A rapid biosensor-based method for quantification of free and glucose-conjugated salicylic acid

<p>Abstract</p> <p>Background</p> <p>Salicylic acid (SA) is an important signalling molecule in plant defenses against biotrophic pathogens. It is also involved in several other processes such as heat production, flowering, and germination. SA exists in the plant as free SA and as an inert glucose conjugate (salicylic acid 2-<it>O</it>-β-D-glucoside or SAG). Recently, Huang <it>et al</it>. developed a bacterial biosensor that responds to free SA but not SAG, designated as <it>Acinetobacter </it>sp. ADPWH_<it>lux</it>. In this paper we describe an improved methodology for <it>Acinetobacter </it>sp. ADPWH_<it>lux</it>-based free SA quantification, enabling high-throughput analysis, and present an approach for the quantification of SAG from crude plant extracts.</p> <p>Results</p> <p>On the basis of the original biosensor-based method, we optimized extraction and quantification. SAG content was determined by treating crude extracts with β-glucosidase, then measuring the released free SA with the biosensor. β-glucosidase treatment released more SA in acetate buffer extract than in Luria-Bertani (LB) extract, while enzymatic hydrolysis in either solution released more free SA than acid hydrolysis. The biosensor-based method detected higher amounts of SA in pathogen-infected plants than did a GC/MS-based method. SA quantification of control and pathogen-treated wild-type and <it>sid2 </it>(SA induction-deficient) plants demonstrated the efficacy of the method described. Using the methods detailed here, we were able to detect as little as 0.28 μg SA/g FW. Samples typically had a standard deviation of up to 25% of the mean.</p> <p>Conclusion</p> <p>The ability of <it>Acinetobacter </it>sp. ADPWH_<it>lux </it>to detect SA in a complex mixture, combined with the enzymatic hydrolysis of SAG in crude extract, allowed the development of a simple, rapid, and inexpensive method to simultaneously measure free and glucose-conjugated SA. This approach is amenable to a high-throughput format, which would further reduce the cost and time required for biosensor-based SA quantification. Possible applications of this approach include characterization of enzymes involved in SA metabolism, analysis of temporal changes in SA levels, and isolation of mutants with aberrant SA accumulation.</p

Stories-Forward Hotel and Shopping Search Experience

When searching the internet on a phone for hotel or rental accommodations, users are inundated with multiple sources of information and options. Hotel search platforms currently provide consumers with aggregated results that require users to vertically scroll through static lists and generic property detail pages which can be overwhelming and time consuming. The new stories-forward platform described below overcomes these challenges and makes content consumption faster and more engaging by providing a visually-rich user interface experience with vertical and horizontal navigation within a story, as well as easy access to additional on-demand details without losing context

TVET in Taiwan: Preliminary Report

No abstract available

Critical Role of LuxS in the Virulence of Campylobacter jejuni in a Guinea Pig Model of Abortion

Previous studies on Campylobacter jejuni have demonstrated the role of LuxS in motility, cytolethal distending toxin production, agglutination, and intestinal colonization; however, its direct involvement in virulence has not been reported. In this study, we demonstrate a direct role of luxS in the virulence of C. jejuni in two different animal hosts. The IA3902 strain, a highly virulent sheep abortion strain recently described by our laboratory, along with its isogenic luxS mutant and luxScomplement strains, was inoculated by the oral route into both a pregnant guinea pig virulence model and a chicken colonization model. In both cases, the IA3902luxS mutant demonstrated a complete loss of ability to colonize the intestinal tract. In the pregnant model, the mutant also failed to induce abortion, while the wild-type strain was highly abortifacient. Genetic complementation of the luxSgene fully restored the virulent phenotype in both models. Interestingly, when the organism was inoculated into guinea pigs by the intraperitoneal route, no difference in virulence (abortion induction) was observed between the luxS mutant and the wild-type strain, suggesting that the defect in virulence following oral inoculation is likely associated with a defect in colonization and/or translocation of the organism out of the intestine. These studies provide the first direct evidence that LuxS plays an important role in the virulence of C. jejuni using an in vivomodel of natural disease

miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family

Nicotine binds to and activates a family of ligand-gated ion channels, neuronal nicotinic acetylcholine receptors (nAChRs). Chronic nicotine exposure alters the expression of various nAChR subtypes, which likely contributes to nicotine dependence; however, the underlying mechanisms regulating these changes remain unclear. A growing body of evidence indicates that microRNAs (miRNAs) may be involved in nAChR regulation. Using bioinformatics, miRNA library screening, site-directed mutagenesis, and gene expression analysis, we have identified a limited number of miRNAs that functionally interact with the 3\u27-untranslated regions (3\u27 UTRs) of mammalian neuronal nAChR subunit genes. In silico analyses revealed specific, evolutionarily conserved sites within the 3\u27 UTRs through which the miRNAs regulate gene expression. Mutating these sites disrupted miRNA regulation confirming the in silico predictions. In addition, the miRNAs that target nAChR 3\u27 UTRs are expressed in mouse brain and are regulated by chronic nicotine exposure. Furthermore, we show that expression of one of these miRNAs, miR-542-3p, is modulated by nicotine within the mesocorticolimbic reward pathway. Importantly, overexpression of miR-542-3p led to a decrease in the protein levels of its target, the nAChR beta2 subunit. Bioinformatic analysis suggests that a number of the miRNAs play a general role in regulating cholinergic signaling. Our results provide evidence for a novel mode of nicotine-mediated regulation of the mammalian nAChR gene family

miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family

Nicotine binds to and activates a family of ligand-gated ion channels, neuronal nicotinic acetylcholine receptors (nAChRs). Chronic nicotine exposure alters the expression of various nAChR subtypes, which likely contributes to nicotine dependence; however, the underlying mechanisms regulating these changes remain unclear. A growing body of evidence indicates that microRNAs (miRNAs) may be involved in nAChR regulation. Using bioinformatics, miRNA library screening, site-directed mutagenesis, and gene expression analysis, we have identified a limited number of miRNAs that functionally interact with the 3\u27-untranslated regions (3\u27 UTRs) of mammalian neuronal nAChR subunit genes. In silico analyses revealed specific, evolutionarily conserved sites within the 3\u27 UTRs through which the miRNAs regulate gene expression. Mutating these sites disrupted miRNA regulation confirming the in silico predictions. In addition, the miRNAs that target nAChR 3\u27 UTRs are expressed in mouse brain and are regulated by chronic nicotine exposure. Furthermore, we show that expression of one of these miRNAs, miR-542-3p, is modulated by nicotine within the mesocorticolimbic reward pathway. Importantly, overexpression of miR-542-3p led to a decrease in the protein levels of its target, the nAChR beta2 subunit. Bioinformatic analysis suggests that a number of the miRNAs play a general role in regulating cholinergic signaling. Our results provide evidence for a novel mode of nicotine-mediated regulation of the mammalian nAChR gene family

Theorising the Role of Public Vocational Education Institutions Using the Capabilities Approach

This paper observes several limitations of human capital theory, both as a description of the way qualifications are used in the labour market, and in severely limiting the potential roles vocational education. It proposes as an alternative the human capabilities approach which posits that the goal should be for everyone to have the capability to be and do what they have reason to value. The paper reports the application of human capabilities as productive capabilities which are located in and concentrate on an intermediate specialised level, the vocational stream which links occupations that share common practices, knowledge, skills and personal attributes. The paper reports an application of the concept of productive capabilities to seven countries: Argentina, Australia, Côte d’Ivoire, England, Ethiopia, Germany, South Africa and Taiwan. From this the report finds that productive capabilities rest upon broader social, economic, cultural, and physical resources.

Small Odd Prime Field Multivariate PKCs

We show that Multivariate Public Key Cryptosystems (MPKCs) over fields of small odd prime characteristic, say 31, can be highly efficient. Indeed, at the same design security of $2^{80}$ under the best known attacks, odd-char MPKC is generally faster than prior MPKCs over \GF{2^k}, which are in turn faster than ``traditional\u27\u27 alternatives. This seemingly counter-intuitive feat is accomplished by exploiting the comparative over-abundance of small integer arithmetic resources in commodity hardware, here embodied by SSE2 or more advanced special multimedia instructions on modern x86-compatible CPUs. We explain our implementation techniques and design choices in implementing our chosen MPKC instances modulo small a odd prime. The same techniques are also applicable in modern FPGAs which often contains a large number of multipliers