Large-volume methacrylate monolith for plasmid purification : process engineering approach to synthesis and application

The extent of exothermicity associated with the construction of large-volume methacrylate monolithic columns has somewhat obstructed the realisation of large-scale rapid biomolecule purification especially for plasmid-based products which have proven to herald future trends in biotechnology. A novel synthesis technique via a heat expulsion mechanism was employed to prepare a 40 mL methacrylate monolith with a homogeneous radial pore structure along its thickness. Radial temperature gradient was recorded to be only 1.8 °C. Maximum radial temperature recorded at the centre of the monolith was 62.3 °C, which was only 2.3 °C higher than the actual polymerisation temperature. Pore characterisation of the monolithic polymer showed unimodal pore size distributions at different radial positions with an identical modal pore size of 400 nm. Chromatographic characterisation of the polymer after functionalisation with amino groups displayed a persistent dynamic binding capacity of 15.5 mg of plasmid DNA/mL. The maximum pressure drop recorded was only 0.12 MPa at a flow rate of 10 mL/min. The polymer demonstrated rapid separation ability by fractionating Escherichia coli DH5α-pUC19 clarified lysate in only 3 min after loading. The plasmid sample collected after the fast purification process was tested to be a homogeneous supercoiled plasmid with DNA electrophoresis and restriction analysis.

Process engineering approach to large-volume methacrylate monolith synthesis for plasmid purification

The construction of large?volume methacrylate monolithic columns for preparative-scale plasmid purification is obstructed by the enormous release of exotherms, thus introducing structural heterogeneity in the monolith pore system. A remarkable radial temperature gradient develops along the monolith thickness, reaching a terminal temperature that supersedes the maximum temperature required for the preparation of a structurally homogeneous monolith. A novel heat expulsion technique is employed to overcome the heat build-up during the synthesis process. The enormous heat build-up is perceived to encompass the heat associated with initiator decomposition and the heat released from free radical-monomer and monomer-monomer interactions. The heat resulting from the initiator decomposition was expelled along with some gaseous fumes before commencing polymerisation in a gradual addition fashion. Characteristics of a 50 mL monolith synthesized using this technique showed an improved uniformity in the pore structure radially along the length on the monolith. Chromatographic characterization of this adsorbent displayed a persistent binding capacity of 14.5 mg pDNA/mL of the adsorbent. The adsorbent was able to fractionate a clarified bacteria lysate in only 3 min (after loading) into RNA, protein and pDNA respectively. The pDNA fraction obtained was analyzed to be a homogeneous supercoiled pDNA.

Large-scale habitat mapping using vision-based AUVs : experiences, challenges & vehicle design

This paper overviews the development of a vision-based AUV along with a set of complementary operational strategies to allow reliable autonomous data collection in relatively shallow water and coral reef environments. The development of the AUV, called Starbug, encountered many challenges in terms of vehicle design, navigation and control. Some of these challenges are discussed with focus on operational strategies for estimating and reducing the total navigation error when using lower-resolution sensing modalities. Results are presented from recent field trials which illustrate the ability of the vehicle and associated operational strategies to enable rapid collection of visual data sets suitable for marine research applications.

The origin of cyclical high- and low-Cr basalts with continental crust trace element signatures in the Kalkarindji Large Igneous Province

The 510 million year old Kalkarindji Large Igneous Province correlates in time with the first major extinction event after the Cambrian explosion of life. Large igneous provinces correlate with all major mass extinction events in the last 500 million years. The genetic link between large igneous provinces and mass extinction remains unclear. My work is a contribution towards understanding magmatic processes involved in the generation of Large Igneous Provinces. I concentrate on the origin of variation in Cr in magmas and have developed a model in which high temperature melts intrude into and assimilate large amounts of upper continental crust.

Vision-based operations of a large industrial vehicle : autonomous hot metal carrier

Hot metal carriers (HMCs) are large forklift-type vehicles used to move molten metal in aluminum smelters. This paper reports on field experiments that demonstrate that HMCs can operate autonomously and in particular can use vision as a primary sensor to locate the load of aluminum. We present our complete system but focus on the vision system elements and also detail experiments demonstrating reliable operation of the materials handling task. Two key experiments are described, lasting 2 and 5 h, in which the HMC traveled 15 km in total and handled the load 80 times.

Histogram matching and global initialization for laser-only SLAM in large unstructured environments

This paper presents an enhanced algorithm for matching laser scan maps using histogram correlations. The histogram representation effectively summarizes a map's salient features such that pairs of maps can be matched efficiently without any prior guess as to their alignment. The histogram matching algorithm has been enhanced in order to work well in outdoor unstructured environments by using entropy metrics, weighted histograms and proper thresholding of quality metrics. Thus our large-scale scan-matching SLAM implementation has a vastly improved ability to close large loops in real-time even when odometry is not available. Our experimental results have demonstrated a successful mapping of the largest area ever mapped to date using only a single laser scanner. We also demonstrate our ability to solve the lost robot problem by localizing a robot to a previously built map without any prior initialization.

Dynamic agent composition for large-scale agent-based models

PURPOSE: This paper describes dynamic agent composition, used to support the development of flexible and extensible large-scale agent-based models (ABMs). This approach was motivated by a need to extend and modify, with ease, an ABM with an underlying networked structure as more information becomes available. Flexibility was also sought after so that simulations are set up with ease, without the need to program. METHODS: The dynamic agent composition approach consists in having agents, whose implementation has been broken into atomic units, come together at runtime to form the complex system representation on which simulations are run. These components capture information at a fine level of detail and provide a vast range of combinations and options for a modeller to create ABMs. RESULTS: A description of the dynamic agent composition is given in this paper, as well as details about its implementation within MODAM (MODular Agent-based Model), a software framework which is applied to the planning of the electricity distribution network. Illustrations of the implementation of the dynamic agent composition are consequently given for that domain throughout the paper. It is however expected that this approach will be beneficial to other problem domains, especially those with a networked structure, such as water or gas networks. CONCLUSIONS: Dynamic agent composition has many advantages over the way agent-based models are traditionally built for the users, the developers, as well as for agent-based modelling as a scientific approach. Developers can extend the model without the need to access or modify previously written code; they can develop groups of entities independently and add them to those already defined to extend the model. Users can mix-and-match already implemented components to form large-scales ABMs, allowing them to quickly setup simulations and easily compare scenarios without the need to program. The dynamic agent composition provides a natural simulation space over which ABMs of networked structures are represented, facilitating their implementation; and verification and validation of models is facilitated by quickly setting up alternative simulations.

Automotive modal lock-in: The role of path dependence and large socio-economic regimes in market failure

This paper addresses less recognised factors which influence the diffusion of a particular technology. While an innovation’s attributes and performance are paramount, many fail because of external factors which favour an alternative. This paper, with theoretic input from diffusion, lock-in and path-dependency, presents a qualitative study of external factors that influenced the evolution of transportation in USA. This historical account reveals how one technology and its emergent systems become dominant while other choices are overridden by socio-political, economic and technological interests which include not just the manufacturing and service industries associated with the automobile but also government and market stakeholders. Termed here as a large socio-economic regime (LSER),its power in ensuring lock-in and continued path-dependency is shown to pass through three stages, weakening eventually as awareness improves. The study extends to transport trends in China, Korea, Indonesia and Malaysia and they all show the dominant role of an LSER. As transportation policy is increasingly accountable to address both demand and environmental concerns and innovators search for solutions, this paper presents important knowledge for innovators, marketers and policy makers for commercial and societal reasons, especially when negative externalities associated with an incumbent transportation technology may lead to market failure.

Large-scale interlaboratory study to develop, analytically validate and apply highly multiplexed, quantitative peptide assays to measure cancer-relevant proteins in plasma

There is an increasing need in biology and clinical medicine to robustly and reliably measure tens-to-hundreds of peptides and proteins in clinical and biological samples with high sensitivity, specificity, reproducibility and repeatability. Previously, we demonstrated that LC-MRM-MS with isotope dilution has suitable performance for quantitative measurements of small numbers of relatively abundant proteins in human plasma, and that the resulting assays can be transferred across laboratories while maintaining high reproducibility and quantitative precision. Here we significantly extend that earlier work, demonstrating that 11 laboratories using 14 LC-MS systems can develop, determine analytical figures of merit, and apply highly multiplexed MRM-MS assays targeting 125 peptides derived from 27 cancer-relevant proteins and 7 control proteins to precisely and reproducibly measure the analytes in human plasma. To ensure consistent generation of high quality data we incorporated a system suitability protocol (SSP) into our experimental design. The SSP enabled real-time monitoring of LC-MRM-MS performance during assay development and implementation, facilitating early detection and correction of chromatographic and instrumental problems. Low to sub-nanogram/mL sensitivity for proteins in plasma was achieved by one-step immunoaffinity depletion of 14 abundant plasma proteins prior to analysis. Median intra- and inter-laboratory reproducibility was <20%, sufficient for most biological studies and candidate protein biomarker verification. Digestion recovery of peptides was assessed and quantitative accuracy improved using heavy isotope labeled versions of the proteins as internal standards. Using the highly multiplexed assay, participating laboratories were able to precisely and reproducibly determine the levels of a series of analytes in blinded samples used to simulate an inter-laboratory clinical study of patient samples. Our study further establishes that LC-MRM-MS using stable isotope dilution, with appropriate attention to analytical validation and appropriate quality c`ontrol measures, enables sensitive, specific, reproducible and quantitative measurements of proteins and peptides in complex biological matrices such as plasma.

Experiments and experiences with dependability for a large autonomous industrial vehicle

This paper describes the experiences gained performing multiple experiments while developing a large autonomous industrial vehicle. Hot Metal Carriers (HMCs) are large forklift-type vehicles used in the light metals industry to move molten or hot metal around a smelter. Autonomous vehicles of this type must be dependable as they are large and potentially hazardous to infrastructure and people. This paper will talk about four aspects of dependability, that of safety, reliability, availability and security and how they have been addressed on our experimental autonomous HMC.

Analysis of moderation practices in a large STEM-focused faculty

Moderation of assessment constitutes a crucial element of the learning and teaching process at the university. Yet, despite its importance, many academics have confusing beliefs and attitudes towards moderation practices, processes and procedures. This paper reports on a qualitative study conducted in a Science, Technology, Engineering and Mathematics (STEM)-focused faculty at a large Australian higher education institution. The findings of the study revealed a strong need for further investigation on the ways moderation is understood and enacted by academics within a STEM-specific context and informed redevelopment of the faculty’s internal moderation policy.

MRI reveals individual level deformities in the growing scoliotic spine that clinically are masked by the Cobb angle

Clinically, the Cobb angle method measures the overall scoliotic curve in the coronal plane but does not measure individual vertebra and disc wedging. The contributions of the vertebrae and discs in the growing scoliotic spine were measured using sequential MRI scans to investigate coronal plane deformity progression with growth. Sequential MRI data showed complex patterns of deformity progression. Changes to the wedging of individual vertebrae and discs may occur in patients who have no increase in overall Cobb angle measure; the Cobb method alone may be insufficient to capture the complex mechanisms of deformity progression.

Cyclic Salinity Model for deposition of large-scale coal deposits

Existing field data for Rangal coals (Late Permian) of the Bowen Basin, Queensland, Australia, are inconsistent with the depositional model generally accepted in the current geological literature to explain coal deposition. Given the apparent unsuitability of the current depositional model to the Bowen Basin coal data, a new depositional model, here named the Cyclic Salinity Model, is proposed and tested in this study.

The origin of a large (>3km) maar volcano by coalescence of multiple shallow craters: Lake Purrumbete maar, southeastern Australia

Lake Purrumbete maar is located in the intraplate, monogenetic Newer Volcanics Province in southeastern Australia. The extremely large crater of 3000. m in diameter formed on an intersection of two fault lines and comprises at least three coalesced vents. The evolution of these vents is controlled by the interaction of the tectonic setting and the properties of both hard and soft rock aquifers. Lithics in the maar deposits originate from country rock formations less than 300. m deep, indicating that the large size of the crater cannot only be the result of the downwards migration of the explosion foci in a single vent. Vertical crater walls and primary inward dipping beds evidence that the original size of the crater has been largely preserved. Detailed mapping of the facies distributions, the direction of transport of base surges and pyroclastic flows, and the distribution of ballistic block fields, form the basis for the reconstruction of the complex eruption history,which is characterised by alternations of the eruption style between relatively dry and wet phreatomagmatic conditions, and migration of the vent location along tectonic structures. Three temporally separated eruption phases are recognised, each starting at the same crater located directly at the intersection of two local fault lines. Activity then moved quickly to different locations. A significant volcanic hiatus between two of the three phases shows that the magmatic system was reactivated. The enlargement of especially the main crater by both lateral and vertical growth led to the interception of the individual craters and the formation of the large circular crater. Lake Purrumbete maar is an excellent example of how complicated the evolution of large, seemingly simple, circular maar volcanoes can be, and raises the question if these systems are actually monogenetic.