Comparative proteomic analysis of GS-NSO murine myeloma cell lines with varying recombinant monoclonal antibody production rate

We have employed an inverse engineering strategy based on quantitative proteome analysis to identify changes in intracellular protein abundance that correlate with increased specific recombinant monoclonal antibody production (qMab) by engineered murine myeloma (NSO) cells. Four homogeneous NSO cell lines differing in qMab were isolated from a pool of primary transfectants. The proteome of each stably transfected cell line was analyzed at mid-exponential growth phase by two-dimensional gel electrophoresis (2D-PAGE) and individual protein spot volume data derived from digitized gel images were compared statistically. To identify changes in protein abundance associated with qMab clatasets were screened for proteins that exhibited either a linear correlation with cell line qMab or a conserved change in abundance specific only to the cell line with highest qMab. Several proteins with altered abundance were identified by mass spectrometry. Proteins exhibiting a significant increase in abundance with increasing qMab included molecular chaperones known to interact directly with nascent immunoglobulins during their folding and assembly (e.g., BiP, endoplasmin, protein disulfide isomerase). 2D-PAGE analysis showed that in all cell lines Mab light chain was more abundant than heavy chain, indicating that this is a likely prerequisite for efficient Mab production. In summary, these data reveal both the adaptive responses and molecular mechanisms enabling mammalian cells in culture to achieve high-level recombinant monoclonal antibody production. (C) 2004 Wiley Periodicals, Inc.

On the optimal ratio of heavy to light chain genes for efficient recombinant antibody production by CHO cells

Monoclonal antibodies (Mab) are heterotetramers consisting of an equimolar ratio of heavy chain (HC) and light chain (LC) polypeptides. Accordingly, most recombinant Mab expression systems utilize an equimolar ratio of heavy chain (he) to light chain (lc) genes encoded on either one or two plasmids. However, there is no evidence to suggest that this gene ratio is optimal for stable or transient production of recombinant Mab. In this study we have determined the optimal ratio of hc:lc genes for production of a recombinant IgG(4) Mab, cB72.3, by Chinese hamster ovary (CHO) cells using both empirical and mathematical modeling approaches. Polyethyleneimine-mediated transient expression of cB72.3 at varying ratios of hc:lc genes encoded on separate plasmids yielded an optimal Mab titer at a hc:lc gene ratio of 3:2; a conclusion confirmed by separate mathematical modeling of the Mab folding and assembly process using transient expression data. On the basis of this information, we hypothesized that utilization of he genes at low hc:lc gene ratios is more efficient. To confirm this, cB72.3 Mab was transiently produced by CHO cells at constant he and varying lc gene dose. Under these conditions, Mab yield was increased with a concomitant increase in lc gene dose. To determine if the above findings also apply to stably transfected CHO cells producing recombinant Mab, we compared the intra- and extracellular ratios of HC and LC polypeptides for three GS-CHO cells lines transfected with a 1:1 ratio of hc:lc genes and selected for stable expression of the same recombinant Mab, cB72.3. Intra- and extracellular HC:LC polypeptide ratios ranged from 1:2 to 1:5, less than that observed on transient expression of the same Mab in parental CHO cells using the same vector. In conclusion, our data suggest that the optimal ratio of hc:lc genes used for transient and stable expression of Mab differ. In the case of the latter, we infer that optimal Mab production by stably transfected cells represents a compromise between HC abundance limiting productivity and the requirement for excess LC to render Mab folding and assembly more efficient.

Summarizing level-two topological relations in large spatial datasets

Summarizing topological relations is fundamental to many spatial applications including spatial query optimization. In this article, we present several novel techniques to effectively construct cell density based spatial histograms for range (window) summarizations restricted to the four most important level-two topological relations: contains, contained, overlap, and disjoint. We first present a novel framework to construct a multiscale Euler histogram in 2D space with the guarantee of the exact summarization results for aligned windows in constant time. To minimize the storage space in such a multiscale Euler histogram, an approximate algorithm with the approximate ratio 19/12 is presented, while the problem is shown NP-hard generally. To conform to a limited storage space where a multiscale histogram may be allowed to have only k Euler histograms, an effective algorithm is presented to construct multiscale histograms to achieve high accuracy in approximately summarizing aligned windows. Then, we present a new approximate algorithm to query an Euler histogram that cannot guarantee the exact answers; it runs in constant time. We also investigate the problem of nonaligned windows and the problem of effectively partitioning the data space to support nonaligned window queries. Finally, we extend our techniques to 3D space. Our extensive experiments against both synthetic and real world datasets demonstrate that the approximate multiscale histogram techniques may improve the accuracy of the existing techniques by several orders of magnitude while retaining the cost efficiency, and the exact multiscale histogram technique requires only a storage space linearly proportional to the number of cells for many popular real datasets.

The effects of nitrogen application and assimilate availability on engorged pollen production and spikelet sterility in rice

Increased rates of nitrogen fertilizer application lead to increased spikelet sterility. A field experiment was conducted to investigate the effects on engorged pollen production and spikelet sterility, of nitrogen and assimilate availability during microspore development, in two rice cultivars (Doongara and Amaroo) grown under two different water depths. Despite the temperature not being low enough during microspore development to cause spikelet sterility, the number of engorged pollen grains was lower in cv. Doongara than in cv. Amaroo. Nitrogen application decreased the number of engorged pollen grains per anther through increased spikelet density. Nitrogen application increased spikelet sterility as a result of increased panicle density showing pronounced indirect effect of N on spikelet sterility. Engorged pollen number was also closely related (r = -0.636*) to the nitrogen content of the leaf blade, indicating a direct negative effect of plant N status on engorged pollen production. The results suggest that the intrinsic pollen producing ability is the key element in the difference in cold tolerance between the two cultivars, particularly under high N rates. Opening the canopy for increased solar radiation interception by the treated plants increased the level of engorged pollen, indicating the importance of immediate assimilate availability for engorged pollen production. Shading reduced crop growth rate, but did not effect engorged pollen production. There was no effect of variation in assimilates production on spikelet sterility.

COMUNICAÇÃO PÚBLICA: UMA LEITURA CRÍTICA ENTRE O INSTITUCIONAL E O MERCADOLÓGICO

A pesquisa ora apresentada discute os processos comunicacionais e de gestão no desenvolvimento de atividades de Comunicação Pública. Sobretudo aqueles que sejam capazes de alcançar um elevado nível de qualidade em prol da eficácia na prestação de serviços ao cidadão-consumidor. Transitando entre a comunicação institucional e a mercadológica, em referência à linha de pesquisa de mesmo nome constante do Programa de Pós-graduação de Comunicação Social da Universidade, o trabalho em questão aborda tópicos como o estudo de boas práticas e aspectos interessantes sobre processos, políticas, estratégias e ações na gestão de competências da comunicação nas organizações. Para isso, em termos de metodologia, são utilizados elementos bibliográficos e documentais, além de entrevistas abertas com os gestores do setor que conformam exemplos de comunicação organizacional em duas instituições. Esses procedimentos verificados durante a pesquisa no processo comunicacional das organizações estudadas se correlacionam a uma contextualização histórica de conceitos, teorias e métodos voltados à realidade da sociedade contemporânea. Análise justificada em que pese uma abordagem a respeito de princípios que possam ser visualizados na práxis comunicacional, como sustentabilidade, transparência, governança corporativa e gestão da comunicação em uma ambiência tecnossocial de produção simbólica de conteúdos (informações, produtos, serviços, marcas, imagem etc.). A análise dessa dinâmica como um todo objetiva abordar, ainda, as implicações sociais do processo de planejamento em Comunicação e do ato comunicativo em si, que nos faz perceber de que forma as organizações públicas observam a relação (discurso versus prática) com seus clientes no acesso a informações, produtos ou serviços.

Retail restructuring and consumer choice 2:understanding consumer choice at the household level

This paper complements the preceding one by Clarke et al, which looked at the long-term impact of retail restructuring on consumer choice at the local level. Whereas the previous paper was based on quantitative evidence from survey research, this paper draws on the qualitative phases of the same three-year study, and in it we aim to understand how the changing forms of retail provision are experienced at the neighbourhood and household level. The empirical material is drawn from focus groups, accompanied shopping trips, diaries, interviews, and kitchen visits with eight households in two contrasting neighbourhoods in the Portsmouth area. The data demonstrate that consumer choice involves judgments of taste, quality, and value as well as more ‘objective’ questions of convenience, price, and accessibility. These judgments are related to households’ differential levels of cultural capital and involve ethical and moral considerations as well as more mundane considerations of practical utility. Our evidence suggests that many of the terms that are conventionally advanced as explanations of consumer choice (such as ‘convenience’, ‘value’, and ‘habit’) have very different meanings according to different household circumstances. To understand these meanings requires us to relate consumers’ at-store behaviour to the domestic context in which their consumption choices are embedded. Bringing theories of practice to bear on the nature of consumer choice, our research demonstrates that consumer choice between stores can be understood in terms of accessibility and convenience, whereas choice within stores involves notions of value, price, and quality. We also demonstrate that choice between and within stores is strongly mediated by consumers’ household contexts, reflecting the extent to which shopping practices are embedded within consumers’ domestic routines and complex everyday lives. The paper concludes with a summary of the overall findings of the project, and with a discussion of the practical and theoretical implications of the study.

High level data fusion

We address the question of how to obtain effective fusion of identification information such that it is robust to the quality of this information. As well as technical issues data fusion is encumbered with a collection of (potentially confusing) practical considerations. These considerations are described during the early chapters in which a framework for data fusion is developed. Following this process of diversification it becomes clear that the original question is not well posed and requires more precise specification. We use the framework to focus on some of the technical issues relevant to the question being addressed. We show that fusion of hard decisions through use of an adaptive version of the maximum a posteriori decision rule yields acceptable performance. Better performance is possible using probability level fusion as long as the probabilities are accurate. Of particular interest is the prevalence of overconfidence and the effect it has on fused performance. The production of accurate probabilities from poor quality data forms the latter part of the thesis. Two approaches are taken. Firstly the probabilities may be moderated at source (either analytically or numerically). Secondly, the probabilities may be transformed at the fusion centre. In each case an improvement in fused performance is demonstrated. We therefore conclude that in order to obtain robust fusion care should be taken to model the probabilities accurately; either at the source or centrally.

Planning and decision-making for energy recovery from waste at the sub-regional level

This research project has developed a novel decision support system using Geographical Information Systems and Multi Criteria Decision Analysis and used it to develop and evaluate energy-from-waste policy options. The system was validated by applying it to the UK administrative areas of Cornwall and Warwickshire. Different strategies have been defined by the size and number of the facilities, as well as the technology chosen. Using sensitivity on the results from the decision support system, it was found that key decision criteria included those affected by cost, energy efficiency, transport impacts and air/dioxin emissions. The conclusions of this work are that distributed small-scale energy-from-waste facilities score most highly overall and that scale is more important than technology design in determining overall policy impact. This project makes its primary contribution to energy-from-waste planning by its development of a Decision Support System that can be used to assist waste disposal authorities to identify preferred energy-from-waste options that have been tailored specifically to the socio-geographic characteristics of their jurisdictional areas. The project also highlights the potential of energy-from-waste policies that are seldom given enough attention to in the UK, namely those of a smaller-scale and distributed nature that often have technology designed specifically to cater for this market.

Strategic planning as communicative process

This thesis looks at the construction of a strategic plan within a British university (Unico). After a change in leadership, the well-known strategic planning sequence was adopted to set directions according to Unico’s three Missions, followed by the development of respective goals and measures. The evolving strategic content coincided with the development of Unico’s strategic plan. I was able to follow Unico’s planning efforts over 10 months, from first planning meeting to completion of its strategic plan. The main data source provided non-participant observation (n = 25) and ten versions of Unico’s strategic plan. Additionally, seventy-six interviews were held with participants at various points. In order to examine the strategic plan’s construction, I reconceptualised strategic planning as a communicative process consisting of oral talk and written text. Through this interplay strategic planning activities come in to being. Such reconceptualisation provided a conceptual framework to study the in situ interactions without neglecting contextual characteristics embedding the communicative process. Strategic plans are currently seen as promoting inflexibility and reinforcing the institutional nature of formal strategic planning. Adopting dialogism, as advocated by Bakhtin and Ricoeur, this research provides novel insights into the dialogic of strategy talk and strategy text, such as a strategic plan. Findings illustrated that a strategic plan production cycle provided a meaning making platform for its participants. Through recurrently amending the plan, its content became increasingly specific while at the same time reflecting agreed terminology. This thesis offers an alternative view on strategic planning, elaborates on the strategy-as-practice perspective, focusing on the under-explored area of individuals’ interactions at the micro level, and elaborates on the dialogic of text and agency/conversation, distinguishing between talk and text.

Can production subsidies explain China's export performance? Evidence from firm-level data

This paper analyses the relationship between production subsidies and firms’ export performance using a very comprehensive and recent firm-level database and controlling for the endogeneity of subsidies. It documents robust evidence that production subsidies stimulate export activity at the intensive margin, although this effect is conditional on firm characteristics. In particular, the positive relationship between subsidies and the intensive margin of exports is strongest among profit-making firms, firms in capital-intensive industries, and those located in non-coastal regions. Compared to firm characteristics, the extent of heterogeneity across ownership structure (SOEs, collectives, and privately owned firms) proves to be relatively less important.