Expression of PTRF in PC3 cells modulates cholesterol dynamics and the actin cytoskeleton impacting secretion pathways

Expression of caveolin-1 is up-regulated in prostate cancer metastasis and is associated with aggressive recurrence of the disease. Intriguingly, caveolin-1 is also secreted from prostate cancer cell lines and has been identified in secreted prostasomes. Caveolin-1 is the major structural component of the plasma membrane invaginations called caveolae. Co-expression of the coat protein Polymerase I and transcript release factor (PTRF) is required for caveolae formation. We recently found that expression of caveolin-1 in the aggressive prostate cancer cell line PC-3 is not accompanied by PTRF, leading to noncaveolar caveolin-1 lipid rafts. Moreover, ectopic expression of PTRF in PC-3 cells sequesters caveolin-1 into caveolae. Here we quantitatively analyzed the effect of PTRF expression on the PC-3 proteome using stable isotope labeling by amino acids in culture and subcellular proteomics. We show that PTRF reduced the secretion of a subset of proteins including secreted proteases, cytokines, and growth regulatory proteins, partly via a reduction in prostasome secretion. To determine the cellular mechanism accounting for the observed reduction in secreted proteins we analyzed total membrane and the detergent-resistant membrane fractions. Our data show that PTRF expression selectively impaired the recruitment of actin cytoskeletal proteins to the detergent-resistant membrane, which correlated with altered cholesterol distribution in PC-3 cells expressing PTRF. Consistent with this, modulating cellular cholesterol altered the actin cytoskeleton and protein secretion in PC-3 cells. Intriguingly, several proteins that function in ER to Golgi trafficking were reduced by PTRF expression. Taken together, these results suggest that the noncaveolar caveolin-1 found in prostate cancer cells generates a lipid raft microenvironment that accentuates secretion pathways, possibly at the step of ER sorting/exit. Importantly, these effects could be modulated by PTRF expression.

The Australian Consumer Law

Reform of Australia’s inconsistent Commonwealth, State and Territory consumer laws is now a reality. The 1 January 2011 commencement of the Australian Consumer Law (ACL), within the Competition and Consumer Act 2010, is the culmination of a long process of consultation. Unifying and rationalising the plethora of laws, this new Act sees the disappearance of the “Trade Practices Act” and the amendment of a raft of State and Territory legislation; the new national regime informed by them operates in their stead. This is indisputably the most comprehensive change in the history of the Trade Practices Act 1974. This book aims to assist practitioners, academics and students understand the Australian Consumer Law regime and its impact. It summarises the history and constitutional basis of the ACL, explaining how the ACL will be implemented, amended and enforced. In addition it explores how the various general and specific protections interrelate, and the scope of their overlap, and considers the content of the ACL, and the principal changes from the provisions of the Trade Practices Act.

Production of metal oxide nanoparticles

Particles having at least regions of at least one metal oxide having nano-sized grains are produced by providing particles of a material having an initial, nonequiaxed particle shape, prepg. a mixt. of these particles and at last one metal oxide precursor, and treating the mixt. such that the precursor reacts with the particles. The process can be a co-pptn. process, sol-gel synthesis, micro-emulsion method, surfactant-based process, or a process that uses polymers. Complex metal oxide nanoparticles are produced by (a) prepg. a soln. contg. metal cations, (b) mixing the soln. with a surfactant to form micelles within the soln., and (c) heating the micellar liq. to form metal oxide and to remove the surfactant. The formed metal oxide particles have essentially the same morphol. (particle size and shape) as the initial morphol. of the material particles provided. [on SciFinder(R)]

Method of making metal oxides

A method for producing particles having at least regions of at least one metal oxide having nano-sized grains comprises providing particles of material having an initial, non-equiaxed particle shape, making a mixture of the particles of material and one or more precursors of the metal oxide, and treating the mixture such that the one or more precursors of the metal oxide react with the particles of material to thereby form at least regions of metal oxide on or within the particles, wherein atoms from the particles of material form part of a matrix of the at least one metal oxide and the at least one metal oxide has nano-sized grains and wherein at least some of the regions of metal oxide on or within the particles have a non-equiaxed grain shape.

Effect of dangling chains on the structure and physical properties of a tightly crosslinked poly(ethylene glycol) network

Major imperfections in crosslinked polymers include loose or dangling chain ends that lower the crosslink d., thereby reducing elastic recovery and increasing the solvent swelling. These imperfections are hard to detect, quantify and control when the network is initiated by free radical reactions. As an alternative approach, the sol-​gel synthesis of a model poly(ethylene glycol) (PEG-​2000) network is described using controlled amts. of bis- and mono-​triethoxy silyl Pr urethane PEG precursors to give silsesquioxane (SSQ, R-​SiO1.5) structures as crosslink junctions with a controlled no. of dangling chains. The effect of the no. of dangling chains on the structure and connectivity of the dried SSQ networks has been detd. by step-​crystn. differential scanning calorimetry. The role that micelle formation plays in controlling the sol-​gel PEG network connectivity has been studied by dynamic light scattering of the bis- and mono-​triethoxy silyl precursors and the networks have been characterized by 29Si solid state NMR, sol fraction and swelling measurements. These show that the dangling chains will increase the mesh size and water uptake. Compared to other end-​linked PEG hydrogels, the SSQ-​crosslinked networks show a low sol fraction and high connectivity, which reduces solvent swelling, degree of crystallinity and the crystal transition temp. The increased degree of freedom in segment movement on the addn. of dangling chains in the SSQ-​crosslinked network facilitates the packing process in crystn. of the dry network and, in the hydrogel, helps to accommodate more water mols. before reaching equil.

Effect of chemical conjugation of L-leucine to chitosan on the dispersibility and drug release of a dry powder inhaler nanoparticle formulation

Purpose: This study investigated the effect of chemical conjugation of the amino acid L-leucine to the polysaccharide chitosan on the dispersibility and drug release pattern of a polymeric nanoparticle (NP)-based controlled release dry powder inhaler (DPI) formulation. Methods: A chemical conjugate of L-leucine with chitosan was synthesized and characterized by Infrared (IR) Spectroscopy, Nuclear Magnetic Resonance (NMR) Spectroscopy, Elemental Analysis and X-ray Photoelectron Spectroscopy (XPS). Nanoparticles of both chitosan and its conjugate were prepared by a water-in-oil emulsification – glutaraldehyde cross-linking method using the antihypertensive agent, diltiazem (Dz) hydrochloride as the model drug. The surface morphology and particle size distribution of the nanoparticles were determined by Scanning Electron Microscopy (SEM) and Dynamic Light Scattering (DLS). The dispersibility of the nanoparticle formulation was analysed by a Twin Stage Impinger (TSI) with a Rotahaler as the DPI device. Deposition of the particles in the different stages was determined by gravimetry and the amount of drug released was analysed by UV spectrophotometry. The release profile of the drug was studied in phosphate buffered saline at 37 ⁰C and analyzed by UV spectrophotometry. Results: The TSI study revealed that the fine particle fractions (FPF), as determined gravimetrically, for empty and drug-loaded conjugate nanoparticles were significantly higher than for the corresponding chitosan nanoparticles (24±1.2% and 21±0.7% vs 19±1.2% and 15±1.5% respectively; n=3, p<0.05). The FPF of drug-loaded chitosan and conjugate nanoparticles, in terms of the amount of drug determined spectrophotometrically, had similar values (21±0.7% vs 16±1.6%). After an initial burst, both chitosan and conjugate nanoparticles showed controlled release that lasted about 8 to 10 days, but conjugate nanoparticles showed twice as much total drug release compared to chitosan nanoparticles (~50% vs ~25%). Conjugate nanoparticles also showed significantly higher dug loading and entrapment efficiency than chitosan nanoparticles (conjugate: 20±1% & 46±1%, chitosan: 16±1% & 38±1%, n=3, p<0.05). Conclusion: Although L-leucine conjugation to chitosan increased dispersibility of formulated nanoparticles, the FPF values are still far from optimum. The particles showed a high level of initial burst release (chitosan, 16% and conjugate, 31%) that also will need further optimization.

Accurate measurement of the molecular thickness of thin organic shells on small inorganic cores using dynamic light scattering

Dynamic light scattering (DLS) has become a primary nanoparticle characterization technique with applications from materials characterization to biological and environmental detection. With the expansion in DLS use from homogeneous spheres to more complicated nanostructures, comes a decrease in accuracy. Much research has been performed to develop different diffusion models that account for the vastly different structures but little attention has been given to the effect on the light scattering properties in relation to DLS. In this work, small (core size < 5 nm) core-shell nanoparticles were used as a case study to measure the capping thickness of a layer of dodecanethiol (DDT) on Au and ZnO nanoparticles by DLS. We find that the DDT shell has very little effect on the scattering properties of the inorganic core and hence can be ignored to a first approximation. However, this results in conventional DLS analysis overestimating the hydrodynamic size in the volume and number weighted distributions. By introducing a simple correction formula that more accurately yields hydrodynamic size distributions a more precise determination of the molecular shell thickness is obtained. With this correction, the measured thickness of the DDT shell was found to be 7.3 ± 0.3 Å, much less than the extended chain length of 16 Å. This organic layer thickness suggests that on small nanoparticles, the DDT monolayer adopts a compact disordered structure rather than an open ordered structure on both ZnO and Au nanoparticle surfaces. These observations are in agreement with published molecular dynamics results.

Getting the right words right

Oscar Wilde once complained that having spent an entire morning putting a comma into one of his poems, he spent the afternoon removing it. Peter Selgin calls this revisionitis: the inability of a writer to part with their work, the compulsion to go on fiddling and picking until there is nothing left to fret about. There can be real danger, Selgin warns, in fussing with things. The question then of how much to revise or how little to rewrite is one without certain answer and writers themselves hold differing views...

PTRF/cavin-1 neutralizes non-caveolar caveolin-1 microdomains in prostate cancer

Caveolin-1 has a complex role in prostate cancer and has been suggested to be a potential biomarker and therapeutic target. As mature caveolin-1 resides in caveolae, invaginated lipid raft domains at the plasma membrane, caveolae have been suggested as a tumor-promoting signaling platform in prostate cancer. However, caveola formation requires both caveolin-1 and cavin-1 (also known as PTRF; polymerase I and transcript release factor). Here, we examined the expression of cavin-1 in prostate epithelia and stroma using tissue microarray including normal, non-malignant and malignant prostate tissues. We found that caveolin-1 was induced without the presence of cavin-1 in advanced prostate carcinoma, an expression pattern mirrored in the PC-3 cell line. In contrast, normal prostate epithelia expressed neither caveolin-1 nor cavin-1, while prostate stroma highly expressed both caveolin-1 and cavin-1. Utilizing PC-3 cells as a suitable model for caveolin-1-positive advanced prostate cancer, we found that cavin-1 expression in PC-3 cells inhibits anchorage-independent growth, and reduces in vivo tumor growth and metastasis in an orthotopic prostate cancer xenograft mouse model. The expression of α-smooth muscle actin in stroma along with interleukin-6 (IL-6) in cancer cells was also decreased in tumors of mice bearing PC-3-cavin-1 tumor cells. To determine whether cavin-1 acts by neutralizing caveolin-1, we expressed cavin-1 in caveolin-1-negative prostate cancer LNCaP and 22Rv1 cells. Caveolin-1 but not cavin-1 expression increased anchorage-independent growth in LNCaP and 22Rv1 cells. Cavin-1 co-expression reversed caveolin-1 effects in caveolin-1-positive LNCaP cells. Taken together, these results suggest that caveolin-1 in advanced prostate cancer is present outside of caveolae, because of the lack of cavin-1 expression. Cavin-1 expression attenuates the effects of non-caveolar caveolin-1 microdomains partly via reduced IL-6 microenvironmental function. With circulating caveolin-1 as a potential biomarker for advanced prostate cancer, identification of the molecular pathways affected by cavin-1 could provide novel therapeutic targets.

Graphene-type sheets of Nb1-xWxS2 : synthesis and in situ functionalization

Enlightened by the discovery of graphenes, a variety of inorganic analogues have been synthesized and characterized in recent years. Solvated Nb1-xWxS2 analogues of graphene-type sheets were prepared by lithiation and exfoliation of multistacked Nb1-xWxS2 coin roll nanowires (CRNWs), followed by in situ functionalization with gold nanoparticles to synthesize gold-loaded Nb1-xWxS2/Au nanocomposites. The Nb1-xWxS2 nanosheets and the corresponding Nb1-xWxS2/Au nanocomposites were characterized by high resolution electron microscopy (HRTEM), energy-dispersive X-ray spectroscopy (EDX), scanning transmission electron microscopy (STEM), dynamic light scattering (DLS) and scanning force microscopy (AFM). The graphene-type sheets are stable in water and other solvents and can be functionalized similarly as chalcogen-terminated surfaces (e.g. with Au nanoparticles).

Producing the NAPLAN Machine: A Schizoanalytic Cartography

Everything revolves around desiring-machines and the production of desire… Schizoanalysis merely asks what are the machinic, social and technical indices on a socius that open to desiring-machines (Deleuze & Guattari, 1983, pp. 380-381). Achievement tests like NAPLAN are fairly recent, yet common, education policy initiatives in much of the Western world. They intersect with, use and change pre-existing logics of education, teaching and learning. There has been much written about the form and function of these tests, the ‘stakes’ involved and the effects of their practice. This paper adopts a different “angle of vision” to ask what ‘opens’ education to these regimes of testing(Roy, 2008)? This paper builds on previous analyses of NAPLAN as a modulating machine, or a machine characterised by the increased intensity of connections and couplings. One affect can be “an existential disquiet” as “disciplinary subjects attempt to force coherence onto a disintegrating narrative of self”(Thompson & Cook, 2012, p. 576). Desire operates at all levels of the education assemblage, however our argument is that achievement testing manifests desire as ‘lack’; seen in the desire for improved results, the desire for increased control, the desire for freedom, the desire for acceptance to name a few. For Deleuze and Guattari desire is irreducible to lack, instead desire is productive. As a productive assemblage, education machines operationalise and produce through desire; “Desire is a machine, and the object of the desire is another machine connected to it”(Deleuze & Guattari, 1983, p. 26). This intersection is complexified by the strata at which they occur, the molar and molecular connections and flows they make possible. Our argument is that when attention is paid to the macro and micro connections, the machines built and disassembled as a result of high-stakes testing, a map is constructed that outlines possibilities, desires and blockages within the education assemblage. This schizoanalytic cartography suggests a new analysis of these ‘axioms’ of testing and accountability. It follows the flows and disruptions made possible as different or altered connections are made and as new machines are brought online. Thinking of education machinically requires recognising that “every machine functions as a break in the flow in relation to the machine to which it is connected, but at the same time is also a flow itself, or the production of flow, in relation to the machine connected to it”(Deleuze & Guattari, 1983, p. 37). Through its potential to map desire, desire-production and the production of desire within those assemblages that have come to dominate our understanding of what is possible, Deleuze and Guattari’s method of schizoanalysis provides a provocative lens for grappling with the question of what one can do, and what lines of flight are possible.

Education and the relation to the outside: A little real reality

One of the aims of Deleuze. Guattari. Schizoanalysis. Education. is to focus on the radical reconfiguration that education is undergoing, impacting educator, administrator, institution and ‘sector’ alike. More to the point, it is the responses to that process of reconfiguration - this newly emerging assemblage - that are a key focal point in this issue. Essential to these responses, we propose, is Deleuze and Guattari’s method of schizonalysis, which offers a way to not only understand the rules of this new game, but also, hopefully, some escape from the promise of a brave new world of continuous education and motivation. A brave new world of digitised courses, impersonal and corporate expertise, updatable performance metrics, Massive Open Online Courses (MOOCs), learning analytics, transformative teaching and learning, online high-stakes testing in the name of transforming and augmenting human capital overlays the corporeal practices of institutional surveillance, examination and categorical sorting. A brave new world, importantly, where people’s continuous education is instituted less, or not simply, through disciplinary practices, and increasingly through a constant and continuous sampling and profiling of not simply performance but their activity, measured against the profiled activity of a ‘like’ age group, person, or an institution. This continuous education, including the sampling that accompanies it, we are all informed through various information and marketing campaigns, is in our best interest. An interest that is driven and governed by an ever-increasing corporatisation and monetisation of ‘the knowledge sector’, as well as an interest that is sustained through an ever-increasing, as well as continuous, debt.

The eternal return of teaching in the time of the corporation

This article addresses the new conditions under which teachers are making the choice to teach. Our core contention is that the reorganisation of schools according to the logic of the corporation, as described in Deleuze's ‘Postscript’, is changing the flows and forces on the primary surface of ‘the classroom’. These changes block the usual movements of teaching to discipline, normalise and individualise, which was the role of the school as precursor to the factory. Blocked from repeating, or returning, teaching as it has always been done, teachers must actively re-will to teach; teachers cannot use order words to name themselves and direct flows and forces as they have usually been done. While many choices to teach will be undertaken, the most popular being that of choosing to teach toward the corporation, the repetition of teaching toward enclosed spaces becomes less compelling. Like Nietzsche's Zarathustra, teachers, who have the courage to actively choose, face a new dawn in which teaching cannot be what it once was. In that moment they must choose to repeat that choice an infinite number of times, the choice of eternal return, and it is from here that new times might begin.

Spinning in the NAPLAN ether: 'Postscript on the Control Societies' and the seduction of education in Australia

This paper applies concepts Deleuze developed in his ‘Postscript on the Societies of Control’, especially those relating to modulatory power, dividuation and control, to aspects of Australian schooling to explore how this transition is manifesting itself. Two modulatory machines of assessment, NAPLAN and My Schools, are examined as a means to better understand how the disciplinary institution is changing as a result of modulation. This transition from discipline to modulation is visible in the declining importance of the disciplinary teacher–student relationship as a measure of the success of the educative process. The transition occurs through seduction because that which purports to measure classroom quality is in fact a serpent of modulation that produces simulacra of the disciplinary classroom. The effect is to sever what happens in the disciplinary space from its representations in a luminiferous ether that overlays the classroom.