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A heterotrophic nitrifying Alcaligenes sp. from soil was grown as a denitrifier on nitrate and subjected to oxidant pulse experiments to ascertain the apparent effeciencies of proton translocations during O2 and nitrogen-oxide respirations. With endogenous substrate as the reducing agent the H+/2e– ratios, extrapolated to zero amount of oxidant per pulse, were 9.4, 3.7, 4.3 and 3.5 for O2, nitrate, nitrite and N2O, respectively. The value for O2 and those for the N-oxides are, respectively, somewhat larger and smaller than corresponding values for Paracoccus denitrificans. None of the three permeant ions employed with the Alcaligenes sp. (valinomycin-K+, thiocyanate and triphenylmethylphosphonium) was ideal for all purposes. Thiocyanate provided highest ratios for O2 but abolished the oxidant pulse response for nitrate and N2O. Valinomycin was slow to penetrate to the cytoplasmic membrane and relatively high concentrations were required for optimal performance. Triphenylmethylphosphonium enhanced passive proton permeability and diminished proton translocation at concentrations required to realize the maximal oxidant pulse response. 相似文献
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Don Farthing Lynne Gehr H. Thomas Karnes Domenic Sica Todd Gehr Terri Larus 《Biomarkers》2013,18(6):623-634
AbstractAcetyl salicylic acid (aspirin) is one of the most widely used drugs in the world. Various plasma concentrations of aspirin and its predominant metabolite, salicylic acid, are required for its antiarthritic (1.5–2.5 mM), anti-inflammatory (0.5–5.0 mM) or antiplatelet (0.18–0.36 mM) actions. A recent study demonstrated the inhibitory effects of both aspirin and salicylic acid on oxidative phosphorylation and ATP synthesis in isolated rat cardiac mitochondria in a dose-dependent manner (0–10 mM concentration range). In this context, the present study was conducted to determine the effects of salicylic acid on inosine efflux (a potential biomarker of acute cardiac ischaemia) as well as cardiac contractile function in the isolated mouse heart following 20 min of zero-flow global ischaemia. Inosine efflux was found at significantly higher concentrations in ischaemic hearts perfused with Krebs buffer fortified with 1.0 mM salicylic acid compared with those without salicylic acid (12575±3319 vs. 1437±348 ng ml?1 min?1, mean±SEM, n=6 per group, p<0.01). These results indicate that 1.0 mM salicylic acid potentiates 8.8-fold ATP nucleotide purine catabolism into its metabolites (e.g. inosine, hypoxanthine). Salicylic acid (0.1 or 1.0 mM) did not appreciably inhibit purine nucleoside phosphorylase (the enzyme converts inosine to hypoxanthine) suggesting the augmented inosine efflux was due to the salicylic acid effect on upstream elements of cellular respiration. Whereas post-ischaemic cardiac function was further depressed by 1.0 mM salicylic acid, perfusion with 0.1 mM salicylic acid led to a remarkable functional improvement despite moderately increased inosine efflux (2.7-fold). We conclude that inosine is a sensitive biomarker for detecting cardiac ischaemia and salicylic acid-induced effects on cellular respiration. However, the inosine efflux level appears to be a poor predictor of the individual post-ischaemic cardiac functional recovery in this ex vivo model. 相似文献
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Christine A. Farthing Don E. Farthing Saisudha Koka Terri Larus Itaf Fakhry Lei Xi Rakesh C. Kukreja Domenic Sica Todd W.B. Gehr 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(28):2891-2895
A simple and sensitive high-performance liquid chromatographic (HPLC) method utilizing fluorescence detection was developed for the determination of the phosphodiesterase type 5 inhibitor tadalafil in mouse plasma. This method utilizes a simple sample preparation (protein precipitation) with high recovery of tadalafil (∼98%), which eliminates the need for an internal standard. For constituent separation, the method utilized a monolithic C18 column and a flow rate of 1.0 mL/min with a mobile phase gradient consisting of aqueous trifluoroacetic acid (0.1% TFA in deionized water pH 2.2, v/v) and acetonitrile. The method calibration was linear for tadalafil in mouse plasma from 100 to 2000 ng/mL (r > 0.999) with a detection limit of approximately 40 ng/mL. Component fluorescence detection was achieved using an excitation wavelength of 275 nm with monitoring of the emission wavelength at 335 nm. The intra-day and inter-day precision (relative standard deviation, RSD) values for tadalafil in mouse plasma were less than 14%, and the accuracy (percent error) was within −14% of the nominal concentration. The method was utilized on mouse plasma samples from research evaluating the potential cardioprotective effects of tadalafil on mouse heart tissue exposed to doxorubicin, a chemotherapeutic drug with reported cardiotoxic effects. 相似文献
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This study examines the abundance of the major protein constituents of the pre-replication complex (pre-RC), both genome-wide and in association with specific replication origins, namely the lamin B2, c-myc, 20mer1, and 20mer2 origins. Several pre-RC protein components, namely ORC1-6, Cdc6, Cdt1, MCM4, MCM7, as well as additional replication proteins, such as Ku70/86, 14-3-3, Cdc45, and PCNA, were comparatively and quantitatively analyzed in both transformed and normal cells. The results show that these proteins are overexpressed and more abundantly bound to chromatin in the transformed compared to normal cells. Interestingly, the 20mer1, 20mer2, and c-myc origins exhibited a two- to threefold greater origin activity and a two- to threefold greater in vivo association of the pre-RC proteins with these origins in the transformed cells, whereas the origin associated with the housekeeping lamin B2 gene exhibited both similar levels of activity and in vivo association of these pre-RC proteins in both cell types. Overall, the results indicate that cellular transformation is associated with an overexpression and increased chromatin association of the pre-RC proteins. This study is significant, because it represents the most systematic comprehensive analysis done to date, using multiple replication proteins and different replication origins in both normal and transformed cell lines. 相似文献
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The relationships among suprathreshold taste responses to acesulfame-K, Na-saccharin and 6-n-propylthiouracil (PROP) were examined in two studies. In the first study, the labeled magnitude scale was used with the high anchor labeled as 'strongest imaginable oral sensation' and in the second study, it was labeled as 'strongest imaginable sensation of any kind'. Results from the two procedures were similar. Individual differences among 65 subjects were seen in bitter responses to acesulfame-K and saccharin. Bitter responses to acesulfame-K ands accharin were positively correlated, but showed no significant relationship with responses to PROP bitterness or with PROP taster groups. Saccharin and acesulfame-K may share a common mechanism for bitter taste reception and transduction, one that varies across individuals and is different from mechanisms mediating bitter responses to PROP. Changing the instructions of the labeled magnitude scale induced a context effect. Ratings of sweetness referenced to the 'strongest imaginable sensationof any kind' were lower than ratings referenced to just oral sensations. 相似文献
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Degradation Pathway and Generation of Monohydroxamic Acids from the Trihydroxamate Siderophore Deferrioxamine B
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Agnes Pierwola Tomasz Krupinski Peter Zalupski Michael Chiarelli Domenic Castignetti 《Applied microbiology》2004,70(2):831-836
Siderophores are avid ferric ion-chelating molecules that sequester the metal for microbes. Microbes elicit siderophores in numerous and different environments, but the means by which these molecules reenter the carbon and nitrogen cycles is poorly understood. The metabolism of the trihydroxamic acid siderophore deferrioxamine B by a Mesorhizobium loti isolated from soil was investigated. Specifically, the pathway by which the compound is cleaved into its constituent monohydroxamates was examined. High-performance liquid chromatography and mass-spectroscopy analyses demonstrated that M. loti enzyme preparations degraded deferrioxamine B, yielding a mass-to-charge (m/z) 361 dihydroxamic acid intermediate and an m/z 219 monohydroxamate. The dihydroxamic acid was further degraded to yield a second molecule of the m/z 219 monohydroxamate as well as an m/z 161 monohydroxamate. These studies indicate that the dissimilation of deferrioxamine B by M. loti proceeds by a specific, achiral degradation and likely represents the reversal by which hydroxamate siderophores are thought to be synthesized. 相似文献
8.
NGF controls APP cleavage by downregulating APP phosphorylation at Thr668: relevance for Alzheimer's disease
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Viviana Triaca Valentina Sposato Giulia Bolasco Maria Teresa Ciotti Piergiuseppe Pelicci Amalia C. Bruni Chiara Cupidi Raffaele Maletta Marco Feligioni Robert Nisticò Nadia Canu Pietro Calissano 《Aging cell》2016,15(4):661-672
NGF has been implicated in forebrain neuroprotection from amyloidogenesis and Alzheimer's disease (AD). However, the underlying molecular mechanisms are still poorly understood. Here, we investigated the role of NGF signalling in the metabolism of amyloid precursor protein (APP) in forebrain neurons using primary cultures of septal neurons and acute septo‐hippocampal brain slices. In this study, we show that NGF controls the basal level of APP phosphorylation at Thr668 (T668) by downregulating the activity of the Ser/Thr kinase JNK(p54) through the Tyr kinase signalling adaptor SH2‐containing sequence C (ShcC). We also found that the specific NGF receptor, Tyr kinase A (TrkA), which is known to bind to APP, fails to interact with the fraction of APP molecules phosphorylated at T668 (APPpT668). Accordingly, the amount of TrkA bound to APP is significantly reduced in the hippocampus of ShcC KO mice and of patients with AD in which elevated APPpT668 levels are detected. NGF promotes TrkA binding to APP and APP trafficking to the Golgi, where APP–BACE interaction is hindered, finally resulting in reduced generation of sAPPβ, CTFβ and amyloid‐beta (1‐42). These results demonstrate that NGF signalling directly controls basal APP phosphorylation, subcellular localization and BACE cleavage, and pave the way for novel approaches specifically targeting ShcC signalling and/or the APP–TrkA interaction in AD therapy. 相似文献
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An examination of proton translocation and energy conservation during heterotrophic nitrification 总被引:1,自引:0,他引:1
Whether selected heterotrophic nitrifiers, as do the autotrophs, conserve energy during the oxidation of their nitrogenous substrates was studied. The examination of proton translocation of four different bacterial nitrifiers capable of pyruvic oxime [(PO), CH3-C(NOH)-COOH] nitrification and by an NH4+ oxidizing Arthrobacter sp. was initiated. Three of the PO nitrifying bacteria, all pseudomonads, oxidize hydroxylamine (NH2OH) at a greater rate than PO and yielded only stoichiometric protons when NH2OH was the reductant. The fourth bacterium, Alcaligenes faecalis ATCC 8750, an adept PO oxidizer, does not appreciably oxidize NH2OH. The bacterium displayed----H+NH2OH ratios far less than if NH2OH was stoichiometrically converted to nitrite. When given NH4+, the Arthrobacter sp. yielded proton translocation patterns which were inconsistent with the metabolic data collected concerning NH4+ oxidation. Thus no data was collected which supported energy conservation via proton translocation by these heterotrophic nitrifiers. 相似文献
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