S-homoadenosyl-L-cysteine and S-homoadenosyl-L-homocysteine. Synthesis and binding studies of hon-hydrolyzed substrate analogues with S-adenosyl-L-homocysteine hydrolase

Treatment of homoadenosine [9-(5-deoxy-beta-D-ribo-hexofuranosyl)adenine] with thionyl chloride and pyridine in acetonitrile gave 6'-chloro-6'-deoxyhomoadenosine, which underwent nucleophilic displacement with L-cysteine or L-homocysteine to give homologated analogues of S-adenosyl-L-homocysteine. Each amino acid in aqueous sodium hydroxide at 60 degrees C gave excellent conversion from the chloronucleoside, and adsorption on Amberlite XAD-4 resin provided more convenient isolation than prior methods. Weak binding of these non-hydrolyzed analogues to S-adenosyl-L-homocysteine hydrolase was observed.     

Mixed finite element methods for computing groundwater velocities

Central to the understanding of problems in water quality and quantity for effective management of water resources is the development of accurate numerical models to stimulate groundwater flows and contaminant transfer. We discuss several important difficulties arising in modeling of subsurface flow and present promising numerical procedures for alleviating these problems. Furthermore, we describe mixed-finite element techniques for accurately approximating fluid velocities, and review computational results on a variety of hydrologic problems.     

Vibration-induced mobilization of trapped oil ganglia in porous media: experimental validation of a capillary-physics mechanism

The development of methods for mobilizing residual organic liquids trapped in porous media is becoming increasingly important as world demand for oil increases and because of the need to remediate aquifers degraded by slow-dissolving organic contaminants. Low-frequency elastic wave stimulation is one such technique, but until recently the lack of a mechanistic understanding of the effects of vibration on mobilization of oil ganglia has prevented the method from being applied predictably in the field. Recently, a simple capillary-physics mechanism has been developed to explain vibration-induced mobilization of a trapped non-wetting organic phase in porous media. Specific predictions that follow from this hypothesized mechanism are that vibrations will be most effective in mobilizing trapped oil when the acceleration amplitude is within an optimal range of values (that depend on the magnitudes of the capillary forces trapping the ganglia and the imposed static pressure gradients) and for sufficiently low vibration frequencies. In this paper we describe two-dimensional glass micromodel experiments that support these predictions.     

Simultaneous detection of vesicular content and exocytotic release with two electrodes in and at a single cell

We developed a technique employing two electrodes to simultaneously and dynamically monitor vesicular neurotransmitter storage and vesicular transmitter release in and at the same cell. To do this, two electrochemical techniques, single-cell amperometry (SCA) and intracellular vesicle impact electrochemical cytometry (IVIEC), were applied using two nanotip electrodes. With one electrode being placed on top of a cell measuring exocytotic release and the other electrode being inserted into the cytoplasm measuring vesicular transmitter storage, upon chemical stimulation, exocytosis is triggered and the amount of release and storage can be quantified simultaneously and compared. By using this technique, we made direct comparison between exocytotic release and vesicular storage, and investigated the dynamic changes of vesicular transmitter content before, during, and after chemical stimulation of PC12 cells, a neuroendocrine cell line. While confirming that exocytosis is partial, we suggest that chemical stimulation either induces a replenishment of the releasable pool with a subpool of vesicles having higher amount of transmitter storage, or triggers the vesicles within the same subpool to load more transiently at approximately 10–20 s. Thus, a time scale for vesicle reloading is determined. The effect of l-3,4-dihydroxyphenylalanine (l-DOPA), the precursor to dopamine, on the dynamic alteration of vesicular storage upon chemical stimulation for exocytosis was also studied. We found that l-DOPA incubation reduces the observed changes of vesicular storage in regular PC12 cells, which might be due to an increased capacity of vesicular transmitter loading caused by l-DOPA. Our data provide another mechanism for plasticity after stimulation via quantitative and dynamic changes in the exocytotic machinery.

Simultaneous measurements of IVIEC and SCA by two nanotip electrodes allows direct and dynamic comparison between vesicular transmitter content and vesicular transmitter release to shed light on stimulation-induced plasticity.