SEF17 fimbriae are essential for the convoluted colonial morphology of Salmonella enteritidis

Salmonella enteritidis isolated from poultry infections generated a convoluted colonial morphology after 48 h growth on colonisation factor antigen (CFA) agar at 25 degrees C. A mutant S. enteritidis defective for the elaboration of the SEF17 fimbrial antigen, in which the agf gene cluster was inactivated by insertion of an ampicillin resistance gene cassette, and other wild-type S. enteritidis transduced to this genotype failed to produce convoluted colonies. However, growth of SEF17(-) mutans at 25 degrees C on CFA agar supplemented with 0.001% Congo red resulted in partial recovery of the phenotype. Immunoelectron microscopy demonstrated that copious amounts of the SEF17 fimbrial antigen were present in the extracellular matrix of convoluted colonies of wild-type virulent S. enteritidis isolates. Bacteria were often hyperflagellated also. Immunoelectron microscopy of SEF17(-) mutants grown on CFA agar+0.001% Congo red demonstrated the elaboration of an as yet undefined fimbrial structure. Isolates of S. enteritidis which were described previously as avirulent and sensitive to environmental stress failed to express SEF17 or produce convoluted colonies. These data indicate an essential role for SEF17, and possibly for another fimbria and flagella, in the generation of the convoluted colonial phenotype. The relationship between virulence and colonial phenotype is discussed.

Facilities management carbon footprints: an audit of critical elements of management and reporting

Concern for the environmental impact of organizations’ activities has led to the recognition and demand for organizations to manage and report on their carbon footprint. However, there is no limit as to the areas of carbon footprints required in such annual environmental reports. To deliver improvements in the quality of carbon footprint management and reporting, there is a need to identify the main elements of carbon footprint strategy that can be endorsed, supported and encouraged by facility managers. The study investigates carbon footprint elements managed and reported upon by facility manager in the UK. Drawing on a questionnaire survey of 256 facility managers in the UK, the key elements of carbon footprints identified in carbon footprint reports are examined. The findings indicate that the main elements are building energy consumption, waste disposal and water consumption. Business travel in terms of using public transport, air travel and company cars are also recognized as important targets and objectives for the carbon footprint strategy of several FM (facilities management) organizations.

Platelet protein disulfide isomerase is required for thrombus formation but not essential for hemostasis in mice

Protein disulfide isomerase (PDI) derived from intravascular cells is required for thrombus formation. However, it remains unclear whether platelet PDI contributes to the process. Using platelet-specific PDI-deficient mice, we demonstrate that PDI-null platelets have defects in aggregation and ATP secretion induced by thrombin, collagen, and ADP. Such defects were rescued by exogenously-added wild-type but not mutant PDI, indicating that the isomerase activity of platelet surface PDI is critical for the regulatory effect. PDI-deficient platelets expressed increased levels of intracellular ERp57 and ERp72. Platelet PDI regulated αIIbβ3 integrin activation but not P-selectin exposure, Ca2+ mobilization, β3-talin interaction, and platelet spreading on immobilized fibrinogen. Inhibition of ERp57 further diminished αIIbβ3 integrin activation, aggregation and ATP secretion of activated PDI-deficient platelets, suggesting distinct roles of PDI and ERp57 in platelet functions. We found that platelet PDI is important for thrombus formation on collagen-coated surfaces under arteriolar shear. Intravital microscopy demonstrates that platelet PDI is important for platelet accumulation but not initial adhesion and fibrin generation following laser-induced arteriolar injury. Tail bleeding time and blood loss in platelet-specific PDI-deficient mice were not significantly increased. Our results provide important evidence that platelet PDI is essential for thrombus formation but not for hemostasis in mice.

The impact of eBusiness on the City of London office market

The City of London is a major financial centre where the property market is heavily dependent on the space requirements of office occupiers. Technological change, however, is changing the way in which organisations conduct their business and is challenging the continued locational pull of the City. This research presents findings from a major survey of City occupiers carried out by The College research team during 2002. The research examines how the shape and form of the office property market in the City will be affected by process change in eBusiness and related technology over the next 5 years. The report, which includes an extensive literature review, examines the implications of increased homeworking and other trends for future office supply in the City. The research was funded by the Worshipful Company of Chartered Surveyors Trust (Toby Sutton Research Award), and part of the literature review by the Small Business Support Research Unit of DTI. It is essential reading for surveyors, planners, facilities managers and all those interested in the technology and property interface.

The ESA climate change initiative: satellite data records for essential climate variables

Observations of Earth from space have been made for over 40 years and have contributed to advances in many aspects of climate science. However, attempts to exploit this wealth of data are often hampered by a lack of homogeneity and continuity and by insufficient understanding of the products and their uncertainties. There is, therefore, a need to reassess and reprocess satellite datasets to maximize their usefulness for climate science. The European Space Agency has responded to this need by establishing the Climate Change Initiative (CCI). The CCI will create new climate data records for (currently) 13 essential climate variables (ECVs) and make these open and easily accessible to all. Each ECV project works closely with users to produce time series from the available satellite observations relevant to users' needs. A climate modeling users' group provides a climate system perspective and a forum to bring the data and modeling communities together. This paper presents the CCI program. It outlines its benefit and presents approaches and challenges for each ECV project, covering clouds, aerosols, ozone, greenhouse gases, sea surface temperature, ocean color, sea level, sea ice, land cover, fire, glaciers, soil moisture, and ice sheets. It also discusses how the CCI approach may contribute to defining and shaping future developments in Earth observation for climate science.

The dermomyotome ventrolateral lip is essential for the hypaxial myotome formation

Background The myotome is the primitive skeletal muscle that forms within the embryonic metameric body wall. It can be subdivided into an epaxial and hypaxial domain. It has been shown that the formation of the epaxial myotome requires the dorsomedial lip of the dermomyotome (DML). Although the ventrolateral lip (VLL) of the dermomyotome is believed to be required for the formation of the hypaxial myotome, experimentally evidence for this statement still needs to be provided. Provision of such data would enable the resolution of a debate regarding the formation of the hypaxial dermomyotome. Two mechanisms have been proposed for this tissue. The first proposes that the intermediate dermomyotome undergoes cellular expansion thereby pushing the ventral lateral lip in a lateral direction (translocation). In contrast, the alternative view holds that the ventral lateral lip grows laterally. Results Using time lapse confocal microscopy, we observed that the GFP-labelled ventrolateral lip (VLL) of the dermomyotome grows rather than translocates in a lateral direction. The necessity of the VLL for lateral extension of the myotome was addressed by ablation studies. We found that the hypaxial myotome did not form after VLL ablation. In contrast, the removal of an intermediate portion of the dermomyotome had very little effect of the hypaxial myotome. These results demonstrate that the VLL is required for the formation of the hypaxial myotome. Conclusion Our study demonstrates that the dermomyotome ventrolateral lip is essential for the hypaxial myotome formation and supports the lip extension model. Therefore, despite being under independent signalling controls, both the dorsomedial and ventrolateral lip fulfil the same function, i.e. they extend into adjacent regions permitting the growth of the myotome.

Integrin-linked kinase regulates the rate of platelet activation and is essential for the formation of stable thrombi

BACKGROUND: Integrin-linked kinase (ILK) and its associated complex of proteins are involved in many cellular activation processes, including cell adhesion and integrin signaling. We have previously demonstrated that mice with induced platelet ILK deficiency show reduced platelet activation and aggregation, but only a minor bleeding defect. Here, we explore this apparent disparity between the cellular and hemostatic phenotypes. METHODS: The impact of ILK inhibition on integrin αII b β3 activation and degranulation was assessed with the ILK-specific inhibitor QLT0267, and a conditional ILK-deficient mouse model was used to assess the impact of ILK deficiency on in vivo platelet aggregation and thrombus formation. RESULTS: Inhibition of ILK reduced the rate of both fibrinogen binding and α-granule secretion, but was accompanied by only a moderate reduction in the maximum extent of platelet activation or aggregation in vitro. The reduction in the rate of fibrinogen binding occurred prior to degranulation or translocation of αII b β3 to the platelet surface. The change in the rate of platelet activation in the absence of functional ILK led to a reduction in platelet aggregation in vivo, but did not change the size of thrombi formed following laser injury of the cremaster arteriole wall in ILK-deficient mice. It did, however, result in a marked decrease in the stability of thrombi formed in ILK-deficient mice. CONCLUSION: Taken together, the findings of this study indicate that, although ILK is not essential for platelet activation, it plays a critical role in facilitating rapid platelet activation, which is essential for stable thrombus formation.

Determination of the absolute accuracy of UK chamber facilities used in measuring methane emissions from livestock

Respiration chambers are one of the primary sources of data on methane emissions from livestock. This paper describes the results from a coordinated set of chamber validation experiments which establishes the absolute accuracy of the methane emission rates measured by the chambers, and for the first time provides metrological traceability to international standards, assesses the impact of both analyser and chamber response times on measurement uncertainty and establishes direct comparability between measurements made across different facilities with a wide range of chamber designs. As a result of the validation exercise the estimated combined uncertainty associated with the overall capability across all facilities reduced from 25.7% (k = 2, 95% confidence) before the validation to 2.1% (k = 2, 95% confidence) when the validation results are applied to the facilities’ data.

What is outside? The early years foundation stage in England: outdoor facilities, organisation and staff attitudes

The purpose of this paper is to report on the facilities available, organisation of, and staff attitudes to early years outdoor education from schools within the south east of England, focusing on provision for children aged three to five. One component of the successful education of the child involves providing an ‘environment for learning’, including the facilities, layout and routines. This paper presents findings concerning the type and variety of facilities available outside; the various styles of organisation of the space; staff attitudes about: their roles, their aims for the environment, children’s behaviour and learning, and perceived drawbacks to practice. This paper draws on empirical data collected from schools within the University of Reading partnership. The findings suggest that although all early years settings must adhere to the statutory framework there are a range of facilities available, and there are a number of ways this environment is organised. Further there appears to be uncertainty about the adult role outside and the aims for activities. The conclusions drawn indicate that staff do not appear to be linking their aims for outdoor education to the facilities provided or to their actions outside. This means there is not a clear link between what staff provide outside and the declared ambitions for learning. This study is important as all educators need to be certain about their aims for education to ensure best outcomes for children. The implications of these findings for early years teachers are that they need to be able to articulate their aims for outdoor education and to provide the correct facilities to achieve these aims. Finally this study was undertaken to raise debate, posit questions and to ascertain the parameters for further research about the early years outdoor environment.

Nicked-site substrates for a serine recombinase reveal enzyme–DNA communications and an essential tethering role of covalent enzyme–DNA linkages

To analyse the mechanism and kinetics of DNA strand cleavages catalysed by the serine recombinase Tn3 resolvase, we made modified recombination sites with a single-strand nick in one of the two DNA strands. Resolvase acting on these sites cleaves the intact strand very rapidly, giving an abnormal half-site product which accumulates. We propose that these reactions mimic second-strand cleavage of an unmodified site. Cleavage occurs in a synapse of two sites, held together by a resolvase tetramer; cleavage at one site stimulates cleavage at the partner site. After cleavage of a nicked-site substrate, the half-site that is not covalently linked to a resolvase subunit dissociates rapidly from the synapse, destabilizing the entire complex. The covalent resolvase–DNA linkages in the natural reaction intermediate thus perform an essential DNA-tethering function. Chemical modifications of a nicked-site substrate at the positions of the scissile phosphodiesters result in abolition or inhibition of resolvase-mediated cleavage and effects on resolvase binding and synapsis, providing insight into the serine recombinase catalytic mechanism and how resolvase interacts with the substrate DNA.

Developing temporary manufacturing facilities for residential building: a case of the modern flying factory

The building industry is often berated for its short comings in meeting up with the demand for the provision of new housing. Addressing the need for new housing stock is a challenge that has led to debates among professional bodies, the construction sector, housing industry and government. The introduction of new manufacturing technologies is often offered as a solution, but the challenges of increasing the amount of off-site construction in residential building are well known and well-rehearsed. The modern flying factory (MFF) is a concept that involves the manufacture of specific components or modules in temporary off- or near- site locations using relatively simple and quick to set up and dismantle technologies and processes. The aim is to produce short batches and hence achieve some of the benefits of off-site manufacture on a much smaller scale than in dedicated factory environments. A case study of a modern flying factory being set up to produce pre-assembled utility cupboards for a large residential development in London is presented, involving participant observation and informal interviews with key actors on the design and operationalising of the process. The case reveals that although there are costs, efficiency and health and safety benefits to using MFF approaches, there are also challenges to overcome over the time required to set up and establish the process for relatively short runs, and in evaluating whether the MFF or traditional site based production is most effective for particular aspects of projects.

CLEC-2 and Syk in the megakaryocytic/platelet lineage are essential for development

The C-type lectin receptor CLEC-2 signals through a pathway that is critically dependent on the tyrosine kinase Syk. We show that homozygous loss of either protein results in defects in brain vascular and lymphatic development, lung inflation, and perinatal lethality. Furthermore, we find that conditional deletion of Syk in the hematopoietic lineage, or conditional deletion of CLEC-2 or Syk in the megakaryocyte/platelet lineage, also causes defects in brain vascular and lymphatic development, although the mice are viable. In contrast, conditional deletion of Syk in other hematopoietic lineages had no effect on viability or brain vasculature and lymphatic development. We show that platelets, but not platelet releasate, modulate the migration and intercellular adhesion of lymphatic endothelial cells through a pathway that depends on CLEC-2 and Syk. These studies found that megakaryocyte/platelet expression of CLEC-2 and Syk is required for normal brain vasculature and lymphatic development and that platelet CLEC-2 and Syk directly modulate lymphatic endothelial cell behavior in vitro.