Typescript. Includes vita and abstract. Bibliography: Includes bibliographical references (leaves 93-101). Description: xi, 101 leaves : ill. ; 29 cm.

Orientador: Valdemiro Carlos Sgarbieri / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-06T19:20:31Z (GMT). No. of bitstreams: 1 Duarte_RenataMariaTeixeira_D.pdf: 1610095 bytes, checksum: f82916ffcf4ea80e18851b0bcf32c734 (MD5) Previous issue date: 2006 / Resumo: Este estudo teve como objetivo o desenvolvimento de um processo de obtenção do plasma bovino desidratado por ¿spray dryer¿ e o estudo dos efeitos de sua adição, comparativamente ao plasma comercial, na dieta de leitões desmamados aos 21 dias de idade, sobre os parâmetros de desempenho (ganho de peso diário (GDP), conversão alimentar (CA) e consumo diário de ração (CDR)), sobre os teores plasmáticos de uréia, proteína total, albumina, globulinas, relação albumina-globulina, além dos níveis de glutationa no fígado e baço e sobre a incidência da diarréia dos animais. Foi estudado também, o efeito da inclusão de plasma na dieta dos animais experimentais na morfologia do intestino delgado dos leitões. O plasma foi obtido a partir do sangue proveniente de bovinos sadios coletado no frigorífico GJota, na cidade de Promissão-SP. Foi utilizado citrato de sódio como anticoagulante e o plasma foi separado da massa celular por centrifugação, congelado e posteriormente desidratado. O plasma foi adicionado à ração dos animais experimentais na proporção de 7% e 2% nas fases I (0-14dias) e II (15-35 dias) do experimento, respectivamente. Durante a primeira fase experimental, os animais alimentados com dietas contendo plasma apresentaram valores de CDR e GDP significativamente superiores (P £ 0,05) aos dos animais da dieta controle (não contendo plasma) e a CA foi melhor para os animais alimentados com o plasma experimental (PE). Não foi encontrada diferença estatisticamente significativa (P > 0,05) para CA entre a dieta contendo plasma comercial (PC) e o controle. Nas três últimas semanas de experimento (Fase II), o GDP foi significativamente superior (P £ 0,05) no grupo de animais que recebeu PE em relação ao controle. Não houve diferença estatística (P > 0,05) para o CDR entre os grupos experimentais. Os animais alimentados com dietas contendo plasma apresentaram melhora na CA, em relação ao controle, de cerca de 10% e 11% para os plasmas comercial e experimental, respectivamente. Considerando o período total de experimento, o plasma não exerceu efeito significativo no CDR (P > 0,05), porém o GDP e a CA foram estatisticamente melhores (P £ 0,05) nos animais alimentados com dietas contendo plasma. O conteúdo de glutationa no fígado dos animais alimentados com proteína de plasma foi significativamente superior (P £ 0,05) aos valores encontrados para o grupo controle, sugerindo que o tipo de proteína presente no concentrado de plasma bovino foi de grande influência no aumento dos níveis de glutationa. Quanto ao estudo da morfologia intestinal, os animais do grupo controle apresentaram valores significativamente inferiores (P £ 0,05) de espessura total da mucosa (ETM), altura de vilosidade (AV) e relação vilosidade:cripta (RVC) na fase I, bem como menor AV na fase II do experimento. A inclusão de plasma na dieta influenciou favoravelmente na redução da incidência da diarréia durante as duas fases experimentais. Conclui-se, portanto, que a inclusão de plasma bovino na dieta de leitões desmamados precocemente atua favoravelmente sobre o desempenho e saúde dos animais / Abstract: The objective of this work was to obtain spray dried bovine plasma (SDBP) and to compare it with a commercial one (CSDBP) as feed ingredients of weanling pigs (weaned at 21 days of age) based on performance, blood and plasma components, liver and spleen glutathione levels, small intestine morphology and diarrhea score. Blood of healthy bovines was collected at a slaughterhouse using sodium citrate as anticoagulant. Blood plasma was obtained by centrifugation and then spray dried. A complete block design with six replications per treatment and four pigs per experimental unit (pen) was used. The treatments consisted on a basal diet and two test diets containing either 7% SDBP or 7% CSDBP in phase I (0-14 day-postweaning period) and 2% SDBP or 2% CSDBP in phase II (15-35 day-postweaning period). In phase I, pigs fed diets containing plasma (either SDBP or CSDBP) had higher (P £ .05) average daily feed intake (ADF) and average daily gain (ADG) than the control pigs. Feed conversion (FC) of pigs fed SDBP was better (P £ .05) then those fed control or CSDBP diets. In phase II, pigs fed SDBP had better ADG and FC (P £ .05) than those fed basal diet. No treatment effects (P > .05) were found on ADF. Although there were no treatment effects (P > .05) on ADF considering the whole experimental period, pigs fed diets containing plasma had greater (P £ .05) ADG and better (P £ .05) FC than those fed basal diet. The liver glutathione levels were higher (P £ .05) for pigs fed plasma diets than for those fed control diet. The animals fed basal diets showed shorter (P £ .05) jejunal mucosa and villus length in phase I, as well as shorter villus length in phase II . The plasma inclusion on the diets also contributed for the reduction on the postweaning diarrhea incidence. Therefore, it can be concluded that bovine plasma provide benefical effects on weanling pig performance and health / Doutorado / Doutor em Alimentos e Nutrição

A fase aguda é caracterizada pela mudança significativa nos níveis de váriasproteínas plasmáticas. As proteínas amilóide sérica A (SAA) e proteína C reativa (PCR), chamadas de proteínas de fase aguda positivas, têm sua concentração plasmática aumentada em até 1000 vezes durante a inflamação. Espera-se que estas proteínas exerçam um papel importante em algumas etapas do processo inflamatório. Para a PCR existem sólidas evidências de sua participação na ativação do sistema complemento, entretanto para a SAA, a função biológica ainda está para ser esclarecida. No plasma, a SAA encontra-se ligada à lipoproteína de densidade alta (HDL), especialmente a fração HDL3. A SAA associa-se à HDL3 através do deslocamento das apolipoproteínas,- preferencialmente a AI e pouco a AlI. Neste trabalho nos propusemos a avaliar a eficiência da passagem de SAA sérica para os focos inflamatórios. Para este fim doseamos, em 20 exsudatos pleurais e 10 exsudatos ascíticos, e respectivos soros, os seguintes parâmetros: proteínas totais (PT), SAA, PCR, apolipoproteína AI, AlI, 8, colesterol total (CT) e triacilgliceróis (TG). As relações exsudato/soro encontrados para estes parâmetros foram: SAA, 0,11 ± 0,14; PCR 0,31 ± 0,20; AI, 0,48 ± 0,26; AlI, 0,63 ± 0,29; 8, 0,46 ± 0,19; PT, 0,67 ± 0,18; CT, 0,34 ± 0,21; TG, 0,36 ± 0,22. Observamos uma forte correlação entre os níveis séricos de PCR versus SAA e entre AI versus All. Para o exsudato foi mantida a correlação forte somente entre AI versus AlI. Nas correlações entre exsudato e soro observamos correlações moderadas apenas para SAA, AlI e PT. Em função dos nossos resultados, concluímos que a proteína de fase aguda SAA tem acesso ao foco inflamatório e que o conteúdo desta proteína em exsudatos é, pelo menos em sua maioria, originário do soro .. A julgar pelos níveis de apo All encontrados no exsudatos, a permeabilidade das membranas que recobrem as cavidades pleurais e ascíticas é maior para HDL3 do que para HDL2 e LDL, fato es e que garante uma passagem eficiente de SAA para as cavidades. A SAA que tem acesso à cavidade deve sofrer extensa proteólise e/ou associar-se eficientemente às células pois a relação de concentrações exsudato/soro encontrada para SAA é menor que para as demais proteínas analisadas. / Inflammation is characterized by profound changes in the serum leveis of acute phase proteins. The huge increase (up to a 1000 fold) in protein serum amyloid A (SAA) and C reactive protein (CRP) suggests that both proteins exert an important role in the inflammatory processo Although it is known the participation of CRP in the activation of the complement complex, the biological role of SAA is not clear yet. In plasma, SAA is associated to the high density lipoprotein (HDL), especially the HDL3 fraction. The association of SAA to HDL3 causes the displacement of the apolipoproteins A-ll and, especially, A-I. The goal of this study was to evaluate the efficiency of the SAA passage from serum to the inflammatory focus. With this purpose we determined total protein (TP), SAA, PC R, A-I, A-ll, apolipoprotein B (B), total cholesterol (TC) and triglicerides (TG) in 20 samples of pleural exudates and 10 samples of ascitic exudates, and respective serum samples. The ratio exudate/serum found were: SAA, 0, 11±0, 14; CRP 0,31±0,20; A-I, 0,48±O,26; A-ll, 0,63±O,29; 8, 0,46±0, 19; TP, 0,67±0,18; TC, 0,34±0,21; TG, 0,36±0,22. There was a high correlation of the serum leveis of CRP versus SAA and AI versus A-ll. A high correlation in the exudate was only found for AI versus All. The correlation between exudate and serum were moderate and were found only for SAA, A-ll and TP. In conclusion we observed that SAA has access to the exudate and that the content of SAA in the exudate is mainly originated from serum. Considering the leveis of A-ll found in the exudates, we supposed that the permeability of the membranes that cover the pleural and ascitic cavities is higher for HDL3 than for HDL2 and LDL. SAA in the inflamed pleural cavity might suffer an extensive proteolysis and/or efficient association to cells because the ratio exudate/serum found for SAA was lower than for other proteins

The biomedical application of nanotechnology has come into the spotlight, with the promise of ‘personalised’ therapeutics that couple early diagnosis with targeted therapeutic activity. Due to the rapid growth of the biomedical applications of nanoparticles, along with the lack of understanding concerning their interactions with biomolecules, there is a pressing need for the development of standard methods directed at investigating the effect of introducing these unique particles into the human body. The central aim of this research is to establish a platform directed at assessing the biological fate of pioneering therapeutic particulate agents, such as metallophthalocyanines (MPcs) and multi-walled carbon nanotubes (FMWCNTs). In particular, we proposed, that Quartz Crystal Microbalance with Dissipation (QCM-D) technology may be employed to assess the composition of blood protein corona deposited on the therapeutic surface, and subsequently assess the biocompatibility of such particles. The proposed method of protein detection utilises the nanogram sensitivity of QCM-D technology to monitor highly specific antibody-antigen interactions. In particular those interactions which occur when probe antibodies are used to detect adsorbed blood proteins deposited on target particle-modified sensor surfaces. Protein detection analysis was directed toward identification of surface bound human serum albumin, complement factor C3c, and human plasma fibrinogen. Preliminary analysis of generic biomedical surfaces indicated human serum albumin demonstrates a higher binding affinity towards positively charged surfaces (i.e. cysteamine self-assembled monolayer), followed by hydrophobic surfaces. Detection of complement C3c, corresponded with literature, where lower levels were detected on negatively charged surfaces (i.e. mercapto undecanoic acid self-assembled monolayer), and higher levels of more hydrophobic surfaces (i.e. 11-amino undecane thiol self-assembled monolayer). Human plasma fibrinogen was observed to favour hydrophilic over hydrophobic self-assembled monolayer surfaces, which was in accordance with literature. Application of the proposed protein detection method for biocompatibility analysis of target therapeutic molecules, namely metallophthalocyanines and acid functionalised multi-walled carbon nanotubes, demonstrated a dependence on modified-surface film characteristics, such as surface charge and topography with regards to human serum albumin and human plasma fibrinogen analysis representing new insights into their potential biomolecular interactions The highest levels of detected human serum albumin and complement C3c were detected on the GePcSmix-modified surfaces. AlPcSmix-modified surfaces analysis suggested the highest levels of human plasma fibrinogen. Two methods of acid functionalisation were employed, using both nitric and sulphuric acid, and pure nitric acid. A general increase in detected human serum albumin, corresponding with an increase in functionalisation time, was observed. Complement C3c detection suggested an increase in deposited complement C3c, with increasing functionalisation time, when assessing nitric acid functionalised multi-walled carbon nanotubes, and a decrease, with increasing functionalisation time, when assessing nitric and sulphuric acid functionalised multi-walled carbon nanotubes. Analysis of human plasma fibrinogen was inconclusive, as were cytotoxicity experiments utilising MCF-7 cells in the presence of metallophthalocyanine complexes, raising simultaneously important considerations for their application and study. In the first such detailed examination of its kind it was concluded that the proposed method of protein detection, using QCM-D, allows for the rudimentary but rapid means of analysis of select protein corona deposited on particulate biomedical surfaces.

M.Sc. / The amount of allozyme variation within, and the extent of genetic differentiation between, 19 sheep breeds from southern Africa were determined by six enzyme coding loci. Another eight enzyme coding loci were analyzed for five breeds. Between 55 and 66.67% of the protein coding loci were polymorphic (95% criterion) in all the breeds, except for the Namaqua sheep that were less polymorphic (33.33%). Values of 1.67 to 2.5 were obtained for the mean number of alleles per locus and average heterozygosities per locus was between 16.6 to 35.9%. The allelic constitution particularly at the transferrin (TF) locus varied appreciably for the different breeds. For example, the TF*H allele was exclusively noted in the Dormer sheep and the TF*G allele was found in the Afiino, Van Rooy, Border Leicester, Blackhead Persian and Skilder-Persian breeds. The only polymorphic breeds at the albumin locus were the South African Mutton Merino and Van Rooy breeds. The allelic constitution at the other polymorphic loci was similar for the breeds, but the allele frequencies of the South African Merino differs from Merino breeds in other countries at the TF locus. Unbiased genetic distance values were the smallest between the Dorper and Dormer breeds and the largest between the Romenof and Blackhead Persian breeds, and the mean genetic distance between the 19 breeds was 0.067. The mean amount of differentiation among the breeds relative to ' the limiting amount under complete fixation (F st) was calculated at 0.123, which is an indication of small genetic differentiation between the breeds studied. However, this , statistic is not reflected by the allele distribution that was not identical for 25 breed pairs (15%) of the total (171) at all the genetic blood systems studied. None of the breed pairs showed identical allele distributions at all the loci studied for at least one locus differed at each breed pair compared. Random amplified polimorphic DNA's gave no consistent or repeatable results. The results of the allozyme study presents the first study of the current genetic characterization of the different southern African sheep breeds.

Resultados obtidos anteriormente pelo nosso grupo mostraram que a proteína de fase aguda amilóide sérica A (SAA) é um potente estímulo para a expressão de mRNA e liberação de TNF-α, IL-1-β e IL-8 em leucócitos humanos, além de atuar como priming para a liberação de espécies reativas de oxigênio (EROs) por neutrófilos. Nosso objetivo, nesse trabalho, foi mostrar a presença de SAA em exsudatos e definir sua origem, além de verificar sua atividade pró-inflamatória in vivo. Para tanto, utilizamos soro e exsudatos pleurais de 32 pacientes com pneumonia. Mostramos primeíramente a presença da SAA no material inflamatório através de SDS-PAGE, immunoblotting e HPLC. A quantificação de SAA nas amostras foi realizada por ELISA. Nestas amostras também foram determinadas as concentrações de proteína total, proteína C reativa (PCR), apo A-I, apo A-II, apo B, colesterol total, triglicérides, TNF-α, IL-1-β e IL-8. A análise integrada dos nossos resultados indica que há uma passagem preferencial da HDL para o foco inflamatório, quando comparada as demais lipoproteínas. A SAA presente em exsudatos é originada do soro e deve sofrer intensa degradação ou associação com células. O efeito da SAA no exsudato é pró-inflamatório, sendo que esta proteína poderia ser um dos alvos para as enzimas proteolíticas e EROs presentes em exsudatos. Acreditamos que este trabalho contribua significamente para a compreensão do, ainda incerto, papel da SAA no processo inflamatório e dá nova abrangência para as funções da HDL e sua participação na reposta imune. / Previous results from our lab showed that the acute phase protein serum amyloid A (SAA) is a potent stimulus for the expression of mRNA and secretion of TNF-α, IL-1-β and IL-8 from human leukocytes. Furthermore SAA primes neutrophils for the generation of reactive oxygen species (ROS). Our goal here was to show the presence of SAA in exudates and define its origin, besides the verification of its proinflammatory activity in vivo. To achieve this goal we used serum and pleural exudates from 32 patients with pneumonia. At first, we showed the presence of SAA in the exudate through SDS-PAGE, immunoblotting and HPLC. SAA was quantified by ELISA. Besides SAA, we also determined the concentrations of total protein, C reactive protein, apo A-I, apo A-II, apo B, cholesterol, triglyceride, TNF-α, IL-1-β e IL-8. The integrate analysis of our results indicates that there is a preferential leakage of HDL to the inflammatory focus when compared to other lipoproteins. SAA present in exudates is originated from serum and may be intensively degraded or associated to cells. The effect of SAA in the exudate is proinflammatory and this protein may be a target for proteolytic enzymes and for ROS present in exudates. We believed that this work adds new insights to the, yet undefined, role of SAA in the inflammatory process and gives a broader compreension to the functions of HDL and its participation in the on the immune response.

In the first part of this thesis, two chromosome Y specific genes ( SRYand TSPY) were chosen as the molecular targets to investigate the characteristics of fetal-specific DNA fragments in maternal plasma. By employing the touch down ligation-mediated PCR coupled with cloning and sequencing, the end property and the fragment species of fetal DNA were studied. / Noninvasive prenatal detection of fetal chromosomal aneuploidies is a much sought-after goal in fetomaternal medicine. The discovery of fetal DNA in the plasma of pregnant women has offered new opportunities for this purpose. However, the fact that fetal DNA amounts to just a minor fraction of all DNA in maternal plasma makes it challenging for locus-specific DNA assays to detect the small increase in sequences derived from a trisomic chromosome. On the other hand, although the clinical applications of plasma DNA for prenatal diagnosis are expanding rapidly, the biological properties of circulating DNA in plasma remain unclear. Recently, next-generation sequencing technologies have transformed the landscape of biomedical research through the ultra-high-throughput sequence information generated in a single run. Massively parallel sequencing allows us to study plasma DNA at an unprecedented resolution and also precisely detect fetal chromosomal aneuploidies in a locus-independent way. / Our group has demonstrated the use of massively parallel sequencing to quantify maternal plasma DNA sequences for the noninvasive prenatal detection of fetal trisomy 21. In the second part of this thesis, the clinical utility of this new sequencing approach was extended to the prenatal detection of fetal trisomy 18 and 13. A region-selection method was developed to minimize the effects of GC content on the diagnostic sensitivity and precision for the prenatal diagnosis of trisomy 13. To facilitate the next-generation sequencing-based maternal plasma DNA analysis for clinical implementation, two measures, i.e., lowering the starting volume of maternal plasma and barcoding multiple maternal plasma samples, were investigated. / Taken together, the results presented in this thesis have demonstrated the clinical utility of massively parallel sequencing of maternal plasma DNA and have also provided us a better understanding of the biology of circulating DNA molecules. / The third part of this thesis focuses on the massively parallel paired-end sequencing of plasma DNA. By analyzing millions of sequenced DNA fragments, the biological properties of maternal plasma DNA were elucidated, such as the size distribution of fetal-derived and maternally-contributed DNA molecules and the potential effect of epigenetic modification on DNA fragmentation. Moreover, the plasma DNA from hematopoietic stem cell transplant patients was characterized by paired-end sequencing approach. These sequencing data not only confirmed the predominant hematopoietic origin of cell-free DNA but also revealed the size difference between hematologically-derived and other tissue-derived DNA molecules in plasma. / Zheng, Wenli. / Adviser: Lo Yu Ming Dennis. / Source: Dissertation Abstracts International, Volume: 73-03, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 261-275). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Includes bibliographical references (leaves 188-217) Insulin-like growth factor-I circulates at high concentrations in blood, mainly complexed with IGF-binding proteins. The main objective of the thesis is to determine the general role played by plasma IGF-binding proteins in the regulation of IGF transfer from blood to tissues.

Diss. Linköping : Linköpings universitet, 2004. / Härtill 7 uppsatser.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2008. / Title from first page of PDF file (viewed Sept. 16, 2008). Includes bibliographical references.