A abóbora se destaca como uma hortaliça de grande potencial nutritivo e tecnológico. Para que se possa aumentar a variabilidade dos produtos de origem vegetal disponíveis no mercado, a abóbora deve ser submetida a processos térmicos que visam prolongar a sua vida de prateleira, além de realizar as modificações necessárias para a sua comercialização. Neste contexto, o objetivo principal da presente pesquisa foi avaliar a aplicação da tecnologia de aquecimento ôhmico em diferentes etapas a serem realizadas durante o processamento da abóbora híbrida Tetsukabuto. Na primeira etapa do trabalho, objetivou-se investigar os efeitos do branqueamento convencional e ôhmico (realizados a 80 ºC durante 4 minutos e 163 V no aquecimento ôhmico) sobre a inativação da enzima peroxidase e sobre a cor das abóboras. Para o ajuste dos dados relativos à inativação enzimática, diferentes modelos cinéticos foram avaliados, sendo o modelo de distribuição de Weibull o escolhido para descrever a cinética de inativação da peroxidase. Com este estudo, foi possível observar que o branqueamento com campo elétrico afetou significativamente os parâmetros cinéticos de inativação enzimática, tornando esse processo mais rápido. A aplicação de ambas as tecnologias não causou modificações significativas dos parâmetros de cor da abóbora. Na segunda etapa do trabalho, creme de abóbora foi pasteurizado via tratamento térmico convencional e ôhmico realizados a 90 ºC por 2 horas e 47 V no aquecimento ôhmico. O objetivo dessa etapa foi avaliar os efeitos de ambas as tecnologias sobre a variação de cor, degradação de carotenoides totais e mudanças nos parâmetros reológicos do creme. Foi possível observar que a cor do creme foi levemente afetada durante a aplicação dos processos, tornando-se ligeiramente mais escura e perdendo o tom amarelado. Além disso, foi verificada perda no teor de carotenoides totais presentes no creme. Ainda, não foi observado um efeito não-térmico adicional do campo elétrico sobre a cor e os carotenoides presentes no produto. O creme de abóbora apresentou comportamento pseudoplástico e que se ajustou satisfatoriamente ao Modelo da Lei da Potência. Com a aplicação dos tratamentos, foi observado um aumento significativo no índice de consistência do creme, sendo que ambas as tecnologias não apresentaram diferenças representativas entre si. O conjunto de resultados obtidos pelo presente trabalho sugere que o aquecimento ôhmico é uma tecnologia promissora a ser aplicada em diferentes operações envolvidas durante o processamento de alimentos. / Pumpkins stand out as vegetable of great nutritional and technological potential. In order to increase the variability of vegetal products available on the market, pumpkins must be subjected to thermal process which can prolong its shelf life, and imposing also the necessary modifications for its commercialization. In this context, the main objective of the present research was to evaluate the application of ohmic heating technology in different stages carried out during the processing of the hybrid pumpkin Tetsukabuto. In the first part of the work, the objective was to investigate the effects of conventional and ohmic blanching (performed at 80 ºC during 4 minutes and 163 V in the ohmic heating) on the inactivation of peroxidase enzyme and on the color of the pumpkins. In order to adjust the data related to enzyme inactivation, different kinetic models were evaluated, being the Weibull distribution model chosen to describe the inactivation kinetics of peroxidase. With this study, it was possible to observe that the electric field blanching significantly affected the kinetic parameters of enzymatic inactivation, turning this process faster. The application of both technologies did not cause significant modifications of pumpkins color parameters. In the second part of the work, pumpkin cream was pasteurized via conventional and ohmic thermal treatment carried out at 90 ºC for 2 hours and 47 V in the ohmic heating. The objective of this stage was to evaluate the effects of both technologies on the color variation, the degradation of total carotenoids and the changes in rheological parameters of the cream. It was possible to observe that the color of the cream was slightly affected during the application of the processes, becoming slightly darker and losing its yellowish tone. In addition, it was verified loss of total carotenoid content in the cream. Furthermore, no additional non-thermal effect of the electric field on the color and on carotenoids present in the product was observed. The pumpkin cream presented a shear thinning behavior and was satisfactorily adjusted to the Power Law Model. It was observed a significant increase in the consistency index of the cream after the application of both treatments, and both technologies did not present significative differences between them. The overall results obtained in the present work suggests that ohmic heating is a promising technology to be applied in different operations involved during food processing.

Made available in DSpace on 2014-06-11T19:22:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-22Bitstream added on 2014-06-13T19:48:27Z : No. of bitstreams: 1 panoff_b_me_botfca.pdf: 479570 bytes, checksum: 18963582936b7c7dfc0dd966e4523c3d (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Devido ao aumento do número de cultivares de soja e com pouca diferença genética entre elas, fica cada vez mais difícil a verificação de contaminação varietal pelo método de peroxidase. As cultivares de soja são separadas em dois grupos com base na atividade, alta ou baixa, da peroxidase no tegumento. A alta atividade é resultado da presença de, pelo menos, um alelo dominante (EpEp ou Epep), enquanto que baixa atividade resulta da presença do par recessivo (epep). O auxílio de técnicas moleculares na identificação de contaminação varietal de soja utilizando a peroxidase é de grande importância, pois a tecnologia baseada na análise do DNA não sofre a ação de fatores externos (ambientais). Neste contexto, o objetivo geral do trabalho foi o de viabilizar a técnica de PCR convencional para verificar a presença de contaminação varietal em auxílio ao método colorimétrico da peroxidase. O estudo foi conduzido no Departamento de Produção Vegetal da Faculdade de Ciências Agronômicas/UNESP, Campus de Botucatu-SP. Foi realizado o teste colorimétrico tradicional e comparado com os resultados encontrados no PCR. Foram usadas 14 cultivares de importância, dessas, foram pré-selecionadas seis cultivares com reação positiva à peroxidase: BRS 320, BRS 284, BRS 232, BRS 7860RR, BRSMG 760SRR, BRS295RR, quatro cultivares negativas à peroxidase: BRS 326, BRS 8160RR, Nutrisoy, BRS Valiosa RR e quatro cultivares com reação positiva e negativa: BRSGO 8060, BRS 270RR, FTS Campo Mourão e BRS 239. Em paralelo ao teste colorimétrico, o DNA foi extraído das sementes inteiras, primers foram desenhados, seguido da reação de PCR e eletroforese em gel de agarose. A utilização de kit comercial de extração de DNA, juntamente com a utilização da técnica de PCR foi eficiente na detecção de amostras de sementes... / The increasing number of soybean cultivars and the small genetic difference between makes difficult to verify varietal contamination using the conventional peroxidase method. The soybean cultivars are separated into groups with high or low activity, based on the peroxidase activity in the tegument,. The high activity is a result of the presence of at least one dominant allele (or EpEp Epep), while low activity results from the presence of the pair recessive (epep). The use of molecular techniques to identify soybean cultivars is a powerful tool since it is not influenced by external factors (environmental) or genetic. In this context, the general objective of this work was to enable the use of molecular biology technique to facilitate identification of soybean cultivars. The study was performed at the Department of Plant Production, Faculty of Agricultural Sciences / UNESP, Botucatu-SP. Traditional biochemical colorimetric test was performed and compared with the results of obtained by conventional PCR. Fourteen commercial soybean cultivars were used, where we selected six cultivars with positive reaction to peroxidase: BRS 320, BRS 284, BRS 232, BRS 7860RR, BRSMG 760SRR, BRS295RR, four cultivars with negative reaction to peroxidase: BRS 326, BRS 8160RR, NutriSoy, BRS Valiosa RR and four cultivars with positive and negative reaction: BRSGO 8060, BRS 270RR, FTS Campo Mourão and BRS 239. In parallel with the traditional colorimetric assay for peroxidase, DNA was extracted from whole seeds and primers were designed, followed by conventional PCR reactions and visualization in agarose gel. The use of commercial kit for DNA extraction along with the use of the PCR technique was efficient in detecting negative and positive samples. However, when seed lots with positive reaction was purposely contaminated seed lots with negative reaction at the proportion of 1, 2, 3, 4 and 5%, the PCR technique was not sensible enough to detect the contamination

Thyroid carcinoma is the most common endocrine malignant neoplasia. Differentiated thyroid carcinomas (DTC), represent more than 90% of all thyroid carcinomas and comprise the papillary (PTC) and follicular thyroid carcinomas (FTC) subtypes. Anaplastic thyroid carcinoma (ATC) corresponds to less than 5% of all thyroid tumors. The etiology of DTC is not fully understood. Several genetic events have been implicated on differentiated thyroid tumorigenesis. Point mutations in BRAF and RAS genes and RET/PTC rearrangements are observed in about 70% of PTC cases. Follicular carcinomas commonly harbor RAS mutations and PAX8-PPARJ rearrangements. Anaplastic carcinomas may harbor a wide set of genetic alterations, as in genes encoding effectors in the mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinases (PI3K) and G-Catenin signaling pathways. These distinct genetic alterations are able to activate constitutively several signaling pathways as MAPK, PI3K and G-Catenin, which have been implicated on thyroid cancer development and progression. In this context, the evaluation of the specific oncogenes, as well as the knowledge of their effects on thyroid carcinomas can provide important information about the disease presentation, prognosis and therapy, through the development of specific tyrosine kinase targets. Particularly in this review, we explore the main genetic alterations observed in follicular cell-derived thyroid carcinomas as well as the molecular mechanisms involved in thyroid tumors development and progression.

Orientador: Humberto Tonhati / Coorientador: Daniele Fernanda Jovino / Coorientador: Larissa Fernanda Simielli Fonseca Noronha / Banca: Marcia Cristina de Sena Oliveira / Banca: Rodrigo Pelicioni Savegnago / Resumo: A mastite é a principal doença que acomete os rebanhos leiteiros e promove grandes perdas na produção devido a alterações na composição do leite e prejuízos na saúde do animal, aumentando assim os custos de produção. Caracteriza-se pela resposta inflamatória da glândula mamária a agentes infecciosos, alterando sua fisiologia e metabolismo. Estudo de expressão gênica vem sendo empregado em vários organismos com a finalidade de identificar genes relacionados com características economicamente importantes como resistência a doenças. Com o objetivo de compreender melhor os mecanismos de resposta imune envolvidos no fenótipo de resistência/susceptibilidade à mastite, foi realizada a quantificação da expressão relativa dos genes do Fator Estimulador de Colônias de Granulócitos 3 (CSF3) e Lactoperoxidase (LPO), em leite de búfalas mestiças saudáveis e com mastite, por meio da técnica de PCR quantitativa em tempo real (qRT-PCR). Para isso, foi realizada a extração de RNA total, com Trizol, de células somáticas do leite de doze animais diagnosticados com mastite e de doze animais livres da infecção. Foram realizadas análises de qualidade e quantificação do RNA total e das contaminações por DNA genômico para garantir a integridade das amostras. A quantificação da expressão gênica foi realizada por meio do método do Delta Delta Ct. Os genes GAPH, HPRT1, EEF1A1 e RPLP0, foram selecionados na literatura, e testados como genes de referência. As análises referentes às quanti... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Not available / Mestre

<p>In silico ligand docking is a versatile and common technique when predicting ligands and inhibitors for protein binding sites. The various docking programmes aim to calculate binding energies and to predict interactions, thus identifying potential ligands.The currently available programmes lack satisfying means by which to account for structural water molecules which can either mediate protein-ligand contacts or be displaced upon ligand binding. The present project aims to generate data to facilitate the global work of developing scoring functions in docking programmes to account for structural water molecules contribution to ligand binding to fill the said void. This is done by validating the performance of docking using a simple model system (cytochrome C peroxidase (CCP) W191G) containing four well ordered, deeply buried structural water molecules which are known to either interact with a ligand or to be displaced upon ligand binding.Known ligands were docked into eight (crystallographically determined) receptor set-ups comprising the receptor and no, one or two of the water molecules. The performance was validated by comparison of the binding modes of the docked ligands and the crystal structures, comparison of docking scores of the ligands in the different set-ups, enrichment of the ligands from a database of decoys and finally by predicting new ligands from the decoy database. In addition a high resolution crystal structure of CCP W191G in complex with 3-aminopyridine (3AP) was determined in order to resolve ambiguities in the binding mode of this ligand.</p>

Thesis (doctoral)--Swedish University of Agricultural Sciences, 2004. / Thesis documentation sheet inserted. Errata sheet inserted. Appendix reprints four papers and manuscripts co-authored with others. Includes bibliographical references. Also issued electronically via World Wide Web in PDF format; online version lacks appendix.

Phenols are priority pollutants that are commonly found in a large number of industrial wastewaters. Different processes are currently available for the elimination of phenol from wastewater but present some disadvantages like low efficiency, high energy-consumption, the necessity of acclimatisation of the sludges or the limitation of the treatment capacity. The need to find alternatives has made the enzymatic processes a good option. In the last two decades, several processes were implemented with different enzymes from plants and microorganisms, including peroxidases from several sources, as the horseradish peroxidase.Also, different enzyme configurations, free or immobilised enzyme and different supports for immobilisation have been studied. Substantial attention has been devoted to the covalent immobilisation of enzymes on porous insoluble supports such as glass, alumina, silica, and chitosan.The main novelty of this work is the utilisation of a torus reactor for the removal of organic contaminants from wastewaters. This reactor, which can be considered as a loop reactor, presents some advantages over other stirred tank reactors.The goal of this work is the study of the hydrodynamics of a torus reactor for its further application in the enzymatic elimination of phenol and the coupling of the kinetics and the modelisation.In a first step, the enzymatic elimination of phenol was experimentally studied in the torus reactor. In order to compare the performances, several assays were also carried out with a stirred reactor. A high degree of conversion was obtained for the enzymatic elimination of phenol in both reactors with the tested quantities of phenol. It was concluded that, keeping a ratio of 1:1 between the phenol and the H2O2 initial molar quantities, the highest final reaction conversion was obtained. Using the torus reactor was obtained 97% of phenol conversion when the optimal concentrations of substrates were usedIn order to improve economically the process, the enzyme should be used in a continuous regime over a long time period to exploit it completely. For this reason it was necessary to immobilise the enzyme. This work presents a new configuration that has never been tested: the horseradish peroxidase supported on Eupergit.In a second step, the characterisation using the CFD of the flow-field in a torus reactor of 100 ml, similar to the experimental reactor, was carried out for two different configurations, batch and continuous operating modes. Moreover, the scale-up of the volume of the torus reactor was carried out using CFD for a 300 ml reactor.Finally, the enzymatic reaction of phenol with the HRP was modelled using the CFD coupled to the kinetic model of the enzymatic reaction to the flow simulation. These results allowed the possibility of optimising and scaling-up the process using the CFD modelisation. / Los compuestos fenólicos son contaminantes prioritarios que se encuentran comúnmente en una gran cantidad de efluentes industriales. Diferentes procesos están disponibles actualmente para la eliminación de fenol desde dicho efluentes pero los mismos presentan algunas desventajas como pueden ser una baja eficiencia, un mayor consumo de energía, la producción de lodos conteniendo hierro o limitaciones en la capacidad de tratamiento. La necesidad de encontrar alternativas a estos problemas ha hecho del proceso enzimático una buena opción. En las últimas dos décadas, varios procesos han sido implementados utilizando diferentes enzimas extraídas de plantas y microorganismos como pueden ser las peroxidasas de diversas fuentes, incluyendo la horseradish peroxidasa.Diferentes configuraciones de enzimas, libre e inmovilizada y diferentes soportes para la inmovilización han sido también estudiados. Sustancial atención ha sido dedicada a la inmovilización de enzimas por enlace covalente sobre soportes porosos insolubles tales como vidrio, aluminio, sílice y chitosan. El objetivo de este trabajo es el estudio de la hidrodinámica dentro de un reactor tórico para su posterior aplicación en la eliminación enzimática de fenol y el acople entre las cinéticas y la modelización.En una primera etapa, la eliminación enzimática de fenol es estudiada experimentalmente en el reactor tórico. Con el objetivo de comparar el rendimiento de dicho reactor, varios ensayos se realizaron en un reactor agitado tradicional. Un alto grado de conversión de fenol ha sido obtenido para la eliminación enzimática de fenol en ambos reactores para las cantidades estudiadas de fenol. Ha sido observado que es necesario mantener una relación de 1:1 entre la concentración inicial de fenol y la de peróxido de hidrógeno para lograr la mayor conversión de fenol. Usando el reactor tórico ha sido obtenido un 97% de conversión de fenol cuando las concentraciones óptimas de substratos y enzimas fueron utilizados. Con el objetivo de mejorar económicamente el proceso y hacerlo factible para su uso a escala industrial, la enzima debería ser utilizada en un proceso en continuo sobre un largo período de tiempo para explotarla completamente. Por esta razón, ha sido necesario inmovilizar la enzima. Este trabajo muestra una nueva configuración que no ha sido aún probada: la horseradish peroxidase soportada en Eupergit. Asimismo, la caracterización usando CFD del campo de flujo de un reactor tórico similar al experimental de 100 ml ha sido realizada para un reactor trabajando de forma batch y continua. Un escalado en el volumen del reactor tórico ha sido realizado utilizando CFD para un reactor de 300 ml. Finalmente, la reacción enzimática de fenol con HRP has sido modelada acoplando el modelo cinético obtenido experimentalmente con las simulaciones del campo de flujo dentro del reactor. Estos resultados permitirán la optimización y el escalado del proceso usando CFD.

Infection of fully expanded leaves of tobacco with INF1 causes the appearance of HR lesions within 12 h and progressive to all infection sites after 48 h treatment. Among the POD isozymes, the increase of cationic PODs and anionic PODs is correlated with the rise of lignin contents in INF1-treated leaves, especially cationic PODs (pI 9.5, pI 8.7, pI 8.3, pI 7.8, pI 7.4). It was suggested that the induction of POD activity resulted in part of H2O2 reduction. The increase of cationic (pI 9.5) and anionic (pI 4.4) POD transcripts was correlated with the increased cationic and anionic PODs activity in INF1-treated leaves. Therefore, the increased POD activity is due to the de novo synthesis of the cationic (pI 9.5) and anionic (pI 4.4) PODs in INF1-treated leaves. The increase in cationic pI 9.6 laccase transcript was also correlated with the increased cationic laccase activity in INF1-treated leaves. Our results suggest that laccase might play a major role on lignin biosynthesis at the early stage (6 h), and as the inoculation time was prolonged, peroxidases (especially cationic POD) and laccases will work together on lignin biosynthesis.

Cu-treated peanut seedlings not only showed a significant inhibition in peanut root growth, but also a decrease in endogenous indole-3-acetic acid (IAA) contents . The decline of IAA content in Cu-treated tissues is accompanied by an increase in cationic peroxidase activity. In addition, the increase in cationic peroxidase (pI 8.5) activity is correlated with an increase in cationic peroxidase transcripts. Cu might suppress the growth of peanut roots by inducing the synthesis of the cationic peroxidase isozyme that degrades endogenous IAA, causing the retardation of peanut root growth. The increase in activity of anionic peroxidases (pI 3.5) is also correlated with the rise in lignin content in Cu-treated roots. We suggest that the increase in anionic peroxidases (pI 3.5) induced by Cu might be responsible for lignin synthesis in peanut roots, and remove excess hydrogen peroxide, thus serving a detoxifying role during Cu treatment.

¡iAbstract¡j Vagal sensory neurons play an important role in the neural control of airway and other visceral organs. Regional distribution of vagal sensory neurons in the vagal ganglia that correspond to different viscera is uncertain. Horseradish peroxidase (HRP) was applied to the right thoracic vagus nerve and recurrent laryngeal nerve to be retrogradely transported to the neurons in vagal nodose and jugular ganglia. Labeling of neuronal cell bodies was visualized with diaminobenzidine reaction. Ricinus communis agglutinin-60 (RCA-60) was injected into the right thoracic vagus nerve and transported retrogradely to cause destruction of the ribosome in the vagal ganglia neuron cell bodies. The magnitude of neurogenic plasma extravasation induced by capsaicin was measured by the area density of the India ink-labeled leaky blood vessels in the trachea and bronchi. The present study demonstrated that there was no distinct localization of HRP- labeled neurons, except at the level of pharyngeal nerve that was connected to the middle of the nodose ganglion. There were approximately 42.2 % of labeled neurons in the nodose ganglion and 30.5 % in the jugular ganglion 3 days after application of HRP in the thoracic vagus nerve. After application of HRP to the recurrent laryngeal nerve, 13.3 % of labeled neurons was found in the nodose ganglion and 8.3 % in the jugular ganglion. One to two weeks after RCA injection into the thoracic vagus nerve, many cell bodies of neurons had a striking degenerative alteration and the cytoplasmic density was markedly reduced. Nissl¡¦s bodies obviously disappeared and vacuoles were the usual feature. Application of RCA-60 also inhibited the neurogenic plasma extravasation in the right bronchial tree that were reduced by 71- 89 %. It is suggested that RCA- 60 selectively destroyed the vagal sensory neurons that innervated the ipsilateral branchial tree. It is concluded that vagal nodose and jugular ganglia supplied different number of sensory neurons to the vagal branches and regulated the physiological function of the visceral organs.