The co-transcriptome of uropathogenic Escherichia coli-infected mouse macrophages reveals new insights into host-pathogen interactions

Urinary tract infections (UTI) are among the most common infections in humans. Uropathogenic Escherichia coli (UPEC) can invade and replicate within bladder epithelial cells, and some UPEC strains can also survive within macrophages. To understand the UPEC transcriptional program associated with intramacrophage survival, we performed host–pathogen co-transcriptome analyses using RNA sequencing. Mouse bone marrow-derived macrophages (BMMs) were challenged over a 24 h time course with two UPEC reference strains that possess contrasting intramacrophage phenotypes: UTI89, which survives in BMMs, and 83972, which is killed by BMMs. Neither of these strains caused significant BMM cell death at the low multiplicity of infection that was used in this study. We developed an effective computational framework that simultaneously separated, annotated, and quantified the mammalian and bacterial transcriptomes. BMMs responded to the two UPEC strains with a broadly similar gene expression program. In contrast, the transcriptional responses of the UPEC strains diverged markedly from each other. We identified UTI89 genes upregulated at 24 h post-infection, and hypothesized that some may contribute to intramacrophage survival. Indeed, we showed that deletion of one such gene (pspA) significantly reduced UTI89 survival within BMMs. Our study provides a technological framework for simultaneously capturing global changes at the transcriptional level in co-cultures, and has generated new insights into the mechanisms that UPEC use to persist within the intramacrophage environment.

Influences of molecular structure on co-crystallization of polypyridyl metal complexes.

Heteroleptic complexes of the type \[RuL2L′](PF6)2 (L, L′ = combinations of 1,10-phenanthroline (phen) and 2,2′-bipyridine (bipy)) were found to cocrystallize with \[Ni(phen)3](PF6)2 to produce cocrystals of \[Ni(phen)3]x\[RuL2L′]1–x(PF6)2. In this report we show that the ability of the complexes to cocrystallize is influenced by the number of common ligands between complexes in solution. Supramolecular selection is a phenomenon caused by molecular recognition through which cocrystals can grow from the same solution but contain different ratios of the molecular components. It was found that systems where L = phen displayed less supramolecular selection than systems where L = bipy. With increasing supramolecular selection, the composition of cocrystals was found to vary significantly from the initial relative concentration in the cocrystallizing solution, and therefore it was increasingly difficult to control the final composition of the resultant cocrystals. Consequently, modulation of concentration-dependent properties such as phase was also found to be less predictable with increasing supramolecular selection. Notwithstanding the complication afforded by the presence of supramolecular selection, our results reaffirm the robustness of the \[M(phen)3](PF6)2 structure because it was maintained even when ca. 90% of the complexes in the cocrystals were \[Ru(phen)(bipy)2](PF6)2, which in its pure form is not isomorphous with \[M(phen)3](PF6)2. Experiments between complexes without common ligands, i.e., \[Ru(bipy)3](PF6)2 cocrystallized with \[Ni(phen)3](PF6)2, were found to approach the limit to which molecular recognition processes can be confused into cocrystallizing different molecules to form single cocrystals. For these systems the result was the formation of block-shaped crystals skewered by a needle-shaped crystals.

Understanding preservice teachers’ development of pedagogical knowledge practices when co-teaching primary science to peers

Preservice teachers articulate the need for more teaching experiences for developing their practices, however, extending beyond existing school arrangements may present difficulties. Thus, it is important to understand preservice teachers’ development of pedagogical knowledge practices when in the university setting. This mixed-method study investigated 48 second-year preservice teachers’ development of pedagogical knowledge practices as a result of co-teaching primary science to peers. Data were collected through a survey, video-recorded lessons, extended written responses and researcher observations. The study showed how these preservice teachers demonstrated 9 of 11 pedagogical knowledge practices within the co-teaching arrangement. However, research is needed to determine the level of development on each pedagogical knowledge practice and how these practices can be transferred into authentic primary classroom settings.

Opportunities and challenges in engaging citizens in the co-production of infrastructure-based public services in Australia

Research and practice have observed a shift towards service-oriented approaches that depend on input from citizens as co-producers of services. Yet in the delivery of public infrastructure the focus is still on managing assets rather than services. Using a Policy Delphi approach, we found that although experts advocate service-centric approaches guidelines and policies lack a service-centric perspective. Findings revealed a range of impediments to effective stakeholder involvement. The paper contributes to co-production and new public governance literature and offers directions for public infrastructure decision-makers to support and reconnect disengaged government–citizen relations, and determine ways of understanding optimal service outcomes.

Synthesis of biodegradable polymer-mesoporous silica composite microspheres for DNA prime-protein boost vaccination

DNA vaccines or proteins are capable of inducing specific immunity; however, the translation to the clinic has generally been problematic, primarily due to the reduced magnitude of immune response and poor pharmacokinetics. Herein we demonstrate a composite microsphere formulation, composed of mesoporous silica spheres (MPS) and poly(d,l-lactide-co-glycolide) (PLGA), enables the controlled delivery of a prime-boost vaccine via the encapsulation of plasmid DNA (pDNA) and protein in different compartments. Method with modified dual-concentric-feeding needles attached to a 40 kHz ultrasonic atomizer was studied. These needles focus the flow of two different solutions, which passed through the ultrasonic atomizer. The process synthesis parameters, which are important to the scale-up of composite microspheres, were also studied. These parameters include polymer concentration, feed flowrate, and volumetric ratio of polymer and pDNA-PEI/MPS-BSA. This fabrication technique produced composite microspheres with mean D[4,3] ranging from 6 to 34 μm, depending upon the microsphere preparation. The resultant physical morphology of composite microspheres was largely influenced by the volumetric ratio of pDNA-PEI/MPS-BSA to polymer, and this was due to the precipitation of MPS at the surface of the microspheres. The encapsulation efficiencies were predominantly in the range of 93-98% for pDNA and 46-68% for MPS. In the in vitro studies, the pDNA and protein showed different release kinetics in a 40 day time frame. The dual-concentric-feeding in ultrasonic atomization was shown to have excellent reproducibility. It was concluded that this fabrication technique is an effective method to prepare formulations containing a heterologous prime-boost vaccine in a single delivery system.

Versatility of polymethacrylate monoliths for chromatographic purification of biomolecules

Polymethacrylate monoliths, specifically poly(glycidyl methacrylate-co-ethylene dimethacrylate) or poly(GMA-co-EDMA) monoliths, are a new generation of chromatographic supports and are significantly different from conventional particle-based adsorbents, membranes, and other monolithic supports for biomolecule purification. Similar to other monoliths, polymethacrylate monoliths possess large pores which allow convective flow of mobile phase and result in high flow rates at reduced pressure drop, unlike particulate supports. The simplicity of the adsorbent synthesis, pH resistance, and the ease and flexibility of tailoring their pore size to that of the target biomolecule are the key properties which differentiate polymethacrylate monoliths from other monoliths. Polymethacrylate monoliths are endowed with reactive epoxy groups for easy functionalization (with anion-exchange, hydrophobic, and affinity ligands) and high ligand retention. In this review, the structure and performance of polymethacrylate monoliths for chromatographic purification of biomolecules are evaluated and compared to those of other supports. The development and use of polymethacrylate monoliths for research applications have grown rapidly in recent times and have enabled the achievement of high through-put biomolecule purification on semi-preparative and preparative scales.

Process considerations related to the microencapsulation of plasmid DNA via ultrasonic atomization

An effective means of facilitating DNA vaccine delivery to antigen presenting cells is through biodegradable microspheres. Microspheres offer distinct advantages over other delivery technologies by providing release of DNA vaccine in its bioactive form in a controlled fashion. In this study, biodegradable poly(D,L-lactide-coglycolide) (PLGA) microspheres containing polyethylenimine (PEI) condensed plasmid DNA (pDNA) were prepared using a 40 kHz ultrasonic atomization system. Process synthesis parameters, which are important to the scale-up of microspheres that are suitable for nasal delivery (i.e., less than 20 μm), were studied. These parameters include polymer concentration; feed flowrate; volumetric ratio of polymer and pDNA-PEI (plasmid DNA-polyethylenimine) complexes; and nitrogen to phosphorous (N/P) ratio. PDNA encapsulation efficiencies were predominantly in the range 82-96%, and the mean sizes of the particle were between 6 and 15 μm. The ultrasonic synthesis method was shown to have excellent reproducibility. PEI affected morphology of the microspheres, as it induced the formation of porous particles that accelerate the release rate of pDNA. The PLGA microspheres displayed an in vitro release of pDNA of 95-99% within 30 days and demonstrated zero order release kinetics without an initial spike of pDNA. Agarose electrophoresis confirmed conservation of the supercoiled form of pDNA throughout the synthesis and in vitro release stages. It was concluded that ultrasonic atomization is an efficient technique to overcome the key obstacles in scaling-up the manufacture of encapsulated vaccine for clinical trials and ultimately, commercial applications.

Storing energy in plastics : a review on conducting polymers & their role in electrochemical energy storage

Conducting polymers have become the focus of research due to their interesting properties, such as a wide range of conductivity, facile production, mechanical stability, light weight and low cost and due to the ease with which conducting polymers can be nanostructured to meet the specific application. They have become valuable materials for many applications, such as energy storage and generation. Recently, conducting polymers have been studied to be used in supercapacitors, battery electrode and fuel cells. This article is to briefly discuss the background & theory behind their conductivity as well as to highlight the recent contributions of conducting polymers to the field of energy and their significance. Furthermore, the methods of production of the conducting polymers in addition to the different ways utilised to nano-engineer special morphologies are discussed.

Co-optimisation of indoor environmental quality and energy consumption within urban office buildings

This study aimed to develop a multi-component model that can be used to maximise indoor environmental quality inside mechanically ventilated office buildings, while minimising energy usage. The integrated model, which was developed and validated from fieldwork data, was employed to assess the potential improvement of indoor air quality and energy saving under different ventilation conditions in typical air-conditioned office buildings in the subtropical city of Brisbane, Australia. When operating the ventilation system under predicted optimal conditions of indoor environmental quality and energy conservation and using outdoor air filtration, average indoor particle number (PN) concentration decreased by as much as 77%, while indoor CO2 concentration and energy consumption were not significantly different compared to the normal summer time operating conditions. Benefits of operating the system with this algorithm were most pronounced during the Brisbane’s mild winter. In terms of indoor air quality, average indoor PN and CO2 concentrations decreased by 48% and 24%, respectively, while potential energy savings due to free cooling went as high as 108% of the normal winter time operating conditions. The application of such a model to the operation of ventilation systems can help to significantly improve indoor air quality and energy conservation in air-conditioned office buildings.

Game On: Know Alcohol - Co-creating a tailored alcohol social marketing program

Research demonstrates that education programs are more effective when their target audiences and other key stakeholder groups are involved in the design. The majority of programs continue to be researcher and expert driven. This study extends upon previous research by employing a co-creation research design to offer a consumer driven alternative to education program design. Two co-creation groups involving twenty 14-16 year old Year 10 students who had previously participated in the Game On:Know Alcohol (GOKA) program, which aims to moderate alcohol drinking attitudes and behaviour, were conducted. Analysis revealed that a co-created GOKA program will differ substantially from the researcher and expert driven program that is currently being field tested. Students prefer interactive activities and activities that engage and challenge. Co-creation offers the potential to contest researcher and expert views and may offer to assist in the generation of new insights for the development of education programs.

The Business of Unfinished Business : Reflections on Co-construction of Meanings in Research Encounters

My concern in this commentary is the discrepancy between cultural psychologists' theoretical claims that meanings are co-constructed by, with and for individuals in ongoing social interaction, and their research practices where researcher's and research participant's meaning-making processes are separated in time into sequential turns. I argue for the need to live up to these theoretical assumptions, by making both the initial research encounter and the researcher's later interpretation process more co-constructive. I suggest making the initial research encounter more co-constructive by paying attention to these moments when the negotiated flow of interaction between researcher and research participant breaks down, for it allows the research participant's meaning-making to be traced and makes the researcher's efforts towards meaning more explicit. I propose to make the later interpretation process more co-constructive by adopting a more open-ended and dialogical way of writing that is specifically addressed to research participants and invites them to actively engage with researcher's meaning-making.