92 research outputs found
Vaccination against Clostridium difficile using toxin fragments: Observations and analysis in animal models
Clostridium difficile is a major cause of antibiotic associated diarrhea. Recently, we have shown that effective protection can be mediated in hamsters through the inclusion of specific recombinant fragments from toxin A and B in a systemically delivered vaccine. Interestingly while neutralizing antibodies to the binding domains of both toxin A and B are moderately protective, enhanced survival is observed when fragments from the glucosyltransferase region of toxin B replace those from the binding domain of this toxin. In this addendum, we discuss additional information that has been derived from such vaccination studies. This includes observations on efficacy and cross-protection against different ribotypes mediated by these vaccines and the challenges that remain for a vaccine which prevents clinical symptoms but not colonization. The use and value of vaccination both in the prevention of infection and for treatment of disease relapse will be discussed
Identification of oligomerization and drug-binding domains of the membrane fusion protein EmrA
Many pathogenic Gram-negative bacteria possess tripartite transporters that catalyze drug extrusion across the inner and outer membranes, thereby conferring resistance. These transporters consist of inner (IMP) and outer (OMP) membrane proteins, which are coupled by a periplasmic membrane fusion (MFP) protein. However, it is not know whether the MFP translocates the drug between the membranes, by acting as a channel, or whether it brings the IMP and OMP together, facilitating drug transfer. The MFP EmrA has an elongated periplasmic domain, which binds transported drugs, and is anchored to the inner membrane by a single -helix, which contains a leucine zipper dimerization domain. Consistent with CD and hydrodynamic analyses, the periplasmic domain is predicted to be composed of a -sheet subdomain and an -helical coiled-coil. We propose that EmrA forms a trimer in which the coiled-coils radiate across the periplasm, where they could sequester the OMP TolC. The "free" leucine zipper in the EmrA trimer might stabilize the interaction with the IMP EmrB, which also possesses leucine zipper motifs in the putative N- and C-terminal helices. The -sheet subdomain of EmrA would sit at the membrane surface adjacent to the EmrB, from which it receives the transported drug, inducing a conformational change that triggers the interaction with the OMP
Time-Dependent Multi-Centre Solutions from New Metrics with Holonomy Sim(n-2)
The classifications of holonomy groups in Lorentzian and in Euclidean
signature are quite different. A group of interest in Lorentzian signature in n
dimensions is the maximal proper subgroup of the Lorentz group, SIM(n-2).
Ricci-flat metrics with SIM(2) holonomy were constructed by Kerr and Goldberg,
and a single four-dimensional example with a non-zero cosmological constant was
exhibited by Ghanam and Thompson. Here we reduce the problem of finding the
general -dimensional Einstein metric of SIM(n-2) holonomy, with and without
a cosmological constant, to solving a set linear generalised Laplace and
Poisson equations on an (n-2)-dimensional Einstein base manifold. Explicit
examples may be constructed in terms of generalised harmonic functions. A
dimensional reduction of these multi-centre solutions gives new time-dependent
Kaluza-Klein black holes and monopoles, including time-dependent black holes in
a cosmological background whose spatial sections have non-vanishing curvature.Comment: Typos corrected; 29 page
Infection of hamsters with the UK Clostridium difficile ribotype 027 outbreak strain R20291
Clostridium difficile is the main cause of antibiotic-associated disease, a disease of high socio-economical importance that has recently been compounded by the global spread of the 027 (BI/NAP1/027) ribotype. C. difficile cases attributed to ribotype 027 strains have high recurrence rates (up to 36 %) and increased disease severity. The hamster model of infection is widely accepted as an appropriate model for studying aspects of C. difficile host–pathogen interactions. Using this model we characterized the infection kinetics of the UK 2006 outbreak strain, R20291. Hamsters were orally given a dose of clindamycin, followed 5 days later with 10 000 C. difficile spores. All 100 % of the hamsters succumbed to infection with a mean time to the clinical end point of 46.7 h. Colonization of the caecum and colon were observed 12 h post-infection reaching a maximum of approximately 3×104 c.f.u. per organ, but spores were not detected until 24 h post-infection. At 36 h post-infection C. difficile numbers increased significantly to approximately 6×107 c.f.u. per organ where numbers remained high until the clinical end point. Increasing levels of in vivo toxin production coincided with increases in C. difficile numbers in organs reaching a maximum at 36 h post-infection in the caecum. Epithelial destruction and polymorphonuclear leukocyte (PMN) recruitment occurred early on during infection (24 h) accumulating as gross microvilli damage, luminal PMN influx, and blood associated with mucosal muscle and microvilli. These data describe the fatal infection kinetics of the clinical UK epidemic C. difficile strain R20291 in the hamster infection model
Bile salt metabolism is not the only factor contributing to Clostridioides (Clostridium) difficile disease severity in the murine model of disease
Susceptibility of patients to antibiotic-associated C. difficile disease is intimately associated with specific changes to gut microbiome composition. In particular, loss of microbes that modify bile salt acids (BSA) play a central role; primary bile acids stimulate spore germination whilst secondary bile acids limit C. difficile vegetative growth. To determine the relative contribution of bile salt (BS) metabolism on C. difficile disease severity, we treated mice with three combinations of antibiotics prior to infection. Mice given clindamycin alone became colonized but displayed no tissue pathology while severe disease, exemplified by weight loss and inflammatory tissue damage occurred in animals given a combination of five antibiotics and clindamycin. Animals given only the five antibiotic cocktails showed only transient colonization and no disease. C. difficile colonization was associated with a reduction in bacterial diversity, an inability to amplify bile salt hydrolase (BSH) sequences from fecal DNA and a relative increase in primary bile acids (pBA) in cecal lavages from infected mice. Further, the link between BSA modification and the microbiome was confirmed by the isolation of strains of Lactobacillus murinus that modified primary bile acids in vitro, thus preventing C. difficile germination. Interestingly, BSH activity did not correlate with disease severity which appeared linked to alternations in mucin, which may indirectly lead to increased exposure of the epithelial surface to inflammatory signals. These data confirm the role of microbial metabolic activity in protection of the gut and highlights the need for greater understanding the function of bacterial communities in disease prevention
Susceptibility of hamsters to clostridium difficile isolates of differing toxinotype
Clostridium difficile is the most commonly associated cause of antibiotic associated disease (AAD), which caused ~21,000 cases of AAD in 2011 in the U.K. alone. The golden Syrian hamster model of CDI is an acute model displaying many of the clinical features of C. difficile disease. Using this model we characterised three clinical strains of C. difficile, all differing in toxinotype; CD1342 (PaLoc negative), M68 (toxinotype VIII) and BI-7 (toxinotype III). The naturally occurring non-toxic strain colonised all hamsters within 1-day post challenge (d.p.c.) with high-levels of spores being shed in the faeces of animals that appeared well throughout the entire experiment. However, some changes including increased neutrophil influx and unclotted red blood cells were observed at early time points despite the fact that the known C. difficile toxins (TcdA, TcdB and CDT) are absent from the genome. In contrast, hamsters challenged with strain M68 resulted in a 45% mortality rate, with those that survived challenge remaining highly colonised. It is currently unclear why some hamsters survive infection, as bacterial and toxin levels and histology scores were similar to those culled at a similar time-point. Hamsters challenged with strain BI-7 resulted in a rapid fatal infection in 100% of the hamsters approximately 26 hr post challenge. Severe caecal pathology, including transmural neutrophil infiltrates and extensive submucosal damage correlated with high levels of toxin measured in gut filtrates ex vivo. These data describes the infection kinetics and disease outcomes of 3 clinical C. difficile isolates differing in toxin carriage and provides additional insights to the role of each toxin in disease progression
On the `Stationary Implies Axisymmetric' Theorem for Extremal Black Holes in Higher Dimensions
All known stationary black hole solutions in higher dimensions possess
additional rotational symmetries in addition to the stationary Killing field.
Also, for all known stationary solutions, the event horizon is a Killing
horizon, and the surface gravity is constant. In the case of non-degenerate
horizons (non-extremal black holes), a general theorem was previously
established [gr-qc/0605106] proving that these statements are in fact generally
true under the assumption that the spacetime is analytic, and that the metric
satisfies Einstein's equation. Here, we extend the analysis to the case of
degenerate (extremal) black holes. It is shown that the theorem still holds
true if the vector of angular velocities of the horizon satisfies a certain
"diophantine condition," which holds except for a set of measure zero.Comment: 30pp, Latex, no figure
Medin aggregation causes cerebrovascular dysfunction in aging wild-type mice
Medin is the most common amyloid known in humans, as it can be found in blood vessels of the upper body in virtually everybody over 50 years of age. However, it remains unknown whether deposition of Medin plays a causal role in age-related vascular dysfunction. We now report that aggregates of Medin also develop in the aorta and brain vasculature of wild-type mice in an age-dependent manner. Strikingly, genetic deficiency of the Medin precursor protein, MFG-E8, eliminates not only vascular aggregates but also prevents age-associated decline of cerebrovascular function in mice. Given the prevalence of Medin aggregates in the general population and its role in vascular dysfunction with aging, targeting Medin may become a novel approach to sustain healthy aging
Bitter taste cells in the ventricular walls of the murine brain regulate glucose homeostasis
The median eminence (ME) is a circumventricular organ at the base of the
brain that controls body homeostasis. Tanycytes are its specialized glial cells
that constitute the ventricular walls and regulate different physiological states,
however individual signaling pathways in these cells are incompletely understood. Here, we identify a functional tanycyte subpopulation that expresses
key taste transduction genes including bitter taste receptors, the G protein
gustducin and the gustatory ion channel TRPM5 (M5). M5 tanycytes have
access to blood-borne cues via processes extended towards diaphragmed
endothelial fenestrations in the ME and mediate bidirectional communication
between the cerebrospinal fluid and blood. This subpopulation responds to
metabolic signals including leptin and other hormonal cues and is transcriptionally reprogrammed upon fasting. Acute M5 tanycyte activation
induces insulin secretion and acute diphtheria toxin-mediated M5 tanycyte
depletion results in impaired glucose tolerance in diet-induced obese mice. We
provide a cellular and molecular framework that defines how bitter taste cells
in the ME integrate chemosensation with metabolism
Synthesis and magnetic properties of NiFe_{2-x}Al_{x}O_{4} nanoparticles
Nanocrystalline Al-doped nickel ferrite powders have been synthesized by
sol-gel auto-ignition method and the effect of non-magnetic aluminum content on
the structural and magnetic properties has been studied. The X-ray diffraction
(XRD) revealed that the powders obtained are single phase with inverse spinel
structure. The calculated grain sizes from XRD data have been verified using
transmission electron microscopy (TEM). TEM photographs show that the powders
consist of nanometer-sized grains. It was observed that the characteristic
grain size decreases from 29 to 6 nm as the non-magnetic Al content increases,
which was attributed to the influence of non-magnetic Al concentration on the
grain size. Magnetic hysteresis loops were measured at room temperature with a
maximum applied magnetic field of 1T. As aluminum content increases, the
measured magnetic hysteresis curves become more and more narrow and the
saturation magnetization and remanent magnetization both decreased. The
reduction of agnetization compared to bulk is a consequence of spin
non-collinearity. Further reduction of magnetization with increase of aluminum
content is caused by non-magnetic Al^{3+} ions and weakened interaction between
sublattices. This, as well as the decrease in hysteresis was understood in
terms of the decrease in particle size.Comment: 24 pages, 6 figure
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