Identification of a cyclase-associated protein (CAP) homologue in Dictyostelium discoideum and characterization of its interaction with actin

In search for novel actin binding proteins in Dictyostelium discoideum we have isolated a cDNA clone coding for a protein of approximately 50 kDa that is highly homologous to the class of adenylyl cyclase-associated proteins (CAP). In Saccharomyces cerevisiae the amino-terminal part of CAP is involved in the regulation of the adenylyl cyclase whereas the loss of the carboxyl-terminal domain results in morphological and nutritional defects. To study the interaction of Dictyostelium CAP with actin, the complete protein and its amino-terminal and carboxyl-terminal domains were expressed in Escherichia coli and used in actin binding assays. CAP sequestered actin in a Ca2+ independent way. This activity was localized to the carboxyl-terminal domain. CAP and its carboxyl-terminal domain led to a fluorescence enhancement of pyrene-labeled G-actin up to 50% indicating a direct interaction, whereas the amino-terminal domain did not enhance. In polymerization as well as in viscometric assays the ability of the carboxyl-terminal domain to sequester actin and to prevent F-actin formation was approximately two times higher than that of intact CAP. The sequestering activity of full length CAP could be inhibited by phosphatidylinositol 4,5-bisphosphate (PIP2), whereas the activity of the carboxyl-terminal domain alone was not influenced, suggesting that the amino-terminal half of the protein is required for the PIP2 modulation of the CAP function. In profilin-minus cells the CAP concentration is increased by approximately 73%, indicating that CAP may compensate some profilin functions in vivo. In migrating D. discoideum cells CAP was enriched at anterior and posterior plasma membrane regions. Only a weak staining of the cytoplasm was observed. In chemotactically stimulated cells the protein was very prominent in leading fronts. The data suggest an involvement of D. discoideum CAP in microfilament reorganization near the plasma membrane in a PIP2-regulated manner.     

A New Research Infrastructure for Investigating Flow Hydraulics and Process Equipment at Critical Fluid Properties

An optimized equipment design for natural gas processing and liquefaction plants becomes increasingly difficult with changing process conditions: Particularly low values of surface tension create rising challenges on the design of phase separators and column internals. The TERESA test rig at HZDR was designed to allow the investigation of multiphase thermohydraulics and phase separator performance under critical fluid properties in industrial dimensions. A versatile pipe test section is available in DN200 and equipment internals may be tested in a sectional DN300/DN500 test separator. The applied test fluid shows a high vapor-liquid density difference between 1470 and 940 kg m⁻³, viscosity as low as 0.12 mm²s⁻¹, and surface tension down to 1.3 mN m⁻¹. Volumetric liquid and vapor flow rates may be varied up to 9 and 530 m³h⁻¹ in the test rig, respectively.     

SOA-enabled compliance management: instrumenting, assessing, and analyzing service-based business processes

Facilitating compliance management, that is, assisting a company’s management in conforming to laws, regulations, standards, contracts, and policies, is a hot but non-trivial task. The service-oriented architecture (SOA) has evolved traditional, manual business practices into modern, service-based IT practices that ease part of the problem: the systematic definition and execution of business processes. This, in turn, facilitates the online monitoring of system behaviors and the enforcement of allowed behaviors—all ingredients that can be used to assist compliance management on the fly during process execution. In this paper, instead of focusing on monitoring and runtime enforcement of rules or constraints, we strive for an alternative approach to compliance management in SOAs that aims at assessing and improving compliance. We propose two ingredients: (i) a model and tool to design compliant service-based processes and to instrument them in order to generate evidence of how they are executed and (ii) a reporting and analysis suite to create awareness of a company’s compliance state and to enable understanding why and where compliance violations have occurred. Together, these ingredients result in an approach that is close to how the real stakeholders—compliance experts and auditors—actually assess the state of compliance in practice and that is less intrusive than enforcing compliance.     

Real-time hierarchical stereo Visual SLAM in large-scale environments

In this paper we present a new real-time hierarchical (topological/metric) Visual SLAM system focusing on the localization of a vehicle in large-scale outdoor urban environments. It is exclusively based on the visual information provided by a cheap wide-angle stereo camera. Our approach divides the whole map into local sub-maps identified by the so-called fingerprints (vehicle poses). At the sub-map level (low level SLAM), 3D sequential mapping of natural landmarks and the robot location/orientation are obtained using a top-down Bayesian method to model the dynamic behavior. A higher topological level (high level SLAM) based on fingerprints has been added to reduce the global accumulated drift, keeping real-time constraints. Using this hierarchical strategy, we keep the local consistency of the metric sub-maps, by mean of the EKF, and global consistency by using the topological map and the MultiLevel Relaxation (MLR) algorithm. Some experimental results for different large-scale outdoor environments are presented, showing an almost constant processing time.     

Making it easier for older people to talk to smart homes: the effect of early help prompts

It is well known that help prompts shape how users talk to spoken dialogue systems. This study investigated the effect of help prompt placement on older users’ interaction with a smart home interface. In the dynamic help condition, help was only given in response to system errors; in the inherent help condition, it was also given at the start of each task. Fifteen older and sixteen younger users interacted with a smart home system using two different scenarios. Each scenario consisted of several tasks. The linguistic style users employed to communicate with the system (interaction style) was measured using the ratio of commands to the overall utterance length (keyword ratio) and the percentage of content words in the user’s utterance that could be understood by the system (shared vocabulary). While the timing of help prompts did not affect the interaction style of younger users, it was early task-specific help supported older users in adapting their interaction style to the system’s capabilities. Well-placed help prompts can significantly increase the usability of spoken dialogue systems for older people.     

Dual-plane ultrafast limited-angle electron beam x-ray tomography

Electron beam x-ray tomography is an imaging technique, which can provide cross-sectional images of an object of interest with about 1 mm spatial resolution at frame rates of up to 10,000 frames per second. As a non-intrusive method it is especially suited for studying multiphase flows. For this purpose we devised an experimental limited-angle scan setup which utilizes linear beam deflection to generate radiographic projections. This setup was employed in the study of gas–liquid flow in an experimental flow loop operated at different liquid and gas flow rates. Electron-beam tomography images were compared with image data of a wire-mesh sensor. The latter is a fast but intrusive imaging device which is commonly used in gas–liquid flow imaging and achieves comparable frame rates but at lower spatial resolution. As a novelty we implemented a dual-plane limited-angle electron beam x-ray tomography which allows us to gain information about the phase velocities using cross-correlation data analysis.     

提供恒定发光度的LED驱动器

图1所示电路的输出电流在1.2～1.5V的输入电压范围内几乎是恒定的,并对晶体管的增益变化不敏感.晶体管Q1和Q2组成一个非稳态触发器.R1和C确定Q2的导通时间.在Q2导通期间,Q1截止,Q1的基极电压和电感器L中的电流逐渐升高.当Q1的基极电压达到大约0.6V时,Q1导通,而Q2截止.这种转换在电感器L中引起"逆转"动作.电感器两端的电压极性相反,存储在电感器中的能量以下降的脉冲电流形式传送给LED.在逆转期间,LED两端的电压几乎是恒定的.     

HF单边带解调器

图1所示电路用于补充“设计实例(Design Idea)”栏目以前的一篇文章渗考文献1)。图中的调制器使用了一个移相网络,把一个低频的(声频)信号分成同相和90°相差(正交)的两个分量。这里的移相网络比其他移相电路优越之处在于,它产生的移相误     

Miniature conductivity wire-mesh sensor for gas-liquid two-phase flow measurement

A miniature conductivity wire-mesh sensor for gas-liquid two-phase flow measurement in small channels is presented. The sensor design is similar to the conventional wire-mesh sensor for larger flow cross sections with wire electrodes stretched across the flow channel in two adjacent planes and with perpendicular wire orientation between planes. Conductivity measurement is performed by special electronics which consecutively applies bipolar voltage pulse excitation to the sender wires and measures electrical current flow in the wire crossings at the receiver wires. The new design is based on printed circuit board technology. A prototypical sensor made of 2×16 stainless steel wires each of 50 μm diameter was manufactured and applied to two-phase flow measurement inside the mixing chamber of an effervescent atomizer. Accuracy of the sensor was studied for static liquid distributions using microphotography and for dynamic two-phase flow by comparison of wire-mesh sensor data with radial gas fraction profiles obtained from X-ray microtomography measurements.     

Importance of a flexible hinge near the motor domain in kinesin-driven motility

Conventional kinesin is a molecular motor consisting of an N-terminal catalytic motor domain, an extended stalk and a small globular C-terminus. Whereas the structure and function of the catalytic motor domain has been investigated, little is known about the function of domains outside the globular head. A short coiled-coil region adjacent to the motor domain, termed the neck, is known to be important for dimerization and may be required for kinesin processivity. We now provide evidence that a helix-disrupting hinge region (hinge 1) that separates the neck from the first extended coiled-coil of the stalk plays an essential role in basic motor activity. A fast fungal kinesin from Syncephalastrum racemosum was used for these studies. Deletion, substitution by a coiled-coil and truncation of the hinge 1 region all reduce motor speed and uncouple ATP turnover from gliding velocity. Insertion of hinge 1 regions from two conventional kinesins, Nkin and DmKHC, fully restores motor activity, whereas insertion of putative flexible linkers of other proteins does not, suggesting that hinge 1 regions of conventional kinesins can functionally replace each other. We suggest that this region is essential for kinesin movement in its promotion of chemo-mechanical coupling of the two heads and therefore the functional motor domain should be redefined to include not only the catalytic head but also the adjacent neck and hinge 1 domains.