Rapid identification of cytochrome P450cam variants by in vivo screening of active site libraries

The selection of cytochrome P450 enzymes from large variant libraries, and the subsequent use of these enzymes in preparative scale biotransformations, remains a formidable challenge due to the complexities of the associated electron transport systems. Here, a powerful approach for the generation and screening of P450cam libraries for new function is presented that is both flexible and robust. A targeted library was generated wherein only the P450cam active-site amino acids Y96 and F98 were fully randomized and biotransformations, using a novel P450cam whole-cell system, were screened by GC–MS for the hydroxylation of diphenylmethane. One in 50 of the reactions screened, including 16 different variants, produced 4-hydroxydiphenylmethane with up to 92% conversion observed in the case of the Y96A variant. These results demonstrate a primary example of the screening of P450cam libraries in a format that is compatible with extension to preparative scale reactions.

Revolving kaleidoscope : site-specificity and the part object in contemporary installation art

This practice-led research project explores the possibilities for restaging and reconfiguring contemporary art installations in multiple and different locations. By exploring ideas and art that demonstrate a kaleidoscopic approach to creative practice, this project examines how analysing artists' particular processes can achieve new understandings and experiences of installation art. This project achieves this through reflection on, and analysis of creative works made throughout the research, and a critical examination of contemporary art practices.

A dynamic Web agent for verifying the security and integrity of a Web site's contents

To harness safe operation of Web-based systems in Web environments, we propose an SSPA (Server-based SHA-1 Page-digest Algorithm) to verify the integrity of Web contents before the server issues an HTTP response to a user request. In addition to standard security measures, our Java implementation of the SSPA, which is called the Dynamic Security Surveillance Agent (DSSA), provides further security in terms of content integrity to Web-based systems. Its function is to prevent the display of Web contents that have been altered through the malicious acts of attackers and intruders on client machines. This is to protect the reputation of organisations from cyber-attacks and to ensure the safe operation of Web systems by dynamically monitoring the integrity of a Web site's content on demand. We discuss our findings in terms of the applicability and practicality of the proposed system. We also discuss its time metrics, specifically in relation to its computational overhead at the Web server, as well as the overall latency from the clients' point of view, using different Internet access methods. The SSPA, our DSSA implementation, some experimental results and related work are all discussed

Late Pleistocene stratigraphy of IODP Site U1396 and compiled chronology offshore of south and south west Montserrat, Lesser Antilles

Marine sediments around volcanic islands contain an archive of volcaniclastic deposits, which can be used to reconstruct the volcanic history of an area. Such records hold many advantages over often incomplete terrestrial datasets. This includes the potential for precise and continuous dating of intervening sediment packages, which allow a correlatable and temporally-constrained stratigraphic framework to be constructed across multiple marine sediment cores. Here, we discuss a marine record of eruptive and mass-wasting events spanning ~250 ka offshore of Montserrat, using new data from IODP Expedition 340, as well as previously collected cores. By using a combination of high-resolution oxygen isotope stratigraphy, AMS radiocarbon dating, biostratigraphy of foraminifera and calcareous nannofossils and clast componentry, we identify five major events at Soufriere Hills volcano since 250 ka. Lateral correlation of these events across sediment cores collected offshore of the south and south west of Montserrat, have improved our understanding of the timing, extent and associations between events in this area. Correlations reveal that powerful and potentially erosive density-currents travelled at least 33 km offshore, and demonstrate that marine deposits, produced by eruption-fed and mass-wasting events on volcanic islands, are heterogeneous in their spatial distribution. Thus, multiple drilling/coring sites are needed to reconstruct the full chronostratigraphy of volcanic islands. This multidisciplinary study will be vital to interpreting the chaotic records of submarine landslides at other sites drilled during Expedition 340 and provides a framework that can be applied to the stratigraphic analysis of sediments surrounding other volcanic islands.

Rapid on-site evaluation of fine needle aspiration specimens by cytology scientists : a review of 3032 specimens

Objectives To determine: (1) the accuracy of cytology scientists at assessing specimen adequacy by rapid on-site evaluation (ROSE) at fine needle aspiration (FNA) cytology collections; and (2) whether thyroid FNA with ROSE has lower inadequacy rates than non-attended FNAs. Methods The ROSE of adequacy for 3032 specimens from 17 anatomical sites collected over a 20-month period was compared with the final report assessment of adequacy. ROSE was performed by 19 cytology scientists. The report profile for 1545 thyroid nodules with ROSE was compared with that for 1536 consecutive non-ROSE thyroid FNAs reported by the same cytopathologists during the study period. Results ROSE was adequate in 75% (2276/3032), inadequate in 12% (366/3032) and in 13% (390/3032) no opinion was rendered. Of the 2276 cases assessed as adequate by ROSE, 2268 (99.6%) were finally reported as adequate for assessment; eight specimens had adequacy downgraded on the final report. Fifty eight per cent of cases with a ROSE assessment of inadequate were reported as adequate (212/366), whereas 93% (363/390) with no opinion rendered were reported as adequate. The overall final report adequacy rate for the 3032 specimens was 94% (2843/3032). Confirmation of a ROSE of adequacy at reporting was uniformly high amongst the 19 scientists, ranging from 98% to 100%. The inadequacy rate for thyroid FNAs with ROSE (6%) was significantly (P < 0.0001) lower than for non-ROSE thyroid FNAs (17%). A significantly (P = 0.02) higher proportion of adequate ROSE thyroid specimens was reported with abnormalities, compared with non-ROSE thyroid collections. Conclusions Cytology scientists are highly accurate at determining specimen adequacy at ROSE for a wide range of body sites. ROSE of thyroid FNAs can significantly reduce inadequate reports.

A robust methodological approach for mine site water accounting

Over the past decade, the mining industry has come to recognise the importance of water both to itself and to others. Water accounting is a formalisation of this importance that quantifies and communicates how water is used by individual sites and the industry as a whole. While there are a number of different accounting frameworks that could be used within the industry, the Minerals Council of Australia’s (MCA) Water Accounting Framework (WAF) is an industry-led approach that provides a consistent representation of mine site water interactions regardless of their operational, social or environmental context that allows for valid comparisons between sites and companies. The WAF contains definitions of offsite water sources and destinations and onsite water use, a methodology for applying the definitions and a set of metrics to measure site performance. The WAF is comprised of two models: the Input-Output Model, which represents the interactions between sites and their surrounding community and the Operational Model, which represents onsite water interactions. Members of the MCA have recently adopted the WAF’s Input-Output Model to report on their external water interactions in their Australian operations with some adopting it on a global basis. To support this adoption, there is a need for companies to better understand how to implement the WAF in their own operations. Developing a water account is non-trivial, particularly for sites unfamiliar with the WAF or for sites with the need to represent unusual features. This work describes how to build a water account for a given site using the Input-Output Model with an emphasis on how to represent challenging situations.

Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination

RNA polymerase II (pol II) transcription termination requires co‐transcriptional recognition of a functional polyadenylation signal, but the molecular mechanisms that transduce this signal to pol II remain unclear. We show that Yhh1p/Cft1p, the yeast homologue of the mammalian AAUAAA interacting protein CPSF 160, is an RNA‐binding protein and provide evidence that it participates in poly(A) site recognition. Interestingly, RNA binding is mediated by a central domain composed of predicted β‐propeller‐forming repeats, which occurs in proteins of diverse cellular functions. We also found that Yhh1p/Cft1p bound specifically to the phosphorylated C‐terminal domain (CTD) of pol II in vitro and in a two‐hybrid test in vivo. Furthermore, transcriptional run‐on analysis demonstrated that yhh1 mutants were defective in transcription termination, suggesting that Yhh1p/Cft1p functions in the coupling of transcription and 3′‐end formation. We propose that direct interactions of Yhh1p/Cft1p with both the RNA transcript and the CTD are required to communicate poly(A) site recognition to elongating pol II to initiate transcription termination.

Improving the accuracy of mine site water balances through improved estimation of runoff volumes

A mine site water balance is important for communicating information to interested stakeholders, for reporting on water performance, and for anticipating and mitigating water-related risks through water use/demand forecasting. Gaining accuracy over the water balance is therefore crucial for sites to achieve best practice water management and to maintain their social license to operate. For sites that are located in high rainfall environments the water received to storage dams through runoff can represent a large proportion of the overall inputs to site; inaccuracies in these flows can therefore lead to inaccuracies in the overall site water balance. Hydrological models that estimate runoff flows are often incorporated into simulation models used for water use/demand forecasting. The Australian Water Balance Model (AWBM) is one example that has been widely applied in the Australian context. However, the calibration of AWBM in a mining context can be challenging. Through a detailed case study, we outline an approach that was used to calibrate and validate AWBM at a mine site. Commencing with a dataset of monitored dam levels, a mass balance approach was used to generate an observed runoff sequence. By incorporating a portion of this observed dataset into the calibration routine, we achieved a closer fit between the observed vs. simulated dataset compared with the base case. We conclude by highlighting opportunities for future research to improve the calibration fit through improving the quality of the input dataset. This will ultimately lead to better models for runoff prediction and thereby improve the accuracy of mine site water balances.

A comparative study of airborne particulate pollution at two bus stations of different site geography

Airborne particulate pollutant is considered to be one of the major harmful emissions produced by vehicle engines as it has been directly linked to serious health problems. Passengers spend long times at bus stations and may be exposed to high concentrations of pollution. Particle pollution at two bus stations in Brisbane, Australia were monitored. The two bus stations consisted of markedly different site geography and surroundings with one situated in a street canyon and the other elevated above ground level. The same flow of traffic operated through both stations. Real time measurements of ultrafine particle concentration, size distribution and meteorological conditions were carried out on the platform continuously over several days. The results showed that the particle number concentrations were significantly different at the two stations, suggesting that the layout of site geometry and surroundings was a dominant determining factor through the injection of fresh air into the station platforms and the rates of dilution.

Antisocial behaviour orders : Unanticipated directions in social network site development

Abstract: Social network technologies, as we know them today have become a popular feature of everyday life for many people. As their name suggests, their underlying premise is to enable people to connect with each other for a variety of purposes. These purposes however, are generally thought of in a positive fashion. Based on a multi-method study of two online environments, Habbo Hotel and Second Life, which incorporate social networking functionality, we she light on forms of what can be conceptualized as antisocial behaviours and the rationales for these. Such behaviours included: scamming, racist/homophobic attacks, sim attacks, avatar attacks, non-conformance to contextual norms, counterfeiting and unneighbourly behaviour. The rationales for sub behaviours included: profit, fun, status building, network disruption, accidental acts and prejudice. Through our analysis we are able to comment upon the difficulties of defining antisocial behaviour in such environments, particularly when such environments are subject to interpretation vis their use and expected norms. We also point to the problems we face in conducting our public and private lives given the role ICTs are playing in the convergence of these two spaces and also the convergence of ICTs themselves.

‘He has a way gayer Facebook than I do’ : investigating sexual identity disclosure and context collapse on a social networking site

This study extends previous research into social networking sites (SNSs) as environments that often reduce spatial, temporal, and social boundaries, which can result in collapsed contexts for social situations. Context collapse was investigated through interviews and Facebook walkthroughs with 27 LGBTQ young people in the United Kingdom. Since diverse sexualities are often stigmatized, participants’ sexual identity disclosure decisions were shaped by both the social conditions of their online networks and the technological architecture of SNSs. Context collapse was experienced as an event through which individuals intentionally redefined their sexual identity across audiences or managed unintentional disclosure. To prevent unintentional context collapse, participants frequently reinstated contexts through tailored performances and audience separation. These findings provide insight into stigmatized identity performances in networked publics while situating context collapse within a broader understanding of impression management, which paves the way for future research exploring the identity implications of everyday SNS use.