A high performance liquid chromatography and electrospray ionization mass spectrometry method for the analysis of the natural medicine, Forsythia suspensa

Samples of Forsythia suspensa from raw (Laoqiao) and ripe (Qingqiao) fruit were analyzed with the use of HPLC-DAD and the EIS-MS techniques. Seventeen peaks were detected, and of these, twelve were identified. Most were related to the glucopyranoside molecular fragment. Samples collected from three geographical areas (Shanxi, Henan and Shandong Provinces), were discriminated with the use of hierarchical clustering analysis (HCA), discriminant analysis (DA), and principal component analysis (PCA) models, but only PCA was able to provide further information about the relationships between objects and loadings; eight peaks were related to the provinces of sample origin. The supervised classification models-K-nearest neighbor (KNN), least squares support vector machines (LS-SVM), and counter propagation artificial neural network (CP-ANN) methods, indicated successful classification but KNN produced 100% classification rate. Thus, the fruit were discriminated on the basis of their places of origin.

Profiled steel roof claddings under high wind forces

Profiled steel roof claddings in Australia and its neighbouring countries are commonly made of very thin high tensile steel and are crest-fixed intermittently with screw fasteners. The failure of the roof cladding systems was due to a local failure (dimpling of crests I pull-through) at the fasteners under wind uplift Cyclic wind uplift during cyclones causes fatigue cracking to occur at the fastener holes which leads to pull-through failures at lower load levels. At present the design of these claddings is entirely based on testing. In order to improve the understanding of the behaviour and the design and test methods of these claddings under wind uplift loading during storms and cyclones, a detailed investigation consisting of finite element analyses, static and fatigue experiments and cyclonic wind modelling was carried out on two-span roofing assemblies of three common roofing profiles. This paper presents the details of this investigation and its important results.

High-resolution GNSS-tracked drifter for studying surface dispersion in shallow water

The use of GNSS tracked Lagrangian drifters allows more realistic quantification of fluid motion and dispersion coefficients than Eulerian techniques because such drifters are analogues of particles that are relevant to flow field characterisation and pollutant dispersion. Using the fast growing Real Time Kinematic (RTK) positioning technique derived from Global Satellite Navigation Systems (GNSS), drifters are developed for high frequency (10 Hz) sampling with position estimates to centimetre accuracy. The drifters are designed with small size and less direct wind drag to follow the sub-surface flow which characterizes dispersion in shallow waters. An analysis of position error from stationary observation indicates that the drifter can efficiently resolve motion up to 1 Hz. The result of the field deployments of the drifter in conjunction with acoustic Eulerian devices shows higher estimate of the drifter streamwise velocities. Single particle statistical analysis of field deployments in a shallow estuarine zone yielded dispersion coefficients estimate comparable to those of dye tracer studies. The drifters capture the tidal elevation during field studies in a tidal estuary.

Afterschool MediaClub: Critical literacy in a high diversity, high poverty urban setting

In this chapter we present analyses of data produced with young people in an afterschool digital literacy program for 9 – 12 year olds. The young people were students at a high diversity, high poverty outer suburban elementary school in Queensland, Australia. The club was part of the URLearning research project (2010-14). In the classroom-based component of the project we worked with teachers to develop intellectually substantive and critical digital literacy practice. MediaClub was in some ways complementary to the classroom component; it was designed to skill up interested kids as digital media experts not only for their families and communities, but also for the classroom. Given the critical literacy traditions established in Australian schools, we approached MediaClub with certain critical expectations. In this chapter we look at what ensued, highlighting unanticipated critical outcomes at a time of heightened struggle over English curriculum. Critical literacy has been part of official English curriculum in Queensland since the early 1990s. The approach has been primarily text analytic, concerned with giving students access to genres of power and tools for understanding the ideological work of language through text. Many ideas for translating this normative critical project into classroom practice have been developed for use from the earliest elementary grades onwards. However, curricular space for critical literacy is under pressure. Amongst other things, this reflects both the development of Australia’s first national curriculum and the construction of a regimen of national literacy testing. At MediaClub we found a certain resistance to learning activities which were “too much like school”. However, in a context of increased control of teachers’ and students’ work in the classroom, MediaClub evolved as a learning space that can be understood in critical terms. Our experience in this regard might be of interest to teachers and researchers in high diversity high poverty settings that are strongly controlled through increasingly prescriptive – even scripted – pedagogies.

Microparticle-mediated gene delivery for the enhanced expression of a 19-KDa fragment of merozoite surface protein 1 of Plasmodium falciparum

The 19 kDa carboxyl-terminal fragment of merozoite surface protein 1 (MSP119) is a major component of the invasion-inhibitory response in individual immunity to malaria. A novel ultrasonic atomization approach for the formulation of biodegradable poly(lactic-co-glycolic acid) (PLGA) microparticles of malaria DNA vaccines encoding MSP119 is presented here. After condensing the plasmid DNA (pDNA) molecules with a cationic polymer polyethylenimine (PEI), a 40 kHz ultrasonic atomization frequency was used to formulate PLGA microparticles at a flow rate of 18 mL h1. High levels of gene expression and moderate cytotoxicity in COS-7 cells were achieved with the condensed pDNA at a nitrogen to phosphate (N/P) ratio of 20, thus demonstrating enhanced cellular uptake and expression of the transgene. The ability of the microparticles to convey pDNA was examined by characterizing the formulated microparticles. The microparticles displayed Z-average hydrodynamic diameters of 1.50-2.10 lm and zeta potentials of 17.8-23.2 mV. The encapsulation efficiencies were between 78 and 83%, and 76 and 85% of the embedded malaria pDNA molecules were released under physiological conditions in vitro. These results indicate that PLGA-mediated microparticles can be employed as potential gene delivery systems to antigen-presenting cells in the prevention of malaria.

Preparation and characterization of poly(lactic-co-glycolic acid) microparticles containing DNA molecules encoding a malaria vaccine candidate

Background A novel ultrasonic atomization approach for the formulation of biodegradable poly(lactic-co-glycolic acid) (PLGA) microparticles of a malaria DNA vaccine is presented. A 40 kHz ultrasonic atomization device was used to create the microparticles from a feedstock containing 5 volumes of 0.5% w/v PLGA in acetone and 1 volume of condensed DNA which was fed at a flow rate of 18ml h-1. The plasmid DNA vectors encoding a malaria protein were condensed with a cationic polymer before atomization. Results High levels of gene expression in vitro were observed in COS-7 cells transfected with condensed DNA at a nitrogen to phosphate (N/P) ratio of 10. At this N/P ratio, the condensed DNA exhibited a monodispersed nanoparticle size (Z-average diameter of 60.8 nm) and a highly positive zeta potential of 38.8mV. The microparticle formulations of malaria DNA vaccine were quality assessed and it was shown that themicroparticles displayed high encapsulation efficiencies between 82-96% and a narrow size distribution in the range of 0.8-1.9 μm. In vitro release profile revealed that approximately 82% of the DNA was released within 30 days via a predominantly diffusion controlledmass transfer system. Conclusions This ultrasonic atomization technique showed excellent particle size reproducibility and displayed potential as an industrially viable approach for the formulation of controlled release particles.

Creation of protein loaded biodegradable microparticles via ultrasonic atomization suitable for nasal delivery

Improved biopharmaceutical delivery may be achieved via the use of biodegradable microspheres as delivery vehicles. Biodegradable microspheres offer the advantages of maintaining sustained protein release over time whilst simultaneously protecting the biopharmaceutical from degradation. Particle samples produced by ultrasonic atomization were studied in order to determine a feed stock capable of producing protein loaded poly-ε-caprolactone (PCL) particles suitable for nasal delivery (i.e., less than 20 μm). A 40 kHz atomization system was used with a 6 mm full wave atomization probe. The effect of solids percent, feed flow rate, volumetric ratio of the polymer stock to the protein stock, and protein concentration in the protein stock on particle size characteristics were determined. It was shown that feed stocks containing 100 parts of 0.5 or 1.0% w/v PCL in acetone with one part 100 mg ml -1 BSA and 15 mg ml -1 PVA produced particles with a mass moment diameter (D[4,3]) of 13.17 μm and 9.10 μm, respectively in addition to displaying high protein encapsulation efficiencies of 93 and 95%, respectively. The biodegradable PCL particles were shown to be able to deliver encapsulated protein in vitro under physiological conditions.

Moving forwards with the aim of going backwards fast : high performance rowing as a learning environment

Background and Purpose: - This paper focuses on the learning culture within the high performance levels of rowing. In doing so, we explore the case of an individual’s learning as he moves across athletic, coaching and administrative functions. This exploration draws on a cultural learning framework and complementary theorisings related to reflexivity. Method - This study makes use of an intellectually, morally and collaboratively challenging approach whereby one member of the research team was also the sole participant of this study. The participant’s careers as a high performance athlete, coach and administrator, coupled with his experience in conducting empirical research presented a rare opportunity to engage in collaborative research (involving degrees of insider and outsider status for each of the research team). We acknowledge that others have looked to combine roles of coach / athlete / administrator with that of researcher however few (if any) have attempted to combine them all in one project. Moreover, coupled with the approach to reflexivity adopted in this study and the authorship contributions we consider this scholarly direction uncommon. Data were comprised of recorded research conversations, a subsequently constructed learning narrative, reflections on the narrative, a stimulated reflective piece from the participant, and a final (re)construction of the participant’s story. Accordingly, data were integrated through an iterative process of thematic analysis. Results - The cultural (i.e., the ways things get done) and structural (e.g., the rules and regulations) properties of high performance rowing were found to shape both the opportunities to be present (e.g., secure a place in the crew) and to learn (e.g., learn the skills required to perform at an Olympic level). However, the individual’s personal properties were brought to bear on re-shaping the constraints such that many limitations could be overcome. In keeping with the theory of learning cultures, the culture of rowing was found to position individuals (a coxswain in this case) differentially. In a similar manner, a range of structural features was found to be important in shaping the cultural and personal elements in performance contexts. For example, the ‘field of play’ was found to be important as a structural feature (i.e., inability of coach to communicate with athletes) in shaping the cultural and personal elements of learning in competition (e.g., positioning the coxswain as an in-boat coach and trusted crewmate). Finally, the cultural and structural elements in rowing appeared to be activated by the participant’s personal elements, most notably his orientation towards quality performance. Conclusion - The participant in this study was found to be driven by the project that he cares about most and at each turn he has bent his understanding of his sport back on itself to see if he can find opportunities to learn and subsequently explore ways to improve performance. The story here emphasises the importance of learner agency, and this is an aspect that has often been missing in recent theorising about learning. In this study, we find an agent using his ‘personal emergent powers to activate the resources in the culture and structure of his sport in an attempt to improve performance. We conclude from this account that this particular high performance rowing culture is one that provided support but nonetheless encouraged those involved, to ‘figure things out’ for themselves – be it as athletes, coaches and/or administrators.

High and low fear phenotypes show differences in phosphorylated (p44/42 ERK) MAPK-expressing neurons in the lateral amygdala

Post traumatic stress disorder (PTSD) is a serious medical condition effecting both military and civilian populations. While its etiology remains poorly understood it is characterized by high and prolonged levels of fear responding. One biological unknown is whether individuals expressing high or low conditioned fear memory encode the memory differently and if that difference underlies fear response. In this study we examined cellular mechanisms that underlie high and low conditioned fear behavior by using an advanced intercrossed mouse line (B6D2F1) selected for high and low Pavlovian fear response. A known requirement for consolidation of fear memory, phosphorylated mitogen activated protein kinase (p44/42 (ERK) MAPK (pMAPK)) in the lateral amygdala (LA) is a reliable marker of fear learning-related plasticity. In this study, we asked whether high and low conditioned fear behavior is associated with differential pMAPK expression in the LA and if so, is it due to an increase in neurons expressing pMAPK or increased pMAPK per neuron. To examine this, we quantified pMAPK-expressing neurons in the LA at baseline and following Pavlovian fear conditioning. Results indicate that high fear phenotype mice have more pMAPK-expressing neurons in the LA. This finding suggests that increased endogenous plasticity in the LA may be a component of higher conditioned fear responses and begins to explain at the cellular level how different fear responders encode fear memories. Understanding how high and low fear responders encode fear memory will help identify novel ways in which fear-related illness risk can be better predicted and treated.

Mice behaviorally selected for high Pavlovian fear memory exhibit fear generalization, HPA-axis alterations and increased basal activity in cortico-limbic circuits

Biological factors underlying individual variability in fearfulness and anxiety have important implications for stress-related psychiatric illness including PTSD and major depression. Using an advanced intercross line (AIL) derived from C57BL/6 and DBA/2J mouse strains and behavioral selection over 3 generations, we established two lines exhibiting High or Low fear behavior after fear conditioning. Across the selection generations, the two lines showed clear differences in training and tests for contextual and conditioned fear. Before fear conditioning training, there were no differences between lines in baseline freezing to a novel context. However, after fear conditioning High line mice demonstrated pronounced freezing in a new context suggestive of poor context discrimination. Fear generalization was not restricted to contextual fear. High fear mice froze to a novel acoustic stimulus while freezing in the Low line did not increase over baseline. Enhanced fear learning and generalization are consistent with transgenic and pharmacological disruption of the hypothalamic-pituitary-adrenal axis (HPA-axis) (Brinks, 2009, Thompson, 2004, Kaouane, 2012). To determine whether there were differences in HPA-axis regulation between the lines, morning urine samples were collected to measure basal corticosterone. Levels of secreted corticosterone in the circadian trough were analyzed by corticosterone ELISA. High fear mice were found to have higher basal corticosterone levels than low line animals. Examination of hormonal stress response components by qPCR revealed increased expression of CRH mRNA and decreased mRNA for MR and CRHR1 in hypothalamus of high fear mice. These alterations may contribute to both the behavioral phenotype and higher basal corticosterone in High fear mice. To determine basal brain activity in vivo in High and Low fear mice we used manganese-enhanced magnetic resonance imaging (MEMRI). Analysis revealed a pattern of basal brain activity made up of amygdala, cortical and hippocampal circuits that was elevated in the High line. Ongoing studies also seek to determine the relative balance of excitatory and inhibitory tone in the amygdala and hippocampus and the neuronal structure of its neurons. While these heterogeneous lines are selected on fear memory expression, HPA-axis alterations and differences in hippocampal activity segregate with the behavioral phenotypes. These differences are detectable in a basal state strongly suggesting these are biological traits underlying the behavioral phenotype (Johnson et al, 2011).