Mastering the canonical loop of serine protease inhibitors : enhancing potency by optimising the internal hydrogen bond network

Background Canonical serine protease inhibitors commonly bind to their targets through a rigid loop stabilised by an internal hydrogen bond network and disulfide bond(s). The smallest of these is sunflower trypsin inhibitor (SFTI-1), a potent and broad-range protease inhibitor. Recently, we re-engineered the contact β-sheet of SFTI-1 to produce a selective inhibitor of kallikrein-related peptidase 4 (KLK4), a protease associated with prostate cancer progression. However, modifications in the binding loop to achieve specificity may compromise structural rigidity and prevent re-engineered inhibitors from reaching optimal binding affinity. Methodology/Principal Findings In this study, the effect of amino acid substitutions on the internal hydrogen bonding network of SFTI were investigated using an in silico screen of inhibitor variants in complex with KLK4 or trypsin. Substitutions favouring internal hydrogen bond formation directly correlated with increased potency of inhibition in vitro. This produced a second generation inhibitor (SFTI-FCQR Asn14) which displayed both a 125-fold increased capacity to inhibit KLK4 (Ki = 0.0386±0.0060 nM) and enhanced selectivity over off-target serine proteases. Further, SFTI-FCQR Asn14 was stable in cell culture and bioavailable in mice when administered by intraperitoneal perfusion. Conclusion/Significance These findings highlight the importance of conserving structural rigidity of the binding loop in addition to optimising protease/inhibitor contacts when re-engineering canonical serine protease inhibitors.

Tests of LiteSteel Beams subject to combined shear and bending actions

This paper presents the details of an experimental study of a cold-formed steel beam known as LiteSteel Beam (LSB) subject to combined shear and bending actions. The LSBs have the beneficial characteristics of torsionally rigid rectangular hollow flanges combined with economical fabrication processes from a single strip of high strength steel. They combine the stability of hot-rolled steel sections with the high strength to weight ratio of conventional cold-formed steel sections. The LSB sections are commonly used as flexural members in residential, industrial and commercial buildings. In order to ensure safe and efficient designs of LSBs, many research studies have been undertaken on the flexural and shear strengths of LSBs. To date, however, no investigation has been conducted into the strength of LSB sections under combined shear and bending actions. Hence a detailed experimental study involving 18 tests was undertaken to investigate the behaviour and strength of LSBs under combined shear and bending actions. Test results showed that AS/NZS 4600 design rules for unstiffened webs grossly underestimated the capacity of LSBs. Therefore improved design equations were proposed for the combined shear and bending capacities of LSBs based on experimental results.

The effect of intraoperative force on scoliosis correction at the apex : a biomechanical investigation

Adolescent idiopathic scoliosis (AIS) is a complex 3D deformity of the spine, which may require surgical correction in severe cases. Computer models of the spine provide a potentially powerful tool to virtually ‘test’ various surgical scenarios prior to surgery. Using patient-specific computer models of seven AIS patients who had undergone a single rod anterior procedure, we have recently found that the majority of the deformity correction occurs at the apical joint or the joint immediately cephalic to the apex. In the current paper, we investigate the biomechanics of the apical joint for these patients using clinically measured intra-operative compressive forces applied during implant placement. The aim of this study is to determine a relationship between the compressive joint force applied intra-operatively and the achievable deformity correction at the apical joint.

A computer model to simulate scoliosis surgery

Use of patient-specific computer models as a pre-operative planning tool permits predictions of the likely deformity correction and allows a more detailed investigation of the biomechanical influence of different surgical procedures on the scoliotic spinal anatomy. In this paper, patient-specific computer models are used of adolescent idiopathic scoliosis patients who underwent a single rod anterior procedure at the Mater Children’s Hospital in Brisbane, to predict deformity correction and to investigate the change in biomechanics of the scoliotic spine due to surgical compressive forces applied during implant placement.

Sentencing and public confidence : results from a national Australian survey on public opinions towards sentencing

This paper examines the critical issue of public confidence in sentencing, and presents findings from Phase I of an Australia-wide sentencing and public confidence project. Phase I comprised a nationally representative telephone survey of 6005 participants. The majority of respondents expressed high levels of punitiveness and were dissatisfied with sentences imposed by the courts. Despite this, many were strongly supportive of the use of alternatives to imprisonment for a range of offences. These nuanced views raise questions regarding the efficacy of gauging public opinion using opinion poll style questions; indeed the expected outcome from this first phase of the four phase sentencing and public confidence project. The following phases of this project, reported on elsewhere, examined the effects of various interventions on the robustness and nature of these views initially expressed in a standard ‘top of the head’ opinion poll.

Australian journalism studies after ‘Journalism’: breaking down the disciplinary boundaries (for good)

This paper argues that if journalism is to remain a relevant and dynamic academic discipline, it must urgently reconsider the constrained, heavily-policed boundaries traditionally placed around it (particularly in Australia). A simple way of achieving this is to redefine its primary object of study: away from specific, rigid, professional inputs, towards an ever-growing range of media outputs. Such a shift may allow the discipline to freely re-assess its pedagogical and epistemological relationships to contemporary newsmaking practices (or, the ‘new’ news).

Legal issues related to Accountable-eHealth systems in Australia

Information privacy requirements of patients and information requirements of healthcare providers (HCP) are competing concerns. Reaching a balance between these requirements have proven difficult but is crucial for the success of eHealth systems. The traditional approaches to information management have been preventive measures which either allow or deny access to information. We believe that this approach is inappropriate for a domain such as healthcare. We contend that introducing information accountability (IA) to eHealth systems can reach the aforementioned balance without the need for rigid information control. IA is a fairly new concept to computer science, hence; there are no unambiguously accepted principles as yet. But the concept delivers promising advantages to information management in a robust manner. Accountable-eHealth (AeH) systems are eHealth systems which use IA principles as the measure for privacy and information management. AeH systems face three main impediments; technological, social and ethical and legal. In this paper, we present the AeH model and focus on the legal aspects of AeH systems in Australia. We investigate current legislation available in Australia regarding health information management and identify future legal requirements if AeH systems are to be implemented in Australia.

Krüppel-associated box (KRAB)-associated co-repressor (KAP-1) Ser-473 phosphorylation regulates heterochromatin protein 1 (HP1- ) mobilization and DNA repair in heterochromatin

The DNA damage response encompasses a complex series of signaling pathways that function to regulate and facilitate the repair of damaged DNA. Recent studies have shown that the repair of transcriptionally inactive chromatin, named heterochromatin, is dependent upon the phosphorylation of the co-repressor, Krüppel-associated box (KRAB) domain-associated protein (KAP-1), by the ataxia telangiectasia-mutated (ATM) kinase. Co-repressors, such as KAP-1, function to regulate the rigid structure of heterochromatin by recruiting histone-modifying enzymes, such HDAC1/2, SETDB1, and nucleosome-remodeling complexes such as CHD3. Here, we have characterized a phosphorylation site in the HP1-binding domain of KAP-1, Ser-473, which is phosphorylated by the cell cycle checkpoint kinase Chk2. Expression of a nonphosphorylatable S473A mutant conferred cellular sensitivity to DNA-damaging agents and led to defective repair of DNA double-strand breaks in heterochromatin. In addition, cells expressing S473A also displayed defective mobilization of the HP1-β chromodomain protein. The DNA repair defect observed in cells expressing S473A was alleviated by depletion of HP1-β, suggesting that phosphorylation of KAP-1 on Ser-473 promotes the mobilization of HP1-β from heterochromatin and subsequent DNA repair. These results suggest a novel mechanism of KAP-1-mediated chromatin restructuring via Chk2-regulated HP1-β exchange from heterochromatin, promoting DNA repair.

Dichotomy in the design studio : adapting to new blended learning environments

Since the architectural design studio learning environment was first established in the early 19th century at the École des Beaux-Arts in Paris, there has been a complete transformation in how the discipline of architecture is practiced and how students of architecture acquire information. Digital technologies allow students to access information instantly and learning is no longer confined to the rigid boundaries of a physical campus environment. In many schools of architecture in Australia, the physical design studio learning environments however, remain largely unchanged. Many learning environments could be mistaken for those last refurbished 30 years ago, being devoid of any significant technological intervention. While some teaching staff are eagerly embracing new digital technologies and attempting to modify their pedagogical approaches, the physical design studio learning environment is resistant to such efforts. In a study aimed at better understanding how staff and students adapt to new blended learning environments, a group of 165 second year architecture students at a large school of architecture in Australia were separated into two different design studio learning environments. 70% of students were allocated to a traditional design studio setting and 30% to a new, high technology embedded, prototype digital learning laboratory. The digital learning laboratory was purpose designed for the case-study users, adapted Student-Centred Active Learning Environment for Undergraduate Programs [SCALE-UP] principles, and built as part of a larger university research project. The architecture students attended the same lectures, followed the same studio curriculum and completed the same pieces of assessment; the only major differences were the teaching staff and physical environment within which the studios were conducted. At the end of the semester, all staff and students were asked to complete a questionnaire about their experiences and preferences within the two respective learning environments. The questionnaire response rate represented the opinions of 100% of the 10 teaching staff and over 70% of the students. Using a qualitative grounded theory approach, data were coded, extrapolated and compared, to reveal emerging key themes. The key themes formed the basis for in-depth interviews and focus groups of teaching staff and students, allowing the researchers to understand the data in more detail. The results of the data verified what had become increasingly evident during the course of the semester: an underlying negative resistance to the new digital studio learning environment, by both staff and students. Many participants openly exhibited a yearning for a return to the traditional design studio learning environments, particularly when the new technology caused frustration, by being unreliable or failing altogether. This paper reports on the study, discusses the negative resistance and explores the major contributors to resistance. The researchers are not aware of any similar previous studies across these particular settings and believe that it offers a necessary and important contribution to emergent research about adaptation to new digital learning environments.

Improving the moment capacities of hollow flange cold-formed Litesteel Beams using web stiffeners

The LiteSteel Beam (LSB) is a new hollow flange section developed in Australia with a unique geometry consisting of torsionally rigid rectangular hollow flanges and a relatively slender web. The LSB is subjected to a relatively new Lateral Distortional Buckling (LDB) mode when used as flexural members. Unlike the commonly observed lateral torsional buckling, lateral distortional buckling of LSBs is characterised by cross sectional change due to web distortion. Lateral distortional buckling causes significant moment capacity reduction for LSBs with intermediate spans. Therefore a detailed investigation was undertaken to determine the methods of reducing the effects of lateral distortional buckling in LSB flexural members. For this purpose the use of web stiffeners was investigated using finite element analyses of LSBs with different web stiffener spacing and sizes. It was found that the use of 5 mm steel plate stiffeners welded or screwed to the inner faces of the top and bottom flanges at third span points considerably reduced the lateral distortional buckling effects in LSBs. Suitable design rules were then developed to calculate the enhanced elastic lateral distortional buckling moments and the higher ultimate moment capacities of LSBs with the chosen web stiffener arrangement. This paper presents the details of this investigation and the results.

Fourier Lucas-Kanade Algorithm

In this paper we propose a framework for both gradient descent image and object alignment in the Fourier domain. Our method centers upon the classical Lucas & Kanade (LK) algorithm where we represent the source and template/model in the complex 2D Fourier domain rather than in the spatial 2D domain. We refer to our approach as the Fourier LK (FLK) algorithm. The FLK formulation is advantageous when one pre-processes the source image and template/model with a bank of filters (e.g. oriented edges, Gabor, etc.) as: (i) it can handle substantial illumination variations, (ii) the inefficient pre-processing filter bank step can be subsumed within the FLK algorithm as a sparse diagonal weighting matrix, (iii) unlike traditional LK the computational cost is invariant to the number of filters and as a result far more efficient, and (iv) this approach can be extended to the inverse compositional form of the LK algorithm where nearly all steps (including Fourier transform and filter bank pre-processing) can be pre-computed leading to an extremely efficient and robust approach to gradient descent image matching. Further, these computational savings translate to non-rigid object alignment tasks that are considered extensions of the LK algorithm such as those found in Active Appearance Models (AAMs).

A coupled SPH/FEM analysis of portable water filled barriers

Portable water filled barriers (PWFB) are semi-rigid roadside barriers which have the potential to display good crash attenuation characteristics at low and moderate impact speeds. The traditional mesh based numerical methods alone fail to simulate this type of impact with precision, stability and efficiency. This paper proposes to develop an advanced simulation model based on the combination of Smoothed Particles Hydrodynamics (SPH), a meshless method, and finite element method (FEM) for fluid-structure analysis using the commercially available software package LS-Dyna. The interaction between SPH particles and FEA elements is studied in this paper. Two methods of element setup at the element boundary were investigated. The response of the impacted barrier and fluid inside were analysed and compared. The system response and lagging were observed and reported in this paper. It was demonstrated that coupled SPH/FEM can be used in full scale PWFB modelling application. This will aid the research in determining the best initial setup to couple FEA and SPH in road safety barrier for impact response and safety analysis in the future.

Temporal summation of intrinsically photosensitive retinal ganglion cell (ipRGC) contributions to the human pupil.

Purpose: IpRGCs mediate non-image forming functions including photoentrainment and the pupil light reflex (PLR). Temporal summation increases visual sensitivity and decreases temporal resolution for image forming vision, but the summation properties of nonimage forming vision are unknown. We investigated the temporal summation of inner (ipRGC) and outer (rod/cone) retinal inputs to the PLR. Method: The consensual PLR of the left eye was measured in six participants with normal vision using a Maxwellian view infrared pupillometer. Temporal summation was investigated using a double-pulse protocol (100 ms stimulus pairs; 0–1024 ms inter-stimulus interval, ISI) presented to the dilated fellow right eye (Tropicamide 1%). Stimulus lights (blue λmax = 460 nm; red λmax = 638 nm) biased activity to inneror outer retinal inputs to non-image forming vision. Temporal summation was measured suprathreshold (15.2 log photons.cm−2.s−1 at the cornea) and subthreshold (11.4 log photons.cm−2.s−1 at the cornea). Results: RM-ANOVAs showed the suprathreshold and subthreshold 6 second post illumination pupil response (PIPR: expressed as percentage baseline diameter) did not significantly vary for red or blue stimuli (p > .05). The PIPR for a subthreshold red 16 ms double-pulse control condition did not significantly differ with ISI (p > .05). The maximum constriction amplitude for red and blue 100 ms double- pulse stimuli did not significantly vary with ISI (p > .05). Conclusion: The non-significant changes in suprathreshold PIPR and subthreshold maximum pupil constriction indicate that inner retinal ipRGC inputs and outer retinal photoreceptor inputs to the PLR do not show temporal summation. The results suggest a fundamental difference between the temporal summation characteristics of image forming and non-image forming vision.