Protein A-mediated binding of staphylococcus spp. to antibodies in flow cytometric assays and reduction of this binding by using Fc receptor blocking reagent

peer-reviewedStaphylococcus aureus and other coagulase-positive Staphylococcus spp. bind the Fc region of IgG antibodies through expression of protein A (SpA). These species have consequently been a source of false-positive signals in antibody-based assays designed to detect other target bacteria. Here, flow cytometry was used to study the influence of a number of factors on the SpA-mediated binding of single cells to an anti-human IgG antibody, including strain, heat killing, overnight storage, growth phase, cell physiology, surface adhesion, and growth in model food systems. Through the costaining of antibody-stained cells with the permeability dye pr opidium iodide and calcein violet AM, the cell physiological status was related to SpA-mediated antibody binding. Generally, permeabilized cells lacking esterase activity did not strongly bind antibody. The binding of a number of commercially available polyclonal IgG antibodies to non-Staphylococcus spp. was also characterized. Not all SpA-expressing species showed strong binding of mouse IgG, and one species not known to express SpA showed strong binding. Most SpA-expressing strains bound rabbit IgG antibodies to some extent, whereas only one strain bound goat IgG. To reduce or eliminate SpA-mediated IgG binding, the following products were evaluated as blocking reagents and applied prior to staining with primary or secondary antibody: normal rabbit serum, mouse IgG isotype control, goat IgG, and a commercial FcR blocking reagent. Only the FcR blocking reagent consistently reduced SpA-mediated binding of Staphylococcus spp. to antibodies against other species and could be recommended as a blocking reagent in immunoassays designed to detect non-Staphylococcus species

Redefining the effect of salt on thermophilic starter cell viability, culturability and metabolic activity in cheese

peer-reviewedThis study investigated the differential effect of salt concentration in the outside and inside layers of brine salted cheeses on viability, culturability and enzyme activity of starter bacteria. The high-salt environment of the outside layer caused a sharp decrease in L. helveticus viability as measured by traditional plate counts. Remarkably, this was associated with lower release of intracellular enzymes (LDH), reduced levels of proteolysis and larger membrane integrity as measured by flow cytometry (FC) following classical Live/Dead staining. FC analysis of light scattering properties highlighted a significant reduction in size and granularity of the microbiota located in the cheese surface, suggestive of cell shrinkage and condensation of internal macromolecules probably due to hyperosmotic stress. The microbiota of the cheese surface were found to experience greater oxidative stress, as measured by FC analysis of the total levels of reactive oxygen species, compared to that of the interior layer. These results lead us to postulate that the physiology and health status of the microbiota were significantly different in the outer and inner layers of the cheese. The hyperosmotic environment of the outer layer resulted in reduced cell lysis, as measurable by assays based upon membrane integrity, but rather triggered cell death via mechanisms involving cell shrinkage and ROS-mediated damage of vital intracellular components. This study challenges the current thinking on how salt controls microbial activity in ripening cheese, especially in cheeses which are brine salted as local variations in biochemical ripening indices can differ significantly from the outside to the inside of a ripening cheese

In search of Robert Bruce, part I: craniofacial analysis of the skull excavated at Dunfermline in 1819

Robert Bruce, king of Scots, is a significant figure in Scottish history, and his facial appearance will have been key to his status, power and resilience as a leader. This paper is the first in a series that discusses the burial and skeletal remains excavated at Dunfermline in 1819. Parts II and III discuss the evidence relating to whether or not the burial vault and skeleton belong to Robert Bruce, and Part I analyses and interprets the historical records and skeletal structure in order to produce a depiction of the facial appearance of Robert Bruce

Influence of addition of plasmin or mastitic milk to cheesemilk on quality of smear-ripened cheese

peer-reviewedSmear-ripened cheese varieties are characterised by the growth of a smear culture, containing predominantly Brevibacterium linens, on the cheese surface during ripening. In such cheese, considerable zonal differences in biochemistry of ripening exist, due to moisture loss from, and growth and metabolic activity of smear microflora at, the cheese surface. In this study, the effects of adding exogenous plasmin or small amounts of mastitic milk to good quality milk on the quality of smear-ripened cheese made subsequently was examined. Addition of plasmin did not influence cheese composition immediately after manufacture, but slightly decreased the rate of moisture loss during cheese ripening. Plasmin activity decreased during the early stages of ripening, but subsequently increased towards the end of ripening, perhaps due to changing pH conditions in the cheese. Addition of plasmin increased rates of primary proteolysis in cheese, as measured by levels of pH 4.6- soluble N and urea-PAGE, although production of later products of proteolysis appeared less affected. Addition ofmastitic milk had largely similar effects to addition of exogenous plasmin, which may reflect a high content of plasmin or plasminogen activators in such milk. Overall, changes in milk quality and enzymology appear to influence the quality of smear-ripened cheese

Influence of buttermilk powder or buttermilk addition on phospholipid content, chemical and bio-chemical composition and bacterial viability in Cheddar style-cheese

peer-reviewedThe effect of buttermilk powder addition post-curd formation or buttermilk addition to cheese milk on total and individual phospholipid content, chemical composition, enzyme activity, microbial populations and microstructure within Cheddar-style cheese was investigated. Buttermilk or buttermilk powder addition resulted in significant increases in total phospholipid content and their distribution throughout the cheese matrix. Addition of 10% buttermilk powder resulted in higher phospholipid content, moisture, pH and salt in moisture levels, and lower fat, fat in dry matter, L. helveticus and non-starter bacteria levels in cheeses. Buttermilk powder inclusion resulted in lower pH 4.6/Soluble Nitrogen (SN) levels and significantly lower free amino acid levels in 10% buttermilk powder cheeses. Buttermilk addition provided a more porous cheese microstructure with greater fat globule coalescence and increased free fat pools, while also increasing moisture and decreasing protein, fat and pH levels. Addition of buttermilk in liquid or powdered form offers potential for new cheeses with associated health benefits

Canonical thermalization

For quantum systems that are weakly coupled to a much 'bigger' environment, thermalization of possibly far from equilibrium initial ensembles is demonstrated: for sufficiently large times, the ensemble is for all practical purposes indistinguishable from a canonical density operator under conditions that are satisfied under many, if not all, experimentally realistic conditions

Effect of salt and fat reduction on proteolysis, rheology and cooking properties of Cheddar cheese

peer-reviewedFull-fat (FF, 33%), reduced-fat (RF, 22%) and half-fat (HF, 16%) Cheddar-style cheeses, each with full salt (FS, 1.9%), reduced salt (RS, 1.2%) or half salt (HS, 0.9%) variants, were made on three separate occasions. Reducing salt from 1.9 to 1.2 or 0.9% in the FF, RF and HF cheeses resulted in lower intact casein, firmness, fracture stress and work to extend the molten cheese, and higher heat-induced flow. The fracture strain of the RF and HF cheeses increased while that of the FF cheese decreased on reducing salt content. The results show the response to reducing salt depended on the variable studied, extent of salt reduction and fat level. Generally, reducing fat had opposite effects to those of salt reduction. There was a significant interaction between salt and fat content on the fracture characteristics, heat-induced flow, and extensibility of molten cheese at ripening times ≥90 d

A Keck/DEIMOS spectroscopic survey of the faint M31 satellites And IX, And XI, And XII, and And XIII

We present the first spectroscopic analysis of the faint M31 satellite galaxies, AndXI and AndXIII, and a reanalysis of existing spectroscopic data for two further faint companions, And IX and AndXII. By combining data obtained using the DEIMOS spectrograph mounted on the Keck II telescope with deep photometry from the Suprime-Cam instrument on Subaru, we have calculated global properties for the dwarfs, such as systemic velocities, metallicites and half-light radii.We find each dwarf to be very metal poor ([Fe/H] -2 both photometrically and spectroscopically, from their stacked spectrum), and as such, they continue to follow the luminosity-metallicity relationship established with brighter dwarfs. We are unable to resolve a dispersion for And XI due to small sample size and low S/N, but we set a one sigma upper limit of sigma-v <5 km/s. For And IX, And XII and And XIII we resolve velocity dispersions of v=4.5 (+3.4,-3.2), 2.6(+5.1,-2.6) and 9.7(+8.9,-4.5) km/s, and derive masses within the half light radii of 6.2(+5.3,-5.1)x10^6 Msun, 2.4 (+6.5,-2.4)x10^6 Msun and 1.1(+1.4,-0.7)x10^7 Msun respectively. We discuss each satellite in the context of the Mateo relations for dwarf spheroidal galaxies, and the Universal halo profiles established for Milky Way dwarfs (Walker et al. 2009). For both galaxies, this sees them fall below the Universal halo profiles of Walker et al. (2009). When combined with the findings of McConnachie & Irwin (2006a), which reveal that the M31 satellites are twice as extended (in terms of both half-light and tidal radii) as their Milky Way counterparts, these results suggest that the satellite population of the Andromeda system could inhabit halos that are significantly different from those of the Milky Way in terms of their central densities (abridged).Comment: 26 pages, 18 figures, MNRAS submitte

Depolarization and cAMP Elevation Rapidly Recruit TrkB to the Plasma Membrane of CNS Neurons

AbstractHere, we describe a novel mechanism for the rapid regulation of surface levels of the neurotrophin receptor TrkB. Unlike nodose ganglion neurons, both retinal ganglion cells (RGCs) and spinal motor neurons (SMNs) in culture display only low levels of surface TrkB, though high levels are present intracellularly. Within minutes of depolarization or cAMP elevation, surface TrkB levels increase by nearly 4-fold, and this increase is not blocked by cycloheximide. These findings suggest that activity and cAMP elevation rapidly recruit TrkB to the plasma membrane by translocation from intracellular stores. We propose that a fundamental difference between peripheral nervous system (PNS) and central nervous system (CNS) neurons is the activity dependence of CNS neurons for responsiveness to their peptide trophic factors and that differences in membrane compartmentalization of the receptors underlie this difference

First results from the Very Small Array -- IV. Cosmological parameter estimation

We investigate the constraints on basic cosmological parameters set by the first compact-configuration observations of the Very Small Array (VSA), and other cosmological data sets, in the standard inflationary LambdaCDM model. Using a weak prior 40 < H_0 < 90 km/s/Mpc and 0 < tau < 0.5 we find that the VSA and COBE_DMR data alone produce the constraints Omega_tot = 1.03^{+0.12}_{-0.12}, Omega_bh^2 = 0.029^{+0.009}_{-0.009}, Omega_cdm h^2 = 0.13^{+0.08}_{-0.05} and n_s = 1.04^{+0.11}_{-0.08} at the 68 per cent confidence level. Adding in the type Ia supernovae constraints, we additionally find Omega_m = 0.32^{+0.09}_{-0.06} and Omega_Lambda = 0.71^{+0.07}_{-0.07}. These constraints are consistent with those found by the BOOMERanG, DASI and MAXIMA experiments. We also find that, by combining all the recent CMB experiments and assuming the HST key project limits for H_0 (for which the X-ray plus Sunyaev--Zel'dovich route gives a similar result), we obtain the tight constraints Omega_m=0.28^{+0.14}_{-0.07} and Omega_Lambda= 0.72^{+0.07}_{-0.13}, which are consistent with, but independent of, those obtained using the supernovae data.Comment: 10 pages, 6 figures, MNRAS in pres