A fission yeast homolog of Int-6, the mammalian oncoprotein and eIF3 subunit, induces drug resistance when overexpressed

Through a screen to identify genes that induce multi-drug resistance when overexpressed, we have identified a fission yeast homolog of Int-6, a component of the human translation initiation factor eIF3. Disruption of the murine Int-6 gene by mouse mammary tumor virus (MMTV) has been implicated previously in tumorigenesis, although the underlying mechanism is not yet understood. Fission yeast Int6 was shown to interact with other presumptive components of eIF3 in vivo, and was present in size fractions consistent with its incorporation into a 43S translation preinitiation complex. Drug resistance induced by Int6 overexpression was dependent on the AP-1 transcription factor Pap1, and was associated with increased abundance of Pap1-responsive mRNAs, but not with Pap1 relocalization. Fission yeast cells lacking the int6 gene grew slowly. This growth retardation could be corrected by the expression of full length Int6 of fission yeast or human origin, or by a C-terminal fragment of the fission yeast protein that also conferred drug resistance, but not by truncated human Int-6 proteins corresponding to the predicted products of MMTV-disrupted murine alleles. Studies in fission yeast may therefore help to explain the ways in which Int-6 function can be perturbed during MMTV-induced mammary tumorigenesis.

Full-time is a given here : part-time versus full-time job quality

This study explores full-time workers' understanding of and assumptions about part-time work against six job quality components identified in recent literature. Forty interviews were conducted with employees in a public sector agency in Australia, a study context where part-time work is ostensibly 'good quality' and is typically long term, voluntary, involving secure contracts (i.e. permanent rather than casual) and having predictable hours distributed evenly over the week and year. Despite strong collective bargaining arrangements as well as substantial legal and industrial obligations, the findings revealed some serious concerns for part-time job quality. These concerns included reduced responsibilities and lesser access to high status roles and projects, a lack of access to promotion opportunities, increased work intensity and poor workplace support. The highly gendered, part-time labour market also means that it is women who disproportionately experience this disadvantage. To foster equity, greater attention needs to focus on monitoring and enhancing job quality, regardless of hours worked.

Modulation of the SHBG signalling axis

Sex hormone-binding globulin (SHBG) is a homodimeric plasma glycoprotein that is the major sex steroid carrier-protein in the bloodstream and functions also as a key regulator of steroid bioavailability within target tissues, such as the prostate. Additionally, SHBG binds to prostatic cell membranes via the putative and unidentified SHBG receptor (RSHBG), activating a signal transduction pathway implicated in stimulating both proliferation and expression of prostate specific antigen (PSA) in prostate cell lines in vitro. A yeast-two hybrid assay suggested an interaction between SHBG and kallikrein-related protease (KLK) 4, which is a serine protease implicated in the progression of prostate cancer. The potential interaction between these two proteins was investigated in this PhD thesis to determine whether SHBG is a proteolytic substrate of KLK4 and other members of the KLK family including KLK3/PSA, KLK7 and KLK14. Furthermore, the effects from SHBG proteolytic degradation on SHBG-regulated steroid bioavailability and the activation of the putative RSHBG signal transduction pathway were examined in the LNCaP prostate cancer cell line. SHBG was found to be a proteolytic substrate of the trypsin-like KLK4 and KLK14 in vitro, yielding several proteolysis fragments. Both chymotrypsin-like PSA and KLK7 displayed insignificant proteolytic activity against SHBG. The kinetic parameters of SHBG proteolysis by KLK4 and KLK14 demonstrate a strong enzyme-substrate binding capacity, possessing a Km of 1.2 ± 0.7 µM and 2.1 ± 0.6 µM respectively. The catalytic efficiencies (kcat/Km) of KLK4 and KLK14 proteolysis of SHBG were 1.6 x 104 M-1s-1 and 3.8 x 104 M-1s-1 respectively, which were comparable to parameters previously reported for peptide substrates. N-terminal sequencing of the fragments revealed cleavage near the junction of the N- and C-terminal laminin globulin-like (G-like) domains of SHBG, resulting in the division of the two globulins and ultimately the full degradation of these fragments by KLK4 and KLK14 over time. Proteolytic fragments that may retain steroid binding were rapidly degraded by both proteases, while fragments containing residues beyond the steroid binding pocket were less degraded over the same period of time. Degradation of SHBG was inhibited by the divalent metal cations calcium and zinc for KLK4, and calcium, zinc and magnesium for KLK14. The human secreted serine protease inhibitors (serpins), α1-antitrypsin and α2-antiplasmin, inhibited KLK4 and KLK14 proteolysis of SHBG; α1-antichymotrypsin inhibited KLK4 but not KLK14 activity. The inhibition by these serpins was comparable and in some cases more effective than general trypsin protease inhibitors such as aprotinin and phenylmethanesulfonyl fluoride (PMSF). The binding of 5α-dihydrotestosterone (DHT) to SHBG modulated interactions with KLK4 and KLK14. Steroid-free SHBG was more readily digested by both enzymes than DHT-bound SHBG. Moreover, a binding interaction exists between SHBG and pro-KLK4 and pro-KLK14, with DHT strengthening the binding to pro-KLK4 only. The inhibition of androgen uptake by cultured prostate cancer cells, mediated by SHBG steroid-binding, was examined to assess whether SHBG proteolysis by KLK4 and KLK14 modulated this process. Proteolytic digestion eliminated the ability of SHBG to inhibit the uptake of DHT from conditioned media into LNCaP cells. Therefore, the proteolysis of SHBG by KLK4 and KLK14 increased steroid bioavailability in vitro, leading to an increased uptake of androgens by prostate cancer cells. Interestingly, different transcriptional responses of PSA and KLK2, which are androgen-regulated genes, to DHT-bounsd SHBG treatment were observed between low and high passage number LNCaP cells (lpLNCaP and hpLNCaP respectively). HpLNCaP cells treated with DHT-bound SHBG demonstrated a significant synergistic upregulation of PSA and KLK2 above DHT or SHBG treatment alone, which is similar to previously reported downstream responses from RSHBG-mediated signaling activation. As this result was not seen in lpLNCaP cells, only hpLNCaP cells were further investigated to examine the modulation of potential RSHBG activity by KLK4 and KLK14 proteolysis of SHBG. Contrary to reported results, no increase in intracellular cAMP was observed in hpLNCaP cells when treated with SHBG in the presence and absence of either DHT or estradiol. As a result, the modulation of RSHBG-mediated signaling activation could not be determined. Finally, the identification of the RSHBG from both breast (MCF-7) and prostate cancer (LNCaP) cell lines was attempted. Fluorescently labeled peptides corresponding to the putative receptor binding domain (RBD) of SHBG were shown to be internalized by MCF-7 cells. Crosslinking of the RBD peptide to the cell surfaces of both MCF-7 and LNCaP cells, demonstrated the interaction of the peptide with several targets. These targets were then captured using RBD peptides synthesized onto a hydrophilic scaffold and analysed by mass spectrometry. The samples captured by the RBD peptide returned statistically significantly matches for cytokeratin 8, 18 and 19 as well as microtubule-actin crosslinking factor 1, which may indicate a novel interaction between SHBG and these proteins, but ultimately failed to detect a membrane receptor potentially responsible for the putative RSHBG-mediated signaling. This PhD project has reported the proteolytic processing of SHBG by two members of the kallikrein family, KLK4 and KLK14. The effect of SHBG proteolysis by KLK4 and KLK14 on RSHBG-mediated signaling activation was unable to be determined as the reported signal transduction pathway was not activated after treatment with SHBG, in combination with either DHT or estradiol. However, the digestion of SHBG by these two proteases positively regulated androgen bioavailability to prostate cancer cells in vitro. The increased uptake of androgens is deleterious in prostate cancer due to the promotion of proliferation, metastasis, invasion and the inhibition of apoptosis. The increased bioavailability of androgens, from SHBG proteolysis by KLK4 and KLK14, may therefore promote both carcinogenesis and progression of prostate cancer. Finally, this information may contribute to the development of therapeutic treatment strategies for prostate cancer by inhibiting the proteolysis of SHBG, by KLK4 and KLK14, to prevent the increased uptake of androgens by hormone-dependent cancerous tissues.

Full scale fire tests of a new light gauge steel floor-ceiling system

Cold-formed steel members can be assembled in various combinations to provide cost-efficient and safe light gauge floor systems for buildings. Such Light gauge Steel Framing (LSF) systems are widely accepted in industrial and commercial building construction. An example application is in floor-ceiling systems. Light gauge steel floor-ceiling systems must be designed to serve as fire compartment boundaries and provide adequate fire resistance. Fire-rated floor-ceiling assemblies formed with new materials and construction methodologies have been increasingly used in buildings. However, limited research has been undertaken in the past and hence a thorough understanding of their fire resistance behaviour is not available. Recently a new composite floor-ceiling system has been developed to provide higher fire rating under standard fire conditions. But its increased fire rating could not be determined using the currently available design methods. Therefore a research project was carried out to investigate its structural and fire resistance behaviour under standard fire conditions. In this research project full scale experimental tests of the new LSF floor system based on a composite ceiling unit were undertaken using a gas furnace at the Queensland University of Technology. Both the conventional and the new steel floor-ceiling systems were tested under structural and fire loads. Full scale fire tests provided a good understanding of the fire behaviour of the LSF floor-ceiling systems and confirmed the superior performance of the new composite system. This paper presents the details of this research into the structural and fire behaviour of light gauge steel floor systems protected by the new composite panel, and the results.

Placing the creative sector in innovation : the full gamut

This paper examines the place of the creative sector -- the arts, design, media and communications -- within the framework of contemporary innovation. The historical focus on science-and-technology by innovation policy makers has spurred many within the creative sector to argue how and why it also contributes to innovation. Drawing on a wide range of English-speaking research and policy documents, the full gamut of places for the creative sector in innovation is surveyed. The paper ends by scoping out the conceptual and empirical research that is required if ideas about innovation in the creative sector are to take up a mature position within innovation studies and related policy.

The mobile Internet and small business : an exploratory study of needs, uses and adoption with full-adopters of technology

The technological environment in which contemporary small and medium-sized enterprises (SMEs) operate can only be described as dynamic. The exponential rate of technological change, characterised by perceived increases in the benefits associated with various technologies, shortening product life cycles and changing standards, provides for the SME a complex and challenging operational context. The primary aim of this research was to concentrate on those SMEs that had already adopted technology in order to identify their needs for the new mobile data technologies (MDT), the mobile Internet. The research design utilised a mixed approach whereby both qualitative and quantitative data was collected to address the question. Overall, the needs of these SMEs for MDT can be conceptualised into three areas where the technology will assist business practices; communication, eCommerce and security.

Avoiding full extension field arithmetic in pairing computations

The most costly operations encountered in pairing computations are those that take place in the full extension field Fpk . At high levels of security, the complexity of operations in Fpk dominates the complexity of the operations that occur in the lower degree subfields. Consequently, full extension field operations have the greatest effect on the runtime of Miller’s algorithm. Many recent optimizations in the literature have focussed on improving the overall operation count by presenting new explicit formulas that reduce the number of subfield operations encountered throughout an iteration of Miller’s algorithm. Unfortunately, almost all of these improvements tend to suffer for larger embedding degrees where the expensive extension field operations far outweigh the operations in the smaller subfields. In this paper, we propose a new way of carrying out Miller’s algorithm that involves new explicit formulas which reduce the number of full extension field operations that occur in an iteration of the Miller loop, resulting in significant speed ups in most practical situations of between 5 and 30 percent.

Greenhouse Gas Fluxes in Tropical and Temperae Agriculture: The Need for a Full-Cost Accounting of Global Warming Potentials

Agriculture's contribution to radiative forcing is principally through its historical release of carbon in soil and vegetation to the atmosphere and through its contemporary release of nitrous oxide (N2O) and methane (CHM4). The sequestration of soil carbon in soils now depleted in soil organic matter is a well-known strategy for mitigating the buildup of CO2 in the atmosphere. Less well-recognized are other mitigation potentials. A full-cost accounting of the effects of agriculture on greenhouse gas emissions is necessary to quantify the relative importance of all mitigation options. Such an analysis shows nitrogen fertilizer, agricultural liming, fuel use, N2O emissions, and CH4 fluxes to have additional significant potential for mitigation. By evaluating all sources in terms of their global warming potential it becomes possible to directly evaluate greenhouse policy options for agriculture. A comparison of temperate and tropical systems illustrates some of these options.

Full circle : the future of sustainable fashion manufacturing in New Zealand

Many fashion businesses in New Zealand have followed a global trend towards inexpensive off shore manufacturing. The transfer of the production of garments to overseas workers has had consequences for the wellbeing of local businesses, fashion designers and garment makers. The gradual decline of fashion manufacturing also appears to have resulted in a local fashion scene where many garments look the same in style, colour, fabric, cut and fit. The excitement of the past, where the majority of fashion designers established their own individuality through the cut and shape of the garments that they produced, may have been inadvertently lost in an effort to take advantage of cost savings achieved through mass production and manufacturing methods which are now largely unavailable in New Zealand. Consequently, a sustainable local fashion and manufacturing industry, with design integrity, seems further out of reach. This paper is focussed upon the thesis that the design and manufacture of a fashion garment, bearing in mind certain economic and practical restrictions at its inception, can contribute to a more sustainable fashion manufacturing industry in New Zealand.

Full Circle : a collection of prototypes

This exhibition engages with one of the key issues facing the fashion textiles industry in terms of future sustainability: that of the well being of fashion industry workers in Australia and New Zealand (people). This collection formed the basis of my honours dissertation (completed in New Zealand in 2008) which examines the contribution that design can make to sustainable manufacturing; particularly design for local production and consumption. An important aspect this work is the discussion of source, the work suggests that the made in China syndrome (in reference to the current state of over-consumerism in Australia and New Zealand) could be bought to a close through design to minimize waste and maximize opportunity for ‘people’: in this case both garment workers and the SMEs that employ them. The garments reflect the possibilities of focusing on a local approach that could be put into practice by a framework of SMEs that already exist. In addition the design process is highly transferrable and could be put into practice almost anywhere with minimal set up costs and a design ethos that progresses at the same pace as the skills of workers. This collection is a physical and conceptual embodiment of a source local/make local/sell local approach. The collection is an example of design that demonstrates that this is not an unrealistic ideal and is in fact possible through the development of a sustainable industry, in the sense of people, profit and planet, through adoption of a design process model that stops the waste at the source, by making better use of the raw materials and labour involved in making fashion garments. Although the focus of this research appears to centre on people and profit, this kind of source local/make local/sell local approach also has great benefits in terms of environmental sustainability.