Iron transport in K562 cells: a kinetic study using native gel electrophoresis and 59Fe autoradiography

AbstractThe exact mechanisms of iron transport from endosomes to the target iron containing cellular proteins are currently unknown. To investigate this problem, we used the gradient gel electrophoresis and the sensitive detection of 59Fe by autoradiography to detect separate cellular iron compounds and their iron kinetics. Cells of human leukemic line K562 were labeled with [59Fe]transferrin for 30–600 s and cellular iron compounds in cell lysates were analyzed by native electrophoretic separation followed by 59Fe autoradiography. Starting with the first 30 s of iron uptake, iron was detectable in a large membrane bound protein complex (Band I) and in ferritin. Significant amounts of iron were also found in labile iron compound(s) with the molecular weight larger than 5000 as judged by ultrafiltration. Iron kinetics in these compartments was studied. Band I was the only compound with the kinetic properties of an intermediate. Transferrin, transferrin receptor and additional proteins of the approximate molecular weights of 130 000, 66 000 and 49 000 were found to be present in Band I. The labile iron compounds and ferritin behaved kinetically as end products. No evidence for low molecular weight transport intermediates was found. These results suggest that intracellular iron transport is highly compartmentalized, that iron released from endosomal transferrin passes to its cellular targets in a direct contact with the endosomal membrane complex assigned as Band I. The nature of the labile iron pool and its susceptibility to iron chelation is discussed

Analysis of the Highest and Best Use of a Nursery School Building in Plzeň

Diplomová práce se věnuje analýze nejvyššího a nejlepšího využití. První částí práce je část teoretická, která se zabývá především metodami ocenění a dále podstatou analýzy. Tato analýza je aplikována na objekt školky v Plzni, kde se zkoumá, zda navržené varianty vyhoví čtyřem zkouškám. Výsledkem je zjištění nejvyššího a nejlepšího využití objektu.This diploma thesis deals with analysis of the highest and best use. The first part of the thesis is the theoretical part, which deals mainly with valuation methods and the essence of analysis. This analysis is applied to the Nursery School in Pilsen, where it examines whether the proposed variants meet the four tests. The result is finding the highest and best use of the object.

Boron Doped Nanocrystalline Diamond Films for Biosensing Applications

With the rise of antibiotic resistance of pathogenic bacteria there is an increased demand for monitoring the functionality of bacteria membranes, the disruption of which can be induced by peptide-lipid interactions. In this work we attempt to construct and disrupt supported lipid membranes (SLB) on boron doped nanocrystalline diamond (B-NCD). Electrochemical Impedance Spectroscopy (EIS) was used to study in situ changes related to lipid membrane formation and disruption by peptide-induced interactions. The observed impedance changes were minimal for oxidized B-NCD samples, but were still detectable in the low frequency part of the spectra. The sensitivity for the detection of membrane formation and disruption was significantly higher for hydrogenated B-NCD surfaces. Data modeling indicates large changes in the electrical charge when an electrical double layer is formed at the B-NCD/SLB interface, governed by ion absorption. By contrast, for oxidized B-NCD surfaces, these changes are negligible indicating little or no change in the surface band bending profile

OPTIMAL ADDENDUM COEFFICIENTS OF CYLINDRICAL INVOLUTE GEARING FROM POINT OF VIEW ITS EFFICIENCY

The optimisation of gearboxes, namely of teethed wheels is still valid subject of scientific research. The development of software products for multiparametrical optimisation at Machine Engineering Faculty University of Technology Bratislava was concluded into successful solutions of arbitrary defined objective functions. In the article are given the results of teeth gears optimisation with involute gearing from point of view of its efficiency. The calculation included the complete geometrical and strength check of gearing according to DIN 3990 level B / ISO 6336 as the restriction conditions in original way.The optimisation of gearboxes, namely of teethed wheels is still valid subject of scientific research. The development of software products for multiparametrical optimisation at Machine Engineering Faculty University of Technology Bratislava was concluded into successful solutions of arbitrary defined objective functions. In the article are given the results of teeth gears optimisation with involute gearing from point of view of its efficiency. The calculation included the complete geometrical and strength check of gearing according to DIN 3990 level B / ISO 6336 as the restriction conditions in original way

The impact of including rest time between transport and slaughter on the IGF-1 and cortisol concentration in the blood serum of barrows

The aim of our experiment was to observe the differences of IGF-I and cortisol concentrations in the blood serum of pigs when killed immediately after transport and after a 48 hour stay in a slaughterhouse.  We tested 24 Large white barrows from one breed with the average weight of 105 kg. The first group went through a 2 hour long transport before slaughter. After being delivered to the slaughterhouse, the barrows were killed immediately. After transport, the second group was stabled for 48 hours in a stabling space near the slaughterhouse and the barrows were killed after that. There were 12 animals in each experimental group. Blood samples were taken immediately after slaughter. We used the ELISA method to determine IGF-I and cortisol concentration in the serum. The IGF-I concentration in the serum was significantly lower (p < 0.01) in the barrow group that was killed after the 48 hour stay in the stabling space near the slaughterhouse, as opposed to the barrow group that was killed immediately after transport. The cortisol concentration in the serum of both experimental groups did not display significant differences. Our results indicate that stabling animals in a waiting stabling space near the slaughterhouse can cause a long-lasting stress instead of relax

Characterization of protamine influence on indirect electrophoretic determination of heparin

Cílem této bakalářské práce bylo studium interakcí heparinu a protaminu v prostředí kapilární zónové elektroforézy pomocí jejich vlivu na velikost plochy píku tetraargininu. Byla použita křemenná kapilára o vnitřním průměru 50 µm a celkové délce 50 cm. Efektivní délka kapiláry byla 41,5 cm a vzorek byl dávkován na dlouhý konec kapiláry. Byl optimalizován způsob dávkování jednotlivých zón, vhodné složení základního elektrolytu bylo již známo z předešlých experimentů, na které tato práce navazovala. Základní elektrolyt byl složen z 0,1 M roztoku kyseliny trihydrogenfosforečné a 0,1% roztoku hydroxyethylcelulózy. Vzorek byl dávkován hydrodynamicky tlakem 5,0 kPa po dobu 120 s. Následně byla stejným tlakem po dobu 90 s dávkována zóna základního elektrolytu. Poslední dávkovanou zónou byla zóna tetraargininu o koncentraci 1 mg/ml, která byla dávkována hydrodynamicky tlakem 5,0 kPa po dobu 3 s. Posledním krokem bylo vložení separačního napětí 30 kV po dobu 30 s. Tetraarginin byl detegován při λ = 200 nm. Byly změřeny vlivy samotného heparinu/ protaminu a jejich směsí v různých vzájemných poměrech. Bylo ověřeno, že se zvyšující se koncentrací heparinu dochází k poklesu plochy píku tetraargininu. Nově bylo zjištěno, jaký vliv na velikost tohoto účinku má přítomnost protaminu. Zde došlo k překvapivému...Katedra analytické chemieDepartment of Analytical ChemistryFaculty of SciencePřírodovědecká fakult

THE INFLUENCE OF FLOOR QUALITY ON DAIRY COW BEHAVIOUR AND THEIR BODY CLEANLINESS

The aim of this study was to evaluate the influence of floor quality on the behaviour and cleanliness of the diary cow‘s body. The analysis results show that cows housed in the section with the separated manure solids spent more time lying (41.47%) and also laid less in the corridor manure (0.91%) than the cows in the section with separated manure solids and the rubber mats where they were laying generally less (35.71%), they spent more time lying in the corridor manure (1.55%) and in terms of preference of lying area the cows clearly preferred the separated manure solids (25.8%). Based on observations we can say that cows housed in sections of slurry solid phase were cleaner (average 6.13 points) than cows in the section with the rubber and a separated manure (averagely 7.28 points). Nowadays the free housing systems with different types of litter and boxes are frequently used, while the proper littering influences the lying preferences of an animal. In practice, for the dairy cows the deepened lying boxes littered with natural materials such as sawdust, shavings, straw, dried manure and sand boxes or elevated boxes covered with a synthetic material, mattresses or mats are used

Nitrogen saturation of tissuesNitrogen saturation of tissues

Cílem této semestrální práce bylo vytvořit aplikaci pro OS Android, která umožňuje naplánovat ponor s dýchacím přístrojem. Ve výpočtech je jako dýchací směs brán vzduch. Aplikace umožňuje zobrazit grafu ponoru včetně dekompresních zastávek a zobrazit dekompresní tabulky.The goal of this semestral thesis was to create an Android application able to plan a dive with a breathing apparatus. In the calculations air is used as the breathing mixture. The aplication is able to draw the dive graph and display the decompression tables.