Computerized Drug Screening of the S31N Mutant of the Influenza A Virus

Influenza is responsible for millions of hospitalizations and deaths every year. Treating the influenza virus has become increasingly difficult since mutant viruses have rendered previously viable drugs ineffective. New drug-screening procedures are now necessary in order to speed up the process of viable drug discovery. Molecular simulations have proven valuable for studying drug effectiveness in wild-type flu viruses, but little has been done on prevailing influenza mutants. Influenza mutants often feature a change in the M2 proton channel, a critical component in the viral replication and infection cycle. To counter this, drugs must be discovered that can block the influenza M2 channel of mutant viruses. The purpose of this research report is to discuss the methods and results of using molecular dynamics simulations to predict drug efficacy against the M2 proton channel of the Influenza A virus

Sodium Flux Ratio in Voltage-Clamped Squid Giant Axons

The sodium flux ratio across the axolemma of internally perfused, voltage-clamped giant axons of Loligo pealei has been measured at various membrane potentials. The flux ratio exponent obtained from these measurements was about unity and independent of membrane voltage over the 50 mV range from about -20 to +30 mV. These results, combined with previous measurements of ion permeation through sodium channels, show that the sodium channel behaves like a multi-ion pore with two ion binding sites that are rarely simultaneously occupied by sodium

Two-, Three-, and Four-Particle Spatial Correlations Among Tertiary Cosmic Ray Muons

Showers of muons which result from interactions of primary cosmic rays high in the atmosphere appear deep underground as bundles of nearly parallel muons. For threshold energies of 1 TeV and arrival zenith angles of 45 degrees, those events having a small mean number of muons are described by a power law distribution in shower size having an exponent of ≅ -3.7 and a radial density distribution that decreases with distance R from the shower axis a little less rapidly than exp(-R/5.4 m). Comparison of new data on four muon spatial correlations in terms of the same phenomenology with data on two and three muon spatial correlations from previous experiments suggests that the shower size distribution steepens and the shower radial density distribution broadens with increasing shower size (i.e., with increasing mean energy of the primary cosmic ray at a given threshold energy of the muons observed). These results might be explained by a basically geometrical effect due either to a total inelastic cross section which rises with energy or by an enhanced contribution from the interactions of secondaries. If the effect does not have a geometrical origin, then it may be necessary to reformulate the input to shower development calculations which have assumed that interaction products axe described by factored, uncorrelated single particle statistical distributions. The relationships between the inclusive distributions predicted by theory and those of the empirical phenomenology are explored in.some detail

Fluorescence anisotropy of diphenylhexatriene and its cationic Trimethylamino derivative in liquid dipalmitoylphosphatidylcholine liposomes: opposing responses to isoflurane

<p>Abstract</p> <p>Background</p> <p>The mechanism of action of volatile general anesthetics has not yet been resolved. In order to identify the effects of isoflurane on the membrane, we measured the steady-state anisotropy of two fluorescent probes that reside at different depths. Incorporation of anesthetic was confirmed by shifting of the main phase transition temperature.</p> <p>Results</p> <p>In liquid crystalline dipalmitoylphosphatidylcholine liposomes, isoflurane (7-25 mM in the bath) increases trimethylammonium-diphenylhexatriene fluorescence anisotropy by ~0.02 units and decreases diphenylhexatriene anisotropy by the same amount.</p> <p>Conclusions</p> <p>The anisotropy data suggest that isoflurane decreases non-axial dye mobility in the headgroup region, while increasing it in the tail region. We propose that these results reflect changes in the lateral pressure profile of the membrane.</p

Copper Complexes as Influenza Antivirals: Reduced Zebrafish Toxicity

Copper complexes have previously been developed to target His37 in influenza M2 and are effective blockers of both the wild type (WT) and the amantadine-resistant M2S31N. Here, we report that the complexes were much less toxic to zebrafish than CuCl2. In addition, we characterized albumin binding, mutagenicity, and virus resistance formation of these metal complexes, and employed steered molecular dynamics simulations to explore whether the complexes would fit in M2. We also examined their anti-viral efficacy in a multi-generation cell culture assay to extend the previous work with an initial-infection assay, discovering that this is complicated by cell culture medium components. The number of copper ions binding to bovine serum albumin (BSA) correlates well with the number of surface histidines and BSA binding affinity is low compared to M2. No mutagenicity of the complexes was observed when compared to sodium azide. After 10 passages of virus in MDCK culture, the EC50 was unchanged for each of the complexes, i.e. resistance did not develop. The simulations revealed that the compounds fit well in the M2 channel, much like amantadine

Advances in the operation of the DIII-D neutral beam computer systems

The DIII-D neutral beam system routinely provides up to 20 MW of deuterium neutral beam heating in support of experiments on the DIII-D tokamak, and is a critical part of the DIII-D physics experimental program. The four computer systems previously used to control neutral beam operation and data acquisition were designed and implemented in the late 1970`s and used on DIII and DIII-D from 1981--1996. By comparison to modern standards, they had become expensive to maintain, slow and cumbersome, making it difficult to implement improvements. Most critical of all, they were not networked computers. During the 1997 experimental campaign, these systems were replaced with new Unix compliant hardware and, for the most part, commercially available software. This paper describes operational experience with the new neutral beam computer systems, and new advances made possible by using features not previously available. These include retention and access to historical data, an asynchronously fired ``rules`` base, and a relatively straightforward programming interface. Methods and principles for extending the availability of data beyond the scope of the operator consoles will be discussed

Permeation characteristics of gramicidin conformers

To investigate the molecular origin of decreased conductance in variant gramicidin channels, we examined the current-voltage (IV) characteristics of single Val1-gramicidin A channels. Unlike standard channels, all variant channels showed pronounced rectification even though bathing solutions were symmetrical. Moreover, channels of lower conductance consistently showed more pronounced rectification. Analysis within the framework of a three-barrier, two-site, single-filing model indicates that the shape of the variant channel IVs could be best explained by an increase in binding affinity near one of the two channel entrances. This conclusion was further tested by characterizing single channel IVs in bi-ionic solutions having different cationic species at each channel entrance. In Cs/Na bi-ionic solutions, reversal potentials of variant channels often differed by a small but significant amount from those of standard channels. When a membrane potential was applied, the ionic currents tended to be reduced more when flowing from the Na+ side than the Cs+ side. These observations support the conclusion that variant channels have increased binding affinity at one end of the channel. Furthermore, H+ currents were increased while Ag+ currents were unaltered for most variant channels exhibiting decreased Na+ or Cs+ currents. The increased H+ conductance argues against long-range coulombic forces as the basis for decreased Na+ or Cs+ conductance while the normal Ag+ conductance suggests that the binding site field strength increases by a change in carbonyl geometry at the channel entrance