Mechanism‐based selection of a potent kallikrein‐related peptidase 7 inhibitor from a versatile library based on the sunflower trypsin inhibitor SFTI‐1

Potent and specific enzyme inhibition is a key goal in the development of therapeutic inhibitors targeting proteolytic activity. The backbone-cyclized peptide, Sunflower Trypsin Inhibitor (SFTI-1) affords a scaffold that can be engineered to achieve both these aims. SFTI-1's mechanism of inhibition is unusual in that it shows fast-on/slow-off kinetics driven by cleavage and religation of a scissile bond. This phenomenon was used to select a nanomolar inhibitor of kallikrein-related peptidase 7 (KLK7) from a versatile library of SFTI variants with diversity tailored to exploit distinctive surfaces present in the active site of serine proteases. Inhibitor selection was achieved through the use of size exclusion chromatography to separate protease/inhibitor complexes from unbound inhibitors followed by inhibitor identification according to molecular mass ascertained by mass spectrometry. This approach identified a single dominant inhibitor species with molecular weight of 1562.4 Da, which is consistent with the SFTI variant SFTI-WCTF. Once synthesized individually this inhibitor showed an IC50 of 173.9 ± 7.6 nM against chromogenic substrates and could block protein proteolysis. Molecular modeling analysis suggested that selection of SFTI-WCTF was driven by specific aromatic interactions and stabilized by an enhanced internal hydrogen bonding network. This approach provides a robust and rapid route to inhibitor selection and design.

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.

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.

Indexes of insulin resistance and secretion in obese children and adolescents : a validation study

OBJECTIVE To assess the concurrent validity of fasting indexes of insulin sensitivity and secretion in - obese prepubertal (Tanner stage 1) children and pubertal (Tanner stages 2-5) glucose tolerance test (FSIVGTT) as a criterion measure. RESEARCH DESIGN AND METHODS Eighteen obese children and adolescents (11 girls and 7 boys, mean age 12.2 +/- 2.4 years, mean BMI 35.4 +/- 6.2 kg/m(2), mean BMI-SDS 3.5 +/- 0.5, 7 prepubertal and I I pubertal) participated in the study. All participants underwent an insulin-modified FSIVGTT on two occasions, and 15 repeated this test a third time (mean 12.9 and 12.0 weeks apart). S-i measured by the FSIVGTT was compared with homeostasis model assessment (HOMA) of insulin resistance (HOMA-IR), quantitative insulin-sensitivity check index (QUICKI), fasting glucose-to-insulin ratio (FGIR), and fasting insulin (estimates of insulin sensitivity derived from fasting samples). The acute insulin response (AIR) measured by the FSIVGTT was compared with HOMA of percent beta-cell function (HOMA-beta%), FGIR, and fasting insulin (estimates of insulin secretion derived from fasting samples). RESULTS There was a significant negative correlation between HOMA-IR and S-i (r = -0.89, r = -0.90, and r = -0.81, P < 0.01) and a significant positive correlation between QUICKI and S-i (r = 0.89, r = 0.90, and r = 0.81, P < 0.01) at each time point. There was a significant positive correlation between FGIR and S-i (r = 0.91, r = 0.91, and r = 0.82, P < 0.01) and a significant negative correlation between fasting insulin and S-i (r = -90, r = -0.90, and r = -0.88, P < 0.01). HOMA-beta% was not as strongly correlated with AIR (r = 0.60, r = 0.54, and r = 0.61, P < 0.05). CONCLUSIONS HOMA-IR, QUICKI, FGIR, and fasting insulin correlate strongly with S-i assessed by the FSIVGTT in obese children and adolescents. Correlations between HOMA-β% FGIR and fasting insulin, and AIR were not as strong. Indexes derived from fasting samples are a valid tool for assessing insulin sensitivity in prepubertal and pubertal obese children.

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.

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.

Review: "Measuring and evaluating sustainability : ethics in sustainability indexes" by Sarah E. Fredericks, New York, Routledge, Routledge Studies in Sustainable Development, 2014

The concept of ‘sustainability’ has been pushed to the forefront of policy-making and politics as the world wakes up to the impacts of climate change and the effects of the rapid urbanisation and modern urban lifestyles (Yigitcanlar and Teriman 2014). Climate change and fossil fuel-based energy policy have emerged as the biggest challenges for our planet, threatening both built and natural systems with long-term consequences. However, the threats are not limited to the impacts of climate change and unsustainable energy system only – e.g., impacts of rapid urbanisation, socioeconomic crises and governance hiccups are just to name a few (Yigitcanlar 2010a). Along with these challenges, successfully coping with the enormous transformations that our cities, societies and the environment have been going through during the last few decades, and their...

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.

Do socially responsible investment indexes outperform conventional indexes?

The question of whether more Socially Responsible (SR) firms outperform or underperform other conventional firms has been debated in the economic literature. In this study, using the Socially Responsible Investment (SRI) indexes and conventional stock indexes in the US, the UK and Japan, first and second moments of firm performance distributions are estimated based on the Markov Switching (MS) model. We find two distinct regimes (bear and bull) in the SRI markets as well as the stock markets for all the three countries. These regimes occur with the same timing in both types of market. No statistical difference in means and volatilities generated from the SRI indexes and conventional indexes in either region was found. Furthermore, we find strong comovements between the two indexes in both the regimes.

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.

Reflections on issues of power in packaged software selection

The adoption of packaged software is becoming increasingly common in a variety of organizations and much of the packaged software literature presents this as a straightforward, linear process based on rationalistic evaluation. This paper applies the framework of power relations developed by Markus and Bjørn-­‐Anderson (1987) to a longitudinal study concerning the adoption of a customer relationship management package in a small organization. This is used to highlight both overt and covert power issues within the selection and procurement of the product and illustrate the interplay of power between senior management, IT managers, IT vendors and consultants, and end-­‐users. The paper contributes to the growing body of literature on packaged software and also to our understanding of how power is deeply embedded within the surrounding processes.

IT consultants, salesmanship and the challenges of packaged software selection in SMEs

Purpose – This paper seeks to analyse the process of packaged software selection in a small organization, focussing particularly on the role of IT consultants as intermediaries in the process. Design/methodology/approach – This is based upon a longitudinal, qualitative field study concerning the adoption of a customer relationship management package in an SME management consultancy. Findings – The authors illustrate how the process of “salesmanship”, an activity directed by the vendor/consultant and focussed on the interests of senior management, marginalises user needs and ultimately secures the procurement of the software package. Research limitations/implications – Despite the best intentions the authors lose something of the rich detail of the lived experience of technology in presenting the case study as a linear narrative. Specifically, the authors have been unable to do justice to the complexity of the multifarious ways in which individual perceptions of the project were influenced and shaped by the opinions of others. Practical implications – Practitioners, particularly those from within SMEs, should be made aware of the ways in which external parties may have a vested interest in steering projects in a particular direction, which may not necessarily align with their own interests. Originality/value – This study highlights in detail the role of consultants and vendors in software selection processes, an area which has received minimal attention to date. Prior work in this area emphasises the necessary conditions for, and positive outcomes of, appointing external parties in an SME context, with only limited attention being paid to the potential problems such engagements may bring.