Resolving genetic functions within microbial populations: In situ analyses using rRNA and mRNA stable isotope probing coupled with single-cell raman-fluorescence in situ hybridization

Prokaryotes represent one-half of the living biomass on Earth, with the vast majority remaining elusive to culture and study within the laboratory. As a result, we lack a basic understanding of the functions that many species perform in the natural world. To address this issue, we developed complementary population and single-cell stable isotope (C-13)-linked analyses to determine microbial identity and function in situ. We demonstrated that the use of rRNA/mRNA stable isotope probing (SIP) recovered the key phylogenetic and functional RNAs. This was followed by single-cell physiological analyses of these populations to determine and quantify in situ functions within an aerobic naphthalene-degrading groundwater microbial community. Using these culture-independent approaches, we identified three prokaryote species capable of naphthalene biodegradation within the groundwater system: two taxa were isolated in the laboratory (Pseudomonas fluorescens and Pseudomonas putida), whereas the third eluded culture (an Acidovorax sp.). Using parallel population and single-cell stable isotope technologies, we were able to identify an unculturable Acidovorax sp. which played the key role in naphthalene biodegradation in situ, rather than the culturable naphthalene-biodegrading Pseudomonas sp. isolated from the same groundwater. The Pseudomonas isolates actively degraded naphthalene only at naphthalene concentrations higher than 30 mu M. This study demonstrated that unculturable microorganisms could play important roles in biodegradation in the ecosystem. It also showed that the combined RNA SIP-Raman-fluorescence in situ hybridization approach may be a significant tool in resolving ecology, functionality, and niche specialization within the unculturable fraction of organisms residing in the natural environment.

Optical coherence tomography is a valuable tool in the study of the effects of microneedle geometry on skin penetration characteristics and in-skin dissolution.

In this study, we used optical coherence tomography (OCT) to extensively investigate, for the first time, the effect that microneedle (MN) geometry (MN height, and MN interspacing) and force of application have upon penetration characteristics of soluble poly(methylvinylether-co-maleic anhydride, PMVE/MA) MN arrays into neonatal porcine skin in vitro. The results from OCT investigations were then used to design optimal and suboptimal MN-based drug delivery systems and evaluate their drug delivery profiles cross full thickness and dermatomed neonatal porcine skin in vitro. It was found that increasing the force used for MN application resulted in a significant increase in the depth of penetration achieved within neonatal porcine skin. For example, MN of 600 µm height penetrated to a depth of 330 µm when inserted at a force of 4.4 N/array, while the penetration increased significantly to a depth of 520 µm, when the force of application was increased to 16.4 N/array. At an application force of 11.0 N/array it was found that, in each case, increasing MN height from 350 to 600 µm to 900 µm led to a significant increase in the depth of MN penetration achieved. Moreover, alteration of MN interspacing had no effect upon depth of penetration achieved, at a constant MN height and force of application. With respect to MN dissolution, an approximate 34% reduction in MN height occurred in the first 15 min, with only 17% of the MN height remaining after a 3-hour period. Across both skin models, there was a significantly greater cumulative amount of theophylline delivered after 24 h from an MN array of 900 µm height (292.23 ± 16.77 µg), in comparison to an MN array of 350 µm height (242.62 ± 14.81 µg) (p < 0.001). Employing full thickness skin significantly reduced drug permeation in both cases. Importantly, this study has highlighted the effect that MN geometry and application force have upon the depth of penetration into skin. While it has been shown that MN height has an important role in the extent of drug delivered across neonatal porcine skin from a soluble MN array, further studies to evaluate the full significance of MN geometry on MN mediated drug delivery are now underway. The successful use of OCT in this study could prove to be a key development for polymeric MN research, accelerating their commercial exploitation.

Translating ‘gaytown’: the collision of global and local in bringing Australian queer play Bison to Belfast.

Sydney playwright Lachlan Philpott’s Bison (2000/2009) is immersed in a sweaty, summery Antipodean scene of bronzed and toned bodies. It is located in the flora and fauna of gum trees and biting ants. Yet, despite this, it could be argued that at its heart it is not a specifically Australian site, but an all-too translatable scene that seems to be played out in gay clubs, bars, chatrooms and saunas around the Western world: men repeating patterns, looking for sex or love; checking out bodies, craving perfection; avoiding, and occasionally seeking, disease. At least, that was my assumption when I decided to direct the play in Belfast, Northern Ireland, in 2009. Philpott came to Belfast to workshop the play with the actors and, as a group, we restructured the play and tried to find a way to ‘de-Australianise’ it without necessarily placing it in a new geographical place - Northern Ireland - through linguistic clues in the text. As Philpott put it: ‘Let’s not make this play about Belfast or Sydney or London or anywhere because it is not a fair reflection of these scenes. Maybe we should just identify the generic elements of this world and then make Bison a play that reflects gaytown – because the rituals are all the same in Western society’. The experience of doing the play in Belfast made clear, however, that ideas of a global gay identity/experience –though highly marketed – fail to account for the vastly different situations of embodied gay experience. And the Northern Irish gay experience, while it has imported the usual ‘generic’ tropes of gayness, sits within a specific cultural context in which the farsighted legislation on equality for gays (imposed by either London or the EU) vastly outstrips wider societal thinking. For many in Northern Ireland, erstwhile MP Iris Robinson’s comments about homosexuality being an ‘abomination’ were a reason to support her, rather than to reject her. For me, the comments were the catalyst to doing Bison in Belfast.

Welfare epidemiology as a tool to assess the welfare impact of inherited defects on the pedigree dog population

The effect that breed standards and selective breeding practices have on the welfare of pedigree dogs has recently come under scrutiny from both the general public and scientific community. Recent research has suggested that breeding for particular aesthetic traits, such as tightly curled tails, highly domed skulls and short muzzles predisposes dogs with these traits to certain inherited defects, such as spina bifida, syringomyelia and brachycephalic airway obstruction syndrome, respectively. Further to this, there is a very large number of inherited diseases that are not related to breed standards, which are thought to be prevalent, partly as a consequence of inbreeding and restricted breeding pools. Inherited diseases, whether linked to conformation or not, have varying impact on the individuals affected by them, and affect varying proportions of the pedigree dog population. Some diseases affect few breeds but are highly prevalent in predisposed breeds. Other diseases affect many breeds, but have low prevalence within each breed. In this paper, we discuss the use of risk analysis and severity diagrams as means of mapping the overall problem of inherited disorders in pedigree dogs and, more specifically, the welfare impact of specific diseases in particular breeds.

Measuring health-related quality of life after stroke: A brief tool.

Routine assessment of health-related quality of life (HRQoL) can be time consuming and burdensome for a person with stroke. Therefore the aim of this study was to develop and test a brief instrument for assessing HRQoL among people with stroke. The Quality of Life after Stroke Scale (QLASS) was constructed from items within the Quality of Life Index-Stroke Version and the Chronic Respiratory Disease Questionnaire. It was administered to 92 people with stroke at three points in time: immediately after discharge from hospital, 6 months and 12 months later. Results suggest that the QLASS has 19 items which represent three factors: emotional functioning, mastery and fatigue which correlate with valid measures of health status and activities of daily living. The QLASS is proposed as a brief, valid HRQoL tool for use among people with stroke.

Whose play is it anyway?: Theatre Studies, Translation Studies and translation for the stage

This article explores the historical neglect of translation as a consideration in the study and practice of theatre in the United States and Europe. While the study of literature is fairly strictly divided between English-language and Comparative Literature departments, theatre and drama have shown little concern about language as a barrier to reception of the dramatic text. Arguably, this discrepancy may be traced to a fundamental gap between the perceived status of the novel as a completed work of art and the playtext as work of art in progress, waiting to find its completion in performance.