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1.
Kugeler KJ Gurfield N Creek JG Mahoney KS Versage JL Petersen JM 《Applied and environmental microbiology》2005,71(11):7594-7597
The presence of Francisella-like endosymbionts in tick species known to transmit tularemia poses a potential diagnostic problem for laboratories that screen tick samples by PCR for Francisella tularensis. Tick samples initially considered positive for F. tularensis based on standard 16S rRNA gene PCR were found to be positive only for Francisella-like endosymbionts using a multitarget F. tularensis TaqMan assay (ISFtu2, tul4, and iglC) and 16S rRNA gene sequencing. Specificity of PCR-based diagnostics for F. tularensis should be carefully evaluated with appropriate specimen types prior to diagnostic use. 相似文献
2.
Ostland VE Stannard JA Creek JJ Hedrick RP Ferguson HW Carlberg JM Westerman ME 《Diseases of aquatic organisms》2006,72(2):135-145
The present study identifies an emerging disease associated with an aquatic Francisella-like bacterium that can cause mortality in hybrid striped bass Morone chrysops x M. saxatilis reared intensively in freshwater. Clinically affected fish were lethargic, had scattered haemorrhagic cutaneous lesions and diffuse gill pallor. The head kidney and spleen were markedly swollen and contained numerous interstitial granulomas; histological examination revealed small, pleomorphic Gram-negative coccobacilli within vacuolated cells. The bacterium could not be cultured from head kidney homogenates either with standard or enriched microbiological media or following inoculation of a Chinook salmon embryo (CHSE)-214 cell line. No amplification product was obtained from head kidney DNA by polymerase chain reaction (PCR) assay using Piscirickettsia salmonis-specific primers. PCR analysis of infected head kidney homogenate with primers designed for the eubacterial 16S rRNA produced a single amplicon. Phylogenetic analysis of this DNA sequence demonstrated that the sequence aligned most closely with members of the genus Francisella, identified from tilapia Oreochromis spp. in Taiwan and an aquatic Francisella species that was recently isolated from the three-line grunt Parapristipoma trilineatum in Japan. This Francisella-like disease was transmitted to naive hybrid striped bass fingerlings by intraperitoneal injection of tissue homogenates prepared from a natural outbreak. All fish developed gross and histological lesions identical to those from natural outbreaks. Intracellular Gram-negative bacteria were observed within the cytoplasm of cells (presumably macrophages) within the granulomas, but bacteria were not recovered. The 16S DNA sequence of the bacterium obtained from tissues of experimentally infected fish was identical to that obtained from the fish used as infected donor tissue. 相似文献
3.
Maize amylose, separated from granular starch by means of an aqueous leaching process, was used to investigate spherulite formation from concentrated mixtures of starch in water. Amylose (10-20%, w/w) was found to form a spherulitic semicrystalline morphology over a wide range of cooling rates (1-250 degrees C/min), provided it was first heated to >170 degrees C. This is explained through the effect of temperature on chain conformation. A maximum quench temperature of approximately 70 degrees C was required to produce spherulitic morphology. Quench temperatures between 70 and 110 degrees C produced a gel-like morphology. This is explained on the basis of the relative kinetics of liquid-liquid phase separation vis-à-vis crystallization. The possibility of the presence of a liquid crystalline phase affecting the process of spherulite formation is discussed. 相似文献
4.
Vincent IM Creek DJ Burgess K Woods DJ Burchmore RJ Barrett MP 《PLoS neglected tropical diseases》2012,6(5):e1618
A non-targeted metabolomics-based approach is presented that enables the study of pathways in response to drug action with the aim of defining the mode of action of trypanocides. Eflornithine, a polyamine pathway inhibitor, and nifurtimox, whose mode of action involves its metabolic activation, are currently used in combination as first line treatment against stage 2, CNS-involved, human African trypanosomiasis (HAT). Drug action was assessed using an LC-MS based non-targeted metabolomics approach. Eflornithine revealed the expected changes to the polyamine pathway as well as several unexpected changes that point to pathways and metabolites not previously described in bloodstream form trypanosomes, including a lack of arginase activity and N-acetylated ornithine and putrescine. Nifurtimox was shown to be converted to a trinitrile metabolite indicative of metabolic activation, as well as inducing changes in levels of metabolites involved in carbohydrate and nucleotide metabolism. However, eflornithine and nifurtimox failed to synergise anti-trypanosomal activity in vitro, and the metabolomic changes associated with the combination are the sum of those found in each monotherapy with no indication of additional effects. The study reveals how untargeted metabolomics can yield rapid information on drug targets that could be adapted to any pharmacological situation. 相似文献
5.
Discrimination between Francisella tularensis and Francisella-Like Endosymbionts when Screening Ticks by PCR
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Kiersten J. Kugeler Nikos Gurfield Jean G. Creek Kerry S. Mahoney Jessica L. Versage Jeannine M. Petersen 《Applied microbiology》2005,71(11):7594-7597
The presence of Francisella-like endosymbionts in tick species known to transmit tularemia poses a potential diagnostic problem for laboratories that screen tick samples by PCR for Francisella tularensis. Tick samples initially considered positive for F. tularensis based on standard 16S rRNA gene PCR were found to be positive only for Francisella-like endosymbionts using a multitarget F. tularensis TaqMan assay (ISFtu2, tul4, and iglC) and 16S rRNA gene sequencing. Specificity of PCR-based diagnostics for F. tularensis should be carefully evaluated with appropriate specimen types prior to diagnostic use. 相似文献
6.
7.
Macromolecular rate theory (MMRT) provides a thermodynamics rationale to underpin the convergent temperature response in plant leaf respiration
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Liyin L. Liang Vickery L. Arcus Mary A. Heskel Odhran S. O'Sullivan Lasantha K. Weerasinghe Danielle Creek John J. G. Egerton Mark G. Tjoelker Owen K. Atkin Louis A. Schipper 《Global Change Biology》2018,24(4):1538-1547
Temperature is a crucial factor in determining the rates of ecosystem processes, for example, leaf respiration (R) – the flux of plant respired CO2 from leaves to the atmosphere. Generally, R increases exponentially with temperature and formulations such as the Arrhenius equation are widely used in earth system models. However, experimental observations have shown a consequential and consistent departure from an exponential increase in R. What are the principles that underlie these observed patterns? Here, we demonstrate that macromolecular rate theory (MMRT), based on transition state theory (TST) for enzyme‐catalyzed kinetics, provides a thermodynamic explanation for the observed departure and the convergent temperature response of R using a global database. Three meaningful parameters emerge from MMRT analysis: the temperature at which the rate of respiration would theoretically reach a maximum (the optimum temperature, Topt), the temperature at which the respiration rate is most sensitive to changes in temperature (the inflection temperature, Tinf) and the overall curvature of the log(rate) versus temperature plot (the change in heat capacity for the system, ). On average, the highest potential enzyme‐catalyzed rates of respiratory enzymes for R are predicted to occur at 67.0 ± 1.2°C and the maximum temperature sensitivity at 41.4 ± 0.7°C from MMRT. The average curvature (average negative ) was ?1.2 ± 0.1 kJ mol?1 K?1. Interestingly, Topt, Tinf and appear insignificantly different across biomes and plant functional types, suggesting that thermal response of respiratory enzymes in leaves could be conserved. The derived parameters from MMRT can serve as thermal traits for plant leaves that represent the collective temperature response of metabolic respiratory enzymes and could be useful to understand regulations of R under a warmer climate. MMRT extends the classic TST to enzyme‐catalyzed reactions and provides an accurate and mechanistic model for the short‐term temperature response of R around the globe. 相似文献
8.
Rebecca Bullard-Dillard Kim E. Creek Walter A. Scrivens James M. Tour 《Bioorganic chemistry》1996,24(4):376-385
This paper describes thein vivobehavior and potential metabolism of C60and a more water-soluble quaternary ammonium salt-derivatized C60. In both cases, a14C-labeled fullerene core was utilized for the target molecules that were intravenously injected into female Sprague–Dawley rats. The14C-labeled C60(*C60) was rapidly (within 1 min) cleared from the circulation and the majority of the *C60accumulated in the liver (90–95%). *C60was not eliminated from the liver over the 120-h period of this study. Our results also suggest that C60is not metabolized by the typical oxidative patterns characteristic of other polycyclic aromatics. Therefore, although not acutely toxic, use of C60, or its derivatives that could be cleaved back to the parent C60in vivo, would likely lead to long-term fullerene accumulation in the liver. The uptake of *C60and14C-labeled ammonium salt-derivatized C60(1)by human keratinocytesin vitroshowed that while both *C60and1are readily taken up by cells,1accumulates more slowly. Additionally, while C60, at rather high concentrations (2.0 μM) and over extended periods of time (8 days), is able to inhibit the growth of human keratinocytes by about 50%, this effect showed little, if any, photoinducability. 相似文献
9.
Daniel Stoessel Cameron J. Nowell Amy J. Jones Lori Ferrins Katherine M. Ellis Jennifer Riley Raphael Rahmani Kevin D. Read Malcolm J. McConville Vicky M. Avery Jonathan B. Baell Darren J. Creek 《Metabolomics : Official journal of the Metabolomic Society》2016,12(7):126
Introduction
Trypanosoma brucei is the causative agent of human African trypanosomiasis, which is responsible for thousands of deaths every year. Current therapies are limited and there is an urgent need to develop new drugs. The anti-trypanosomal compound, 3-(oxazolo[4,5-b]pyridine-2-yl)anilide (OXPA), was initially identified in a phenotypic screen and subsequently optimized by structure–activity directed medicinal chemistry. It has been shown to be non-toxic and to be active against a number of trypanosomatid parasites. However, nothing is known about its mechanism of action.Objective
Here, we have utilized an untargeted metabolomics approach to investigate the biochemical effects and potential mode of action of this compound in T. brucei.Methods
Total metabolite extracts were analysed by HILIC-chromatography coupled to high resolution mass spectrometry.Results
Significant accumulation of ceramides was observed in OXPA-treated T. brucei. To further understand drug-induced changes in lipid metabolism, a lipidomics method was developed which enables the measurement of hundreds of lipids with high throughput and precision. The application of this LC–MS based approach to cultured bloodstream-form T. brucei putatively identified over 500 lipids in the parasite including glycerophospholipids, sphingolipids and fatty acyls, and confirmed the OXPA-induced accumulation of ceramides. Labelling with BODIPY-ceramide further confirmed the ceramide accumulation following drug treatment.Conclusion
These findings clearly demonstrate perturbation of ceramide metabolism by OXPA and indicate that the sphingolipid pathway is a promising drug target in T. brucei.10.
Donata Rimoldi Kim E. Creek Luigi M. De Luca 《Molecular and cellular biochemistry》1990,93(2):129-140
Summary The molecular mechanism of reduced incorporation of radioactively labeled mannose into hamster liver glycoconjugates during the progression of vitamin A deficiency was investigated. In particular the in vivo incorporation of [2-3H]mannose into GDP-mannose, dolichyl phosphate mannose (Dol-P-Man), lipid-linked oligosaccharides, and glycopeptides of hamster liver was examined. Hamsters maintained on a vitamin A-free diet showed a reduction in the incorporation of mannose into GDP-mannose about 10 days before clinical signs of vitamin A deficiency could be observed. The decrease in [2-3H]mannose incorporated into GDP-mannose was accompanied by a reduction in label incorporated into Dol-P-Man, lipid linked oligosaccharides and glycopeptides, which became more severe with the progression of vitamin A deficiency. By the time they reached a plateau stage of growth, hamsters fed the vitamin A-free diet showed a 50% reduction in the amount of [2-3H]mannose converted to GDP-mannose, and the radioactivity associated with Dol-P-Man and glycopeptides was reduced by approximately 60% as compared to retinoic acid-supplemented controls. These results strongly indicate that the reduced incorporation of mannose into lipidic intermediates and glycoproteins observed during vitamin A deficiency is due to impaired GDP-mannose synthesis.Abbreviations Dol-P-Man
Dolichyl Phosphate Mannose
- Dol-P
Dolichyl Phosphate 相似文献