Transient expression of rabies virus G-glycoprotein using BHK-21 cells cultured in suspension

Objective To assess the expression of rabies virus G-glycoprotein (RVGP) expression using Semliki Forest virus as a vector in combination with BHK-21 cells cultured in suspension. Results A multilevel factorial design was used to quantify effects of temperature (33–37 C), fresh medium addition after the viral adsorption step (100–200 % with respect to the initial cell suspension volume before infection) and harvest time (8–40 h) on RVGP production. Experimental runs were performed in 24-well cell culture plates at a multiplicity of infection (MOI) of 16. An additional experiment in spinner-flask was performed at MOI of 9, using the optimal conditions determined in cell culture plates. Values for temperature, fresh medium addition and harvest time of 33 C, 100 % and 16 h, respectively, ensured the optimal RVGP production in culture plates. The volumetric yield (239 ng ml-1 ) in these conditions was higher than that reported previously for adherent cell culture. In spinner-flasks, the volumetric yield was improved (559 ng ml-1 ). Conclusion These results establish the basis for designing bioprocess to produce RVGP.

Multiplex suspension array for human anti-carbohydrate antibody profiling

Glycan-binding antibodies form a significant subpopulation of both natural and acquired antibodies and play an important role in various immune processes. They are for example involved in innate immune responses, cancer, autoimmune diseases, and neurological disorders. In the present study, a microsphere-based flow-cytometric immunoassay (suspension array) was applied for multiplexed detection of glycan-binding antibodies in human serum. Several approaches for immobilization of glycoconjugates onto commercially available fluorescent microspheres were compared, and as the result, the design based on coupling of end-biotinylated glycopolymers has been selected. This method requires only minute amounts of glycans, similar to a printed glycan microarray. The resulting glyco-microspheres were used for detection of IgM and IgG antibodies directed against ABO blood group antigens. The possibility of multiplexing this assay was demonstrated with mixtures of microspheres modified with six different ABO related glycans. Multiplexed detection of anti-glycan IgM and IgG correlated well with singleplex assays (Pearson's correlation coefficient r = 0.95-0.99 for sera of different blood groups). The suspension array in singleplex format for A/B trisaccharide, H(di) and Le(x) microspheres corresponded well to the standard ELISA (r > 0.94). Therefore, the described method is promising for rapid, sensitive, and reproducible detection of anti-glycan antibodies in a multiplexed format.

Design, Simulation und Erprobung einer aktiven Sitzaufhängung für Nutzfahrzeuge

Nutzfahrzeuge müssen oft durch sehr unebenes Gelände gefahren werden. In diesem Fall wird der Fahrer starken Vibrationen ausgesetzt, die von der Fahrzeugkarosserie durch die Sitzaufhängung auf ihn wirken. Um diese Schwingungen zu verringern, werden die Sitzaufhängungen in der Regel mit Feder-Dämpfer-Systemen ausgerüstet. Jedoch erreichen die passiven Systeme vor allem bei niederfrequenten Schwingungen ihre physikalischen Grenzen. Eine wesentliche Verbesserung des Sitzkomforts kann unter solchen Anregungsbedingungen nur mit einer aktiven Sitzaufhängung erreicht werden. In diesem Beitrag wird ein neuartiges aktives System für die Sitzaufhängung auf Basis von elektrorheologischen Flüssigkeiten vorgestellt. Außerdem werden die theoretischen Grundlagen für die Modellierung der beschriebenen aktiven Sitzaufhängung dargestellt. Anschließend werden die Simulationsergebnisse mit den Messergebnissen unter realen Betriebsbedingungen verglichen. Die Repräsentation der Ergebnisse mit Hilfe der im Bereich der Sitztechnik weit verbreiteten SEAT-Werten (Seat Effective Amplitude Transmissibility) zeigt das Potenzial des entwickelten Systems zur aktiven Reduktion der Schwingungsbelastung des Fahrers und ermöglicht seine objektive Bewertung.

Design and implementation of a modular interface to integrate CLP and tabled execution

Logic programming (LP) is a family of high-level programming languages which provides high expressive power. With LP, the programmer writes the properties of the result and / or executable specifications instead of detailed computation steps. Logic programming systems which feature tabled execution and constraint logic programming have been shown to increase the declarativeness and efficiency of Prolog, while at the same time making it possible to write very expressive programs. Tabled execution avoids infinite failure in some cases, while improving efficiency in programs which repeat computations. CLP reduces the search tree and brings the power of solving (in)equations over arbitrary domains. Similarly to the LP case, CLP systems can also benefit from the power of tabling. Previous implementations which take ful advantage of the ideas behind tabling (e.g., forcing suspension, answer subsumption, etc. wherever it is necessary to avoid recomputation and terminate whenever possible) did not offer a simple, well-documented, easy-to-understand interface. This would be necessary to make the integratation of arbitrary CLP solvers into existing tabling systems possible. This clearly hinders a more widespread usage of the combination of both facilities. In this thesis we examine the requirements that a constraint solver must fulfill in order to be interfaced with a tabling system. We propose and implement a framework, which we have called Mod TCLP, with a minimal set of operations (e.g., entailment checking and projection) which the constraint solver has to provide to the tabling engine. We validate the design of Mod TCLP by a series of use cases: we re-engineer a previously existing tabled constrain domain (difference constraints) which was connected in an ad-hoc manner with the tabling engine in Ciao Prolog; we integrateHolzbauer’s CLP(Q) implementationwith Ciao Prolog’s tabling engine; and we implement a constraint solver over (finite) lattices. We evaluate its performance with several benchmarks that implement a simple abstract interpreter whose fixpoint is reached by means of tabled execution, and whose domain operations are handled by the constraint over (finite) lattices, where TCLP avoids recomputing subsumed abstractions.---ABSTRACT---La programación lógica con restricciones (CLP) y la tabulación son extensiones de la programación lógica que incrementan la declaratividad y eficiencia de Prolog, al mismo tiempo que hacen posible escribir programasmás expresivos. Las implementaciones anteriores que integran completamente ambas extensiones, incluyendo la suspensión de la ejecución de objetivos siempre que sea necesario, la implementación de inclusión (subsumption) de respuestas, etc., en todos los puntos en los que sea necesario para evitar recomputaciones y garantizar la terminación cuando sea posible, no han proporcionan una interfaz simple, bien documentada y fácil de entender. Esta interfaz es necesaria para permitir integrar resolutores de CLP arbitrarios en el sistema de tabulación. Esto claramente dificulta un uso más generalizado de la integración de ambas extensiones. En esta tesis examinamos los requisitos que un resolutor de restricciones debe cumplir para ser integrado con un sistema de tabulación. Proponemos un esquema (y su implementación), que hemos llamadoMod TCLP, que requiere un reducido conjunto de operaciones (en particular, y entre otras, entailment y proyección de almacenes de restricciones) que el resolutor de restricciones debe ofrecer al sistema de tabulación. Hemos validado el diseño de Mod TCLP con una serie de casos de uso: la refactorización de un sistema de restricciones (difference constraints) previamente conectado de un modo ad-hoc con la tabulación de Ciao Prolog; la integración del sistema de restricciones CLP(Q) de Holzbauer; y la implementación de un resolutor de restricciones sobre retículos finitos. Hemos evaluado su rendimiento con varios programas de prueba, incluyendo la implementación de un intérprete abstracto que alcanza su punto fijo mediante el sistema de tabulación y en el que las operaciones en el dominio son realizadas por el resolutor de restricciones sobre retículos (finitos) donde TCLP evita la recomputación de valores abstractos de las variables ya contenidos en llamadas anteriores.

Modelling of entrainment in industrial flotation cells: the effect of solids suspension

Solids concentration and particle size distribution gradually change in the vertical dimension of industrial flotation cells, subject primarily to the flotation cell size and design and the cell operating conditions. As entrainment is a two-step process and involves only the suspended solids in the top pulp region near the pulp-froth interface, the solids suspension characteristics have a significant impact on the overall entrainment. In this paper, a classification function is proposed to describe the state of solids suspension in flotation cells, similar to the definition of degree of entrainment for classification in the froth phase found in the literature. A mathematical model for solids suspension is also developed, in which the classification function is expressed as an exponential function of the particle size. Experimental data collected from three different Outokumpu tank flotation cells in three different concentrators are well fitted by the proposed exponential model. Under the prevailing experimental conditions, it was found that the solids content in the top region was relatively independent of cell operating conditions such as froth height and air rate but dependent on the cell size. Moreover, the results obtained from the total solids tend to be similar to those from a particular gangue mineral and hence may be applied to all minerals in entrainment calculation. (C) 2004 Elsevier Ltd. All rights reserved.

The development of a novel Bio-MEMS filtration chip for the separation of specific cells in fluid suspension

In the clinical/microbiological laboratory there are currently several ways of separating specific cells from a fluid suspension. Conventionally cells can be separated based on size, density, electrical charge, light-scattering properties, and antigenic surface properties. Separating cells using these parameters can require complex technologies and specialist equipment. This paper proposes new Bio-MEMS (microelectromechanical systems) filtration chips manufactured using deep reactive ion etching (DRIE) technology that, when used in conjunction with an optical microscope and a syringe, can filter and grade cells for size without the requirement for additional expensive equipment. These chips also offer great versatility in terms of design and their low cost allows them to be disposable, eliminating sample contamination. The pumping mechanism, unlike many other current filtration techniques, leaves samples mechanically and chemically undamaged. In this paper the principles behind harnessing passive pumping are explored, modelled, and validated against empirical data, and their integration into a microfluidic device to separate cells from a mixed population suspension is described. The design, means of manufacture, and results from preliminary tests are also presented. © IMechE 2007.

Fopid Controller Design for Robust Performance Using Particle Swarm Optimization

Mathematics Subject Classification: 26A33; 93C15, 93C55, 93B36, 93B35, 93B51; 03B42; 70Q05; 49N05

Uniform design for the optimization of Al2O3 nanofilms produced by electrophoretic deposition

Surface modification by means of nanostructures is of interest to enhance boiling heat transfer in various applications including the organic Rankine cycle (ORC). With the goal of obtaining rough and dense aluminum oxide (Al2O3) nanofilms, the optimal combination of process parameters for electrophoretic deposition (EPD) based on the uniform design (UD) method is explored in this paper. The detailed procedures for the EPD process and UD method are presented. Four main influencing conditions controlling the EPD process were identified as nanofluid concentration, deposition time, applied voltage and suspension pH. A series of tests were carried out based on the UD experimental design. A regression model and statistical analysis were applied to the results. Sensitivity analyses of the effect of the four main parameters on the roughness and deposited mass of Al2O3 films were also carried out. The results showed that Al2O3 nanofilms were deposited compactly and uniformly on the substrate. Within the range of the experiments, the preferred combination of process parameters was determined to be nanofluid concentration of 2 wt.%, deposition time of 15 min, applied voltage of 23 V and suspension pH of 3, yielding roughness and deposited mass of 520.9 nm and 161.6 × 10− 4 g/cm2, respectively. A verification experiment was carried out at these conditions and gave values of roughness and deposited mass within 8% error of the expected ones as determined from the UD approach. It is concluded that uniform design is useful for the optimization of electrophoretic deposition requiring only 7 tests compared to 49 using the orthogonal design method.

Design construction and testing of a heat loop for study of fouling and its interrelationship to corrosion in heat exchanger tubes

A heat loop suitable for the study of thermal fouling and its relationship to corrosion processes was designed, constructed and tested. The design adopted was an improvement over those used by such investigators as Hopkins and the Heat Transfer Research Institute in that very low levels of fouling could be detected accurately, the heat transfer surface could be readily removed for examination and the chemistry of the environment could be carefully monitored and controlled. In addition, an indirect method of electrical heating of the heat transfer surface was employed to eliminate magnetic and electric effects which result when direct resistance heating is employed to a test section. The testing of the loop was done using a 316 stainless steel test section and a suspension of ferric oxide and water in an attempt to duplicate the results obtained by Hopkins. Two types of thermal ·fouling resistance versus time curves were obtained . (i) Asymptotic type fouling curve, similar to the fouling behaviour described by Kern and Seaton and other investigators, was the most frequent type of fouling curve obtained. Thermal fouling occurred at a steadily decreasing rate before reaching a final asymptotic value. (ii) If an asymptotically fouled tube was cooled with rapid cir- ·culation for periods up to eight hours at zero heat flux, and heating restarted, fouling recommenced at a high linear rate. The fouling results obtained were observed to be similar and 1n agreement with the fouling behaviour reported previously by Hopkins and it was possible to duplicate quite closely the previous results . This supports the contention of Hopkins that the fouling results obtained were due to a crevice corrosion process and not an artifact of that heat loop which might have caused electrical and magnetic effects influencing the fouling. The effects of Reynolds number and heat flux on the asymptotic fouling resistance have been determined. A single experiment to study the effect of oxygen concentration has been carried out. The ferric oxide concentration for most of the fouling trials was standardized at 2400 ppM and the range of Reynolds number and heat flux for the study was 11000-29500 and 89-121 KW/M², respectively.

New design of electrostatic mirror actuators for application in high-precision interferometry

We describe a new geometry for electrostatic actuators to be used in sensitive laser interferometers, suited for prototype and table top experiments related to gravitational wave detection with mirrors of 100 g or less. The arrangement consists of two plates at the sides of the mirror (test mass), and therefore does not reduce its clear aperture as a conventional electrostatic drive (ESD) would do. Using the sample case of the AEI-10 m prototype interferometer, we investigate the actuation range and the influence of the relative misalignment of the ESD plates with respect to the test mass. We find that in the case of the AEI-10 m prototype interferometer, this new kind of ESD could provide a range of 0.28 mu m when operated at a voltage of 1 kV. In addition, the geometry presented is shown to provide a reduction factor of about 100 in the magnitude of the actuator motion coupling to the test mass displacement. We show that therefore in the specific case of the AEI-10 m interferometer, it is possible to mount the ESD actuators directly on the optical table without spoiling the seismic isolation performance of the triple stage suspension of the main test masses.