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1.
In prokaryote organisms, N-glycosylation of proteins is often correlated to cell–cell recognition and extracellular events. Those glycoproteins are potential targets for infection control. To date, many surface-glycosylated proteins from bacterial pathogens have been described. However, N-linked Pseudomonas surface-associated glycoproteins remain underexplored. We report a combined enrichment and labeling strategy to identify major glycoproteins on the outside of microorganisms. More precisely, bacteria were exposed to a mix of biotinylated lectins able to bind with glycoproteins. The latter were then recovered by avidin beads, digested with trypsin, and submitted to mass spectrometry. The targeted mixture of glycoproteins was additionally deglycosylated in the presence of H2 18O to incorporate 18O during PNGase F treatment and were also analyzed using mass spectrometry. This approach allowed us to identify a few tens of potential N-glycoproteins, among which flagellin FliC was the most abundant. To detect the possible sites of FliC modifications, a de novo sequencing step was also performed to discriminate between spontaneous deamidation and N-glycan loss. This approach led to the proposal of three potential N-glycosylated sites on the primary sequence of FliC: N26, N69, and N439, with two of these three asparagines belonging to an N-X-(S/T) consensus sequence. These observations suggest that flagellin FliC is a heterogeneous protein mixture containing both O- and N-glycoforms.
Figure
Analytical scenario developed for bacterial glycoprotein enrichment. This strategy includes three main steps: (1) exposure of Pseudomonas aeruginosa cells to a mixture of biotinylated lectins [wheat germ agglutinin (WGA) and concanavalin A (ConA)]; (2) enrichment of N-glycoproteins by elution with avidin beads; and (3) mass spectrometry (MS) identification and characterization of intact and deglycosylated peptides before and after H2 18O PNGase F enzymatic treatment, respectively  相似文献   
2.
We show that the equation Δu = p(x)f(u) has a positive solution on R N , N ≥ 3, satisfying <artwork name="GAPA31011ei1"> <artwork name="GAPA31011ei2"> if and only if <artwork name="GAPA31011ei3"> when ψ(r) = min{p(x): |x| = r}. The nondecreasing continuous function f satisfies f(0) = 0, f (s) > 0 for s > 0, and sup s ≥ 1 f(s)/s<∞, and the nonnegative continuous function p is required to be asymptotically radial. This extends previous results which required the function p to be constant or radial.  相似文献   
3.
With the increased availability of information and communications technology (ICT) in schools, it is important to examine how teachers use it in their instruction. This paper describes a study that involved a number of secondary mathematics teachers who had been using the new technology in their teaching for a number of years. It reports on some of the factors affecting teacher’ use of the new technology identified by analysing the data collected through questionnaires and teachers’ interviews. A possible theoretical framework to account for teachers’ learning about ICT and teachers’ incorporation of ICT into their planning for teaching mathematics is presented.  相似文献   
4.
In the present study, we evaluated the antioxidant and the scavenging ability of C14, C15 and C16 bacillomycin D-like lipopeptides produced by B38 strain. They all displayed strong reducing power activity, hydroxyl and superoxide anion radicals scavenging activities and inhibition of lipid peroxidation. In addition, they were found to protect plasmid DNA damage from hydroxyl radical oxidation. Data suggested that their antioxidant potency can be attributed to the hydrophobic and aromatic side-chain groups of their amino acids as well as to the aliphatic chain of their beta amino fatty acids. Note that the hydrocarbon chain length did not interfere with the antioxidant power. Overall, such bacillomycin D lipopeptides which exhibit antioxidant and radical scavenging activities may be useful for cosmetic, therapeutic or pharmaceutical purposes in order to delay or prevent oxidative deterioration of manufactured products.  相似文献   
5.
Small fiber peripheral neuropathy is an early complication of diabetes. Electric skin response to some stimulus, as electrochemical skin conductance ECS, is a promising route in the early follow‐up of such diseases. It is related to sweat gland innervations and their permeability to chlorides and protons; it is non‐invasive, quantitative and reproducible. In routine clinical use, it could allow to better adapt the treatments and improve the adhesion for preventing pathological progress, thus reducing colossal healthcare costs. To optimize the measurements and understand the electrochemical behavior of electrodes, an original electrolytic cell was designed in lab scale. Thereby, an electrolyte is chosen to mimic sweat composition. For achieving currents range of ESC in vivo measurements, the original idea was to play on electrolyte viscosity by adding sucrose. In this paper, the novel electrolytic lab cell is presented with its limiting kinetics processes. A model of chloride migration to the anode and global electric model dedicated to the cell are proposed. Cell parameters are thoroughly studied, e. g. the resistance, which is equivalent to the inverse of ESC, by exploiting the models and through in vitro experiments, with protocols focusing on reproducibility. This original approach establishes, inter alia, an important result: the resistance is accurately retrieved using linear voltammetry, whereas single voltage measurement fails notably and is, therefore, unsatisfactory.  相似文献   
6.
The alternate deposition of polyanions and polycations leads to the formation of films called polyelectrolyte multilayer films (PEMs). Two types of growth processes are reported in the literature, leading to films that grow either linearly or exponentially with the number of deposition steps. In this article we try to establish a correlation between the nature of the growth process and the heat of complexation between the polyanions and the polycations constituting the PEM film. Isothermal titration microcalorimetry experiments performed on several polyanion/polycation systems seem to indicate that an endothermic complexation process is characteristic of an exponential film growth, whereas a strongly exothermic process corresponds to a linear growth regime. Finally, weakly exothermic processes seem to be associated with weakly exponentially growing films. These results thus show that exponentially growing processes are mainly driven by entropy. This explains why the exponential growth processes are more sensitive to temperature than the linear growing processes. This temperature sensitivity is shown on the poly-L-glutamic acid/poly(allylamine) system which grows either linearly or exponentially depending on the ionic strength of the polyelectrolyte solutions.  相似文献   
7.
Electrochemical synthesis of ceria films was performed on a stainless steel substrate in view of Solid Oxide Fuel Cells (SOFC) applications. Films were obtained from aqueous nitrate solutions via cathodic deposition method at room temperature. A constant potential value of −0.8 V/(SCE) was applied to reduce the molecular oxygen as hydroxide precursor, leading to a formation of adherent, homogeneous and covering films in 20 min deposition time. Structure, morphology and composition of as-grown coatings were studied by X-ray diffraction, Raman and energy-dispersive X-ray spectroscopy, as well as scanning electron microscopy. Cubic fluorite-type nanostructured ceria of leaf-like particles was synthesized. Thermal annealing (600°C, 1 h) was found to enhance ceria crystallinity.  相似文献   
8.
9.
When cultured in minimal growth medium, the B38 strain of Bacillus subtilis did not exhibit any antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) clinical isolate. Coculturing B38 strain with viable MRSA cells weakly increased antibacterial activity production (20 AU/ml). Addition of dead MRSA cells in a B38 culture, increased by 8-fold the B. subtilis strain antibacterial activity reaching 160 AU/ml against MRSA strain. This antibacterial activity recovered from cell-free supernatants was stimulated by an autoinducing compound which is sensitive to the action of proteinase K suggesting a proteinaceous nature. This compound was heat-stable till 80 °C and showed a molecular mass around 20 kDa as determined by SDS-PAGE. These results suggest that the production of antibacterial compounds by B38 strain is dependent on the amount of the autoinducing compound.  相似文献   
10.
Two strains (an environmental strain and a reference one coming from a national culture collection) of Yersinia ruckeri, a fish pathogenic bacterium, are characterised according to the ability to adhere on wood, concrete, polyvinylchloride (PVC) and fibreglass, four materials commonly found in fish farms. The relationships between adherence, bacterial and support hydrophobicities and surface roughness are investigated. The results show that: (i) Y. ruckeri is strongly hydrophilic; (ii) the environmental strain exhibits a higher ability to adhere than the reference one; and (iii) for the two strains a strong correlation is observed between roughness amplitude (RA) of the support material and adhesion ability.  相似文献   
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