Development of real-time PCR and hybridization methods for detection and identification of thermophilic Campylobacter spp. in pig faecal samples

Aims: To develop a real-time (rt) PCR for species differentiation of thermophilic Campylobacter and to develop a method for assessing co-colonization of pigs by Campylobacter spp. Methods and results: The specificity of a developed 5’nuclease rt-PCR for species-specific identification of C. jejuni, C. coli, C. lari, C. upsaliensis and of a hipO gene nucleotide probe for detection of C. jejuni by colony-blot hybridization were determined by testing a total of 75 reference strains of Campylobacter spp. and related organisms. The rt-PCR method allowed species-specific detection of Campylobacter spp. in naturally infected pig faecal samples after an enrichment step, whereas the hybridization approach enhanced the specific isolation of C. jejuni (present in minority to C. coli) from pigs. Conclusions: The rt-PCR was specific for Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis and the colony-blot hybridization approach provided an effective tool for isolation of C. jejuni from pig faecal samples typically dominated by C. coli. Significance and impact of study: Species differentiation between thermophilic Campylobacter is difficult by phenotypic methods and the developed rt-PCR provides an easy and fast method for such differentiation. Detection of C. jejuni by colony hybridization may increase the isolation rate of this species from pig feces

Great Circle tidal streams: evidence for a nearly spherical massive dark halo around the Milky Way

An all high-latitude sky survey for cool carbon giant stars in the Galactic halo has revealed 75 such stars, of which the majority are new detections. Of these, more than half are clustered on a Great Circle on the sky which intersects the center of Sagittarius dwarf galaxy (Sgr) and is parallel to its proper motion vector, while many of the remainder are outlying Magellanic Cloud C-stars. A pole-count analysis of the carbon star distribution clearly indicates that the Great Circle stream we have isolated is statistically significant, being a 5-6 sigma over-density. These two arguments strongly support our conclusion that a large fraction of the Halo carbon stars originated in Sgr. The stream orbits the Galaxy between the present location of Sgr, 16 kpc from the Galactic center, and the most distant stream carbon star, at ~60 kpc. It follows neither a polar nor a Galactic plane orbit, so that a large range in both Galactic R and z distances are probed. That the stream is observed as a Great Circle indicates that the Galaxy does not exert a significant torque upon the stream, so the Galactic potential must be nearly spherical in the regions probed by the stream. We present N-body experiments simulating this disruption process as a function of the distribution of mass in the Galactic halo. A likelihood analysis shows that, in the Galactocentric distance range 16 kpc < R < 60 kpc, the dark halo is most likely almost spherical. We rule out, at high confidence levels, the possibility that the Halo is significantly oblate, with isodensity contours of aspect q_m < 0.7. This result is quite unexpected and contests currently popular galaxy formation models. (Abridged)Comment: 26 pages, 13 figures (6 in color, 8 chunky due to PS compression), minor revisions, accepted by Ap

Intranasal nanoemulsion vaccine confers long‐lasting immunomodulation and sustained unresponsiveness in a murine model of milk allergy

BackgroundImmunotherapy for food allergy requires prolonged treatment protocols and, in most cases, does not lead to durable modulation of the allergic immune response. We have demonstrated an intranasal (IN) nanoemulsion adjuvant that redirects allergen‐specific Th2 responses toward Th1 and Th17 immunity, and protects from allergen challenge after only 2‐4 monthly administrations. Here, we investigate the ability of this technology to provide long‐term modulation of allergy in a murine model of cow’s milk allergy.MethodsSix weeks after sensitization to bovine casein, mice received four, monthly IN immunizations with nanoemulsion formulated with casein. Protection from casein challenge was assessed at 4 and 16 weeks after the final vaccine administration.ResultsThe NE vaccine significantly blunted the physiological responses to allergen challenge, and this effect persisted for at least 16 weeks. The protection from challenge was associated with the suppression of casein‐specific Th2 immunity and induced Th1 and Th17 cytokines as well as induction of IL‐10. Of interest, while immunized animals showed significantly decreased Th2 cytokine responses, cow’s milk‐specific IgE remained elevated in the serum at levels associated with reactivity in control sensitized animals. Protection was associated with suppressed mast cell activation and markedly reduced mast cell infiltration into the small intestine.ConclusionThe sustained unresponsiveness of at least 16 weeks after vaccination suggests that the nanoemulsion vaccine alters the allergic phenotype in a persistent manner different from traditional desensitization, and this leads to long‐term suppressive effects on allergic disease without eliminating serum IgE.This study evaluates the ability of an intranasal nanoemulsion‐based vaccine to induce long‐term modulation of allergic reactions in a mouse model of cow’s milk allergy. Intranasal immunization with nanoemulsion adjuvant suppresses Th2 responses and anaphylaxis. The sustained unresponsiveness of at least 16 weeks after vaccination suggests that the nanoemulsion vaccine alters the allergic phenotype.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154959/1/all14064_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/154959/2/all14064.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/154959/3/all14064-sup-0003-FigS3.pd

NICMOS and VLA Observations of the Gravitatonally Lensed Ultraluminous BAL Quasar APM~08279+5255: Detection of a Third Image

We present a suite of observations of the recently identified ultraluminous BAL quasar APM 08279+5255, taken both in the infra-red with the NICMOS high resolution camera on board the Hubble Space Telescope, and at 3.5cm with the Very Large Array. With an inferred luminosity of ~5x10^15 Solar luminosities, APM 08279+5255 is apparently the most luminous system known. Extant ground-based images show that APM 08279+5255 is not point-like, but is instead separated into two components, indicative of gravitational lensing. The much higher resolution images presented here also reveal two point sources, A and B, of almost equal brightness (f_B/f_A=0.782 +/- 0.010), separated by 0."378 +/- 0."001, as well as a third, previously unknown, fainter image, C, seen between the brighter images. While the nature of C is not fully determined, several lines of evidence point to it being a third gravitationally lensed image of the quasar, rather than being the lensing galaxy. Simple models which recover the relative image configuration and brightnesses are presented. While proving to be substantially amplified, APM 08279+5255 possesses an intrinsic bolometric luminosity of ~10^14 to 10^15 Solar luminosities and remains amongst the most luminous objects known.Comment: 21 pages, 5 figures (2 as GIF files); accepted for publication in the Astronomical Journa

A Process and Outcome Evaluation of a Shelter for Homeless Young Women

To evaluate the processes and outcomes of a short-term shelter, both quantitative and qualitative data were gathered via participant observation, focus group interviews with shelter staff and residents, and individual interviews with a sample of 40 young women who had been homeless prior to using the shelter. The process evaluation showed that the shelter staff strived to utilize an empowerment philosophy in their relationships with residents, but that there were many challenges to implementing this philosophy. The outcome evaluation showed that, at a 3-month follow-up, the participants reported significant improvements in housing, income, independence, and life satisfaction, but most continued to experience poverty and a number of other difficulties. The results were discussed in terms of the implications for future research and the value and limitations of shelters for dealing with homeless youth. The need for more sustained and comprehensive program interventions and supportive social policies was underscored

X-ray Source Populations in the Region of the Open Clusters NGC 6633 and IC 4756

Using deep exposures (~10^5 s) with the ROSAT HRI, we have performed flux-limited surveys for X-ray sources in the vicinity of the Hyades-age open stellar clusters NGC 6633 and IC 4756, detecting 31 and 13 sources respectively. Our primary aim is to search for so-far unrecognised cluster members. We propose identifications or classifications (cluster member, field star, extragalactic field object) for the X-ray sources, based on published membership lists, and on X-ray:optical flux ratios and optical colour--magnitude diagrams. Results of simulating the expected X-ray-emitting source populations are compared with the ROSAT measurements and with the expected capabilities of the XMM mission. The simulations provide a novel method of comparing the activity levels of NGC 6633 and IC 4756 with that of the Hyades. The measurements and simulations confirm that cluster members are the major class of X-ray emitter in these fields at flux levels f_x > 10^{-14}/erg/cm^2/s (0.1--2.4 keV), contributing ~40 per cent of the total X-ray sources. We find 6 possible new members in NGC 6633, and 4 candidates in IC 4756; all require further observation to establish membership probability.Comment: 12 pages, 4 figure

Identifying risks for male street gang affiliation: a systematic review and narrative synthesis

Gang violence has increased in recent years. Individuals are becoming gang affiliated younger, and many have suffered historic maltreatment. Subsequent exposure to violence can result in profound consequences, including acute psychological harm. This review aims to identify predictive risk factors for male street gang affiliation. A systematic literature search was conducted utilising PsycINFO, PsycARTICLES, Medline, the Cochrane Central Register of Controlled Trials, the Cochrane Database of Systematic Reviews and the Social Policy and Practice databases (from the databases’ inception to 03/04/15). From this search, n=244 peer-reviewed papers were included in an initial scoping review, and n=102 thereafter met criteria for a systematic review; a narrative synthesis follows. Gang members have typically faced numerous historic adversities across multiple domains; individual, family, peers, school and community. Cumulative factors generated an independent risk. The meta-narrative described an overarching failure to safeguard vulnerable individuals, with the motivation for gang affiliation hypothetically arising from an attempt to have their basic needs met. Clinical and research recommendations were made to inform early intervention policy and practice

Comparative genomics of isolates of a pseudomonas aeruginosa epidemic strain associated with chronic lung infections of cystic fibrosis patients

Pseudomonas aeruginosa is the main cause of fatal chronic lung infections among individuals suffering from cystic fibrosis (CF). During the past 15 years, particularly aggressive strains transmitted among CF patients have been identified, initially in Europe and more recently in Canada. The aim of this study was to generate high-quality genome sequences for 7 isolates of the Liverpool epidemic strain (LES) from the United Kingdom and Canada representing different virulence characteristics in order to: (1) associate comparative genomics results with virulence factor variability and (2) identify genomic and/or phenotypic divergence between the two geographical locations. We performed phenotypic characterization of pyoverdine, pyocyanin, motility, biofilm formation, and proteolytic activity. We also assessed the degree of virulence using the Dictyostelium discoideum amoeba model. Comparative genomics analysis revealed at least one large deletion (40-50 kb) in 6 out of the 7 isolates compared to the reference genome of LESB58. These deletions correspond to prophages, which are known to increase the competitiveness of LESB58 in chronic lung infection. We also identified 308 non-synonymous polymorphisms, of which 28 were associated with virulence determinants and 52 with regulatory proteins. At the phenotypic level, isolates showed extensive variability in production of pyocyanin, pyoverdine, proteases and biofilm as well as in swimming motility, while being predominantly avirulent in the amoeba model. Isolates from the two continents were phylogenetically and phenotypically undistinguishable. Most regulatory mutations were isolate-specific and 29% of them were predicted to have high functional impact. Therefore, polymorphism in regulatory genes is likely to be an important basis for phenotypic diversity among LES isolates, which in turn might contribute to this strain's adaptability to varying conditions in the CF lung

RECENT RESULTS FOR THE DEPENDENCE OF BEAM INSTABILITIES CAUSED BY ELECTRON CLOUDS AT CESRTA DUE TO VARIATIONS IN BUNCH SPACING AND CHROMATICITY*

Abstract At the Cornell Electron-Positron Storage Ring Test Accelerator (CesrTA) experiments have been studying the interaction of the electron cloud (EC) with 2.1 GeV stored electron and positron beams. These experiments are intended to characterize the dependence of beam-EC interactions on various beam parameters, such as bunch spacing and vertical chromaticity. Most experiments were performed with 30 or 45-bunch trains, at a fixed current of 0.75 mA/bunch. Earlier experiments with positrons had varied the bunch spacing between 4 and 56 ns at three different vertical chromaticity settings. More recent measurements have included electron-bunch trains to contrast the build up of EC between electron and positron beams. The dynamics of the stored beam was quantified using: a gated Beam Position Monitor (BPM) and spectrum analyzer to measure the bunch-by-bunch frequency spectrum of the bunch trains; an x-ray beam size monitor to record the bunch-by-bunch, turn-by-turn vertical size of each bunch within the trains. We report on recent observations from these experiments and additional studies, using witness bunches trailing 30 or 45-bunch positron trains, which were used for the generation of the ECs

Optical spectroscopy of the M15 intermediate velocity cloud

We present echelle spectrograph observations in NaD, at resolutions of 6.2-8.5 km/s, for 11 stars located in the line-of-sight to the M15 intermediate velocity cloud, which has a radial velocity of 70 km/s. This cloud is a part of IVC Complex gp. The targets range in magnitude from V=13.3-14.8. Seven of the observed stars are in the M15 globular cluster, the remaining four being field stars. Column density ratios of log(N cm^-2)=11.8-12.5 are derived. Combining the current sightlines with previously-existing data, we find the NaD/HI ratio in the IVC varies by upto a factor of 25. One cluster star, M15 ZNG-1, was also observed in Calcium. We find N(CaI)/N(CaII)<0.03 and NaI/CaII=0.25, similar to values seen in the local ISM. Finally, we detect tentative evidence for IV absorption in KI towards 3 cluster stars.Comment: A&A, in pres