Perfil de secreção de citocinas e de ativação de células endoteliais após interação com cepas selvagens e mutantes de Aspergillus fumigatus

Apesar do desenvolvimento de novas drogas antifúngicas e da sua utilização como terapia profilática visando à prevenção de infecções fúngicas invasivas, estas ainda constituem-se num problema emergente, com elevadas taxas de mortalidade. Neste contexto, destaca-se a aspergilose invasiva, uma infecção fúngica oportunista que acomete pacientes com neutropenia profunda e prolongada, principalmente os pacientes com leucemia aguda ou submetidos a transplante de medula óssea. Aspergillus fumigatus, um fungo filamentoso, é o principal agente etiológico da aspergilose invasiva, sendo um patógeno angioinvasivo. As hifas deste fungo são capazes de causar injúria e ativação endotelial, induzindo o endotélio a um fenótipo pró-trombótico, que por sua vez, é mediado pela secreção de citocinas pró-inflamatórias, em especial, o TNF-α. O presente trabalho teve como objetivo estudar a capacidade de cepas mutantes de A. fumigatus em ativar células endoteliais, avaliando o perfil de secreção de citocinas em meio condicionado e a expressão de fator tecidual. Resumidamente, monocamadas confluentes de células endoteliais isoladas da veia umbilical humana foram incubadas com conídios e tubos germinativos de cepas selvagens (Af293 e Ku80) e mutantes (Δugm1, ΔcalA, ΔcrzA, ΔprtT) de A. fumigatus. A taxa de adesão e endocitose destas cepas às monocamadas de HUVEC foi avaliada a partir de um ensaio quantitativo de imunofluorescência diferencial. O perfil cinético de secreção de citocinas foi determinado em meio condicionado das HUVECs, por ensaio de multiplex para IL-6, IL-8 e TNF-α. A ativação endotelial, por sua vez, foi determinada pela expressão de fator tecidual por RT-PCR em tempo real. Os resultados obtidos demonstraram que a mutante para o gene ugm1, responsável por codificar a enzima UDP-galactopiranose mutase, que converte resíduos de galactopiranose a galactofuranose, apresentou um fenótipo hiperaderente às células endoteliais e um estímulo 10 vezes maior à secreção de TNF-α e 2,5 vezes maior a secreção de IL-6, quando comparada a ativação observada para as cepas selvagens. A galactofuranose é um componente importante de glicoconjugados da parede celular de A. fumigatus. Dessa forma, a ausência desse monossacarídeo na célula fúngica leva a um mecanismo compensatório caracterizado por um aumento na expressão de moléculas de galactosaminogalactana na parede celular. De maneira contrária, mutantes para os genes calA e crzA, apresentaram um fenótipo hipoaderente às HUVECs e uma perda na capacidade de induzir a secreção de citocinas e ativar o endotélio. Essas mutantes apresentam deleções que interferem na via de cálcio/calcineurina, responsável por regular a morfogênese e virulência de A. fumigatus, além de apresentarem alterações no conteúdo de beta-1-3 glucana. Já a cepa ΔprtT, mutante para o fator de transcrição prtT que regula a secreção de múltiplas proteases, apresentou um fenótipo de adesão, estímulo e ativação endotelial semelhante ao observado para as cepas selvagens. A comparação entre a capacidade de conídios e tubos germinativos em ativar células endoteliais, corroborou achados anteriores da literatura que reportam que só hifas são capazes de ativar células endoteliais, independentemente da sua viabilidade. Os dados deste estudo permitiram concluir que dentre os componentes de superfície celular de A. fumigatus, os polímeros de galactose, em especial a galactosaminogalactana, parecem ser responsáveis, pelo menos em parte, pelos mecanismos de interação e ativação endotelial.

Coping with the Lionfish Invasion: can targeted removals yield beneficial effects?

Invasive species generate significant environmental and economic costs, with maintenance management constituting a major expenditure. Such costs are generated by invasive Indo-Pacific lionfish (Pterois spp.) that further threaten already stressed coral reefs in the western Atlantic Ocean and Caribbean Sea. This brief review documents rapid range expansion and potential impacts of lionfish. In addition, preliminary experimental data from targeted removals contribute to debates about maintenance management. Removals at sites off Little Cayman Island shifted the size frequency distribution of remaining lionfish toward smaller individuals whose stomachs contained less prey and fewer fish. Fewer lionfish and decreased predation on threatened grouper, herbivores and other economically and ecologically important fishes represent key steps toward protecting reefs. However, complete evaluation of success requires long-term data detailing immigration and recruitment by lionfish, compensatory growth and reproduction of lionfish, reduced direct effects on prey assemblages, and reduced indirect effects mediated by competition for food. Preventing introductions is the best way to avoid impacts from invasive species and early detection linked to rapid response ranks second. Nevertheless, results from this case study suggest that targeted removals represent a viable option for shifting direct impacts of invasive lionfish away from highly vulnerable components of ecosystems.

A tree-ring perspective on the extreme wetness of the early 1900’s in the western United States [abstract]

Ring-width indices from 136 sites in the area from northern Montana to southern New Mexico between latitudes 103°W and 111°W were examined to infer periods of anomalous wetness for the years 1700-1964. Sites were grouped into north, central and south regions, and the gross regional tree-ring fluctuations were compared. The results indicate that the period 1905-1917 was unique in the 265-year record for the combined magnitude, duration, and north/south coherence of the growth anomaly of much lesser magnitude occurred in the 1830's-1840's [sic]. Both this and the 1905-1917 anomaly appear from time-series plots to be manifestations of low-frequency growth variations at wave lengths between about 20 and 60 years.

A biogeographic assessment of the Channel Islands National Marine Sanctuary: a review of boundary expansion concepts for NOAA’s National Marine Sanctuary Program

The priority management goal of the National Marine Sanctuaries Program (NMSP) is to protect marine ecosystems and biodiversity. This goal requires an understanding of broad-scale ecological relationships and linkages between marine resources and physical oceanography to support an ecosystem management approach. The Channel Islands National Marine Sanctuary (CINMS) is currently reviewing its management plan and investigating boundary expansion. A management plan study area (henceforth, Study Area) was described that extends from the current boundary north to the mainland, and extends north to Point Sal and south to Point Dume. Six additional boundary concepts were developed that vary in area and include the majority of the Study Area. The NMSP and CINMS partnered with NOAA’s National Centers for Coastal Ocean Science Biogeography Team to conduct a biogeographic assessment to characterize marine resources and oceanographic patterns within and adjacent to the sanctuary. This assessment includes a suite of quantitative spatial and statistical analyses that characterize biological and oceanographic patterns in the marine region from Point Sal to the U.S.-Mexico border. These data were analyzed using an index which evaluates an ecological “cost-benefit” within the proposed boundary concepts and the Study Area. The sanctuary resides in a dynamic setting where two oceanographic regimes meet. Cold northern waters mix with warm southern waters around the Channel Islands creating an area of transition that strongly influences the regions oceanography. In turn, these processes drive the biological distributions within the region. This assessment analyzes bathymetry, benthic substrate, bathymetric life-zones, sea surface temperature, primary production, currents, submerged aquatic vegetation, and kelp in the context of broad-scale patterns and relative to the proposed boundary concepts and the Study Area. Boundary cost-benefit results for these parameters were variable due to their dynamic nature; however, when analyzed in composite the Study Area and Boundary Concept 2 were considered the most favorable. Biological data were collected from numerous resource agencies and university scientists for this assessment. Fish and invertebrate trawl data were used to characterize community structure. Habitat suitability models were developed for 15 species of macroinvertebrates and 11 species of fish that have significant ecological, commercial, or recreational importance in the region and general patterns of ichthyoplankton distribution are described. Six surveys of ship and plane at-sea surveys were used to model marine bird diversity from Point Arena to the U.S.-Mexico border. Additional surveys were utilized to estimate density and colony counts for nine bird species. Critical habitat for western snowy plover and the location of California least tern breeding pairs were also analyzed. At-sea surveys were also used to describe the distribution of 14 species of cetaceans and five species of pinnipeds. Boundary concept cost-benefit indices revealed that Boundary Concept 2 and the Study Area were most favorable for the majority of the species-specific analyses. Boundary Concept 3 was most favorable for bird diversity across the region. Inadequate spatial resolution for fish and invertebrate community data and incompatible sampling effort information for bird and mammal data precluded boundary cost-benefit analysis.