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Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN′ from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains.
The optical study carried out on insulating polymers namely polyethyleneterephthalate (PET) and polyvinylchloride (PVC) has been described. The polymers are exposed to different radiation doses by exposing them to swift heavy ions of carbon (90 MeV), silicon (120 MeV) and nickel (100 MeV) which influence on their optical properties. The studies show that amongst the investigated polymers, PVC and PET have potential for application as dosimeter beyond a threshold dose which is strongly dependent on the nature of the material and the radiation type. The optical micrographs show a distinct change in colour of the sample with increase in radiation dose.
In order to allow an autonomous robot to perform non-trivial tasks like to explore a foreign planet the robot has to have deliberative capabilities like reasoning or planning. Logic-based approaches like the programming and planing language Golog and it successors has been successfully used for such decision-making problems. A drawback of this particular programing language is that their interpreter usually are written in Prolog and run on a Prolog back-end. Such back-ends are usually not available or feasible on resource-limited robot systems. In this paper we present our ideas and first results of a re-implementation of the interpreter based on the Lua scripting language which is available on a wide range of systems including small embedded systems.
This paper aims to evaluate the formability of tailor welded blanks of dual phase (DP600)/transformation induced plasticity (TRIP700) steel sheets. In this work, bead on plate butt joints of 2·5 mm DP600 and 1·2 mm TRIP700 steel sheets were performed using CO2 laser beam welding. Microhardness measurements and transverse tensile testing were carried out to characterise the welds. The formability of base metals and welds were investigated by standard Erichsen test. In a perpendicular tensile test to the weld line, all specimens were fractured at the TRIP base metal, and the strengths were somewhat higher than those of base metal. There was a significant reduction in formability caused by welding of the DP600/TRIP700 steel sheets, and the formability increased with increasing welding speed.
Numerical solution of the heat equation with non-linear, time derivative-dependent source term
(2010)
The mathematical modeling of heat conduction with adsorption effects in coated metal structures yields the heat equation with piecewise smooth coefficients and a new kind of source term. This term is special, because it is non-linear and furthermore depends on a time derivative. In our approach we reformulated this as a new problem for the usual heat equation, without source term but with a new non-linear coefficient. We gave an existence and uniqueness proof for the weak solution of the reformulated problem. To obtain a numerical solution, we developed a semi-implicit and a fully implicit finite volume method. We compared these two methods theoretically as well as numerically. Finally, as practical application, we simulated the heat conduction in coated aluminum fibers with adsorption in the zeolite coating. Copyright © 2010 John Wiley & Sons, Ltd.
Aims: Thrombotic complications due to activation of platelets and plasmatic clotting factors belong still to the most investigated topics in the field of study of patho-physiological mechanisms. Mathematical modeling of thrombotic reactions is established and validated in test cases. Aim of this study is to experimentally evaluate and computationally simulate platelets under the influence of well-defined shear flow conditions. Platelet behaviour and reactions are experimentally reproduced, measured and used for validation of the numerical simulation. Methods: A mathematical model of platelet activation, adhesion and aggregation has been implemented into a finite element CFD (Computational Fluid Dynamics) code. The approach is based on the advective and diffusive transport equations for resting platelets, activated platelets and platelet released agonists. Adhesion rates for the reactive surfaces depend on the hemocompatibility properties of the surface and the local shear rate. Experiments with citrate-anticoagulated freshly-drawn whole blood are performed in a perfusion flow chamber as well as in a system of rotating cylinders for Couette and Taylor-vortex flow. Different biomaterials are used. The activation, drop of platelet concentration, adhesion and aggregation are quantified using scanning electron microscopy (SEM) and flow cytometry. Results: Regions and flow conditions with a high potential for thrombus growth could be identified. The experiments clearly show the influence of the blood contacting material and flow properties. By means of SEM diverse platelet adhesion patterns are observed. Numerical analysis can explain the patterns and the degree of thrombus formation. Conclusion: The numerical method shows good agreement with experimental data indicating a possible prediction of initiation of activation and detection of the local adhesion areas in connection with the role of Von-Willebrand-Factor.
Recently, we introduced and mathematically analysed a new method for grid deformation (Grajewski et al., 2009) [15] we call basic deformation method (BDM) here. It generalises the method proposed by Liao et al. (Bochev et al., 1996; Cai et al., 2004; Liao and Anderson, 1992) [4], [6], [20]. In this article, we employ the BDM as core of a new multilevel deformation method (MDM) which leads to vast improvements regarding robustness, accuracy and speed. We achieve this by splitting up the deformation process in a sequence of easier subproblems and by exploiting grid hierarchy. Being of optimal asymptotic complexity, we experience speed-ups up to a factor of 15 in our test cases compared to the BDM. This gives our MDM the potential for tackling large grids and time-dependent problems, where possibly the grid must be dynamically deformed once per time step according to the user's needs. Moreover, we elaborate on implementational aspects, in particular efficient grid searching, which is a key ingredient of the BDM.
Chemical imaging systems allow the visualisation of the distribution of chemical species on the sensor surface. This work represents a new flexible approach of read out in a light-addressable potentiometric sensor (LAPS) with the help of a digital light processing (DLP) set-up. The DLP, known well for video projectors, consists of a mirror-array MEMS device which allows fast and flexible generation of light patterns. With the help of these light patterns the sensor surface of the LAPS device can be read out sequentially in a raster like scheme (scanning LAPS). The DLP approach has several advantages compared to conventional scanning LAPS set-ups, e.g., the spot size, the shape and the intensity of the light pointer can be changed easily and no mechanical movement is necessary, which reduces the size of the set-up and increases the stability and speed of measurement.
Normative Regulations
(2010)
The Newtonian regime of a recent nonlocal extension of general relativity is investigated. Nonlocality is introduced via a scalar “constitutive” kernel in a special case of the translational gauge theory of gravitation, namely, the teleparallel equivalent of general relativity. In this theory, the nonlocal aspect of gravity simulates dark matter. A nonlocal and nonlinear generalization of Poisson’s equation of Newtonian gravitation is presented. The implications of nonlocality for the gravitational physics in the solar system are briefly studied.
The MicroMed DeBakey ventricular assist device is an axial flow pump designed for providing long-term support to end-stage heartfailure patients. Previously, we presented computational analysis of the blood pump flow. From the analysis, we were able to identify regions of high shear and recirculating flow that may cause blood damage, for example, deformation and fragmentation of the red blood cell (RBC). This mechanical hemolysis can be predicted using a tensor-based blood damage model that is based on the physical properties of the RBCs, for example, the relaxation time of the RBC membrane. However, an extensive and detailed analysis was complicated by the fact that the previous method predicts hemolysis along a finite number of pathlines traversed by the RBCs, possibly omitting parts of the flow domain. Furthermore, it is computationally expensive and is not easily parallelizable.
Here, we propose a new method to estimate hemolysis. The method is based on treating the shape of droplet (tensor) as a field variable, like velocity in the Navier-Stokes system. The governing equation for the RBC shape is treated by least-squares finite element method and the volume conservation of the RBC is augmented by Lagrangian multiplier. Unlike the previous method, the proposed method can visualize areas of high RBC strain that is potentially dangerous for mechanical hemolysis. Also, the amount of plasma-free hemoglobin and, consequently, normalized index of hemolysis can be computed as a byproduct. The method is tested in a simple shear flow for validation and an artery graft flow is chosen to show its potential usefulness. Finally, the method is applied to the blood damage estimation for the pump.
Purpose of Study: Thrombosis-related complications are among the leading causes for morbidity and mortality in patients who depend on artificial organs. For the prediction of platelet behavior both the flow conditions inside the device and the thrombogenic properties of the blood-contacting surfaces must be considered. Platelet reactions under the influence of well-defined shear rates are experimentally evaluated and numerically simulated. The approach is intended for the analysis of VAD and oxygenator design.
Methods Used: A mathematical model of platelet activation, adhesion and aggregation has been implemented into a finite element CFD (Computational Fluid Dynamics) code. The approach is based on the advective and diffusive transport equations for resting and activated platelets and platelet released agonists. Experiments with citrate-anticoagulated freshly-drawn whole blood are performed in a perfusion flow chamber as well as in a system of rotating cylinders for Couette and Taylor-vortex flow. Different biomaterials are used. The activation, adhesion and aggregation are quantified using scanning electron microscopy and flow cytometry.
Summary of Results: Regions and flow conditions with a high potential for thrombus growth could be identified. The experiments clearly show the influence of the blood contacting material and governing shear rates. Numerical analysis can explain observed adhesion patterns and the degree of thrombus formation
Increasing Scots pine (Pinus sylvestris L.) mortality has been recently observed in the dry inner valleys of the European Alps. Besides drought, infection with pine mistletoe (Viscum album ssp. austriacum) seems to play an important role in the mortality dynamics of Scots pines, but how mistletoes promote pine decline remains unclear. To verify whether pine mistletoe infection weakens the host via crown degradation, as observed for dwarf mistletoes, we studied the negative effects of pine mistletoe infestation on the photosynthetic tissues and branch growth of pairs of infested and non-infested branches. Pine mistletoe infection leads to crown degradation in its host by reducing the length, the radial increment, the ramification, the needle length and the number of needle years of the infested branches. This massive loss in photosynthetic tissue results in a reduction in primary production and a subsequent decrease in carbohydrate availability. The significant reduction in needle length due to mistletoe infection is an indication for a lower water and nutrient availability in infested branches. Thus, mistletoe infection might lead to a decrease in the availability of water and carbohydrates, the two most important growth factors, which are already shortened due to the chronic drought situation in the area. Therefore, pine mistletoe increases the risk of drought-induced mortality of its host when growing in a xeric environment.
Solar sails provide ignificant advantages over other low-thrust propulsion systems because they produce thrust by the momentum exchange from solar radiation pressure (SRP) and thus do not consume any propellant.The force exerted on a very thin sail foil basically depends on the light incidence angle. Several analytical SRP force models that describe the SRP force acting on the sail have been established since the 1970s. All the widely used models use constant optical force coefficients of the reflecting sail material. In 2006,MENGALI et al. proposed a refined SRP force model that takes into account the dependancy of the force coefficients on the light incident angle,the sail’s distance from the sun (and thus the sail emperature) and the surface roughness of the sail material [1]. In this paper, the refined SRP force model is compared to the previous ones in order to identify the potential impact of the new model on the predicted capabilities of solar sails in performing low-cost interplanetary space missions. All force models have been implemented within InTrance, a global low-thrust trajectory optimization software utilizing evolutionary neurocontrol [2]. Two interplanetary rendezvous missions, to Mercury and the near-Earth asteroid 1996FG3, are investigated. Two solar sail performances in terms of characteristic acceleration are examined for both scenarios, 0.2 mm/s2 and 0.5 mm/s2, termed “low” and “medium” sail performance. In case of the refined SRP model, three different values of surface roughness are chosen, h = 0 nm, 10 nm and 25 nm. The results show that the refined SRP force model yields shorter transfer times than the standard model.
In this paper, we will provide a feasible mission design for a multiple-rendezvous mission to Jupiter's Trojans. It is based on solar electric propulsion, as being currently used on the DAWN spacecraft, and other flight-proven technology. First, we have selected a set of mission objectives, the prime objective being the detection of water -especially subsurface water -to provide evidence for the Trojans' formation at large solar distances. Based on DAWN and other comparable missions, we have determined suitable payload instruments to achieve these objectives. Afterwards, we have designed a spacecraft that is able to carry the selected payload to the Trojan region and rendezvous successively with three target bodies within a maximum mission duration of 15 years. Accurate low-thrust trajectories have been obtained with a global low-thrust trajectory optimization program (InTrance). During the transfer from Earth to the first target, the spacecraft is propelled by two RIT-22 ion engines from EADS Astrium, whereas a single RIT-15 is used for transfers within the Trojan region to reduce the required power. For power generation, the spacecraft uses a multi-junction solar array that is supported by concentrators. To achieve moderate mission costs, we have restricted the launch mass to a maximum of 1600 kg, the maximum interplanetary injection capability of a Soyuz/Fregat launcher. Our final layout has a mass of 1400 kg, yielding a margin of about 14%. Nestor (a member of the L4-population) was determined as the first mission target. It can be reached within 4.6 years from launch. The fuel mass ratio for this transfer is about 35%. The stay time at Nestor is 1.2 years. Eurymedon was selected as the second target (transfer time 3.5 years, stay time 3.0 years) and Irus as the third target (transfer time 2.2 years). The transfers within the Trojan L4-population can be accomplished with fuel mass ratios of about 3% for each trajectory leg. Including the stay times in orbit around the targets, the mission can be accomplished within a total duration of about 14.5 years. According to our mission analysis, it is also feasible to fly to the L5-population with similar flight times. It has to be noted that -for a first analysis -we have taken only the named targets into account. Allowing also rendezvous with unnamed objects will very likely decrease the mission duration. Based on a scaling of DAWN's mission costs (due to comparable scientific instruments and mission objectives), and taking into account the longer mission duration and the potential re-use of already developed technology, we have estimated that these three rendezvous can be accomplished with a budget of about 250 Million Euros, i.e. about 25% of ROSETTA's budget.
The chemical imaging sensor is a semiconductor-based chemical sensor that can visualize the two-dimensional distribution of specific ions or molecules in the solution. In this study, we developed a miniaturized chemical imaging sensor system with an OLED display panel as a light source that scans the sensor plate. In the proposed configuration, the display panel is placed directly below the sensor plate and illuminates the back surface. The measured area defined by illumination can be arbitrarily customized to fit the size and the shape of the sample to be measured. The waveform of the generated photocurrent, the currentvoltage characteristics and the pH sensitivity were investigated and pH imaging with this miniaturized system was demonstrated.
Purpose: Image analysis by the retinal vessel analyzer (RVA) observes retinal vessels in their dynamic state online noninvasively along a chosen vessel segment. It has been found that high-frequency diameter changes in the retinal artery blood column along the vessel increase significantly in anamnestically healthy volunteers with increasing age and in patients with glaucoma during vascular dilation. This study was undertaken to investigate whether longitudinal sections of the retinal artery blood column are altered in systemic hypertension.
Methods: Retinal arteries of 15 untreated patients with essential arterial hypertension (age, 50.9 ± 11.9 years) and of 15 age-matched anamnestically healthy volunteers were examined by RVA. After baseline assessment, a monochromatic luminance flicker (530–600 nm; 12.5 Hz; 20 s) was applied to evoke retinal vasodilation. Differences in amplitude and frequency of spatial artery blood column diameter change along segments (longitudinal arterial profiles) of 1 mm in length were measured and analyzed using Fourier transformation.
Results: In the control group, average reduced power spectra (ARPS) of longitudinal arterial profiles did not differ when arteries changed from constriction to dilation. In the systemic hypertension group, ARPS during constriction, baseline, and restoration were identical and differed from ARPS during dilation (P < 0.05). Longitudinal arterial profiles in both groups showed significant dissimilitude at baseline and restoration (P < 0.05).
Conclusions: The retinal artery blood column demonstrates microstructural alterations in systemic hypertension and is less irregular along the vessel axis during vessel dilation. These microstructural changes may be an indication of alterations in vessel wall rigidity, vascular endothelial function, and smooth muscle cells in this disease, leading to impaired perfusion and regulation.
In steps of the production chain of carbide inserts, such as unloading or packaging, the conformity test of the insert type is done manually, which causes a statistic increase of errors due to monotony and fatigue of the worker and the wide variety of the insert types. A machine vision system is introduced that captures digital frames of the inserts in the production line, analyses inspects automatically and measures four quality features: coating colour, edge radius, plate shape and chip-former geometry. This new method has been tested on several inserts of different types and has shown that the prevalent insert types can be inspected and robustly classified in real production environment and therefore improves the manufacturing automation.
This paper develops an investment/pricing model for the deployment of basic broadband networks which, along with other applications, is applicable to public–private partnership projects. In particular, a new investment model is suggested to be used for finance deployment over a longer term by enabling both private and public investors to participate in the roll-out of next generation access (NGA) infrastructure. This so-called “long-term risk sharing concept” has several notable benefits compared with the traditional regulatory approach. Above all, the model enables both private operators and public authorities to share the risk of investing in NGA infrastructure. Thus the model offers a way for public authorities to achieve a timely and countrywide roll-out of NGA networks, including in areas where NGA investment would otherwise not occur.
Lightning safety guidelines
(2010)
This paper introduces lightning to the layman, noting the right behaviour in front of thunderstorms as well as protective measures against lightning. It also contributes to the prevention of lightning injuries and damages. This report was prepared by the authors inside the AHG1 Group for IEC TC81 (Lightning Protection).
A new approach for a label-free electrical detection of DNA hybridization and denaturation using an array of individually addressable field-effect nanoplate SOI (silicon-on-insulator) capacitors functionalized with gold nanoparticles is presented. By using a constant-capacitance measuring setup in a differential mode, signal changes of ∼110 mV and ∼70 mV have been registered after the DNA hybridization and denaturation events, respectively.
• Most of the edible forest mushrooms are mycorrhizal and depend on carbohydrates produced by the associated trees. Fruiting patterns of these fungi are not yet fully understood since climatic factors alone do not completely explain mushroom occurrence.
• The objective of this study was to retrospectively find out if changing tree growth following an increment thinning has influenced the diversity patterns and productivity of associated forest mushrooms in the fungus reserve La Chanéaz, Switzerland.
• The results reveal a clear temporal relationship between the thinning, the growth reaction of trees and the reaction of the fungal community, especially for the ectomycorrhizal species. The tree-ring width of the formerly suppressed beech trees and the fruit body number increased after thinning, leading to a significantly positive correlation between fruit body numbers and tree-ring width.
• Fruit body production was influenced by previous annual tree growth, the best accordance was found between fruit body production and the tree-ring width two years previously.
• The results support the hypothesis that ectomycorrhizal fruit body production must be linked with the growth of the associated host trees. Moreover, the findings indicate the importance of including mycorrhizal fungi as important players when discussing a tree as a carbon source or sink.
Background: One of the most prominent neurobiological models of alexithymia assumes an altered function of the anterior cingulate cortex (ACC) as the crucial neural correlate of alexithymia. So far functional imaging studies have yielded inconclusive results. Therefore, we tested this hypothesis in healthy alexithymics and nonalexithymics in an event-related fMRI study.
Methods: Thirty high- and 30 low-alexithymic right-handed male subjects (selected by the 20-item Toronto Alexithymia Scale, TAS-20) were investigated with event-related fMRI using a picture viewing paradigm. The stimuli consisted of happy, fearful and neutral facial expressions (Ekman-Friesen) as well as positive, negative and neutral pictures from the International Affective Picture System.
Results: Contrasting the high-alexithymic with the low-alexithymic group we observed increased activation of the supragenual ACC for different emotional valences as well as for different emotional stimuli. Moreover, there was a positive correlation of the ACC with the individual TAS-20 scores but no correlations with the individual Beck Depression Inventory scores. Additionally, there was no difference in activity of the amygdala.
Conclusions: We demonstrated that the supragenual ACC is constantly activated more strongly in alexithymic subjects and that this activation is related to the symptoms of alexithymia and not to associated symptoms such as depression. Therefore, our findings support the hypothesis of an altered function of the ACC in alexithymia.
Dexamethasone (DEX) is a potent and widely used anti-inflammatory and immunosuppressant glucocorticoid. It can bind and activate the pregnane X receptor (PXR), which plays a critical role as xenobiotic sensor in mammals to induce the expression of many enzymes, including cytochromes P450 in the CYP3A family. This induction results in its own metabolism. We have used a series of transgenic mouse lines, including a novel, improved humanized PXR line, to compare the induction profile of PXR-regulated drug-metabolizing enzymes after DEX administration, as well as looking at hepatic responses to rifampicin (RIF). The new humanized PXR model has uncovered further intriguing differences between the human and mouse receptors in that RIF only induced Cyp2b10 in the new humanized model. DEX was found to be a much more potent inducer of Cyp3a proteins in wild-type mice than in mice humanized for PXR. To assess whether PXR is involved in the detoxification of DEX in the liver, we analyzed the consequences of high doses of the glucocorticoid on hepatotoxicity on different PXR genetic backgrounds. We also studied these effects in an additional mouse model in which functional mouse Cyp3a genes have been deleted. These strains exhibited different sensitivities to DEX, indicating a protective role of the PXR and CYP3A proteins against the hepatotoxicity of this compound.
The chemical imaging sensor is a semiconductor-based chemical sensor that can visualize the spatial distribution of chemical species. For the practical application of this sensor, artifacts in the chemical images due to defects of the semiconductor substrate and contamination of the sensing surface etc. have been a major problem. An image correction method was developed to eliminate the influence of nonuniformity of individual sensor plate.
Hybrid control for autonomous systems — Integrating learning, deliberation and reactive control
(2010)
Mouse nongenotoxic hepatocarcinogens phenobarbital (PB) and chlordane induce hepatomegaly characterized by hypertrophy and hyperplasia. Increased cell proliferation is implicated in the mechanism of tumor induction. The relevance of these tumors to human health is unclear. The xenoreceptors, constitutive androstane receptors (CARs), and pregnane X receptor (PXR) play key roles in these processes. Novel “humanized” and knockout models for both receptors were developed to investigate potential species differences in hepatomegaly. The effects of PB (80 mg/kg/4 days) and chlordane (10 mg/kg/4 days) were investigated in double humanized PXR and CAR (huPXR/huCAR), double knockout PXR and CAR (PXRKO/CARKO), and wild-type (WT) C57BL/6J mice. In WT mice, both compounds caused increased liver weight, hepatocellular hypertrophy, and cell proliferation. Both compounds caused alterations to a number of cell cycle genes consistent with induction of cell proliferation in WT mice. However, these gene expression changes did not occur in PXRKO/CARKO or huPXR/huCAR mice. Liver hypertrophy without hyperplasia was demonstrated in the huPXR/huCAR animals in response to both compounds. Induction of the CAR and PXR target genes, Cyp2b10 and Cyp3a11, was observed in both WT and huPXR/huCAR mouse lines following treatment with PB or chlordane. In the PXRKO/CARKO mice, neither liver growth nor induction of Cyp2b10 and Cyp3a11 was seen following PB or chlordane treatment, indicating that these effects are CAR/PXR dependent. These data suggest that the human receptors are able to support the chemically induced hypertrophic responses but not the hyperplastic (cell proliferation) responses. At this time, we cannot be certain that hCAR and hPXR when expressed in the mouse can function exactly as the genes do when they are expressed in human cells. However, all parameters investigated to date suggest that much of their functionality is maintained.
High Spatial Resolution 3D MRI of the Larynx Using a Dedicated TX/RX Phased Array Coil at 7.0T
(2010)
MRI holds great potential for elucidating laryngeal and vocal fold anatomy together with the assessment of physiological processes associated in human phonation. However, MRI of human phonation remains very challenging due to the small size of the targeted structures, interfering signal from fat, air between the vocal folds and surrounding muscles and physiological motion. These anatomical/physiological constraints translate into stringent technical requirements in balancing, scan time, image contrast, immunity to physiological motion, temporal resolution and spatial resolution. Motivated by these challenges and limitations this study is aiming at translating the sensitivity gain at ultra-high magnetic fields for enhanced high spatial resolution 3D imaging of the larynx and vocal tract. To approach this goal a dedicated two channel TX/RX larynx coil is being proposed.