Modulation of proliferation-specific and differentiation-specific markers in human keratinocytes by SMAD7

We examined the potential role of SMAD7 in human epidermal keratinocyte differentiation. Overexpression of SMAD7 inhibited the activity of the proliferation-specific promoters for the keratin 14 and cdc2 genes and reduced the expression of the mRNA for the proliferation-specific genes cdc2 and E2F1. The ability of SMAD7 to suppress cdc2 promoter activity was lost in transformed keratinocyte cell lines and was mediated by a domain(s) located between aa 195-395 of SMAD7. This domain lies outside the domain required to inhibit TGFbeta1 signaling, suggesting that this activity is mediated by a novel functional domain(s). Examination of AP1, NFkappaB, serum response element, Gli, wnt, and E2F responsive reporters indicated that SMAD7 significantly suppressed the E2F responsive reporter and modestly increased AP1 activity in proliferating keratinocytes. These data Suggest that SMAD7 may have a role in TGFbeta-independent signaling events in proliferating/undifferentiated keratinocytes. The effects of SMAD7 in differentiated keratinocytes indicated a more traditional role for SMAD7 as an inhibitor of TGFbeta action. SMAD7 was unable to initiate the expression of differentiation markers but was able to superinduce/derepress differentiation-specific markers and genes in differentiated keratinocytes. This latter role is consistent with the ability of SMAD7 to inhibit TGFbeta-mediated suppression of keratinocyte differentiation and suggest that the opposing actions of SMAD7 and TGFbeta may serve to modulate squamous differentiation. (C) 2004 Elsevier Inc. All rights reserved.

Transient production of recombinant proteins by chinese hamster ovary cells using polyethyleneimine/DNA complexes in combination with microtubule disrupting anti-mitotic agents

We have developed a simple and robust transient expression system utilizing the 25 kDa branched cationic polymer polyethylenimine (PEI) as a vehicle to deliver plasmid DNA into suspension-adapted Chinese hamster ovary cells synchronized in G2/M phase of the cell cycle by anti-mitotic microtubule disrupting agents. The PEI-mediated transfection process was optimized with respect to PEI nitrogen to DNA phosphate molar ratio and the plasmid DNA mass to cell ratio using a reporter construct encoding firefly luciferase. Optimal production of luciferase was observed at a PEI N to DNA P ratio of 10:1 and 5 mug DNA 10(6) cells(-1). To manipulate transgene expression at mitosis, we arrested cells in G2/M phase of the cell cycle using the microtubule depolymerizing agent nocodazole. Using secreted human alkaline phosphatase (SEAP) and enhanced green fluorescent protein (eGFP) as reporters we showed that continued inclusion of nocodazole in cell culture medium significantly increased both transfection efficiency and reporter protein production. In the presence of nocodazole, greater than 90% of cells were eGFP positive 24 h post-transfection and qSEAP was increased almost fivefold, doubling total SEAP production. Under optimal conditions for PEI-mediated transfection, transient production of a recombinant chimeric IgG(4) encoded on a single vector was enhanced twofold by nocodazole, a final yield of approximately 5 mug mL(-1) achieved at an initial viable cell density of 1 x 10(6) cells mL(-1). The glycosylation of the recombinant antibody at Asn(297) was not significantly affected by nocodazole during transient production by this method. (C) 2004 Wiley Periodicals, Inc.

Learning difficulties in numeracy in Australia

In this article, we provide an understanding of the term numeracy as it is used in Australia and a description of numeracy education in this country. In particular, we discuss the role of outcomes-based curriculum frameworks and outline the dominant teaching approaches. The focus is on students with learning difficulties and how they are identified and supported in schools. We create two vignettes based on real students with difficulties in numeracy, which highlight two of the most common problems. We report on the prevalence of learning difficulties in numeracy in Australia and describe some of the initiatives related to the assessment and enhancement of learning in numeracy that are being undertaken in various states in Australia. Finally, we identify some of the future challenges facing the Australian education system in this area. These challenges relate to teacher knowledge, mandated assessment, and the role of parents. We conclude the article by calling for the evaluation of current assessment and intervention initiatives as well as the development of a national program to support the goal of numeracy for all students.

Lifetime Subsidies in Australian Private Health Insurance Markets with Community Rating

This paper considers the problem of inducing low-risk individuals of all ages to buy private health insurance in Australia. Our proposed subsidy scheme improves upon the age-based penalty scheme under the current "Australian Lifetime Cover" (LTC) scheme. We generate an alternative subsidy profile that obviates adverse selection in private health insurance markets with mandated, age-based, community rating. Our proposal is novel in that we generate subsidies that are both risk- and age-specific, based upon actual risk probabilities. The approach we take may prove useful in other jurisdictions where the extant law mandates community rating in private health insurance markets. Furthermore, our approach is useful in jurisdictions that seek to maintain private insurance to complement existing universal public systems.

Lessons in ethnonephrology

Only recently has the nephrology community moved beyond a fairly singular focus on terminal kidney failure to embrace population-based studies of earlier stages of disease, its markers and risk factors, and of interventions. Observations in developing countries, and in minority, migrant, and disadvantaged groups in westernized countries, have promoted these developments. We are only beginning to interpret renal disease in the context of public health history, social and health transitions, changing population demography, and competing mortality. Its intimate relationships to other health issues are being progressively exposed. Perspectives on the multideterminant etiology of most disease and the pedestrian nature of most risk factors are maturing. We are challenged to reconcile epidemiologic patterns with morphology in diseased renal tissue, and to consider structural markers, such as nephron number and glomerular size, as determinants of disease susceptibility. New work force models are mandated for population-based studies and intervention programs. Intervention programs need to be integrated with other chronic disease initiatives and nested in a matrix of systematic primary care, and although flexible to changing needs, must be sustained over the long term. Cross-disciplinary collaboration is essential in designing those programs, and in promoting them to health-care funders. Substantial expansion and restructuring of the discipline is needed for the nephrology community to participate effectively in those processes.

Parental attachment and conflict behavior: Implications for offspring's attachment, loneliness, and relationship satisfaction

This study assessed the implications of parental attachment security and parental conflict behavior for offspring's relational adjustment (attachment security, loneliness, and relationship satisfaction). Further, reports of parental conflict behavior were obtained from both parents and offspring, addressing questions regarding agreement between reporters and the origin and extent of discrepant perceptions. Results revealed consistent patterns of conflict behavior and moderate agreement between reporters. However, offspring reported parental conflict behavior more negatively than parents, especially when offspring or parents were anxious about relationships. Parental attachment security had direct associations with offspring's relationship anxiety, whereas associations between parental attachment and offspring's loneliness and discomfort with closeness were mediated by parental conflict behavior. Parental conflict behavior was also associated with offspring's relationship satisfaction. The results are discussed in terms of the mechanisms involved in the intergenerational transmission of relationship difficulties.

Tailoring surface properties to build colloidal diagnostic devices: Controlling interparticle associations

The ability to control the surface properties and subsequent colloidal stability of dispersed particles has widespread applicability in many fields. Sub-micrometer fluorescent silica particles (reporters) can be used to actively encode the combinatorial synthesis of peptide libraries through interparticle association. To achieve these associations, the surface chemistry of the small fluorescent silica reporters is tailored to encourage robust adhesion to large silica microparticles onto which the peptides are synthesized. The interparticle association must withstand a harsh solvent environment multiple synthetic and washing procedures, and biological screening buffers. The encoded support beads were exposed to different solvents used for peptide synthesis, and different solutions used for biological screening including phosphate buffered saline (PBS), 2-[N-morpholino]ethane sulfonic acid (VIES) and a mixture of MES and N-(3-dimethyl-aminopropyl)-N'-ethylcarbodiimide (EDC). The number of reporters remaining adhered to the support bead was quantified after each step. The nature of the associations were explored and tested to optimize the efficiency of these phenomena. Results presented illustrate the influence of the surface functionality and polyelectrolyte modification of the reporters. These parameters were investigated through zeta potential and X-ray photoelectron spectroscopy.

Balancing curriculum processes and content in a project centred curriculum - In pursuit of graduate attributes

Chemical engineering education is challenged around the world by demands and rapid changes encompassing a wide range of technical and social drivers. Graduates must be prepared for practice in increasingly diverse workplace environments in which generic or transferable attributes such as communication and teamwork together with technical excellence are mandated by prospective employers and society at large. If academe is to successfully deliver on these graduate attributes, effective curriculum design needs to include appropriate educational processes as well as course content. Conventional teacher centred approaches, stand-alone courses and retro-fitted remedial modules have not delivered the desired outcomes. Development of the broader spectrum of attributes is more likely when students are engaged with realistic and relevant experiences that demand the integration and practice of these attributes in contexts that the students find meaningful. This paper describes and evaluates The University of Queensland's Project Centred Curriculum in Chemical Engineering (PCC), a programme-wide approach to meeting these requirements. PCC strategically integrates project-based learning with more traditional instruction. Data collected shows improved levels of student attainment of generic skills with institutional and nationally benchmarked indicators showing significant increases in student perceptions of teaching quality, and overall satisfaction with the undergraduate experience. Endorsements from Australian academic, professional and industry bodies also support the approach as more effectively aligning engineering education with professional practice requirements.

Predictors of failure by medical practitioners to report suspected child abuse in Queensland, Australia

Objective: The goal of this investigation was to examine the level of notification of child abuse and neglect and the perceived deterrents to reporting by medical practitioners, who a're mandated to report their suspicions but might choose not to do so. Design: A random sample of medical practitioners was surveyed. About three hundred medical practitioners were approached through the local Division of General Practice. 91 registered medical practitioners in Queensland, Australia, took part in the study. Results: A quarter of medical practitioners admitted failing to report suspicions, though they were mostly cognisant of their responsibility to report suspected cases of abuse and neglect. Only the belief that the suspected abuse was a single incident and unlikely to happen again predicted non-reporting (X2 [1, N =89] =7.60, p

Sustainable design practitioners: Why they must be at the centre of discussions on sustainable design education

Sustainable design education is vital for engineering students. This is to allow them to meet the challenges both engineering and the wider community will face in the future. This need has not only been mandated by Engineers Australia’s graduate attributes from an Australian perspective, but more widely the issue of sustainability is one of the greatest challenges humanity has ever faced. Engineers need to be at the forefront of this challenge, because we can not only do the greatest good, but have the potential to cause the greatest harm. The biggest question with respect to the education of engineers about sustainable design is what do engineers need to know, and how best to enable this learning. This paper argues that since the entire phenomenon of sustainable design is constantly growing and changing, it is only by looking at practitioners currently trying design sustainably, and their ways of experiencing sustainable design, can we hope to articulate what it is, and therefore what and how we need to teach engineering students. It also argues that to accommodate sustainable design within engineering, we need to go further and transform the engineering profession to enable it to meet the challenges that sustainability presents. © 2005, Australasian Association for Engineering Education