Optimisation of retroviral production systems for gene therapy applications

Retroviral vectors are a promising tool for gene therapy. However, there are two major problems to overcome if a viable commercial production process is to be established. These are the instability of virus particles and the low virus titres. The characteristics of the producer cells were determined in batch culture, semicontinuous culture and semi-continuous culture at 32°C. Additionally, cell attachment, growth and virus production on various macroporous microcarriers was assessed under static and stirred conditions. Alternative strategies for the cultivation of cells were also investigated. These included spinner basket, packed bed and spinner flask cultures with semi-continuous feeding and packed bed, fixed bed, fluidised bed and stirred tank cultures with continuous perfusion of culture medium. Of these the fixed bed bioreactor had the highest cell specific productivity and was capable of running for 28 days. The fluidised bed bioreactor had the highest reactor productivity, due to the higher cell number. Optimisation of culture medium was performed with regard to serum concentration. The greatest production was observed at an initial serum concentration of 2.5% (v/v). The findings in this thesis will assist the development of an efficient method for the production of clinical grade retro viral vectors for gene therapy applications

Review of offshore cable reliability metrics

Reliable cable systems are essential for offshore wind operation. Industry trends have led to a large number of offshore cable connections being installed recently, with 11027 MW of offshore wind connected at the end of 2015 compared to just 2955 MW in 2010 [1]. Despite the increase in connections, the publically available reliability data in this area is almost non-existent. With several connections in planning of both similar and increasing lengths it is essential to better understand these metrics. A review of published reliability data was undertaken in order to populate a database which is presented in this paper. This data focusses on a number of connection types including both AC and DC connections across a number of cable ratings and configurations. From this database it is confirmed that reliability figures currently being used across the literature generally conform to those currently being experienced in the offshore wind industry. However it is established that failure rates taken from some reports are not accurate as the technology and environments these are calculated from are typically different from those used in offshore wind farm connections. This information is collated and converted into reliability metrics in order for comparisons to be made. Analysis of the cost of an outage experienced by a windfarm is also carried out in this paper. The results of which establish that the revenue lost from a cable failure could potentially be substantial. The findings in this paper would also suggest a greater risk of failure in the early life of a windfarm and as such a greater potential cost associated with this risk. It is important to have a better understanding of offshore renewable energy cable connections as the reliability of a cable has a significant impact on the Levelised Cost of Energy. With a greater understanding of the metrics investors can make more informed decisions with respect to the technology that is installed as well as the importance of the installation process itself, due diligence on subsequent OFTO asset purchases and the maintenance plans that have been outlined for the connection

A Study Of HRMs Response To The Events Of 9/11

The 9/11 terrorist attacks leveled the twin towers of New Yorks World Trade Center, caused serious damage to the Pentagon, and disrupted the national economy. The response by corporate organiza-tions, hardest hit by the events of 9/11, has turned the familiar phrase business as usual into an anachronism. Effective human resource management (HRM) practices are critical for any organization to appropriately respond and recover from catastrophic events, such as those of 9/11. In the aftermath of 9/11, corporate organizations have recognized the need to change many of their HRM practices. Possible explanations for recent changes in HRM practices are twofold: 1) as precautionary measures intended to avert man-made disasters, and 2) to enhance the organizations capability to immediately and effectively respond should a disaster strike. Irrespective of the reasons organizations are changing their HRM practices, they are placing more importance than ever on the need to optimally utilize and safeguard their most valued asset employees. Exploratory research on the initial responses by corporate organizations to the events of 9/11 reveals: 1) that many did not have disaster plans in place, and 2) that an even larger number were poorly prepared to deal with the disasters immediate impact on employees, particularly the negative psychological consequences engendered by such a disaster. This exploratory paper culminates in a proposed HRM Practices Disaster Recovery Diagnostic Tool. This tool would allow an organization to assess the adequacy of their HRM practices in responding to man-made disasters

Construction of Large Period Symplectic Maps by Interpolative Methods

The goal is to construct a symplectic evolution map for a large section of an accelerator, say a full turn of a large ring or a long wiggler. We start with an accurate tracking algorithm for single particles, which is allowed to be slightly non-symplectic. By tracking many particles for a distance S one acquires sufficient data to construct the mixed-variable generator of a symplectic map for evolution over S, given in terms of interpolatory functions. Two ways to find the generator are considered: (1) Find its gradient from tracking data, then the generator itself as a line integral. (2) Compute the action integral on many orbits. A test of method (1) has been made in a difficult example: a full turn map for an electron ring with strong nonlinearity near the dynamic aperture. The method succeeds at fairly large amplitudes, but there are technical difficulties near the dynamic aperture due to oddly shaped interpolation domains. For a generally applicable algorithm we propose method (2), realized with meshless interpolation methods

Calibration uncertainty in molecular dating analyses: there is no substitute for the prior evaluation of time priors

Calibration is the rate-determining step in every molecular clock analysis and, hence, considerable effort has been expended in the development of approaches to distinguish good from bad calibrations. These can be categorized into a priori evaluation of the intrinsic fossil evidence, and a posteriori evaluation of congruence through cross-validation. We contrasted these competing approaches and explored the impact of different interpretations of the fossil evidence upon Bayesian divergence time estimation. The results demonstrate that a posteriori approaches can lead to the selection of erroneous calibrations. Bayesian posterior estimates are also shown to be extremely sensitive to the probabilistic interpretation of temporal constraints. Furthermore, the effective time priors implemented within an analysis differ for individual calibrations when employed alone and in differing combination with others. This compromises the implicit assumption of all calibration consistency methods, that the impact of an individual calibration is the same when used alone or in unison with others. Thus, the most effective means of establishing the quality of fossil-based calibrations is through a priori evaluation of the intrinsic palaeontological, stratigraphic, geochronological and phylogenetic data. However, effort expended in establishing calibrations will not be rewarded unless they are implemented faithfully in divergence time analyses

Prospectus, October 23, 1970

TEACHING COMPUTER USES TUTOR, INQUIRY; New College To Be Extension Of Old; Scheduling System Is Changed; Problems Of Our Times; Letters To The Editor; Bull Page: All Organizations, Wit N\u27 Wisdom, Athletics Committee, Yearbook, Powder Puff Football, The Rainmaker Is Cast, Chi Gamma Iota, Cinema, Student Government, Phi Beta Lambda, IOC, Vets Meeting; I\u27m For Real: October\u27s Intercollegiate Athlete; Golfers- In Full Swing; Coaches Corner; About The Artisthttps://spark.parkland.edu/prospectus_1970/1023/thumbnail.jp

Failure rates of offshore wind transmission systems

In the offshore wind industry, failures are often costlier than those experienced onshore. Through examination of the literature, it is clear that failures occurring in offshore transmission systems are not well documented. As a result of this, many developers and other parties involved in the planning processes associated with offshore wind farms will defer back to existing reliability metrics in the public domain. This article presents a review of European offshore wind farm transmission failures based on fusing information from multiple public domain sources. The results highlight both the spread of the reliability performance of these assets and the reliability performance over time. The results also reinforce the industry view that installation practices could lead to low reliability in the initial years of operation, resulting in increased repair costs and decreased revenue for wind farm owners and operators. The information collated in the review is also compared to metrics from across the literature to evaluate the difference in forecasted failure rates to those experienced within the industry. In general, it is found that the experienced failure rates are subject to a much higher spread in practice than those published until now

Culture of equine fibroblast-like synoviocytes on synthetic tissue scaffolds towards meniscal tissue engineering: a preliminary cell-seeding study

Introduction. Tissue engineering is a new methodology for addressing meniscal injury or loss. Synovium may be an ideal source of cells for in vitro meniscal fibrocartilage formation, however, favorable in vitro culture conditions for synovium must be established in order to achieve this goal. The objective of this study was to determine cellularity, cell distribution, and extracellular matrix (ECM) formation of equine fibroblast-like synoviocytes (FLS) cultured on synthetic scaffolds, for potential application in synovium-based meniscal tissue engineering. Scaffolds included open-cell poly-L-lactic acid (OPLA) sponges and polyglycolic acid (PGA) scaffolds cultured in static and dynamic culture conditions, and PGA scaffolds coated in poly-L-lactic (PLLA) in dynamic culture conditions.Materials and Methods. Equine FLS were seeded on OPLA and PGA scaffolds, and cultured in a static environment or in a rotating bioreactor for 12 days. Equine FLS were also seeded on PGA scaffolds coated in 2% or 4% PLLA and cultured in a rotating bioreactor for 14 and 21 days. Three scaffolds from each group were fixed, sectioned and stained with Masson’s Trichrome, Safranin-O, and Hematoxylin and Eosin, and cell numbers and distribution were analyzed using computer image analysis. Three PGA and OPLA scaffolds from each culture condition were also analyzed for extracellular matrix (ECM) production via dimethylmethylene blue (sulfated glycosaminoglycan) assay and hydroxyproline (collagen) assay. PLLA coated PGA scaffolds were analyzed using double stranded DNA quantification as areflection of cellularity and confocal laser microscopy in a fluorescent cell viability assay.Results. The highest cellularity occurred in PGA constructs cultured in a rotating bioreactor, which also had a mean sulfated glycosaminoglycan content of 22.3 µg per scaffold. PGA constructs cultured in static conditions had the lowest cellularity. Cells had difficulty adhering to OPLA and the PLLA coating of PGA scaffolds; cellularity was inversely proportional to the concentration of PLLA used. PLLA coating did not prevent dissolution of the PGA scaffolds. All cell scaffold types and culture conditions produced non-uniform cellular distribution.Discussion/Conclusion. FLS-seeding of PGA scaffolds cultured in a rotating bioreactor resulted in the most optimal cell and matrix characteristics seen in this study. Cells grew only in the pores of the OPLA sponge, and could not adhere to the PLLA coating of PGA scaffold, due to the hydrophobic property of PLA. While PGA culture in a bioreactor produced measureable GAG, no culture technique produced visible collagen. For this reason, and due to the dissolution of PGA scaffolds, the culture conditions and scaffolds described here are not recommended for inducing fibrochondrogenesis in equine FLS for meniscal tissue engineering

Report from the STEM 2026 Workshop on Assessment, Evaluation, and Accreditation

A gathering of science, technology, engineering, and math (STEM) higher education stakeholders met in November 2018 to consider the relationship between innovation in education and assessment. When we talk about assessment in higher education, it is inextricably linked to both evaluation and accreditation, so all three were considered. The first question we asked was can we build a nation of learners? This starts with considering the student, first and foremost. As educators, this is a foundation of our exploration and makes our values transparent. As educators, how do we know we are having an impact? As members and implementers of institutions, programs and professional societies, how do we know students are learning and that what they are learning has value? The focus of this conversation was on undergraduate learning, although we acknowledge that the topic is closely tied to successful primary and secondary learning as well as graduate education. Within the realm of undergraduate education, students can experience four-year institutions and two-year institutions, with many students learning at both at different times. Thirty-seven participants spent two days considering cases of innovation in STEM education, learning about the best practices in assessment, and then discussing the relationship of innovation and assessment at multiple levels within the context of higher education. Six working groups looked at course-level, program-level, and institution-level assessment, as well as cross-disciplinary programs, large-scale policy issues, and the difficult-to-name “non-content/cross-content” group that looked at assessment of transferable skills and attributes like professional skills, scientific thinking, mindset, and identity, all of which are related to post-baccalaureate success. These conversations addressed issues that cut across multiple levels, disciplines, and course topics, or are otherwise seen as tangential or perpendicular to perhaps “required” assessment at institutional, programmatic, or course levels. This report presents the context, recommendations, and “wicked” challenges from the meeting participants and their working groups. Along with the recommendations of workshop participants, these intricate challenges weave a complex web of issues that collectively need to be addressed by our community. They generated a great deal of interest and engagement from workshop participants, and act as a call to continue these conversations and seek answers that will improve STEM education through innovation and improved assessment. This material is based upon work supported by the National Science Foundation under Grant No. DUE-1843775. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation

Gene Profiling of Aortic Valve Interstitial Cells under Elevated Pressure Conditions: Modulation of Inflammatory Gene Networks

The study aimed to identify mechanosensitive pathways and gene networks that are stimulated by elevated cyclic pressure in aortic valve interstitial cells (VICs) and lead to detrimental tissue remodeling and/or pathogenesis. Porcine aortic valve leaflets were exposed to cyclic pressures of 80 or 120 mmHg, corresponding to diastolic transvalvular pressure in normal and hypertensive conditions, respectively. Linear, two-cycle amplification of total RNA, followed by microarray was performed for transcriptome analysis (with qRT-PCR validation). A combination of systems biology modeling and pathway analysis identified novel genes and molecular mechanisms underlying the biological response of VICs to elevated pressure. 56 gene transcripts related to inflammatory response mechanisms were differentially expressed. TNF-α, IL-1α, and IL-1β were key cytokines identified from the gene network model. Also of interest was the discovery that pentraxin 3 (PTX3) was significantly upregulated under elevated pressure conditions (41-fold change). In conclusion, a gene network model showing differentially expressed inflammatory genes and their interactions in VICs exposed to elevated pressure has been developed. This system overview has detected key molecules that could be targeted for pharmacotherapy of aortic stenosis in hypertensive patients