Immuno-analysis of microparticles: Probing at the limits of detection

Microparticle (MP) research is clouded by debate regarding the accuracy and validity of flow cytometry (FCM) as an analytical methodology, as it is influenced by many variables including the pre-analytical conditions, instruments physical capabilities and detection parameters. This study utilises a simplistic in vitro system for generating MP, and through comparative analysis with immuno-electron microscopy (Immuno-EM) assesses the strengths and limitations of probe selection and high-sensitivity FCM. Of the markers examined, MP were most specifically labelled with phosphatidylserine ligands, annexin V and lactadherin, although only ∼60% MP are PS positive. Whilst these two ligands detect comparable absolute MP numbers, they interact with the same population in distinct manners; annexin V binding is enhanced on TNF induced MP. CD105 and CD54 expression were, as expected, consistent and enhanced following TNF activation respectively. Their labelling however accounted for as few as 30-40% of MP. The greatest discrepancies between FCM and I-EM were observed in the population solely labelled for the surface antigen. These findings demonstrate that despite significant improvements in resolution, high-sensitivity FCM remains limited in detecting small-size MP expressing low antigen levels. This study highlights factors to consider when selecting endothelial MP probes, as well as interpreting and representing data

Quasars in the MAMBO blank field survey

Our MAMBO 1.2 mm blank field imaging survey of ~0.75 sqd has uncovered four unusually bright sources, with flux densities between 10 and 90 mJy, all located in the Abell 2125 field. The three brightest are flat spectrum radio sources with bright optical and X-ray counterparts. Their mm and radio flux densities are variable on timescales of months. Their X-ray luminosities classify them as quasars. The faintest of the four mm bright sources appears to be a bright, radio-quiet starburst at z~3, similar to the sources seen at lower flux densities in the MAMBO and SCUBA surveys. It may also host a mildly obscured AGN of quasar-like X-ray luminosity. The three non-thermal mm sources imply an areal density of flat spectrum radio sources higher by at least 7 compared with that expected from an extrapolation of the lower frequency radio number counts.Comment: 8 pages, 7 figures. Accepted for publication by A&

Timing and conditions of peak metamorphism and cooling across the Zimithang Thrust, Arunachal Pradesh, India

The Zimithang Thrust juxtaposes two lithotectonic units of the Greater Himalayan Sequence in Arunachal Pradesh, NE India. Monazite U–Pb, muscovite 40Ar/39Ar and thermobarometric data from rocks in the hanging and footwall constrain the timing and conditions of their juxtaposition across the structure, and their subsequent cooling. Monazite grains in biotite–sillimanite gneiss in the hanging wall yield LA-ICP-MS U–Pb ages of 16 ± 0.2 to 12.7 ± 0.4 Ma. A schistose gneiss within the high strain zone yields overlapping-to-younger monazite ages of 14.9 ± 0.3 to 11.5 ± 0.3 Ma. Garnet–staurolite–mica schists in the immediate footwall yield older monazite ages of 27.3 ± 0.6 to 17.1 ± 0.2 Ma. Temperature estimates from Ti-in-biotite and garnet–biotite thermometry suggest similar peak temperatures were achieved in the hanging and footwalls (~ 525–650 °C). Elevated temperatures of ~ 700 °C appear to have been reached in the high strain zone itself and in the footwall further from the thrust. Single grain fusion 40Ar/39Ar muscovite data from samples either side of the thrust yield ages of ~ 7 Ma, suggesting that movement along the thrust juxtaposed the two units by the time the closure temperature of Ar diffusion in muscovite had been reached. These data confirm previous suggestions that major orogen-parallel out-of-sequence structures disrupt the Greater Himalayan Sequence at different times during Himalayan evolution, and highlight an eastwards-younging trend in 40Ar/39Ar muscovite cooling ages at equivalent structural levels along Himalayan strike

Genomic surveillance of Escherichia coli and Klebsiella spp. in hospital sink drains and patients

Escherichia coli and Klebsiella spp. are important human pathogens that cause a wide spectrum of clinical disease. In healthcare settings, sinks and other wastewater sites have been shown to be reservoirs of antimicrobial-resistant E. coli and Klebsiella spp., particularly in the context of outbreaks of resistant strains amongst patients. Without focusing exclusively on resistance markers or a clinical outbreak, we demonstrate that many hospital sink drains are abundantly and persistently colonised with diverse populations of E. coli, Klebsiella pneumoniae and Klebsiella oxytoca, including both antimicrobial-resistant and susceptible strains. Using whole genome sequencing (WGS) of 439 isolates, we show that environmental bacterial populations are largely structured by ward and sink, with only a handful of lineages, such as E. coli ST635, being widely distributed, suggesting different prevailing ecologies which may vary as a result of different inputs and selection pressures. WGS of 46 contemporaneous patient isolates identified one (2%; 95% CI 0.05-11%) E. coli urine infection-associated isolate with high similarity to a prior sink isolate, suggesting that sinks may contribute to up to 10% of infections caused by these organisms in patients on the ward over the same timeframe. Using metagenomics from 20 sink-timepoints, we show that sinks also harbour many clinically relevant antimicrobial resistance genes including blaCTX-M, blaSHV and mcr, and may act as niches for the exchange and amplification of these genes. Our study reinforces the potential role of sinks in contributing to Enterobacterales infection and antimicrobial resistance in hospital patients, something that could be amenable to intervention

An Overview of the 2014 ALMA Long Baseline Campaign

A major goal of the Atacama Large Millimeter/submillimeter Array (ALMA) is to make accurate images with resolutions of tens of milliarcseconds, which at submillimeter (submm) wavelengths requires baselines up to ~15 km. To develop and test this capability, a Long Baseline Campaign (LBC) was carried out from September to late November 2014, culminating in end-to-end observations, calibrations, and imaging of selected Science Verification (SV) targets. This paper presents an overview of the campaign and its main results, including an investigation of the short-term coherence properties and systematic phase errors over the long baselines at the ALMA site, a summary of the SV targets and observations, and recommendations for science observing strategies at long baselines. Deep ALMA images of the quasar 3C138 at 97 and 241 GHz are also compared to VLA 43 GHz results, demonstrating an agreement at a level of a few percent. As a result of the extensive program of LBC testing, the highly successful SV imaging at long baselines achieved angular resolutions as fine as 19 mas at ~350 GHz. Observing with ALMA on baselines of up to 15 km is now possible, and opens up new parameter space for submm astronomy.Comment: 11 pages, 7 figures, 2 tables; accepted for publication in the Astrophysical Journal Letters; this version with small changes to affiliation

Capturing a Flavivirus Pre-Fusion Intermediate

During cell entry of flaviviruses, low endosomal pH triggers the rearrangement of the viral surface glycoproteins to a fusion-active state that allows the release of the infectious RNA into the cytoplasm. In this work, West Nile virus was complexed with Fab fragments of the neutralizing mAb E16 and was subsequently exposed to low pH, trapping the virions in a pre-fusion intermediate state. The structure of the complex was studied by cryo-electron microscopy and provides the first structural glimpse of a flavivirus fusion intermediate near physiological conditions. A radial expansion of the outer protein layer of the virion was observed compared to the structure at pH 8. The resulting ∼60 Å-wide shell of low density between lipid bilayer and outer protein layer is likely traversed by the stem region of the E glycoprotein. By using antibody fragments, we have captured a structural intermediate of a virus that likely occurs during cell entry. The trapping of structural transition states by antibody fragments will be applicable for other processes in the flavivirus life cycle and delineating other cellular events that involve conformational rearrangements

Caecal metastasis from breast cancer presenting as intestinal obstruction

<p>Abstract</p> <p>Background</p> <p>Gastrointestinal metastsasis from the breast cancer are rare. We report a patient who presented with intestinal obstruction due to solitary caecal metastasis from infiltrating ductal carcinoma of breast. We also review the available literature briefly.</p> <p>Case presentation</p> <p>A 72 year old lady with past history of breast cancer presented with intestinal obstruction due to a caecal mass. She underwent an emergency right hemicolectomy. The histological examination of the right hemicolectomy specimen revealed an adenocarcinoma in caecum staining positive for Cytokeratin 7 and Carcinoembryonic antigen and negative for Cytokeratin 20, CDX2 and Estrogen receptor. Eight out of 11 mesenteric nodes showed tumour deposits. A histological diagnosis of metastatic breast carcinoma was given.</p> <p>Conclusion</p> <p>To the best of our knowledge, this is the first case report of solitary metastasis to caecum from infiltrating ductal carcinoma of breast. Awareness of this possibility will aid in appropriate management of such patients.</p