Purification and characterization of heparan sulfate from human primary osteoblasts

Heparan sulfate (HS) is a linear, highly variable, highly sulfated glycosaminoglycan sugar whose biological activity largely depends on internal sulfated domains that mediate specific binding to an extensive range of proteins. In this study we employed anion exchange chromatography, molecular sieving and enzymatic cleavage on HS fractions purified from three compartments of cultured osteoblasts-soluble conditioned media, cell surface, and extracellular matrix (ECM). We demonstrate that the composition of HS chains purified from the different compartments is structurally non-identical by a number of parameters, and that these differences have significant ramifications for their ligand-binding properties. The HS chains purified of conditioned medium had twice the binding affinity for FGF2 when compared with either cell surface or ECM HS. In contrast, similar binding of BMP2 to the three types of HS was observed. These results suggest that different biological compartments of cultured cells have structurally and functionally distinct HS species that help to modulate the flow of HS-dependent factors between the ECM and the cell surface.

Culture and creative industries in Australia

It has been argued that the origins of modern creative industries policies can be found in Australia. The Creative Nation national cultural policy statement released by the Labor government headed by the Prime Minister Paul Keating in 1994 sought an original synthesis of arts and media policies that was outwardly looking, identifying the opportunities presented by what were then new digital media technologies, and clearly stated the economic opportunities presented by promotion of what were referred to at the time as the cultural industries. Several commentators have identified the influence that Creative Nation had on the Blair Labour government when it came to power in the United Kingdom in 1997. Faced with the question of how to revitalise the once-mighty industrial cities of the U.K. after the Conservative government, the Department of Culture, Media and Sport drew upon policy documents such as Australia’s Creative Nation, as well as the experience of local governments in these cities, in looking to the cultural sectors to spearhead new jobs growth, as well as re-branding the cities as cultural or creative cities in a post-industrial economic landscape. This growing alignment of culture and economics, that has been a characteristic of creative industries policies as they have developed in Australia, Britain, East Asia and Europe, marks an interesting shift in the traditional focus of arts and cultural policy as compensatory to the economic domain. The first Chair of what would become the Arts Council of Great Britain (now the Arts Council of England) was the famous economist John Maynard Keynes. In the First Annual Report of the Arts Council for 1945-1946, prepared in the latter stages of the Second World War, Keynes proposed that “the day is not far off when the economic problem will take the back seat where it belongs, and the arena of the heart and the head will be occupied or reoccupied, by our real problems — the problems of life and of human relations, of creation and behaviour and religion”. 中文摘要 1994年工黨執政時期澳洲總理基挺（Paul Keating）發表創意的國家（The Creative Nation）的文化政策聲明堪稱是澳洲現代創意產業的起源，該聲明試圖將藝術與媒體政策結合在一起，其目的在面向海外，為新數位媒體技術尋找機會。聲明中明確指出要推動文化產業為經濟帶來機會。「文化政策也是經濟政策。文化創造財富與附加價值，對創新、行銷與設計有重要貢獻，是我們工業的標誌（badge）。我們創意的層次實際上決定了我們適應新經濟imperatives的能力。文化本身就是項重要出口，是其他產品出口的主要附件（essential accompaniment）。文化吸引觀光與學生，也是我們經濟成功之關鍵。」 創意產業的策略是構建藝術、媒體與資訊電信科技的網絡以利文化產業在國家創新政策策略中擁有一席之地。此一策略最早是由1990年代末英國布萊爾（Tony Blair）的新工黨政府所採行，其後歐洲聯盟、澳洲、紐西蘭、新加坡、台灣、南韓與中國。

Cloning and characterization of a novel human gene related to vascular endothelial growth factor

This paper describes the cloning and characterization of a new member of the vascular endothelial growth factor (VEGF) gene family, which we have designated VRF for VEGF-related-factor. Sequencing of cDNAs from a human fetal brain library and RT-PCR products from normal and tumor tissue cDNA pools indicate two alternatively spliced messages with open reading frames of 621 and 564 bp, respectively. The predicted proteins differ at their carboxyl ends resulting from a shift in the open reading frame. Both isoforms show strong homology to VEGF at their amino termini, but only the shorter isoform maintains homology to VEGF at its carboxyl terminus and conserves all 16 cysteine residues of VEGF165. Similarity comparisons of this isoform revealed overall protein identity of 48% and conservative substitution of 69% with VEGF189. VRF is predicted to contain a signal peptide, suggesting that it may be a secreted factor. The VRF gene maps to the D11S750 locus at chromosome band 11q13, and the protein coding region, spanning approximately 5 kb, is comprised of 8 exons that range in size from 36 to 431 bp. Exons 6 and 7 are contiguous and the two isoforms of VRF arise through alternate splicing of exon 6. VRF appears to be ubiquitously expressed as two transcripts of 2.0 and 5.5 kb; the level of expression is similar among normal and malignant tissues.

Compilation of somatic mutations of the CDKN2 gene in human cancers : non-random distribution of base substitutions

The CDKN2 gene, encoding the cyclin-dependent kinase inhibitor p16, is a tumour suppressor gene that maps to chromosome band 9p21-p22. The most common mechanism of inactivation of this gene in human cancers is through homozygous deletion; however, in a smaller proportion of tumours and tumour cell lines intragenic mutations occur. In this study we have compiled a database of over 120 published point mutations in the CDKN2 gene from a wide variety of tumour types. A further 50 deletions, insertions, and splice mutations in CDKN2 have also been compiled. Furthermore, we have standardised the numbering of all mutations according to the full-length 156 amino acid form of p16. From this study we are able to define several hot spots, some of which occur at conserved residues within the ankyrin domains of p16. While many of the hotspots are shared by a number of cancers, the relative importance of each position varies, possibly reflecting the role of different carcinogens in the development of certain tumours. As reported previously, the mutational spectrum of CDKN2 in melanomas differs from that of internal malignancies and supports the involvement of UV in melanoma tumorigenesis. Notably, 52% of all substitutions in melanoma-derived samples occurred at just six nucleotide positions. Nonsense mutations comprise a comparatively high proportion of mutations present in the CDKN2 gene, and possible explanations for this are discussed.

The CDKN2A (p16) gene and human cancer

CDKN2A, the gene encoding the cell-cycle inhibitor p16CDKN2A, was first identified in 1994. Since then, somatic mutations have been observed in many cancers and germline alterations have been found in kindreds with familial atypical multiple mole/melanoma (FAMMM), also known as atypical mole syndrome. In this review we tabulate the known mutations in this gene and discuss specific aspects, particularly with respect to germline mutations and cancer predisposition.

How can a mentor’s personal attributes and pedagogical knowledge develop a preservice teacher’s behaviour management?

This study presents the importance of a mentor’s (experienced teacher’s) personal attributes and pedagogical knowledge for developing a mentee’s (preservice teacher’s) teaching practices. Specifically, preservice teachers can have difficulties with behaviour management and must learn management strategies that help them to teach more effectively. This paper investigates how mentoring may facilitate the development of a mentee’s behaviour management strategies, in particular what personal attributes and pedagogical knowledge are used in this process.