V antigen-polyhistidine fusion peptide: binding to LcrH and active immunity against plague

The structural gene for V antigen (lcrV) is known to be encoded within the lcrGVH-yopBD operon of the approximately 70-kb low-calcium-response or Lcr plasmid of Yersinia pestis. This 37-kDa monomeric peptide was reported to provide active immunity in mice, suppress inflammatory cytokines, and regulate expression of the low calcium response (Lcr+). Here we describe pVHB62, encoding a polyhistidine-V antigen fusion peptide (Vh) and linked LcrH. Vh underwent degradation from both the C terminus and N terminus during classical chromatographic fractionation but remained intact within two compartments during Ni2+ affinity chromatography. The first was homogeneous, capable of active immunization (mouse intravenous 50% lethal dose, > 10(7) bacteria), and stable at 4 degrees C. The second remained bound to the affinity column but could be eluted as a mixture of Vh, LcrH, and low-molecular-weight material by application of 6 M guanidine HCl. This mixture was dialyzed, denatured in 8 M urea, and again applied to the affinity column, which then hound Vh but not LcrH. The latter was recovered and renatured, and low-molecular-weight material was removed by biochemical fractionation. The resulting homogeneous LcrH bound protein AN antigen fusion peptide but not protein A in a sandwich enzyme-linked immunosorbent assay, and this reaction was inhibited by Vh. These observations indicate that LcrH normally binds V antigen in bacterial cytoplasm and suggest that only free LcrH down-regulates expression of the low calcium response.     

A video conference network management system

Setting up either one or multiple conferences and managing the various devices can be a very difficult task. In this article a newly developed distributed video conference management system is explained and tested. © 1998 John Wiley & Sons, Ltd.     

Influence of vanadium concentration on the microstructure and magnetic properties of V-doped ZnO thin films

Vanadium doped ZnO thin films (Zn_1 − xV_xO, where x = 0.05 or x = 0.13) were grown on c-cut sapphire substrates using pulsed laser deposition technique. Their structure and magnetic properties were examined in relation to the doping concentration. All deposited films were highly oriented along the c-axis and exhibited ferromagnetic behavior with a Curie temperature up to 300 K. The crystal structure was found to be better for layers with lower vanadium concentration. The films had a porous fine-grained microstructure and a column-like character as the V concentration was reduced. A weak dependence of magnetization on temperature was observed. The saturation magnetization was found to be strongly dependent on the crystal structure, grain size and V-ion concentration.     

Resistance to lipopolysaccharide mediated by the Yersinia pestis V antigen-polyhistidine fusion peptide: amplification of interleukin-10

We previously showed that injection of homogenous staphylococcal protein A-V antigen fusion peptide into mice delayed allograft rejection and suppressed the major proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and gamma interferon (IFN-gamma) associated with generation of protective granulomas. This study was undertaken to determine if V antigen could prevent endotoxic shock, known to be mediated by excessive production of certain proinflammatory cytokines. After treatment with 50 microg of homogeneous V antigen-polyhistidine fusion peptide (Vh), the 50% lethal dose of purified lipopolysaccharide (LPS) in BALB/c mice immediately rose from 63 microg (normal controls) to 318 microg, fell to near baseline (71 microg) in 6 h, and then slowly rose to a maximum of 566 microg at 48 h before again returning to normal. Injected Vh alone (50 microg) promptly induced the anti-inflammatory cytokine interleukin-10 (IL-10) as well as modest levels of TNF-alpha (an inducer of IL-10) in spleen. Concomitant injection of Vh and an otherwise lethal dose of LPS (200 microg) dramatically decreased levels of TNF-alpha and IFN-gamma in the spleen and peritoneal lavage fluid as compared to values determined for LPS alone. These results would be expected if V antigen directly up-regulated IL-10 that is reported to generally down-regulate proinflammatory cytokines. Mice receiving 200 microg of LPS 48 h after injection of Vh exhibited patterns of cytokine synthesis similar to those observed in endotoxin-tolerant mice, a condition also reported to be mediated by IL-10. These findings suggest that V antigen serves as a virulence factor by amplifying IL-10, thereby repressing proinflammatory cytokines required for expression of cell-mediated immunity.     

Comparison of Air Temperature Calibrations

European national metrology institutes use calibration systems of various types for calibrating thermometers in air. These were compared to each other for the first time in a project organized by the European Association of National Metrology Institutes (EURAMET). This EURAMET P1061 comparison project had two main objectives: (1) to study the equivalence of calibrations performed by different laboratories and (2) to investigate correlations between calibration methods and achievable uncertainties. The comparison was realized using a pair of 100  \(\Omega \) platinum resistance thermometer probes connected to a digital thermometer bridge as the transfer standard. The probes had different dimensions and surface properties. The measurements covered the temperature range between \(-40\,^{\circ }\mathrm{{C}}\) and \(+150\,^{\circ }\mathrm{{C}}\) , but each laboratory chose a subrange most relevant to its scope and performed measurements at five nominal temperature points covering the subrange. To enable comparison between the laboratories, comparison reference functions were determined using weighted least-squares fitting. Various effects related to variations in heat transfer conditions were demonstrated but clear correlations to specific characteristics of calibration system were not identified. Calibrations in air and liquid agreed typically within \(\pm 0.05\,^{\circ }\mathrm{{C}}\) at \(+10\,^{\circ }\mathrm{{C}}\) and \(+80\,^{\circ }\mathrm{{C}}\) . Expanded uncertainties determined by the participants ranged from \(0.02\,^{\circ }\mathrm{{C}}\) to \(0.4\,^{\circ }\mathrm{{C}}\) and they were shown to be realistic in most cases.     

An Effective High-Order Interval Method for Validating Existence and Uniqueness of the Solution of an IVP for an ODE

Validated methods for initial value problems for ordinary differential equations produce bounds that are guaranteed to contain the true solution of a problem. When computing such bounds, these methods verify that a unique solution to the problem exists in the interval of integration and compute a priori bounds for the solution in this interval. A major difficulty in this verification phase is how to take as large a stepsize as possible, subject to some tolerance requirement. We propose a high-order enclosure method for proving existence and uniqueness of the solution and computing a priori bounds.     

An Interval Hermite-Obreschkoff Method for Computing Rigorous Bounds on the Solution of an Initial Value Problem for an Ordinary Differential Equation

To date, the only effective approach for computing guaranteed bounds on the solution of an initial value problem (IVP) for an ordinary differential equation (ODE) has been interval methods based on Taylor series. This paper derives a new approach, an interval Hermite-Obreschkoff (IHO) method, for computing such enclosures. Compared to interval Taylor series (ITS) methods, for the same stepsize and order, our IHO scheme has a smaller truncation error, better stability, and requires fewer Taylor coefficients and high-order Jacobians.The stability properties of the ITS and IHO methods are investigated. We show as an important by-product of this analysis that the stability of an interval method is determined not only by the stability function of the underlying formula, as in a standard method for an IVP for an ODE, but also by the associated formula for the truncation error.