Directed evolution of Vibrio fischeri LuxR for increased sensitivity to a broad spectrum of acyl-homoserine lactones

LuxR-type transcriptional regulators play key roles in quorum-sensing systems that employ acyl-homoserine lactones (acyl-HSLs) as signal molecules. These proteins mediate quorum control by changing their interactions with RNA polymerase and DNA in response to binding their cognate acyl-HSL. The evolutionarily related LuxR-type proteins exhibit considerable diversity in primary sequence and in their response to acyl-HSLs having acyl groups of differing length and composition. Little is known about which residues determine acyl-HSL specificity, and less about the evolutionary time scales required to forge new ones. To begin to examine such issues, we have focused on the LuxR protein from Vibrio fischeri, which activates gene transcription in response to binding its cognate quorum signal, 3-oxohexanoyl-homoserine lactone (3OC6HSL). Libraries of luxR mutants were screened for variants exhibiting increased gene activation in response to octanoyl-HSL (C8HSL), with which wild-type LuxR interacts only weakly. Eight LuxR variants were identified that showed a 100-fold increase in sensitivity to C8HSL; these variants also displayed increased sensitivities to pentanoyl-HSL and tetradecanoyl-HSL, while maintaining a wild-type or greater response to 3OC6HSL. The most sensitive variants activated gene transcription as strongly with C8HSL as the wild type did with 3OC6HSL. With one exception, the amino acid residues involved were restricted to the N-terminal, 'signal-binding' domain of LuxR. These residue positions differed from critical positions previously identified via 'loss-of-function' mutagenesis. We have demonstrated that acyl-HSL-dependent quorum-sensing systems can evolve rapidly to respond to new acyl-HSLs, suggesting that there may be an evolutionary advantage to maintaining such plasticity

Test instrumentation evaluates electrostatic hazards in fluid system

RJ-1 fuel surface potential is measured with a probe to determine the degree of hazard originating from static electricity buildup in the hydraulic fluid. The probe is mounted in contact with the fluid surface and connected to an electrostatic voltmeter

The application of amino acid racemization in the acid soluble fraction of enamel to the estimation of the age of human teeth

Estimation of age-at-death for skeletonised forensic remains is one of the most significant problems in forensic anthropology. The majority of existing morphological and histological techniques are highly inaccurate, and show a bias towards underestimating the age of older individuals. One technique which has been successful in forensic age estimation is amino acid racemization in dentine. However, this method cannot be used on remains where the post-mortem interval is greater than 20 years. An alternative approach is to measure amino acid racemization in dental enamel, which is believed to be more resistant to change post-mortem. The extent of amino acid racemization in the acid soluble fraction of the enamel proteins was determined for modem known age teeth. A strong correlation was observed between the age of the tooth and the extent of racemization. No systematic bias in the direction of age estimation errors was detected. For the majority of teeth analyzed, the presence of dental caries did not affect the results obtained. In a minority of cases, carious teeth showed a higher level of racemization than would be expected given the age of the individual. These results indicate that amino acid racemization in enamel has the potential to be used in age estimation of skeletal remains. (C) 2007 Elsevier Ireland Ltd. All rights reserved

State Trends in Premiums and Deductibles, 2003-2009: How Building on the Affordable Care Act Will Help Stem the Tide of Rising Costs and Eroding Benefits

Examines 2003-09 state trends in family coverage premiums and deductibles for private employers. Looks at projected savings on premiums if the 2010 healthcare reform succeeds in slowing growth by 1 percentage point annually and weighs policy implications

Experience and Assessment of the DOE/NASA Mod-1 2000 Kw Wind Turbine Generator at Boone, North Carolina

The Mod 1 program objectives are defined. The Mod 1 wind turbine is described. In addition to the steel blade operated on the wind turbine, a composite blade was designed and manufactured. During the early phase of the manufacturing cycle of Mod 1A configuration was designed that identified concepts such as partial span control, a soft tower, and upwind teetered rotors that were incorporated in second and third generation industry designs. The Mod 1 electrical system performed as designed, with voltage flicker characteristics within acceptable utility limits

Type II superlattices for infrared detectors and devices

Superlattices consisting of combinations of III-V semiconductors with type II band alignments are of interest for infrared applications because their energy gaps can be made smaller than those of any 'natural' III-V compounds. Specifically, it has been demonstrated that both InSb/InAsxSb1-x superlattices and Ga1-xInxSb/InAs superlattices can possess energy gaps in the 8-14 mu m range. The efforts have focused on the Ga1-xInxSb/InAs system because of its extreme broken gap band alignment, which results in narrow energy gaps for very short superlattice periods. The authors report the use of in situ chemical doping of Ga1-xInxSb/InAs superlattices to fabricate p-n photodiodes. These diodes display a clear photovoltaic response with a threshold near 12 mu m. They have also attained outstanding structural quality in Ga1-xInxSb/InAs superlattices grown on radiatively heated GaSb substrates. Cross-sectional transmission electron microscope images of these superlattices display no dislocations, while high resolution X-ray diffraction scans reveal sharp high-order superlattice satellites and strong Pendellosung fringes