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951.
A method for estimation of 14CO2 present in blood and tissular samples is described. It is basically based on the introduction of large amounts of a gas mixture (95% O2, 5% CO2) in the samples which serves to remove the CO2 label by gas dilution. The gas phase is later captured in scintillation vials containing an organic-soluble base that retains the carbon label. The results obtained by means of this methodology show much better recoveries for blood samples than those obtained when the classic acid-diffusion method is used. In addition, it is a very fast procedure which does not alter the pH or protein integrity of the biological sample. 相似文献
952.
Diana Marcela Penarete-Vargas Ana?s Boisson Serge Urbach Hervé Chantelauze Suzanne Peyrottes Laurent Fraisse Henri J. Vial 《PloS one》2014,9(12)
Plasmodium falciparum is responsible for severe malaria which is one of the most prevalent and deadly infectious diseases in the world. The antimalarial therapeutic arsenal is hampered by the onset of resistance to all known pharmacological classes of compounds, so new drugs with novel mechanisms of action are critically needed. Albitiazolium is a clinical antimalarial candidate from a series of choline analogs designed to inhibit plasmodial phospholipid metabolism. Here we developed an original chemical proteomic approach to identify parasite proteins targeted by albitiazolium during their native interaction in living parasites. We designed a bifunctional albitiazolium-derived compound (photoactivable and clickable) to covalently crosslink drug–interacting parasite proteins in situ followed by their isolation via click chemistry reactions. Mass spectrometry analysis of drug–interacting proteins and subsequent clustering on gene ontology terms revealed parasite proteins involved in lipid metabolic activities and, interestingly, also in lipid binding, transport, and vesicular transport functions. In accordance with this, the albitiazolium-derivative was localized in the endoplasmic reticulum and trans-Golgi network of P. falciparum. Importantly, during competitive assays with albitiazolium, the binding of choline/ethanolamine phosphotransferase (the enzyme involved in the last step of phosphatidylcholine synthesis) was substantially displaced, thus confirming the efficiency of this strategy for searching albitiazolium targets. 相似文献
953.
954.
Anne Durand Fabien Chauveau Tae-Hee Cho Radu Bolbos Jean-Baptiste Langlois Laure Hermitte Marlène Wiart Yves Berthezène Norbert Nighoghossian 《PloS one》2012,7(11)
Injection of thrombin into the middle cerebral artery (MCA) of mice has been proposed as a new model of thromboembolic stroke. The present study used sequential multiparametric Magnetic Resonance Imaging (MRI), including Magnetic Resonance Angiography (MRA), Diffusion-Weighted Imaging (DWI) and Perfusion-Weighted Imaging (PWI), to document MCA occlusion, PWI-DWI mismatch, and lesion development. In the first experiment, complete MCA occlusion and reproducible hypoperfusion were obtained in 85% of animals during the first hour after stroke onset. In the second experiment, 80% of animals showed partial to complete reperfusion during a three-hour follow-up. Spontaneous reperfusion thus contributed to the variability in ischemic volume in this model. The study confirmed the value of the model for evaluating new thrombolytic treatments, but calls for extended MRI follow-up at the acute stage in therapeutic studies. 相似文献
955.
A L Loubatières G Valette G Ribes A M Rondot 《Comptes rendus des séances de la Société de biologie et de ses filiales》1976,170(4):820-824
In the normal anesthetized dog a continuous perfusion of sodium dichloroacetate (30 mg/kg.h) prevents the increase in blood lactates and pyruvates as well as the lowering of the arterial pH induced by the intraduodenal administration of phenformin (30 mg/kg). Furthermore a perfusion of sodium dichloroacetate (7.5 mg/kg.mn) during 20 minutes, three hours after the administration of phenformin, tends to improve the utilization of blood lactates and pyruvates. 相似文献
956.
Clostridium difficile (C. diff) is one of the most common and most severe hospital-acquired infections; its consequences range from lengthened hospital stay to outright lethality. C. diff causes cellular damage through the action of two large toxins TcdA and TcdB. Recently, there has been increased effort toward developing antitoxin therapies, rather than antibacterial treatments, in hopes of mitigating the acquisition of drug resistance. To date, no analysis of the recognition mechanism of TcdA or TcdB has been attempted. Here, we use small molecule flexible docking followed by unbiased molecular dynamics to obtain a more detailed perspective on how inhibitory peptides, exemplified by two species HQSPWHH and EGWHAHT function. Using principal component analysis and generalized masked Delaunay analysis, an examination of the conformational space of TcdB in its apo form as well as forms bound to the peptides and UDP-Glucose was performed. Although both species inhibit by binding in the active site, they do so in two very different ways. The simulations show that the conformational space occupied by TcdB bound to the two peptides are quite different and provide valuable insight for the future design of toxin inhibitors and other enzymes that interact with their substrates through conformational capture mechanisms and thus work by the disruption of the protein’s intrinsic motions. 相似文献
957.
958.
Daytime variation in T-cell-mediated immunity of Eurasian kestrel Falco tinnunculus nestlings 总被引:1,自引:1,他引:0
Jesús Martínez-Padilla 《Journal of avian biology》2006,37(5):419-424
Host-parasite interactions are central in evolutionary and behavioural ecology. In the last few years, skin injections of the mitogen Phytohaemagglutinin (PHA) have become one of the most important and widely used in-vivo assays of immune function in birds. However, there are no studies of the circadian variation suggesting that care should be taken interpreting results when using this technique. This 3-year study assessed PHA responses as a function of daylight time in 310 Eurasian kestrel Falco tinnunculus nestlings at 24 days of age in Central Spain. I found that T-cell-mediated immunity was positively related to nestling mass and varied among years. Controlling for these variables, I also found that T-cell-mediated immunity decreased with the hour of sampling, and that this pattern was consistent between years. In addition, I found that at the end of the day only, T-cell-mediated immunity decreased with brood size. Parasites seem not to be behind this pattern, but I suggest that the cumulative effect of sibling competition during the day might explain the decrease of cellular immunity with the hour of sampling. Thus, I strongly recommend that future studies of cellular immunity should control for this potential source of variation when nestling self-maintenance is evaluated by the PHA-induced skin-swelling response. 相似文献
959.
960.
A S?s?rman P Chartrand M Lavoie D Tardif R Proschek C Lapointe 《Journal of general microbiology》1979,113(2):297-303
A new type of haem-deficient mutant was isolated in Escherichia coli K12 by neomycin selection. The mutant, designated SASX38, accumulated uroporphyrin, coproporphyrin and protoporphyrin. Since it possessed normal ferrochelatase activity, it was assumed to be deficient in protoporphyrinogen oxidase activity. The gene affected in the mutant was designated hemG. Mapping of the hemG gene by phage P1-mediated transduction showed that it was located very close to the chlB gene (frequency of cotransduction 78.7%), between the metE and rha markers. This location is distinct from the other known hem loci in E. coli K12. 相似文献