Whole-genome multiparametric screening to identify modulators of epithelial-to-mesenchymal transition

Metastasis accounts for the poor prognosis of the majority of solid tumors. The phenotypic transition of nonmotile epithelial tumor cells to migratory and invasive “mesenchymal” cells (epithelial-to-mesenchymal transition [EMT]) enables the transit of cancer cells from the primary tumor to distant sites. There is no single marker of EMT; rather, multiple measures are required to define cell state. Thus, the multiparametric capability of high-content screening is ideally suited for the comprehensive analysis of EMT regulators. The aim of this study was to generate a platform to systematically identify functional modulators of tumor cell plasticity using the bladder cancer cell line TSU-Pr1-B1 as a model system. A platform enabling the quantification of key EMT characteristics, cell morphology and mesenchymal intermediate filament vimentin, was developed using the fluorescent whole-cell-tracking reagent CMFDA and a fluorescent promoter reporter construct, respectively. The functional effect of genome-wide modulation of protein-coding genes and miRNAs coupled with those of a collection of small-molecule kinase inhibitors on EMT was assessed using the Target Activation Bioapplication integrated in the Cellomics ArrayScan platform. Data from each of the three screens were integrated to identify a cohort of targets that were subsequently examined in a validation assay using siRNA duplexes. Identification of established regulators of EMT supports the utility of this screening approach and indicated capacity to identify novel regulators of this plasticity program. Pathway analysis coupled with interrogation of cancer-related expression profile databases and other EMT-related screens provided key evidence to prioritize further experimental investigation into the molecular regulators of EMT in cancer cells.

Whole-of-Curriculum Design in Law

In this book teaching professionalism is characterised by the scholarly underpinning of each contribution; and every contribution provides a rich resource for enhancing teaching practice. The critical concerns for legal education have been identified and discussed: curriculum design that includes graduate attributes; embedding specific attributes across the curriculum; empowering students to learn; academic teamwork to manage large student cohorts; first year and final year transition strategies; tracking students' personal development through the use of ePortfolio; assessment strategies; improving student well-being and promoting resilience; teaching practice to achieve deep learning; flexibility in delivery; the use of Web 2.0 technology; and understanding the 21st century student.

Methods for extracting genomic DNA from whole blood samples : current perspectives

Deoxyribonucleic acid (DNA) extraction has considerably evolved since it was initially performed back in 1869. It is the first step required for many of the available downstream applications used in the field of molecular biology. Whole blood samples are one of the main sources used to obtain DNA, and there are many different protocols available to perform nucleic acid extraction on such samples. These methods vary from very basic manual protocols to more sophisticated methods included in automated DNA extraction protocols. Based on the wide range of available options, it would be ideal to determine the ones that perform best in terms of cost-effectiveness and time efficiency. We have reviewed DNA extraction history and the most commonly used methods for DNA extraction from whole blood samples, highlighting their individual advantages and disadvantages. We also searched current scientific literature to find studies comparing different nucleic acid extraction methods, to determine the best available choice. Based on our research, we have determined that there is not enough scientific evidence to support one particular DNA extraction method from whole blood samples. Choosing a suitable method is still a process that requires consideration of many different factors, and more research is needed to validate choices made at facilities around the world.

'Bacon - hold the eggs' for uncontrollable nose bleeds

Almost everyone experiences a nosebleed at some time during their lives. In many cases, these nosebleeds are self-limiting, resolve without medical attention, and tend to be nothing more than a minor nuisance. However, in some instances they can be life threatening and urgent medical attention is required. One of these instances is when people who have Glanzmann's thrombasthenia (GT) experience a nosebleed...

Some comparison on whole-proteome phylogeny of large dsDNA viruses based on dynamical language approach and feature frequency profiles method

There has been a growing interest in alignment-free methods for phylogenetic analysis using complete genome data. Among them, CVTree method, feature frequency profiles method and dynamical language approach were used to investigate the whole-proteome phylogeny of large dsDNA viruses. Using the data set of large dsDNA viruses from Gao and Qi (BMC Evol. Biol. 2007), the phylogenetic results based on the CVTree method and the dynamical language approach were compared in Yu et al. (BMC Evol. Biol. 2010). In this paper, we first apply dynamical language approach to the data set of large dsDNA viruses from Wu et al. (Proc. Natl. Acad. Sci. USA 2009) and compare our phylogenetic results with those based on the feature frequency profiles method. Then we construct the whole-proteome phylogeny of the larger dataset combining the above two data sets. According to the report of The International Committee on the Taxonomy of Viruses (ICTV), the trees from our analyses are in good agreement to the latest classification of large dsDNA viruses.

Bad Eggs

Synopsis and review of the Australian feature film Bad Eggs, directed by Tony Martin.

The whole world shook : shifts in ethnic, national and heroic identities in children's fiction about 9/11

Like many other cataclysmic events September 11, a day now popularly believed to have 'changed the world', has become a topic taken up by children's writers. This thesis, titled The Whole World Shook: Ethnic, National and Heroic Identities in Children's Fiction About 9/11, examines how cultural identities are constructed within fictional texts for young people written about the attacks on the Twin Towers. It identifies three significant identity categories encoded in 9/11 books for children: ethnic identities, national identities, and heroic identities. The thesis argues that the identities formed within the selected children's texts are in flux, privileging performances of identities that are contingent on post-9/11 politics. This study is located within the field of children's literature criticism, which supports the understanding that children's books, like all texts, play a role in the production of identities. Children's literature is highly significant both in its pedagogical intent (to instruct and induct children into cultural practices and beliefs) and in its obscurity (in making the complex simple enough for children, and from sometimes intentionally shying away from difficult things). This literary criticism informed the study that the texts, if they were to be written at all, would be complex, varied and most likely as ambiguous and contradictory as the responses to the attacks on New York themselves. The theoretical framework for this thesis draws on a range of critical theories including literary theory, cultural studies, studies of performativity and postmodernism. This critical framework informs the approach by providing ways for: (i) understanding how political and ideological work is performed in children's literature; (ii) interrogating the constructed nature of cultural identities; (iii) developing a nuanced methodology for carrying out a close textual analysis. The textual analysis examines a representative sample of children's texts about 9/11, including picture books, young adult fiction, and a selection of DC Comics. Each chapter focuses on a different though related identity category. Chapter Four examines the performance of ethnic identities and race politics within a sample of picture books and young adult fiction; Chapter Five analyses the construction of collective, national identities in another set of texts; and Chapter Six does analytic work on a third set of texts, demonstrating the strategic performance of particular kinds of heroic identities. I argue that performances of cultural identities constructed in these texts draw on familiar versions of identities as well as contribute to new ones. These textual constructions can be seen as offering some certainties in increasingly uncertain times. The study finds, in its sample of books a co-mingling of xenophobia and tolerance; a binaried competition between good and evil and global harmony and national insularity; and a lauding of both the commonplace hero and the super-human. Being a recent corpus of texts about 9/11, these texts provide information on the kinds of 'selves' that appear to be privileged in the West since 2001. The thesis concludes that the shifting identities evident in texts that are being produced for children about 9/11 offer implicit and explicit accounts of what constitute good citizenship, loyalty to nation and community, and desirable attributes in a Western post-9/11 context. This thesis makes an original contribution to the field of children's literature by providing a focussed and sustained analysis of how texts for children about 9/11 contribute to formations of identity in these complex times of cultural unease and global unrest.

Porosity: Establishing the relationship between drying parameters and dried food quality [SpringerBriefs in Food, Health, and Nutrition]

The preservation technique of drying offers a significant increase in the shelf life of food materials, along with the modification of quality attributes due to simultaneous heat and mass transfer. Variations in porosity are just one of the microstructural changes that take place during the drying of most food materials. Some studies found that there may be a relationship between porosity and the properties of dried foods. However, no conclusive relationship has yet been established in the literature. This paper presents an overview of the factors that influence porosity, as well as the effects of porosity on dried food quality attributes. The effect of heat and mass transfer on porosity is also discussed along with porosity development in various drying methods. After an extensive review of the literature concerning the study of porosity, it emerges that a relationship between process parameters, food qualities, and sample properties can be established. Therefore, we propose a hypothesis of relationships between process parameters, product quality attributes, and porosity.

Effect of cell wall properties on porosity and shrinkage of dried apple

Water removal during drying depends on the pathway of water migration from food materials. Moreover, the water removal rate also depends on the characteristics of the cell wall of plant tissue. In this study, the influence of cell wall properties (such as moisture distribution, stiffness, thickness and cell dimension) on porosity and shrinkage of dried product was investigated. Cell wall stiffness depends on a complex combination of plant cell microstructure, composition of food materials and the water-holding capacity of the cell. In this work, a preliminary investigation of the cell wall properties of apple was conducted in order to predict changes of porosity and shrinkage during drying. Cell wall characteristics of two types of apple (Granny Smith and Red Delicious) were investigated under convective drying to correlate with porosity and shrinkage. A scanning electron microscope (SEM), 2kN Intron, pycnometer and ImageJ software were used in order to measure and analyse cell characteristics, water holding capacity of cell walls, porosity and shrinkage. The cell firmness of the Red Delicious apple was found to be higher than for Granny Smith apples. A remarkable relationship was observed between cell wall characteristics when compare with heat and mass transfer characteristics. It was also found that the evolution of porosity and shrinkage are noticeably influenced by the nature of the cell wall during convective drying. This study has revealed a better understanding of porosity and the shrinkage of dried food at microscopy (cell) level, and will provide better insights to attain energy-effective drying processes and improved quality of dried foods.

A micro-level investigation of the solid displacement method for porosity determination of dried food

Porosity is one of the key parameters of the macroscopic structure of porous media, generally defined as the ratio of the free spaces occupied (by the volume of air) within the material to the total volume of the material. Porosity is determined by measuring skeletal volume and the envelope volume. Solid displacement method is one of the inexpensive and easy methods to determine the envelope volume of a sample with an irregular shape. In this method, generally glass beads are used as a solid due to their uniform size, compactness and fluidity properties. The smaller size of the glass beads means that they enter into the open pores which have a larger diameter than the glass beads. Although extensive research has been carried out on porosity determination using displacement method, no study exists which adequately reports micro-level observation of the sample during measurement. This study set out with the aim of assessing the accuracy of solid displacement method of bulk density measurement of dried foods by micro-level observation. Solid displacement method of porosity determination was conducted using a cylindrical vial (cylindrical plastic container) and 57 µm glass beads in order to measure the bulk density of apple slices at different moisture contents. A scanning electron microscope (SEM), a profilometer and ImageJ software were used to investigate the penetration of glass beads into the surface pores during the determination of the porosity of dried food. A helium pycnometer was used to measure the particle density of the sample. Results show that a significant number of pores were large enough to allow the glass beads to enter into the pores, thereby causing some erroneous results. It was also found that coating the dried sample with appropriate coating material prior to measurement can resolve this problem.

Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis

Systems-level identification and analysis of cellular circuits in the brain will require the development of whole-brain imaging with single-cell resolution. To this end, we performed comprehensive chemical screening to develop a whole-brain clearing and imaging method, termed CUBIC (clear, unobstructed brain imaging cocktails and computational analysis). CUBIC is a simple and efficient method involving the immersion of brain samples in chemical mixtures containing aminoalcohols, which enables rapid whole-brain imaging with single-photon excitation microscopy. CUBIC is applicable to multicolor imaging of fluorescent proteins or immunostained samples in adult brains and is scalable from a primate brain to subcellular structures. We also developed a whole-brain cell-nuclear counterstaining protocol and a computational image analysis pipeline that, together with CUBIC reagents, enable the visualization and quantification of neural activities induced by environmental stimulation. CUBIC enables time-course expression profiling of whole adult brains with single-cell resolution.