Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors

Production of human mesenchymal stem cells for allogeneic cell therapies requires scalable, cost-effective manufacturing processes. Microcarriers enable the culture of anchorage-dependent cells in stirred-tank bioreactors. However, no robust, transferable methodology for microcarrier selection exists, with studies providing little or no reason explaining why a microcarrier was employed. We systematically evaluated 13 microcarriers for human bone marrow-derived MSC (hBM-MSCs) expansion from three donors to establish a reproducible and transferable methodology for microcarrier selection. Monolayer studies demonstrated input cell line variability with respect to growth kinetics and metabolite flux. HBM-MSC1 underwent more cumulative population doublings over three passages in comparison to hBM-MSC2 and hBM-MSC3. In 100 mL spinner flasks, agitated conditions were significantly better than static conditions, irrespective of donor, and relative microcarrier performance was identical where the same microcarriers outperformed others with respect to growth kinetics and metabolite flux. Relative growth kinetics between donor cells on the microcarriers were the same as the monolayer study. Plastic microcarriers were selected as the optimal microcarrier for hBM-MSC expansion. HBM-MSCs were successfully harvested and characterised, demonstrating hBM-MSC immunophenotype and differentiation capacity. This approach provides a systematic method for microcarrier selection, and the findings identify potentially significant bioprocessing implications for microcarrier-based allogeneic cell therapy manufacture. Large-scale production of human bone-marrow derived mesenchymal stem cells (hBM-MSCs) requires expansion on microcarriers in agitated systems. This study demonstrates the importance of microcarrier selection and presents a systematic methodology for selection of an optimal microcarrier. The study also highlights the impact of an agitated culture environment in comparison to a static system, resulting in a significantly higher hBM-MSC yield under agitated conditions.

The impact of information technology on organizations: A study of enterprise resource planning system influences on job design and organizational culture

The primary purpose of this research is to study the linkage between perceived job design characteristics and information system environment characteristics before and after the replacement of a legacy information system with a new type of information system (referred to as an Enterprise Resource Planning or ERP system). A public state University implementing an academic version of an ERP system was selected for the study. Three survey instruments were used to examine the perception of the information system, the job characteristics, and the organizational culture before and after the system implementation. The research participants included two large departments resulting in a sample of 130 workers. Research questions were analyzed using multivariate procedures including factor analysis, path analysis, step-wise regression, and matched pair analysis. ^ Results indicated that the ERP system has introduced new elements into the working environment that has changed the perception of how the job design characteristics and organization culture dimensions are viewed by the workers. The understanding of how the perceived system characteristics align with an individual's perceived job design characteristics is supported by each of the system characteristics significantly correlated in the proposed direction. The stronger support of this relationship becomes visible in the causal flow of the effects seen in the path diagram and in the step-wise regression. The perceived job design characteristics aligning with dimensions of organizational culture are not as strong as the literature suggests. Although there are significant correlations between the job and culture variables, only one relationship can be seen in the causal flow. ^ This research has demonstrated that system characteristics of ERP do contribute to the perception of change in an organization and do support organizational culture behaviors and job characteristics. ^

Responses of neotropical mangrove seedlings grown in monoculture and mixed culture under treatments of hydroperiod and salinity

We investigated the combined effects of salinity and hydroperiod on seedlings of Rhizophora mangle and Laguncularia racemosa grown under experimental conditions of monoculture and mixed culture by using a simulated tidal system. The objective was to test hypotheses relative to species interactions to either tidal or permanent flooding at salinities of 10 or 40 g/l. Four-month-old seedlings were experimentally manipulated under these environmental conditions in two types of species interactions: (1) seedlings of the same species were grown separately in containers from September 2000 to August 2001 to evaluate intraspecific response and (2) seedlings of each species were mixed in containers to evaluate interspecific, competitive responses from August 2002 to April 2003. Overall, L. racemosa was strongly sensitive to treatment combinations while R. mangle showed little effect. Most plant responses of L. racemosa were affected by both salinity and hydroperiod, with hydroperiod inducing more effects than salinity. Compared to R. mangle, L. racemosa in all treatment combinations had higher relative growth rate, leaf area ratio, specific leaf area, stem elongation, total length of branches, net primary production, and stem height. Rhizophora mangle had higher biomass allocation to roots. Species growth differentiation was more pronounced at low salinity, with few species differences at high salinity under permanent flooding. These results suggest that under low to mild stress by hydroperiod and salinity, L. racemosa exhibits responses that favor its competitive dominance over R. mangle. This advantage, however, is strongly reduced as stress from salinity and hydroperiod increase.

The relationships between elementary school principals' emotional intelligence, leadership style and school culture

The role of the principal in school settings and the principal's perceived effect on student achievement have frequently been considered vital factors in school reform. The relationships between emotional intelligence, leadership style and school culture have been widely studied. The literature reveals agreement among scholars regarding the principal's vital role in developing and fostering a positive school culture. The purpose of this study was to explore the relationships between elementary school principals' emotional intelligence, leadership style and school culture. ^ The researcher implemented a non-experimental ex post facto research design to investigate four specific research hypotheses. Utilizing the Qualtrics Survey Software, 57 elementary school principals within a large urban school district in southeast Florida completed the Emotional Quotient Inventory (EQ-i), and 850 of their faculty members completed the Multifactor Leadership Questionnaire (MLQ Form 5X). Faculty responses to the school district's School Climate Survey retrieved from the district's web site were used as the measure of school culture. ^ Linear regression analyses revealed significant positive associations between emotional intelligence and the following leadership measures: Idealized Influence-Attributes (β = .23, p = < .05), Idealized Influence-Behaviors (β = .34, p = < .01), Inspirational Motivation (β = .39, p = < .01) and Contingent Reward (β = .33, p = < .01). Hierarchical regression analyses revealed positive associations between school culture and both transformational and transactional leadership measures, and negative associations between school culture and passive-avoidant leadership measures. Significant positive associations were found between school culture and the principals' emotional intelligence over and above leadership style. Hierarchical linear regressions to test the statistical hypothesis developed to account for alternative explanations revealed significant associations between leadership style and school culture over and above school grade. ^ These results suggest that emotional intelligence merits consideration in the development of leadership theory. Practical implications include suggestions that principals employ both transformational and transactional leadership strategies, and focus on developing their level of emotional intelligence. The associations between emotional intelligence, transformational leadership, Contingent Reward and school culture found in this study validate the role of the principal as the leader of school reform.^

Woven cultures : New insights into Pictish and Viking culture contact using the implements of textile production

This paper is based on the undergraduate dissertation of Lindsey Stirling, which was supervised by Karen Milek, and which won the Society of Medieval Archaeology's John Hurst Memorial Dissertation Prize Acknowledgements This research would not have been possible without the assistance of Dr Martin Goldberg at the National Museum of Scotland, Lynda Aiano at Tankerness House Museum, Orkney, and Beverley Ballin Smith. The authors also wish to thank the two anonymous reviewers who provided valuable comments on earlier versions of this paper.

Woven cultures : New insights into Pictish and Viking culture contact using the implements of textile production

This paper is based on the undergraduate dissertation of Lindsey Stirling, which was supervised by Karen Milek, and which won the Society of Medieval Archaeology's John Hurst Memorial Dissertation Prize Acknowledgements This research would not have been possible without the assistance of Dr Martin Goldberg at the National Museum of Scotland, Lynda Aiano at Tankerness House Museum, Orkney, and Beverley Ballin Smith. The authors also wish to thank the two anonymous reviewers who provided valuable comments on earlier versions of this paper.

The Effects of Brominated Flame Retardants on Thyroid Hormone Homeostasis in Human Placenta Tissues and Cell Culture

Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants (BFRs) that have been heavily used in consumer products such as furniture foams, plastics, and textiles since the mid-1970’s. BFRs are added to products in order to meet state flammability standards intended to increase indoor safety in the event of a fire. The three commercial PBDE mixtures, Penta-, Octa-, and DecaBDE, have all been banned in the United States, however, limited use of DecaBDE is still permitted. PBDEs were phased out of production and added to the Stockholm Convention due to concerns over their environmental persistence and toxicity. Human exposure to PBDEs occurs primarily through the inadvertent ingestion of contaminated house dust, as well as though dietary sources. Despite the phase-out and discontinued use of PBDEs, human exposure to this class of chemicals is likely to continue for decades due to the continued use of treated products and existing environmental reservoirs of PBDEs. Extensive research over the years has shown that PBDEs disrupt thyroid hormone (TH) levels and neurodevelopmental endpoints in rodent and fish models. Additionally, there is growing epidemiological evidence linking PBDE exposure in humans to altered TH homeostasis and neurodevelopmental impairments in children. Due to the importance of THs throughout gestation, there is a great need to understand the effects of BFRs on the developing fetus. Specifically, the placenta plays a critical role in the transport, metabolism, and delivery of THs to the fetal compartment during pregnancy and is a likely target for BFR bioaccumulation and endocrine disruption. The central hypothesis of this dissertation research is that BFRs disrupt the activity of TH sulfotransferase (SULT) enzymes, thereby altering TH concentrations in the placenta.

In the first aim of this dissertation research, the concentrations of PBDEs and 2,4,6-TBP were measured in a cohort of 102 placenta tissue samples from an ongoing pregnancy cohort in Durham, NC. Methods were developed for the extraction and analysis of the BFR analytes. It was found that 2,4,6-TBP was significantly correlated with all PBDE analytes, indicating that 2,4,6-TBP may share common product applications with PBDEs or that 2,4,6-TBP is a metabolite of PBDE compounds. Additionally, this was the first study to measure 2,4,6-TBP in human placenta tissues.

In the second aim of this dissertation research, the placenta tissue concentrations of THs, as well as the endogenous activity of deiodinase (DI) and TH SULT enzymes were quantified using the same cohort of 102 placenta tissue samples. Enzyme activity was detected in all samples and this was the first study to measure TH DI and SULT activity in human placenta tissues. Enzyme activities and TH concentrations were compared with BFR concentrations measured in Aim 1. There were few statistically significant associations observed for the combined data, however, upon stratifying the data set based on infant sex, additional significant associations were observed. For example, among males, those with the highest concentrations of BDE-99 in placenta had T3 levels 0.80 times those with the lowest concentration of BDE-99 (95% confidence interval (CI): 0.59, 1.07). Whereas females with the highest concentrations of BDE-99 in placenta had T3 levels 1.50 times those with the lowest concentration of BDE-99 (95% CI: 1.10, 2.04). Additionally, all BFR analyte concentrations were higher in the placenta of males versus females and they were significantly higher for 2,4,6-TBP and BDE-209. 3,3’-T2 SULT activity was significantly higher in female placenta tissues, while type 3 DI activity was significantly higher in male placenta tissues. This research is the first to show sex-specific differences in the bioaccumulation of BFRs in human placenta tissue, as well as differences in TH concentrations and endogenous DI and SULT activity. The underlying mechanisms of these observed sex differences warrant further investigation.

In the third aim of this dissertation research, the effects of BFRs were examined in a human choriocarcinoma placenta cell line, BeWo. Michaelis-Menten parameters and inhibition curves were calculated for 2,4,6-TBP, 3-OH BDE-47, and 6-OH BDE-47. 2,4,6-TBP was shown to be the most potent inhibitor of 3,3’-T2 SULT activity with a calculated IC50 value of 11.6 nM. It was also shown that 2,4,6-TBP and 3-OH BDE-47 exhibit mixed inhibition of 3,3’-T2 sulfation in BeWo cell homogenates. Next, a series of cell culture exposure experiments were performed using 1, 6, 12, and 24 hour exposure durations. Once again, 2,4,6-TBP was shown to be the most potent inhibitor of basal 3,3’-T2 SULT activity by significantly decreasing activity at the high and medium dose (1 M and 0.5 M, respectively) at all measured time points. Interestingly, BDE-99 was also shown to inhibit basal 3,3’-T2 SULT activity in BeWo cells following the 24 hour exposure, despite exhibiting no inhibitory effects in the BeWo cell homogenate experiments. This indicates that BDE-99 must act through a pathway other than direct enzyme inhibition. Following exposures, the TH concentrations in the cell culture growth media and mRNA expression of TH-related genes were also examined. There was no observed effect of BFR treatment on these endpoints. Future work should focus on determining the downstream biological effects of TH SULT disruption in placental cells, as well as the underlying mechanisms of action responsible for reductions in basal TH SULT activity following BFR exposure.

This was one of the first studies to measure BFRs in a cohort of placenta tissue samples from the United States and the first study to measure THs, DI activity, and SULT activity in human placenta tissues. This research provides a novel contribution to our growing understanding of the effects of BFRs on TH homeostasis within the human placenta, and provides further evidence for sex-specific differences within this important organ. Future research should continue to investigate the effects of environmental contaminants on TH homeostasis within the placenta, as this represents the most critical and vulnerable stage of human development.

Meta-cinematic cultism: between high and low culture

ABSTRACT - Meta-cinema can depict film viewers’ attitudes towards cinema and their type of devotion for films. One subcategory of viewers - which I call meta-spectators - is highly specialized in its type of consumption, bordering on obsession. I contend that there are two main varieties of meta-cinematic reception, not altogether incompatible with one another, despite their apparent differences. As both of them are depicted on meta-cinematic products, the films themselves are the best evidence of my typology. My categories of film viewers are the ‘cinephile’, an elite prone to artistic militancy and the adoration of filmic masters; and the ‘fan’, a low culture consumer keen on certain filmic universes and their respective figures and motifs. I will base my rationale on four films that portray such reception practices: Travelling Avant (Jean-Charles Tacchella, 1987, FRA), The Dreamers (Bernardo Bertolucci, 2003, UK/FRA/ITA); Free Enterprise (Robert Mayer Burnett, 1998, USA); Fanboys (2009, Kyle Newman, USA).

Symbiotic Cities: Buenos Aires, Montevideo, and Mass Culture, 1910-1960

This dissertation examines how Buenos Aires emerged as a creative capital of mass culture and cultural industries in South America during a period when Argentine theater and cinema expanded rapidly, winning over a regional marketplace swelled by transatlantic immigration, urbanization and industrialization. I argue that mass culture across the River Plate developed from a singular dynamic of exchange and competition between Buenos Aires and neighboring Montevideo. The study focuses on the Argentine, Uruguayan, and international performers, playwrights, producers, cultural impresarios, critics, and consumers who collectively built regional cultural industries. The cultural industries in this region blossomed in the interwar period as the advent of new technologies like sound film created profitable opportunities for mass cultural production and new careers for countless theater professionals. Buenos Aires also became a global cultural capital in the wider Hispanic Atlantic world, as its commercial culture served a region composed largely of immigrants and their descendants. From the 1920s through the 1940s, Montevideo maintained a subordinate but symbiotic relationship with Buenos Aires. The two cities shared interlinked cultural marketplaces that attracted performers and directors from the Atlantic world to work in theatre and film productions, especially in times of political upheaval such as the Spanish Civil War and the Perón era in Argentina. As a result of this transnational process, Argentine mass culture became widely consumed throughout South America, competing successfully with Hollywood, European, and other Latin American cinemas and helping transform Buenos Aires into a cosmopolitan metropolis. By examining the relationship between regional and national frames of cultural production, my dissertation contributes to the fields of Latin American studies and urban history while seeking to de-center the United States and Europe from the central framing of transnational history.

Adapting to change in contemporary Irish and Scottish culture: fiction to film

Abstract available: p. [ii]-[iii].

THE TRAITS OF EARLY—BRONZE PONTIC CULTURES IN THE DEVELOPMENT OF OLD UPLAND CORDED WARE (MALOPOLSKA GROUPS) AND ZLOTA CULTURE COMMUNITIES

This publication constitutes the fruits of National Science Centre research projects (grant no 2011/01/M/HS3/02142 – 6 articles) and the National Programme for the Development of the Humanities (grant no 0108/NPH3/H12/82/2014 – 3 articles). We would like to acknowledge and at the same time express our sincere gratitude for the generosity shown by the following at the Adam Mickiewicz University in making this publication possible: the Dean of the Department of History, Institute of Pre-history and the Eastern Institute.

ENEOLITHIC, YAMNAYA, CATACOMB AND BABYNO CULTURE CEMETERIES, PIDLISIVKA, BARROW 1, YAMPIL REGION, VINNITSA OBLAST: ARCHAEOMETRY, CHRONOMETRY AND TAXONOMY

The paper presents excavation results and analytical studies concerning the taxonomic classification of a funerary site identified with the communities of the early ‘barrow cultures’ settling the north-western Black Sea Coast in the 4th/3rd-2nd millennium BC. The study focuses on the ceremonial centres of the Eneolithic, Yamnaya, Catacomb and Babyno cultures.

ENEOLITHIC, YAMNAYA AND NOUA CULTURE CEMETERIES FROM THE FIRST HALF OF THE 3RD AND THE MIDDLE OF THE 2ND MILLENNIUM BC, POROHY, SITE 3A, YAMPIL REGION, VINNITSA OBLAST: ARCHAEOMETRIC AND CHRONOMETRIC DESCRIPTION, RITUAL AND TAXONOMIC-TOPOGENETIC IDENTIFICATION

The paper presents the results of excavations and analytical studies regarding the taxonomic classification of a funeral site associated with the societies of ‘barrow cultures’ of the north-western Black Sea Coast in the first half of the 3rd and the middle of the 2nd millennium BC. The study discusses the ceremonial centres of the Eneolithic, Yamnaya and Noua cultures.