A chicken bioreactor for efficient production of functional cytokines

The global market for protein drugs has the highest compound annual growth rate of any pharmaceutical class but their availability, especially outside of the US market, is compromised by the high cost of manufacture and validation compared to traditional chemical drugs. Improvements in transgenic technologies allow valuable proteins to be produced by genetically-modified animals; several therapeutic proteins from such animal bioreactors are already on the market after successful clinical trials and regulatory approval. Chickens have lagged behind mammals in bioreactor development, despite a number of potential advantages, due to the historic difficulty in producing transgenic birds, but the production of therapeutic proteins in egg white of transgenic chickens would substantially lower costs across the entire production cycle compared to traditional cell culture-based production systems. This could lead to more affordable treatments and wider markets, including in developing countries and for animal health applications. Here we report the efficient generation of new transgenic chicken lines to optimize protein production in eggs. As proof-of-concept, we describe the expression, purification and functional characterization of three pharmaceutical proteins, the human cytokine interferon α2a and two species-specific Fc fusions of the cytokine CSF1. Our work optimizes and validates a transgenic chicken system for the cost-effective production of pure, high quality, biologically active protein for therapeutics and other applications

Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis

For the improved production of vaccines and therapeutic proteins, a detailed understanding of the metabolic dynamics during batch or fed-batch production is requested. To study the new human cell line AGE1.HN, a flexible metabolic flux analysis method was developed that is considering dynamic changes in growth and metabolism during cultivation. This method comprises analysis of formation of cellular components as well as conversion of major substrates and products, spline fitting of dynamic data and flux estimation using metabolite balancing. During batch cultivation of AGE1.HN three distinct phases were observed, an initial one with consumption of pyruvate and high glycolytic activity, a second characterized by a highly efficient metabolism with very little energy spilling waste production and a third with glutamine limitation and decreasing viability. Main events triggering changes in cellular metabolism were depletion of pyruvate and glutamine. Potential targets for the improvement identified from the analysis are (i) reduction of overflow metabolism in the beginning of cultivation, e.g. accomplished by reduction of pyruvate content in the medium and (ii) prolongation of phase 2 with its highly efficient energy metabolism applying e.g. specific feeding strategies. The method presented allows fast and reliable metabolic flux analysis during the development of producer cells and production processes from microtiter plate to large scale reactors with moderate analytical and computational effort. It seems well suited to guide media optimization and genetic engineering of producing cell lines

Treatment Patterns and Use of Resources in Patients With Tuberous Sclerosis Complex: Insights From the TOSCA Registry

Tuberous Sclerosis Complex (TSC) is a rare autosomal-dominant disorder caused by mutations in the TSC1 or TSC2 genes. Patients with TSC may suffer from a wide range of clinical manifestations; however, the burden of TSC and its impact on healthcare resources needed for its management remain unknown. Besides, the use of resources might vary across countries depending on the country-specific clinical practice. The aim of this paper is to describe the use of TSC-related resources and treatment patterns within the TOSCA registry. A total of 2,214 patients with TSC from 31 countries were enrolled and had a follow-up of up to 5 years. A search was conducted to identify the variables containing both medical and non-medical resource use information within TOSCA. This search was performed both at the level of the core project as well as at the level of the research projects on epilepsy, subependymal giant cell astrocytoma (SEGA), lymphangioleiomyomatosis (LAM), and renal angiomyolipoma (rAML) taking into account the timepoints of the study, age groups, and countries. Data from the quality of life (QoL) research project were analyzed by type of visit and age at enrollment. Treatments varied greatly depending on the clinical manifestation, timepoint in the study, and age groups. GAB Aergics were the most prescribed drugs for epilepsy, and mTOR inhibitors are dramatically replacing surgery in patients with SEGA, despite current recommendations proposing both treatment options. mTOR inhibitors are also becoming common treatments in rAML and LAM patients. Forty-two out of the 143 patients (29.4%) who participated in the QoL research project reported inpatient stays over the last year. Data from non-medical resource use showed the critical impact of TSC on job status and capacity. Disability allowances were more common in children than adults (51.1% vs 38.2%). Psychological counseling, social services and social worker services were needed by <15% of the patients, regardless of age. The long-term nature, together with the variability in its clinical manifestations, makes TSC a complex and resource-demanding disease. The present study shows a comprehensive picture of the resource use implications of TSC

Providing Rate Guarantees For Internet Application Traffic Across ATM Networks

The Transmission Control Protocol/Internet Protocol (TCP/IP) protocol suite is the standard requirement for all applications that need to communicate over the Internet. As TCP/IP applications are unable to specify the QoS parameters needed for most of the Asynchronous Transfer Mode (ATM) services, they tend to use the Unspecified Bit Rate (UBR) service category when running across ATM networks. The UBR service utilises any bandwidth which is left unused by the rest of the ATM services. This has led the ATM Forum&apos;s Traffic Management group to define a new service category called Guaranteed Frame Rate (GFR). GFR is intended to provide minimum cell rate guarantees and fair access to excess bandwidth, left over from higher-priority services. This article gives first a tutorial overview of GFR and then presents a survey of the research work which has been carried out towards the design and implementation of associated ATM switch mechanisms

Between the legacy of nation-state and forces of globalisation : Turkey’s management of mixed migration flows

Turkey, at the crossroads of Europe, Middle East and Asia, has confronted with mounting pressures of mixed migration flows in recent decades. This paper aims to explore Turkey’s contemporary approach to migration management by focusing on the adoption of the country’s first comprehensive immigration law (Law on Foreigners and International Protection) and the signing of the readmission agreement with the European Union in 2013. This incorporates an analysis of both policy continuities and changes in migration management in Turkey, while also providing an understanding of the interplay between internal and external factors, namely internationalisation and Europeanisation processes and the responsiveness of domestic actors to such pressures. The paper argues that migration policies driven solely by state-centric concerns are becoming increasingly inefficient in responding to the challenges caused by interlinked pressures of globalisation and multi-layered migratory flows. As Turkey’s role as a transit and receiving country grows, issues of international migration, and irregular migration in particular, are becoming dynamic topics in defining its role in a globalised world and as well as the trajectory of its relations with the EU

Exploring the determinants and dimensionality of SME eCollaboration trust

SMEs are now an important and integral part of a country\u27s economy and have been acknowledged as being very different to their larger counterparts (Street and Meister, 2004). The economic viability of many economies is dependent on the survival and future growth of this sector. However, small firms are increasingly exposed to competition from larger, resource-rich firms; therefore, it is not uncommon for SMEs to become involved in collaborations to grow their business, increase their competitive position and leverage opportunities presented by new technologies (e-Collaboration) (Lawson and Hoi, 2007). Although emerging research suggests that e-Collaboration is vital for the future survival of small to medium-sized businesses (Henttonen and Blomqvist, 2005; Lawson and Hoi, 2007), it also stresses that dealing with issues surrounding trust becomes of paramount importance. This chapter presents a discussion on the role and importance of trust in collaborative SME environments and provides a starting point from which to develop an understanding of SME e-Collaboration trust, its determinants and dimensionality