A high performance liquid chromatography and electrospray ionization mass spectrometry method for the analysis of the natural medicine, Forsythia suspensa

Samples of Forsythia suspensa from raw (Laoqiao) and ripe (Qingqiao) fruit were analyzed with the use of HPLC-DAD and the EIS-MS techniques. Seventeen peaks were detected, and of these, twelve were identified. Most were related to the glucopyranoside molecular fragment. Samples collected from three geographical areas (Shanxi, Henan and Shandong Provinces), were discriminated with the use of hierarchical clustering analysis (HCA), discriminant analysis (DA), and principal component analysis (PCA) models, but only PCA was able to provide further information about the relationships between objects and loadings; eight peaks were related to the provinces of sample origin. The supervised classification models-K-nearest neighbor (KNN), least squares support vector machines (LS-SVM), and counter propagation artificial neural network (CP-ANN) methods, indicated successful classification but KNN produced 100% classification rate. Thus, the fruit were discriminated on the basis of their places of origin.

Natural mapping and intuitive interaction in videogames

Videogame control interfaces continue to evolve beyond their traditional roots, with devices encouraging more natural forms of interaction growing in number and pervasiveness. Yet little is known about their true potential for intuitive use. This paper proposes methods to leverage existing intuitive interaction theory for games research, specifically by examining different types of naturally mapped control interfaces for videogames using new measures for previous player experience. Three commercial control devices for a racing game were categorised using an existing typology, according to how the interface maps physical control inputs with the virtual gameplay actions. The devices were then used in a within-groups (n=64) experimental design aimed at measuring differences in intuitive use outcomes. Results from mixed design ANOVA are discussed, along with implications for the field.

Versatility of polymethacrylate monoliths for chromatographic purification of biomolecules

Polymethacrylate monoliths, specifically poly(glycidyl methacrylate-co-ethylene dimethacrylate) or poly(GMA-co-EDMA) monoliths, are a new generation of chromatographic supports and are significantly different from conventional particle-based adsorbents, membranes, and other monolithic supports for biomolecule purification. Similar to other monoliths, polymethacrylate monoliths possess large pores which allow convective flow of mobile phase and result in high flow rates at reduced pressure drop, unlike particulate supports. The simplicity of the adsorbent synthesis, pH resistance, and the ease and flexibility of tailoring their pore size to that of the target biomolecule are the key properties which differentiate polymethacrylate monoliths from other monoliths. Polymethacrylate monoliths are endowed with reactive epoxy groups for easy functionalization (with anion-exchange, hydrophobic, and affinity ligands) and high ligand retention. In this review, the structure and performance of polymethacrylate monoliths for chromatographic purification of biomolecules are evaluated and compared to those of other supports. The development and use of polymethacrylate monoliths for research applications have grown rapidly in recent times and have enabled the achievement of high through-put biomolecule purification on semi-preparative and preparative scales.

Binding properties of peptidic affinity ligands for plasmid DNA capture and detection

Peptides constructed from α-helical subunits of the Lac repressor protein (LacI) were designed then tailored to achieve particular binding kinetics and dissociation constants for plasmid DNA purification and detection. Surface plasmon resonance was employed for quantification and characterization of the binding of double stranded Escherichia coli plasmid DNA (pUC19) via the lac operon (lacO) to "biomimics" of the DNA binding domain of LacI. Equilibrium dissociation constants (K D), association (k a), and dissociation rates (k d) for the interaction between a suite of peptide sequences and pUC19 were determined. K D values measured for the binding of pUC19 to the 47mer, 27mer, 16mer, and 14mer peptides were 8.8 ± 1.3 × 10 -10 M, 7.2 ± 0.6 × 10 -10 M, 4.5 ± 0.5 × 10 -8 M, and 6.2 ± 0.9 × 10 -6 M, respectively. These findings show that affinity peptides, composed of subunits from a naturally occurring operon-repressor interaction, can be designed to achieve binding characteristics suitable for affinity chromatography and biosensor devices.

Single step purification of plasmid DNA using peptide ligand affinity chromatography

Single step affinity chromatography was employed for the purification of plasmid DNA (pDNA), thus eliminating several steps compared with current commercial purification methods for pDNA. Significant reduction in pDNA production time and cost was obtained. This chromatographic operation employed a peptide-monolith construct to isolate pDNA from Escherichia coli (E. coli) impurities present in a clarified lysate feedstock. Mild conditions were applied to avoid any degradation of pDNA. The effect of some important parameters on pDNA yield was also evaluated with the aim of optimising the affinity purification of pDNA. The results demonstrate that 81% of pDNA was recovered and contaminating gDNA, RNA and protein were removed below detectable levels. © 2008 Elsevier B.V. All rights reserved.

Binding mechanism of affinity ligands for purification of plasmid DNA

Plasmid DNA for therapeutic and vaccination purposes must be highly purified. The high selectivity of affinity chromatography makes it ideal for the isolation of pDNA from complex biological feed stocks. Affinity chromatography makes use of the biological function and/or individual chemical structure of the interacting molecules. However, the success of any affinity purification protocol is dependent on the availability of suitable ligands. In this study, surface plasmon resonance (SPR) based Biacore system has been employed for the detection and quantification of the binding between lac operon (lacO) sequence contained in a pDNA and synthetic peptides based on the DNA-binding domain of the lac repressor protein, lad. The equilibrium dissociation constant (K D) and association and dissociation rate constants (ka, kd) for the interaction between plasmid DNA and designed peptides were determined.

Affinity purification of plasmid DNA directly from crude bacterial cell lysates

We have shown previously that a sequence-specific DNA-binding protein based on the Lac repressor protein can isolate pre-purified DNA efficiently from simple buffer solution but our attempts to purify plasmids directly from crude starting materials were disappointing with unpractically low DNA yields. We have optimized tbe procedure and present a simple affinity methodology whereby plasmid DNA is purified directly by mixing two crude cell lysates, one cell lysate containing the plasmid and the other the protein affinity ligand, without the need for treatment by RNaseA. After IMAC chromatography, high purity supercoiled DNA is recovered in good yields of 100-150 μg plasmid per 200 mL shake flask culture. Moreover, the resulting DNA is free from linear or open-circular plasmid DNA, genomic DNA, RNA, and protein, to the limits of our detection. Furthermore, we show that lyophilized affinity ligand can be stored at room temperature and re-hydrated for use when required.

Affinity adsorption of plasmid DNA

The development of a protein-mediated dual functional affinity adsorption of plasmid DNA is described in this work. The affinity ligand for the plasmid DNA comprises a fusion protein with glutathione-S-transferase (GST) as the fusion partner with a zinc finger protein. The protein ligand is first bound to the adsorbent by affinity interaction between the GST moeity and gluthathione that is covalently immobilized to the base matrix. The plasmid binding is then enabled via the zinc finger protein and a specific nucleotide sequence inserted into the DNA. At lower loadings, the binding of the DNA onto the Fractogel, Sepharose, and Streamline matrices was 0.0078 ± 0.0013, 0.0095 ± 0.0016, and 0.0080 ± 0.0006 mg, respectively, to 50 μL of adsorbent. At a higher DNA challenge, the corresponding amounts were 0.0179 ± 0.0043, 0.0219 ± 0.0035, and 0.0190 ± 0.0041 mg, respectively. The relatively constant amounts bound to the three adsorbents indicated that the large DNA molecule was unable to utilize the available zinc finger sites that were located in the internal pores and binding was largely a surface adsorption phenomenon. Utilization of the zinc finger binding sites was shown to be highest for the Fractogel adsorbent. The adsorbed material was eluted with reduced glutathione, and the eluted efficiency for the DNA was between 23% and 27%. The protein elution profile appeared to match the adsorption profiles with significantly higher recoveries of bound GST-zinc finger protein.

Composing an email : social interaction in a preschool classroom

The chapter contributes understandings of digitally mediated interactions in early childhood classrooms. Ethnomethodological and conversation analysis approaches are used to analyse a video-recorded episode of children and teacher composing an email in a preschool classroom. In their talk we find how the teacher directs the children to what counts procedurally, such as the components of an email, and the teacher’s moral work in producing a culturally correct way form of personal communication. Such considerations of situated examples can encourage investigations of digital practices that extend beyond operational skills to broader understandings of digital practices as cultural and situated activities.

Mobile interaction design approaches for reducing domestic food waste

This thesis is a trans-disciplinary study of domestic food waste in Australia. Firstly, it examines why consumers are prone to waste food. Secondly, it explores several situated design interventions to reduce domestic food waste by informing consumer food supply and location awareness, and improving the level of food literacy among consumers. The thesis outcomes have implications for academic and industry domains within the fields of Human-Computer Interaction, urban informatics, environmental sustainability, food security and public health.

Social interaction and the role of empathy in information and knowledge management

This article provides a general review of the literature on the nature and role of empathy in social interaction for information professionals working in a variety of information and knowledge environments. Relational agency theory (Edwards, 2005) is used asa framework to re-conceptualize education for empathic social interaction between information professionals and their clients. Past, present and future issues relevant to empathic interaction in information and knowledge management are discussed in the context of three shifts identified from the literature: (a) the continued increase in communication channels, both physical and virtual, for reference, information and re-search services, (b) the transition from the information age to the conceptual age and(c) the growing need for understanding of the affective paradigm in the information and knowledge professions. Findings from the literature review on the relationships between empathy and information behavior, social networking, knowledge management and information and knowledge services are presented. Findings are discussed in relation to the development of guidelines for the affective education and training of information and knowledge professionals and the potential use of virtual learning software such as Second Life in developing empathic communication skills

Refactoring Service Interface to Support Artefact-centric Service interaction

The growth of APIs and Web services on the Internet, especially through larger enterprise systems increasingly being leveraged for Cloud and software-as-a-service opportunities, poses challenges for improving the efficiency of integration with these services. Interfaces of enterprise systems are typically larger, more complex and overloaded, with single operations having multiple data entities and parameter sets, supporting varying requests, and reflecting versioning across different system releases, compared to fine-grained operations of contemporary interfaces. We propose a technique to support the refactoring of service interfaces by deriving business entities and their relationships. In this paper, we focus on the behavioural aspects of service interfaces, aiming to discover the sequential dependencies of operations (otherwise known as protocol extraction) based on the entities and relationships derived. Specifically, we propose heuristics according to these relationships, and in turn, deriving permissible orders in which operations are invoked. As a result of this, service operations can be refactored on business entity CRUD lines, with explicit behavioural protocols as part of an interface definition. This supports flexible service discovery, composition and integration. A prototypical implementation and analysis of existing Web services, including those of commercial logistic systems (Fedex), are used to validate the algorithms proposed through the paper.