifics of children from these families. The theoretical part of the work is supported by studies from the USA and the Great Britain in particular. This part is focused on definition of homoparental family and its types, specifics of upbringing and relations within. It also deals with the topic of coming-out. It gives account of the family roles - biological mother, biological father, social mother / stepmother. The second half of the theoretical part describes psychological, gender and social development of the children from lesbian families. There are also mentioned some children's feelings and views of their nontraditional family and homosexuality of their parents. The thesis also deals with lesbian adoptive families. The research presented in the empirical part is based on individual interviews and projective techniques. The study group comprises 10 children between the age 4 and 15. In the study group there are children from previous heterosexual relationship as well as children from planned lesbian families. Participants to the sample were recruited via advertising and snow ball technique. The research is qualitative and its outcomes are elaborated in the form of case study report. The list of crucial themes elaborated through clustering...
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Studies of the Mouse polyomavirus: properties of the minor structural proteins, requirements for virion productive trafficking to the nucleus and observed side effects of DNA transfection.
Huérfano- Meneses, Sandra ; Forstová, Jitka (advisor) ; Mělková, Zora (referee) ; Reiniš, Milan (referee)
In this thesis, we aimed to investigate the role of Mouse polyomavirus (MPyV) minor structural proteins, VP2 and VP3 in the viral life cycle as well as to study specific aspects of the productive trafficking of the virus. The thesis is divided into 3 chapters as follows: chapter 1 addresses questions about the interaction of the minor structural proteins, VP2 and VP3, of MPyV with host cells and their role during late stages of virus infection. In the chapter 2, aspects concerning the role of late endosomes, recycling endosomes and actin cytoskeleton in MPyV productive infection are addressed and in the chapter 3, results of studies of interferon (IFN) responses induced after DNA nucleofection are presented. The study in chapter 3 was preceded by a microarray analysis primary performed to reveal cell responses to the presence of the minor proteins within host cells. We showed that both MPyV minor structural proteins, VP2 and VP3, when produced individually (without VP1) are highly cytotoxic, inducing fast, caspase dependent apoptosis. Immuno-electron and confocal microscopy revealed affinity of both proteins to endocellular membranes. Both VP2 and VP3 were found to be bound to damaged ER and mitochondrial membranes. Interaction of the proteins with membranes causing physical damage of organelles...
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The preparation of tablets by selective laser sintering.
Ficek, Lukáš ; Mužíková, Jitka (advisor) ; Šnejdrová, Eva (referee)
Charles University, Faculty of pharmacy in Hradec Králové Department of: Pharmaceutical Technology Mentor: doc. PharmDr Jitka Mužíková, Ph. D Consultant: Mgr. Thao Tranová Student: Lukáš Ficek Title of Thesis: The preparation of tablets by laser sintering This work focuses on the preparation of orally dispersible tablets by selective laser sintering (SLS). The present project is a pilot study conducted on the 3D printer Sintratec Kit at the Department of Pharmaceutical Technology. SLS is a 3D printing method based on the fusion of powder particles using a laser beam that sinters the individual particles together in the layers of material.1-2 The matrix forming polymer used in work was Kollidon® VA64 which was combined with two pigments namely Candurin® Gold Sheen and NTX Ruby Red. In addition, mixtures of the polymer with a co-processed dry binder Prosolv® ODT G2 and a physical mixture of mannitol, silicified microcrystalline cellulose, and crospovidone at the concentrations of 20%, 40%, and 60% were used. The influence of printing parameters, especially laser speed, on tablet quality was tested. NTX Ruby Red pigment was selected for final tablet printing. The best powder formulation for optimal printing progress and high-quality properties of the orally dispersible tablets was the mixture of...
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