Quantifying the Differences in Binding Affinity of Reduced Glutathione for Glutathione S-Transferase at pH 6.5 and 8.5 Using Isothermal Titration Calorimetry

The binding affinity between an enzyme and its substrate is often dependent on the pH of the local environment. Knowing the pH at which reduced glutathione (GSH) binds with the highest affinity to the enzyme glutathione S-transferase (GST) is useful for determining the optimal pH for purification of GST-fusion proteins during GST-affinity chromatography. In this study, GST of the species Schistosoma japonicum was purified, quantified, and utilized to study its binding interaction with GSH at pH 6.5 and 8.5 via isothermal titration calorimetry (ITC). After protein expression, extraction, and purification, the GST concentration was quantified using QubitTM fluorometry. Thermodynamic properties and a dissociation constant (KD) for each experiment were obtained utilizing the MicroCal PEAQ-ITC Analysis Software for the binding of GSH to GST at pH 6.5 and 8.5. Statistical analysis of the technical replicate data was performed to obtain an average and standard deviation of the KD at each pH point. The results indicate a statistically significant difference (

Effect of high oxygen pressure annealing on superconducting Nd1.85Ce0.15CuO4 thin films by pulsed laser deposition from Cu-enriched targets

We show that the quality of Nd1.85Ce0.15CuO4 films grown by pulsed laser deposition can be enhanced by using a non-stoichiometric target with extra copper added to suppress the formation of a parasitic (Nd, Ce)2O3 phase. The properties of these films are less dependent on the exact annealing procedure after deposition as compared to films grown from a stoichiometric target. Film growth can be followed by a 1 bar oxygen annealing, after an initial vacuum annealing, while retaining the superconducting properties and quality. This enables the integration of electron-doped cuprates with their hole-doped counterparts on a single chip, to create, for example, superconducting pn-junctions.Comment: This is an author-created, un-copyedited version of an article accepted for publication in Superconductor Science and Technology. The publisher is not responsible for any errors or omissions in this version of the manuscript or any version derived from it. The Version of Record is available online at http://dx.doi.org/10.1088/0953-2048/27/4/04401

A parameterized approach to modeling and forecasting mortality

A new method is proposed of constructing mortality forecasts. This parameterized approach utilizes Generalized Linear Models (GLMs), based on heteroscedastic Poisson (non-additive) error structures, and using an orthonormal polynomial design matrix. Principal Component (PC) analysis is then applied to the cross-sectional fitted parameters. The produced model can be viewed either as a one-factor parameterized model where the time series are the fitted parameters, or as a principal component model, namely a log-bilinear hierarchical statistical association model of Goodman [Goodman, L.A., 1991. Measures, models, and graphical displays in the analysis of cross-classified data. J. Amer. Statist. Assoc. 86(416), 1085–1111] or equivalently as a generalized Lee–Carter model with p interaction terms. Mortality forecasts are obtained by applying dynamic linear regression models to the PCs. Two applications are presented: Sweden (1751–2006) and Greece (1957–2006)

Live imaging of tumor initiation in zebrafish larvae reveals a trophic role for leukocyte-derived PGE(2)

SummaryEpidemiology studies and clinical trials have suggested that the use of non-steroidal anti-inflammatory drugs (NSAIDs), including aspirin, can significantly reduce the incidence of and mortality associated with many cancers [1–3], and upregulation of the COX2-PGE2 pathway in tumor microenvironments might drive several aspects of cancer progression [4–6]. For these reasons, the mechanisms linking COX blockade and cancer prevention have long been an area of active investigation [7]. During carcinogenesis, COX-2 is expressed both by malignant epithelial cells [8, 9] and by tumor-associated stromal cells, including macrophages [10–12], but the observation that NSAIDs are most effective in cancer prevention in APCmin/+ mice if the mice are treated from conception [13] suggests that the COX-2/PGE2 pathway might also be critical at the earliest stages of tumor development. In this study we take advantage of the translucency and genetic tractability of zebrafish larvae to investigate the involvement of inflammatory cells at cancer initiation, when transformed cells first arise in tissues. We previously showed that innate immune cells supply early transformed cells with proliferative cues [14] and, by using complementary pharmacological and genetic experiments, we now show that prostaglandin E2 (PGE2) is the trophic signal required for this expansion of transformed cells. Our in vivo observations at these early stages of cancer initiation provide a potential mechanistic explanation for why long-term use of low doses of NSAIDs, including aspirin, might reduce cancer onset

Can the British Heart Foundation PocketCPR Application Improve the Performance of Chest Compressions During Bystander Resuscitation: a Randomised Crossover Manikin Study

This study aims to determine whether the British Heart Foundation (BHF) PocketCPR application can improve the depth and rate of chest compression, and therefore be confidently recommended for bystander use. 118 candidates were recruited into a randomised crossover manikin trial. Each candidate performed CPR for two-minutes without instruction, or performed chest compressions using the PocketCPR application. Candidates then performed a further two minutes of CPR within the opposite arm. The number of chest compressions performed improved when PocketCPR was used compared to chest compressions when it was not (44.28% v40.57, P<0.001). The number of chest compressions performed to the required depth was higher in the PocketCPR group (90.86 v 66.26). The BHF PocketCPR application improved the percentage of chest compressions that were performed to the required depth. Despite this, more work is required in order to develop a feedback device that can improve bystander CPR without creating delay

Common mortality modeling and coherent forecasts. An empirical analysis of worldwide mortality data

A new common mortality modeling structure is presented for analyzing mortality dynamics for a pool of countries, under the framework of generalized linear models (GLM). The countries are first classified by fuzzy c-means cluster analysis in order to construct the common sparse age-period model structure for the mortality experience. Next, we propose a method to create the common sex difference age-period model structure and then use this to produce the residual age-periodmodel structure for each country and sex. The time related principal components are extrapolated using dynamic linear regression (DLR) models and coherent mortality forecasts are investigated. We make use of mortality data from the “Human Mortality Database”

Proximity-to-goal as a constraint on patterns of behaviour in attacker-defender dyads in team games

Abstract The aim of this study was to determine whether spatiotemporal interactions between footballers and the ball in 1 vs. 1 sub-phases are influenced by their proximity to the goal area. Twelve participants (age 15.3 +/- 0.5 years) performed as attackers and defenders in 1 vs. 1 dyads across three field positions: (a) attacking the goal, (b) in midfield, and (c) advancing away from the goal area. In each position, the dribbler was required to move beyond an immediate defender with the ball towards the opposition goal. Interactions of attacker-defender dyads were filmed with player and ball displacement trajectories digitized using manual tracking software. One-way repeated measures analysis of variance was used to examine differences in mean defender-to-ball distance after this value had stabilized. Maximum attacker-to-ball distance was also compared as a function of proximity-to-goal. Significant differences were observed for defender-to-ball distance between locations (a) and (c) at the moment when the defender-to-ball distance had stabilized (a: 1.69 +/- 0.64 m; c: 1.15 +/- 0.59 m; P < 0.05). Findings indicate that proximity-to-goal influenced the performance of players, particularly when attacking or advancing away from goal areas, providing implications for training design in football. In this study, the task constraints of football revealed subtly different player interactions than observed in previous studies of dyadic systems in basketball and rugby union

Characterisation of potato crisp effective porosity using micro-CT

Background The effective porosity is an important quantitative parameter for food products that has a significant effect on taste and quality. It is challenging to quantify the apparent porosity of fried potato crisps as they have a thin irregularly shaped cross section containing oil and water. This study uses a novel micro-CT technique to determine the solid volume fraction and hence the effective porosity of three types of potato crisps: standard continuously fried crisps, microwaved crisps, and continuously fried ‘kettle’ crisps. Results It was found that continuously fried kettle crisps had the lowest effective porosity at 0.54, providing the desired crunchy taste and lower oil contents. Crisps produced using a microwave process designed to mimic the dehydration process of standard continuous fried crisps had an effective porosity of 0.65, which was very similar to the effective porosity of 0.63 for standard continuously fried crisps. The results were supported by the findings of a forced preference consumer test. Conclusion The effective porosity affects the product taste and is therefore a critical parameter. This study shows that micro-CT analysis can be used to characterise the change in effective porosity of a thin irregularly shaped food product, caused by a change of cooking procedure

The effect of exercise on innate mucosal immunity

METHODS The authors conducted a prospective observational study comparing salivary lactoferrin and lysozyme concentration over 5 months (chronic changes) in elite rowers (n=17, mean age 24.3+/-4.0 years) with sedentary individuals (controls) (n=18, mean age=27.2+/-7.1 years) and a graded exercise test to exhaustion (acute changes) with a cohort of elite rowers (n=11, mean age 24.7+/-4.1). RESULTS Magnitudes of differences and changes were interpreted as a standardised (Cohen's) effect size (ES). Lactoferrin concentration in the observational study was approximately 60% lower in rowers than control subjects at baseline (7.9+/-1.2 microg/ml mean+/-SEM, 19.4+/-5.6 microg/ml, p=0.05, ES=0.68, 'moderate') and at the midpoint of the season (6.4+/-1.4 microg/ml mean +/- SEM, 21.5+/-4.2 microg/ml, p=0.001, ES=0.89, 'moderate'). The concentration of lactoferrin at the end of the study was not statistically significant (p=0.1) between the groups. There was no significant difference between rowers and control subjects in lysozyme concentration during the study. There was a 50% increase in the concentration of lactoferrin (p=0.05, ES=1.04, 'moderate') and a 55% increase in lysozyme (p=0.01, ES=3.0, 'very large') from pre-exercise to exhaustion in the graded exercise session. CONCLUSION Lower concentrations of these proteins may be indicative of an impairment of innate protection of the upper respiratory tract. Increased salivary lactoferrin and lysozyme concentration following exhaustive exercise may be due to a transient activation response that increases protection in the immediate postexercise period