947 resultados para site-directed mutagenesis
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The ability to automate forced landings in an emergency such as engine failure is an essential ability to improve the safety of Unmanned Aerial Vehicles operating in General Aviation airspace. By using active vision to detect safe landing zones below the aircraft, the reliability and safety of such systems is vastly improved by gathering up-to-the-minute information about the ground environment. This paper presents the Site Detection System, a methodology utilising a downward facing camera to analyse the ground environment in both 2D and 3D, detect safe landing sites and characterise them according to size, shape, slope and nearby obstacles. A methodology is presented showing the fusion of landing site detection from 2D imagery with a coarse Digital Elevation Map and dense 3D reconstructions using INS-aided Structure-from-Motion to improve accuracy. Results are presented from an experimental flight showing the precision/recall of landing sites in comparison to a hand-classified ground truth, and improved performance with the integration of 3D analysis from visual Structure-from-Motion.
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RNA polymerase III (Pol III) as well as Pol II (35S) promoters are able to drive hairpin RNA (hpRNA) expression and induce target gene silencing in plants. siRNAs of 21 nt are the predominant species in a 35S Pol II line, whereas 24- and/or 22-nucleotide (nt) siRNAs are produced by a Pol III line. The 35S line accumulated the loop of the hpRNA, in contrast to full-length hpRNA in the Pol III line. These suggest that Pol II and Pol III-transcribed hpRNAs are processed by different pathways. One Pol III transgene produced only 24-nt siRNAs but silenced the target gene efficiently, indicating that the 24-nt siRNAs can direct mRNA degradation; specific cleavage was confirmed by 59 rapid amplification of cDNA ends (RACE). Both Pol II- and Pol III-directed hpRNA transgenes induced cytosine methylation in the target DNA. The extent of methylation is not correlated with the level of 21-nt siRNAs, suggesting that they are not effective inducers of DNA methylation. The promoter of a U6 transgene was significantly methylated, whereas the promoter of the endogenous U6 gene was almost free of cytosine methylation, suggesting that endogenous sequences are more resistant to de novo DNA methylation than are transgene constructs. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 RNA Society.
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Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We created transgenic strains of Drosophila melanogaster in which the rrf-1 or ego-1 RdRp genes from C. elegans were placed under the control of the yeast GAL4 upstream activation sequence. Activation of the gene was performed by crossing these lines to flies carrying the GAL4 transgene under the control of various Drosophila enhancers. RT-PCR confirmed the successful expression of each RdRp gene. The resulting phenotypes indicated that introduction of the RdRp genes had no effect on D. melanogaster morphological development. © 2010 Springer Science+Business Media B.V.
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Abstract BACKGROUND: An examination of melanoma incidence according to anatomical region may be one method of monitoring the impact of public health initiatives. OBJECTIVES: To examine melanoma incidence trends by body site, sex and age at diagnosis or body site and morphology in a population at high risk. MATERIALS AND METHODS: Population-based data on invasive melanoma cases (n = 51473) diagnosed between 1982 and 2008 were extracted from the Queensland Cancer Registry. Age-standardized incidence rates were calculated using the direct method (2000 world standard population) and joinpoint regression models were used to fit trend lines. RESULTS: Significantly decreasing trends for melanomas on the trunk and upper limbs/shoulders were observed during recent years for both sexes under the age of 40 years and among males aged 40-59years. However, in the 60 and over age group, the incidence of melanoma is continuing to increase at all sites (apart from the trunk) for males and on the scalp/neck and upper limbs/shoulders for females. Rates of nodular melanoma are currently decreasing on the trunk and lower limbs. In contrast, superficial spreading melanoma is significantly increasing on the scalp/neck and lower limbs, along with substantial increases in lentigo maligna melanoma since the late 1990s at all sites apart from the lower limbs. CONCLUSIONS: In this large study we have observed significant decreases in rates of invasive melanoma in the younger age groups on less frequently exposed body sites. These results may provide some indirect evidence of the impact of long-running primary prevention campaigns.
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Australia’s mining boom Global demand for minerals and energy products has fuelled Australia’s recent resources boom and has led to the rapid expansion of mining projects not only in remote locations but increasingly in settled traditionally agricultural rural areas. A fundamental shift has also occurred in the provisioning of skilled and semi-skilled workers. The huge acceleration in industry demand for labour has been accompanied by the entrenchment of workforce arrangements largely dependent on fly-in, fly-out (FIFO) and drive–in, drive–out (DIDO) non-resident workers (NRWs). While NRWs are working away from their homes, they are usually accommodated in work camps or ‘villages’ for the duration of their work cycle which are normally comprised of many consecutive days of 12-hour day- and night-shifts. The health effects of this form of employment and the accompanying lifestyle is increasingly becoming contentious. Impacts on personal wellness, wellbeing and quality of life essentially remain under-researched and thus misunderstood. Sodexo in Australia Sodexo began operations in Australia in 1982, and has since become a leader in providing Quality of Life (QOL) services to businesses across the country. The 6,000 Australian employees are part of a global Sodexo team of 413,000 people. Sodexo in Australia designs, delivers and manages on-site their QOL services at 320 diverse site locations, including remote sites. Sodexo operates in a range of sectors, including the mining industry. Service plans are tailored to suit the individual needs of organisations. Sodexo Remote Sites has previously conducted unpublished research among mining workers in Australia. The results highlighted needs and expectations of Australian mining workers. Main insights about workers’ requirements were directed towards: • contacts with closest; • warm rest time around proper and varied meals; • additional services to help them better enjoy their life onsite and/or make the most of it; • organise their transportation; • promote community living; and • finding balance between professional and personal life. The brief for this current research is aimed at building upon this knowledge. Research brief Expectations for quality of life and wellness and wellbeing services are increasing dramatically. It's getting costlier and more difficult to retain valuable employees. This is particularly the case in the Australian mining sector. Given the level of interest in ensuring healthy workplaces in Australia, Sodexo has commissioned QUT to conduct a literature review. The objectives as specified by Sodexo are: Objective 1: To define the concepts of wellness and wellbeing and quality of life in Australia Objective 2: To examine how wellness and wellbeing are developed within organisations in Australia and how they impact on employee and organizational performance. More specifically, to review the literature that could be sourced about: • challenges of the mining environment; • the mining lifestyle – implications for health, wellness and daily life; • personal health and wellness of Australian mining workers; • factors affecting health in mines and perceived support for health and wellness; and • the impact of employer investment in health on perceptions and behaviour of employees. Objective 3: To determine what impact employee wellness and well-being has on the performance of mining workers. More specifically, to review the literature that could be sourced about: • impact of obesity, alcohol, tobacco use on companies; and • links between employee engagement and satisfaction and company productivity. Accordingly this review has attempted to ascertain what factors an organisation should focus on in order to reduce absenteeism and turnover and increase commitment, satisfaction, safety and productivity, with specific reference to the mining industry in Australia. The structure of the report aligns with the stated objectives in that each of the first three parts address an objective. Part IV summarises prominent issues that have arisen and offers some concluding observations and comments.
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BACKGROUND: The objective of this study was to determine whether it is possible to predict driving safety in individuals with homonymous hemianopia or quadrantanopia based upon a clinical review of neuro-images that are routinely available in clinical practice. METHODS: Two experienced neuro-ophthalmologists viewed a summary report of the CT/MRI scans of 16 participants with homonymous hemianopic or quadrantanopic field defects which provided information regarding the site and extent of the lesion and made predictions regarding whether they would be safe/unsafe to drive. Driving safety was defined using two independent measures: (1) The potential for safe driving was defined based upon whether the participant was rated as having the potential for safe driving, determined through a standardized on-road driving assessment by a certified driving rehabilitation specialist conducted just prior and (2) state recorded motor vehicle crashes (all crashes and at-fault). Driving safety was independently defined at the time of the study by state recorded motor vehicle crashes (all crashes and at-fault) recorded over the previous 5 years, as well as whether the participant was rated as having the potential for safe driving, determined through a standardized on-road driving assessment by a certified driving rehabilitation specialist. RESULTS: The ability to predict driving safety was highly variable regardless of the driving outcome measure, ranging from 31% to 63% (kappa levels ranged from -0.29 to 0.04). The level of agreement between the neuro-ophthalmologists was also only fair (kappa =0.28). CONCLUSIONS: The findings suggest that clinical evaluation of summary reports currently available neuro-images by neuro-ophthalmologists is not predictive of driving safety. Future research should be directed at identifying and/or developing alternative tests or strategies to better enable clinicians to make these predictions.
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This retrospective review examines healing in different sites on a porcine burn model; 24 pairs of burns on 18 pigs from other animal trials were selected for analysis. Each pair of burns was located on the either the cranial or the caudal part of the thoracic ribs region, on the same side of the animal. The burns were 40-50 cm(2) in size and of uniform deep-dermal partial thickness. Caudal burns healed significantly better than cranial burns, demonstrated by earlier closure of wounds, less scar formation and better cosmesis. To our knowledge, this is the first detailed study reporting that burn healing is affected by location on a porcine burn model. We recommend that similar symmetrical burns should be used for future comparative assessments of burn healing.
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Research Statement: In 2011 The State Library of Queensland in collaboration with Queensland University of Technology School of Design held a screening of six student urban films shot on location in several inner-city sites under my supervision. The films are now a permanent "exhibit" on The Edge State Library electronic site. The students were directed to explore the realist film ethos, which forms a platform for the research project, in its focus on the nonrepresentational aesthetics of the street, the unfinished and the sensory. The research demonstrates that film is a powerful instrument for the urban imaginary, for screening the city.
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Objective Recently, a number of studies have identified self-employed Protective Behavioral Strategies (PBS) as effective in decreasing the level of alcohol-related harm among young people. However, much of the published research has ignored important gender differences, such as women's increased tendency to rely on PBS that are social in nature. To further the understanding of women's PBS, the current study sought to investigate the nature and correlates of the strategies young women employ to keep their friends safe when drinking (i.e., peer-directed PBS). Method A scale measuring peer-directed PBS was developed and administered in conjunction with existing measures of alcohol consumption, personal PBS, and peer attachment. Participants consisted of 422 women aged 18–30 years, recruited among psychology students and the general public. Results Exploratory factor analysis revealed two clusters of peer-directed PBS; those that were aimed at reducing intoxication among one's friends and those that were designed to minimize alcohol-related harms. Further analysis found a positive relationship between women's tendency to implement personal and peer-directed PBS and that risky drinkers were less likely to engage in personal or peer-directed PBS (either type). Conclusion Findings indicate that personal and peer-directed PBS are related behaviors that are less frequently adopted by risky drinkers.
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A decision-making framework for image-guided radiotherapy (IGRT) is being developed using a Bayesian Network (BN) to graphically describe, and probabilistically quantify, the many interacting factors that are involved in this complex clinical process. Outputs of the BN will provide decision-support for radiation therapists to assist them to make correct inferences relating to the likelihood of treatment delivery accuracy for a given image-guided set-up correction. The framework is being developed as a dynamic object-oriented BN, allowing for complex modelling with specific sub-regions, as well as representation of the sequential decision-making and belief updating associated with IGRT. A prototype graphic structure for the BN was developed by analysing IGRT practices at a local radiotherapy department and incorporating results obtained from a literature review. Clinical stakeholders reviewed the BN to validate its structure. The BN consists of a sub-network for evaluating the accuracy of IGRT practices and technology. The directed acyclic graph (DAG) contains nodes and directional arcs representing the causal relationship between the many interacting factors such as tumour site and its associated critical organs, technology and technique, and inter-user variability. The BN was extended to support on-line and off-line decision-making with respect to treatment plan compliance. Following conceptualisation of the framework, the BN will be quantified. It is anticipated that the finalised decision-making framework will provide a foundation to develop better decision-support strategies and automated correction algorithms for IGRT.
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Background and Purpose The β1-adrenoceptor has at least two binding sites, high and low affinity sites (β1H and β1L, respectively), which mediate cardiostimulation. While β1H-adrenoceptor can be blocked by all clinically used β-blockers, β1L-adrenoceptor is relatively resistant to blockade. Thus, chronic β1L-adrenoceptor activation may mediate persistent cardiostimulation, despite the concurrent blockade of β1H-adrenoceptors. Hence, it is important to determine the potential significance of β1L-adrenoceptors in vivo, particularly in pathological situations. Experimental Approach C57Bl/6 male mice were used. Chronic (4 or 8 weeks) β1L-adrenoceptor activation was achieved by treatment, via osmotic mini pumps, with (-)-CGP12177 (10 mg·kg−1·day−1). Cardiac function was assessed by echocardiography and micromanometry. Key Results (-)-CGP12177 treatment of healthy mice increased heart rate and left ventricular (LV) contractility. (-)-CGP12177 treatment of mice subjected to transverse aorta constriction (TAC), during weeks 4–8 or 4–12 after TAC, led to a positive inotropic effect and exacerbated fibrogenic signalling while cardiac hypertrophy tended to be more severe. (-)-CGP12177 treatment of mice with TAC also exacerbated the myocardial expression of hypertrophic, fibrogenic and inflammatory genes compared to untreated TAC mice. Washout of (-)-CGP12177 revealed a more pronounced cardiac dysfunction after 12 weeks of TAC. Conclusions and Implications β1L-adrenoceptor activation provides functional support to the heart, in both normal and pathological (pressure overload) situations. Sustained β1L-adrenoceptor activation in the diseased heart exacerbates LV remodelling and therefore may promote disease progression from compensatory hypertrophy to heart failure.
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Background Prevention strategies are critical to reduce infection rates in total joint arthroplasty (TJA), but evidence-based consensus guidelines on prevention of surgical site infection (SSI) remain heterogeneous and do not necessarily represent this particular patient population. Questions/Purposes What infection prevention measures are recommended by consensus evidence-based guidelines for prevention of periprosthetic joint infection? How do these recommendations compare to expert consensus on infection prevention strategies from orthopedic surgeons from the largest international tertiary referral centers for TJA? Patients and Methods A review of consensus guidelines was undertaken as described by Merollini et al. Four clinical guidelines met inclusion criteria: Centers for Disease Control and Prevention's, British Orthopedic Association, National Institute of Clinical Excellence's, and National Health and Medical Research Council's (NHMRC). Twenty-eight recommendations from these guidelines were used to create an evidence-based survey of infection prevention strategies that was administered to 28 orthopedic surgeons from members of the International Society of Orthopedic Centers. The results between existing consensus guidelines and expert opinion were then compared. Results Recommended strategies in the guidelines such as prophylactic antibiotics, preoperative skin preparation of patients and staff, and sterile surgical attire were considered critically or significantly important by the surveyed surgeons. Additional strategies such as ultraclean air/laminar flow, antibiotic cement, wound irrigation, and preoperative blood glucose control were also considered highly important by surveyed surgeons, but were not recommended or not uniformly addressed in existing guidelines on SSI prevention. Conclusion Current evidence-based guidelines are incomplete and evidence should be updated specifically to address patient needs undergoing TJA.
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Radical-directed dissociation of gas phase ions is emerging as a powerful and complementary alternative to traditional tandem mass spectrometric techniques for biomolecular structural analysis. Previous studies have identified that coupling of 2-[(2,2,6,6-tetramethylpiperidin-1-oxyl)methyl] benzoic acid (TEMPO-Bz) to the N-terminus of a peptide introduces a labile oxygen-carbon bond that can be selectively activated upon collisional activation to produce a radical ion. Here we demonstrate that structurally-defined peptide radical ions can also be generated upon UV laser photodissociation of the same TEMPO-Bz derivatives in a linear ion-trap mass spectrometer. When subjected to further mass spectrometric analyses, the radical ions formed by a single laser pulse undergo identical dissociations as those formed by collisional activation of the same precursor ion, and can thus be used to derive molecular structure. Mapping the initial radical formation process as a function of photon energy by photodissociation action spectroscopy reveals that photoproduct formation is selective but occurs only in modest yield across the wavelength range (300-220 nm), with the photoproduct yield maximised between 235 and 225 nm. Based on the analysis of a set of model compounds, structural modifications to the TEMPO-Bz derivative are suggested to optimise radical photoproduct yield. Future development of such probes offers the advantage of increased sensitivity and selectivity for radical-directed dissociation. © 2014 the Owner Societies.
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Fatty acids are long-chain carboxylic acids that readily produce \[M - H](-) ions upon negative ion electrospray ionization (ESI) and cationic complexes with alkali, alkaline earth, and transition metals in positive ion ESI. In contrast, only one anionic monomeric fatty acid-metal ion complex has been reported in the literature, namely \[M - 2H + (FeCl)-Cl-II](-). In this manuscript, we present two methods to form anionic unsaturated fatty acid-sodium ion complexes (i.e., \[M - 2H + Na](-)). We find that these ions may be generated efficiently by two distinct methods: (1) negative ion ESI of a methanolic solution containing the fatty acid and sodium fluoride forming an \[M - H + NaF](-) ion. Subsequent collision-induced dissociation (CID) results in the desired \[M - 2H + Na](-) ion via the neutral loss of HF. (2) Direct formation of the \[M - 2H + Na](-) ion by negative ion ESI of a methanolic solution containing the fatty acid and sodium hydroxide or bicarbonate. In addition to deprotonation of the carboxylic acid moiety, formation of \[M - 2H + Na](-) ions requires the removal of a proton from the fatty acid acyl chain. We propose that this deprotonation occurs at the bis-allylic position(s) of polyunsaturated fatty acids resulting in the formation of a resonance-stabilized carbanion. This proposal is supported by ab initio calculations, which reveal that removal of a proton from the bis-allylic position, followed by neutral loss of HX (where X = F- and -OH), is the lowest energy dissociation pathway.