Nuclear-localized focal adhesion kinase regulates inflammatory VCAM-1 expression.

Vascular cell adhesion molecule-1 (VCAM-1) plays important roles in development and inflammation. Tumor necrosis factor-α (TNF-α) and focal adhesion kinase (FAK) are key regulators of inflammatory and integrin-matrix signaling, respectively. Integrin costimulatory signals modulate inflammatory gene expression, but the important control points between these pathways remain unresolved. We report that pharmacological FAK inhibition prevented TNF-α-induced VCAM-1 expression within heart vessel-associated endothelial cells in vivo, and genetic or pharmacological FAK inhibition blocked VCAM-1 expression during development. FAK signaling facilitated TNF-α-induced, mitogen-activated protein kinase activation, and, surprisingly, FAK inhibition resulted in the loss of the GATA4 transcription factor required for TNF-α-induced VCAM-1 production. FAK inhibition also triggered FAK nuclear localization. In the nucleus, the FAK-FERM (band 4.1, ezrin, radixin, moesin homology) domain bound directly to GATA4 and enhanced its CHIP (C terminus of Hsp70-interacting protein) E3 ligase-dependent polyubiquitination and degradation. These studies reveal new developmental and anti-inflammatory roles for kinase-inhibited FAK in limiting VCAM-1 production via nuclear localization and promotion of GATA4 turnover

Water content and morphology of sodium chloride aerosol particles

This is the publisher's version, also available electronically from http://onlinelibrary.wiley.com/doi/10.1029/1999JD900286/abstract;jsessionid=41C36E183A6316F5D3C491131615BD7A.f01t04.Sodium chloride droplets with a median diameter of ∼0.4 μm were generated in the laboratory by atomizing an aqueous solution of NaCl under ambient conditions. Infrared extinction spectra of the aerosols under controlled relative humidity (RH) ranging from 15 to 95% were obtained. The extinction spectra contained both scattering and absorption components. In order to obtain an absorption spectrum of the condensed phase H2O associated with the particulates, it was necessary to subtract from the extinction spectra the absorption by H2O vapor and the scattering by the particulates. H2O vapor subtraction was accomplished by a standard technique. A procedure using Mie theory to subtract the scattering component of the extinction spectrum is described. The absorption spectra were used to determine the water content and structure of the particulates. Above ∼50% RH the aerosols contain aqueous droplets that have not reached equilibrium with the water vapor during the timescale of the experiments (∼10 s). There is a sharp transition in water content at around 50% RH which is consistent with other measures of the recrystallization point. Below 50% RH the NaCl particles contain an anomalously large amount of H2O. Several different particle models are considered to explain the H2O content. The model in which the NaCl particles contain pockets of aqueous NaCl solution was found to be most consistent with the spectroscopic observations. The relevance of salt particle morphology and water content to atmospheric aerosol chemistry is discussed

Infrared spectroscopic signatures of (NH4)2SO4 aerosols

This is the publisher's version, also available electronically from http://onlinelibrary.wiley.com/doi/10.1029/96JD01543/abstract.Ammonium sulfate particles in air with average diameters ranging from 0.1 to 0.5-μm have been generated by atomizing aqueous solutions of (NH4)2SO4 of various concentrations at ambient temperatures and pressures. The infrared spectra from 4000 to 600 cm−1 of the resulting aerosols have been investigated. This spectral region has allowed us to study the four infrared-active vibrational modes of this salt: ν3(NH4+), ν4(NH4+), ν3(SO42−), and ν4(SO42−). The frequencies of these modes are similar to published results obtained from infrared studies of the single crystal but are displaced to higher wavenumbers. Depending on relative humidity, the aerosol particles are crystalline or supersaturated aqueous droplets. These phase identifications are possible because liquid water absorption features are found in the droplets but not in the crystals. Extensive Mie theory calculations have been performed for spheres of diameters ranging from 0.1-μm to 2.0-μm to explore frequency shifts and the relative contributions to extinction of scattering and absorption with particle size. We show that, for the smaller particles, the molecular cross section in the ν3(SO42−) region can be used to determine the number of (NH4)2SO4 molecules in an aerosol sample. The (small) frequency shifts in this region provide information on the aerosol particle size. A Mie theory calculation of extinction for a model polydisperse aerosol, believed to approximate that of an experimental aerosol, gives reasonable agreement with the observed spectrum. While calculated band centers of the four modes are within 1% of those observed, values of extinction can differ by as much as 50%. We discuss possible reasons for the discrepancies. Spectroscopic changes observed for an aerosol as the particles settle are discussed in terms of kinetic models and Mie theory. We discuss the potential of spectroscopic signatures of tropospheric (NH4)2SO4 aerosols for the characterization of their size, morphology, phase, and composition. Finally, we propose a field experiment to measure sulfate aerosol in the arctic troposphere

Label-free, in-solution screening of peptide libraries for binding to protein targets using hydrogen exchange-mass spectrometry (HX-MS)

There is considerable interest in the discovery of peptide ligands that bind to protein targets. Discovery of such ligands is usually approached by screening large peptide libraries. However, the individual peptides must be tethered to a tag that preserves their individual identities (e.g. phage display or one-bead one-compound). To overcome this limitation, we have developed a method for screening libraries of label-free peptides for binding to a protein target in solution as a single batch. The screening is based on decreased amide hydrogen exchange by peptides that bind to the target. Hydrogen exchange was measured by mass spectrometry. We demonstrate the approach using a peptide library derived from the E. coli proteome that contained 6664 identifiable features. The library was spiked separately with a peptide spanning the calmodulin binding domain of endothelial nitric oxide synthase (eNOS, 494-513) and a peptide spanning the N-terminal twenty residues of bovine ribonuclease A (S peptide). Human calmodulin and bovine ribonuclease S (RNase S) were screened against the library. Using a novel data analysis workflow we identified the eNOS peptide as the only calmodulin binding peptide and S peptide as the only ribonuclease S binding peptide in the library

Sr-Nd-Pb-Hf isotope results from ODP Leg 187: Evidence for mantle dynamics of the Australian-Antarctic Discordance and origin of the Indian MORB source

New high precision PIMMS Hf and Pb isotope data for 14–28 Ma basalts recovered during ODP Leg 187 are compared with zero-age dredge samples from the Australian-Antarctic Discordance (AAD). These new data show that combined Nd-Hf isotope systematics can be used as an effective discriminant between Indian and Pacific MORB source mantle domains. In particular, Indian mantle is displaced to lower εNd and higher εHf ratios compared to Pacific mantle. As with Pb isotope plots, there is almost no overlap between the two mantle types in Nd-Hf isotope space. On the basis of our new Nd-Hf isotope data, we demonstrate that Pacific MORB-source mantle was present near the eastern margin of the AAD from as early as 28 Ma, its boundary with Indian MORB-source mantle coinciding with the eastern edge of a basin-wide arcuate depth anomaly that is centered on the AAD. This observation rules out models requiring rapid migration of Pacific MORB mantle into the Indian Ocean basin since separation of Australia from Antarctica. Although temporal variations in isotopic composition can be discerned relative to the fracture zone boundary of the modern AAD at 127°E, the distribution of different compositional groups appears to have remained much the same relative to the position of the residual depth anomaly for the past 30 m.y. Thus significant lateral flow of mantle along the ridge axis toward the interface appears unlikely. Instead, the dynamics that maintain both the residual depth anomaly and the isotopic boundary between Indian and Pacific mantle are due to eastward migration of the Australian and Antarctic plates over a stagnated, but slowly upwelling, slab oriented roughly orthogonal to the ridge axis. Temporal and spatial variations in the compositions of Indian MORB basalts within the AAD can be explained by progressive displacement of shallower Indian MORB-source mantle by deeper mantle having a higher εHf composition ascending ahead of the upwelling slab. Models for the origin of the distinctive composition of the Indian MORB-source based on recycling of a heterogeneous enriched component that consist of ancient altered ocean crust plus<10% pelagic sediment are inconsistent with Nd-Hf isotope systematics. Instead, the data can be explained by a model in which Indian mantle includes a significant proportion of material that was processed in the mantle wedge above a subduction zone and was subsequently mixed back into unprocessed upper mantle

An initial event in insect innate immune response: structural and biological studies of interactions between β-1,3-glucan and the N-terminal domain of β-1,3-glucan recognition protein

In response to invading microorganisms, insect β-1,3-glucan recognition protein (βGRP), a soluble receptor in the hemolymph, binds to the surfaces of bacteria and fungi and activates serine protease cascades that promote destruction of pathogens by means of melanization or expression of antimicrobial peptides. Here we report on the NMR solution structure of the N-terminal domain of βGRP (N-βGRP) from Indian meal moth (Plodia interpunctella), which is sufficient to activate the prophenoloxidase (proPO) pathway resulting in melanin formation. NMR and isothermal calorimetric titrations of N-βGRP with laminarihexaose, a glucose hexamer containing β-1,3 links, suggest a weak binding of the ligand. However, addition of laminarin, a glucose polysaccharide (~ 6 kDa) containing β-1,3 and β-1,6 links that activates the proPO pathway, to N-βGRP results in the loss of NMR cross-peaks from the backbone 15N-1H groups of the protein, suggesting the formation of a large complex. Analytical ultra centrifugation (AUC) studies of formation of N-βGRP:laminarin complex show that ligand-binding induces sel-fassociation of the protein:carbohydrate complex into a macro structure, likely containing six protein and three laminarin molecules (~ 102 kDa). The macro complex is quite stable, as it does not undergo dissociation upon dilution to sub-micromolar concentrations. The structural model thus derived from the present studies for N-βGRP:laminarin complex in solution differs from the one in which a single N-βGRP molecule has been proposed to bind to a triple helical form of laminarin on the basis of an X-ray crystallographic structure of N-βGRP:laminarihexaose complex [Kanagawa, M., Satoh, T., Ikeda, A., Adachi, Y., Ohno, N., and Yamaguchi, Y. (2011) J. Biol. Chem. 286, 29158-29165]. AUC studies and phenoloxidase activation measurements carried out with the designed mutants of N-βGRP indicate that electrostatic interactions involving Asp45, Arg54, and Asp68 between the ligand-bound protein molecules contribute in part to the stability of N-βGRP:laminarin macro complex and that a decreased stability is accompanied by a reduced activation of the proPO pathway. Increased β-1,6 branching in laminarin also results in destabilization of the macro complex. These novel findings suggest that ligand-induced self-association of βGRP:β-1,3-glucan complex may form a platform on a microbial surface for recruitment of downstream proteases, as a means of amplification of the initial signal of pathogen recognition for the activation of the proPO pathway

Climate change promotes parasitism in a coral symbiosis.

Coastal oceans are increasingly eutrophic, warm and acidic through the addition of anthropogenic nitrogen and carbon, respectively. Among the most sensitive taxa to these changes are scleractinian corals, which engineer the most biodiverse ecosystems on Earth. Corals' sensitivity is a consequence of their evolutionary investment in symbiosis with the dinoflagellate alga, Symbiodinium. Together, the coral holobiont has dominated oligotrophic tropical marine habitats. However, warming destabilizes this association and reduces coral fitness. It has been theorized that, when reefs become warm and eutrophic, mutualistic Symbiodinium sequester more resources for their own growth, thus parasitizing their hosts of nutrition. Here, we tested the hypothesis that sub-bleaching temperature and excess nitrogen promotes symbiont parasitism by measuring respiration (costs) and the assimilation and translocation of both carbon (energy) and nitrogen (growth; both benefits) within Orbicella faveolata hosting one of two Symbiodinium phylotypes using a dual stable isotope tracer incubation at ambient (26 °C) and sub-bleaching (31 °C) temperatures under elevated nitrate. Warming to 31 °C reduced holobiont net primary productivity (NPP) by 60% due to increased respiration which decreased host %carbon by 15% with no apparent cost to the symbiont. Concurrently, Symbiodinium carbon and nitrogen assimilation increased by 14 and 32%, respectively while increasing their mitotic index by 15%, whereas hosts did not gain a proportional increase in translocated photosynthates. We conclude that the disparity in benefits and costs to both partners is evidence of symbiont parasitism in the coral symbiosis and has major implications for the resilience of coral reefs under threat of global change

Measurement-based quantum control of mechanical motion

Controlling a quantum system based on the observation of its dynamics is inevitably complicated by the backaction of the measurement process. Efficient measurements, however, maximize the amount of information gained per disturbance incurred. Real-time feedback then enables both canceling the measurement's backaction and controlling the evolution of the quantum state. While such measurement-based quantum control has been demonstrated in the clean settings of cavity and circuit quantum electrodynamics, its application to motional degrees of freedom has remained elusive. Here we show measurement-based quantum control of the motion of a millimetre-sized membrane resonator. An optomechanical transducer resolves the zero-point motion of the soft-clamped resonator in a fraction of its millisecond coherence time, with an overall measurement efficiency close to unity. We use this position record to feedback-cool a resonator mode to its quantum ground state (residual thermal occupation n = 0.29 +- 0.03), 9 dB below the quantum backaction limit of sideband cooling, and six orders of magnitude below the equilibrium occupation of its thermal environment. This realizes a long-standing goal in the field, and adds position and momentum to the degrees of freedom amenable to measurement-based quantum control, with potential applications in quantum information processing and gravitational wave detectors.Comment: New version with corrected detection efficiency as determined with a NIST-calibrated photodiode, added references and revised structure. Main conclusions are identical. 41 pages, 18 figure

Sub-Doppler spectroscopy of Rb atoms in a sub-micron vapor cell in the presence of a magnetic field

We report the first use of an extremely thin vapor cell (thickness ~ 400 nm) to study the magnetic-field dependence of laser-induced-fluorescence excitation spectra of alkali atoms. This thin cell allows for sub-Doppler resolution without the complexity of atomic beam or laser cooling techniques. This technique is used to study the laser-induced-fluorescence excitation spectra of Rb in a 50 G magnetic field. At this field strength the electronic angular momentum J and nuclear angular momentum I are only partially decoupled. As a result of the mixing of wavefunctions of different hyperfine states, we observe a nonlinear Zeeman effect for each sublevel, a substantial modification of the transition probabilities between different magnetic sublevels, and the appearance of transitions that are strictly forbidden in the absence of the magnetic field. For the case of right- and left- handed circularly polarized laser excitation, the fluorescence spectra differs qualitatively. Well pronounced magnetic field induced circular dichroism is observed. These observations are explained with a standard approach that describes the partial decoupling of I and J states

MicroRNA regulation of endothelial TREX1 reprograms the tumour microenvironment

Rather than targeting tumour cells directly, elements of the tumour microenvironment can be modulated to sensitize tumours to the effects of therapy. Here we report a unique mechanism by which ectopic microRNA-103 can manipulate tumour-associated endothelial cells to enhance tumour cell death. Using gain-and-loss of function approaches, we show that miR-103 exacerbates DNA damage and inhibits angiogenesis in vitro and in vivo. Local, systemic or vascular-targeted delivery of miR-103 in tumour-bearing mice decreased angiogenesis and tumour growth. Mechanistically, miR-103 regulation of its target gene TREX1 in endothelial cells governs the secretion of pro-inflammatory cytokines into the tumour microenvironment. Our data suggest that this inflammatory milieu may potentiate tumour cell death by supporting immune activation and inducing tumour expression of Fas and TRAIL receptors. Our findings reveal miR-mediated crosstalk between vasculature and tumour cells that can be exploited to improve the efficacy of chemotherapy and radiation.United States. National Institutes of Health (R00HL112962)United States. National Institutes of Health (R01 HL57900)Oregon Health & Science University. Knight Cancer Institute (2015-Dive-Knight-01