Separations of derivatized amino acid enantiomers by cyclodextrin-modified capillary electrophoresis: mechanistic and molecular modeling studies

The enantiomers of 5-dimethylamino-1-naphthalene sulfonyl (DNS)-derivatives of selected amino acids were successfully separated using capillary electrophoresis (CE) employing cyclodextrins (CD) as enantio-selective running buffer additives. A previously described model for retention and chiral recognition in CD-modified CE is shown to adapt well in this application. Resolution of the isomers is strongly influenced by the type and concentration of cyclodextrin employed, as predicted by the model. Although data indicates differences in the electrophoretic mobilities for some of the completely complexed enantiomer pairs, selectivity generally requires exploiting differences in the amino acid-CD complexation constants for enantiomer pairs. In this work, the D-enantiomers exhibit larger formation constants and are complexed to a greater degree (elute first) at moderate CD concentration. When mixtures of amino acids are analyzed, the effects of separation conditions on general elution behavior must be considered or separated enantiomer pairs will co-elute with other enantiomers. Preliminary results aimed at predicting the strength of DNS-amino acid enantiomer-CD interactions based on molecular modeling studies are presented. A statistical mechanical approach to treating computationally derived enantiomer-CD interaction energies is shown to provide reasonable correlation with separation performance.     

Steady-state modeling of resistive-gate MOSFETs

The Pao-Sah model for the long-channel MOSFET is generalized to include MOSFETs with resistive-gate electrodes (RG-MOSFETs). The RG-MOdSFETs current-voltage characteristics are studied using this extended Pao-Sah model. The steady-state operation of the RG-MOSFET can be divided into three regimes, separated by the uniform channel condition and the pinch-off condition, respectively. The regions of operation are discussed, and a simple analytical I-V expression is given for the device     

BID: a novel BH3 domain-only death agonist

The BCL-2 family of proteins consists of both antagonists (e.g., BCL-2) and agonists (e.g., BAX) that regulate apoptosis and compete through dimerization. The BH1 and BH2 domains of BCL-2 are required to heterodimerize with BAX and to repress cell death; conversely, the BH3 domain of BAX is required to heterodimerize with BCL-2 and to promote cell death. To extend this pathway, we used interactive cloning to identify Bid, which encodes a novel death agonist that heterodimerizes with either agonists (BAX) or antagonists (BCL-2). BID possesses only the BH3 domain, lacks a carboxy-terminal signal-anchor segment, and is found in both cytosolic and membrane locations. BID counters the protective effect of BCL-2. Moreover, expression of BID, without another death stimulus, induces ICE-like proteases and apoptosis. Mutagenesis revealed that an intact BH3 domain of BID was required to bind the BH1 domain of either BCL-2 or BAX. A BH3 mutant of BID that still heterodimerized with BCL-2 failed to promote apoptosis, dissociating these activities. In contrast, the only BID BH3 mutant that retained death promoting activity interacted with BAX, but not BCL-2. This BH3-only molecule supports BH3 as a death domain and favors a model in which BID represents a death ligand for the membrane-bound receptor BAX.     

Aldosterone and dexamethasone stimulate calcineurin activity through a transcription-independent mechanism involving steroid receptor-associated heat shock proteins

1. PD 81,723 has been shown to enhance binding of adenosine to A1 receptors by stabilizing G protein-receptor coupling ('allosteric enhancement'). Evidence has been provided that in the perfused hearts and isolated atria PD 81,723 causes a sensitization to adenosine via this mechanism. 2. We have studied the effect of PD 81,723 in guinea-pig isolated atrial myocytes by use of whole-cell measurement of the muscarinic K+ current (I[K(ACh)]) activated by different Gi-coupled receptors (A1, M2, sphingolipid). PD 81,273 caused inhibition of I[K(ACh)] (IC50 approximately 5 microM) activated by either of the three receptors. Receptor-independent I[K(ACh)] in cells loaded with GTP-gamma-S and background I[K(ACh)], which contributes to the resting conductance of atrial myocytes, were equally sensitive to PD 81,723. At no combination of concentrations of adenosine and PD 81,723 could an enhancing effect be detected. 3. The compound was active from the outside only. Loading of the cells with PD 81,723 (50 microM) via the patch pipette did not affect either I[K(ACh)] or its sensitivity to adenosine. We suggest that PD 81,723 acts as an inhibitor of inward rectifying K+ channels; this is supported by the finding that ventricular I(K1), which shares a large degree of homology with the proteins (GIRK1/GIRK4) forming I[K(ACh)] but is not G protein-gated, was also blocked by this compound. 4. It is concluded that the functional effects of PD 81,723 described in the literature are not mediated by the A1 adenosine receptor-Gi-I[K(ACh)] pathway.     

The Constructed Wetland Association UK database of constructed wetland systems.

There are now more than 1,000 constructed wetland systems (CWs) in the UK. The first UK CW database was constructed by Water Research Centre (WRc) and Severn Trent Water Ltd to accompany a book on the design and performance of these systems. In that database, constructed by Gareth Job et al., only 154 beds were listed, most of which were tertiary sewage treatment sites in Severn Trent Water. The Constructed Wetland Association (CWA) was formed in 2000 as a UK water industry body in response to problems caused by unscrupulous constructors. A group of experienced, reputable designers and constructors formed the CWA to bring together best UK practice in order to counteract this problem. The group contains major water companies, designers, constructors, academics, plant growers and operators. They decided that one of the best ways of countering the problem was to assemble a database of design and performance from well-designed systems. After negotiation the CWA group took over responsibility for the database from WRc. The CWA has produced eight updates of the database which now contains information from more than 900 beds. It contains examples of the different variants of CWs in use in the UK. Most of these sites treat sewage/domestic wastewater but the database also includes examples of systems for the treatment of minewater, sludge, landfill leachate, industrial effluents, surface runoff and road runoff. Particular treatment applications are illustrated by case studies which are summary articles describing design, construction and performance.     

A vertically integrated GaAs bipolar dynamic RAM cell with storagetimes of 4.5 h at room temperature

The storage times of FET-accessed GaAs dynamic RAM cells are limited to less than 1 min at room temperature by gate leakage in the access transistor. These transistor leakage mechanisms have been eliminated by designing a vertically integrated DRAM cell in which an n-p-n bipolar access transistor is merged with a p-n-p storage capacitor. Storage times of 4.5 h are obtained at room temperature, a 1000-fold increase over the best FET-accessed cells