Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrier process

Human mesenchymal stem cell (hMSC) therapies are currently progressing through clinical development, driving the need for consistent, and cost effective manufacturing processes to meet the lot-sizes required for commercial production. The use of animal-derived serum is common in hMSC culture but has many drawbacks such as limited supply, lot-to-lot variability, increased regulatory burden, possibility of pathogen transmission, and reduced scope for process optimization. These constraints may impact the development of a consistent large-scale process and therefore must be addressed. The aim of this work was therefore to run a pilot study in the systematic development of serum-free hMSC manufacturing process. Human bone-marrow derived hMSCs were expanded on fibronectin-coated, non-porous plastic microcarriers in 100mL stirred spinner flasks at a density of 3×10⁵cells.mL^-1 in serum-free medium. The hMSCs were successfully harvested by our recently-developed technique using animal-free enzymatic cell detachment accompanied by agitation followed by filtration to separate the hMSCs from microcarriers, with a post-harvest viability of 99.63±0.03%. The hMSCs were found to be in accordance with the ISCT characterization criteria and maintained hMSC outgrowth and colony-forming potential. The hMSCs were held in suspension post-harvest to simulate a typical pooling time for a scaled expansion process and cryopreserved in a serum-free vehicle solution using a controlled-rate freezing process. Post-thaw viability was 75.8±1.4% with a similar 3h attachment efficiency also observed, indicating successful hMSC recovery, and attachment. This approach therefore demonstrates that once an hMSC line and appropriate medium have been selected for production, multiple unit operations can be integrated to generate an animal component-free hMSC production process from expansion through to cryopreservation.

One's own country and familiar places in the mind's eye:different topological representations for navigational and non-navigational contents

Visual mental imagery is a process that draws on different cognitive abilities and is affected by the contents of mental images. Several studies have demonstrated that different brain areas subtend the mental imagery of navigational and non-navigational contents. Here, we set out to determine whether there are distinct representations for navigational and geographical images. Specifically, we used a Spatial Compatibility Task (SCT) to assess the mental representation of a familiar navigational space (the campus), a familiar geographical space (the map of Italy) and familiar objects (the clock). Twenty-one participants judged whether the vertical or the horizontal arrangement of items was correct. We found that distinct representational strategies were preferred to solve different categories on the SCT, namely, the horizontal perspective for the campus and the vertical perspective for the clock and the map of Italy. Furthermore, we found significant effects due to individual differences in the vividness of mental images and in preferences for verbal versus visual strategies, which selectively affect the contents of mental images. Our results suggest that imagining a familiar navigational space is somewhat different from imagining a familiar geographical space. © 2014 Elsevier Ireland Ltd.

A Note on the Asymptotic Behaviour of a Periodic Multitype Galton-Watson Branching Process

2000 Mathematics Subject Classification: 60J80.

Enhanced process verification through re-engineering the manufacturing procedure

In today’s modern manufacturing industry there is an increasing need to improve internal processes to meet diverse client needs. Process re-engineering is an important activity that is well understood by industry but its rate of application within small to medium size enterprises (SME) is less developed. Business pressures shift the focus of SMEs toward winning new projects and contracts rather than developing long-term, sustainable manufacturing processes. Variations in manufacturing processes are inevitable, but the amount of non-conformity often exceeds the acceptable levels. This paper is focused on the re-engineering of the manufacturing and verification procedure for discrete parts production with the aim of enhancing process control and product verification. The ideologies of the ‘Push’ and ‘Pull’ approaches to manufacturing are useful in the context of process re-engineering for data improvement. Currently information is pulled from the market and prominent customers, and manufacturing companies always try to make the right product, by following customer procedures that attempt to verify against specifications. This approach can result in significant quality control challenges. The aim of this paper is to highlight the importance of process re-engineering in product verification in SMEs. Leadership, culture, ownership and process management are among the main attributes required for the successful deployment of process re-engineering. This paper presents the findings from a case study showcasing the application of a modified re-engingeering method for the manufacturing and verification process. The findings from the case study indicate there are several advantages to implementing the re-engineering method outlined in this paper.

Weak Convergence to the Tangent Process of the Linear Multifractional Stable Motion

2000 Mathematics Subject Classification: 60G18, 60E07

Lp Microlocal Supercritical Markov Branching Processes with Non-Homogeneous Poisson Immigration

2010 Mathematics Subject Classification: 60J80.

Limit Theorems for Non-Critical Branching Processes with Continuous State Space

2000 Mathematics Subject Classification: Primary 60J80, Secondary 60G99.

The non-violence of love:a Hildebrand-Girard encounter

If love is a social as well as a personal reality, it could be fruitful to compare Von Hildebrand’s understanding of love and desire with that of cultural anthropologist René Girard. Girard depicts love and desire as a triangular process which arises from imitation, rather than the result of auto-generative affection. In this sense, Girardian theory would seem to convict Von Hildebrand of what is called the romantic lie wherein desire is thought to arise through the mutual appreciation of two subjects. However, in The Nature of Love Von Hildebrand shows his awareness of the possibility that love can be awakened by imitation. Moreover, the lack of a sufficient reason in Girardian theory for avoiding violence can be answered by turning to Von Hildebrand’s appreciation of the ontological basis for desire.

Learner support and discovery in a virtual non-judgmental environment

This thesis is a qualitative case study that draws upon a grounded genre analysis approach situated within the social constructivist paradigm. The study describes the various obligatory, desired, and optional moves used by post-graduate students as they interacted within an online, non-judgmental environment in order to seek solutions to issues they were experiencing with their research projects or teaching. The postgraduate students or case participants met individually online with me at pre-arranged times to take part in Instant Messenger Cooperative Development (IMCD) (Boon, 2005) 30-minute to one hour sessions via the text-chat function of Skype. Participants took on the role of ‘Explorer’ in order to articulate their thoughts and ideas about their research. I took on the role of ‘Understander’ to provide support to each Explorer by reflecting my understanding of the ongoing articulations as the Explorers investigated their specific issues, determined possible ways to overcome them, made new discoveries, and formulated plans of action regarding the best way for them to move forward. The description of generic moves covers 32 IMCD sessions collected over a threeyear period (2009-2012) from 10 different participants (A-J). Data collected is drawn from live IMCD sessions, field notes, and post-session email feedback from participants. In particular, the thesis focuses on describing the specific generic moves of Explorers within IMCD sessions as they seek satisfactory resolutions to particular research or pedagogic puzzles. It also provides a detailed description of a longitudinal case (Participant A – four sessions), a one-session case (Participant B – one session), and an outlier case in which the Explorer underwent a negative IMCD experience. The thesis concludes by arguing that IMCD is a highly effective tool that helps facilitate the research process for both distance-learning and on-campus students and has the potential to be utilized across all disciplines at the tertiary level.

Optical fibre limits:an approach using ASE channel estimation

In this talk we investigate the usage of spectrally shaped amplified spontaneous emission (ASE) in order to emulate highly dispersed wavelength division multiplexed (WDM) signals in an optical transmission system. Such a technique offers various simplifications to large scale WDM experiments. Not only does it offer a reduction in transmitter complexity, removing the need for multiple source lasers, it potentially reduces the test and measurement complexity by requiring only the centre channel of a WDM system to be measured in order to estimate WDM worst case performance. The use of ASE as a test and measurement tool is well established in optical communication systems and several measurement techniques will be discussed [1, 2]. One of the most prevalent uses of ASE is in the measurement of receiver sensitivity where ASE is introduced in order to degrade the optical signal to noise ratio (OSNR) and measure the resulting bit error rate (BER) at the receiver. From an analytical point of view noise has been used to emulate system performance, the Gaussian Noise model is used as an estimate of highly dispersed signals and has had consider- able interest [3]. The work to be presented here extends the use of ASE by using it as a metric to emulate highly dispersed WDM signals and in the process reduce WDM transmitter complexity and receiver measurement time in a lab environment. Results thus far have indicated [2] that such a transmitter configuration is consistent with an AWGN model for transmission, with modulation format complexity and nonlinearities playing a key role in estimating the performance of systems utilising the ASE channel emulation technique. We conclude this work by investigating techniques capable of characterising the nonlinear and damage limits of optical fibres and the resultant information capacity limits. REFERENCES McCarthy, M. E., N. Mac Suibhne, S. T. Le, P. Harper, and A. D. Ellis, “High spectral efficiency transmission emulation for non-linear transmission performance estimation for high order modulation formats," 2014 European Conference on IEEE Optical Communication (ECOC), 2014. 2. Ellis, A., N. Mac Suibhne, F. Gunning, and S. Sygletos, “Expressions for the nonlinear trans- mission performance of multi-mode optical fiber," Opt. Express, Vol. 21, 22834{22846, 2013. Vacondio, F., O. Rival, C. Simonneau, E. Grellier, A. Bononi, L. Lorcy, J. Antona, and S. Bigo, “On nonlinear distortions of highly dispersive optical coherent systems," Opt. Express, Vol. 20, 1022-1032, 2012.

Immunological synapse: a mathematical model of the bond formation process:mathematical modelling of the immature synapse

The cell:cell bond between an immune cell and an antigen presenting cell is a necessary event in the activation of the adaptive immune response. At the juncture between the cells, cell surface molecules on the opposing cells form non-covalent bonds and a distinct patterning is observed that is termed the immunological synapse. An important binding molecule in the synapse is the T-cell receptor (TCR), that is responsible for antigen recognition through its binding with a major-histocompatibility complex with bound peptide (pMHC). This bond leads to intracellular signalling events that culminate in the activation of the T-cell, and ultimately leads to the expression of the immune eector function. The temporal analysis of the TCR bonds during the formation of the immunological synapse presents a problem to biologists, due to the spatio-temporal scales (nanometers and picoseconds) that compare with experimental uncertainty limits. In this study, a linear stochastic model, derived from a nonlinear model of the synapse, is used to analyse the temporal dynamics of the bond attachments for the TCR. Mathematical analysis and numerical methods are employed to analyse the qualitative dynamics of the nonequilibrium membrane dynamics, with the specic aim of calculating the average persistence time for the TCR:pMHC bond. A single-threshold method, that has been previously used to successfully calculate the TCR:pMHC contact path sizes in the synapse, is applied to produce results for the average contact times of the TCR:pMHC bonds. This method is extended through the development of a two-threshold method, that produces results suggesting the average time persistence for the TCR:pMHC bond is in the order of 2-4 seconds, values that agree with experimental evidence for TCR signalling. The study reveals two distinct scaling regimes in the time persistent survival probability density prole of these bonds, one dominated by thermal uctuations and the other associated with the TCR signalling. Analysis of the thermal fluctuation regime reveals a minimal contribution to the average time persistence calculation, that has an important biological implication when comparing the probabilistic models to experimental evidence. In cases where only a few statistics can be gathered from experimental conditions, the results are unlikely to match the probabilistic predictions. The results also identify a rescaling relationship between the thermal noise and the bond length, suggesting a recalibration of the experimental conditions, to adhere to this scaling relationship, will enable biologists to identify the start of the signalling regime for previously unobserved receptor:ligand bonds. Also, the regime associated with TCR signalling exhibits a universal decay rate for the persistence probability, that is independent of the bond length.

A hazai védekezőtől a regionális „támadóig” – az OTP nemzetköziesedésének időbeni alakulása (From the domestic defender to the regional striker – a timebased process of the OTP Bank’s internationalization)

A vállalatok nemzetköziesedése egy széles körben vizsgált szakterület, különösképpen olyan vállalatok esetében (az ún. tradicionális multik), amelyek hagyományosan nemzetközi orientációjú országokból (Triád) származnak. A nemzetközi piacokon viszonylag újnak számító közép-kelet-európai cégek terjeszkedése az üzleti kontextus változásával párhuzamosan zajlott. Következésképpen ahhoz, hogy megértsük a nemzetköziesedés időben zajló folyamatát, a vállalatok nemzetközi evolúcióját a kontextussal összefüggésben kell vizsgálni. A vállalatok nemzetköziesedésének legelterjedtebb kutatási módszerei a nagymintás keresztmetszeti vizsgálatok, amelyek nem adnak lehetőséget a nemzetközi fejlődés időbeni alakulásának megfigyelésére és a mintázat mögött rejlő valós okok feltárására. A jelen tanulmány egy longitudinális kutatási modell mentén vizsgálja egy nem tradicionális multi, a hazai OTP nemzetköziesedésének időbeni alakulását, feltárva a nemzetközivé válás folyamata során kialakuló komplex kapcsolatrendszert. _______ The purpose of the present paper is to describe the internationalisation process of a non-traditional multinational, the Hungarian OTP Bank and to understand the dynamic linkages between the spatial and temporal context and the content of firm internationalization using a phenomenon oriented longitudinal research design. The paper contributes to the time-based processual view of firm internationalization arguing that in order to understand firm internationalization along time one should observe organizations and their contextual environments as complex interacting processes. By analysing the interaction between the multi-level processes that shape internationalization, one can explore the reasons behind the dynamic profile of firm internationalization. The paper applies an interpretative approach focusing on local causality.

Valued occupational roles and life satisfaction among south Florida HIV-infected and non-infected homosexual males: An exploratory study

A study was conducted in order to describe and understand the occupational role behavior of gay men with particular interest on understanding the impact of an HIV diagnosis on their roles and life satisfaction. A 137-item questionnaire was developed for this study and distributed through various gay community groups in south Florida. The process resulted in ranked description of 24 valued occupational roles of gay men (n = 80) along with a general understanding of potential shifts that may occur in those roles as a result of becoming infected with HIV. The study concludes that a diagnosis of HIV infection impacts gay men both by altering their value for particular roles and by darkening the images they hold of their future. The study also identified key factors which may contribute to the greater life satisfaction of gay men including greater confidence in personal knowledge of HIV, a more optimistic outlook an life, the use of stronger coping styles, and perceiving stronger social support and lower stress. ^

The nitrate to ammonia and ceramic (NAC) process for the denitration and immobilization of low-level radioactive liquid waste (LLW)

Hazardous radioactive liquid waste is the legacy of more than 50 years of plutonium production associated with the United States' nuclear weapons program. It is estimated that more than 245,000 tons of nitrate wastes are stored at facilities such as the single-shell tanks (SST) at the Hanford Site in the state of Washington, and the Melton Valley storage tanks at Oak Ridge National Laboratory (ORNL) in Tennessee. In order to develop an innovative, new technology for the destruction and immobilization of nitrate-based radioactive liquid waste, the United State Department of Energy (DOE) initiated the research project which resulted in the technology known as the Nitrate to Ammonia and Ceramic (NAC) process. However, inasmuch as the nitrate anion is highly mobile and difficult to immobilize, especially in relatively porous cement-based grout which has been used to date as a method for the immobilization of liquid waste, it presents a major obstacle to environmental clean-up initiatives. Thus, in an effort to contribute to the existing body of knowledge and enhance the efficacy of the NAC process, this research involved the experimental measurement of the rheological and heat transfer behaviors of the NAC product slurry and the determination of the optimal operating parameters for the continuous NAC chemical reaction process. Test results indicate that the NAC product slurry exhibits a typical non-Newtonian flow behavior. Correlation equations for the slurry's rheological properties and heat transfer rate in a pipe flow have been developed; these should prove valuable in the design of a full-scale NAC processing plant. The 20-percent slurry exhibited a typical dilatant (shear thickening) behavior and was in the turbulent flow regime due to its lower viscosity. The 40-percent slurry exhibited a typical pseudoplastic (shear thinning) behavior and remained in the laminar flow regime throughout its experimental range. The reactions were found to be more efficient in the lower temperature range investigated. With respect to leachability, the experimental final NAC ceramic waste form is comparable to the final product of vitrification, the technology chosen by DOE to treat these wastes. As the NAC process has the potential of reducing the volume of nitrate-based radioactive liquid waste by as much as 70 percent, it not only promises to enhance environmental remediation efforts but also effect substantial cost savings. ^

Comparative analysis of a moderating and mediating model of stress, appraisal, and coping in Hispanic and non-Hispanic Alzheimer's disease caregivers

This dissertation utilizes a cross-sectional study to examine the phenomenon of caregiving within a theoretically grounded stress, appraisal, and coping model. Hispanic and non-Hispanic caregivers were studied to examine the factors associated with variance in caregiver appraisal, coping, and outcomes of caregiving strain (depression and somatic complaints) and caregiving gain (life satisfaction, mastery, and personal gain). A purposive sampling strategy was used to recruit 204 Alzheimer's disease caregivers in South Florida. A self-report questionnaire was used to collect demographic data, and to measure stress, appraisal, coping, and psychological well-being of caregivers. Regression equations were developed to compare moderating and mediating models of appraisal and coping. Emotion-focused coping skills were found to significantly moderate the effects of stress (F [1,195] = 4.62, p < .05), explaining approximately 21% of the variance in satisfaction was found to moderate the effects of stress (F [1,195] = 7.09; p < .05), explaining approximately 27% of the variance in personal gain and approximately 8% of the variance in life satisfaction (F [1,195] = 4.14; p < .05). Appraisal of Burden was found to significantly mediate the effects of stress, explaining approximately 30% of the variance in somatic complaints (F [1,196] = 31.60; p < .001) and 32% of the variance in depression (F [1,196] = 38.18; p < .001). The results of the analyses indicate that appraisal and coping skills are important variables in the stress process. The results of this study underscore the importance of accounting for positive and negative outcomes in providing a fuller understanding of the stress, appraisal and coping process of Alzheimer's Disease caregivers. ^