A study into the effects of protein binding on nucleotide conformation

In this study, we examine the effects of binding to protein upon nucleotide conformation, by the comparison of X-ray crystal structures of free and protein-bound nucleotides. A dataset of structurally non-homologous protein-nucleotide complexes was derived from the Brookhaven Protein Data Bank by a novel protocol of dual sequential and structural alignments, and a dataset of native nucleotide structures was obtained from the Cambridge Structural Database. The nucleotide torsion angles and sugar puckers, which describe nucleotide conformation, were analysed in both datasets and compared. Differences between them are described and discussed. Overall, the nucleotides were found to bind in low energy conformations, not significantly different from their 'free' conformations except that they adopted an extended conformation in preference to the 'closed' structure predominantly observed by free nucleotide. The archetypal conformation of a protein-bound nucleotide is derived from these observations.     

The novel inotropic agent CGP 48506 alters force primarily by Ca(2+)-independent mechanisms in porcine skinned trabeculae

OBJECTIVES: The aim was to determine whether, and by what mechanism(s), a novel inotropic agent 5-methyl-6-phenyl-1,3,5,6-tetrahydro-3, 6-methano-1,5-benzodiazocine-2,4-dione (BA 41899) and its enantiomers directly alter the Ca2+ sensitivity of cardiac myofilaments. METHODS: Porcine ventricular trabeculae were permeabilised with Triton X-100. The relationship between force and pCa (-log[Ca2+]) was determined in the presence and absence of ATP. Troponin I was extracted, using vanadate, to produce unregulated maximally activated myofilaments. Force and actomyosin ATPase activity were measured simultaneously to determine tension cost (ATPase activity/tension). The effects of the (+) enantiomer (CGP 48506) on the twitch of intact muscle were demonstrated using rat papillary muscle. RESULTS: 100 microM BA 41899 had a pronounced Ca2+ sensitising effect on force production by porcine skinned cardiac fibres, increasing the pCa required for 50% maximal activation by 0.64 units, while suppressing maximum force by 18.3%. Resting tension was unaffected. These actions were primarily caused by CGP 48506 and were concentration dependent. At concentrations less than 100 microM, CGP 48506 also increased twitch amplitude in intact papillary muscles with no effect on resting tension, whereas 100 microM CGP 48506 increased resting force due to a slowing of relaxation. 100 microM CGP 48506 potentiated Ca(2+)-independent rigor tension in skinned trabeculae, indicating a Ca2+ sensitising mechanism unrelated to Ca2+ binding to troponin C. Tension cost was unaffected by 100 microM CGP 48506 over the entire range of activating Ca2+ concentrations. Suppression of maximum force by CGP 48506 was independent of both Ca2+ concentration and the regulatory troponin complex. CONCLUSIONS: Both the increase in Ca2+ sensitivity during submaximal activation and the depression of maximum force which are induced by CGP 48506 in skinned trabeculae occur at least partly through Ca(2+)-independent mechanisms.     

A new minimum zone method for evaluating flatness errors

A new minimum zone method for flatnes error analyis is proposed in this article. Based on the criteria for the minimum zone solution and strict rules for data exchange, a simple and rapid algorithm, called the control plane rotation scheme, is developed for the flatness analysis of a flat surface. Experimental work was performed, and some examples are given in terms of the minimum zone and least-squares solutions.     

Combustion synthesis of advanced materials: Part I. Reaction parameters

An explanation of combustion (self propagating high temperature) synthesis (SHS) is given together with a historical perspective of the examination of such exothermic reactions. The application of thermochemical functions has been used to predict theoretically the maximum adiabatic temperature, T_ad. This, combined with a knowledge of the ignition temperature, T_ig, and the actual combustion temperature, T_c, has been used to determine the heat loss from the SHS reaction and the amount of heat needed to raise the adjacent, cold, reactant layer to the ignition temperature in order to maintain the self sustaining nature of the propagating mode of the reaction. The pertinent reaction parameters that control self propagating high temperature (combustion) synthesis reactions have been examined. These include: reactant particle size and shape; powder mixing and compaction; green density; reaction stoichiometry; impurities; volatiles and diluents; reaction environment; mode and technique of ignition; heating rate; and the effect of these parameters on the generation of heat, exothermicity and control of the SHS reaction.     

Analysis of the effect of shrinkage on macrosegregation in alloy solidification

Numerical calculations based on a continuum model are used to examine the effects of solidification shrinkage on the redistribution of solute in a Pb-19.2 pct Sn mixture which is convectively cooled at a sidewall. For each of three different cooling rates, separate calculations are performed for shrinkage and buoyancy-induced flows, as well as for the combined influence of shrinkage and buoyancy effects. The calculations reveal that flow and macrosegregation patterns are more strongly influenced by buoyancy effects over a wide range of solidification rates. Although extremely large solidification rates yield small regions near the chilled wall in which shrinkage-induced flows control the redis-tribution of solute, the overall effect on macrosegregation is small relative to that associated with buoyancy. Scaling analysis of the governing equations produces reference shrinkage and buoyancy velocities which can be used to extend the current numerical results to other binary systems.     

Comparisons of taste perceptions and preferences of Japanese and Australian consumers: overview and implications for cross-cultural sensory research

A program of research that compared the taste perceptions and preferences of Japanese and Australian consumer panels is reviewed from the point of view of the general issues that have emerged. These studies revealed few cross-cultural differences in the perceptions of the panels, implicating an important role for dietary experience in differences in preference. There was also little evidence for cross-cultural influences on panellists' assessment behaviour, such as scale usage. Studies of predominantly sweet and salty foods from both Australia and Japan illustrated the importance of familiarity with the overall product as an influence on the assessment of individual sensory characteristics. Subsequent attempts to overcome this influence by comparing cross-cultural responses to the manipulation of tastes within foods common to both cultures revealed no differences in the optimum tastant level within each of the foods. The implications of these studies for future cross-cultural research is addressed, and it is argued that future studies need to develop methods for determining the acceptability of relatively novel foods specifically developed for export markets, and also to address the complexity of food preferences from the point of view of both sensory and non-sensory influences.     

Lysis of human monocytic leukemia cells by extracellular adenosine triphosphate: mechanism and characterization of the adenosine triphosphate receptor

The present study shows that extracellular adenosine triphosphate (ATP) has the capacity to mediate dose-dependent lysis of the monocytic leukemia cell line THP-1. The lysis, assessed by 51Cr release, was found to be selective for ATP, because adenosine diphosphate (ADP) or other nucleotides were less effective in their ability to lyse the cells. The amount of 51Cr released was particularly enhanced by the stimulation of the cells with 1,000 U/mL of interferon gamma (IFN-gamma) for 3 days, and the sensitivity was time and dose dependent. Analysis of the mechanism of lysis indicated that the fully ionized form, ATP4-, mediated the lysis, because the addition of cation chelators or the absence of the divalent cations, Ca2+ and Mg2+, in the culture medium of a 6-hour 51Cr release assay increased the percent specific lysis. Therefore, the ATP receptors on THP-1 cells were classified as P2Z purinoceptors. Moreover, it is shown here that the Ca2+/calmodulin complex plays a role in the regulation of the lysis by extracellular ATP of THP-1 cells, because antagonists of this complex, such as trifluoperazine or KN-62, were found to inhibit the ATP-mediated cell lysis.     

Response spectrum analysis of piping systems with elastic-plastic gap supports

A new seismic support device and its application in piping systems is described. The device, E-BAR (patented), can be cost effectively used for snubber replacement programs, mitigation of hydraulic transients, pipe whip and as a thermal stop. The device has pre-set gaps to allow free thermal movement. During a seismic or other dynamic load event, if the pipe movement exceeds the gap dimension, the device acts as an elastic or elastic-plastic restraint. The device also has a unique design feature for not exceeding the restraint force beyond a specified limit design value. To analyze piping systems with gap supports having elastic-plastic characteristics, modal analysis procedures for both response spectrum and time history methods are developed. The comparison of responses obtained from the procedures with nonlinear time history analysis and test results available in the literature shows excellent correlation. A pilot program conducted for snubber replacement with E-BARs demonstrates that the limit force feature of E-BAR makes them very attractive for snubber replacement. This is because a particular E-BAR with a specified limit design force can be selected, such that, the E-BAR replacing the snubber does not require any modifications be made to the existing support steel and hardware.