The proximal first exon architecture of the murine ghrelin gene is highly similar to its human orthologue

BACKGROUND: The murine ghrelin gene (Ghrl), originally sequenced from stomach tissue, contains five exons and a single transcription start site in a short, 19 bp first exon (exon 0). We recently isolated several novel first exons of the human ghrelin gene and found evidence of a complex transcriptional repertoire. In this report, we examined the 5' exons of the murine ghrelin orthologue in a range of tissues using 5' RACE. -----FINDINGS: 5' RACE revealed two transcription start sites (TSSs) in exon 0 and four TSSs in intron 0, which correspond to 5' extensions of exon 1. Using quantitative, real-time RT-PCR (qRT-PCR), we demonstrated that extended exon 1 containing Ghrl transcripts are largely confined to the spleen, adrenal gland, stomach, and skin. -----CONCLUSION: We demonstrate that multiple transcription start sites are present in exon 0 and an extended exon 1 of the murine ghrelin gene, similar to the proximal first exon organisation of its human orthologue. The identification of several transcription start sites in intron 0 of mouse ghrelin (resulting in an extension of exon 1) raises the possibility that developmental-, cell- and tissue-specific Ghrl mRNA species are created by employing alternative promoters and further studies of the murine ghrelin gene are warranted.

The importance of gestation process dynamics

Principal Topic In this paper we seek to highlight the important intermediate role that the gestation process plays in entrepreneurship by examining its key antecedents and its consequences for new venture emergence. In doing so we take a behavioural perspective and argue that it is not only what a nascent venture is, but what it does (Katz & Gartner, 1988; Shane & Delmar, 2004; Reynolds, 2007) and when it does it during start-up (Reynolds & Miller, 1992; Lichtenstein, Carter, Dooley & Gartner, 2007) that is important. To extend an analogy from biological development, what we suggest is that the way a new venture is nurtured is just as fundamental as its nature. Much prior research has focused on the nature of new ventures and attempted to attribute variations in outcomes directly to the impact resource endowments and investments have. While there is little doubt that venture resource attributes such as human capital, and specifically prior entrepreneurial experience (Alsos & Kolvereid, 1998), access to social (Davidsson & Honig, 2003) and financial capital have an influence. Resource attributes themselves are distal from successful start-up endeavours and remain inanimate if not for the actions of the nascent venture. The key contribution we make is to shift focus from whether or not actions are taken, but when these actions happen and how that is situated in the overall gestation process. Thus, we suggest that it is gestation process dynamics, or when gestation actions occur, that is more proximal to venture outcomes and we focus on this. Recently scholars have highlighted the complexity that exists in the start-up or gestation process, be it temporal or contextual (Liao, Welsch & Tan, 2005; Lichtenstein et al. 2007). There is great variation in how long a start-up process might take (Reynolds & Miller, 1992), some processes require less action than others (Carter, Gartner & Reynolds, 1996), and the overall intensity of the start-up effort is also deemed important (Reynolds, 2007). And, despite some evidence that particular activities are more influential than others (Delmar & Shane, 2003), the order in which events may happen is, until now, largely indeterminate as regard its influence on success (Liao & Welsch, 2008). We suggest that it is this complexity of the intervening gestation process that attenuates the effect of resource endowment and has resulted in mixed findings in previous research. Thus, in order to reduce complexity we shall take a holistic view of the gestation process and argue that it is its’ dynamic properties that determine nascent venture attempt outcomes. Importantly, we acknowledge that particular gestation processes of themselves would not guarantee successful start-up, but it is more correctly the fit between the process dynamics and the ventures attributes (Davidsson, 2005) that is influential. So we aim to examine process dynamics by comparing sub-groups of venture types by resource attributes. Thus, as an initial step toward unpacking the complexity of the gestation process, this paper aims to establish the importance of its role as an intermediary between attributes of the nascent venture and the emergence of that venture. Here, we make a contribution by empirically examining gestation process dynamics and their fit with venture attributes. We do this by firstly, examining that nature of the influence that venture attributes such as human and social capital have on the dynamics of the gestation process, and secondly by investigating the effect that gestation process dynamics have on venture creation outcomes. Methodology and Propositions In order to explore the importance that gestation processes dynamics have in nascent entrepreneurship we conduct an empirical study of ventures start-ups. Data is drawn from a screened random sample of 625 Australian nascent business ventures prior to them achieving consistent outcomes in the market. This data was collected during 2007/8 and 2008/9 as part of the Comprehensive Australian Study of Entrepreneurial Emergence (CAUSEE) project (Davidsson et al., 2008). CAUSEE is a longitudinal panel study conducted over four years, sourcing information from annually administered telephone surveys. Importantly for our study, this methodology allows for the capture and tracking of active nascent venture creation as it happens, thus reducing hindsight and selection biases. In addition, improved tests of causality may be made given that outcome measures are temporally removed from preceding events. The data analysed in this paper represents the first two of these four years, and for the first time has access to follow-up outcome measures for these venture attempts: where 260 were successful, 126 were abandoned, and 191 are still in progress. With regards to venture attributes as gestation process antecedents, we examine specific human capital measured as successful prior experience in entrepreneurship, and direct social capital of the venture as ‘team start-ups’. In assessing gestation process dynamics we follow Lichtenstein et al. (2007) to suggest that the rate, concentration and timing of gestation activities may be used to summarise the complexity dynamics of that process. In addition, we extend this set of measures to include the interaction of discovery and exploitation by way of changes made to the venture idea. Those ventures with successful prior experience or those who conduct symbiotic parallel start-up attempts may be able to, or be forced to, leave their gestation action until later and still derive a successful outcome. In addition access to direct social capital may provide the support upon which the venture may draw in order to persevere in the face of adversity, turning a seemingly futile start-up attempt into a success. On the other hand prior experience may engender the foresight to terminate a venture attempt early should it be seen to be going nowhere. The temporal nature of these conjectures highlight the importance that process dynamics play and will be examined in this research Statistical models are developed to examine gestation process dynamics. We use multivariate general linear modelling to analyse how human and social capital factors influence gestation process dynamics. In turn, we use event history models and stratified Cox regression to assess the influence that gestation process dynamics have on venture outcomes. Results and Implications What entrepreneurs do is of interest to both scholars and practitioners’ alike. Thus the results of this research are important since they focus on nascent behaviour and its outcomes. While venture attributes themselves may be influential this is of little actionable assistance to practitioners. For example it is unhelpful to say to the prospective first time entrepreneur “you’ll be more successful if you have lots of prior experience in firm start-ups”. This research attempts to close this relevance gap by addressing what gestation behaviours might be appropriate, when actions best be focused, and most importantly in what circumstances. Further, we make a contribution to the entrepreneurship literature, examining the role that gestation process dynamics play in outcomes, by specifically attributing these to the nature of the venture itself. This extension is to the best of our knowledge new to the research field.

The comprehensive Australian Study of entrepreneurial emergence (CAUSEE) high potential nascent entrepreneurs : some preliminary findings

Principal Topic The Comprehensive Australian Study of Entrepreneurial Emergence (CAUSEE) represents the first Australian study to employ and extend the longitudinal and large scale systematic research developed for the Panel Study of Entrepreneurial Dynamics (PSED) in the US (Gartner, Shaver, Carter and Reynolds, 2004; Reynolds, 2007). This research approach addresses several shortcomings of other data sets including under coverage; selection bias; memory decay and hindsight bias, and lack of time separation between the assessment of causes and their assumed effects (Johnson et al 2006; Davidsson 2006). However, a remaining problem is that any a random sample of start-ups will be dominated by low potential, imitative ventures. In recognition of this issue CAUSEE supplemented PSED-type random samples with theoretically representative samples of the 'high potential' emerging ventures employing a unique methodology using novel multiple screening criteria. We define new ''high-potential'' ventures as new entrepreneurial innovative ventures with high aspirations and potential for growth. This distinguishes them from those ''lifestyle'' imitative businesses that start small and remain intentionally small (Timmons, 1986). CAUSEE is providing the opportunity to explore, for the first time, if process and outcomes of high potentials differ from those of traditional lifestyle firms. This will allows us to compare process and outcome attributes of the random sample with the high potential over sample of new firms and young firms. The attributes in which we will examine potential differences will include source of funding, and internationalisation. This is interesting both in terms of helping to explain why different outcomes occur but also in terms of assistance to future policymaking, given that high growth potential firms are increasingly becoming the focus of government intervention in economic development policies around the world. The first wave of data of a four year longitudinal study has been collected using these samples, allowing us to also provide some initial analysis on which to continue further research. The aim of this paper therefore is to present some selected preliminary results from the first wave of the data collection, with comparisons of high potential with lifestyle firms. We expect to see owing to greater resource requirements and higher risk profiles, more use of venture capital and angel investment, and more internationalisation activity to assist in recouping investment and to overcome Australia's smaller economic markets Methodology/Key Propositions In order to develop the samples of 'high potential' in the NF and YF categories a set of qualification criteria were developed. Specifically, to qualify, firms as nascent or young high potentials, we used multiple, partly compensating screening criteria related to the human capital and aspirations of the founders as well as the novelty of the venture idea, and venture high technology. A variety of techniques were also employed to develop a multi level dataset of sources to develop leads and firm details. A dataset was generated from a variety of websites including major stakeholders including the Federal and State Governments, Australian Chamber of Commerce, University Commercialisation Offices, Patent and Trademark Attorneys, Government Awards and Industry Awards in Entrepreneurship and Innovation, Industry lead associations, Venture Capital Association, Innovation directories including Australian Technology Showcase, Business and Entrepreneurs Magazines including BRW and Anthill. In total, over 480 industry, association, government and award sources were generated in this process. Of these, 74 discrete sources generated high potentials that fufilled the criteria. 1116 firms were contacted as high potential cases. 331 cases agreed to participate in the screener, with 279 firms (134 nascents, and 140 young firms) successfully passing the high potential criteria. 222 Firms (108 Nascents and 113 Young firms) completed the full interview. For the general sample CAUSEE conducts screening phone interviews with a very large number of adult members of households randomly selected through random digit dialing using screening questions which determine whether respondents qualify as 'nascent entrepreneurs'. CAUSEE additionally targets 'young firms' those that commenced trading from 2004 or later. This process yielded 977 Nascent Firms (3.4%) and 1,011 Young Firms (3.6%). These were directed to the full length interview (40-60 minutes) either directly following the screener or later by appointment. The full length interviews were completed by 594 NF and 514 YF cases. These are the cases we will use in the comparative analysis in this report. Results and Implications The results for this paper are based on Wave one of the survey which has been completed and the data obtained. It is expected that the findings will assist in beginning to develop an understanding of high potential nascent and young firms in Australia, how they differ from the larger lifestyle entrepreneur group that makes up the vast majority of the new firms created each year, and the elements that may contribute to turning high potential growth status into high growth realities. The results have implications for Government in the design of better conditions for the creation of new business, firms who assist high potentials in developing better advice programs in line with a better understanding of their needs and requirements, individuals who may be considering becoming entrepreneurs in high potential arenas and existing entrepreneurs make better decisions.

Twisted Edwards curves revisited

This paper introduces fast algorithms for performing group operations on twisted Edwards curves, pushing the recent speed limits of Elliptic Curve Cryptography (ECC) forward in a wide range of applications. Notably, the new addition algorithm uses for suitably selected curve constants. In comparison, the fastest point addition algorithms for (twisted) Edwards curves stated in the literature use . It is also shown that the new addition algorithm can be implemented with four processors dropping the effective cost to . This implies an effective speed increase by the full factor of 4 over the sequential case. Our results allow faster implementation of elliptic curve scalar multiplication. In addition, the new point addition algorithm can be used to provide a natural protection from side channel attacks based on simple power analysis (SPA).

Jacobi quartic curves revisited

This paper provides new results about efficient arithmetic on Jacobi quartic form elliptic curves, y 2 = d x 4 + 2 a x 2 + 1. With recent bandwidth-efficient proposals, the arithmetic on Jacobi quartic curves became solidly faster than that of Weierstrass curves. These proposals use up to 7 coordinates to represent a single point. However, fast scalar multiplication algorithms based on windowing techniques, precompute and store several points which require more space than what it takes with 3 coordinates. Also note that some of these proposals require d = 1 for full speed. Unfortunately, elliptic curves having 2-times-a-prime number of points, cannot be written in Jacobi quartic form if d = 1. Even worse the contemporary formulae may fail to output correct coordinates for some inputs. This paper provides improved speeds using fewer coordinates without causing the above mentioned problems. For instance, our proposed point doubling algorithm takes only 2 multiplications, 5 squarings, and no multiplication with curve constants when d is arbitrary and a = ±1/2.

Faster group operations on elliptic curves

This paper improves implementation techniques of Elliptic Curve Cryptography. We introduce new formulae and algorithms for the group law on Jacobi quartic, Jacobi intersection, Edwards, and Hessian curves. The proposed formulae and algorithms can save time in suitable point representations. To support our claims, a cost comparison is made with classic scalar multiplication algorithms using previous and current operation counts. Most notably, the best speeds are obtained from Jacobi quartic curves which provide the fastest timings for most scalar multiplication strategies benefiting from the proposed 12M + 5S + 1D point doubling and 7M + 3S + 1D point addition algorithms. Furthermore, the new addition algorithm provides an efficient way to protect against side channel attacks which are based on simple power analysis (SPA). Keywords: Efficient elliptic curve arithmetic,unified addition, side channel attack.

The genetic basis of human height : the role of estrogen

Height is a complex physical trait that displays strong heritability. Adult height is related to length of the long bones, which is determined by growth at the epiphyseal growth plate. Longitudinal bone growth occurs via the process of endochondral ossification, where bone forms over the differentiating cartilage template at the growth plate. Estrogen plays a major role in regulating longitudinal bone growth and is responsible for inducing the pubertal growth spurt and fusion of the epiphyseal growth plate. However, the mechanism by which estrogen promotes epiphyseal fusion is poorly understood. It has been hypothesised that estrogen functions to regulate growth plate fusion by stimulating chondrocyte apoptosis, angiogenesis and bone cell invasion in the growth plate. Another theory has suggested that estrogen exposure exhausts the proliferative capacity of growth plate chondrocytes, which accelerates the process of chondrocyte senescence, leading to growth plate fusion. The overall objective of this study was to gain a greater understanding of the molecular mechanisms behind estrogen-mediated growth and height attainment by examining gene regulation in chondrocytes and the role of some of these genes in normal height inheritance. With the heritability of height so well established, the initial hypothesis was that genetic variation in candidate genes associated with longitudinal bone growth would be involved in normal adult height variation. The height-related genes FGFR3, CBFA1, ER and CBFA1 were screened for novel polymorphisms using denaturing HPLC and RFLP analysis. In total, 24 polymorphisms were identified. Two SNPs in ER (rs3757323 C>T and rs1801132 G>C) were strongly associated with adult male height and displayed an 8 cm and 9 cm height difference between homozygous genotypes, respectively. The TC haplotype of these SNPs was associated with a 6 cm decrease in height and remarkably, no homozygous carriers of the TC haplotype were identified in tall subjects. No significant associations with height were found for polymorphisms in the FGFR3, CBFA1 or VDR genes. In the epiphyseal growth plate, chondrocyte proliferation, matrix synthesis and chondrocyte hypertrophy are all major contributors to long bone growth. As estrogen plays such a significant role in both growth and final height attainment, another hypothesis of this study was that estrogen exerted its effects in the growth plate by influencing chondrocyte proliferation and mediating the expression of chondrocyte marker genes. The examination of genes regulated by estrogen in chondrocyte-like cells aimed to identify potential regulators of growth plate fusion, which may further elucidate mechanisms involved in the cessation of linear growth. While estrogen did not dramatically alter the proliferation of the SW1353 cell line, gene expression experiments identified several estrogen regulated genes. Sixteen chondrocyte marker genes were examined in response to estrogen concentrations ranging from 10-12 M to 10-8 M over varying time points. Of the genes analysed, IHH, FGFR3, collagen II and collagen X were not readily detectable and PTHrP, GHR, ER, BMP6, SOX9 and TGF1 mRNAs showed no significant response to estrogen treatments. However, the expression of MMP13, CBFA1, BCL-2 and BAX genes were significantly decreased. Interestingly, the majority of estrogen regulated genes in SW1353 cells are expressed in the hypertrophic zone of the growth plate. Estrogen is also known to regulate systemic GH secretion and local GH action. At the molecular level, estrogen functions to inhibit GH action by negatively regulating GH signalling. GH treated SW1353 cells displayed increases in MMP9 mRNA expression (4.4-fold) and MMP13 mRNA expression (64-fold) in SW1353 cells. Increases were also detected in their respective proteins. Treatment with AG490, an established JAK2 inhibitor, blocked the GH mediated stimulation of both MMP9 and MMP13 mRNA expression. The application of estrogen and GH to SW1353 cells attenuated GH-stimulated MMP13 levels, but did not affect MMP9 levels. Investigation of GH signalling revealed that SW1353 cells have high levels of activated JAK2 and exposure to GH, estrogen, AG490 and other signalling inhibitors did not affect JAK2 phosphorylation. Interestingly, AG490 treatment dramatically decreased ERK2 signalling, although GH did stimulate ERK2 phosphorylation above control levels. AG490 also decreased CBFA1 expression, a transcription factor known to activate MMP9 and MMP13. Finally, GH and estrogen treatment increased expression of SOCS3 mRNA, suggesting that SOCS3 may regulate JAK/STAT signalling in SW1353 cells. The modulation of GH-mediated MMP expression by estrogen in SW1353 cells represents a potentially novel mechanism by which estrogen may regulate longitudinal bone growth. However, further investigation is required in order to elucidate the precise mechanisms behind estrogen and GH regulation of MMP13 expression in SW1353 cells. This study has provided additional evidence that estrogen and the ER gene are major factors in the regulation of growth and the determination of adult height. Newly identified polymorphisms in the ER gene not only contribute to our understanding of the genetic basis of human height, but may also be useful in association studies examining other complex traits. This study also identified several estrogen regulated genes and indicated that estrogen modifies the expression of genes which are primarily expressed in the hypertrophic region of the epiphyseal growth plate. Furthermore, synergistic studies incorporating GH and estrogen have revealed the ability of estrogen to attenuate the effects of GH on MMP13 expression, revealing potential pathways by which estrogen may modulate growth plate fusion, longitudinal bone growth and even arthritis.

Ghrelin gene-related peptides : multifunctional endocrine/autocrine modulators in health and disease

Ghrelin is a multi-functional peptide hormone which affects various processes including growth hormone and insulin release, appetite regulation, gut motility, metabolism and cancer cell proliferation. Ghrelin is produced in the stomach and in other normal and pathological cell types. It may act as an endocrine or autocrine/paracrine factor. The ghrelin gene encodes a precursor protein, preproghrelin, from which ghrelin and other potentially active peptides are derived by alternative mRNA splicing and/or proteolytic processing. The metabolic role of the peptide obestatin, derived from the preproghrelin C-terminal region, is controversial. However, it has direct effects on cancer cell proliferation. The regulation of ghrelin expression and the mechanisms through which the peptide products arise are unclear. We have recently re-examined the organisation of the ghrelin gene and identified several novel exons and transcripts. One transcript, which lacks the ghrelin-coding region of preproghrelin, contains the coding sequence of obestatin. Furthermore, we have identified an overlapping gene on the antisense strand of ghrelin, GHRLOS, which generates transcripts that may function as non-coding regulatory RNAs or code for novel, short bioactive peptides. The identification of these novel ghrelin-gene related transcripts and peptides raises critical questions regarding their physiological function and their role in obesity, diabetes and cancer.

Crossing boundaries: Implications of advances in basic sciences for the management of addiction.

This set of papers in this issue of "Addictive Behaviors" was presented at the 2004 'Addictions' conference, which, for the first time, was held in the Southern Hemisphere, on the Sunshine Coast of Queensland, Australia. The theme of the conference, Crossing Boundaries: Implications of Advances in Basic Sciences for the Management of Addiction, speaks for itself. The papers derive from a wide range of empirical paradigms and cover issues with relevance to the development of addiction, to the maintenance of problematic use, and to assessment, treatment, and relapse. Research from Europe and the United States is represented, as well as work from Australia. An international perspective is strongly emphasized from the initial paper by Obot, Poznyak, and Moneiro, (see record 2004-19599-015) which describes the WHO Report on the Neuroscience of Psychoactive Substance Use and Dependence, and summarises some of the report's implications for policy and practice. Hall, Carter, and Morley (see record 2004-19599-014) close the issue with a paper on the wide-ranging ethical implications of advances in neuroscience research, including issues arising from the identification of high risk for addiction, the potential for coercive pharmacotherapy, use of medications to enhance function, and risks to privacy.

Clinical significance of Mycobacterium asiaticum isolates in Queensland, Australia

Mycobacterium asiaticum was first reported as a cause of human disease in 1982, with only a few cases in the literature to date. This study aims to review the clinical significance of M. asiaticum isolates in Queensland, Australia. A retrospective review (1989 to 2008) of patients with M. asiaticum isolates was conducted. Data were collected through the Queensland TB Control Centre database. Disease was defined in accordance with the American Thoracic Society criteria. Twenty-four patients (13 female) had a positive culture of M. asiaticum, many residing around the Tropic of Capricorn. M. asiaticum was responsible for pulmonary disease (n = 2), childhood lymphadenitis (n = 1), olecranon bursitis (n = 1), 6 cases of possible pulmonary disease, and 2 possible wound infections. Chronic lung disease was a risk factor for pulmonary infection, and wounds/lacerations were a risk factor for extrapulmonary disease. Extrapulmonary disease responded to local measures. Pulmonary disease responded to ethambutol-isoniazid-rifampin plus pyrazinamide for the first 2 months in one patient, and amikacin-azithromycin-minocycline in another patient. While M. asiaticum is rare in Queensland, there appears to be an environmental niche. Although often a colonizer, it can be a cause of pulmonary and extrapulmonary disease. Treatment of pulmonary disease remains challenging. Extrapulmonary disease does not mandate specific nontuberculous mycobacterium (NTM) treatment.

Comparison of methods for processing drinking water samples for the isolation of Myocbacterium avium and intracellulare

Several protocols for isolation of mycobacteria from water exist, but there is no established standard method. This study compared methods of processing potable water samples for the isolation of Mycobacterium avium and Mycobacterium intracellulare using spiked sterilized water and tap water decontaminated using 0.005% cetylpyridinium chloride (CPC). Samples were concentrated by centrifugation or filtration and inoculated onto Middlebrook 7H10 and 7H11 plates and Lowenstein-Jensen slants and into mycobacterial growth indicator tubes with or without polymyxin, azlocillin, nalidixic acid, trimethoprim, and amphotericin B. The solid media were incubated at 32°C, at 35°C, and at 35°C with CO2 and read weekly. The results suggest that filtration of water for the isolation of mycobacteria is a more sensitive method for concentration than centrifugation. The addition of sodium thiosulfate may not be necessary and may reduce the yield. Middlebrook M7H10 and 7H11 were equally sensitive culture media. CPC decontamination, while effective for reducing growth of contaminants, also significantly reduces mycobacterial numbers. There was no difference at 3 weeks between the different incubation temperatures.

Two Thai teachers' perceptions of cooperative learning

Over the past decade, the need for educational reform in Thailand has become increasingly apparent. The strong tradition of teacher-directed instruction has been the major teaching and learning style in Thai education. Recent policy changes have focused on expanding education opportunities, the implementation of compulsory education and also on more student-centered pedagogies (Carter, 2006). A suggested way of bringing about the changes necessary to improve the standard of education in primary schools and retention rates is to incorporate cooperative learning into Thai primary school classrooms. The action research in this study proceeded in four stages. There were pre-implementation interviews of teachers, teacher preparation, implementation of program, and post-implementation evaluation. This study focused on the two Thai teachers' perceptions about the benefits of cooperative learning and the roles of the teachers in cooperative learning classroom. The study found that the two teachers had constructed different levels of perceptions about the benefits of cooperative learning and the roles of the teacher. These findings indicated that Thai teachers who are relatively inexperienced, lacking in confidence and with little or no experience with group work strategies such as peer tutoring need alternative, less intense professional development programs conducted over a period of a semester than the professional development program implemented in this study. In addition to being conducted over a longer period of time, the findings also indicate that alternative teacher professional programs need to provide additional workshops to improve their understanding of cooperative learning and the teachers' roles.