Synthetic chemical inducers and genetic decoupling enable orthogonal control of the rhaBAD promoter

External control of gene expression is crucial in synthetic biology and biotechnology research and applications, and is commonly achieved using inducible promoter systems. The E. coli rhamnose-inducible rhaBAD promoter has properties superior to more commonly-used inducible expression systems, but is marred by transient expression caused by degradation of the native inducer, L-rhamnose. To address this problem, 35 analogs of L-rhamnose were screened for induction of the rhaBAD promoter, but no strong inducers were identified. In the native configuration, an inducer must bind and activate two transcriptional activators, RhaR and RhaS. Therefore, the expression system was reconfigured to decouple the rhaBAD promoter from the native rhaSR regulatory cascade so that candidate inducers need only activate the terminal transcription factor RhaS. Re-screening the 35 compounds using the modified rhaBAD expression system revealed several promising inducers. These were characterised further to determine the strength, kinetics and concentration-dependence of induction; whether the inducer was used as a carbon source by E. coli; and the modality (distribution) of induction among populations of cells. L-Mannose was found to be the most useful orthogonal inducer, providing an even greater range of induction than the native inducer Lrhamnose, and crucially, allowing sustained induction instead of transient induction. These findings address the key limitation of the rhaBAD expression system, and suggest it may now be the most suitable system for many applications

Equations provide tubular information on effects of uniform and variable loads on thin, flat, circular plates

Unit-mass system of derivation of equations determines the deflection, slope, and moments for thin, flat, circular plates subjected to either a uniform or a symmetrical variable load. The derived equations are computed, organized in tabular form, and graphically depicted

IUE archived spectra

The International Ultraviolet Explorer (IUE) Satellite has been in continuous operation since January 26, 1978. To date, approximately 65,000 spectra have been stored in an archive at Goddard Space Flight Center in Greenbelt, MD. A number of procedures have been generated to facilitate access to the data in the IUE spectral archive. This document describes the procedures which include on-line quick look of the displays, search of an observation data base for selected observations, and several methods for ordering data from the archive

Emission Line Galaxies in the STIS Parallel Survey I: Observations and Data Analysis

In the first three years of operation STIS obtained slitless spectra of approximately 2500 fields in parallel to prime HST observations as part of the STIS Parallel Survey (SPS). The archive contains almost 300 fields at high galactic latitude (|b|>30) with spectroscopic exposure times greater than 3000 seconds. This sample contains 220 fields (excluding special regions and requiring a consistent grating angle) observed between 6 June 1997 and 21 September 2000, with a total survey area of about 160 square arcminutes. At this depth, the SPS detects an average of one emission line galaxy per three fields. We present the analysis of these data, and the identification of 131 low to intermediate redshift galaxies detected by optical emission lines. The sample contains 78 objects with emission lines that we infer to be redshifted [OII]3727 emission at 0.43<z<1.7. The comoving number density of these objects is comparable to that of H-alpha emitting galaxies in the NICMOS parallel observations. One quasar and three probable Seyfert galaxies are detected. Many of the emission-line objects show morphologies suggestive of mergers or interactions. The reduced data are available upon request from the authors.Comment: 58 preprint pages, including 26 figures; accepted for publication in ApJ

A Candidate Substellar Companion to HR 7329

We present the discovery of a candidate substellar companion from a survey of nearby, young stars with the NICMOS coronagraph on the Hubble Space Telescope. The H ~ 12 mag object was discovered approximately 4" from the young A0V star HR 7329. Using follow-up spectroscopy from STIS, we derive a spectral type between M7V and M8V with an effective temperature of ~ 2600 K. We estimate that the probability of a chance alignment with a foreground dwarf star of this nature is ~ 10^(-8) and therefore suggest the object (HR 7329B) is physically associated with HR 7329 with a projected separation of 200 AU. Current brown dwarf cooling models indicate a mass of less than 50 Jupiter masses for HR 7329B based on age estimates of < 30 Myr for HR7329A.Comment: 8 pages LATEX, 5 ps figures, accepted for Ap

The Heavy Element Enrichment of Lyman alpha Clouds in the Virgo Supercluster

Using high S/N STIS echelle spectra (FWHM=7 km/s) of 3C 273, we constrain the metallicities of two Lya clouds in the vicinity of the Virgo cluster. We detect C II, Si II, and Si III absorption lines in the Lya absorber at z = 0.00530. Previous observations with FUSE have revealed Ly beta - Ly theta lines at this redshift, thereby accurately constraining N(H I). We model the ionization of the gas and derive [C/H] = -1.2^{+0.3}_{-0.2}, [Si/C] = 0.2+/-0.1, and log n_{H} = -2.8+/-0.3. The model implies a small absorber thickness, ~70 pc, and thermal pressure p/k ~ 40 cm^{-3} K. It is most likely that the absorber is pressure confined by an external medium because gravitational confinement would require a very high ratio of dark matter to baryonic matter. Based on Milky Way sight lines in which carbon and silicon abundances have been reliably measured in the same interstellar cloud (including new measurements presented herein), we argue that the overabundance of Si relative to C is not due to dust depletion. Instead, this probably indicates that the gas has been predominately enriched by Type II supernovae. Such enrichment is most plausibly provided by an unbound galactic wind, given the absence of galaxies within a projected distance of 100 kpc and the presence of galaxies capable of driving a wind at larger distances. We also constrain the metallicity and physical conditions of the Virgo absorber at z = 0.00337 based on detections of O VI and H I and an upper limit on C IV. If this absorber is collisionally ionized, the O VI/C IV limit requires T > 10^{5.3} K. For either collisional ionization or photoionization, we find that [O/H] > -2.0 at z = 0.00337.Comment: Final Ap.J. versio

Paraphrases and summaries: A means of clarification or a vehicle for articulating a preferred version of student accounts?

The use of group discussions as a means to facilitate learning from experiences is well documented in adventure education literature. Priest and Naismith (1993) assert that the use of the circular discussion method, where the leader poses questions to the participants, is the most common form of facilitation in adventure education. This paper draws on transcripts of facilitation sessions to argue that the widely advocated practice of leader summaries or paraphrases of student responses in these sessions functions as a potential mechanism to control and sponsor particular knowledge(s). Using transcripts from recorded facilitation sessions the analysis focuses on how the leader paraphrases the students’ responses and how these paraphrases or ‘formulations’ function to modify or exclude particular aspects of the students’ responses. I assert that paraphrasing is not simply a neutral activity that merely functions to clarify a student response, it is a subtle means by which the leader of the session can, often inadvertently or unknowingly, alter the student’s reply with the consequence of favouring particular knowledge(s). Revealing the subtle work that leader paraphrases perform is of importance for educators who claim to provide genuine opportunities for students to learn from their experience

Complex nature of SNP genotype effects on gene expression in primary human leucocytes.

This is a freely-available open access publication. Please cite the published version which is available via the DOI link in this record.BACKGROUND: Genome wide association studies have been hugely successful in identifying disease risk variants, yet most variants do not lead to coding changes and how variants influence biological function is usually unknown. METHODS: We correlated gene expression and genetic variation in untouched primary leucocytes (n = 110) from individuals with celiac disease - a common condition with multiple risk variants identified. We compared our observations with an EBV-transformed HapMap B cell line dataset (n = 90), and performed a meta-analysis to increase power to detect non-tissue specific effects. RESULTS: In celiac peripheral blood, 2,315 SNP variants influenced gene expression at 765 different transcripts (< 250 kb from SNP, at FDR = 0.05, cis expression quantitative trait loci, eQTLs). 135 of the detected SNP-probe effects (reflecting 51 unique probes) were also detected in a HapMap B cell line published dataset, all with effects in the same allelic direction. Overall gene expression differences within the two datasets predominantly explain the limited overlap in observed cis-eQTLs. Celiac associated risk variants from two regions, containing genes IL18RAP and CCR3, showed significant cis genotype-expression correlations in the peripheral blood but not in the B cell line datasets. We identified 14 genes where a SNP affected the expression of different probes within the same gene, but in opposite allelic directions. By incorporating genetic variation in co-expression analyses, functional relationships between genes can be more significantly detected. CONCLUSION: In conclusion, the complex nature of genotypic effects in human populations makes the use of a relevant tissue, large datasets, and analysis of different exons essential to enable the identification of the function for many genetic risk variants in common diseases.Coeliac UKNetherlands Organization for Scientific ResearchCeliac Disease Consortium (an innovative cluster approved by the Netherlands Genomics Initiative and partly funded by the Dutch government)Netherlands Genomics InitiativeWellcome Trus

Terminal velocities of luminous, early-type SMC stars

Ultraviolet spectra from the Space Telescope Imaging Spectrograph (STIS) are used to determine terminal velocities for 11 O and B-type giants and supergiants in the Small Magellanic Cloud (SMC) from the Si IV and C IV resonance lines. Using archival data from observations with the Goddard High-Resolution Spectrograph and the International Ultraviolet Explorer telescope, terminal velocities are obtained for a further five B-type supergiants. We discuss the metallicity dependence of stellar terminal velocities, finding no evidence for a significant scaling between Galactic and SMC metallicities for Teff < 30,000 K, consistent with the predictions of radiation driven wind theory for supergiant stars. A comparison of the $v_\infty / v_{esc}$ ratio between the SMC and Galactic samples, while consistent with the above statement, emphasizes that the uncertainties in the distances to galactic O-stars are a serious obstacle to a detailed comparison with theory. For the SMC sample there is considerable scatter in this ratio at a given effective temperature, perhaps indicative of uncertainties in stellar masses.Comment: 28 pages, 8 figures, accepted by ApJ; minor revisions prior to acceptanc