Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-10-23T19:44:00Z No. of bitstreams: 2 Dissertação - Evandro Leite Bitencourt - 2015.pdf: 4223075 bytes, checksum: fe60d827b473e79c5c9c8ab601f67a69 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-10-23T19:46:10Z (GMT) No. of bitstreams: 2 Dissertação - Evandro Leite Bitencourt - 2015.pdf: 4223075 bytes, checksum: fe60d827b473e79c5c9c8ab601f67a69 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-10-23T19:46:10Z (GMT). No. of bitstreams: 2 Dissertação - Evandro Leite Bitencourt - 2015.pdf: 4223075 bytes, checksum: fe60d827b473e79c5c9c8ab601f67a69 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-03-31 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Mushrooms have been valued as a functional food and dietary supplement for humanity, due to the presence of substances with high nutritional value and/or therapeutic. Among the mushrooms recognized for its therapeutic properties we have is the leafy Grifola, popularly known as “Maitake”, which has important pharmacological actions such as anti-tumor, immunomodulatory, antiinflammatory, antioxidant, and others, which have been mainly related to its carbohydrates. Nevertheless, this study aimed to review the chemistry of the polysaccharides from mushroom of the two lines ("white" and "gray", represented as Gfb and Gfc, respectively) of G. frondosa leafy that have been cultivated in Brazil. From these type it was obtained branched -glucans containing links of type (13) (16) and heteropolysaccharides formed mainly by galactose (heterogalactan) or mannose (heteromannan). The heterogalactan composed mainly of fucose (15.7%), mannose (20.5%) and galactose (63.8%) was denominated fucomannogalactan (FMG-Gfb). It contains a main chain consisting of units of α-D-Galp and 3-Me-O-α-D-Galp connected (16), which are present partially substituted by O-2 disaccharide 3-O- α-D-Manp-α-L-Fucp to a lesser extent with terminal non-reducing α-L-Fucp and α-D-Manp. The heteromannan composed of fucose (6.9%), xylose (33.1%) and mannose (60.0%), a fucoxylomannan (FXM-Gfc) was found to be composed of a backbone of units α-D-Manp (13) connected, and a part of them substituted by O-3 side chains made of fucose and xylose. The results of this study suggest that there are no significant differences between the carbohydrate of both strains, showing that white strain it provide the therapeutic effects attributed to carbohydrates of another strain. / Os cogumelos têm sido valorizados como alimento funcional e suplemento alimentar para a humanidade, devido à presença de substâncias com alto valor nutricional e/ou medicinal. Dentre os cogumelos reconhecidos pelas suas propriedades terapêuticas encontra-se o Grifola frondosa, popularmente conhecido como Maitake, o qual possui importantes ações farmacológicas como antitumoral, imunomoduladora, anti-inflamatória, antioxidante, entre outras, as quais têm sido relacionadas, principalmente, aos seus carboidratos. Diante do exposto, o presente trabalho teve como objetivo a avaliação química dos polissacarídeos dos basidiomas das duas linhagens (branca e cinza, representadas como Gfb e Gfc, respectivamente) de G. frondosa que vem sendo cultivadas no Brasil. A partir destes foram obtidas as -glucanas ramificadas contendo ligações do tipo (13) (16) e os heteropolissacarídeos formados principalmente por galactose (heterogalactana) ou manose (heteromanana). A heterogalactana constituída, principalmente, por fucose (15,7%), manose (20,5%) e galactose (63,8%), foi denominada de fucomanogalactana (FMG-Gfb). Esta contém uma cadeia principal formada por unidades de α-D-Galp e de 3-O-Me-α-D-Galp ligadas (16), as quais se apresentam parcialmente substituídas em O-2 pelo dissacarídeo 3-O-α-DManp- α-L-Fucp e em menor proporção com terminais não redutores de α-LFucp e α-D-Manp. A heteromanana, composta por fucose (6,9%), xilose (33,1%) e manose (60,0%), ou seja, uma fucoxilomanana (FXM-Gfc) mostrou ser constituída por uma cadeia principal composta por unidades de α-D-Manp (13) ligadas, sendo uma parte destas substituídas em O-3 por cadeias laterais formadas por fucose e xilose. Os resultados obtidos neste estudo sugerem que não há diferenças significativas entre os carboidratos de ambas as linhagens, evidenciando que a linhagem branca deve apresentar os efeitos terapêuticos atribuídos aos carboidratos da outra linhagem.

Les objectifs de ce projet de maitrise sont d'analyser la composition d'un coproduit de l'holothurie Cucumaria frondosa et de déterminer ses effets sur des paramètres cardiométaboliques reconnus quand administré par voie orale. Afin d'atteindre ce dernier objectif, quatre groupes de 10 rats Wistar mâles chacun ont été nourris pendant 28 jours soit avec une diète contrôle purifiée, soit avec une diète enrichie avec le coproduit à 0,5%, 1,5% ou 2,5% de protéines de concombre de mer. Les fèces ont été collectées aux jours 21, 22 et 23. Les rats ont été sacrifiés après un jeûne de 10 heures au jour 29. Le sang a été prélevé par ponction aortique afin d'obtenir du plasma et du sérum. Le poids des tissus adipeux (AT) épididymaux, périrénaux et rétropéritonéaux, de l'extensor digitorum longus (EDL), le soléaire, des glandes surrénales, du foie, ainsi que la longueur des intestins ont été mesurés. Le foie et les tissus adipeux ont ensuite été aliquotés, surgelés et conservés à -80°C pour analyses futures. La composition corporelle a été déterminée à l'aide d'une bombe calorimétrique pour déterminer l'énergie totale et avec une unité d'analyse Foss Kjeltec 8400 pour déterminer la masse maigre. Une extraction Folch modifiée et des kits de test enzymatique colorimétrique ont été utilisés pour mesurer le cholestérol et les triglycérides (TG) hépatiques. Des kits de test enzymatique colorimétrique et immunométrique ont été utilisés pour déterminer les taux de cholestérol à lipoprotéines de haute densité (HDL-C), de cholestérol à lipoprotéines de très basse densité et de basse densité (VLDL + LDL-C), de TG et d'insuline plasmatiques. Le contenu en lipides fécaux a également été mesuré. La taille des adipocytes épididymaux a été mesurée en utilisant des lames d'histologie colorées à l'hématoxyline et à l'éosine (H&E) de l'AT épididymal. Le coproduit de l'holothurie contenait des concentrations élevées de plusieurs nutriments, notamment des protéines, des acides aminés essentiels, des lipides, en particulier des phospholipides et des acides gras polyinsaturés (PUFA) oméga-3, du fer, du potassium, du sodium, du phosphore, du calcium, du sélénium et du zinc. À l'inverse, le coproduit était pauvre en gras saturés, en cholestérol, en PUFA oméga-6 et en glucides. De plus, le coproduit contenait de grandes quantités de composés bioactifs, en particulier des saponines. Par ailleurs, la teneur en arsenic du coproduit, surtout en arsenic inorganique dont une ingestion prolongée peut entraîner des effets indésirables sur la santé, était élevée dans le coproduit. Quant au protocole animal, le coproduit n'a pas influencé la prise alimentaire ni le poids corporel final des animaux. Le groupe à 1,5% avait des adipocytes épididymaux plus petits (P=0.03) que le groupe témoin. Cette diminution suggère que l'administration orale du coproduit à une dose de 1,5% peut diminuer l'adiposité viscérale, indiquant un potentiel bénéfique sur la santé cardiométabolique (CMH). Ces effets observés peuvent être dus à la teneur en saponine du coproduit, mais les effets des autres composants ne peuvent être exclus. À partir de ces résultats, les coproduits d'holothuries et les saponines ont un potentiel dans la prévention des maladies cardiométaboliques, mais des recherches supplémentaires sont nécessaires pour déterminer l'efficacité et l'innocuité du coproduit. / The objectives of this master's project were to analyse the chemical and nutritional composition of a sea cucumber coproduct prepared from the species Cucumaria frondosa and to determine the effects of said coproduct on recognised cardiometabolic parameters when administered orally. In order to achieve the latter objective, four groups of 10 male Wistar rats each were fed for 28 days either a purified control diet or a diet enriched with the coproduct at 0.5%, 1.5% and 2.5% of sea cucumber protein to total protein content. Feces were collected on day 21, 22, and 23. The rats were sacrificed on day 29 after a 10 hour fast. Blood was collected by aortic puncture in order to obtain plasma and serum samples. The weight of the epididymal, perirenal, and retroperitoneal adipose tissues (AT), the extensor digitorum longus (EDL), the soleus, the suprarenal glands, the liver, as well as the length of the intestines were recorded. The liver and adipose tissues were then aliquoted, flash frozen, and stored at -80°C for future analysis. Body composition was calculated using a calorimetric bomb to determine total corporeal energy and a Foss Kjeltec 8400 analyser unit to determine crude protein. A modified Folch extraction and enzymatic colorimetric assay kits were used to measure hepatic cholesterol and triglycerides (TG). Enzymatic colorimetric and immunometric assay kits were also used to determine levels of serum high-density lipoprotein cholesterol (HDL-C), serum very low density and low-density lipoprotein cholesterol (VLDL+LDL-C), plasma TG, and plasma insulin, respectively. The lipid content of the collected feces was also determined. Using hematoxylin and eosin (H&E)-stained histology slides of the epididymal AT, the size of the epididymal adipocytes was measured. The analyses of the coproduct revealed that it contained high concentrations of several nutrients including protein, essential amino acids, lipids, namely phospholipids and omega-3 poly unsaturated fatty acids (PUFAs), iron, potassium, sodium, phosphorus, calcium, selenium, and zinc. Conversely, the coproduct was low in saturated fat, cholesterol, omega-6 PUFAs, and carbohydrates. Furthermore, the coproduct contained high amounts of bioactive compounds, particularly saponins. However, the arsenic content of the coproduct, particularly inorganic arsenic, was high. Regarding the animal protocol, the addition of the coproduct did not influence food intake or final body weight. The 1.5% group had smaller epididymal adipocytes (P=0.03) than the control group. This decrease in adipocyte size suggests that oral administration of the coproduct at a dose of ≈1.5% can decrease visceral adiposity, indicating the potential to benefit cardiometabolic health (CMH). These observed effects may be attributable to the saponin content of the coproduct, but the effects of other components cannot be ruled out. From these results, sea cucumber coproducts and saponins have potential in the prevention of cardiometabolic diseases, but further research is necessary to determine the efficacy and innocuity of the coproduct.

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.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2014-11-19T13:28:06Z No. of bitstreams: 2 Dissertação - Gracy Kelly Faria Oliveira - 2013.pdf: 3056383 bytes, checksum: 704c745d35b0898a2ea697e0aebcc431 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-11-20T14:24:58Z (GMT) No. of bitstreams: 2 Dissertação - Gracy Kelly Faria Oliveira - 2013.pdf: 3056383 bytes, checksum: 704c745d35b0898a2ea697e0aebcc431 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-11-20T14:24:58Z (GMT). No. of bitstreams: 2 Dissertação - Gracy Kelly Faria Oliveira - 2013.pdf: 3056383 bytes, checksum: 704c745d35b0898a2ea697e0aebcc431 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-07-18 / Partially methylated heterogalactans were isolated from the fruiting bodies of the medicinal mushroom Pleurotus ostreatus (mannogalactan) and Grifola frondosa (mannofucogalactan), via successive cold aqueous extraction, followed by fractionation through freeze-thawing, precipitation with Fehling solution of soluble material, and dialysis using membranes with different exclusion limits (MWCO 50, 100, and 500 kDa). Their chemical structures were established based on monosaccharide composition, methylation analysis and NMR studies. The mannogalactan (Mw = 50.7 x 103 g.mol-1) had a main chain of (1→6)-linked α-D-galactopyranosyl and 3-O-methyl-α-D-galactopyranosyl units, both of which are partially substituted at O-2 by -D-mannopyranosyl nonreducing ends.The other was a mannofucogalactan (Mw = 15.9 x 103 g.mol-1), which had a similar main-chain, being a part of the galactopyranosyl units substituted at O-2 mainly by 3-O-α-D-mannopyranosyl-α-L-fucopyranosyl groups and in a minor proportion with α-L-Fucp single-unit side chains. Recently, the heterogalactans are also recognized for their relevant biological activities, such as antitumor, immunomodulatory, anti-inflammatory and antinociceptive effects. Thus, the heterogalactans obtained from G.frondosa and P. ostreatus could present itself as a good candidate to be evaluated for its biological potential. / Heterogalactanas parcialmente metiladas foram isoladas dos basidiomas dos cogumelos medicinais Pleurotus ostreatus (manogalactana) e Grifola frondosa (manofucogalactana), através de sucessivas extrações aquosas a frio, seguidas de fracionamento por congelamento e degelo, precipitação com solução de Fehling e diálises em membranas com diferentes limites de exclusão (MWCO 50, 100 e 500 kDa). As estruturas químicas foram determinadas por meio das análises de composição monossacarídica, metilação e RMN (mono e bidimensionais). A manogalactana (Mw = 50,7 x 103 g.mol-1) apresentou uma cadeia principal constituída por unidades de -D-Galp e 3-O-Me--D-Galp (16) ligadas, sendo ambas parcialmente substituídas em O-2 por terminais não redutores de -D-Manp. A outra foi uma manofucogalactana (Mw = 15,9 x 103 g.mol-1), a qual apresentou uma cadeia principal similar, sendo uma parte das unidades de -D-galactopiranosil substituídas em O-2 pelo dissacarídeo 3-O-α-D-manopiranosil-α-L-fucopiranosil e em menor proporção com terminais não redutores de α-L-Fucp. Recentemente, as heterogalactanas têm sido reconhecidas pelas suas ações biológicas relevantes, como a atividade antitumoral, imunomodulatória, antinociceptiva e/ou anti-inflamatória. Sendo assim, as heterogalactanas isoladas de G. frondosa e P. ostreatus podem apresentar-se como bons candidatos a serem avaliados quanto ao seu potencial biológico.

Universiẗat, Diss., 2001--Jena.

<p>Sea cucumbers have been found to contain bioactive compounds such as saponin, fucoidan, frondoside, and glycosides that have pharmacological properties like antitumor, antibacterial, anti-inflammation, and antihyperglycemic activity. Although several species of sea cucumbers have been studied and reared for the food and medicinal industries, not much research has been conducted on the species in the waters of the Eastern United States. In this research, physiological and immunological parameters of coelomic fluid from <em>Cucumaria</em> <em>frondosa</em>, <em>Isostychopus</em> <em>badionotus</em>, and <em>Pentacta</em> <em>pygmaea</em> were compared to find the most promising candidate with these properties and pharmacological benefits. We found that <em>C. frondosa</em> was the species with the best immunological and physiological parameters among the three studied. <em>C. frondosa</em> illustrated that its coelomic fluid contains the highest concentrations of cells and lysozymes that had the highest activity. Using <em>C. frondosa</em>’s tissue extracts and coelomic fluid, the ability of the extracts and coelomic fluid to inhibit murine melanoma cells (B16-F10) and modulate T-lymphocytes <em>in vitro</em> were investigated. Although no significant differences were seen statistically, the experiments illustrated that T-lymphocytes were highly activated at higher concentrations (0.001g/uL-0.0002g/uL) for tissue extracts and at lower concentrations (0.000008g/uL) for coelomic fluid. On the other hand, melanoma cells were inhibited highest at lower concentrations (0.000008g/uL-0.0000016/uL). In addition to these studies, the antibacterial activity of <em>C. frondosa</em> extract was tested on ten pathogenic bacterial species. Antibacterial activity of the <em>C. frondosa</em> extract was not seen in this experiment. However, hemolytic activity by compounds present in <em>C. frondosa</em> extracts was seen in blood agars culturing <em>Streptococcus pneumoniae</em> and <em>Enterococcus faecalis</em> in our experiment. Lastly, an <em>in vivo </em>study was conducted to see if <em>C. frondosa</em> extract can modulate stress in Nile tilapia. In our experiment, we observed that <em>C. frondosa</em> extract was able to enhance the activity of one of the parameters, phagocytic capacity significantly. However, we are not able to conclude that <em>C. frondosa</em> extract was able to mitigate chronic stress from the results obtained. Overall, observing the results from the projects, we cannot conclude that <em>C. frondosa</em> extracts illustrated pharmacological properties. Extensive studies are recommended and required to use <em>C. frondosa</em> extract for medicinal purposes. </p>