APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel Variants

Hypobetalipoproteinemia is characterized by LDL-cholesterol and apolipoprotein B (apoB) plasma levels below the fifth percentile for age and sex. Familial hypobetalipoproteinemia (FHBL) is mostly caused by premature termination codons in the APOB gene, a condition associated with fatty liver and steatohepatitis. Nevertheless, many families with a FHBL phenotype carry APOB missense variants of uncertain significance (VUS). We here aimed to develop a proof-of-principle experiment to assess the pathogenicity of VUS using the genome editing of human liver cells. We identified a novel heterozygous APOB-VUS (p.Leu351Arg), in a FHBL family. We generated APOB knock-out (KO) and APOB-p.Leu351Arg knock-in Huh7 cells using CRISPR-Cas9 technology and studied the APOB expression, synthesis and secretion by digital droplet PCR and ELISA quantification. The APOB expression was decreased by 70% in the heterozygous APOB-KO cells and almost abolished in the homozygous-KO cells, with a consistent decrease in apoB production and secretion. The APOB-p.Leu351Arg homozygous cells presented with a 40% decreased APOB expression and undetectable apoB levels in cellular extracts and supernatant. Thus, the p.Leu351Arg affected the apoB secretion, which led us to classify this new variant as likely pathogenic and to set up a hepatic follow-up in this family. Therefore, the functional assessment of APOB-missense variants, using gene-editing technologies, will lead to improvements in the molecular diagnosis of FHBL and the personalized follow-up of these patients.     

Long Distance Enhancement of Nonlinear Optical Properties Using Low Concentration of Plasmonic Nanostructures in Dye Doped Monolithic Sol–Gel Materials

Monolithic sol–gel silica composites incorporating platinum‐based chromophores and various types of gold nanoparticles (AuNPs) are prepared and polished to high optical quality. Their photophysical properties are investigated. The glass materials show well‐defined localized surface plasmon resonance (SPR) absorbance from the visible to NIR. No redshifts of the AuNP plasmon absorption peaks due to the increase in nanoparticle doping concentration are observed in the glasses, proving that no or very small SPR coupling effects occur between the AuNPs. At 600 nm excitation, but not at 532 nm, the AuNPs improve the nonlinear absorption performance of glasses codoped with 50 × 10^?3 m of a Pt‐acetylide chromophore. The glasses doped with lower concentrations of AuNPs (2–5 μm average distance) and 50 × 10^?3 m in chromophore, show a marked improvement in nonlinear absorption, with no or only small improvement for the more highly AuNP doped glasses. This study shows the importance of excitation wavelength and nanoparticle concentration for composite systems employing AuNPs to improve two‐photon absorption of chromophores.     

Single‐Step Deposition of Au‐ and Pt‐Nanoparticle‐Functionalized Tungsten Oxide Nanoneedles Synthesized Via Aerosol‐Assisted CVD,and Used for Fabrication of Selective Gas Microsensor Arrays

Tungsten oxide nanostructures functionalized with gold or platinum NPs are synthesized and integrated, using a single‐step method via aerosol‐assisted chemical vapour deposition, onto micro‐electromechanical system (MEMS)‐based gas‐sensor platforms. This co‐deposition method is demonstrated to be an effective route to incorporate metal nanoparticles (NP) or combinations of metal NPs into nanostructured materials, resulting in an attractive way of tuning functionality in metal oxides (MOX). The results show variations in electronic and sensing properties of tungsten oxide according to the metal NPs introduced, which are used to discriminate effectively analytes (C₂H₅OH, H₂, and CO) that are present in proton‐exchange fuel cells. Improved sensing characteristics, in particular to H₂, are observed at 250 °C with Pt‐functionalized tungsten oxide films, whereas non‐functionalized tungsten oxide films show responses to low concentrations of CO at low temperatures. Differences in the sensing characteristics of these films are attributed to the different reactivities of metal NPs (Au and Pt), and to the degree of electronic interaction at the MOX/metal NP interface. The method presented in this work has advantages over other methods of integrating nanomaterials and devices, of having fewer processing steps, relatively low processing temperature, and no requirement for substrate pre‐treatment.     

The Performance of Independent Active SMEs in French Competitiveness Clusters

This article analyses the evolution of independent members of French independent competitiveness clusters. It compares small and medium-sized enterprises (SMEs) that participated in collaborative research projects, funded by French competitiveness clusters, against similar control companies that were not part of such projects and not funded by the policy, over the period 2005–2013. French competitiveness clusters seek to enhance SMEs’ size, performance, innovation and employment; the current study therefore assesses their outcomes in terms of capital invested and equity returns. The findings from a difference-in-differences analysis reveal that sales, employment, R&D investments, and value added all increase in relative terms, but financial returns exhibit significant relative decreases for the smallest companies, whereas larger firms record strong increases. The inability of the smallest SMEs to reap gains from their cluster membership should be a matter of great concern to both companies and policy-makers.     

5-Axis tool path smoothing based on drive constraints

In high speed machining, the real feedrate is often lower than the programmed one. This reduction of the feedrate is mainly due to the physical limits of the drives, and affects machining time as well as the quality of the machined surface. Indeed, if the tool path presents sharp geometrical variations the feedrate has to be decreased to respect the drive constraints in terms of velocity, acceleration and jerk. Thus, the aim of this paper is to smooth 5-axis tool paths in order to maximize the real feedrate and to reduce the machining time.Velocity, acceleration and jerk limits of each drive allow to compute an evaluation of the maximum reachable feedrate which is then used to localize the areas where the tool path has to be smoothed. So starting from a given tool path, the proposed algorithm iteratively smoothes the joint motions in order to raise the real feedrate. This algorithm has been tested in 5-axis end milling of an airfoil and in flank milling of an impeller for which a N-buffer algorithm is used to control the geometrical deviations. An important reduction of the measured machining time is demonstrated in both examples.     

Effects of nematic coupling on the phase behaviour of nematogen mixtures

The phase behaviour of binary nematogen mixtures of side‐chain liquid crystal crosslinked polymers and low molecular weight liquid crystals is investigated with particular emphasis on the effects of nematic coupling. The cross nematic quadrupole parameter ν₁₂ is assumed to be proportional to the geometric average of ν₁₁ and ν₂₂ characteristic of single nematogens. In the weak coupling limit, the proportionality constant is lower than 1, and the phase diagram exhibits a reduced miscibility of the nematogens. In the case of strong coupling, the proportionality constant exceeds 1 resulting in higher miscibility. This is characterized by a nematic order that extends to temperatures above the upper nematic–isotropic transition temperature. A wide region of miscibility emerges showing a single nematic phase. Nematogens having similar nematic–isotropic transition temperatures exhibit different phase properties from systems with widely separated transition temperatures. Effects of the polymer volume fraction at crosslinking, rubber elasticity parameters of the network, and the Flory–Huggins interaction parameter on the equilibrium phase diagram of these systems are discussed. © 2001 Society of Chemical Industry     

Study of the effects of structural uncertainties on a fractional system of the first kind – application in vibration isolation with the CRONE suspension

In this paper, we deal with the effects of the uncertainties on a fractional system of the first kind, mainly on the frequency-domain and the time-domain responses. For the structural uncertainties, two main aspects are studied: the nonlinearities of the physical components used to realize the fractional system and the consideration of the previously neglected dynamics of the system. Both uncertainties are introduced for the hydropneumatic CRONE suspension, previously synthesized and realized without taking into consideration these uncertainties. So, the novel approach treated in this work is to find whether the uncertainties, which were previously neglected in the synthesis and the realization phases, alter the behaviour of the system or not. The results show that the fractional order system behaviour is not affected.     

Immobilised yeast grape must deacidification in a recycle fixed bed reactor

Maloalcoholic fermentation (MAF) of grape must by Schizosaccharomyces pombe immobilised in calcium‐alginate double‐layer beads (ProMalic^®) was studied in Erlenmeyer flasks and in a total recycle fixed‐bed reactor operating in batch mode. The reaction is pseudo‐first order with respect to l ‐malic acid and under similar conditions deacidification is faster in the recycle reactor. This was attributed to mass transfer limitations which were confirmed in the recycle reactor by studying the influence of yeast load on the rate of MAF. Mass transfer limitations are also responsible for the lower activation energy of fermentation with the immobilised yeast (67 ± 9 kJ mol^?1) in comparison with the free cells (126 ± 19 kJ mol^?1). Alcoholic fermentation and MAF were performed simultaneously, both in the recycle reactor and in the industrial trials, confirming the efficacy of immobilised S. pombe to reduce grape must acidity without interfering with the main fermentation. Altogether, the present results are useful for the scale‐up of a recycle reactor to process large volumes of grape must.