Spectroelectrochemical Analysis of Ion Transfer Mechanisms of Mitoxantrone at Liquid|Liquid Interfaces: Effects of Zwitterionic Dendrimer and Phospholipid Layer.

The ionic partition property and transfer mechanism of the anthraquinone antitumor agent mitoxantrone (MTX) were studied in detail at the water|1,2-dichloroethane (DCE) interface by means of surface-sensitive spectroelectrochemical techniques. The interfacial mechanism of the cationic MTX species was composed of potential-driven ion transfer and adsorption processes. The ion association between MTX and zwitterionic polyamidoamine (PAMAM) dendrimers with peripheral carboxy groups was also investigated in terms of the effects of pH and dendritic generation. The monovalent HMTX+ interacted effectively with the negatively charged dendrimers at neutral pH, while the divalent H2MTX2+ exhibited a weak association under acidic conditions. The higher stability of the dendrimer-MTX associates in the interfacial region was found for higher dendritic generations: G3.5 ≥ G2.5 > G1.5. The interfacial behavior of MTX and its dendrimer associates was further analyzed at the phospholipid-modified interface as a model biomembrane surface. The adsorption process of HMTX+ occurred mainly on the hydrophilic side of the phospholipid layer. The spectroelectrochemical results indicated that the dendrimers penetrate into the phospholipid layer and alter the transfer mechanism of HMTX+ across the interface.

A light-switching pyrene probe to detect phase-separated biomolecules.

Biomolecules may undergo liquid-liquid phase separation (LLPS) to spatiotemporally compartmentalize and regulate diverse biological processes. Because the number of tools to directly probe LLPS is limited (ie. FRAP, FRET, fluorescence microscopy, fluorescence anisotropy, circular dichroism, etc.), the physicochemical traits of phase-separated condensates remain largely elusive. Here, we introduce a light-switching dipyrene probe (Pyr-A) that forms monomers in either hydrophobic or viscous environments, and intramolecular excimers in aqueous solutions. By exploiting their distinct fluorescence emission spectra, we used fluorescent microscopic imaging to study phase-separated condensates formed by in vitro protein droplets and membraneless intracellular organelles (centrosomes). Ratiometric measurement of excimer and monomer fluorescence intensities showed that protein droplets became hydrophobic and viscous as their size increased. Moreover, centrosomes became hydrophobic and viscous during maturation. Our results show that Pyr-A is a valuable tool to characterize LLPS and enhance our understanding of phase separation underlying biological functions.

Phase-stable optical activity measurement by common-path spectral interferometry.

A robust optical activity (OA) spectrometer covering the visible and near-infrared regimes was designed and built via a combination of a linear polarizer and a birefringent plate. The OA spectrometer relies on common-path spectral interferometry, where the two interfering fields travel common optical paths, and ensures signal reproducibility over several hours. By detecting OA without polarization switching, the data acquisition time is shortened to 1 s, enabling real-time monitoring of the chiral complex formation. The present configuration also allows OA measurement with broadband pulses, which is promising for probing ultrafast circular dichroism and optical rotatory dispersion.

Aggregation-Induced Emission of Water-Soluble Tetraphenylethene Derivatives at Polarized Liquid|Liquid Interfaces.

Aggregation-induced emission (AIE) behavior of water-soluble tetraphenylethene (TPE) derivatives bearing carboxy and sulfo groups was studied at polarized liquid|liquid interfaces. The aggregation behavior of TPE derivatives in solution and at the water|1,2-dichloroethane (DCE) interface was highly dependent on their ionizable functional groups. Spectroelectrochemical analysis elucidated that the TPE derivatives were transferred across the interface accompanied by the adsorption process at the interface. The ion transfer and interfacial AIE features of TPEs responded reversibly to the externally applied potential, indicating no rigid crystalline structure formation in the interfacial region. The red shift measured in intense interfacial emission spectra demonstrated that the carboxylate derivatives formed their J-aggregates specifically at the polarized water|DCE interface, while the aggregation processes with distinguishable emission properties took place in both the interfacial region and organic solution in the sulfonate derivative system. The AIE features were also investigated at a glycerophospholipid-adsorbed interface as a model of the biomembrane surface. The aggregation process of TPE derivatives was significantly modified through the interaction with phospholipid layers which stimulate the interfacial AIE process of tetra-anionic TPEs.

Dehydration-fragmentation mechanism of cathinones and their metabolites in ESI-CID.

Various cathinone-derived designer drugs (CATs) have recently appeared on the drug market. This study examined the mechanism for the generation of dehydrated ions for CATs during electrospray ionization collision-induced dissociation (ESI-CID). The generation mechanism of dehydrated ions is dependent on the amine classification in the cathinone skeleton, which is used in the identification of CATs. The two hydrogen atoms eliminated during the dehydration of cathinone (primary amine) and methcathinone (secondary amine) were determined, and the reaction mechanism was elucidated through the deuterium labeling experiments. The hydrogen atom bonded to the amine nitrogen was eliminated with the proton added during ESI, in both of the tested compounds. This provided evidence that CATs with tertiary amine structures (such as dimethylcathinone and α-pyrrolidinophenones [α-PPs]) do not undergo dehydration. However, it was shown that the two major tertiary amine metabolites (1-OH and 2″-oxo) of CATs generate dehydrated ions in ESI-CID. The dehydration mechanisms of the metabolites of α-pyrrolidinobutiophenone (α-PBP) belongs to α-PPs were also investigated. Stable-isotope labeling showed the dehydration of the 1-OH metabolite following a simple mechanism where the hydroxy group was eliminated together with the proton added during ESI. In contrast, the dehydration mechanism of the 2″-oxo metabolite involved hydrogen atoms in three or more locations along with the carbonyl group oxygen, indicating that dehydration occurred via multiple mechanisms likely including the rearrangement reaction of hydrogen atoms. These findings presented herein indicate that the dehydrated ions in ESI-CID can be used for the structural identification of CATs.

Synergistic Ion-pair Extraction and Separation of Trivalent Lanthanoid Ions with 4-Isopropyltropolone and 1,10-Phenanthroline into o-Dichlorobenzene.

The synergistic extraction of trivalent lanthanoid (Ln(III)) ions with 4-isopropyltropolone (Hipt) and 1,10-phenanthroline (phen) in o-dichrolobenzene (DCB) was investigated. The synergistic effect in DCB is more significant than that in toluene, and the polynuclear complexes found in toluene are not formed in DCB. Based on the 3-dimensional equilibrium analysis, the extracted species for La(III), Eu(III), and Lu(III) are found to be ion-pairs, such as Ln(ipt)2phen·ClO4 and Ln(ipt)2(phen)·ClO4 in the presence of NaClO4 as a salt, and the extraction constants of the respective species were determined. The simultaneous extraction of different lanthanoids in the present extraction system was demonstrated. The separation factors between lighter lanthanoids were larger than those with 2-ethylhexyl phosphonic acid mono-2-ethylhexyl ester, an excellent extractant for the separation of lanthanoid ions, in decane.

Determination of the complex refractive index of a subwavelength-diameter platinum or gold pipe by light scattering.

The complex refractive indices of Pt and Au pipes that are subwavelength in diameter have been found to be different from those of metal thin films for the first time. The metal pipe is made from a spider silk of half-wavelength diameter clad with Pt or Au and illuminated by a plane-polarized laser of wavelength 660 nm at normal incidence. The angular distribution of the light intensity scattered by the pipe is measured and fitted using theoretical calculations based on the corresponding model. The fitting results have lead to the optimum values and uncertainty ranges of the indices and the diameter of the pipe. A field emission scanning electron microscope confirms the diameter from the optical estimation and reveals an image of the surface of the pipe.

Observation of Plasmon Wave Packet Motions via Femtosecond Time-Resolved Near-Field Imaging Techniques.

The generation and dynamics of plasmon wave packets in single gold nanorods were observed at a spatiotemporal scale of 100 nm and 10 fs via time-resolved near-field optical microscopy. Following simultaneous excitation of two plasmon modes of a nanorod with an ultrashort near-field pulse, a decay and revival feature of the time-resolved signal was obtained, which reflected the reciprocating motion of the wave packet. The time-resolved near-field images were also indicative of the wave packet motion. At some period of time after the excitation, the spatial features of the two modes appeared alternately, showing motion of plasmonic wave crests along the rod. The wave packet propagation was clearly demonstrated from this observation with the aid of a simulation model. The present experimental scheme opens the door to coherent control of plasmon-induced optical fields in a nanometer spatial scale and femtosecond temporal scale.

Local optical responses of plasmon resonances visualised by near-field optical imaging.

The unique optical characteristics of noble metal nanostructures have their origin principally in surface plasmon resonances. To exploit and design the unique characteristics arising from plasmons, an investigation of optical field structures adjacent to the nanostructure is of fundamental importance. As the spatial scale of the optical field structures is essentially smaller than the radiation wavelength in resonance with the plasmon, optical imaging methods that achieve spatial resolution beyond the diffraction limit of light are necessary to visualise the fields. In this article, we review the studies of direct experimental visualisation of plasmon resonances using near-field optical microscopy. We briefly describe the method of near-field optical microscopy used to study noble metal nanoparticles and show with several typical single gold nanoparticles that the spatial features of plasmon resonances, in particular the standing wave functions of the plasmons, can be directly visualised by near-field imaging. We then describe our recent efforts to visualise ultrafast dynamics in metal nanostructures following plasmonic excitation, which are based on near-field ultrafast imaging measurements. Another notable aspect of metal nanostructures that has attracted attention recently is the chirality of plasmons. Here, we describe a method and examples of near-field optical imaging and analyses of chiral plasmons excited on metal nanostructures.

Scattering of a light wave by a thin fiber on or near a prism: experiment and analytical theory.

We have performed an experiment of the scattering of the near field on a prism created by a laser wave, evanescent wave (EW), or plane wave (PW) of an incident angle slightly larger than or smaller than the critical angle, by a thin fiber of subwavelength diameter set above the prism, and we made an analytical theory of an adapted model for the experiment. We have been able to analyze the experimental data exactly by the model theory better than any other theory we have ever known. The importance of the multiple interaction of the wave between the fiber and the surface and also the close similarity of the scattering characteristics between the EW and the PW mentioned above have been acknowledged by the analysis of the data obtained.

Ultrafast relaxation and reaction of diiodide anion after photodissociation of triiodide in room-temperature ionic liquids.

Vibrational dephasing, vibrational relaxation, and rotational relaxation of diiodide (I(2)(-)) after photodissociation of triiodide (I(3)(-)) in room-temperature ionic liquids (RTILs) were investigated by ultrafast transient absorption spectroscopy. The vibrational energy relaxation (VER) rate of I(2)(-) produced by the photodissociation reaction of I(3)(-) was determined from the spectral profile of the transient absorption. The rates in RTILs were slightly slower than those in conventional liquids. On the other hand, the coherent vibration of I(2)(-) was not observed in RTILs, and the vibrational dephasing of the photoproduced I(2)(-) was accelerated. This was explained by the interaction between I(2)(-) and I consisting of a caged contact pair in RTILs. The orientational relaxation time of I(2)(-) determined by the transient absorption anisotropy was much longer in RTILs than in conventional liquids due to their high viscosities although the relaxation time was shorter than the prediction from the Stokes-Einstein-Debye (SED) theory. The deviation from the SED prediction was interpreted by the frequency dependence of the shear stress acting on the molecule. The dynamics of I(2)(-) in 1-butyl-3-methylimidazolium iodide ([BMIm]I) were quite different from those in other conventional RTILs: the coherent vibration of I(2)(-) was observed for the time profile of the transient absorption and the initial value of the anisotropy was reduced to 0.31 from 0.36 in conventional RTILs. These results suggest that an ultrafast reaction between the photofragment I and the solvent I(-) may occur during the photodissociation process of I(3)(-). The anomaly in the ground state coherent vibration and steady state Raman spectrum of I(3)(-) also suggest the possibility that I(3)(-) and I(-) can be located in vicinity and interact strongly with each other in [BMIm]I.

Local structures in ionic liquids probed and characterized by microscopic thermal diffusion monitored with picosecond time-resolved Raman spectroscopy.

Vibrational cooling rate of the first excited singlet (S(1)) state of trans-stilbene and bulk thermal diffusivity are measured for seven room temperature ionic liquids, C(2)mimTf(2)N, C(4)mimTf(2)N, C(4)mimPF(6), C(5)mimTf(2)N, C(6)mimTf(2)N, C(8)mimTf(2)N, and bmpyTf(2)N. Vibrational cooling rate measured with picosecond time-resolved Raman spectroscopy reflects solute-solvent and solvent-solvent energy transfer in a microscopic solvent environment. Thermal diffusivity measured with the transient grating method indicates macroscopic heat conduction capability. Vibrational cooling rate of S(1) trans-stilbene is known to have a good correlation with bulk thermal diffusivity in ordinary molecular liquids. In the seven ionic liquids studied, however, vibrational cooling rate shows no correlation with thermal diffusivity; the observed rates are similar (0.082 to 0.12 ps(-1) in the seven ionic liquids and 0.08 to 0.14 ps(-1) in molecular liquids) despite large differences in thermal diffusivity (5.4-7.5 × 10(-8) m(2) s(-1) in ionic liquids and 8.0-10 × 10(-8) m(2) s(-1) in molecular liquids). This finding is consistent with our working hypothesis that there are local structures characteristically formed in ionic liquids. Vibrational cooling rate is determined by energy transfer among solvent ions in a local structure, while macroscopic thermal diffusion is controlled by heat transfer over boundaries of local structures. By using "local" thermal diffusivity, we are able to simulate the vibrational cooling kinetics observed in ionic liquids with a model assuming thermal diffusion in continuous media. The lower limit of the size of local structure is estimated with vibrational cooling process observed with and without the excess energy. A quantitative discussion with a numerical simulation shows that the diameter of local structure is larger than 10 nm. If we combine this lower limit, 10 nm, with the upper limit, 100 nm, which is estimated from the transparency (no light scattering) of ionic liquids, an order of magnitude estimate of local structure is obtained as 10 nm < L < 100 nm, where L is the length or the diameter of the domain of local structure.

Standard optical coaxial double fiber diameter and refractive index measurement, accuracy, and precision using light scattering at normal incidence.

A plane-polarized laser wave with a wavelength of 441.6 nm illuminates a cladding optical fiber with a diameter of about 18 to 38 mum at normal incidence. A measured angular distribution of the intensity of the scattered wave corresponds well with the differential cross section of a rigorous theoretical calculation of a coaxial double cylinder over a wide range of scattering angle. The diameter and refractive index of the cladding and core of the illuminated part of a fiber have been determined accurately for each uncertainty.

Charge effect on the diffusion coefficient and the bimolecular reaction rate of diiodide anion radical in room temperature ionic liquids.

The diffusion coefficients of diiodide anion radical, I(2)(-), in room temperature ionic liquids (RTILs) were determined by the transient grating (TG) method using the photochemical reaction of iodide. The diffusion coefficients we obtained were larger in RTILs than the theoretical predictions by the Stokes-Einstein relation, whereas both values are similar in conventional solvents. By comparison with the diffusion coefficients of neutral molecules, it was suggested that the Coulomb interaction between I(2)(-) and constituent ions of RTILs strongly affects the diffusion coefficients. The bimolecular reaction rates between I(2)(-) were calculated by the Debye-Smoluchowski equation using the experimentally determined diffusion coefficients. These calculated reaction rate were much smaller than the experimentally determined rates (Takahashi, K.; et al. J. Phys. Chem. B 2007, 111, 4807), indicating the charge screening effect of RTILs.

Pressure-induced changes in the structure and function of the kinesin-microtubule complex.

Kinesin-1 is an ATP-driven molecular motor that "walks" along a microtubule by working two heads in a "hand-over-hand" fashion. The stepping motion is well-coordinated by intermolecular interactions between the kinesin head and microtubule, and is sensitively changed by applied forces. We demonstrate that hydrostatic pressure works as an inhibitory action on kinesin motility. We developed a high-pressure microscope that enables the application of hydrostatic pressures of up to 200 MPa (2000 bar). Under high-pressure conditions, taxol-stabilized microtubules were shortened from both ends at the same speed. The sliding velocity of kinesin motors was reversibly changed by pressure, and reached half-maximal value at approximately 100 MPa. The pressure-velocity relationship was very close to the force-velocity relationship of single kinesin molecules, suggesting a similar inhibitory mechanism on kinesin motility. Further analysis showed that the pressure mainly affects the stepping motion, but not the ATP binding reaction. The application of pressure is thought to enhance the structural fluctuation and/or association of water molecules with the exposed regions of the kinesin head and microtubule. These pressure-induced effects could prevent kinesin motors from completing the stepping motion.

Light scattering from a birefringent cylinder, spider silk, slimmer than the wavelength approaches dipole radiation.

Using a plane-polarized laser of wavelength lambda = 543.5 or 441.6 nm and spider silk, we investigated the diffraction of a transparent cylinder of diameter D approximately lambda - lambda/5 at normal incidence. The measured pattern corresponded well to the one calculated by a rigorous solution of the theory for the problem. The birefringent index and D of the sample could be determined simultaneously. The experimental data of the scattering cross section for D < lambda/4 suggested that the data approached dipole radiation.