Detection of quorum sensing molecules in Burkholderia cepacia culture supernatants with enzyme-linked immunosorbent assays

The Burkholderia cepacia complex (Bcc) employs a quorum sensing (QS) mechanism which is a cell density-dependent bacterial communication system to regulate certain gene expressions. As with many other Gram-negative bacteria, Burkholderia cepacia species use (N-acyl-)homoserine lactones (AHLs or HSLs) as signalling molecules. Because of the essential role of QS in bacterial behavior, the aim of this study was to demonstrate the applicability of our in-house-developed enzyme-linked immunosorbent assays (ELISAs) for the detection of bacterial activities via HSLs in B. cepacia strain LA3 culture supernatants. For this purpose the previously developed monoclonal antibodies (mAbs) HSL1/2-2C10 and HSL1/2-4H5 were exploited. N-3-Oxo-decanoyl-L-homoserine lactone (3-oxo-C10-HSL) was used as main analyte throughout all experiments. With the bacterial culture medium (named ABC medium) a matrix effect in both ELISAs was visible (slight increase in optical density, shift in test midpoints (IC₅₀) and working ranges). For example, ELISA with mAb HSL1/2-2C10 and enzyme tracer HSL3-HRP (HSL derivative conjugated to horseradish peroxidase) had an IC₅₀ of 120 μg L⁻¹ for 3-oxo-C10-HSL in phosphate-buffered saline versus 372 μg L⁻¹ in ABC medium. A significant increase of HSLs in B. cepacia strain LA3 culture supernatants after 12 h to 48 h of growth was observed. Although the analytical result of these immunoassays cannot distinguish HSLs from homoserines (HSs), the appearance of these compounds can be easily followed. Hydrolysis and spiking experiments were carried out with these biological samples. According to our knowledge, these are the first immunoassays for the detection of quorum sensing molecules in biological culture supernatants. This study provides a cost-effective, fast, and sensitive analytical method for detection of HSLs/HSs in biological samples without complex sample preparation and will offer a quick idea about B. cepacia activities. The low sample amount requirement (less than 1 mL) constitutes a tremendous advantage for many analytical questions with biological samples.     

Search for sub-eV mass solar axions by the CERN Axion Solar Telescope with 3He buffer gas

The CERN Axion Solar Telescope (CAST) has extended its search for solar axions by using (3)He as a buffer gas. At T=1.8 K this allows for larger pressure settings and hence sensitivity to higher axion masses than our previous measurements with (4)He. With about 1 h of data taking at each of 252 different pressure settings we have scanned the axion mass range 0.39 eV?m(a)?0.64 eV. From the absence of excess x rays when the magnet was pointing to the Sun we set a typical upper limit on the axion-photon coupling of g(aγ)?2.3×10(-10) GeV(-1) at 95% C.L., the exact value depending on the pressure setting. Kim-Shifman-Vainshtein-Zakharov axions are excluded at the upper end of our mass range, the first time ever for any solar axion search. In the future we will extend our search to m(a)?1.15 eV, comfortably overlapping with cosmological hot dark matter bounds.     

High precision study of muon catalyzed fusion in D2 and HD gas

Muon catalyzed dd fusion in D₂ and HD gases in the temperature range from 28 to 350 K was investigated in a series of experiments based on a time-projection ionization chamber operating with pure hydrogen. All main observables in this reaction chain were measured with high absolute precision including the resonant and non-resonant ddμ formation rates, the rate for hyperfine transitions in dμ atoms, the branching ratio of the two charge symmetric fusion channels ³He + n and t + p and the muon sticking probability. The report presents the final analysis of the data together with a comprehensive comparison with calculations based on recent μCF theories. The energy of the loosely bound ddμ state with quantum numbers J = 1, ν = 1, which is central to the mechanism of resonant molecule formation, is extracted with precision ?₁₁(fit) = ?1.9651(7) eV. in impressive agreement with the latest theoretical results ?₁₁(theory) = ?1.9646 eV.     

Ambient space formulations and statistical mechanics of holonomically constrained Langevin systems

The most classic approach to the dynamics of an n-dimensional mechanical system constrained by d independent holonomic constraints is to pick explicitly a new set of (n − d) curvilinear coordinatesparametrizingthe manifold of configurations satisfying the constraints, and to compute the Lagrangian generating the unconstrained dynamics in these (n − d) configuration coordinates. Starting from this Lagrangian an unconstrained Hamiltonian H(q,p) on 2(n−d) dimensional phase space can then typically be defined in the standard way via a Legendre transform. Furthermore, if the system is in contact with a heat bath, the associated Langevin and Fokker-Planck equations can be introduced. Provided that an appropriate fluctuation-dissipation condition is satisfied, there will be a canonical equilibrium distribution of the Gibbs form exp(−βH) with respect to the flat measure dqdp in these 2(n − d) dimensional curvilinear phase space coordinates. The existence of (n − d) coordinates satisfying the constraints is often guaranteed locally by an implicit function theorem. Nevertheless in many examples these coordinates cannot be constructed in any tractable form, even locally, so that other approaches are of interest. In ambient space formulations the dynamics are defined in the full original n-dimensional configuration space, and associated 2n-dimensional phase space, with some version of Lagrange multipliers introduced so that the 2(n − d) dimensional sub-manifold of phase space implied by the holonomic constraints and their time derivative, is invariant under the dynamics. In this article we review ambient space formulations, and explain that for constrained dynamics there is in fact considerable freedom in how a Hamiltonian form of the dynamics can be constructed. We then discuss and contrast the Langevin and Fokker-Planck equations and their equilibrium distributions for the different forms of ambient space dynamics.     

On two possible definitions of the free energy for collective variables

The aim of this mini-review article is to clarify the relation between two distinct formulations of the thermodynamic free energy for collective variables which can be found in the molecular dynamics literature. In doing so, we discuss the different ensemble concepts underlying the two definitions and reveal their relation to strong confinement (restraints) and molecular constraints. The latter analysis is based on a variant of Federer’s coarea formula which can be regarded as a generalization of Fubini’s theorem for iterated integrals to curvilinear coordinates and which implies the famous “blue moon” ensemble identity for computing conditional expectations using constrained simulations. For illustration we will present a few paradigmatic examples.     

Deep drawing of a layered composite: material characterization and finite-element simulation

Composite materials are widely used in different industrial fields, because of their good formability and their high strength to weight ratio. In the present work a triple-layered sandwich composite is investigated. Experimental tests at room temperature are carried out for the materials constituting the composite. A finite element model of a deep-drawing process of the composite is performed, where a finite strain constitutive model for the metal part, with material parameters calibrated to uniaxial tensile tests, has been implemented. Experimental results are compared to the numerical simulations in view of validation purposes. (© 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim)     

Powder compaction. Experiments,modeling and simulation

Field-assisted sintering technology (FAST) is a combined thermal and mechanical loading process to compact and sinter a powder material within one process step. In this short essay a constitutive model of thermo-viscoplasticity is proposed representing most of the phenomena observed in the experiments. The constitutive model is calibrated to the experimental data and some predicted experiments are compared with constitutive model showing appropriate results. (© 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim)     

Project Scheduling with Multiple Modes: A Genetic Algorithm

In this paper we consider the resource-constrained project scheduling problem with multiple execution modes for each activity and makespan minimization as objective. We present a new genetic algorithm approach to solve this problem. The genetic encoding is based on a precedence feasible list of activities and a mode assignment. After defining the related crossover, mutation, and selection operators, we describe a local search extension which is employed to improve the schedules found by the basic genetic algorithm. Finally, we present the results of our thorough computational study. We determine the best among several different variants of our genetic algorithm and compare it to four other heuristics that have recently been proposed in the literature. The results that have been obtained using a standard set of instances show that the new genetic algorithm outperforms the other heuristic procedures with regard to a lower average deviation from the optimal makespan.     

Patching over fields

We develop a new form of patching that is both far-reaching and more elementary than the previous versions that have been used in inverse Galois theory for function fields of curves. A key point of our approach is to work with fields and vector spaces, rather than rings and modules. After presenting a self-contained development of this form of patching, we obtain applications to other structures such as Brauer groups and differential modules.