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.

Absence of intersystem crossing in 1,4-didehydrobenzene

The rate of singlet-to-triplet intersystem crossing in 1,4-didehydrobenzene (the biradical produced as a reactive intermediate in the thermal cycloaromatization of enediynes), cannot be increased by the application of an external magnetic field. The rate of product formation and the distribution of stable products of 2,3-di-n-propyl-1,4-didehydrobenzene thermolysis is unchanged at magnetic flux densities in the range 0–2000 G and at 66 000 G. Similarly, the rate of thermolysis of an unsymmetrical enediyne is insensitive to magnetic field flux in the same range. This finding precludes the modulation of enediyne reaction rates in pharmaceutical and synthetic pursuits.

Evaluation of the iPhone with an acrylic sleeve versus the Scoliometer for rib hump measurement in scoliosis

Background. Vertebral rotation found in structural scoliosis contributes to trunkal asymmetry which is commonly measured with a simple Scoliometer device on a patient's thorax in the forward flexed position. The new generation of mobile 'smartphones' have an integrated accelerometer, making accurate angle measurement possible, which provides a potentially useful clinical tool for assessing rib hump deformity. This study aimed to compare rib hump angle measurements performed using a Smartphone and traditional Scoliometer on a set of plaster torsos representing the range of torsional deformities seen in clinical practice. Methods. Nine observers measured the rib hump found on eight plaster torsos moulded from scoliosis patients with both a Scoliometer and an Apple iPhone on separate occasions. Each observer repeated the measurements at least a week after the original measurements, and were blinded to previous results. Intra-observer reliability and inter-observer reliability were analysed using the method of Bland and Altman and 95% confidence intervals were calculated. The Intra-Class Correlation Coefficients (ICC) were calculated for repeated measurements of each of the eight plaster torso moulds by the nine observers. Results. Mean absolute difference between pairs of iPhone/Scoliometer measurements was 2.1 degrees, with a small (1 degrees) bias toward higher rib hump angles with the iPhone. 95% confidence intervals for intra-observer variability were +/- 1.8 degrees (Scoliometer) and +/- 3.2 degrees (iPhone). 95% confidence intervals for inter-observer variability were +/- 4.9 degrees (iPhone) and +/- 3.8 degrees (Scoliometer). The measurement errors and confidence intervals found were similar to or better than the range of previously published thoracic rib hump measurement studies. Conclusions. The iPhone is a clinically equivalent rib hump measurement tool to the Scoliometer in spinal deformity patients. The novel use of plaster torsos as rib hump models avoids the variables of patient fatigue and discomfort, inconsistent positioning and deformity progression using human subjects in a single or multiple measurement sessions.

Integrity, attribution, and exploitation : contractual and normative moral rights protection in open licensing

Over the last twenty years, the use of open content licenses has become increasingly and surprisingly popular. The use of such licences challenges the traditional incentive-based model of exclusive rights under copyright. Instead of providing a means to charge for the use of particular works, what seems important is mitigating against potential personal harm to the author and, in some cases, preventing non-consensual commercial exploitation. It is interesting in this context to observe the primacy of what are essentially moral rights over the exclusionary economic rights. The core elements of common open content licences map somewhat closely to continental conceptions of the moral rights of authorship. Most obviously, almost all free software and free culture licences require attribution of authorship. More interestingly, there is a tension between social norms developed in free software communities and those that have emerged in the creative arts over integrity and commercial exploitation. For programmers interested in free software, licence terms that prohibit commercial use or modification are almost completely inconsistent with the ideological and utilitarian values that underpin the movement. For those in the creative industries, on the other hand, non-commercial terms and, to a lesser extent, terms that prohibit all but verbatim distribution continue to play an extremely important role in the sharing of copyright material. While prohibitions on commercial use often serve an economic imperative, there is also a certain personal interest for many creators in avoiding harmful exploitation of their expression – an interest that has sometimes been recognised as forming a component of the moral right of integrity. One particular continental moral right – the right of withdrawal – is present neither in Australian law or in any of the common open content licences. Despite some marked differences, both free software and free culture participants are using contractual methods to articulate the norms of permissible sharing. Legal enforcement is rare and often prohibitively expensive, and the various communities accordingly rely upon shared understandings of acceptable behaviour. The licences that are commonly used represent a formalised expression of these community norms and provide the theoretically enforceable legal baseline that lends them legitimacy. The core terms of these licences are designed primarily to alleviate risk in sharing and minimise transaction costs in sharing and using copyright expression. Importantly, however, the range of available licences reflect different optional balances in the norms of creating and sharing material. Generally, it is possible to see that, stemming particularly from the US, open content licences are fundamentally important in providing a set of normatively accepted copyright balances that reflect the interests sought to be protected through moral rights regimes. As the cost of creation, distribution, storage, and processing of expression continues to fall towards zero, there are increasing incentives to adopt open content licences to facilitate wide distribution and reuse of creative expression. Thinking of these protocols not only as reducing transaction costs but of setting normative principles of participation assists in conceptualising the role of open content licences and the continuing tensions that permeate modern copyright law.

The development of ground impact models for the analysis of the risks associated with Unmanned Aircraft Operations over inhabited areas

With the emergence of Unmanned Aircraft Systems (UAS) there is a growing need for safety standards and regulatory frameworks to manage the risks associated with their operations. The primary driver for airworthiness regulations (i.e., those governing the design, manufacture, maintenance and operation of UAS) are the risks presented to people in the regions overflown by the aircraft. Models characterising the nature of these risks are needed to inform the development of airworthiness regulations. The output from these models should include measures of the collective, individual and societal risk. A brief review of these measures is provided. Based on the review, it was determined that the model of the operation of an UAS over inhabited areas must be capable of describing the distribution of possible impact locations, given a failure at a particular point in the flight plan. Existing models either do not take the impact distribution into consideration, or propose complex and computationally expensive methods for its calculation. A computationally efficient approach for estimating the boundary (and in turn area) of the impact distribution for fixed wing unmanned aircraft is proposed. A series of geometric templates that approximate the impact distributions are derived using an empirical analysis of the results obtained from a 6-Degree of Freedom (6DoF) simulation. The impact distributions can be aggregated to provide impact footprint distributions for a range of generic phases of flight and missions. The maximum impact footprint areas obtained from the geometric template are shown to have a relative error of typically less than 1% compared to the areas calculated using the computationally more expensive 6DoF simulation. Computation times for the geometric models are on the order of one second or less, using a standard desktop computer. Future work includes characterising the distribution of impact locations within the footprint boundaries.

Excess power circulation in distribution networks containing distributed energy resources

The presence of large number of single-phase distributed energy resources (DERs) can cause severe power quality problems in distribution networks. The DERs can be installed in random locations. This may cause the generation in a particular phase exceeds the load demand in that phase. Therefore the excess power in that phase will be fed back to the transmission network. To avoid this problem, the paper proposes the use of distribution static compensator (DSTATCOM) that needs to be connected at the first bus following a substation. When operated properly, the DSTATCOM can facilitate a set of balanced current flow from the substation, even when excess power is generated by DERs. The proposals are validated through extensive digital computer simulation studies using PSCAD and MATLAB.

An approach for current balancing in distribution networks with rooftop PVs

Current unbalance is a significant power quality problem in distribution networks. This problem increases further with the increased penetration of single-phase photovoltaic cells. In this paper, a new approach is developed for current unbalance reduction in medium voltage distribution networks. The method is based on utilization of three single-phase voltage source converters connected in delta configuration between the phases. Each converter is controlled to function as a varying capacitor. The combination of the load and the compensator will result in a balanced load with unity power factor. The efficacy of the proposed current unbalance reduction concept is verified through dynamic simulations in PSCAD/EMTDC.

Distribution feeder loads classification and decomposition

Load in distribution networks is normally measured at the 11kV supply points; little or no information is known about the type of customers and their contributions to the load. This paper proposes statistical methods to decompose an unknown distribution feeder load to its customer load sector/subsector profiles. The approach used in this paper should assist electricity suppliers in economic load management, strategic planning and future network reinforcements.

How important is length? : mechanical testing and measurement of a cemented, polished, tapered femoral implant

Total hip arthroplasty (THA) has a proven clinical record for providing pain relief and return of function to patients with disabling arthritis. There are many successful options for femoral implant design and fixation. Cemented, polished, tapered femoral implants have been shown to have excellent results in national joint registries and long-term clinical series. These implants are usually 150mm long at their lateral aspect. Due to their length, these implants cannot always be offered to patients due to variations in femoral anatomy. Polished, tapered implants as short as 95mm exist, however their small proximal geometry (neck offset and body size) limit their use to smaller stature patients. There is a group of patients in which a shorter implant with a maintained proximal body size would be advantageous. There are also potential benefits to a shorter implant in standard patient populations such as reduced bone removal due to reduced reaming, favourable loading of the proximal femur, and the ability to revise into good proximal bone stock if required. These factors potentially make a shorter implant an option for all patient populations. The role of implant length in determining the stability of a cemented, polished, tapered femoral implant is not well defined by the literature. Before changes in implant design can be made, a better understanding of the role of each region in determining performance is required. The aim of the thesis was to describe how implant length affects the stability of a cemented, polished, tapered femoral implant. This has been determined through an extensive body of laboratory testing. The major findings are that for a given proximal body size, a reduction in implant length has no effect on the torsional stability of a polished, tapered design, while a small reduction in axial stability should be expected. These findings are important because the literature suggests that torsional stability is the major determinant of long-term clinical performance of a THA system. Furthermore, a polished, tapered design is known to be forgiving of cement-implant interface micromotion due to the favourable wear characteristics. Together these findings suggest that a shorter polished, tapered implant may be well tolerated. The effect of a change in implant length on the geometric characteristics of polished, tapered design were also determined and applied to the mechanical testing. Importantly, interface area does play a role in stability of the system; however it is the distribution of the interface and not the magnitude of the area that defines stability. Taper angle (at least in the range of angles seen in this work) was shown not to be a determinant of axial or torsional stability. A range of implants were tested, comparing variations in length, neck offset and indication (primary versus cement-in-cement revision). At their manufactured length, the 125mm implants were similar to their longer 150mm counterparts suggesting that they may be similarly well tolerated in the clinical environment. However, the slimmer cement-in-cement revision implant was shown to have a poorer mechanical performance, suggesting their use in higher demand patients may be hazardous. An implant length of 125mm has been shown to be quite stable and the results suggest that a further reduction to 100mm may be tolerated. However, further work is required. A shorter implant with maintained proximal body size would be useful for the group of patients who are unable to access the current standard length implants due to variations in femoral anatomy. Extending the findings further, the similar function with potential benefits of a shorter implant make their application to all patients appealing.

Evaluation of precision time synchronisation methods for substation applications

Many substation applications require accurate time-stamping. The performance of systems such as Network Time Protocol (NTP), IRIG-B and one pulse per second (1-PPS) have been sufficient to date. However, new applications, including IEC 61850-9-2 process bus and phasor measurement, require accuracy of one microsecond or better. Furthermore, process bus applications are taking time synchronisation out into high voltage switchyards where cable lengths may have an impact on timing accuracy. IEEE Std 1588, Precision Time Protocol (PTP), is the means preferred by the smart grid standardisation roadmaps (from both the IEC and US National Institute of Standards and Technology) of achieving this higher level of performance, and integrates well into Ethernet based substation automation systems. Significant benefits of PTP include automatic path length compensation, support for redundant time sources and the cabling efficiency of a shared network. This paper benchmarks the performance of established IRIG-B and 1-PPS synchronisation methods over a range of path lengths representative of a transmission substation. The performance of PTP using the same distribution system is then evaluated and compared to the existing methods to determine if the performance justifies the additional complexity. Experimental results show that a PTP timing system maintains the synchronising performance of 1-PPS and IRIG-B timing systems, when using the same fibre optic cables, and further meets the needs of process buses in large substations.

The spatial distribution of rodent damage in Australian macadamia (Macadamia integrifolia) orchards

The Black Rat (Rattus rattus), a global pest within the macadamia production industry, causes up to 30% crop damage in Australian orchards. During early stages of production in Australia, research demonstrated the importance of non crop adjacent habitats as significant in affecting the patterns of crop damage seen throughout orchards. Where once rodent damage was limited to the outside edges of orchard blocks, growers are now reporting finding crop damage throughout entire orchards. This study therefore aims to explore the spatial patterns of rodent distribution and damage now occurring in Australian macadamia orchards. We show that rodent damage and rodent distribution in these newer production regions differ from that shown in previous Australian research. Previous Australian research has shown damage patterns which were associated with the edges of orchard blocks however this study demonstrates a more widespread damage distribution. In the current study there is no relationship between rodent damage and the orchard edge. Arboreal rodent nests were identified within these newer orchard systems, suggesting rodents are residing within the tree component of the orchard system and not dependent on adjacent non-crop habitat for shelter. Results from this study confirm that rodents have modified their nesting and foraging behaviour in newer orchards systems in Australia. We suggest that this is a response of increased and prolonged availability of macadamia nuts in newer production regions enabling populations to be maintained throughout the year. Management strategies will require modification if control is to be achieved.

Restructuring of carbonaceous particles upon exposure to organic vapours

Recent research has described the restructuring of particles upon exposure to organic vapours; however, as yet hypotheses able to explain this phenomenon are limited. In this study, a range of experiments were performed to explore different hypotheses related to carbonaceous particle restructuring upon exposure to organic and water vapours, such as: the effect of surface tension, the role of organics in flocculating primary particles, as well as the ability of vapours to “wet” the particle surface. The change in mobility diameter (dm) was investigated for a range carbonaceous particle types (diesel exhaust, petrol exhaust, cigarette smoke, candle smoke, particles generated in a heptane/toluene flame, and wood smoke particles) exposed to different organic (heptane, ethanol, and dimethyl sulfoxide/water (1:1 vol%) mixture) and water vapours. Particles were first size-selected and then bubbled through an impinger (bubbler) containing either an organic solvent or water, where particles trapped inside rising bubbles were exposed to saturated vapours of the solvent in the impinger. The size distribution of particles was simultaneously measured upstream and downstream from the impinger. A size-dependent reduction in dm was observed when bubbling diesel exhaust, particles generated in a heptane/toluene flame, and candle smoke particles through heptane, ethanol and a dimethyl sulfoxide/water (1:1 vol %) mixture. In addition, the size distributions of particles bubbled through an impinger were broader. Moreover, an increase of the geometric standard deviation (σ) of the size distributions of particles bubbled through an impinger was also found to be size-dependent. Size-dependent reduction in dm and an increase of σ indicate that particles undergo restructuring to a more compact form, which was confirmed by TEM analysis. However, bubbling of these particles through water did not result in a size-dependent reduction in dm, nor in an increase of σ. Cigarette smoke, petrol exhaust, and wood smoke particles did not result in any substantial change in dm, or σ, when bubbled through organic solvents or water. Therefore, size-dependent reduction in the dm upon bubbling through organic solvents was observed only for particles that had a fractal-like structure, whilst particles that were liquid or were assumed to be spherical did not exhibit any reduction in dm. Compaction of fractal-like particles was attributed to the ability of condensing vapours to efficiently wet the particles. Our results also show that the presence of an organic layer on the surface of fractal-like particles, or the surface tension of the condensed liquid do not influence the extent of compaction.