Second harmonic generation and multiphoton microscopic detection of collagen without the need for species specific antibodies

High-resolution, high-contrast, three-dimensional images of live cell and tissue architecture can be obtained using second harmonic generation (SHG), which comprises non-absorptive frequency changes in an excitation laser line. SHG does not require any exogenous antibody or fluorophore labeling, and can generate images from unstained sections of several key endogenous biomolecules, in a wide variety of species and from different types of processed tissue. Here, we examined normal control human skin sections and human burn scar tissues using SHG on a multi-photon microscope (MPM). Examination and comparison of normal human skin and burn scar tissue demonstrated a clear arrangement of fibers in the dermis, similar to dermal collagen fiber signals. Fluorescence-staining confirmed the MPM-SHG collagen colocalization with antibody staining for dermal collagen type-I but not fibronectin or elastin. Furthermore, we were able to detect collagen MPM-SHG signal in human frozen sections as well as in unstained paraffin embedded tissue sections that were then compared with hematoxylin and eosin staining in the identical sections. This same approach was also successful in localizing collagen in porcine and ovine skin samples, and may be particularly important when species-specific antibodies may not be available. Collectively, our results demonstrate that MPM SHG-detection is a useful tool for high resolution examination of collagen architecture in both normal and wounded human, porcine and ovine dermal tissue.

Innovation through action research in environmental education : from project to praxis

This thesis is a work-in-progress that articulates my research journey based on the development of a curriculum innovation in environmental education. This journey had two distinct, but intertwined phases: action research based fieldwork, conducted collaboratively, to create a whole school approach to environmental education curriculum planning; and a phase of analysis and reflection based on the emerging findings, as I sought to create personal "living educational theory" about change and innovation. A key stimulus for the study was the perceived theory-practice gap in environmental education, which is often presented in the literature as a criticism of teachers for failing to achieve the values and action objectives of critical environmental education. Hence, many programs and projects are considered to be superficial and inconsequential in terms of their ability to seriously address environmental issues. The intention of this study was to work with teachers in a project that would be an exemplar of critical environmental education. This would be in the form of a whole school "learnscaping" curriculum in a primary school whereby the schoolgrounds would be utilised for interdisciplinary critical environment education. Parallel with the three cycles of action research in this project, my research objectives were to identify and comment upon the factors that influence the generation of successful educational innovation. It was anticipated that the project would be a collaboration involving me, as researcher-facilitator, and many of the teachers in the school as active participants. As the project proceeded through its action cycles, however, it became obvious that the goal of developing a critical environmental education curriculum, and the use of highly participatory processes, were unrealistic. Institutional and organisational rigidities in education generally, teachers' day-to-day work demands, and the constant juggle of work, family and other responsibilities for all participants acted as significant constraints. Consequently, it became apparent that the learnscaping curriculum would not be the hoped-for exemplar. Progress was slow and, at times, the project was in danger of stalling permanently. While the curriculum had some elements of critical environmental education, these were minor and not well spread throughout the school. Overall, the outcome seemed best described as a "small win"; perhaps just another example of the theory-practice gap that I had hoped this project would bridge. Towards the project's end, however, my continuing reflection led to an exploration of chaos/complexity theory which gave new meaning to the concept of a "small win". According to this theory, change is not the product of linear processes applied methodically in purposeful and diligent ways, but emerges from serendipitous events that cannot be planned for, or forecast in advance. When this perspective of change is applied to human organisations - in this study, a busy school - the context for change is recognised not as a stable, predictable environment, but as a highly complex system where change happens all the time, cannot be controlled, and no one can be really sure where the impacts might lead. This so-called "butterfly effect" is a central idea of this theory where small changes or modifications are created - the effects of which are difficult to know, let alone determine - and which can have large-scale impacts. Allied with this effect is the belief that long term developments in an organisation that takes complexity into account, emerge by spontaneous self-organising evolution, requiring political interaction and learning in groups, rather than systematic progress towards predetermined goals or "visions". Hence, because change itself and the contexts of change are recognised as complex, chaos/complexity theory suggests that change is more likely to be slow and evolutionary - cultural change - rather than fast and revolutionary where the old is quickly ushered out by radical reforms and replaced by new structures and processes. Slow, small-scale changes are "normal", from a complexity viewpoint, while rapid, wholesale change is both unlikely and unrealistic. Therefore, the frustratingly slow, small-scale, imperfect educational changes that teachers create - including environmental education initiatives - should be seen for what they really are. They should be recognised as successful changes, the impacts of which cannot be known, but which have the potential to magnify into large-scale changes into the future. Rather than being regarded as failures for not meeting critical education criteria, "small wins" should be cause for celebration and support. The intertwined phases of collaborative action research and individual researcher reflection are mirrored in the thesis structure. The first three chapters, respectively, provide the thesis overview, the literature underpinning the study's central concern, and the research methodology. Chapters 4, 5, and 6 report on each of the three action research cycles of the study, namely Laying the Groundwork, Down to Work!, and The Never-ending Story. Each of these chapters presents a narrative of events, a literature review specific to developments in the cycle, and analysis and critique of the events, processes and outcomes of each cycle. Chapter 7 provides a synthesis of the whole of the study, outlining my interim propositions about facilitating curriculum change in schools through action research, and the implications of these for environmental education.

Controlled Monte Carlo data generation for statistical damage identification employing Mahalanobis squared distance

The use of Mahalanobis squared distance–based novelty detection in statistical damage identification has become increasingly popular in recent years. The merit of the Mahalanobis squared distance–based method is that it is simple and requires low computational effort to enable the use of a higher dimensional damage-sensitive feature, which is generally more sensitive to structural changes. Mahalanobis squared distance–based damage identification is also believed to be one of the most suitable methods for modern sensing systems such as wireless sensors. Although possessing such advantages, this method is rather strict with the input requirement as it assumes the training data to be multivariate normal, which is not always available particularly at an early monitoring stage. As a consequence, it may result in an ill-conditioned training model with erroneous novelty detection and damage identification outcomes. To date, there appears to be no study on how to systematically cope with such practical issues especially in the context of a statistical damage identification problem. To address this need, this article proposes a controlled data generation scheme, which is based upon the Monte Carlo simulation methodology with the addition of several controlling and evaluation tools to assess the condition of output data. By evaluating the convergence of the data condition indices, the proposed scheme is able to determine the optimal setups for the data generation process and subsequently avoid unnecessarily excessive data. The efficacy of this scheme is demonstrated via applications to a benchmark structure data in the field.

Numerical study of lid-driven square cavity with heat generation using LBM

In this study, the mixed convection heat transfer and fluid flow behaviors in a lid-driven square cavity filled with high Prandtl number fluid (Pr = 5400, ν = 1.2×10-4 m2/s) at low Reynolds number is studied using thermal Lattice Boltzmann method (TLBM) where ν is the viscosity of the fluid. The LBM has built up on the D2Q9 model and the single relaxation time method called the Lattice-BGK (Bhatnagar-Gross-Krook) model. The effects of the variations of non dimensional mixed convection parameter called Richardson number(Ri) with and without heat generating source on the thermal and flow behavior of the fluid inside the cavity are investigated. The results are presented as velocity and temperature profiles as well as stream function and temperature contours for Ri ranging from 0.1 to 5.0 with other controlling parameters that present in this study. It is found that LBM has good potential to simulate mixed convection heat transfer and fluid flow problem. Finally the simulation results have been compared with the previous numerical and experimental results and it is found to be in good agreement.

Contours in Motion

My practice-led research explores and maps workflows for generating experimental creative work involving inertia based motion capture technology. Motion capture has often been used as a way to bridge animation and dance resulting in abstracted visuals outcomes. In early works this process was largely done by rotoscoping, reference footage and mechanical forms of motion capture. With the evolution of technology, optical and inertial forms of motion capture are now more accessible and able to accurately capture a larger range of complex movements. The creative work titled “Contours in Motion” was the first in a series of studies on captured motion data used to generating experimental visual forms that reverberate in space and time. With the source or ‘seed’ comes from using an Xsens MVN - Inertial Motion Capture system to capture spontaneous dance movements, with the visual generation conducted through a customised dynamics simulation. The aim of the creative work was to diverge way from a standard practice of using particle system and/or a simple re-targeting of the motion data to drive a 3d character as a means to produce abstracted visual forms. To facilitate this divergence a virtual dynamic object was tether to a selection of data points from a captured performance. The proprieties of the dynamic object were then adjusted to balance the influences from the human movement data with the influence of computer based randomization. The resulting outcome was a visual form that surpassed simple data visualization to project the intent of the performer’s movements into a visual shape itself. The reported outcomes from this investigation have contributed to a larger study on the use of motion capture in the generative arts, furthering the understanding of and generating theories on practice.

Simulations of the spontaneous emission of a quantum dot near a gap plasmon waveguide

In this paper we modeled a quantum dot at near proximity to a gap plasmon waveguide to study the quantum dot-plasmon interactions. Assuming that the waveguide is single mode, this paper is concerned about the dependence of spontaneous emission rate of the quantum dot on waveguide dimensions such as width and height. We compare coupling efficiency of a gap waveguide with symmetric configuration and asymmetric configuration illustrating that symmetric waveguide has a better coupling efficiency to the quantum dot. We also demonstrate that optimally placed quantum dot near a symmetric waveguide with 50 nm x 50 nm cross section can capture 80% of the spontaneous emission into a guided plasmon mode.

Optimal distributed generation placement in radial distribution systems

Distributed generation (DG) resources are commonly used in the electric systems to obtain minimum line losses, as one of the benefits of DG, in radial distribution systems. Studies have shown the importance of appropriate selection of location and size of DGs. This paper proposes an analytical method for solving optimal distributed generation placement (ODGP) problem to minimize line losses in radial distribution systems using loss sensitivity factor (LSF) based on bus-injection to branch-current (BIBC) matrix. The proposed method is formulated and tested on 12 and 34 bus radial distribution systems. The classical grid search algorithm based on successive load flows is employed to validate the results. The main advantages of the proposed method as compared with the other conventional methods are the robustness and no need to calculate and invert large admittance or Jacobian matrices. Therefore, the simulation time and the amount of computer memory, required for processing data especially for the large systems, decreases.

Poly(2-oxazoline) hydrogels as next generation three dimensional cell supports

Synthetic hydrogels selectively decorated with cell adhesion motifs are rapidly emerging as promising substrates for 3D cell culture. When cells are grown in 3D they experience potentially more physiologically relevant cell-cell interactions and physical cues compared with traditional 2D cell culture on stiff surfaces. A newly developed polymer based on poly(2-oxazoline)s has been used for the first time to control attachment of fibroblast cells and is discussed here for its potential use in 3D cell culture with particular focus on cancer cells towards the ultimate aim of high throughput screening of anti-cancer therapies. Advantages and limitations of using poly(2-oxazoline) hydrogels are discussed and compared with more established polymers, especially polyethylene glycol (PEG).

Supplementary material : large scale read classification for next generation sequencing

Next Generation Sequencing (NGS) has revolutionised molecular biology, resulting in an explosion of data sets and an increasing role in clinical practice. Such applications necessarily require rapid identification of the organism as a prelude to annotation and further analysis. NGS data consist of a substantial number of short sequence reads, given context through downstream assembly and annotation, a process requiring reads consistent with the assumed species or species group. Highly accurate results have been obtained for restricted sets using SVM classifiers, but such methods are difficult to parallelise and success depends on careful attention to feature selection. This work examines the problem at very large scale, using a mix of synthetic and real data with a view to determining the overall structure of the problem and the effectiveness of parallel ensembles of simpler classifiers (principally random forests) in addressing the challenges of large scale genomics.

Increased generation of platelet-derived microparticles following percutaneous transluminal coronary angioplasty

Platelet-derived microparticles (PMPs) which are produced during platelet activation contribute to coagulation1 and bind to traumatized endothelium in an animal model2. Such endothelial injury occurs during percutaneous transluminal coronary angioplasty (PTCA), a procedure which restores the diameter of occluded coronary arteries using balloon inflations. However, re-occlusions subsequently develop in 20-25% of patients3, although this is limited by treatment with anti-platelet glycoprotein IIb/IIIa receptor drugs such as abciximab4. However, abciximab only partially decreases the need for revascularisation5, and therefore other mechanisms appear to be involved. As platelet activation occurs during PTCA, it is likely that PMPs may be produced and contribute to restenosis. This study population consisted of 113 PTCA patients, of whom 38 received abciximab. Paired peripheral arterial blood samples were obtained from the PTCA sheath: 1) following heparinisation (baseline); and 2) subsequent to all vessel manipulation (post-PTCA). Blood was prepared with an anti-CD61 (glycoprotein IIIa) fluorescence conjugated antibody to identify PMPs using flow cytometry, and PMP results expressed as a percentage of all CD61 events. The level of PMPs increased significantly from baseline following PTCA in the without abciximab group (paired t test, P=0.019). However, there was no significant change in the level of PMPs following PTCA in patients who received abciximab. Baseline clinical characteristics between patient groups were similar, although patients administered abciximab had more complex PTCA procedures, such as increased balloon inflation pressures (ANOVA, P=0.0219). In this study, we have clearly demonstrated that the level of CD61-positive PMPs increased during PTCA. This trend has been demonstrated previously, although a low sample size prevented statistical significance being attained6. The results of our work also demonstrate that there was no increase in PMPs after PTCA with abiciximab treatment. The increased PMPs may adhere to traumatized endothelium, contributing to re-occlusion of the arteries, but this remains to be determined. References: (1) Holme PA, Brosstad F, Solum NO. Blood Coagulation and Fibrinolysis. 1995;6:302-310. (2) Merten M, Pakala R, Thiagarajan P, Benedict CR. Circulation. 1999;99:2577-2582. (3) Califf RM. American Heart Journal.1995;130:680-684. (4) Coller BS, Scudder LE. Blood. 1985;66:1456-1459. (5) Topol EJ, Califf RM, Weisman HF, Ellis SG, Tcheng JE, Worley S, Ivanhoe R, George BS, Fintel D, Weston M, Sigmon K, Anderson KM, Lee KL, Willerson JT on behalf of the EPIC investigators. Lancet. 1994;343:881-886. (6) Scharf RE, Tomer A, Marzec UM, Teirstein PS, Ruggeri ZM, Harker LA. Arteriosclerosis and Thrombosis. 1992;12:1475-87.

Introduction to the special issue on next generation network architectures, protocols, theory, systems and applications

This special issue of Networking Science focuses on Next Generation Network (NGN) that enables the deployment of access independent services over converged fixed and mobile networks. NGN is a packet-based network and uses the Internet protocol (IP) to transport the various types of traffic (voice, video, data and signalling). NGN facilitates easy adoption of distributed computing applications by providing high speed connectivity in a converged networked environment. It also makes end user devices and applications highly intelligent and efficient by empowering them with programmability and remote configuration options. However, there are a number of important challenges in provisioning next generation network technologies in a converged communication environment. Some preliminary challenges include those that relate to QoS, switching and routing, management and control, and security which must be addressed on an urgent or emergency basis. The consideration of architectural issues in the design and pro- vision of secure services for NGN deserves special attention and hence is the main theme of this special issue.

Carbon, biodiversity and regional natural resource planning : towards high impact next generation plans

Given the increased importance of adaptation debates in global climate negotiations, pressure to achieve biodiversity, food and water security through managed landscape-scale adaptation will likely increase across the globe over the coming decade. In parallel, emerging market-based, terrestrial greenhouse gas abatement programs present a real opportunity to secure such adaptation to climate change through enhanced landscape resilience. Australia has an opportunity to take advantage of such programs through regional planning aspects of its governance arrangements for NRM. This paper explores necessary reforms to Australia's regional NRM planning systems to ensure that they will be better able to direct the nation's emerging GGA programs to secure enhanced landscape adaptation. © 2013 Planning Institute Australia.

Generation of two isomers of C5H from the corresponding anions. A theoretically motivated mass spectrometric study

Molecular orbital calculations have predicted the stability of a range of connectivities for the radical C5H potential surface. The most energetically favorable of these include the linear C4CH geometry and two ring-chain structures HC2C3 and C2C3H The corresponding anions are also shown to be theoretically stable, and furthermore, a fourth isomer, C2CHC2, is predicted to be the most stable anion connectivity. These results have motivated experimental efforts. Methodologies for the generation of the non-ring-containing isomeric anions C4CH and C2CHC2 have been developed utilizing negative ion mass spectrometry. The absolute connectivities of the anions have been established using deuterium labeling, charge reversal, and neutralization reionization techniques. The success of the latter experiment confirms theoretical predictions of stability of the corresponding neutral species. This is the first reported observation of the neutral C2CHC2 species that calculations predict to be substantially less stable than the C4CH connectivity but still bound relative to isomerization processes.

Generation and characterization of ionic and neutral dihydroxy boron B(OH)(2)(+/0) in the gas phase

The dicoordinated borinium ion, dihydroxyborinium, B(OH)(2)(+) is generated from methyl boronic acid CH3B(OH)(2) by dissociative electron ionization and its connectivity confirmed by collisional activation. Neutralization-reionization (NR) experiments on this ion indicate that the neutral B(OH)(2) radical is a viable species in the gas phase. Both vertical neutralization of B(OH)(2)(+) and reionization of B(OH)(2) in the NR experiment are, however, associated with particularly unfavorable Franck-Condon factors. The differences in adiabatic and vertical electron transfer behavior can be traced back to a particular pi stabilization of the cationic species compared to the sp(2)-type neutral radical. Thermochemical data on several neutral and cationic boron compounds are presented based on calculations performed at the G2 level of theory.

Aligning in and through interaction : children getting in and out of spontaneous activity

Spontaneous play, important for forming the basis of friendships and peer relations, is a complex activity involving the management and production of talk-in-interaction. This paper focuses on the intricacies of social interaction, emphasising the link between alignment and affiliation, and the range and importance of verbal and nonverbal interactive devices available to children. Analysis of the way in which two girls, one of whom has been diagnosed with Asperger’s Syndrome, engage in spontaneous activities demonstrates the potential for interactional difficulty due to the unscripted nature of the interaction. The paper argues for further research into how improvised, unscripted interactions are initiated within moment-by-moment talk, how they unfold, and how they are brought to a close in everyday contexts in order to understand how children create their social worlds.