This master’s thesis is focused on the network architecture of IMS (Internet Protocol Multimedia Subsystem) and development an application for IMS. IMS architecture is the convergence of fixed and mobile networks. IMS enables rapid deployment of new multimedia services. In the first chapter is showed and described architecture IMS in four layers and block model of architecture IMS. Next chapter is focused to protocols, which are used in IMS. One of the most important protocols in network IMS is SIP. This protocol is analyzed in details. For development IMS platform application was used Ericsson SDS (Service Development Studio) in version 4.1. In SDS were created applications type of client and servlet. In this master’s thesis is also used development environment NetBeans. In NetBeans was created application for management of captured images. In the same development environment was developed application that shows the contents of a database. Last chapter is devoted to analyze individual parts of communication between client and servlet, and when client uses http to download image file.

This diploma thesis deals with the analysis of low molecular weight proteins in barley during malting by SDS-PAGE method. Attention was paid to PR proteins, specifically LTP proteins and thionins, considered to be connected with gushing effect. Barley samples, malt intermediates and malt samples taken in 10 consecutive days to cover the whole malting process were used for the experiment. In total, 5 samplings were used for the analysis. Proteins extracted from the samples were separated by SDS-PAGE using a Tris-tricine buffer system. The protein lines of LTP proteins and thionins were identified in the resulting gels. The relative optical density values of the selected protein bands were obtained to assess the effect of malting on these proteins. A similar pattern of change in the content of mentioned low molecular weight proteins during the malting process was observed. This was confirmed by finding a statistically significant positive correlation between the relative optical density values of LTP proteins and thionins. Furthermore, the relationship between the low molecular weight protein content and the gushing potential and the microbiological contamination of the samples was investigated, but was not confirmed.

The MS-818 compound is used in the proliferation process of neuronal cells and many biological activities that accompany this process such as astrocyte differentiation, inhibition of neuronal apoptosis, and fraction repairs. We do know the effects of this compound, but the mechanism of action remained uncertain until now. To determine the pathway of this compound, NT2 cells were cultured and lysed to isolate the proteins. Affinity Chromatography was performed in order to immobilize the MS-818 compound to a Hi-Trap NHS column. The NT2 protein sample was injected through the column and eluted with a MS-818 concentrated, high salt content elution buffer. SDS-PAGE was then performed to isolate the proteins that bound to MS-818. The gel was visualized using Coomassie Blue. The results indicate that there are two proteins associated in the mechanism of this compound. A standard protein marker ranging from 10 kDa to 250 kDa was used to compare the bands. The findings indicate that one of the protein bands is slightly less than 250kDa and the other is between 50-75 kDa. When the proteins are confirmed by mass spectrometry sequencing, this will help to promote this compound as a drug candidate.

Experiments were conducted to test for ricin content in tissue samples from four castor cultivars, developing castor seed, germinating castor seedlings, and chemically and heat treated seed meal. Ricin content of each sample was examined via Western blotting with ricin A-chain specific antibodies. Results indicate that ricin is present solely within castor endosperm and is not present any other tissues. Samples from developing seed and germinating seedlings indicate ricin production begins around day 28 post pollination, and ricin is absent from the seedling 6 days after the onset of radicle emergence. This would seem to indicate that the purpose of ricin is to protect the seed and not the entire plant. Ricin content of seed meal treated separately with heat and chemicals was tested. It was found that hot-pressing of the seed was sufficient to denature ricin in the seed meal.

No description available.

Reproduction is an essential feature of all animals and a fundamental step to produce new generations. Study of sperm proteins is crucial for understanding of the sperm-egg recognition. We searched out sperm surface proteins involving in the zona pellucida (ZP) binding and studied whether these proteins are preserved throughout mammalian species. Indirect immunofluorescent technique was used to test a panel of monoclonal antibodies prepared against boar sperm surface proteins on spermatozoa of bull and mice. We found a cross-reactivity of some antibodies against boar sperm with bull ejaculated and mouse epididymal spermatozoa. Further, we isolated sperm proteins from different mammalian species, such as pig, bull, dog, cat, mouse and human. Proteins were separated by SDS- electrophoresis and protein/glycoprotein profiles from epididymal, ejaculated and in vitro capacitated sperm were compared. The interaction of sperm with ZP was studied on electrophoretically-separated sperm surface proteins from pig and bull with biotin-labeled ZP glycoproteins. Antibodies, which reacted with boar sperm surface proteins with ZP- binding activity, therefore could be potential egg-binding receptors, were used for monitoring of the sperm protein origin in reproductive fluids and tissues. (In Czech) Keywords: sperm...

A Terapia Fotodinâmica (TFD) é um método de tratamento em que um composto fotossensibilizador (FS) é introduzido no organismo do paciente e, posteriormente, a região de tratamento é irradiada com luz. A combinação de oxigênio, luz e FS produz espécies que geram um efeito curativo. Os FS utilizados atualmente, embora eficazes, apresentam desvantagens. Neste trabalho, foram estudados os corantes ciânicos com dois cromóforos ou biscianinas (BCD) que apresentam características favoráveis na aplicação como FS na TFD. O objetivo deste trabalho foi analisar os efeitos da concentração, da composição do ambiente e da própria estrutura de três BCDs, com ângulos entre os seus cromóforos de 180°, 150° e 90°, nas características das suas fototransformações em tampão e na interação com micelas de SDS e DNA. A fototransformação pode causar a perda da fotoatividade do FS e gerar fotoprodutos (FPs) que tanto podem ser tóxicos como fotoativos. Os processos de tratamento na TFD e os de fototransformação são paralelos. Isso permite utilizar o monitoramento do processo de fototransformação para determinar o grau do processo de tratamento em tempo real. Por esse motivo, também foi estudada a fototransformação dos FPs formados durante a irradiação. A análise da fototransformação foi realizada utilizando técnicas espectroscópicas de absorção óptica e de fluorescência estática. Mudanças nos espectros de absorção óptica dos corantes indicaram a formação de agregados. A agregação diminui a velocidade de fototransformação devido ao aumento da probabilidade de dissipação da energia de excitação por processos não radiativos. Não houve formação de agregados na irradiação dos FPs, pois estes possuem menor tendência de agregação em comparação com o próprio corante. Semelhanças nos espectros dos FPs dos três corantes indicaram que eles possuem a mesma estrutura do cromóforo. Na ausência do oxigênio a velocidade de fototransformação diminui. Este fato juntamente com a relação entre os tempos da fotólise e os rendimentos quânticos do estado tripleto dos BCDs sugeriram a participação do oxigênio singleto na fototransformação. Na presença de sistemas biomiméticos em altas concentrações, os corantes estão em suas formas monoméricas. Nestas interações, os corantes também sofreram fototransformação, com velocidades diferentes daquelas em tampão, com formação de fotoprodutos. Na fototransformação dos corantes na presença de micelas de SDS, sugerimos que as moléculas dos corantes ficam mais protegidas do contato do oxigênio. Por outro lado, na interação com o DNA, as moléculas dos corantes ligadas na superfície do DNA se tornam mais suscetíveis ao ataque do oxigênio, fazendo com que a fototransformação seja mais rápida. / Photodynamic therapy (PDT) is a treatment that a photosensitizer compound (PS) is inserted into the patient\'s body and, subsequently, the treatment area is irradiated with light. The combination of oxygen, light and FS produce species that generate a curative effect. The FS currently used, although can be effective, have disadvantages. In this study, the cyanine dyes with two chromophores (BCD) or biscyanines that have favorable characteristics in the application as FS in PDT were studied. The aim of this study was to analyze the effects of concentration, environmental composition and the structure of three BCDs, with angles between chromophores 180, 150 and 90 degrees, in the characteristics of phototransformation of the dyes molecules in phosphate buffer and in the interaction with SDS micelles and DNA. The phototransformation can cause the loss of the FS photoactivity and generate photoproducts (PPs) that are both toxic as photoactive. The treatment processes in PDT and phototransformation are parallel. It allows the use of phototransformation monitoring process to determine the degree of realtime treatment process. For this reason, it was also studied the phototransformation of PPs formed during irradiation. The phototransformation analysis was performed using optical absorption and fluorescence spectroscopies. Changes in the optical absorption spectra of the dyes indicated the formation of aggregates. The aggregation reduces phototransformation rate due to the increased probability of dissipation of the excitation energy by non-radiative processes. There was no aggregate formation on irradiation of PPs because they have less tendency to aggregate compared with the dye. Similarities in the spectra of the three PPs indicated that they have the same structure of the chromophore. In the absence of oxygen phototransformation rate decreases. This fact together with the relationship between photolysis time and the quantum yields of the triplet state of the BCDs suggested the involvement of singlet oxygen in their phototransformation. The dyes were in their monomeric form at high SDS and DNA concentrations. In these interactions, the dyes also were phototransformed, with different rates compared with phosphate buffer, with formation of photoproducts. In the phototransformation of dyes in the presence of SDS micelles, we suggest that the dye molecules were more protected from oxygen contact. On the other hand, in the interaction of dyes and DNA molecules, the dye molecules bound on DNA surface become more available to oxygen attack, and the phototransformation becomes faster.

Made available in DSpace on 2017-07-10T17:37:45Z (GMT). No. of bitstreams: 1 Patricia_Paro.pdf: 402899 bytes, checksum: cb9972fdc35b447bcb1020e13a07782b (MD5) Previous issue date: 2011-04-08 / Fundação Araucária / The wheat is a species of great importance in human nutrition, wheat grain is extracted from wheat flour used in the preparation of various food products. The rheological properties of wheat flour is a feature that determines the fate of the flour, being of great importance to this assessment. In breeding programs for wheat determining the quality of flour is essential for the development of cultivars, and it is necessary that the reading of the quality parameters are carried out early, with the use of small amounts of sample, reliable and the level of genotype. The development of this work aimed at the validation of molecular markers for selection of genotypes from high molecular weight glutenin, given their great influence on baking quality of wheat flour and check the correlation between parameters of quality of flour and the high molecular weight glutenin. Analyses were performed in the laboratory of biotechnology COODETEC using 77 samples of wheat (cultivars and lines) from program for wheat breeding of the institution, with data quality technology of wheat flour has characterized. The set of primers called Glu1-DX2-DX5 was used to select individuals that contained subunits Glu1-DX5 + Dy10 by PCR, nevertheless was necessary to make adjustments so that the marker amplified a fragment corresponding to allele Glu1-DX5. To check the reliability of molecular marker, all samples were characterized by electrophoresis of proteins by SDS-PAGE method and scores as a function of the subunits of glutenin high molecular weight they have. The results of the analysis of protein and molecular differ, indicating that the use of molecular markers should be used with any control in the PCR reaction. From the data collected and the quality of flour, the results were submitted to Pearson correlation analysis. It appears that the score of the protein is positively correlated with W, P and P / L, indicating that the composition of glutenin high molecular weight have significant influence on quality of wheat flour. Alleles Glu1-DX5 and the subunits of the A genome are positively correlated with protein scores and consequently with the wheat breadmaking. The allelic variants of the A genome were positively correlated with W and P. From the results it is concluded that the selection of genotypes with superior quality of flour can be made taking into account the high molecular weight glutenin encoded by the A and D genomes of wheat / O trigo é uma espécie de grande importância na alimentação humana, de seus grãos é extraída a farinha de trigo utilizada no preparo de diversos produtos alimentícios. As propriedades reológicas da farinha de trigo são características que determinam o destino final da farinha, sendo de grande importância a sua avaliação. Em programas de melhoramento genético do trigo a determinação da qualidade de farinha é indispensável para o desenvolvimento de cultivares, e é necessário que a leitura dos parâmetros de qualidade sejam realizados precocemente, com a utilização de pouca quantidade de amostra, confiáveis e a nível de genótipo. Este trabalho teve por objetivo a validação de marcadores moleculares para a seleção de genótipos a partir de gluteninas de alta massa molecular, visto que apresentam grande influencia na qualidade de panificação da farinha de trigo e verificar a correlação existente entre parâmetros de qualidade de farinha e as gluteninas de alta massa molecular. As análises foram realizadas no laboratório de biotecnologia da COODETEC utilizando 77 amostras de trigo (linhagens e cultivares) provenientes do programa de melhoramento genético de trigo da instituição, com dados de qualidade tecnológica da farinha de trigo já caracterizados. O conjunto de primers denominados Glu1-Dx5-Dx2 foi utilizado para selecionar indivíduos que continham o conjunto de subunidades Glu1-Dx5+Dy10 através da técnica de PCR, porém foram necessário ajustes para que o marcador amplificasse o fragmento correspondente ao alelo Glu1-Dx5. Para verificar a confiabilidade do marcador molecular todas as amostras foram caracterizadas através de eletroforese de proteínas pelo método SDS-PAGE, e escoreadas em função das subunidades de gluteninas de alta massa molecular que apresentam. Os resultados entre a análise de proteína e molecular apontam divergências, indicando que para a utilização do marcador molecular deve ser utilizada com algum controle na reação de PCR. A partir dos dados coletados e os de qualidade de farinha os resultados foram submetidos a análise de correlação de Pearson. Verifica-se que o escore de proteína é positivamente correlacionado com W, P e P/L, indicando que a composição de gluteninas de alta massa molecular apresentam influencia significativa na qualidade de farinha de trigo. Os alelos Glu1-Dx5 e as subunidades do genoma A estão positivamente correlacionados com escore de proteína e conseqüentemente com a qualidade e panificação do trigo. Os variantes alélicos do genoma A foram positivamente correlacionados com W e P. A partir dos resultados conclui-se que a seleção de genótipos de trigo com qualidade superior de farinha podem ser realizados levando em consideração as gluteninas de alta massa molecular codificadas pelos genomas A e D do trigo

<p>Obesity is a globally increasing disease particularly in developing countries and among children. It is mainly caused by intake of diets high in fat and the lack of physical activity. Obesity is a risk factor for diseases such as type II diabetes, high blood pressure, high cholesterol and certain cancers. Prediabetes is a condition where blood glucose levels are above normal but have not&nbsp / reached those of diabetes. It is difficult to diagnose, as there are no signs or symptoms. Some type II diabetes patients bear no symptoms at all and the disease is discovered late. Proteomics is a field that can provide opportunities for early diagnosis of diseases through biomarker discovery. The early diagnosis of diabetes can assist in the prevention and treatment of diabetes. Therefore there is a need for the early diagnosis of diabetes. Twenty Wistar rats were used. The rats were initially fed a CHOW diet, which is the standard balanced diet for rats, for 4 weeks. The rats were then divided into 2 groups of 10 where 1 group was fed CHOW and another was fed a high fat (HF) diet in order to induce obesity. The two groups were fed their respective diets for 18 weeks. Rats were weighed. Rats were placed in metabolic chambers and 24 hour urine samples were collected. Ketone levels were measured by Ketostix. Urine proteins were precipitated by acetone, quantified and separated on both the 1D SDS-PAGE and the 2D SDS-PAGE. Protein expression changes between CHOW and HF fed rats were determined and identified using MALDI-TOF mass spectrometry. Protein spots intensities increased and decreased between the CHOW and HF fed rats. Transducin (beta)-like 3 was identified as the only differentially expressed protein, which might serve as a potential biomarker for prediabetes.</p>

Oily wastewater produced from petroleum and petrochemical refining processes is one of the gravest environmental threats. Oil waste ending up in sewers and dumps each year is equal to 25 times the amount of crude oil spilled in the Exxon Valdez accident (1989). Oil/Water separation covers a broad spectrum of industrial process operations. There are many techniques employed depending on each situation. The byproduct of water recovery from oily wastewater is a sludge rich in oil, surfactants and particles (oily sludge). The oily sludge still contains significant amounts of waters, which need to be recovered prior to its disposal. The use of polyelectrolytes for the flocculation of the emulsified oil and its separation from the aqueous phase is usually one of the steps of the wastewater as well as oily sludge treatment process. The efficiency of polyelectrolytes as floculants is quite often evaluated via trial and error and the appropriate polymer is selected according to the case. Even in scientific investigations it is rather common to use industrial oily sludge samples. The industrial oily sludge is characterized and treated by polyelectrolytes. Nevertheless industrial oily sludge is quite complicated and variable to be approximated by a model. For the systematic study of polyelectrolytes efficiency a stable, realistic and well-defined oily sludge model is necessary. In the present work an oily sludge model was successfully developed and characterized. The model consists on water, oil (kerosene), surfactant (sodium dodecyl sulphate, SDS) and clay particles (Blauton). The emulsifying efficiency of the surfactant and the clay were studied independently. The interactions between the surfactant and the clay including adsorption of the former on the later and cation exchange reactions were investigated. The four components were finally combined to form a series of emulsions varying the relative amounts of the emulsifiers for the highest stability to be encountered. Having concluded on the composition of the oily sludge model the efficiency of various polyelectrolytes was evaluated. Commercial natural (chitosans), synthetic (PolyDADMACs) polyelectrolytes and oilbreaks as well as lab-scale semi-synthetic polymers (modified chitosans) were tested. The flocculation efficiency was determined based on the amount and quality of water that was recovered as well as on the floc stability, size and sedimentation speed. The recovered water was characterized according to the environmental protection agency (EPA). The analysis included measurements on: total organic carbon (TOC), chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total suspended solids (TSS) and pH. Selection of the appropriate flocculant also depends on the type of flocs formed in combination to the treatment following the flocculation. When filtration or centrifugation is used as a post-flocculation process the appropriate polymers are those that form large and porous flocs, as the case of the modified chitosans AG95 and AG97. Clarification devices on the other hand require dense flocs as these produced by the use of PolyDADMACs, commercial chitosans, oilbreaks and modified chitosans GA35, GA41 and AG79. Regarding the water quality, some of the polymers used that have low values of COD, TOC and BOD5, may not need the secondary treatment (biological) prior to discharge, such as P187K (PolyDADMAC) and GA41 (modified chitosan). The others require a biological treatment for the regulation limits to be reached. The pH values of modified chitosans (except of AG97), lab-scale PolyDADMACs and an oilbreak (OCAA) are all in range of the regulation limits. The applicability of biopolymers as flocculants for oil sludge dewatering is a relatively new field of investigation. As a consequence of the growing demand for environmentally friendly technologies as well as renewable resources the interest on natural flocculants has increased. The aminopolysaccharide chitosan and its modified derivatives have outstanding properties such as biocompability, biodegradability, hydrophilicity, adsorption, flocculating ability and antibacterial properties. These natural polymers derived from the sea-food industry waste products would be very useful as residue oil adsorbents in any oily wastewater and can be among the most promising candidates as a replacement of the synthetic flocculants. / Ölhaltiges Abwasser, das bei Erdöl- und petrolchemischen Raffinierungsprozessen entsteht, ist eine der größten Umweltgefahren. Dieses Altöl landet jedes Jahr in der Kanalisation und in Deponien. Es handelt sich dabei um die 25-fache Menge an Rohöl, die beim Unfall der Exxon Valdez (1989) ausgeflossen ist. Die Öl/Wasser-Trennung überspannt ein breites Spektrum industrieller Prozesse. Es gibt viele Techniken, die abhängig von jeder Situation eingesetzt werden. Das Nebenprodukt bei der Abtrennung von Wasser aus ölhaltigen Abwässern ist ein Schlamm (Ölschlamm), der reich an Öl, Tensiden und Partikeln ist. Der Ölschlamm enthält noch bedeutende Mengen an Wasser, die vor ihrer Entsorgung verwertet werden müssen. Die Verwendung von Polyelektrolyten zur Ausflockung des emulgierten Öls und seine Trennung von der wässrigen Phase ist in der Regel einer der Schritte zur Behandlung von Abwässern sowie ölhaltigen Schlämmen. Die Effizienz von Polyelektrolyten als Flockungsmittel wird ganz häufig über Versuch und Fehler bewertet, und das passende Polymer wird entsprechend dem jeweiligen Fall ausgewählt. Sogar in wissenschaftlichen Untersuchungen ist es eher üblich, industrielle Ölschlamm-Proben zu verwenden. Der industrielle Ölschlamm wird charakterisiert und mit Polyelektrolyten behandelt. Dennoch ist der industrielle Ölschlamm ziemlich kompliziert und variabel und muss durch ein Modell angenähert werden. Für die systematische Untersuchung der Effizienz von Polyelektrolyten als Flockungsmittel ist ein stabiles, realistisches und klar definiertes Ölschlamm-Modell notwendig. In der vorliegenden Arbeit wurde ein Ölschlamm-Modell erfolgreich entwickelt und charakterisiert. Das Modell besteht aus Wasser, Öl (Kerosin), Tensid (Natriumdodecylsulfat, SDS) und Tonteilchen (Blauton). Die Emulgiereffizienz des Tensids und des Tons wurden unabhängig voneinander untersucht. Die Wechselwirkungen zwischen dem Tensid und dem Ton, die sowohl die Adsorption des Ersteren auf dem Letzteren einschließen als auch einen Kationenaustausch, wurden untersucht. Die vier Komponenten des Ölschlamm-Modells wurden schließlich kombiniert und es wurde eine Reihe von Emulsionen hergestellt, bei denen die relativen Mengen der Emulsionsmittel verändert wurden, um eine möglichst hohe Stabilität zu erreichen. Nachdem ein geeignetes Ölschlamm-Modell zur Verfügung stand, wurde die Effizienz der verschiedenen Polyelektrolyte als Flockungsmittel bewertet. Kommerzielle natürliche (Chitosane), synthetische (PolyDADMACs) Polyelektrolyte und Oilbreaks sowie Labor-semi-synthetische Polymere (modifizierte Chitosane) wurden getestet. Die Flockungseffizienz wurde sowohl basierend auf der Menge und Qualität des Wassers, das zurückgewonnen wurde, als auch bezogen auf die Flockenstabilität, die Flockengröße und die Sedimentationsgeschwindigkeit bestimmt. Das zurückgewonnene Wasser wurde entsprechend der Vorschriften der Behörde für Umweltschutz (EPA) charakterisiert. Die Analysen enthielten die Bestimmung des gesamten organischen Kohlenstoffs (TOC), des chemischen Sauerstoffbedarf (CSB), des biochemischen Sauerstoffbedarfs (BSB5), des Gesamtgehalts an suspendierten Partikeln (TSS) und die Bestimmung des pH. Die Auswahl geeigneter Flockungsmittel hängt auch von der Art der gebildeten Flocken, in Kombination mit der Behandlung, die den Flockungsprozess folgt, ab. Wenn als Postflockungsprozess Filtration oder Zentrifugation folgen, sollten Polymere verwendet werden, die große und poröse Flöckchen bilden, wie im Fall der modifizierten Chitosane AG95 und AG97. Andererseits verlangen Geräte zur Klärung von Abwässern dichte Flocken, wie solche, die beim Einsatz von PolyDADMACs, kommerziellen Chitosanen, Oilbreaks und den modifizierten Chitosanen GA35, GA41 und AG79 entstehen. In Bezug auf die Wasserqualität erhält man mit einigen der verwendeten Polymere so niedrige Werte von CSB, TOC und BSB5, dass wie im Falle von P187K (PolyDADMAC) und GA41 (modifiziertes Chitosan) keine biologische Sekundärbehandlung notwendig ist. Im Falle der anderen Polymere ist eine biologische Behandlung nötig, um die vorgeschriebenen Grenzen zu erreichen. Die pH-Werte der modifizierten Chitosane (außer der AG97), der im Labor hergestellten PolyDADMACs und des Oilbreak OCAA sind alle in Bereich der vorgeschriebenen Grenzen. Die Anwendbarkeit von Biopolymeren als Flockungsmittel für die Ölschlammentwässerung ist ein relativ neuer Forschungsbereich. Als Folge der wachsenden Nachfrage nach umweltfreundlichen Technologien sowie erneuerbarer Ressourcen hat das Interesse an natürlichen Flockungsmitteln zugenommen. Das Aminopolysaccharidchitosan und dessen modifizierte Produkte haben hervorragende Eigenschaften wie Biokompatibilität, Biodegradierbarkeit, Hydrophilie, Adsorption, die Fähigkeit zur Flockung und antibakterielle Eigenschaften. Diese natürlichen Polymere, die aus Meeresfrüchte-Industrieabfallprodukten gewonnen werden, sollten als Restöl-Adsorbentien bei der Aufarbeitung jedes ölhaltigen Abwassers sehr nützlich sein und können zu den vielversprechendsten Kandidaten als Ersatz der synthetischen Flockungsmittel werden.