Coupling of native IEF and extended X-ray absorption fine structure to characterize zinc-binding sites from pI isoforms of SOD1 and A4V pathogenic mutant

Extended X-ray absorption fine structure (EXAFS) has already provided high-resolution structures of metal-binding sites in a wide variety of metalloproteins. Usually, EXAFS is performed on purified metalloproteins either in solution or crystallized form but purification steps are prone to modify the metallation state of the protein. We developed a protocol to couple EXAFS analysis to metalloprotein separation using native gel electrophoresis. This coupling opens a large field of applications as metalloproteins can be characterized in their native state avoiding purification steps. Using native isoelectric focusing, the method enables the EXAFS analysis of metalloprotein pI isoforms. We applied this methodology to SOD1, wild-type, and Ala4Val mutant (A4V), a mutation found in amyotrophic lateral sclerosis (ALS) because decreased Zn affinity to SOD1 mutants is suggested to be involved in the pathogenesis of this neurodegenerative disease. We observed similar coordination structures for Zn in wild-type and mutant proteins, in all measured pI isoforms, demonstrating the feasibility of EXAFS on electrophoresis gels and suggesting that the Zn-binding site is not structurally modified in A4V SOD1 mutant.     

Lipid bilayer elasticity measurements in giant liposomes in contact with a solubilizing surfactant

A new method to probe the modification of the elasticity of phospholipid bilayers is presented. The purpose here concerns the action of a solubilizing surfactant on a vesicle bilayer. This method is based on the measure of the under-field elongation of giant magnetic-fluid-loaded liposomes. The addition of the nonionic surfactant octyl-beta-d-glucopyranoside (OG) to vesicles at sublytic levels increases the elasticity of the membrane, as shown by the value of the bending modulus K(b), which decreases. K(b) measured around 20 kT for a pure 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayer indeed reaches a few kT in the case of the mixed OG-DOPC bilayer. The purpose and interest of this study are to allow the determination of the membrane bending modulus before and after the addition of OG on the same magnetic liposome. Moreover, the experimental conditions used in this work allow the control of lipid and surfactant molar fractions in the mixed aggregates. Then, optical microscopy observation can be performed on samples in well-defined regions of the OG-phospholipid state diagram.     

Polymeric liquid crystals: cholesteric superstructure from blends of hydroxypropylcellulose esters

n-Butyric, iso-butyric, n-valeric and iso-valeric esters of hydroxypropylcellulose were synthesized. Their behaviors, in terms of cholesteric structure, glass (T_g) and anisotropic-isotropic (T_i) transition temperatures are compared to results in the literature. A 6–8 month time period from sample preparation has no influence on the cholesteric phase characteristics. Six binary blends may be formed from the four esters. They show only a T_g value at each composition, which suggests a good miscibility among the components. However, positive deviation of linearity of T_g vs. composition varies from one pair to another. Analysis of the absorption curves in UV-vis spectra show that three pairs form a single cholesteric helix containing both components while other three give a only nematic phase. Colors changing from red to violet are shown for blends of n-butyric and iso-butyric esters at different weight ratio. Copyright © 2003 John Wiley & Sons, Ltd.     

A posteriori error estimation for elliptic partial differential equations with small uncertainties

In this article, a finite element error analysis is performed on a class of linear and nonlinear elliptic problems with small uncertain input. Using a perturbation approach, the exact (random) solution is expanded up to a certain order with respect to a parameter that controls the amount of randomness in the input and discretized by finite elements. We start by studying a diffusion (linear) model problem with a random coefficient characterized via a finite number of random variables. The main focus of the article is the derivation of a priori and a posteriori error estimates of the error between the exact and approximate solution in various norms, including goal‐oriented error estimation. The analysis is then extended to a class of nonlinear problems. We finally illustrate the theoretical results through numerical examples, along with a comparison with the Stochastic Collocation method in terms of computational costs. © 2015 Wiley Periodicals, Inc. Numer Methods Partial Differential Eq 32: 175–212, 2016     

On the Identity of Some von Neumann Algebras and Some Consequences

Among von Neumann algebras, the Weyl algebra W{\mathcal{W}} generated by two unitary groups {U(α)} and {V(β)}, the algebra U{\mathcal{U}} generated by a completely non-unitary semigroup of isometries {U ₊(α)} and the Weyl algebra W₊^h{\mathcal{W}_{+}^{h}} pertaining to a semi-bounded space with homogeneous spectrum of the generator of {V(β)}, all share the property that their representations are completely reducible and the irreducible representations are equivalent. We trace this fact to the identity of these algebras, in the sense that any of them contains a representation of any of the remaining two algebras, which in turn contains the original algebra. We prove this statement by explicit construction. The aforementioned results about the representations of the algebras follow immediately from the proof for any of them. Also, by the above construction we prove for W^h₊{\mathcal{W}^{h}_{+}} the analog of a theorem by Sinai for W{\mathcal{W}} : given {V(β)} with semi-bounded homogeneous spectrum, there exists a completely non-unitary semigroup {U ₊(α)} such that {V(β)} and {U ₊(α)} generate W₊^h{\mathcal{W}_{+}^{h}}.     

Evidence of surfactant-induced formation of transient pores in lipid bilayers by using magnetic-fluid-loaded liposomes

It is often assumed that surfactant-induced permeability of lipid membranes obeys a pore-formation mechanism, but, to date, this has not been totally proven. A novel approach is developed using a magnetic fluid composed of calibrated nanocrystals of maghemite (gamma-Fe2O3) as a permeability marker. It is shown that low amounts of surfactant molecules catalyze the transient opening of unilamellar phospholipid vesicles which permit the passage of 8 nm maghemite nanospheres before closing up.     

First proficiency testing to evaluate the ability of European Union National Reference Laboratories to detect staphylococcal enterotoxins in milk products

The European Commission has designed a network of European Union-National Reference Laboratories (EU-NRLs), coordinated by a Community Reference Laboratory (CRL), for control of hygiene of milk and milk products (Council Directive 92/46/ECC). As a common contaminant of milk and milk products such as cheese, staphylococcal enterotoxins are often involved in human outbreaks and should be monitored regularly. The main tasks of the EU-CRLs were to select and transfer to the EU-NRLs a reference method for detection of enterotoxins, and to set up proficiency testing to evaluate the competency of the European laboratory network. The first interlaboratory exercise was performed on samples of freeze-dried cheese inoculated with 2 levels of staphylococcal enterotoxins (0.1 and 0.25 ng/g) and on an uninoculated control. These levels were chosen considering the EU regulation for staphylococcal enterotoxins in milk and milk products and the limit of detection of the enzyme-linked immunosorbent assay test recommended in the reference method. The trial was conducted according to the recommendations of ISO Guide 43. Results produced by laboratories were compiled and compared through statistical analysis. Except for data from 2 laboratories for the uninoculated control and cheese inoculated at 0.1 ng/g, all laboratories produced satisfactory results, showing the ability of the EU-NRL network to monitor the enterotoxin contaminant.