The Wnt receptor Ryk is required for Wnt5a-mediated axon guidance on the contralateral side of the corpus callosum

Ryk (receptor related to tyrosine kinase) has been shown to be a novel Wnt receptor in both Caenorhabditis elegans and Drosophila melanogaster. Recently, Ryk-Wnt interactions were shown to guide corticospinal axons down the embryonic mouse spinal cord. Here we show that, in Ryk-deficient mice, cortical axons project aberrantly across the major forebrain commissure, the corpus callosum. Many mouse mutants have been described in which loss-of-function mutations result in the inability of callosal axons to cross the midline, thereby forming Probst bundles on the ipsilateral side. In contrast, loss of Ryk does not interfere with the ability of callosal axons to cross the midline but impedes their escape from the midline into the contralateral side. Therefore, Ryk(-/-) mice display a novel callosal guidance phenotype. We also show that Wnt5a acts as a chemorepulsive ligand for Ryk, driving callosal axons toward the contralateral hemisphere after crossing the midline. In addition, whereas callosal axons do cross the midline in Ryk(-/-) embryos, they are defasciculated on the ipsilateral side, indicating that Ryk also promotes fasciculation of axons before midline crossing. In summary, this study expands the emerging role for Wnts in axon guidance and identifies Ryk as a key guidance receptor in the establishment of the corpus callosum. Our analysis of Ryk function further advances our understanding of the molecular mechanisms underlying the formation of this important commissure.

Measurement of the aerodynamic performance of a scramjet-powered vehicle in a shock tunnel

A force balance system for measuring lift, thrust and pitching moment has been used to measure the performance of fueled scramjet-powered vehicle in the T4 Shock Tunnel at The University of Queensland. Detailed measurements have been made of the effects of different fuel flow rates corresponding to equivalence ratios between 0.0 and 1.5. For proposed scramjet-powered vehicles, the fore-body of the vehicle acts as part of the inlet to the engine and the aft-body acts as the thrust surface for the engine. This type of engine-integrated design leads to a strong coupling between the performance of the engine and the lift and trim characteristics of the vehicle. The measurements show that the lift force increased by approximately 50% and centre-of-pressure changed by approximately 10% of the chord of the vehicle when the equivalence ratio varied from 0.0 to 1.0. The results demonstrate the importance of engine performance to the overall aerodynamic characteristics of engine-integrated scramjet vehicles and that such characteristics can be measured in a shock tunnel.

The role of tissue transglutaminase (TG2) in regulating the tumour progression of the mouse colon carcinoma CT26

The multifunctional enzyme tissue transglutaminase (TG2) is reported to both mediate and inhibit tumour progression. To elucidate these different roles of TG2, we established a series of stable-transfected mouse colon carcinoma CT26 cells expressing a catalytically active (wild type) and a transamidating-inactive TG2 (Cys277Ser) mutant. Comparison of the TG2-transfected cells with the empty vector control indicated no differences in cell proliferation, apoptosis and susceptibility to doxorubicin, which correlated with no detectable changes in the activation of the transcription factor NF-?B. TG2-transfected cells showed increased expression of integrin ß3, and were more adherent and less migratory on fibronectin than control cells. Direct interaction of TG2 with ß3 integrins was demonstrated by immunoprecipitation, suggesting that TG2 acts as a coreceptor for fibronectin with ß3 integrins. All cells expressed the same level of TGFß receptors I and II, but only cells transfected with active TG2 had increased levels of TGFß1 and matrix-deposited fibronectin, which could be inhibited by TG2 site-directed inhibitors. Moreover, only cells transfected with active TG2 were capable of inhibiting tumour growth when compared to the empty vector controls. We conclude that in this colon carcinoma model increased levels of active TG2 are unfavourable to tumour growth due to their role in activation of TGFß1 and increased matrix deposition, which in turn favours increased cell adhesion and a lowered migratory and invasive behaviour.

Inward investment and host country market structure:the case of the U.K.

Multinational enterprises are seen as vehicles for the international transfer of investment capital, protecting and increasing profits by transferring ownership advantages across national boundaries. As such, the argument often follows that foreign direct investment then exacerbates the monopoly problem in host countries, by increasing concentration and facilitating collusion. This paper however reveals the reverse, that inward investment into the U.K. acts to reduce concentration at the industry level, by increasing competitive pressures on domestic industry.

The effects of vigabatrin of the visual system

This thesis considers the visual electrophysiological effects of vigabatrin (an anti-epileptic drug, which acts by increasing the levels of the inhibitory neurotransmitter GABA on the retina of the eye compared to the concentric visual field defects which have been found associated with the drug. Flash and pattern ERG's, EOG's multifocal ERG's (VERIS), flash and pattern VEP's and visual fields were tested. Although VEP's have been shown not to be affected by vigabatrin, these were recorded to complete the testing. Initially, of the eight vigabatrin patients with known visual field defects, 7 showed abnormally delayed 30Hz flicker a-wave latencies, 5 abnormally delayed 30Hz b-wave latencies and 6 abnormally low 30Hz amplitudes. Also 7 showed an abnormally prolonged latency of oscillatory potential 1 (OP1). The two patients taking vigabatrin at the time of testing showed low EOG Arden index values. The VERIS results correlated well with the severity of the visual field defects. Following this finding, eleven healthy subjects received vigabatrin over a 10-day period. No changes were seen in the visual fields, however, the photopic ERG b-wave latency significantly increased (although not to abnormal values). A matched pairs study with eleven vigabatrin, patients and eleven epileptic patients, who had never taken vigabatrin supported the findings of abnormal 30Hz flicker b-wave and OP latencies associated with vigabatrin, again with the VERIS results correlating to the severity of the visual field defect. The abnormal 30Hz flicker and VERIS responses indicate involvement of the cone photoreceptors and the OP's show an effect on the amacrine cells. The ERG increase in the photopic b-wave latency also suggests involvement of the bipolar cells, however, this effect and the reversible effect on the Arden index after cessation of the drug may be unrelated to the visual field defect. To conclude this thesis, a field specific VEP stimulus was developed to assess the retinal function in the peripheral field of paediatric patients. It comprises of a dartboard stimulus with a central 0-5 degree black and white chequered stimulus, a blank 5-30 degree annulus and a 30-60 degree peripheral chequered stimulus. When optimised on four vigabatrin patients it was found that no peripheral response can be evoked with a field loss exceeding 30-35 degrees. Co-operation was found to be successful in children as young as four years old.

An insight into the activation mechanism of the calcatonin-gene-related peptide receptor

The calcitonin-gene- related peptide (CGRP) receptor is unique among G-protein coupled receptors (GPCRs) as it consists of at least three proteins: calcitonin receptor like receptor (CLR), receptor activity modifying protein (RAMP)1 and receptor component protein (RCP). An endogenous agonist for this curious receptor is aCGRP, which is a sensory nerve-derived peptide made up of 37 amino acids. aCGRP acts as a potent vasodilator having pronounced effects on arterioles and capillaries. Understanding the pharmacodynamics of the CGRP receptor may have pharmaceutical benefit as the receptor has been associated with the onset of migraines and implicated in Raynauds syndrome. The primary aim of this thesis was to identify functionally important residues in the extracellular face of the CGRP receptor. Three areas of interest were selected including the extreme N-terminus of the CLR, extracellular loop 1 (ECL1) of the CLR and its associated transmembrane (TM) regions, and finally extracellular loop 3 (ECL3) of the CLR and its juxtamembrane regions. A site-directed mutagenesis (SDM) strategy was used to investigate these regions, primarily substituting the innate residues of CLR with alanine and assessing the mutation on multiple criteria including a functional cAMP assay, cell-surface expression, total expression, agonist-mediated internalisation and aCGRP binding. The results are interpreted and discussed taking into consideration contemporary concepts surrounding Secretin-like GPCRs. Moreover, the thesis also contains details of RAMP purification. Overall the thesis provides novel data that furthers insight into the complex phenomenon of CGRP receptor activation. Site-directed mutants have been identified that affect aCGRP binding, receptor signal transduction, the CLR/RAMP1 interface and the integrity of the protein complex structure.

Mapping interaction sites within the N-terminus of the calcitonin gene-related peptide receptor; the role of residues 23-60 of the calcitonin receptor-like receptor

The calcitonin receptor-like receptor (CLR) acts as a receptor for the calcitonin gene-related peptide (CGRP) but in order to recognize CGRP, it must form a complex with an accessory protein, receptor activity modifying protein 1 (RAMP1). Identifying the protein/protein and protein/ligand interfaces in this unusual complex would aid drug design. The role of the extreme N-terminus of CLR (Glu23-Ala60) was examined by an alanine scan and the results were interpreted with the help of a molecular model. The potency of CGRP at stimulating cAMP production was reduced at Leu41Ala, Gln45Ala, Cys48Ala and Tyr49Ala; furthermore, CGRP-induced receptor internalization at all of these receptors was also impaired. Ile32Ala, Gly35Ala and Thr37Ala all increased CGRP potency. CGRP specific binding was abolished at Leu41Ala, Ala44Leu, Cys48Ala and Tyr49Ala. There was significant impairment of cell surface expression of Gln45Ala, Cys48Ala and Tyr49Ala. Cys48 takes part in a highly conserved disulfide bond and is probably needed for correct folding of CLR. The model suggests that Gln45 and Tyr49 mediate their effects by interacting with RAMP1 whereas Leu41 and Ala44 are likely to be involved in binding CGRP. Ile32, Gly35 and Thr37 form a separate cluster of residues which modulate CGRP binding. The results from this study may be applicable to other family B GPCRs which can associate with RAMPs.

Cognitive systems associated with the hippocampus of the human brain and their role in behaviour and neurodegenerative disease

Cognitive systems research involves the synthesis of ideas from natural and artificial systems in the analysis, understanding, and design of all intelligent systems. This chapter discusses the cognitive systems associated with the hippocampus (HC) of the human brain and their possible role in behaviour and neurodegenerative disease. The hippocampus (HC) is concerned with the analysis of highly abstract data derived from all sensory systems but its specific role remains controversial. Hence, there have been three major theories concerning its function, viz., the memory theory, the spatial theory, and the behavioral inhibition theory. The memory theory has its origin in the surgical destruction of the HC, which results in severe anterograde and partial retrograde amnesia. The spatial theory has its origin in the observation that neurons in the HC of animals show activity related to their location within the environment. By contrast, the behavioral inhibition theory suggests that the HC acts as a ‘comparator’, i.e., it compares current sensory events with expected or predicted events. If a set of expectations continues to be verified then no alteration of behavior occurs. If, however, a ‘mismatch’ is detected then the HC intervenes by initiating appropriate action by active inhibition of current motor programs and initiation of new data gathering. Understanding the cognitive systems of the hippocampus in humans may aid in the design of intelligent systems involved in spatial mapping, memory, and decision making. In addition, this information may lead to a greater understanding of the course of clinical dementia in the various neurodegenerative diseases in which there is significant damage to the HC.

Residue-specific investigation of the relationship between oxidation and activity of a dual specificity phosphatase by mass spectrometry

Redox regulation of signalling pathways is critical in proliferation and apoptosis; redox imbalance can lead to pathologies such as inflammation and cancer. Vaccinia H1-related protein (VHR; DUSP3) is a dual-specificity phosphatase important in controlling MAP kinase activity during cell cycle. the active-site motif contains a cysteine that acts as a nucleophile during catalysis. We used VHR to investigate the effect of oxidation in vitro on phosphatase activity, with the aim of determining how the profile of site-specific modification related to catalytic activity. Recombinant human VHR was expressed in E. coli and purified using a GST-tag. Protein was subjected to oxidation with various concentrations of SIN-1 or tetranitromethane (TNM) as nitrating agents, or HOCl. the activity was assayed using either 3-O-methylfluorescein phosphate with fluorescence detection or PIP3 by phosphate release with malachite green. the sites of oxidation were mapped using HPLC coupled to tandem mass spectrometry on an ABSciex 5600TripleTOF following in-gel digestion. More than 25 different concentration-dependent oxidative modifications to the protein were detected, including oxidations of methionine, cysteine, histidine, lysine, proline and tyrosine, and the % oxidized peptide (versus unmodified peptide) was determined from the extracted ion chromatograms. Unsurprisingly, methionine residues were very susceptible to oxidation, but there was a significant different in the extent of their oxidation. Similarly, tyrosine residues varied greatly in their modifications: Y85 and Y138 were readily nitrated, whereas Y38, Y78 and Y101 showed little modification. Y138 must be phosphorylated for MAPK phosphatase activity, so this susceptibility impacts on signalling pathways. Di- and tri- oxidations of cysteine residues were observed, but did not correlate directly with loss of activity. Overall, the catalytic activity did not correlate with redox state of any individual residue, but the total oxidative load correlated with treatment concentration and activity. This study provides the first comprehensive analysis of oxidation modifications of VHR, and demonstrates both heterogenous oxidant effects and differential residue susceptibility in a signalling phosphatase.

A seismic shift? An evaluation of the impact of new media on perceptions of freedom of expression and privacy

New media platforms have changed the media landscape forever, as they have altered our perceptions of the limits of communication, and reception of information. Platforms such as Facebook, Twitter and WhatsApp enable individuals to circumvent the traditional mass media, converging audience and producer to create millions of ‘citizen journalists’. This new breed of journalist uses these platforms as a way of, not only receiving news, but of instantaneously, and often spontaneously, expressing opinions and venting and sharing emotions, thoughts and feelings. They are liberated from cultural and physical restraints, such as time, space and location, and they are not constrained by factors that impact upon the traditional media, such as editorial control, owner or political bias or the pressures of generating commercial revenue. A consequence of the way in which these platforms have become ingrained within our social culture is that habits, conventions and social norms, that were once informal and transitory manifestations of social life, are now infused within their use. What were casual and ephemeral actions and/or acts of expression, such as conversing with friends or colleagues or swapping/displaying pictures, or exchanging thoughts that were once kept private, or maybe shared with a select few, have now become formalised and potentially permanent, on view for the world to see. Incidentally, ‘traditional’ journalists and media outlets are also utilising new media, as it allows them to react, and disseminate news, instantaneously, within a hyper-competitive marketplace. However, in a world where we are saturated, not only by citizen journalists, but by traditional media outlets, offering access to news and opinion twenty-four hours a day, via multiple new media platforms, there is increased pressure to ‘break’ news fast and first. This paper will argue that new media, and the culture and environment it has created, for citizen journalists, traditional journalists and the media generally, has altered our perceptions of the limits and boundaries of freedom of expression dramatically, and that the corollary to this seismic shift is the impact on the notion of privacy and private life. Consequently, this paper will examine what a reasonable expectation of privacy may now mean, in a new media world.

Conformational analysis of the natural iron chelator myo-inositol 1,2,3-trisphosphate using a pyrene-based fluorescent mimic

myo-Inositol phosphates possessing the 1,2,3-trisphosphate motif share the remarkable ability to completely inhibit iron-catalysed hydroxyl radical formation. The simplest derivative, myo-inositol 1,2,3-trisphosphate [Ins(1,2,3)P3], has been proposed as an intracellular iron chelator involved in iron transport. The binding conformation of Ins(1,2,3)P3 is considered to be important to complex Fe3+ in a 'safe' manner. Here, a pyrene-based fluorescent probe, 4,6-bispyrenoyl-myo-inositol 1,2,3,5-tetrakisphosphate [4,6-bispyrenoyl Ins(1,2,3,5)P4], has been synthesised and used to monitor the conformation of the 1,2,3-trisphosphate motif using excimer fluorescence emission. Ring-flip of the cyclohexane chair to the penta-axial conformation occurs upon association with Fe3+, evident from excimer fluorescence induced by π-π stacking of the pyrene reporter groups, accompanied by excimer formation by excitation at 351 nm. This effect is unique amongst biologically relevant metal cations, except for Ca 2+ cations exceeding a 1:1 molar ratio. In addition, the thermodynamic constants for the interaction of the fluorescent probe with Fe3+ have been determined. The complexes formed between Fe 3+ and 4,6-bispyrenoyl Ins(1,2,3,5)P4 display similar stability to those formed with Ins(1,2,3)P3, indicating that the fluorescent probe acts as a good model for the 1,2,3-trisphosphate motif. This is further supported by the antioxidant properties of 4,6-bispyrenoyl Ins(1,2,3,5)P4, which closely resemble those obtained for Ins(1,2,3)P3. The data presented confirms that Fe3+ binds tightly to the unstable penta-axial conformation of myo-inositol phosphates possessing the 1,2,3-trisphosphate motif. © 2010 The Royal Society of Chemistry.

Measuring employee innovation:a review of existing scales and the development of the innovative behavior and innovation support inventories across cultures

Purpose - The paper develops a model of employee innovative behavior conceptualizing it as distinct from innovation outputs and as a multi-faceted behavior rather than a simple count of ‘innovative acts’ by employees. It understands individual employee innovative behaviors as a micro-foundation of firm intrapreneurship that is embedded in and influenced by contextual factors such as managerial, organizational and cultural support for innovation. Building from a review of existing employee innovative behavior scales and theoretical considerations we develop and validate the Innovative Behavior Inventory (IBI) and the Innovation Support Inventory (ISI). Design/methodology/approach – Two pilot studies, a third validation study in the Czech Republic and a fourth cross-cultural validation study using population representative samples from Switzerland, Germany, Italy and the Czech Republic (N=2812 employees and 450 entrepreneurs) were conducted. Findings - Both inventories were reliable and showed factorial, criterion, convergent and discriminant validity as well as cross-cultural equivalence. Employee innovative behavior was supported as comprising of idea generation, idea search, idea communication, implementation starting activities, involving others and overcoming obstacles. Managerial support was the most proximal contextual influence on innovative behavior and mediated the effect of organizational support and national culture. Originality/value - The paper advances our understanding of employee innovative behavior as a multi-faceted phenomenon and the contextual factors influencing it. Where past research typically focuses on convenience samples within a particular country, we offer first robust evidence that our model of employee innovative behavior generalizes across cultures and types of samples. Our model and the IBI and ISI inventories enable researchers to build a deeper understanding of the important micro-foundation underpinning intrapreneurial behavior in organizations and allow practitioners to identify their organizations’ strengths and weaknesses related to intrapreneurship.

Összegződnek-e az egyéni törekvések? A cselekvés és az eredmény közötti szakadék problémája (Can individual efforts be aggregated? The problem of the action-impact gap)

A szerző arra a kérdésre keresi a választ, hogyan alakulhat ki látványos szakadék az egyéni cselekvések iránya és azok együttes hatása között. A szándékok és tettek hatását felülírhatja a társadalmi gazdasági tényezőkből adódó tehetetlenség: a kritikus tömeg hiánya, szervezeti-infrastrukturális tényezők, kompenzációs hatások, egymás hatását kioltó cselekvések. A szerző a környezettudatosság és az ökológiai lábnyom példáján - ezerfős reprezentatív felmérésre alapozva - mutatja be, hogy az önkéntességre alapozó megközelítés sokszor túlbecsüli a fogyasztó - társadalmi-gazdasági tényezők által korlátozott - lehetőségeit és szuverenitását. _____ Behaviour impact gaps are demonstrably present in everyday life. It is increasingly found that environmental awareness in individuals fails to lead to reductions in the ecological footprint. Intensive agricultural practice reduces biodiversity in the EU even in areas where massive agri-environmental grant schemes are available and applied. Labour market training programmes do not necessarily facilitate job-finding for underprivileged segments of the society. So individual efforts may not add up or induce the expected effect. This outcome appears even for programmes that are successful in attaining the required behavioural change in a target group. The impact of attitudes and individual acts may be wiped out by structural and economic lock-ins such as trade-offs made for the gains, lack of a critical mass of actions, infrastructural deficiencies, or interfering acts of economic actors. The discrepancy between environmental awareness and ecological footprint is used to point out how awareness-raising programmes may miss their targets by overestimating the sovereignty and capabilities of consumers. Consumers are unwillingly locked into unsustainable practices and cannot be moved from that position unless economic and structural premises are also changed.

Analysis and comparison of the moral development of students required to graduate with an ethics course

Accounting students become practitioners facing ethical decision-making challenges that can be subject to various interpretations; hence, the profession is concerned with the appropriateness of their decisions. Moral development of these students has implications for a profession under legal challenges, negative publicity, and government scrutiny. Accounting students' moral development has been studied by examining their responses to moral questions in Rest's Defining Issues Test (DIT), their professional attitudes on Hall's Professionalism Scale Dimensions, and their ethical orientation-based professional commitment and ethical sensitivity. This study extended research in accounting ethics and moral development by examining students in a college where an ethics course is a requirement for graduation. ^ Knowledge of differences in the moral development of accounting students may alert practitioners and educators to potential problems resulting from a lack of ethical understanding as measured by moral development levels. If student moral development levels differ by major, and accounting majors have lower levels than other students, the conclusion may be that this difference is a causative factor for the alleged acts of malfeasance in the profession that may result in malpractice suits. ^ The current study compared 205 accounting, business, and nonbusiness students from a private university. In addition to academic major and completion of an ethics course, the other independent variable was academic level. Gender and age were tested as control variables and Rest's DIT score was the dependent variable. The primary analysis was a 2 x 3 x 3 ANOVA with post hoc tests for results with significant p-value of less than 0.05. ^ The results of this study reveal that students who take an ethics course appear to have a higher level of moral development (p = 0.013), as measured by the (DIT), than students at the same academic level who have not taken an ethics course. In addition, a statistically significant difference (p = 0.034) exists between freshmen who took an ethics class and juniors who did not take an ethics class. For every analysis except one, the lower class year with an ethics class had a higher level of moral development than the higher class year without an ethics class. These results appear to show that ethics education in particular has a greater effect on the level of moral development than education in general. Findings based on the gender specific analyses appear to show that males and females respond differently to the effects of taking an ethics class. The male students do not appear to increase their moral development level after taking an ethics course (p = 0.693) but male levels of moral development differ significantly (p = 0.003) by major. Female levels of moral development appear to increase after taking an ethics course (p = 0.002). However, they do not differ according to major (p = 0.097). ^ These findings indicate that accounting students should be required to have a class in ethics as part of their college curriculum. Students with an ethics class have a significantly higher level of moral development. The challenges facing the profession at the current time indicate that public confidence in the reports of client corporations has eroded and one way to restore this confidence could be to require ethics training of future accountants. ^