Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Pelnářová Supervisor: Prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Václav Tvrdý, MSc.; Marcel Hrubša, MSc. Title of diploma thesis: Searching of effective cobalt chelators - 8-hydroxyquinolines Cobalt is one of the essential trace elements present in the human body. It forms a part of the organometallic complex of vitamin B12, which is essential for many physiological functions. Both overload and lack of cobalt in the body is associated with pathological conditions. Manifestations of deficiency can lead up to pernicious anemia or hypofunction of the thyroid gland. Cobalt intoxication can occur in an industrial environment, such as by cobalt metal dust during heavy metal processing, or by its release due to corrosion from orthopedic prostheses. Systemic toxicity is manifested by a number of endocrine, cardiovascular and neurological symptoms. The main aim of this diploma thesis was to find effective chelators of cobalt from the group of substances derived from 8-hydroxyquinoline. In vitro spectrophotometric measurement was used to determine the degree of chelation. The effect of chelation was also monitored ex vivo in rat erythrocytes. All tested chelators - 8-hydroxyquinoline,...

v angličtině Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Michela Morkusová Supervisor: Přemysl Mladěnka, Pharm.D., Ph.D. Title of diploma thesis: Chelation ability of experimentally used iron chelators Iron is a vital element, playing many important functions in the body. Its presence in too low or too high quantities is harmful. The body has no excretion route for iron, and in certain circumstances, excess iron can accumulate in body tissues. Iron is present in the body in excess particularly in individuals undergoing frequent blood transfusions or having genetically based diseases associated with iron overload (hemochromatosis). Iron excess is medically treated by administration of iron chelators. Such substances can find wider therapeutic use as well, e.g. in individuals with acute myocardial infarction or with tumours. This Thesis was aimed at determining the stoichiometric ratio in the complexes of two experimental chelators, salicylaldehyde isonicotinoyl hydrazone (SIH) and ethylenediaminetetraacetic acid (EDTA), with iron(II) and iron(III) ions at four different pathophysiologically significant pH levels (4.5, 5.5, 6.8 and 7.5). Spectrophotometry was used as the analytical method. In the processing stage, various new...

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Lucie Kovářová Supervisor: Ing. Vladimír Kubíček, CSc. Title of Diploma Thesis: Multicomponent analysis This diploma thesis deals with the determination of concentrations of individual dyes in mixtures by multicomponent spectrophotometric analysis. The method was tested on selected dyes which were divided into two groups. The first group contains methyl orange, methyl red, orange II and phenol red. The second group consists of methylene blue, bromocresol green and thymol blue. Within each group mixtures were prepared containing combinations of two, three and four dyes in various ratio. Spectra of prepared mixtures were measured by diode array UV/VIS spectrophotometer. Absorbance values were read at wavelengths of absorption maxima of the individual dyes. Using the pseudoinverse matrix, concentration ratios of components were calculated from the absorbance values. The determination in determined and over determined systems was performed. On the basis of the results obtained by the method of analysis it is possible to determine the individual concentration of the multi-component dyes mixtures without prior separation. Keywords: spectrophotometry, multicomponent analysis,...

Populärwissenschaftlicher Abstract: <br /> <br /> Bislang gibt es in der beobachtenden optischen Astronomie zwei verschiedene Herangehensweisen: Einerseits werden Objekte durch Kameras abbildend erfaßt, andererseits werden durch die wellenlängenabhängige Zerlegung ihres Lichtes Spektren gewonnen. Das Integral - Field - Verfahren ist eine relativ neue Technik, welche die genannten Beobachtungsmethoden vereint. Das Objektbild im Teleskopfokus wird in räumlich zerlegt und jedes Ortselement einem gemeinsamen Spektrografen zugeführt. Hierdurch wird das Objekt nicht nur zweidimensional räumlich erfaßt, sondern zusätzlich die spektrale Kompenente als dritte Dimension erhalten, weswegen das Verfahren auch als 3D-Methode bezeichnet wird. Anschaulich kann man sich das Datenresultat als eine Abbildung vorstellen, in der jeder einzelne Bildpunkt nicht mehr nur einen Intensitätswert enthält, sondern gleich ein ganzes Spektrum. Diese Technik ermöglicht es, ausgedehnte Objekte im Unterschied zu gängigen Spaltspektrografen komplett zu erfassen. Die besondere Stärke der Methode ist die Möglichkeit, die Hintergrundkontamination der unmittelbaren Umgebung des Objektes zu erfassen und in der Auswertung zu berücksichtigen. Durch diese Fähigkeit erscheint die 3D-Methode prädestiniert für den durch moderne Großteleskope erschlossenen Bereich der extragalaktischen Stellarastronomie. Die detaillierte Untersuchung aufgelöster stellare Populationen in nahegelegenen Galaxien ist erst seit kurzer Zeit dank der Fortschritte mit modernen Grossteleskopen und fortschrittlicher Instrumentierung möglich geworden. Wegen der Bedeutung für die Entstehung und Evolution von Galaxien werden diese Arbeiten zukünftig weiter an Bedeutung gewinnen.<br /> <br /> In der vorliegenden Arbeit wurde die Integral-Field-Spektroskopie an zwei planetarischen Nebeln in der nächstgelegenen großen Spiralgalaxie M31 (NGC 224) getestet, deren Helligkeiten und Koordinaten aus einer Durchmusterung vorlagen. Hierzu wurden Beobachtungen mit dem MPFS-Instrument am russischen 6m - Teleskop in Selentschuk/Kaukasus sowie mit INTEGRAL/WYFFOS am englischen William-Herschel-Teleskop auf La Palma gewonnen. Ein überraschendes Ergebnis war, daß eins der beiden Objekte falsch klassifiziert wurde. Sowohl die meßbare räumliche Ausdehnung des Objektes als auch das spektrale Erscheinungsbild schlossen die Identität mit einem planetarischen Nebel aus. Mit hoher Wahrscheinlichkeit handelt es sich um einen Supernovaüberrest, zumal im Rahmen der Fehler an gleicher Stelle eine vom Röntgensatelliten ROSAT detektierte Röntgenquelle liegt.<br /> <br /> Die in diesem Projekt verwendeten Integral-Field-Instrumente wiesen zwei verschiedene Bauweisen auf, die sich miteinander vergleichen ließen. Ein Hauptkritikpunkt der verwendeten Instrumente war ihre geringe Lichtausbeute. Die gesammelten Erfahrung fanden Eingang in das Konzept des derzeit in Potsdam in der Fertigung befindlichen 3D-Instruments PMAS (Potsdamer Multi - Apertur - Spektrophotometer), welcher zunächst für das 3.5m-Teleskop des Calar - Alto - Observatoriums in Südspanien vorgesehen ist. Um die Effizienz dieses Instrumentes zu verbessern, wurde in dieser Arbeit die Kopplung der zum Bildrasterung verwendeten Optik zu den Lichtleitfasern im Labor untersucht. Die Untersuchungen zur Maximierung von Lichtausbeute und Stabilität zeigen, daß sich die Effizienz durch Auswahl einer geeigneten Koppelmethode um etwa 20 Prozent steigern lässt. / Popular scientific abstract: <br /> <br /> Currently there are two different approaches in the observational optical astronomy: On the first hand objects are imaged with cameras, on the other hand spectra are produced. The integral-field-technique is a relatively new way to combine both branches. The object image in the telecopes focus is sampled spatially and each spatial bin assigned to a spectrograph. Hence the object is not only sampled spatially but the spectral component is achieved as a third dimension, so the name 3D-Method. The result is like an image but each point consists of a whole spectrum. The use of this technique is to sample objects completely in contrast to standard slit spectroscopy. The strength of this method is to deal with high background light contamination. So the 3D method looks convincing for the new branch of extragalactic stellar astronomy with modern large telescopes. The detailled investigation of spatially resolved extragalactic populations in nearby galaxies is a rather new topic demanding newest telescopes and instrumentation. These investigations are very important in future to understand the origin and evolution of galaxies.<br /> <br /> In this thesis the Integral-Field-Spectroscopy was tested for two planetary nebulae in the Andromeda galaxy M31. Observations have been made using the MPFS on the Russian 6m telescope and the INTEGRAL/WYFFOS setup on the 4.2m WHT on La Palma. A surprising result was that one of the two objects was wrongly identified as a planetary nebula. The spatial extension and spectral details excluded this object class. With high probability this object is a supernova remnant.<br /> <br /> The integral-field-instruments used in this thesis had different technical layouts, which were to compare to each other. The main critics is the poor efficiency of both devices. The experience made was utilized to optimize the concept of the recently developed 3D-instrument PMAS (Potsdam Multi- Aperture Spectrophotometer). PMAS will be used for the 3.5 m telescope at the Calar Alto observatory. To improve efficiency, the coupling of the optical fibers used to sample the object and guide the light into the spectrograph was optimized in the laboratory. This investigations showed that an increase of the coupling efficiency by about 20 percent is possible by using immersion coupling between fibers and lenses.

The presented study was aimed to measurement of extracellular acid phosphomonoesterase activity in Norway spruce (Picea abies /L./ Karst.) pure stands. All study plots were chosen with respect to silvicultural treatment. Norway spruce mature stand 108-year-old (108); 36-year-old Norway spruce stand thinned from above (C3); 36-year-old Norway spruce stand thinned from below for (D4). Research was situated in Ecosystem station Rajec-Nemcice, the Czech Republic. The nature forest region of Drahanska Vrchovina upland (Drahansky bioregion) is ground area for Ecosystem Station Rajec-Nemcice, soil type was recognized as modal oligotrophic Cambisol (KAmd'). Typical is fir-beech forest vegetation zone. There are two major goals. As for soil activity of extracellular acid phosphomonoesterase, it was expressed from the view point of basal activity of the enzyme studied jointly with the response following the adding of the salicyl acid. As for roots, only the basal activity was measured by p-nitropohenol concentration. Presented data indicate negative effect of the salicylic acid adding into the enzyme activity.

The aim of the thesis on "Evaluation of the color of honey" was to determine why the instrument can not Lovibond Honey Colorpod 440110 to evaluate color of all types of honey and compare results of with spectrophotometric measurement methods. The work is divided into theoretical and practical part. The theoretical part is dedicated to honey, its composition and methods of evaluating color. The practical part describes the results obtained instrument Lovibond Honey Colorpod -- 440110, spectrophotometers Konica Minolta -- 3500d a FT -- NIR Antaris. In UV/VIS region not been demonstrated reason of errors of photometer. Maximum lightness had a very light honeys and the largest share of yellow had light honey. The highest values of absorbance attain dark honeys. In color space CIE L* a* b* had the largest share of red. Evaluation of the color of honey in the UV/VIS can not prove adulteration of honey always unambiguously, but there are signs by which honey can be described as suspicious. By analyzing the NIR spectrum is falsification of honey demonstrated using discriminant analysis.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Tereza Morávková Supervisor: Assoc. Prof. Přemysl Mladěnka, Pharm.D., Ph.D. Title of Thesis: Assessment of the complex stoichiometry of the active dexrazoxane metabolite ADR-925 with iron and copper by the standard Job'smethod Iron and copper are important trace elements which participate on many physiological processes in humans. Their kinetics in the organism is tightly regulated since both lack or excess of these elements are associated with pathological states. Free ions of iron and copper can catalyse reactive oxygen species (ROS) production and hence cause damage to proteins and DNA. Imbalance in these metals is linked with diabetes, cardiotoxicity, cirrhosis of the liver and neurodegenerative diseases as Alzheimer's disease or Parkinson's disease. Dexrazoxane is a drug with documented protecting effect against cardiotoxicity of anthracyclines. A former theory associated its protective effects against these cytotoxic drugs with the iron-chelating properties of its active metabolite ADR-925. The goal of this diploma thesis was to explore if ADR-925 is able to chelate Fe2+ , Fe3+ , Cu+ a Cu2+ ions at physiologically and pathophysiologically relevant pH values (4,5; 5,5; 6,8 and 7,5) and...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Lucia Hanuščinová Supervisor: Assoc. Prof. Přemysl Mladěnka, PharmDr., PhD. Title of diploma thesis: Ability of copper chelators to interact with iron and zinc Copper plays in the human organism a role of an element with indispensable significance, whose biological influence and effects depend on its quantity. With elevated concentrations in the human body, copper becomes toxic, resulting in pathological conditions. The most well-known diseases is the Wilson's disease, whose treatment consists of oral administration of chelators, i.e. chemical compounds, which are capable of binding copper ions in various proportions and eliminating them from the organism. Chelation therapy is currently the first choice after confirmation of the diagnosis. Chelation toxicity results from several factors, e.g. inhibition of copper dependent enzymes or low selectivity to metals. And precisely the selectivity of chelators is being discussed in this diploma thesis. An ideal chelator should not interact with any of the other physiological ions, that are necessary for the proper functioning of the organism. Five of the most frequently therapeutically or experimentally used substances /trientine, D-penicillamine,...

The purpose of this work was the establishment of the amount of chlorophyll in diverse species of deciduous trees in enviroment contaminated by road transport. To determine this, the tests were made from may to august on the leaves of a smallleaved lime tree (Tilia cordata) and sycamore tree (Platanus occidentalis).

A derivatization reaction, based on a copulation reaction of diazonium salt of an analyte with a reagent (1-aminonaphthalene or NEDA) producing colour, spectrophotometrically detectable product, has been used for determination of two analytes (procain hydrochloride, sulfamethoxazole) containing amino group in the molecule. The conditions of batch spectrophotometric determination were studied. It was found, that the batch determination is applicable only for procain hydrochloride analysis, because sulfamethoxazole gives time-unstable colour product. FIA arrangement was proposed and effects of selected parameters were optimized using two levels factorial design. The calibration dependences were measured for both analysis with both reagents, and subsequently were applied on determination of analytes in medical forms. Key words: diazotization, flow injection analysis, procaine, spectrophotometry, sulfamethoxazole