FORMATION OF SLOW-REACTING SUBSTANCE OF ANAPHYLAXIS IN HUMAN LUNG TISSUE AND CELLS BEFORE RELEASE

The capacity to extract slow-reacting substance of anaphylaxis (SRS-A) from human lung tissue or cells after immunologic activation, together with the measurement of SRS-A in both the extract and the surrounding fluid, permits study of total SRS-A generation. That the material extracted is SRS-A was established by both differential bioassay and purification. SRS-A accumulation was entirely intracellular after limited IgE-dependent direct or reversed anaphylactic activation. Intracellular accumulation also generally preceded release, with generation of SRS-A continuing well beyond a plateau in the cellular SRS-A level and the release of preformed mediators. The quantity of SRS-A generated after immunologic activation was modulated by the introduction of exogenous cyclic nucleotides, revealing a site of cyclic nucleotide action distinct from that on mediator release. The capacity to determine not only the release of preformed mediators but also the generation of a newly formed mediator, the sum of SRS-A in cells and supernate, adds an additional dimension to the analysis of the cellular events of immediate hypersensitivity

The Toll-Like receptor adaptor TRIF contributes to otitis media pathogenesis and recovery

<p>Abstract</p> <p>Background</p> <p>Toll-like receptor (TLR) signalling is crucial for innate immune responses to infection. The involvement of TLRs in otitis media (OM), the most prevalent childhood disease in developed countries, has been implicated by studies in middle ear cell lines, by association studies of TLR-related gene polymorphisms, and by altered OM in mice bearing mutations in TLR genes. Activated TLRs signal via two alternative intracellular signaling molecules with differing effects; MyD88 (Myeloid differentiation primary response gene 88) inducing primarily interleukin expression and TRIF (Tir-domain-containing adaptor inducing interferon β) mediating type I interferon (IFN) expression. We tested the hypothesis that TRIF and type I IFN signaling play a role in OM, using a murine model of OM induced by non-typeable <it>Haemophilus influenzae </it>(NTHi). The ME inflammatory response to NTHi was examined in wild-type (WT) and TRIF-/- mice by qPCR, gene microarray, histopathology and bacterial culture.</p> <p>Results</p> <p>Expression of TRIF mRNA was only modesty enhanced during OM, but both type I IFN signalling genes and type I IFN-inducible genes were significantly up-regulated in WT mice. TRIF-deficient mice showed reduced but more persistent mucosal hyperplasia and less leukocyte infiltration into the ME in response to NTHi infection than did WT animals. Viable bacteria could be cultured from MEs of TRIF-/- mice for much longer in the course of disease than was the case for middle ears of WT mice.</p> <p>Conclusion</p> <p>Our results demonstrate that activation of TRIF/type I IFN responses is important in both the pathogenesis and resolution of NTHi-induced OM.</p

The Evolution Of LMC X-4 Flares: Evidence For Super-Eddington Radiation Oozing Through Inhomogeneous Polar Cap Accretion Flows ?

We present the results of two extensive Rossi X-ray Timing Explorer observations of large X-ray flaring episodes from the high-mass X-ray binary pulsar LMC X-4. Light curves during the flaring episodes comprise bright peaks embedded in relatively fainter regions, with complex patterns of recurrence and clustering of flares. We identify precursors preceding the flaring activity. Pulse profiles during the flares appear to be simple sinusoids, and pulsed fractions are proportional to the flare intensities. We fit Gaussian functions to flare peaks to estimate the mean full-width-half-maximum to be $\sim$68 s. Significant rapid aperiodic variability exists up to a few hertz during the flares, which is related to the appearance of narrow, spiky peaks in the light curves. While spectral fits and softness ratios show overall spectral softening as the flare intensity increases, the narrow, spiky peaks do not follow this trend. The mean fluence of the flare peaks is (3.1 $\pm$ 2.9) $\times$ 10$^{40}$ ergs in the 2.5--25 keV energy range, with its maximum at $\sim$1.9 $\times$ 10$^{41}$ ergs. The flare peak luminosity reaches up to (2.1 $\pm$ 0.2) $\times$ 10$^{39}$ ergs s$^{-1}$, far above the Eddington luminosity of a neutron star. We discuss possible origins of the flares, and we also propose that inhomogeneous accretion columns onto the neutron star polar caps are responsible for the observed properties.Comment: 39 pages (including figures and tables), accepted for publication in Ap

SMC X-1 As An Intermediate-Stage Flaring X-ray Pulsar

We present Rossi X-ray Timing Explorer observations of the X-ray pulsar SMC X-1. The source is highly variable on short time scales (< 1 h), exhibiting apparent X-ray flares occupying a significant fraction (~3 %) of the total observing time, with a recurrence time of ~100 s. The flares seem to occur over all binary orbital phases, and correlate with the overall variability in the light curve. We find a total of 323 discrete flares which have a mean full width half maximum of ~18 s. The detailed properties of SMC X-1 do not vary significantly between the flares and the normal state, suggesting that the flare may be an extension of the normal state persistent emission with increased accretion rates. The flares resemble Type II X-ray bursts from GRO J1744--28. We discuss the origin of the SMC X-1 flares in terms of a viscous instability near the inner edge of the accretion disk around a weakly magnetized X-ray pulsar, and find this is consistent with the interpretation that SMC X-1 is in fact an intermediate-stage source like GRO J1744--28.Comment: 14 pages (5 figures), To appear in ApJ Letter

TNFA deletion alters apoptosis as well as caspase 3 and 4 expression during otitis media

Abstract Background Tumor necrosis factor (TNFA) is the canonical member of the TNF superfamily, which plays a major role in both inflammation and apoptosis. To evaluate the role of TNFs in otitis media (OM), the most common disease of childhood, we evaluated middle ear (ME) expression of genes encoding the TNF and TNF receptor superfamilies during bacterial OM in the mouse, characterized OM in TNFA-deficient mice, and assessed apoptosis during OM in normal versus TNF-deficient MEs. Results TNFs and TNF receptors were broadly regulated during OM, with TNFA showing the highest level of up-regulation. TNF deficient mice exhibited mucosal hyperplasia even in the absence of infection and exuberant growth of the mucosa during OM, including the formation of mucosal polyps. Mucosal recovery during OM was also delayed, in parallel with a delay in mucosal apoptosis and reduced caspase gene expression. Conclusions The TNF and TNF receptor superfamilies mediate both inflammation and apoptosis during OM. TNF appears to be critical for the maintenance of mucosal architecture in both the normal and infected ME, since excessive accumulation of mucosal tissue is seen in TNFA-/- MEs both before and after bacterial inoculation of the ME. TNFA is also required for appropriate regulation of caspase genes

The transcriptome of a complete episode of acute otitis media

Abstract Background Otitis media is the most common disease of childhood, and represents an important health challenge to the 10-15% of children who experience chronic/recurrent middle ear infections. The middle ear undergoes extensive modifications during otitis media, potentially involving changes in the expression of many genes. Expression profiling offers an opportunity to discover novel genes and pathways involved in this common childhood disease. The middle ears of 320 WBxB6 F1 hybrid mice were inoculated with non-typeable Haemophilus influenzae (NTHi) or PBS (sham control). Two independent samples were generated for each time point and condition, from initiation of infection to resolution. RNA was profiled on Affymetrix mouse 430 2.0 whole-genome microarrays. Results Approximately 8% of the sampled transcripts defined the signature of acute NTHi-induced otitis media across time. Hierarchical clustering of signal intensities revealed several temporal gene clusters. Network and pathway enrichment analysis of these clusters identified sets of genes involved in activation of the innate immune response, negative regulation of immune response, changes in epithelial and stromal cell markers, and the recruitment/function of neutrophils and macrophages. We also identified key transcriptional regulators related to events in otitis media, which likely determine the expression of these gene clusters. A list of otitis media susceptibility genes, derived from genome-wide association and candidate gene studies, was significantly enriched during the early induction phase and the middle re-modeling phase of otitis but not in the resolution phase. Our results further indicate that positive versus negative regulation of inflammatory processes occur with highly similar kinetics during otitis media, underscoring the importance of anti-inflammatory responses in controlling pathogenesis. Conclusions The results characterize the global gene response during otitis media and identify key signaling and transcription factor networks that control the defense of the middle ear against infection. These networks deserve further attention, as dysregulated immune defense and inflammatory responses may contribute to recurrent or chronic otitis in children

Coherence in the Quasi-Particle 'Scattering' by the Vortex Lattice in Pure Type-II Superconductors

The effect of quasi-particle (QP) 'scattering' by the vortex lattice on the de-Haas van-Alphen oscillations in a pure type-II superconductor is investigated within mean field,asymptotic perturbation theory. Using a 2D electron gas model it is shown that, due to a strict phase coherence in the many-particle correlation functions, the 'scattering' effect in the asymptotic limit ($\sqrt{E_F/\hbar\omega_c}\gg 1$) is much weaker than what is predicted by the random vortex lattice model proposed by Maki and Stephen, which destroys this coherence . The coherent many particle configuration is a collinear array of many particle coordinates, localized within a spatial region with size of the order of the magnetic length. The amplitude of the magnetization oscillations is sharply damped just below $$ because of strong $180^{\circ}$ out of phase magnetic oscillations in the superconducting condensation energy ,which tend to cancel the normal electron oscillations. Within the ideal 2D model used it is found, however, that because of the relative smallness of the quartic and higher order terms in the expansion, the oscillations amplitude at lower fields does not really damp to zero, but only reverses sign and remains virtually undamped well below $H_{c2}$. This conclusion may be changed if disorder in the vortex lattice, or vortex lines motion will be taken into account. The reduced QP 'scattering' effect may be responsible for the apparent crossover from a strong damping of the dHvA oscillations just below $H_{c2}$ to a weaker damping at lower fields observed experimentally in several 3D superconductors.Comment: 26 pages, Revtex no Figure

The role of DNA sensing and innate immune receptor TLR9 in otitis media

Otitis media (OM), a common infectious disease in children, is associated with bacterial middle ear (ME) infection. Tolllike receptors (TLRs) are important mediators of innate immune responses, and TLR9 specifically recognizes the unmethylated cytidine-phosphate-guanosine (CpG) motifs in bacterial DNA. Additional sensors of foreign DNA have recently been identified. The role of DNA sensing and TLR9 was investigated in a murine model of OM induced by non-typeable Haemophilus influenzae (NTHi). Expression of genes related to DNA-sensing pathways involved in innate immunity was assessed via DNA microarray, qPCR and immunohistochemistry. Middle ear responses to NTHi were examined in wild-type and TLR9−/− mice by histopathology and bacterial culture. Expression of TLR9 signaling genes was modestly up-regulated during OM, as was TLR9 protein in both ME mucosal cells and infiltrating leukocytes. However, genes known to be regulated by CpG DNA were dramatically up-regulated, as were genes involved in DNA sensing by DIA, Pol-III and AIM2. Toll-like receptor 9 deletion significantly prolonged the inflammatory response induced by NTHi in the ME and delayed bacterial clearance. The results suggest that DNA sensing via TLR9 plays a role in OM pathogenesis and recovery. Alternative forms of DNA sensing may also contribute to OM

TLR4-mediated induction of TLR2 signaling is critical in the pathogenesis and resolution of otitis media

Otitis media is the most prevalent childhood disease in developed countries. The involvement of Toll-like receptors (TLRs) in otitis media pathophysiology has been implicated by studies in cell lines and association studies of TLR gene polymorphisms. However, precise functions of TLRs in the etiology of otitis media in vivo have not been examined. We investigated the inflammatory response to nontypeable Haemophilus influenzae using a model of otitis media in wild-type, TLR2−/− and TLR4−/− mice by gene microarray, qPCR, immunohistochemistry, Western blot analysis and histopathology. Toll-like receptor-2−/− and TLR4−/− mice exhibited a more profound, persistent inflammation with impaired bacterial clearance compared to controls. While wild-type mice induced tumor necrosis factor-α (TNF) after non-typeable H. influenzae challenge, TLR2−/− and TLR4−/− mice lack TNF induction in the early phase of otitis media. Moreover, lack of TLR2 resulted in a late increase in IL-10 expression and prolonged failure to clear bacteria. Toll-like receptor-4−/− mice showed impaired early bacterial clearance and loss of TLR2 induction in early otitis media. Our results demonstrate that both TLR2 and TLR4 signalling are critical to the regulation of infection in non-typeable H. influenzae-induced otitis media. Toll-like receptor-4 signalling appears to induce TLR2 expression, and TLR2 activation is critical for bacterial clearance and timely resolution of otitis media