Non-combinatorial library screening reveals subsite cooperativity and identifies new high-efficiency substrates for kallikrein-related peptidase 14

An array of substrates link the tryptic serine protease, kallikrein-related peptidase 14 (KLK14), to physiological functions including desquamation and activation of signaling molecules associated with inflammation and cancer. Recognition of protease cleavage sequences is driven by complementarity between exposed substrate motifs and the physicochemical signature of an enzyme's active site cleft. However, conventional substrate screening methods have generated conflicting subsite profiles for KLK14. This study utilizes a recently developed screening technique, the sparse matrix library, to identify five novel high-efficiency sequences for KLK14. The optimal sequence, YASR, was cleaved with higher efficiency (k(cat)/K(m)=3.81 ± 0.4 × 10(6) M(-1) s(-1)) than favored substrates from positional scanning and phage display by 2- and 10-fold, respectively. Binding site cooperativity was prominent among preferred sequences, which enabled optimal interaction at all subsites as indicated by predictive modeling of KLK14/substrate complexes. These simulations constitute the first molecular dynamics analysis of KLK14 and offer a structural rationale for the divergent subsite preferences evident between KLK14 and closely related KLKs, KLK4 and KLK5. Collectively, these findings highlight the importance of binding site cooperativity in protease substrate recognition, which has implications for discovery of optimal substrates and engineering highly effective protease inhibitors.

Comparison of key competences of DB clients and contractors in the construction market of the People’s Republic of China

Clients and Design-build (DB) contractors are two key stakeholders in DB projects, and contribute significantly to the successful project performance. This study aims to identify and compare such key competences in the construction market of the PRC. After the survey of available literature and face-to-face interviews, a two-round Delphi questionnaire survey was conducted to identify the key competences of clients and DB contractors in DB projects. Relative importance of these identified competences were ranked and compared. The questionnaire results indicated distinct differences between the key competences of clients and that of contractors. The contractor’s key competences emphasize on DB experience, corporate management capability, building and design expertise, financial capability, enterprise qualification and reputation. While the client’s competences focus on the ability to clearly define the project scope & requirements, financial capacity, contract management ability, adequate staff, effective coordination with DB contractor and similar DB experience. Both clients and DB contractors should clearly understand the competence requirements in DB projects and possess all the necessary competences for the successful outcome of DB projects. The identification of these key competences provides clients and DB contractors with indicators to assess their capabilities before going for the DB option. Furthermore, the comparison of competences for clients and DB contractors will result in better understanding of DB system and improve the communication between these stakeholders.

Critical Factors of Promoting Market Demand of Sustainable Housing in Australia

Strong regulatory pressure on environmental issues and the improved public awareness will continue to influence the market demand for sustainable housing in the coming years. Despite this potential, the voluntary up-take rate of sustainable practices is not as high as expected within the new built housing industry. This is in contrast to the influx of emerging building technologies, new materials and innovative designs as seen in office buildings and exemplar homes built worldwide. One possible reason for this is that key stakeholders such as developers, builders and consumers do not fully understand and appreciate the tangible and mutual benefits of sustainability in their professional and business activities. This situation warrants the study of a multifaceted strategy that integrates the needs of multiple stakeholders. This research investigates multiple factors that affect key stakeholder’s benefits in sustainable housing implementation. Drawing insights from a quantitative study on a questionnaire survey and a qualitative study of in-depth interviews with key stakeholders in the Australian housing industry, 11 critical factors of driving market demand for sustainable housing were unearthed. Their inter-relationships were identified with the aid of Interpretive Structural Modelling. The study concludes with a hierarchical model that amalgamates the strategies for the decision making of key stakeholders.

Population structure of Bactrocera dorsalis s.s., B. papayae and B. philippinensis (Diptera: Tephritidae) in southeast Asia: evidence for a single species hypothesis using mitochondrial DNA and wingshape data

Background Bactrocera dorsalis s.s. is a pestiferous tephritid fruit fly distributed from Pakistan to the Pacific, with the Thai/Malay peninsula its southern limit. Sister pest taxa, B. papayae and B. philippinensis, occur in the southeast Asian archipelago and the Philippines, respectively. The relationship among these species is unclear due to their high molecular and morphological similarity. This study analysed population structure of these three species within a southeast Asian biogeographical context to assess potential dispersal patterns and the validity of their current taxonomic status. Results Geometric morphometric results generated from 15 landmarks for wings of 169 flies revealed significant differences in wing shape between almost all sites following canonical variate analysis. For the combined data set there was a greater isolation-by-distance (IBD) effect under a ‘non-Euclidean’ scenario which used geographical distances within a biogeographical ‘Sundaland context’ (r2 = 0.772, P < 0.0001) as compared to a ‘Euclidean’ scenario for which direct geographic distances between sample sites was used (r2 = 0.217, P < 0.01). COI sequence data were obtained for 156 individuals and yielded 83 unique haplotypes with no correlation to current taxonomic designations via a minimum spanning network. BEAST analysis provided a root age and location of 540kya in northern Thailand, with migration of B. dorsalis s.l. into Malaysia 470kya and Sumatra 270kya. Two migration events into the Philippines are inferred. Sequence data revealed a weak but significant IBD effect under the ‘non-Euclidean’ scenario (r2 = 0.110, P < 0.05), with no historical migration evident between Taiwan and the Philippines. Results are consistent with those expected at the intra-specific level. Conclusions Bactrocera dorsalis s.s., B. papayae and B. philippinensis likely represent one species structured around the South China Sea, having migrated from northern Thailand into the southeast Asian archipelago and across into the Philippines. No migration is apparent between the Philippines and Taiwan. This information has implications for quarantine, trade and pest management.

Market orientation in the nonprofit sector

As the nonprofit sector moves into a more competitive environment it is being required by the community to become more efficient and effective. One response is for nonprofit organisations to become market oriented, which is the familiar response in the for-profit sector. Two components of market orientation, that is market segmentation and customer oriented products, fit well within the peculiarities of a nonprofit organisation. This is usually accompanied by the desire to obtain a competitive advantage causes problems for various stakeholders within the organisation. This paper contends that three factors, management, scarcity of resources, and conflict between organisational objectives and market orientation, are major influences on the adoption of a market oriented culture for a nonprofit organisation.

Using cases in a mixed learning environment : meeting the challenges of higher education reform

This paper explains the context for pedagogy in a specific undergraduate course; the comparative benefits of using cases in a mixed learning environment (simultaneous large and small groups); and illustrates one significant way for universities to respond to increasing demand for delivery efficiency while maintaining high quality learning outcomes. Thus, to achieve objectives of the subject, tutorial classes expand on what is taught in lectures and provide the necessary context to analyse cases in more detail. A small, qualitative study explored experiences of market research tutors with the use of case method teaching reported. Implications of the study for case teaching in higher education are identified.

Disability and the labour market : an analysis of British males

This paper attempts, using data from the British Labour Force Survey 1996, to examine to what extent differences in labour market outcomes between able-bodied and disabled men may be attributed to differences in endowments of human capital and associated productivity differences. Both labour force participation and selectivity corrected human capital equations are estimated and decomposition techniques applied to them. Using the methodology of Baldwin and Johnson [Baldwin, M., Johnson, W.G., 1994. Labor market discrimination against men with disabilities. Journal of Human Resources, XXIX(1), Winter, 1–19], the employment effects of wage discrimination against the disabled are also estimated. Evidence of both substantial wage and participation rate differences between able-bodied and disabled men are found, which have implications for the operation of the 1995 Disability Discrimination Act.