Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield:the use of a respiratory strain as a microbial cell factory

BACKGROUND: Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. RESULTS: Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. CONCLUSIONS: The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass properties do lead to increased volumetric yields without the need to resort to complex control or cultivation schemes. This is anticipated to be of particular value in the production of challenging targets such as membrane proteins.

Medicines non-use in primary care

This study expands the current knowledge base on the nature, causes and fate of unused medicines in primary care. Three methodologies were used and participants for each element were sampled from the population of Eastern Birmingham PCT. A detailed assessment was made of medicines returned to pharmacies and GP surgeries for destruction and a postal questionnaire covering medicines use and disposal was used to patients randomly selected from the electoral roll. The content of this questionnaire was informed by qualitative data from a group interview on the subject. By use of these three methods it was possible to triangulate the data, providing a comprehensive assessment of unused medicines. Unused medicines were found to be ubiquitous in primary care and cardiovascular, diabetic and respiratory medicines are unused in substantial quantities, accounting for a considerable proportion of the total financial value of all unused medicines. Additionally, analgesic and psychoactive medicines were highlighted as being unused in sufficient quantities for concern. Anti-infective medicines also appear to be present and unused in a substantial proportion of patients’ homes. Changes to prescribed therapy and non-compliance were identified as important factors leading to the generation of unused medicines. However, a wide array of other elements influence the quantities and types of medicines that are unused including the concordancy of GP consultations and medication reviews and patient factors such as age, sex or ethnicity. Medicines were appropriately discarded by 1 in 3 patients through return to a medical or pharmaceutical establishment. Inappropriate disposal was by placing in household refuse or through grey and black water with the possibility of hoarding or diversion also being identified. No correlations wre found between the weight of unused medicines and any clinical or financial factor. The study has highlighted unused medicines to be an issue of some concern and one that requires further study.

The value of proprietary health and safety software to proactive health and safety management

There has been little research in health and safety management concernmg the application of information technology to the field. This thesis attempts to stimulate interest in this area by analysing the value of proprietary health and safety software to proactive health and safety management. The thesis is based upon the detailed software evaluation of seven pieces of proprietary health and safety software. It features a discussion concerning the development of information technology and health and safety management, a review of the key issues identified during the software evaluations, an analysis of the commercial market for this type of software, and a consideration of the broader issues which surround the use of this software. It also includes practical guidance for the evaluation, selection, implementation and maintenance of all health and safety management software. This includes a comprehensive software evaluation chart. The implications of the research are considered for proprietary health and safety software, the application of information technology to health and safety management, and for future research.

Simulating the value of collaboration in multi-actor conservation planning

The loss of habitat and biodiversity worldwide has led to considerable resources being spent for conservation purposes on actions such as the acquisition and management of land, the rehabilitation of degraded habitats, and the purchase of easements from private landowners. Prioritising these actions is challenging due to the complexity of the problem and because there can be multiple actors undertaking conservation actions, often with divergent or partially overlapping objectives. We use a modelling framework to explore this issue with a study involving two agents sequentially purchasing land for conservation. We apply our model to simulated data using distributions taken from real data to simulate the cost of patches and the rarity and co-occurence of species. In our model each agent attempted to implement a conservation network that met its target for the minimum cost using the conservation planning software Marxan. We examine three scenarios where the conservation targets of the agents differ. The first scenario (called NGO-NGO) models the situation where two NGOs are both are targeting different sets of threatened species. The second and third scenarios (called NGO-Gov and Gov-NGO, respectively) represent a case where a government agency attempts to implement a complementary conservation network representing all species, while an NGO is focused on achieving additional protection for the most endangered species. For each of these scenarios we examined three types of interactions between agents: i) acting in isolation where the agents are attempting to achieve their targets solely though their own actions ii) sharing information where each agent is aware of the species representation achieved within the other agent’s conservation network and, iii) pooling resources where agents combine their resources and undertake conservation actions as a single entity. The latter two interactions represent different types of collaborations and in each scenario we determine the cost savings from sharing information or pooling resources. In each case we examined the utility of these interactions from the viewpoint of the combined conservation network resulting from both agents' actions, as well as from each agent’s individual perspective. The costs for each agent to achieve their objectives varied depending on the order in which the agents acted, the type of interaction between agents, and the specific goals of each agent. There were significant cost savings from increased collaboration via sharing information in the NGO-NGO scenario were the agent’s representation goals were mutually exclusive (in terms of specie targeted). In the NGO-Gov and Gov-NGO scenarios, collaboration generated much smaller savings. If the two agents collaborate by pooling resources there are multiple ways the total cost could be shared between both agents. For each scenario we investigate the costs and benefits for all possible cost sharing proportions. We find that there are a range of cost sharing proportions where both agents can benefit in the NGO-NGO scenarios while the NGO-Gov and Gov-NGO scenarios again showed little benefit. Although the model presented here has a range of simplifying assumptions, it demonstrates that the value of collaboration can vary significantly in different situations. In most cases, collaborating would have associated costs and these costs need to be weighed against the potential benefits from collaboration. The model demonstrates a method for determining the range of collaboration costs that would result in collaboration providing an efficient use of scarce conservation resources.

Characterisation of waste derived intermediate pyrolysis oils for use as diesel engine fuels

This paper studies the characteristics of intermediate pyrolysis oils derived from sewage sludge and de-inking sludge (a paper industry residue), with a view to their use as fuels in a diesel engine. The feedstocks were dried and pelletised, then pyrolysed in the Pyroformer intermediate pyrolysis system. The organic fraction of the oils was separated from the aqueous phase and characterised. This included elemental and compositional analysis, heating value, cetane index, density, viscosity, surface tension, flash point, total acid number, lubricity, copper corrosion, water, carbon residue and ash content. Most of these results are compared with commercial diesel and biodiesel. Both pyrolysis oils have high carbon and hydrogen contents and their higher heating values compare well with biodiesel. The water content of the pyrolysis oils is reasonable and the flash point is found to be high. Both pyrolysis oils have good lubricity, but show some corrosiveness. Cetane index is reduced, which may influence ignition. Also viscosity is increased, which may influence atomisation quality. Carbon residue and ash content are both high, indicating potential deposition problems. Compared with de-inking sludge pyrolysis oil (DSPO), sewage sludge pyrolysis oil (SSPO) has a higher heating value, but higher corrosiveness and viscosity. The conclusions are that both intermediate pyrolysis oils will be able to provide sufficient heat when used in diesel engine; however poor combustion and carbon deposition may be encountered. Blending of these pyrolysis oils with diesel or biodiesel could overcome these problems and is recommended for further investigation.

Simulating the value of collaboration in multi-actor conservation planning

The loss of habitat and biodiversity worldwide has led to considerable resources being spent on conservation interventions. Prioritising these actions is challenging due to the complexity of the problem and because there can be multiple actors undertaking conservation actions, often with divergent or partially overlapping objectives. We explore this issue with a simulation study involving two agents sequentially purchasing land for the conservation of multiple species using three scenarios comprising either divergent or partially overlapping objectives between the agents. The first scenario investigates the situation where both agents are targeting different sets of threatened species. The second and third scenarios represent a case where a government agency attempts to implement a complementary conservation network representing 200 species, while a non-government organisation is focused on achieving additional protection for the ten rarest species. Simulated input data was generated using distributions taken from real data to model the cost of parcels, and the rarity and co-occurrence of species. We investigated three types of collaborative interactions between agents: acting in isolation, sharing information and pooling resources with the third option resulting in the agents combining their resources and effectively acting as a single entity. In each scenario we determine the cost savings when an agent moves from acting in isolation to either sharing information or pooling resources with the other agent. The model demonstrates how the value of collaboration can vary significantly in different situations. In most cases, collaborating would have associated costs and these costs need to be weighed against the potential benefits from collaboration. Our model demonstrates a method for determining the range of costs that would result in collaboration providing an efficient use of scarce conservation resources.

The innovation value chain in new technology-based firms:evidence from the U.K.

Innovation is central to the survival and growth of firms, and ultimately to the health of the economies of which they are part. A clear understanding both of the processes by which firms perform innovation and the benefits which flow from innovation in terms of productivity and growth is therefore essential. This paper demonstrates the use of a conceptual framework and modeling tool, the innovation value chain (IVC), and shows how the IVC approach helps to highlight strengths and weaknesses in the innovation performance of a key group of firms-new technology-based firms. The value of the IVC is demonstrated in showing the key interrelationships in the whole process of innovation from sourcing knowledge through product and process innovation to performance in terms of the growth and productivity outcomes of different types of innovation. The use of the IVC highlights key complementarities, such as that between internal R&D, external R&D, and other external sources of knowledge. Other important relationships are also highlighted. Skill resources matter throughout the IVC, being positively associated with external knowledge linkages and innovation success, and also having a direct influence on growth independent of the effect on innovation. A key benefit of the IVC approach is therefore its ability to highlight the roles of different factors at various stages of the knowledge-innovation-performance nexus, and to show their indirect as well as direct impact. This in turn permits both managerial and policy implications to be drawn. © 2012 Product Development & Management Association.

An integral equation method for a mixed initial boundary value problem for unsteady Stokes system in a doubly-connected domain

We present a novel numerical method for a mixed initial boundary value problem for the unsteady Stokes system in a planar doubly-connected domain. Using a Laguerre transformation the unsteady problem is reduced to a system of boundary value problems for the Stokes resolvent equations. Employing a modied potential approach we obtain a system of boundary integral equations with various singularities and we use a trigonometric quadrature method for their numerical solution. Numerical examples are presented showing that accurate approximations can be obtained with low computational cost.

A novel dried blood spot-LCMS method for the quantification of methotrexate polyglutamates as a potential marker for methotrexate use in children

Objective: Development and validation of a selective and sensitive LCMS method for the determination of methotrexate polyglutamates in dried blood spots (DBS). Methods: DBS samples [spiked or patient samples] were prepared by applying blood to Guthrie cards which was then dried at room temperature. The method utilised 6-mm disks punched from the DBS samples (equivalent to approximately 12 μl of whole blood). The simple treatment procedure was based on protein precipitation using perchloric acid followed by solid phase extraction using MAX cartridges. The extracted sample was chromatographed using a reversed phase system involving an Atlantis T3-C18 column (3 μm, 2.1x150 mm) preceded by Atlantis guard column of matching chemistry. Analytes were subjected to LCMS analysis using positive electrospray ionization. Key Results: The method was linear over the range 5-400 nmol/L. The limits of detection and quantification were 1.6 and 5 nmol/L for individual polyglutamates and 1.5 and 4.5 nmol/L for total polyglutamates, respectively. The method has been applied successfully to the determination of DBS finger-prick samples from 47 paediatric patients and results confirmed with concentrations measured in matched RBC samples using conventional HPLC-UV technique. Conclusions and Clinical Relevance: The methodology has a potential for application in a range of clinical studies (e.g. pharmacokinetic evaluations or medication adherence assessment) since it is minimally invasive and easy to perform, potentially allowing parents to take blood samples at home. The feasibility of using DBS sampling can be of major value for future clinical trials or clinical care in paediatric rheumatology. © 2014 Hawwa et al.

Does the importance of value, brand and relationship equity for customer loyalty differ between Eastern and Western cultures?

The use of the customer equity framework as a focal marketing strategy to increase customer loyalty has emerged as an important topic. Despite a growing number of investigations, previous studies are limited by their strong U.S. and European orientations. Research into Western consumers cannot necessarily predict the behaviour of Eastern consumers though. Therefore, this study investigates whether the link between customer equity drivers (value equity, brand equity and relationship equity) and loyalty intentions is sensitive to the cultural environment. A sample of 1553 Chinese and 1085 Dutch consumers in the banking and supermarket industries reveals that all three customer equity drivers exert a greater impact in Western than in Eastern cultures. This study also shows that Eastern consumers in general have higher loyalty intentions than Western consumers. © 2013.

Understanding high-value manufacturing in Scottish SMEs

Purpose: The purpose of this paper is to investigate the use of high-value manufacturing (HVM) concepts in Scottish SMEs and define how they are being used to gain competitive advantage. Design/methodology/approach: Cross-sectional research carried out using a large-scale survey of 435 SMEs and semi-structured interviews of a subset of 50 SMEs. Findings: Findings indicate that HVM is not a homogeneous state but an umbrella term for a number of operational models adopted by manufacturers that are progressively moving from simple price-based production; companies must, as a foundation, be operationally excellent in all lifecycle phases before extending their capability by offering a more comprehensive service; HVM is not a static state but a journey that differs in nature for each manufacturer depending on the nature of its market and customer. Research limitations/implications: The approach to theory must be more integrated combining aspects of marketing, strategic and operational theory. Research must be carried out using the supply chain, rather than the firm, as the unit of analysis. Practical implications: Manufacturing efficiency has now become an order qualifier and competitive advantage should now be sought through the integration of design, production and service activities from strategic levels down to operational levels across all the functions of a business which link seamlessly to customer and supplier activities. Originality/value: This paper contains insights into Scottish SMEs and their practice of HVM; defines the activity that makes up HVM at an operational as opposed to an economic or strategic level; proposes a model that characterises the stages of HVM that SMEs transition through. © Emerald Group Publishing Limited.

The use of maturity models in improving project management performance:an empirical investigation

Purpose – The purpose of the paper was to conduct an empirical investigation to explore the impact of project management maturity models (PMMMs) on improving project performance. Design/methodology/approach – The investigation used a cross-case analysis involving over 90 individuals in seven organisations. Findings – The findings of the empirical investigation indicate that PMMMs demonstrate very high levels of variability in individual's assessment of project management maturity. Furthermore, at higher levels of maturity, the type of performance improvement adopted following their application is related to the type of PMMM used in the assessment. The paradox of the unreliability of PMMMs and their widespread acceptance is resolved by calling upon the “wisdom of crowds” phenomenon which has implications for the use of maturity model assessments in other arena. Research limitations/implications – The investigation does have the usual issues associated with case research, but the steps that have been taken in the cross-case construction and analysis have improved the overall robustness and extendibility of the findings. Practical implications – The tendency for PMMMs to shape improvements based on their own inherent structure needs further understanding. Originality/value – The use of empirical methods to investigate the link between project maturity models and extant changes in project management performance is highly novel and the findings that result from this have added resonance.

The value of risk reporting:a critical analysis of value-at-risk disclosures in the banking sector

This paper critically reviews the evolution of financial reporting in the banking sector with specific reference to the reporting of market risk and the growing use of the measure known as Value at Risk (VaR). The paper investigates the process by which VaR became 'institutionalised'. The analysis highlights a number of inherent limitations of VaR as a risk measure and questions the usefulness of published VaR disclosures, concluding that risk 'disclosure' might be more apparent than real. It also looks at some of the implications for risk reporting practice and the accounting profession more generally.

What difference doguidelines make? An observational study of online-questionnaire design guidelines put to practical use

As a new medium for questionnaire delivery, the Internet has the potential to revolutionize the survey process. Online-questionnaires can provide many capabilities not found in traditional paper-based questionnaires. Despite this, and the introduction of a plethora of tools to support online-questionnaire creation, current electronic survey design typically replicates the look-and-feel of paper-based questionnaires, thus failing to harness the full power of the electronic delivery medium. A recent environmental scan of online-questionnaire design tools found that little, if any, support is incorporated within these tools to guide questionnaire designers according to best-practice [Lumsden & Morgan 2005]. This paper briefly introduces a comprehensive set of guidelines for the design of online-questionnaires. Drawn from relevant disparate sources, all the guidelines incorporated within the set are proven in their own right; as an initial assessment of the value of the set of guidelines as a practical reference guide, we undertook an informal study to observe the effect of introducing the guidelines into the design process for a complex online-questionnaire. The paper discusses the qualitative findings — which are encouraging for the role of the guidelines in the ‘bigger picture’ of online survey delivery across many domains such as e-government, e-business, and e-health — of this case study.

Polarization effects in polymer FBGs:study and use for transverse force sensing

Bragg gratings photo-inscribed in polymer optical fibers (POFs) are more sensitive to temperature and pressure than their silica counterparts, because of their larger thermo-optic coefficient and smaller Young's modulus. Polymer optical fiber Bragg gratings (POFBGs) are most often photo-written in poly(methylmethacrylate) (PMMA) based materials using a continuous-wave 325 nm HeCd laser. In this work, we present the first study about birefringence effects in POFBGs manufactured in different types of fiber. To achieve this, highly reflective (> 90%) gratings were produced with the phase mask technique. Their spectral response was then monitored in transmission with polarized light. Polarization dependent loss (PDL) and differential group delay (DGD) were computed from the Jones matrix eigenanalysis using an optical vector analyzer. Maximum values exceeding several dB and a few picoseconds were obtained for the PDL and DGD, respectively. An inverse scattering technique applied to the experimental data provided an estimate of the photo-induced birefringence value arising from the side fabrication process. The response to lateral force was finally investigated for various incident directions using the PDL response of FBGs manufactured in step-index POFs. As the force induced birefringence adds to the photo-induced one, a force dependent evolution of the PDL maximum value was noticed, with a good temperature-insensitivity.