Lentiviral vector design using alternative RNA export elements

<p>Abstract</p> <p>Background</p> <p>Lentiviral vectors have been designed with complex RNA export sequences in both the integrating and packaging plasmids in order to co-ordinate efficient vector production. Recent studies have attempted to replace the existing complex rev/RRE system with a more simplistic RNA export system from simple retroviruses to make these vectors in a rev-independent manner.</p> <p>Results</p> <p>Towards this end, lentiviral transfer plasmids were modified with various cis-acting DNA elements that co-ordinate RNA export during viral production to determine their ability to affect the efficiency of vector titer and transduction in different immortalized cell lines in vitro. It was found that multiple copies of the constitutive transport element (CTE) originating from different simian retroviruses, including simian retrovirus type 1 (SRV-1) and type-2 (SRV-2) and Mason-Pfizer (MPV) could be used to eliminate the requirement for the rev responsive element (RRE) in the transfer and packaging plasmids with titers >10⁶T.U./mL (n = 4–8 preparations). The addition of multiple copies of the murine intracisternal type A particle, the woodchuck post-regulatory element (WPRE), or single and dual copies of the simian CTE had minimal effect on viral titer. Immortalized cell lines from different species were found to be readily transduced by VSV-G pseudotyped lentiviral vectors containing the multiple copies of the CTE similar to the findings in HeLa cells, although the simian-derived CTE were found to have a lower infectivity into murine cell lines compared to the other species.</p> <p>Conclusion</p> <p>These studies demonstrated that the rev-responsive element (RRE) could be replaced with other constitutive transport elements to produce equivalent titers using lentivectors containing the RRE sequence <it>in vitro</it>, but that concatemerization of the CTE or the close proximity of RNA export sequences was needed to enhance vector production.</p

Agent-based performance assessment tool for general aviation operations under free flight

The objective of this research is to design and demonstrate an agent-based modeling and analysis tool for evaluating General Aviation (GA) pilot situation awareness under free flight air traffic management (ATM). A computational tool is developed to assess free flight's potential effect on GA operators, by combining an agent-based representation of the overall pilot/vehicle/ATM system with quantitative modelbased metrics of pilot SA. The model's performance is demonstrated in a set of simulation trials designed to measure the pilot agent's ability to recognize and correctly assess protected zone conflicts in free flight ATM, using information available from a hypothetical cockpit display of traffic information. A set of simulations is presented to examine the effect of sensor accuracy and attention allocation on pilot awareness of protected zone conflict hazards posed by intruder aircraft. The results show that reducing sensor accuracy leads to an increase in overall SA error, and that the pilot agent divides its attention over multiple traffic hazards in proportion to each intruder's hazard potential. This attention-sharing varies dynamically as the conflict situation changes, in a manner that is consistent with intuitive expectations

An empirical cognitive model of the development of shared understanding of requirements

It is well documented that customers and software development teams need to share and refine understanding of the requirements throughout the software development lifecycle. The development of this shared understand- ing is complex and error-prone however. Techniques and tools to support the development of a shared understanding of requirements (SUR) should be based on a clear conceptualization of the phenomenon, with a basis on relevant theory and analysis of observed practice. This study contributes to this with a detailed conceptualization of SUR development as sequence of group-level state transi- tions based on specializing the Team Mental Model construct. Furthermore it proposes a novel group-level cognitive model as the main result of an analysis of data collected from the observation of an Agile software development team over a period of several months. The initial high-level application of the model shows it has promise for providing new insights into supporting SUR development

Mental models of high reliability systems

Reliable performance in complex systems is determined in part by the ade quacy with which mental models of the system capture accurately the dimen sions of system coupling and system complexity. Failure to register coupling and complexity leads the observer to intervene into an imagined technology that does not exist and to convert opportunities for error into actual errors. To decrease the frequency with which this conversion occurs, people can make their models more complex or the systems they monitor less complex. Neither type of change is as daunting as it may appear, and this is illustrated by an analysis of the mental model and system design associated with the invasion of Grenada.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/68652/2/10.1177_108602668900300203.pd

The Role of Human-Automation Consensus in Multiple Unmanned Vehicle Scheduling

Objective: This study examined the impact of increasing automation replanning rates on operator performance and workload when supervising a decentralized network of heterogeneous unmanned vehicles. Background: Futuristic unmanned vehicles systems will invert the operator-to-vehicle ratio so that one operator can control multiple dissimilar vehicles connected through a decentralized network. Significant human-automation collaboration will be needed because of automation brittleness, but such collaboration could cause high workload. Method: Three increasing levels of replanning were tested on an existing multiple unmanned vehicle simulation environment that leverages decentralized algorithms for vehicle routing and task allocation in conjunction with human supervision. Results: Rapid replanning can cause high operator workload, ultimately resulting in poorer overall system performance. Poor performance was associated with a lack of operator consensus for when to accept the automation’s suggested prompts for new plan consideration as well as negative attitudes toward unmanned aerial vehicles in general. Participants with video game experience tended to collaborate more with the automation, which resulted in better performance. Conclusion: In decentralized unmanned vehicle networks, operators who ignore the automation’s requests for new plan consideration and impose rapid replans both increase their own workload and reduce the ability of the vehicle network to operate at its maximum capacity. Application: These findings have implications for personnel selection and training for futuristic systems involving human collaboration with decentralized algorithms embedded in networks of autonomous systems.Aurora Flight Sciences Corp.United States. Office of Naval Researc