Disrupting the methionine biosynthetic pathway in Candida guilliermondii: characterization of the MET2 gene as counter‐selectable marker

Candida guilliermondii (teleomorph Meyerozyma guilliermondii) is an ascomycetous species belonging to the fungal CTG clade. This yeast remains actively studied as a result of its moderate clinical importance and most of all for its potential uses in biotechnology. The aim of the present study was to establish a convenient transformation system for C. guilliermondii by developing both a methionine auxotroph recipient strain and a functional MET gene as selection marker. We first disrupted the MET2 and MET15 genes encoding homoserine‐O‐acetyltransferase and O‐acetylserine O‐acetylhomoserine sulphydrylase, respectively. The met2 mutant was shown to be a methionine auxotroph in contrast to met15 which was not. Interestingly, met2 and met15 mutants formed brown colonies when cultured on lead‐containing medium, contrary to the wild‐type strain, which develop as white colonies on this medium. The MET2 wild‐type allele was successfully used to transfer a yellow fluorescent protein (YFP) gene‐expressing vector into the met2 recipient strain. In addition, we showed that the loss of the MET2‐containing YFP‐expressing plasmid can be easily observed on lead‐containing medium. The MET2 wild‐type allele, flanked by two short repeated sequences, was then used to disrupt the LYS2 gene (encoding the α‐aminoadipate reductase) in the C. guilliermondii met2 recipient strain. The resulting lys2 mutants displayed, as expected, auxotrophy for lysine. Unfortunately, all our attempts to pop‐out the MET2 marker (following the recombination of the bordering repeat sequences) from a target lys2 locus were unsuccessful using white/brown colony colour screening. Nevertheless, this MET2 transformation/disruption system represents a new versatile genetic tool for C. guilliermondii. Copyright © 2014 John Wiley & Sons, Ltd.     

Incorporation of process integration into life cycle analysis for the production of biofuels

This article presents a new approach for incorporating process-integration tools into life cycle analysis (LCA) for biofuel production. Process synthesis techniques using mass- and energy-integration tools are employed to generate various scenarios for reducing mass and energy consumption in the process. The global implications of these changes and the associated trade-offs are assessed using the LCA tool: GREET. The developed approach enables the consideration of several levels of process integration while tracking the process economics and the reduction of the net greenhouse gas emissions. Several cases of biofuels with different processing technologies have been considered in this study, and the results show that when the process-integration tools are effectively utilized and combined with LCA, better insights are obtained and the net greenhouse gas emissions decrease drastically for different biofuels.     

Review of Bioretention System Research and Design: Past, Present, and Future

This paper reviews the evolution of bioretention systems, a promising at-source storm-water best management practice. The introduction of bioretention systems in the 1990s by Prince George’s County, Md. is examined along with the motivations behind the development of the systems. A summary of the research findings on the performance of bioretention systems is provided including proposed design modifications to improve the field performance. Also included is an overview of past and current bioretention design guidelines in North America, as well as a discussion of issues surrounding the public adoption and implementation of bioretention systems. Potential alternative uses for the systems are highlighted and a review of bioretention modeling work is provided. Finally, the paper outlines research needs and anticipated future work necessary to bring about the widespread use of bioretention systems.     

An enhanced deterministic beacon advertising algorithm for building TSCH networks

In this paper, we focus first on the time needed by a node to join a Time Slotted Channel Hopping (TSCH) network, this time is called joining time. Second, we are also interested in the network building time. Since the data generated by a sensor node remain unavailable as long as this node has not yet joined the wireless sensor network, these times are of prime importance for applications having strong latency requirements on data gathering. The joining time depends on the beacon advertising policy that has been left unspecified by the standard. The contribution of this paper is triple. First, we propose an Enhanced Deterministic Beacon Advertising algorithm, called EDBA, that ensures a collision-free advertising of beacons and minimizes the average joining time. Second, we model the behavior of a joining node by a Markov chain, validated by NS3 simulations, and compute the average joining time. Third, we compare the performance of EDBA with this of MBS, considered as the best beacon advertising algorithm in the literature.     

In vivo lipolysis induced by heparin injection or fasting: a proton MR spectroscopy analysis of human plasma

The impact of induced lipolysis on the composition of plasma lipids is analyzed by proton magnetic resonance spectroscopy (MRS) in humans. The variations of the methylene and methyl resonances from lipids in lipoproteins are studied under two sets of lipolytic conditions: acute endovascular lipolysis induced by an intravenous injection of heparin and subacute lipolysis induced by short fasting. During acute lipolysis, the degradation of the very low density lipoproteins structures is well correlated to the modifications observed in the areas of CH₂ and CH₃ MRS signals. The comparison of regular spectra, spectra with water signal suppression, and spectra recorded with a spin-echo sequence provides information on the behavior of the different parts of the lipoproteins, that is, the neutral core, little affected by heparine-induced lipoprotein lipase (LPL) activation, and the surface layer supplying substrates to LPL. During 48 h of fasting, only limited modifications occur on the MR spectra, and lipolysis cannot be documented in details.Address for correspondence: CRMBM-CNRS, Faculté de Médecine, 27, bd Jean Moulin, 13005 Marseille, France.     

Porous graphitic carbon as stationary phase for LC-ICPMS separation of arsenic compounds in water

A new liquid chromatographic separation method was developed for the speciation of the four main arsenic compounds present in water. Arsenite (As(III)), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA) and arsenate (As(V)) were separated on a recently introduced stationary phase: porous graphitic carbon (PGC). The separation was first obtained under formic acid gradient conditions, but an adsorption phenomenon of As(V) on PGC was observed. To overcome this problem, As(V) was backflushed, and an efficient separation of the four solutes was achieved within 10 min. Extremely low detection limits (ranging from 10 to 70 ng x L(-1)) were obtained by coupling LC with an ICPMS. The method was successfully applied to different spiked mineral waters and a naturally arsenic-containing freshwater.