Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-10T14:31:41Z No. of bitstreams: 1 Vania Freire Lemos.pdf: 1507844 bytes, checksum: cbb75eb979bd39586c08dadf5aea6579 (MD5) / Made available in DSpace on 2017-02-10T14:31:41Z (GMT). No. of bitstreams: 1 Vania Freire Lemos.pdf: 1507844 bytes, checksum: cbb75eb979bd39586c08dadf5aea6579 (MD5) Previous issue date: 2013-02-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The objective of this work was to evaluate dynamics of the proteinogram of blood serum and whey protein of Santa Ines sheep breed following the antipartum period and during lactation, and to compare/to quantify proteins detected at eletrophoresis of the whey protein from healthy and infectious mammary glands in different phases of lactation. Thrirty four sheeps submitted to half-intensive system with same sanitary and nutritional management has been followed. For accomplishment of proteinogram of the sheep, they had been investigated during approximately 10 days from antipartum and 15, 30, 60 and 90 days postpartum, moments where clinical examination of mammary gland was carried through. Blood serum was evaluated at antipartum moment and proteinogram of whey protein at subsequent moments. Bacteriological culture and biochemist characterization of milk samples for confirmation of healthy and infected glands was performed. Separation of protein fractions was carried through using sodium dodecil sulphate polyacrylamide gel eletrophoresis (SDS-PAGE). For the blood serum it was observed quantification of nine proteins with significant influence in IgG. At whey protein, influence of the phases of lactation was identified eigth proteins, having albumin, IgG and β - lactoglobulin.. Comparing healthy and infected glands it was verified that hatptoglobin and α-1 acid glycoprotein, lactoferrin, albumin and immunoglobulins IgA and IgG in whey protein act as potentials biomarkers of infection in mammary gland of ovine species. / Objetivou-se neste trabalho avaliar a dinâmica do proteinograma do soro sangüíneo e lácteo de ovelhas da raça Santa Inês acompanhadas no período que antecedeu o parto e durante a lactação e comparar/quantificar as proteínas detectadas no traçado eletroforético do soro lácteo de glândulas mamárias sadias e infectadas em diferentes fases da lactação. Foram acompanhadas 34 ovelhas submetidas ao sistema de criação semi-intensivo, com mesmo manejo higiênico, sanitário e nutricional. Para a realização do proteinograma as ovelhas foram investigadas durante, aproximadamente 10 dias que precedeu o parto e 15, 30, 60 e 90 dias após o parto, momentos em que foi realizado o exame clínico da glândula mamária. O proteinograma sangüíneo foi efetuado a partir do momento pré parto e o proteinograma do soro lácteo nos momentos subseqüentes. Realizou-se o cultivo bacteriológico e a caracterização bioquímica das amostras de leite para confirmação de glândulas sadias e infectadas. A separação das frações protéicas foi realizada utilizando-se eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Para o soro sanguíneo observou-se a quantificação de nove proteínas observando influência significativa somente na IgG; no soro lácteo identificou-se oito proteínas havendo influência das fases de lactação na albumina, IgG e β - lactoglobulina. Ccomparando glândulas sadias e infectadas verificou-se que a hatptoglobina, α-1 glicoproteína ácida, lactoferrina, albumina e as imunoglobulinas IgA e IgG presentes no soro lácteo atuam como biomarcadores de infecção na glândula mamária na espécie ovina.

1. For the first time a wide range of biochemical genetic markers and different typing techniques were used to access levels of genetic variability in Lithuanian horse breeds; 2. DNA based methods were used to access levels of genetic variation in Lithuanian horse breeds; 3. Genetic variation in Lithuanian horses was investigated using mitochondrial DNA (mtDNA) sequencing; 4. Genetic relationship and genetic distances between the breeds were estimated using a wide range of different genetic markers.

Submitted by Mery Piedad Zamudio Igami (mery@ipen.br) on 2017-03-10T15:01:44Z No. of bitstreams: 1 22058.pdf: 1629255 bytes, checksum: 26070b2e80c7a4ee0c8281ffabb5a414 (MD5) / Made available in DSpace on 2017-03-10T15:01:44Z (GMT). No. of bitstreams: 1 22058.pdf: 1629255 bytes, checksum: 26070b2e80c7a4ee0c8281ffabb5a414 (MD5) / As raias são peixes peçonhentos e estão frequentemente associadas a acidentes em seres humanos, principalmente na região Norte do Brasil, favorecidos pelo hábito desses peixes de permanecerem no fundo de águas rasas e pela contínua utilização humana dos rios. Os ferrões das raias causam lesões dolorosas, edema, necrose, e o muco que recobre toda a extensão do corpo desses peixes pode aumentar a gravidade desses ferimentos. O objetivo deste trabalho foi avaliar a resposta imunológica induzida pelo muco de Paratrygon aiereba nativo ou irradiado por raios gama de 60Co em modelos animais. Foram realizados ensaios imunoenzimáticos e Western blotting para verificar a resposta humoral e reatividade cruzada dos soros provenientes de camundongos Swiss e coelhos New Zealand previamente imunizados contra o veneno, muco nativo ou irradiado. A indução da produção de anticorpos in vitro, as subclasses de IgG e a quantificação de citocinas foram analisados. Além de realizados ensaios de soroneutralização da atividade edematogênica in vitro e in vivo e de viabilidade celular. Os dados foram analisados estatisticamente por meio de análise de variância. O protocolo de imunização possibilitou a obtenção de soros com títulos satisfatórios de anticorpos policlonais. O muco e veneno de P. aiereba são imunogênicos e apresentam reatividade antigênica. O muco nativo ou irradiado induziu a produção de anticorpos IgG e esses reconheceram antígenos presentes no muco de outras espécies de raias. Células esplênicas de animais imunizados contra o muco irradiado produziram IFN-γ, TNF- α e IL-10 e também foi observada a produção sérica de TNF-α (grupo imunizado contra o muco irradiado) e de IL-6 e IL-17 (grupo imunizado contra o muco nativo). O soro anti-muco irradiado reduziu a atividade edematogênica in vitro, ao contrária da in vivo que não foi neutralizada. Os resultados corroboram o uso da radiação ionizante, com produção de anticorpos altamente responsivos e melhor resposta imune, além de comprovar que o muco de Paratrygon aiereba foi capaz de estimular resposta imune adaptativa celular e humoral. / Tese (Doutorado em Tecnologia Nuclear ) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Alzheimer disease (AD) is the most frequent dementia. The prevalence is approximately 10% in 65 years old people. The current treatment is only progression protective, therefore it is crucial to find a new diagnostic approach for diagnosing AD in early stage. We analysed a set of 55 patients by the drop coating deposition Raman spectroscopy with the goal to verify previously published high sensitivity of the AD spectroscopic diagnosis in cerebral spinal fluid (CSF) and to find a new diagnostic method for blood serum (BS). We optimized measurement conditions for BS. The results were evaluated by the cluster analysis and the principal component analysis. The small set of samples exhibited high sensitivity in both CSF and BS but that distinctly decreased in the whole set. The results for CSF were affected by the choice of the analysed spectral interval. The best for AD diagnose was the interval containing peaks at 980, 1080 and 1249 cm-1.The results for BS have been the most sensitive in the whole spectral range. They have low sensitivity but high specificity for AD (92%). The usage of neural networks has conversely high sensitivity and low specificity in both sets of samples of BS and CSF. Powered by TCPDF (www.tcpdf.org)

<p>U ovom radu je ispitivan uticaj Antitoksičnognutritiva(ATN), preparata na bazi prirodnih aluminosilikata,na biohemijske, fiziolo&scaron;ke, tehnolo&scaron;ke i proizvodne parametre uzgoja pilića. ATN je sme&scaron;a zeolita klinoptilolita, gline monmorilonita i male količine aktivnog uglja. Dodatak ovog preparata u hranivo za piliće u količini od 5 g/kg nije izazvao promene u normalnoj biohemijskoj i fiziolo&scaron;koj homeostazi životinja. Hematolo&scaron;ki parametri, koncentracija metabolita, elektrolita i aktivnost enzima seruma i jetre je bila u granicama referentnih vrednosti. ATN ne utiče na prirast životinja, ni na konverziju hrane, ali dovodi do povećanja relativnih masa pojedinih organa digestivnog trakta.</p><p>Uočava se da dodatkom ATN-a u hranivo dolazi do smanjenja količine masti a povećanja količine proteina u belom mesu. ATN takođe povećava i sadržaj pepela u<br />belom i crvenom mesu.</p><p>Akutni ili hronični tronedeljni oralni unos pojedinih toksikanata (mikotoksina aflatoksina B1i ohratoksina A, herbicida parakvata, jona olova ili toksina<br />cijanobakterija-mikrocistisa) dovodi do poremećaja normalne biohemijske i fiziolo&scaron;ke homeostaze pojedinih organa i tkiva pilića, &scaron;to je utvrđeno na osnovu rezultata hematolo&scaron;kih i biohemijskih analiza, određivanja enzima antioksidativne za&scaron;tite i lipidne&nbsp;<br />peroksidacije, kao i odredjivanja parametara uzgoja i težine organa. ATN, dodat u hranu u količini od 5 g/kg, mogao da bude dobar protektivni agens za delovanje aflatoksina, parakvata, jona te&scaron;kih metala i mikrocistisa, ali ne i ohratoksina.</p> / <p>This study investigated the effects of dietary supplementation with hydrated aluminosilicate (Antitoxic Nutrient-ATN), based on zeolitic ore (clinoptilolite), clay bentonite (montmorillonite),and small amounts of activated charcoal, on&nbsp; performance, hematological, serum, and liver biochemical parameters, as well as organ weights and meat quality in broiler chickens. The dietary addition of ATN has&nbsp; no adverse effects on serum and liver biochemical parameters and does not affect the normal physiological homeostasis of animals. However, the results of this study demonstrate that supplementation with 5 g/kg of ATN influenced organ weights, and chemical composition of broiler chicken meat.</p><p>This study also evaluated the effectiveness of ATN to protect broilers from adverse effects of five different toxic substances (mycotoxins aflatoxin B1, and ochratoxin A, herbicide paraquat, heavy metal ions supplied as lead-acetate, and microcystis, toxin of cyanobacteria). Toxic substances induced oxidative stress and disturb normal biochemical and physiological homeostasis of different tissues and organs in poultry. The results from this study demonstrate that the biochemical variables of serum, liver, kidney, lung and other organs were negatively affected by all five toxic substances. The additionof 5 g/kg of ATN was protective against aflatoxin B1, lead-acetate, paraquat and microcystis, but not against ochratoxin A.</p>

[pt] No presente trabalho foram desenvolvidas duas metodologias analíticas para a determinação de talidomida, um composto de reconhecida importância farmacológica. As metodologias espectrofluorimétrica e voltamétrica desenvolvidas foram comparadas em termos de desempenho analítico.As características fluorescentes e eletroquímicas da talidomida foram estudadas para que se encontrassem condições experimentais que fornecessem máximo sinal fluorescente do analito em solução na temperatura ambiente e possibilidade de pré-concentração do analito no eletrodo de mercúrio.Nas condições experimentais otimizadas para a talidomida, limites de detecção compreendidos entre 10-6 e 10-9 g L-1 foram obtidos para o método espectrofluorimétrico e voltamétrico, respectivamente. Faixas lineares dinâmicas entre 2 e 4 ordens de grandeza foram alcançadas dependendo do método e do interferente presente na amostra. Esses parâmetros de mérito se mostraram adequados para o problema proposto. O possível efeito interferente de substâncias geralmente usadas em associação com o analito foi estudado, e estratégias para minimização das interferências foram desenvolvidas. Enquanto a tetraciclina não interferiu no método espectrofluorimétrico, o uso combinado de meio ácido e irradiação UV da amostra foi necessária para permitir a quantificação do analito em presença de sulfanilamida. Na voltametria, as interferências da tetraciclina e da sulfanilamida puderam ser compensadas pelo uso da quantificação pelo método de adição do analito. Nas determinações dos fluídos biológicos, o uso da extração em coluna de sílica C18 mostrou-se bastante eficiente na separação do analito dos interferentes da matriz para o método espectrofluorimétrico e para o voltamétrico, no caso do soro sangüíneo. Para a urina, apenas a clarificação da amostra com sulfato de amônio foi suficiente no caso da determinação voltamétrica.Os métodos desenvolvidos foram testados na dosagem de talidomida presente em uma formulação comercial e em amostras de urina e soro sangüíneo enriquecidas com o analito. Para tal, utilizaram-se os procedimentos de curva de calibração (espectrofluorimetria) e método da adição do analito (voltametria). Em todos os casos, as recuperações obtidas estiveram compreendidas na faixa de 96,5 a 107,6 %, dentro da faixa de recuperação estabelecida pela Farmacopéia dos Estados Unidos da América. / [en] In the present work two analytical methodologies were developed aiming the determination of thalidomide, an important pharmacological compound. The developed spectrofluorimetric and the voltammetric based analytical methodologies were compared in terms of analytical performance. The thalidomide fluorescent and the electrochemical characteristics were studied in order to find experimental conditions for maximum fluorescence in solution and at room temperature and to allow analyte pre- concentration on the mercury electrode. Using the optimized experimental conditions, limits of detection between 10-6 to 10-9 g L-1 were achieved respectively for the spectrofluorimetric method and for the voltammetric method. Dynamic linear ranges between 2 and 4 orders of magnitude were obtained depending on the method utilized and the interferent substances present in the sample. Those parameters of merit were suitable for this proposed analytical problem. The potential interference effect from substances usually used in association with thalidomide, were studied and strategies for the minimization of such interferences were developed. While no interference in the spectrofluorimetric method was observed for tetracycline, the combined use of acidic medium and UV irradiation of the samples was necessary to allow the analyte determination in the presence of sulfanilamide. For the voltammetric method, interferences from tetracycline and sulfanilamide could be compensated by quantifying thalidomide using the analyte addition method. For the determination in biological fluids, the use a solid-liquid extraction on a C18 column was found to be very effective for the analyte separation and elimination of matrix interferences for the spectrofluorimetric method and for the voltammetric method developed for blood serum. For urine samples, a clean-up step using ammonium sulfate was found to be sufficient for the voltammetric determination of thalidomide. The developed methodologies were tested by determining the thalidomide content in a commercial pharmaceutical formulation and in analyte spiked biological fluids using calibration curves (spectrofluorimetric) and analyte addition method (voltammetric). In all cases, the recoveries were between the 96,5 and 107,6 %, within the recovery range considered adequate according to the United States Pharmacopoeia.

In contrast to mammals, some fish and amphibians have retained the ability to regenerate complex body structures or organs, such as the limb, the tail, the eye lens or even parts of the heart. One major difference in the response to injury is the appearance of a mesenchymal growth zone or blastema in these regenerative species instead of the scarring seen in mammals. This blastema is thought to largely derive from the dedifferentiation of various functional cell types, such as skeletal muscle, skin and cartilage. In the case of multinucleated skeletal muscle fibres, cell cycle re-entry into S-phase as well as fragmentation into mononucleated progenitors is observed both in vitro and in vivo. In order to identify molecules that initiate dedifferentiation of cells at the wound site in amphibians we have established a cellular assay with a cultured newt myogenic cell line. Using this assay we have found a serum activity that stimulates cell cycle re-entry in differentiated multinucleated newt myotubes. The activity is present in serum of all mammalian species tested so far and, interestingly, thrombin proteolysis amplifies the activity from both serum and plasma. We think this serum factor provides a link between wounding and regeneration and its identification will be a key step in understanding the remarkable differences in wound healing between mammals and amphibians. In the course of this PhD thesis we have characterized the serum factor as a thermo-labile, pH- and proteinase K-sensitive, high molecular weight protein that is resistant to denaturing conditions such as SDS, urea or organic solvents. Surprisingly, under denaturing conditions the activity behaves as a low molecular weight protein that displays charge heterogeneity on isoelectric focusing. Using these characteristics of the serum factor we have performed a systematic investigation of commonly used protein chromatography modes and separation techniques to develop a successful purification procedure. After four column chromatography steps -- cation exchange, hydrophobic interaction, heparin affinity and size exclusion chromatography under denaturing conditions -- we have achieved a 2,000-fold purification starting from a commercially available Crude Bovine Thrombin preparation. This represents about 40,000-fold purification over bovine serum. Silver stained gels of the most purified fractions revealed ten major protein bands. In order to finally identify the cell cycle re-entry factor, we are currently analyzing the purification by quantitative mass spectrometry by correlating the abundance of tryptic peptides with activity in sequential fractions across a chromatography run.