Characterisation of a harvesting step for monoclonal antibodies from mammalian cell culture

The focus of this research was defined by a poorly characterised filtration train employed to clarify culture broth containing monoclonal antibodies secreted by GS-NSO cells: the filtration train blinded unpredictably and the ability of the positively charged filters to adsorb DNA from process material was unknown. To direct the development of an assay to quantify the ability of depth filters to adsorb DNA, the molecular weight of DNA from a large-scale, fed-batch, mammalian cell culture vessel was evaluated as process material passed through the initial stages of the purification scheme. High molecular weight DNA was substantially cleared from the broth after passage through a disc stack centrifuge and the remaining low molecular weight DNA was largely unaffected by passage through a series of depth filters and a sterilising grade membrane. Removal of high molecular weight DNA was shown to be coupled with clarification of the process stream. The DNA from cell culture supernatant showed a pattern of internucleosomal cleavage of chromatin when fractionated by electrophoresis but the presence of both necrotic and apoptotic cells throughout the fermentation meant that the origin of the fragmented DNA could not be unequivocally determined. An intercalating fluorochrome, PicoGreen, was elected for development of a suitable DNA assay because of its ability to respond to low molecular weight DNA. It was assessed for its ability to determine the concentration of DNA in clarified mammalian cell culture broths containing pertinent monoclonal antibodies. Fluorescent signal suppression was ameliorated by sample dilution or by performing the assay above the pI of secreted IgG. The source of fluorescence in clarified culture broth was validated by incubation with RNase A and DNase I. At least 89.0 % of fluorescence was attributable to nucleic acid and pre-digestion with RNase A was shown to be a requirement for successful quantification of DNA in such samples. Application of the fluorescence based assay resulted in characterisation of the physical parameters governing adsorption of DNA by various positively charged depth filters and membranes in test solutions and the DNA adsorption profile of the manufacturing scale filtration train. Buffers that reduced or neutralised the depth filter or membrane charge, and those that impeded hydrophobic interactions were shown to affect their operational capacity, demonstrating that DNA was adsorbed by a combination of electrostatic and hydrophobic interactions. Production-scale centrifugation of harvest broth containing therapeutic protein resulted in the reduction of total DNA in the process stream from 79.8 μg m1-1 to 9.3 μg m1-1 whereas the concentration of DNA in the supernatant of pre-and post-filtration samples had only marginally reduced DNA content: from 6.3 to 6.0 μg m1-1 respectively. Hence the filtration train was shown to ineffective in DNA removal. Historically, blinding of the depth filters had been unpredictable with data such as numbers of viable cells, non-viable cells, product titre, or process shape (batch, fed-batch, or draw and fill) failing to inform on the durability of depth filters in the harvest step. To investigate this, key fouling contaminants were identified by challenging depth filters with the same mass of one of the following: viable healthy cells, cells that had died by the process of apoptosis, and cells that had died through the process of necrosis. The pressure increase across a Cuno Zeta Plus 10SP depth filter was 2.8 and 16.5 times more sensitive to debris from apoptotic and necrotic cells respectively, when compared to viable cells. The condition of DNA released into the culture broth was assessed. Necrotic cells released predominantly high molecular weight DNA in contrast to apoptotic cells which released chiefly low molecular weight DNA. The blinding of the filters was found to be largely unaffected by variations in the particle size distribution of material in, and viscosity of, solutions with which they were challenged. The exceptional response of the depth filters to necrotic cells may suggest the cause of previously noted unpredictable filter blinding whereby a number of necrotic cells have a more significant impact on the life of a depth filter than a similar number of viable or apoptotic cells. In a final set of experiments the pressure drop caused by non-viable necrotic culture broths which had been treated with DNase I or benzonase was found to be smaller when compared to untreated broths: the abilities of the enzyme treated cultures to foul the depth filter were reduced by 70.4% and 75.4% respectively indicating the importance of DNA in the blinding of the depth filter studied.

A survey on the adoption and usage of broadband internet

In spite of the increasing significance of broadband Internet, there are not many research papers explicitly addressing issues pertaining to its adoption and postadoption. Previous research on broadband has mainly focused on the supply side aspect at the national level, ignoring the importance of the demand side which may involve looking more deeply into the use, as well as factors impacting organizational and individual uptake. In an attempt to fill this gap, the current study empirically verifies an integrated theoretical model comprising the theory of planned behavior and the IS continuance model to examine factors influencing broadband Internet adoption and postadoption behavior of some 1,500 organizations in Singapore. Overall, strong support for the integrated model has been manifested by our results, providing insight into influential factors. At the adoption stage, perceived behavioral control has the greatest impact on behavioral intention. Our findings also suggest that, as compared to attitude, subjective norms and perceived behavioral control more significantly affect the broadband Internet adoption decision. At the postadoption stage, intention is no longer the only determinant of broadband Internet continuance; rather, initial usage was found to significantly affect broadband Internet continuance.

Harnessing diversity and collective action in the top management team

Diversity has become an important issue at all levels of the company from the boardroom to the back office. It is increasingly apparent that diversity is vital to productivity, with academic research indicating an important link between diverse top management team (TMT) composition and corporate performance. However, the nature of this link remains elusive, as there is little accessible research that can help top teams to evaluate how diversity impacts on their strategic capacity. This paper seeks to fill this gap by developing a conceptual framework, illustrated with case examples, to explain the relationships between TMT diversity and TMT collective action. As collective action is difficult to attain from top teams that are high in diversity, six practical processes are developed from this framework for establishing and exploiting top team strategic capacity. The paper concludes by outlining the theoretical implications of the framework. © Elsevier Ltd. All rights reserved.

Advances in femtosecond micromachining and inscription of micro and nano photonic devices

This thesis has focused on three key areas of interest for femtosecond micromachining and inscription. The first area is micromachining where the work has focused on the ability to process highly repeatable, high precision machining with often extremely complex geometrical structures with little or no damage. High aspect ratio features have been demonstrated in transparent materials, metals and ceramics. Etch depth control was demonstrated especially in the work on phase mask fabrication. Practical chemical sensing and microfluidic devices were also fabricated to demonstrate the capability of the techniques developed during this work. The second area is femtosecond inscription. Here, the work has utilised the non-linear absorption mechanisms associated with femtosecond pulse-material interactions to create highly localised refractive index changes in transparent materials to create complex 3D structures. The techniques employed were then utilised in the fabrication of Phase masks and Optical Coherence Tomography (OCT) phantom calibration artefacts both of which show the potential to fill voids in the development of the fields. This especially the case for the OCT phantoms where there exists no previous artefacts of known shape, allowing for the initial specification of parameters associated with the quality of OCT machines that are being taken up across the world in industry and research. Finally the third area of focus was the combination of all of the techniques developed through work in planar samples to create a range of artefacts in optical fibres. The development of techniques and methods for compensating for the geometrical complexities associated with working with the cylindrical samples with varying refractive indices allowed for fundamental inscription parameters to be examined, structures for use as power monitors and polarisers with the optical fibres and finally the combination of femtosecond inscription and ablation techniques to create a magnetic field sensor with an optical fibre coated in Terfenol-D with directional capability. Through the development of understanding, practical techniques and equipment the work presented here demonstrates several novel pieces of research in the field of femtosecond micromachining and inscription that has provided a broad range of related fields with practical devices that were previously unavailable or that would take great cost and time to facilitate.

Institutions and firm ownership structure:evidence from Central and Eastern Europe

FDI plays a key role in development, particularly in resource-constrained transition economies of Central and Eastern Europe with relatively low savings rates. Gains from technology transfer play a critical role in motivating FDI, yet potential for it may be hampered by a large technology gap between the source and host country. While the extent of this gap has traditionally been attributed to education, skills and capital intensity, recent literature has also emphasized the possible role of institutional environment in this respect. Despite tremendous interest among policy-makers and academics to understand the factors attracting FDI (Bevan and Estrin, 2000; Globerman and Shapiro, 2003) our knowledge about the effects of institutions on the location choice and ownership structure of foreign firms remains limited. This paper attempts to fill this gap in the literature by examining the link between institutions and foreign ownership structures. To the best of our knowledge, Javorcik (2004) is the only papers, which use firm-level data to analyse the role of institutional quality on an outward investor’s entry mode in transition countries. Our paper extends Javorcik (2004) in a number of ways: (a) rather than a cross-section, we use panel data for the period 1997-2006; (b) rather than a binary variable, we use the percentage foreign ownership as continuous variable; (c) we consider multi-dimensional institutional variables, such as corruption, intellectual property rights protection and government stability. We also use factor analysis to generate a composite index of institutional quality and see how stronger institutional environment could affect foreign ownership; (d) we explore how the distance between institutional environment in source and host countries affect foreign ownership in a host country. The firm-level data used includes both domestic and foreign firms for the period 1997-2006 and is drawn from ORBIS, a commercially available dataset provided by Bureau van Dijk. In order to examine the link between institutions and foreign ownership structures, we estimate four log-linear ownership equations/specifications augmented by institutional and other control variables. We find evidence that the decision of a foreign firm to either locate its subsidiary or acquire an existing domestic firm depends not only on factor cost differences but also on differences in institutional environment between the host and source countries.

Determinant of broadband internet adoption and continuance in Singapore

In spite of increasing significance of Broadband Internet, there are not many research papers explicitly addressing issues pertaining to its deployment and continuance. Previous research on Broadband has mainly focused on the supply side aspect at the national level, ignoring the importance of the demand side which may involve looking more deeply into the factors impacting organizational and individual uptake. In an attempt to fill this gap, the current study empirically verifies the IS continuance model to examine factors influencing Broadband Internet post-adoption behavior of some 1,500 organizations in Singapore. Strong support for the model has been manifested by our results, providing insight into influential factors. Results of the study suggest that that perceived usefulness is the strongest predictor of users' continuance intention, followed by satisfaction with Broadband Internet usage as a significant but weaker predictor.

Ecological studies on high-rate biological filters with special reference to microbial biosynthesis and nitrification

Pilot scale studies of high rate filtration were initiated to assess its potential as either a primary 'roughing' filter to alleviate the seasonal overloading of low rate filters on Hereford sewage treatment works - caused by wastes from cider production - or as a two stage high rate process to provide complete sewage treatment. Four mineral and four plastic primary filter media and two plastic secondary filter media were studied. The hydraulic loading applied to the primary plastic media (11.2 m3 /m3 .d) was twice that applied to the mineral media. The plastic media removed an average around 66 percent and the mineral media around 73 percent of the BOD applied when the 90 percentile BOD concentration was 563 mg/1. At a hydraulic loading of 4 m3 /m3 .d the secondary filters removed most of the POD from partially settled primary filter effluents, with one secondary effluent satisfying a 25 mg/1 BOD and 30 mg/1 SS standard. No significant degree of nitrification was achieved. Fungi dominated the biological film of the primary filters, with invertebrate grazers having little influence on film levels. Ponding did not arise, and modular media supported lower film levels than random-fill types. Secondary filter film levels were low, being dominated by bacteria. The biological loading applied to the filters was related to sludge dewaterability, with the most readily conditionable sludges produced by filters supporting heavy film. Sludges produced by random-fill media could be dewatered as readily as those produced by low rate filters treating the same sewage. Laboratory scale studies showed a relationship between log effluent BOD and nitrification achieved by biological filters. This relationship and the relationship between BOD load applied and removed observed in all filter media could he used to optimise operating conditions required in biological filters to achieve given effluent BOD and ammoniacal nitrogen standards.

Marketing in indigenous and Asian small firms in the West Midlands.

This study is concerned with examining the application of marketing during the start-up, development and growth of small firms in the West Midlands. As an exploratory study, it provides evidence to support the central hypothesis of the thesis that whilst many small firms have the potential to progress through to the successful growth stage of development, they fail to do so because of their owner-managers' orientation towards production and selling and because they do not apply formal marketing during the initial stages of business development. A comparative approach to studying marketing in indigenous and Asian firms is adopted in an attempt to fill a gap in the literature on the characteristics and differences in the formation and development processes of these two groups of enterprises. The study has three main objectives and is based on qualitative research techniques of in-depth interviews, case studies and longitudinal studies among sixty-six firms representing the key activities of the small firms sector of the local economy. Firstly, it investigates owner-managers' orientation in developing and managing new and established businesses and explores the sources of, and changes in their orientation during the various stages of development. Secondly, it assesses the owner-manager's awareness and understanding of what constitutes the marketing function and investigates what aspects of marketing are applied during the different stages of business development. Finally, the study monitors and evaluates the outcomes and implications of applying formal marketing techniques in a small sample of firms over a period of two years. The thesis concludes by using the findings of the study to contribute additions to existing models of growth and by proposing new models of evolution and application of marketing in small firms.

Third sector organisations and social welfare : a study of the role of social service and charitable organisations in Taiwan

This study is about the role and operation of ‘third sector’ organisations (TSOs) within the Taiwanese social welfare context. TSOs have increased dramatically and become actively involved in social service provision. This phenomenon has not only had significant impact on the development and operation of TSOs in Taiwan but it is also of increasing interest to public policy academics. The latter are especially interested in the implications for the government-third sector relationship. This research examines the reasons why TSOs have been established, why they actively participate in social service provision, and their role and operation within the social welfare context of Taiwan. The study has both quantitative and qualitative data. It sampled ‘social service’ and ‘charitable’ organisations (SSCOs), which are the main type of TSOs in Taiwan, to examine their role, operation and interaction with government. Questionnaires were mailed to collect quantitative data first. After the quantitative data were collected and analysed, semi-structured interviews were undertaken to collect qualitative data. The study found that TSOs in Taiwan exist in a highly institutionalised environment, which is affected by traditional Confucian ideas and contemporary Western ideas such as social justice and civil rights. The rapid growth of TSOs has a strong connection with the desire to fill social service gaps left by government and family. TSOs mainly play the role of service provider rather than that of advocate. They cooperate with government in social service provision and have developed different types of symbiotic relationships with government. A ‘resonance effect’ between government and TSOs was also found as they implement social policy.

Strategic positioning in voluntary and non-profit organizations in the UK : exploring the experiences of British charitable organizations:Exploring the experiences of British Charitable Organizations

This thesis explores the strategic positioning [SP] activities of charitable organizations [COs] within the wider sector of voluntary and non-profit organizations [VNPOs] in the UK. Despite the growing interest in SP for British COs in an increasingly competitive operating environment and changing policy context, there is lack of research in mainstream marketing/strategic management studies on this topic for charities, whilst the specialist literature on VNPOs has neglected the study of SP. The thesis begins with an extended literature review of the concept of positioning in both commercial [for-profit] and charitable organizations. It concludes that the majority of theoretical underpinnings of SP that are prescribed for COs have been derived from the commercial strategy/marketing literature. There is currently a lack of theoretical and conceptual models that can accommodate the particular context of COs and guide strategic positioning practice in them. The research contained in this thesis is intended to fill some of these research gaps. It combines an exploratory postal survey and four cross-sectional case studies to describe the SP activities of a sample of general welfare and social care charities and identifies the key factors that influence their choice of positioning strategies [PSs]. It concludes that charitable organizations have begun to undertake SP to differentiate their organizations from other charities that provide similar services. Their PSs have both generic features, and other characteristics that are unique to them. A combination of external environmental and organizational factors influences their choice of PSs. A theoretical model, which depicts these factors, is developed in this research. It highlights the role of governmental influence, other external environmental forces, the charity’s mission, organizational resources, and influential stakeholders in shaping the charity’s PS. This study concludes by considering the theoretical and managerial implications of the findings on the study of charitable and non-profit organizations.

Phonophoresis and topical drug delivery

In this study, investigations into phonophoresis were conducted by employing 3 distinct in vitro models. The aim of the first model was to evaluate the effect of ultrasound on the migration rate of different classes of molecules through agar gel. The derived data suggested that small, relatively hydrophobic molecules are more susceptible to ultrasound-enhanced diffusion through the water-filled channels of the agar gel. The application of heat alone increased drug migration by a similar magnitude as the ultrasound, indicating that ultrasonic heating directly increases the thermodynamic potential for diffusion. In the second experimental system, whole rat skin was pre-sonicated and then examined for changes in its barrier properties. At high intensities (1 to 2W cm-2), ultrasonic waves irreversibly compromised the barrier properties of the skin, following the general patterns described in the literature reports. At low intensities (< 1W cm-2), ultrasound discharged sebum from the sebaceous glands so as to fill much of the hair follicle shafts. This entirely novel phenomenon is probably produced by the mechanical effects of the beam. The deposition of sebaceous lipids within the hair follicle shafts can mean that this absorption pathway is blocked for hydrophilic molecules that penetrate via this route. Consequently, this phenomenon can be utilised as a probe to measure the relative follicular contribution to total penetration for these molecules. In the final phonophoresis model, modified Franz cells were employed in order to assess the ultrasound effect on the concurrent transdermal permeation of various molecules through whole rat skin. For the most lipophilic agent tested, the rate-limiting step of absorption was partitioning from the stratum corneum into the viable epidermis. Sonication did not accelerate this step.

An appraisal of employment problems and policies at the micro-economic level

This study has concentrated on the development of an impact simulation model for use at the sub-national level. The necessity for the development of this model was demonstrated by the growth of local economic initiatives during the 1970's, and the lack of monitoring and evaluation exercise to assess their success and cost-effectiveness. The first stage of research involved the confirmation that the potential for micro-economic and spatial initiatives existed. This was done by identifying the existence of involuntary structural unemployment. The second stage examined the range of employment policy options from the macroeconomic, micro-economic and spatial perspectives, and focused on the need for evaluation of those policies. The need for spatial impact evaluation exercise in respect of other exogenous shocks, and structural changes was also recognised. The final stage involved the investigation of current techniques of evaluation and their adaptation for the purpose in hand. This led to a recognition of a gap in the armoury of techniques. The employment-dependency model has been developed to fill that gap, providing a low-budget model, capable of implementation at the small area level and generating a vast array of industrially disaggregate data, in terms of employment, employment-income, profits, value-added and gross income, related to levels of United Kingdom final demand. Thus providing scope for a variety of impact simulation exercises.

Settlement of fill with particular reference to opencast coal mining sites

A study of information available on the settlement characteristics of backfill in restored opencast coal mining sites and other similar earthworks projects has been undertaken. In addition, the methods of opencast mining, compaction controls, monitoring and test methods have been reviewed. To consider and develop the methods of predicting the settlement of fill, three sites in the West Midlands have been examined; at each, the backfill had been placed in a controlled manner. In addition, use has been made of a finite element computer program to compare a simple two-dimensional linear elastic analysis with field observations of surface settlements in the vicinity of buried highwalls. On controlled backfill sites, settlement predictions have been accurately made, based on a linear relationship between settlement (expressed as a percentage of fill height) against logarithm of time. This `creep' settlement was found to be effectively complete within 18 months of restoration. A decrease of this percentage settlement was observed with increasing fill thickness; this is believed to be related to the speed with which the backfill is placed. A rising water table within the backfill is indicated to cause additional gradual settlement. A prediction method, based on settlement monitoring, has been developed and used to determine the pattern of settlement across highwalls and buried highwalls. The zone of appreciable differential settlement was found to be mainly limited to the highwall area, the magnitude was dictated by the highwall inclination. With a backfill cover of about 15 metres over a buried highwall the magnitude of differential settlement was negligible. Use has been made of the proposed settlement prediction method and monitoring to control the re-development of restored opencase sites. The specifications, tests and monitoring techniques developed in recent years have been used to aid this. Such techniques have been valuable in restoring land previously derelict due to past underground mining.

Metaphor in (Arabic-into-English) translation with specific reference to metaphorical concepts and expressions in political discourse

Cognitive linguistics scholars argue that metaphor is fundamentally a conceptual process of mapping one domain of experience onto another domain. The study of metaphor in the context of Translation Studies has not, unfortunately, kept pace with the discoveries about the nature and role of metaphor in the cognitive sciences. This study aims primarily to fill part of this gap of knowledge. Specifically, the thesis is an attempt to explore some implications of the conceptual theory of metaphor for translation. Because the study of metaphor in translation is also based on views about the nature of translation, the thesis first presents a general overview of the discipline of Translation Studies, describing the major models of translation. The study (in Chapter Two) then discusses the major traditional theories of metaphor (comparison, substitution and interaction theories) and shows how the ideas of those theories were adopted in specific translation studies of metaphor. After that, the study presents a detailed account of the conceptual theory of metaphor and some hypothetical implications for the study of metaphor in translation from the perspective of cognitive linguistics. The data and methodology are presented in Chapter Four. A novel classification of conceptual metaphor is presented which distinguishes between different source domains of conceptual metaphors: physical, human-life and intertextual. It is suggested that each source domain places different demands on translators. The major sources of the data for this study are (1) the translations done by the Foreign Broadcasting Information Service (FBIS), which is a translation service of the Central Intelligence Agency (CIA) in the United Sates of America, of a number of speeches by the Iraqi president Saddam Hussein during the Gulf Crisis (1990-1991) and (2) official (governmental) Omani translations of National Day speeches of Sultan Qaboos bin Said of Oman.

Discrete element modelling of a rotating drum and drum granulation

This thesis reports the results of DEM (Discrete Element Method) simulations of rotating drums operated in a number of different flow regimes. DEM simulations of drum granulation have also been conducted. The aim was to demonstrate that a realistic simulation is possible, and further understanding of the particle motion and granulation processes in a rotating drum. The simulation model has shown good qualitative and quantitative agreement with other published experimental results. A two-dimensional bed of 5000 disc particles, with properties similar to glass has been simulated in the rolling mode (Froude number 0.0076) with a fractional drum fill of approximately 30%. Particle velocity fields in the cascading layer, bed cross-section, and at the drum wall have shown good agreement with experimental PEPT data. Particle avalanches in the cascading layer have been shown to be consistent with single layers of particles cascading down the free surface towards the drum wall. Particle slip at the drum wall has been shown to depend on angular position, and ranged from 20% at the toe and shoulder, to less than 1% at the mid-point. Three-dimensional DEM simulations of a moderately cascading bed of 50,000 spherical elastic particles (Froude number 0.83) with a fractional fill of approximately 30% have also been performed. The drum axis was inclined by 50 to the horizontal with periodic boundaries at the ends of the drum. The mean period of bed circulation was found to be 0.28s. A liquid binder was added to the system using a spray model based on the concept of a wet surface energy. Granule formation and breakage processes have been demonstrated in the system.