RESUMO
Soil pollutants such as hydrocarbons can induce toxic effects in plants and associated arbuscular mycorrhizal fungi (AMF). This study was conducted to evaluate if the legume Lotus corniculatus and the grass Elymus trachycaulus and arbuscular mycorrhizal fungi could grow in two oil sands processing by-products after bitumen extraction from the oil sands in northern Alberta, Canada. Substrate treatments were coarse tailings sand (CTS), a mix of dry mature fine tailings (MFT) with CTS (1:1) and Pleistocene sandy soil (hydrocarbon free); microbial treatments were without AMF, with AMF and AMF plus soil bacteria isolated from oil sands reclamation sites. Plant biomass, root morphology, leaf water content, shoot tissue phosphorus content and mycorrhizal colonization were evaluated. Both plant species had reduced growth in CTS and tailings mix relative to sandy soil. AMF frequency and intensity in roots of E. trachycaulus was not influenced by soil hydrocarbons; however, it decreased significantly over time in roots of L. corniculatus without bacteria in CTS. Mycorrhizal inoculation alone did not significantly improve plant growth in CTS and tailings mix; however, inoculation with mycorrhizae plus bacteria led to a significantly positive response of both plant species in CTS. Thus, combined inoculation with selected mycorrhizae and bacteria led to synergistic effects. Such combinations may be used in future to improve plant growth in reclamation of CTS and tailings mix.
Assuntos
Elymus/crescimento & desenvolvimento , Elymus/microbiologia , Micorrizas , Campos de Petróleo e Gás , Poluição por Petróleo , Alberta , Bactérias , Biomassa , Hidrocarbonetos , Raízes de Plantas/microbiologia , Solo , Poluentes do SoloRESUMO
BACKGROUND: Recent clinical studies show that tyrosine kinase inhibitors slow the rate of lung function decline and decrease the number of acute exacerbations in patients with Idiopathic Pulmonary Fibrosis (IPF). However, in the murine bleomycin model of fibrosis, not all tyrosine kinase signaling is detrimental. Exogenous ligands Fibroblast Growth Factor (FGF) 7 and 10 improve murine lung repair and increase survival after injury via tyrosine kinase FGF receptor 2b-signaling. Therefore, the level and location of FGF/FGFR expression as well as the exogenous effect of the most highly expressed FGFR2b ligand, FGF1, was analyzed on human lung fibroblasts. METHODS: FGF ligand and receptor expression was evaluated in donor and IPF whole lung homogenates using western blotting and qPCR. Immunohistochemistry for FGF1 and FGFR1/2/3/4 were performed on human lung tissue. Lastly, the effects of FGF1, a potent, multi-FGFR ligand, were studied on primary cultures of IPF and non-IPF donor fibroblasts. Western blots for pro-fibrotic markers, proliferation, FACS for apoptosis, transwell assays and MetaMorph analyses on cell cultures were performed. RESULTS: Whole lung homogenate analyses revealed decreased FGFR b-isoform expression, and an increase in FGFR c-isoform expression. Of the FGFR2b-ligands, FGF1 was the most significantly increased in IPF patients; downstream targets of FGF-signaling, p-ERK1/2 and p-AKT were also increased. Immunohistochemistry revealed FGF1 co-localization within basal cell sheets, myofibroblast foci, and Surfactant protein-C positive alveolar epithelial type-II cells as well as co-localization with FGFR1, FGFR2, FGFR3, FGFR4 and myofibroblasts expressing the migratory marker Fascin. Both alone and in the presence of heparin, FGF1 led to increased MAPK-signaling in primary lung fibroblasts. While smooth muscle actin was unchanged, heparin + FGF1 decreased collagen production in IPF fibroblasts. In addition, FGF1 + heparin increased apoptosis and cell migration. The FGFR inhibitor (PD173074) attenuated these effects. CONCLUSIONS: Strong expression of FGF1/FGFRs in pathogenic regions of IPF suggest that aberrant FGF1-FGFR signaling is increased in IPF patients and may contribute to the pathogenesis of lung fibrosis by supporting fibroblast migration and increased MAPK-signaling.
Assuntos
Fator 1 de Crescimento de Fibroblastos/biossíntese , Fibrose Pulmonar Idiopática/metabolismo , Receptores de Fatores de Crescimento de Fibroblastos/biossíntese , Movimento Celular/fisiologia , Células Cultivadas , Regulação da Expressão Gênica , Humanos , Fibrose Pulmonar Idiopática/patologia , Pulmão/metabolismo , Pulmão/patologiaRESUMO
Recultivation of disturbed oil sand mining areas is an issue of increasing importance. Nevertheless only little is known about the fate of organic matter, cell abundances and microbial community structures during oil sand processing, tailings management and initial soil development on reclamation sites. Thus the focus of this work is on biogeochemical changes of mined oil sands through the entire process chain until its use as substratum for newly developing soils on reclamation sites. Therefore, oil sand, mature fine tailings (MFTs) from tailings ponds and drying cells and tailings sand covered with peat-mineral mix (PMM) as part of land reclamation were analyzed. The sample set was selected to address the question whether changes in the above-mentioned biogeochemical parameters can be related to oil sand processing or biological processes and how these changes influence microbial activities and soil development. GC-MS analyses of oil-derived biomarkers reveal that these compounds remain unaffected by oil sand processing and biological activity. In contrast, changes in polycyclic aromatic hydrocarbon (PAH) abundance and pattern can be observed along the process chain. Especially naphthalenes, phenanthrenes and chrysenes are altered or absent on reclamation sites. Furthermore, root-bearing horizons on reclamation sites exhibit cell abundances at least ten times higher (10(8) to 10(9) cells g(-1)) than in oil sand and MFT samples (10(7) cells g(-1)) and show a higher diversity in their microbial community structure. Nitrate in the pore water and roots derived from the PMM seem to be the most important stimulants for microbial growth. The combined data show that the observed compositional changes are mostly related to biological activity and the addition of exogenous organic components (PMM), whereas oil extraction, tailings dewatering and compaction do not have significant influences on the evaluated compounds. Microbial community composition remains relatively stable through the entire process chain.
Assuntos
Consórcios Microbianos , Mineração , Campos de Petróleo e Gás/microbiologia , Resíduos Industriais/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Químicos da Água/análiseRESUMO
Monoclonal antibody (MAb) 2c, specific for glycoprotein B of herpes simplex virus (HSV), had been shown to mediate clearance of infection from the mucous membranes of mice, thereby completely inhibiting mucocutaneous inflammation and lethality, even in mice depleted of both CD4(+) and CD8(+) cells. Additionally, ganglionic infection was highly restricted. In vitro, MAb 2c exhibits a potent complement-independent neutralising activity against HSV type 1 and 2, completely inhibits the viral cell-to-cell spread as well as the syncytium formation induced by syncytial HSV strains (Eis-Hübinger et al. in Intervirology 32:351-360, 1991; Eis-Hübinger et al. in J Gen Virol 74:379-385, 1993). Here, we describe the mapping of the epitope for MAb 2c. The antibody was found to recognise a discontinuous epitope comprised of the HSV type 1 glycoprotein B residues 299 to 305 and one or more additional discontinuous regions that can be mimicked by the sequence FEDF. Identification of the epitope was confirmed by loss of antibody binding to mutated glycoprotein B with replacement of the epitopic key residues, expressed in COS-1 cells. Similarly, MAb 2c was not able to neutralise HSV mutants with altered key residues, and MAb 2c was ineffective in mice inoculated with such mutants. Interestingly, identification and fine-mapping of the discontinuous epitope was not achieved by binding studies with truncated glycoprotein B variants expressed in COS cells but by peptide scanning with synthetic overlapping peptides and peptide key motif analysis. Reactivity of MAb 2c was immensely increased towards a peptide composed of the glycoprotein B residues 299 to 305, a glycine linker, and a C-terminal FEDF motif. If it could be demonstrated that antibodies of the specificity and bioactivity of MAb 2c can be induced by the epitope or a peptide mimicking the epitope, strategies for active immunisation might be conceivable.
Assuntos
Anticorpos Monoclonais/imunologia , Mapeamento de Epitopos , Herpes Simples/prevenção & controle , Herpesvirus Humano 1/imunologia , Proteínas do Envelope Viral/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/uso terapêutico , Anticorpos Antivirais/imunologia , Especificidade de Anticorpos , Sítios de Ligação/genética , Células COS , Chlorocebus aethiops , Epitopos/química , Epitopos/genética , Feminino , Herpes Simples/imunologia , Herpes Simples/virologia , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/patogenicidade , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Mutação , Células Vero , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genéticaRESUMO
Human parvovirus 4 (PARV4) is a recently identified virus whose biology, epidemiology and pathogenic potential have yet to be determined. Recently, it was reported that PARV4 DNA persists in tissues of some HIV-infected individuals, whilst PARV4 DNA was not detected in tissues of subjects not infected with HIV. In the present study, liver tissue from 87 individuals, none of who were infected with HIV, with the exception of a single subject, was analyzed for the presence of PARV4 DNA. Overall, PARV4 DNA was detected in 13 specimens (15%). In other tissues examined, PARV4 genotype 2 (also termed PARV5) DNA was detected in one of four paired bone marrow specimens. Tissue viral loads did not exceed 100 copies per microg of genomic DNA. In addition, serum samples from 40 of these individuals all tested negative for PARV4 DNA. In the subjects analyzed in this study, PARV4 genotype 2 appeared to be genetically more heterogeneous than PARV4 genotype 1. The results show that PARV4 DNA can be detected in liver, and that infection with PARV4 is not restricted to HIV-infected individuals. Previous studies showing the presence of PARV4 in plasma, suggest that during infection with PARV4, a viraemic stage occurs, allowing systemic spread to a variety of tissues.