Conformational stability of factor VIIa: biophysical studies of thermal and guanidine hydrochloride-induced denaturation

The binding of the multidomain protein factor VIIa (fVIIa) to tissue factor provides the interprotein communication necessary to make fVIIa an efficient catalyst of the initial event in the extrinsic pathway of blood coagulation. We have investigated the stability of individual domains in fVIIa and the influence of Ca2+ and an irreversible active-site inhibitor (FFR-chloromethyl ketone). Equilibrium guanidine hydrochloride (GuHCl)-induced unfolding monitored by tryptophan fluorescence and far-UV circular dichroism (CD) demonstrated that the gamma-carboxyglutamic acid (Gla) domain unfolds at 0.3 M GuHCl and the serine protease (SP) domain at 3 M GuHCl and that Ca2+ is a prerequisite for the formation of an ordered, compact structure in the Gla domain. The loss of amidolytic activity coincides with the first transition, which is stabilized by the active-site inhibitor, and a change in the environment of the active site is demonstrated using a fluorescent inhibitor (DEGR-chloromethyl ketone). Thermal unfolding monitored by differential scanning calorimetry (DSC) reveals that Ca2+ stabilizes the SP domain slightly, increasing the unfolding temperature by 2.7 degrees C. In addition, Ca2+ is required for a large enthalpy change concomitant with unfolding of the Gla domain, and this unfolding enthalpy is only detectable in the presence of the SP domain, indicating some kind of interaction between these domains. Thermal unfolding measured by CD indicates secondary structural changes at the same temperature as the heat absorption in the DSC but only when both the Gla domain and the SP domain are present together with Ca2+ ions. Taken together, these results indicate a Ca2+-dependent interaction between the Gla domain and the SP domain, implying a high degree of flexibility of the domains in free fVIIa. It is also shown that the epidermal growth factor-like domains are stable at elevated temperatures and high GuHCl concentrations. Moreover, already at physiological temperature, subtle structural changes take place which influence the overall shape of fVIIa and are detrimental to its enzymatic activity.     

OWL2Vec*: embedding of OWL ontologies

Machine Learning - Semantic embedding of knowledge graphs has been widely studied and used for prediction and statistical analysis tasks across various domains such as Natural Language Processing...     

Agile distributed software development: enacting control through media and context

While face‐to‐face interaction is fundamental in agile software development, distributed environments must rely extensively on mediated interactions. Practicing agile principles in distributed environments therefore poses particular control challenges related to balancing fixed vs. evolving quality requirements and people vs. process‐based collaboration. To investigate these challenges, we conducted an in‐depth case study of a successful agile distributed software project with participants from a Russian firm and a Danish firm. Applying Kirsch's elements of control framework, we offer an analysis of how control was enacted through the project context and in the participants' mediated communication. The analysis reveals that formal measurement and evaluation control were persistently enacted through mediated communication. These formal control practices were, however, predominantly carried out in conjunction with informal roles and relationships such as clan‐like control inherent in agile development. Overall, the study demonstrates that, if appropriately applied, communication technologies can significantly support distributed, agile practices by allowing concurrent enactment of both formal and informal controls. The paper discusses these findings as they relate to previous research and concludes with their implications for future research.     

Effect of high temperature heat treatment on intergranular corrosion of AlMgSi(Cu) model alloy

Copper containing 6000-series aluminium alloys may become susceptible to intergranular corrosion (IGC) as a result of improper thermomechanical processing. Effect of cooling rate after solution heat treatment on the corrosion behaviour of a model AlMgSi(Cu) alloy of nominal composition (wt%) 0.6 Mg, 0.6 Si, 0.2 Fe, 0.2 Mn and 0.1 Cu was investigated. Slow cooling rates were simulated by isothermal treatment for predetermined times in lower temperature baths immediately after solution heat treatment. Treatment for 10-100 s at temperatures below 400 °C introduced susceptibility to IGC. Longer heat treatment at the same temperatures introduced susceptibility to pitting. A corrosion resistant time zone was found between the zones of IGC and pitting at temperatures lower than 350 °C. Quenching in water after solution heat treatment prevented IGC. IGC was related to microgalvanic coupling between the noble Q-phase (Al₄Mg₈Si₇Cu₂) grain boundary precipitates and the adjacent depleted zone. Pitting was attributed to coarse particles in the matrix. Possible mechanisms causing the corrosion resistant intermediate zone are discussed. The results indicate possible methods for obtaining increased corrosion resistance of similar alloys by proper thermal processing.     

Near-Surface Deformation in an Alumina–Silicon Carbide-Whisker Composite due to Surface Machining

Deformation due to two different surface-machining conditions—grinding (126 μm diamond) and polishing (3 μm diamond)—in an uniaxial hot-pressed Al₂O₃–30%-SiC-whisker composite has been investigated. A Warren–Averbach analysis of grazing incidence X-ray diffractometry data shows that the deformation is localized to the very top surface zone. The cell size and the root mean square of the strain show a gradient in the deformed layer. Transmission electron microscopy studies, in cross-sectional view, also show a near-surface deformation zone containing dislocations, twins, and cracks. This is seen for both machining procedures, but the depth of the zone and the degree of deformation, in terms of dislocation density and number of cracks, is much higher in the roughly ground specimen than in the polished one. For comparison, a monolithic Al₂O₃ sample also has been studied after grinding. The deformation zone is very similar to the Al₂O₃–SiC sample with the same grinding condition, but cracks and dislocations are present at a slightly larger depth. The deformation depth for the polished Al₂O₃–SiC sample is ∼50 nm. In the ground Al₂O₃–SiC sample, the deformation depth is 1–1.5 μm and corresponds to the grain size. The deformation zone in the ground monolithic Al₂O₃ sample is 1.5–2 μm deep. The observed grain-boundary cracks are almost parallel to the surface and may originate from nonaccommodated plastic flow between grains.     

Manipulability of leader–follower networks with the rigid-link approximation

This paper introduces the notion of manipulability to mobile, multi-agent networks as a tool to analyze the instantaneous effectiveness of injecting control inputs at certain, so-called leader nodes in the network. Effectiveness is interpreted to characterize how the movements of the leader nodes translate into responses among the remaining follower nodes. This notion of effectiveness is a function of the interaction topologies, the agent configurations, and the particular choice of inputs used to influence the network. In fact, classic manipulability is an index used in robotics to analyze the singularity and efficiency of configurations of robot-arm manipulators. To define similar notions for leader–follower networks, we use a rigid-link approximation of the follower dynamics and, under this assumption, we prove that the instantaneous follower velocities can be uniquely determined from that of the leaders’, which allows us to define a meaningful and computable manipulability index for the leader–follower networks. This paper examines the property of the proposed index in simulation and with real mobile robots, and demonstrates how the index can be used to find effective interaction topologies.     

Influence of the type of reducing agent (H2, CO,C3H6 and C3H8) on the reduction of stored NO X in a Pt/BaO/Al2O3 model catalyst

In this investigation, a comparative study for a NO _X storage catalytic system was performed focusing on the parameters that affect the reduction by using different reductants (H₂, CO, C₃H₆ and C₃H₈) and different temperatures (350, 250 and 150 °C), for a Pt/BaO/Al₂O₃ catalyst. Transient experiments show that H₂ and CO are highly efficient reductants compared to C₃H₆ which is somewhat less efficient. H₂ shows a significant reduction effect at relatively low temperature (150 °C) but with a low storage capacity. We find that C₃H₈ does not show any NO _X reduction ability for NO _X stored in Pt/BaO/Al₂O₃ at any of the temperatures. The formation of ammonia and nitrous oxide is also discussed.

     

Surface properties of surfactants derived from natural products. Part 1: Syntheses and structure/property relationships—Solubility and emulsification

Several novel and some previously known, mostly sugar-based, surfactants have been synthesized and some of their surface properties have been characterized and compared with those of commercial nonylphenol ethoxylates. The surfactant solubility in water, ethanol, and dodecane was studied. The properties of these compounds as emulsification agents in systems composed of the surfactant with water/isopropyl myristate, water/rapeseed oil, and water/dodecane are presented. The aqueous solubility of the surfactants follows the general trend expected from their hydrophilic-lipophilic balance according to Griffin (HLBG), but it is also clear that the nature of the headgroup and the structure of the nonpolar part affect the solubility in a manner not aptured in the standard HLBG concept. An ester or amine group as the connecting unit between the hydrophile and the hydrophobe produces a more water-soluble surfactant than the corresponding amide derivative. Some effective emulsifiers were found. For instance, the surfactants with a dehydroabietic nonpolar group appear to be promosing emulsifiers. Most sugar-based surfactants were able to form macroemulsions of up to around 2 wt/vol% of oil. The stability of many of these amulsions was very high, extending for months.     

Enantiomeric Composition of Monoterpene Hydrocarbons in Some Conifers and Receptor Neuron Discrimination of α-Pinene and Limonene Enantiomers in the Pine Weevil, Hylobius abietis

The enantiomeric composition of seven monoterpene hydrocarbons in headspace volatiles of spruce sawdust and seedlings (Picea abies), pine seedlings (Pinus sylvestris), and branches of juniper (Juniperus communis) was determined by gas chromatographic separation on a -Cyclodextrin column. For the six monoterpenes, -pinene, camphene, -pinene, sabinene, limonene, and -phellandrene, both enantiomers were present, whereas for 3-carene only the (+)-configuration was found. The amount of each enantiomer varied considerably both in relation to total amount of all of them, and for the six pairs also in relation to the opposite enantiomer. One olfactory receptor neuron in the pine weevil (Hylobius abietis) showed a strong response to -pinene when stimulated with all four headspace materials via a GC equipped with a DB-WAX column. The same neuron was subsequently tested with repeated stimulations via the GC effluent containing the (+)- or (–)-enantiomer. A marked better response to (+)- than to (–)--pinene was elicited. Another olfactory receptor neuron that responded strongly to limonene when stimulated with the spruce volatiles was tested for enantiomers of limonene. This neuron responded more strongly to (–)- than to (+)-limonene, when stimulated alternately with each of the limonene enantiomers. Discrimination between enantiomers by plant olfactory receptor neurons suggests that the enantiomeric ratios of volatile compounds may be important in host location by the pine weevil.     

Sulfur deactivation and regeneration of Pt/BaO/Al2O3 and Pt/SrO/Al2O3 NO x storage catalysts

Transient experiments were performed to study sulfur deactivation and regeneration of Pt/BaO/Al₂O₃ and Pt/SrO/Al₂O₃ NO _x storage catalysts. It was found that the strontium-based catalysts are more easily regenerated than the barium-based catalysts and that a higher fraction of the NO _x storage sites are regenerated when H₂ is used in combination with CO₂ compared to H₂ only.