Regulation of neuronal excitability through pumilio-dependent control of a sodium channel gene

Dynamic changes in synaptic connectivity and strength, which occur during both embryonic development and learning, have the tendency to destabilize neural circuits. To overcome this, neurons have developed a diversity of homeostatic mechanisms to maintain firing within physiologically defined limits. In this study, we show that activity-dependent control of mRNA for a specific voltage-gated Na+ channel [encoded by paralytic (para)] contributes to the regulation of membrane excitability in Drosophila motoneurons. Quantification of para mRNA, by real-time reverse-transcription PCR, shows that levels are significantly decreased in CNSs in which synaptic excitation is elevated, whereas, conversely, they are significantly increased when synaptic vesicle release is blocked. Quantification of mRNA encoding the translational repressor pumilio (pum) reveals a reciprocal regulation to that seen for para. Pumilio is sufficient to influence para mRNA. Thus, para mRNA is significantly elevated in a loss-of-function allele of pum (pumbemused), whereas expression of a full-length pum transgene is sufficient to reduce para mRNA. In the absence of pum, increased synaptic excitation fails to reduce para mRNA, showing that Pum is also necessary for activity-dependent regulation of para mRNA. Analysis of voltage-gated Na+ current (INa) mediated by para in two identified motoneurons (termed aCC and RP2) reveals that removal of pum is sufficient to increase one of two separable INa components (persistent INa), whereas overexpression of a pum transgene is sufficient to suppress both components (transient and persistent). We show, through use of anemone toxin (ATX II), that alteration in persistent INa is sufficient to regulate membrane excitability in these two motoneurons

Information retrieval system

Generalized information storage and retrieval system capable of generating and maintaining a file, gathering statistics, sorting output, and generating final reports for output is reviewed. File generation and file maintenance programs written for the system are general purpose routines

Hepatoma cell density promotes claudin-1 and scavenger receptor BI expression and hepatitis C virus internalization.

Hepatitis C virus (HCV) entry occurs via a pH- and clathrin-dependent endocytic pathway and requires a number of cellular factors, including CD81, the tight-junction proteins claudin 1 (CLDN1) and occludin, and scavenger receptor class B member I (SR-BI). HCV tropism is restricted to the liver, where hepatocytes are tightly packed. Here, we demonstrate that SR-BI and CLDN1 expression is modulated in confluent human hepatoma cells, with both receptors being enriched at cell-cell junctions. Cellular contact increased HCV pseudoparticle (HCVpp) and HCV particle (HCVcc) infection and accelerated the internalization of cell-bound HCVcc, suggesting that the cell contact modulation of receptor levels may facilitate the assembly of receptor complexes required for virus internalization. CLDN1 overexpression in subconfluent cells was unable to recapitulate this effect, whereas increased SR-BI expression enhanced HCVpp entry and HCVcc internalization, demonstrating a rate-limiting role for SR-BI in HCV internalization

Supersonic combustion ramjet propulsion experiments in a shock tunnel

Measurements have been made of the propulsive effect of supersonic combustion ramjets incorporated into a simple axisymmetric model in a free piston shock tunnel. The nominal Mach number was 6, and the stagnation enthalpy varied from 2.8 MJ kg(exp -1) to 8.5 MJ kg(exp -1). A mixture of 13 percent silane and 87 percent hydrogen was used as fuel, and experiments were conducted at equivalence ratios up to approximately 0.8. The measurements involved the axial force on the model, and were made using a stress wave force balance, which is a recently developed technique for measuring forces in shock tunnels. A net thrust was experienced up to a stagnation enthalpy of 3.7 MJ kg(exp -1), but as the stagnation enthalpy increased, an increasing net drag was recorded. pitot and static pressure measurements showed that the combustion was supersonic. The results were found to compare satisfactorily with predictions based on established theoretical models, used with some simplifying approximations. The rapid reduction of net thrust with increasing stagnation enthalpy was seen to arise from increasing precombustion temperature, showing the need to control this variable if thrust performance was to be maintained over a range of stagnation enthalpies. Both the inviscid and viscous drag were seen to be relatively insensitive to stagnation enthalpy, with the combustion chambers making a particularly significant contribution to drag. The maximum fuel specific impulse achieved in the experiments was only 175 sec., but the theory indicates that there is considerable scope for improvement on this through aerodynamic design

Simulating field-aligned diffusion of a cosmic ray gas

The macroscopic behaviour of cosmic rays in turbulent magnetic fields is discussed. An implementation of anisotropic diffusion of cosmic rays with respect to the magnetic field in a non-conservative, high-order, finite-difference magnetohydrodynamic code is discussed. It is shown that the standard implementation fails near singular X-points of the magnetic field, which are common if the field is random. A modification to the diffusion model for cosmic rays is described and the resulting telegraph equation (implemented by solving a dynamic equation for the diffusive flux of cosmic rays) is used; it is argued that this modification may better describe the physics of cosmic ray diffusion. The present model reproduces several processes important for the propagation and local confinement of cosmic rays, including spreading perpendicular to the local large-scale magnetic field, controlled by the random-to-total magnetic field ratio, and the balance between cosmic ray pressure and magnetic tension. Cosmic ray diffusion is discussed in the context of a random magnetic field produced by turbulent dynamo action. It is argued that energy equipartition between cosmic rays and other constituents of the interstellar medium do not necessarily imply that cosmic rays play a significant role in the balance of forces.Comment: 11 pages, 12 figures, a major revision; accepted for publication in MNRA

Numerical Simulation of a Ludwieg-tube Fuel Delivery System for Scramjet Experiments in Shock Tunnels

The T4 shock tunnel at The University of Queensland is regularly used to perform supersonic combustion experiments. The fuel for the test model is supplied using a Ludwieg-tube delivery system. A combination of theoretical modelling and calibration tests is used to determine the mass flow-rate of fuel for given Ludwieg tube initials conditions and the measured pressure in the plenum chamber that supplies the fuel to the model. The theoretical model used in the calibration procedure is presented. The goal of this project is to check the suitability of the modelling assumptions by simulating the complete Ludwieg-tube system using the one-dimensional Lagrangian computer code, L1d. Simulation of the fuel delivery system is done separately for viscous and inviscid flow with conditions replicating those used in supersonic combustion tests in T4. The steady plenum pressures from the simulations of inviscid and viscous flow are, on average, within ±7% and ±4%, respectively, of the values measured experimentally. Further, the fuel mass flow-rates obtained from viscous simulations are, on average, within ±13% of the experimental values

Polarization restricts hepatitis C virus entry into HepG2 hepatoma cells

The primary reservoir for hepatitis C virus (HCV) replication is believed to be hepatocytes, which are highly polarized with tight junctions (TJ) separating their basolateral and apical domains. HepG2 cells develop polarity over time, resulting in the formation and remodeling of bile canalicular (BC) structures. HepG2 cells expressing CD81 provide a model system to study the effects of hepatic polarity on HCV infection. We found an inverse association between HepG2-CD81 polarization and HCV pseudoparticle entry. As HepG2 cells polarize, discrete pools of claudin-1 (CLDN1) at the TJ and basal/lateral membranes develop, consistent with the pattern of receptor staining observed in liver tissue. The TJ and nonjunctional pools of CLDN1 show an altered association with CD81 and localization in response to the PKA antagonist Rp-8-Br-cyclic AMPs (cAMPs). Rp-8-Br-cAMPs reduced CLDN1 expression at the basal membrane and inhibited HCV infection, supporting a model where the nonjunctional pools of CLDN1 have a role in HCV entry. Treatment of HepG2 cells with proinflammatory cytokines, tumor necrosis factor alpha and gamma interferon, perturbed TJ integrity but had minimal effect(s) on cellular polarity and HCV infection, suggesting that TJ integrity does not limit HCV entry into polarized HepG2 cells. In contrast, activation of PKC with phorbol ester reduced TJ integrity, ablated HepG2 polarity, and stimulated HCV entry. Overall, these data show that complex hepatocyte-like polarity alters CLDN1 localization and limits HCV entry, suggesting that agents which disrupt hepatocyte polarity may promote HCV infection and transmission within the liver

Visualization Of Supersonic Flows In Shock Tunnels, Using The Background Oriented Schlieren Technique

Visualisation of supersonic compressible flows using the Background Oriented Schlieren (BOS) technique is presented. Results from experiments carried out in a reflected shock tunnel with models of a 20-degree semi-vertex angle circular cone and a re-entry body in the test section are presented. This technique uses a simple optical set-up consisting of a structured background pattern, an electronic camera with a high shutter speed and a high intensity light source. Tests were conducted with a Mach 4 conical nozzle, with nozzle supply pressure of 2 MPa and nozzle supply temperature of 2000 K respectively. The images captured during the test were compared using PIV style image processing code. The intensity of light at each point in the processed image was proportional to the density at that point. Qualitative visualization of shock shapes, with images clearly indicating regions of subsonic and supersonic flows was achieved. For the cone, the shock angle measured from the BOS image agreed with theoretical calculations to within 0.5 degrees. Shock standoff distances could be measured from the BOS image for the re-entry body

Scramjets and Shock Tunnels: The Queensland Experience

This article reports on the use of a shock tunnel to study the operation of scramjet powered configurations at sub-orbital velocities above 2 km/s. Thrust, as given by a net thrust equation, is used as a figure of merit throughout the study. After a short description of the shock tunnel used and its operating characteristics, experiments on the combustion release of heat in a constant area duct with hydrogen fuel are reviewed. The interaction between heat release in the combustion wake and the walls of the duct produced pressure distributions which followed a binary scaling law, and indicated that the theoretically expected heat release could be realized in practice, albeit with high pressure or long combustion ducts. This heat release, combined with attainable thrust nozzle characteristics and a modest level of configuration drag, indicated that positive thrust levels could be obtained well into the sub-orbital range of velocities. Development of a stress wave force balance for use in shock tunnels allowed the net thrust generated to be measured for integrated scramjet configurations and, although the combination of model size and shock tunnel operating pressure prevented complete combustion of hydrogen, the cruise condition of zero net thrust was achieved at 2.5 km/s with one configuration, while net thrust was produced with another configuration using an ignition promoter in hydrogen fuel. Nevertheless, the combination of boundary layer separation induced inlet choking and limited operating pressure levels prevented realization of the thrust potential of the fuel. This problem may be alleviated by recent increases in the shock tunnel operating pressures, and by promising research involving inlet injection of the fuel. Research on the drag component of the net thrust equation resulted from the development of a fast response skin friction gauge. It was found that existing theories of turbulent boundary skin friction predicted the skin friction when combustion of hydrogen occurred outside the boundary layer, but combustion within the boundary layer dramatically reduced the skin friction. Finally, for the first time in the world, supersonic combustion was produced in a free flight experiment. This experiment validated shock tunnel results at stagnation enthalpies near 3 MJ/k