Efficiency of Fish Propulsion

It is shown that the system efficiency of a self-propelled flexible body is ill-defined unless one considers the concept of quasi-propulsive efficiency, defined as the ratio of the power needed to tow a body in rigid-straight condition over the power it needs for self-propulsion, both measured for the same speed. Through examples we show that the quasi-propulsive efficiency is the only rational non-dimensional metric of the propulsive fitness of fish and fish-like mechanisms. Using two-dimensional viscous simulations and the concept of quasi-propulsive efficiency, we discuss the efficiency two-dimensional undulating foils. We show that low efficiencies, due to adverse body-propulsor hydrodynamic interactions, cannot be accounted for by the increase in friction drag

Exposure of ciprofloxacin-resistant Escherichia coli broiler isolates to subinhibitory concentrations of a quaternary ammonium compound does not increase antibiotic resistance gene transfer

Resistance to antibiotics threatens to become a worldwide health problem. An important attributing phenomenon in this context is that pathogens can acquire antibiotic resistance genes through conjugative transfer of plasmids. To prevent bacterial infections in agricultural settings, the use of veterinary hygiene products, such as disinfectants, has gained popularity and questions have been raised about their contribution to such spreading of antibiotic resistance. Therefore, this study investigated the effect of subinhibitory concentrations of benzalkoniumchloride (BKC), a quaternary ammonium compound (QAC), on the conjugative transfer of antibiotic resistance genes. Five Escherichia coli field strains originating from broiler chickens and with known transferable plasmid-mediated ciprofloxacin resistance were exposed to subinhibitory BKC concentrations: 1/3, 1/10 and 1/30 of the minimum bactericidal concentration. Antibiotic resistance transfer was assessed by liquid mating for 4 h at 25 degrees C using E. coli K12 MG1655 as recipient strain. The transfer ratio was calculated as the number of transconjugants divided by the number of recipients. Without exposure to BKC, the strains showed a ciprofloxacin resistance transfer ratio ranging from 10(-4) to 10(-7). No significant effect of exposure to subinhibitory concentrations of BKC was observed on this transfer ratio

Study of material homogeneity in the long fiber thermoset injection molding process by image texture analysis

To quantify the homogeneity of fiber dispersion in short fiber-reinforced polymer composites, a method for image texture analysis of 3-dimensional X-ray micro computed tomography (µCT) images is presented in this work. The adaption of the method to the specific requirements of the composite material is accomplished using a statistical region merging approach. Subsequently, the method is applied for evaluating the homogeneity of specimens from an intermediate step of the long fiber thermoset injection molding process as well as molded parts. This new injection molding process enables the manufacturing of parts with a flexible combination of short and long glass fibers. By using a newly developed screw element based on the Maddock mixing element design, the material homogeneity of parts molded in the long fiber injection molding process is improved

Construction and model-based analysis of a promoter library for E. coli: an indispensable tool for metabolic engineering

<p>Abstract</p> <p>Background</p> <p>Nowadays, the focus in metabolic engineering research is shifting from massive overexpression and inactivation of genes towards the model-based fine tuning of gene expression. In this context, the construction of a library of synthetic promoters of <it>Escherichia coli </it>as a useful tool for fine tuning gene expression is discussed here.</p> <p>Results</p> <p>A degenerated oligonucleotide sequence that encodes consensus sequences for <it>E. coli </it>promoters separated by spacers of random sequences has been designed and synthesized. This 57 bp long sequence contains 24 conserved, 13 semi-conserved (W, R and D) and 20 random nucleotides. This mixture of DNA fragments was cloned into a promoter probing vector (pVIK165). The ligation mixtures were transformed into competent <it>E. coli </it>MA8 and the resulting clones were screened for GFP activity by measuring the relative fluorescence units; some clones produced high fluorescence intensity, others weak fluorescence intensity. The clones cover a range of promoter activities from 21.79 RFU/OD₆₀₀ml to 7606.83 RFU/OD₆₀₀ml. 57 promoters were sequenced and used for promoter analysis. The present results conclusively show that the postulates, which link promoter strength to anomalies in the -10 box and/or -35 box, and to the length of the spacer, are not generally valid. However, by applying Partial Least Squares regression, a model describing the promoter strength was built and validated.</p> <p>Conclusion</p> <p>For <it>Escherichia coli</it>, the promoter strength can not been linked to anomalies in the -10 box and/or -35 box, and to the length of the spacer. Also a probabilistic approach to relate the promoter sequence to its strength has some drawbacks. However, by applying Partial Least Squares regression, a good correlation was found between promoter sequence and promoter strength. This PLS model can be a useful tool to rationally design a suitable promoter in order to fine tune gene expression.</p

Effect of subinhibitory exposure to quaternary ammonium compounds on the ciprofloxacin susceptibility ofEscherichia colistrains in animal husbandry

Background Quaternary ammonium compound based disinfectants are commonly used in pig and poultry husbandry to maintain farm hygiene. However, studies have shown that subinhibitory concentrations of these disinfectants may increase antibiotic resistance. Investigation of antibiotic susceptibility is usually assessed via the microbroth dilution method, although this conventional culture-based technique only provides information on the bacteriostatic activity of an antimicrobial agent. Therefore, experiments were performed to investigate the effect of prior benzalkonium chloride (BKC) exposure on the viability of subsequent ciprofloxacin (CIP) treatedEscherichia coli. Results Following CIP treatment, bacterial cell counts were significantly higher after exposure to a subinhibitory BKC concentration than without BKC exposure. The flow cytometric results suggested a BKC-dependent onset of membrane damage and loss of membrane potential. Conclusion Our results indicate a lower bactericidal effect of CIP treatment on BKC-exposedE. coliisolates compared to unexposedE. coliisolates

Transform-domain analysis of packet delay in network nodes with QoS-aware scheduling

In order to differentiate the perceived QoS between traffic classes in heterogeneous packet networks, equipment discriminates incoming packets based on their class, particularly in the way queued packets are scheduled for further transmission. We review a common stochastic modelling framework in which scheduling mechanisms can be evaluated, especially with regard to the resulting per-class delay distribution. For this, a discrete-time single-server queue is considered with two classes of packet arrivals, either delay-sensitive (1) or delay-tolerant (2). The steady-state analysis relies on the use of well-chosen supplementary variables and is mainly done in the transform domain. Secondly, we propose and analyse a new type of scheduling mechanism that allows precise control over the amount of delay differentiation between the classes. The idea is to introduce N reserved places in the queue, intended for future arrivals of class 1

Promoter knock-in: a novel rational method for the fine tuning of genes

<p>Abstract</p> <p>Background</p> <p>Metabolic engineering aims at channeling the metabolic fluxes towards a desired compound. An important strategy to achieve this is the modification of the expression level of specific genes. Several methods for the modification or the replacement of promoters have been proposed, but most of them involve time-consuming screening steps. We describe here a novel optimized method for the insertion of constitutive promoters (referred to as "promoter knock-in") whose strength can be compared with the native promoter by applying a promoter strength predictive (PSP) model.</p> <p>Results</p> <p>Our method was successfully applied to fine tune the <it>ppc </it>gene of <it>Escherichia coli</it>. While developing the promoter knock-in methodology, we showed the importance of conserving the natural leader region containing the ribosome binding site (RBS) of the gene of interest and of eliminating upstream regulatory elements (transcription factor binding sites). The gene expression was down regulated instead of up regulated when the natural RBS was not conserved and when the upstream regulatory elements were eliminated. Next, three different promoter knock-ins were created for the <it>ppc </it>gene selecting three different artificial promoters. The measured constitutive expression of the <it>ppc </it>gene in these knock-ins reflected the relative strength of the different promoters as predicted by the PSP model. The applicability of our PSP model and promoter knock-in methodology was further demonstrated by showing that the constitutivity and the relative levels of expression were independent of the genetic background (comparing wild-type and mutant <it>E. coli </it>strains). No differences were observed during scaling up from shake flask to bioreactor-scale, confirming that the obtained expression was independent of environmental conditions.</p> <p>Conclusion</p> <p>We are proposing a novel methodology for obtaining appropriate levels of expression of genes of interest, based on the prediction of the relative strength of selected synthetic promoters combined with an optimized promoter knock-in strategy. The obtained expression levels are independent of the genetic background and scale conditions. The method constitutes therefore a valuable addition to the genetic toolbox for the metabolic engineering of <it>E. coli</it>.</p

Sulforaphane rewires central metabolism to support antioxidant response and achieve glucose homeostasis

Cruciferous-rich diets, particularly broccoli, have been associated with reduced risk of developing cancers of various sites, cardiovascular disease and type-2 diabetes. Sulforaphane (SF), a sulfur-containing broccoli-derived metabolite, has been identified as the major bioactive compound mediating these health benefits. Sulforaphane is a potent dietary activator of the transcription factor Nuclear factor erythroid-like 2 (NRF2), the master regulator of antioxidant cell capacity responsible for inducing cytoprotective genes, but its role in glucose homeostasis remains unclear. In this study, we set to test the hypothesis that SF regulates glucose metabolism and ameliorates glucose overload and its resulting oxidative stress by inducing NRF2 in human hepatoma HepG2 cells. HepG2 cells were exposed to varying glucose concentrations: basal (5.5 mM) and high glucose (25 mM), in the presence of physiological concentrations of SF (10 μM). SF upregulated the expression of glutathione (GSH) biosynthetic genes and significantly increased levels of reduced GSH. Labelled glucose and glutamine experiments to measure metabolic fluxes identified that SF increased intracellular utilisation of glycine and glutamate by redirecting the latter away from the TCA cycle and increased the import of cysteine from the media, likely to support glutathione synthesis. Furthermore, SF altered pathways generating NADPH, the necessary cofactor for oxidoreductase reactions, namely pentose phosphate pathway and 1C-metabolism, leading to the redirection of glucose away from glycolysis and towards PPP and of methionine towards methylation substrates. Finally, transcriptomic and targeted metabolomics LC-MS analysis of NRF2-KD HepG2 cells generated using CRISPR-Cas9 genome editing revealed that the above metabolic effects are mediated through NRF2. These results suggest that the antioxidant properties of cruciferous diets are intricately connected to their metabolic benefits