The RGBarrier assay: the parallel study of gene regulatory element performance at defined chromosomal locations

Vertebrate genomes are organised into a variety of nuclear environments and chromatin states that have profound effects on the regulation of gene transcription. This variation presents a major challenge to the expression of transgenes for experimental research, genetic therapies and the production of biopharmaceuticals. The majority of transgenes succumb to transcriptional silencing by their chromosomal environment when they are randomly integrated into the genome, a phenomenon known as chromosomal position effect (CPE). It is not always feasible to target transgene integration to transcriptionally permissive “safe harbour” loci that favour transgene expression, so there remains an unmet need to identify gene regulatory elements that can be added to transgenes which protect them against CPE. Dominant regulatory elements (DREs) with chromatin barrier (or boundary) activity have been shown to protect transgenes from CPE. The HS4 element from the chicken beta-globin locus and the A2UCOE element from a human housekeeping gene locus have been shown to function as DRE barriers in a wide variety of cell types and species. Despite rapid advances in the profiling of transcription factor binding, chromatin states and chromosomal looping interactions, progress towards functionally validating the many candidate barrier elements in vertebrates has been very slow. This is largely due to the lack of a tractable and efficient assay for chromatin barrier activity. In this study, I have developed the RGBarrier assay system to test the chromatin barrier activity of candidate DREs at pre-defined isogenic loci in human cells. The RGBarrier assay consists in a Flp-based RMCE reaction for the integration of an expression construct, carrying candidate DREs, in a pre-characterised chromosomal location. The RGBarrier system involves the tracking of red, green and blue fluorescent proteins by flow cytometry to monitor on-target versus off-target integration and transgene expression. The analysis of the reporter (GFP) expression for several weeks gives a measure of the protective ability of each candidate elements from chromosomal silencing. This assay can be scaled up to test tens of new putative barrier elements in the same chromosomal context in parallel. The defined chromosomal contexts of the RGBarrier assays will allow for detailed mechanistic studies of chromosomal silencing and DRE barrier element action. Understanding these mechanisms will be of paramount importance for the design of specific solutions for overcoming chromosomal silencing in specific transgenic applications.

Communication across cultures? An intercultural approach to customer service in the hotel industry: a study with globally branded hotels in the United Kingdom

In a professional and business-social context such as that of global hotel brands in the United Kingdom, intercultural communication, contacts and relationships are found at the heart of daily operations and of customer service. A large part of the clientele base of hotels in the United Kingdom is formed by individuals who belong to different cultural groups that travel in the country either for leisure or business. At the same time, the global workforce which is recruited in the hotel industry in the United Kingdom is a reality here to stay. Global travelling and labor work mobility are phenomena which have been generated by changes which occur on a socio-economic, cultural and political level due to the phenomenon of globalization. The hotel industry is therefore well acquainted with the essence of different cultures either to be accommodated within hotel premises, as in the case of external customers, or of diversity management where different cultures are recruited in the hotel industry, as in the case of internal customers. This thesis derives from research conducted on eight different global hotel brands in the United Kingdom in particular, with reference to three, four and five star categories. The research aimed to answer the question of how hotels are organized in order to address issues of intercultural communication during customer service and if intercultural barriers arise during the intercultural interaction of hotel staff and global customers. So as to understand how global hotel brands operate the research carried out focused in three main areas relating to each hotel: organizational culture, customer service–customer care and intercultural issues. The study utilized qualitative interviews with hotel management staff and non-management staff from different cultural backgrounds, public space observations between customers and staff during check-in and checkout in the reception area and during dining at the café-bar and restaurant. Thematic analysis was also applied to the official web page of each hotel and to job advertisements to enhance the findings from the interviews and the observations. For the process of analysis of the data interpretive (hermeneutic) phenomenology of Martin Heidegger has been applied. Generally, it was found that hotel staff quite often feel perplexed by how to deal with and how to overcome, for instance, language barriers and religious issues and how to interpret non verbal behaviors or matters on food culture relating to the intercultural aspect of customer service. In addition, it was interesting to find that attention to excellent customer service on the part of hotel staff is a top organizational value and customer care is a priority. Despite that, the participating hotel brands appear to have not yet, realized how intercultural barriers can affect the daily operation of the hotel, the job performance and the psychology of hotel staff. Employees indicated that they were keen to receive diversity training, provided by their organizations, so as to learn about different cultural needs and expand their intercultural skills. The notion of diversity training in global hotel brands is based on the sense that one of the multiple aims of diversity management as a practice and policy in the workplace of hotels is the better understanding of intercultural differences. Therefore global hotel brands can consider diversity training as a practice which will benefit their hotel staff and clientele base at the same time. This can have a distinctive organizational advantage for organizational affairs in the hotel industry, with potential to influence the effectiveness and performance of hotels.