Physicochemical studies on the gelation of hen's egg yolk: Delipidation of yolk plasma by treatment with phospholipase-C and extraction with solvents

A method for the delipidation of egg yolk plasma using phospholipase-C, n-heptane, and 1-butanol has been described. An aggregating protein fraction and a soluble protein fraction were separated by the action of phospholipase-C. The aggregating protein fraction freed of most of the lipids by treatment with n-heptane and 1-butanol was shown to be the apolipoproteins of yolk plasma, whereas the soluble proteins were identified as the livetins. Carbohydrate and the N-terminal amino acid analysis of these protein fractions are reported. A comparison of these protein fractions with the corresponding fractions obtained by formic acid delipidation of yolk plasma has been made. The gelation of yolk plasma by the action of phospholipase-C has been interpreted as an aggregation of lipoproteins caused by ionic interactions. The role of lecithin in maintaining the structural integrity of lipoproteins has been discussed.

Glycoprotein Interactions in the Assembly of Hantaviruses

Hantaviruses are one of the five genera of the vector-borne virus family Bunyaviridae. While other members of the family are transmitted via arthropods, hantaviruses are carried and transmitted by rodents and insectivores. Occasional transmission to humans occurs via inhalation of aerosolized rodent excreta. When transmitted to man hantaviruses cause hemorrhagic fever with renal syndrome (HFRS, in Eurasia, mortality ~10%) and hantavirus cardiopulmonary syndrome (HCPS, in the Americas, mortality ~40%). The single-stranded, negative-sense RNA genome of hantaviruses is in segments S, M and L that respectively encode for nucleocapsid (N), glycoproteins Gn and Gc, and RNA-dependent RNA-polymerase (RdRp or L protein). The genome segments, encapsidated by N protein to form ribonucleoprotein (RNP), are enclosed inside a lipid envelope decorated by spikes formed of Gn and Gc. The focus of this study was to understand the mechanisms and interactions through which the virion is formed and maintained. We observed that when extracted from virions both Gn and Gc favor homo- over hetero-oligomerization. The minimal glycoprotein complexes extracted from virion by detergent were observed, by using ultracentrifugation and gel filtration, to be tetrameric Gn and homodimeric Gc. These results led us to suggest a model where tetrameric Gn complexes are interconnected through homodimeric Gc units to form the grid-like surface architecture described for hantaviruses. This model was found to correlate with the three-dimensional (3D) reconstruction of virion surface created using cryo-electron tomography (cryo-ET). The 3D-density map showed the spike complex formed of Gn and Gc to be 10 nm high and to display a four-fold symmetry with dimensions of 15 nm times 15 nm. This unique square-shaped complex on a roughly round virion creates a hitch for the assembly, since a sphere cannot be broken into rectangles. Thus additional interactions are likely required for the virion assembly. In cryo-ET we observed that the RNP makes occasional contacts to the viral membrane, suggesting an interaction between the spike and RNP. We were able to demonstrate this interaction using various techniques, and showed that both Gn and Gc contribute to the interaction. This led us to suggest that in addition to the interactions between Gn and Gc, also the interaction between spike and RNP is required for assembly. We found galectin-3 binding protein (referred to as 90K) to co-purify with the virions and showed an interaction between 90K and the virion. Analysis of plasma samples taken from patients hospitalized for Puumala virus infection showed increased concentrations of 90K in the acute phase and the increased 90K level was found to correlate with several parameters that reflect the severity of acute HFRS. The results of these studies confirmed, but also challenged some of the dogmas on the structure and assembly of hantaviruses. We confirmed that Gn and RNP do interact, as long assumed. On the other hand we demonstrated that the glycoproteins Gn and Gc exist as homo-oligomers or appear in large hetero-oligomeric complexes, rather than form primarily heterodimers as was previously assumed. This work provided new insight into the structure and assembly of hantaviruses.

Quark and gluon distribution functions in a viscous quark-gluon plasma medium and dilepton production via q(q)over-bar annihilation

Viscous modifications to the thermal distributions of quark-antiquarks and gluons have been studied in a quasiparticle description of the quark-gluon-plasma medium created in relativistic heavy-ion collision experiments. The model is described in terms of quasipartons that encode the hot QCD medium effects in their respective effective fugacities. Both shear and bulk viscosities have been taken in to account in the analysis, and the modifications to thermal distributions have been obtained by modifying the energy-momentum tensor in view of the nontrivial dispersion relations for the gluons and quarks. The interactions encoded in the equation of state induce significant modifications to the thermal distributions. As an implication, the dilepton production rate in the q (q) over bar annihilation process has been investigated. The equation of state is found to have a significant impact on the dilepton production rate along with the viscosities.

Interactions of a femtosecond intense laser with rare gas clusters in a dense jet\

A study on the interactions of high intensity (similar to 10(16) W/cm(2)) femtosecond laser pulses with rare gas clusters in a dense jet is performed. Energy absorption by Ar and Xe clusters is measured and it can be as high as 90%. Very energetic ions produced in the laser interaction with a dense cluster jet are detected by time-of-flight spectrometry and the maximum ion energy of Xe is up to 1.3 MeV. The average ion energies are found to increase with increasing cluster size and get saturated gradually. The average ion energies also show a strong directionality and the average ion energy in the direction parallel to the laser polarization vector is 40% higher than that perpendicular to it. The findings are discussed in terms of a model of charge-dependent ion acceleration.

Studies on morphogenesis and cellular interactions of Rana grylio virus in an infected fish cell line

A pathogenic virus (RGV), isolated from diseased pig frog Rana grylio with lethal syndrome, was investigated with regard to morphogenesis and cellular interactions in EPC cells, a cell Line from fish. Different stages of virus amplification, maturation and assembly were observed at nucleus, cytoplasm and cellular membranes. The matured virus particles, were not only distributed diffusely in nucleus, cytoplasm and cellular surface, but also aggregated as pseudocrystalline arrays in the cytoplasm. Virions were released by budding from the plasma membranes, or following cell lysis. Various types of cell damage, such as small vacuoles, spherical inclusions, and swollen and empty mitochondria, were also found. Some typical characteristics of RGV, such as the symmetrical shape of the virions, replication process involving both nuclear and cytoplasmic phases, budding release from cellular membrane and intracellular membrane, viromatrix and paracrystalline aggregation in cytoplasm, and its acute pathogenic effects, were observed to be similar to that of other iridoviruses. Therefore, the RGV appears to be a member of the Iridoviridae based on these studies. (C) 1999 Elsevier Science B.V. All rights reserved.

An outlook of heavy ion driven plasma research at IMP-Lanzhou

Since the successful completion of the cooling storage ring (CSR) project in China at the end of 2007, high qualitative heavy ion beams with energy ranging from keV to GeV/u have been available at the Heavy Ion Research Facility at Lanzhou (HIRFL). More than 10(9) 1 GeVlu C6+ particles or 10(8) 235 MeV/u Xe particles can be stored in the CSR main-ring and extracted within hundred nano-seconds during the test running, the beam parameters will be improved in the coming years so that high energy density (HED) conditions could be achieved and investigated there. Recent scientific results from the experiments relevant to plasma research on HIRFL are summarized. Dense plasma research with intense heavy ion beams of CSR is proposed here.

Receptor and G betagamma isoform-specific interactions with G protein-coupled receptor kinases.

The G protein-coupled receptor (GPCR) kinases (GRKs) phosphorylate and desensitize agonist-occupied GPCRs. GRK2-mediated receptor phosphorylation is preceded by the agonist-dependent membrane association of this enzyme. Previous in vitro studies with purified proteins have suggested that this translocation may be mediated by the recruitment of GRK2 to the plasma membrane by its interaction with the free betagamma subunits of heterotrimeric G proteins (G betagamma). Here we demonstrate that this mechanism operates in intact cells and that specificity is imparted by the selective interaction of discrete pools of G betagamma with receptors and GRKs. Treatment of Cos-7 cells transiently overexpressing GRK2 with a beta-receptor agonist promotes a 3-fold increase in plasma membrane-associated GRK2. This translocation of GRK2 is inhibited by the carboxyl terminus of GRK2, a known G betagamma sequestrant. Furthermore, in cells overexpressing both GRK2 and G beta1 gamma2, activation of lysophosphatidic acid receptors leads to the rapid and transient formation of a GRK/G betagamma complex. That G betagamma specificity exists at the level of the GPCR and the GRK is indicated by the observation that a GRK2/G betagamma complex is formed after agonist occupancy of the lysophosphatidic acid and beta-adrenergic but not thrombin receptors. In contrast to GRK2, GRK3 forms a G betagamma complex after stimulation of all three GPCRs. This G betagamma binding specificity of the GRKs is also reflected at the level of the purified proteins. Thus the GRK2 carboxyl terminus binds G beta1 and G beta2 but not G beta3, while the GRK3 fusion protein binds all three G beta isoforms. This study provides a direct demonstration of a role for G betagamma in mediating the agonist-stimulated translocation of GRK2 and GRK3 in an intact cellular system and demonstrates isoform specificity in the interaction of these components.

Effect of ritonavir-induced cytochrome P450 3A4 inhibition on plasma fentanyl concentrations during patient-controlled epidural labor analgesia: a pharmacokinetic simulation.

BACKGROUND: Ritonavir inhibition of cytochrome P450 3A4 decreases the elimination clearance of fentanyl by 67%. We used a pharmacokinetic model developed from published data to simulate the effect of sample patient-controlled epidural labor analgesic regimens on plasma fentanyl concentrations in the absence and presence of ritonavir-induced cytochrome P450 3A4 inhibition. METHODS: Fentanyl absorption from the epidural space was modeled using tanks-in-series delay elements. Systemic fentanyl disposition was described using a three-compartment pharmacokinetic model. Parameters for epidural drug absorption were estimated by fitting the model to reported plasma fentanyl concentrations measured after epidural administration. The validity of the model was assessed by comparing predicted plasma concentrations after epidural administration to published data. The effect of ritonavir was modeled as a 67% decrease in fentanyl elimination clearance. Plasma fentanyl concentrations were simulated for six sample patient-controlled epidural labor analgesic regimens over 24 h using ritonavir and control models. Simulated data were analyzed to determine if plasma fentanyl concentrations producing a 50% decrease in minute ventilation (6.1 ng/mL) were achieved. RESULTS: Simulated plasma fentanyl concentrations in the ritonavir group were higher than those in the control group for all sample labor analgesic regimens. Maximum plasma fentanyl concentrations were 1.8 ng/mL and 3.4 ng/mL for the normal and ritonavir simulations, respectively, and did not reach concentrations associated with 50% decrease in minute ventilation. CONCLUSION: Our model predicts that even with maximal clinical dosing regimens of epidural fentanyl over 24 h, ritonavir-induced cytochrome P450 3A4 inhibition is unlikely to produce plasma fentanyl concentrations associated with a decrease in minute ventilation.

Interactions of Lipoprotein(a) with the Plasminogen System: Mechanisms and Pathophysiological Consequences

Elevated plasma concentrations of lipoprotein(a) (Lp(a)) are associated with increased risk of atherothrombotic disease. Lp(a) is a unique lipoprotein consisting of a low density lipoprotein-like moiety covalently linked to apolipoprotein(a) (apo(a)), a homologue of the fibrinolytic proenzyme plasminogen. Apo(a) is extremely heterogeneous in size with small isoforms being independently associated with increased cardiovascular risk. Several in vitro and in vivo studies have shown that Lp(a)/apo(a) can inhibit tissue-type plasminogen activator (tPA)-mediated plasminogen activation on fibrin surfaces, although the mechanism of inhibition by apo(a) remains controversial. Essential to fibrin clot lysis are a number of plasmin-dependent positive feedback reactions that enhance the efficiency of plasminogen activation, including the plasmin-mediated conversion of Glu1-plasminogen to Lys78-plasminogen. Additionally, abnormal fibrin clot structures have been associated with both an increased risk of cardiovascular disease and elevated Lp(a) levels. Similarly, oxidized phospholipids have been implicated in the development of cardiovascular disease, and are not only preferentially carried by Lp(a) in the plasma but have also been shown to covalently-modify both apo(a) and plasminogen. In this thesis, we built upon the understanding of the role of apo(a) in plasminogen activation on the fibrin/degraded fibrin surface by determining that: (i) apo(a) inhibits plasmin-mediated Glu1-plasminogen to Lys78-plasminogen conversion and identifying the critical domains in apo(a) responsible for this effect, (ii) apo(a) isoform size does not affect either the inhibition of tPA-mediated plasminogen activation or the inhibition of plasmin-mediated Glu1-plasminogen to Lys78-plasminogen conversion, (iii) apo(a) modifies fibrin clot structure to form more dense clots with thinner fibers and reduced permeability, modifications that enhance the ability of apo(a) to inhibit tPA-mediated plasminogen activation and (iv) the phosphorus content of apo(a) affects its ability to inhibit tPA-mediated plasminogen activation and the phosphorus content of plasminogen affects its ability to be activated by tPA. By understanding these individual reactions, each of which has the potential to affect the broader fibrin clot lysis process, we have expanded our understanding of the overall effect of Lp(a)/apo(a) in the inhibition of plasminogen activation on the fibrin/degraded fibrin surface and thus broadened our understanding of how Lp(a)/apo(a) may mediate the inhibition of thrombolysis in vivo.

Bright Multi-keV Harmonic Generation from Relativistically Oscillating Plasma Surfaces

The first evidence of x-ray harmonic radiation extending to 3.3 A, 3.8 keV (order n > 3200) from petawatt class laser-solid interactions is presented, exhibiting relativistic limit efficiency scaling (eta similar to n(-2.5)-n(-3)) at multi-keV energies. This scaling holds up to a maximum order, n(RO)similar to 8(1/2)gamma(3), where gamma is the relativistic Lorentz factor, above which the first evidence of an intensity dependent efficiency rollover is observed. The coherent nature of the generated harmonics is demonstrated by the highly directional beamed emission, which for photon energy h nu > 1 keV is found to be into a cone angle similar to 4 degrees, significantly less than that of the incident laser cone (20 degrees).

Plasma ionization through wave-particle interaction in a capacitively coupled radio-frequency discharge

Phase resolved optical emission spectroscopy, with high temporal resolution, shows that wave-particle interactions play a fundamental role in sustaining capacitively coupled rf plasmas. The measurements are in excellent agreement with a simple particle-in-cell simulation. Excitation and ionization mechanisms are dominated by beam-like electrons, energized through the advancing and retreating electric fields of the rf sheath. The associated large-amplitude electron waves, driven by a form of two-stream instability, result in power dissipation through electron trapping and phase mixing. (c) 2007 American Institute of Physics.

Proton Probing Measurement of Electric and Magnetic Fields Generated by ns and ps Laser-Matter Interactions

The use of laser-accelerated protons as a particle probe for the detection of electric fields in plasmas has led in recent years to a wealth of novel information regarding the ultrafast plasma dynamics following high intensity laser-matter interactions. The high spatial quality and short duration of these beams have been essential to this purpose. We will discuss some of the most recent results obtained with this diagnostic at the Rutherford Appleton Laboratory (UK) and at LULI - Ecole Polytechnique (France), also applied to conditions of interest to conventional Inertial Confinement Fusion. In particular, the technique has been used to measure electric fields responsible for proton acceleration from solid targets irradiated with ps pulses, magnetic fields formed by ns pulse irradiation of solid targets, and electric fields associated with the ponderomotive channelling of ps laser pulses in under-dense plasmas.

Plasma Ionization in Low-Pressure Radio-Frequency Discharges. Part I: Optical Measurements

The electron dynamics in the low-pressure operation regime ($«$ 5 Pa) of a neon capacitively coupled plasma is investigated using phase-resolved optical emission spectroscopy. Plasma ionization and sustainment mechanisms are governed by the expanding and contracting sheath and complex wave–particle interactions. Electrons are energized through the advancing and retreating electric field of the RF sheath. The associated interaction of energetic sheath electrons with thermal bulk plasma electrons drives a two-stream instability also dissipating power in the plasma.

Plasma Ionization in Low-Pressure Radio-Frequency Discharges—Part II: Particle-in-Cell Simulation

Plasma ionization in the low-pressure operation regime ( $«$ 5 Pa) of RF capacitively coupled plasmas (CCPs) is governed by a complex interplay of various mechanisms, such as field reversal, sheath expansion, and wave–particle interactions. In a previous paper, it was shown that experimental observations in a hydrogen CCP operated at 13.56 MHz are qualitatively well described in a 1-D symmetrical particle-in-cell (PIC) simulation. In this paper, a spherical asymmetrical PIC simulation that is closer to the conditions of the highly asymmetrical experimental device is used to simulate a low-pressure neon CCP operated at 2 MHz. The results show a similar behavior, with pronounced ionization through field reversal, sheath expansion, and wave–particle interactions, and can be exploited for more accurate quantitative comparisons with experimental observations.

Weakly nonlinear vertical dust grain oscillations in dusty plasma crystals in the presence of a magnetic field

The weakly nonlinear regime of transverse paramagnetic dust grain oscillations in dusty (complex) plasma crystals is discussed. The nonlinearity, which is related to the sheath electric/magnetic field(s) and to the intergrain (electrostatic/magnetic dipole) interactions, is shown to lead to the generation of phase harmonics and, in the case of propagating transverse dust-lattice modes, to the modulational instability of the carrier wave due to self-interaction. The stability profile depends explicitly on the form of the electric and magnetic fields in the plasma sheath. The long term evolution of the modulated wave packet, which is described by a nonlinear Schrodinger-type equation, may lead to propagating localized envelope structures whose exact forms are presented and discussed. Explicit suggestions for experimental investigations are put forward. (C) 2004 American Institute of Physics.