CTP synthetase activity assay by liquid chromatography tandem mass spectrometry in the multiple reaction monitoring mode

Cytidine 5′‐triphosphate synthetase (CTPS) is known to be a central enzyme in the de novo synthesis of CTP. We have recently demonstrated that a deficiency in CTPS1 is associated with an impaired capacity of activated lymphocytes to proliferate leading to a combined immunodeficiency disease. In order to better document its role in immunomodulation, we developed a method for measuring CTPS activity in human lymphocytes. Using liquid chromatography‐mass spectrometry, we quantified CTPS activity by measuring CTP in cell lysates. A stable isotope analog of CTP served as internal standard. We characterized the kinetic parameters V_max and K_m of CTPS and verified that an inhibition of the enzyme activity was induced after 3‐deazauridine (3DAU) treatment, a known inhibitor of CTPS. We then determined CTPS activity in healthy volunteers, in a family whose child displayed a homozygous mutation in CTPS1 gene and in patients who had developed or not a chronic lung allograft dysfunction (CLAD) after lung transplantation. Linearity of the CTP determination was observed up to 451 μmol/L, with accuracy in the 15% tolerance range. Michaelis‐Menten kinetics for lysates of resting cells were K_m=280±310 μmol/L for UTP, V_max=83±20 pmol/min and, for lysates of activated PBMCs, K_m=230±280 μmol/L for UTP, V_max=379±90 pmol/min. Treatment by 3DAU and homozygous mutation in CTPS1 gene abolished the induction of CTPS activity associated with cell stimulation, and CTPS activity was significantly reduced in the patients who developed CLAD. We conclude that this test is suitable to reveal the involvement of CTPS alteration in immunodeficiency.     

A generalized auxiliary equation method and its application to nonlinear Klein-Gordon and generalized nonlinear Camassa-Holm equations

With the aid of symbolic computation, a generalized auxiliary equation method is proposed to construct more general exact solutions to two types of NLPDEs. First, we present new family of solutions to a nonlinear Klein-Gordon equation, by using this auxiliary equation method including a new first-order nonlinear ODE with six-degree nonlinear term proposed by Sirendaoreji. Then, we apply an indirect F-function method very close to the F-expansion method to solve the generalized Camassa-Holm equation with fully nonlinear dispersion and fully nonlinear convection C(l,n,p). Taking advantage of the new first-order nonlinear ODE with six degree nonlinear term, this indirect F-function method is used to map the solutions of C(l,n,p) equations to those of that nonlinear ODE. As a result, we can successfully obtain in a unified way, many exact solutions.     

Supramolecular Association between γ-Cyclodextrin and Preyssler-Type Polyoxotungstate

The development of hybrid materials based on polyoxometalates constitutes a strategy for the design of multifunctional materials. The slow evaporation of an aqueous solution of [NaP₅W₃₀O₁₁₀]¹⁴⁻ in the presence of γ-Cyclodextrin (γ-CD) led to the crystallization of a K₆Na₈{[NaP₅W₃₀O₁₁₀]•(C₄₈H₈₀O₄₀)}•23H₂O (NaP₅W₃₀•1CD) supramolecular compound, which was characterized by single-crystal X-ray diffraction, IR-spectroscopy, thermogravimetric and elemental analyses. Structural analysis revealed the formation of 1:1 {[NaP₅W₃₀O₁₁₀]•[γ-CD]}¹⁴⁻ adduct in the solid state. Studies in solution by cyclic voltammetry, electrochemical impedance spectroscopy, ¹H NMR spectroscopy, and ³¹P DOSY, have demonstrated weak interactions between the inorganic anion and the macrocyclic organic molecule.     

Dynamics of a close-loop controlled MEMS resonator

The dynamics of a close-loop electrostatic MEMS resonator, proposed as a platform for ultra sensitive mass sensors, is investigated. The parameter space of the resonator actuation voltage is investigated to determine the optimal operating regions. Bifurcation diagrams of the resonator response are obtained at five different actuation voltage levels. The resonator exhibits bi-stability with two coexisting stable equilibrium points located inside a lower and an upper potential wells. Steady-state chaotic attractors develop inside each of the potential wells and around both wells. The optimal region in the parameter space for mass sensing purposes is determined. In that region, steady-state chaotic attractors develop and spend most of the time in the safe lower well while occasionally visiting the upper well. The robustness of the chaotic attractors in that region is demonstrated by studying their basins of attraction. Further, regions of large dynamic amplification are also identified in the parameter space. In these regions, the resonator can be used as an efficient long-stroke actuator.     

A sequential Monte Carlo algorithm for solving BSDEs

We present and analyze a numerical algorithm for solving BSDEs based on Picard iterations and on a sequential control variate method. Its convergence is geometric. Moreover, our algorithm provides a regular solution w.r.t. time and space. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)     

Mixed Oligoureas Based on Constrained Bicyclic and Acyclic β‐Amino Acids Derivatives: On the Significance of the Subunit Configuration for Folding

The combination of a non‐functionalized constrained bicyclo[2.2.2]octane motif along with urea linkages allowed the formation of a highly rigid 2.5_12/14 helical system both in solution and the solid state. In this work, we aimed at developing stable and functionalized systems as promising materials for biological applications in investigating the impact of this constrained motif and its configuration on homo and heterochiral mixed‐oligourea helix formation. Di‐, tetra‐, hexa‐, and octa‐oligoureas alternating the highly constrained bicyclic motif of (R) or (S) configuration with acyclic (S)‐β³‐amino acid derivatives were constructed. Circular dichroism (CD), NMR experiments, and the X‐ray crystal structure of the octamer unequivocally proved that the alternating heterochiral R/S sequences form a stable left‐handed 2.5‐helix in contrast to the mixed (S/S)‐oligoureas, which did not adopt any defined secondary structure. We observed that the (?)‐synclinal conformation around the C_α? C_β bond of the acyclic residues, although sterically less favorable than the (+)‐synclinal conformation, was imposed by the (R)‐bicyclic amino carbamoyl (BAC) residue. This highlighted the strong ability of the BAC residue to drive helical folding in heterochiral compounds. The role of the stereochemistry of the BAC unit was assessed and a model was proposed to explain the misfolding of the S/S sequences.     

Nafion as cosurfactant: solubilization of nafion in water in the presence of Pluronics

Incorporation of Nafion to aqueous solutions of Pluronics adversely impacts micellization due to extensive Nafion/copolymer interactions. Light scattering and zeta potential measurements provide evidence for the formation of sizable and stable Nafion/copolymer complexes, in expense of the neat copolymer micelles. At high copolymer concentrations, the overall interaction diagram of Nafion/copolymer reflects the competitive action of the release of packing constraints due to micellar destabilization induced by Nafion on one hand and the gelator nature of the Nafion on the other. Measurements using a quartz crystal microbalance (QCM-D) show that aqueous solutions of Pluronics (even at very low concentration) can dissolve the Nafion coating on the crystal resonator, while typical low molecular weight ionic surfactants fail to induce similar effects. These studies demonstrate that complexation with this class of copolymers is a facile route to impart dispersibility to Nafion in aqueous environments that otherwise can be achieved through tedious and harsh treatments.     

Synthesis and self-assembly of "tree-like" amphiphilic glycopolypeptides

Novel synthetic tree-like oligosaccharides-grafted-polypeptides were prepared by using Huisgen 1,3-dipolar cycloaddition between poly(γ-benzyl-L-glutamate)-block-poly(propargylglycine) and two different oligosaccharides, dextran or hyaluronan. By direct solubilisation in water, these tree-like glycopeptides spontaneously form very small assemblies with sizes below 50 nm and low polydispersity.