Functional Analysis of the VirSR Phosphorelay from Clostridium perfringens

Toxin production in Clostridium perfringens is controlled by the VirSR two-component signal transduction system, which comprises the VirS sensor histidine kinase and the VirR response regulator. Other studies have concentrated on the elucidation of the genes controlled by this network; there is little information regarding the phosphorelay cascade that is the hallmark of such regulatory systems. In this study, we have examined each step in this cascade, beginning with autophosphorylation of VirS, followed by phosphotransfer from VirS to VirR. We also have studied the effects of gene dosage and phosphorylation in vivo. We have used random and site-directed mutagenesis to identify residues in VirS that are important for its function and have identified a region in the putative sensory domain of VirS that appeared to be essential for function. In vitro phosphorylation studies showed that VirSc, a truncated VirS protein that lacked the N-terminal sensory domain, was capable of autophosphorylation and could subsequently act as a phosphodonor for its cognate response regulator, VirR. Conserved residues of both VirS and VirR, including the D57 residue of VirR, were shown to be essential for this process. By use of Targetron technology, we were able to introduce a single copy of virR or virRD57N onto the chromosome of a virR mutant of C. perfringens. The results showed that in vivo, when virR was present in single copy, the production of wild-type levels of perfringolysin O was dependent on the presence of virS and an unaltered D57 residue in VirR. These results provide good evidence that phosphorylation is critical for VirR function

Bacterial Toxin Fusion Proteins Elicit Mucosal Immunity against a Foot-and-Mouth Disease Virus Antigen When Administered Intranasally to Guinea Pigs

Peptides corresponding to the foot-and-mouth disease virus VP1 G-H loop are capable of inducing neutralizing antibodies in some species but are considered relatively poor immunogens, especially at mucosal surfaces. However, intranasal administration of antigens along with the appropriate delivery vehicle/adjuvant has been shown to induce mucosal immune responses, and bacterial enterotoxins have long been known to be effective in this regard. In the current study, two different carrier/adjuvant approaches were used to augment mucosal immunity to the FMDV O1 BFS G-H loop epitope, in which the G-H loop was genetically coupled to the E. coli LT-B subunit and coexpressed with the LTA2 fragment (LTA2B-GH), or the nontoxic pseudomonas exotoxin A (ntPE) was fused to LTA2B-GH at LT-A2 to enhance receptor targeting. Only guinea pigs that were inoculated intranasally with ntPE-LTA2B-GH and LTA2B-GH induced significant anti-G-H loop IgA antibodies in nasal washes at weeks 4 and 6 when compared to ovalbumin or G-H loop immunized animals. These were also the only groups that exhibited G-H loop-specific antigen-secreting cells in the nasal mucosa. These data demonstrate that fusion of nonreplicating antigens to LTA2B and ntPE-LTA2B has the potential to be used as carriers/adjuvants to induce mucosal immune responses against infectious diseases

Expression of Human Globin Genes in Transgenic Mice Carrying the Β-Globin Gene Cluster with a Mutation Causing G ΓΒ + Hereditary Persistence of Fetal Hemoglobin a

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73405/1/j.1749-6632.1990.tb24303.x.pd

3-He in the Milky Way Interstellar Medium: Ionization Structure

The cosmic abundance of the 3-He isotope has important implications for many fields of astrophysics. We are using the 8.665 GHz hyperfine transition of 3-He+ to determine the 3-He/H abundance in Milky Way HII regions and planetary nebulae. This is one in a series of papers in which we discuss issues involved in deriving accurate 3-He/H abundance ratios from the available measurements. Here we describe the ionization correction we use to convert the 3-He+/H+ abundance, y3+, to the 3-He/H abundance, y3. In principle the nebular ionization structure can significantly influence the y3 derived for individual sources. We find that in general there is insufficient information available to make a detailed ionization correction. Here we make a simple correction and assess its validity. The correction is based on radio recombination line measurements of H+ and 4-He+, together with simple core-halo source models. We use these models to establish criteria that allow us to identify sources that can be accurately corrected for ionization and those that cannot. We argue that this effect cannot be very large for most of the sources in our observational sample. For a wide range of models of nebular ionization structure we find that the ionization correction factor varies from 1 to 1.8. Although large corrections are possible, there would have to be a conspiracy between the density and ionization structure for us to underestimate the ionization correction by a substantial amount.Comment: 36 pages, 4 figures To appear Astrophysical Journal, 20 August 2007, vol 665, no

The Hubble Space Telescope Key Project on the Extragalactic Distance Scale XXIV: The Calibration of Tully-Fisher Relations and the Value of the Hubble Constant

This paper presents the calibration of BVRIH$ Tully-Fisher relations based on Cepheid distances to 21 galaxies within 25 Mpc, and 23 clusters within 10,000 km/s. These relations have been applied to several distant cluster surveys in order to derive a value for the Hubble constant, H0, mainly concentrating on an I-band all-sky survey by Giovanelli and collaborators which consisted of total I magnitudes and 50% linewidth data for ~550 galaxies in 16 clusters. For comparison, we also derive the values of H0 using surveys in B-band and V-band by Bothun and collaborators, and in H-band by Aaronson and collaborators. Careful comparisons with various other databases from literature suggest that the H-band data, whose magnitudes are isophotal magnitudes extrapolated from aperture magnitudes rather than total magnitudes, are subject to systematic uncertainties. Taking a weighted average of the estimates of Hubble constants from four surveys, we obtain H0 = 71 +- 4 (random) +- 7 (systematic) km/s/Mpc. We have also investigated how various systematic uncertainties affect the value of H0 such as the internal extinction correction method used, Tully-Fisher slopes and shapes, a possible metallicity dependence of the Cepheid period-luminosity relation and cluster population incompleteness bias.Comment: 34 pages, 13 figure

Dynamical age differences among coeval star clusters as revealed by blue stragglers

Globular star clusters that formed at the same cosmic time may have evolved rather differently from a dynamical point of view (because that evolution depends on the internal environment) through a variety of processes that tend progressively to segregate stars more massive than the average towards the cluster centre. Therefore clusters with the same chronological age may have reached quite different stages of their dynamical history (that is, they may have different dynamical ages). Blue straggler stars have masses greater than those at the turn-off point on the main sequence and therefore must be the result of either a collision or a mass-transfer event. Because they are among the most massive and luminous objects in old clusters, they can be used as test particles with which to probe dynamical evolution. Here we report that globular clusters can be grouped into a few distinct families on the basis of the radial distribution of blue stragglers. This grouping corresponds well to an effective ranking of the dynamical stage reached by stellar systems, thereby permitting a direct measure of the cluster dynamical age purely from observed properties.Comment: Published on the 20 December 2012 issue of Natur

GMRT HI observations of the Eridanus group of galaxies

The GMRT HI 21cm-line observations of galaxies in the Eridanus group are presented. The Eridanus group, at a distance of ~23 Mpc, is a loose group of \~200 galaxies. The group extends more than 10 Mpc in projection. The velocity dispersion of the galaxies in the group is ~240 km/s. The galaxies are clustered into different sub-groups. The overall population mix of the group is 30% (E+S0) and 70% (Sp+Irr). The observations of 57 Eridanus galaxies were carried out with the GMRT for ~200 hour. HI emission was detected from 31 galaxies. The channel rms of ~1.0 mJy beam^{-1} was achieved for most of the image-cubes made with 4 hour of data. The corresponding HI column density sensitivity (3-sigma) is ~1x10^{20} cm^{-2} for a velocity-width of ~13.4 km/s. The 3-sigma detection limit of HI mass is ~1.2x10^{7} M_sun for a line-width of 50 km/s. Total HI images, HI velocity fields, global HI line profiles, HI mass surface densities, HI disk parameters and HI rotation curves are presented. The velocity fields are analysed separately for the approaching and the receding sides of the galaxies. This data will be used to study the HI and the radio continuum properties, the Tully-Fisher relations, the dark matter halos, and the kinematical and HI lopsidedness in galaxies.Comment: 75 pages including HI atlas; Accepted for publication in Journal of Astroph. & Astron. March, 200

Dynamics of direct inter-pack encounters in endangered African wild dogs

Aggressive encounters may have important life history consequences due to the potential for injury and death, disease transmission, dispersal opportunities or exclusion from key areas of the home range. Despite this, little is known of their detailed dynamics, mainly due to the difficulties of directly observing encounters in detail. Here, we describe detailed spatial dynamics of inter-pack encounters in African wild dogs (Lycaon pictus), using data from custom-built high-resolution GPS collars in 11 free-ranging packs. On average, each pack encountered another pack approximately every 7 weeks and met each neighbour twice each year. Surprisingly, intruders were more likely to win encounters (winning 78.6% of encounters by remaining closer to the site in the short term). However, intruders did tend to move farther than residents toward their own range core in the short-term (1 h) post-encounter, and if this were used to indicate losing an encounter, then the majority (73.3%) of encounters were won by residents. Surprisingly, relative pack size had little effect on encounter outcome, and injuries were rare (<15% of encounters). These results highlight the difficulty of remotely scoring encounters involving mobile participants away from static defendable food resources. Although inter-pack range overlap was reduced following an encounter, encounter outcome did not seem to drive this, as both packs shifted their ranges post-encounter. Our results indicate that inter-pack encounters may be lower risk than previously suggested and do not appear to influence long-term movement and ranging

HEART score improves efficiency of coronary computed tomography angiography in patients suspected of acute coronary syndrome in the emergency department

Aims: Coronary computed tomography angiography is increasingly employed in the emergency department for suspected acute coronary syndrome patients. The HEART score has been proposed for initial risk stratification in these patients. The aim of this study was to investigate the diagnostic value and efficiency of the HEART score before coronary computed tomography angiography. Methods and results: We included patients suspected of acute coronary syndrome who underwent coronary computed tomography angiography in the emergency department. Based on the HEART score, patients were stratified as low-risk (HEART≤3), intermediate-risk (HEART4–6) and high-risk (HEART≥7). We assessed coronary computed tomography angiography for the presence of significant coronary artery disease (>50% stenosis). The primary outcome, the level of major adverse cardiac events, was a composite endpoint of all-cause mortality, acute coronary syndrome or coronary revascularisation within 30 days. The study population consisted of 340 patients (mean age: 55.6±10.1 years, 44.7% women), major adverse cardiac events occurred in 45 (13.2%) patients. The incidence of major adverse cardiac events in patients stratified as low-risk (35.0%), intermediate-risk (56.8%) and high-risk

Origin and evolution of the light nuclides

After a short historical (and highly subjective) introduction to the field, I discuss our current understanding of the origin and evolution of the light nuclides D, He-3, He-4, Li-6, Li-7, Be-9, B-10 and B-11. Despite considerable observational and theoretical progress, important uncertainties still persist for each and every one of those nuclides. The present-day abundance of D in the local interstellar medium is currently uncertain, making it difficult to infer the recent chemical evolution of the solar neighborhood. To account for the observed quasi-constancy of He-3 abundance from the Big Bang to our days, the stellar production of that nuclide must be negligible; however, the scarce observations of its abundance in planetary nebulae seem to contradict this idea. The observed Be and B evolution as primaries suggests that the source composition of cosmic rays has remained quasi-constant since the early days of the Galaxy, a suggestion with far reaching implications for the origin of cosmic rays; however, the main idea proposed to account for that constancy, namely that superbubbles are at the source of cosmic rays, encounters some serious difficulties. The best explanation for the mismatch between primordial Li and the observed "Spite-plateau" in halo stars appears to be depletion of Li in stellar envelopes, by some yet poorly understood mechanism. But this explanation impacts on the level of the recently discovered early ``Li-6 plateau'', which (if confirmed), seriously challenges current ideas of cosmic ray nucleosynthesis.Comment: 18 pages, 9 figs. Invited Review in "Symposium on the Composition of Matter", honoring Johannes Geiss on the occasion of his 80th birthday (Grindelwald, Switzerland, Sept. 2006), to be published in Space Science Series of ISS