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31.
Phenotype microarray profiling of Staphylococcus aureus menD and hemB mutants with the small-colony-variant phenotype 下载免费PDF全文
von Eiff C McNamara P Becker K Bates D Lei XH Ziman M Bochner BR Peters G Proctor RA 《Journal of bacteriology》2006,188(2):687-693
Standard biochemical tests have revealed that hemin and menadione auxotrophic Staphylococcus aureus small-colony variants (SCVs) exhibit multiple phenotypic changes. To provide a more complete analysis of the SCV phenotype, two genetically defined mutants with a stable SCV phenotype were comprehensively tested. These mutants, generated via mutations in menD or hemB that yielded menadione and hemin auxotrophs, were subjected to phenotype microarray (PM) analysis of over 1,500 phenotypes (including utilization of different carbon, nitrogen, phosphate, and sulfur sources; growth stimulation or inhibition by amino acids and other nutrients, osmolytes, and metabolic inhibitors; and susceptibility to antibiotics). Compared to parent strain COL, the hemB mutant was defective in utilization of a variety of carbon sources, including Krebs cycle intermediates and compounds that ultimately generate ATP via electron transport. The phenotype of the menD mutant was similar to that of the hemB mutant, but the defects in carbon metabolism were more pronounced than those seen with the hemB mutant. In both mutant strains, hexose phosphates and other carbohydrates that provide ATP in the absence of electron transport stimulated growth. Other phenotypes of SCV mutants, such as hypersensitivity to sodium selenite, sodium tellurite, and sodium nitrite, were also uncovered by the PM analysis. Key results of the PM analysis were confirmed in independent growth studies and by using Etest strips for susceptibility testing. PM technology is a new and efficient technology for assessing cellular phenotypes in S. aureus. 相似文献
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We compared the histamine responsiveness of canine airways and parenchymal tissues in six anesthetized paralyzed open-chest mongrel dogs, partitioning total lung resistance (RL) into airway resistance (Raw) and tissue viscance (Vti). Pressure was measured during tidal breathing (frequency was 0.3 Hz) at the trachea and in three alveolar regions by use of alveolar capsules. Measurements were taken before and after the delivery of increasing concentrations of aerosolized histamine (0.1-30 mg/ml). We found that Vti accounted for 78 +/- 8% of RL under base-line conditions; this proportion remained relatively constant throughout the histamine concentration-response curve. There was a significant correlation between percent change in Vti and percent change in Raw at all levels of histamine-induced constriction (P less than 0.001). Moreover, the sensitivity of the tissues and airways (defined as the concentration of histamine required to double resistance) was remarkably similar. We conclude that, at this frequency of ventilation, Vti accounts for the major portion of RL both under base-line conditions and after histamine-induced constriction. Although increases in RL cannot be attributed solely to events occurring in the airways, the close correlation between changes in Raw and Vti and the similar sensitivities of the two support the use of indexes reflecting changes in airway caliber as an indicator of overall lung histamine responsiveness. 相似文献
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Setzer WN Vogler B Bates RB Schmidt JM Dicus CW Nakkiew P Haber WA 《Phytochemical analysis : PCA》2003,14(1):54-59
A combination of HPLC-MS and HPLC-NMR techniques has been used to analyse the cytotoxic fractions of the dichloromethane extract of bark of Stauranthus perforatus. Six furanocoumarins (byakangelicol, heraclenin, heraclenol, imperatorin, isopimpinellin and xanthotoxin) and nine quinoline alkaloids (two known compounds, veprisine and 5-hydroxy-1-methyl-2-phenyl-4-quinolone, along with seven novel compounds, stauranthine, 3',4'-dihydroxy-3',4'-dihydroveprisine, 3',4'-dihydroxy-3',4'-dihydrostauranthine, 3',6'-dihydroxy-3',6'-dihydroveprisine, 3',6'-dihydroxy-3',6'-dihydrostauranthine, 6'-hydroxy-3'-ketoveprisine and 6'-hydroxy-3'-ketostauranthine) have been identified in the fractions. 相似文献
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Athanasiou A Clarke AB Turner AE Kumaran NM Vakilpour S Smith PA Bagiokou D Bradshaw TD Westwell AD Fang L Lobo DN Constantinescu CS Calabrese V Loesch A Alexander SP Clothier RH Kendall DA Bates TE 《Biochemical and biophysical research communications》2007,364(1):131-137
Time-lapse microscopy of human lung cancer (H460) cells showed that the endogenous cannabinoid anandamide (AEA), the phyto-cannabinoid Δ-9-tetrahydrocannabinol (THC) and a synthetic cannabinoid HU 210 all caused morphological changes characteristic of apoptosis. Janus green assays of H460 cell viability showed that AEA and THC caused significant increases in OD 595 nm at lower concentrations (10-50 μM) and significant decreases at 100 μM, whilst HU 210 caused significant decreases at all concentrations. In rat heart mitochondria, all three ligands caused significant decreases in oxygen consumption and mitochondrial membrane potential. THC and HU 210 caused significant increases in mitochondrial hydrogen peroxide production, whereas AEA was without significant effect. All three ligands induced biphasic changes in either mitochondrial complex I activity and/or mitochondrial complex II-III activity. These data demonstrate that AEA, THC, and HU 210 are all able to cause changes in integrated mitochondrial function, directly, in the absence of cannabinoid receptors. 相似文献
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Lipemic serum from three patients with acute pancreatitis and type IV hyperlipemia was fractionated into very-low-density lipoproteins and clear serum. Amylase activity (determined by the Phadebas method) in the component fractions did not exceed that in the original lipemic serum. Addition of these fractions or VLDL and chylomicrons from asymptomatic patients with hyperlipemia to nonlipemic serum from patients with "routine acute pancreatitis" did not inhibit amylase activity or alter the electrophoretic mobility of amylase isoenzymes. Therefore the normal amylase activity often observed in hyperlipemic pancreatitis does not result from an inhibition of amylase activity by serum lipoproteins. 相似文献
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Gourfinkel-An I Parain K Hartmann A Mangiarini L Brice A Bates G Hirsch EC 《Journal of neurochemistry》2003,86(6):1369-1378
Huntington's disease is an autosomal dominant disorder with degeneration of medium size striatal neurones. As the disease evolves, other neuronal populations are also progressively affected. A transgenic mouse model of the disease (R6/2) that expresses exon 1 of the human Huntington gene with approximately 150 CAG repeats has been developed, but GABA concentrations are reported to be normal in the striatum of these animals. In the present study, we analysed the status of GABAergic systems by means of glutamic acid decarboxylase (GAD)67 mRNA in situ hybridization in the brain of R6/2 transgenic mice and wild-type littermates. We show that GAD67 expression is normal in the striatum, cerebellum and septum but decreased in the frontal cortex, parietal cortex, globus pallidus, entopeduncular nucleus and substantia nigra pars reticulata of R6/2 mice. These data, which may, in part, account for the behavioural changes seen in these animals, indicate that at 12.5 weeks of age the pathological features seen in the mice differ from those seen in humans with Huntington's disease. 相似文献
40.
Effects of simulated long-term climatic change on the bryophytes of a limestone grassland community 总被引:2,自引:0,他引:2
The bryophyte vegetation of upland limestone grassland at Buxton in the southern Pennine Hills (UK) was studied following seven years' continuous simulated climate change treatments. The experimental design involved two temperature regimes (ambient, winter warming by 3°C) in factorial combination with three moisture regimes (normal, summer drought, supplemented summer rainfall) and with five replicate blocks. Percentage cover of the bryophytes was estimated visually using 15 randomly positioned quadrats (30 cm × 30 cm) within each of the 30 3 m × 3 m plots. Significant treatment effects were found but these were relatively modest. Total bryophyte cover and cover of Calliergonella cuspidata and Rhytidiadelphus squarrosus responded negatively to drought, whereas Fissidens dubius increased in the droughted plots. Campyliadelphus chrysophyllus increased with winter warming, while R. squarrosus, Lophocolea bidentata and species richness all decreased. The effects on the total bryophyte flora were further studied by canonical correspondence analysis, which yielded a first axis reflecting the combined effects of the moisture and temperature treatments. However, this analysis and a detrended correspondence analysis of the plot data also revealed that natural factors were more important causes of variation in the grassland community than the simulated climate treatments. It was concluded that dewfall may be an important source of moisture for grassland bryophytes and that this factor may have reduced the impact of the moisture treatments. The absence of some thermophilous species such as Homalothecium lutescens in the plots initially may also have reduced their scope for major vegetational change. 相似文献