Optimization of ClpXP activity and protein synthesis in an E. coli extract-based cell-free expression system.

Protein degradation is a fundamental process in all living cells and is essential to remove both damaged proteins and intact proteins that are no longer needed by the cell. We are interested in creating synthetic genetic circuits that function in a cell-free expression system. This will require not only an efficient protein expression platform but also a robust protein degradation system in cell extract. Therefore, we purified and tested the activity of E. coli ClpXP protease in cell-free transcription-translation (TX-TL) systems that used E. coli S30 cell extract. Surprisingly, our studies showed that purified ClpXP added to the TX-TL system has very low proteolytic activity. The low activity of ClpXP was correlated with the rapid consumption of adenosine triphosphate (ATP) in cell extract. We improved the activity of ClpXP in cell extract by adding exogenous ATP and an energy regeneration system. We then established conditions for both protein synthesis, and protein degradation by ClpXP to occur simultaneously in the TX-TL systems. The optimized conditions for ClpXP activity will be useful for creating tunable synthetic genetic circuits and in vitro synthetic biology

Elastic Pekeris waveguide normal mode solution comparisons against laboratory data

Following the derivation presented by Press and Ewing [Geophysics 15, 426-446 (1950)], a normal mode solution for the Pekeris waveguide problem with an elastic bottom is outlined. The analytic solution is benchmarked against data collected in an experiment performed at the Naval Research Laboratory [Collis et al., J. Acoust. Soc. Am. 122, 1987-1993 (2007)]. Comparisons reveal a close match between the analytic solution and experimental data. Results are strongly dependent on the accuracy of the horizontal wavenumbers for the modes, and horizontal wavenumber spectra are compared against those from the experimental data. (C) 2012 Acoustical Society of AmericaNational Science Foundation Division of Graduate Education [DGE-0638719

Virtual Rituals: Community, Emotion, and Ritual in Massive Multiplayer Online Role-playing Games—A Quantitative Test and Extension of Structural Ritualization Theory

Millions of people worldwide immerse themselves in massive multiplayer online role-playing games (MMORPGs). These games generate large, diverse communities that engage in rituals within the game, completing missions or quests. What role do these MMORPG rituals play in commitment to these gaming communities? To address this question, we extend structural ritualization theory to explain the impact of ritual events and emotion on commitment to community in the game World of Warcraft. Our findings suggest that players focused on inanimate resources are less committed than players who focus on social aspects of the ritual events inside the game. We also find that emotional investment is a good predictor of commitment to community

The Treadmill of Information: Development of the Information Society and Carbon Dioxide Emissions

The world is facing a crisis of global warming due to the release of CO2 and other greenhouse gasses by human activities. Many scholars and stakeholders argue that information and communication technology (ICT) development will mitigate CO2 emissions. Advocacy of technological solutions to CO2 mitigation is consistent with ecological modernization theory\u27s assertion that reflexive societies will modernize sustainably. In contrast, we define the “treadmill of information” as the unique contribution of ICT development to environmental degradation. We examine the impact of ICT development on total CO2 emissions and source-sector emissions from electricity, buildings, manufacturing, and transportation using a multilevel growth model for panel data from 113 countries split into the world, developed country, and less-developed-country samples. We find that the level of fixed telephone development is a strong predictor of higher CO2 emissions in less-developed countries, while internet use predicts higher CO2 emissions in developed countries. The effect of mobile telephone development is not significant. Thus, it appears that ICTs are not having an ameliorative effect on global warming as expected by ecological modernization theorists, and instead reinforce the treadmill of production\u27s negative effect

An Interdisciplinary Approach to Experiential Learning in Cyberbiosecurity and Agriculture Through Workforce Development

Cyberbiosecurity and workforce development in agriculture and the life sciences (ALS) is a growing area of need in the curriculum in higher education. Students that pursue majors related to ALS often do not include training in cyber-related concepts or expose the ‘hidden curriculum’ of seeking internships and jobs. Exposing students through workforce development training and hands-on engagement with industry professionals can provide learning opportunities to bridge the two and is an area of growth and demand as the workforce evolves. The objectives of this work were 1) to learn key concepts in cybersecurity, including data security, visualization, and analysis, to name a few, through class activities and engagement with professional partners and 2) to understand what knowledge students gained from participating in the course could transfer over for when they enter the workforce. Three themes emerged from the study where students, through direct engagement with industry partners, gained more insight about the industry applicable to their studies; they established work environment expectations for entering internships and official job placements and established ways in which the workforce development training informed their future careers

Processing carbon nanotubes with holographic optical tweezers

We report the first demonstration that carbon nanotubes can be trapped and manipulated by optical tweezers. This observation is surprising because individual nanotubes are substantially smaller than the wavelength of light, and thus should not be amenable to optical trapping. Even so, nanotube bundles, and perhaps even individual nanotubes, can be transported at high speeds, deposited onto substrates, untangled, and selectively ablated, all with visible light. The use of holographic optical tweezers, capable of creating hundreds of independent traps simultaneously, suggests opportunities for highly parallel nanotube processing with light.Comment: 3 pages, 1 figur

Intra-Oral Colonization of Macaque Monkeys by Actinobacillus Actinomycetemcomitans

Actinobacillus actinomycetemcomitans was acquired by captive Macaca fascicularis 3 to 6 months after birth, and all monkeys aged over 6 months harbored detectable levels. This microorganism was most frequently isolated from the gingival plaque of the incisor (and other) teeth compared with other oral sites. Strains were leukotoxic by bioassay and Western blot analysis. Antibodies in macaque serum contained neutralized the leukotoxin of a human A. actinomycetemcomitans strain. High titres of maternal neutralizing anti-leukotoxin antibodies were detected in neonates; the titre then fell rapidly so that by 6 months the antibody titer was zero. Antileukotoxin antibody production was detected after 6 months of age, rapidly reaching a high level within 2 years after birth. The presence of leukotoxic strains of A. actinomycetemcomitans in the gingival region did not appear to be correlated with an increase in susceptibility to periodontal disease

Western Washington University Alumni Attitudes and Perceptions of Their Undergraduate Experiences - 1987 and 1989

A summary of findings from two surveys of WWU alumni (1987 and 1989)