Mesenchymal stem cells

Cell based therapies as they apply to tissue engineering and regenerative medicine, require cells capable of self renewal and differentiation, and a prerequisite is to be able to prepare an effective dose of ex vivo expanded cells for autologous transplants. The in vivo identification of a source of physiologically relevant cell types suitable for cell therapies therefore figures as an integral part of tissue engineering. Stem cells serve as a reserve for biological repair, having the potential to differentiate into a number of specialised cell types within the body; they therefore represent the most useful candidates for cell based therapies. The primary goal of stem cell research is to produce cells that are both patient specific, as well as having properties suitable for the specific conditions for which they are intended to remedy. From a purely scientific perspective, stem cells allow scientists to gain a deeper understanding of developmental biology and regenerative therapies. Stem cells have acquired a number of uses for applications in regenerative medicine, immunotherapy, gene therapy, but it is in the area of tissue engineering that they generate most excitement, primarily as a result of their capacity for self-renewal and pluripotency. A unique feature of stem cells is their ability to maintain an uncommitted quiescent state in vivo and then, once triggered by conditions such as disease, injury or natural wear or tear, serve as a reservoir and natural support system to replenish lost cells. Although these cells retain the plasticity to differentiate into various tissues, being able to control this differentiation process is still one of the biggest challenges facing stem cell research. In an effort to harness the potential of these cells a number of studies have been conducted using both embryonic/foetal and adult stem cells. The use of embryonic stem cells (ESC) have been hampered by strong ethical and political concerns, this despite their perceived versatility due to their pluripotency. Ethical issues aside, other concerns raised with ESCs relates to the possibility of tumorigenesis, immune rejection and complications with immunosuppressive therapies, all of which adds layers of complications to the application ESC in research and which has led to the search for alternative sources for stem cells. The adult tissues in higher organisms harbours cells, termed adult stem cells, and these cells are reminiscent of unprogrammed stem cells. A number of sources of adult stem cells have been described. Bone marrow is by far the most accessible source of two potent populations of adult stem cells, namely haematopoietic stem cells (HSCs) and bone marrow mesenchymal stem cells (BMSCs). Autologously harvested adult stem cells can, in contrast to embryonic stem cells, readily be used in autografts, since immune rejection is not an issue; and their use in scientific research has not attracted the ethical concerns which have been the case with embryonic stem cells. The major limitation to their use, however, is the fact that adult stem cells are exceedingly rare in most tissues. This fact makes identifying and isolating these cells problematic; bone marrow being perhaps the only notable exception. Unlike the case of HSCs, there are as yet no rigorous criteria for characterizing MSCs. Changing acuity about the pluripotency of MSCs in recent studies has expanded their potential application; however, the underlying molecular pathways which impart the features distinctive to MSCs remain elusive. Furthermore, the sparse in vivo distribution of these cells imposes a clear limitation to their study in vitro. Also, when MSCs are cultured in vitro, there is a loss of the in vivo microenvironment, resulting in a progressive decline in proliferation potential and multipotentiality. This is further exacerbated with increased passage numbers in culture, characterized by the onset of senescence related changes. As a consequence, it is necessary to establish protocols for generating large numbers of MSCs but without affecting their differentiation potential. MSCs are capable of differentiating into mesenchymal tissue lineages, including bone, cartilage, fat, tendon, muscle, and marrow stroma. Recent findings indicate that adult bone marrow may also contain cells that can differentiate into the mature, nonhematopoietic cells of a number of tissues, including cells of the liver, kidney, lung, skin, gastrointestinal tract, and myocytes of heart and skeletal muscle. MSCs can readily be expanded in vitro and can be genetically modified by viral vectors and be induced to differentiate into specific cell lineages by changing the microenvironment–properties which makes these cells ideal vehicles for cellular gene therapy. MSCs can also exert profound immunosuppressive effects via modulation of both cellular and innate immune pathways, and this property allows them to overcome the issue of immune rejection. Despite the many attractive features associated with MSCs, there are still many hurdles to overcome before these cells are readily available for use in clinical applications. The main concern relates to in vivo characterization and identification of MSCs. The lack of a universal biomarker, sparse in vivo distribution, and a steady age related decline in their numbers, makes it an obvious need to decipher the reprogramming pathways and critical molecular players which govern the characteristics unique to MSCs. This book presents a comprehensive insight into the biology of adult stem cells and their utility in current regeneration therapies. The adult stem cell populations reviewed in this book include bone marrow derived MSCs, adipose derived stem cells (ASCs), umbilical cord blood stem cells, and placental stem cells. The features such as MSC circulation and trafficking, neuroprotective properties, and the nurturing roles and differentiation potential of multiple lineages have been discussed in details. In terms of therapeutic applications, the strengths of MSCs have been presented and their roles in disease treatments such as osteoarthritis, Huntington’s disease, periodontal regeneration, and pancreatic islet transplantation have been discussed. An analysis comparing osteoblast differentiation of umbilical cord blood stem cells and MSCs has been reviewed, as has a comparison of human placental stem cells and ASCs, in terms of isolation, identification and therapeutic applications of ASC in bone, cartilage regeneration, as well as myocardial regeneration. It is my sincere hope that this book will update the reader as to the research progress of MSC biology and potential use of these cells in clinical applications. It will be the best reward to all contributors of this book, if their efforts herein may in some way help the readers in any part of their study, research, and career development.

Response to "a step in the right direction : commentary on expected values for pedometer-determined physical activity in youth"

The figure Beets took exception to displays sex‐ and age‐specific median values of aggregated published expected values for pedometer determined physical activity.

Online environmental information systems

In today’s information society, electronic tools, such as computer networks for the rapid transfer of data and composite databases for information storage and management, are critical in ensuring effective environmental management. In particular environmental policies and programs for federal, state, and local governments need a large volume of up-to-date information on the quality of water, air, and soil in order to conserve and protect natural resources and to carry out meteorology. In line with this, the utilization of information and communication technologies (ICTs) is crucial to preserve and improve the quality of life. In handling tasks in the field of environmental protection a range of environmental and technical information is often required for a complex and mutual decision making in a multidisciplinary team environment. In this regard e-government provides a foundation of the transformative ICT initiative which can lead to better environmental governance, better services, and increased public participation in environmental decision- making process.

Benchmarking local E-Government

Increasingly, almost everything we do in our daily lives is being influenced by information and communications technologies (ICTs) including the Internet. The task of governance is no exception with an increasing number of national, state, and local governments utilizing ICTs to support government operations, engage citizens, and provide government services. As with other things, the process of governance is now being prefixed with an “e”. E-governance can range from simple Web sites that convey basic information to complex sites that transform the customary ways of delivering all sorts of government services. In this respect local e-government is the form of e-governance that specifically focuses on the online delivery of suitable local services by local authorities. In practice local e-government reflects four dimensions, each one dealing with the functions of government itself. The four are: (a) e-services, the electronic delivery of government information, programs, and services often over the Internet; (b) e-management, the use of information technology to improve the management of government. This might range from streamlining business processes to improving the flow of information within government departments; (c) e-democracy the use of electronic communication vehicles, such as e-mail and the Internet, to increase citizen participation in the public decision-making process; (d) e-commerce, the exchange of money for goods and services over the Internet which might include citizens paying taxes and utility bills, renewing vehicle registrations, and paying for recreation programs, or government buying office supplies and auctioning surplus equipment (Cook, LaVigne, Pagano, Dawes, & Pardo, 2002). Commensurate with the rapid increase in the process of developing e-governance tools, there has been an increased interest in benchmarking the process of local e-governance. This benchmarking, which includes the processes involved in e-governance as well as the extent of e-governance adoption or take-up is important as it allows for improved processes and enables government agencies to move towards world best practice. It is within this context that this article discusses benchmarking local e-government. It brings together a number of discussions regarding the significance of benchmarking, best practices and actions for local e-government, and key elements of a successful local e-government project.

An examination of the relationship between emotional intelligence, leadership style and perceived leadership outcomes in Australian educational institutions

In the field of leadership studies transformational leadership theory (e.g., Bass, 1985; Avolio, Bass, & Jung, 1995) has received much attention from researchers in recent years (Hughes, Ginnet, & Curphy, 2009; Hunt, 1999). Many previous studies have found that transformational leadership is related to positive outcomes such as the satisfaction, motivation and performance of followers in organisations (Judge & Piccolo, 2004; Lowe, Kroeck, & Sivasubramaniam, 1996), including in educational institutions (Chin, 2007; Leithwoood & Jantzi, 2005). Hence, it is important to explore constructs that may predict leadership style in order to identify potential transformational leaders in leadership assessment and selection procedures. Several researchers have proposed that emotional intelligence (EI) is one construct that may account for hitherto unexplained variance in transformational leadership (Mayer, 2001; Watkin, 2000). Different models of EI exist (e.g., Goleman, 1995, 2001; Bar-On, 1997; Mayer & Salovey, 1997) but momentum is growing for the Mayer and Salovey (1997) model to be considered the most useful (Ashkanasy & Daus, 2005; Daus & Ashkanasy, 2005). Studies in non-educational settings claim to have found that EI is a useful predictor of leadership style and leader effectiveness (Harms & Crede, 2010; Mills, 2009) but there is a paucity of studies which have examined the Mayer and Salovey (1997) model of EI in educational settings. Furthermore, other predictor variables have rarely been controlled in previous studies and only self-ratings of leadership behaviours, rather than multiple ratings, have usually been obtained. Therefore, more research is required in educational settings to answer the question: to what extent is the Mayer and Salovey (1997) model of EI a useful predictor of leadership style and leadership outcomes? This project, set in Australian educational institutions, was designed to move research in the field forward by: using valid and reliable instruments, controlling for other predictors, obtaining an adequately sized sample of real leaders as participants and obtaining multiple ratings of leadership behaviours. Other variables commonly used to predict leadership behaviours (personality factors and general mental ability) were assessed and controlled in the project. Additionally, integrity was included as another potential predictor of leadership behaviours as it has previously been found to be related to transformational leadership (Parry & Proctor-Thomson, 2002). Multiple ratings of leadership behaviours were obtained from each leader and their supervisors, peers and followers. The following valid and reliable psychological tests were used to operationalise the variables of interest: leadership styles and perceived leadership outcomes (Multifactor Leadership Questionnaire, Avolio et al., 1995), EI (Mayer–Salovey–Caruso Emotional Intelligence Test, Mayer, Salovey, & Caruso, 2002), personality factors (The Big Five Inventory, John, Donahue, & Kentle, 1991), general mental ability (Wonderlic Personnel Test-Quicktest, Wonderlic, 2003) and integrity (Integrity Express, Vangent, 2002). A Pilot Study (N = 25 leaders and 75 raters) made a preliminary examination of the relationship between the variables included in the project. Total EI, the experiential area, and the managing emotions and perceiving emotions branches of EI, were found to be related to transformational leadership which indicated that further research was warranted. In the Main Study, 144 leaders and 432 raters were recruited as participants to assess the discriminant validity of the instruments and examine the usefulness of EI as a predictor of leadership style and perceived leadership outcomes. Scores for each leadership scale across the four rating levels (leaders, supervisors, peers and followers) were aggregated with the exception of the management-by-exception active scale of transactional leadership which had an inadequate level of interrater agreement. In the descriptive and measurement component of the Main Study, the instruments were found to demonstrate adequate discriminant validity. The impact of role and gender on leadership style and EI were also examined, and females were found to be more transformational as leaders than males. Females also engaged in more contingent reward (transactional leadership) behaviours than males, whilst males engaged in more passive/avoidant leadership behaviours than females. In the inferential component of the Main Study, multiple regression procedures were used to examine the usefulness of EI as a predictor of leadership style and perceived leadership outcomes. None of the EI branches were found to be related to transformational leadership or the perceived leadership outcomes variables included in the study. Openness, emotional stability (the inverse of neuroticism) and general mental ability (inversely) each predicted a small amount of variance in transformational leadership. Passive/avoidant leadership was inversely predicted by the understanding emotions branch of EI. Overall, EI was not found to be a useful predictor of leadership style and leadership outcomes in the Main Study of this project. Implications for researchers and human resource practitioners are discussed.

Applying social technology to Business Process Lifecycle Management

In recent years social technologies such as wikis, blogs or microblogging have seen an exponential growth in the uptake of their user base making this type of technology one of the most significant networking and knowledge sharing platforms for potentially hundreds of millions of users. However, the adoption of these technologies has been so far mostly for private purposes. First attempts have been made to embed features of social technologies in the corporate IT landscape, and Business Process Management is no exception. This paper aims to consolidate the opportunities for integrating social technologies into the different stages of the business process lifecycle. Thus, it contributes to a conceptualization of this fast growing domain, and can help to categorize academic and corporate development activities.

Fitness and fatness in childhood obesity : implications of physical activity

Overweight and obesity are a significant cause of poor health worldwide, particularly in conjunction with low levels of physical activity (PA). PA is health-protective and essential for the physical growth and development of children, promoting physical and psychological health while simultaneously increasing the probability of remaining active as an adult. However, many obese children and adolescents have a unique set of physiological, biomechanical, and neuromuscular barriers to PA that they must overcome. It is essential to understand the influence of these barriers on an obese child's motivation in order to exercise and tailor exercise programs to the special needs of this population. Chapter Outline • Introduction • Defining Physical Activity, Exercise, and Physical Fitness • Physical Activity, Physical Fitness, And Motor Competence In Obese Children • Physical Activity and Obesity in Children • Physical Fitness in Obese Children • Balance and Gait in Obese Children • Motor Competence in Obese Children • Physical Activity Guidelines for Obese Children • Clinical Assessment of the Obese Child • Physical Activity Characteristics: Mode • Physical Activity Characteristics: Intensity • Physical Activity Characteristics: Frequency • Physical Activity Characteristics: Duration • Conclusion

Consumers’ intentions to adopt 3G in regional cities in China : a comparative study

Rapid mobile technological evolution and the large economic stake in commercial development of mobile technological innovation make it necessary to understand consumers' motivations towards the latest advanced and updated technologies and services. 3G (the third generation of mobile communication technology) recently started its commercial development in the world‘s largest mobile communication market, China, after being delayed for a few years. Although China fell behind in commercially developing 3G, it is difficult to ignore studying this area, given the size of the market and promising future developments. This market deserves focused research attention, especially in terms of consumer behaviour towards the adoption of mobile technological innovation. Thus, the program of research in this thesis was designed to investigate how Chinese consumers respond to the use of this newly launched mobile technological innovation, with a focus on what factors affect their 3G adoption intentions. It aimed to yield important insights into Chinese consumers‘ innovation adoption behaviours and to contribute to marketing and innovation adoption research. Furthermore, it has been documented that Chinese consumers vary widely between regions in dialect, lifestyle, culture, purchasing power and consumption attitudes. Based on economic development and local culture, China can be divided geographically into distinctive regional consumer markets. Consequently, the results of consumer behaviour research in one region may not necessarily be extrapolated to other regions. In order to better understand Chinese consumers, the disparities between regions should not be overlooked. Therefore, another objective of this program of research was to examine regional variances in consumers' innovation adoption, specifically to identify the similarities and differences in factors influencing 3G adoption, contributing to intra-cultural studies. An extensive literature review identified two gaps: current China-based innovation adoption research studies are limited in providing adequate prediction and explanation of Chinese consumers' intentions to adopt 3G; and there was limited knowledge about the differences between regional Chinese consumers in innovation adoption. Two research questions therefore were developed to address these gaps: 1) What factors influence Chinese consumers' intentions to adopt 3G? 2) How do Chinese consumers differ between regional markets in the relative influence of the factors in determining their intentions to adopt 3G? In accordance with postpositivist research philosophy, two studies were designed to answer the research questions, using mixed methods. To meet the research objectives, the two studies were both conducted in three regional cities, namely Beijing, Shanghai and Wuhan, centred in the three regions of North China, East China and Central China respectively, with sufficient cultural and economical regional variances. Study One was an exploratory study with qualitative research methods. It involved 45 in-depth interviews in the three research cities to gain rich insights into the research context from natural settings. Eight important concepts related to 3G adoption were generated from analysis of the interview data, namely utilitarian expectation, hedonic expectation, status gains, status loss avoidance, normative influence, external influence, cost and quality concern. The concepts of social loss avoidance and quality concern were two unique findings, whereas the other concepts were similar to the findings in Western innovation adoption studies. Moreover, variances in 3G adoption between three groups of regional consumers were also identified, focusing on the perceptions of two concepts, namely status gains and normative influence. The conceptual research model was then developed incorporating the eight concepts plus the dependent variable of adoption intention. The hypothesized relationships between the nine constructs and hypotheses about the differences between regional consumers in 3G adoption were informed by the findings of Study One and the literature reviewed. Study Two was a quantitative study involving a web-based survey and statistical analysis procedure. The web-based survey attracted 800 residents from the three research cities, 270 from Beijing, 265 from Shanghai and 265 from Wuhan. They comprised three research samples for this study and consequently three sets of data were obtained. The data was analysed by Structural Equation Modelling together with Multi-group Analysis. The analysis confirmed that the concepts generated in Study One were influential factors affecting Chinese consumers' 3G adoption intention, with the exception of the concept external influence. Differences were found between the samples in the three research cities in the effect of hedonic expectation, status gains, status loss avoidance and normative influence on 3G adoption intention. The two Studies undertaken in this thesis contributed a better understanding of Chinese consumers' intentions to adopt advanced mobile technological innovation, namely 3G, in three regional markets. This knowledge contributes to innovation adoption and intra-cultural research, as well as consumer behaviour theory. It is also able to inform international and domestic telecommunication companies to develop and deliver more effective marketing strategies across Chinese regional markets. Limitations in the research were identified in terms of the sampling techniques used and the design of the two Studies. Future research was suggested in other Chinese regional markets and into consumer adoption of other types of mobile technological innovations.

Die Geographie der Kreativen Klasse in Deutschland [The geography of the Creative Class in Germany]

We analyze the regional distribution of different categories of creative individuals in Germany. Generally, the share of creative people is higher in cities as compared to the rural area The freelancing artists are a kind of exception in this respect; they constitute a relatively high share of the population in some rural area A high share of creative people in a region can be explained by a high level of public provisions and a high share of foreign born population, which can be regarded as an indicator of the “openness” in the local milieu. Good employment opportunities have only a relatively weak impact. Regions with a high share of creatives tend to have an above average level of new business formation, a high level of innovation and a relatively high share of employees in high-tech industries.

Measurement of total body water (TBW) and total energy expenditure (TEE) using stable isotopes

Understanding the relationship between diet, physical activity and health in humans requires accurate measurement of body composition and daily energy expenditure. Stable isotopes provide a means of measuring total body water and daily energy expenditure under free-living conditions. While the use of isotope ratio mass spectrometry (IRMS) for the analysis of 2H (Deuterium) and 18O (Oxygen-18) is well established in the field of human energy metabolism research, numerous questions remain regarding the factors which influence analytical and measurement error using this methodology. This thesis was comprised of four studies with the following emphases. The aim of Study 1 was to determine the analytical and measurement error of the IRMS with regard to sample handling under certain conditions. Study 2 involved the comparison of TEE (Total daily energy expenditure) using two commonly employed equations. Further, saliva and urine samples, collected at different times, were used to determine if clinically significant differences would occur. Study 3 was undertaken to determine the appropriate collection times for TBW estimates and derived body composition values. Finally, Study 4, a single case study to investigate if TEE measures are affected when the human condition changes due to altered exercise and water intake. The aim of Study 1 was to validate laboratory approaches to measure isotopic enrichment to ensure accurate (to international standards), precise (reproducibility of three replicate samples) and linear (isotope ratio was constant over the expected concentration range) results. This established the machine variability for the IRMS equipment in use at Queensland University for both TBW and TEE. Using either 0.4mL or 0.5mL sample volumes for both oxygen-18 and deuterium were statistically acceptable (p>0.05) and showed a within analytical variance of 5.8 Delta VSOW units for deuterium, 0.41 Delta VSOW units for oxygen-18. This variance was used as “within analytical noise” to determine sample deviations. It was also found that there was no influence of equilibration time on oxygen-18 or deuterium values when comparing the minimum (oxygen-18: 24hr; deuterium: 3 days) and maximum (oxygen-18: and deuterium: 14 days) equilibration times. With regard to preparation using the vacuum line, any order of preparation is suitable as the TEE values fall within 8% of each other regardless of preparation order. An 8% variation is acceptable for the TEE values due to biological and technical errors (Schoeller, 1988). However, for the automated line, deuterium must be assessed first followed by oxygen-18 as the automated machine line does not evacuate tubes but merely refills them with an injection of gas for a predetermined time. Any fractionation (which may occur for both isotopes), would cause a slight elevation in the values and hence a lower TEE. The purpose of the second and third study was to investigate the use of IRMS to measure the TEE and TBW of and to validate the current IRMS practices in use with regard to sample collection times of urine and saliva, the use of two TEE equations from different research centers and the body composition values derived from these TEE and TBW values. Following the collection of a fasting baseline urine and saliva sample, 10 people (8 women, 2 men) were dosed with a doubly labeled water does comprised of 1.25g 10% oxygen-18 and 0.1 g 100% deuterium/kg body weight. The samples were collected hourly for 12 hrs on the first day and then morning, midday, and evening samples were collected for the next 14 days. The samples were analyzed using an isotope ratio mass spectrometer. For the TBW, time to equilibration was determined using three commonly employed data analysis approaches. Isotopic equilibration was reached in 90% of the sample by hour 6, and in 100% of the sample by hour 7. With regard to the TBW estimations, the optimal time for urine collection was found to be between hours 4 and 10 as to where there was no significant difference between values. In contrast, statistically significant differences in TBW estimations were found between hours 1-3 and from 11-12 when compared with hours 4-10. Most of the individuals in this study were in equilibrium after 7 hours. The TEE equations of Prof Dale Scholler (Chicago, USA, IAEA) and Prof K.Westerterp were compared with that of Prof. Andrew Coward (Dunn Nutrition Centre). When comparing values derived from samples collected in the morning and evening there was no effect of time or equation on resulting TEE values. The fourth study was a pilot study (n=1) to test the variability in TEE as a result of manipulations in fluid consumption and level of physical activity; the magnitude of change which may be expected in a sedentary adult. Physical activity levels were manipulated by increasing the number of steps per day to mimic the increases that may result when a sedentary individual commences an activity program. The study was comprised of three sub-studies completed on the same individual over a period of 8 months. There were no significant changes in TBW across all studies, even though the elimination rates changed with the supplemented water intake and additional physical activity. The extra activity may not have sufficiently strenuous enough and the water intake high enough to cause a significant change in the TBW and hence the CO2 production and TEE values. The TEE values measured show good agreement based on the estimated values calculated on an RMR of 1455 kcal/day, a DIT of 10% of TEE and activity based on measured steps. The covariance values tracked when plotting the residuals were found to be representative of “well-behaved” data and are indicative of the analytical accuracy. The ratio and product plots were found to reflect the water turnover and CO2 production and thus could, with further investigation, be employed to identify the changes in physical activity.

Exploiting the 1,2,3-triazolium motif in anion-templated formation of a bromide-selective rotaxane host assembly

Stimulated by the efficacy of copper (I) catalysed Huisgen-type 1,3-dipolar cycloaddition of terminal alkynes and organic azides to generate 1,4-disubstituted 1,2,3-triazole derivatives, the importance of ‘click’ chemistry in the synthesis of organic and biological molecular systems is ever increasing.[1] The mild reaction conditions have also led to this reaction gaining favour in the construction of interlocked molecular architectures.[2-4] In the majority of cases however, the triazole group simply serves as a covalent linkage with no function in the resulting organic molecular framework. More recently a renewed interest has been shown in the transition metal coordination chemistry of triazole ligands.[3, 5, 6] In addition novel aryl macrocyclic and acyclic triazole based oligomers have been shown to recognise halide anions via cooperative triazole C5-H….anion hydrogen bonds.[7] In light of this it is surprising the potential anion binding affinity of the positively charged triazolium motif has not, with one notable exception,[8] been investigated. With the objective of manipulating the unique topological cavities of mechanically bonded molecules for anion recognition purposes, we have developed general methods of using anions to template the formation of interpenetrated and interlocked structures.[9-13] Herein we report the first examples of exploiting the 1,2,3-triazolium group in the anion templated formation of pseudorotaxane and rotaxane assemblies. In an unprecedented discovery the bromide anion is shown to be a superior templating reagent to chloride in the synthesis of a novel triazolium axle containing [2]rotaxane. Furthermore the resulting rotaxane interlocked host system exhibits the rare selectivity preference for bromide over chloride...

SwarmPS: Rapid, semi-automated single particle selection software

Single particle analysis (SPA) coupled with high-resolution electron cryo-microscopy is emerging as a powerful technique for the structure determination of membrane protein complexes and soluble macromolecular assemblies. Current estimates suggest that ∼104–105 particle projections are required to attain a 3 Å resolution 3D reconstruction (symmetry dependent). Selecting this number of molecular projections differing in size, shape and symmetry is a rate-limiting step for the automation of 3D image reconstruction. Here, we present SwarmPS, a feature rich GUI based software package to manage large scale, semi-automated particle picking projects. The software provides cross-correlation and edge-detection algorithms. Algorithm-specific parameters are transparently and automatically determined through user interaction with the image, rather than by trial and error. Other features include multiple image handling (∼102), local and global particle selection options, interactive image freezing, automatic particle centering, and full manual override to correct false positives and negatives. SwarmPS is user friendly, flexible, extensible, fast, and capable of exporting boxed out projection images, or particle coordinates, compatible with downstream image processing suites.

The beliefs which motivate young male and female drivers to speed : a comparison of low and high intenders

In Australia, young drivers aged 17 to 24 years, and particularly males, have the highest risk of being involved in a fatal crash. Investigation of young drivers’ beliefs allows for a greater understanding of their involvement in risky behaviours, such as speeding, as beliefs are associated with intentions, the antecedent to behaviour. The theory of planned behaviour (TPB) was used to conceptualise beliefs using a scenario based questionnaire distributed to licensed drivers (N = 398). The questionnaire measured individual’s beliefs and intentions to speed in a particular situation. Consistent with a TPB-based approach, the beliefs of those with low intentions to speed (‘low intenders’) were compared with the beliefs of those with high intentions (‘high intenders’) with such comparisons conducted separately for males and females. Overall, significant differences in the beliefs held by low and high intenders and for both females and males were found. Specifically, for females, it was found that high intenders were significantly more likely to perceive advantages of speeding, less likely to perceive disadvantages, and more likely to be encouraged to speed on familiar and inappropriately signed roads than female low intenders. Females, however, did not differ in their perceptions of support from friends, with all females reporting some level of disapproval from most friends and all females (i.e., low and high intenders) reporting approval to speed from their male friends. The results for males revealed that high intenders were significantly more likely to speed on familiar and inappropriately signed roads as well as having greater perceptions of support from all friends, except from those friends with whom they worked. Low and high intending males did not differ in their perceptions of the advantages and disadvantages of speeding, with the exception of feelings of excitement whereby high intenders reported speeding to be more exciting than low intenders. The findings are discussed in terms of how they may directly inform the content of mass media and public education campaigns aimed at encouraging young drivers to slow down.

Qualified indefeasibility and the careless mortgagee

In recent years a great deal of case law has been generated in relation to mortgages where the mortgagee has not engaged in adequate identity verification of the mortgagor and the mortgage has subsequently been found to be forged. As a result, careless mortgagee provisions operate in Queensland as an exception to indefeasibility. Similar provisions are expected to commence soon in New South Wales. This article examines the mortgagee’s position with the benefit of indefeasibility and then considers the impact of the careless mortgagee provisions on the rights of a mortgagee under a forged mortgage, concluding that the provisions significantly change the dynamic between a registered mortgagee and registered owner who has not signed the mortgage. These provisions appear to give the mortgagee a conditional indefeasibility, with the intention of reducing the State’s exposure to the payment of compensation in the case of identity fraud. They are however, more successful in the case of forgery by a third party rather than forgery by a co-owner.

Towards the biofortification of banana fruit for enhanced micronutrient content

In Uganda, vitamin A deficiency (VAD) and iron deficiency anaemia (IDA) are major public health problems with between 15-32% of children under 5 years of age showing VAD and 73% being anaemic. This is largely due to the fact that the staple food crop of the country, banana, is low in pro-vitamin A and iron, therefore leading to dietary deficiencies. Although worldwide progress has been made to control VAD and IDA through supplementation, food fortification and diet diversification, their long term sustainability and impact in developing countries such as Uganda is limited. The approach taken by researchers at Queensland University of Technology (QUT), Australia, in collaboration with the National Agricultural Research Organization (NARO), Uganda, to address this problem, is to generate consumer acceptable banana varieties with significantly increased levels of pro-vitamin A and iron in the fruit using genetic engineering techniques. Such an approach requires the use of suitable, well characterised genes and promoters for targeted transgene expression. Recently, a new banana phytoene synthase gene (APsy2a) involved in the synthesis of pro-vitamin A (pVA) carotenoids was isolated from a high â-carotene banana (F’ei cv Asupina). In addition, sequences of banana ferritin, an iron storage protein, have been isolated from Cavendish banana. The aim of the research described in this thesis was to evaluate the function of these genes to assess their suitability for the biofortification of banana fruit. In addition, a range of banana-derived promoters were characterised to determine their suitability for controlling the expression of transgenes in banana fruit. Due to the time constraints involved with generating transgenic banana fruit, rice was used as the model crop to investigate the functionality of the banana-derived APsy2a and ferritin genes. Using Agrobacterium-mediated transformation, rice callus was transformed with APsy2a +/- the bacterial-derived carotene desaturase gene (CrtI) each under the control of the constitutive maize poly-ubiquitin promoter (ZmUbi) or seed-specific rice glutelin1 (Gt1) promoter. The maize phytoene synthase (ZmPsy1) gene was included as a control. On selective media, with the exception of ZmUbi-CrtI-transgenic callus, all antibiotic resistant callus displayed a yellow-orange colour from which the presence of â-carotene was demonstrated using Raman spectroscopy. Although the regeneration of plants from yellow-orange callus was difficult, 16 transgenic plants were obtained and characterised from callus transformed with ZmUbi-APys2a alone. At least 50% of the T1 seeds developed a yellow-orange coloured callus which was found to contain levels of â-carotene ranging from 4.6-fold to 72-fold higher than that in non-transgenic rice callus. Using the seed-specific Gt1 promoter, 38 transgenic rice plants were generated from APsy2a-CrtI-transformed callus while 32 plants were regenerated from ZmPsy1-CrtI-transformed callus. However, when analysed for presence of transgene by PCR, all transgenic plants contained the APsy2a, ZmPsy1 or CrtI transgene, with none of the plants found to be co-transformed. Using Raman spectroscopy, no â-carotene was detected in-situ in representative T1 seeds. To investigate the potential of the banana-derived ferritin gene (BanFer1) to enhance iron content, rice callus was transformed with constitutively expressed BanFer1 using the soybean ferritin gene (SoyFer) as a control. A total of 12 and 11 callus lines independently transformed with BanFer1 and SoyFer, respectively, were multiplied and transgene expression was verified by RT-PCR. Pearl’s Prussian blue staining for in-situ detection of ferric iron showed a stronger blue colour in rice callus transformed with BanFer1 compared to SoyFer. Using flame atomic absorption spectrometry, the highest mean amount of iron quantified in callus transformed with BanFer1 was 30-fold while that obtained using the SoyFer was 14-fold higher than the controls. In addition, ~78% of BanFer1-transgenic callus lines and ~27% of SoyFer-transgenic callus lines had significantly higher iron content than the non-transformed controls. Since the genes used for enhancing micronutrient content need to be expressed in banana fruit, the activity of a range of banana-derived, potentially fruit-active promoters in banana was investigated. Using uidA (GUS) as a reporter gene, the function of the Expansin1 (MaExp1), Expansin1 containing the rice actin intron (MaExp1a), Expansin4 (MaExp4), Extensin (MaExt), ACS (MaACS), ACO (MaACO), Metallothionein (MaMT2a) and phytoene synthase (APsy2a) promoters were transiently analysed in intact banana fruit using two transformation methods, particle bombardment and Agrobacterium-mediated infiltration (agro-infiltration). Although a considerable amount of variation in promoter activity was observed both within and between experiments, similar trends were obtained using both transformation methods. The MaExp1 and MaExp1a directed high levels of GUS expression in banana fruit which were comparable to those observed from the ZmUbi and Banana bunchy top virus-derived BT4 promoters that were included as positive controls. Lower levels of promoter activity were obtained in both methods using the MaACO and MaExt promoters while the MaExp4, MaACS, and APsy2a promoters directed the lowest GUS activity in banana fruit. An attempt was subsequently made to use agro-infiltration to assess the expression of pVA biosynthesis genes in banana fruit by infiltrating fruit with constructs in which the ZmUbi promoter controlled the expression of APsy2a +/- CrtI, and with the maize phytoene synthase gene (ZmPsy1) included as a control. Unfortunately, the large amount of variation and inconsistency observed within and between experiments precluded any meaningful conclusions to be drawn. The final component of this research was to assess the level of promoter activity and specificity in non-target tissue. These analyses were done on leaves obtained from glasshouse-grown banana plants stably transformed with MaExp1, MaACO, APsy2a, BT4 and ZmUbi promoters driving the expression of the GUS gene in addition to leaves from a selection of the same transgenic plants which were growing in a field trial in North Queensland. The results from both histochemical and fluorometric GUS assays showed that the MaExp1 and MaACO promoters directed very low GUS activities in leaves of stably transformed banana plants compared to the constitutive ZmUbi and BT4 promoters. In summary, the results from this research provide evidence that the banana phytoene synthase gene (APsy2a) and the banana ferritin gene (BanFer1) are functional, since the constitutive over-expression of each of these transgenes led to increased levels of pVA carotenoids (for APsy2a) and iron content (for BanFer1) in transgenic rice callus. Further work is now required to determine the functionality of these genes in stably-transformed banana fruit. This research also demonstrated that the MaExp1 and MaACO promoters are fruit-active but have low activity in non-target tissue (leaves), characteristics that make them potentially useful for the biofortification of banana fruit. Ultimately, however, analysis of fruit from field-grown transgenic plants will be required to fully evaluate the suitability of pVA biosynthesis genes and the fruit-active promoters for fruit biofortification.