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The design of ultrasensitive immunosensors based on a new multi-signal amplification gold nanoparticles-dotted 4-nitrophenylazo functionalised graphene sensing platform for the determination of deoxynivalenolSunday, Christopher Edozie January 2014 (has links)
Philosophiae Doctor - PhD / A highly dispersive gold nanoparticle-dotted 4-nitrophenylazo functionalised graphene
nanocomposite (AuNp/G/PhNO2) was successfully synthesised and applied in enhancing
sensing platform signals. Three label-free electrochemical immunosensors for the detection of deoxynivalenol mycotoxin (DON) based on the systematic modification of glassy carbon electrodes (GCE) with AuNp/G/PhNO2 was effectively achieved. General electrochemical impedance method was employed for the sensitive and selective detection of DON in standard solutions and reference material samples. A significant increase in charge transfer resistance (Rct) of the sensing interface was observed due to the formation of insulating immune-complexes by the binding of deoxynivalenol antibody (DONab) and deoxynivalenol antigen (DONag). Further attachments of DONab and DONag resulted in increases in the obtained Rct values, and the increases were linearly proportional to the concentration of DONag. The three immunosensors denoted as GCE/PDMA/AuNp/G/PhNH2/DONab, GCE/Nafion/[Ru(bpy)3]2+/AuNp/G/PhNH2/DONab and GCE/Nafion/[Ru(bpy)3]2+/G/PhNH2/DONab have detection range of 6 – 30 ng/mL for DONag in standard samples. Their sensitivity and detection limits were 43.45 ΩL/ng and 1.1 pg/L; 32.14ΩL/ng and 0.3 pg/L; 9.412 ΩL/ng and 1.1 pg/L respectively. This result was better than those reported in the literature and compares reasonably with Enzyme Linked Immunosorbent Assay (ELISA) results. The present sensing methodology represents an attractive alternative to the existing methods for the detection of deoxynivalenol mycotoxin and other big biomolecules of interest due to its simplicity, stability, sensitivity, reproducibility, selectivity, and inexpensive instrumentation. And they could be used to develop high-performance, ultra-sensitive electrochemiluminescence, voltammetric or amperometric sensors as well.
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Transformações na matéria orgânica nos sedimentos e águas eutrofizados da Represa Ibirité (Ibirité-Sarzedo, MG) tratados com nitrato de cálcio em microcosmosColzato, Marina 19 November 2010 (has links)
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Previous issue date: 2010-11-19 / Financiadora de Estudos e Projetos / Due to the growing concern with the environment and the welfare of future generations the search for the restoration of degraded areas and the reestablishment of equilibrium between the anthropogenic and natural environments are nowadays highlighted. The Ibirité Reservoir (Ibirité-Sarzedo, MG) is an example of water body impacted by the disorganized urbanization and strong industrial development in the metropolitan Belo Horizonte, MG, SE Brazil. The reservoir, built by the REGAP-Petrobras in the sixties to supply water to its industrial processes, is a eutrophic water body due to the continuous contribution of urban sewage (external loading) and the consequent internal loading of nutrients (mainly of phosphorus), the primary element responsible for the eutrophication process. Among the existing technologies for the treatment of eutrophic sediment focusing on the P immobilization is the artificial supply of nitrate as an electron acceptor to the stimulation of the natural denitrification process. In this stimulation process, the different geochemical forms of reduced iron are oxidized to iron (III) oxi-hidroxides that adsorbs the phosphorus present in the sediment pore water. In addition to that, the metallic sulfides and the organic matter are oxidized as well. This work aimed at the development of laboratory experiments in a bench scale (microcosms) in which sediments and water of the Ibirité reservoir were treated with nitrate addition in order to evaluate the organic matter transformations from caused this treatment. The results indicate that there was, in the treatment microcosms, an increase in the redox potential, an almost complete abatement of the acid volatile sulfide (AVS) of the sediments, and a significant reduction of available phosphorus in the water. The UVvis spectroscopy indicated differences in the spectral profiles between the control and treatment microcosms, but uniform trends along the experiment were not identified. The elemental analysis showed agreement with the results of AVS and nitrogen species in sediment pore water data, with the reduction of sulphur and nitrogen contents. / Diante da crescente preocupação com o meio ambiente e o bem estar das futuras gerações, destacam-se cada vez mais a busca pela recuperação das áreas degradadas e o restabelecimento do equilíbrio nas relações entre os meios natural e antrópico. A Represa Ibirité (Ibirité-Sarzedo, MG) é um exemplo de corpo de água impactado pela urbanização desorganizada e crescimento industrial da região metropolitana de Belo Horizonte, MG. A represa, construída pela REGAP-Petrobrás na década de 60 para suprimento de água em seus processos industriais, encontrase eutrofizada devido ao contínuo aporte de esgotos urbanos não tratados (carga externa) e à conseqüente carga interna de nutrientes, principalmente o fósforo, o principal responsável pelo processo de eutrofização. Dentre as tecnologias existentes para o tratamento de sedimentos eutrofizados, visando a imobilização do P, figura a adição de nitrato como um receptor de elétrons para o estímulo do processo natural de desnitrificação. Nesta estimulação, as diferentes formas geoquímicas de ferro reduzido são oxidadas a óxi-hidróxido de ferro (III) que adsorve o fósforo presente nas águas intersticiais, promovendo também, a oxidação dos sulfetos metálicos e a matéria orgânica presentes nos sedimentos. Neste trabalho o tratamento dos sedimentos e águas eutrofizados da Represa Ibirité foi avaliado em escala de bancada, utilizando microcosmos, onde se estudou a transformação da matéria orgânica presente nas águas de coluna, nos sedimentos e suas águas intersticiais. Os resultados indicam que houve, nos microcosmos-tratamento, um aumento do potencial redox do meio, um quase total abatimento do SVA dos sedimentos e uma redução do fósforo disponível nas águas em um nível bastante expressivo. As espectroscopias na região do UV-vis indicaram alterações nos perfis espectrais entre os microcosmos-controle e tratamento, sem, no entanto, identificar uma tendência uniforme ao longo dos períodos do experimento. A análise elementar foi concordante com os resultados de SVA e de espécies nitrogenadas nas águas intersticiais, pois mostram uma redução do teor de enxofre e de nitrogênio.
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Metody kvalitativní a kvantitativní analýzy PHA v buňkách cyanobakterií / Analytical methods for qualitative and quantitative determination of PHA in cyanobacteriaČernayová, Diana January 2020 (has links)
The diploma thesis is confused to verify the applicability of selected physicochemical and spectroscopic methods for characterization of cyanobacteria, with special emphasis on possibilities of qualitative and quantitative analysis of polyhydroxyalkanoates (specifically polyhydroxybutyrate (PHB)) accumulated in cyanobacterial cells. The sample basis of the work was formed by cultures of cyanobacterial strains of Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. The cultures were were cultivated in several ways to cover the widest possible range of physiological conditions and PHB contents, in particular using an autotrophic way of cultivation on shakers and multicultural culture method in a basic culture medium,and in media enriched with 2% salt (NaCl ) as well as mixotrophic culture media with different types of the carbon substrate. After few weeks of cultivation, cyanobacterial cultures were obtained and complexly analyzed by following techniques- cell suspensions were analyzed by flow cytometry and UV-VIS spectrometry (transmission and diffusion transmission mode), dry cell biomass was characterised by gas chromatography to obtain a exact amount of PHB, and then FT-IR spectrometry and thermogravimetric analysis. The work aimed to assess whether any of these methods can be a quick and affordable alternative to the determination of PHB content to the most commonly used method of gas chromatography, but also to assess what additional information about the physiological state of cyanobacterial cells can provide test methods. The highest correlation on PHB content was determined for the parameters determined by infrared spectroscopy, in which specific peaks from the characteristic wavelengths for polyhydroxybutyrate were important. Weak correlations on PHB content were achieved in thermogravimetric analysis and cytometry, using the hydrophobic fluorescent probe BODIPY 439/503, which bound to lipophilic parts of cells. In addition to the determination of PHB, it was possible to determine pigments present in cyanobacteria (such as chlorophyll, phycocyanin and carotenoids) by flow cytometry and UV-VIS diffusion transmission spectrometry. In the end, results from all used techniques were compared by PCA analysis to determine the similarity of all analyzed samples.
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Solubilizační schopnosti polysacharidů / Solubilizattion properties of polysaccharidesLenartová, Radka January 2008 (has links)
In this diploma thesis were studied solubilization properties of polysaccharides by using hydrophobic solutes (Sudan Orange G, Sudan Red G, (±)-alpha-Tocopherol, Pyrene, Perylene, Nile red), which were represented by alkyl derivates of hyaluronan. At first, a behaviour of individual hydrophobic solutes was investigated in variously polar solvents (Methanol, 1 Propanol, Chloroforme, Cyklohexane, n Heptane) and in the environment of varying ionic strength (water, 0.1 M and 0.4 M NaCl). Afterwards, solubilization properties of Sodium Dodecyl Sulfate model solubilizated the hydrophobic solutes into a core of micelles was examinate. We were interested in the solubilization capacity as the mol of solubilized molecules per mol micelles of surfactant corresponding with a state of micelles saturation. In the case of the solubilization of (±)-alpha-Tocopherol into the core of micelles, it was not possible to determine the solubilization capacity. So we changed the determination of universally solubilization power. The solubilization power is defined as mol of molecules solubilized per mol surfactant relative to the quantity solubilizate at the micelles saturation. Model system of Sodium Dodecyl Sulfate as a simple surfactant carrying a negative charge as the alkyl derivates of hyaluronan was selected bacause of its characteristics.The surfactant forms unimolar micelles and its critical micelle concentrations and aggregation numbers are tabelated for the investigated microenvironment. The main aim of the study was investigating of hydrophobic domains of alkyl derivates of hyaluronan as free places for incorporation hydropbobic solutes in the microenvironment of varying ionic strength. The critical aggregation concentrations were determined by the Pyrene 1:3 ratio method. For the research of micropolarity of alkyl derivates hyaluronan’s domains were selected two concentrations of derivates for the next research of solubilization experiments - the first concentration near the critical aggregation concentration and the second concentration above it. The effect of concentration of Pyrene on a core polarity of derivates was investigated. We discovered the influence of the concentration and the other we found a stationary area of the concentration. In the end we investigated the influence of preparation of solutions of derivates of hyaluronan on the core polarity by the concentration of pyrene which corresponds to the stationary area. The study of solubilization properties of alkyl derivates of hyaluronan is not a simple case as we assumed. When we measured spectra of the absorbance, higher concentration of derivates of hyaluronan belittle absorbance of solubilizates. At the experiment of solubilization with Sudan Red G we found out that Sudan Red G is not able to solubilizate into the hydrophobic core of micelles of hyaluronan’s derivates because of lipophilic or steric effects. We had to change Perylene as a new solubilizate. From the measured emission spectra we found saturation micelles. We can express the solubilization power of hyaluronan’s derivates for the concentration of Perylene. The main aim of the diploma thesis was to determine optimal way of the preparation of hyaluronan’s derivates solutions with required degree of solubilization.
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Analytická studie redukce rhenistanu amonného vybranými redukčními činidly / Analytical study of reduction of ammonium perrhenate using selected reduction agentsLišková, Jana January 2018 (has links)
This thesis is focused on analysis of reduction of ammonium perrhenate using less frequent reducing agents. Ascorbic acid and acetylhydrazine were chosen as less common reducing agents. Reductions were observed leveraging capillary zone electrophoresis and UV/Vis spectroscopy. Measurements were carried out both in acidic and in basic environments. Reduction of peak area of ammonium perrhenate was observed using capillary zone electrophoresis after addition of reducing agent. Rate of absorption of ammonium perrhenate was monitored with UV/Vis spectroscopy. Furthermore, the possibility of shifting the equilibrium of the reaction to the benefit of the reduction products was monitored by the addition of the complexing agent. Py- rogallol (benzene-1,2,3-triol) was used as the ligand and acetylhydrazine was selected as the reducing agent. The reduction was monitored by capillary zone electrophoresis. The identity of the resulting complex was confirmed with ESI/MS. Keywords rhenium, rhenium complexes, ascorbic acid, acetylhydrazine, capillary zone electrophore- sis, UV/Vis spectroscopy, mass spectrometry
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Srovnávací studie interakcí tenzidů s hyauronanem a jinými polyelektrolyty. / Comparative study of interaction between surfactant and hyaluronan and different polyelectrolytes.Stiborský, Filip January 2012 (has links)
In this diploma thesis, the interactions between polyelectrolyte and surfactant at low and also at high concentration were studied. There was used pyrene as fluorescent probe during the fluorescence spectroscopy measurement, a cationic surfactant cetyltrimethylammonium bromide and as a main polyelectolyte has been chosen sodium polystyrene sulfonate at 1 MDa molecular size. In the medium containing 0.15 M NaCl we could observed a creation of the complexes – precipitates in the surrounding of CMC concentration and behind of this concentration. In the mixtures containing sodium polystyrene sulfonate and hyaluronan together, there was stronger tend to keep aggregation properties of sodium polystyrene sulfonate during difference concentration ratios. Beyond CMC concentration, hyaluronan starts to influence the aggregation properties of the system as well.
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Fluorosolvatochromism of furanyl- and thiophenyl-substituted acetophenonesFriebe, Nadine, Schreiter, Katja, Kübel, Joachim, Dietzek, Benjamin, Moszner, Norbert, Burtscher, Peter, Oehlke, Alexander, Spange, Stefan 15 February 2016 (has links) (PDF)
A series of para-substituted acetophenones bearing a furanyl or a thiophenyl moiety show a large Stokes-shift, which is a function of various solvent properties. Photophysical properties such as emission lifetime of the compounds have been determined using time-correlated-single photon counting to secure the intrinsic fluorescence behaviour. The solvent dependent position of the UV/Vis emission band [small nu, Greek, tilde]max,em of the compounds has been measured in 26 various solvents. The influence of the solvent on [small nu, Greek, tilde]max,em is of very complex nature and mathematically analysed by multiple square linear solvation energy (LSE)-correlation analysis using Catalán's four-solvent parameter set. Solvent acidity has a strong influence on the bathochromic shift of 2,5-disubstituted furan derivatives compared to the non-5-substituted furan and thiophene derivatives, which show a contrary behaviour. Therefore, the 5-cyanofuranyl-substituted acetophenone derivative is useful as a probe for measuring environmental properties by fluorescence spectroscopy. / Dieser Beitrag ist aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
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Komplexy 2,6-bis[(N-methylpiperazin-1-yl)methyl]-4-formyl fenolu / Complexes of 2,6-bis[(N-methylpiperazine-1-yl)methyl]-4-formyl phenolMarečková, Vendula January 2012 (has links)
Thirteen new dinuclear complexes of ligand 2,6-bis[(N-methylpiperazine-1- yl)methyl]-4-formyl phenol (L1) were prepared. Copper(II) and palladium(II) salts were used for the syntheses. Following compounds were prepared: [Cu2L1(CH3COO)2]ClO4, [Cu2L1(CF3COO)2(ClO4)][Cu2L1(CF3COO)2]ClO4, [Pd2L1(CH3COO)3], [Pd2L1(CF3COO)3], [Pd2L1(Cl)4]. These ones should act as anion receptors. The acetates were exchanged for phenylphosphinates in the case of the complex cation [Cu2L1(CH3COO)2]+ and the structure of [Cu2L1(phPO2H)2]ClO4 was obtained. Geometry of coordination shell of Cu2+ - ions in prepared complexes is tetragonal pyramid. The UV-VIS spectroscopy was used for study of interactions in system of the ligand - metal - anion. Interactions between the ligand and metals in oxidation state 2 were observed as well as interactions between the complex [Cu2L1(CH3COO)2]ClO4 and sodium salts.
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Spektroskopische Charakterisierung der grün-absorbierenden Kanalrhodopsin-Chimäre ReaChRKrause, Benjamin Sören 06 September 2018 (has links)
Kanalrhodopsine (ChRs) sind lichtgesteuerte Ionenkanäle, welche nach Absorption eines Photons durch den Retinal-Cofaktor einen passiven Ionentransport über die Zellmembran katalysieren. Im Zuge von optogenetischen Anwendungen wird diese Reaktion für die Beeinflussung der Ionenhomöostase von verschiedenen Zelltypen und Geweben ausgenutzt. Zu Beginn dieser Arbeit wurden lichtinduzierte Strukturänderungen und Protontransferschritte in einem breiten Zeitbereich (Nanosekunden bis Minuten) in dem grün-absorbierenden ChR ReaChR mithilfe von stationärer und transienter UV-vis- und Fourier-Transform-Infrarot-Spektroskopie (FTIR) untersucht. Auf Basis der experimentellen Daten wurde ein komplexes Photozyklus-Modell konzipiert.
Anschließend wurde die IR-aktive, nichtkanonische Aminosäure p-Azido-L-phenylalanin (azF) mittels Stopp-Codon-Suppression ortsspezifisch an mehreren Positionen innerhalb der vermuteten ionenleitenden Kanalpore in ReaChR inkorporiert und mit FTIR untersucht. azF ist sensitiv gegenüber Polaritätsänderungen und absorbiert in einem hochfrequenten Bereich (~2100 cm-1). Aufgrund der großen spektralen Separation zu endogenen Proteinschwingungen (< 1800 cm-1) können globale Konformations- und lokale Hydratisierungsänderungen simultan detektiert werden. Die erhobenen Daten leisten einen wichtigen Beitrag zum Verständnis der Bildung einer temporären Wasserpore in ChRs und demonstrieren zum ersten Mal den erfolgreichen in-vivo-Einbau einer artifiziellen Aminosäure in mikrobielle Rhodopsine und dessen schwingungsspektroskopische Analyse. Die Methode bietet aufgrund ihrer hohen Ortsauflösung ein großes Potential für die Studie von Mikroumgebungen innerhalb komplexer Proteinensemble. / Channelrhodopsins (ChRs) are light-gated ion channels. Upon absorption of a photon, the retinal chromophore isomerizes and drives conformational changes within the protein, which lead to a passive ion transport across the cell membrane. This capability is used for optogenetic applications to manipulate ionic homeostasis of different cell types and entire organisms. Within the work, light-induced structural changes and proton transfer steps were studied in the green-absorbing ChR ReaChR in great detail by steady-state and transient UV-vis and Fourier transform infrared spectroscopy (FTIR). The data were merged into a complex photocycle model.
Next, the IR-active, unnatural amino acid p-azido-L-phenylalanine (azF) was site-specifically introduced at several sites of the putative ion pore of ReaChR by stop codon suppression. azF is sensitive to polarity changes and absorbs in a clear spectral window lacking endogenous protein vibrations. Thus, FTIR measurements of labeled mutants report for global conformational changes (< 1800 cm-1) and local hydration changes (~2100 cm-1) simultaneously. The presented findings reveal crucial insights regarding formation of a transient water pore in ChRs and demonstrate the first report of the successful in-vivo incorporation of an artificial amino acid into a microbial rhodopsin and its subsequent spectroscopic investigation. Additionally, the so far unprecedented spatial resolution renders this methodology superior over conventional FTIR methods to study microenvironments within complex protein ensembles.
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Self-Organization of β-Peptide Nucleic Acid Helices for Membrane ScaffoldingHöger, Geralin 14 February 2019 (has links)
No description available.
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