Developing a scalable model of recombinant protein yield from Pichia pastoris:the influence of culture conditions, biomass and induction regime

Background The optimisation and scale-up of process conditions leading to high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences. Typical experiments rely on varying selected parameters through repeated rounds of trial-and-error optimisation. To rationalise this, several groups have recently adopted the 'design of experiments' (DoE) approach frequently used in industry. Studies have focused on parameters such as medium composition, nutrient feed rates and induction of expression in shake flasks or bioreactors, as well as oxygen transfer rates in micro-well plates. In this study we wanted to generate a predictive model that described small-scale screens and to test its scalability to bioreactors. Results Here we demonstrate how the use of a DoE approach in a multi-well mini-bioreactor permitted the rapid establishment of high yielding production phase conditions that could be transferred to a 7 L bioreactor. Using green fluorescent protein secreted from Pichia pastoris, we derived a predictive model of protein yield as a function of the three most commonly-varied process parameters: temperature, pH and the percentage of dissolved oxygen in the culture medium. Importantly, when yield was normalised to culture volume and density, the model was scalable from mL to L working volumes. By increasing pre-induction biomass accumulation, model-predicted yields were further improved. Yield improvement was most significant, however, on varying the fed-batch induction regime to minimise methanol accumulation so that the productivity of the culture increased throughout the whole induction period. These findings suggest the importance of matching the rate of protein production with the host metabolism. Conclusion We demonstrate how a rational, stepwise approach to recombinant protein production screens can reduce process development time.

Rotational co-culture of clonal β-cells with endothelial cells:effect of PPAR-γ agonism in vitro on insulin and VEGF secretion

Aim: Delayed graft revascularization impedes the success of human islet transplantation. This study utilized rotational co-culture of insulin secreting ß-cells with human umbilical vein endothelial cells (HUVECs) and a peroxisome proliferator-activated receptor gamma (PPAR-?) agonist to promote insulin and vascular endothelial growth factor (VEGF) secretory function. Methods: Clonal BRIN-BD11 (D11) cells were maintained in static culture (SC) and rotational culture (RC) ± HUVEC and ± the TZD (thiazolidinedione) rosiglitazone (10 mmol/l) as a specific PPAR-? agonist. HUVECs were cultured in SC and RC ± D11 and ± TZD. D11 insulin secretion was induced by static incubation with low glucose (1.67 mmol/l), high glucose (16.7 mmol/l) and high glucose with 10 mmol/l theophylline (G+T) and assessed by enzyme-linked immunosorbent assay (ELISA). HUVEC proliferation was determined by ATP luminescence, whereas VEGF secretion was quantified by ELISA. Co-cultured cells were characterized by immunostaining for insulin and CD31. Results: D11 SC and RC showed enhanced insulin secretion in response to 16.7 mmol/l and G+T (p <0.01); without significant alteration by the TZD. Co-culture with HUVEC in SC and RC also increased D11 insulin secretion when challenged with 16.7 mmol/l and G+T (p <0.01), and this was slightly enhanced by the TZD. The presence of HUVEC increased D11 SC and RC insulin secretion in response to high glucose and G+T, respectively (p <0.01). Addition of the TZD increased SC and RC HUVEC ATP content (p <0.01) and VEGF production (p <0.01) in the presence and absence of D11 cells. Conclusions: Rotational co-culture of insulin secreting cells with endothelial cells, and exposure to a PPAR-? agonist may improve the prospects for graft revascularization and function after implantation. © 2011 Blackwell Publishing Ltd.

Performance-based vs socially supportive culture:a cross-national study of descriptive norms and entrepreneurship

This paper is a cross-national study testing a framework relating cultural descriptive norms to entrepreneurship in a sample of 40 nations. Based on data from the Global Leadership and Organizational Behavior Effectiveness project, we identify two higher-order dimensions of culture – socially supportive culture (SSC) and performance-based culture (PBC) – and relate them to entrepreneurship rates and associated supply-side and demand-side variables available from the Global Entrepreneurship Monitor. Findings provide strong support for a social capital/SSC and supply-side variable explanation of entrepreneurship rate. PBC predicts demand-side variables, such as opportunity existence and the quality of formal institutions to support entrepreneurship.

Investigating the effect of geographical distances and cultural proximity on the Hungarian wine trade

Wine is a very special product from an economic, cultural, and sociological point of view. Wine culture and wine trade play an important role in Hungary. The effect of cultural and geographical proximity on international trade has already been proven in the international trade literature. The size of bilateral trade flows between any two countries can be approximated by the gravity theory of trade. The gravity model provides empirical evidence of the relationship between the size of the economies, the distances between them, and their trade. This paper seeks to analyse the effect of cultural and geographical proximity on Hungary’s bilateral wine trade between 2000 and 2012, employing the gravity equation. The analysis is based on data from the World Bank WITS, WDI, as well as CEPII, and WTO databases. I apply OLS, Random Effects, Poisson, Pseudo-Poisson-Maximum-Likelihood and Heckman two stage estimators to calculate the gravity regression. The results show that in the case of Hungary, cultural similarity and trade liberalisation have a positive impact, while geographical distance, landlockedness, and contiguity have a negative impact on Hungarian wine exports.

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. ^

Environment, culture, and medicinal plant knowledge in an indigenous Amazonian community

Diminishing cultural and biological diversity is a current global crisis. Tropical forests and indigenous peoples are adversely affected by social and environmental changes caused by global political and economic systems. The purpose of this thesis was to investigate environmental and livelihood challenges as well as medicinal plant knowledge in a Yagua village in the Peruvian Amazon. Indigenous peoples’ relationships with the environment is an important topic in environmental anthropology, and traditional botanical knowledge is an integral component of ethnobotany. Political ecology provides a useful theoretical perspective for understanding the economic and political dimensions of environmental and social conditions. This research utilized a variety of ethnographic, ethnobotanical, and community-involved methods. Findings include data and analyses about the community’s culture, subsistence and natural resource needs, organizations and institutions, and medicinal plant use. The conclusion discusses the case study in terms of the disciplinary framework and offers suggestions for research and application.

Quantifying the Impacts of the Recent Economic Crisis on Regional Tourism Industry and Economy

The purpose of this study is to explore the accuracy issue of the Input-Output model in quantifying the impacts of the 2007 economic crisis on a local tourism industry and economy. Though the model has been used in the tourism impact analysis, its estimation accuracy is rarely verified empirically. The Metro Orlando area in Florida is investigated as an empirical study, and the negative change in visitor expenditure between 2007 and 2008 is taken as the direct shock. The total impacts are assessed in terms of output and employment, and are compared with the actual data. This study finds that there are surprisingly large discrepancies among the estimated and actual results, and the Input-Output model appears to overestimate the negative impacts. By investigating the local economic activities during the study period, this study made some exploratory efforts in explaining such discrepancies. Theoretical and practical implications are then suggested.

Intra-regional Tourism and Challenges Facing Hong Kong's Hotel Industry

Asia is experiencing a rapid growth in intra-Asian tourism, and is finding that the spending priorities of these new visitor markets is quite different from traditional markets. Not only have Hong Kong's markets changed, but the economic operational environment is becoming increasingly difficult as a result of the change in sovereignty in 1997, increasing land prices, and new regulations. The current structure of the hotel industry is out of balance with the demands of these new markets. Hong Kong now needs to consider some intervention in the hotel industry to further encourage the development of properties in this mid-market.

Authentic Experiences Assessment Instrument: The Case of Millennial Students and Cultural Attractions in Central Florida

Three types of authenticity were chosen to analyze cultural attractions—they were: staged or real authenticity, sensory authenticity, and existential authenticity. A group of millennium students that visited Orlando’s cultural attractions from the University of Central Florida were part of this study. Results show that the term “culture” is unclear among students. Also gender might play a role in determining what is perceived as “authenticity.” In general the focus group discussion had a consensus that these three types of authenticity were the right ones to measured cultural attractions, people, and behaviors.

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.^

Customer Satisfaction and Behavioral Intentions: The Case of Aruba-- Small Island Nation

Tourism studies related to small island destinations have become a research stream amongst many academics in recent years. The current study investigates tourist satisfaction related to a tour operator on the island of Aruba that specializes in jeep and bus tours. As there is an increased expenditure pattern for these types of activities, companies are looking for ways to improve customer satisfaction and behavioral intentions. Results indicate that tourists are generally satisfied with the tour company; however a difference in satisfaction ratings was obtained for respondents 61 years old or above. Four factors were extracted from tourists’ satisfaction attributes and three of these factors, including the overall tour value, tour guide, and sound systems used during these tours, were found to be strong predictors of behavioral intentions measured by the likelihood to rebook and the likelihood to recommend the tour company to family and friends. Operational and marketing strategies were suggested based on the findings of the study.

Tequila at Sunrise: Spring Break Travel Motivations and Risk Behaviors of American Students in Acapulco, Mexico

Little research has been completed regarding spring break motivations and behaviors of American students in foreign destinations, specifically in Mexico. This paper looks at push and pull motivations in relation to drug and alcohol consumption and findings indicate greater drug and alcohol use among those who selected “party reputation” and “to go wild” as travel motivations. Binge drinking, sexual activity, and drug use among students on spring break in Acapulco, Mexico were also analyzed and compared to past findings within the United States. Results suggest that students are involved in heavy alcohol consumption and significant drug use. Additionally, high rates of sexual activity occur on spring break and results suggest low condom use, placing students at higher risks for the contraction of sexually transmitted infections.

Looking At Residents’ Attitudes towards Spring Break Tourism in Texas through the Lens of Community Attachment

The growth of spring break tourism in many destinations has become problematic, predominantly due to the excessive behaviour of college students. This paper examines residents’ attitudes toward spring break tourism in South Padre Island (located in Texas, USA) through the lens of community attachment. By understanding the attitudes of residents of the host communities, tourism planners and policy-makers can create policies to shape the character of tourism according to the residents’ needs. The findings suggest that, at this point in time, community residents perceive that the benefits of spring break tourism benefits exceed its’ costs. Also, the short and intense season of spring break tourism allows residents to better deal with social costs.

Tourist See Tourist Do: The Influence of Hollywood Movies and Television on Tourism Motivation and Activity Behavior

The purpose of this study was to investigate the influence of Hollywood movies and television (movies/TV) on US viewer’s motivation to travel to and participate in activities featured in Hollywood movies/TV productions. A survey was administered in an online format to a convenience sample of 433 respondents via Qualtrics. Factor analysis, correlation, and regression was employed to explore relationships between the variables. Findings identified a profile of Hollywood movies/TV viewers, sources of information used to determine destination choice, and level of involvement among viewers of Hollywood movies/TV productions. Additionally, this study explored the relationships between Hollywood movies/TV productions, tourist motivations, and the propensity to participate in activities featured. Findings indicate that Hollywood movies/TV productions have a positive impact on viewer involvement and that movie/TV related tourism is likely to be affected by movie and TV viewing preference and destination image. The results identify that the predictor “TV viewing behavior” is the strongest predictor of entertainmentmotivated tourism, followed by “destination image” and “movie viewing behavior.” Findings also indicate that “destination image” is the strongest predictor of movie-related activities and that the image portrayed in a movie does influence the viewer’s inclination to visit and participate in activities featured in a movie.

Environment, Culture, and Medicinal Plant Knowledge in an Indigenous Amazonian Community

Diminishing cultural and biological diversity is a current global crisis. Tropical forests and indigenous peoples are adversely affected by social and environmental changes caused by global political and economic systems. The purpose of this thesis was to investigate environmental and livelihood challenges as well as medicinal plant knowledge in a Yagua village in the Peruvian Amazon. Indigenous peoples’ relationships with the environment is an important topic in environmental anthropology, and traditional botanical knowledge is an integral component of ethnobotany. Political ecology provides a useful theoretical perspective for understanding the economic and political dimensions of environmental and social conditions. This research utilized a variety of ethnographic, ethnobotanical, and community-involved methods. Findings include data and analyses about the community’s culture, subsistence and natural resource needs, organizations and institutions, and medicinal plant use. The conclusion discusses the case study in terms of the disciplinary framework and offers suggestions for research and application.