bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reliable method for high quality His-tagged and untaggedE. coliphosphoribosyl phosphate synthase (Prs) purification
Authors: Maria Walter Beata , Aneta Szulc , Monika Glinkowska 
Published: December 2019

ABSTRACTPrs (phosphoribosyl pyrophosphate synthase) is a broadly conserved protein that synthesises 5-phosphoribosyl 1-pyrophospate (PRPP); a substrate for biosynthesis of at least 10 enzymatic pathways including biosynthesis of DNA building blocks – purines and pyrimidines. InEscherichia coli, it is a protein of homo-hexameric quaternary structure, which can be challenging to work with, due to frequent aggregation and activity loss. Several studies showed brief purification protocols for various bacterial PRPP synthases, in most cases involving ammonium sulfate precipitation.Here, we provide a protocol for expression ofE. coliPrs protein in Rosetta (DE3) and BL21 (DE3) pLysE strains and a detailed method for His-Prs and untagged Prs purification on nickel affinity chromatography columns. This protocol allows purification of proteins with high yield, purity and activity. We report here N-terminally His-tagged protein fusions, stable and active, providing that the temperature around 20 °C is maintained at all stages, including centrifugation. Moreover, we successfully applied this method to purify two enzyme variants with K194A and G9S alterations. The K194A mutation in conserved lysine residue results in protein variant unable to synthetize PRPP, while the G9S alteration originates fromprs-2allele variant which was previously related to thermo-sensitive growth. His-PrsG9S protein purified here, exhibited comparable activity as previously observedin-vivosuggesting the proteins purified with our protocol resemble their physiological state.The protocol for Prs purification showed here indicates guidance to improve stability and quality of the protein and to ensure more reliable results in further assaysin-vitro.
Cell shape changes during larval body plan development in Clytia hemisphaerica
Authors: Kraus Yulia , Chevalier Sandra , Houliston Evelyn 
Published: December 2019

AbstractThe cnidarian “planula” larva shows radial symmetry around a polarized, oral-aboral, body axis and comprises two epithelia cell layers, ectodermal and endodermal. This simple body plan is set up during gastrulation, a process which proceeds by a variety of modes amongst the diverse cnidarian species. In the hydrozoan laboratory model Clytia hemisphaerica, gastrulation involves a process termed unipolar cell ingression, in which the endoderm derives from mass ingression of individual cells via a process of epithelial-mesenchymal transition (EMT) around the future oral pole of an epithelial embryo. This contrasts markedly from the gastrulation mode in the anthozoan cnidarian Nematostella vectensis, in which endoderm formation primarily relies on cell sheet invagination. To understand the cellular basis of gastrulation in Clytia we have characterized in detail successive cell morphology changes during planula formation by Scanning and Transmission Electron Microscopy combined with confocal imaging. These changes successively accompany epithialization of the blastoderm, EMT occurring in the oral domain through the bottle cell formation and ingression, cohesive migration and intercalation of ingressed cells with mesenchymal morphology, and their epithelialization to form the endoderm. From our data, we have reconstructed the cascade of morphogenetic events leading to the formation of planula larva. We also matched the domains of cell morphology changes to the expression of selected regulatory and marker genes expressed during gastrulation. We propose that cell ingression in Clytia not only provides the endoderm, but generates internal forces that shape the embryo in the course of gastrulation. These observations help build a more complete understanding of the cellular basis of morphogenesis and of the evolutionary plasticity of cnidarian gastrulation modes.
Mutation of an L-Type Calcium Channel Gene Leads to a Novel Human Primary Cellular Immunodeficiency
Authors: Fenninger Franz , Stanwood Shawna R. , Lu Chieh-Ju , Pfeifer Cheryl G. , Henrickson Sarah E. , Khan Omar , O’Boyle Kaitlin C. , Maurer Kelly , Ruffner Melanie , Herati Ramin S. , Romberg Neil D. , John Wherry E. , Sullivan Kathleen E. , Jefferies Wilfred A. 
Published: December 2019

AbstractHuman primary immunodeficiencies are inherited diseases that can provide valuable insight into the immune system. Calcium (Ca2+) is a vital secondary messenger in T lymphocytes regulating a vast array of important events including maturation, homeostasis, activation, and apoptosis and can enter the cell through CRAC, TRP, and Cav channels. Here we describe three Cav1.4-deficient siblings presenting with X-linked incomplete congenital stationary night blindness as well as an immune phenotype characterized by several recurrent infections. Complete exome sequencing demonstrated that the patients share only a single pathogenic allele; a R625X (p.Arg625Ter) point mutation that leads to a premature stop codon in theCACNA1Fgene encoding the L-type Ca2+channel Cav1.4. The subjects uniformly exhibited an expansion of central and effector memory T lymphocytes, and evidence of T lymphocytes exhaustion with corresponding upregulation of inhibitory receptors. Moreover, the sustained elevated levels of activation markers on B lymphocytes suggest that they are in a chronic state of activation. Finally, the T lymphocytes from patients andCACNA1Fknockdown Jurkat T lymphocytes exhibited a reduced Ca2+flux, compared to controls. This is the first example where the mutation of any Cav channel causes a primary immunodeficiency in humans and establishes the physiological importance of Cav channels in the human immune system.
Relationships and genome evolution of polyploid Salix species revealed by RAD sequencing data
Authors: Wagner Natascha D. , He Li , Hörandl Elvira 
Published: December 2019

AbstractDespite the general progress in using next generation sequencing techniques for evolutionary research questions, the analysis of polyploid species is still hampered by the lack of suitable analytical tools and the statistical difficulties of dealing with more than two alleles per locus. Polyploidization and especially allopolyploidy leads to new combinations of traits by combining genomes of two or more parental species. This enhances the adaptive potential and often results in speciation. However, multiple origins of polyploids, backcrossing to the parental species and post-origin evolution can strongly influence the genome composition of polyploid species. Here, we used RAD sequencing, which revealed 23,393 loci and 320,010 high quality SNPs, to analyze the relationships and origin of seven polyploid species of the diverse genus Salix by utilizing a phylogenomic and a network approach, as well as analyzing the genetic structure and composition of the polyploid genome in comparison to putative parental species. We adapted the SNiPloid pipeline that was originally developed to analyse SNP composition of recently established allotetraploid crop lineages to RAD sequencing data by using concatenated RAD loci as reference. Our results revealed a well-resolved phylogeny of 35 species of Eurasian shrub willows (Salix subg. Chamaetia/Vetrix), including 28 diploid and 7 polyploid species. Polyploidization in willows appears to be predominantly connected to hybridization, i.e. to an allopolyploid origin of species. More ancient allopolyploidization events involving hybridization of more distantly related, ancestral lineages were observed for two hexaploid and one octoploid species. Our data suggested a more recent allopolyploid origin for the included tetraploids within the major subclades and identified putative parental taxa that appear to be plausible in the context of geographical, morphological and ecological patterns. SNiPloid and HyDe analyses disentangled the different genomic signatures resulting from hybrid origin, backcrossing, and secondary post-origin evolution in the polyploid species. All tetraploids showed a considerable post-origin, species-specific proportion of SNPs. The amount of extant hybridization appears to be related to the degree of geographical and ecological isolation of species. Our data demonstrate that high-quality RAD sequencing data are suitable and highly informative for the analysis of the origin and relationships of polyploid species. The combination of the traditional tools RAxML, STRUCTURE, SplitsTree and recently developed programs like SNAPP, HyDe and SNiPloid established a bioinformatic pipeline for unraveling the complexity of polyploid genomes.
Food authenticity as one of the stimuli to world heritage sites
Authors: Latiff Khairunnisak , Ng Siew Imm , Aziz Yuhanis Abdul , Kamal Basha Norazlyn 
Published: December 2019

PurposeThe purpose of this paper is to discover authenticity dimensions that draw tourists to Melaka and George Town World Heritage Site. It also examined the mediating effect of attachment and the moderating effect of cultural motivation.Design/methodology/approachA mixed-method design was used. Qualitative approach was conducted to discover authenticity dimensions followed by a quantitative approach to explore the dimensions’ ability in predicting attachment and intention to recommend.FindingsFindings revealed that objective, existential and food authenticity were significant stimuli of recommend intention. The mediating effect of attachment on constructive authenticity-intention and existential authenticity-intention were also supported. Cultural motivation indeed moderated the relationship between objective authenticity-attachment, constructive authenticity-attachment and existential authenticity-attachment.Originality/valueKnowing that authenticity can be perceived differently, therefore, a mixed-method study design offers more insight on discovering authenticity elements. Using a qualitative approach, the study began by exploring important authenticity dimensions from both supply and demand groups, and subsequently, these dimensions were verified using quantitative approach. As expected, food authenticity was found as a standalone dimension.
Double-edged effects of ethical leadership in the development of Greater China salespeople's emotional exhaustion and long-term customer relationships
Authors: Lee Chih-Jen , Huang Stanley Y.B. 
Published: December 2019

Purpose
The purpose of this paper is to present how ethical leadership has a double-edged effect to influence emotional exhaustion and long-term customer relationship developments of employees in a sales management context through a latent growth model (LGM).

Design/methodology/approach
To test the LGM, data were collected by surveying 407 salespeople of a retail travel agency with 814 customers in Greater China at multiple points over an eight-month period.

Findings
This study found that, as salespeople perceived more ethical leadership at Time 1, they were more likely to show increases in the work engagement development that increased the service performance development and increased the work–family conflict development over time. In addition, increases in service performance development influenced increases in customer relationship development and increases in work–family conflict development also influenced the emotional exhaustion development.

Originality/value
These findings help managers understand that enabling salespeople to use their full capabilities to their work by ethical leadership may bring not only high service performance but also negative factors that erode salespeople’s well-being.
Modern Public School – technical procurement
Authors: Shaikh Abdul Rehman , Qazi Asad Ali 
Published: December 2019

Learning outcomes
Understand the process of purchasing and procurement in a commercial organization; describe the process of tendering, evaluation of bids, and the selection of the supplier; describe the commercial, technical and financial bids; understand the importance of vendor selection criteria and develop the same; and identify the parameters for the procurement of assets.

Case overview/synopsis
Mr Shaikh, working as Procurement Manager at Modern Public School Sukkur (MPSS), is facing the challenge of procuring the bunk beds for recently inducted students. He was asked by the management of the school and his financiers to procure the best quality bunk beds. These beds shall be used at hostels for the students of Class VI. Director academics had already rejected his initial proposal and requested him to source out some cheaper solution keeping in view the budgets. Mr Shaikh then arranged a sample, which was well within the budget, however, this sample was rejected by the owner of the school on the basis of quality. Mr Shaikh now had to source out not only the best quality product but also a budget-friendly solution. He visited the markets and finalized the best quality of pipes to be used for bunk beds. With limited funds, and very short time, shall he be able to select the vendor of his choice and arrange the best quality products? Shall he be able to motivate his key stakeholders and gain procurement committee approval this time?

Complexity academic level
Bachelor of Business Administration and Master of Business Administration.

Supplementary materials
Teaching Notes are available for educators only. Please contact your library to gain login details or email support@emeraldinsight.com to request teaching notes.

Subject code
CSS 11: Strategy
Thematic analysis of short-term renter preferences for housing market in Kuala Lumpur, Malaysia
Authors: Ho Pui-Yee , Low Sheau-Ting , Wee Siaw-Chui , Choong Weng-Wai 
Published: December 2019

Purpose
The purpose of this paper is to propose a short-term renters’ preference profile for peer-to-peer (P2P) accommodation selection in the housing market in Kuala Lumpur, Malaysia. The emergence of P2P accommodation was initiated by the concept of a sharing economy, allowing the peer provider to share personal properties with the peer end-users. As the housing market has become more competitive, understanding the preference profile among short-term renters is advantageous to offer a matching marketing mix to the targeted customers.

Design/methodology/approach
This study uses qualitative method associated with thematic analysis. A total of 10,509 reviews on Airbnb were gathered from 377 properties in Kuala Lumpur Federal Territory from 2013 to 2017. Thematic analysis assisted by NVivo software was applied to analyse the empirical data. In total, 14 attributes were identified which could be categorised into 5Ps. The concept of 5Ps adopted from modern marketing is commonly used to categorise the strategies in the marketing process. In the current context, 5Ps is used to categorise the preference of the short-term renter in their selection of P2P accommodation including product, price, promotion, process and people.

Findings
This paper has developed a preference profile for P2P accommodation selection in Malaysia’s housing market. In total, 14 attributes were identified and categorised into the 5Ps of marketing mix. The results showed that the majority of short-term renters were concerned mostly with product (73.2 per cent), followed by people (14.5 per cent), process (10.1 per cent), price (2.0 per cent) and promotion (0.2 per cent).

Originality/value
This study contributes to existing literature with a novel case in Malaysia’s housing market by identifying the short-term renter’s preference in the P2P accommodation selection in the Malaysian housing market, specifically in Kuala Lumpur. The preference profile provides guidance for property owners and developers in the housing market to offer the right product in enhancing the marketability and rentability of the property.
“In these streets”: the saliency of place in an alternative black mental health resource centre
Authors: Wainwright John , McKeown Mick , Kinney Malcolm 
Published: December 2019

Purpose
The purpose of this paper is to explore experiences of survivors of the mental health system regularly attending a mental health resource centre predominantly but not exclusively focussed on needs of the BAME community.

Design/methodology/approach
In total, 25 participants took part in a qualitative research study regarding their experiences of mental health and racism, alternative mental health support and struggles in the local black community.

Findings
Issues of race, place and space were central to the experiences of BAME mental health survivors. Participants emphasised the importance of place-based support in their everyday life, with the service provided engendering a sense of belonging conducive to coping with various struggles. Race and racism were also central to these daily struggles and the place of Liverpool 8 was at the core of notions of identity and belonging. The space within the centre provided a sanctuary from the combined discriminations and exclusions attendant on being BAME survivors of the mental health system.

Practical implications
Attention to matters of place and space appears crucial to the articulation of appropriate support.

Social implications
Place is salient to understanding the intersecting identities/experience of racism and mental health discrimination, constituting the basis for a concept of placism; associated with exclusions from feeling safe and included in everyday public places (including within the black community) with the exception of the welcoming and unconditionally accepting space of the centre.

Originality/value
This paper is the first to inquire into place-based experiences of alternative black mental health support. Placism is a novel construct that merits further inquiry and theoretical development.
Emanating the key factors of innovation performance: leveraging on the innovation culture among SMEs in Malaysia
Authors: Hanifah Haniruzila , Abdul Halim Hasliza , Ahmad Noor Hazlina , Vafaei-Zadeh Ali 
Published: December 2019

PurposeInnovation has become an approach to create value for the customer to remain competitive in the market. However, previous research on innovation performance particularly among Bumiputera small and medium-sized enterprises (SMEs) had received little intention. Hence, Bumiputera SMEs need to inculcate the innovation culture to generate innovation performance. As such, the purpose of this study is to examine the ambidextrous orientation and innovation strategy on innovation culture, and how innovation culture could mediate the relationship between ambidextrous orientation and innovation strategy and innovation performance. In addition, this study also examines the role of government support as the moderator between innovation culture and innovation performance.Design/methodology/approachData were collected from 140 Bumiputera SMEs and analyzed using partial least square-structural equation modeling via Smart PLS.FindingsFindings indicated that ambidextrous orientation (alignment and adaptability) and innovation strategy (proactive creativity strategy and growth risk orientation strategy) had a significant impact on innovation culture. Besides, innovation culture mediated the relationship between alignment, proactive creativity strategy, growth risk orientation strategy and innovation performance. Surprisingly, innovation culture does not significantly mediate the relationship between adaptability and innovation performance. However, government support plays an important role to support innovation culture and innovation performance in Bumiputera SMEs.Originality/valueThis study makes both theoretical and practical contributions, especially in identifying the significant role of Bumiputera SMEs in creating an innovation culture. Besides, it explained government support as an important role in strengthening the relationship between innovation culture and innovation performance. The findings of the study will provide great help to Bumiputera entrepreneurs in formulating innovation culture in Malaysian SMEs.
Innovative product marketing strategy: multinational companies in Bangladesh
Authors: Fregidou-Malama Maria , Chowdhury Ehsanul Huda , Hyder Akmal S. 
Published: December 2019

PurposeThis paper aims to deal with international marketing of products, analyzing how adaptation/standardization and network development are achieved when marketing products in Bangladesh.Design/methodology/approachBy applying a qualitative method, the study was conducted at four multinationals, British American Tobacco, Perfetti Van Melle, Tetrapak and Reckitt Benckiser, operating in Bangladesh. Data were collected through semistructured interviews, direct observation and official documents. The analysis was conducted through construction of themes that were identified from the data set.FindingsThe study demonstrates that business relationships related to a local market should be adapted to customer preferences. The research suggests that a balanced combination of product quality and development of new, innovative products adapted to the needs of the market and the customers establishes trust and networks. Cultural and market context were found to influence multinational companies (MNCs)s to standardize the quality of the products and adapt marketing mix components to the needs of consumers.Research limitations/implicationsThe paper contributes to international marketing literature with a model of product marketing based on context, trust, networks and adaptation/standardization. The model introduces the cultural dimension of femininity/ masculinity and the innovation of products and market structure. The study is limited to one emerging market. Further studies should explore other emerging market economies and MNCs.Practical implicationsThe results suggest that to meet the challenges of emerging market economies and achieve success, managers should take people and market needs into consideration.Originality/valueThis paper extends product marketing literature by presenting a context-based model for MNCs’ product marketing.
The Islamic theory of consumer behaviour for ijarah home financing
Authors: Amin Hanudin 
Published: December 2019

Purpose
This study aims to examine the factors that determine consumer behaviour of ijarah home financing using the Islamic theory of consumer behaviour (ITCB). In particular, the specific contract used is al-ijarah muntahia biltamleek. Subsequently, the term al-ijarah muntahia biltamleek home financing is used throughout the paper.

Design/methodology/approach
Drawing upon the ITCB, the present study examines patronage factors for al-ijarah muntahia biltamleek home financing. This study’s data were drawn from valid 409 usable questionnaires.

Findings
Partial least square results suggest that the role of iman, Islamic mortgage support and maqasid consumer index are significantly related to consumer behaviour. In turn, consumer behaviour is instrumental in determining religious satisfaction.

Research limitations/implications
Two limitations were observed that provide direction for future studies. Firstly, the current study is only tested in one particular Islamic home financing product that may explain why the generalisation is an issue. Secondly, the theory is still in its infancy stage, which, of course leads to some criticisms pertinent to its parsimony and applicability to other settings. Despite these limitations, however, this study aims to suggest a so-called Islamic model of factors that can explain consumer behaviour of a particular Islamic home financing product. In particular, scrutinising the product acceptance from the context of the ITCB can open a new debate in this area.

Practical implications
This study indeed reduces the theory–practice gap by suggesting new indicators that are workable to improve consumers’ positive perception of Islamic home financing. Given the results obtained, the ITCB provides a better prediction for consumer behaviour of al-ijarah muntahia biltamleek home financing.

Originality/value
This study expands the applicability of the ITCB to include al-ijarah muntahia biltamleek home financing.