New Protocol Based on UHPLC-MS/MS for Quantitation of Metabolites in Xylose-Fermenting Yeasts

Xylose fermentation is a bottleneck in second-generation ethanol production. As such, a comprehensive understanding of xylose metabolism in naturally xylose-fermenting yeasts is essential for prospection and construction of recombinant yeast strains. The objective of the current study was to establish a reliable metabolomics protocol for quantification of key metabolites of xylose catabolism pathways in yeast, and to apply this protocol to Spathaspora arborariae. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to quantify metabolites, and afterwards, sample preparation was optimized to examine yeast intracellular metabolites. S. arborariae was cultivated using xylose as a carbon source under aerobic and oxygen-limited conditions. Ion pair chromatography (IPC) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) were shown to efficiently quantify 14 and 5 metabolites, respectively, in a more rapid chromatographic protocol than previously described. Thirteen and eleven metabolites were quantified in S. arborariae under aerobic and oxygen-limited conditions, respectively. This targeted metabolomics protocol is shown here to quantify a total of 19 metabolites, including sugars, phosphates, coenzymes, monosaccharides, and alcohols, from xylose catabolism pathways (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) in yeast. Furthermore, to our knowledge, this is the first time that intracellular metabolites have been quantified in S. arborariae after xylose consumption. The results indicated that fine control of oxygen levels during fermentation is necessary to optimize ethanol production by S. arborariae. The protocol presented here may be applied to other yeast species and could support yeast genetic engineering to improve second generation ethanol production.

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Reductive Cross‐Coupling Reactions between Two Electrophiles

Reductive cross‐electrophile coupling reactions have recently been developed to a versatile and sustainable synthetic tool for selective C?C bond formation. The employment of cheap and abundant electrophiles avoids the pre‐formation and handling of organometallic reagents. In situ reductive coupling is effected in the presence of a transition‐metal catalyst (Ni, Co, Pd, Fe) and a suitable metallic reductant (Mn, Zn, Mg). This Concept article assesses the current state of the art and summarizes recent protocols with various combinations of alkyl, alkenyl, allyl, and aryl reagents and highlights key mechanistic studies.     

Dépose dynamique d'un micro-objet saisi par adhésionManipulation of micro-objects using adhesion forces and dynamical effects

This paper describes a dynamical strategy for releasing micro objects picked-up by means of adhesion forces. While sticking effects are used in order to capture an object by adequately choosing a high surface energy constitutive material for the end-effector, these same effects handicap considerably the release. We propose to take advantage of the inertial effects of both the end-effector and the manipulated object to overbalance adhesion forces and to achieve the release. Simulations show that for this purpose, accelerations as high as 10⁵ m/s² are needed. Successful manipulation of a 40 μm radius glass sphere is demonstrated. To cite this article: S. Haliyo et al., C. R. Mecanique 331 (2003).     

Improvement of the ESR detection of irradiated food containing cellulose employing a simple extraction method

Fruit may be irradiated at rather low doses, below 1 kGy in combination treatments or for quarantine purposes. To improve the ESR detection sensitivity of irradiated fruit de Jesus et al. (Int. J. Food Sci. Technol. 34 (1999) 173.) proposed extracting the fruit pulp with 80% ethanol and measuring the residue with ESR using low power (0.25 mW) for detection of ‘cellulosic’ radicals. An improvement in ESR sensitivity using the extraction procedure could be confirmed in this paper for strawberries and papayas. In most cases, a radiation dose of 0.5 kGy could be detected in both fruits even after 2–3 weeks storage. In addition, some herbs and spices were also tested, but only for a few of them the ESR detection of the ‘cellulosic’ signal was improved by previous alcoholic extraction.

As an alternative to ESR measurements, other detection methods like DNA Comet Assay and thermoluminescence were also tested.     

Pyrrolidinium Ionic Liquid Crystals

N‐Alkyl‐N‐methylpyrrolidinium cations have been used for the design of ionic liquid crystals, including a new type of uranium‐containing metallomesogen. Pyrrolidinium salts with bromide, bis(trifluoromethylsulfonyl)imide, tetrafluoroborate, hexafluorophosphate, thiocyanate, tetrakis(2‐ thenoyltrifluoroacetonato)europate(III) and tetrabromouranyl counteranions were prepared. For the bromide salts and tetrabromouranyl compounds, the chain length of the alkyl group C_nH_2n+1 was varied from eight to twenty carbon atoms (n=8, 10–20). The compounds show rich mesomorphic behaviour: highly ordered smectic phases (the crystal smectic E phase and the uncommon crystal smectic T phase), smectic A phases, and hexagonal columnar phases were observed, depending on chain length and anion. This work gives better insight into the nature and formation of the crystal smectic T phase, and the molecular requirements for the appearance of this highly ordered phase. This uncommon tetragonal mesophase is thoroughly discussed on the basis of detailed powder X‐ray diffraction experiments and in relation to the existing literature. Structural models are proposed for self‐assembly of the molecules within the smectic layers. In addition, the photophysical properties of the compounds containing a metal complex anion were investigated. For the uranium‐containing mesogens, luminescence can be induced by dissolving them in an ionic liquid matrix. The europium‐containing compound shows intense red photoluminescence with high colour purity.     

Replication technology for optical microsystems

Replication technology is playing an increasingly important role in the production of optical microsystems and micro-optical elements. Hot embossing, injection moulding and UV-embossing all can produce high-quality optical elements in very cost-effective processes. New sol–gel materials allow the combination of replication with lithography to leave selected areas material-free for sawing and bonding. The development of wafer-scale replication technology using UV-curable sol–gel and polymer materials enables refractive and diffractive micro-optical elements as well as micro-mechanical alignment features to be replicated directly onto glass substrates or onto semiconductor device wafers. Grating nanostructures with linewidths less than 100 nm have been replicated into polymer and sol–gel materials for the cost-effective fabrication of large area subwavelength structures for applications such as polarisers and buried grating security features.     

High resolution Godunov‐type schemes with small stencils

Higher‐order Godunov‐type schemes have to cope with the following two problems: (i) the increase in the size of the stencil that make the scheme computationally expensive, and (ii) the monotony‐preserving treatments (limiters) that must be implemented to avoid oscillations, leading to strong damping of the solution, in particular linear waves (e.g. acoustic waves). When too compressive, limiting procedures may also trigger the instability of oscillatory numerical solutions (e.g. in advection–dispersion phenomena) via the artificial amplification of the shorter modes. The present paper proposes a new approach to carry out the reconstruction. In this approach, the values of the flow variable at the edges of the computational cells are obtained directly from the reconstruction within these cells. This method is applied to the MUSCL and DPM schemes for the solution of the linear advection equation. The modified DPM scheme can capture contact discontinuities within one computational cell, even after millions of time steps at Courant numbers ranging from 1 to values as low as 10^‐4. Linear waves are subject to negligible damping. Application of the method to the DPM for one‐dimensional advection–dispersion problems shows that the numerical instability of oscillatory solutions caused by the over compressive, original DPM limiter is eliminated. One‐ and two‐dimensional shallow water simulations show an improvement over classical methods, in particular for two‐dimensional problems with strongly distorted meshes. The quality of the computational solution in the two‐dimensional case remains acceptable even for mesh aspect ratios Δx/Δy as large as 10. The method can be extend to the discretization of higher‐order PDEs, allowing third‐order space derivatives to be discretized using only two cells in space. Copyright © 2004 John Wiley & Sons, Ltd.