The enhanced cyan fluorescent protein: a sensitive pH sensor for fluorescence lifetime imaging

pH is an important parameter that affects many functions of live cells, from protein structure or function to several crucial steps of their metabolism. Genetically encoded pH sensors based on pH-sensitive fluorescent proteins have been developed and used to monitor the pH of intracellular compartments. The quantitative analysis of pH variations can be performed either by ratiometric or fluorescence lifetime detection. However, most available genetically encoded pH sensors are based on green and yellow fluorescent proteins and are not compatible with multicolor approaches. Taking advantage of the strong pH sensitivity of enhanced cyan fluorescent protein (ECFP), we demonstrate here its suitability as a sensitive pH sensor using fluorescence lifetime imaging. The intracellular ECFP lifetime undergoes large changes (32 %) in the pH 5 to pH 7 range, which allows accurate pH measurements to better than 0.2 pH units. By fusion of ECFP with the granular chromogranin A, we successfully measured the pH in secretory granules of PC12 cells, and we performed a kinetic analysis of intragranular pH variations in living cells exposed to ammonium chloride.     

Nonlinear variants of the TR/BDF2 method for thermal radiative diffusion

We apply the Trapezoidal/BDF2 (TR/BDF2) temporal discretization scheme to nonlinear grey radiative diffusion. This is a scheme that is not well-known within the radiation transport community, but we show that it offers many desirable characteristics relative to other second-order schemes. Several nonlinear variants of the TR/BDF2 scheme are defined and computationally compared with the Crank–Nicholson scheme. It is found for our test problems that the most accurate TR/BDF2 schemes are those that are fully iterated to nonlinear convergence, but the most efficient TR/BDF2 scheme is one based upon a single Newton iteration. It is also shown that neglecting the contributions to the Jacobian matrix from the cross-sections, which is often done due to a lack of smooth interpolations for tabular cross-section data, has a significant impact upon efficiency.     

Pulsed erbium fiber laser with an acetylene-filled photonic crystal fiber for saturable absorption

We investigate the dynamics of an erbium-doped fiber ring laser that is equipped with an intracavity hollow core photonic crystal fiber gas cell. The cell is filled with acetylene as a saturable absorber. We observe cw operation at low pressures, Q switching at intermediate pressure levels, and mode locking at high pressures applied. Moreover, we show that the transition from the cw to the pulsed mode may be exploited for sensitive gas detection.     

Transformation of Receptor Tyrosine Kinases into Glutamate Receptors and Photoreceptors

Receptor tyrosine kinases (RTKs) are key regulators of cellular functions in metazoans. In vertebrates, RTKs are mostly activated by polypeptides but are not naturally sensitive to amino acids or light. Taking inspiration from Venus kinase receptors (VKRs), an atypical family of RTKs found in nature, we have transformed the human insulin (hIR) and hepatocyte growth factor receptor (hMET) into glutamate receptors by replacing their extracellular binding domains with the ligand‐binding domain of metabotropic glutamate receptor type 2 (mGluR2). We then imparted light sensitivity through covalent attachment of a synthetic glutamate‐based photoswitch via a self‐labelling SNAP tag. By employing a Xenopus laevis oocyte kinase activity assay, we demonstrate how these chimeric RTKs, termed light‐controlled human insulin receptor (LihIR) and light‐controlled human MET receptor (LihMET), can be used to exert optical control over the insulin or MET signaling pathways. Our results outline a potentially general strategy to convert RTKs into photoreceptors.     

Enzymatic and Molecular Characterization of Anti-Leishmania Molecules That Differently Target Leishmania and Mammalian eIF4A Proteins,LieIF4A and eIF4AMus

Previous investigations of the Leishmania infantum eIF4A-like protein (LieIF4A) as a potential drug target delivered cholestanol derivatives inhibitors. Here, we investigated the mode of action of cholesterol derivatives as a novel scaffold structure of LieIF4A inhibitors on the RNA-dependent ATPase activity of LieIF4A and its mammalian ortholog (eIF4AI). We compared their biochemical effects on RNA-dependent ATPase activities of both proteins and investigated if rocaglamide, a known inhibitor of eIF4A, could affect LieIF4A as well. Kinetic measurements were conducted at different concentrations of ATP, of the compound and in the presence of saturating whole yeast RNA concentrations. Kinetic analyses showed different ATP binding affinities for the two enzymes as well as different sensitivities to 7-α-aminocholesterol and rocaglamide. The 7-α-aminocholesterol inhibited LieIF4A with a higher binding affinity relative to cholestanol analogs. Cholesterol, another tested sterol, had no effect on the ATPase activity of LieIF4A or eIF4AI. The 7-α-aminocholesterol demonstrated an anti-Leishmania activity on L. infantum promastigotes. Additionally, docking simulations explained the importance of the double bond between C5 and C6 in 7-α-aminocholesterol and the amino group in the C7 position. In conclusion, Leishmania and mammalian eIF4A proteins appeared to interact differently with effectors, thus making LieIF4A a potential drug against leishmaniases.     

Chord length distributions in binary stochastic media in two and three dimensions

Simple models for transport through stochastic media usually assume that the chord lengths of materials are distributed exponentially. Theory predicts that, in a medium consisting of disks/spheres that can interpenetrate, chord lengths in the background material (between the disks/spheres) should exactly follow an exponential. In a medium with impenetrable (non-overlapping) disks/spheres, the distribution is only approximately exponential. This paper demonstrates, through direct numerical simulations, that for randomly distributed disks in 2D and spheres in 3D, with distributions of radii, chord lengths in the background material (between the disks/spheres) are accurately described by exponentials over five orders of magnitude when the material is dilute. The chord lengths inside the disks and spheres are not exponentially distributed, but those distributions can be calculated. A scaling relationship between the mean chord lengths in the two materials is presented for an infinite medium. By knowing the mean properties of the disks/spheres in a medium, this relationship allows one to accurately describe the statistical properties of the background material. The stochastic simulations are validated by this infinite medium relationship. When the fraction of space occupied by the disks or spheres becomes large, the distributions are no longer accurately described by an exponential.     

Recherches sur les substances mesomorphes—IX : Smectiques “fourchus”. organisation moleculaire dans les phases smectiques a

The synthesis and the mesomorphic properties of new branched smectogenic A derivatives of fluorene are described. The observation that branched chain compounds form a smectic A phase allows conclusions to be drawn on molecular arrangements in smectic A layers.     

Recherches sur les substances mesogenes—VIII : Preparation et proprietes mesomorphes de series isometriques

The synthesis and the mesomorphic properties of various series of isometric mesogenic compounds are described. It is confirmed that isometric mesogenic molecules may be nematogenic and/or smectogenic according to the position of the polar rigid core.     

Molecular modeling of the structure and dynamics of the interlayer species of ZnAlCl layered double hydroxide

Molecular dynamics simulations of the ZnAl layered double hydroxide containing interlayer chloride anions have been performed in the NpT and Np(zz)T statistical ensembles for metal Zn/Al ratios of 2 and 3. We have monitored the interlayer spacing as a function of the number of intercalated water molecules for each statistical ensemble. We have studied how these profiles are affected by the method of calculation of the charges of the hydroxide layer atoms. Diffusion coefficients of the interlayer water molecules have been calculated for different Zn/Al ratios. The calculation of the chemical potential of the interlayer water molecules has been carried out for three amounts of interlayer water molecules. The calculation showed a qualitative agreement with the bulk water chemical potential within a range of interlayer water molecule contents.