Subdiffraction imaging through the selective donut-mode depletion of thermally stable photoswitchable fluorophores: numerical analysis and application to the fluorescent protein Dronpa

The fast and reversible on/off switching of the fluorescence emission of the GFP-like fluorescent protein Dronpa has attracted considerable interest for applications in subdiffraction imaging. In this paper we study the use of a donut-mode beam in combination with two more overlapping laser beams to increase the imaging resolution through selective switching to the nonfluorescent photoswitched state. We devise and run a series of numerical simulations to determine suitable photophysical parameters of prospective, thermally stable photoswitchable molecules, in terms of photoswitching quantum yields, fatigue resistance, and possible presence of transient nonfluorescent states. Many of our findings are applicable to other measurements that make use of donut beams, and these guidelines can be used in the synthesis and screening of novel photoswitchable compounds. We experimentally demonstrate the possibility of obtaining increased resolution by making use of the efficient and thermally stable Dronpa photoswitching, using equipment that is commonly available.     

Conformation and location of membrane-bound salinomycin-sodium complex deduced from NMR in isotropic bicelles

An ionophore antibiotic salinomycin was studied in a membrane environment consisting of isotropic bicelles, a better model for biological membranes than micelles, and its conformation and topological orientation in bicelles was determined. 2D NMR measurements and restrained conformational search revealed that salinomycin-sodium salt in bicelles adopts an open conformation in which the orientation of the E-ring is significantly different from that in crystal and solution structures. This conformational alteration breaks an intramolecular hydrogen bond between 28-OH and 1-O, dislocates the ether oxygen of the E-ring from a coordinated position to the sodium ion observed in the crystal, and consequently weakens the complexation between salinomycin and the sodium ion. Paramagnetic relaxation experiments using doxyl-phospholipids reveal that salinomycin is embedded shallowly in bicelles, with both terminals being closer to the water interface and the olefin portion facing the bicelle interior. Measurements of intermolecular NOEs between salinomycin and phospholipids further supported this orientation. Weaker complexation with sodium ion and positional preference in the membrane polar region may facilitate the catch-and-release of metal ions at the polar/nonpolar interface of bilayers. On the basis of these findings, a model for salinomycin-assisted transport of metal ions across biological membranes is proposed.     

A hetero-core structured fiber optic pH sensor

Novel spectroscopic sensor based on a hetero-core structured fiber optic is described in this paper. The hetero-core structured fiber optic consists of multi mode fibers and a short piece of single mode fiber which was inserted in the multi mode fibers. Phenol red and/or cresol red as pH sensitive dyes were immobilized on the surface of the hetero-core portion by using sol-gel method, and the pH change detection was performed by immersing the hetero-core portion into the solution. In the case that the cresol-red immobilized fiber was immersed in the alkaline and/or acidic solution, the peak wavelength of the propagating loss spectra were about 575 and 545 nm, respectively. These propagating loss spectra were similar to that of the absorbance spectra of the dye solution. In the propagating loss spectra of phenol-red immobilized fiber, these spectra were similar to that of the dye solution. The colorimetric change of the dye in the support matrix was reversible, and the response time of the sensor was within 30 s.     

Structural development of liver X receptor (LXR) antagonists derived from thalidomide-related glucosidase inhibitors

Following our previous discovery of LXR antagonistic activity of 2'-substituted phenylphthalimides derived from thalidomide-related glucosidase inhibitors, structure-activity studies and further structural development led to 5-chloro-N-2'-n-pentylphenyl-1,3-dithiophthalimide (5CPPSS-50), with IC50 values of about 10 and 13 microM for LXRalpha and LXRbeta, respectively.     

Adsorption behaviors of high-valence metal ions on desferrioxamine B immobilization nylon 6,6 chelate fiber under highly acidic conditions

Adsorption behaviors of the high-valence metal ions Zr(IV), Hf(IV), Ti(IV), V(V), Nb(V), Ta(V), and Mo(IV) on desferrioxamine B (DFB) immobilization nylon 6,6 chelate fiber was investigated under highly acidic conditions. Though the complexes of DFB and the high-valence metal ions were extracted without selectivity by solvent extraction, the only zirconium ions showed higher adsorption percentages than that of other high-valence metal ions on the DFB immobilization nylon 6,6 chelate fiber. Adsorption properties were caused that limited the freedom of DFB by chemical immobilization. Especially, hafnium ions and zirconium ions, which have similar chemical properties, showed different adsorption behavior in highly acidic aqueous solutions. Zirconium ions were quantitatively adsorbed up to 13.5 micromol/g.     

Observation of characteristic IR band assignable to dimerized copper ions in montmorillonite

Copper ion-exchanged montmorillonite clays, which had been prepared by ion exchange in an aqueous solution of CuCl(2) at a temperature above 323 K, exhibited the characteristic IR band in the region 3360-3310 cm(-1). No such bands were observed for the samples prepared by using different ion-exchange solutions and at different temperatures. From the spectroscopic observations, it was revealed that the ion-exchanged copper ions (Cu(2+)) are in the form of dimerized species by bridging two hydroxyls, [Cu(2+) <(OH(-)(2) > Cu(2+)], in montmorillonite.     

Denaturation of alpha-lactalbumin and ubiquitin studied by electrospray and laser spray

Electrospray and laser spray mass spectra of human alpha-lactalbumin and bovine ubiquitin were studied, with an emphasis on the denaturation induced by laser spray. There were no remarkable differences in the electrospray and laser spray mass spectra for acidic and basic aqueous solutions of alpha-lactalbumin in positive and negative modes of operations. This originates from the fact that this protein is tightly folded with four disulfide bonds. For ubiquitin, however, denaturation was induced by laser spray for the positive mode of operation and the [M+nH](n+) with a maximum of n = 13 was observed, i.e., all the acidic amino acid residues are fully neutralized (protonated). In contrast, the laser-induced denaturation was not observed for the negative mode of operation, i.e., denaturation of ubiquitin is largely suppressed in the negatively charged liquid droplets. The marked difference observed in the positive and negative modes of operations for ubiquitin is ascribed to the difference in the susceptibility of side-chain/main-chain interactions in the positive-ion excess and in the negative-ion excess liquid droplets. That is, the interactions between the basic residues and main-chain amide carbonyl groups (-NH(3) (+)***O=C< or -NH(2)***O=C<) which play an important role in stabilizing the protein structures are not so affected in the negative mode of operation but are weakened in the positive mode of operation.     

Catalytic <Emphasis Type="Italic">N</Emphasis>-Alkylation of Amines with Primary Alcohols over Halide Clusters

Piperidine was reacted with methanol under a hydrogen stream in the presence of (H₃O)₂[(W₆Cl₈)Cl₆]·6H₂O supported on silica gel. When the temperature was raised above 200 °C, the catalytic activity of the cluster appeared. Piperidine N-methylation proceeded yielding N-methylpiperidine in 95% selectivity at 350 °C. The corresponding halide clusters of niobium, molybdenum, and tantalum supported on silica gel also catalyzed the reaction. Primary alcohols such as ethanol and 1-propanol produced the corresponding N-alkyl products of piperidine; however, secondary and tertiary alcohols did not. Selective N-methylation of pyrrolidine, hexamethyleneimine, butylamine, and aniline also proceeded. Thus, the clusters catalyzed alkylation of aliphatic, alicyclic, and aromatic amines with primary alcohols. A Brønsted acid site attributable to a hydroxo ligand, which is formed on the cluster complex by thermal activation, is proposed as the active site of the catalyst.