Cryo-electron tomography of Marburg Virus particles and their morphogenesis within infected cells

Several major human pathogens, including the filoviruses, paramyxoviruses, and rhabdoviruses, package their single-stranded RNA genomes within helical nucleocapsids, which bud through the plasma membrane of the infected cell to release enveloped virions. The virions are often heterogeneous in shape, which makes it difficult to study their structure and assembly mechanisms. We have applied cryo-electron tomography and sub-tomogram averaging methods to derive structures of Marburg virus, a highly pathogenic filovirus, both after release and during assembly within infected cells. The data demonstrate the potential of cryo-electron tomography methods to derive detailed structural information for intermediate steps in biological pathways within intact cells. We describe the location and arrangement of the viral proteins within the virion. We show that the N-terminal domain of the nucleoprotein contains the minimal assembly determinants for a helical nucleocapsid with variable number of proteins per turn. Lobes protruding from alternate interfaces between each nucleoprotein are formed by the C-terminal domain of the nucleoprotein, together with viral proteins VP24 and VP35. Each nucleoprotein packages six RNA bases. The nucleocapsid interacts in an unusual, flexible "Velcro-like" manner with the viral matrix protein VP40. Determination of the structures of assembly intermediates showed that the nucleocapsid has a defined orientation during transport and budding. Together the data show striking architectural homology between the nucleocapsid helix of rhabdoviruses and filoviruses, but unexpected, fundamental differences in the mechanisms by which the nucleocapsids are then assembled together with matrix proteins and initiate membrane envelopment to release infectious virions, suggesting that the viruses have evolved different solutions to these conserved assembly steps.

Analytical electron microscope investigation of boron carbide microstructures at elevated temperatures

The microstructures of hot-pressed B4C were monitored during in situ heating experiments from room temperature to 1000C by analytical electron microscopy (AEM). Variations in the microstructure of B4C were not observed. However, during heating, secondary phases formed in voids and on the surfaces of the specimen.

Thin-film analyses of silicate standards at 200 kv : the effect of temperature on element loss

Preliminary data is presented on a detailed statistical analysis of k-factor determination for a single class of minerals (amphiboles) which contain a wide range of element concentrations. These amphiboles are homogeneous, contain few (if any) subsolidus microstructures and can be readily prepared for thin film analysis. In previous studies, element loss during the period of irradiation has been assumed negligible for the determination of k-factors. Since this phenomena may be significant for certain mineral systems, we also report on the effect of temperature on k-factor determination for various elements using small probe sizes (approx.20 nm).

Electron paramagnetic resonance, optical absorption and Raman spectral studies on a pyrite/chalcopyrite mineral

Pyrite and chalcopyrite mineral samples from Mangampet barite mine, Kadapa, Andhra Pradesh, India are used in the present study. XRD data indicate that the pyrite mineral has a face centered cubic lattice structure with lattice constant 5.4179 Å. Also it possesses an average particle size of 91.9 nm. An EPR study on the powdered samples confirms the presence of iron in pyrite and iron and Mn(II) in chalcopyrite. The optical absorption spectrum of chalcopyrite indicates presence of copper which is in a distorted octahedral environment. NIR results confirm the presence of water fundamentals and Raman spectrum reveals the presence of water and sulfate ions.

Influence of electrolyte cations on electron transport and electron transfer in dye-sensitized solar cells

The influence of different electrolyte cations ((Li+, Na+, Mg2+, tetrabutyl ammonium (TBA+)) on the TiO2 conduction band energy (Ec) the effective electron lifetime (τn), and the effective electron diffusion coefficient (Dn) in dye-sensitized solar cells (DSCs) was studied quantitatively. The separation between Ec and the redox Fermi level, EF,redox, was found to decrease as the charge/radius ratio of the cations increased. Ec in the Mg2+ electrolyte was found to be 170 meV lower than that in the Na+ electrolyte and 400 meV lower than that in the TBA+ electrolyte. Comparison of Dn and τn in the different electrolytes was carried out by using the trapped electron concentration as a measure of the energy difference between Ec and the quasi-Fermi level, nEF, under different illumination levels. Plots of Dn as a function of the trapped electron density, nt, were found to be relatively insensitive to the electrolyte cation, indicating that the density and energetic distribution of electron traps in TiO2 are similar in all of the electrolytes studied. By contrast, plots of τn versus nt for the different cations showed that the rate of electron back reaction is more than an order of magnitude faster in the TBA+ electrolyte compared with the Na+ and Li+ electrolytes. The electron diffusion lengths in the different electrolytes followed the sequence of Na+ > Li+ > Mg2+ > TBA+. The trends observed in the AM 1.5 current–voltage characteristics of the DSCs are rationalized on the basis of the conduction band shifts and changes in electron lifetime.

Fast scanning electron microscope (FSEM)

High magnification and large depth of field with a temporal resolution of less than 100 microseconds are possible using the present invention which combines a linear electron beam produced by a tungsten filament from an SX-40A Scanning Electron Microscope (SEM), a magnetic deflection coil with lower inductance resulting from reducing the number of turns of the saddle-coil wires, while increasing the diameter of the wires, a fast scintillator, photomultiplier tube, photomultiplier tube base, and signal amplifiers and a high speed data acquisition system which allows for a scan rate of 381 frames per second and 256.times.128 pixel density in the SEM image at a data acquisition rate of 25 MHz. The data acquisition and scan position are fully coordinated. A digitizer and a digital waveform generator which generates the sweep signals to the scan coils run off the same clock to acquire the signal in real-time.

Elemental characterisation and detection of heavy metals in house dusts collected from sites in South-East Queensland, Australia

Knowledge of the elements present in house dusts is important in understanding potential health effects on humans. In this study, dust samples collected from 10 houses in south-east Queensland have been analysed by scanning electron microscopy and X-ray microanalysis to measure the inorganic element compositions and to investigate the form of heavy metals in the dusts. The overall analytical results were then used to discriminate between different localities using chemometric techniques. The relative amounts of elements, particularly of Si, Ca, and Fe, varied between size fractions and between different locations for the same size fraction. By analysing individual small particles, many other constituents were identified including Ti, Cr, Mn, Ni, Cu, Zn, Ba, Ag, W, Au, Hg, Pb, Bi, La and Ce. The heavy metals were mostly concentrated in small particles in the smaller size fractions, which allowed detection by particle analysis, though their average concentrations were very low.

High resolution transmission electron microscopy of the Murchison Carbonaceous Chondrite and the Kenna Ureilite

Fine-grained matrices in carbonaceous chondrites and small, micron-sized inclusions in achondrites can be characterized effectively using high resolution transmission electron micro­scopy (HRTEM).

Distinguishing new science from calibration effects in the electron-volt neutron spectrometer VESUVIO at ISIS

The "standard" procedure for calibrating the Vesuvio eV neutron spectrometer at the ISIS neutron source, forming the basis for data analysis over at least the last decade, was recently documented in considerable detail by the instrument’s scientists. Additionally, we recently derived analytic expressions of the sensitivity of recoil peak positions with respect to fight-path parameters and presented neutron–proton scattering results that together called in to question the validity of the "standard" calibration. These investigations should contribute significantly to the assessment of the experimental results obtained with Vesuvio. Here we present new results of neutron–deuteron scattering from D2 in the backscattering angular range (theata > 90 degrees) which are accompanied by a striking energy increase that violates the Impulse Approximation, thus leading unequivocally the following dilemma: (A) either the "standard" calibration is correct and then the experimental results represent a novel quantum dynamical effect of D which stands in blatant contradiction of conventional theoretical expectations; (B) or the present "standard" calibration procedure is seriously deficient and leads to artificial outcomes. For Case(A), we allude to the topic of attosecond quantumdynamical phenomena and our recent neutron scattering experiments from H2 molecules. For Case(B),some suggestions as to how the "standard" calibration could be considerably improved are made.

Scanning electron microscopy of condensation related minerals : the Bjurbole meteorite

Filamentary single crystals, blades, sheets, euhedral crystals and powders may form by vapor phase condensation depending on the supersauration conditions in the vapor with respect to the condensing species [1]. Filamentary crystal growth requires the operation of an axial screw dislocation [2]. A Vapor-Liquid-Solid (VLS) mechanism may also produce filamentary single crystals, ribbons and blades. The latter two morphologies are typically twinned. Crystals grown by this mechanism do not require the presence of an axial screw dislocation. Impurities may either promote or inhibit crystal growth [3]. The VLS mechanism allows crystals to grow at small supersaturation of the vapor. Thin enstatite blades, ribbons and sheets have been observed in chondritic porous Interplanetary Dust Partics (IDP's) [4, 5]. The requisite screw dislocation for vapor phase condensation [1] has been observed in these enstatite blades [4]. Bradley et al. [4] suggest that these crystals are primary vapor phase condensates which could have formed either in the solar nebula or in presolar environments. These observations [4,5] are significant in that they may provide a demonstrable link to theoretical predictions: viz. that in the primordial solar nebula filamentary condensates could cluster into 'lint balls' and form the predecessors to comets [6].

Electron microscopy of stratospheric particles

A recent NASA program to collect stratospheric dust particles using high-flying WB57 aircraft has made available many more potential candidates for the study of extraterrestrial materials. This preliminary report provides an interpretation of the types of particles returned from one flag (W7017) collected in August, 1981 using a subset of 81 allocated particles. This particular collection period is after the Mt. St. Helen's eruptions. Therefore, the flag may contain significant quantities of volcanic debris in addition to the expected terrestrial contaminants [1]. All particles were mounted on nucleopore filters and have been examined using a modified JEOL100CX analytical electron microscope. For most of the particles, X-ray energy dispersive spectra and images were obtained at 40kV on samples which have not received any conductive coating. However, in order to improve resolution (to ~30A) some images are recorded at 100kV. In addition, 16 samples have been coated with a thin layer (<50A) of Au/Pd.

Electron microscopy of micron-sized lunar soil

We have performed electron-microscopic analysis on 0.5-1.0µm grains in order to study radiation damage by the solar-wind. We are reporting some interesting results we have found in monomineralic grains from core sample 15010,1130. This is a submature soil which has been studied for rare gas abundance and ferromagnetic resonance by (1) and modal petrology by (2).

The Use of a Nitroxide Probe in DMSO to Capture Radicals in Particulate Pollution

A profluorescent nitroxide was used to evaluate the oxidative potential of pollution derived from a compression ignition engine fuelled with biodiesel. The reaction products responsible for the observed fluorescence increase when a DMSO solution of nitroxide was exposed to biodiesel exhaust were determined by using HPLC/MS. The main fluorescent species was identified as a methanesulfonamide adduct arising from the reaction of the nitroxide with DMSO-derived sulfoxyl radicals.

Composition and morphology of particle emissions from in-use aircraft during takeoff and landing

In order to provide realistic data for air pollution inventories and source apportionment at airports, the morphology and composition of ultrafine particles (UFP) in aircraft engine exhaust were measured and characterized. For this purpose, two independent measurement techniques were employed to collect emissions during normal takeoff and landing operations at Brisbane Airport, Australia. PM1 emissions in the airfield were collected on filters and analyzed using the particle-induced X-ray emission (PIXE) technique. Morphological and compositional analyses of individual ultrafine particles in aircraft plumes were performed on silicon nitride membrane grids using transmission electron microscopy (TEM) combined with energy-dispersive X-ray microanalysis (EDX). TEM results showed that the deposited particles were in the range of 5 to 100 nm in diameter, had semisolid spherical shapes and were dominant in the nucleation mode (18 – 20 nm). The EDX analysis showed the main elements in the nucleation particles were C, O, S and Cl. The PIXE analysis of the airfield samples was generally in agreement with the EDX in detecting S, Cl, K, Fe and Si in the particles. The results of this study provide important scientific information on the toxicity of aircraft exhaust and their impact on local air quality.

Calibration of the electron volt spectrometer, a deep inelastic neutron scattering spectrometer at the ISIS pulsed neutron spallation source

The electron Volt Spectrometer (eVS) is an inverse geometry filter difference spectrometer that has been optimised to measure the single atom properties of condensed matter systems using a technique known as Neutron Compton Scattering (NCS) or Deep Inelastic Neutron Scattering (DINS). The spectrometer utilises the high flux of epithermal neutrons that are produced by the ISIS neutron spallation source enabling the direct measurement of atomic momentum distributions and ground state kinetic energies. In this paper the procedure that is used to calibrate the spectrometer is described. This includes details of the method used to determine detector positions and neutron flight path lengths as well as the determination of the instrument resolution. Examples of measurements on 3 different samples are shown, ZrH2, 4He and Sn which show the self-consistency of the calibration procedure.