Differential recognition of Staphylococcus aureus quorum-sensing signals depends on both extracellular loops 1 and 2 of the transmembrane sensor AgrC.

Virulence in Staphylococcus aureus is regulated via agr-dependent quorum sensing in which an autoinducing peptide (AIP) activates AgrC, a histidine protein kinase. AIPs are usually thiolactones containing seven to nine amino acid residues in which the thiol of the central cysteine is linked to the alpha-carboxyl of the C-terminal amino acid residue. The staphylococcal agr locus has diverged such that the AIPs of the four different S. aureus agr groups self-activate but cross-inhibit. Consequently, although the agr system is conserved among the staphylococci, it has undergone significant evolutionary divergence whereby to retain functionality, any changes in the AIP-encoding gene (agrD) that modifies AIP structure must be accompanied by corresponding changes in the AgrC receptor. Since AIP-1 and AIP-4 only differ by a single amino acid, we compared the transmembrane topology of AgrC1 and AgrC4 to identify amino acid residues involved in AIP recognition. As only two of the three predicted extracellular loops exhibited amino acid differences, site-specific mutagenesis was used to exchange the key AgrC1 and AgrC4 amino acid residues in each loop either singly or in combination. A novel lux-based agrP3 reporter gene fusion was constructed to evaluate the response of the mutated AgrC receptors. The data obtained revealed that while differential recognition of AIP-1 and AIP-4 depends primarily on three amino acid residues in loop 2, loop 1 is essential for receptor activation by the cognate AIP. Furthermore, a single mutation in the AgrC1 loop 2 resulted in conversion of (Ala5)AIP-1 from a potent antagonist to an activator, essentially resulting in the forced evolution of a new AIP group. Taken together, our data indicate that loop 2 constitutes the predicted hydrophobic pocket that binds the AIP thiolactone ring while the exocyclic amino acid tail interacts with loop 1 to facilitate receptor activation.

Pseudomonas syringae pv. coryli, the causal agent of bacterial twig dieback of Corylus avellana

Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by a polyphasic approach. All strains were assessed by fatty acids analysis and repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX and ERIC primer sets. Representative strains also were assessed by sequencing the 16S rDNA and hrpL genes, determining the presence of the syrB gene, testing their biochemical and nutritional characteristics, and determining their pathogenicity to hazelnut and other plants species or plant organs. Moreover, they were compared with reference strains of other phytopathogenic pseudomonads. The strains from hazelnut belong to Pseudomonas syringae (sensu latu), LOPAT group Ia. Both fatty acids and repetitive-sequence-based PCR clearly discriminate such strains from other Pseudomonas spp., including P. avellanae and other P. syringae pathovars as well as P. syringae pv. syringae strains from hazelnut. Also, the sequencing of 16S rDNA and hrpL genes differentiated them from P. avellanae and from P. syringae pv. syringae. They did not possess the syrB gene. Some nutritional tests also differentiated them from related P. syringae pathovars. Upon artificial inoculation, these strains incited severe twig diebacks only on hazelnut. Our results justify the creation of a new pathovar because the strains from hazelnut constitute a homogeneous group and a discrete phenon. The name of P. syringae pv. coryli is proposed and criteria for routine identification are presented.

Dynamic simulation for closed loops in heating systems with decision-making support

Nowadays utilising the proper HVAC system is essential both in extreme weather conditions and dense buildings design. Hydraulic loops are the most common parts in all air conditioning systems. This article aims to investigate the performance of different hydraulic loop arrangements in variable flow systems. Technical, economic and environmental assessments have been considered in this process. A dynamic system simulation is generated to evaluate the system performance and an economic evaluation is conducted by whole life cost assessment. Moreover, environmental impacts have been studied by considering the whole life energy consumption, CO2 emission, the embodied energy and embodied CO2 of the system components. Finally, decision-making in choosing the most suitable hydraulic system among five well-known alternatives has been proposed.

The causal surface and its effect on distribution transparency in a distributed virtual environment

One goal in the development of distributed virtual environments (DVEs) is to create a system such that users are unaware of the distribution-the distribution should be transparent. The paper begins by discussing the general issues in DVEs that might make this possible, and a system that allows some level of distribution transparency is described. The system described suffers from effects of inconsistency, which in turn cause undesirable visual effects. The causal surface is introduced as a solution that removes these visual effects. The paper then introduces two determining factors of distribution transparency relating to user perception and performance. With regard to these factors, two hypotheses are stated relating to the causal surface. A user-trial on forty-five subjects is used to validate the hypotheses. A discussion of the results of the trial concludes that the causal surface solution does significantly improve the distribution transparency in a DVE.

Causal volumes in distributed virtual reality

Research to date has tended to concentrate on bandwidth considerations to increase scalability in distributed interactive simulation and virtual reality systems. This paper proposes that the major concern for latency in user interaction is that of the fundamental limit of communication rate due to the speed of light. Causal volumes and surfaces are introduced as a model of the limitations of causality caused by this fundamental delay. The concept of virtual world critical speed is introduced, which can be determined from the causal surface. The implications of the critical speed are discussed, and relativistic dynamics are used to constrain the object speed, in the same way speeds are bounded in the real world.

Maximising concurrency and scalability in a consistent, causal, distributed virtual reality system, whilst minimising the effect of network delays

The development of large scale virtual reality and simulation systems have been mostly driven by the DIS and HLA standards community. A number of issues are coming to light about the applicability of these standards, in their present state, to the support of general multi-user VR systems. This paper pinpoints four issues that must be readdressed before large scale virtual reality systems become accessible to a larger commercial and public domain: a reduction in the effects of network delays; scalable causal event delivery; update control; and scalable reliable communication. Each of these issues is tackled through a common theme of combining wall clock and causal time-related entity behaviour, knowledge of network delays and prediction of entity behaviour, that together overcome many of the effects of network delay.

Maximising concurrency and scalability in a consistent, causal, distributed virtual reality system, whilst minimising the effect of network delays

The development of large scale virtual reality and simulation systems have been mostly driven by the DIS and HLA standards community. A number of issues are coming to light about the applicability of these standards, in their present state, to the support of general multi-user VR systems. This paper pinpoints four issues that must be readdressed before large scale virtual reality systems become accessible to a larger commercial and public domain: a reduction in the effects of network delays; scalable causal event delivery; update control; and scalable reliable communication. Each of these issues is tackled through a common theme of combining wall clock and causal time-related entity behaviour, knowledge of network delays and prediction of entity behaviour, that together overcome many of the effects of network delays.

Causal evidence for frontal involvement in memory target maintenance by posterior brain areas during distracter interference of visual working memory

Dorsolateral prefrontal cortex (DLPFC) is recruited during visual working memory (WM) when relevant information must be maintained in the presence of distracting information. The mechanism by which DLPFC might ensure successful maintenance of the contents of WM is, however, unclear; it might enhance neural maintenance of memory targets or suppress processing of distracters. To adjudicate between these possibilities, we applied time-locked transcranial magnetic stimulation (TMS) during functional MRI, an approach that permits causal assessment of a stimulated brain region's influence on connected brain regions, and evaluated how this influence may change under different task conditions. Participants performed a visual WM task requiring retention of visual stimuli (faces or houses) across a delay during which visual distracters could be present or absent. When distracters were present, they were always from the opposite stimulus category, so that targets and distracters were represented in distinct posterior cortical areas. We then measured whether DLPFC-TMS, administered in the delay at the time point when distracters could appear, would modulate posterior regions representing memory targets or distracters. We found that DLPFC-TMS influenced posterior areas only when distracters were present and, critically, that this influence consisted of increased activity in regions representing the current memory targets. DLPFC-TMS did not affect regions representing current distracters. These results provide a new line of causal evidence for a top-down DLPFC-based control mechanism that promotes successful maintenance of relevant information in WM in the presence of distraction.

Intrafirm causal ambiguity in an international context

This paper examined the incidence of intrafirmcausalambiguity in the management's perception concerning the critical drivers of their firms’ performance. Building on insights from the resource-based view we developed and tested hypotheses that examine (1) linkage ambiguity as a discrepancy between perceived and measured resource–performance linkages, (2) characteristic ambiguity for resources and capabilities with a high degree of complexity and tacitness, and (3) the negative association between linkage ambiguity and performance. The observations based on the explicit perceptions of 356 surveyed managers were contrasted with the empirical findings of the resource/performance relationship derived by structural equation modelling from the same data sample. The findings validate the presence of linkage ambiguity particularly in the case of resources and capabilities with higher degree of characteristic ambiguity. The findings also provide empirical evidence in support of the advocacy for a negative relationship between intrafirmcausalambiguity and performance. The paper discusses the potential reasons for the disparities between empirical findings and management's perceptions of the key determinants of export success and makes recommendations for future research.

Steering without Circe : attending to reinforcing loops in social systems

Relating system dynamics to the broad systems movement, the key notion is that reinforcing loops deserve no less attention than balancing loops. Three specific propositions follow. First, since reinforcing loops arise in surprising places, investigations of complex systems must consider their possible existence and potential impact. Second, because the strength of reinforcing loops can be misinferred - we include an example from the field of servomechanisms - computer simulation can be essential. Be it project management, corporate growth or inventory oscillation, simulation helps to assess consequences of reinforcing loops and options for interventions. Third, in social systems the consequences of reinforcing loops are not inevitable. Examples concerning globalization illustrate how difficult it might be to challenge such assumptions. However, system dynamics and ideas from contemporary social theory help to show that even the most complex social systems are, in principle, subject to human influence. In conclusion, by employing these ideas, by attending to reinforcing as well as balancing loops, system dynamics work can improve the understanding of social systems and illuminate our choices when attempting to steer them.

Formal theory building for the avalanche game: explaining counter-intuitive behaviour of a complex system using geometrical and human behavioural/physiological effects

In 'Avalanche', an object is lowered, players staying in contact throughout. Normally the task is easily accomplished. However, with larger groups counter-intuitive behaviours appear. The paper proposes a formal theory for the underlying causal mechanisms. The aim is to not only provide an explicit, testable hypothesis for the source of the observed modes of behaviour-but also to exemplify the contribution that formal theory building can make to understanding complex social phenomena. Mapping reveals the importance of geometry to the Avalanche game; each player has a pair of balancing loops, one involved in lowering the object, the other ensuring contact. For more players, sets of balancing loops interact and these can allow dominance by reinforcing loops, causing the system to chase upwards towards an ever-increasing goal. However, a series of other effects concerning human physiology and behaviour (HPB) is posited as playing a role. The hypothesis is therefore rigorously tested using simulation. For simplicity a 'One Degree of Freedom' case is examined, allowing all of the effects to be included whilst rendering the analysis more transparent. Formulation and experimentation with the model gives insight into the behaviours. Multi-dimensional rate/level analysis indicates that there is only a narrow region in which the system is able to move downwards. Model runs reproduce the single 'desired' mode of behaviour and all three of the observed 'problematic' ones. Sensitivity analysis gives further insight into the system's modes and their causes. Behaviour is seen to arise only when the geometric effects apply (number of players greater than degrees of freedom of object) in combination with a range of HPB effects. An analogy exists between the co-operative behaviour required here and various examples: conflicting strategic objectives in organizations; Prisoners' Dilemma and integrated bargaining situations. Additionally, the game may be relatable in more direct algebraic terms to situations involving companies in which the resulting behaviours are mediated by market regulations. Finally, comment is offered on the inadequacy of some forms of theory building and the case is made for formal theory building involving the use of models, analysis and plausible explanations to create deep understanding of social phenomena.

Production of attaching-effacing lesions in ligated large intestine loops of 6-month-old sheep by Escherichia coli O157:H7

Shiga-toxigenic Escherichia coli O157:H7 (STEC O157:H7) is associated with potentially fatal human disease, and a persistent reservoir of the organism is present in some farm animal species, especially cattle and sheep. The mechanisms of persistent colonisation of the ruminant intestine by STEC O157:H7 are poorly understood but may be associated with intimate adherence to eukaryotic cells. Intimate adherence, as evidenced by induction of attaching-effacing (AE) lesions by STEC O157, has been observed in 6-day-old conventional lambs after deliberate oral infection but not in older animals. Thus, the present study used a ligated intestinal loop technique to investigate whether STEC O157:H7 and other attaching-effacing E. coli may adhere intimately to the sheep large intestinal mucosa. To do this, four STEC O157:H7 strains, one STEC 026:K60:H11 and one Shiga toxin-negative E. coli O157:H7 strain, suspended in either phosphate-buffered saline or Dulbecco's modified Eagle's medium, were inoculated into ligated spiral colon loops of each of two lambs. The loops were removed 6 h after inoculation, fixed and examined by light and electron microscopy. AE lesions on the intestinal mucosa were produced by all the inoculated strains. However, the lesions were sparse and small, typically comprising bacterial cells intimately adhered to a single enterocyte, or a few adjacent enterocytes. There was little correlation between the extent of intimate adherence in this model and the bacterial cell density, pre-inoculation growth conditions of the bacteria or the strain tested.