Measurement of intracellular Ca2+ changes using novel caged cyclic nucleotides and confocal laser scanning microscopy.

The intention of this study is to explore the applicability of confocal microscopy in conjunction with the use of caged cyclic nucleotide derivatives. The methodological potential of UV laser confocal microscopy has been assessed. It is shown that illumination of a single cell or a small area of a single cell is possible, whereby the intracelluar Ca2+ signal is measured at illuminated and non-illuminated cells. Such measurements do not have a high time resolution because of the specific system parameters. However, with an N2 pulse laser (not part of the standard microscope set-up), Ca2+ signals with a time resolution of around 100 ms have been measured. This facilitates investigation of the kinetics of Ca2+ influx. Intracellular Ca2+ measurements at HEK293 and sperm cells have been made here. For sperm cells the advantages of confocal microscopy are best evidenced in conjunction with the use of caged cyclic nucleotides; a cyclic nucleotide-gated Ca2+ influx at the tail of these cells has thereby been demonstrated for the first time.     

Microphase Separation within a Comb Copolymer with Attractive Side Chains: A Computer Simulation Study

Computer simulation modelling of a flexible comb copolymer with attractive interactions between the monomer units of the side chains is performed. The conditions for the coil‐globule transition, induced by the increase of attractive interaction, ε, between side chain monomer units, are analysed for different values of the number of monomer units in the backbone, N, in the side chains, n, and between successive grafting points, m. It is shown that the coil‐globule transition of such a copolymer corresponds to a first‐order phase transition. The energy of attraction (ε) required for the realisation of the coil‐globule transition decreases with increasing n and decreasing m. The coil‐globule transition is accompanied by significant aggregation of side chain units. The resulting globule has a complex structure. In the case of a relatively short backbone (small value of N), the globule consists of a spherical core formed by side chains and an enveloping shell formed by the monomer units of the backbone. In the case of long copolymers (large value of N), the side chains form several spherical micelles while the backbone is wrapped on the surfaces of these micelles and between them.     

Microstructural studies on degradation of interface between LSM–YSZ cathode and YSZ electrolyte in SOFCs

The changes in the cathode/electrolyte interface microstructure have been studied on anode-supported technological solid oxide fuel cells (SOFCs) that were subjected to long-term (1500 h) testing at 750 °C under high electrical loading (a current density of 0.75 A/cm²). These cells exhibit different cathode degradation rates depending on, among others, the composition of the cathode gas, being significantly smaller in oxygen than in air. FE-SEM and high resolution analytical TEM were applied for characterization of the interface on a submicron- and nano-scale. The interface degradation has been identified as the loss of LSM coverage and the loss of three-phase-boundary (TPB) length. Firstly, the degradation is caused by the size reduction of the individual LSM/YSZ electrolyte contact points (areas) that are initially of 100–200 nm in diameter. Quantitative microstructure evaluation shows that in the cell tested in air this mechanism contributes to an estimated overall reduction in the LSM coverage and the TPB length by 50 and 30%, respectively. For the cell tested in oxygen the corresponding values are 10 and 4%. Secondly, in the cell tested in air the LSM coverage and the TPB length appear to decrease further due to the more pronounced formation of insulating zirconate phases that are present locally and preferably in LSM/YSZ electrolyte contact areas. The effects of the cathode gas on the interface degradation are discussed considering the change of oxygen activity at the interface, possible changes in the Mn diffusion pattern as well as the LSM/YSZ reactivity. Finally, based on thermodynamic calculations a T–p(O₂) diagram predicting the safe and risky operation conditions in terms of the zirconate formation is presented and compared with the experimental observations.     

Increasing the pump-up rate to polarize He gas using spin-exchange optical pumping method

In recent years, polarized ³He gas has increasingly been used as neutron polarizers and polarization analyzers. Two of the leading methods to polarize the ³He gas are the spin-exchange optical pumping (SEOP) method and the meta-stable exchange optical pumping (MEOP) method. At present, the SEOP setup is comparatively compact due to the fact that it does not require the sophisticated compressor system used in the MEOP method. The temperature and the laser power available determine the speed, at which the SEOP method polarizes the ³He gas. For the quantity of gas typically used in neutron scattering work, this speed is independent of the quantity of the gas required, whereas the polarizing time using the MEOP method is proportional to the quantity of gas required. Currently, using the SEOP method to polarize several bar-liters of ³He to 70% polarization would require 20−40 h. This is an order of magnitude longer than the MEOP method for the same quantity of gas and polarization. It would therefore be advantageous to speed up the SEOP process. In this article, we analyze the requirements for temperature, laser power, and the type of alkali used in order to shorten the time required to polarize ³He gas using the SEOP method.     

Electrochemical reduction of NO on La2-x Sr x NiO4 based electrodes

The series La_2 − x Sr _x NiO₄ (x = 0.0, 0.05, 0.15, 0.25, 0.35, and 1.0) was tested for functionality as electrode materials for direct electrochemical reduction of NO. The materials were tested using cyclic voltammetry in 1% NO and 10% O₂ in Ar on a cone-shaped electrode. The best materials for the electrochemical reduction of NO are La₂NiO₄ and LaSrNiO₄, which have current densities for NO reduction 1.82 and 7.09 times higher, respectively, than for O₂ at 400 °C. Increasing the temperature decreased the ability to reduce NO before O₂ while the activity increased. The adsorbed species during direct decomposition was attempted, clarified using X-ray absorption near-edge structure experiments and thermogravimetry, but no conclusive results were obtained.     

Microphase separation in multiblock copolymer melts: Nonconventional morphologies and two-length-scale switching

The phase behavior of AfmN(BN2AN2)B(1-fmN) multiblock copolymer melts is studied within the weak segregation theory. The interplay between ordering on different length scales is shown to cause dramatic changes both in the ordered phase symmetry and periodicity upon small variation of the architectural parameters of the macromolecules. Phase diagrams are presented in the (f,chiN) plane (chi is the Flory-Huggins parameter) for various values of the architecture parameters n and m. Near the critical surface, i.e., for (f-0.5)2<1, such nonconventional cubic phases as the face-centered cubic (FCC), simple cubic (SC), (double) gyroid, and the so-called BCC(2) (single gyroid) are found to be stable. The lamellar morphology is shown to be replaced by BCC2, FCC, or SC (depending on the structural parameters) as the most stable low-temperature phase.     

Spectroscopy and electrochemistry of cytochrome P450 BM3-surfactant film assemblies

We report analyses of electrochemical and spectroscopic measurements on cytochrome P450 BM3 (BM3) in didodecyldimethylammonium bromide (DDAB) surfactant films. Electronic absorption spectra of BM3-DDAB films on silica slides reveal the characteristic low-spin FeIII heme absorption maximum at 418 nm. A prominent peak in the absorption spectrum of BM3 FeII-CO in a DDAB dispersion is at 448 nm; in spectra of aged samples, a shoulder at approximately 420 nm is present. Infrared absorption spectra of the BM3 FeII-CO complex in DDAB dispersions feature a time-dependent shift of the carbonyl stretching frequency from 1950 to 2080 cm(-1). Voltammetry of BM3-DDAB films on graphite electrodes gave the following results: FeIII/II E(1/2) at -260 mV (vs SCE), approximately 300 mV positive of the value measured in solution; DeltaS degrees (rc), DeltaS degrees , and DeltaH degrees values for water-ligated BM3 in DDAB are -98 J mol(-1) K(-1), -163 J mol(-1) K(-1), and -47 kJ mol(-1), respectively; values for the imidazole-ligated enzyme are -8 J mol(-1) K(-1), -73 J mol(-1) K(-1), and -21 kJ mol(-1). Taken together, the data suggest that BM3 adopts a compact conformation within DDAB that in turn strengthens hydrogen bonding interactions with the heme axial cysteine, producing a P420-like species with decreased electron density around the metal center.