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.

Sound media musicianship

By the end of the 20th century the shift from professional recording studio to personal computer based recording systems was well established (Chadabe 1997) and musicians could increasingly see the benefits of value adding to the musical process by producing their own musical endeavours. At the Queensland University of Technology (QUT) where we were teaching, the need for a musicianship program that took account of these trends was becoming clear. The Sound Media Musicianship unit described in this chapter was developed to fill this need and ran from 1999 through 2010.

A survey of tagging techniques for music, speech and environmental sound

Sound tagging has been studied for years. Among all sound types, music, speech, and environmental sound are three hottest research areas. This survey aims to provide an overview about the state-of-the-art development in these areas.We discuss about the meaning of tagging in different sound areas at the beginning of the journey. Some examples of sound tagging applications are introduced in order to illustrate the significance of this research. Typical tagging techniques include manual, automatic, and semi-automatic approaches.After reviewing work in music, speech and environmental sound tagging, we compare them and state the research progress to date. Research gaps are identified for each research area and the common features and discriminations between three areas are discovered as well. Published datasets, tools used by researchers, and evaluation measures frequently applied in the analysis are listed. In the end, we summarise the worldwide distribution of countries dedicated to sound tagging research for years.

Towards persistent indoor appearance-based localization, mapping and navigation using CAT-Graph

The challenge of persistent appearance-based navigation and mapping is to develop an autonomous robotic vision system that can simultaneously localize, map and navigate over the lifetime of the robot. However, the computation time and memory requirements of current appearance-based methods typically scale not only with the size of the environment but also with the operation time of the platform; also, repeated revisits to locations will develop multiple competing representations which reduce recall performance. In this paper we present a solution to the persistent localization, mapping and global path planning problem in the context of a delivery robot in an office environment over a one-week period. Using a graphical appearance-based SLAM algorithm, CAT-Graph, we demonstrate constant time and memory loop closure detection with minimal degradation during repeated revisits to locations, along with topological path planning that improves over time without using a global metric representation. We compare the localization performance of CAT-Graph to openFABMAP, an appearance-only SLAM algorithm, and the path planning performance to occupancy-grid based metric SLAM. We discuss the limitations of the algorithm with regard to environment change over time and illustrate how the topological graph representation can be coupled with local movement behaviors for persistent autonomous robot navigation.

Energy-efficient localization : GPS duty cycling with radio ranging

GPS is a commonly used and convenient technology for determining absolute position in outdoor environments, but its high power consumption leads to rapid battery depletion in mobile devices. An obvious solution is to duty cycle the GPS module, which prolongs the device lifetime at the cost of increased position uncertainty while the GPS is off. This article addresses the trade-off between energy consumption and localization performance in a mobile sensor network application. The focus is on augmenting GPS location with more energy-efficient location sensors to bound position estimate uncertainty while GPS is off. Empirical GPS and radio contact data from a large-scale animal tracking deployment is used to model node mobility, radio performance, and GPS. Because GPS takes a considerable, and variable, time after powering up before it delivers a good position measurement, we model the GPS behaviour through empirical measurements of two GPS modules. These models are then used to explore duty cycling strategies for maintaining position uncertainty within specified bounds. We then explore the benefits of using short-range radio contact logging alongside GPS as an energy-inexpensive means of lowering uncertainty while the GPS is off, and we propose strategies that use RSSI ranging and GPS back-offs to further reduce energy consumption. Results show that our combined strategies can cut node energy consumption by one third while still meeting application-specific positioning criteria.

Long exposure localization in darkness using consumer cameras

In this paper we demonstrate passive vision-based localization in environments more than two orders of magnitude darker than the current benchmark using a 100 webcam and a 500 camera. Our approach uses the camera’s maximum exposure duration and sensor gain to achieve appropriately exposed images even in unlit night-time environments, albeit with extreme levels of motion blur. Using the SeqSLAM algorithm, we first evaluate the effect of variable motion blur caused by simulated exposures of 132 ms to 10000 ms duration on localization performance. We then use actual long exposure camera datasets to demonstrate day-night localization in two different environments. Finally we perform a statistical analysis that compares the baseline performance of matching unprocessed greyscale images to using patch normalization and local neighbourhood normalization – the two key SeqSLAM components. Our results and analysis show for the first time why the SeqSLAM algorithm is effective, and demonstrate the potential for cheap camera-based localization systems that function across extreme perceptual change.

Load bearing improves the limits of agreement for repeated in vivo measures of the speed of sound in human Achilles tendon

Axial acoustic wave propagation has been widely used in evaluating the mechanical properties of human bone in vivo. However, application of this technique to monitor soft tissues, such as tendon, has received comparatively little scientific attention. Laboratory-based research has established that axial acoustic wave transmission is not only related to the physical properties of equine tendon but is also proportional to tensile load to which it is exposed (Miles et al., 1996; Pourcelot et al., 2005). The reproducibility of the technique for in vivo measurements in human tendon, however, has not been established. The aim of this study was to evaluate the limits of agreement for repeated measures of the speed of sound (SoS) in human Achilles tendon in vivo. Methods: A custom built ultrasound device, consisting of an A-mode 1MHz emitter and two regularly spaced receivers, was used to measure the SoS in the mid-portion of the Achilles tendon in ten healthy males and ten females (mean age: 33.8 years, range 23-56 yrs; height: 1.73±0.08 m; weight: 68.4±15.3 kg). The emitter and receivers were held at fixed positions by a polyethylene frame and maintained in close contact with the skin overlying the tendon by means of elasticated straps. Repeated SoS measurements were taken with the subject prone (non-weightbearing and relaxed Achilles tendon) and during quiet bipedal and unipedal stance. In each instance, the device was detached and repositioned prior to measurement. Results: Limits of agreement for repeated SoS measures during non-weightbearing and bipedal and unipedal stance were ±53, ±28 and ±21 m/s, respectively. The average SoS in the non-weightbearing Achilles tendon was 1804±198 m/s. There was a significant increase in the average SoS during bilateral (2122±135 m/s) (P < 0.05) and unilateral (2221±79 m/s) stance (P < 0.05). Conclusions: Repeated SoS measures in human Achilles tendon were more reliable during stance than under non-weightbearing conditions. These findings are consistent with previous research in equine tendon in which lower variability in SoS was observed with increasing tensile load (Crevier-Denoix et al, 2009). Since the limits of agreement for Achilles tendon SoS are nearly 5% of the changes previously observed during walking and therapeutic heel raise exercises, acoustic wave transmission provides a promising new non-invasive method for determining tendon properties during sports and rehabilitation related activities.

Tracing the influence of non-narrative film and expanded cinema sound on experimental music

This study surveys and interrogates key conceptual frameworks and artistic practises that flow through the distinct but interconnected traditions of non-narrative film and experimental music, and examines how these are articulated in my own creative sound practise.

Gradual training of cascaded shape regression for facial landmark localization and pose estimation

Facial landmarks play an important role in face recognition. They serve different steps of the recognition such as pose estimation, face alignment, and local feature extraction. Recently, cascaded shape regression has been proposed to accurately locate facial landmarks. A large number of weak regressors are cascaded in a sequence to fit face shapes to the correct landmark locations. In this paper, we propose to improve the method by applying gradual training. With this training, the regressors are not directly aimed to the true locations. The sequence instead is divided into successive parts each of which is aimed to intermediate targets between the initial and the true locations. We also investigate the incorporation of pose information in the cascaded model. The aim is to find out whether the model can be directly used to estimate head pose. Experiments on the Annotated Facial Landmarks in the Wild database have shown that the proposed method is able to improve the localization and give accurate estimates of pose.

Familial typical migraine: Significant linkage and localization of a gene to Xq24-28

In a previous study we found evidence for an X-linked genetic component for familial typical migraine in two large Australian white pedigrees, designated MF7 and MF14. Significant excess allele sharing was indicated by nonparametric linkage (NPL) analysis using GENEHUNTER (P=0.031 and P=0.012, respectively), with a combined analysis of the two pedigrees showing further increased evidence for linkage, producing a maximum NPL score of 2.87 (P=0.011 ) at DXS 1123 on Xq27. The present study was aimed at refining the localization of the migraine X-chromosomal component by typing additional markers, performing haplotype analysis and applying a more powerful technique in the analysis of linkage data from these two pedigrees. Results from the haplotype analyses, coupled with linkage analyses that produced a peak GENEHUNTER-PLUS LOD* score of 2.388 (P=0.0005), provide compelling evidence for the presence of a migraine susceptibility locus on chromosome Xq24-28.

Sound Musicianship : Understanding the Crafts of Music

Sound Musicianship is a book for music educators and musicians about musicianship—about musical skills, abilities, habits, sensibilities and understandings. Musicianship is explored as a form of craftsmanship. Like most crafts, music requires a balance of theoretical knowledge and practical skills that contribute to a highly tuned ability to appreciate and express music. In particular, the book explores general trends that influence musicianship in the twenty-first century, such as an increased reliance on digital media, greater awareness of the neurological basis for musical behaviour, a renewed interest in connections between bodily movements and musical expression, and increased cultural plurality resulting from more frequent travel, increased levels of migration and ubiquitous telecommunications. The book has a deliberate focus on the developmental aspects of musicianship, which will benefit those hoping to advance their own music learning or that of others. It includes a diverse range of views and perspectives on musicianship and is organised into five sections. The first four sections explore the implications of music understood as sound, experience, motion and culture, respectively. In these sections, leading researchers and thinkers outline important issues and debates that are relevant to developing the crafts of music making and they share insights into recent trends and understandings. The final section of the book looks at educational considerations and provides a series of case studies that document innovative approaches to developing musicianship. Readers will encounter some new, interesting and thought-provoking ideas within these pages. As we move further into the twenty-first century—with all the opportunities and challenges for music making it brings—the requirement to review our concepts of musicianship training will intensify, and the definition of a “sound basis” for a contemporary musicianship will evolve. This book is intended to help stimulate and inform that evolutionary process.

Soundage : a practice-led approach to Gertrude Stein, sound, and generative language

In this practice-led research project I work to show how a re-reading and a particular form of listening to the sound-riddled nature of Gertrude Stein's work, Two: Gertrude Stein and her Brother, presents us with a contemporary theory of sound in language. This theory, though in its infancy, is a particular enjambment of sounded language that presents itself as an event, engaged with meaning, with its own inherent voice. It displays a propensity through engagement with the 'other' to erupt into love. In this thesis these qualities are reverberated further through the work of Seth Kim-Cohen's notion of the non-cochlear, Simon Jarvis's notion of musical thinking, Jean-Jacques Lecercle's notion of délire or nonsense, Luce Irigaray's notion of jouissant love and the Bracha Ettinger's notion of the generative matrixial border space. This reading then is simultaneously paired with my own work of scoring and creating a digital opera from Stein's work, thereby testing and performing Stein's theory. In this I show how a re-reading and relistening to Stein's work can be significant to feminist ethical language frames, contemporary philosophy, sonic art theory and digital language frames. Further significance of this study is that when the reverberation of Stein's engagements with language through sound can be listened to, a pattern emerges, one that encouragingly problematizes subjectivity and interweaves genres/methods and means, creating a new frame for sound in language, one with its own voice that I call soundage.

Localization of Mineralocorticoid Receptors at Mammalian Synapses

In the brain, membrane associated nongenomic steroid receptors can induce fast-acting responses to ion conductance and second messenger systems of neurons. Emerging data suggest that membrane associated glucocorticoid and mineralocorticoid receptors may directly regulate synaptic excitability during times of stress when adrenal hormones are elevated. As the key neuron signaling interface, the synapse is involved in learning and memory, including traumatic memories during times of stress. The lateral amygdala is a key site for synaptic plasticity underlying conditioned fear, which can both trigger and be coincident with the stress response. A large body of electrophysiological data shows rapid regulation of neuronal excitability by steroid hormone receptors. Despite the importance of these receptors, to date, only the glucocorticoid receptor has been anatomically localized to the membrane. We investigated the subcellular sites of mineralocorticoid receptors in the lateral amygdala of the Sprague-Dawley rat. Immunoblot analysis revealed the presence of mineralocorticoid receptors in the amygdala. Using electron microscopy, we found mineralocorticoid receptors expressed at both nuclear including: glutamatergic and GABAergic neurons and extra nuclear sites including: presynaptic terminals, neuronal dendrites, and dendritic spines. Importantly we also observed mineralocorticoid receptors at postsynaptic membrane densities of excitatory synapses. These data provide direct anatomical evidence supporting the concept that, at some synapses, synaptic transmission is regulated by mineralocorticoid receptors. Thus part of the stress signaling response in the brain is a direct modulation of the synapse itself by adrenal steroids.

Quantitative proteomic approach to study subcellular localization of membrane proteins

As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-state distribution of hundreds of integral membrane proteins within organelles. The technique uses a partial membrane fractionation strategy in conjunction with quantitative proteomics. Localization of proteins is achieved by measuring their distribution pattern across the density gradient using amine-reactive isotope tagging and comparing these patterns with those of known organelle residents. LOPIT relies on the assumption that proteins belonging to the same organelle will co-fractionate. Multivariate statistical tools are then used to group proteins according to the similarities in their distributions, and hence localization without complete centrifugal separation is achieved. The protocol requires approximately 3 weeks to complete and can be applied in a high-throughput manner to material from many varied sources.

Sub-cellular localization of membrane proteins

In eukaryotes, numerous complex sub-cellular structures exist. The majority of these are delineated by membranes. Many proteins are trafficked to these in order to be able to carry out their correct physiological function. Assigning the sub-cellular location of a protein is of paramount importance to biologists in the elucidation of its role and in the refinement of knowledge of cellular processes by tracing certain activities to specific organelles. Membrane proteins are a key set of proteins as these form part of the boundary of the organelles and represent many important functions such as transporters, receptors, and trafficking. They are, however, some of the most challenging proteins to work with due to poor solubility, a wide concentration range within the cell and inaccessibility to many of the tools employed in proteomics studies. This review focuses on membrane proteins with particular emphasis on sub-cellular localization in terms of methodologies that can be used to determine the accurate location of membrane proteins to organelles. We also discuss what is known about the membrane protein cohorts of major organelles.