Long-distance communication in synthetic bacterial consortia through active signal propagation

A synthetic cell-cell signaling circuit should ideally be (1) metabolically lightweight, (2) insulated from endogenous gene networks, and (3) excitable rather than oscillatory or bistable. To accomplish these three features, we propose a synchronized pulse-generating circuit based on the design of published synchronized oscillators. This communication module employs a pulse generator built using Lux-type quorum sensing components and an IFFL transcriptional circuit. Both the input and output of this module are AHLs, the quorum sensing signaling molecule. Cells bearing this module therefore act as an excitable medium, producing a pulse of AHL when stimulated by exogenous AHL. Using simulation and microscopy, we demonstrate how this circuit enables traveling pulses of AHL production through microcolonies growing in two dimensions. Traveling pulses achieve cell-cell communication at longer distances than can be achieved by diffusion of signal from sender to receiver cells and may permit more sophisticated coordination in synthetic consortia

Timeless Strategy Meets New Medium: Going Negative on Congressional Campaign Websites, 2002-2006

Abstract: In a few short years, the World Wide Web has become a standard part of candidates’ campaign tool kits. Virtually all candidates have their own sites; and voters, journalists, and activists visit the sites with increasing frequency. In this paper, we study what candidates do on these sites—in terms of the information they present—by exploring one of the most enduring and widely debated campaign strategies: “going negative. ” Comparing data from over 700 congressional candidate websites, over three election cycles (2002, 2004, and 2006), with television advertising data, we show that candidates go negative with similar likelihoods across these media. We also find that while similar dynamics drive negativity on the Web and in television advertising, there are some notable differences. These differences likely stem, in part, from the truncated sample available with television data (i.e., many candidates do not produce ads). Our results have implications for understanding negative campaigning, and for the ways in which scholars can study campaign dynamics

Length and time scales of cell-cell signaling circuits in agar

A community of genetically heterogeneous cells embedded in an unmixed medium allows for sophisticated operations by retaining spatial differentiation and coordinating division-of-labor. To establish the principles of engineering reliable cell-cell communication in a heterogeneous environment, we examined how circuit parameters and spatial placement affect the range of length and time scales over which simple communication circuits interact. We constructed several "sender" and "receiver" strains with quorum-sensing signaling circuits. The sender cell colony produces acyl homoserine lactones (AHL), which diffuse across the semisolid medium. The receiver cell colony detects these signal molecules and reports by fluorescence. We have found that a single colony of one sender variant is sufficient to induce receiver response at more than 1.5cm separation. Furthermore, AHL degradase expression in receiver colonies produces a signal threshold effect and reduces the response level in subsequent receiver colonies. Finally, our investigation on the spatial placement of colonies gave rise to the design of a multicellular long-range communication array consisting of two alternating colony types. Its signal response successfully propagated colony-by-colony along a six-colony array spanning 4.8cm at a transmission velocity of 12.8 hours per colony or 0.075cm per hour. In addition, we have developed a reaction-diffusion model that recreates the observed behaviors of the many performed experiments using data-informed parameter estimates of signal diffusion, gene expression, and nutrient consumption. These results demonstrate that a mixed community of colonies can enable new patterning programs, and the corresponding model will facilitate the rational design of complex communication networks

Engineering pulsatile communication in bacterial consortia

Lux-type quorum sensing systems enable communication in bacteria with only two protein components: a signal synthase and an inducible transcription activator. The simplicity of these systems makes them a popular choice for engineering collaborative behaviors in synthetic bacterial consortia, such as photographic edge detection and synchronized oscillation. To add to this body of work, we propose a pulsatile communication circuit that enables dynamic patterning and long-distance communication analogous to action potentials traveling through nerve tissue. We employed a model-driven design paradigm involving incremental characterization of in vivo design candidates with increasing circuit complexity. Beginning with a simple inducible reporter system, we screened a small number of circuits varying in their promoter and ribosomal binding site strengths. From this candidate pool, we selected a candidate to be the seed network for the subsequent round of more complex circuit variants, likewise variable in promoter and RBS strengths. The selection criteria at each level of complexity is tailored to optimize a different desirable performance characteristic. By this approach we individually optimized reporter signal-to-background ratio, pulsatile response to induction, and quiescent basal transcription, avoiding large library screens while ensuring robust performance of the composite circuit

National Educators' Workshop: Update 1996

This document contains a collection of experiments presented and demonstrated at the National Educators' Workshop: Update 96, held at Los Alamos National Laboratory, Los Alamos, New Mexico on October 27-30, 1996. The experiments related to the nature and properties of engineering materials and provided information to assist in teaching about materials in the education community

Development and Short-Range Testing of a 100 kW Side-Illuminated Millimeter-Wave Thermal Rocket

The objective of the phase described here of the Millimeter-Wave Thermal Launch System (MTLS) Project was to launch a small thermal rocket into the air using millimeter waves. The preliminary results of the first MTLS flight vehicle launches are presented in this work. The design and construction of a small thermal rocket with a planar ceramic heat exchanger mounted along the axis of the rocket is described. The heat exchanger was illuminated from the side by a millimeter-wave beam and fed propellant from above via a small tank containing high pressure argon or nitrogen. Short-range tests where the rocket was launched, tracked, and heated with the beam are described. The rockets were approximately 1.5 meters in length and 65 millimeters in diameter, with a liftoff mass of 1.8 kilograms. The rocket airframes were coated in aluminum and had a parachute recovery system activated via a timer and Pyrodex. At the rocket heat exchanger, the beam distance was 40 meters with a peak power intensity of 77 watts per square centimeter. and a total power of 32 kilowatts in a 30 centimeter diameter circle. An altitude of approximately 10 meters was achieved. Recommendations for improvements are discussed

Re-engineering a NiFe hydrogenase to increase the H2 production bias while maintaining native levels of O2 tolerance

Naturally occurring oxygen tolerant NiFe membrane bound hydrogenases have a conserved catalytic bias towards hydrogen oxidation which limits their technological value. We present an Escherichia coli Hyd-1 amino acid exchange that apparently causes the catalytic rate of H2 production to double but does not impact the O2 tolerance

Long-distance communication in synthetic bacterial consortia through active signal propagation

A synthetic cell-cell signaling circuit should ideally be (1) metabolically lightweight, (2) insulated from endogenous gene networks, and (3) excitable rather than oscillatory or bistable. To accomplish these three features, we propose a synchronized pulse-generating circuit based on the design of published synchronized oscillators. This communication module employs a pulse generator built using Lux-type quorum sensing components and an IFFL transcriptional circuit. Both the input and output of this module are AHLs, the quorum sensing signaling molecule. Cells bearing this module therefore act as an excitable medium, producing a pulse of AHL when stimulated by exogenous AHL. Using simulation and microscopy, we demonstrate how this circuit enables traveling pulses of AHL production through microcolonies growing in two dimensions. Traveling pulses achieve cell-cell communication at longer distances than can be achieved by diffusion of signal from sender to receiver cells and may permit more sophisticated coordination in synthetic consortia