Determination of the pKa values of titratable groups of an antigen-antibody complex, HyHEL-5-hen egg lysozyme

The titration behavior of the ionizable residues of the HyHEL-5–henegg lysozyme complex and its individual components has beenstudied using continuum electrostatic calculations. Severalresidues of HyHEL-5 had pK_a values shifted away from model valuesfor isolated residues by more than three pH units. Shifts awayfrom the model values were smaller for the residues of hen egglysozyme. A moderate variation in the pK_a values of the titratablegroups was observed upon increase of the ionic strength from0 to 100 mM, amounting to 1–2 pH units in most cases.Under physiological conditions, the net charge of HyHEL-5 wasopposite that for hen egg lysozyme. Several residues, includingthose involved in the Arg–Glu salt bridges that have beenproposed to be important in antibody-antigen binding, had pK_avalues that were changed significantly upon binding. The maintitration event upon antibody-antigen binding appears to beloss of a proton from residue GluH50 of the Fv molecule. Thelimitations of our calculation methods and the role they mightplay in the design of antibodies for use in assays, sensorsand separations are discussed     

Two-dimensional correlation relaxometry studies of cement pastes performed using a new one-sided NMR magnet

We present preliminary results of the first NMR T₁-T₂ two-dimensional relaxation correlation experiments performed using a one-sided NMR system in cement based materials. Two-dimensional correlation relaxometry has itself only recently been demonstrated in cement paste where it proved to be a particularly sensitive probe of pore-water dynamics providing direct information on exchange of water between the gel and capillary pore networks. Further to this we have observed differences in the structural development of a selection of cement pastes throughout the early stages of hydration and verified the theoretical frequency dependence of the ratio T₁ / T₂. When coupled with instrumentation developments in one-sided NMR magnets the way is opened to detailed, spatially resolved studies of the development of hydration and porosity in the surface layers (top 50 mm) of cementitious materials. A new magnet, suitable for such applications, is discussed.     

Accumulation of eicosapentaenoic acid in plasma phospholipids of subjects fed canola oil

The metabolism of α-linolenic acid from canola oil was studied in eight normolipidemic men. The 42-day study was divided into three periods: a 6-day pre-experimental and two 18-day experimental. Approximately 75% of the dietary fat (28% of total energy) was provided by a mixture of fats during the pre-experimental period and either canola oil (CO) or sunflower oil (SO) during the experimental periods. The CO and SO diets were fed in a cross-over design. The ratios of linoleic to linolenic acid were 2.6∶1 and 73.9∶1 in the CO and SO diets, respectively. Dietary fat source had an effect on plasma phospholipid fatty acids: 18∶1n−9, 18∶3n−3 and 20∶5n−3 were higher (p<0.05), and 18∶2n−6 was lower in the phosphatidylcholine fraction; 18∶1n−9 was higher and 20∶4n−6 lower in the phosphatidyl-ethanolamine fraction; and 18∶1n−9 and 20∶5n−3 were higher and 20∶4n−6 and 22∶6n−3 were lower in the alkenylacyl-ethanolamine phospholipid fraction on the CO diet as compared to the SO diet. Consumption of the canola oil diet resulted in higher n−3 fatty acid levels and lower n−6 fatty acid levels in plasma phospholipids than consumption of the sunflower oil diet.     

Dual Verbal Elicitation: The Complementary Use of Concurrent and Retrospective Reporting Within a Usability Test

Verbal protocols are the primary tool for understanding users' task-solving behaviors during usability testing. A qualitative study that examined the utility of combining a concurrent and retrospective think-aloud within the same usability test is described. The results indicate that although there was significant overlap between the types of utterances produced during each think-aloud, the retrospective phase produced more verbalizations that were relevant to usability analysis, for example, helpful self-assessments of performance, yielding insights into the impact of encountered difficulties. However, a small number of less desirable utterance types emerged: hypothesising, rationalizing, and forgetting. When used together, both methods contributed to an understanding of usability issues; the concurrent phase yielded more usability issues overall, and the retrospective data improved the understanding of these by (a) reinforcement: users highlighted the impact of an issue on their experience, (b) elaboration: users would provide causal explanations of encountered difficulties, and (c) context: users provided information about the product's context of use.     

The Impact of Interaction and Social Learning on Aggregate Expectations

We examine the effect of information sharing within small world networks. Agents receive a signal correlated with the state of the world (SoW) which is adjusted following discussions with neighbours. If one agent in the network, referred to as an expert, does not engage in social learning (that is they always follow their own signal) then all agents learn the SoW. It is found that volatility in the mean level of expectations varies with changes in the number of experts and the network structure. A trade-off emerges between the level of volatility and the speed at which agents learn of changes to the SoW. A second finding is that certain network structures lead to information cascades.      

Cell surface chondroitin sulfate proteoglycans in tumor cell adhesion, motility and invasion

Genes that encode enzymes that convert inactive "prodrugs" into anticancer metabolites may be therapeutically useful against brain tumors. Unlike other genes tested to date in brain tumor models, the Escherichia coli gpt gene is unique in that it not only sensitizes cells to the prodrug 6-thioxanthine (6TX) but also encodes resistance to a different regimen (mycophenolic acid, xanthine, and hypoxanthine), thus providing a means to select for gpt-positive cells. In the present study, rat C6 glioma cells were infected with a retrovirus vector that transduces this gene. A clonal line (C6GPT-7) was derived that exhibited significant 6TX susceptibility in vitro with an ID50 of 2.5 mumol/L, whereas 50% growth inhibition of parental C6 cells was not achieved at concentrations tested (up to 50 mumol/L). This line also exhibited significant sensitivity to 6-thioguanine (6TG), with an ID50 of 0.05 mumol/L, whereas 50% growth inhibition of parental C6 cells was achieved at 0.5 mumol/L. In a "bystander" assay, C6GPT-7 tumor cells efficiently transferred 6TX sensitivity to C6 cells at ratios as low as 1:9 (C6GPT-7:C6). This in vitro bystander effect was abrogated when C6GPT-7 and C6 cells were separated by a microporous membrane, suggesting that it was not mediated by highly diffusible metabolites. In vivo both 6TX and 6TG significantly inhibited the growth of subcutaneously transplanted C6GPT-7 cells but not that of C6 cells in athymic mice. In an intracerebral model, both 6TX and 6TG exhibited significant antiproliferative effects against tumors formed by C6GPT-7 cells. These findings provide a basis for exploring further gene therapy strategies based on in vivo transfer of the E coli gpt gene to provide chemosensitivity against 6TX and 6TG.     

Reflections on the evolution of implicit Navier–Stokes algorithms

This article gives a historical perspective on the evolution of implicit algorithms for the Navier–Stokes equations that utilize time or Newton linearizations and various forms of approximate factorization including alternating-direction, lower/upper, and symmetric relaxation schemes. A theme of the paper is how progress in implicit Navier–Stokes algorithms has been influenced and enabled by the introduction of characteristic-based upwind approximations, unstructured-grid discretizations, parallelization, and by advances in computer performance and architecture. Historical examples of runtime, problem size, and estimated cost are given for actual and hypothetical Navier–Stokes flow cases from the past 40 years.     

Time-correlated single photon counting FLIM: some considerations for physiologists

Recent developments in cellular imaging now permit the minimally invasive study of protein interactions in living cells. These advances are of enormous interest to cell biologists, as proteins rarely act in isolation, but rather in concert with others in forming cellular machinery. Up until recently, all protein interactions had to be determined in vitro using biochemical approaches. This biochemical legacy has provided cell biologists with the basis to test defined protein-protein interactions not only inside cells, but now also with spatial resolution. More recent developments in TCSPC imaging are now also driving towards being able to determine protein interaction rates with similar spatial resolution, and together, these experimental advances allow investigators to perform biochemical experiments inside living cells. Here, we discuss some findings we have made along the way which may be useful for physiologists to consider.     

Classification of abnormal plant operation using multiple process variable trends

This paper illustrates two strategies for the detection and classification of abnormal process operating conditions in which multiple process variable trends are available. The first strategy uses a hidden Markov model (HMM) for overall process classification while the second method uses a back-propagation neural network (BPNN) to determine the overall process classification. The methods are compared in terms of their ability to detect and correctly diagnose a variety of abnormal operating conditions for a non-isothermal CSTR simulation. For the case study problem, the BPNN method resulted in better classification accuracy with a moderate increase in training time compared with the HMM approach.