947 resultados para site-directed mutagenesis
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We propose a new kind of asymmetric mutual authentication from passwords with stronger privacy against malicious servers, lest they be tempted to engage in “cross-site user impersonation” to each other. It enables a person to authenticate (with) arbitrarily many independent servers, over adversarial channels, using a memorable and reusable single short password. Beside the usual PAKE security guarantees, our framework goes to lengths to secure the password against brute-force cracking from privileged server information.
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Nick Shackleton’s research on piston cores from the Iberian margin highlighted the importance of this region for providing high-fidelity records of millennial-scale climate variability, and for correlating climate events from the marine environment to polar ice cores and European terrestrial sequences. During the Integrated Ocean Drilling Program (IODP) Expedition 339, we sought to extend the Iberian margin sediment record by drilling with the D/V JOIDES Resolution. Five holes were cored at Site U1385 using the advanced piston corer (APC) system to a maximum depth of ∼ 155.9 m below sea floor (m b.s.f.). Immediately after the expedition, cores from all holes were analyzed by core scanning X-ray fluorescence (XRF) at 1 cm spatial resolution. Ca/Ti data were used to accurately correlate from hole-to-hole and construct a composite spliced section, containing no gaps or disturbed intervals to 166.5 m composite depth (mcd). A low-resolution (20 cm sample spacing) oxygen isotope record confirms that Site U1385 contains a continuous record of hemipelagic sedimentation from the Holocene to 1.43 Ma (Marine Isotope Stage 46). The sediment profile at Site U1385 extends across the middle Pleistocene transition (MPT) with sedimentation rates averaging ∼ 10 cm kyr−1. Strongprecession cycles in colour and elemental XRF signals provide a powerful tool for developing an orbitally tuned reference timescale. Site U1385 is likely to become an important type section for marine–ice–terrestrial core correlations and the study of orbital- and millennial-scale climate variability.
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Contemporary lipidomics protocols are dependent on conventional tandem mass spectrometry for lipid identification. This approach is extremely powerful for determining lipid class and identifying the number of carbons and the degree of unsaturation of any acyl-chain substituents. Such analyses are however, blind to isomeric variants arising from different carbon carbon bonding motifs within these chains including double bond position, chain branching, and cyclic structures. This limitation arises from the fact that conventional, low energy collision-induced dissociation of even-electron lipid ions does not give rise to product ions from intrachain fragmentation of the fatty acyl moieties. To overcome this limitation, we have applied radical-directed dissociation (RDD) to the study of lipids for the first time. In this approach, bifunctional molecules that contain a photocaged radical initiator and a lipid-adducting group, such as 4-iodoaniline and 4-iodobenzoic acid, are used to form noncovalent complexes (i.e., adduct ions) with a lipid during electrospray ionization. Laser irradiation of these complexes at UV wavelengths (266 nm) cleaves the carbon iodine bond to liberate a highly reactive phenyl radical. Subsequent activation of the nascent radical ions results in RDD with significant intrachain fragmentation of acyl moieties. This approach provides diagnostic fragments that are associated with the double bond position and the positions of chain branching in glycerophospholipids, sphingomyelins and triacylglycerols and thus can be used to differentiate isomeric lipids differing only in such motifs. RDD is demonstrated for well-defined lipid standards and also reveals lipid structural diversity in olive oil and human very-low density lipoprotein.
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The appropriateness of default investment options in participant-directed retirement plans like 401(k) has been in sharp focus given that most participants fail to nominate an investment option to direct their contributions. In United States (US), prior to the Pension Protection Act (PPA) of 2006, plan fiduciaries often selected a money market fund as the default option. Whilst this ‘low risk and low return’ investment option was considered to be a ‘safe’ choice by many fiduciaries who were fearful of litigation risk, it was heavily criticized for resulting in inadequate wealth at retirement, particularly when retirees were living much longer and facing inflation risk (see, for example, Viceira, 2008; Skinner, 2009)...
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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.
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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.
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This study models young people's moderate drinking decision-making using the Model of Goal-Directed Behaviour (MGB), thus presenting insights into young people's desires and intentions to drink responsibly. Testing the applicability of the MGB to quantitatively analyse responsible drinking, the explanatory sphere of the MGB is extended. An online survey resulted in 1522 completed questionnaires from respondents aged between 18 and 25 years. Collected data were analysed with structural equation modelling (SEM) using SPSS AMOS21 (IBM, New York, NY, USA) software. The key finding of this study is that an individual's desire to drink moderately is the most important predictor of young people's responsible drinking intentions. Our use of MGB provides further evidence that there is a strong distinction between consumer desires and intentions.
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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
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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.
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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.
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We have developed a Hierarchical Look-Ahead Trajectory Model (HiLAM) that incorporates the firing pattern of medial entorhinal grid cells in a planning circuit that includes interactions with hippocampus and prefrontal cortex. We show the model’s flexibility in representing large real world environments using odometry information obtained from challenging video sequences. We acquire the visual data from a camera mounted on a small tele-operated vehicle. The camera has a panoramic field of view with its focal point approximately 5 cm above the ground level, similar to what would be expected from a rat’s point of view. Using established algorithms for calculating perceptual speed from the apparent rate of visual change over time, we generate raw dead reckoning information which loses spatial fidelity over time due to error accumulation. We rectify the loss of fidelity by exploiting the loop-closure detection ability of a biologically inspired, robot navigation model termed RatSLAM. The rectified motion information serves as a velocity input to the HiLAM to encode the environment in the form of grid cell and place cell maps. Finally, we show goal directed path planning results of HiLAM in two different environments, an indoor square maze used in rodent experiments and an outdoor arena more than two orders of magnitude larger than the indoor maze. Together these results bridge for the first time the gap between higher fidelity bio-inspired navigation models (HiLAM) and more abstracted but highly functional bio-inspired robotic mapping systems (RatSLAM), and move from simulated environments into real-world studies in rodent-sized arenas and beyond.
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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.
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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.
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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.
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High voltage powerlines may give rise to corona breakdown, resulting in the release of large concentrations of charged ions into the surrounding environment. These ions quickly attach to aerosols and the resulting charged particles are carried by prevalent winds. This paper describes a study carried out at a site near an overhead double circuit ac transmission voltage powerline to investigate factors that control the rate at which charged particles are produced, and to determine the total particle number concentrations, total particle charge concentrations and vertical dc electric fields in the proximity of the line. Measured mean values of these three parameters at a perpendicular distance of 50m from the line were 1.8 x 103 particle cm-3, 518 ions cm3 and 520 V m-1 respectively. The net electric charge was positive and the electric field was directed downwards. These parameters were correlated with each other and monitored at four different distances from the line. Effects of meteorological parameters such as wind speed and wind direction were also investigated.