Electrochromatography in packed tubes using 1.5 to 50 μm silica gels and ODS bonded silica gels

Summary Electrochromatography (that is HPLC where the eluent is driven along the column by electro-osmosis using fields of up to 100 kV m⁻¹) promises plate efficiencies for HPLC which are comparable to those attained in capillary gas chromatography, but this requires that narrow-bore columns can be successfully packed with submicron particles. This paper demonstrates that we have now moved a considerable distance towards this goal. We show (1) that, following theory, there is no evidence of any reduction in electroosmotic velocity in columns packed with particles down to 1.5 μm diameter, (2) that reduced plate heights as low as unity are attainable for unretained solutes using both slurrypacked and drawn-packed columns 30 to 200 μm bore and up to 1 m long when packed with conventional 3 and 5 μm silica gels or with 1.5 μm impermeable silica spheres, (3) that columns driven electrically show higher plate efficiencies than identical columns driven by pressure, and (4) that 100,000 plate HPLC separations can be achieved in relatively short times of 30 minutes using in situ derivatised drawn packed capillaries containing 3 and 5 μm ODS-silica gels.     

Guanosine tetra- and pentaphosphate analysis: PEI-cellulose thin-layer purification and luciferin-luciferase liquid scintillation quantitation

A system is presented that can separate and quantitate in picomole amounts various guanosine tetra and penta phosphates namely guanosine 5′ triphosphate, 3′ diphosphate (pppGpp), guanosine 5′ tetraphosphate (ppppG), and diguanosine 5′ tetraphosphate (GppppG). It was found to be inactive with guanosine 5′ diphosphate, 3′ diphosphate (ppGpp), and a synthetic compound pCppG.The analytical detection system uses a crude firefly luciferin-luciferase system in which the various derivatives probably transphosphorylate ADP to produce the ATP necessary to emit light with the luciferin-luciferase system.The system should be useful in quantitating reactions in which guanosine tetraphosphates and pentaphosphates are involved. Their role is apparently one of control at either RNA polymerase or ribosomal levels and should be important in further research in molecular biology.     

Dipyridamole and its derivatives modify the kinetics of the electron transport in reaction centers from Rhodobacter sphaeroides

A well known vasodilator dipyridamole (DIP), 2,6-bis(diethanolamino)-4,8-dipiperidinopyrimido[5,4-d]pyrim idine, and its derivatives have recently been shown as potential co-activators (modulators) in the phenomenon of multidrug resistance (MDR) in cancer therapy. They inhibit the specific function of a transmembrane P-glycoprotein responsible for the ex-flux of anti-cancer drugs from tumor cells. To clarify molecular mechanisms of the anti-MDR activity of DIP and its two derivatives, RA25 and RA47, we have studied their effects on electron transport in reaction centers (RC) from purple photosynthetic bacteria Rb. sphaeroides, using RC as a model system. Increasing concentrations of DIP and RA47 progressively accelerate the back electron transfer from the primary quinone acceptor QA to the bacteriochlorophyll dimer Bchl2 (Bchl2+ -QA- recombination). In the absence of o-phenantroline, when both quinone acceptors QA and QB are involved in the electron transport, RA47 is more effective than DIP. DIP stabilizes the electron on the secondary quinone acceptor QB, the effect manifested as the retardation of Bchl2+ -QB- recombination. Effects of RA25 are negligible in all cases. The drugs are proposed to change the electron transport affecting the RC structural dynamics and the stabilization of the electron on quinone acceptors through modification of H-bonds in the system.     

FLUORESCENCE DECAY KINETICS and CHARACTERISTICS OF BIMOLECULAR EXCITON ANNIHILATION IN CHLOROPLASTS

Abstract— The decay profiles of the fluorescence of dark-adapted spinach chloroplasts (0^A°C) excited with single 30 ps 532 nm laser pulses of varying intensities were measured with a low-jitter streak camera system. By comparing the decay profiles of the fluorescence at low and high laser fluences, i.e. in the absence and presence, respectively, of dynamic bimolecular exciton-exciton annihilation effects, the duration of such dynamic annihilation events can be estimated. A simple model suggests that the influence of bimolecular annihilation events on the fluorescence decay kinetics should disappear within a time interval corresponding to the low intensity, unimolecular lifetime of the exciton population which is subject to exciton-exciton annihilation. The low intensity fluorescence decay profiles are characterized by three to four lifetimes (Reviewed by A. R. Holzwarth, Photochem. Photobiol. 43,707–725, 1986); it is shown here that only the shortest fluorescence components are subject to exciton annihilation, since the kinetics of the fluorescence decay are influenced by annihilations only within the initial 150–200 ps time interval after the excitation pulse. The amplitudes (but not the decay kinetics) of the longer-lived fluorescence components are decreased at high levels of laser pulse excitations, suggesting that these components are derived from the shorter-lived fluorescence decay components. The implications of these results are*discussed within the contexts of current models of the fluorescence in chloroplasts.     

Vancomycin resistance in enterococci: reprogramming of the D-ala-D-Ala ligases in bacterial peptidoglycan biosynthesis

Vancomycin binds to bacterial cell-wall intermediates to achieve its antibiotic effect. Infections of vancomycin-resistant enterococci are, however, becoming an increasing problem; the bacteria are resistant because they synthesize different cell-wall intermediates. The enzymes involved in cell-wall biosynthesis, therefore, are potential targets for combating this resistance. Recent biochemical and crystallographic results are providing mechanistic and structural details about some of these targets.     

Dark energy and the cosmic microwave background radiation

We find that current cosmic microwave background anisotropy data strongly constrain the mean spatial curvature of the Universe to be near zero, or, equivalently, the total energy density to be near critical-as predicted by inflation. This result is robust to editing of data sets, and variation of other cosmological parameters (totaling seven, including a cosmological constant). Other lines of argument indicate that the energy density of nonrelativistic matter is much less than critical. Together, these results are evidence, independent of supernovae data, for dark energy in the Universe.     

Passive harmonic mode-locked soliton fiber laser stabilized by an optically pumped saturable Bragg reflector

Stabilization of passive harmonic mode locking is achieved for what is believed to be the first time in an Er-Yb soliton fiber laser by optical pumping of the semiconductor saturable absorber above the bandgap. The results show 35-dB mode suppression of undesired harmonics.