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Synthesis of unsymmetrical derivatives of azaphthalocyanines and their complex formation with pyridine
Žižkovská, Jana ; Zimčík, Petr (advisor) ; Miletín, Miroslav (referee)
1. Summary Synthesis of unsymmetrical derivatives of azaphthalocyanines and their complex formation with pyridine Mgr. Jana Žižkovská Department of Pharmaceutical Chemistry and Drug Control Faculty of Pharmacy in Hradec Králové, Charles University in Prague I prepared the mixture of six possible products (AAAA, AAAB, ABAB, AABB, ABBB, BBBB) using a statistical condensation starting from two different precursors - 5,6- bis(diethyamino)pyrazine-2,3-dicarbonitrile (A) and 5-diethylamino-6-[2-(4,4'- dimethoxytrifenymethoxy)-ethylamino]pyrazine-2,3-dicarbonitrile (B). Only compound of AAAB type was isolated by column chromatography from the mixture, purified and was characterized by NMR, IR, MS, UV-VIS spectroscopy. The precursor (B) was changed - 5-diethylamino-6-[2-(4,4'-dimethoxytrifenymethoxy)- ethylmethylamino]pyrazine-2,3-dicarbonitrile - and the statistical condensation was done the same way. Unfortunately the desired AAAB product was unstable and therefore the work was finished. In another part of this work I have studied a formation of proton-transfer complex with two molecules of pyridine. Three different AzaPc with hydroxy groups on periphery were used in this study. The influence of hydroxy groups on the rate constant was not confirmed. Then I used only one AzaPc -...
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Laser cooling of the trapped ions for frequency and time metrology
Číp, Ondřej ; Pham, Minh Tuan ; Čížek, Martin ; Lešundák, Adam ; Hucl, Václav ; Hrabina, Jan ; Řeřucha, Šimon ; Jedlička, Petr ; Lazar, Josef
In the laser cooled trapped ions field current research is oriented to yield isolated ions in their basic state of the motion. The detection of the Doppler cooled iont excitation to its quadrupole transition of the electronic structure gives the opportuniny to stabilize highly coherent lasers of the optical frequency of the hundreds of THz. This way a new standart of the time or optical frequencies called „Optical atomic clock“ can be defined. Institute of the Scientific Instruments in Brno in the cooperation with the Department of the Optics of the Palacky University in Olomouc implemented a unique research infrastructure for laser cooling of 40Ca+ ions and subsequent experiments of the quantum mechanics and spectroscopy in a joint laboratory in Brno. Currently 40Ca+ ions are routinely generated and captured. The Doppler cooling of those ions is performed using dipole transition at a wavelength of the 397 nm. Also detection and spectroscopy of the electronic structure of the ion and the presence of his dark resonances is implemented.\n
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Biochemical methods as tool for study of reproductive proteins
Postlerová, Pavla ; Zigo, Michal ; Pohlová, Alžběta ; Jonáková, Věra
Study of molecular mechanisms in reproduction is essential for the understanding of this outstanding process. Our lab studies proteins secreted by reproductive organs and sperm using various biochemical methods for a long time. We have expertise in protein extraction from spermatic cells using different approaches, and by kits for proteins from the sperm surface and distinct subcellular compartments. The proteins of reproductive organ fluids are separated by chromatographic methods, such as size exclusion chromatography, high-performance liquid chromatography with reverse phase (RP-HPLC) and affinity chromatography on matrices with various ligands. Proteins are subjected to SDS- or 2D-electrophoresis for their characterization and comparison of various extraction methods, different mammalian species, and sperm in different functional development. Electrophoretically separated proteins may be transferred onto nitrocellulose membrane (Western blot) for antibody detection or binding studies with lectin-labelled ligands (lectins, polysaccharides, zona pellucida glycoproteins). We use immunoprecipitation method with specific antibody for protein determination followed by the MALDI identification. Proteins are localized by immunofluorescent techniques on/in spermatic cells and tissue sections of reproductive organs. Isolation of proteins from reproductive tissues and fluids, and the antibody detection is crucial for the studying of reproductive protein origin.
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Nursing care in a hygienic and epidemiological regime from the patient's perspective.
KŘEPELOVÁ, Lenka
Considering contemporary lifestyle and possibility of travelling imported infections accrued and some illnesses come back that have not occurred nearly at us more. Not only haemorrhagic fevers and other highly infectious illnesses but also hospital multiresistant bacterial strains that mean high risk for patients constitute a threat. The number of nosocomial infectious grows still. The patient is in the case of suspicion or already diagnosed infectious illness isolated and treated in the infectious department or in the tribal department in the room with increased hygienic-epidemiologic regime. The patient is saved in a single room, the possibility of visits is reduced and the staff uses personal protective equipment. The man is holistic being and it can come due to isolation to lack of satisfaction of his/her bio-psycho-social needs. The aim of this thesis is to identify unsatisfied bio-psycho-social needs of patients in increased hygienic-epidemiologic regime and to describe changes in needs of the patient depending on the time of hospitalisation. The theoretic part describes the problematics of infectious illnesses, system of the care for the patient with infectious illness and changes in human needs in the period of the illness. The empiric part of the thesis deals with unsatisfied bio-psycho-social needs of isolated individual through quantitative research. The research suite was made up of 10 patients who were hospitalised in the room with increased hygienic-epidemiologic regime. The survey was carried out in the Hospital Jihlava by semistructured interview and the results of the survey were processed by opened coding by method paper and pencil. It is emphasized that the most needs of the patient have biological, psychical, social and even spiritual aspects. The rate of satisfaction with caregiving depends on the health condition and personality of the patient. Not small influence has the length of the hospitalisation, too. It was found out during the survey that the patients have not enough information about specifics of the care in the infectious department and that is why there is often a misunderstanding of necessity of barrier measures. It is possible to avoid useless misunderstanding through timely and suitable education. It was created an educational material with the title "Guide for patients hospitalised in the infectious department of Hospital Jihlava" for this purpose. Employees of the infectious department of Hospital Jihlava were informed with results of the research during a workshop, too.
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The study of the role of biotransformational enzymes in chemical carciogenesis
Kondrová, Eliška ; Souček, Pavel (advisor) ; Skálová, Lenka (referee) ; Vobořilová, Jana (referee) ; Bořek Dohalská, Lucie (referee)
In vitro biotransformation studies are an integral part of both toxicological research and drug development. They allow for a significant reduction of tests on human volunteers and provide detailed information about the metabolism of a given compound. Due to limited availability of human liver tissue, it is necessary to make use of alternative model systems and model animal species in the study of the interactions between biotransformation enzymes and xenobiotics. Since the activities of the most important human biotransformation enzymes are absent in the most commonly used laboratory species, rat, information gained in experiments with minipigs is very valuable. Recombinant human biotransformation enzymes expressed in bacteria can be used in biotransformation studies in the form of isolated bacterial membranes. These model systems provide information about the metabolism of a given xenobiotic by a defined enzyme and thein advantages compared to purified enzymes include low cost and quicker and easier preparation with lower loss of enzymatic activity during isolation. The aims of this study were: 1) To prepare a model system with recombinant human biotransformation enzymes expressed in bacterial membranes and to compare the properties of this system with minipig liver microsomes. To assess the usefulness of...
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Stability of vacuoles isolated from Scutellaria baicalensis Georgii suspension cultures
Šmolíková, Cecílie ; Martin, Jan (advisor) ; Tůmová, Lenka (referee)
Cecílie Šmolíková, Stability of vacuoles isolated from Scutellaria baicalensis Georgii suspension cultures, diploma work, Charles University in Prague, Pharmaceutical Faculty in Hradec Králové, trainer of diploma work: Pharm. Dr. Jan Martin Ph.D., Hradec Králové, 2008, 49 pages The ambition of my work was to develop the methodology for purification of vacuoles from Scutellaria baicalensis Georgii suspension cultures and determination of their stability in different solutions. My work is a continuation of the diploma work of Šárka Ševčíková.(59) For my work I used her final solution of vacuoles from which I removed other organelles, the remains after lysis of cellular walls and the plasmatic membranes. The best solution for storage of the purified vacuoles proved to be the solution made up from: 500 mM manitol; 10 mM hepes pH= 7, 4; 5 % ficoll 400. Key words: purification of vacuoles, Scutellaria baicalensis, neutral red, baicalin, baicalein, ficoll 400, tonoplast
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Isolation of vacuoles from Scutellaria baicalensis Georgii suspension cultures
Ševčíková, Šárka ; Martin, Jan (advisor) ; Dušek, Jaroslav (referee)
10. ABSTRACT Šárka Ševčíková Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Heyrovského 1203, 500 05 Hradec Králové Isolation of vacuoles from Scutellaria baicalensis Georgii suspension cultures Diploma thesis Supervisor: Pharm.Dr. Jan Martin, Ph.D. Page Count: 52 The goal of the thesis was to develop methodology for isolation of vacuoles from suspension cultures of Scutellaria baicalensis Georgii of the Lamiaceae family. The original concept was based on a method used at the Division of Pharmacognosy, Leiden University, Holland. This method proved unsuitable. After getting acquainted with available papers concerning methods of isolation of vacuoles from miscellaneous plant species and after performing a series of experiments, we have managed to introduce such method of our own which utilizes Ficoll gradient. Preexisting methods of isolation of vacuoles varied in data on lytic enzymes, incubation time, lytic solution composition and density gradient, too. We managed to optimize the individual steps, which led to successful isolation of vacuoles. The resulting method makes it possible to isolate vacuoles in a relatively short time and in sufficient quality for further experiments. Keywords: Baikal Skullcap, Scutellaria, vacuoles, flavonoids, ficoll, gradient, isolation of vacuoles
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CEA in breeds used for dog-assisted activities and therapies
Tipková, Zuzana ; Sedláková, Vladimíra (advisor) ; Fiala Šebková, Naděžda (referee)
Collie Eye Anomaly (CEA) is an inherited disease that primarily affects herding dog breeds such as Smooth and Rough Collie, Border Collie, Shetland Sheepdog and Australian Shepherd, as well as Nova Scotia Duck Tolling Retriever. All these breeds can be used for dog-assisted activities, some individuals are also suitable for dog-assisted therapies. In the literature I found general information about the history, symptoms, diagnosis, mode of inheritance, genetic testing of the predisposed breeds.
CEA was first described in the 50s of the 20th century. Manifestations of CEA vary widely and therefore it has not been known that a variety of symptoms is caused by one disease for a long time.
Choroidal hypoplasia is the characteristic sign, furthermore may be present: coloboma, tortuosity of the retinal blood vessels, intraocular hemorrhage and retinal detachment leading to blindness. CEA cannot be cured. It is recommended to perform an early diagnosis, which is possible from 6 to 8 weeks of age, because the defect can be covered by later pigmentation in older pups and ocular fundus of the dog then appears normal. At first dog breeders did not pay much attention to this anomaly, because it is a small percentage of individuals who become completely blind as a consequence of that disease. Carriers of CEA have no symptoms, but their offspring may inherit a severe form of CEA. We cannot exclude affected individuals from breeding program in the breeds with high prevalence of CEA since it would reduce genetic diversity.
It is an autosomal recessive disease caused by mutation in the NHEJ1 gene. After the causative mutation was found, a two-step PCR assay, which uses two primer pairs to quickly detect the mutation in chromosomes, was developed. The latest molecular methods for the diagnosis of CEA include simplified analysis of the deletion without DNA isolation and rapid genotyping technique based on SYBR Green real-time PCR. Genetic tests are available in many countries now and so breeders can control the health of their dogs.
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