A new plasmid display technology for the in vitro selection of functional phenotype–genotype linked proteins

Background: Display technologies which allow peptides or proteins to be physically associated with the encoding DNA are central to procedures which involve screening of protein libraries in vitro for new or altered function. Here we describe a new system designed specifically for the display of libraries of diverse, functional proteins which utilises the DNA binding protein nuclear factor κB (NF-κB) p50 to establish a phenotype–genotype link between the displayed protein and the encoding gene. Results: A range of model fusion proteins to either the amino- or carboxy-terminus of NF-κB p50 have been constructed and shown to retain the picomolar affinity and DNA specificity of wild-type NF-κB p50. Through use of an optimal combination of binding buffer and DNA target sequence, the half-life of p50–DNA complexes could be increased to over 47 h, enabling the competitive selection of a variety of protein–plasmid complexes with enrichment factors of up to 6000-fold per round. The p50-based plasmid display system was used to enrich a maltose binding protein complex to homogeneity in only three rounds from a binary mixture with a starting ratio of 1:108 and to enrich to near homogeneity a single functional protein from a phenotype–genotype linked Escherichia coli genomic library using in vitro functional selections. Conclusions: A new display technology is described which addresses the challenge of functional protein display. The results demonstrate that plasmid display is sufficiently sensitive to select a functional protein from large libraries and that it therefore represents a useful addition to the repertoire of display technologies.

Optimising employee engagement with internal communication : a social exchange perspective

Drawing on principles of social exchange this thesis employs mediated regression to investigate the relationship between internal communication and employee engagement in the Australian workforce. Findings suggest organisations and supervisors should focus internal communication efforts toward building greater perceptions of support and stronger identification among employees in order to foster optimal engagement. This research contributes to public relations and management scholarship through understanding how perceived support and identification act as mediating mechanisms in the relationship between internal communication and employee engagement at the organisational and supervisory level.

Freedom to choose? Embryo selection, reproductive decision-making and the role of the state

Involving the biopsy of an eight-cell embryo, PGD has been hailed as a means of making reproductive decisions without having to face the heart-wrenching decision to abort an affected foetus. However, controversy around the kinds of traits for which testing can be done, and who has access to the technology, has led to questions about the way in which the technology is developing. Women who are allowed to access in vitro fertilisation (IVF) services can currently also access PGD in limited circumstances.

A new plasmid display technology for the in vitro selection of functional phenotype-genotype linked proteins

Background Display technologies which allow peptides or proteins to be physically associated with the encoding DNA are central to procedures which involve screening of protein libraries in vitro for new or altered function. Here we describe a new system designed specifically for the display of libraries of diverse, functional proteins which utilises the DNA binding protein nuclear factor κB (NF-κB) p50 to establish a phenotype-genotype link between the displayed protein and the encoding gene. Results A range of model fusion proteins to either the amino- or carboxy-terminus of NF-κB p50 have been constructed and shown to retain the picomolar affinity and DNA specificity of wild-type NF-κB p50. Through use of an optimal combination of binding buffer and DNA target sequence, the half-life of p50-DNA complexes could be increased to over 47 h, enabling the competitive selection of a variety of protein-plasmid complexes with enrichment factors of up to 6000-fold per round. The p50-based plasmid display system was used to enrich a maltose binding protein complex to homogeneity in only three rounds from a binary mixture with a starting ratio of 1:108 and to enrich to near homogeneity a single functional protein from a phenotype-genotype linked Escherichia coli genomic library using in vitro functional selections. Conclusions A new display technology is described which addresses the challenge of functional protein display. The results demonstrate that plasmid display is sufficiently sensitive to select a functional protein from large libraries and that it therefore represents a useful addition to the repertoire of display technologies.

Damping structure selection in nonlinear ship manoeuvring models

This paper presents the application of a statistical method for model structure selection of lift-drag and viscous damping components in ship manoeuvring models. The damping model is posed as a family of linear stochastic models, which is postulated based on previous work in the literature. Then a nested test of hypothesis problem is considered. The testing reduces to a recursive comparison of two competing models, for which optimal tests in the Neyman sense exist. The method yields a preferred model structure and its initial parameter estimates. Alternatively, the method can give a reduced set of likely models. Using simulated data we study how the selection method performs when there is both uncorrelated and correlated noise in the measurements. The first case is related to instrumentation noise, whereas the second case is related to spurious wave-induced motion often present during sea trials. We then consider the model structure selection of a modern high-speed trimaran ferry from full scale trial data.

Self-determination as a moderator of demands and control : implications for employee strain and engagement

Does job control act as a stress-buffer when employees' type and level of work self-determination is taken into account? It was anticipated that job control would only be stress-buffering for employees high in self-determined and low in non-self-determined work motivation. In contrast, job control would be stress-exacerbating for employees who were low in self-determined and high in non-self-determined work motivation. Employees of a health insurance organization (N = 123) completed a survey on perceptions of role overload, job control, work self-determination, and a range of strain and engagement indicators. Results revealed that, when individuals high in self-determination perceived high job control, they experienced greater engagement (in the form of dedication to their work). In addition, when individuals high in non-self-determination perceived high job demands, they experienced more health complaints. A significant 3-way interaction demonstrated that, for individuals low in non-self-determination, high job control had the anticipated stress-buffering effect on engagement (in the form of absorption in their work). In addition, low job control was stress-exacerbating. However, contrary to expectations, for those high in non-self-determination, high job control was just as useful as low job control as a stress-buffer. The practical applications of these findings to the organizational context are discussed.

The impact of organisational citizenship behaviour and non-material rewards on dimensions of employee burnout : evidence from the teaching profession

In this chapter, we examine the psychological impact that organisational citizenship behaviours (OCBs) have on individuals performing them. OCB is discretionary employee behaviour that is not systematically rewarded by employers, but that contributes to overall organisational effectiveness (Organ, 1988). In a sample of schoolteachers, we predicted that performing OCBs would differentially impact two dimensions of psychological burnout -personal accomplishment (PA} and emotional exhaustion (EE). Due to the volitional nature of OCB, there are theoretical reasons to suppose that OCB enhances PA. However, it is also possible that certain OCBs constitute increased workload, thereby contributing to a heightened sense of EE. In addition, given prior research showing that non-material rewards such as praise and recognition, lead to positive employee outcomes, we proposed that praise and recognition would strengthen the relationship between OCB and PA, and weaken the relationship between OCB and EE.

An exploratory study into director selection : who do directors want on their boards and how do they select them?

This exploratory study into director selection involved in-depth interviews with Australian non-executive directors to identify what directors consider as important criteria when selecting new members and the approach taken to identify and select candidates. The findings indicate boards select new members based not only on their ability to contribute complementary skills and experience but also on a perceived compatibility with incumbent board members. While these two selection criteria are considered equal in importance, not all selection approaches are able to adequately assess both criteria. As a result many selections fail to realise their selection criteria.

Wheat breeding nurseries, target environments, and indirect selection for grain yield

Use of appropriate nursery environments will maximize gain from selection for yield of wheat (Triticum aestivum L.) in the target population of environments of a breeding program. The objective of this study was to investigate how well-irrigated (low-stress) nursery environments predict yield of lines in target environments that varied in degree of water limitation. Fifteen lines were sampled from the preliminary yield evaluation stage of the Queensland wheat breeding program and tested in 26 trials under on-farm conditions (Target Environments) across nine years (1985 to 1993) and also in 27 trials conducted at three research stations (Nursery Environments) in three years (1987 to 1989). The nursery environments were structured to impose different levels of water and nitrogen (N) limitation, whereas the target environments represented a random sample of on-farm conditions from the target population of environments. Indirect selection and pattern analysis methods were used to investigate selection for yield in the nursery environments and gain from selection in the target environments. Yield under low-stress nursery conditions was an effective predictor of yield under similar low-stress target environments (r = 0.89, P < 0.01). However, the value of the low-stress nursery as a predictor of yield in the water-limited target environments decreased with increasing water stress (moderate stress r = 0.53, P < 0.05, to r = 0.38, P > 0.05; severe stress r = -0.08, P > 0.05). Yield in the stress nurseries was a poor predictor of yield in the target environments. Until there is a clear understanding of the physiological-genetic basis of variation for adaptation of wheat to the water-limited environments in Queensland, yield improvement can best be achieved by selection for a combination of yield potential in an irrigated low-stress nursery and yield in on-farm trials that sample the range of water-limited environments of the target population of environments.

Optimizing employee engagement with internal communication : a social exchange perspective

Employee engagement is linked to higher productivity, lower attrition, and improved organizational reputations resulting in increased focus and resourcing by managers to foster an engaged workforce. While drivers of employee engagement have been identified as perceived support, job characteristics, and value congruence, internal communication is theoretically suggested to be a key influence in both the process and maintenance of employee engagement efforts. However, understanding the mechanisms by which internal communication influences employee engagement has emerged as a key question in the literature. The purpose of this research is to investigate whether social factors, namely perceived support and identification, play a mediating role in the relationship between internal communication and engagement. To test the theoretical model, data are collected from 200 non-executive employees using an online self-administered survey. The study applies linear and mediated regression to the model and finds that organizations and supervisors should focus internal communication efforts toward building greater perceptions of support and stronger identification among employees in order to foster optimal levels of engagement.

Who selects the 'right' directors? An examination of the association between board selection, gender diversity and outcomes

Using a sample of companies from the top 500 listed firms in Australia, we investigate whether the presence of a designated nomination committee and female representation on the nomination committee affect board gender diversity. We also examine whether gender diversity on the board affects firm risk and financial performance. We find that board gender diversity is significantly and positively associated with the presence of a designated nomination committee and that female representation on the nomination committee is a significant explanatory factor of increasing board gender diversity following the release of the 2010 Australian Securities Exchange Corporate Governance Council (ASXCGC) recommendations. Further, our results support the business case for board gender diversity as we find greater gender diversity moderates excessive firm risk which in turn improves firms’ financial performance. Our results are robust after correcting for selection bias and controlling for other board, firm and industry characteristics.

E-mail in the workplace : the role of stress appraisals and normative response pressure in the relationship between email stressors and employee strain

In many organizations, e-mail is an effective and dominant workplace application tool; however, research identifying its role as a potential workplace stressor remains limited. Utilizing the Transactional Model of Stress (Lazarus & Folkman, 1984), 215 full-time administrative and academic staff at a university were surveyed about workplace e-mail. The aim was to study the effects of potential e-mail stressors on emotional exhaustion as mediated and moderated by person and situation variables. Results indicated that 2 distinct e-mail stressors—high quantity and poor quality (in terms of high emotionality and ambiguity) of workplace e-mail—were associated both with stress appraisals (e-mail overload and e-mail uncertainty) and with emotional exhaustion. Furthermore, the effects of the 2 e-mail stressors on emotional exhaustion were mediated by appraised e-mail overload. Perceived normative response pressure—a relevant aspect of the specific work environment—added to the explanation of emotional exhaustion and accentuated the positive effect of e-mail ambiguity on emotional exhaustion, although effects involving normative response pressure were not explained by the stress appraisals.

A review selection method using product feature taxonomy

As of today, online reviews have become more and more important in decision making process. In recent years, the problem of identifying useful reviews for users has attracted significant attentions. For instance, in order to select reviews that focus on a particular feature, researchers proposed a method which extracts all associated words of this feature as the relevant information to evaluate and find appropriate reviews. However, the extraction of associated words is not that accurate due to the noise in free review text, and this affects the overall performance negatively. In this paper, we propose a method to select reviews according to a given feature by using a review model generated based upon a domain ontology called product feature taxonomy. The proposed review model provides relevant information about the hierarchical relationships of the features in the review which captures the review characteristics accurately. Our experiment results based on real world review dataset show that our approach is able to improve the review selection performance according to the given criteria effectively.

Biolistic transformation of elite indica rice (Oryza sativa L.) cultivars through semi-solid and liquid medium selection systems

Following microprojectile mediated delivery of a plasmid construct (pAHC-25) encoding bar (bialophos resistance) gene into five-day-old scutellar calli derived from mature embryos, the effectiveness of selection procedure for bar-gene expressing tissue was compared for two indica rice cultivars (IR-64 and Karnal Local). While IR-64 transformants could be selected through the generally used semi-solid selection medium, the same procedure was not effective in the basmati cultivar Karnal Local. In the latter case, while lower concentrations (2–4 mg 1−1) of the selective agent phosphinothricin (PPT) yielded only escapes, higher concentrations (6–8 mg l−1) inhibited proliferation of transformed as well as untransformed sectors. For Karnal Local, a liquid medium based selection system was successfully utilized for recovering transformed sectors and, eventually, regenerants. The study demonstrates the generation of transformants of two elite indica cultivars using the environment-independent system of mature embryos from seeds.