Velocity gradients statistics along particle trajectories in turbulent flows: the refined similarity hypothesis in the Lagrangian frame

We present an investigation of the statistics of velocity gradient related quantities, in particluar energy dissipation rate and enstrophy, along the trajectories of fluid tracers and of heavy/light particles advected by a homogeneous and isotropic turbulent flow. The Refined Similarity Hypothesis (RSH) proposed by Kolmogorov and Oboukhov in 1962 is rephrased in the Lagrangian context and then tested along the particle trajectories. The study is performed on state-of-the-art numerical data resulting from numerical simulations up to Re~400 with 2048^3 collocation points. When particles have small inertia, we show that the Lagrangian formulation of the RSH is well verified for time lags larger than the typical response time of the particle. In contrast, in the large inertia limit when the particle response time approaches the integral-time-scale of the flow, particles behave nearly ballistic, and the Eulerian formulation of RSH holds in the inertial-range.Comment: 7 pages, 7 figures; Physical Review E (accepted Dec 7, 2009

Characterization of [3H][^3H]Phenobarbital Binding to Rat Brain Membranes

The binding of $[^3H]$phenobarbital to rat brain membranes was studied in order to determine its characteristics and specificity. The binding reaction was rapid and occurred at sites of low affinity. $(K_d = 700 μM)$ and very high density $(B_{max} = 2.7 nmoll/mg protein)$. It was unaffected by temperature changes from O°C to 95°C and was maximal at pH 5. Detergents in low concentrations markedly decreased the binding, apparently without solubilizing the binding sites. It is concluded that the binding of $[^3H]$ phenobarbital is a rather non-specific interaction with the plasma membrane

Detailed design of a lattice composite fuselage structure by a mixed optimization method

In this paper, a procedure for designing a lattice fuselage barrel has been developed and it comprises three stages: first, topology optimization of an aircraft fuselage barrel has been performed with respect to weight and structural performance to obtain the conceptual design. The interpretation of the optimal result is given to demonstrate the development of this new lattice airframe concept for the fuselage barrel. Subsequently, parametric optimization of the lattice aircraft fuselage barrel has been carried out using Genetic Algorithms on metamodels generated with Genetic Programming from a 101-point optimal Latin hypercube design of experiments. The optimal design has been achieved in terms of weight savings subject to stability, global stiffness and strain requirements and then was verified by the fine mesh finite element simulation of the lattice fuselage barrel. Finally, a practical design of the composite skin complying with the aircraft industry lay-up rules has been presented. It is concluded that the mixed optimization method, combining topology optimization with the global metamodel-based approach, has allowed to solve the problem with sufficient accuracy as well as provided the designers with a wealth of information on the structural behaviour of the novel anisogrid composite fuselage design

Wind reversals in turbulent Rayleigh-Benard convection

The phenomenon of irregular cessation and subsequent reversal of the large-scale circulation in turbulent Rayleigh-B\'enard convection is theoretically analysed. The force and thermal balance on a single plume detached from the thermal boundary layer yields a set of coupled nonlinear equations, whose dynamics is related to the Lorenz equations. For Prandtl and Rayleigh numbers in the range $10^{-2} \leq \Pr \leq 10^{3}$ and 10^{7} \leq \Ra \leq 10^{12}, the model has the following features: (i) chaotic reversals may be exhibited at Ra $\geq 10^{7}$; (ii) the Reynolds number based on the root mean square velocity scales as \Re_{rms} \sim \Ra^{[0.41 ... 0.47]} (depending on Pr), and as $\Re_{rms} \sim \Pr^{-[0.66 ... 0.76]}$ (depending on Ra); and (iii) the mean reversal frequency follows an effective scaling law \omega / (\nu L^{-2}) \sim \Pr^{-(0.64 \pm 0.01)} \Ra^{0.44 \pm 0.01}. The phase diagram of the model is sketched, and the observed transitions are discussed.Comment: 4 pages, 5 figure

Periodically kicked turbulence

Periodically kicked turbulence is theoretically analyzed within a mean field theory. For large enough kicking strength A and kicking frequency f the Reynolds number grows exponentially and then runs into some saturation. The saturation level can be calculated analytically; different regimes can be observed. For large enough Re we find the saturation level to be proportional to A*f, but intermittency can modify this scaling law. We suggest an experimental realization of periodically kicked turbulence to study the different regimes we theoretically predict and thus to better understand the effect of forcing on fully developed turbulence.Comment: 4 pages, 3 figures. Phys. Rev. E., in pres

Visualization of class A GPCR oligomerization by image-based fluorescence fluctuation spectroscopy

G protein-coupled receptors (GPCRs) represent the largest class of cell surface receptors conveying extracellular information into intracellular signals. Many GPCRs have been shown to be able to oligomerize and it is firmly established that Class C GPCRs (e.g. metabotropic glutamate receptors) function as obligate dimers. However, the oligomerization capability of the larger Class A GPCRs (e.g. comprising the β-adrenergic receptors (β-ARs)) is still, despite decades of research, highly debated. Here we assess the oligomerization behavior of three prototypical Class A GPCRs, the β1-ARs, β2-ARs, and muscarinic M2Rs in single, intact cells. We combine two image correlation spectroscopy methods based on molecular brightness, i.e. the analysis of fluorescence fluctuations over space and over time, and thereby provide an assay able to robustly and precisely quantify the degree of oligomerization of GPCRs. In addition, we provide a comparison between two labelling strategies, namely C-terminally-attached fluorescent proteins and N-terminally-attached SNAP-tags, in order to rule out effects arising from potential fluorescent protein-driven oligomerization. The degree of GPCR oligomerization is expressed with respect to a set of previously reported as well as newly established monomeric or dimeric control constructs. Our data reveal that all three prototypical GPRCs studied display, under unstimulated conditions, a prevalently monomeric fingerprint. Only the β2-AR shows a slight degree of oligomerization. From a methodological point of view, our study suggests three key aspects. First, the combination of two image correlation spectroscopy methods allows addressing cells transiently expressing high concentrations of membrane receptors, far from the single molecule regime, at a density where the kinetic equilibrium should favor dimers and higher-order oligomers. Second, our methodological approach, allows to selectively target cell membrane regions devoid of artificial oligomerization hot-spots (such as vesicles). Third, our data suggest that the β1-AR appears to be a superior monomeric control than the widely used membrane protein CD86. Taken together, we suggest that our combined image correlation spectroscopy method is a powerful approach to assess the oligomerization behavior of GPCRs in intact cells at high expression levels

A universal bioluminescence resonance energy transfer sensor design enables high-sensitivity screening of GPCR activation dynamics

G-protein-coupled receptors (GPCRs) represent one of the most important classes of drug targets. The discovery of new GCPR therapeutics would greatly benefit from the development of a generalizable high-throughput assay to directly monitor their activation or de-activation. Here we screened a variety of labels inserted into the third intracellular loop and the C-terminus of the α-adrenergic receptor and used fluorescence (FRET) and bioluminescence resonance energy transfer (BRET) to monitor ligand-binding and activation dynamics. We then developed a universal intramolecular BRET receptor sensor design to quantify efficacy and potency of GPCR ligands in intact cells and real time. We demonstrate the transferability of the sensor design by cloning β-adrenergic and PTH1-receptor BRET sensors and monitored their efficacy and potency. For all biosensors, the Z factors were well above 0.5 showing the suitability of such design for microtiter plate assays. This technology will aid the identification of novel types of GPCR ligands

Development of a conformational histamine H(3) receptor biosensor for the synchronous screening of agonists and inverse agonists

The histamine H(3) receptor (H(3)R) represents a highly attractive drug target for the treatment of various central nervous system disorders, but the discovery of novel H(3)R targeting compounds relies on the assessment of highly amplified intracellular signaling events that do not only reflect H(3)R modulation and carry the risk of high false-positive and -negative screening rates. To address these limitations, we designed an intramolecular H(3)R biosensor based on the principle of bioluminescence resonance energy transfer (BRET) that reports the receptor's real-time conformational dynamics and provides an advanced tool to screen for both H(3)R agonists and inverse agonists in a live cell screening-compatible assay format. This conformational G-protein-coupled receptor (GPCR) sensor allowed us to characterize the pharmacological properties of known and new H(3) receptor ligands with unprecedented accuracy. Interestingly, we found that one newly developed H(3) receptor ligand possesses even stronger inverse agonistic activity than reference H(3)R inverse agonists including the current gold standard pitolisant. Taken together, we describe here the design and validation of the first screening-compatible H(3)R conformational biosensor that will aid in the discovery of novel H(3)R ligands and can be employed to gain deeper insights into the (in-)activation mechanism of this highly attractive drug target