Destabilization of α-helical structure in solution improves bactericidal activity of antimicrobial peptides: Opposing effects on bacterial and viral targets

We have previously examined the mechanism of antimicrobial peptides on the outer membrane of vaccinia virus. Here we show that the formulation of peptides LL37 and magainin-2B amide in polysorbate 20 (Tween-20™) results in greater reductions in virus titre than formulation without detergent, and the effect is replicated by substitution of polysorbate 20 with high ionic strength buffer. In contrast, formulation with polysorbate 20 or high ionic strength buffer has the opposite effect on bactericidal activity of both peptides, resulting in lesser reductions in titre for both gram-positive and gram-negative bacteria. Circular dichroism spectroscopy shows that the differential action of polysorbate 20 and salt on the virucidal and bactericidal activities correlates with the α-helical content of peptide secondary structure in solution, suggesting that the virucidal and bactericidal activities are mediated through distinct mechanisms. The correlation of a defined structural feature with differential activity against a host-derived viral membrane and the membranes of both gram-positive and gram-negative bacteria suggests that overall helical content in solution under physiological conditions is an important feature for consideration in the design and development of candidate peptide-based antimicrobial compounds

Airborne Radar for sUAS Sense and Avoid

A primary challenge for the safe integration of small UAS operations into the National Airspace System (NAS) is traffic deconfliction, both from manned and unmanned aircraft. The UAS Traffic Management (UTM) project being conducted at the National Aeronautics and Space Administration (NASA) considers a layered approach to separation provision, ranging from segregation of operations through airspace volumes (geofences) to autonomous sense and avoid (SAA) technologies for higher risk, densely occupied airspace. Cooperative SAA systems, such as Automatic Dependent Surveillance-Broadcast (ADS-B) and/or vehicle-to-vehicle communication systems provide significant additional risk mitigation but they fail to adequately mitigate collision risks for non-cooperative (non-transponder equipped) airborne aircraft. The RAAVIN (Radar on Autonomous Aircraft to Verify ICAROUS Navigation) flight test being conducted by NASA and the Mid-Atlantic Aviation Partnership (MAAP) was designed to investigate the applicability and performance of a prototype, commercially available sUAS radar to detect and track non-cooperative airborne traffic, both manned and unmanned. The radar selected for this research was a Frequency Modulated Continuous Wave (FMCW) radar with 120 degree azimuth and 80 degree elevation field of view operating at 24.55GHz center frequency with a 200 MHz bandwidth. The radar transmits 2 watts of power thru a Metamaterial Electronically Scanning Array antenna in horizontal polarization. When the radar is transmitting, personnel must be at least 1 meter away from the active array to limit nonionizing radiation exposure. The radar physical dimensions are 18.7cm by 12.1cm by 4.1cm and it weighs less than 820 grams making it well suited for installation on small UASs. The onboard, SAA capability, known as ICAROUS, (Independent Configurable Architecture for Reliable Operations of Unmanned Systems), developed by NASA to support sUAS operations, will provide autonomous guidance using the traffic radar tracks from the onboard radar. The RAAVIN set of studies will be conducted in three phases. The first phase included outdoor, ground-based radar evaluations performed at the Virginia Techs Kentland Farm testing range in Blacksburg, VA. The test was designed to measure how well the radar could detect and track a small UAS flying in the radars field of view. The radar was used to monitor 5 test flights consisting of outbound, inbound and crossing routes at different ranges and altitudes. The UAS flown during the ground test was the Inspire 2, a quad copter weighing less than 4250 grams (10 pounds) at maximum payload. The radar was set up to scan and track targets over its full azimuthal field of view from 0 to 40 degrees in elevation. The radar was configured to eliminate tracks generated from any targets located beyond 2000 meters from the radar and moving at velocities under 1.45 meters per second. For subsequent phases of the study the radar will be integrated with a sUAS platform to evaluate its performance in flight for SAA applications ranging from sUAS to manned GA aircraft detections and tracking. Preliminary data analysis from the first outdoor ground tests showed the radar performed well at tracking the vehicle as it flew outbound and repeatedly maintained a track out to 1000 meters (maximum 1387 meters) until the vehicle slowed to a stop to reverse direction to fly inbound. As the Inspire flew inbound tracks from beyond 800 meters, a reacquisition time delay was consistently observed between when the Inspire exceeds a speed of 1.45 meters per second and when the radar indicated an inbound target was present and maintained its track. The time delay varied between 6 seconds to over 37 seconds for the inbound flights examined, and typically resulted in about a 200 meter closure distance before the Inspire track was maintained. The radar performed well at both acquiring and tracking the vehicle as it flew crossing routes out past 400 meters across the azimuthal field of view. The radar and ICAROUS software will be integrated and flown on a BFD-1400-SE8-E UAS during the next phase of the RAAVIN project. The main goal at the conclusion of this effort is to determine if this radar technology can reliably support minimum requirements for SAA applications of sUAS. In particular, the study will measure the range of vehicle detections, lateral and vertical angular errors, false and missed/late detections, and estimated distance at closest point of approach after an avoidance maneuver is executed. This last metric is directly impacted by sensor performance and indicates its suitability for the task

The Internal Circadian Clock Increases Hunger and Appetite in the Evening Independent of Food Intake and Other Behaviors

Objective: Despite the extended overnight fast, paradoxically, people are typically not ravenous in the morning and breakfast is typically the smallest meal of the day. Here we assessed whether this paradox could be explained by an endogenous circadian influence on appetite with a morning trough, while controlling for sleep/wake and fasting/feeding effects. Design and Methods We studied 12 healthy non-obese adults (6 male; age, 20–42 year) throughout a 13-day laboratory protocol that balanced all behaviors, including eucaloric meals and sleep periods, evenly across the endogenous circadian cycle. Participants rated their appetite and food preferences by visual analog scales. Results: There was a large endogenous circadian rhythm in hunger, with the trough in the biological morning (8 AM) and peak in the biological evening (8 PM; peak-to-trough amplitude=17%; P=0.004). Similarly phased significant endogenous circadian rhythms were present in appetite for sweets, salty and starchy foods, fruits, meats/poultry, food overall, and for estimates of how much food participants could eat (amplitudes 14–25%; all P < 0.05). Conclusions: In people who sleep at night, the intrinsic circadian evening peak in appetite may promote larger meals before the fasting period necessitated by sleep. Furthermore, the circadian decline in hunger across the night would theoretically counteract the fasting-induced hunger increase that could otherwise disrupt sleep

Regulation of Net1A subcellular localization by the small GTPase Rac1

Activation of Rho family small G proteins is thought to be a critical event in breast cancer development and metastatic progression. Rho protein activation is stimulated by a family of enzymes known as guanine nucleotide exchange factors (Rho GEFs). The neuroepithelioma transforming gene 1 (Net1) is a Rho GEF specific for the RhoA subfamily that is overexpressed in primary breast tumors and breast cancer cell lines. Net1 isoform expression is also required for migration and invasion of breast cancer cells in vitro. These data indicate that Net1 may be a critical regulator of metastatic progression in breast cancer. Net1 activity is negatively regulated by sequestration in the nucleus, and relocalization of Net1 outside the nucleus is required to stimulate RhoA activation, actin cytoskeletal reorganization, and oncogenic transformation. However, regulatory mechanisms controlling the extranuclear localization of Net1 have not been identified. In this study, we have addressed the regulation of Net1A isoform localization by Rac1. Specifically, co-expression of constitutively active Rac1 with Net1A stimulates the relocalization of Net1A from the nucleus to the plasma membrane in breast cancer cells, and results in Net1A activation. Importantly, Net1A localization is also driven by endogenous Rac1 activity. Net1A relocalizes outside the nucleus in cells spreading on collagen, and when endogenous Rac1 expression was silenced by siRNA, Net1A remained nuclear in spreading cells. These data indicate that Rac1 controls the localization of the Net1A isoform and suggests a physiological role for Net1A in breast cancer cell adhesion and motility

A Perl toolkit for LIMS development

BACKGROUND: High throughput laboratory techniques generate huge quantities of scientific data. Laboratory Information Management Systems (LIMS) are a necessary requirement, dealing with sample tracking, data storage and data reporting. Commercial LIMS solutions are available, but these can be both costly and overly complex for the task. The development of bespoke LIMS solutions offers a number of advantages, including the flexibility to fulfil all a laboratory's requirements at a fraction of the price of a commercial system. The programming language Perl is a perfect development solution for LIMS applications because of Perl's powerful but simple to use database and web interaction, it is also well known for enabling rapid application development and deployment, and boasts a very active and helpful developer community. The development of an in house LIMS from scratch however can take considerable time and resources, so programming tools that enable the rapid development of LIMS applications are essential but there are currently no LIMS development tools for Perl. RESULTS: We have developed ArrayPipeline, a Perl toolkit providing object oriented methods that facilitate the rapid development of bespoke LIMS applications. The toolkit includes Perl objects that encapsulate key components of a LIMS, providing methods for creating interactive web pages, interacting with databases, error tracking and reporting, and user and session management. The MT_Plate object provides methods for manipulation and management of microtitre plates, while a given LIMS can be encapsulated by extension of the core modules, providing system specific methods for database interaction and web page management. CONCLUSION: This important addition to the Perl developer's library will make the development of in house LIMS applications quicker and easier encouraging laboratories to create bespoke LIMS applications to meet their specific data management requirements

Ozone profile observations in Houston, Texas (1994 - 2010) from aircraft, balloons, and satellites

Houston, Texas has long been an urban area plagued with high levels of surface ozone, particularly in spring and late summer. The combination of a large commuter population and one of the largest concentrations of petrochemical plants in the world results in abundant and nearly co-located sources of NOx and hydrocarbons. The location of Houston on the South Coast of the United States in a subtropical climate results in meteorological conditions that favor ozone production. Using MOZAIC (1994 - 2004), ozonesonde (2000, 2004 - 2010), and TES (2005 – 2010) data, we examine the evolution of ozone profiles over Houston during a period in which various strategies have been implemented to alleviate the ozone pollution problem. Using meteorological data from associated soundings and analyses, we identify and evaluate influences on the ozone profiles from natural and anthropogenic sources, as well as local and remote sources. We further investigate how these various influences have changed with time

YOGY: a web-based, integrated database to retrieve protein orthologs and associated Gene Ontology terms

We present YOGY a web-based resource for orthologous proteins from nine eukaryotic organisms: Homo sapiens, Mus musculus, Rattus norvegicus, Arabidopsis thaliana, Drosophila melanogaster, Caenorhabditis elegans, Plasmodium falciparum, Schizosaccharomyces pombe and Saccharomyces cerevisiae. Using a gene name from any of these organisms as a query, this database provides comprehensive, combined information on orthologs in other species using data from five independent resources: KOGs, Inparanoid, HomoloGene, OrthoMCL and a table of curated fission and budding yeast orthologs. Associated Gene Ontology (GO) terms of orthologs can also be retrieved for functional inference. Integrating these different and complementary datasets provides a straightforward tool to identify known and predicted orthologs of proteins from a variety of species. This resource should be useful for bench scientists looking for functional clues for their genes of interest as well as for curators looking for information that can be transferred based on orthology and for rapidly identifying the relevant GO terms as an aid to literature curation. YOGY is accessible online at