Plant cell wall polysaccharides are abundant natural polymers making them potential sources for sustainable and biodegradable materials. Interfacial behavior, including adsorption and enzymatic degradation, of several plant cell wall polysaccharides and their derivatives were studied with a quartz crystal microbalance with dissipation monitoring (QCM-D), surface plasmon resonance (SPR) and atomic force microscopy (AFM). Xyloglucan adsorption isotherms were obtained to probe how cellulose-hemicellulose interactions were affected by the type of cellulose substrate and molar mass of xyloglucan. Xyloglucan as small as a heptasaccharide still adsorbed irreversibly onto cellulose. Carboxymethyl cellulose (CMC) adsorption onto cellulose and viscoelastic properties and water contents of the adsorbed CMC layers were obtained from a combination of QCM-D and SPR data. The CMC samples formed hydrated and viscoelastic layers compared to the relatively rigid xyloglucan layer. Pectin model surfaces were prepared by pectin adsorption from citric phosphate buffer onto gold substrates. These pectin model surfaces were used for subsequent interaction studies with xyloglucan and enzymatic degradation behavior. There is a strong correlation between the degree of esterification (DE) and film resistance to degradation with the high DE being the most susceptible to degradation. The adsorption of two mixed linkage glucans (MLG), barley and lichen MLG, onto regenerated cellulose (RC) surfaces in the absence and presence of other matrix polysaccharides was studied. Viscoelastic properties of the resulting layer were compared as a function of the proprotion of '-(1''3) linkages with lichen MLG forming softer gel-like layers on RC. The lichen MLG layers were further used for enzymatic degradation studies with respect to enzyme concentration, temperature, pH and ionic strength. These studies show that polymer adsorption is a promising strategy to modify material surfaces and provides fundamental understanding of interactions and biodegradation of cell wall polysaccharides at solid/liquid interfaces. / Ph. D.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The aflatoxin B1 (AFB1) is one of the main mycotoxins that can be identified in foods, and has relevance for agricultural economics and public health because of its immunotoxic properties. A functional immune system is a basic requirement for a healthy life in modern animal production. The interaction involving nutrition and immunity is a strategic factor to obtain a high quality performance in the poultry industry. Immunomodulators such as β-glucans have an immunostimulating activity, which enables the host ability to resist opportunistic infections. To contribute to the elucidation of the mechanism of action of β-glucans in broiler chicken lymphocytes, the effects of the concentrations of 0.1, 1 and 10% of β-glucans derived from Saccharomyces cerevisiae were investigated in lymphocytes exposed to increasing concentrations of AFB1 (0, 0.1, 1, 10, 20 μg/ml). Lymphocytes were separated by Ficoll-Histopaque density and cultured in 96 well-plates containing AFB1 and/or β-glucans in a 5% CO2 atmosphere at 39°C. MTT, PicoGreen® and 2'-7'-dichlorofluorescein diacetate cytotoxicity tests were evaluated at 24, 48 and 72 h of incubation. The comet assay to elucidate the DNA damage was also performed. The percentage of viable cells decreased in the presence of 10 μg/ml AFB1 at 48 h (p < 0.05) and 10 and 20 μg/ml AFB1 at 72 h (p < 0.001 and p < 0.01, respectively), when compared to the control group (0 μg/ml). Furthermore, an increase in cell-free DNA in AFB1 concentrations > 1 μg/ml (p < 0.001) and the generation of ROS at 24 h were also observed. DNA damage increased approximately 2.3 fold in lymphocytes exposed to 20 μg/ml of AFB1 when compared to the control group. Conversely, β-glucans showed cytoprotective effects (p < 0.001), and the concentration of 1% reverted the AFB1-induced lymphocyte damage. β-glucans at 10% significantly increased (p < 0.001) the formation of reactive oxygen species (ROS), potentiating the AFB1-induced ROS formation. In conclusion, this study showed that AFB1 and β-glucans exert influence on lymphocyte oxidative metabolism and have dose-dependent potentiating effects. The results also showed genoprotective in vitro effect of β-glucans in poultry lymphocytes exposed to AFB1, being the concentration of 1% β-glucans able to maintain DNA integrity. / A aflatoxina B1 (AFB1) é uma das principais micotoxinas que podem ser identificadas em alimentos, possuindo relevância agroeconômica e para a saúde pública, por ser considerada imunotóxica. Um sistema imune funcional é um requisito básico para uma vida saudável na produção moderna de animais. A interação envolvendo nutrição e imunidade é um fator estratégico para obter um bom desempenho em frangos de corte. Devido as suas atividades imunomodulatórias, substâncias como as β-glucanas proporcionam ao hospedeiro uma maior capacidade de resistir a infecções oportunistas. Para melhor compreender o mecanismo de ação das β-glucanas, sobre os linfócitos de frangos de corte, investigou-se os efeitos das concentrações de 0.1, 1 e 10% de β-glucanas derivadas do fungo Saccharomyces cerevisiae em linfócitos expostos a crescentes concentrações de AFB1 (0, 0.1, 1, 10, 20 μg/ml). Os linfócitos foram separados através do reagente de densidade Ficoll-Histopaque e cultivados em placas de 96 poços, contendo as concentrações de AFB1 e/ou β-glucanas em atmosfera de 5% de CO2 a 39°C durante 24, 48 e 72 h. A citotoxicidade celular foi avaliada através dos testes MTT e PicoGreen®, e a formação de espécies reativas de oxigênio (EROS) através do ensaio 2′-7′- diacetato diclorofluoresceína. Também foi utilizado o teste cometa para elucidar os danos ao DNA. A viabilidade celular reduziu na presença de 10 μg/ml de AFB1 em 48 h (p < 0.05) e em 10 and 20 μg/ml de AFB1 (p < 0.01 e p < 0.001, respectivamente) em 72 h quando comparada ao grupo controle. Além disso, as concentrações de AFB1 > 1 μg/ml aumentaram significativamente (p < 0.001) a liberação de fita dupla de DNA (dsDNA) e a produção de EROS em 24 h. Os danos causados ao DNA foram confirmados através do momento cauda do cometa e aumentaram cerca de 2,3 vezes em linfócitos expostos a 20 μg/ml de AFB1 quando comparados ao grupo controle. Por outro lado, as β-glucanas exerceram efeitos citoprotetores (p < 0.001), sendo que a concentração de 1% foi capaz de reverter os danos genotóxicos causados pela ação da AFB1. Já a concentração de 10% de β-glucanas aumentou significativamente (p < 0.001) a formação de EROS, potencializando a ação da AFB1. Em conclusão, este estudou evidenciou que a AFB1 e as β-glucanas exercem influência sobre o metabolismo oxidativo dos linfócitos e possui efeito potencializador dose-dependente. Os resultados também evidenciaram o efeito genoprotetor in vitro de β-glucanas em linfócitos de frangos expostos a AFB1, sendo a concentração de β-glucanas a 1% capaz de manter a integridade do DNA.

The content of beta-glucans and arabinoxylans in barley grain, malt and wort was monitored. The varieties of barley were Pionier, Libuše, Manta, Tango, KWS Amadora, Kampa, KWS Irina, Francin, Odyssey, Overture, Vendela, Petrus, Laudis 550, Sunshine, Kangoo, Xanadu, Sebastian, Bojos and Malz. These varieties were grown on three different habitats (Uherský Ostroh, Věrovany and Vysoká u Příbramě). The content of beta-glucans and arabinoxylans was also analysed in five different types of beer. In brewing and malting industry the content of beta-glucans and arabinoxylans is mainly monitored because of their negative influence on decanting of wort and filtration of beer. These substances can contribute to turbidity and precipitation in beer and thus negatively effect the stability of beer. The content of beta-glucans in barley grain, malt, wort and beer was determined by FIA method and the content of arabinoxylans was determined in barley grain, wort and beer by Douglas method. The highest content of beta-glucans in barley grain was found in 14/19 varieties at Uherský Ostroh, where the amount of beta-glucan content was in the range of 3,65-5,60 %. The highest beta-glucan content in malt was found in 17/19 varieties at Vysoká u Příbramě, where the beta-glucan content was between 0,15-0,79 % and the highest content of beta-glucans in wort was found in 16/19 varieties also at Vysoká u Příbramě. There was the beta-content in the range of 22-184 mg/l. The results of content of arabinoxylans in barley grain and wort were more balanced. The highest content of arabinoxylans in barley grain was found in 10/19 varieties at Věrovany. The content of arabinoxylans at this habitat was in the range of 3,04-4,56 %. The highest content of arabinoxylans in wort was determined in 8/19 varieties at Uherský Ostroh. The content of arabinoxylans at this habitat was in the range of 542-1040 mg/l. The highest content of beta-glucans in samples of beer was determined in a sample of light lager (207 mg/l) and the highest content of arabinoxylans was found in a samle of wheat lager (1465 mg/l).

<p dir="ltr">Dietary fibers have been observed to modulate the gut microbiome in ways that prevent and moderate human diseases and confer health benefits onto their human host. How dietary fibers do this is through their structure; gut microbes are equipped with a variety of differ- ent carbohydrate-active enzymes (CAZymes) that allow some to hydrolyze glycosidic bonds, thereby utilizing the dietary fiber. The more complex the dietary fiber, the more diverse the maintained gut microbiota may be, as specialist species may be required for complete hydrol- ysis. Therefore, increasing structural complexity of dietary fibers may increase gut microbial diversity and help prevent diseases. To understand if structural features impact the gut mi- crobiome, a set of resistant glucans varying in structures, including mixed-linkage -glucans, resistant maltodextrins (similar to type IV resistant starch) and polydextroses, which are comprised entirely of glucose, were used as substrates in an in vitro sequential batch fermen- tation using fecal microbiota form three healthy donors as inocula. I measured metabolic outputs, growth curves, and community structures by 16S rRNA amplicon sequencing, which I analyzed for through alpha and beta diversity differences and taxa that overrepresented and increased in each treatment. My results show that, depending on the donor and the resistant glucan, structure does significantly impact the concentrations of short-chain fatty acids (SCFAs) and other metabolites that are produced. Resistant glucan structure also impacts alpha and beta diversity to a degree and linear discriminant analysis (by LEfSe) results also support that specific species have preference towards substrates as well. Next, resistant glucans were supplemented into a high-fat diet, and compared these diets to a low- fat diet (LFD), high-fat diet with cellulose (HFD), and high-fat without cellulose (HWC) in a mouse study using C57BL/6J mice over 4 weeks. Increasing microbial diversity will not only increase diversity in the gut microbiome, but it will also provide protective effects in behavior such as helping to prevent anxiety. I measured weight, metabolic outputs, 16S community structure, changes in alpha and beta diversity, and differential abundances of OTUs and taxa by discriminant analysis Effect Size (LEfSe) and Metastats, and anxiety behaviors using open field and light/dark box tests. Microbial community structure was significantly different in treatment groups from controls. Anxiety for mice in tapioca dextrin 01 (TD01), tapioca dextrin 03 (TD03), and resistant maltodextrin (RMF) treatment groups were gen- erally increased, suggesting that the chemical structure of these resistant dextrins may alter the gut microbiome in ways that may influence behavior.</p><p dir="ltr">My overall results support the hypothesis that the fine structural features of dietary fibers do significantly impact the gut microbiome by selecting for specific microbiota, and may even impact cognition and behavior.</p>

Thesis (MSc (Genetics. Institute of Plant Biotechnology))--Stellenbosch University, 2008. / Traditional methods used for the discovery of novel genes have previously relied upon the ability to culture the relevant microbes and then demonstrate the activity of a specific enzyme. Although these methods have proved successful in the past, they severely limit our access to the genomes of organisms which are not able to be cultured under laboratory conditions. It was therefore the aim of this project to use metagenomic strategies for the identification of novel polymer-producing genes with the prospect of commercial exploitation. In this study, soil-derived metagenomic libraries were functionally screened for potential -glucan producing clones using aniline blue staining. Positive reacting clones were selected and sequenced. Initial sequencing revealed a gene with high homology to previously described glucan synthases, the products of these genes all having significant industrial value. The clone was transformed into a suitable bacterial host, cultured and allowed to produce the polymer of interest. The polysaccharide was purified and subjected to various chemical analyses so as to confirm its monosaccharide composition. Data suggests that this polymer is composed mainly of glucose units and that it may be secreted out of the cell. Purification of the active enzyme was attempted using classical protein purification methods with faint activity being detected using Native polyacrylamide gel electrophoresis (PAGE). Further attempts to demonstrate activity were made through the construction of a GST (glutathione S-transferase) tagged fusion protein. The second part of this study focuses on the construction and screening of a metagenomic DNA library from whey, a by-product of the cheese manufacturing process. It was envisaged that this could provide a resource for the identification of high value polymers when lactose is provided as a sole carbon source. The library was screened for function using Congo Red for the detection of extra-cellular polysaccharides.

Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Os cogumelos há muitos anos vêm sendo utilizados na alimentação por seus valores nutricionais e/ou como agente terapêutico. Um dos fungos mais estudados atualmente é o Agaricus blazei Murrill, popularmente conhecido como Cogumelo do Sol, em função dos efeitos benéficos que lhe é creditado na prevenção de câncer. O estudo analisou os efeitos da ingestão do A. blazei, no parênquima pulmonar de ratos Wistar submetidos a estímulo carcinógeno induzido pela nitrosamina 4-(metilnitrosamina)-1-(3-piridil)-1 butanona NNK, potente carcinogênico pulmonar sistêmico em roedores. Foi preparado extrato aquoso de A. blazei nas doses de 14,3 mg e 42,9 mg administrado por gavagem durante 22 semanas juntamente com a NNK administrada via subcutânea na dose de 1,5 mg de NNK/Kg de peso corporal do animal durante 20 semanas. Foram formados seis grupos experimentais: Grupo Controle (GC); Grupo Cogumelo do Sol nas doses de 14,3 mg e 42,9 mg por gavagem e injeção de óleo de milho subcutânea (CSOL 14,3 e CSOL 42,9); Grupo Nitrosamina (GN) com injeção de NNK subcutânea e salina por gavagem; Grupo Nitrosamina com Cogumelo do Sol nas doses de 14,3 e 42,9 mg (NSOL 14,3 e NSOL 42,9). Os animais foram submetidos à tomografia computadorizada (TC) para monitoramento do surgimento de possíveis tumores, foi analisado também peso corporal, consumo de ração ao longo de todo o experimento e realizada a composição centesimal do A. blazei Murrill. Ao final do experimento os animais foram mortos e foram analisados: hemograma completo, hepatograma, colesterol, triglicerídeo, análise macroscópica e microscópica do pulmão e a análise de curtose da curva do histograma fornecido pelo software BEBÚI utilizado no estudo através da imagem pulmonar da TC. Como conclusão foi verificado que a substância NNK na dose utilizada neste experimento não foi capaz de desenvolver uma neoplasia pulmonar nos animais estudados no período do experimento; no presente modelo experimental as diferenças encontradas nas análises histopatológicas e na análise da curtose mostram que a dose de cogumelo do sol de 14,3 mg se mostrou mais eficaz contra o surgimento de inflamação e opacidades pulmonares se comparada à dose de 42,9 mg. / The mushrooms have been used for many years as food for its nutritional value and/or as a therapeutic agent. One of the most studied fungus is currently the Agaricus blazei Murrill, popularly Known as Cogumelo do Sol, for its beneficial credited in the prevention of cancer. This study examined the effects of ingestion of A. blazei in the lung parenchyma of wistar rats subjected to stimulus-induced carcinogen 4 - (metilnitrosamina) -1 - (3-pyridyl) -1 butanone NNK, a potent systemic lung carcinogen in rodents. Was prepared aqueous extract of A. blazei in doses of 14,3 mg and 42,9 mg administered by gavage during 22 weeks together with NNK administered subcutaneously at a dose of 1,5 mg NNK/Kg body weight of the animal during 20 weeks. Were formed six groups: Control Group (CG); Mushroom Group at doses of 14,3 mg and 42,9 mg by gavage and injection of corn oil subcutaneously (CSOL 14.3 and 42.9); Group Nitrosamine (GN) with subcutaneous injection of NNK and saline by gavage; Group Nitrosamine + Mushroom at doses of 14,3 mg and 42,9 mg (NSOL 14.3 and NSOL 42.9). The animals were submitted to Computed Tomografy (CT) to monitor the possibility of development of lung tumors. Was also examined body weight and food intake throughout the experiment. At the end of 22 weeks, the experiment lasted for over four months for observation of development or growth of tumors via CT. After this period the animals were killed and analyzed: complete blood count, hepatogram, cholesterol, triglycerides, macroscopic and microscopic analysis of lung and kurtosis of the histogram curve supplied by the software used in the study called BEBÚI through the lung CT image. In conclusion it was found that NNK dose used on this study was not able to development a lung cancer in animals studied during the experiment. On this experimental model the differences found in histological analysis and the analysis of kurtosis shows that the dose of A. blazei of 14,3 mg was more effective against inflammation and the appearance of opacities compared to the dose of 42,9 mg.

Made available in DSpace on 2014-12-17T14:03:37Z (GMT). No. of bitstreams: 1 MariliaSNS_TESE.pdf: 2303324 bytes, checksum: d055f1a32ee94b3ffe5f389dbf9dffe0 (MD5) Previous issue date: 2014-04-08 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Compounds derived from fungi has been the subject of many studies in order to broaden the knowledge of their bioactive potential. Polysaccharides from Caripia montagnei have been described to possess anti-inflammatory and antioxidant properties. In this study, glucans extracted from Caripia montagnei mushroom were chemically characterized and their effects evaluated at different doses and intervals of treatment. It was also described their action on colonic injury in the model of colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its action on cells of the human colon carcinoma (HT-29). Compounds extracted of C. montagnei contain high level of carbohydrates (96%), low content of phenolic compounds (1.5%) and low contamination with proteins (2.5%). The (FT-IR) and (NMR) analysis showed that polysaccharides from this species of mushroom are composed of &#945;- and &#946;-glucans. The colonic damage was evaluated by macroscopic, histological, biochemical and immunologic analyses. The results showed a reduction of colonic lesions in all groups treated with the glucans of Caripia montagnei (GCM). GCM significantly reduced the levels of IL-6 (50 and 75 mg/kg, p < 0.05), a major inflammatory cytokine. Biochemical analyses showed that such glucans acted on reducing levels of alkaline phosphatase (75 mg/kg, p < 0.01), nitric oxide (p < 0.001), and myeloperoxidase (p < 0.001). These results were confirmed microscopically by the reduction of cellular infiltration. The increase of catalase activity suggest a protective effect of GCM on colonic tissue, confirming their anti-inflammatory potential. GCM displayed cytostatic activity against HT-29 cells, causing accumulation of cells in G1 phase, blocking the cycle cell progression. Those glucans also showed ability to modulate the adhesion of HT-29 cells to Matrigel? and reduced the oxidative stress. The antiproliferative activity against HT-29 cells displayed by GCM (p <0.001) can be attributed to its cytostatic activity and induction of apoptosis by GCM / Compostos derivados de fungos tem sido alvo de muitos estudos a fim de desenvolver o conhecimento acerca de seu potencial bioativo. Polissacar?deos de Caripia montagnei j? foram descritos por possu?rem propriedades anti-inflamat?ria e antioxidante. Neste estudo, os polissacar?deos extra?dos do fungo Caripia montagnei foram caracterizados quimicamente e seus efeitos sobre as les?es intestinais foram avaliados em diferentes intervalos de tratamento no modelo de colite induzida por ?cido 2,4,6 - trinitrobenzenossulf?nico (TNBS), verificou-se ainda sua a??o sobre c?lulas do carcinoma de c?lon humano, HT-29. Na an?lise realizada no extrato obtido de C. montagnei foi verificado que este ? formado principalmente, por carboidratos (96%) apresentando um baixo teor de compostos fen?licos (1,5%) e baixa contamina??o prot?ica (2,5%). As an?lises por espectroscopia de infra vermelho (FT-IR) e resson?ncia magn?tica nuclear (RMN) mostraram que os polissacar?deos desta esp?cie de fungo s?o &#945; e &#946; -glucanas. O dano col?nico foi avaliado por an?lises macrosc?picas, histol?gicas, bioqu?micas e imunol?gicas. Os resultados mostraram a redu??o das les?es no c?lon em todos os grupos tratados com as glucanas (GCM). GCM reduziram significativamente os n?veis de IL-6 (50 e 75 mg/Kg, p < 0,05), uma importante citocina inflamat?ria. As an?lises bioqu?micas mostraram que essas glucanas atuaram na redu??o dos n?veis de fosfatase alcalina (75 mg/Kg, p < 0,01), ?xido n?trico (p < 0,001) e mieloperoxidase (p < 0,001). Estes resultados foram confirmados pela redu??o da infiltra??o celular observado microscopicamente. O aumento da atividade da catalase, sugere um efeito protetor de GCM no tecido do c?lon, o que confirma o seu potencial anti-inflamat?rio. GCM mostraram atividade citost?tica sobre as c?lulas HT-29, causando ac?mulo de c?lulas na fase G1 e impedindo, assim, a progress?o do ciclo celular. As glucanas deste estudo tamb?m mostraram habilidade em modular a ades?o de c?lulas HT-29 ao Matrigel? e reduzir o estresse oxidativo nessas c?lulas. A atividade antiproliferativa contra c?lulas HT-29 exibida por GCM pode ser atribu?da ? sua a??o citost?tica ou indu??o da apoptose por essas glucanas

Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Os cogumelos há muitos anos vêm sendo utilizados na alimentação por seus valores nutricionais e/ou como agente terapêutico. Um dos fungos mais estudados atualmente é o Agaricus blazei Murrill, popularmente conhecido como Cogumelo do Sol, em função dos efeitos benéficos que lhe é creditado na prevenção de câncer. O estudo analisou os efeitos da ingestão do A. blazei, no parênquima pulmonar de ratos Wistar submetidos a estímulo carcinógeno induzido pela nitrosamina 4-(metilnitrosamina)-1-(3-piridil)-1 butanona NNK, potente carcinogênico pulmonar sistêmico em roedores. Foi preparado extrato aquoso de A. blazei nas doses de 14,3 mg e 42,9 mg administrado por gavagem durante 22 semanas juntamente com a NNK administrada via subcutânea na dose de 1,5 mg de NNK/Kg de peso corporal do animal durante 20 semanas. Foram formados seis grupos experimentais: Grupo Controle (GC); Grupo Cogumelo do Sol nas doses de 14,3 mg e 42,9 mg por gavagem e injeção de óleo de milho subcutânea (CSOL 14,3 e CSOL 42,9); Grupo Nitrosamina (GN) com injeção de NNK subcutânea e salina por gavagem; Grupo Nitrosamina com Cogumelo do Sol nas doses de 14,3 e 42,9 mg (NSOL 14,3 e NSOL 42,9). Os animais foram submetidos à tomografia computadorizada (TC) para monitoramento do surgimento de possíveis tumores, foi analisado também peso corporal, consumo de ração ao longo de todo o experimento e realizada a composição centesimal do A. blazei Murrill. Ao final do experimento os animais foram mortos e foram analisados: hemograma completo, hepatograma, colesterol, triglicerídeo, análise macroscópica e microscópica do pulmão e a análise de curtose da curva do histograma fornecido pelo software BEBÚI utilizado no estudo através da imagem pulmonar da TC. Como conclusão foi verificado que a substância NNK na dose utilizada neste experimento não foi capaz de desenvolver uma neoplasia pulmonar nos animais estudados no período do experimento; no presente modelo experimental as diferenças encontradas nas análises histopatológicas e na análise da curtose mostram que a dose de cogumelo do sol de 14,3 mg se mostrou mais eficaz contra o surgimento de inflamação e opacidades pulmonares se comparada à dose de 42,9 mg. / The mushrooms have been used for many years as food for its nutritional value and/or as a therapeutic agent. One of the most studied fungus is currently the Agaricus blazei Murrill, popularly Known as Cogumelo do Sol, for its beneficial credited in the prevention of cancer. This study examined the effects of ingestion of A. blazei in the lung parenchyma of wistar rats subjected to stimulus-induced carcinogen 4 - (metilnitrosamina) -1 - (3-pyridyl) -1 butanone NNK, a potent systemic lung carcinogen in rodents. Was prepared aqueous extract of A. blazei in doses of 14,3 mg and 42,9 mg administered by gavage during 22 weeks together with NNK administered subcutaneously at a dose of 1,5 mg NNK/Kg body weight of the animal during 20 weeks. Were formed six groups: Control Group (CG); Mushroom Group at doses of 14,3 mg and 42,9 mg by gavage and injection of corn oil subcutaneously (CSOL 14.3 and 42.9); Group Nitrosamine (GN) with subcutaneous injection of NNK and saline by gavage; Group Nitrosamine + Mushroom at doses of 14,3 mg and 42,9 mg (NSOL 14.3 and NSOL 42.9). The animals were submitted to Computed Tomografy (CT) to monitor the possibility of development of lung tumors. Was also examined body weight and food intake throughout the experiment. At the end of 22 weeks, the experiment lasted for over four months for observation of development or growth of tumors via CT. After this period the animals were killed and analyzed: complete blood count, hepatogram, cholesterol, triglycerides, macroscopic and microscopic analysis of lung and kurtosis of the histogram curve supplied by the software used in the study called BEBÚI through the lung CT image. In conclusion it was found that NNK dose used on this study was not able to development a lung cancer in animals studied during the experiment. On this experimental model the differences found in histological analysis and the analysis of kurtosis shows that the dose of A. blazei of 14,3 mg was more effective against inflammation and the appearance of opacities compared to the dose of 42,9 mg.

Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-10-20T17:56:33Z No. of bitstreams: 2 Dissertação - Estefânia Viano da Silva - 2014.pdf: 4645592 bytes, checksum: f3321dacb1ccfd2f2b017d08f8e3b578 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-10-20T17:58:09Z (GMT) No. of bitstreams: 2 Dissertação - Estefânia Viano da Silva - 2014.pdf: 4645592 bytes, checksum: f3321dacb1ccfd2f2b017d08f8e3b578 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-10-20T17:58:09Z (GMT). No. of bitstreams: 2 Dissertação - Estefânia Viano da Silva - 2014.pdf: 4645592 bytes, checksum: f3321dacb1ccfd2f2b017d08f8e3b578 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-06-26 / The mushrooms have been used as food and medicine for thousands of years. Among them, Grifola frondosa (“Maitake”) may be on the most versatile and promising medicinal mushrooms for use as a dietary supplement. Besides the high nutritional value, they are a good source of compounds such as carbohydrates that can act as biological response modifiers. In this work, β-D-glucans (PSF2-GfM and MRSF2K-GfM fractions) and heteropolysaccharides unusual in the literature (MEPF3CW-GfM and PF2K-GfM fractions) were isolated from the mycelium, grown in solid culture, of the G. frondosa, by successive aqueous and alkaline extractions, followed by fractionation through freeze-thawing, precipitation of soluble material with a Fehling solution, and dialysis using a membrane with an exclusion limit of 1000 kDa. The chemical structures were established based on monossacharide composition, periodate oxidation, methylation analysis, HPSEC-MALLS and NMR studies. The branched β-D-glucans (GLC-GfM) found were similar to those previously described for the fungus, which contains a (13)- linked β-Glcp main-chain, substituted at O-6 by single-unit -Glcp side-chains, on the average of one to every third residues of the backbone, but with different molar masses and water solubility. In adittion, novel heteropolymers were obtained. One was a fucomannogalactan (FMG-GfM), with a molar mass of 175 x 103 g mol-1, was shown to have a main chain of (1→6)-linked α-D-Galp and 3-O-Me-α-DGalp, both of which are partially substituted at O-2, principally, by 3-O-α-D-Manp-α-LFucp groups. The other was a fucoxylomannan (FXM-GfM) (Mw 202 x 103 g.mol-1), which consisted of backbone of (13)-linked α-D-Manp units, totally substituted at O-4 mainly by 4-O-α-L-Fucp-β-D-Xylp disaccharide. Thus, the polymers obtained from Grifola frondosa may could be good candidates for evaluation of its biological effect, considering the results obtained for others similar mushrooms polysaccharides. / Os cogumelos têm sido usados como alimentos e remédios por milhares de anos. Entre eles, o “Maitake” (Grifola frondosa) pode ser um dos cogumelos medicinais mais versáteis e promissores para o uso como um suplemento dietético. Além do valor nutricional elevado, eles são uma boa fonte de compostos, como os carboidratos, que podem agir como modificadores de resposta biológica. Neste trabalho, foram isolados do micélio (cultivado em meio sólido) de G. frondosa as β-D-glucanas (frações PSF2KGfM e MRSF2K-GfM) e heteropolissacarídeos (frações MePF3CW-GfM e PF2K-GfM) não comumente encontrados na literatura, por sucessivas extrações aquosas e alcalinas, seguido por fracionamento através do processo de congelamento/degelo, precipitação das frações solúveis em água com solução de Fehling e diálise em membranas com limite de exclusão de 1000 kDa. A estrutura química foi determinada através das análises de composição mossacarídica, oxidação com periodato de sódio, metilação, HPSEC-MALLS e RMN. As β-D-glucanas (GLC) isoladas foram similares àquelas previamente descritas para este fungo, as quais contém uma cadeia principal formada por unidades de 3-O-β-D-Glcp, podendo ser substituídas em O-6 por terminais não redutores de β-Glcp (1 ramificação a cada três unidades da cadeia principal), distinguindo-as quanto à solubilidade em água e tamanho (massa molar). Além destas, foram obtidos novos heteropolímeros. Um deles foi uma fucomanogalactana (FMGGfM), com uma massa molar de 175 x 103 g mol-1, a qual possui um core constituído por unidades de 6-O-α-D-Galp e 6-O-(3-O-Me-α-D-Galp), sendo ambas parcialmente substituídas em O-2 por 3-O-α-D-Manp-α-L-Fucp. O outro heteropolissacarídeo isolado consiste em uma fucoxilomanana (FXM-GfM) (Mw 202 x 103 g.mol-1) formada por unidades de α-D-Manp unidas por ligações glicosídicas do tipo (13) as quais se encontram ramificadas em O-4 principalmente pelo dissacarídeo 4-O-α-L-Fucp-β-DXylp. Considerando os efeitos terapêuticos apresentados por moléculas similares descritas para macrofungos, estes podem ser bons candidatos para avaliação do efeito biológico.

En la presente Tesis Doctoral se utilizaron cebada y remolacha como fuentes alternativas de carbohidratos en dietas para trucha arcoíris con el objetivo de evaluar su efecto sobre parámetros productivos, histología hepática e intestinal, estrés y calidad de la carne. Los marcadores estudiados fueron los mismo tanto para los experimentos de cebada como de remolacha. En las pruebas de la cebada se utilizaron concentraciones crecientes de este ingrediente (0-32%) en la dieta, se inició con un peso medio de 127.72 ± 5.65 g y se finalizó cuando alcanzaron el peso comercial al cabo de 84 días. En las pruebas de remolacha se utilizaron dos niveles de remolacha (14% y 28%) y dos de betaína (0.9% y 1.63%), se inició con un peso medio de 69 ± 2.2 g y se finalizó cuando los peces alcanzaron el peso comercial al cabo de 105 días. La inclusión de cebada en dietas de trucha arcoíris no mostró diferencias significativas en los parámetros de crecimiento ni biométricos. La inclusión de 14% remolacha y 0.9% betaína no afectó al crecimiento, parámetros nutritivos, biométricos y retenciones de nutrientes en comparación con el control, concentraciones mayores de remolacha y betaína tuvieron un efecto negativo. Cuando se estudiaron los parámetros histológicos y morfométricos en hígado e intestino, los resultados mostraron que los peces alimentados con concentraciones crecientes de cebada mostraron hepatocitos más pequeños, mientras que los peces alimentados con remolacha y betaína presentaban hepatocitos más grandes, en ambos casos el hígado mostró un menor nivel de vacuolización. La concentración de cebada de un 8% produjo un efecto negativo a nivel morfológico del intestino, sin embargo, la inclusión de remolacha no produjo ningún efecto. Los peces fueron sometidos a estrés por ausencia de oxígeno y aumento de densidad durante 10 minutos. Cuando el estrés fue analizado a nivel bioquímico los resultados mostraron que a niveles basales, la inclusión de cebada y remolacha no causó ningún cambio en los niveles de cortisol, glucosa y MDA, aunque los valores basales de lactato fueron significativamente más bajos en los peces alimentados con remolacha y betaína. Treinta minutos después del estrés la concentración de cortisol, glucosa y lactato aumentó significativamente en los peces de todos los grupos y la concentración de MDA disminuyó. La inclusión de cebada mostró valores más bajos de cortisol, glucosa y lactato que la dieta control. Cuando los peces alcanzaron el peso comercial se analizó la calidad de los filetes y las propiedades antioxidantes. En ambas pruebas los peces alimentados con los ingredientes objeto de estudio mostraron valores de actividad de agua más bajos y una mejora en la textura y el color de los filetes comparado con los peces alimentados con la dieta control. La inclusión de remolacha y betaína no tuvo ningún efecto sobre los parámetros oxidativos del filete, mientras que la inclusión de cebada inhibió su oxidación lipídica. Concentraciones de cebada por encima del 8% mejoraron la actividad antioxidante de los filetes y aumentaron el contenido en alpha-tocoferol, sin embargo, los peces alimentados con remolacha y betaína mostraron un mayor contenido en flavonoides y fenólico pero no se observó ningún efecto sobre las propiedades antioxidantes del filete. Por último, cuando se llevó a cabo el análisis sensorial se observó que los peces alimentados con dietas con más de un 8% de cebada mostraron unas agallas más rojas y mejor textura, además de un color rojo del filete más intenso, parámetros que se correlacionan con una mejora de la frescura del pescado. Sin embargo, la inclusión de cebada o remolacha en dietas de trucha arco iris no tuvo ningún efecto sobre la aceptabilidad de los filetes. / The current Doctoral Thesis used barley and red beet as alternative carbohydrate sources in rainbow trout diets. The aim was to evaluate their effect on productive, histological and morphometric parameters, their effect on biochemical indexes after an acute stress and their effect on the quality of rainbow trout (Oncorhynchus mykiss). The different markers studied were the same in the barley and red beet experiments. In the barley experiment, increasing concentrations of barley (0-32%) were used in the diet, starting with an average weight of 127.72 ± 5.65 g and ending when they reached commercial weight after 84 days. In the red beet experiment, two red beet levels (14% and 28%) and two betaine levels (0.9% and 1.63%) were used, starting with an average weight of 69 ± 2.2 g and finishing when they reached commercial weight at the end of 105 days. The inclusion of barley in rainbow trout diets did not show significant differences in growth and biometric parameters. While the inclusion of 14% of red beet and 0.9% betaine did not affect growth, nutritive or biometric parameters and nutrient retentions compared to control, higher red beet and betaine concentrations showed a negative effect. When the histological and morphometric parameters were studied in liver and intestine, the results showed that fish fed at high barley concentrations showed smaller hepatocytes than control, while hepatocytes were larger in fish fed with red beet and betaine than control, however, in both cases, the liver had a lower level of vacuolization. Barley inclusion at 8% produced a negative effect at intestine morphological level; however, no effects were observed with the inclusion of red beet. Fish were submitted to stress, a lack of oxygen and increased of fish density, during 10 minutes. The results showed that at basal levels, the inclusion of barley and red beet did not cause any change in cortisol, glucose and MDA levels, although basal lactate values were significantly lower in the fish fed with red beet and betaine. Thirty minutes after stress the concentration of cortisol, glucose and lactate increased significantly in the fish of all groups and the concentration of MDA decreased. The inclusion of barley showed lower cortisol, glucose and lactate values than control. When fish reached commercial weight, the quality of the fillets and antioxidant properties were analysed. In both trials, fish fed with the experimental ingredients showed lower water activity values and textural and colour properties were enhanced. Red beet and betaine inclusion did not show any effect on the oxidative parameters of the fillets, while the inclusion of barley showed an inhibitory effect on fillets lipid oxidation. Concentrations of barley above 8% improved the antioxidant activity of the fillets and increased the content of alpha-tocopherol, however, fish fed with red beet and betaine showed a higher content of flavonoids and phenolics but no effect on the antioxidant properties of the fillet. Finally, when the sensory analysis was carried out, it was observed that fish fed diets with more than 8% barley showed redder gills and better texture than control, in addition to a more intense red colour of the fillet, these characteristics are correlated with an improvement of fish freshness. However, the inclusion of barley or red beet in rainbow trout diets had no effect on the acceptability of fillets. / En la present tesi doctoral es van utilitzar ordi i remolatxa com a fonts alternatives de carbohidrats en dietes per a truita amb l'objectiu d'avaluar el seu efecte sobre paràmetres productius, histologia hepàtica i intestinal, estrès i qualitat de la carn. Els marcadors estudiants fòren els mateixos en els experiments d`ordi I remolatxa. En les proves de l'ordi es van utilitzar concentracions creixents d'aquest ingredient (0-32%), es va iniciar amb un pes mitjà de 127.72 ± 5.65 g i es va finalitzar quan van arribar al pes comercial al cap de 84 dies. En les proves de remolatxa es van utilitzar dos nivells de remolatxa (14% i 28%) i dos de betaïna (0.9% i 1.63%), es va iniciar amb un pes mitjà de 69 ± 2.2 g i es va finalitzar quan els peixos van aconseguir el pes comercial al cap de 105 dies. La inclusió d'ordi en dietes de truita no va mostrar diferències significatives en els paràmetres de creixement i biomètrics. La inclusió de 14% remolatxa i 0,9% betaïna no va afectar el creixement, paràmetres nutritius, biomètrics i retencions de nutrients en comparació amb el control, concentracions majors de remolatxa i betaïna van tenir un efecte negatiu. Quan es van estudiar els paràmetres histològics i morfomètrics en fetge i intestí, els resultats van mostrar que els peixos alimentats amb concentracions creixents d'ordi tenien hepatòcits més petits, mentres que els peixos alimentats amb remolatxa i betaïna presentaven hepatòcits més grans, i en ambdós casos el fetge va mostrar un menor nivell de vacuolització. La concentracion d'ordi d'un 8% va produir un efecte negatiu a nivell morfològic de l'intestí, mentres que la inclusió de remolatxa no va produir cap efecte a nivell d'intestí. Els peixos van ser sotmesos a estrès per absència d'oxigen i augment de densitat durant 10 minuts. Quan l'estrès va ser analitzat a nivell bioquímic els resultats van mostrar que a nivells basals, la inclusió d'ordi i remolatxa no va causar cap canvi en els nivells de cortisol, glucosa i MDA, encara que els valors basals de lactat van ser significativament més baixos en els peixos alimentats amb remolatxa i betaïna. 30 minuts després de l'estrès la concentració de cortisol, glucosa i lactat va augmentar significativament en els peixos de tots els grups i la concentració de MDA va disminuir. La inclusió d`ordi va mostrar valors més baixos de cortisol, glucosa i lactat que la dieta control. Quan els peixos van aconseguir el pes comercial es va analitzar la qualitat dels filets i propietats antioxidants. En ambdues proves els peixos alimentats amb els ingredients objecte d'estudi van mostrar valors d'activitat d'aigua més baixos i una millora en la textura i el color dels filets comparat amb els peixos alimentats amb la dieta control. La inclusió de remolatxa i betaïna no va tenir cap efecte sobre els paràmetres oxidatius del filet, mentre que la inclusió d'ordi va inhibir l'oxidació lipídica dels filets. Concentracions d'ordi per sobre del 8% van millorar l'activitat antioxidant dels filets i van augmentar el contingut en alpha-tocoferol, però, els peixos alimentats amb remolatxa i betaïna van mostrar un major contingut en flavonoides i fenòlic però no es va observar cap efecte sobre les propietats antioxidants del filet. Finalment, quan es va dur a terme l'anàlisi sensorial es va observar que els peixos alimentats amb dietes amb més d'un 8% d'ordi van mostrar unes ganyes més vermelles i millor textura, a més d'un color vermell del filet més intens, paràmetres que es correlacionen amb una millora de la frescor del peix. No obstant això, la inclusió d'ordi o remolatxa en dietes de truita no va tenir cap efecte sobre l'acceptabilitat dels filets. / Pinedo Gil, J. (2018). Study of novel carbohydrate sources on rainbow trout (Oncorhynchus mykiss) diets [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/107364 / TESIS