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Cottyn B Acher F Ramassamy B Alvey L Lepoivre M Frapart Y Stuehr D Mansuy D Boucher JL Vichard D 《Bioorganic & medicinal chemistry》2008,16(11):5962-5973
A series of new 7-monosubstituted and 3,7-disubstituted indazoles have been prepared and evaluated as inhibitors of nitric oxide synthases (NOS). 1H-indazole-7-carbonitrile (6) was found equipotent to 7-nitro-1H-indazole (1) and demonstrated preference for constitutive NOS over inducible NOS. By contrast, 1H-indazole-7-carboxamide (8) was slightly less potent but demonstrated a surprising selectivity for the neuronal NOS. Further substitution of 6 by a Br-atom at carbon-3 of the heterocycle enhanced 10-fold the inhibitory effects. Inhibition of NO formation by 6 appeared to be competitive versus both substrate and the cofactor (6R)-5,6,7,8-tetrahydro-l-biopterin (H(4)B). In close analogies with 1, compound 6 strongly inhibited the NADPH oxidase activity of nNOS and induced a spin state transition of the heme-Fe(III). Our results are explained with the help of the X-ray structures that identified key-features for binding of 1 at the active site of NOS. 相似文献
184.
Tropical hardwood hammocks, among the rarest and most threatened vegetative communities in the United States, occur throughout the 225-km Florida Keys archipelago as it extends toward the Caribbean from the southeast tip of peninsular Florida. Compounding their critical conservation status, tropical hardwood hammocks and the dynamics that support their peculiar species diversity in the region are poorly understood. The goal of this study was to explore the dynamics of the species compositional gradient of the hammocks along the Florida Keys, and to identify significant ecological units within the gradient. The primary data for this research were assembled from the Institute for Regional Conservation's floristic database of South Florida. We were able to extract presence/absence data for 295 species from comprehensive surveys of 23 study sites. Nonmetric multidimensional scaling was used to deconstruct the compositional trends into a reduced ordination space. Cluster analysis was subsequently used to identify discrete ecological units. Additionally, we used vector fitting to interpret the significant correlated ancillary variables. Our main results were three well-fitted nonmetric multidimensional scaling axes with three nonoverlapping ecological units. Of the ancillary variables, latitude, longitude, percent composition from biogeographical regions, richness, and area were correlated to the nonmetric multidimensional scaling results. These results increase our understanding of the community structure of the hammocks along the Florida Keys, and can contribute to increasing our ability to adequately protect and restore tropical hardwood hammocks and other similar tropical dry forest communities throughout the Caribbean. 相似文献
185.
Ethidium monoazide bromide (EMA) treatment of pure culture and environmental waters at low concentrations (1.0–7.5 μg/ml)
indicated effective enumeration of viable and viable but nonculturable Escherichia coli in pure cultures, creek waters, and secondary activated sludge effluent samples by quantitative polymerase chain reaction
(qPCR) amplification of the uidA and fliC gene targets at turbidity values <10 NTU. However, EMA treatment was not effective in primary clarifier and secondary trickling
filter effluents where turbidities were ≥10 NTU. In viable pure cultures, rapidly dividing and senescent cells were most affected
by increasing EMA concentrations. Amplification of heat-killed pure bacterial cultures decreased 4 to 6 logs depending on
EMA concentration and culture age. The greatest difference was observed in 5-h cultures using 7.5 μg/ml EMA. Turbidity (≥100
NTU) in environmental samples inhibited EMA effectiveness on viability discrimination. Enumeration of E. coli in certain wastewaters using EMA-qPCR was similar to culture suggesting that EMA treatment could be incorporated into qPCR
assays for the quantification of viable bacteria increasing assay time no more than 30 min. Our results indicate that EMA
can be used in routine qPCR assays, but optimum conditions for exposure must be identified for each sample type due to sample
matrix effects such as turbidity. 相似文献
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Bath JL Robinson M Kennedy MW Agbasi C Linz L Maetzold E Scheidt M Knox M Ram D Hein J Clark C Drees J 《Journal of nematology》2009,41(3):228-233
Hp-FAR-1 is a major, secreted antigen of the parasitic nematode Heligmosomoides polygyrus, a laboratory mouse model frequently used to study the cellular mechanisms of chronic helminth infections. The DNA encoding Hp-FAR-1 was recovered by screening a fourth larval (L(4)) H. polygyrus cDNA expression library using antibodies raised against L(4) stage excretory/secretory (E/S) proteins. Predictions of secondary structure based on the Hp-FAR-1 amino acid sequence indicated that an alpha-helix predominates in Hp-FAR-1, possibly with some coiled-coil conformation, with no beta-structure. Fluorescence-based ligand binding analysis confirmed that the recombinant Hp-FAR-1 (rHp-FAR-1) binds the fluorescent fatty acid analog 11-((5-[dimethylaminoaphthalene-1-sulfonyl)amino)undecanoic acid (DAUDA), and by competition oleic acid. RT-PCR amplification of the hp-far-1 gene indicated that the gene is transcribed in all parasitic stages of the organism's life cycle. The presence of a secreted FAR protein in the well-defined laboratory model of H. polygyrus provides an excellent model for the further study and analysis of the in vivo role of secreted FAR proteins in parasitism, and supports the mounting evidence that secreted FAR proteins play a major role in nematode parasitism. 相似文献
189.
Frederic Dalle Betty Wchtler Coralie L'Ollivier Gudrun Holland Norbert Bannert Duncan Wilson Catherine Labrure Alain Bonnin Bernhard Hube 《Cellular microbiology》2010,12(2):248-271
The human pathogenic fungus Candida albicans can cause systemic infections by invading epithelial barriers to gain access to the bloodstream. One of the main reservoirs of C. albicans is the gastrointestinal tract and systemic infections predominantly originate from this niche. In this study, we used scanning electron and fluorescence microscopy, adhesion, invasion and damage assays, fungal mutants and a set of fungal and host cell inhibitors to investigate the interactions of C. albicans with oral epithelial cells and enterocytes. Our data demonstrate that adhesion, invasion and damage by C. albicans depend not only on fungal morphology and activity, but also on the epithelial cell type and the differentiation stage of the epithelial cells, indicating that epithelial cells differ in their susceptibility to the fungus. C. albicans can invade epithelial cells by induced endocytosis and/or active penetration. However, depending on the host cell faced by the fungus, these routes are exploited to a different extent. While invasion into oral cells occurs via both routes, invasion into intestinal cells occurs only via active penetration. 相似文献
190.
Liu BP Cafferty WB Budel SO Strittmatter SM 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2006,361(1473):1593-1610
Robust axonal growth is required during development to establish neuronal connectivity. However, stable fibre patterns are necessary to maintain adult mammalian central nervous system (CNS) function. After adult CNS injury, factors that maintain axonal stability limit the recovery of function. Extracellular molecules play an important role in preserving the stability of the adult CNS axons and in restricting recovery from pathological damage. Adult axonal growth inhibitors include a group of proteins on the oligodendrocyte, Nogo-A, myelin-associated glycoprotein, oligodendrocyte-myelin glycoprotein and ephrin-B3, which interact with axonal receptors, such as NgR1 and EphA4. Extracellular proteoglycans containing chondroitin sulphates also inhibit axonal sprouting in the adult CNS, particularly at the sites of astroglial scar formation. Therapeutic perturbations of these extracellular axonal growth inhibitors and their receptors or signalling mechanisms provide a degree of axonal sprouting and regeneration in the adult CNS. After CNS injury, such interventions support a partial return of neurological function. 相似文献