Comparison of cervical and blood T-cell responses to human papillomavirus-16 in women with human papillomavirus-associated cervical intraepithelial neoplasia

Human papillomaviruses (HPVs) are obligate epithelial pathogens and typically cause localized mucosal infections. We therefore hypothesized that T-cell responses to HPV antigens would be greater at sites of pathology than in the blood. Focusing on HPV-16 because of its association with cervical cancer, the magnitude of HPV-specific T-cell responses at the cervix was compared with those in the peripheral blood by intracellular cytokine staining following direct ex vivo stimulation with both virus-like particles assembled from the major capsid protein L1, and the major HPV oncoprotein, E7. We show that both CD4 + and CD8 + T cells from the cervix responded to the HPV-16 antigens and that interferon-γ (IFN-γ) production was HPV type-specific. Comparing HPV-specific T-cell IFN-γ responses at the cervix with those in the blood, we found that while CD4 + and CD8 + T-cell responses to L1 were significantly correlated between compartments (P = 0.02 and P = 0.05, respectively), IFN-γ responses in both T-cell subsets were significantly greater in magnitude at the cervix than in peripheral blood (P = 0.02 and P = 0.003, respectively). In contrast, both CD4 + and CD8 + T-cell IFN-γ responses to E7 were of similar magnitude in both compartments and CD8 + responses were significantly correlated between these distinct immunological compartments (P = 0.04). We therefore show that inflammatory T-cell responses against L1 (but not E7) demonstrate clear compartmental bias and the magnitude of these responses do reflect local viral replication but that correlation of HPV-specific responses between compartments indicates their linkage.

Optimization of chimeric HIV-1 virus-like particle production in a baculovirus-insect cell expression system

A baculovirus-insect cell expression system potentially provides the means to produce prophylactic HIV-1 virus-like particle (VLP) vaccines inexpensively and in large quantities. However, the system must be optimized to maximize yields and increase process efficiency. In this study, we optimized the production of two novel, chimeric HIV-1 VLP vaccine candidates (GagRT and GagTN) in insect cells. This was done by monitoring the effects of four specific factors on VLP expression: these were insect cell line, cell density, multiplicity of infection (MOI), and infection time. The use of western blots, Gag p24 ELISA, and four-factorial ANOVA allowed the determination of the most favorable conditions for chimeric VLP production, as well as which factors affected VLP expression most significantly. Both VLP vaccine candidates favored similar optimal conditions, demonstrating higher yields of VLPs when produced in the Trichoplusia ni Pro insect cell line, at a cell density of 1 × 106 cells/mL, and an infection time of 96 h post infection. It was found that cell density and infection time were major influencing factors, but that MOI did not affect VLP expression significantly. This work provides a potentially valuable guideline for HIV-1 protein vaccine optimization, as well as for general optimization of a baculovirus-based expression system to produce complex recombinant proteins. © 2009 American Institute of Chemical Engineers.

Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

Background Insect baculovirus-produced Human immunodeficiency virus type 1 (HIV-1) Gag virus-like-particles (VLPs) stimulate good humoral and cell-mediated immune responses in animals and are thought to be suitable as a vaccine candidate. Drawbacks to this production system include contamination of VLP preparations with baculovirus and the necessity for routine maintenance of infectious baculovirus stock. We used piggyBac transposition as a novel method to create transgenic insect cell lines for continuous VLP production as an alternative to the baculovirus system. Results Transgenic cell lines maintained stable gag transgene integration and expression up to 100 cell passages, and although the level of VLPs produced was low compared to baculovirus-produced VLPs, they appeared similar in size and morphology to baculovirus-expressed VLPs. In a murine immunogenicity study, whereas baculovirus-produced VLPs elicited good CD4 immune responses in mice when used to boost a prime with a DNA vaccine, no boost response was elicited by transgenically produced VLPs. Conclusion Transgenic insect cells are stable and can produce HIV Pr55 Gag VLPs for over 100 passages: this novel result may simplify strategies aimed at making protein subunit vaccines for HIV. Immunogenicity of the Gag VLPs in mice was less than that of baculovirus-produced VLPs, which may be due to lack of baculovirus glycoprotein incorporation in the transgenic cell VLPs. Improved yield and immunogenicity of transgenic cell-produced VLPs may be achieved with the addition of further genetic elements into the piggyBac integron.

Ruptures in the production of girlhood : teenage flows and multiplicities [Abstract]

This paper examines the paradoxical and ubiquitous nature of Butler’s heterosexual matrix, and opens it up to an alternative Deleuzian analysis. Drawing on stories and art works produced in a collective biography workshop on girls and sexuality this paper extends previous work on the subversion of the heterosexual matrix undertaken by Renold and Ringrose (2008). The paper moves, as they do, from a molar to a molecular analysis, but extends that work by re-thinking the girl/subject in terms of Deleuze and Guattari’s endlessly transforming multiplicities where “the self is only a threshold, a door, a becoming between two multiplicities” (Deleuze and Guattari, 1987: 249)

Ionic liquid pre-treatment and fractionation of sugarcane bagasse for the production of bioethanol

Pretretament is an essential and expensive processing step for the manufacturing of ethanol from lignocellulosic raw materials. Ionic liquids are a new class of solvents that have the potential to be used as pretreatment agents. The attractive characteristics of ionic liquid pretreatment of lignocellulosics such as thermal stability, dissolution properties, fractionation potential, cellulose decrystallisation capacity and saccharification impact are investigated in this thesis. Dissolution of bagasse with 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) at high temperatures (110 �‹C to 160 �‹C) is investigated as a pretreatment process. Material balances are reported and used along with enzymatic saccharification data to identify optimum pretreatment conditions (150 �‹C for 90 min). At these conditions, the dissolved and reprecipitated material is enriched in cellulose, has a low crystallinity and the cellulose component is efficiently hydrolysed (93 %, 3 h, 15 FPU). At pretreatment temperatures < 150 �‹C, the undissolved material has only slightly lower crystallinity than the starting. At pretreatment temperatures . 150 �‹C, the undissolved material has low crystallinity and when combined with the dissolved material has a saccharification rate and extent similar to completely dissolved material (100 %, 3h, 15 FPU). Complete dissolution is not necessary to maximize saccharification efficiency at temperatures . 150 �‹C. Fermentation of [C4mim]Cl-pretreated, enzyme-saccharified bagasse to ethanol is successfully conducted (85 % molar glucose-to-ethanol conversion efficiency). As compared to standard dilute acid pretreatment, the optimised [C4mim]Cl pretreatment achieves substantially higher ethanol yields (79 % cf. 52 %) in less than half the processing time (pretreatment, saccharification, fermentation). Fractionation of bagasse partially dissolved in [C4mim]Cl to a polysaccharide rich and a lignin rich fraction is attempted using aqueous biphasic systems (ABSs) and single phase systems with preferential precipitation. ABSs of ILs and concentrated aqueous inorganic salt solutions are achievable (e.g. [C4mim]Cl with 200 g L-1 NaOH), albeit they exhibit a number of technical problems including phase convergence (which increases with increasing biomass loading) and deprotonation of imidazolium ILs (5 % - 8 % mol). Single phase fractionation systems comprising lignin solvents / cellulose antisolvents, viz. NaOH (2M) and acetone in water (1:1, volume basis), afford solids with, respectively, 40 % mass and 29 % mass less lignin than water precipitated solids. However, this delignification imparts little increase in saccharification rates and extents of these solids. An alternative single phase fractionation system is achieved simply by using water as an antisolvent. Regulating the water : IL ratio results in a solution that precipitates cellulose and maintains lignin in solution (0.5 water : IL mass ratio) in both [C4mim]Cl and 1-ethyl-3-methylimidazolium acetate ([C2mim]OAc)). This water based fractionation is applied in three IL pretreatments on bagasse ([C4mim]Cl, 1-ethyl-3-methyl imidazolium chloride ([C2mim]Cl) and [C2mim]OAc). Lignin removal of 10 %, 50 % and 60 % mass respectively is achieved although only 0.3 %, 1.5 % and 11.7 % is recoverable even after ample water addition (3.5 water : IL mass ratio) and acidification (pH . 1). In addition the recovered lignin fraction contains 70 % mass hemicelluloses. The delignified, cellulose-rich bagasse recovered from these three ILs is exposed to enzyme saccharification. The saccharification (24 h, 15 FPU) of the cellulose mass in starting bagasse, achieved by these pretreatments rank as: [C2mim]OAc (83 %)>>[C2mim]Cl (53 %)=[C4mim]Cl(53%). Mass balance determinations accounted for 97 % of starting bagasse mass for the [C4mim]Cl pretreatment , 81 % for [C2mim]Cl and 79 %for [C2mim]OAc. For all three IL treatments, the remaining bagasse mass (not accounted for by mass balance determinations) is mainly (more than half) lignin that is not recoverable from the liquid fraction. After pretreatment, 100 % mass of both ions of all three ILs were recovered in the liquid fraction. Compositional characteristics of [C2mim]OAc treated solids such as low lignin, low acetyl group content and preservation of arabinosyl groups are opposite to those of chloride IL treated solids. The former biomass characteristics resemble those imparted by aqueous alkali pretreatment while the latter resemble those of aqueous acid pretreatments. The 100 % mass recovery of cellulose in [C2mim]OAc as opposed to 53 % mass recovery in [C2mim]Cl further demonstrates this since the cellulose glycosidic bonds are protected under alkali conditions. The alkyl chain length decrease in the imidazolium cation of these ILs imparts higher rates of dissolution and losses, and increases the severity of the treatment without changing the chemistry involved.

The Australian screen producer: Understanding screen production cultures in a period of industrial transition [Abstract]

The screen producer plays a vital role in shaping the creative, commercial and entrepreneurial dimensions of production. And yet Australian film history is most often presented as an appreciation of film directors or an examination of industrial governance measures. On the other hand, public funding agencies in Australia have, for the most part, supported independent film and television production as a producer-led, or producer-as-auteur production system, and as such the producer has played a critical role in shaping the broader independent production landscape. In recent years, fundamental changes to distribution and consumption practices have had a major impact on the nature of screen production. Screen producers are increasingly migrating into emerging online, transmedia and cross-media production; generating both opportunities and challenges for traditional producers. However, the production cultures and motivations of producers operating in these emergent spaces remain poorly understood. This presentation will focus on the largely unremarked role of the producer in Australian screen scholarship. It will explore the ways in which the practice of screen producing is evolving and the migratory pathways of traditional producers moving into digital/new media production. The presentation’s primary findings are drawn from the 2011 Australian Screen Producer Survey; a national study of the activities of Australian screen producers conducted by the ARC Centre of Excellence for Creative Industries and Innovation (CCI), Queensland University of Technology, with support from the Centre for Screen Business /Australian Film Television and Radio School (AFTRS). From longitudinal analysis, the presentation will compare and contrast data from the 2009 and 2011 survey across film, television, corporate production and new media industry segments. In so doing the presentation will delineate the practices, attitudes, strategies, and aspirations of screen producers operating in a convergent digital media marketplace and suggest ways forward for a more industrially cognisant approach to screen history.

Bioreactor for blood product production

The feasibility of ex vivo blood production is limited by both biological and engineering challenges. From an engineering perspective, these challenges include the significant volumes required to generate even a single unit of a blood product, as well as the correspondingly high protein consumption required for such large volume cultures. Membrane bioreactors, such as hollow fiber bioreactors (HFBRs), enable cell densities approximately 100-fold greater than traditional culture systems and therefore may enable a significant reduction in culture working volumes. As cultured cells, and larger molecules, are retained within a fraction of the system volume, via a semipermeable membrane it may be possible to reduce protein consumption by limiting supplementation to only this fraction. Typically, HFBRs are complex perfusion systems having total volumes incompatible with bench scale screening and optimization of stem cell-based cultures. In this article we describe the use of a simplified HFBR system to assess the feasibility of this technology to produce blood products from umbilical cord blood-derived CD34+ hematopoietic stem progenitor cells (HSPCs). Unlike conventional HFBR systems used for protein manufacture, where cells are cultured in the extracapillary space, we have cultured cells in the intracapillary space, which is likely more compatible with the large-scale production of blood cell suspension cultures. Using this platform we direct HSPCs down the myeloid lineage, while targeting a 100-fold increase in cell density and the use of protein-free bulk medium. Our results demonstrate the potential of this system to deliver high cell densities, even in the absence of protein supplementation of the bulk medium.

News media representations of homelessness : do economic news production pressures prevent journalists from adequately reporting complex social issues

This study used both content and frame analyses to test news-media representations of homelessness in The Courier-Mail newspaper for evidence of restricted journalism practice. Specifically, it sought signs of either direct manipulation of issue representation based on ideological grounds, and also evidence of news organisations prioritising low-cost news production over Public Sphere journalistic news values. The study found that news stories from the earlier parts of the longitudinal study showed stereotypical misrepresentations of homelessness for public deliberation which might be attributed to either, or both of the nominated restricting factors. However news stories from the latter part of the study saw a distinct change in the way the issue was represented, indicating a journalistic capacity to thoughtfully and sensitively represent a complex social issue to the public. Further study is recommended to ascertain how and why this change occurred, so that journalistic practice might be further improved.

Managing the production of a Cochrane systematic review

Undertaking a Cochrane systematic review can be an incredibly rewarding experience. It is however a challenging and time-consuming task. The Cochrane Handbook for Systematic Reviews of Interventions1 provides an essential resource to help reviewers navigate the often complex methodological issues of systematic review research. Additional guidelines have been developed for those undertaking reviews of public health topics,2 and Cochrane Centres throughout the world offer invaluable training opportunities. This emphasis on training and methodological rigour has helped Cochrane reviews become one of the most respected sources of synthesized research available. Even with the assistance available, however, many authors with good intentions register titles and prepare protocols but fail to publish the completed review. Data extracted from Cochrane’s Information Management System (Archie) in June 2010 showed that there were 1,301 titles registered more than two years ago that have not been published as a full review.3 Of these registered titles, 697 have had protocols published (25 are no longer active) while 604 have not even progressed to this stage (154 are no longer active). There are also 146 protocols that have been published for more than two years without being converted into completed reviews. These registered titles and protocols that have not yet progressed to a completed review represent a significant amount of time and energy invested by review authors, Cochrane editorial staff and, in some cases, external referees...

Exploring the Pro-Am interface between production and produsage

This conference celebrates the passing of 40 years since the establishment of the Internet (dating this, presumably, to the first connection between two nodes on ARPANET in October 1969). For a gathering of media scholars such as this, however, it may be just as important not only to mark the first testing of the core technologies upon which much of our present-day Net continues to build, but also to reflect on another recent milestone: the 20th anniversary of what is today arguably the chief interface through which billions around the world access and experience the Internet – the World Wide Web, launched by Tim Berners-Lee in 1989.

The potential for ethanol production from sugarcane in Australia

The Australian sugar industry processes approximately 35 million tonnes of sugarcane per year from 400 000 hectares of land. Sugar remains the principal revenue stream from sugarcane in Australia with less than 60 ML/y of fuel ethanol produced from final molasses at present. Modelling has been undertaken to estimate the potential ethanol production from the Australian sugar industry for integrated facilities producing both sugar and ethanol from the entire sugarcane resource. Although research aimed at developing commercial processes is ongoing, the use of a proportion of the bagasse and trash for ethanol production, in addition to juice and molasses fermentation, would allow significant increases in the scale of ethanol production from sugarcane in Australia, increasing total industry revenues while maintaining energy self sufficiency.

Optimising preventive maintenance strategy for production Lines

Preventive Maintenance (PM) is often applied to improve the reliability of production lines. A Split System Approach (SSA) based methodology is presented to assist in making optimal PM decisions for serial production lines. The methodology treats a production line as a complex series system with multiple (imperfect) PM actions over multiple intervals. The conditional and overall reliability of the entire production line over these multiple PM intervals are hierarchically calculated using SSA, and provide a foundation for cost analysis. Both risk-related cost and maintenance-related cost are factored into the methodology as either deterministic or random variables. This SSA based methodology enables Asset Management (AM) decisions to be optimised considering a variety of factors including failure probability, failure cost, maintenance cost, PM performance, and the type of PM strategy. The application of this new methodology and an evaluation of the effects of these factors on PM decisions are demonstrated using an example. The results of this work show that the performance of a PM strategy can be measured by its Total Expected Cost Index (TECI). The optimal PM interval is dependent on TECI, PM performance and types of PM strategies. These factors are interrelated. Generally, it was found that a trade-off between reliability and the number of PM actions needs to be made so that one can minimise Total Expected Cost (TEC) for asset maintenance.

"Who Are You" by DeepBlue Orchestra - Production of Recordings

The development and recording of 10 songs for a CD to accompany DeepBlue's new live orchestra production "Who Are You" which began touring Australia and Asia in 2012.

From ABCs to ADHD : the role of schooling the construction of behaviour disorder and the production of disorderly objects

Discussion of Attention-Deficit/Hyperactivity Disorder (ADHD) in the media, and thus much popular discourse, typically revolves around the possible causes of disruptive behaviour and the “behaviourally disordered” child. The usual suspects - too much television and video games, food additives, bad parenting, lack of discipline and single mothers – feature prominently as potential contributors to the spiralling rate of ADHD diagnosis in Western industrialised nations, especially the United States and Australia. Conspicuously absent from the field of investigation, however, is the scene of schooling and the influence that the discourses and practices of schooling might bring to bear upon the constitution of “disorderly behaviour” and subsequent recognition of particular children as a particular kind of “disorderly”. This paper reviews a sample of the literature surrounding ADHD, in order to question the function of this absence and, ultimately, make an argument for an interrogation of the school as a site for the production of disorderly objects.