The isolation of DNA from plant tissue of the required quality is very complicated, especially because of the presence of substances that can interfere during amplification of DNA. These substances are mainly polyphenols, polysaccharides, proteins and various dyes. The chemical diversity of such materials can have a significant effect on the yield and quality of DNA using one isolation procedure. The main aim of the work was to evaluate the use of microisolation protocol for related matrices to the quality of the isolated DNA as well as the evaluation of the effect of inhibitors isolated with the nucleic acid to the amplification in the PCR. DNA was isolated from dried paprika (Capsicum annuum). In the first step, the samples were homogenized using a lysis reagent with cetyltrimethylammonium bromide. Subsequently, the DNA was purified by reversible adsorption on magnetic particles. It was tested six different modified particles. The concentration and purity of the obtained DNA was determined by spectrophotometry measuring the absorbance of the DNA solution in TE buffer. The quality of the DNA was confirmed by amplification in PCR. For the PCR were used primers specific for plant ribosomal DNA (rDNA). The presence of PCR products was detected by agarose gel electrophoresis. It was found out that used microassay is suitable for isolating of the DNA of the corresponding purity that is suitable for the genetic analysis by PCR. The differences were found between the magnetic particles that were tested for DNA isolation.

With a development of molecular biology methods it is an increasing interest in new procedures of DNA isolation of high quality. DNA isolation is performed on crude cell lysates by many techniques e.g. phenol extraction, salting out or adsorption on solid phase. Classical DNA isolation, such as phenol extraction is quite complicated and time consuming. New alternative methods of DNA isolation was development using reverse immobilizing DNA to a solid phase. Widespread is the use of the magnetic particles as carriers, which allow the isolation of DNA in high quality directly from crude cells lysates of complex samples. The current method of DNA adsorption onto the surface of magnetic particles does not provided sufficiently pure DNA for analysis of some comlex samples (e.g. food). Some inhibitors of the polymerase chain reaction (PCR) are apparently adsorbed onto the tube wall and the next step of DNA elution leads to their release into the solution and cpnsequent negative effect on quality of DNA (e.g. decreasing of PCR amplification). The principle of the developed procedure is design a device, which utilizes transfer of magnetic particles by paramagnetic newddle from one to another Eppendorf tube, in which further processing of the sample extends. Transfer of magnetic particles with DNA using needle prevents transmission of contaminating impurities. The proposed device allows to realize above-mentioned procedure. The functionality of the device being tested in the isolation of plasmid pUC19 DNA from crude lysates of E. Coli JM 109 (pUC19).

Micromethod of DNA isolation using magnetic particles is one of the modern technological methods used in DNA isolation, and makes the process simpler, more effective and faster. The main aim of this study was to isolate the DNA from various plant (tomato) food products, using different types of magnetic particles. The results were compared and the quantity, purity and the possibility of amplication of the isolated DNA among samples were found to be different. The DNA isolation method using magnetic particles P(HEMA-co-GMA) or HPS B-M-NH2 was shown to be the most effective in achieving the above mentiond parametres. DNAs from the analysed samples of plant food products were isolated in sufficient quantity and quality to be used in the conventional PCR. Differences in the possibility of the amplification of the isolated DNA stored at -20 °C during more than a half year were not found.

Carbon nanotubes (CNTs) have special characteristics that made them good agents especially for biomedical applications. In this study, Fe3O4 nanoparticles were incorporated onto the surface of CNTs followed by polyethylene glycol (PEG) grafting forming CNTs-Fe3O4-PEG hybrids. The PEGylation improves their biocompatibility, water solubility, and increases blood circulation. CNTs-Fe3O4-PEG was used as T2-contrat agent for magnetic resonance imaging (MRI). In addition, doxorubicin (DOX) was loaded onto CNTs-Fe3O4-PEG. The release of DOX from DOX-loaded CNTs-Fe3O4-PEG was tested under different pH conditions (7.4, 6.3 and 5.2). The release increased at acidic pH compared to neutral pH, which shows the sensitivity of the system to pH change. Triggering the release by Near Infra-Red (NIR) irradiation was furthermore investigated. The release increased after irradiation with NIR compared to control sample. These result prove that the designed system can be triggered by an internal stimuli (pH) and external stimuli (NIR irradiation) making it a promising candidate to be used for biomedical applications.

No description available.

Получены композиции на основе альгината натрия и фторкаучука СКФ-26, наполненные частицами магнитных материалов: Ni, Fe, Fe3O4, SrFe12O19, MQP-Системы изучены методами изотермической микрокалориметрии, механического динамического анализа и магнитометрии. Методом микрокалориметрии измерены энтальпии смешения компонентов композитных пленок и рассчитаны значения предельной энтальпии адгезии полимерных матриц к поверхности магнитных порошков. Показано, что для систем на основе СКФ-26 энтальпия адгезии является отрицательной величиной, абсолютные значения которой увеличиваются в ряду Ni-Fe-Fe3O4. В системах альгинат натрия/Ni и альгинат натрия/Fe реализуется атермическое смешение, а в системах альгинат натрия/ФС и альгинат натрия/ Fe3O4 – экзотермическое. С помощью метода ДМА получены концентрационные зависимости динамических модулей потерь и упругости, а также угла сдвига для систем на основе раствора альгината натрия и магнитных порошков Fe, Fe3O4 и SrFe12O19 при частоте 1 Гц и напряжении 1 Па. Обнаружено, что с увеличением концентрации частиц неорганического вещества в суспензиях наблюдается увеличение динамических модулей и уменьшение угла сдвига, причем для суспензии, содержащей магнетит, характер течения не меняется. При течении суспензий, содержащих порошки Fe или SrFe12O19 начинают преобладать упругие свойства. Методом магнитометрии для систем СКФ-26/Ni и CКФ-26/Fe были получены петли магнитного гистерезиса в диапазоне напряженности магнитного поля от -15 кЭ до 15 кЭ. Показано, что с ростом содержания порошка в композите остаточная намагниченность и намагниченность насыщения не меняются, а коэрцитивная сила уменьшается, т. е. на размагничивание магнитного материала требуется меньше энергии. Методом магнитометрии получены данные для систем на основе альгината натрия, содержащих 30% Fe или Fe3O4. Обнаружено, что для системы альгинат натрия/Fe значения намагниченности насыщения для композита больше, чем для индивидуального порошка Fe, что может быть обусловлено упорядочением спинов на поверхности магнитных частиц при формировании композита. / Compositions based on sodium alginate and SKF-26 fluoro-rubber filled with particles of magnetic materials: Ni, Fe, Fe3O4, SrFe12O19, MQP-systems were studied by isothermal microcalorimetry, mechanical dynamic analysis and magnetometry. The enthalpy of mixing of composite film components was measured by microcalorimetry and the values of the limiting enthalpy of adhesion of polymer matrices to the surface of magnetic powders were calculated. It is shown that for systems based on SKF-26, the enthalpy of adhesion is a negative value, the absolute values of which increase in the series Ni-Fe-Fe3O4. In the sodium alginate/Ni and sodium alginate/Fe systems, athermic mixing is realized, and in the sodium alginate/FS and sodium alginate/ Fe3O4 systems, exothermic mixing is realized. The DMA method is used to obtain the concentration dependences of the dynamic loss and elasticity modulus, as well as the shear angle, for systems based on a solution of sodium alginate and magnetic powders Fe, Fe3O4, and SrFe12O19 at a frequency of 1 Hz and a voltage of 1 Pa. It is found that with an increase in the concentration of inorganic matter particles in the suspensions, an increase in the dynamic modules and a decrease in the shear angle are observed, and for a suspension containing magnetite, the flow pattern does not change. During the course of suspensions containing Fe or SrFe12O19 powders, elastic properties begin to prevail. Magnetic hysteresis loops in the magnetic field strength range from -15 kE to 15 kE were obtained by magnetometry for the SKF-26/Ni and SKF-26/Fe systems. It is shown that with an increase in the powder content in the composite, the residual magnetization and saturation magnetization do not change, and the coercive force decreases, i.e., less energy is required to demagnetize the magnetic material. Data for systems based on sodium alginate containing 30% Fe or Fe3O4 were obtained by magnetometry. It is found that for the sodium alginate/Fe system, the saturation magnetization values for the composite are greater than for the individual Fe powder, which may be due to the ordering of spins on the surface of magnetic particles during the formation of the composite.

Este trabalho teve como objetivo sintetizar e caracterizar uma matriz híbrida estável de poli(estireno-co-divinilbenzeno) magnetizado pela adição de magnetita (Fe3O4) e avaliar seu potencial como suporte para a imobilização de lipases. A matriz híbrida foi sintetizado pela técnica de polimerização em suspensão utilizando dos monômeros de estireno e divinilbenzeno e ao qual foram adicionadas partículas de magnetita preparadas por coprecipitação dos íons Fe+2 e Fe+3. A caracterização foi realizada pelas técnicas de microscopia eletrônica de varredura (MEV), espectroscopia na região do infravermelho por transformada de Fourier (FTIR), difratometria de raios-X (DRX) e magnetização de amostra vibrante (VSM), comparando os materiais magnetizados e não magnetizados. Os biocatalisadores foram preparados pela imobilização da lipase de Candida rugosa (LCR) e lipase PS Burkholderia cepacia (LPS) via adsorção física e foram caracterizados em função da influência de pH e temperatura na atividade hidrolítica, parâmetros cinéticos, estabilidade térmica, estabilidade operacional e estabilidade de estocagem. O derivado de LCR foi aplicado em reações de esterificação e o derivado de LPS em reações de transesterificação. Os resultados obtidos pelas análises de FTIR, DRX e VSM confirmaram que a magnetita foi incorporada ao polímero, gerando atração das partículas por um campo magnético externo. A caracterização bioquímica indicou forte influência do pH na atividade hidrolítica, apresentando ponto ótimo próximo a 8,0 tanto para as lipases livres quanto imobilizadas. Os biocatalisadores magnetizados preparados apresentaram bom desempenho em todos os aspectos, o derivado da lipase de Candida rugosa alcançou conversões entre 89-94% nas reações de esterificação, revelando tempo de meia vida de t1/2=52 dias na estabilidade operacional. O derivado de Burkholderia cepacia atingiu rendimentos próximos a 80% nas reações de transesterificação com t1/2=40 dias. A imobilização aumentou a estabilidade térmica das lipases em 50 vezes no caso da LCR e em 2,3 vezes para a LPS. Estes resultados indicam que o material híbrido magnetizado sintetizado possui grande potencial para ser utilizado como suporte na imobilização de enzimas com aplicação em reações de interesse industrial. / This study aimed to synthesize and characterize a stable hybrid matrix of poly (styrene-codivinylbenzene) magnetized by the addition of magnetite (Fe3O4) and evaluate its potential for application in the immobilization of lipases, by characterization of the prepared biocatalysts. The support was synthesized by the suspension polymerization technique by applying styrene and divinylbenzene monomers and adding magnetite particles synthesized by co-precipitation of Fe + 2 and Fe + 3. The characterization of the material was performed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), vibrating sample magnetization (VSM), by comparison of the magnetized and the not magnetized particles. The biocatalysts were prepared by immobilization of lipase from Candida rugosa (CRL) and lipase from Burkholderia cepacia (Lipase PS) via physical adsorption and they were characterized according to the influence of pH and temperature on the hydrolytic activity, kinetic parameters, thermal stability, operational stability and storage stability. The CRL derivative was applied in esterification reactions and the lipase PS derivative was applied in transesterification reactions. The results obtained by the analysis FTIR, XRD and VSM confirmed the magnetite was successfully incorporated into the polymer and generated the atraction for an external magnetic field. Biochemical characterization indicated a strong influence of pH on the hydrolytic activity, showing better results on pH around 8,0 for free and both immobilized lipases. The magnetized biocatalysts prepared had good performance in all respects, derivative from Candida rugosa lipase reached 89-94% conversion in esterification reactions showing half-life of operational stability t1 / 2 = 52 days. The immobilized lipase from Burkholderia cepacia reached yields close to 80% in transesterification reactions presenting t1/2 = 40 days. Immobilization increased the thermal stability of lipase by 50 times in the case of CRL and 2,3 times for Lipase PS. These results indicate that the magnetized hybrid material synthesized has great potential to be used as a support for the immobilization of enzymes for use in reactions of industrial interest.

Este trabalho teve como objetivo sintetizar e caracterizar uma matriz híbrida estável de poli(estireno-co-divinilbenzeno) magnetizado pela adição de magnetita (Fe3O4) e avaliar seu potencial como suporte para a imobilização de lipases. A matriz híbrida foi sintetizado pela técnica de polimerização em suspensão utilizando dos monômeros de estireno e divinilbenzeno e ao qual foram adicionadas partículas de magnetita preparadas por coprecipitação dos íons Fe+2 e Fe+3. A caracterização foi realizada pelas técnicas de microscopia eletrônica de varredura (MEV), espectroscopia na região do infravermelho por transformada de Fourier (FTIR), difratometria de raios-X (DRX) e magnetização de amostra vibrante (VSM), comparando os materiais magnetizados e não magnetizados. Os biocatalisadores foram preparados pela imobilização da lipase de Candida rugosa (LCR) e lipase PS Burkholderia cepacia (LPS) via adsorção física e foram caracterizados em função da influência de pH e temperatura na atividade hidrolítica, parâmetros cinéticos, estabilidade térmica, estabilidade operacional e estabilidade de estocagem. O derivado de LCR foi aplicado em reações de esterificação e o derivado de LPS em reações de transesterificação. Os resultados obtidos pelas análises de FTIR, DRX e VSM confirmaram que a magnetita foi incorporada ao polímero, gerando atração das partículas por um campo magnético externo. A caracterização bioquímica indicou forte influência do pH na atividade hidrolítica, apresentando ponto ótimo próximo a 8,0 tanto para as lipases livres quanto imobilizadas. Os biocatalisadores magnetizados preparados apresentaram bom desempenho em todos os aspectos, o derivado da lipase de Candida rugosa alcançou conversões entre 89-94% nas reações de esterificação, revelando tempo de meia vida de t1/2=52 dias na estabilidade operacional. O derivado de Burkholderia cepacia atingiu rendimentos próximos a 80% nas reações de transesterificação com t1/2=40 dias. A imobilização aumentou a estabilidade térmica das lipases em 50 vezes no caso da LCR e em 2,3 vezes para a LPS. Estes resultados indicam que o material híbrido magnetizado sintetizado possui grande potencial para ser utilizado como suporte na imobilização de enzimas com aplicação em reações de interesse industrial. / This study aimed to synthesize and characterize a stable hybrid matrix of poly (styrene-codivinylbenzene) magnetized by the addition of magnetite (Fe3O4) and evaluate its potential for application in the immobilization of lipases, by characterization of the prepared biocatalysts. The support was synthesized by the suspension polymerization technique by applying styrene and divinylbenzene monomers and adding magnetite particles synthesized by co-precipitation of Fe + 2 and Fe + 3. The characterization of the material was performed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), vibrating sample magnetization (VSM), by comparison of the magnetized and the not magnetized particles. The biocatalysts were prepared by immobilization of lipase from Candida rugosa (CRL) and lipase from Burkholderia cepacia (Lipase PS) via physical adsorption and they were characterized according to the influence of pH and temperature on the hydrolytic activity, kinetic parameters, thermal stability, operational stability and storage stability. The CRL derivative was applied in esterification reactions and the lipase PS derivative was applied in transesterification reactions. The results obtained by the analysis FTIR, XRD and VSM confirmed the magnetite was successfully incorporated into the polymer and generated the atraction for an external magnetic field. Biochemical characterization indicated a strong influence of pH on the hydrolytic activity, showing better results on pH around 8,0 for free and both immobilized lipases. The magnetized biocatalysts prepared had good performance in all respects, derivative from Candida rugosa lipase reached 89-94% conversion in esterification reactions showing half-life of operational stability t1 / 2 = 52 days. The immobilized lipase from Burkholderia cepacia reached yields close to 80% in transesterification reactions presenting t1/2 = 40 days. Immobilization increased the thermal stability of lipase by 50 times in the case of CRL and 2,3 times for Lipase PS. These results indicate that the magnetized hybrid material synthesized has great potential to be used as a support for the immobilization of enzymes for use in reactions of industrial interest.

Inspirados por las investigaciones realizadas en los campos de la plasmónica y los metamateriales, en este trabajo hemos estudiado la difusión de la luz por pequeñas partículas (nanopartículas en el rango del visible) con propiedades ópticas arbitraria, tanto convencionales (dieléctricas como metálicas) como no convencionales (con permeabilidad magnética distinta de 1). El trabajo está principalmente enfocado en controlar la propiedades (principalmente la dirección) de la luz difundida por una partícula mediante la manipulación de sus constantes ópticas. Las principales aplicaciones de este estudio se pueden concentrar en los campos de los biosensores y/o de las comunicaciones ópticas. Es por esto que hemos considerado tanto partículas aisladas como sistemas de partículas(dímeros principalmente). / Inspired by the last researches on plasmonics and metamaterials, this work is devoted to the study of light scattering by small particles (nanoparticles in the visible range) with arbitrary optical properties, both conventional and non-conventional. We focused the analysis on the control of the directionality of light scattering by tunning the optical constants of the scatterer. This could be interesting for the design of futuristic optical communications and/or for the generation of improved biosensores. For this reason, isolated particles and also clusters of them (mainly dimers) were considered.

Submitted by MARCO ANTONIO BAPTISTELLA null (marco_baptistella@terra.com.br) on 2016-08-23T19:37:34Z No. of bitstreams: 1 REV 17. MARCO A. BAPTISTELLA.pdf: 11920777 bytes, checksum: ba33aea22870d52b190f07f07f6e333c (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-08-25T20:22:58Z (GMT) No. of bitstreams: 1 baptistella_ma_me_guara.pdf: 11920777 bytes, checksum: ba33aea22870d52b190f07f07f6e333c (MD5) / Made available in DSpace on 2016-08-25T20:22:58Z (GMT). No. of bitstreams: 1 baptistella_ma_me_guara.pdf: 11920777 bytes, checksum: ba33aea22870d52b190f07f07f6e333c (MD5) Previous issue date: 2016-08-04 / Atualmente, as cargas aplicadas nos eixos de vagões ferroviários têm sofrido aumento de esforços devido ao choque e tração no engate ferroviário. Neste trabalho o estudo foi realizado no engate tipo “F”, que tem maior utilização e interesse por parte da ferrovia, por se tratar de um engate rotativo e utilizado para descarregamento de vagões com os mais diversos tipos de carga. Este trabalho tem como objetivo, verificar se existem transformações da microestrutura no material do engate, e possíveis gerações de trincas, através do estudo do aço AAR M-201 Grau E, após o ensaio de partículas magnéticas, com corrente alternada através da aplicação de Prods. A faixa de amperagem variou-se de 100 a 1000 ampères e como meio de resfriamento, utilizou-se a água a temperatura ambiente e o ar forçado. A preparação foi feita partindo de um engate Tipo “F” completo, fornecido por um operador logístico, a cauda foi magnetizada por inteiro e depois separados os corpos de prova por corte de serra, refrigerado para não haver aquecimento. Para a caracterização microestrutural, utilizou-se uma preparação metalográfica de laboratório de bancada e ataque com Nital 2%, e fez-se uso de microscopia ótica convencional e microscopia eletrônica de varredura (MEV). Através da utilização da microscopia ótica e da microscopia eletrônica de varredura (MEV), pode-se verificar a microestrutura nas áreas que sofreram alterações microestruturais. Concluiu-se que o aço do engate, AAR M-201 grau E, após a magnetização por Prods de 300 a 1000 ampères, apresentou alteração microestrutural, onde observou-se uma estrutura de martensita acicular não revenida e geração de trincas, a mesma alteração microstrutural foi observada em amostras com resfriamento ao ar, porém sem a geração de trincas, em ampliações de 50 vezes foi nítido o efeito de queima provocado pelo Prods no material, bem como sua extensão, evidenciando áreas centrais de atuação do Prods, zonas de transição e áreas não afetadas. As trincas tiveram comportamento de propagação intergranular, para ampliações de 9300 vezes em MEV, pode-se observar que as mesmas se propagaram somente nas áreas com martensita e contornaram as áreas com maior presença da fase ferrítica. / Currently, the applied loads on the axles of railway cars has been increasing efforts due to shock and traction in railway coupling. In this work, the study was performed on the coupling type "F", which has increased use and interest on the part of the railway, because it is a rotary coupling and used for unloading of wagons with the most diverse types of cargo. This work aims, check for microstructural changes on the coupling material, and possible generation of cracks through the study of steel AAR M-201 grade and after the test of magnetic particles with alternating current by applying Prods. As amperage range, was varied from 100 to 1000 amps and as cooling medium water was used at room temperature and forced air. The preparation was made starting from a coupling type "F" full, provided by a logistics operator, the tail was magnetized in full and then separate the specimens by cutting saw, cooled to no heating. For microstructural characterization, we used a metallographic preparation of benchtop laboratory and attack Nital 2%, and was made use of conventional optical microscopy and scanning electron microscopy (SEM). By using light microscopy and scanning electron microscopy (SEM), you can check the microstructure in areas that suffered microstructural changes. It was concluded that the coupling of steel, AAR M-201 grade And after magnetization by Prods 300-1000 amps, presented change microstructural where there was a martensite acicular structure not tempered and generation of cracks, the same change microstructural was observed in air to cooling with samples, but without generating cracks at magnifications of 50 times was clear the burning effect caused by Prods in the material as well as its extension, showing the central areas of the Prods activities, transition zones and unaffected areas. The intergranular crack propagation behavior had to 9300 times magnification under SEM, it can be seen that they have spread only in areas with martensite and rounded areas with the greatest presence of the ferritic phase.