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

Agarose-based structured optical fibre

Biocompatible and resorbable optical fibres emerge as promising technologies for in vivo applications like imaging, light delivery for phototherapy and optogenetics, and localised drug-delivery, as well as for biochemical sensing, wherein the probe can be implanted and then completely absorbed by the organism. Biodegradable waveguides based on glasses, hydrogels, and silk have been reported, but most of these devices rely on complex fabrication procedures. In this sense, this paper proposes a novel structured optical fibre made of agarose, a transparent, edible material used in culture media and tissue engineering. The fibre is obtained by pouring food-grade agar into a mould with stacked rods, forming a solid core surrounded by air holes in which the refractive index and fibre geometry can be tailored by choosing the agarose solution composition and mould design, respectively. Besides exhibiting practical transmittance at 633 nm in relation to other hydrogel waveguides, the fibre is also validated for chemical sensing either by detecting volume changes due to agar swelling/dehydration or modulating the transmitted light by inserting fluids into the air holes. Therefore, the proposed agarose-based structured optical fibre is an easy-to-fabricate, versatile technology with possible applications for medical imaging and in vivo biochemical sensing101CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPNão temNão tem2017/25666-

Biochemical Characterization of Anopheles gambiae SRPN6, a Malaria Parasite Invasion Marker in Mosquitoes

Serine proteinase inhibitors of the serpin family are well known as negative regulators of hemostasis, thrombolysis and innate immune responses. Additionally, non-inhibitory serpins serve functions as chaperones, hormone transporters, or anti-angiogenic factors. In the African malaria mosquito, Anopheles gambiae s.s., at least three serpins (SRPNs) are implicated in the innate immune response against malaria parasites. Based on reverse genetic and cell biological analyses, AgSRPN6 limits parasite numbers and transmission and has been postulated to control melanization and complement function in mosquitoes. This study aimed to characterize AgSRPN6 biophysically and determine its biochemical mode of action. The structure model of AgSRPN6, as predicted by I-Tasser showed the protein in the native serpin fold, with three central β-sheets, nine surrounding α-helices, and a protruding reactive center loop. This structure is in agreement with biophysical and functional data obtained from recombinant (r) AgSRPN6, produced in Escherichia coli. The physical properties of purified rAgSRPN6 were investigated by means of analytical ultracentrifugation, circular dichroism, and differential scanning calorimetry tools. The recombinant protein exists predominantly as a monomer in solution, is composed of a mixture of α-helices and β-sheets, and has a mid-point unfolding temperature of 56°C. Recombinant AgSRPN6 strongly inhibited porcine pancreatic kallikrein and to a lesser extent bovine pancreatic trypsin in vitro. Furthermore, rAgSRPN6 formed inhibitory, SDS-stable, higher molecular weight complexes with prophenoloxidase-activating proteinase (PAP)1, PAP3, and Hemolymph protein (HP)6, which are required for melanization in the lepidopteran model organism, Manduca sexta. Taken together, our results strongly suggest that AgSRPN6 takes on a native serpin fold and is an inhibitor of trypsin-like serine proteinases.This work was supported by the National Institutes of Health through 3P20RR017708-07S1 and P20RR017686 sub-awards and 1R01AI095842 to K.M. This is contribution 12-098-J from the Kansas Agricultural Experiment Station

Design of 11-Residue Peptides with Unusual Biophysical Properties: Induced Secondary Structure in the Absence of Water

AbstractA series of oligopeptides with β-forming and adhesive properties, were synthesized and analyzed for adhesion shear strength, secondary structure, and association properties. The sequences contained related hydrophobic core segments varying in length from 5 to 12 residues and flanked by di- or tri-lysine segments. Three remarkable peptides consisting of just 11 residues with hydrophobic core sequences of FLIVI, IGSII, and IVIGS flanked by three lysine residues gave the highest dry adhesion shear strength and displayed unusual biophysical properties in the presence and absence of water. KKKFLIVIKKK had its highest adhesion strength at 2% (w/v) at pH 12.0 and showed the highest adhesion strength after exposure to water (water resistance). Both KKKIGSIIKKK and KKKIVIGSKKK, at 4% (w/v) at pH 12.0, displayed nearly identical dry shear strength values to that with the FLIVI core sequence. The peptide with IGSII core, however, displayed a lower water resistance and the latter, IVIGS, showed no water resistance, completely delaminating upon soaking in water. These are the smallest peptides with adhesive properties reported to date and show remarkable adhesion strength even at lower concentrations of 0.2% (w/v), which corresponds to 1.6mM. The FLIVI containing peptide adopted a β-sheet secondary structure in water while the IGSII- and IVIGS-containing sequences folded similarly only in the absence of water. Analytical ultracentrifugation studies showed that when the FLIVI sequence adopts β-structure in aqueous solution, it associates into a large molecular weight assembly. The random coils of IGSII and IVIGS showed no tendency to associate at any pH

The First Focused Hard X-ray Images of the Sun with NuSTAR

We present results from the the first campaign of dedicated solar observations undertaken by the \textit{Nuclear Spectroscopic Telescope ARray} ({\em NuSTAR}) hard X-ray telescope. Designed as an astrophysics mission, {\em NuSTAR} nonetheless has the capability of directly imaging the Sun at hard X-ray energies ($>$3~keV) with an increase in sensitivity of at least two magnitude compared to current non-focusing telescopes. In this paper we describe the scientific areas where \textit{NuSTAR} will make major improvements on existing solar measurements. We report on the techniques used to observe the Sun with \textit{NuSTAR}, their limitations and complications, and the procedures developed to optimize solar data quality derived from our experience with the initial solar observations. These first observations are briefly described, including the measurement of the Fe K-shell lines in a decaying X-class flare, hard X-ray emission from high in the solar corona, and full-disk hard X-ray images of the Sun.Comment: 11 pages, accepted to Ap

Hibernoma of the axillary region: a rare benign adipocytic tumor

Hibernoma is a rare benign tumor considered to arise from remnants of fetal brown adipose tissue. It tends to occur in sites where brown fat persists beyond fetal life, such as the interscapular region, but can occur in sites where brown fat is usually absent in adults. Clinicallywell, hibernomas are slow-growing, asymptomatic tumors. However, unlike lipomas, MRI findings sometimes mislead clinicians to diagnose a malignant neoplasm. We describe a 63-year-old male with an axillary hibernoma involving the brachial neurovascular bundles and mimicking a well-differentiated liposarcoma, from which it should be distinguished

Characterization of Redlen CZT detectors for hard x-ray astronomy

We present the results of ongoing characterization of Cadmium Zinc Telluride (CZT) semiconductors produced by Redlen Technologies. In particular we hope to determine their viability for future X-ray astronomy missions such as the High Energy X-ray Probe (HEX-P). The fully fabricated hybrid detectors consist of CZT crystals with a collecting area of 2 cm × 2 cm and thickness of 3 mm mounted on a custom pixelated ASIC originally designed for the Nuclear Spectroscopic Telescope Array (NuSTAR) mission, which launched in 2012. We present the results of inter-pixel conductance and leakage current tests as well as spectral characterization using an 241Am source. Although further calibration and testing is necessary to determine the capabilities of these detectors, preliminary results indicate that Redlen CZT will be able to achieve spectral resolution and noise levels comparable to those of the CZT detectors currently in use aboard NuSTAR

Precision test of gauge/gravity duality in D0-brane matrix model at low temperature

We test the gauge/gravity duality between the matrix model and type IIA string theory at low temperatures with unprecedented accuracy. To this end, we perform lattice Monte Carlo simulations of the Berenstein-Maldacena-Nastase (BMN) matrix model, which is the one-parameter deformation of the Banks-Fischler-Shenker-Susskind (BFSS) matrix model, taking both the large $N$ and continuum limits. We leverage the fact that sufficiently small flux parameters in the BMN matrix model have a negligible impact on the energy of the system while stabilizing the flat directions so that simulations at smaller $N$ than in the BFSS matrix model are possible. Hence, we can perform a precision measurement of the large $N$ continuum energy at the lowest temperatures to date. The energy is in perfect agreement with supergravity predictions including estimations of $\alpha'$-corrections from previous simulations. At the lowest temperature where we can simulate efficiently ($T=0.25\lambda^{1/3}$, where $\lambda$ is the 't Hooft coupling), the difference in energy to the pure supergravity prediction is less than $10\%$. Furthermore, we can extract the coefficient of the $1/N^4$ corrections at a fixed temperature with good accuracy, which was previously unknown.Comment: 33 pages, 16 figures. Data are available upon request to Stratos Pateloudi

NuSTAR Observations of X-Ray Binaries

As of 2014 August, the Nuclear Spectroscopic Telescope Array (NuSTAR) had observed ~30 X-ray binaries either as part of the planned program, as targets of opportunity, or for instrument calibration. The main science goals for the observations include probing the inner part of the accretion disk and constraining black hole spins via reflection components, providing the first observations of hard X-ray emission from quiescent Low Mass X-ray Binaries (LMXBs), measuring cyclotron lines from accreting pulsars, and studying type I X-ray bursts from neutron stars. Here, we describe the science objectives in more depth and give an overview of the NuSTAR observations that have been carried out to achieve the objectives. These include observation of four "IGR" High Mass X-ray Binaries (HMXBs) discovered by INTEGRAL. We also summarize the results that have been obtained and their implications. Among the IGR HMXBs, we focus on the discovery of a cyclotron line in the spectrum of IGR J17544-2619

The Nuclear Spectroscopic Telescope Array (NuSTAR)

The Nuclear Spectroscopic Telescope Array (NuSTAR) is a NASA Small Explorer mission that will carry the first focusing hard X-ray (5 -- 80 keV) telescope to orbit. NuSTAR will offer a factor 50 -- 100 sensitivity improvement compared to previous collimated or coded mask imagers that have operated in this energy band. In addition, NuSTAR provides sub-arcminute imaging with good spectral resolution over a 12-arcminute field of view. After launch, NuSTAR will carry out a two-year primary science mission that focuses on four key programs: studying the evolution of massive black holes through surveys carried out in fields with excellent multiwavelength coverage, understanding the population of compact objects and the nature of the massive black hole in the center of the Milky Way, constraining explosion dynamics and nucleosynthesis in supernovae, and probing the nature of particle acceleration in relativistic jets in active galactic nuclei. A number of additional observations will be included in the primary mission, and a guest observer program will be proposed for an extended mission to expand the range of scientific targets. The payload consists of two co-aligned depth-graded multilayer coated grazing incidence optics focused onto solid state CdZnTe pixel detectors. To be launched in early 2012 on a Pegasus rocket into a low-inclination Earth orbit. Data will be publicly available at GSFC's High Energy Astrophysics Science Archive Research Center (HEASARC) following validation at the science operations center located at Caltech.Comment: 9 pages, 5 figures, to appear in Proceedings of the SPIE, Space Telescopes and Instrumentation 2010: Ultraviolet to Gamma Ra