Optimization of human papillomavirus type 16 (HPV-16) L1 expression in plants: Comparison of the suitability of different HPV-16 L1 gene variants and different cell-compartment localization

Virus-like particle-based vaccines for high-risk human papillomaviruses (HPVs) appear to have great promise; however, cell culture-derived vaccines will probably be very expensive. The optimization of expression of different codon-optimized versions of the HPV-16 L1 capsid protein gene in plants has been explored by means of transient expression from a novel suite of Agrobacterium tumefaciens binary expression vectors, which allow targeting of recombinant protein to the cytoplasm, endoplasmic reticulum (ER) or chloroplasts. A gene resynthesized to reflect human codon usage expresses better than the native gene, which expresses better than a plant-optimized gene. Moreover, chloroplast localization allows significantly higher levels of accumulation of L1 protein than does cytoplasmic localization, whilst ER retention was least successful. High levels of L1 (>17% total soluble protein) could be produced via transient expression: the protein assembled into higher-order structures visible by electron microscopy, and a concentrated extract was highly immunogenic in mice after subcutaneous injection and elicited high-titre neutralizing antibodies. Transgenic tobacco plants expressing a human codon-optimized gene linked to a chloroplast-targeting signal expressed L1 at levels up to 11% of the total soluble protein. These are the highest levels of HPV L1 expression reported for plants: these results, and the excellent immunogenicity of the product, significantly improve the prospects of making a conventional HPV vaccine by this means. © 2007 SGM.

Improved molecular tools for sugar cane biotechnology

Sugar cane is a major source of food and fuel worldwide. Biotechnology has the potential to improve economically-important traits in sugar cane as well as diversify sugar cane beyond traditional applications such as sucrose production. High levels of transgene expression are key to the success of improving crops through biotechnology. Here we describe new molecular tools that both expand and improve gene expression capabilities in sugar cane. We have identified promoters that can be used to drive high levels of gene expression in the leaf and stem of transgenic sugar cane. One of these promoters, derived from the Cestrum yellow leaf curling virus, drives levels of constitutive transgene expression that are significantly higher than those achieved by the historical benchmark maize polyubiquitin-1 (Zm-Ubi1) promoter. A second promoter, the maize phosphonenolpyruvate carboxylate promoter, was found to be a strong, leaf-preferred promoter that enables levels of expression comparable to Zm-Ubi1 in this organ. Transgene expression was increased approximately 50-fold by gene modification, which included optimising the codon usage of the coding sequence to better suit sugar cane. We also describe a novel dual transcriptional enhancer that increased gene expression from different promoters, boosting expression from Zm-Ubi1 over eightfold. These molecular tools will be extremely valuable for the improvement of sugar cane through biotechnology.

The complete mitochondrial genome of the yellow coaster, Acraea issoria (Lepidoptera: Nymphalidae: Heliconiinae: Acraeini): sequence, gene organization and a unique tRNA translocation event

In this paper, the complete mitochondrial genome of Acraea issoria (Lepidoptera: Nymphalidae: Heliconiinae: Acraeini) is reported; a circular molecule of 15,245 bp in size. For A. issoria, genes are arranged in the same order and orientation as the complete sequenced mitochondrial genomes of the other lepidopteran species, except for the presence of an extra copy of tRNAIle(AUR)b in the control region. All protein-coding genes of A. issoria mitogenome start with a typical ATN codon and terminate in the common stop codon TAA, except that COI gene uses TTG as its initial codon and terminates in a single T residue. All tRNA genes possess the typical clover leaf secondary structure except for tRNASer(AGN), which has a simple loop with the absence of the DHU stem. The sequence, organization and other features including nucleotide composition and codon usage of this mitochondrial genome were also reported and compared with those of other sequenced lepidopterans mitochondrial genomes. There are some short microsatellite-like repeat regions (e.g., (TA)9, polyA and polyT) scattered in the control region, however, the conspicuous macro-repeats units commonly found in other insect species are absent.

Management and organisational barriers in the acquisition of computer usage skills by mature age workers

Aim: To investigate workplace cultures in the acquisition of computer usage skills by mature age workers. Methods: Data were gathered through focus groups conducted at job network centres in the Greater Brisbane metropolitan region. Participants who took part were a mixture of workers and job-seekers. Results: The results suggest that mature age workers can be exposed to inappropriate computer training practices and age-insensitive attitudes towards those with low base computer skills. Conclusions: There is a need for managers to be observant of ageist attitudes in the work place and to develop age-sensitive strategies to help mature age workers learn computer usage skills. Mature age workers also need to develop skills in ways which are practical and meaningful to their work.

Determinants of pre-hospital care non-usage for patients with emergency care needs [including commentary by Snooks H.]

Objective-To establish the demographic, health status and insurance determinants of pre-hospital ambulance non-usage for patients with emergency medical needs. Methods-Triage category, date of birth, sex, marital status, country of origin, method and time of arrival, ambulance insurance status, diagnosis, and disposal were collected for all patients who presented over a four month period (n=10 229) to the emergency department of a major provincial hospital. Data for patients with urgent (n=678) or critical care needs (n=332) who did not use pre-hospital care were analysed using Poisson regression. Results-Only a small percentage (6.6%) of the total sample were triaged as having urgent medical needs or critical care needs (3.2%). Predictors of usage for those with urgent care needs included age greater than 65 years (prevalence ratio (PR)=0.54; 95% confidence interval (CI)= 0.35 to 0.83), being admitted to intensive care or transferred to another hospital (PR=0.62; 95% CI=0.44 to 0.89) or ward (PR=0.72; 95% CI=0.56 to 0.93) and ambulance insurance status (PR=0.67; 95% CI=052 to 0.86). Sex, marital status, time of day and country of origin were not predictive of usage and non-usage. Predictors of usage for those with critical care needs included age 65 years or greater (PR=0.45; 95% CI=0.25 to 0.81) and a diagnosis of trauma (PR=0.49; 95% CI=0.26 to 0.92). A non-English speaking background was predictive of non-usage (PR=1.98; 95% CI=1.06 to 3.70). Sex, marital status, time of day, triage and ambulance insurance status were not predictive of non-usage. Conclusions-Socioeconomic and medical factors variously influence ambulance usage depending on the severity or urgency of the medical condition. Ambulance insurance status was less of an influence as severity of condition increased suggesting that, at a critical level of urgency, patients without insurance are willing to pay for a pre-hospital ambulance service.

A visualisation framework for collaborative virtual environment usage information

Due to the popularity of modern Collaborative Virtual Environments, there has been a related increase in their size and complexity. Developers therefore need visualisations that expose usage patterns from logged data, to understand the structures and dynamics of these complex environments. This chapter presents a new framework for the process of visualising virtual environment usage data. Major components, such as an event model, designer task model and data acquisition infrastructure are described. Interface and implementation factors are also developed, along with example visualisation techniques that make use of the new task and event model. A case study is performed to illustrate a typical scenario for the framework, and its benefits to the environment development team.

Key performance indicators for innovation implementation : perception vs. actual usage

This paper aimed to explore the proportion associated with the perceived importance and the actual use of performance indicators from manufacturing and non manufacturing industries. The sample was 86 small and medium sized-organizations in Thailand. The perceived importance and the actual use of financial and non financial indicators were found to be significantly related among manufacturing and non manufacturing industries. KPI 3, 9, and 12 (i.e. sales and sales growth; quality of products and /or services; and process time) were perceived the most importance among manufacturing managers (85.3%, 79.4% and 76.5% respectively). While KPI 6, 9, and 12 (i.e. customer satisfaction, quality of products and /or services; and process time) were perceived the most importance among non manufacturing managers (84.8%, 93.5%, and 84.8% respectively). Interestingly, the most used KPIs for manufacturing were sales and sales growth (64.7%); profit margins (61.8%); and customer satisfaction (84.8) while non manufacturing used quality products/services (60.9%); sales and sales growth (54.3%) and employee development (54.3%) respectively. Limitation and implication were also discussed.

Functional outcome and usage of the prosthesis of lower limb amputees fitted with osseointegrated fixation

This study established that the core principle underlying categorisation of activities have the potential to provide more comprehensive outcomes than the recognition of activities because it takes into consideration activities other than directional locomotion.

On the usage of labels and icons in business process modeling

The value of business process models is dependent not only on the choice of graphical elements in the model, but also on their annotation with additional textual and graphical information. This research discusses the use of text and icons for labeling the graphical constructs in a process model. We use two established verb classification schemes to examine the choice of activity labels in process modeling practice. Based on our findings, we synthesize a set of twenty-five activity label categories. We propose a systematic approach for graphically representing these label categories through the use of graphical icons, such that the resulting process models are easier and more readily understandable by end users. Our findings contribute to an ongoing stream of research investigating the practice of process modeling and thereby contribute to the body of knowledge about conceptual modeling quality overall.