Evaluation and comparison of SYBR Green I Real-Time PCR and TaqMan Real-Time PCR methods for quantitative assay of Listeria monocytogenes in nutrient broth and milk

Specific traditional plate count method and real-time PCR systems based on SYBR Green I and TaqMan technologies using a specific primer pair and probe for amplification of iap-gene were used for quantitative assay of Listeria monocytogenes in seven decimal serial dilution series of nutrient broth and milk samples containing 1.58 to 1.58×107 cfu /ml and the real-time PCR methods were compared with the plate count method with respect to accuracy and sensitivity. In this study, the plate count method was performed using surface-plating of 0.1 ml of each sample on Palcam Agar. The lowest detectable level for this method was 1.58×10 cfu/ml for both nutrient broth and milk samples. Using purified DNA as a template for generation of standard curves, as few as four copies of the iap-gene could be detected per reaction with both real-time PCR assays, indicating that they were highly sensitive. When these real-time PCR assays were applied to quantification of L. monocytogenes in decimal serial dilution series of nutrient broth and milk samples, 3.16×10 to 3.16×105 copies per reaction (equals to 1.58×103 to 1.58×107 cfu/ml L. monocytogenes) were detectable. As logarithmic cycles, for Plate Count and both molecular assays, the quantitative results of the detectable steps were similar to the inoculation levels.

Instructions for an Audio Performance/Scissors Music

‘Instructions for an Audio Performance/Scissors Recording’ is a score for a performance using an amplified scissors that can be downloaded and interpreted by a performer anywhere in the world. The file functions as a manual for the performer with guidance for creating the instrument and preparing the performance space while it also provides a template for the actual sonic pattern to be followed in the live performance. Created originally for ‘Storageroom’ an online platform featuring complete exhibitions that are available for download, the piece was exhibited and interpreted in a live performance by Ayelet Lerman for the File Transfer Protocol, the contemporary section of the Haifa-Jerusalem-Tel Aviv exhibition at the Museum of Art, Haifa, Israel.

The development of a ligase mediated PCR with potential for the differentiation of serovars within Leptospira interrogans

A ligase mediated polymerase chain reaction (LMPCR) was developed to amplify between the repetitive element, IS1533, of Leptospira and adjacent chromosomally located BglII restriction endonuclease enzyme sites. To do this, complimentary oligonucleotide linkers designed to anneal together with an overhanging BglII end were ligated to BglII digested DNA from 35 leptospiral reference strains and field isolates, This ligated DNA was used as template for PCR with oligonucleotide primers specific for the linker and for the repetitive element IS1533. The resultant amplicon profile hybridised a 102 hp region derived from the terminus of IS1533 thus confirming that amplicons generated by LMPCR contained part of IS1533. The number of fragments generated containing IS1533 was significantly fewer than that generated by RFLP but the LMPCR method has the potential to use far less template DNA and be quicker than standard RFLP. Obvious and reproducible interserovar differences were demonstrated by LMPCR whereas for 20 of 21 L. hardjo-bovis isolates tested no intraserovar differences were observed. Of those serovars known to possess IS1533 homologues and tested here by LMPCR, each produced a unique amplicon profile which hybridised the IS1533 terminus probe. The limited heterogeneity amongst hardjo-bovis isolates is discussed as is the potential contribution of this method to diagnosis, differentiation and the phylogenetics of the Leptospires.

Venues on the verges: London's theater government between 1594 and 1614

In 1594, major decisions were made by the governors of London and the country about plays and playing. We need to learn what lay behind these events, such as what led James Burbage to build his Blackfriars theater in 1596. That initial fiasco might tell us much about what lay behind Shakespeare’s decision to join the new Chamberlain’s Men in 1594 and his subsequent commitment to them as a full-time playwright. When the Globe burned down in 1613, a majority of the shareholders decided to rebuild it at great cost, but Shakespeare withdrew. The rebuilding was old-fashioned thinking, reverting to the company’s desire, asserted in 1594, to play indoors in winter, which helps to clarify their decisions and Shakespeare’s own—to write plays rather than more long poems. The few surviving papers of the Privy Council and the London mayoralty from the time suggest that one of the two new companies of 1594 preferred to play indoors during the winter instead of at their allocated open playhouses in the suburbs. They tried to renew this traditional practice, first in 1594 and again in 1596 when James Burbage built the indoor Blackfriars playhouse for them. The renewal of the Globe in 1614 was part of the same thinking, although Shakespeare evidently opted out of the decision.

Comparing the thermal performance of horizontal slinky-loop and vertical slinky-loop heat exchangers

The heat pump market in the UK has grown rapidly over the last few years. Performance analyses of vertical ground-loop heat exchanger configurations have been widely carried out using both numerical modelling and experiments. However, research findings and design recommendations on horizontal slinky-loop and vertical slinky-loop heat exchangers are far fewer compared with those for vertical ground-loop heat exchanger configurations, especially where the long-term operation of the systems is concerned. The paper presents the results obtained from a numerical simulation for the horizontal slinky-loop and vertical slinky-loop heat exchangers of a ground-source heat pump system. A three-dimensional numerical heat transfer model was developed to study the thermal performance of various heat exchanger configurations. The influence of the loop pitch (loop spacing) and the depth of a vertical slinky-loop installation were investigated and the thermal performance and excavation work required for the horizontal and vertical slinky-loop heat exchangers were compared. The influence of the installation depth for vertical slinky-loop configurations was also investigated. The results of this study show that the influence of the installation depth of the vertical slinky-loop heat exchanger on the thermal performance of the system is small. The maximum difference in the thermal performance between the vertical and horizontal slinky-loop heat exchangers with the same loop diameter and loop pitch is less than 5%.

Simulation of thermal performance of horizontal slinky-loop heat exchangers for ground source heat pumps

This paper presents results obtained from a numerical simulation for the horizontal slinky-loop heat exchanger of a ground-source heat pump system. A three-dimensional numerical model was developed and the results of the thermal performance of various heat exchanger configurations are presented. The investigation was carried out on five types of loop pitch (loop spacing), three types of loop diameter, three values of soil thermal properties, and allowing continuous and intermittent operation. Comparison was made for the heat transfer rate, the amount of pipe material needed, as well as excavation work required for the horizontal slinky-loop heat exchanger. The results indicate that system parameters have a significant effect on the thermal performance of the system

Preparation of films of a highly aligned lipid cubic phase

We demonstrate a method by which we can produce an oriented film of an inverse bicontinuous cubic phase (QII D) formed by the lipid monoolein (MO). By starting with the lipid as a disordered precursor (the L3 phase) in the presence of butanediol, we can obtain a film of the QII D phase showing a high degree of in-plane orientation by controlled dilution of the sample under shear within a linear flow cell. We demonstrate that the direction of orientation of the film is different from that found in the oriented bulk material that we have reported previously; therefore, we can now reproducibly form QII D samples oriented with either the [110] or the [100] axis aligned in the flow direction depending on the method of preparation. The deposition of MO as a film, via a moving fluid− air interface that leaves a coating of MO in the L3 phase on the capillary wall, leads to a sample in the [110] orientation. This contrasts with the bulk material that we have previously demonstrated to be oriented in the [100] direction, arising from flow producing an oriented bulk slug of material within the capillary tube. The bulk sample contains significant amounts of residual butanediol, which can be estimated from the lattice parameter of the QII D phase obtained. The sample orientation and lattice parameters are determined from synchrotron small-angle X-ray scattering patterns and confirmed by simulations. This has potential applications in the production of template materials and the growth of protein crystals for crystallography as well as deepening our understanding of the mechanisms underlying the behavior of lyotropic liquid-crystal phases.

Location, control and innovation in knowledge-intensive industries

The rising share of intangibles in economies worldwide highlights the crucial role of knowledge-intensive and creative industries in current and future wealth generation. The recognition of this trend has led to intense competition in these industries. At the micro-level, firms from both advanced and emerging economies are globally dispersing their value chains to control costs and leverage capabilities. The geography of innovation is the outcome of a dynamic process whereby firms from emerging economies strive to catch-up with advanced economy competitors, creating strong pressures for continued innovation. However, two distinct strategies can be discerned with regard to the control of the value chain. A vertical integration strategy emphasizes taking advantage of ‘linkage economies’ whereby controlling multiple value chain activities enhances the efficiency and effectiveness of each one of them. In contrast, a specialization strategy focuses on identifying and controlling the creative heart of the value chain, while outsourcing all other activities. The global mobile handset industry is used as the template to illustrate the theory.

Probiotic administration attenuates myocardial hypertrophy and heart failure following myocardial infarction in the rat

Background—Probiotics are extensively used to promote gastrointestinal health and emerging evidence suggests that their beneficial properties can extend beyond the local environment of the gut. Here, we determined whether oral probiotic administration can alter the progression of post-infarction heart failure. Methods and Results—Rats were subjected to six weeks of sustained coronary artery occlusion and administered the probiotic Lactobacillus rhamnosus GR-1 or placebo in the drinking water ad libitum. Culture and 16s rRNA sequencing showed no evidence of GR-1 colonization or a significant shift in the composition of the cecal microbiome. However, animals administered GR-1 exhibited a significant attenuation of left ventricular hypertrophy based on tissue weight assessment as well as gene expression of atrial natriuretic peptide. Moreover, these animals demonstrated improved hemodynamic parameters reflecting both improved systolic and diastolic left ventricular function. Serial echocardiography revealed significantly improved left ventricular parameters throughout the six week follow-up period including a marked preservation of left ventricular ejection fraction as well as fractional shortening. Beneficial effects of GR-1 were still evident in those animals in which GR-1 was withdrawn at four weeks suggesting persistence of the GR-1 effects following cessation of therapy. Investigation of mechanisms showed a significant increase in the leptin to adiponectin plasma concentration ratio in rats subjected to coronary ligation which was abrogated by GR-1. Metabonomic analysis showed differences between sham control and coronary artery ligated hearts particularly with respect to preservation of myocardial taurine levels. Conclusions—The study suggests that probiotics offer promise as a potential therapy for the attenuation of heart failure.

South Korean MNEs' international HRM approach: hybridization of global standards and local practices

This paper analyses the international Human Resource Management (HRM) approaches of Korean Multinational Enterprises (MNEs). Through a study of nine major Korean MNEs’ approaches to subsidiary-HRM, it is argued that the firms pursue hybridization through a blending of localization and global standardization across detailed elements in five broad HRM practice areas. Local discretion is allowed if not counter to global HRM system requirements and “global best practices” used as the template for global standardization of selected HRM elements. This strategic orientation appears to be part of a deliberate response to the “liabilities of origin” born by firms from non-dominant economies.

Analysis of DNA polymorphism in ancient barley herbarium material: validation of the KASP SNP genotyping platform

The availability of crop specimens archived in herbaria and old seed collections represent valuable resources for the analysis of plant genetic diversity and crop domestication. The ability to extract ancient DNA (aDNA) from such samples has recently allowed molecular genetic investigations to be undertaken in ancient materials. While analyses of aDNA initially focused on the use of markers which occur in multiple copies such as the internal transcribed spacer region (ITS) within ribosomal DNA and those requiring amplification of short DNA regions of variable length such as simple sequence repeats (SSRs), emphasis is now moving towards the genotyping of single nucleotide polymorphisms (SNPs), traditionally undertaken in aDNA by Sanger sequencing. Here, using a panel of barley aDNA samples previously surveyed by Sanger sequencing for putative causative SNPs within the flowering-time gene PPD-H1, we assess the utility of the Kompetitive Allele Specific PCR (KASP) genotyping platform for aDNA analysis. We find KASP to out-perform Sanger sequencing in the genotyping of aDNA samples (78% versus 61% success, respectively), as well as being robust to contamination. The small template size (≥46 bp) and one-step, closed-tube amplification/genotyping process make this platform ideally suited to the genotypic analysis of aDNA, a process which is often hampered by template DNA degradation and sample cross-contamination. Such attributes, as well as its flexibility of use and relatively low cost, make KASP particularly relevant to the genetic analysis of aDNA samples. Furthermore, KASP provides a common platform for the genotyping and analysis of corresponding SNPs in ancient, landrace and modern plant materials. The extended haplotype analysis of PPD-H1 undertaken here (allelic variation at which is thought to be important for the spread of domestication and local adaptation) provides further resolution to the previously identified geographic cline of flowering-time allele distribution, illustrating how KASP can be used to aid genetic analyses of aDNA from plant species. We further demonstrate the utility of KASP by genotyping ten additional genetic markers diagnostic for morphological traits in barley, shedding light on the phenotypic traits, alleles and allele combinations present in these unviable ancient specimens, as well as their geographic distributions.

How far do cultural differences explain the differences between nations? implications for HRM

This conceptual paper argues that national differences in human resource management (HRM) matter and can be explained by cultural and institutional theories. We explore briefly the nature of each set of theories and some of the critiques that have been made of them. We argue that in general much of the differences in HRM between countries are determined by institutional factors, and that management has more influence over the effect of national cultural differences than it has over institutional differences and therefore in most cases the appropriate tests will be tests against institutional differences. Where institutions are less constraining, cultural differences may be the appropriate template.

Self-assembly of a dual functional bioactive peptide amphiphile incorporating both matrix metalloprotease substrate and cell adhesion motifs

We describe a bioactive lipopeptide that combines the capacity to promote the adhesion and subsequent self-detachment of live cells, using template-cell-environment feedback interactions. This self-assembling peptide amphiphile comprises a diene-containing hexadecyl lipid chain (C16e) linked to a matrix metalloprotease-cleavable sequence, Thr-Pro-Gly-Pro-Gln-Gly-Ile-Ala-Gly-Gln, and contiguous with a cell-attachment and signalling motif, Arg-Gly-Asp-Ser. Biophysical characterisation revealed that the PA self-assembles into 3 nm diameter spherical micelles above a critical aggregation concentration (cac). In addition, when used in solution at 5–150 nM (well below the cac), the PA is capable of forming film coatings that provide a stable surface for human corneal fibroblasts to attach and grow. Furthermore, these coatings were demonstrated to be sensitive to metalloproteases expressed endogenously by the attached cells, and consequently to elicit the controlled detachment of cells without compromising their viability. As such, this material constitutes a novel class of multi-functional coating for both fundamental and clinical applications in tissue engineering.

New self-assembling multifunctional templates for the biofabrication and controlled self-release of cultured tissue

The need to source live human tissues for research and clinical applications has been a major driving force for the development of new biomaterials. Ideally, these should elicit the formation of scaffold-free tissues with native-like structure and composition. In this study, we describe a biologically interactive coating that combines the fabrication and subsequent self-release of live purposeful tissues using template–cell–environment feedback. This smart coating was formed from a self-assembling peptide amphiphile comprising a proteasecleavable sequence contiguous with a cell attachment and signaling motif. This multifunctional material was subsequently used not only to instruct human corneal or skin fibroblasts to adhere and deposit discreet multiple layers of native extracellular matrix but also to govern their own self-directed release from the template solely through the action of endogenous metalloproteases. Tissues recovered through this physiologically relevant process were carrier-free and structurally and phenotypically equivalent to their natural counterparts. This technology contributes to a new paradigm in regenerative medicine, whereby materials are able to actively direct and respond to cell behavior. The novel application of such materials as a coating capable of directing the formation and detachment of complex tissues solely under physiological conditions can have broad use for fundamental research and in future cell and tissue therapies.

An investigation of the role of business analysts in IS development

Business analysis has developed since the early 1990s as an IS discipline that is concerned with understanding business problems, defining requirements and evaluating relevant solutions. However, this discipline has had limited recognition within the academic community and little research has been conducted into the practices and standards employed by business analysts. This paper reports on a study into business analysis that considered the activities conducted and the outcomes experienced on IS projects. Senior business analysts were interviewed in order to gain insights into the business analyst role and the techniques and approaches applied when conducting this work. The Context, Content, Process, Outcomes framework was adopted as a basis for developing the interview questions. The data collected was analysed using the template analysis technique and the template was based upon this framework. Additional themes concerning aspects of business analysis that may contribute to IS success emerged during data analysis. These included the key business analysis activities and the skills business analysts require to perform these activities. The organisational attitude was also identified as a key factor in enabling the use and contribution of business analysis.