45 research outputs found
Clogging by sieving in microchannels: Application to the detection of contaminants in colloidal suspensions
We report on a microfluidic method that allows measurement of a small
concentration of large contaminants in suspensions of solid micrometer-scale
particles. To perform the measurement, we flow the colloidal suspension through
a series of constrictions, i.e. a microchannel of varying cross-section. We
show and quantify the role of large contaminants in the formation of clogs at a
constriction and the growth of the resulting filter cake. By measuring the time
interval between two clogging events in an array of parallel microchannels, we
are able to estimate the concentration of contaminants whose size is selected
by the geometry of the microfluidic device. This technique for characterizing
colloidal suspensions offers a versatile and rapid tool to explore the role of
contaminants on the properties of the suspensions
Mechanical tuning of the evaporation rate of liquid on crossed fibers
We investigate experimentally the drying of a small volume of perfectly
wetting liquid on two crossed fibers. We characterize the drying dynamics for
the three liquid morphologies that are encountered in this geometry: drop,
column and a mixed morphology, in which a drop and a column coexist. For each
morphology, we rationalize our findings with theoretical models that capture
the drying kinetics. We find that the evaporation rate depends significantly on
the liquid morphology and that the drying of liquid column is faster than the
evaporation of the drop and the mixed morphology for a given liquid volume.
Finally, we illustrate that shearing a network of fibers reduces the angle
between them, changes the morphology towards the column state, and so enhances
the drying rate of a volatile liquid deposited on it
Transcriptome and Proteome Exploration to Model Translation Efficiency and Protein Stability in Lactococcus lactis
This genome-scale study analysed the various parameters influencing protein levels in cells. To achieve this goal, the model bacterium Lactococcus lactis was grown at steady state in continuous cultures at different growth rates, and proteomic and transcriptomic data were thoroughly compared. Ratios of mRNA to protein were highly variable among proteins but also, for a given gene, between the different growth conditions. The modeling of cellular processes combined with a data fitting modeling approach allowed both translation efficiencies and degradation rates to be estimated for each protein in each growth condition. Estimated translational efficiencies and degradation rates strongly differed between proteins and were tested for their biological significance through statistical correlations with relevant parameters such as codon or amino acid bias. These efficiencies and degradation rates were not constant in all growth conditions and were inversely proportional to the growth rate, indicating a more efficient translation at low growth rate but an antagonistic higher rate of protein degradation. Estimated protein median half-lives ranged from 23 to 224 min, underlying the importance of protein degradation notably at low growth rates. The regulation of intracellular protein level was analysed through regulatory coefficient calculations, revealing a complex control depending on protein and growth conditions. The modeling approach enabled translational efficiencies and protein degradation rates to be estimated, two biological parameters extremely difficult to determine experimentally and generally lacking in bacteria. This method is generic and can now be extended to other environments and/or other micro-organisms
The Response of Lactococcus lactis to Membrane Protein Production
Background: The biogenesis of membrane proteins is more complex than that of water-soluble proteins, and recombinant expression of membrane proteins in functional form and in amounts high enough for structural and functional studies is often problematic. To better engineer cells towards efficient protein production, we set out to understand and compare the cellular consequences of the overproduction of both classes of proteins in Lactococcus lactis, employing a combined proteomics and transcriptomics approach.
Methodology and Findings: Highly overproduced and poorly expressed membrane proteins both resulted in severe growth defects, whereas amplified levels of a soluble substrate receptor had no effect. In addition, membrane protein overproduction evoked a general stress response (upregulation of various chaperones and proteases), which is probably due to accumulation of misfolded protein. Notably, upon the expression of membrane proteins a cell envelope stress response, controlled by the two-component regulatory CesSR system, was observed.
Conclusions: The physiological response of L. lactis to the overproduction of several membrane proteins was determined and compared to that of a soluble protein, thus offering better understanding of the bottlenecks related to membrane protein production and valuable knowledge for subsequent strain engineering.
Bimodal dynamics of primary metabolism-related responses in tolerant potato-Potato virus Y interaction
BACKGROUND: Potato virus Y (PVY) is a major pathogen that causes substantial economic losses in worldwide potato production. Different potato cultivars differ in resistance to PVY, from severe susceptibility, through tolerance, to complete resistance. The aim of this study was to better define the mechanisms underlying tolerant responses of potato to infection by the particularly aggressive PVY(NTN) strain. We focused on the dynamics of the primary metabolism-related processes during PVY(NTN) infection. RESULTS: A comprehensive analysis of the dynamic changes in primary metabolism was performed, which included whole transcriptome analysis, nontargeted proteomics, and photosynthetic activity measurements in potato cv. Désirée and its transgenic counterpart depleted for accumulation of salicylic acid (NahG-Désirée). Faster multiplication of virus occurred in the NahG-Désirée, with these plants developing strong disease symptoms. We show that while the dynamics of responses at the transcriptional level are extensive and bimodal, this is only partially translated to the protein level, and to the final functional outcome. Photosynthesis-related genes are transiently induced before viral multiplication is detected and it is down-regulated later on. This is reflected as a deficiency of the photosynthetic apparatus at the onset of viral multiplication only. Interestingly, specific and constant up-regulation of some RuBisCO transcripts was detected in Désirée plants, which might be important, as these proteins have been shown to interact with viral proteins. In SA-deficient and more sensitive NahG-Désirée plants, consistent down-regulation of photosynthesis-related genes was detected. A constant reduction in the photochemical efficiency from the onset of viral multiplication was identified; in nontransgenic plants this decrease was only transient. The transient reduction in net photosynthetic rate occurred in both genotypes with the same timing, and coincided with changes in stomatal conductivity. CONCLUSIONS: Down-regulation of photosynthesis-related gene expression and decreased photosynthetic activity is in line with other studies that have reported the effects of biotic stress on photosynthesis. Here, we additionally detected induction of light-reaction components in the early stages of PVY(NTN) infection of tolerant interaction. As some of these components have already been shown to interact with viral proteins, their overproduction might contribute to the absence of symptoms in cv. Désirée. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1925-2) contains supplementary material, which is available to authorized users
Genome-wide mRNA degradation is involved in bacterial adaptation
International audienc
Adaptation des bactéries à leur environnement : antagonisme entre Stabilité et Concentration des ARNm
International audienc