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Study of nucleic acids by means of drop coating deposition Raman microspectroscopy (DCDR)
Souček, Pavel ; Kopecký, Vladimír (advisor) ; Mojzeš, Peter (referee)
The work was focused on possibilities of the new technique of nonresonance Raman spectroscopy - drop coating deposition Raman (DCDR) spectroscopy upon study of nucleic acids. DCDR spectroscopy is based on deposition of a small droplet of the studied sample on a hydrophobic surface, where after evaporation of the solvent, ring of the studied material in glass phase is formed. That way the material is concentrated and upon measurements by means of Raman microspectrometry the increase of signal of several orders of magnitude is observed with respect to the standard technique of sample measurements in solvent. In this work we studied behavior of DNA 12-mer and DNA with 3000 base pairs. Our research showed that DCDR spectra of DNA dissolved in deionized water can be measured up to concentration of 30 M per base. The DCDR spectra are similar to those measured from solvents. Nevertheless, the deposited sample never forms a ring. It was shown that the size of nucleic acids didn't play dominant role upon ring formation but their charge which lead to repulsion of the molecules. After adding of sodium ions and subsequently also magnesium ions the ring formation was observed. Factor analysis of spectral maps demonstrated that formed rings are not fully homogenous and that upon their origin condensation of DNA could...
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Spectroscopic study of interaction of antimicrobial peptides with model membranes
Tesař, Adam ; Kopecký, Vladimír (advisor) ; Kočišová, Eva (referee)
Antimicrobial peptides (AMP) have a great potential in medicine and pharmacy. Mechanism of their impact is an interaction with a cell membrane leading to the penetration of the membrane. The way of disruption of the cell membrane is not completely understand, therefore we focused on the interaction of AMP HAL-1, isolated from the venom of the bee Halictus sexctinctus, with a model membrane of 100 nm liposomes consisting of phosphatidylcholin and phosphatidylglycerol. Circular dichroism and infrared spectroscopy (FTIR) proved the change of the secondary structure from the random coil of free HAL-1 to -helix in an interaction with the membrane. The next step was preparation of the lipid bilayer on the surface of ATR prism, which will enable usage of the polarized FTIR spectroscopy to study the interaction of AMP with model membranes in future. Therefore, the ATR-FTIR spectroscopy and factor analyses were applied to study dynamics of drying of the liposomes and their subsequent hydration also with an addition of HAL-1. We focused on the stabilization of the system. Hydratation of the lipid bilayer by 2 µl sample showed stability for minutes, nevertheless after dilution the stabilization decay in minutes. The protective influence of the peptide on the lipid bilayer and slowing down of the drying out of the...
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Diagnostics of neurodegenerative diseases by means of Raman spectroscopy
Klener, Jakub ; Kopecký, Vladimír (advisor) ; Matějka, Pavel (referee)
Therapies of neurodegenerative diseases are often very difficult and their success depend on an early diagnose. From that reason we have been developing new diagnostic method for multiple sclerosis and Alzheimer disease by drop coating deposition Raman (DCDR) spectroscopy of cerebrospinal fluid (CSF) in this work. We found out conditions of measurements, where spectra were reproducible and accepted for standard diagnostic practices. We discovered that CSF has fast degradation at a room temperature, which was detectable in spectra after 5 hours, and degradation due to refreezing. DCDR spectra of CSF from individual patients were analyzed by factor and cluster analysis. Multiple sclerosis was manifested by lower intensity of a Raman band at 1080 cm−1 , which is probably connected with more general pathologic state. Spectral changes caused by Alzeheimer disease were more complex and beside changes mentioned above also changes connected with composition and conformation of proteins were identified in regions 1200-1800 cm−1 and 2870-2950 cm−1 . Additionally, we succeeded in distinguishing of young healthy patients from older patients in DCDR spectra. In this work were checked up, that DCDR is good diagnostic method for clinical practices for determining neurodegenerative diseases through the complex...
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Comparison of climate change policy of the United States and People's Republic of China between 2004 and 2009
Kopecký, Václav ; Kozák, Kryštof (advisor) ; Sehnálková, Jana (referee)
Bachelor thesis "Comparison of climate change policy of the United States and People's Republic of China between 2004 and 2009" deals with a development, main actors and key documents of two above mentioned countries between years 2004 and 2009. Contrary to different social-economic conditions, political system or rhetoric, the two governments had, surprisingly, a lot of in common in their stance towards climate change or policy of mitigation. They were unwilling to curb their GHG emissions mandatorily but they supported renewable resources of energy, improvements of energy intensity in industry, improvements of energy efficiency and implementations of
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Protein dynamics studied by means of optical spectroscopy
Pazderka, Tomáš ; Kopecký, Vladimír (advisor) ; Maloň, Petr (referee)
In this diploma thesis a software for Two-dimensional correlation spectroscopy (2DCoS) was programmed and tested. Possible spectral phenomena in 2DCoS analysis of vibrational circular dichroism were tested on model cases. Thermal and time dependent series of Raman and Raman optical aktivity spectra of lysozyme protein were measured for practical applications of 2DCoS. Fibrillation and denaturation process of lysozyme was analyzed by means of 2DCoS and Factor analysis (FA). Proteins NKR_P1A and NKR_P1C were also studied and their thermal dynamics was monitored by means of Fourier transformed infrared spectroscopy and Raman spectroscopy. Dynamical behaviour of all studied proteins was characterized using 2DCoS.
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