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Improved determination of DNA duplex stability by using UV spectroscopy
Víšová, Ivana ; Štěpánek, Josef (advisor) ; Kočišová, Eva (referee)
This work deals with determination of thermodynamic parameters concerning the temperature induced decomposition of DNA duplexes. Precise temperature dependences of solution UV absorption spectra have been measured for several concentrations. Proposals and tests of various data analysis procedures, with aim to find out optimal way how to determine the thermodynamic potentials changes, represent the core of the work. It has been demonstrated that the combination of a factor analysis and a fit of thermodynamic equations to temperature dependences of obtained coefficient values, enables these parameters to be obtained from a measurement performed for only one concentration and with better precision than by using the so far recommended method based on a melting temperature determination for several concentrations.
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Study of liposomes by using of drop coating deposition Raman spectroscopy
Vodáková, Adéla ; Kočišová, Eva (advisor) ; Bednárová, Lucie (referee)
in english: Goal of the Bachelor Thesis was an optimization of liposome membrane preparation with different composition for the drop coating deposition Raman (DCDR) method, measuring of the DCDR spectra from dried droplets and their interpretation. The DCDR method lies in deposition of small volume of suspension (~μl) on special surface and measuring of Raman spectra from the ring formed at the edge of the dried droplet. The main advantage of used method is the possibility of measuring spectra from the sample with low concentration. Our results show that the DCDR method is very useful in studies of biological membranes with different composition. Two synthetic lipids and one natural extract were chosen for making a comparison. We have found out that spectra obtained from the ring of dried droplet are not different from the spectra measured in the suspension. The important characteristic of spectra is that dried droplet keeps the same lipid phase as in suspension. Spectral mapping proved good reproducibility of the signal inside the ring.
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Microcalorimetric determination of RNA triplex thermodynamic stability
Říhová, Martina ; Štěpánek, Josef (advisor) ; Kočišová, Eva (referee)
Nazcv prace: Mikrokalorimelricke slanovcni termodynamicke stability triplexu RNA Autor: Martina Rihova Kaledra (ustav): Fyzikalni ustav UK Vcdouci bakalafske prace: Prof. RNDr. Josef Stcpanck, Csc. e-mail vedouciho: stepjosiyj;karlov.mff.cuni.cz Abstrakt: Prace je zamC'fena na urcovani termodynamickych charaktcristik duplexu a triplexu ribonukleove kyscliny. Rozpad a v/nik komplexu hyl sledovan na diferencnim skcnovacim mikrokalorimelru. Jako vzorky byly pouzity smesne ro/toky syntelicky pfipravenych homopolymerii RNA, polyA a polyU, o ruznych stcchiometrickych pomerech. Z namcfenych dal byly urceny termodynamickc paramclry komplexu, tcdy odpovidajici zmeny entalpie AH a entropic AS a teplola tani komplexu. Vysledky ukazaly, /e pfi techto mcfcnich dochazi k systematickc /mene namerenych charakteristik pfi opakovanych scanech. Tento jcv patrnc souvisi s intcrakci RNA s povrchem mefici kyvcty a jc tedy zavazny z hlediska aplikovatclnosli scanovaci mikrokalorimctric pro pfesna kvantitativni tennodynamicka studia ribonukleove kyscliny. Zmcny teploty tani zpusobenc pridanim hofciku do rozloku uka/aly, zc jeho ucinkem so pfi pfipojeni ifctiho vlakna k duplexu RNA komplex stabilizuje. Klicova slova: triplex RNA, duplex RNA, mikrokalorimetric, termodynamickc charakteristiky, teplota tani. Title: Microcalorimetric...
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Study of oligonucleotide penetration through liposomal membrane
Nováková, Jana ; Kočišová, Eva (advisor) ; Holoubek, Aleš (referee)
Modified oligonucleotides, short pieces of nucleic acids, have become potential candidates for treatment of viral, malignant or genetic diseases. Efficient application of modified oligonucleotides requires their sufficient penetration inside the cell through the lipid membrane. Diploma thesis employs liposomes, synthetic vesicles formed by the lipid bilayer, as a model system for the study of interaction of lipid and complex of homooligonucleotides dT15 with cationic porphyrin CuTMPyP4 using Raman spectroscopy. Raman spectra were measured in solution on the slide and using special technique of drop coating deposition Raman spectroscopy (DCDR) on integrated Raman confocal microspectrophotometer. Processes of liposome formation and sample preparation were optimized. Experimental data were analysed using factor analysis based on singular value decomposition (SVD) algorithm. It was observed that interaction of oligonucleotide with porphyrin induces fluorescent background increase. Raman spectra of complex of oligonucleotide with porphyrin exhibit the domination of porphyrin vibrational bands. Stretching vibrations of lipid assigned to hydrogens of fatty acids are observed in the spectra of complex with liposomes. The observed spectral changes show the interaction of lipid with complex of oligonucleotide and...
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Study of angular and time characteristics of suspensions by using of elastic scattering
Peikertová, Pavlína ; Kočišová, Eva (advisor) ; Holoubek, Aleš (referee)
Elastic light scattering represents relatively simple and noninvasive technique to study angular dependencies of the scattered light intensity that contains important information about scattering objects. In the frame of this thesis the data from adapted computer controlled apparatus Photo-gonio-diffusometre type 42.000 were collected. Experimental set-up was upgraded to use it for small sample volumes. Two model systems were studied: polymer and latex suspensions and real systems: liposomal suspension of various sizes, as well. The size estimation by using of Guinier plot was performed from experimental data. This estimation is in good agreement with the defined sizes in case of model systems. For suspensions of liposomes the results are also in relatively good agreement with expected values. Aggregation process and subsequent sedimentation after addition of porphyrin CuTMPyP4 was observed for silver borohydrid colloid. Acquired time and angular dependencies were processed by factor analysis that is able to make a qualitative and temporal separation of both processes
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The amount of inforamtion in the information society and its social consequences
Kočišová, Eva ; Lupač, Petr (referee) ; Duffková, Jana (advisor)
This bachelor thesis presents the problematic of quantitative growth of information volume in the contemporary developed societies (with regard to the historical development). It observes the influence of this growth in some areas of the society (particularly media communication, democracy, science and education) and also in a wider social context. It also examines the change of processing information (and its related behaviour) as a form of adaptation to the large information growth.
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