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.

Progress in epithelial-mesenchymal transition research

The field of research of epithelial-mesenchymal transitions, EMT, and its reverse, mesenchymal-epithelial transitions, MET, has expanded very rapidly indeed from its beginnings, heralded by Professor Betty Hay in the 1970s and 1980s. This expansion has involved the realisation that the EMT was not just an interesting phenomenon of early developmental morphogenetic cell behaviour, but bore remarkable resemblance to clinically crucial pathological events in cancer invasion. Not surprisingly, this discipline soon became numerically dominant in the EMT publication field. Simultaneously, the EMT concept has been extended to normal physiological wound healing. Exploration revealed that these resemblances were more than skin deep: the same sets of growth factors, receptors, transcription factors, epigenetic marks and signalling pathways turned up repeatedly in EMTs and METs in a variety of contexts, both pathological and normal. This molecular genetic research in turn uncovered similarities of the EMT signature to that of fibrosis, a set of diseases which is of enormous clinical importance, rivalling that of cancer. Most recently, and more surprisingly, the EMT signature has shown considerable similarity to that found in stem cell and cancer stem cell biology.

Globalising the body : globalisation and reproductive rights

Globalisation is a phenomenon of the contemporary world. Everywhere around us there seem to be signs of the power of the forces of globalisation: in our media and popular culture; in our international linkages across continents through international travel and telecommunications; in our globalised trade; and with the global movement of people, a process which itself ranges from the movement of international tourists to the international movement of refugees and other displaced persons. The processes of globalisation seem to simultaneously unify and divide us. There is no doubt that we live in a globalised world and that we are connected to others in previously unimaginable ways by transportation, telecommunications and economics. Yet, while this global context increasingly links us to others, there is also a very real sense in which separation, difference and the local have also gained a new significance; we are locked in a tension between the universal and the particular that has come to typify contemporary society. This article explores the meanings of globalisation and this dynamic – or tension – between the universal and the particular in terms of its implications for the body and, in particular, its significance for women and their reproductive rights.

Apple skin patterning is associated with differential expression of MYb10

Background Some apple (Malus × domestica Borkh.) varieties have attractive striping patterns, a quality attribute that is important for determining apple fruit market acceptance. Most apple cultivars (e.g. 'Royal Gala') produce fruit with a defined fruit pigment pattern, but in the case of 'Honeycrisp' apple, trees can produce fruits of two different kinds: striped and blushed. The causes of this phenomenon are unknown. Results Here we show that striped areas of 'Honeycrisp' and 'Royal Gala' are due to sectorial increases in anthocyanin concentration. Transcript levels of the major biosynthetic genes and MYB10, a transcription factor that upregulates apple anthocyanin production, correlated with increased anthocyanin concentration in stripes. However, nucleotide changes in the promoter and coding sequence of MYB10 do not correlate with skin pattern in 'Honeycrisp' and other cultivars differing in peel pigmentation patterns. A survey of methylation levels throughout the coding region of MYB10 and a 2.5 Kb region 5' of the ATG translation start site indicated that an area 900 bp long, starting 1400 bp upstream of the translation start site, is highly methylated. Cytosine methylation was present in all three contexts, with higher methylation levels observed for CHH and CHG (where H is A, C or T) than for CG. Comparisons of methylation levels of the MYB10 promoter in 'Honeycrisp' red and green stripes indicated that they correlate with peel phenotypes, with an enrichment of methylation observed in green stripes. Conclusions Differences in anthocyanin levels between red and green stripes can be explained by differential transcript accumulation of MYB10. Different levels of MYB10 transcript in red versus green stripes are inversely associated with methylation levels in the promoter region. Although observed methylation differences are modest, trends are consistent across years and differences are statistically significant. Methylation may be associated with the presence of a TRIM retrotransposon within the promoter region, but the presence of the TRIM element alone cannot explain the phenotypic variability observed in 'Honeycrisp'. We suggest that methylation in the MYB10 promoter is more variable in 'Honeycrisp' than in 'Royal Gala', leading to more variable color patterns in the peel of this cultivar.

Personal media and wireless cities : towards an urban spatial analysis

For the past decade, at least, varieties of small, hand held networked instruments have appeared on the global scene, selling in record numbers, and being utilized by all manner of persons from the old to the young; children, women, men, the wealthy and the poor and in all countries. Their presences bespeak a radical shift in telecommunications infrastructure and the future of communications. They are particularly visible in urban areas where mobile transmission network infrastructure (3G, 4G, cellular and Wi-Fi) is more established and substantial, options more plentiful, and density of populations more dramatic. These end user products—I phones, cell phones, Blackberries, DSi, DS, IPads, Zooms, and others – of the mobile communications industry are the latest, hottest globalized commodities. At the same time, wirelessness, or the state of being wireless, and therefore capable of taking along one's networks, communicating from unlikely spaces, and navigating with GPS, is a complex social, political and economic communications phenomenon of early 21st century life. This thesis examines the specter of being wireless in cities. It lends the entire idea an experimentally envisioned, historical and planned context wherein personalization of media tools is seen both as a design development of corporate, artistic, and military imagination, as well as a profound social phenomenon enabling new forms of sharing, belonging, and urban community. In doing that it asserts the parameters of a new mobile space which, aside from clear benefits to humankind by way of mobility, has reinscribed numerous categories including gender. Moreover, it posits the recognition of other, more nuanced theoretical spaces for complex readings of gender and gendered use, including some instantiation of the notion of 'network' itself as a cyborgian and gendered social form. Additionally, cities are studied as places where technology is not only quickly popularized, but is connected to larger political interests, such as the reading of data, tracking of information, and the new security culture. In so doing the work has been undertaken as an urban spatial analysis and experimental ethnography, utilizing architectural, feminist, techno-utopian, industrial and theoretical literatures as discursive underpinnings from whence understandings and interpretations of mobile space, the mobile office, networked mobility, and personal media have come, linking the space of cities to specific, pioneering urban public art projects in which voice, texting and MMS have been utilized in expressions of ubiquitous networks and urban history. Through numerous examples of techno art, the thesis discusses the 'wireless city' as an emerging cultural, socially constructed economic and spatial entity, both conceived and formed through historic processes of urbanization.

Molecular sliding filament model for muscular contraction based on multiscale investigation

A multiscale approach that bridges the biophysics of the actin molecules at nanoscale and the biomechanics of actin filament at microscale level is developed and used to evaluate the mechanical performances of actin filament bundles. In order to investigate the contractile properties of skeletal muscle which is induced by the protein motor of myosin, a molecular model is proposed in the prediction of the dynamic behaviors of skeletal muscle based on classic sliding filament model. Randomly distributed myosin motors are applied on a 2.2 μm long sarcomere, whose principal components include actin and myosin filaments. It can be found that, the more myosin motors on the sarcomere, the faster the sarcomere contracts. The result demonstrates that the sarcomere shortening speed cannot increase infinitely by the modulation of myosin, thus providing insight into the self-protective properties of skeletal muscles. This molecular filament sliding model provides a theoretical way to evaluate the properties of skeletal muscles, and contributes to the understandings of the molecular mechanisms in the physiological phenomenon of muscular contraction.

Civil litigation limitation periods : submission to the Royal Commission into institutional responses to child sexual abuse

This submission makes one simple yet powerful recommendation for law reform to promote justice for survivors of child sexual abuse. It is informed by extensive analyses of the phenomenon of child sexual abuse and its psychological sequelae, legislative time limits and case law across Australia and internationally, the policy reasons underpinning statutory time limits generally, and the need for fairness, certainty and practicability in the legal system. The recommendation is that legislative reform is required in all Australian States and Territories to remove time limitations for civil claims for injuries caused by child sexual abuse.

Stabilization of parametric roll resonance with active u-tanks via Lyapunov control design

Parametric ship roll resonance is a phenomenon where a ship can rapidly develop high roll motion while sailing in longitudinal waves. This effect can be described mathematically by periodic changes of the parameters of the equations of motion, which lead to a bifurcation. In this paper, the control design of an active u-tank stabilizer is carried out using Lyapunov theory. A nonlinear backstepping controller is developed to provide global exponential stability of roll. An extension of commonly used u-tank models is presented to account for large roll angles, and the control design is tested via simulation on a high-fidelity model of a vessel under parametric roll resonance.

Transmedia Practice : A Collective Approach

This volume introduces a collective approach that positions transmedia as a dynamic phenomenon which undergoes constant innovation as it absorbs current trends and advances in its constituent disciplines. The first section, 'Sustaining Future Practices', explores emerging models for defining stakeholder needs, understanding resource requirements and measuring the value and success of transmedia productions. The focus then shifts to 'Intersecting Contexts of Transmedia Practices', which uses the juxtaposition of a diverse collection of case studies to transcend not only the debates about how to define transmedia, but also the professional and disciplinary boundaries that impose artificial constraints upon the way transmedia projects are approached and understood. This inter-disciplinary dialogue aims to promote an ongoing conversation on the challenges and opportunities associated with sustaining this vital creative industry.

Developing a framework of effective prevention and response strategies in workplace sexual harassment

Sexual harassment remains a widespread workplace phenomenon, despite laws that proscribe it. Drawing initially on a typology from the violence prevention literature that conceptualizes prevention and response approaches according to when they occur, the paper synthesizes strategies identified in literature addressing workplace sexual harassment, as well as other workplace injustices or grievances. The paper utilizes this previous research to develop a framework of sexual harassment prevention strategies along two dimensions: functions and timing. The framework offers a research-informed set of organization-wide preventative and remedial approaches, a systemic approach to what is often seen as an individual problem, and a means to better focus interventions that are often disparate and unco-ordinated. The paper also highlights important areas for future research including a stronger focus on longer-term (tertiary) corrective actions.

The inter-relationships between ovarian-independent growth, tumorigenicity, invasiveness and antioestrogen resistance in the malignant progression of human breast cancer

Among the processes contributing to the progressive acquisition of the highly malignant phenotype in breast cancer are ovarian-independent growth, antioestrogen resistance and increased metastatic potential. We have previously observed that increased invasiveness and development of ovarian-independent growth occur independently. In an attempt to define the inter-relationships between these processes further, we have compared the phenotypes of ovarian-independent, invasive and antioestrogen-resistant sublines of the ovarian-dependent human breast cancer cell line MCF-7. Cells acquiring ovarian-independent growth can retain sensitivity to anti-oestrogens. One clone of MCF-7 cells selected for stable antioestrogen resistance has become non-tumorigenic but its invasive potential remains unaltered. Thus, acquisitions of some characteristics of the progressed phenotype can occur independently. This phenomenon of independent parameters in phenotypic progression could partly explain the considerable intra- and intertumour heterogeneity characteristic of breast tumours.

Engagement as dialogue : presenting a pragmatic perspective

‘Engagement’ is one of the buzzwords of 21st century public relations theory and practice. Yet the concept of engagement remains frustratingly nebulous and intangible, lacking clear definition and consistent use. This paper suggests that the concept of dialogue can provide public relations practitioners and academics with a framework for defining engagement that allows deep insights into the range of ideas and contexts it covers. Specifically, this paper argues for the use of a pragmatic practitioner perspective on dialogue as a lens through which to study engagement. Current literature clearly articulates the normative and prescriptive perspectives on dialogue in public relations, but leaves the story of the reality of the practitioner experience of dialogue largely untold. New research tells this ‘missing’ story of the practitioner perspective on dialogue, which in turn offers some insights into the forms and functions of engagement in practice. Dialogue is seen as encompassing three different types of public relations practice: informing stakeholders of organisational decisions and receiving their feedback; consulting with stakeholders on the strategies and tactics used by organisations to achieve their chosen goals; and including stakeholder input in the making of decisions on what organisational goals should be. Adopting this pragmatic practitioner perspective on dialogue to view engagement provides scaffolding that is strong enough to encourage development of a consistent definition of its meaning; while still allowing the freedom and scope necessary to develop deep and rich understanding of the phenomenon.

Developing a multidimensional scale of customer-oriented deviance (COD)

Although frontline employees' bending of organizational rules and norms for customers is an important phenomenon, marketing scholars to date only broadly describe over-servicing behaviors and provide little distinction among deviant behavioral concepts. Drawing on research on pro-social and pro-customer behaviors and on studies of positive deviance, this paper develops and validates a multi-faceted, multi-dimensional construct term customer-oriented deviance. Results from two samples totaling 616 frontline employees (FLEs) in the retail and hospitality industries demonstrate that customer-oriented deviance is a four-dimensional construct with sound psychometric properties. Evidence from a test of a theoretical model of key antecedents establishes nomological validity with empathy/perspective-taking, risk-taking propensity, role conflict, and job autonomy as key predictors. Results show that the dimensions of customer-oriented deviance are distinct and have significant implications for theory and practice.

Business planning in emerging firms : uses and effects

Business planning is at the core of entrepreneurship as it has implications for opportunity discovery and exploitation. This thesis' objectives are to disentangle the relationships between business planning and venture emergence to reconcile previous inconsistent findings. It reveals that the formalization of planning, the effort invested in the venture and the revision of the plan influence success for entrepreneurs in the process of launching their firm. This thesis provides generalizable results about the phenomenon of business planning by using a longitudinal random sample of emerging firms.

Detection of object onset and offset in naturalistic scenes

The present study was conducted to investigate whether ob- servers are equally prone to overlook any kinds of visual events in change blindness. Capitalizing on the finding from visual search studies that abrupt appearance of an object effectively captures observers' attention, the onset of a new object and the offset of an existing object were contrasted regarding their detectability when they occurred in a naturalistic scene. In an experiment, participants viewed a series of photograph pairs in which layouts of seven or eight objects were depicted. One object either appeared in or disappeared from the layout, and participants tried to detect this change. Results showed that onsets were detected more quickly than offsets, while they were detected with equivalent ac- curacy. This suggests that the primacy of onset over offset is a robust phenomenon that likely makes onsets more resistant to change blindness under natural viewing conditions.