Single molecule microscopy in 3D cell cultures and tissues

From the onset of the first microscopic visualization of single fluorescent molecules in living cells at the beginning of this century, to the present, almost routine application of single molecule microscopy, the method has well-proven its ability to contribute unmatched detailed insight into the heterogeneous and dynamic molecular world life is composed of. Except for investigations on bacteria and yeast, almost the entire story of success is based on studies on adherent mammalian 2D cell cultures. However, despite this continuous progress, the technique was not able to keep pace with the move of the cell biology community to adapt 3D cell culture models for basic research, regenerative medicine, or drug development and screening. In this review, we will summarize the progress, which only recently allowed for the application of single molecule microscopy to 3D cell systems and give an overview of the technical advances that led to it. While initially posing a challenge, we finally conclude that relevant 3D cell models will become an integral part of the on-going success of single molecule microscopy.

Biomimicking natural nanopatterned topology by 3D laser lithography

Natural nanopatterned surfaces (nNPS) present on insect wings have demonstrated bactericidal activity [1, 2]. Fabricated nanopatterned surfaces (fNPS) derived by characterization of these wings have also shown superior bactericidal activity [2]. However bactericidal NPS topologies vary in both geometry and chemical characteristics of the individual features in different insects and fabricated surfaces, rendering it difficult to ascertain the optimum geometrical parameters underling bactericidal activity. This situation calls for the adaptation of new and emerging techniques, which are capable of fabricating and characterising comparable structures to nNPS from biocompatible materials. In this research, CAD drawn nNPS representing an area of 10 μm x10 μm was fabricated on a fused silica glass by Nanoscribe photonic professional GT 3D laser lithography system using two photon polymerization lithography. The glass was cleaned with acetone and isopropyl alcohol thrice and a drop of IP-DIP photoresist from Nanoscribe GmbH was cast onto the glass slide prior to patterning. Photosensitive IP-DIP resist was polymerized with high precision to make the surface nanopatterns using a 780 nm wavelength laser. Both moving-beam fixedsample (MBFS) and fixed-beam moving-sample (FBMS) fabrication approaches were tested during the fabrication process to determine the best approach for the precise fabrication of the required nanotopological pattern. Laser power was also optimized to fabricate the required fNPS, where this was changed from 3mW to 10mW to determine the optimum laser power for the polymerization of the photoresist for fabricating FNPS...

Enterprise system investments for competitive advantage: An empirical study of Swiss SMEs

Previous research identifies various reasons companies invest in information technology (IT), often as a means to generate value. To add to the discussion of IT value generation, this study investigates investments in enterprise software systems that support business processes. Managers of more than 500 Swiss small and medium-sized enterprises (SMEs) responded to a survey regarding the levels of their IT investment in enterprise software systems and the perceived utility of those investments. The authors use logistic and ordinary least squares regression to examine whether IT investments in two business processes affect SMEs' performance and competitive advantage. Using cluster analysis, they also develop a firm typology with four distinct groups that differ in their investments in enterprise software systems. These findings offer key implications for both research and managerial practice.

Missing pieces in the puzzle of plant microRNAs

Plant microRNAs (miRNAs) are important regulatory switches. Recent advances have revealed many regulatory layers between the two essential processes, miRNA biogenesis and function. However, how these multilayered regulatory processes ultimately control miRNA gene regulation and connects miRNAs and plant responses with the surrounding environment is still largely unknown. In this opinion article, we propose that the miRNA pathway is highly dynamic and plastic. The apparent flexibility of the miRNA pathway in plants appears to be controlled by a number recently identified proteins and poorly characterized signaling cascades. We further propose that altered miRNA accumulation can be a direct consequence of the rewiring of interactions between proteins that function in the miRNA pathway, an avenue that remains largely unexplored.

Laboratory measurement of hydraulic conductivity functions of two unsaturated sandy soils during drying and wetting processes

The importance of applying unsaturated soil mechanics to geotechnical engineering design has been well understood. However, the consumption of time and the necessity for a specific laboratory testing apparatus when measuring unsaturated soil properties have limited the application of unsaturated soil mechanics theories in practice. Although methods for predicting unsaturated soil properties have been developed, the verification of these methods for a wide range of soil types is required in order to increase the confidence of practicing engineers in using these methods. In this study, a new permeameter was developed to measure the hydraulic conductivity of unsaturated soils using the steady-state method and directly measured suction (negative pore-water pressure) values. The apparatus is instrumented with two tensiometers for the direct measurement of suction during the tests. The apparatus can be used to obtain the hydraulic conductivity function of sandy soil over a low suction range (0-10 kPa). Firstly, the repeatability of the unsaturated hydraulic conductivity measurement, using the new permeameter, was verified by conducting tests on two identical sandy soil specimens and obtaining similar results. The hydraulic conductivity functions of the two sandy soils were then measured during the drying and wetting processes of the soils. A significant hysteresis was observed when the hydraulic conductivity was plotted against the suction. However, the hysteresis effects were not apparent when the conductivity was plotted against the volumetric water content. Furthermore, the measured unsaturated hydraulic conductivity functions were compared with predictions using three different predictive methods that are widely incorporated into numerical software. The results suggest that these predictive methods are capable of capturing the measured behavior with reasonable agreement.

Characterisation of two distinctly different processes associated with the electrocrystallization of microcrystals of phase I CuTCNQ (TCNQ= 7, 7, 8, 8-tetracyanoquinodimethane)

Semi-conducting phase I CuTCNQ (TCNQ = 7,7,8,8-tetracyanoquinodimethane), which is of considerable interest as a switching device for memory storage materials, can be electrocrystallized from CH3CN via two distinctly different pathways when TCNQ is reduced to TCNQ˙− in the presence of [Cu(MeCN)4]+. The first pathway, identified in earlier studies, occurs at potentials where TCNQ is reduced to TCNQ˙− and involves a nucleation–growth mechanism at preferred sites on the electrode to produce arrays of well separated large branched needle-shaped phase I CuTCNQ crystals. The second pathway, now identified at more negative potentials, generates much smaller needle-shaped phase I CuTCNQ crystals. These electrocrystallize on parts of the surface not occupied in the initial process and give rise to film-like characteristics. This process is attributed to the reduction of Cu+[(TCNQ˙−)(TCNQ)] or a stabilised film of TCNQ via a solid–solid conversion process, which also involves ingress of Cu+via a nucleation–growth mechanism. The CuTCNQ surface area coverage is extensive since it occurs at all areas of the electrode and not just at defect sites that dominate the crystal formation sites for the first pathway. Infrared spectra, X-ray diffraction, surface plasmon resonance, quartz crystal microbalance, scanning electron microscopy and optical image data all confirm that two distinctly different pathways are available to produce the kinetically-favoured and more highly conducting phase I CuTCNQ solid, rather than the phase II material.

Electrochemically-induced TCNQ/Mn TCNQ 2 (H2O) 2 (TCNQ= 7, 7, 8, 8-Tetracyanoquinodimethane) solid-solid interconversion : two voltammetrically distinct processes that allow selective generation of nanofiber or nanorod network morphologies

Unlike the case with other divalent transition metal M\[TCNQ](2)(H(2)O)(2) (M = Fe, Co, Ni) analogues, the electrochemically induced solid-solid phase interconversion of TCNQ microcrystals (TCNQ = 7,7,8,8-tetracyanoquinodimethane) to Mn\[TCNQ](2)(H(2)O)(2) occurs via two voltammetrically distinct, time dependent processes that generate the coordination polymer in nanofiber or rod-like morphologies. Careful manipulation of the voltammetric scan rate, electrolysis time, Mn(2+)((aq)) concentration, and the method of electrode modification with solid TCNQ allows selective generation of either morphology. Detailed ex situ spectroscopic (IR, Raman), scanning electron microscopy (SEM), and X-ray powder diffraction (XRD) characterization clearly establish that differences in the electrochemically synthesized Mn-TCNQ material are confined to morphology. Generation of the nanofiber form is proposed to take place rapidly via formation and reduction of a Mn-stabilized anionic dimer intermediate, \[(Mn(2+))(TCNQ-TCNQ)(2)(*-)], formed as a result of radical-substrate coupling between TCNQ(*-) and neutral TCNQ, accompanied by ingress of Mn(2+) ions from the aqueous solution at the triple phase TCNQ/electrode/electrolyte boundary. In contrast, formation of the nanorod form is much slower and is postulated to arise from disproportionation of the \[(Mn(2+))(TCNQ-TCNQ)(*-)(2)] intermediate. Thus, identification of the time dependent pathways via the solid-solid state electrochemical approach allows the crystal size of the Mn\[TCNQ](2)(H(2)O)(2) material to be tuned and provides new mechanistic insights into the formation of different morphologies.

Is the sum more than the corporate entrepreneurship parts? the different effects of social capital on innovation, venturing, and renewal processes

Principal Topic Although corporate entrepreneurship is of vital importance for long-term firm survival and growth (Zahra and Covin, 1995), researchers still struggle with understanding how to manage corporate entrepreneurship activities. Corporate entrepreneurship consists of three parts: innovation, venturing, and renewal processes (Guth and Ginsberg, 1990). Innovation refers to the development of new products, venturing to the creation of new businesses, and renewal to redefining existing businesses (Sharma, and Chrisman, 1999; Verbeke et al., 2007). Although there are many studies focusing on one of these aspects (cf. Burgelman, 1985; Huff et al., 1992), it is very difficult to compare the outcomes of these studies due to differences in contexts, measures, and methodologies. This is a significant lack in our understanding of CE, as firms engage in all three aspects of CE, making it important to compare managerial and organizational antecedents of innovation, venturing and renewal processes. Because factors that may enhance venturing activities may simultaneously inhibit renewal activities. The limited studies that did empirically compare the individual dimensions (cf. Zahra, 1996; Zahra et al., 2000; Yiu and Lau, 2008; Yiu et al., 2007) generally failed to provide a systematic explanation for potential different effects of organizational antecedents on innovation, venturing, and renewal. With this study we aim to investigate the different effects of structural separation and social capital on corporate entrepreneurship activities. The access to existing and the development of new knowledge has been deemed of critical importance in CE-activities (Floyd and Wooldridge, 1999; Covin and Miles, 2007; Katila and Ahuja, 2002). Developing new knowledge can be facilitated by structurally separating corporate entrepreneurial units from mainstream units (cf. Burgelman, 1983; Hill and Rothaermel, 2003; O'Reilly and Tushman, 2004). Existing knowledge and resources are available through networks of social relationships, defined as social capital (Nahapiet and Ghoshal, 1998; Yiu and Lau, 2008). Although social capital has primarily been studied at the organizational level, it might be equally important at top management level (Belliveau et al., 1996). However, little is known about the joint effects of structural separation and integrative mechanisms to provide access to social capital on corporate entrepreneurship. Could these integrative mechanisms for example connect the separated units to facilitate both knowledge creation and sharing? Do these effects differ for innovation, venturing, and renewal processes? Are the effects different for organizational versus top management team integration mechanisms? Corporate entrepreneurship activities have for example been suggested to take place at different levels. Whereas innovation is suggested to be a more bottom-up process, strategic renewal is a more top-down process (Floyd and Lane, 2000; Volberda et al., 2001). Corporate venturing is also a more bottom-up process, but due to the greater required resource commitments relative to innovation, it ventures need to be approved by top management (Burgelman, 1983). As such we will explore the following key research question in this paper: How do social capital and structural separation on organizational and TMT level differentially influence innovation, venturing, and renewal processes? Methodology/Key Propositions We investigated our hypotheses on a final sample of 240 companies in a variety of industries in the Netherlands. All our measures were validated in previous studies. We targeted a second respondent in each firm to reduce problems with single-rater data (James et al., 1984). We separated the measurement of the independent and the dependent variables in two surveys to create a one-year time lag and reduce potential common method bias (Podsakoff et al., 2003). Results and Implications Consistent with our hypotheses, our results show that configurations of structural separation and integrative mechanisms have different effects on the three aspects of corporate entrepreneurship. Innovation was affected by organizational level mechanisms, renewal by integrative mechanisms on top management team level and venturing by mechanisms on both levels. Surprisingly, our results indicated that integrative mechanisms on top management team level had negative effects on corporate entrepreneurship activities. We believe this paper makes two significant contributions. First, we provide more insight in what the effects of ambidextrous organizational forms (i.e. combinations of differentiation and integration mechanisms) are on venturing, innovation and renewal processes. Our findings show that more valuable insights can be gained by comparing the individual parts of corporate entrepreneurship instead of focusing on the whole. Second, we deliver insights in how management can create a facilitative organizational context for these corporate entrepreneurship activities.

Business Creation Processes in Australia : What Start-Up Attempts Get Up and Running, and Why? - A Preliminary Assessment

The Comprehensive Australian Study of Entrepreneurial Emergence (CAUSEE) is the largest study of new firm formation that has ever been undertaken in Australia. CAUSEE follows the development of several samples of new and emerging firms over time. In this report we focus on the drivers of outcomes – in terms of reaching an operational stage vs. terminating the effort – of 493 randomly selected nascent firms whose founders have been comprehensively interviewed on two occasions, 12 months apart. We investigate the outcome effects of three groups of variables: Characteristics of the Venture; Resources Used in the Start-Up Process and Characteristics of the Start-Up Process Itself.

Primary students' group metacognitive processes in a computer supported collaborative learning environment

The current understanding of students’ group metacognition is limited. The research on metacognition has focused mainly on the individual student. The aim of this study was to address the void by developing a conceptual model to inform the use of scaffolds to facilitate group metacognition during mathematical problem solving in computer supported collaborative learning (CSCL) environments. An initial conceptual framework based on the literature from metacognition, cooperative learning, cooperative group metacognition, and computer supported collaborative learning was used to inform the study. In order to achieve the study aim, a design research methodology incorporating two cycles was used. The first cycle focused on the within-group metacognition for sixteen groups of primary school students working together around the computer; the second cycle included between-group metacognition for six groups of primary school students working together on the Knowledge Forum® CSCL environment. The study found that providing groups with group metacognitive scaffolds resulted in groups planning, monitoring, and evaluating the task and team aspects of their group work. The metacognitive scaffolds allowed students to focus on how their group was completing the problem-solving task and working together as a team. From these findings, a revised conceptual model to inform the use of scaffolds to facilitate group metacognition during mathematical problem solving in computer supported collaborative learning (CSCL) environments was generated.

Object-centric process models and the design of flexible processes

Mainstream business process modelling techniques promote a design paradigm wherein the activities to be performed within a case, together with their usual execution order, form the backbone of a process model, on top of which other aspects are anchored. This paradigm, while eective in standardised and production-oriented domains, shows some limitations when confronted with processes where case-by-case variations and exceptions are the norm. In this thesis we develop the idea that the eective design of exible process models calls for an alternative modelling paradigm, one in which process models are modularised along key business objects, rather than along activity decompositions. The research follows a design science method, starting from the formulation of a research problem expressed in terms of requirements, and culminating in a set of artifacts that have been devised to satisfy these requirements. The main contributions of the thesis are: (i) a meta-model for object-centric process modelling incorporating constructs for capturing exible processes; (ii) a transformation from this meta-model to an existing activity-centric process modelling language, namely YAWL, showing the relation between object-centric and activity-centric process modelling approaches; and (iii) a Coloured Petri Net that captures the semantics of the proposed meta-model. The meta-model has been evaluated using a framework consisting of a set of work ow patterns. Moreover, the meta-model has been embodied in a modelling tool that has been used to capture two industrial scenarios.

In-session processes in online counselling with young people : an exploratory approach

Aims : The aim of this study was to conduct an exploratory investigation into the in-session processes and behaviours that occur between therapists and young people in online counseling. Method: The Consensual Qualitative Research method was employed to identify in-session behaviours and a coding instrument was developed to determine their frequency of use and assess whether nuances carried in the meaning of text messages have an influential effect during sessions. Eighty-five single-session transcripts were examined in total by two independent coders. Results: Sample statistics revealed that, on average, rapport-building processes were used more consistently across cases with both types of processes having a moderately strong positive effect on young people. However, closer examination of these processes revealed weaker positive effects for in-session behaviours that rely more heavily on verbal and non-verbal cues to be accurately interpreted. Implications for Practice and Future Research: These findings imply that therapists may focus more on building rapport than accomplishing tasks with young people during online counselling sessions due to the absence of verbal and non-verbal information when communicating via text messages.

Framework and processes for enhancing sustainability deliverables in Australian road infrastructure projects

Road and highway infrastructure provides the backbone for a nation’s economic growth. The versatile dispersion of population in Australia and its resource boom, coupled with improved living standards and growing societal expectations, calls for continuing development and improvement of road infrastructure under the current local, state and federal governments’ policies and strategic plans. As road infrastructure projects involve huge resources and mechanisms, achieving sustainability not only on economic scales but also through environmental and social responsibility becomes a crucial issue. While sustainability is a logical link to infrastructure development, literature study and consultation with the industry found that there is a lack of common understanding on what constitutes sustainability in the infrastructure context. Its priorities are often interpreted differently among multiple stakeholders. For road infrastructure projects which typically span over long periods of time, achieving tangible sustainability outcomes during the lifecycle of development remains a formidable task. Sustainable development initiatives often remain ideological as in macro-level policies and broad-based concepts. There were little elaboration and exemplar cases on how these policies and concepts can be translated into practical decision-making during project implementation. In contrast, there seemed to be over commitment on research and development of sustainability assessment methods and tools. Between the two positions, there is a perception-reality gap and mismatch, specifically on how to enhance sustainability deliverables during infrastructure project delivery. Review on past research in this industry sector also found that little has been done to promote sustainable road infrastructure development; this has wide and varied potential impacts. This research identified the common perceptions and expectations by different stakeholders towards achieving sustainability in road and highway infrastructure projects. Face to face interviews on selected representatives of these stakeholders were carried out in order to select and categorize, confirm and prioritize a list of sustainability performance targets identified through literature and past research. A Delphi study was conducted with the assistance of a panel of senior industry professionals and academic experts, which further considered the interrelationship and influence of the sustainability indicators, and identified critical sustainability indicators under ten critical sustainability criteria (e.g. Environmental, Health & Safety, Resource Utilization & Management, Social & Cultural, Economic, Public Governance & Community Engagement, Relations Management, Engineering, Institutional and Project Management). This presented critical sustainability issues that needed to be addressed at the project level. Accordingly, exemplar highway development projects were used as case studies to elicit solutions for the critical issues. Through the identification and integration of different perceptions and priority needs of the stakeholders, as well as key sustainability indicators and solutions for critical issues, a set of decision-making guidelines was developed to promote and drive consistent sustainability deliverables in road infrastructure projects.

A tale of two cities: Education responds to globalisation in Hong Kong and Singapore in the aftermath of the Asian Economic Crisis

It has been suggested that although the most theorisation about globalisation has emerged from “western” contexts, the material implications of globalisation have been felt most strongly in non-western regions. With this in mind, we are undertaking a situated analysis of how two states, Singapore and Hong Kong, are interacting with the broader processes of globalisation through their educational policies. We apply Foucault's conceptual tool of governmentality to understand (i) the conduct of governing in the contemporary nation-state, and (ii) how the “right” rationalities are being inculcated by government to create “desiring subjects” who will play their part in ensuring national prosperity. We use the Asian Economic Crisis as a point of departure to show how global-local tensions are being managed by Singapore and Hong Kong. We conclude that both these global cities have adroitly managed the Asian economic crisis to steer their citizens away from pursuits of greater political freedom and towards concerns of material well being. They have done so through a selective interpretation of globalisation, by simultaneously resisting and embracing the contradictory strands of globalisation. Education has emerged as a critical space for this selective absorption of globalising trends.

Phenyl-ring disorder in the two-dimensional hydrogen-bonded structure of the 1:1 proton-transfer salt of the diazo-dye precursor 4-(phenyldiazenyl)aniline (aniline yellow) with L-tartaric acid

The structure of the 1:1 proton-transfer compound from the reaction of L-tartaric acid with the azo-dye precursor aniline yellow [4-(phenylazo)aniline], 4-(phenyldiazenyl)anilinium hydrogen 2R,3R-tartrate C12H12N3+ . C4H6O6- has been determined at 200 K. The asymmetric unit of the compound contains two independent phenylazoanilinium cations and two hydrogen L-tartrate anions. The structure is unusual in that all four phenyl rings of both cations have identical 50% rotational disorder. The two hydrogen L-tartrate anions form independent but similar chains through head-to-tail carboxylic O--H...O~carboxyl~ hydrogen bonds [graph set C7] which are then extended into a two-dimensional hydrogen-bonded sheet structure through hydroxyl O--H...O hydrogen-bonding links. The anilinium groups of the phenyldiazenyl cations are incorporated into the sheets and also provide internal hydrogen-bonding extensions while their aromatic tails layer in the structure without significant interaction except for weak \p--\p interactions [minimum ring centroid separation, 3.844(3) \%A]. The hydrogen L-tartrate residues of both anions have the common short intramolecular hydroxyl O--H...O~carboxyl~ hydogen bonds. This work has provided a solution to the unusual disorder problem inherent in the structure of this salt as well as giving another example of the utility of the hydrogen tartrate in the generation of sheet substructures in molecular assembly processes.