The present study investigated the expression profiles of nine genes involved in immune defense of the Pacific white shrimp Litopenaeus vannamei and their responses to oral administration of beta-1,3-glucan. The nine immune related genes were beta-glucan binding protein-high density lipoprotein (BGBP-HDL), lipopolysaccharide/beta-glucan binding protein (LGBP), hemocyanin, prophenoloxidase (proPO), transglutaminase (TGase), penaeidin-3 (PEN-3), crustin, cytosolic manganese superoxide dismutase (cMnSOD), and lysozyme. A series of experiments were carried out in the study including: (1) cDNA cloning and characterization of proPO and LGBP; (2) tissue mRNA expressions of the nine genes in adult shrimp; (3) expression and localization of the nine genes during larval and postlarval ontogenic development; (4) the effects of dietary beta-1,3-glucan on the expression of the nine genes. The cDNA cloning study showed that the proPO cDNA contains an open reading frame of 2061 bp and encodes a 686 amino-acid peptide. The protein sequence of the proPO has a similarity of 85% with those of Penaeus monodon and P. semisulcatus and has an identity of between 58 and 77% with other crustaceans. Northern blot analysis revealed that proPO was constitutively expressed mainly in hemocytes. Its transcripts were observed in hemocytes and many other tissues when detected with RT-PCR. The results of in situ hybridizations showed that the hemocytes that infiltrated in tissues were responsible for the positive signals. The LGBP cDNA contains an open reading frame of 1104 bp and encodes a 367 amino-acid protein with a 17 a. a. signal peptide. The protein sequence of the LGBP has a similarity of 97% with LGBP of L. stylirostris, >90% identity with BGBP of P. monodon and LGBP of Fenneropenaeus chinensis and has an identity of between 63 and 86% with other crustaceans. Northern blot analysis revealed that LGBP was constitutively expressed mainly in hepatopancreas. The results of in situ hybridizations showed that the hepatopancreatic F cells might be the major cell type for LGBP production. Using the complete cDNAs of proPO and LGBP and partial fragments of the other seven genes, their tissue expressions were analyzed by conventional RT-PCR, quantitative real time PCR (qPCR) and in situ hybridization. The results demonstrated that BGBP-HDL, LGBP and hemocyanin were mainly expressed in the hepatopancreas and their expressions levels were about 10 to 30% those of

Orientador: Aulus Cavalieri Carciofi / Banca: Helio Jose Montassier / Banca: Iracilda Zeppone Carlos / Resumo: Vários derivados da parede celular da levedura Saccharomyces cerevisiae conhecidamente agem sobre a imunidade, no entanto a ação, especialmente da fração beta-glucano, foi pouco demonstrada em cães. Para o estudo dos possíveis efeitos sobre a imunidade na espécie canina foram empregadas quatro dietas isonutrientes, contendo uma fonte de parede celular de levedura (PCL), duas fontes de beta-glucano (BG1 e BG2) e uma dieta controle (CT). Foram utilizados 24 cães da raça beagle, adultos, divididos em quatro grupos de seis animais. As dietas foram fornecidas por um período total de 126 dias e as avaliações incluíram hemograma e avaliações bioquímicas, dosagem de anticorpos anti-Leptospira, imunofenotipagem de linfócitos sanguíneos, avaliação da concentração de IgA em fezes, teste de hipersensibilidade cutânea tardia e dosagens de citocinas em sobrenadante de cultura celular. Os animais foram submetidos a desafio antigênico com vacina contra leptospirose no dia 42. Os dados foram avaliados pelo procedimento GLM do SAS, sendo as médias comparadas pelo teste de Tukey (p≤0,1). Nos exames bioquímicos houve discreta variação entre os tratamentos e ao longo dos dias. No hemograma notou-se aumento dos linfócitos para BG2. A dosagem de anticorpos anti-Leptospira, mostrou baixos títulos, não havendo boa resposta à vacinação. A imunofenotipagem revelou um aumento dos linfócitos T totais, T helper, T citotóxicos e linfócitos B no grupo BG2 e de linfócitos T citotóxicos e linfócitos B para o grupo PCL. Apesar da variação da concentração de IgA fecal ao longo dos dias, os tratamentos não influenciaram tais parâmetros. O teste de hipersensibilidade cutânea tardia mostrou um aumento na resposta à inoculação da vacina, para os grupos PCL e BG2. Na dosagem de citocinas em sobrenadante de cultura celular, apenas foi observada diferença na quantificação de TNF-α, ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Some products from the cell wall of the yeast Saccharomyces cerevisiae are known to act on the immunity, however this action, especially of the beta-glucan fraction, has never been demonstrated in dogs. To study these effects on the immunity of dogs four isonutrient diets were made, containing one source of yeast cell wall (YCW), two sources of beta-glucan (BG1 and BG2) and a control diet (CT). 24 adult beagle dogs were used, divided in four groups of six animals. Diets were given for a 126 days period. Evaluations included complete blood count and biochemistry profile, quantification of antibodies against Leptospira, immunophenotyping of blood lymphocytes, IgA concentration in feces, delayed-type hypersensitivity test and quantification of cytokines in cell culture supernatant. Animals were exposed to antigen challenge, by the vaccine against leptospirosis on day 42. Data were analyzed by the GLM procedure of the SAS software and the means were compared by the Tukey test (p<0,01). Biochemistry profile showed slight differences among the groups. A increase in lymphocyte count was observed for BG2 treatment. Quantification of antibodies against Leptospira showed low titles, with poor response to vaccination. Immunophenotyping revealed an increase during the time in total T cells, helper and cytotoxic T cells, and B lymphocytes for BG2 and of cytotoxic T cells and B lymphocytes for YCW group. Despite the variation in fecal IgA concentration during the time, treatments did not influence these parameters. Delayed-type hypersensitivity test showed an increased response to the vaccine inoculation, for YCW and BG2 groups. In the quantification of cytokines in cell culture supernatant the only difference observed was in TNF-α concentration, being BG2 higher than CT. We concluded that both yeast cell wall and beta-glucan fraction act on dogs' immunity / Mestre

Made available in DSpace on 2014-06-11T19:23:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-23Bitstream added on 2014-06-13T20:11:36Z : No. of bitstreams: 1 zaine_l_me_jabo.pdf: 606188 bytes, checksum: da0b30ee696bae955544945c1be515d9 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Biorigin Sa / Vários derivados da parede celular da levedura Saccharomyces cerevisiae conhecidamente agem sobre a imunidade, no entanto a ação, especialmente da fração beta-glucano, foi pouco demonstrada em cães. Para o estudo dos possíveis efeitos sobre a imunidade na espécie canina foram empregadas quatro dietas isonutrientes, contendo uma fonte de parede celular de levedura (PCL), duas fontes de beta-glucano (BG1 e BG2) e uma dieta controle (CT). Foram utilizados 24 cães da raça beagle, adultos, divididos em quatro grupos de seis animais. As dietas foram fornecidas por um período total de 126 dias e as avaliações incluíram hemograma e avaliações bioquímicas, dosagem de anticorpos anti-Leptospira, imunofenotipagem de linfócitos sanguíneos, avaliação da concentração de IgA em fezes, teste de hipersensibilidade cutânea tardia e dosagens de citocinas em sobrenadante de cultura celular. Os animais foram submetidos a desafio antigênico com vacina contra leptospirose no dia 42. Os dados foram avaliados pelo procedimento GLM do SAS, sendo as médias comparadas pelo teste de Tukey (p≤0,1). Nos exames bioquímicos houve discreta variação entre os tratamentos e ao longo dos dias. No hemograma notou-se aumento dos linfócitos para BG2. A dosagem de anticorpos anti-Leptospira, mostrou baixos títulos, não havendo boa resposta à vacinação. A imunofenotipagem revelou um aumento dos linfócitos T totais, T helper, T citotóxicos e linfócitos B no grupo BG2 e de linfócitos T citotóxicos e linfócitos B para o grupo PCL. Apesar da variação da concentração de IgA fecal ao longo dos dias, os tratamentos não influenciaram tais parâmetros. O teste de hipersensibilidade cutânea tardia mostrou um aumento na resposta à inoculação da vacina, para os grupos PCL e BG2. Na dosagem de citocinas em sobrenadante de cultura celular, apenas foi observada diferença na quantificação de TNF-α,... / Some products from the cell wall of the yeast Saccharomyces cerevisiae are known to act on the immunity, however this action, especially of the beta-glucan fraction, has never been demonstrated in dogs. To study these effects on the immunity of dogs four isonutrient diets were made, containing one source of yeast cell wall (YCW), two sources of beta-glucan (BG1 and BG2) and a control diet (CT). 24 adult beagle dogs were used, divided in four groups of six animals. Diets were given for a 126 days period. Evaluations included complete blood count and biochemistry profile, quantification of antibodies against Leptospira, immunophenotyping of blood lymphocytes, IgA concentration in feces, delayed-type hypersensitivity test and quantification of cytokines in cell culture supernatant. Animals were exposed to antigen challenge, by the vaccine against leptospirosis on day 42. Data were analyzed by the GLM procedure of the SAS software and the means were compared by the Tukey test (p<0,01). Biochemistry profile showed slight differences among the groups. A increase in lymphocyte count was observed for BG2 treatment. Quantification of antibodies against Leptospira showed low titles, with poor response to vaccination. Immunophenotyping revealed an increase during the time in total T cells, helper and cytotoxic T cells, and B lymphocytes for BG2 and of cytotoxic T cells and B lymphocytes for YCW group. Despite the variation in fecal IgA concentration during the time, treatments did not influence these parameters. Delayed-type hypersensitivity test showed an increased response to the vaccine inoculation, for YCW and BG2 groups. In the quantification of cytokines in cell culture supernatant the only difference observed was in TNF-α concentration, being BG2 higher than CT. We concluded that both yeast cell wall and beta-glucan fraction act on dogs` immunity

Les maladies cardiovasculaires représentent la principale cause de décès dans le monde et elles sont souvent causées par l’athérosclérose coronarienne caractérisée par l’accumulation de plaques dans la paroi des artères menant à leur rétrécissement et à la réduction ou l’abolition de l’apport sanguin régional au muscle cardiaque. Les anomalies du métabolisme des lipides, incluant l’élévation du cholestérol dans les lipoprotéines de basse densité (LDL-cholestérol) contribuent au processus d’athérosclérose. Plusieurs traitements existent pour réduire les taux de lipides plasmatiques chez les sujets à risque, dont le principal est les statines. Cependant, les statines ne permettent pas toujours d’atteindre les cibles thérapeutiques, ce qui justifie la recherche de nouveaux traitements pouvant modifier les lipides plasmatiques. Une avenue potentielle de traitement à explorer est le beta-glucane, une fibre soluble contenue dans l’avoine pour laquelle certaines données suggèrent un effet hypolipidémiant. L’objectif de ce mémoire était d’évaluer l’efficacité d’un supplément de beta-glucane dans la réduction des taux plasmatiques de LDL-C chez les patients atteints de dyslipidémie à travers le Canada. Pour ce faire, nous avons effectué un essai clinique randomisé en double aveugle contrôlé par placebo dans une population de 264 sujets atteints d’hyperlipidémie qui ont reçus aléatoirement des traitements de beta-glucane à doses de 1.5 g, 3 g et 6 g ou un placebo durant 12 semaines. Tout au long du traitement, des mesures du taux plasmatique de LDL-C ont été effectuées et des analyses statistiques de covariance (ANCOVA) ont été faites pour comparer ces taux entre les groupes actifs et placebos afin d’observer s’il y avait des changements significatifs. La mesure d’efficacité établie était une réduction du taux de LDL-C d’au moins 0.30 mmol/L. Nos résultats suggèrent que le supplément de beta-glucane à doses de 1.5 g, 3 g et 6 g n’est pas efficace pour réduire les taux de lipides sériques. Il n’y a pas eu de variation significative dans les taux de LDL-C lorsque les groupes actifs ont été comparé au groupe placebo. / Cardiovascular diseases are the principal cause of death in the world, and they are often caused by coronary atherosclerosis characterized by the accumulation of plaques in the wall of arteries leading to their narrowing and reduction or disappearance of regional blood supply to the cardiac muscle. Abnormalities of lipid metabolism, including elevated cholesterol in low-density lipoproteins (LDL-C), contribute to the atherosclerotic process. Several treatments exist to reduce levels of plasma lipids in patients at risk, the main one being statins. However, a substantial number of patients do not reach recommended therapeutic targets while treated with statins which creates the impetus for the search of new lipid-modifying treatments. One potential avenue of treatment worthy of investigation is beta-glucan, a soluble fiber found in oats, for which some data have suggested a lipid-lowering effect. The objective of this thesis was to evaluate the efficacy of a beta-glucan supplement in reducing plasma LDL-C levels in patients with dyslipidemia across Canada. To do this, we conducted a randomized, double-blind, placebo-controlled clinical trial in a population of 264 subjects with hyperlipidemia who randomly received beta-glucan treatments at doses of 1.5 g, 3 g and 6 g or a placebo for 12 weeks. Throughout the treatment, LDL-C plasma measurements were taken and statistical analyzes of covariance (ANCOVA) were done to compare these levels between the active and placebo groups to observe if there were any significant changes. The established measure of efficacy was a reduction in LDL-C of at least 0.30 mmol/L. Briefly, our results have shown that beta-glucan in doses of 1.5 g, 3 g and 6 g is not efficacious in reducing plasma lipid levels. There was no significant change in LDL-C levels when the active groups were compared to the placebo group.

No description available.

Fish oil is an excellent source of long chain polyunsaturated fatty acids (LC-PUFA), which can reduce the risk of cardiovascular disease in addition to other health benefits. However, the average intake of LC-PUFA in the Western diet is much lower than the recommended levels. Fish oil is prone to oxidative deterioration when exposed to oxygen and thus must be protected in order to be used in food products. Microencapsulation is one possibility that is already applied by the industry to protect fish oil. However, most of the conventional microencapsulation techniques suffer from shortcomings such as harsh processing conditions or the use of numerous chemicals. The main objective of this thesis was to develop a novel spray process to microencapsulate fish oil based on supercritical fluid (SCF) technology using supercritical carbon dioxide (SC-CO2) and CO2-expanded ethanol (CX EtOH). Fundamental physicochemical properties essential for optimal process design were lacking in the literature; therefore, density, interfacial tension (IFT) and viscosity of fish oil in the form of triglycerides and fatty acid ethyl esters were determined at different temperatures and pressures. Fish oil when equilibrated with SC-CO2 at elevated pressure expanded by up to about 40% in volume and increased in density by up to about 5%. Furthermore, IFT of fish oil in contact with SC-CO2 decreased substantially by an order of magnitude with an increase in CO2 pressure. When fish oil was in contact with CX EtOH, IFT decreased to ultra low levels at pressures of less than 10 MPa. Viscosity of fish oil equilibrated with SC-CO2 decreased substantially with pressure but increased with shear rate. Based on the physicochemical properties determined in this research, a novel process to produce micro- and nano-sized particles containing fish oil was developed based on a SCF spray-drying method. Key processing parameters have been evaluated and can be further optimized to improve encapsulation efficiency. Determination of physicochemical properties contributed to the fundamental understanding of the behavior of the fish oil+CO2 system with and without ethanol under high pressure conditions. The new microencapsulation process shows great potential for the delivery of bioactives in various product applications. / Bioresource and Food Engineering

No description available.

Health claims for barley β-glucan (BG) have prompted the development of more food products using barley. Some new products do not use any form of heat treatment which could become an issue as barley has been found to have high microbial contamination. The aim of this research was to evaluate current commercial barley products for microbial and BG quality and determine the effects of different heat treatments on the safety and physicochemical properties of BG of whole grain barley. Three heat treatments (micronization, roasting and conditioning) were performed on 3 cultivars of barley (CDC Rattan, CDC McGwire and CDC Fibar). The microbial quality was measured with standard plate count (SPC), yeast and mould (MYC), and coliforms/E. coli. Only 4 of the 17 commercial barley products tested met acceptable microbial limits used in this study. All 3 heat treatments reduced SPC, MYC and coliforms to acceptable levels. BG was extracted using an in vitro digestion method to determine its viscosity, molecular weight (MW) and solubility. Heat-treated barley increased the BG viscosity and MW compared to the untreated barley. The effect of heat treatment on starch pasting, particle size and colour were also evaluated. Overall, heat treatments improved the safety and potential health benefits of whole grain barley. / February 2016

The aim of this thesis was to find optimal therapy based on combination of membrane-anchored phagocytic ligands (mannose-(G)5-(K)10-STE, mannan-BAM, mannan-SMCC) with LPS (ligand of signal receptor)for treatment of murine melanoma B16-F10. Mixture of mannan-BAM with LPS applied in pulse regime proved to be the most effective, resulting in heigh reduction of tumor growth and significant prolongation of survival.

Orientadores: Marcia Regina Braga, Marcia Maria Camargo de Morais / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-26T04:21:10Z (GMT). No. of bitstreams: 1 Galvao_DaianeFelbergAntunes_D.pdf: 15313845 bytes, checksum: aa218f685858df886eb7062bfe4337dc (MD5) Previous issue date: 2014 / Resumo: Fruto-oligossacarídeos (FOS) são frutanos de baixo peso molecular produzidos por microorganismos. O interesse em FOS vem aumentando uma vez que eles são considerados ingredientes funcionais benéficos à saúde humana. Com o objetivo de analisar como a produção de FOS e a composição da parede celular de fungos filamentosos é afetada pela fonte de carbono, os fungos Fusarium solani (URM 3338) e Neocosmospora vasinfecta (URM 3329) foram cultivados em meios contendo cinco fontes de carbono diferentes (sacarose, inulina, glucose, frutose ou glucose mais frutose, todos a 1%) e coletas foram realizadas aos 5, 10 e 15 dias de crescimento. A partir do meio de cultivo filtrado foram analisados o pH, teores de açúcar total, açúcares redutores e proteínas, a presença de FOS e atividades enzimáticas invertásica e inulinásica. A partir do micélio, a biomassa foi quantificada e a parede celular foi isolada e sua composição em açúcares neutros, ácidos urônicos e quitina analisada. Foi avaliada também a expressão relativa de genes de síntese de parede celular b-1,3-glucano sintase e quitina sintases. Os dois fungos utilizaram todas as fontes de carbono crescendo nas diferentes condições. Atividade de hidrólise foi detectada no meio contendo sacarose ou inulina para o fungo F. solani, gerando glucose, frutose e fruto-oligossacarideos como produtos havendo utilização dos monossacarídeos. O micélio deste fungo apresentou alterações visíveis no crescimento em meio sólido apenas no meio com frutose, mas foi observada igual quantidade de quitina da parede celular deste fungo quando crescido por cinco dias em sacarose e inulina, mas em menor quantidade com relação aos demais meios. As análises de expressão relativa de genes mostraram indução do gene da b-1,3-glucano sintase e repressão do gene quitina sintase 5 em sacarose e inulina com relação a condição frutose. Estes dados sugerem que a alteração na composição da parede celular do F. solani pode ter relação com a secreção de enzimas nos meios sacarose e inulina. Para N. vasinfecta, quando crescido em sacarose foi observada atividade de transfrutosilação, com a liberação de glucose e síntese de 1-cestose (FOS) no meio. Transfrutosilação também foi observada no meio que teve inulina como fonte de carbono. O micélio deste fungo apresentou alterações visíveis em meio sólido nas condições frutose e inulina, sendo mais hialino do que nas demais condições. A quantidade de quitina na parede celular deste fungo crescido por cinco dias foi maior nas condições frutose e inulina com relação às demais. As análises de expressão relativa de genes mostraram indução dos genes de quitina sintase 4 e 5 nestas duas condições em relação à sacarose. A partir dos resultados, pode-se concluir que as fontes de carbono oferecidas foram utilizadas pelos fungos, que as mesmas afetaram a composição de açúcares da parede celular e a expressão de genes de síntese de componentes da parede e que estes fungos são promissores para a produção de FOS, pois possuem enzimas que hidrolisam a inulina, além de enzimas que sintetizam oligossacarídeos a partir de sacarose por transfrutosilação / Abstract: Fructooligosaccharides (FOS) are low molecular weight fructans produced by microbes and plants. Interest in FOS has been increasing since they are considered as functional food ingredients with benefical effects in human nutrition. With the aim of examining how the production of FOS and the composition of the cell wall of filamentous fungi are affected by the carbon source, Fusarium solani (URM 3338) and Neocosmospora vasinfecta (URM 3329) were cultured in media containing five different carbon sources (sucrose, inulin, glucose, fructose or glucose plus fructose) and samples were taken at 5, 10 and 15 days of growth. From the filtered culture medium, pH, total carbohydrates, reducing sugars and proteins, the presence of FOS and inulinase and invertase activities were analyzed. Mycelium biomass was measured and the cell wall was isolated and its composition in neutral sugars, uronic acids and chitin analyzed. The expression of b-1,3-glucan synthase and chitin synthase genes was also evaluated. Both fungi utilized all the carbon sources for growing. In sucrose- and inulin-containing media, hydrolytic activity was detected in F. solani generating glucose, fructose and FOS as products. When grown on solid culture media, visible changes were observed in mycelium of this fungus only in fructose, but the amount of chitin in the cell wall was higher in the sucrose and inulin-containing media when compared to other carbon sources. The expression b-1,3-glucan synthase gene was induced and chitin synthase 5 gene repressed on sucrose and inulin media. N. vasinfecta showed transfructosilation activity when was grown in sucrose, with release of glucose and synthesis of 1-kestose (FOS) in the culture medium. Transfructosilation was also observed in the inulin-containing medium. The mycelium showed visible changes when the fungus was cultured in solid medium with fructose or inulin as carbon sources. The amount of chitin in the cell wall of this fungus when grown for five days in inulin or fructose was higher in comparison to other carbon sources. The analysis of gene expression showed induction of chitin synthase 4 and 5 genes in these two conditions in relation to sucrose. From the results it can be concluded that the carbon sources affected growth, enzymic activity, composition of the cell wall and gene expression in F. solani and N. vasinfecta, and that these fungi are promising organisms for FOS production since they secrete enzymes that hydrolyze inulin or synthesize oligosaccharides from sucrose by transfructosylation / Doutorado / Biologia Celular / Doutora em Biologia Celular e Estrutural