A Disulfide Bridge Allows for Site‐Selective Binding in Liver Bile Acid Binding Protein Thereby Stabilising the Orientation of Key Amino Acid Side Chains

The presence of a disulfide bridge in liver bile acid binding protein (L‐BABP/S‐S) allows for site‐selective binding of two bile acids, glycochenodeoxycholic (GCDA) and glycocholic acid (GCA), differing only in the presence of a hydroxyl group. The protein form devoid of the disulfide bridge (L‐BABP) binds both bile salts without discriminating ability. We investigate the determinants of the molecular recognition process in the formation of the heterotypic L‐BABP/S‐S complex with GCA and GCDA located in the superficial and inner protein sites, respectively. The comparison of the NMR spectroscopy structure of heterotypic holo L‐BABP/S‐S, the first reported for this protein family, with that of the homotypic L‐BABP complex demonstrates that the introduction of a S–S link between adjacent strands changes the conformation of three key residues, which function as hot‐spot mediators of molecular discrimination. The favoured χ₁ rotameric states (t, g⁺ and g^? for E99, Q100 and E109 residues, respectively) allow the onset of an extended intramolecular hydrogen‐bond network and the consequent stabilisation of the side‐chain orientation of a buried histidine, which is capable of anchoring a specific ligand.     

Plant Oil Bodies: Novel Carriers to Deliver Lipophilic Molecules

Oil bodies (OBs) are specialised organelles ubiquitously detected in plant oil seeds, which serve as lipid storage compartments. OBs consist of a hydrophobic core of triacylglycerol (TAGs), surrounded by a monolayer of phospholipids (PLs) embedded with some specific proteins with a size ranging from 0.5 to 2 μm. In this work, we report an easy method to reconstitute OBs starting from their constituents and to encapsulate lipophilic molecules, i.e. the fluorescent fluorescein isothiocyanate (FITC) and carboxyfluorescein (CF), into reconstituted OBs. This methods allowed us to produce OBs 4- to 10-fold smaller (50–200 nm) than the native one and to obtain a good recovery (about 40%) of both the fluorescent compounds used in the present work. The properties of reconstituted OBs were investigated by a combination of Brewster angle microscopy, scanning force microscopy, ζ-potential techniques. OBs were stable and formed ordered monolayers when patterned on hydrophobic substrates whereas they showed a higher tendency to aggregate into larger, coalescing OBs when were deposited onto hydrophilic substrates or at the air/water interface. Furthermore, we verified the uptake of FITC-loaded OBs by the MCF-7 breast cancer cell line. Our results indicated that OBs could be envisaged as novel carriers to deliver hydrophobic bioactive compounds.     

Symmetry energy from elliptic flow in Au + Au

The elliptic-flow ratio of neutrons with respect to protons or light complex particles in reactions of neutron-rich systems at relativistic energies is proposed as an observable sensitive to the strength of the symmetry term in the equation of state at supra-normal densities. The results obtained from the existing FOPI/LAND data for ¹⁹⁷Au + ¹⁹⁷Au collisions at 400 MeV/nucleon in comparison with the UrQMD model favor a moderately soft symmetry term with a density dependence of the potential term proportional to γ(ρ/ρ₀)${(\rho /{\rho }_0)}^{\gamma }$ with γ=0.9±0.4$\gamma =0.9\pm 0.4$.     

Use of design of experiments to optimize high-throughput semipreparative LC and LC/MS methods

This paper describes an investigation into decreasing the run time for high-throughput semipreparative RP-HPLC methods without compromising the resolution. Experimental design was used to devise a small set of experiments in which factors, including solvent flow rate, solvent/column temperature, at-column dilution, and run time were varied systematically. The results were analyzed by means of multiple regression and partial least squares to generate a model relating the factors to the results, showing which factors are important. The model was then used to determine the optimal conditions.     

A new time-domain frequency-selective quantification algorithm

In this paper a new time-domain frequency-selective quantification algorithm is presented. Frequency-selective quantification refers to a method that analyzes spectral components in a selected frequency region, ignoring all the other components outside. The algorithm, referred to as MeFreS (Metropolis Frequency-Selective), is based on rank minimization of an opportune Hankel matrix. The minimization procedure is satisfied by the down-hill simplex method, implemented with the simulated annealing method. MeFreS does not use any preprocessing step or filter to suppress nuisance peaks, but the signal model function is directly fitted. In this manner, neither inherent signal distortions nor estimation biases to be corrected occur. The algorithm was tested with Monte Carlo simulations. A comparison with VARPRO and AMARESw algorithms was carried out. Finally, two samples of known content from NMR data were quantified.     

Quadratic programming with M-Matrices

In this paper, we study the problem of quadratic programming with M-matrices. We describe (1) an effective algorithm for the case where the variables are subject to a lower-bound constraint, and (2) an analogous algorithm for the case where the variables are subject to lower-and-upper-bound constraints. We demonstrate the special monotone behavior of the iterate and gradient vectors. The result on the gradient vector is new. It leads us to consider a simple updating procedure which preserves the monotonicity of both vectors. The procedures uses the fact that an M-matrix has a nonnegative inverse. Two new algorithms are then constructed by incorporating this updating procedure into the two given algorithms. We give numerical examples which show that the new methods can be more efficient than the original ones.