Precursor ion scanning for detection and structural characterization of heterogeneous glycopeptide mixtures

AbstractThe structure of N-linked glycans is determined by a complex, anabolic, intracellular pathway but the exact role of individual glycans is not always clear. Characterization of carbohydrates attached to glycoproteins is essential to aid understanding of this complex area of biology. Specific mass spectral detection of glycopeptides from protein digests may be achieved by on-line HPLC-MS, with selected ion monitoring (SIM) for diagnostic product ions generated by cone voltage fragmentation, or by precursor ion scanning for terminal saccharide product ions, which can yield the same information more rapidly. When glycosylation is heterogeneous, however, these approaches can result in spectra that are complex and poorly resolved. We have developed methodology, based around precursor ion scanning for ions of high m/z, that allows site specific detection and structural characterization of glycans at high sensitivity and resolution. These methods have been developed using the standard glycoprotein, fetuin, and subsequently applied to the analysis of the N-linked glycans attached to the scrapie-associated prion protein, PrPSc. These glycans are highly heterogeneous and over 30 structures have been identified and characterized site specifically. Product ion spectra have been obtained on many glycopeptides confirming structure assignments. The glycans are highly fucosylated and carry Lewis X or sialyl Lewis X epitopes and the structures are in-line with previous results. [Abbreviations: Hex–Hexose, C6H12O6 carbohydrates, including mannnose and galactose; HexNAc—N-acetylhexosamine, C8H15NO6 carbohydrates, including N-acetylglucosamine and N-acetylgalactosamine; GlcNAc—N-acetylglucosamine; GalNAc—N-acetylgalactosamine; Fuc–Fucose; NeuAC—N-acetylneuraminic acid or sialic acid; TSE—Transmissible Spongiform Encephalopathy.

A new code for parameter estimation in searches for gravitational waves from known pulsars

We describe the consistency testing of a new code for gravitational wave signal parameter estimation in known pulsar searches. The code uses an implementation of nested sampling to explore the likelihood volume. Using fake signals and simulated noise we compare this to a previous code that calculated the signal parameter posterior distributions on both a grid and using a crude Markov chain Monte Carlo (MCMC) method. We define a new parameterisation of two orientation angles of neutron stars used in the signal model (the initial phase and polarisation angle), which breaks a degeneracy between them and allows more efficient exploration of those parameters. Finally, we briefly describe potential areas for further study and the uses of this code in the future.Comment: Accepted for proceedings of Amaldi 9 meetin

Rotorcraft convertible engine study

The objective of the Rotorcraft Convertible Engine Study was to define future research and technology effort required for commercial development by 1988 of convertible fan/shaft gas turbine engines for unconventional rotorcraft transports. Two rotorcraft and their respective missions were defined: a Fold Tilt Rotor aircraft and an Advancing Blade Concept (ABC) rotorcraft. Sensitivity studies were conducted with these rotorcraft to determine parametrically the influence of propulsion characteristics on aircraft size, mission fuel requirements, and direct operating costs (DOC). The two rotorcraft were flown with conventional propulsion systems (separate lift/cruise engines) and with convertible propulsion systems to determine the benefits to be derived from convertible engines. Trade-off studies were conducted to determine the optimum engine cycle and staging arrangement for a convertible engine. Advanced technology options applicable to convertible engines were studied. Research and technology programs were identified which would ensure technology readiness for commercial development of convertible engines by 1988

On the Interface Between LENS® Deposited Stainless Steel 304L Repair Geometry and Cast or Machined Components

Laser Engineered Net Shaping™ (LENS®) is being evaluated for use as a metal component repair/modification process. A component of the evaluation is to better understand the characteristics of the interface between LENS deposited material and the substrate on which it is deposited. A processing and metallurgical evaluation was made on LENS processed material fabricated for component qualification tests. A process parameter evaluation was used to determine optimum build parameters and these parameters were used in the fabrication of tensile test specimens to study the characteristics of the interface between LENS deposited material and several types of substrates. Analyses of the interface included mechanical properties, microstructure, and metallurgical integrity. Test samples were determined for a variety of geometric configurations associated with interfaces between LENS deposited material and both wrought base material or previously deposited LENS material. Thirteen different interface configurations were fabricated for evaluation representing a spectrum of deposition conditions from complete part build, to hybrid substrate-LENS builds, to repair builds for damaged or re-designed housings. Good mechanical properties and full density were observed for all configurations. When tested to failure, fracture occurred by ductile microvoid coalescence. The repair and hybrid interfaces showed the same metallurgical integrity as, and had properties similar to, monolithic LENS deposits.Mechanical Engineerin

Analytical and structural studies of acacia polysaccharides

An analytical study of eleven Acacia gums from the subseries Juliflorae of the series Phyllodineae showed them to be more proteinaceous, more acidic and more viscous, with higher methoxyl contents and higher molecular weights but with lower proportions of rhamnose and arabinose than the majority of Acacia gums studied so far.A structural study of the gum from Acacia auriculiformis involving hydrolysis, Smith- degradation and methylation studies, revealed a ß -1, 3 - linked galactose backbone with side chains of ß -1, 6 - linked galactose. Glucuronic acid was present as end-groups, linked ß-1, 6 to galactose: 4-Ω-methyl glucuronic acid was present as end - groups, α -1, 4 - linked to galactose. Arabinose was present in short, ß-1, 3- linked chains, while rhamnose was present as end-group. Proteinaceous material was attached to the molecule by periodate- resistant linkages.The use of carbon 13-nuclear magnetic resonance spectroscopy for the study of gum exudates was examined. Spectra were obtained for the gum from Acacia auriculiformis and its degradation products, despite their high molecular weight and chemical complexity. The spectra substantiated the findings of the structural study

Float zone experiments in space

The molten zone/freezing crystal interface system and all the mechanisms were examined. If Marangoni convection produces oscillatory flows in the float zone of semiconductor materials, such as silicon, then it is unlikely that superior quality crystals can be grown in space using this process. The major goals were: (1) to determine the conditions for the onset of Marangoni flows in molten tin, a model system for low Prandtl number molten semiconductor materials; (2) to determine whether the flows can be suppressed by a thin oxide layer; and (3) based on experimental and mathematical analysis, to predict whether oscillatory flows will occur in the float zone silicon geometry in space, and if so, could it be suppressed by thin oxide or nitride films. Techniques were developed to analyze molten tin surfaces in a UHV system in a disk float zone geometry to minimize buoyancy flows. The critical Marangoni number for onset of oscillatory flows was determined to be greater than 4300 on atomically clean molten tin surfaces

Calcium binding activity of the epidermal growth factor-like domains of the apicomplexan microneme protein EtMIC4

Microneme proteins are secreted from apicomplexan parasites during invasion of host cells and they play crucial roles in parasite-host cell adhesion. EtMIC4 is a 240 kDa transmembrane protein from Eimeria tenella that contains 31 tandemly arranged epidermal growth factor (EGF), like repeats within its extracellular domain. The majority of these repeats have calcium binding (cb) consensus sequences. Little is known about cbEGFs in apicomplexan parasites but their presence in microneme proteins suggests that they may contribute to parasite-host interactions. To investigate the potential role of cbEGFs we have expressed and correctly refolded a cbEGF triplet from EtMIC4 (cbEGF7-9) and demonstrated that this triplet binds calcium. Circular dichroism spectroscopic analysis of cbEGF7-9 demonstrates that the molecule undergoes a gradual change in conformation with increasing levels of calcium. In the presence of calcium, the triplet becomes resistant to proteolytic degradation by a variety of proteases, a characteristic feature of cbEGF repeats from higher eukaryotic proteins, such as fibrillin, suggesting that calcium binding induces the formation of a rigid conformation. Moreover, mass spectrometric mapping of the cleavage sites that are protected by calcium shows that these sites are located both close to and distant from the calcium binding sites, indicating that protection is not due to steric hindrance by calcium ions, but rather due to the overall conformation adopted by the triplet in the presence of calcium. Thus, the tandemly-arranged cbEGF repeats within EtMIC4 provide a mechanism whereby, in the calcium-rich extracellular environment, the molecule could adopt a protease-resistant, rigid structure that could favour its interaction with host cell ligands

Is the transition redshift a new cosmological number?

Observations from Supernovae Type Ia (SNe Ia) provided strong evidence for an expanding accelerating Universe at intermediate redshifts. This means that the Universe underwent a transition from deceleration to acceleration phases at a transition redshift $z_t$ of the order unity whose value in principle depends on the cosmology as well as on the assumed gravitational theory. Since cosmological accelerating models endowed with a transition redshift are extremely degenerated, in principle, it is interesting to know whether the value of $z_t$ itself can be observationally used as a new cosmic discriminator. After a brief discussion of the potential dynamic role played by the transition redshift, it is argued that future observations combining SNe Ia, the line-of-sight (or "radial") baryon acoustic oscillations, the differential age of galaxies, as well as the redshift drift of the spectral lines may tightly constrain $z_t$, thereby helping to narrow the parameter space for the most realistic models describing the accelerating Universe.Comment: 12 pages, 5 figures. Some discussions about how to estimate the transition redshift have been added. New data by Planck and H(z) data have been mentioned. New references have been adde

Acute Liver Failure Secondary to Hemophagocytic Lymphohistiocytosis during Pregnancy.

Hemophagocytic lymphohistiocytosis (HLH) is a syndrome of excessive immune activation that mimics and occurs with other systemic diseases. A 35-year-old female presented with signs of viral illness at 13 weeks of pregnancy and progressed to acute liver failure (ALF). We discuss the diagnosis of HLH and Kikuchi-Fujimoto (KF) lymphadenitis in the context of pregnancy and ALF. HLH may respond to comorbid disease-specific therapy, and more toxic treatment can be avoided