Characterization of an emergent clone of enteroinvasive Escherichia coli circulating in Europe

Enteroinvasive Escherichia coli (EIEC) cause intestinal illness indistinguishable from that caused by Shigella, mainly in developing countries. Recently an upsurge of cases of EIEC infections has been observed in Europe, with two large outbreaks occurring in Italy and in the United Kingdom. We have characterized phenotypically and genotypically the strains responsible for these epidemics together with an additional isolate from a sporadic case isolated in Spain. The three isolates belonged to the same rare serotype O96:H19 and were of sequence type ST-99, never reported before in EIEC or Shigella. The EIEC strains investigated possessed all the virulence genes harboured on the large plasmid conferring the invasive phenotype to EIEC and Shigella while showing only some of the known chromosomal virulence genes and none of the described pathoadaptative mutations. At the same time, they displayed motility abilities and biochemical requirements resembling more closely those of the non-pathogenic E. coli rather than the EIEC and Shigella strains used as reference. Our observations suggested that the O96:H19 strains belong to an emerging EIEC clone, which could be the result of a recent event of acquisition of the invasion plasmid by commensal E. coli

AAV-mediated and pharmacological induction of Hsp70 expression stimulates survival of retinal ganglion cells following axonal injury.

We evaluated the effect of AAV2- and 17-AAG (17-N-allylamino-17-demethoxygeldanamycin)-mediated upregulation of Hsp70 expression on the survival of retinal ganglion cells (RGCs) injured by optic nerve crush (ONC). AAV2-Hsp70 expression in the retina was primarily observed in the ganglion cell layer. Approximately 75% of all transfected cells were RGCs. RGC survival in AAV2-Hsp70-injected animals was increased by an average of 110% 2 weeks after the axonal injury compared with the control. The increase in cell numbers was not even across the retinas with a maximum effect of approximately 306% observed in the inferior quadrant. 17-AAG-mediated induction of Hsp70 expression has been associated with cell protection in various models of neurodegenerative diseases. We show here that a single intravitreal injection of 17-AAG (0.2 ug ul(-1)) results in an increased survival of ONC-injured RGCs by approximately 49% compared with the vehicle-treated animals. Expression of Hsp70 in retinas of 17-AAG-treated animals was upregulated approximately by twofold compared with control animals. Our data support the idea that the upregulation of Hsp70 has a beneficial effect on the survival of injured RGCs, and the induction of this protein could be viewed as a potential neuroprotective strategy for optic neuropathies

Cosmic-ray acceleration in supernova remnants: non-linear theory revised

A rapidly growing amount of evidences, mostly coming from the recent gamma-ray observations of Galactic supernova remnants (SNRs), is seriously challenging our understanding of how particles are accelerated at fast shocks. The cosmic-ray (CR) spectra required to account for the observed phenomenology are in fact as steep as $E^{-2.2}--E^{-2.4}$, i.e., steeper than the test-particle prediction of first-order Fermi acceleration, and significantly steeper than what expected in a more refined non-linear theory of diffusive shock acceleration. By accounting for the dynamical back-reaction of the non-thermal particles, such a theory in fact predicts that the more efficient the particle acceleration, the flatter the CR spectrum. In this work we put forward a self-consistent scenario in which the account for the magnetic field amplification induced by CR streaming produces the conditions for reversing such a trend, allowing --- at the same time --- for rather steep spectra and CR acceleration efficiencies (about 20%) consistent with the hypothesis that SNRs are the sources of Galactic CRs. In particular, we quantitatively work out the details of instantaneous and cumulative CR spectra during the evolution of a typical SNR, also stressing the implications of the observed levels of magnetization on both the expected maximum energy and the predicted CR acceleration efficiency. The latter naturally turns out to saturate around 10-30%, almost independently of the fraction of particles injected into the acceleration process as long as this fraction is larger than about $10^{-4}$.Comment: 24 pages, 5 figures, accepted for publication in JCA

Diffusive propagation of cosmic rays from supernova remnants in the Galaxy. I: spectrum and chemical composition

In this paper we investigate the effect of stochasticity in the spatial and temporal distribution of supernova remnants on the spectrum and chemical composition of cosmic rays observed at Earth. The calculations are carried out for different choices of the diffusion coefficient D(E) experienced by cosmic rays during propagation in the Galaxy. In particular, at high energies we assume that D(E)\sim E^{\delta}, with $\delta=1/3$ and $\delta=0.6$ being the reference scenarios. The large scale distribution of supernova remnants in the Galaxy is modeled following the distribution of pulsars, with and without accounting for the spiral structure of the Galaxy. We find that the stochastic fluctuations induced by the spatial and temporal distribution of supernovae, together with the effect of spallation of nuclei, lead to mild but sensible violations of the simple, leaky-box-inspired rule that the spectrum observed at Earth is $N(E)\propto E^{-\alpha}$ with $\alpha=\gamma+\delta$, where $\gamma$ is the slope of the cosmic ray injection spectrum at the sources. Spallation of nuclei, even with the small rates appropriate for He, may account for slight differences in spectral slopes between different nuclei, providing a possible explanation for the recent CREAM observations. For $\delta=1/3$ we find that the slope of the proton and helium spectra are $\sim 2.67$ and $\sim 2.6$ respectively at energies above 1 TeV (to be compared with the measured values of $2.66\pm 0.02$ and $2.58\pm 0.02$). For $\delta=0.6$ the hardening of the He spectra is not observed. We also comment on the effect of time dependence of the escape of cosmic rays from supernova remnants, and of a possible clustering of the sources in superbubbles. In a second paper we will discuss the implications of these different scenarios for the anisotropy of cosmic rays.Comment: 28 pages, To appear in JCA

Chemical Composition and Cytoprotective Activities of Methanolic Extract of Asplenium adiantum-nigrum L. (Aspleniaceae)

Ferns can be a source of polyphenolic compounds, with the fronds being the main parts described in ethnopharmacological studies. The present study screened polyphenolic phytochemicals and evaluated in vitro activities of a methanolic extract of Asplenium adiantum-nigrum L. fronds (AAM), an Aspleniaceae fern collected from the Prades mountains (Tarragona, Spain). Phytocharacterization by HPLC-MS/MS confirmed that the major flavonoids isolated in AAM are flavanols while the major phytochemicals are phenol acids, with chlorogenic acid being the most representative one. Cytotoxicity, cytoprotection, cellular repair activity, and phototoxicity were determined in vitro in the presence of 0.01, 0.1, and 1 mg/mL of the extract. No cytotoxicity was obtained in any of the cell lines tested: non-tumoral (3T3 and HaCaT) and tumoral (HeLa, HepG2, and A549) cells. Additionally, the polyphenolic extract showed greater protective effect against H2O2 in 3T3 than HaCaT cells. Despite the low total phenolic content of AAM (1405.68 mg phenolic phytochemicals/kg dry extract), the cytoprotective activity of this extract could be associated with the synergistic antioxidant action of their polyphenolic profile. In addition, the extract did not present phototoxicity against the non-cytotoxic 1.8 J/cm2 dose of UVA light in both non-tumoral cell lines

A new pathogenicity island carrying an allelic variant of the Subtilase cytotoxin is common among Shiga toxin producing Escherichia coli of human and ovine origin

AbstractSubtilase (SubAB) is a cytotoxin elaborated by some Shiga Toxin (Stx)-producing Escherichia coli (STEC) strains usually lacking the locus of enterocyte effacement (LEE). Two variants of SubAB coding genes have been described: subAB1, located on the plasmid of the STEC O113 98NK2 strain, and subAB2, located on a pathogenicity island (PAI) together with the tia gene, encoding an invasion determinant described in enterotoxigenic E. coli. In the present study, we determined the entire nucleotide sequence of the PAI containing the subAB2 operon, termed Subtilase-Encoding PAI (SE-PAI), and identified its integration site in the pheV tRNA locus. In addition, a PCR strategy for discriminating the two subAB allelic variants was developed and used to investigate their presence in E. coli strains belonging to different pathotypes and in a large collection of LEE-negative STEC of human and ovine origin. The results confirmed that subAB genes are carried predominantly by STEC and showed their presence in 72% and 86% of the LEE-negative strains from human cases of diarrhoea and from healthy sheep respectively. Most of the subAB-positive strains (98%) identified possessed the subAB2 allelic variant and were also positive for tia, suggesting the presence of SE-PAI. Altogether, our observations indicate that subAB2 is the prevalent SubAB-coding operon in LEE-negative STEC circulating in European countries, and that sheep may represent an important reservoir for human infections with these strains. Further studies are needed to assess the role of tia and/or other genes carried by SE-PAI in the colonization of the host intestinal mucosa

Diffusive propagation of cosmic rays from supernova remnants in the Galaxy. II: anisotropy

We investigate the effects of stochasticity in the spatial and temporal distribution of supernova remnants on the anisotropy of cosmic rays observed at Earth. The calculations are carried out for different choices of the diffusion coefficient D(E) for propagation in the Galaxy. The propagation and spallation of nuclei are taken into account. At high energies we assume that $D(E)\sim(E/Z)^{\delta}$, with $\delta=1/3$ and $\delta=0.6$ being the reference scenarios. The large scale distribution of supernova remnants in the Galaxy is modeled following the distribution of pulsars with and without accounting for the spiral structure of the Galaxy. Our calculations allow us to determine the contribution to anisotropy resulting from both the large scale distribution of SNRs in the Galaxy and the random distribution of the nearest remnants. The naive expectation that the anisotropy amplitude scales as D(E) is shown to be an oversimplification which does not reflect in the predicted anisotropy for any realistic distribution of the sources. The fluctuations in the anisotropy pattern are dominated by nearby sources, so that predicting or explaining the observed anisotropy amplitude and phase becomes close to impossible. We find however that the very weak energy dependence of the anisotropy amplitude below $10^{5}$ GeV and the rise at higher energies, can best be explained if the diffusion coefficient is $D(E)\sim E^{1/3}$. Faster diffusion, for instance with $\delta=0.6$, leads in general to an exceedingly large anisotropy amplitude. The spiral structure introduces interesting trends in the energy dependence of the anisotropy pattern, which qualitatively reflect the trend seen in the data. For large values of the halo size we find that the anisotropy becomes dominated by the large scale regular structure of the source distribution, leading indeed to a monotonic increase of $\delta_A$ with energy.Comment: 21 Pages, to appear in JCA