The subject of the doctoral thesis is the study of moisture resistance of macro-defect-free (MDF) composites based on calcium-aluminate cement and polyvinylalcohol used as polymer. MDF cements are perspective materials providing unique properties relative to traditional cement pastes with great potential to incoming constructional utilization. But it is known that MDF cements exhibit sensitivity to water, with swelling and reduction of strength. Therefore the work is aimed in monitoring of an organic polymer effect on the moisture resistance of MDF materials and phase changes. The effects of three different organotitanate cross-linking agents on the properties were investigated. The organotitanate-modified MDF cement samples exhibited improved moisture resistance as compared to the standard samples when exposed to 100 % relative humidity or water at laboratory conditions. The influence of composition at different curing on the properties of model MDF cement samples was observed. The characterization of MDF samples was based especially on test of flexural strength, XRD, DTA/TGA and EGA, EDS and scanning electron microscopy analyses.

Some proteins require presence of their specific ligand, cofactor or prosthetic group for their activity. Binding of this specific molecule can cause conformational changes which permit to perform their function. In some occasions the identification of conformational changes could be really challenging task. In this thesis we describe the novel approach for monitoring structural changes in proteins using chemical cross-linking and high resolution mass spectrometry and its application on model calmodulin system. It is demonstrated that analysis using isotope-labelled cross-linking agents enabled us to get insight into the structural rearrangement caused by presence or absence of the protein ligand. However, it is shown that the method has potential drawback due to limited enzymatic proteolysis. The novel approach that also makes it possible to quantify the changes in protein structure was used together with other methods for characterization of the neutral trehalase Nth1 in complex with Bmh1 protein (yeast isoform of protein 14-3-3). The results revealed that Bmh1 induce structural rearrangement of Nth1 molecule with changes within the EF- hand like motif which is essential for the activation process.

in English β-N-acetylhexosaminidase (EC 3.2.1.52) belongs to exoglycosidase, and is one of the most abundant enzymes found in organisms from bacteria to human. The fungal β-N-acetylhexosaminidase from Aspergillus oryzae is composed of propeptide and catalytic domain. The propeptide is noncovalently associated with the catalytic domain of the enzyme. Propeptide is essential for the enzyme activity. While the structure of the catalytic domain was desidned by homology modeling, the structure of the propeptide has not been resolved yet. In this study, the position where the propeptide is associated with the catalytic domain, was uncovered. Presented work consists of two parts. First part deals with optimization of production conditions, purification and crystallization of β-N-acetylhexosaminidase from the filamentous fungi Aspergillus oryzae. Second part is devoted to the study of interaction between propeptide and catalytic domain, which was characterized by chemical cross-linking and high-resolution mass spectrometry. It was found that the structural changes of the catalytic domain depend on the presence of the propeptide molecule. Moreover the region of propeptide-catalytic domain interaction was revealed.

No description available.

No description available.

Background and objectives: TISSIUM light-activated adhesive was investigated as an alternative to tissue-penetrating products to fix meshes in intraperitoneal laparoscopic ventral hernia repair. The objective of this study was to ensure efficient polymer light activation through commercial meshes and to assess the acute and chronic fixation strength of the light-activated adhesive in a porcine model in comparison to commercial fixation products. Methods: A spectroscopic analysis was conducted on the light-activated adhesive through three different meshes (1, 2, and 3) to quantify the acrylate conversion associated with the level of polymer cross-linking. Two setups were implemented: a static (light source fixed over a drop of polymer) and a dynamic (light source rotated around a pattern of polymer to mimic the surgical procedure). Hernia defects were created in porcine models and repaired either using the light-activated adhesive or a commercial product (A, B, C, and D) to fix a mesh. For each tested condition, the acute and chronic (3 months) fixation strength performances were assessed using burst ball and t-peel mechanical tests. Results: The light activation proved to be effective (more than 90% of the acrylates converted) in static in 7 seconds through the three meshes and in dynamic between 3 min and 5 min 32 sdepending on the considered mesh. In a burst ball test, the light-activated adhesive reached between 42 and 84% of the commercial products’ acute performance with the three meshes (between 75,9 and 95,9 N) and reached 88% of the commercial product A’s chronic performance with mesh 1 (610,1 N). A t-peel test demonstrated similar strength of ingrowth for the repairs using the light-activated adhesive or the commercial product A at the 3-month timepoint with mesh 1 (2,55 and 2,37 N/cm respectively). Conclusions: Data suggest the light-activated adhesive has the potential to be used in intraperitoneal laparoscopic ventral hernia repair. In a reasonable time, the adhesive is efficiently light-activated through commercial meshes. The light-activated adhesive’s performances to fix commercial meshes, both acute and chronic, are similar to commercial products, but with a strong advantage of not being tissue penetrating.

The goal of this research was to utilize the morphological control inherently imparted by the electrospinning process to improve the active layer performance in energy conversion devices as well as to better understand the relationship between morphology and performance in energy storage devices. Discrete control of the active layer morphology can promote exciton dissociation in organic photovoltaic cells (OPVs), whereas developing efficient ion diffusion pathways and beneficial polymer-ion interaction in polymer-gel electrolytes is demonstrated to result in enhanced battery performance. We demonstrate the ability to develop unique morphologies in Poly(3-hexafluoro propylene) (P3HT) films with energy storage applications using various electrospinning techniques. Electrospinning in a solvent-saturated atmosphere allows for the design of ribbon architectures with polymer domains on the order of 5-10 um. These ribbon structures form what appear to be bi-continuous films, which could then be filled with an acceptor / fullerene type material to create a bulk heterojucton for OPV devices. Dropping chloroform onto the electrospinning needle during the spinning process results in P3HT fibers with porous surfaces. These fibers have diameters of ~ 2 um. Using a coaxial needle to electrospin a P3HT solution in the core, and a CHCl3 sheath solution created hybrid ribbon-fiber structures. These structures have even smaller domain sizes than the ribbons created using a solvent saturated atmosphere. Cospinning P3HT with sacrificial polymers results in P3HT fiber morphologies upon removal of the sacrificial template polymer. Additionally, introducing P3HT into an established fiber matrix results in fibrous P3HT architectures after the template fibers are removed. Developing hybrid polymer-gel electrolytes using crosslinked PEO electrospun fibers results in membranes with high affinity for liquid electrolyte components. These electrospun PEO fiber mats exhibit excellent ionic conductivities at room temperature (12 mS/cm) exceeding an electrospun PVDF control. Furthermore, the PEO fiber mats can absorb nearly three times as much liquid electrolyte as the PVDF control. PEO has been show to interact with lithium salts to aid in dissociation and diffusion during battery cycling. Although the ionic conductivity data suggest PEO to be a superior electrolyte, pulsed-field-gradient NMR shows that lithium diffusion is faster in PVDF samples. From coin cell discharge experiments, PEO is believed to interact strongly with Li+ ions, inhibiting them from diffusing rapidly during fast charge/discharge rates. However, PEO/PETA fiber electrolytes show nearly 100% theoretical capacity discharge at C/100 and a capacity retention of ~ 35% at a C/5 discharge rate in contrast to a glass fiber separator which shows only a capacity that is approximately 85% of the theoretical value. The unique mechanical properties of PEO/PETA electrospun mats could lead to interesting artificial skin and wound healing applications. Upon crosslinking at elevated temperatures (~40 degrees C), the fiber mats exhibit improved tensile strength and much higher ultimate stress at break. The porous nature of the materials lend to easy oxygen diffusion for wound healing, and the hydrophilicity promotes continued adhesion to existing tissue making these mats possible adhesive-less bandages. / Ph. D.

DCL-1 (CD302) is a type I transmembrane C-type lectin receptor, which is expressed on monocytes, macrophages, granulocytes and dendritic cells. However, its extracellular domain lacks the amino acids motives essential for carbohydrate binding in the presence of calcium ions, suggesting that it does not have the classic binding capacity found in other C-type lectin receptors such as the mannose receptor. No exogenous or endogenous ligands have been identified yet, though. Due to internal colocalization with F-actin we can assume, that this unconventional lectin receptor plays a role not only in endocytosis and phagocytosis but also in the cell adhesion and migration. The receptor DCL-1 was first identified as a genetic fusion partner of human DEC-205 multilectin receptor in Hodgkin's lymphoma cell lines. The experimental part of this thesis deals with the characterization of disulfide bonds and data acquisition for validation of DCL-1 crystal structure. First the production and refolding conditions were optimized to obtain the highest amount of DCL-1 protein, precisely its extracellular domain. These optimal conditions were used to prepare the protein for in-gel digestion using specific endopeptidases in the presence of cystamine followed by LC-MS analysis. DCL-1 disulfide bonds were determined by comparing...

A comprehensive understanding of physiological role of proteins requires knowledge of their three-dimensional structure, dynamics and protein-protein interactions. Chemical cross-linking in combination with mass spectrometry represents an alternative approach to standard methods for protein structure elucidation (X-ray crystalography, NMR spectroscopy) and enables characterization of interaction interface within protein complexes in their native states. The presented thesis is mainly focused on novel cross-linking methodology based on the in vivo incorporation of methionine analog with photo-reactive functional group (photo-Met) into the sequence of studied protein (so called protein photo-nanoprobe). Interaction between two molecules of 14-3-3zeta protein was used as a model to test and optimize the protein photo-nanoprobe production. The findings confirmed usefulness of this approach for mapping the protein-protein interactions. The photo-initiated cross-linking was used to detect the heterooligomeric membrane structures of cytochromes P450 2B4 and b5 and the molar ratio of cytochromes within individual complexes was assessed. The chemical cross-linking in combination with mass spectrometry was employed to characterize the interaction of their catalytic domains and two mutual orientations of...

As espumas de polietileno são amplamente utilizadas. Suas aplicações vão de aparelhos domésticos a artefatos médicos, incluindo aplicações nas indústrias de construção civil e automobilística. As propriedades das espumas dependem da densidade e da sua estrutura celular, em outras palavras, da quantidade de células abertas e fechadas, da distribuição e do tamanho das células. Os métodos de produção de espumas de polietileno reticulado são classificados em dois tipos, de acordo com o método de reticulação. Um dos métodos é a reticulação química, que utiliza peróxidos como agente reticulante, O outro método é a reticulação por irradiação, utilizando feixe de elétrons. As espumas obtidas a partir do polietileno reticulado pelo processo de irradiação apresentam superfície lisa e homogênea, e são formadas basicamente por células fechadas. O objetivo deste trabalho foi utilizar a radiação ionizante para a formação de ligações cruzadas entre as moléculas de polietileno de baixa densidade (LDPE), que permitiu a obtenção das espumas. As suas propriedades mecânicas, térmicas e morfológicas foram avaliadas. As amostras de polietileno com densidade de 0,946 g/cm3, contendo 5% de azodicarbonamida (ADCA), agente expansor, foram irradiadas com feixe de elétrons de alta energia com doses de radiação de 10, 20, 30, 40, 50, 60, 80 e 100 kGy. Após a irradiação, as amostras de LDPE foram colocadas em um forno para expansão térmica e formação das espumas. Foi determinado o grau de reticulação das espumas. Algumas amostras também foram envelhecidas termicamente. O desempenho mecânico das amostras de espumas foi avaliado por meio de ensaios de tração e deformação, dureza, deformação permanente por compressão e resiliência. Também foram realizados ensaios de termogravimetria (TGA) e de microscopia eletrônica de varredura (MEV). Os resultados mostraram que no intervalo de doses de radiação estudadas, a resistência à tração aumenta com o aumento do grau de reticulação. Nos ensaios de resistência à compressão, a partir da dose de radiação de 40 kGy, observou-se um decréscimo nos valores obtidos. Já a resiliência sofreu pouca aheração. De acordo com a análise morfológica pode-se concluir que a dose de radiação de 30 kGy foi a que promoveu a formação de espumas com estrutura celular fechada e mais homogênea. / The polyethylene foams are widely used. Their main applications are used for both home appliances to medical equipments. Beside that, they have applications in building and automotive industries. The foam properties depend on the density and its cellular structure, that is, the amount of open and closed cells, of the distribution and size of them. The methods of the crosslinking polyethylene foam production are classified in two types, according to the crosslinking method. One method is based on the chemical crosslinking, which utilizes peroxide as crosslinking agent. In the other method, the crosslinking is induced by electron beam radiation. The foams obtained from the crosslinking polyethylene by irradiation process presented a smooth and the homogeneous surface, and are formed basically by closed cells. The aim of this study was to apply the ionizing radiation from electron beam to crosslink low density polyethylene (LDPE), to obtain foams. Their morphological, thermal and mechanical properties were studied to evaluate the obtained samples. The samples of low density polyethylene (0,946 g/cm3), containing 5% of azodicarbonamide (ADCA), as expander agent, were irradiated with electron beam with doses of 10, 20, 30, 40, 50, 60, 80 and 100 kGy. After the irradiation, these LDPE samples were put into an oven to obtain the foams. It was determined the crosslinking degree of the foams. Some samples were also thermically aged. The mechanical performance of the foams samples was evaluated by means of the tensile strenght, compression, hardness, permanent deformation by compression and resilience. It was also carried out thermogravimetry and scanning electron microscopy (SEM). The results have shown that, in the interval of radiation doses studied, that the tensile strength increases with the increase of the crosslinking degree. The compression resistance results obtained from with samples with to radiation dose of 40 kGy showed significant decreasing. The resilience measurements have shown only little variations. According to the morphological analysis it can be concluded that foams with radiation dose of 30 kGy have closed cellular structure more homogeneous and smooth morphology.