I. Psilocybe cubensis was cultured successfully in two media. Medium A was devised by Catalfomo and Tyler and Medium B was a modification of a medium which has been used for ergot alkaloid production by Claviceps purpurea. Only when the fungus was kept on Sabouraud agar plates.did it subsequently produce psilocybin when transferred to liquid media. A quantitative time-course study of psilocybin production in the two media was carried out. Maximal production appeared on the fifth day. The activities of an acid phosphatase, acting on psilocybin, were measured from mycelia grown in the two media. Enzyme activity from the A culture was very high and a blue color caused by oxidation of psilocin formed in five minutes. The effect of adding L-tryptophan on alkaloid production as well as the fate of tryptophan-C¹⁴ was also investigated. Tryptophan stimulated significantly psilocybin production in the very beginning in the B medium. The degradation of tryptophan was different in the two media. It was converted to kynurenine and anthranilic acid in A medium and to tryptamine in tryptophan added B medium (B' medium). Radioactive D,L-tryptophan side chain labeled, gave labeled psilocin and psilocybin. Potassium deficiency decreased psilocybin production while a potassium supplement had no effect. The fungus did not produce polyacetylenic compounds in the medium but ergosterol was detected as a major acetate derived metabolite when the fungus was kept on MYP agar plates and transferred subsequently to liquid media. Psilocin has very slight antibiotic activity against Candida albicans whereas psilocybin has none. II. Eight species of Lentinus and Lentinellus were investigated for the occurrence of a phenol-O-methyltransferase. Only Lentinus lepideus and Lentinus pbnderbsus showed enzyme activity in both light and dark conditions. The specificity of the enzyme for a number of substrates was also examined. Of six compounds tested, methyl p-coumarate, methyl caffeate and methyl ferulate.served-as substrates. The products of enzymic activity were identified-by radioautography. / Science, Faculty of / Botany, Department of / Graduate

Cheng Chiu-Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 112-123). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.III / 摘要 --- p.V / LIST OF FIGURES --- p.XII / LIST OF TABLES --- p.XV / LIST OF ABBREVIATIONS --- p.XVII / Chapter CHAPTER 1 --- LITERATURE REVIEW --- p.1 / Chapter 1.1 --- General Introduction --- p.1 / Chapter 1.2 --- Cancer Treatment Modalities --- p.3 / Chapter 1.3 --- Cancer Immunotherapy --- p.4 / Chapter 1.3.1 --- Immunomodulation --- p.4 / Chapter 1.3.2 --- Conventional immunotherapy --- p.5 / Chapter 1.3.2.1 --- Active immunotherapy --- p.5 / Chapter 1.3.2.2 --- Passive immunotherapy --- p.5 / Chapter 1.3.3 --- Novel immunotherapy --- p.8 / Chapter 1.3.3.1 --- Natural Products from Chinese Medicinal Herbs as BRMs --- p.9 / Chapter 1.4 --- Antitumor polysaccharides from Mushrooms --- p.10 / Chapter 1.4.1 --- Composition of mushroom antitumor polysaccharides --- p.10 / Chapter 1.4.1.1 --- β-glucan --- p.10 / Chapter 1.4.1.2 --- Heteroglucan --- p.11 / Chapter 1.4.1.3 --- Glycan and polysaccharide-protein complexes --- p.11 / Chapter 1.4.2 --- Antitumor polysaccharides from Polyporaceae mushrooms --- p.14 / Chapter 1.4.2.1 --- Lentinan from Lentinus edodes --- p.16 / Chapter 1.4.2.2 --- Maitake from Grifola frondosa --- p.17 / Chapter 1.4.2.3 --- Polysaccharides from Ganoderma lucidum --- p.18 / Chapter 1.4.3 --- Extraction and purification of Polysaccharides from Mushrooms --- p.19 / Chapter 1.4.4 --- Chemical modification --- p.20 / Chapter 1.5 --- Polyporus umbellatus (Zhuling) --- p.20 / Chapter 1.5.1.1 --- Nutritional Contents --- p.21 / Chapter 1.5.1.2 --- Medicinal properties --- p.21 / Chapter 1.6 --- The Objective of the Present Project --- p.23 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.25 / Chapter 2.1 --- Extraction --- p.25 / Chapter 2.1.1 --- Hot-water extraction of crude Polyporus umbellatus polysaccharides --- p.25 / Chapter 2.1.2 --- Cold-alkaline extraction of crude Polyporus umbellatus polysaccharides --- p.26 / Chapter 2.2 --- Purification of crude Polyporus umbellatus polysaccharides --- p.29 / Chapter 2.2.1 --- Preparation of DEAE-cellulose ion exchange column --- p.29 / Chapter 2.2.2 --- Fractionation of hot-water soluble crude polysaccharide (PU) --- p.29 / Chapter 2.3 --- Characterization of Polyporus umbellatus Polysaccharides --- p.30 / Chapter 2.3.1 --- Carbohydrate content determination --- p.30 / Chapter 2.3.2 --- Gas chromatography (GC) --- p.31 / Chapter 2.3.3 --- Protein content determination --- p.32 / Chapter 2.3.4 --- Uronic acid content determination --- p.33 / Chapter 2.3.5 --- High Performance Liquid Chromatography (HPLC) --- p.34 / Chapter 2.4 --- In vivo Antineoplastic Assay --- p.35 / Chapter 2.4.1 --- Animals --- p.35 / Chapter 2.4.1.1 --- BALB/c mice --- p.35 / Chapter 2.4.1.2 --- Athymic BALB/c mice --- p.36 / Chapter 2.4.2 --- Maintenance of cell lines --- p.36 / Chapter 2.4.2.1 --- Murine sarcoma 180 --- p.36 / Chapter 2.4.2.2 --- Human breast cancer cell line (MCF-7) --- p.37 / Chapter 2.4.3 --- S-180 tumor inoculation model using BALB/c mice --- p.37 / Chapter 2.4.4 --- MCF-7 tumor inoculation model using athymic nude mice --- p.38 / Chapter 2.4.5 --- Assay of antineoplastic activity with S-180 --- p.38 / Chapter 2.4.6 --- Assay of antineoplastic activity with MCF-7 --- p.39 / Chapter 2.4.7 --- Body weight change --- p.40 / Chapter 2.5 --- In vitro anti-proliferation assay --- p.41 / Chapter 2.5.1 --- Cell lines --- p.41 / Chapter 2.5.1.1 --- Maintenance of cell lines --- p.41 / Chapter 2.5.2 --- Assay of anti-proliferation with cancer cell lines --- p.42 / Chapter 2.5.2.1 --- Cytotoxicity assay on suspensions of cancer cells --- p.42 / Chapter 2.5.2.2 --- Cytotoxicity assay on adhesive cancer cells --- p.42 / Chapter 2.5.2.3 --- Cytotoxicity assay on normal cells --- p.43 / Chapter 2.5.3 --- Trypan blue exclusion method --- p.43 / Chapter 2.5.4 --- MTT assay --- p.44 / Chapter 2.6 --- Cytokine determination --- p.45 / Chapter 2.6.1 --- Treatment of mice --- p.45 / Chapter 2.6.2 --- Enzyme-linked immunosorbent assay (ELISA) for TNF-a production --- p.46 / Chapter 2.6.3 --- Analysis of mouse cytokine array --- p.47 / Chapter 2.6.3.1 --- Process of blocking and incubation --- p.47 / Chapter 2.6.3.2 --- Process of cytokine detection --- p.48 / Chapter 2.7 --- Statistical Analysis --- p.49 / Chapter CHAPTER 3 --- RESULTS --- p.50 / Chapter 3.1 --- Extraction of Polyporus umbellatus polysaccharides (PU) --- p.50 / Chapter 3.1.1 --- Percentage of Yield in extraction of crude PU extracts --- p.50 / Chapter 3.1.2 --- Percentage of yield in fractionation of PU fractions --- p.51 / Chapter 3.2 --- Chemical characterization of PUW fractions --- p.55 / Chapter 3.2.1 --- Carbohydrate and protein contents of puw fractions --- p.55 / Chapter 3.2.2 --- Relative content of monosaccharides and uronic acid in PUW fractions --- p.55 / Chapter 3.2.3 --- Infrared spectra of PUW fractions --- p.59 / Chapter 3.2.4 --- Molecular weight estimation of PUW fractions --- p.59 / Chapter 3.3 --- In vitro anti-proliferative assay --- p.64 / Chapter 3.3.1 --- Anti-proliferative effects of PU fractions on suspension cancer cell lines --- p.64 / Chapter 3.3.2 --- Anti-proliferative effects of PU fractions on adhesive cancer cell lines --- p.64 / Chapter 3.3.3 --- Anti-proliferative effects of PU fractions on normal cell lines --- p.65 / Chapter 3.4 --- In vivo antineoplastic assay --- p.73 / Chapter 3.4.1 --- S-180 tumor inoculation model using BALB/c mice --- p.73 / Chapter 3.4.1.1 --- In vivo antineoplastic effect of crude extracts - PUW and PUAL --- p.73 / Chapter 3.4.1.2 --- In vivo antineoplastic effect of PUW --- p.73 / Chapter 3.4.1.3 --- In vivo antineoplastic effect of PU60 and PU80 --- p.78 / Chapter 3.4.1.4 --- In vivo antineoplastic effect of PU60a and b --- p.81 / Chapter 3.4.2 --- MCF-7 tumor inoculation model using athymic nude mice --- p.81 / Chapter 3.4.2.1 --- In vivo antineoplastic effect of PU60b --- p.81 / Chapter 3.4.3 --- Identification of cytokines in the serum from healthy BALB/c mice using mouse cytokine array system --- p.89 / Chapter 3.4.4 --- Effect of PU60b on TNF-α generation in healthy BALB/c mice studied with ELISA. --- p.93 / Chapter 3.4.5 --- Effect of PU60b on TNF-α generation in S-180 tumor bearing mice studied with ELISA --- p.93 / Chapter CHAPTER 4 --- DISCUSSION --- p.96 / Chapter 4.1 --- Extraction and Isolation of polysaccharide fractions --- p.96 / Chapter 4.2 --- Chemical characterization of PUw fractions --- p.97 / Chapter 4.3 --- In vitro anti-proliferative effect of PU fractions --- p.99 / Chapter 4.4 --- In vivo antineoplastic assay of PU fractions --- p.101 / Chapter 4.4.1 --- S-180 solid tumor model using BALB/c mice --- p.101 / Chapter 4.4.2 --- MCF-7 tumor in vivo model using athymic nude mice --- p.104 / Chapter 4.5 --- Estimation of Immunomodulatory properties of PU60b --- p.106 / Chapter 4.5.1 --- Identification of cytokines in serum from healthy BALB/c mice --- p.106 / Chapter CHAPTER 5 --- CONCLUSIONS --- p.110 / REFERENCES --- p.112

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas. Programa de Pós-Graduação em Biologia Vegetal. / Made available in DSpace on 2012-10-24T10:26:14Z (GMT). No. of bitstreams: 1 267106.pdf: 7769043 bytes, checksum: e76b30fe652ca3fb23d1999aaacc3ffa (MD5) / O levantamento dos basidiomicetes (Basidiomycota, Fungi) lignolíticos no município de Mondaí, Santa Catarina, Brasil, resultou na identificação de 45 espécies distribuídas nas famílias Dacrymycetaceae J. Schröt. (1), Gloeophylaceae Jülich (1), Hymenochaetaceae Imazeki & Toki (14), Schizophylaceae Jülich (1), Schizoporaceae Quél. (2), Ganodermataceae Donk. (3), Meripilaceae Jülich (5), Meruliaceae P. Karst (3), Polyporaceae Fr.ex Corda (14) e Stecherinaceae Parmasto (1), pertencentes a 4 ordens (Dacrymycetales, Gloeophyllalles, Hymenochaetales e Polyporales) e a 2 classes (Dacrymycetes e Agaricomycetes). Uma espécie, Phellinus garuhapensis Wright & Blumenf foi citada pela primeira vez para o Brasil. Por outro lado, Dacryopinax elegans (Berk. & Curtis) Martin, Hymenochaete rubiginosa (Dick.:Fr) Lév., Inonotus rickii (Pat.) Reid, Phellinus rhytiphloeus (Mont.) Ryvarden, Phylloporia pectinata (Klotzsch) Ryvarden, Echinoporia aculeifera (Berk. & M.A. Curtis) Ryvarden, Oxyporus obducens (Pers.) Donk, Amauroderma sprucei (Pat.) Torrend e Pseudofavolus miquelii (Mont.) Pat. são espécies citadas pela primeira vez para o Estado de Santa Catarina. Todas as espécies foram novos registros para Mondaí. Das 45 espécies, todas são causadoras de podridão branca, com exceção de Stiptophyllum erubescens (Berk.) Ryvarden.

Submitted by Fernanda Rodrigues de Lima (fernanda.rlima@ufpe.br) on 2018-08-21T22:37:59Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Nelson Correia de Lima Júnior.pdf: 4273136 bytes, checksum: 4271b4fee1ae4a971ff8f1a57f0b2f9e (MD5) / Approved for entry into archive by Alice Araujo (alice.caraujo@ufpe.br) on 2018-08-28T22:41:07Z (GMT) No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Nelson Correia de Lima Júnior.pdf: 4273136 bytes, checksum: 4271b4fee1ae4a971ff8f1a57f0b2f9e (MD5) / Made available in DSpace on 2018-08-28T22:41:07Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Nelson Correia de Lima Júnior.pdf: 4273136 bytes, checksum: 4271b4fee1ae4a971ff8f1a57f0b2f9e (MD5) Previous issue date: 2016-02-25 / CNPq / Hymenochaetaceae Donk (Basidiomycota, Fungi) caracteriza-se por apresentar basidiomas ressupinados a pileados, reação xantocróica positiva, presença de setas himeniais (na maioria) e de hifas generativas sem grampos de conexão, além de reunir fungos causadoras de podridão branca. A circunscrição dos gêneros e espécies em Hymenochaetaceae é imprecisa, principalmente no âmbito da taxonomia tradicional, o que torna dúbia a identificação de espécies quando são utilizados exclusivamente dados morfológicos. Nessa perspectiva, estudos baseados em sequências de DNA das regiões ITS e LSU (rDNA) estão sendo responsáveis por significativos progressos na taxonomia do grupo, porém estudos com táxons neotropicais são escassos. No presente estudo foram utilizados espécimes de Hymenochaetaceae coletados em diferentes biomas brasileiros para extração de DNA genômico, amplificação das regiões ITS e LSU por meio de PCR e posterior sequenciamento. Foi observado o delineamento morfológico e/ou molecular de duas espécies de Fomitiporia, duas de Fulvifomes, quatro de Fuscoporia, seis de Hymenochaete, duas de Phellinotus, uma de Phellinus e uma de Tropicoporus. Dentre essas, são propostas: uma nova espécie de Fuscoporia; uma sinonimização em Phellinus, uma em Tropicoporus, uma em Hymenochaete e uma possível sinonimização em Fomitiporia; além da confirmação de combinação Fulvifomes rhytiphloeus a partir de evidências moleculares. São registradas ainda uma nova ocorrência de Fomitiporia e de Hymenochaete para o Brasil. De modo geral, observou-se que as sequências da região ITS e/ou LSU foram úteis no delineamento das espécies em estudo, principalmente em espécies morfologicamente semelhantes e próximas do ponto de vista evolutivo. / Hymenochaetaceae Donk (Basidiomycota, Fungi) is characterized by sessile or pileate basidiomes, positive xanthochroic reaction, occurrence of hymenial setae (frequent) and absence of clamps. Besides, species of this group are also known for producing white rot. The circumscription of genera and species within Hymenochaetaceae is inaccurate, especially under the traditional taxonomy, which makes dubious species identification when used exclusively morphological data. From this perspective, studies based on DNA sequences of the regions ITS and LSU (rDNA) are accounting for significant progress in the taxonomy of the group, but studies with Neotropical taxa are scarce. In this study, Hymenochaetaceae specimens were collected in different Brazilian biomes for DNA extraction, amplification of the regions ITS and LSU by PCR and subsequent sequencing. We observed the morphological and/or molecular delimitation of two species of Fomitiporia, two of Fulvifomes, four of Fuscoporia, seven of Hymenochaete, two of Phellinotus, one of Phellinus and one of Tropicoporus. Among these it is proposed: a new species of Fuscoporia; one synonymization in Phellinus, one in Tropicoporus, one in Hymenochaete and a possible synonymization in Fomitiporia; and a confirmation of the combination in Fulvifomes rhytiphloeus upon molecular data. We also report a new record of Fomitiporia and another of Hymenochaete from Brazil. In general, it was observed that sequences of ITS and/or LSU region are useful for species delimitation, particularly for those which are morphologically similar and evolutionarily close related.

Submitted by Pedro Barros (pedro.silvabarros@ufpe.br) on 2018-09-10T21:37:38Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Odacy Camilo de Souza.pdf: 2425125 bytes, checksum: 7bb72fbb05058f53be027a8e54ba439c (MD5) / Approved for entry into archive by Alice Araujo (alice.caraujo@ufpe.br) on 2018-09-17T18:12:14Z (GMT) No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Odacy Camilo de Souza.pdf: 2425125 bytes, checksum: 7bb72fbb05058f53be027a8e54ba439c (MD5) / Made available in DSpace on 2018-09-17T18:12:14Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Odacy Camilo de Souza.pdf: 2425125 bytes, checksum: 7bb72fbb05058f53be027a8e54ba439c (MD5) Previous issue date: 2015-06-30 / CAPES / Trichosporon são leveduras pertencentes ao filo Basidiomycota, que se destacam na produção de lipases, dentre outras enzimas. Os objetivos deste estudo foram Selecionar, culturas de Trichosporon mantidas na Micoteca URM, quanto à produção de lipases, através de avaliação qualitativa e quantitativa, otimizar a produção da enzima, caracterizar, pré-purificar e aplicar avaliando a capacidade de degradar efluentes de abatedouros de bovinos. Foram realizadas a reativação e autenticação das culturas através das características morfofisiológicas. Para degradação do meio rico em lipídeos, foram retirados fragmentos das culturas mantidas em SYE contidos em tubo de ensaio e transferidos para o centro da placa de Petri contendo o mesmo meio. Após sete dias de crescimento, fragmentos da cultura foram transferidos para o centro da placa de Petri com meio de cultura. As culturas foram incubadas a 28ºC e 37ºC, observadas durante 24, 48, 72 e 96 horas, para evidenciar a presença de halo opaco ao redor da colônia. Para determinação da capacidade lipolítica em fermentação submersa O experimento foi realizado utilizando Erlenmeyers de 250 mL contendo 50mL dos seguintes meios; I composto por 1% (v/v) de óleo de oliva, meio II contendo 1% (v/v) de óleo de mamona, e meio III composto por 1% (v/v) de óleo de licuri, adicionados de 0,5% (p/v) extrato de levedura e pH ajustado para 6,5. Esterilizados e inoculados e incubados a 160 rpm a 37ºC por 96 horas. Foi utilizado o método espectrofotométrico para determinar a atividade da lipase. Otimização do meio de produção de lipases por estatística de aproximação, o meio foi preparado utilizando diferentes concentrações de nutrientes testados de acordo com os experimentos do planejamento estatístico. Os inóculos foram adicionados a cada 50 mL de meio em Erlenmeyer de 250 mL de acordo com a concepção da matriz. Os frascos foram incubados por até cinco dias em diferentes temperaturas, sob agitação em diferentes rotações. Identificação e seleção das variáveis significativas para a produção de lipases foi realizada com base no design PB. Foram testados 11 variáveis. Para a Metodologia de superfície de resposta foram usadas as variáveis selecionadas para maior produção de lipase. Para caracterização enzimática foi avaliado o efeito e a estabilidade do pH, temperatura, íons, solventes e detergentes sobre a atividade enzimática. Na purificação foi utilizado o sistema de duas fases aquosas, com planejamento estatístico 2³, assim como a aplicação do estrato bruto. Em meio sólido, todas as culturas apresentaram crescimento, sem presença de halo de degradação, em cultivo submerso, todas as culturas apresentaram atividade. Trichosporon sporotrichoides URM 6630, produziu 104,07 UmL. O melhor pH e temperatura para produção da enzima foram pH 10.0 e 40 ˚C. A purificação teve coeficiente de partição 76 e recuperação da atividade 164,71%, e o fator de purificação de 0,96. Para a aplicação a melhor de taxa de degradação foi obtida com a menor concentração do extrato bruto. Primeiro relato de produção de lipases por essa espécie, e da utilização de óleo de licuri como substrato. Este micro-organismo torna-se bastante promissor para a produção de lipases produz altos níveis destas enzimas, em condições que minimizando os custos da produção. / Trichosporon yeasts are belonging to the phylum Basidiomycota, they are highlighted in lipase activity among other enzymes. The objectives of this study were to select, Trichosporon cultures kept in the URM Culture Collection, as lipase production, optimize production of the enzyme, characterize, purify and apply evaluating the ability to degrade cattle slaughterhouses wastewater. They were held reactivation and authentication of cultures through the morphological and physiological characteristics. Degradation of the lipase fragments were removed from cultures maintained in SYE contained in a test tube and transferred to the center of the petri dish containing said medium. After seven days of growth, the culture pieces were transferred to the center of the petri dish with culture medium. Cultures were incubated at 28 ° C and 37 ° C, observed for 24, 48, 72 and 96 hours, demonstrating an opaque halo around the colony. To determine the lipolytic capacity in submerged fermentation The experiment was conducted using 250 mL Erlenmeyer flasks containing 50ml of the following means; I consists of 1% (v / v) olive oil, medium II containing 1% (v / v) castor oil, and medium III consists of 1% (v / v) licuri oil added 0 5% (w / v) yeast extract and pH adjusted to 6.5. Sterilized and inoculated and incubated at 160 rpm at 37 ° C for 96 hours. The spectrophotometric method was used to determine the lipase activity. Optimization of lipase production medium by statistical approach, the medium was prepared using different concentrations of nutrients tested according to the statistical design of experiments. The inocula were added to each 50 ml of medium in 250 ml Erlenmeyer according to the design of the array. The flasks were incubated for five days at different temperatures under stirring at different speeds. Identification and selection of the variables significant for lipase production was based on the design PB. 11 variables were tested. For response surface methodology were used the variables selected for increased production of lipase. For enzyme characterization, the effects and stability on pH, temperature, ions, solvents and detergents on the enzymatic activity. Purification was used in the aqueous two-phase system with statistical design 2³, as well as the application of the crude stratum. In a solid medium, all cultures grew without the presence of degradation halo, in submerged cultures, all cultures showed activity. Trichosporon sporotrichoides URM 6630, produced 104.07 UML. The best pH and temperature for enzyme production were pH 10.0 and 40 ° C. Purification was 76 and partition coefficient 164.71% activity recovery, and purification factor of 0.96. To implement the best degradation rate was obtained with the lowest concentration of the crude extract. First report of lipase production by this species, and use of licuri oil as substrate. This micro-organism becomes quite promising for lipase production produces high levels of these enzymes under conditions which minimize production.

Submitted by Daniella Sodre (daniella.sodre@ufpe.br) on 2015-03-17T11:47:51Z No. of bitstreams: 2 Dissertação Lidia Silva.pdf: 4103104 bytes, checksum: eb5e0fe6bb40a7f4698d819d1e3c3755 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-17T11:47:51Z (GMT). No. of bitstreams: 2 Dissertação Lidia Silva.pdf: 4103104 bytes, checksum: eb5e0fe6bb40a7f4698d819d1e3c3755 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2013-02-22 / CNPq / Os Agaricomycetes de solo são representados pelos fungos clavarioides, estereoides, agaricoides e algumas espécies poroides. Esses fungos apresentam um papel fundamental na manutenção desses ecossistemas através da ciclagem de nutrientes e suas diferentes relações ecológicas (saprofitismo, parasitismo e micorrização). Com o objetivo de avaliar a diversidade e as interações ecológicas de Agaricomycetes terrícolas (clavarioides, estereoides e poroides), esses fungos foram investigados em quatro áreas de Mata Atlântica. Das 18 visitas realizadas nas áreas (oito na Reserva Biológica de Saltinho, quatro no Parque Estadual Dois Irmãos, quatro no Jardim Botânico de Recife e duas na RPPN Frei Caneca) entre 2011 e 2012, foram coletados 54 espécimes de Agaricomycetes terrícolas. Além das coletas, foram revisadas 105 exsicatas depositadas do Herbário URM, resultando em um total de 159 espécimes. Foram identificadas 54 espécies distribuídas em 19 gêneros e 10 famílias. Foram encontrados 12 novos registros para a Região Nordeste e um para o bioma Mata Atlântica. Dos 54 espécimes coletados, foram obtidos sete isolados em meio de cultura, distribuídos em quatro gêneros e cinco espécies. Esses isolados foram testados para celulase, amilase, protease, lacase e peroxidase. Todas as cinco espécies foram negativas para a produção de celulase, amilase e protease e positivas para lacase e peroxidase. Em relação aos aspectos ecológicos, verificou-se que o número de espécimes e espécies coletados foi significativamente maior no período chuvoso. Dos 54 espécimes coletados e dos sete isolados, foram obtidas 17 sequencias da região ITS: cinco sequencias de Amauroderma schomburgkii, duas de A. praetervisum, uma de A. sprucei, uma de Clavulina amazonensis, uma de Clavulinopsis aff. flavella, uma de Clavariaceae, três de Hymenochaete damicornis, uma de Lachnocladium schweinfurthianum, uma de Podoscypha sp1, uma de Ramaria aff. tubulosa e duas de Podoscypha cf. tomentipes (substrato de madeira).

Made available in DSpace on 2014-06-12T15:06:42Z (GMT). No. of bitstreams: 2 arquivo538_1.pdf: 946388 bytes, checksum: eea987472b30d4da6aca6b15f8f6c47d (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Os fungos poróides são caracterizados pelos basidiomas com himenóforo tubular/poróide. A maioria das espécies desse grupo se desenvolve sobre madeira morta decompondo este substrato. Com o objetivo de contribuir com o conhecimento sobre esses organismos no Brasil, a diversidade e aspectos ecológicos de fungos poróides foram investigados em quatro remanescentes de Mata Atlântica em Pernambuco. Percorreu-se um transecto de 500 × 20 m, em nove visitas a campo em cada área de estudo no período de junho/2008 a maio/2009. Foram coletados 922 espécimes correspondendo a 67 espécies, 35 gêneros, oito famílias e duas ordens. Duas espécies são novas para a ciência, cinco são novas ocorrências para o Brasil, dez para o Nordeste e dez para Pernambuco. Nove espécies foram encontradas em todas as áreas, enquanto o total de espécies encontradas em somente uma das quatro áreas foi 39. A diversidade de fungos poróides não diferiu significativamente entre os tipos vegetacionais estudados, enquanto houve diferença entre as épocas chuvosa e seca. O número de ocorrências foi maior do que o esperado no estágio intermediário de decomposição do substrato, e menor do que o esperado no estágio avançado. O número de troncos disponíveis em uma classe de diâmetro parece ser mais importante para a ocorrência de fungos poróides do que a área de madeira disponível nesta classe, enquanto a área parece ser importante para o número de espécies

Cacao (<i>Theobroma cacao </i>L.) is an ancestrally cultivated crop that has been the source of one of the most beloved commodities, chocolate. Its worldwide demand has shaped the history of its cultivation. In Chapter 1, the center of origin of cacao, its center of domestication, the most outstanding movements of germplasm from the Pre-Columbian to the Republican era, the appearance and discovery of major diseases, among other important economic, agricultural and social aspects regarding cacao cultivation are reviewed. The following chapters focus on one of the major pathogens of cacao in the Americas, <i>Moniliophthora roreri</i> causing frosty pod rot disease. Chapter 2 presents evidence that the center of origin of <i>M. roreri </i>is not limited to the Magdalena Valley in Colombia, as other studies have suggested, but extends to Ecuador and the Peruvian Upper Amazon. Chapter 3 focuses on the <i>A </i>and <i>B </i>mating type loci diversity of <i>M. roreri </i>and reports a new <i>A </i>mating allele in Colombia and new mating types in Colombia, Ecuador and infers the presence of even more mating types in Ecuador and the Peruvian Upper Amazon. Additionally, Chapter 3 introduces rapid approaches to collect <i>M. roreri </i> and to diagnose mating types. Finally, Chapter 4 touches the genomic aspect of <i>M. roreri </i>and its close relatives within the Marasmiineae suborder. It presents the most complete genome of a <i>Moniliophthora </i>species generated so far and describes the evolution of predicted effectors and other proteins that might be involved in pathogenicity in this suborder. It also releases a custom program called SyLOCAL that evaluates synteny of a cluster of genes between two genomes.

by Lam Heung-wah, Angora. / Thesis (M. Phil.)--Chinese University of Hong Kong, 1992. / Includes bibliographical references (leaves 124-131). / ACKNOWLEDGEMENTS --- p.iv / ABSTRACT --- p.v / LIST OF ABBREVIATIONS --- p.vii / LIST OF TABLES --- p.viii / LIST OF FIGURES --- p.ix / INTRODUCTION --- p.1 / LITERATURE REVIEW --- p.4 / Chapter A. --- Some Aspects of Cardiovascular Physiology and Pharmacology --- p.4 / Chapter I. --- Fundamental Principles Governing regulation of Arterial Pressure --- p.4 / Chapter II. --- Hypertension --- p.12 / Chapter III. --- Antihypertensive Substances --- p.16 / Chapter B. --- Mushrooms and Their Medicinal Values --- p.29 / Chapter I. --- "The Straw Mushroom, V. volvacea" --- p.29 / Chapter II. --- "Mushrooms, Blood Pressure, and Related Changes" --- p.32 / MATERIALS AND METHODS --- p.39 / Chapter A. --- Basic Preparative Procedures --- p.39 / Chapter I. --- Preparation of Straw Mushroom Extract (SME) --- p.39 / Chapter II. --- Purification of SME by Dialysis --- p.40 / Chapter III. --- Preparation for In vivo Blood Pressure Measurement in Rats --- p.40 / Chapter IV. --- preparation of Right Atrium for In vitro Studies --- p.41 / Chapter V. --- Preparation of Artery Strip for In vitro Studies --- p.42 / Chapter B. --- Experiments Done --- p.43 / Chapter Experiment 1. --- Toxicity of SME --- p.43 / Chapter Experiment 2. --- Hypotensive Effect of SME and Dialyzed Samples --- p.44 / Chapter Experiment 3. --- Pharmacological Antagonist Studies --- p.45 / Chapter Experiment 4. --- Effect of Autonomic Ganglion Blocker and Alpha Blocker on Hypotensive Changes Induced by SME --- p.47 / Chapter Experiment 5. --- Study on Renin-Angiotensin and Kinin Systems --- p.48 / Chapter Experiment 6. --- Urinary and Sodium Excretion in Water-loaded Rats --- p.48 / Chapter Experiment 7. --- Chronotropic and Inotropic Studies on Isolated Right Atria --- p.49 / Chapter Experiment 8. --- Contractile Responses of SME & Its Dialyzed Samples on Rat Tail Artery Strips --- p.50 / Chapter Experiment 9. --- Effect of Adrenergic Blockers in SME Preconstricted Strips --- p.51 / Chapter Experiment 10. --- Acute Oral Effect of DUL8000 and DLL8000 from SME on Blood Pressure --- p.51 / Chapter Experiment 11. --- "Chronic Oral Effect of SME on Blood Pressure, Total Free Cholesterol and Triglyceride Levels" --- p.52 / Chapter C. --- Statistics --- p.55 / RESULTS --- p.56 / Chapter A. --- Toxicity of Straw Mushroom Extract (SME) --- p.56 / Chapter B. --- Effects of SME in Normotensive Rats --- p.56 / Chapter I. --- Blood Pressure Changes --- p.56 / Chapter II. --- Pharmacological Antagonists Studies --- p.58 / Chapter III. --- Converting Enzyme Activity --- p.60 / Chapter IV. --- Urinary and Sodium Excretion --- p.60 / Chapter V. --- In vitro Arterial and Cardiac Effects --- p.61 / Chapter C. --- Cardiovascular Effects of Dialyzed Samples of SME (Molecular Mass cutoffl2000) in Spontaneously Hypertensive Rats (SHR) and Normotensive Rats --- p.61 / Chapter I. --- Blood Pressure Changes --- p.61 / Chapter II. --- In vitro Arterial and Cardiac Effects --- p.62 / Chapter D. --- Cardiovascular Effects of Dialyzed Samples (DUL8000 and DLL8000) in Spontaneously Hypertensive Rats (SHR) and Normotensive Rats --- p.63 / Chapter I. --- Blood Pressure Changes --- p.63 / Chapter II. --- In vitro Arterial and Cardiac Effects --- p.65 / Chapter E. --- The Acute Oral Effect of SME on Blood Pressure --- p.68 / Chapter F. --- "Chronic Dietary Effect of SME on Blood Pressure, Total Free Serum Cholesterol and Triglyceride Levels" --- p.68 / DISCUSSION --- p.109 / Chapter A. --- The Hypotensive Effect of SME --- p.109 / Chapter B. --- The Hypotensive Action of SME: Mechanism of Action --- p.110 / Chapter C. --- The Cardiovascular Active Fractions in SME --- p.113 / Chapter D. --- "The Cardiovascular Effect of DUL8000 and DLL8000 in Rats with Reference to Age, Sex and Strains" --- p.115 / Chapter E. --- The Oral Effect of SME on Blood Pressure and on Serum Cholesterol and Triglyceride Levels --- p.118 / SUMMARY --- p.121 / REFERENCES --- p.124 / APPENDIXES --- p.132

by Yijin Cai. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 185-207). / Introduction / Chapter 1.1 --- Biochemistry of cellulose degradation --- p.1 / Chapter 1.1.1 --- "Occurrence, distribution and structure of cellulose" --- p.1 / Chapter 1.1.2 --- Cellulose-degrading microorganisms --- p.3 / Chapter 1.1.2.1 --- Cellulolytic bacteria --- p.4 / Chapter 1.1.2.2 --- Cellulolytic fungi --- p.4 / Chapter 1.1.3 --- An overview of fungal cellulases --- p.4 / Chapter 1.1.3.1 --- Endoglucanase (EG) --- p.7 / Chapter 1.1.3.2 --- Cellobiohydrolase (CBH) --- p.16 / Chapter 1.1.3.3 --- β-Glucosidase (BGL) --- p.25 / Chapter 1.1.4 --- Synergism between the different components of the cellulolytic systems of filamentous fungi --- p.28 / Chapter 1.1.5 --- Molecular genetics of cellulases --- p.31 / Chapter 1.2 --- Secretion of cellulases by filamentous fungi --- p.32 / Chapter 1.2.1. --- Overview of enzyme secretion in filamentous fungi --- p.33 / Chapter 1.2.2 --- Glycosylation --- p.35 / Chapter 1.2.3 --- Protein secretion and the fungal cell wall --- p.37 / Chapter 1.2.4 --- Factors affecting protein secretion --- p.38 / Chapter 1.3 --- Volvariella volvacea --- p.39 / Chapter 1.4 --- Project aims --- p.43 / Materials and methods / Chapter 2.1 --- Organisms and culture conditions --- p.44 / Chapter 2.1.1 --- Basal medium --- p.44 / Chapter 2.1.2 --- Culture conditions for biomass and enzyme production on different carbon sources --- p.45 / Chapter 2.1.3 --- Culture conditions for large-scale enzyme production for purification --- p.45 / Chapter 2.1.4 --- Culture conditions for confocal microscopy --- p.46 / Chapter 2.1.5 --- Culture conditions for electron microscopy --- p.47 / Chapter 2.2 --- Mycelial extracts --- p.47 / Chapter 2.2.1 --- Large scale extraction --- p.47 / Chapter 2.2.2 --- Small scale extraction --- p.48 / Chapter 2.3 --- Enzyme purification --- p.48 / Chapter 2.3.1 --- Cell-associated enzymes (β-glucosidases) --- p.48 / Chapter 2.3.2 --- Extracellular enzymes --- p.50 / Chapter 2.3.2.1 --- Purification of cellulase complex --- p.50 / Chapter 2.3.2.2 --- Purification of CBH --- p.51 / Chapter 2.3.2.3 --- Purification of endoglucanase-III --- p.53 / Chapter 2.3.2.4 --- Partial purification of β-glucosidases --- p.55 / Chapter 2.3.3 --- Other purification methods --- p.56 / Chapter 2.3.3.1 --- FPLC Phenylsuperose hydrophobic interaction chromatography --- p.56 / Chapter 2.3.3.2 --- Affinity gel chromatography --- p.56 / Chapter 2.3.3.3 --- Isoelectric focusing by Rotorfor --- p.57 / Chapter 2.3.3.4 --- Preparative gel electrophoresis --- p.57 / Chapter 2.3.3.5 --- (NH4)2S04 Precipitation --- p.58 / Chapter 2.4 --- Electrophoresis --- p.59 / Chapter 2.4.1 --- Mini Protean-II system (BioRad) --- p.59 / Chapter 2.4.2 --- PhastGel system (Pharmacia) --- p.60 / Chapter 2.5 --- Enzyme assays --- p.61 / Chapter 2.5.1 --- β-Glucosidase --- p.61 / Chapter 2.5.2 --- Endoglucanase --- p.63 / Chapter 2.5.3 --- Cellobiohydrolase --- p.65 / Chapter 2.6 --- β-Glucosidase characterization studies --- p.66 / Chapter 2.6.1 --- pH optimum --- p.66 / Chapter 2.6.2 --- Temperature optimum --- p.66 / Chapter 2.6.3 --- Thermal stability --- p.66 / Chapter 2.6.4 --- Kinetic parameters --- p.67 / Chapter 2.6.5 --- Enzyme inhibitor studies --- p.67 / Chapter 2.6.6 --- Effect of lignin-derived phenolic monomers --- p.67 / Chapter 2.6.7 --- Substrate specificity towards p-nitrophenyl-linked glycosides --- p.67 / Chapter 2.6.8 --- "Substrate specificity towards different cellulosic substrates, mono- and disaccharides, hemicellulose, sugar alcohols and saponins" --- p.68 / Chapter 2.6.9 --- Cellulose-binding assay --- p.68 / Chapter 2.6.10 --- Effect of purified β-glucosidase on the production of glucose from crystalline cellulose and carboxymethylcellulose by Aspergillus niger cellulase --- p.69 / Chapter 2.7 --- Miscellaneous analytical methods --- p.69 / Chapter 2.7.1 --- Protein determination --- p.69 / Chapter 2.7.2 --- Determination of isoelectric points --- p.69 / Chapter 2.7.3 --- Activity staining of gels for cellulolytic enzyme activity --- p.70 / Chapter 2.7.4 --- Staining for glycoprotein --- p.71 / Chapter 2.7.5 --- Molecular weight determination --- p.71 / Chapter 2.8 --- "Production, purification and specificity of antibodies to β-glucosidases and EG-III" --- p.72 / Chapter 2.8.1 --- Antibodies to β-glucosidases --- p.72 / Chapter 2.8.2 --- Antibodies to EG-III --- p.74 / Chapter 2.9 --- Immunocytochemical studies --- p.75 / Chapter 2.9.1 --- Confocal laser scanning microscopy --- p.75 / Chapter 2.9.1.1 --- β-Glucosidases --- p.75 / Chapter 2.9.1.2 --- Endoglucanase-III --- p.75 / Chapter 2.9.2 --- Transmission electron microscopy --- p.76 / Chapter 2.9.3 --- Scanning electron microscopy --- p.77 / Chapter 2.10 --- Chemicals --- p.77 / Results / Chapter 3.1 --- "Effect of culture conditions on the growth of, and the production of cellulolytic enzymes, by V. volvacea" --- p.79 / Chapter 3.1.1 --- Growth --- p.79 / Chapter 3.1.2 --- Endoglucanases --- p.81 / Chapter 3.1.3 --- Cellobiohydrolase --- p.84 / Chapter 3.1.4 --- β-Glucosidase --- p.87 / Chapter 3.2 --- Purification of cellulolytic enzymes from V. volvacea --- p.92 / Chapter 3.2.1 --- Preliminary purification of V. volvacea extracellular cellulolytic enzymes --- p.92 / Chapter 3.2.1.1. --- (NH4)2SO4 precipitation --- p.92 / Chapter 3.2.1.2 --- Ultrafiltration --- p.94 / Chapter 3.2.1.3 --- Batch adsorption by anion exchanger --- p.94 / Chapter 3.2.1.4 --- Separation by column chromatography --- p.96 / Chapter 3.2.2 --- CBH enzymes …… --- p.102 / Chapter 3.2.3 --- Endoglucanase enzymes --- p.106 / Chapter 3.2.4 --- Cell-associated β-glucosidase enzymes --- p.113 / Chapter 3.2.5 --- Extracellular β-glucosidase enzymes --- p.120 / Chapter 3.3 --- Characterization of cell-associated β-glucosidases from V. volvacea --- p.122 / Chapter 3.3.1 --- Influence of pH and temperature --- p.122 / Chapter 3.3.2 --- Enzyme stability --- p.125 / Chapter 3.3.3 --- Kinetic parameters --- p.128 / Chapter 3.3.4 --- Enzyme inhibitors --- p.131 / Chapter 3.3.5 --- Substrate specificity --- p.137 / Chapter 3.3.6 --- Cellulose-binding and hydrolysing properties --- p.139 / Chapter 3.3.7 --- Molecular weights and isoelectric points --- p.142 / Chapter 3.4 --- Immunocytochemical studies on cellulolytic enzymes from V. volvacea --- p.146 / Chapter 3.4.1 --- Cell-associated β-glucosidase --- p.146 / Chapter 3.4.1.1 --- "Production, specificity and purification of polyclonal antibody" --- p.146 / Chapter 3.4.1.2. --- Localization --- p.151 / Chapter 3.4.1.3. --- Localization of cell-associated β-glucosidases by immuno- labelling --- p.151 / Chapter 3.4.2 --- Endoglucanase-III --- p.161 / Discussion / Chapter 4.1 --- Production --- p.163 / Chapter 4.2 --- Composition of cellulolytic enzyme system of V. volvacea --- p.168 / Chapter 4.3 --- Properties --- p.172 / Chapter 4.4 --- Localization --- p.179 / References --- p.185