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Bioactive peptides as a component of anti-aging cosmetics
Jatzová, Katarína ; Obruča, Stanislav (referee) ; Muthný,, Tomáš (advisor)
Ageing is a natural part of every human life cycle. During ageing there are lots of changes in the organism. One of the main pillars of the cosmetics industry is the development of active compounds that are fighting signs of skin ageing. The components as bioactive peptides are considered to be promising anti-ageing products, mainly because of the possibility to precisely define their chemical structure and therefore achieve more effective biological targeting. One of the signs of skin ageing is the weakening of the connections between epidermal cells and the extracellular matrix, decreasing expression of adhesion molecules and molecular components of dermo-epidermal connection. The most abundant adhesive receptors in the skin are integrins. Their ligands are extracellular matrix molecules, e. g. laminin or fibronectin. The minimal recognition sequence of integrins receptors is the amino acid motif arginine- glycine- aspartic acid (RGD). This sequence is also very interesting in terms of cosmetic applications because it provides the ability to create new and effective bioactive peptides. The subject of present work is basic safety testing of four peptides with RGD motif. A sequence of three peptides was modified by addition of glycine amino acids, or alanine. The expected effect was to improve interaction with integrin receptors. In each of the sample, levels of endotoxin was determined in order to exclude any possible interfering effects on the viability of cells. Subsequently, in NIH3T3 mouse fibroblasts viability was monitored by MTT assay and morphology. The quantity of obtained protein had been determined to increase data interpretation relevance.
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Synthesis and Study of Nano-Structured Perovskites for Applications in Organic Electronics
Jančík Procházková, Anna ; Kuřitka, Ivo (referee) ; Částková, Klára (referee) ; Krajčovič, Jozef (advisor)
Nanočástice perovskitů halogenidů kovů vykazují unikátní vlastnosti, především výjimečně vysoké hodnoty kvantových výtěžků fluorescence, které předurčují tyto materiály pro aplikace v optoelektronických a fotonických zařízeních. Tato práce popisuje přípravu nanočástic perovskitů halogenidů kovů pomocí stabilizačních činidel inspirovaných přírodou. Stabilizační činidla zde slouží nejen ke stabilizaci, ale i k modifikaci povrchu nanočástic za účelem zvýšení funkčnosti výsledných nanostruktur. Úvod práce popisuje optimalizaci přípravy nanočástic precipitační technikou za použití stabilizačních činidel; jako stabilizační činidlo byl zvolen adamantan-1-amin spolu s hexanovou kyselinou. Bylo prokázáno, že klíčový vliv na optické vlastnosti výsledných koloidních roztoků má volba rozpouštědel a teploty při precipitaci. Mimo jiné byl zkoumán vliv koncentrace prekurzorů na výslednou morfologii a optické vlastnosti nanočástic a jejich koloidních roztoků. V neposlední řadě byly nanočástice stabilizovány adamantan-1-aminem spolu s různými karboxylovými kyselinami a byly studovány optické vlastnosti a koloidní stabilita výsledných koloidních roztoků. V dalším kroku byly nanočástice perovskitů stabilizovány pomocí proetogenních aminokyselin L-lysinu and L-argininu. Takto stabilizované nanočástice vykazovaly úzká emisní spektra ve viditelné oblasti a kvantové výtěžky fluorescence dosahující hodnot téměř 100 %. Stabilizace nanočástic prostřednictvím postranních skupin aminokyselin byla prokázána navázáním chránící terc-butoxykarbonylové skupiny na -amino skupinu. Nanočástice stabilizované modifikovaným lysinem v průběhu jejich přípravy vykazovaly závislost optických vlastností na přítomnosti vody. Předpokládá se, že molekuly vody jsou schopné kontrolovat růst krystalové mřížky po navázání na prekurzory perovskitů a ovlivňovat tak výslednou velikost nanočástic, což vede k projevení kvantových jevů. Spojení nanočástic perovskitů s peptidy představuje nový typ materiálů kombinujících výjimečné optické vlastnosti se samoorganizačními a senzorickými vlastnostmi. Tento koncept byl představen přípravou nanočástic perovskitů stabilizovaných cyklo(RGDFK) pentapeptidem. Vzhledem k citlivosti peptidů na jejich byly nanočástice stabilizovány peptidovými nukleovými kyselinami, robustními analogy nukleových kyselin. Ke stabilizaci nanočástic byl připraven monomer a trimer peptidové nukleové kyseliny obsahující thymin jako dusíkatou bázi. Thymin byl na povrchu nanočástic dostupný k interakci s adeninem přes vodíkové můstky umožňující přenos náboje. Kombinace peptidových nukleových kyselin a perovskitů s unikátními optickými vlastnostmi otevírá aplikační možnosti zejména v oblasti optických senzorů.
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Characterization of organic nanoparticles with encapsulated antimicrobial peptides
Vejrostová, Petra ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on characterization of particles containing encapsulated antimicrobial enzyme lysozyme. The theoretical part deals with characterization of antimicrobial peptides and their description. Further part of review was focused on lysozyme, the selected representative used in this thesis, its structure, mechanism of action and possible usage. In the experimental part the Hartree-Lowry method was used for lysozyme detection, determination of encapsulation efficiency and for detecting the amount of lysozyme released after incubation in model physiological environment and in model foods. In process of encapsulation the highest amount of lysozyme was packed into 1% chitosan particles, manually prepared alginate particles and into liposomes. During study of stability of particles in model foods as the least stable manually prepared chitosan particles were found. The released lysozyme exhibited changes probably caused by its degradation. The highest influence on particles proved 3% acetic acid. During studying the stability of the particles in artificial digestive fluids as the most unstable manually prepared chitosan particles were found, while alginate particles were the most stable. The thesis also deals with changes in antimicrobial activity of encapsulated lysozyme in prepared particles and after its application into the model environments. A gram-positive bacteria Bacillus subtilis was used in order to test the antimicrobial activity. Antimicrobial tests showed that after encapsulation antimicrobial activity of lysozyme was substantially decreased in most samples. Size and stability of prepared particles was tested using dynamic light scattering.
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Proteomics for beginners
Hubálek, Martin ; Vrbacký, M. ; Křenková, Alena
Proteomic tutorial for the beginners will summarise and present current methods to analyse and evaluate proteomic samples. The tutorial will be divided into two parts. The first part will focus on data accquisition process and interpretation of spectra. The second part will focus on protein quantification.
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Characterization of new HILIC stationary phases
Vaclová, Jana
Hydrophilic interaction liquid chromatography is a frequently used separation method for analysis of polar compounds. It is an alternative method to reversed-phase chromatography, where these compounds show insufficient or very weak retention. A high number of stationary phases are currently available for HILIC and new ones are still being developed. The aim of this diploma thesis was to characterize and compare three relatively new HILIC columns containing an unmodified silica gel (HILIC-A), aminopropyl modified silica (HILIC-B) and polyhydroxyl chain modified silica (HILIC-N) as stationary phase. Based on the study of the effect of acetonitrile content in mobile phase on the retention of a model set of peptides, a multimodal retention mechanism was demonstrated. Analysis of 18 model analytes with different pKa values showed, that the composition of aqueous part of mobile phase (buffer), more specifically it's concentration and pH value, has a significant impact on retention of ionized analytes and peptides on the studied stationary phases. A significant contribution of ionic interactions to retention was observed on HILIC-B and HILIC-A columns. The retention of basic compounds on the HILIC-B column increased with increasing ionic strength, while it decreased on the HILIC-A column. Increasing the...
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Determination of structure and dynamics of biomolecules by theoretical calculations of NMR spectroscopic parameters
Benda, Ladislav ; Sychrovský, Vladimír (advisor) ; Munzarová, Markéta (referee) ; Hrabal, Richard (referee)
iv Abstract Subject: Determination of structure and dynamics of biomolecules by theoretical calcu- lations of NMR spectroscopic parameters Author: Ladislav Benda, ladislav.benda@gmail.com Department/Institute: Institute of Organic Chemistry and Biochemistry, AS CR Supervisor: Dr. Vladim'ır Sychrovsk'y, Institute of Organic Chemistry and Biochemistry, AS CR, vladimir.sychrovsky@uochb.cas.cz Abstract: This doctoral work was focused on theoretical modeling of nuclear magnetic resonance (NMR) parameters in peptides and nucleic acids. Dependences of NMR para- meters on molecular structure and solvation were primarily modeled. Great emphasis was put on the comparison of the calculated data with the NMR experiment. The molecular models studied included the l-alanyl-l-alanine di-peptide (AA) and the phosphate group of nucleic acid backbone. Conformations of all three charged forms of AA in solution were determined and the respective pH-induced changes of experimental NMR chemical shifts and nuclear spin-spin coupling constants were explained. Dependences of NMR cross-correlated relaxation rates on the AA backbone geometry were calibrated. The 31 P NMR parameters in nucleic acid phosphate were systematically calculated in dependence on the backbone conformation and the phosphate solvation pattern. Qualitative rules...
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Influence of compounds produced by cyanobacteria on coagulation of humic waters
Brabenec, Tomáš ; Pivokonský, Martin (advisor) ; Benešová, Libuše (referee)
The diploma thesis investigates the coagulation of nature waters with humic substances and peptides/proteins produced by Microcystis aeruginosa during water treatment process with aluminum coagulants. It was confirmed that the efficiency of coagulation mechanism of humic substances and peptides/proteins strongly depends on the pH value, because pH value is limited factor of a charge properties of peptides/proteins, humic substances and hyrolysis products of coagulants. The optimal pH range for the coagulation of humic substances was 5- 6 and for cyanobacterial peptides/proteins 5-6.5. In both cases a charge neutralisation and an adsorption were dominant mechanism of coagulation. Optimal pH range for the coagulation of humic substances in the presence of cyanobacterial peptides/proteins was identical as well as optimal pH range of humic substances with absence of peptides/proteins (5-6) but the presence of peptides/proteins positively influenced particle coagulation of humic substances, because the optimal concentration of the coagulants decreased to more than half. An interaction between humic substances and peptides/proteins was demonstrated. A positive effect of coagulation was observed in the case of jar tests with absence of coagulants at a very low pH range (<4). Keywords Humic substances; AOM...
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Modeling of bio-inorganic interfaces
Trachta, Michal ; Bludský, Ota (advisor) ; Nachtigall, Petr (referee)
Dynamic atomistic description of bio-inorganic interfaces represents a challenging problem for contemporary computational chemistry. A detailed analysis of processes occurring on the interface between biomolecule and inorganic material can help our understanding of various processes, ranging from chromatography and protein separation to protein immobilization techniques and their effect on enzyme activity or protein conformational stability. High complexity of bio- inorganic interfaces prevents detailed investigation using accurate, but computationally demanding ab initio methods. Since reliable empirical potentials are not available for these systems, the aim of this work is to develop force fields based on ab initio data as well as a general methodology for parameterization of such force fields. Our potential fitting procedure was carried out in an automated fashion based on molecular dynamics simulation. The resulting potentials were applied for investigation of inorganic material's influence on polypeptide conformations.
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Characterization of extracellular and intracellular organic matter produced by phytoplankton in relation to water treatment
Zezulová, Tereza ; Pivokonský, Martin (advisor) ; Janda, Václav (referee)
This study investigated characteristics of algal organic matter (AOM) derived from four species (cyanobacteria Microcystis aeruginosa and Merismopedia tenuissima, diatom Fragilaria crotonensis and green alga Chlamydomonas geitleri) dominating phytoplanktonic populations in the reservoirs supplying drinking water treatment plants - the water reservoirs Švihov (the Želivka water treatment plant) and Josefův Důl (the Jizerské Mountains), Czech Republic. The four mentioned microorganisms were cultivated under laboratory conditions and the algal growth was quantified by a number of cells, chrorophyll-a and optical density. Production of AOM was evaluated by dissolved organic carbon concentration measurements, by specific UV absorbance, the amount of peptide/protein and non-peptide (carbohydrate) contents, water affinity and molecular weight (MW). AOM of all considered microorganisms are formed predominantly by hydrophilic substances with low values of specific UV absorbance (< 2 L/(m.mg)). Considerable portions of both types of AOM, i.e. extracellular and cellular organic matters are represented by substances with MW < 1 kDa or with MW > 100 kDa. However, the total amount and composition of AOM significantly depend on algal species and their growth. The obtained results imply that the knowledge of AOM...
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