Evidence for u.v. induction of CDKN2 mutations in melanoma cell lines

The CDKN2 gene, encoding the cyclin dependent kinase inhibitor p16, is a tumour suppressor gene involved in melanoma and maps to chromosome band 9p22. Mutations or interstitial deletions of this gene have been found both in the germline of familial melanoma cases and somatically in melanoma cell lines. Previous mutation analyses of melanoma cell lines have indicated a high frequency of C:G to T:A transitions, with all of these mutations occurring at dipyrimidine sites. Including three melanoma cell lines carrying tandem CC to TT mutations, the spectrum of mutations so far reported indicates a possible role for u.v. radiation in the mutagenesis of this gene in some tumours. To further examine this hypothesis we have characterised mutations of the CDKN2 gene in 30 melanoma cell lines. Nineteen lines carried complete or partial homozygous deletions of the gene. Of the remaining cell lines, eight were shown by direct sequencing of PCR products from exon 1 and exon 2 to carry a total of nine different mutations of CDKN2. Two cell lines carried tandem CC to TT mutations and a high rate of C:G to T:A transitions was observed. This study provides further evidence for the role of u.v. light in the genesis of melanoma, with one target being the CDKN2 tumour suppressor gene.

CDKN2A/p16 is inactivated in most melanoma cell lines

The CDKN2A gene maps to chromosome 9p21-22 and is responsible for melanoma susceptibility in some families. Its product, p16, binds specifically to CDK4 and CDK6 in vitro and in vivo, inhibiting their kinase activity. CDKN2A is homozygously deleted or mutated in a large proportion of tumor cell lines and some primary tumors, including melanomas. The aim of this study was to investigate the involvement of CDKN2A and elucidate the mechanisms of p16 inactivation in a panel of 60 cell lines derived from sporadic melanomas. Twenty-six (43%) of the melanoma lines were homozygously deleted for CDKN2A, and an additional 15 (25%) lines carried missense, nonsense, or frameshift mutations. All but one of the latter group were shown by microsatellite analysis to be hemizygous for the region of 9p surrounding CDKN2A. p16 was detected by Western blotting in only five of the cell lines carrying mutations. Immunoprecipitation of p16 in these lines, followed by Western blotting to detect the coprecipitation of CDK4 and CDK6, revealed that p16 was functionally compromised in all cell lines but the one that carried a heterozygous CDKN2A mutation. In the remaining 19 lines that carried wild-type CDKN2A alleles, Western blot analysis and immunoprecipitation indicated that 11 cell lines expressed a wild-type protein. Northern blotting was performed on the remaining eight cell lines and revealed that one cell line carried an aberrantly sized RNA transcript, and two other cell lines failed to express RNA. The promoter was found to be methylated in five cell lines that expressed CDKN2A transcript but not p16. Presumably, the message seen by Northern blotting in these cell lines is the result of cross-hybridization of the total cDNA probe with the exon 1beta transcript. Microsatellite analysis revealed that the majority of these cell lines were hemi/homozygous for the region surrounding CDKN2A, indicating that the wild-type allele had been lost. In the 11 cell lines that expressed functional p16, microsatellite analysis revealed loss of heterozygosity at the markers immediately surrounding CDKN2A in five cases, and the previously characterized R24C mutation of CDK4 was identified in one of the remaining 6 lines. These data indicate that 55 of 60 (92%) melanoma cell lines demonstrated some aberration of CDKN2A or CDK4, thus suggesting that this pathway is a primary genetic target in melanoma development.

Mutations in exon 3 of the beta-catenin gene are rare in melanoma cell lines

Mutations in exon 3 of the CTNNB1 gene encoding beta-catenin have been reported in colorectal cancer cell lines and tumours. Although one study reported mutations or deletions affecting beta-catenin in 20% of melanoma cell lines, subsequent reports detected a much lower frequency of aberrations in uncultured melanomas. To determine whether this difference in mutation frequency reflected an in vitro culturing artefact, exon 3 of CTNNB1 was screened in a panel of 62 melanoma cell lines. In addition, reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect intragenic deletions affecting exon 3. One out of 62 (1.6%) cell lines was found to carry a mutation, indicating that aberration of the Wnt-1/wingless pathway through activation of beta-catenin is a rare event, even in melanoma cell lines.

PTEN inactivation is rare in melanoma tumours but occurs frequently in melanoma cell lines

Deletions detected in cytogenetic and loss of heterozygosity (LOH) studies indicate that at least one tumour suppressor gene maps to the long arm of chromosome 10. Previous deletion mapping studies have observed LOH on 10q in about 30% of melanomas analysed. The PTEN gene, mapping to chromosome band 10q23.3, encodes a protein with both lipid and protein phosphatase activity. Somatic mutations and deletions in have been detected in a variety of cell lines and tumours, including melanoma samples. We performed mutation analyses and extensive allelic loss studies to investigate the role this gene plays in melanoma pathogenesis. We found that a total of 34 out of 57 (60%) melanoma cell lines carried hemizygous deletions of chromosome 10q encompassing the PTEN locus. A further three cell lines carried smaller deletions excluding PTEN. Inactivation of both PTEN alleles by exon-specific homozygous deletion or mutation was observed in 13 out of 57 (23%) melanoma cell lines. The mutation spectrum observed does not indicate an important role for ultraviolet radiation in the genesis of these mutations, and evidence from three cell lines supports the acquisition of PTEN aberrations in culture. Ten out of 49 (20%) matched melanoma tumour/normal samples harboured hemizygous deletions of either the whole chromosome or most of the long arm. Mutations within were detected in only one of the 10 tumours demonstrating LOH at 10q23 that were analysed. These results suggest that PTEN inactivation may be important for the propagation of melanoma cells in culture, and that another chromosome 10 tumour suppressor gene may be important for melanoma pathogenesis.

Australian story : catalysts and inhibitors in the achievement of new women professors

Women are substantially under-represented in the professoriate in Australia with a ratio of one female professor to every three male professors. This gender imbalance has been an ongoing concern with various affirmative action programs implemented in universities but to limited effect. Hence, there is a need to investigate the catalysts for and inhibitors to women’s ascent to the professoriate. This investigation focussed on women appointed to the professoriate between 2005, when a research quality assessment was first proposed, and 2008. Henceforth, these women are referred to as “New Women Professors”. The catalysts and inhibitors in these women’s careers were investigated through an electronic survey and focus group interviews. The survey was administered to new women professors (n=255) and new men professors (n=240) to enable a comparison of responses. However, only women participated in focus group discussions (n=21). An analysis of the survey and interview data revealed that the most critical catalysts for women’s advancement to the professoriate were equal employment opportunities and mentoring. Equal opportunity initiatives provided women with access to traditionally male-dominated forums. Mentoring gave women an insider perspective on the complexity of academia and the politics of the academy. The key inhibitors to women’s career advancement were negative discrimination, the culture of the boys’ club, the tension between personal and professional life, and isolation. Negative discrimination and the boys’ club are problematic because they favour men and marginalise women. The tension between personal and professional life is a particular concern for women who bear children and typically assume the major role in a family for child rearing. Isolation was a concern for both women and men with isolation appearing to increase after ascent to the professoriate. Knowledge of the significant catalysts and inhibitors provides a pragmatic way to orient universities towards redressing the gender balance in the professoriate.

The impact of high stakes testing: the Australian story

High stakes testing in Australia was introduced in 2008 by way of the National Assessment Program – Literacy and Numeracy (NAPLAN). Currently, every year all students in Years 3, 5, 7 and 9 are assessed on the same days using national tests in Reading, Writing, Language Conventions (Spelling, Grammar and Punctuation) and Numeracy. In 2010 the NAPLAN results were published on the Federal Government MySchool website. The impact of these high stakes tests on jurisdictions, school principals, parents and students is considered in this article. We draw on reported observations from the Australian Primary Principals Association during 2009–10 testing periods across the country and published Australian research on the impact of high stakes literacy and numeracy testing. We also examine alternative approaches that include the use of assessment evidence for learning improvement purposes and for accountability purposes. In considering alternatives to the current large-scale testing approach we draw on key insights from research on teacher judgement, achievement standards and social moderation in the context of national curriculum and assessment reform in support of the suggested directions forward.

Creative industry clusters in Shanghai : a success story?

In 2005 the Shanghai municipal government adopted the notion of ‘creative industries’ as part of their economic development strategy. At the same time, they officially recognized a number of ‘Creative Industry Clusters’ (CIC) in the city; over the next five years these official clusters grew to over ninety in number. The active promotion of CIC by the local state can thus been seen as central to its adoption of the creative industries agenda, in turn part of its aspiration to become a modern, global metropolis. In the first part of this paper we look at the emergence of the creative industry agenda in China, making some general observations about the need to place such policy transfer in its specific context. We suggest how this agenda might be understood in the national context of China’s economic and cultural policy development. In the second we give a critical account of the development of the creative industries agenda in Shanghai and its relationship to that for CIC. We argue that this agenda had more to do with real estate development than the promotion of a ‘creative milieu’ or ‘ecosystem’, and we also give some reasons as to why this was the case. In the third section we provide some new evidence to suggest the increasing disjunction between CIC and such a wider ‘creative milieu’. In the final section we suggest some new ways in which these CIC might be approached by local government in Shanghai .

Corona ions from high-voltage power lines : nature of emission and dispersion

Positive and negative small ions, aerosol ion and number concentration and dc electric fields were monitored at an overhead high-voltage power line site. We show that the emission of corona ions was not spatially uniform along the lines and occurred from discrete components such as a particular set of spacers. Maximum ion concentrations and atmospheric dc electric fields were observed at a point 20 m downwind of the lines. It was estimated that less than 7% of the total number of aerosol particles was charged. The electrical parameters decreased steadily with further downwind distance but remained significantly higher than background.

The story of the sixties...a pile-up on the freeway of advanced art

When does 1960s art begin and end? Certainly, aside from a few affinities, the decade’s artistic output does not exactly correspond to its popular conception as the ‘Swinging Sixties’. While it was rare that psychedelic art was truly challenging, the decade saw a number of perceptions change regarding the aims, boundaries and possibilities of experiencing art. Thus, this era has come to represent a watershed or crisis in modernist art. While in the Australian context many of these nascent trends were properly realised in the 1970s – with the full force and impact of post-object art – other challenges were first articulated in the 1950s. So, like any other demarcation of a decade, its limits and boundaries are porous.

Documentary as history, memory, dream and film

A century ago, as the Western world embarked on a period of traumatic change, the visual realism of photography and documentary film brought print and radio news to life. The vision that these new mediums threw into stark relief was one of intense social and political upheaval: the birth of modernity fired and tempered in the crucible of the Great War. As millions died in this fiery chamber and the influenza pandemic that followed, lines of empires staggered to their fall, and new geo-political boundaries were scored in the raw, red flesh of Europe. The decade of 1910 to 1919 also heralded a prolific period of artistic experimentation. It marked the beginning of the social and artistic age of modernity and, with it, the nascent beginnings of a new art form: film. We still live in the shadow of this violent, traumatic and fertile age; haunted by the ghosts of Flanders and Gallipoli and its ripples of innovation and creativity. Something happened here, but to understand how and why is not easy; for the documentary images we carry with us in our collective cultural memory have become what Baudrillard refers to as simulacra. Detached from their referents, they have become referents themselves, to underscore other, grand narratives in television and Hollywood films. The personal histories of the individuals they represent so graphically–and their hope, love and loss–are folded into a national story that serves, like war memorials and national holidays, to buttress social myths and values. And, as filmic images cross-pollinate, with each iteration offering a new catharsis, events that must have been terrifying or wondrous are abstracted. In this paper we first discuss this transformation through reference to theories of documentary and memory–this will form a conceptual framework for a subsequent discussion of the short film Anmer. Produced by the first author in 2010, Anmer is a visual essay on documentary, simulacra and the symbolic narratives of history. Its form, structure and aesthetic speak of the confluence of documentary, history, memory and dream. Located in the first decade of the twentieth century, its non-linear narratives of personal tragedy and poetic dreamscapes are an evocative reminder of the distance between intimate experience, grand narratives, and the mythologies of popular films. This transformation of documentary sources not only played out in the processes of the film’s production, but also came to form its theme.