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.

A thermal expulsion approach to homogeneous large-volume methacrylate monolith preparation; enabling large-scale rapid purification of biomolecules

Numerous efforts have been dedicated to the synthesis of large-volume methacrylate monoliths for large-scale biomolecules purification but most were obstructed by the enormous release of exotherms during preparation, thereby introducing structural heterogeneity in the monolith pore system. A significant radial temperature gradient develops along the monolith thickness, reaching a terminal temperature that supersedes the maximum temperature required for structurally homogenous monoliths preparation. 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 polymerization in a gradual addition fashion. Characteristics of 80 mL monolith prepared using this technique was compared with that of a similar monolith synthesized in a bulk polymerization mode. An extra similarity in the radial temperature profiles was observed for the monolith synthesized via the heat expulsion technique. A maximum radial temperature gradient of only 4.3°C was recorded at the center and 2.1°C at the monolith peripheral for the combined heat expulsion and gradual addition technique. The comparable radial temperature distributions obtained birthed identical pore size distributions at different radial points along the monolith thickness.

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.

Cytokine secretion in macrophages : SNAREs, Rabs and membrane trafficking

Macrophages have the capacity to rapidly secrete a wide range of inflammatory mediators that influence the development and extent of an inflammatory response. Newly synthesized and/or preformed stored cytokines and other inflammatory mediators are released upon stimulation, the timing, and volume of which is highly regulated. To finely tune this process, secretion is regulated at many levels; at the level of transcription and translation and post-translationally at the endoplasmic reticulum (ER), Golgi, and at or near the cell surface. Here, we discuss recent advances in deciphering these cytokine pathways in macrophages, focusing on recent discoveries regarding the cellular machinery and mechanisms implicated in the synthesis, trafficking, and secretion of cytokines. The specific roles of trafficking machinery including chaperones, GTPases, cytoskeletal proteins, and SNARE membrane fusion proteins will be discussed.

Clinton, Bush, and Obama : changing policy and rhetoric in the United States Annual Trafficking in Persons Report

This chapter charts the political transitions in the anti-trafficking agenda and rhetoric of the U.S. Government across three Presidential administrations through a detailed examination of the annual Trafficking in Persons reports released by the Office to Monitor and Combat Trafficking in Persons between 2001 and 2012. We argue that the transitions in language and focus reflect key tensions that have dominated trafficking discourse throughout the Clinton, Bush and Obama Presidencies. These fissures include debate over law enforcement versus rights-based frameworks, competing approaches on victim protection and identification, and ongoing disputes about the relationship between prostitution and human trafficking.

Cytokine expression and secretion by skeletal muscle cells : regulatory mechanisms and exercise effects

Cytokines are important mediators of various aspects of health and disease, including appetite, glucose and lipid metabolism, insulin sensitivity, skeletal muscle hypertrophy and atrophy. Over the past decade or so, considerable attention has focused on the potential for regular exercise to counteract a range of disease states by modulating cytokine production. Exercise stimulates moderate to large increases in the circulating concentrations of interleukin (IL)-6, IL-8, IL-10, IL-1 receptor antagonist, granulocyte-colony stimulating factor, and smaller increases in tumor necrosis factor-α, monocyte chemotactic protein-1, IL-1β, brain-derived neurotrophic factor, IL-12p35/p40 and IL-15. Although many of these cytokines are also expressed in skeletal muscle, not all are released from skeletal muscle into the circulation during exercise. Conversely, some cytokines that are present in the circulation are not expressed in skeletal muscle after exercise. The reasons for these discrepant cytokine responses to exercise are unclear. In this review, we address these uncertainties by summarizing the capacity of skeletal muscle cells to produce cytokines, analyzing other potential cellular sources of circulating cytokines during exercise, and discussing the soluble factors and intracellular signaling pathways that regulate cytokine synthesis (e.g., RNA-binding proteins, microRNAs, suppressor of cytokine signaling proteins, soluble receptors).

Perspectives on Australian bankruptcy law through the prism of the World Bank report on the treatment of the insolvency of natural persons

The insolvency of natural persons raises questions not only for a nation’s economy but also for its concern for equity. The World Bank has recently released a Report on the Treatment of the Insolvency of Natural Persons to guide nations in addressing the issues raised by an individual debtor’s insolvency. A brief review of Australia’s personal insolvency laws shows that it addresses many of the issues raised by the Report. However two areas are identified as worthy of further investigation by policy-makers and scholars to better address a concern for equity.

The competitive market for assurance engagements on greenhouse gas statements : is there a role for assurers from the accounting profession?

In response to current and increasing demand for assurance on greenhouse gas statements, the International Auditing and Assurance Standards Board (IAASB) released an exposure draft of a new assurance standard, ISAE 3410 'Assurance on a Greenhouse Gas Statement' (IFAC 2011), to provide comprehensive guidance on these types of greenhouse gas (GHG) assurance engagements. Internationally, approximately 50 percent of GHG statements are independently assured. The related assurance market is competitive, with the accounting profession and those outside the profession currently holding approximately equal shares. This paper highlights the characteristics of GHG assurance engagements that warrant multi-disciplinary teamwork, the unique and interdependent skill-sets that different practitioners bring to these engagements, and the market forces that create a demand for diverse providers.

TriggerSync: A time synchronisation tool

This paper presents a framework for synchronising multiple triggered sensors with respect to a local clock using standard computing hardware. Providing sensor measurements with accurate and meaningful timestamps is important for many sensor fusion, state estimation and control applications. Accurately synchronising sensor timestamps can be performed with specialised hardware, however, performing sensor synchronisation using standard computing hardware and non-real-time operating systems is difficult due to inaccurate and temperature sensitive clocks, variable communication delays and operating system scheduling delays. Results show the ability of our framework to estimate time offsets to sub-millisecond accuracy. We also demonstrate how synchronising timestamps with our framework results in a tenfold reduction in image stabilisation error for a vehicle driving on rough terrain. The source code will be released as an open source tool for time synchronisation in ROS.

Love hurts: The costly reality of online romance fraud

Online dating and romance scams continue to lure in Australians with figures this week showing people have lost more than A$23 million this year alone, with average individual losses at A$21,000 – three times higher than other types of fraud. The Australian Competition and Consumer Commission (ACCC) set up the Scam Disruption Project in August to help target those it believes have been caught in such scams. Over three months it sent 1,500 letters to potential victims in New South Wales and the Australian Capital Territory. The figures released this week show that 50 people have been scammed, losing a total A$1.7 million – that’s an average of A$34,000 per victim. Almost three quarters of the scams were dating and romance related, which saw it evolve into the number one category of fraud victimisation. Romance scams continue to pose a problem – despite the efforts of the police and ACCC – so why is it that people continue to fall for them?

A ‘Big Data’ Approach to Mapping the Australian Twittersphere

This chapter discusses the methodological aspects and empirical findings of a large-scale, funded project investigating public communication through social media in Australia. The project concentrates on Twitter, but we approach it as representative of broader current trends toward the integration of large datasets and computational methods into media and communication studies in general, and social media scholarship in particular. The research discussed in this chapter aims to empirically describe networks of affiliation and interest in the Australian Twittersphere, while reflecting on the methodological implications and imperatives of ‘big data’ in the humanities. Using custom network crawling technology, we have conducted a snowball crawl of Twitter accounts operated by Australian users to identify more than one million users and their follower/followee relationships, and have mapped their interconnections. In itself, the map provides an overview of the major clusters of densely interlinked users, largely centred on shared topics of interest (from politics through arts to sport) and/or sociodemographic factors (geographic origins, age groups). Our map of the Twittersphere is the first of its kind for the Australian part of the global Twitter network, and also provides a first independent and scholarly estimation of the size of the total Australian Twitter population. In combination with our investigation of participation patterns in specific thematic hashtags, the map also enables us to examine which areas of the underlying follower/followee network are activated in the discussion of specific current topics – allowing new insights into the extent to which particular topics and issues are of interest to specialised niches or to the Australian public more broadly. Specifically, we examine the Twittersphere footprint of dedicated political discussion, under the #auspol hashtag, and compare it with the heightened, broader interest in Australian politics during election campaigns, using #ausvotes; we explore the different patterns of Twitter activity across the map for major television events (the popular competitive cooking show #masterchef, the British #royalwedding, and the annual #stateoforigin Rugby League sporting contest); and we investigate the circulation of links to the articles published by a number of major Australian news organisations across the network. Such analysis, which combines the ‘big data’-informed map and a close reading of individual communicative phenomena, makes it possible to trace the dynamic formation and dissolution of issue publics against the backdrop of longer-term network connections, and the circulation of information across these follower/followee links. Such research sheds light on the communicative dynamics of Twitter as a space for mediated social interaction. Our work demonstrates the possibilities inherent in the current ‘computational turn’ (Berry, 2010) in the digital humanities, as well as adding to the development and critical examination of methodologies for dealing with ‘big data’ (boyd and Crawford, 2011). Out tools and methods for doing Twitter research, released under Creative Commons licences through our project Website, provide the basis for replicable and verifiable digital humanities research on the processes of public communication which take place through this important new social network.

Keeping kids in school is not as simple as carrots and sticks

The OECD’s 2009 Jobs for Youth report, released on the heels of the global financial crisis, made a number of policy recommendations to the Australian government to prevent a rise in youth unemployment. Five years on, youth unemployment has risen from around 8% to 14.8%, and around one in five children do not complete year 12, according to data from the National Centre for Vocational Education Research...

Evolved gas analysis of coal-derived pyrite/marcasite

The products evolved during the thermal decomposition of the coal-derived pyrite/marcasite were studied using simultaneous thermogravimetry coupled with Fourier-transform infrared spectroscopy and mass spectrometry (TG-FTIR–MS) technique. The main gases and volatile products released during the thermal decomposition of the coal-derived pyrite/marcasite are water (H2O), carbon dioxide (CO2), and sulfur dioxide (SO2). The results showed that the evolved products obtained were mainly divided into two processes: (1) the main evolved product H2O is mainly released at below 300 °C; (2) under the temperature of 450–650 °C, the main evolved products are SO2 and small amount of CO2. It is worth mentioning that SO3 was not observed as a product as no peak was observed in the m/z = 80 curve. The chemical substance SO2 is present as the main gaseous product in the thermal decomposition for the sample. The coal-derived pyrite/marcasite is different from mineral pyrite in thermal decomposition temperature. The mass spectrometric analysis results are in good agreement with the infrared spectroscopic analysis of the evolved gases. These results give the evidence on the thermal decomposition products and make all explanations have the sufficient evidence. Therefore, TG–MS–IR is a powerful tool for the investigation of gas evolution from the thermal decomposition of materials.

Exploring with Grammar in the Primary Years: Learning about Language in the Australian Curriculum: English

This new volume, Exploring with Grammar in the Primary Years (Exley, Kevin & Mantei, 2014), follows on from Playing with Grammar in the Early Years (Exley & Kervin, 2013). We extend our thanks to the ALEA membership for their take up of the first volume and the vibrant conversations around our first attempt at developing a pedagogy for the teaching of grammar in the early years. Your engagement at locally held ALEA events has motivated us to complete this second volume and reassert our interest in the pursuit of socially-just outcomes in the primary years. As noted in Exley and Kervin (2013), we believe that mastering a range of literacy competences includes not only the technical skills for learning, but also the resources for viewing and constructing the world (Freire and Macdeo, 1987). Rather than seeing knowledge about language as the accumulation of technical skills alone, the viewpoint to which we subscribe treats knowledge about language as a dialectic that evolves from, is situated in, and contributes to active participation within a social arena (Halliday, 1978). We acknowledge that to explore is to engage in processes of discovery as we look closely and examine the opportunities before us. As such, we draw on Janks’ (2000; 2014) critical literacy theory to underpin many of the learning experiences in this text. Janks (2000) argues that effective participation in society requires knowledge about how the power of language promotes views, beliefs and values of certain groups to the exclusion of others. Powerful language users can identify not only how readers are positioned by these views, but also the ways these views are conveyed through the design of the text, that is, the combination of vocabulary, syntax, image, movement and sound. Similarly, powerful designers of texts can make careful modal choices in written and visual design to promote certain perspectives that position readers and viewers in new ways to consider more diverse points of view. As the title of our text suggests, our activities are designed to support learners in exploring the design of texts to achieve certain purposes and to consider the potential for the sharing of their own views through text production. In Exploring with Grammar in the Primary Years, we focus on the Year 3 to Year 6 grouping in line with the Australian Curriculum, Assessment and Reporting Authority’s (hereafter ACARA) advice on the ‘nature of learners’ (ACARA, 2014). Our goal in this publication is to provide a range of highly practical strategies for scaffolding students’ learning through some of the Content Descriptions from the Australian Curriculum: English Version 7.2, hereafter AC:E (ACARA, 2014). We continue to express our belief in the power of using whole texts from a range of authentic sources including high quality children’s literature, the internet, and examples of community-based texts to expose students to the richness of language. Taking time to look at language patterns within actual texts is a pathway to ‘…capture interest, stir the imagination and absorb the [child]’ into the world of language and literacy (Saxby, 1993, p. 55). It is our intention to be more overt this time and send a stronger message that our learning experiences are simply ‘sample’ activities rather than a teachers’ workbook or a program of study to be followed. We’re hoping that teachers and students will continue to explore their bookshelves, the internet and their community for texts that provide powerful opportunities to engage with language-based learning experiences. In the following three sections, we have tried to remain faithful to our interpretation of the AC:E Content Descriptions without giving an exhaustive explanation of the grammatical terms. This recently released curriculum offers a new theoretical approach to building students’ knowledge about language. The AC:E uses selected traditional terms through an approach developed in systemic functional linguistics (see Halliday and Matthiessen, 2004) to highlight the dynamic forms and functions of multimodal language in texts. For example, the following statement, taken from the ‘Language: Knowing about the English language’ strand states: English uses standard grammatical terminology within a contextual framework, in which language choices are seen to vary according to the topics at hand, the nature and proximity of the relationships between the language users, and the modalities or channels of communication available (ACARA, 2014). Put simply, traditional grammar terms are used within a functional framework made up of field, tenor, and mode. An understanding of genre is noted with the reference to a ‘contextual framework’. The ‘topics at hand’ concern the field or subject matter of the text. The ‘relationships between the language users’ is a description of tenor. There is reference to ‘modalities’, such as spoken, written or visual text. We posit that this innovative approach is necessary for working with contemporary multimodal and cross-cultural texts (see Exley & Mills, 2012). Other excellent tomes, such as Derewianka (2011), Humphrey, Droga and Feez (2012), and Rossbridge and Rushton (2011) provide more comprehensive explanations of this unique metalanguage, as does the AC:E Glossary. We’ve reproduced some of the AC:E Glossary at the end of this publication. We’ve also kept the same layout for our learning experiences, ensuring that our teacher notes are not only succinct but also prudent in their placement. Each learning experience is connected to a Content Description from the AC:E and contains an experience with an identified purpose, suggested resource text and a possible sequence for the experience that always commences with an orientation to text followed by an examination of a particular grammatical resource. Our plans allow for focused discussion, shared exploration and opportunities to revisit the same text for the purpose of enhancing meaning making. Some learning experiences finish with deconstruction of a stimulus text while others invite students to engage in the design of new texts. We encourage you to look for opportunities in your own classrooms to move from text deconstruction to text design. In this way, students can express not only their emerging grammatical understandings, but also the ways they might position readers or viewers through the creation of their own texts. We expect that each of these learning experiences will vary in the time taken. Some may indeed take a couple if not a few teaching episodes to work through, especially if students are meeting a concept or a pedagogical strategy for the first time. We hope you use as much, or as little, of each experience as is needed for your students. We do not want the teaching of grammar to slip into a crisis of irrelevance or to be seen as a series of worksheet drills with finite answers. We firmly believe that strategies for effective deconstruction and design practice, however, have much portability. We three are very keen to hear from teachers who are adopting and adapting these learning experiences in their classrooms. Please email us on b.exley@qut.edu.au, lkervin@uow.edu.au or jessicam@ouw.edu.au. We’d love to continue the conversation with you over time. Beryl Exley, Lisa Kervin & Jessica Mantei