G-protein-coupled receptor solubilization and purification for biophysical analysis and functional studies, in the total absence of detergent

G-protein coupled receptors (GPCRs) constitute the largest class of membrane proteins and are a major drug target. A serious obstacle to studying GPCR structure/function characteristics is the requirement to extract the receptors from their native environment in the plasma membrane, coupled with the inherent instability of GPCRs in the detergents required for their solubilization. In the present study, we report the first solubilization and purification of a functional GPCR [human adenosine A2A receptor (A2AR)], in the total absence of detergent at any stage, by exploiting spontaneous encapsulation by styrene maleic acid (SMA) co-polymer direct from the membrane into a nanoscale SMA lipid particle (SMALP). Furthermore, the A2AR-SMALP, generated from yeast (Pichia pastoris) or mammalian cells, exhibited increased thermostability (∼5°C) compared with detergent [DDM (n-dodecyl-β-D-maltopyranoside)]-solubilized A2AR controls. The A2AR-SMALP was also stable when stored for prolonged periods at 4°C and was resistant to multiple freeze-thaw cycles, in marked contrast with the detergent-solubilized receptor. These properties establish the potential for using GPCR-SMALP in receptor-based drug discovery assays. Moreover, in contrast with nanodiscs stabilized by scaffold proteins, the non-proteinaceous nature of the SMA polymer allowed unobscured biophysical characterization of the embedded receptor. Consequently, CD spectroscopy was used to relate changes in secondary structure to loss of ligand binding ([³H]ZM241385) capability. SMALP-solubilization of GPCRs, retaining the annular lipid environment, will enable a wide range of therapeutic targets to be prepared in native-like state to aid drug discovery and understanding of GPCR molecular mechanisms.

Economical analysis for investment on measuring systems

Metrology processes contribute to entire manufacturing systems that can have a considerable impact on financial investment in coordinate measuring systems. However, there is a lack of generic methodologies to quantify their economical value in today’s industry. To solve this problem, a mathematical model is proposed in this paper by statistical deductive reasoning. This is done through defining the relationships between Process Capability Index, measurement uncertainty and tolerance band. The correctness of the mathematical model is proved by a case study. Finally, several comments and suggestions on evaluating and maximizing the benefits of metrology investment are given.

Numerical analysis of artificial neural network and volterra-based nonlinear equalizers for coherent optical OFDM

One major drawback of coherent optical orthogonal frequency-division multiplexing (CO-OFDM) that hitherto remains unsolved is its vulnerability to nonlinear fiber effects due to its high peak-to-average power ratio. Several digital signal processing techniques have been investigated for the compensation of fiber nonlinearities, e.g., digital back-propagation, nonlinear pre- and post-compensation and nonlinear equalizers (NLEs) based on the inverse Volterra-series transfer function (IVSTF). Alternatively, nonlinearities can be mitigated using nonlinear decision classifiers such as artificial neural networks (ANNs) based on a multilayer perceptron. In this paper, ANN-NLE is presented for a 16QAM CO-OFDM system. The capability of the proposed approach to compensate the fiber nonlinearities is numerically demonstrated for up to 100-Gb/s and over 1000km and compared to the benchmark IVSTF-NLE. Results show that in terms of Q-factor, for 100-Gb/s at 1000km of transmission, ANN-NLE outperforms linear equalization and IVSTF-NLE by 3.2dB and 1dB, respectively.

Is more capability always beneficial for firm performance? Market orientation, core business process capabilities, and business environment

This study examines the role of capabilities in core marketing-related business processes–product development management (PDM), supply chain management (SCM) and customer relationship management (CRM)–in translating a firm’s market orientation (MO) into firm performance. The study is the first to examine the interplay of all three business process capabilities simultaneously, while investigating how environmental conditions moderate their performance effects. A moderated mediation analysis of 468 product-focused firms finds that PDM and CRM process capabilities play important mediating roles, whereas SCM process capability does not mediate the relationship between MO and performance. However, the relative importance of the capabilities as mediators varies along the degree of environmental turbulence, and under certain conditions, an increase in the level of business process capability may even turn detrimental.

Simulation and experimental analysis of a brushless electrically excited synchronous machine with a hybrid rotor

Electrically excited synchronous machines with brushes and slip rings are popular but hardly used in inflammable and explosive environments. This paper proposes a new brushless electrically excited synchronous motor with a hybrid rotor. It eliminates the use of brushes and slip rings so as to improve the reliability and cost-effectiveness of the traction drive. The proposed motor is characterized with two sets of stator windings with two different pole numbers to provide excitation and drive torque independently. This paper introduces the structure and operating principle of the machine, followed by the analysis of the air-gap magnetic field using the finite-element method. The influence of the excitation winding's pole number on the coupling capability is studied and the operating characteristics of the machine are simulated. These are further examined by the experimental tests on a 16 kW prototype motor. The machine is proved to have good static and dynamic performance, which meets the stringent requirements for traction applications.

Development of a technological innovation capability assessment model:a case study of manufacturing SMEs in Sialkot, Pakistan

Small and Medium Enterprises (SMEs) play an important part in the economy of any country. Initially, a flat management hierarchy, quick response to market changes and cost competitiveness were seen as the competitive characteristics of an SME. Recently, in developed economies, technological capabilities (TCs) management- managing existing and developing or assimilating new technological capabilities for continuous process and product innovations, has become important for both large organisations and SMEs to achieve sustained competitiveness. Therefore, various technological innovation capability (TIC) models have been developed at firm level to assess firms‘ innovation capability level. These models output help policy makers and firm managers to devise policies for deepening a firm‘s technical knowledge generation, acquisition and exploitation capabilities for sustained technological competitive edge. However, in developing countries TCs management is more of TCs upgrading: acquisitions of TCs from abroad, and then assimilating, innovating and exploiting them. Most of the TIC models for developing countries delineate the level of TIC required as firms move from the acquisition to innovative level. However, these models do not provide tools for assessing the existing level of TIC of a firm and various factors affecting TIC, to help practical interventions for TCs upgrading of firms for improved or new processes and products. Recently, the Government of Pakistan (GOP) has realised the importance of TCs upgrading in SMEs-especially export-oriented, for their sustained competitiveness. The GOP has launched various initiatives with local and foreign assistance to identify ways and means of upgrading local SMEs capabilities. This research targets this gap and developed a TICs assessment model for identifying the existing level of TIC of manufacturing SMEs existing in clusters in Sialkot, Pakistan. SME executives in three different export-oriented clusters at Sialkot were interviewed to analyse technological capabilities development initiatives (CDIs) taken by them to develop and upgrade their firms‘ TCs. Data analysed at CDI, firm, cluster and cross-cluster level first helped classify interviewed firms as leader, follower and reactor, with leader firms claiming to introduce mostly new CDIs to their cluster. Second, the data analysis displayed that mostly interviewed leader firms exhibited ‗learning by interacting‘ and ‗learning by training‘ capabilities for expertise acquisition from customers and international consultants. However, these leader firms did not show much evidence of learning by using, reverse engineering and R&D capabilities, which according to the extant literature are necessary for upgrading existing TIC level and thus TCs of firm for better value-added processes and products. The research results are supported by extant literature on Sialkot clusters. Thus, in sum, a TIC assessment model was developed in this research which qualitatively identified interviewed firms‘ TIC levels, the factors affecting them, and is validated by existing literature on interviewed Sialkot clusters. Further, the research gives policy level recommendations for TIC and thus TCs upgrading at firm and cluster level for targeting better value-added markets.

Technological capability development in China's energy service company industry

Technological capability (TC) plays a strategic role in the competitive advantage of not only individual corporate entities but also entire industries. This paper investigates the crucial factors that affect technological capability development by Energy Service Companies (ESCOs) in China. It identifies how differently sized ESCOs make progress in developing TCs. Through looking at the successes achieved by developed countries in the field of energy conservation, ESCOs are able to improve energy efficiency and reduce emissions and are deemed to provide an effective means of conserving energy in China. Existing literature indicates that limited TC levels of are one of the crucial barriers facing Chinese ESCOs. Through investigating three different sizes of Chinese ESCO - small, medium-sized and large - this paper provides a framework to present the idea that Chinese ESCOs' TC development is affected by four key internal and external capabilities: management capability, investment capability, innovation capability and linkage capability. Through comparative analysis, the paper establishes that small and medium-sized private ESCOs are mainly affected by investment and linkage capabilities. Large state-owned ESCOs are mainly affected by innovation and management capability. In addition, all three types of ESCO exhibit a strong desire to develop their technological capability, but small and medium-sized ESCOs exhibit a stronger desire to conduct research and development (R&D) than large ESCOs, whilst large ESCOs prefer to increase their technical reserves through acquisition. This paper identifies factors that affect Chinese ESCOs' TC, but it does intend to address the problem of how to reduce the negative effects of limited TC or the question of how to improve the TC development of Chinese ESCOs effectively. This paper contributes to the field of TC development in the ESCO industry.