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During nocturnal field expeditions in the Brazilian Atlantic Rainforest, an unexpected bioluminescent fungus with reduced form was found. Based on morphological data, the taxon was first identified as belonging to the cyphelloid genus Maireina, but in our phylogenetic analyses, Maireina was recovered and confirmed as a paraphyletic group related to genera Merismodes and Cyphellopsis. Maireina filipendula, Ma. monacha, and Ma. subsphaerospora are herein transferred to Merismodes. Based upon morphological and molecular characters, the bioluminescent cyphelloid taxon is described as the new genus Eoscyphella, characterized by a vasiform to urceolate basidiomata, subglobose to broadly ellipsoid basidiospores, being pigmented, weakly to densely encrusted external hyphae, regularly bi-spored basidia, unclamped hyphae, and an absence of both conspicuous long external hairs and hymenial cystidia. Phylogenetic analyses based on ITS rDNA and LSU rDNA support the proposal of the new genus and confirm its position in Cyphellopsidaceae. Eoscyphella luciurceolata represents a new lineage of bioluminescent basidiomycetes with reduced forms.
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In the original publication [...].
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Gymnopilus consists in a widely distributed genus of mushroom-forming fungi, especially in tropical regions of the world. Literature on Gymnopilus representatives reports the presence of oligoisoprenoids, and styrylpyrones. Considering the large number of secondary metabolites that basidiomycetes might contain, dereplication tools such as GNPS (Global Natural Products Social Molecular Networking), has become important in prospecting metabolites, saving time and work on isolation and characterization of natural products. Thus, this work identified the wild mushroom Gymnopilus imperialis and dereplicated their extracts with the aid of GNPS to annotate oligoisoprenoids. It was possible to annotate 24 oligoisoprenoids from methanol, dichloromethaneand ethyl acetate extracts of G. imperialis, 4 of them from GNPS spectral library match, and 20 from prediction based on molecular network. Moreover HRMS-ESI-(+) dereplication of the acetate extract annotated bisnoryangonin and hispidin. To the best of our knowledge, this is the first report on the annotation of a series of gymnopilins analogues based on GNPS molecular network. Our findings suggest that GNPS might be an effective, rapid, and open-source device to identify compounds and predict analogues.
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Agaricales , Basidiomycota , Basidiomycota/genética , Cromatografia Líquida de Alta Pressão , MetabolômicaRESUMO
The l-DOPA dioxygenase of Amanita muscaria (AmDODA) participates in the biosynthesis of betalain- and hygroaurin-type natural pigments. AmDODA is encoded by the dodA gene, whose DNA sequence was inferred from cDNA and gDNA libraries almost 30 years ago. However, reports on its heterologous expression rely on either the original 5'-truncated cDNA plasmid or artificial gene synthesis. We provide unequivocal evidence that the heterologous expression of AmDODA from A. muscaria specimens is not possible by using the coding sequence previously inferred for dodA. Here, we rectify and reannotate the full-length coding sequence for AmDODA and express a 205-aa His-tagged active enzyme, which was used to produce the l-DOPA hygroaurin, a rare fungal pigment. Moreover, AmDODA and other isozymes from bacteria were submitted to de novo folding using deep learning algorithms, and their putative active sites were inferred and compared. The wide catalytic pocket of AmDODA and the presence of the His-His-His and His-His-Asp motifs can provide insight into the dual cleavage of l-DOPA at positions 2,3 and 4,5 as per the mechanism proposed for nonheme dioxygenases.
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Environmental pollutants are today a major concern and an intensely discussed topic on the global agenda for sustainable development. They include a wide range of organic compounds, such as pharmaceutical waste, pesticides, plastics, and volatile organic compounds that can be found in air, soil, water bodies, sewage, and industrial wastewater. In addition to impacting fauna, flora, and fungi, skin absorption, inhalation, and ingestion of some pollutants can also negatively affect human health. Fungi play a crucial role in the decomposition and cycle of natural and synthetic substances. They exhibit a variety of growth, metabolic, morphological, and reproductive strategies and can be found in association with animals, plants, algae, and cyanobacteria. There are fungal strains that occur naturally in soil, sediment, and water that have inherent abilities to survive with contaminants, making the organism important for bioassay applications. In this context, we reviewed the applications of fungal-based bioassays as a versatile tool for environmental monitoring.
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Gymnopilus consists of a widely distributed genus of basidiomycetes, especially in tropical regions of the world, such as Japan, Australia, Paraguay, and Brazil. This genus biosynthesizes interesting bioactive compounds, such as sesquiterpenoids, oligoisoprenoids, styrylpyrones, and lectins. In the present study, the aqueous extract of the basidiomata of Gymnopilus imperialis (Basidiomycota, Agaricomycetes, Agaricales, Hymenogastraceae) was obtained by using the accelerated solvent extraction (ASE) technique, followed by the precipitation of polysaccharide fraction with ethanol. Further purification by freeze-thawing processes, Fehling solution precipitation, and membrane dialysis with different pore sizes yield three main polysaccharide fractions (Gi-MRSW, Gi-PFME, and Gi-SFME). According to monosaccharide composition and 13C-NMR data, the Gi-MRSW and Gi-SFME fractions showed to be composed mainly of ß-glucans and Gi-PFME by a heterogalactan. Moreover, the immunomodulatory potential of Gi-MRSW was evaluated using RAW 264.7 murine macrophage as a study model. The nitric oxide production was significantly increased in treated samples, and the expression of inducible nitric oxide synthase (iNOS) showed that the fraction Gi-MRSW from G. imperialis induces the M1 polarization phenotype.
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Neonothopanus gardneri, also known as coconut flower mushroom (flor-de-coco), is a Brazilian bioluminescent basidiomycete found in Palm Forest, a transitional biome between the Amazonian Forest and Caatinga (Savanna-like vegetation) in Northeast Brazil, especially in Piauí State. Recent advances toward the elucidation of fungal bioluminescence have contributed to the discovery of four genes (hisps, h3h, luz and cph) involved with the bioluminescence process, the so-called Caffeic Acid Cycle (CAC) and to develop biotechnological applications such autoluminescent tobacco plants and luciferase-based reporter genes. High-yield and -quality RNA-extraction methods are required for most of these purposes. Herein, four methods for RNA isolation from the mycelium of N. gardneri were evaluated: RNeasy® kit (QIAGEN), TRI+, TRI18G+, and TRI26G+. Highest RNA yield was observed for TRI18G+ and TRI26G+ methods, an increase of ~130% in comparison to the RNeasy® method and of ~40% to the TRI+ protocol. All the RNA samples showed good purity and integrity, except by gDNA contamination in RNA samples produced with the RNeasy® method. High quality of RNA samples was confirmed by successful cDNA synthesis and PCR amplification of the coding sequence of h3h gene, responsible for the hydroxylation of the precursor of fungal luciferin (3-hydroxyhispidin). Similarly, RT-qPCR amplification of ef-tu gene, related to the protein biosynthesis in the cell, was demonstrated from RNA samples. This is the first report of a reproducible, time-saving and low-cost optimized method for isolation of high-quality and -yield, DNA-free RNA from a bioluminescent fungus, but that can also be useful for other basidiomycetes.
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Agaricales/genética , Medições Luminescentes/métodos , Micélio/genética , Técnicas de Tipagem Micológica/métodos , RNA Fúngico/isolamento & purificação , Agaricales/isolamento & purificação , Agaricales/metabolismo , Biotecnologia , Brasil , DNA Complementar , Ecossistema , Florestas , Luciferinas , Tipagem Molecular/métodos , Reação em Cadeia da Polimerase , Biossíntese de ProteínasRESUMO
The evolution of broadly attached thoracic lobes could have increased the body temperature excess of ancient wingless insects by 55 percent over that of lobeless forms. The subsequent expansion of these thoracic lobes for behavioral thermoregulation could have provided the morphological stage required for the evolution of functional wings.
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Invasion by the African grass Andropogon gayanus is drastically altering the understory structure of oligotrophic savannas in tropical Australia. We compared nitrogen (N) relations and phenology of A. gayanus and native grasses to examine the impact of invasion on N cycling and to determine possible reasons for invasiveness of A. gayanus. Andropogon gayanus produced up to 10 and four times more shoot phytomass and root biomass, with up to seven and 2.5 times greater shoot and root N pools than native grass understory. These pronounced differences in phytomass and N pools between A. gayanus and native grasses were associated with an altered N cycle. Most growth occurs in the wet season when, compared with native grasses, dominance of A. gayanus was associated with significantly lower total soil N pools, lower nitrification rates, up to three times lower soil nitrate availability, and up to three times higher soil ammonium availability. Uptake kinetics for different N sources were studied with excised roots of three grass species ex situ. Excised roots of A. gayanus had an over six times higher-uptake rate of ammonium than roots of native grasses, while native grass Eriachne triseta had a three times higher uptake rate of nitrate than A. gayanus. We hypothesize that A. gayanus stimulates ammonification but inhibits nitrification, as was shown to occur in its native range in Africa, and that this modification of the soil N cycle is linked to the species' preference for ammonium as an N source. This mechanism could result in altered soil N relations and could enhance the competitive superiority and persistence of A. gayanus in Australian savannas.
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Andropogon/metabolismo , Biomassa , Compostos de Nitrogênio/análise , Nitrogênio/metabolismo , Solo/análise , Andropogon/crescimento & desenvolvimento , Ecossistema , Nitrogênio/análise , Northern Territory , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismoRESUMO
The modification of river flow regimes poses a significant threat to the world's freshwater ecosystems. Northern Australia's freshwater resources, particularly dry season river flows, are being increasingly modified to support human development, potentially threatening aquatic ecosystems and biodiversity, including fish. More information is urgently needed on the ecology of fishes in this region, including their habitat requirements, to support water policy and management to ensure future sustainable development. This study used electrofishing and habitat survey methods to quantify the dry season habitat use of 20 common freshwater fish taxa in the Daly River in Australia's wet-dry tropics. Of twenty measured habitat variables, water depth and velocity were the two most important factors discriminating fish habitat use for the majority of taxa. Four distinct fish habitat guilds were identified, largely classified according to depth, velocity and structural complexity. Ontogenetic shifts in habitat use were also observed in three species. This study highlights the need to maintain dry season river flows that support a diversity of riverine mesohabitats for freshwater fishes. In particular, shallow fast-flowing areas provided critical nursery and refuge habitats for some species, but are vulnerable to water level reductions due to water extraction. By highlighting the importance of a diversity of habitats for fishes, this study assists water managers in future decision making on the ecological risks of water extractions from tropical rivers, and especially the need to maintain dry season low flows to protect the habitats of native fish.
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Peixes/fisiologia , Animais , Austrália , Biodiversidade , Conservação dos Recursos Naturais/métodos , Ecologia , Ecossistema , Água Doce , Rios , Alimentos Marinhos , Estações do Ano , ÁguaRESUMO
The African grass Andropogon gayanus Kunth. is invading Australian savannas, altering their ecological and biogeochemical function. To assess impacts on nitrogen (N) cycling, we quantified litter decomposition and N dynamics of grass litter in native grass and A. gayanus invaded savanna using destructive in situ grass litter harvests and litterbag incubations (soil surface and aerial position). Only 30% of the A. gayanus in situ litter decomposed, compared to 61% of the native grass litter, due to the former being largely comprised of highly resistant A. gayanus stem. In contrast to the stem, A. gayanus leaf decomposition was approximately 3- and 2-times higher than the dominant native grass, Alloteropsis semilata at the surface and aerial position, respectively. Lower initial lignin concentrations, and higher consumption by termites, accounted for the greater surface decomposition rate of A. gayanus. N flux estimates suggest the N release of A. gayanus litter is insufficient to compensate for increased N uptake and N loss via fire in invaded plots. Annually burnt invaded savanna may lose up to 8.2% of the upper soil N pool over a decade. Without additional inputs via biological N fixation, A. gayanus invasion is likely to diminish the N capital of Australia's frequently burnt savannas.
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Andropogon/metabolismo , Espécies Introduzidas , Nitrogênio/metabolismo , Poaceae/metabolismo , Animais , Austrália , Ecossistema , Incêndios , Pradaria , Isópteros , Folhas de Planta/metabolismoRESUMO
Transdisciplinary research, involving close collaboration between researchers and the users of research, has been a feature of environmental problem solving for several decades, often spurred by the need to find negotiated outcomes to intractable problems. In 2005, the Australian government allocated funding to its environment portfolio for public good research, which resulted in consecutive four-year programmes (Commonwealth Environmental Research Facilities, National Environmental Research Program). In April 2014, representatives of the funders, researchers and research users associated with these programmes met to reflect on eight years of experience with these collaborative research models. This structured reflection concluded that successful multi-institutional transdisciplinary research is necessarily a joint enterprise between funding agencies, researchers and the end users of research. The design and governance of research programmes need to explicitly recognise shared accountabilities among the participants, while respecting the different perspectives of each group. Experience shows that traditional incentive systems for academic researchers, current trends in public sector management, and loose organisation of many end users, work against sustained transdisciplinary research on intractable problems, which require continuity and adaptive learning by all three parties. The likelihood of research influencing and improving environmental policy and management is maximised when researchers, funders and research users have shared goals; there is sufficient continuity of personnel to build trust and sustain dialogue throughout the research process from issue scoping to application of findings; and there is sufficient flexibility in the funding, structure and operation of transdisciplinary research initiatives to enable the enterprise to assimilate and respond to new knowledge and situations.
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Conservação dos Recursos Naturais/métodos , Ecologia , Comportamento Cooperativo , PesquisaRESUMO
Methods are described used to link the Community Health Index and the National Health Service Central Register (NHSCR) in Scotland to provide a basis for a national patient index. The linkage used a combination of deterministic and probability matching techniques. A best-link principle was used by which each Community Health Index record was allowed to link only to the NHSCR record with which it achieved the highest match weight. This strategy, applied in the context of two files which each covered virtually the entire population of Scotland, increased the accuracy of linkage approximately a thousand-fold compared with the likely results of a less structured probability matching approach. By this means, 98.8% of linkable records were linked automatically with a sufficient degree of confidence for administrative purposes.
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Planejamento em Saúde Comunitária , Registro Médico Coordenado , Sistema de Registros , Humanos , Probabilidade , EscóciaRESUMO
Prior to 1980, almost all drug advertising was direted to physicians, the parties responsible for prescribing medications to patients. Beginning in 1980, however, drug companies began advertising directly to patients by various means, specifically television and print adversisements and sponsorship of sports and similar events. During the last decade, as computers have become a common household item, the Internet is now another source of advertising for drug manufacturers. This change in advertising has been effective, since research has demonstrated that physicians' prescribing decisions are, in fact, influenced heavily by patients' requests for specific medications. In 1997, the US Food and Drug Administration devised specific guidelines for the television advertising of drugs produced by pharmaceutical manufacturers. Since youth and beauty have long been valued feminine qualities in the US, where menopausal women are assumed to suffer most from the effects of aging, pharmaceutical companies targeted women for the promotion of hormone-containing drugs that were purported to minimize the effects of menopause and ensure better health. As a result, advertising has played a key role in the prescribing of hormone replacement therapy in the US for the last 60 years. Since many of the advertised benefits of hormone replacement therapy are never realized by the patient, and the hazards of treatment are often minimized, it remains the responsibility of clinicians to safeguard their patients' health by basing the decision to prescribe on valid scientific evidence rather than the message of an advertising pitch.
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Social competence assessment and training has long focused on specific skills within the clinical setting. In addition, emphasis has been placed on identifying deficits relative to an arbitrary, often idiosyncratic metric. In this article, we discuss the importance of the principles that underlie communication and which are reflected in the range of behaviours described as 'social competence'. We review methods we have found productive in the training of these principles with persons who have suffered traumatic brain injuries.