The synthesis of novel nitroxides and their application as small-molecule antioxidants and profluorescent probes for oxidative stress

The detection and potential treatment of oxidative stress in biological systems has been explored using isoindoline-based nitroxide radicals. A novel tetraethyl-fluorescein nitroxide was synthesised for its use as a profluorescent probe for redox processes in biological systems. This tetraethyl system, as well as a tetramethyl-fluorescein nitroxide, were shown to be sensitive and selective probes for superoxide in vitro. The redox environment of cellular systems was also explored using the tetramethylfluorescein species based on its reduction to the hydroxylamine. Flow cytometry was employed to assess the extent of nitroxide reduction, reflecting the overall cellular redox environment. Treatment of normal fibroblasts with rotenone and 2-deoxyglucose resulted in an oxidising cellular environment as shown by the lack of reduction of the fluorescein-nitroxide system. Assessment of the tetraethyl-fluorescein nitroxide system in the same way demonstrated its enhanced resistance to reduction and offers the potential to detect and image biologically relevant reactive oxygen species directly. Importantly, these profluorescent nitroxide compounds were shown to be more effective than the more widely used and commercially available probes for reactive oxygen species such as 2’,7’-dichlorodihydrofluorescein diacetate. Fluorescence imaging of the tetramethyl-fluorescein nitroxide and a number of other rhodamine-nitroxide derivatives was undertaken, revealing the differential cellular localisation of these systems and thus their potential for the detection of redox changes in specific cellular compartments. As well as developing novel methods for the detection of oxidative stress, a number of novel isoindoline nitroxides were synthesised for their potential application as small-molecule antioxidants. These compounds incorporated known pharmacophores into the isoindoline-nitroxide structure in an attempt to increase their efficacy in biological systems. A primary and a secondary amine nitroxide were synthesised which incorporated the phenethylamine backbone of the sympathomimetic amine class of drugs. Initial assessment of the novel primary amine derivative indicated a protective effect comparable to that of 5-carboxy-1,1,3,3- tetramethylisoindolin-2-yloxyl. Methoxy-substituted nitroxides were also synthesised as potential antioxidants for their structural similarity to some amphetamine type stimulants. A copper-catalysed methodology provided access to both the mono- and di-substituted methoxy-nitroxides. Deprotection of the ethers in these compounds using boron tribromide successfully produced a phenolnitroxide, however the catechol moiety in the disubstituted derivative appeared to undergo reaction with the nitroxide to produce quinone-like degradation products. A novel fluoran-nitroxide was also synthesised from the methoxy-substituted nitroxide, providing a pH-sensitive spin probe. An amino-acid precursor containing a nitroxide moiety was also synthesised for its application as a dual-action antioxidant. N-Acetyl protection of the nitroxide radical was necessary prior to the Erlenmeyer reaction with N-acetyl glycine. Hydrolysis and reduction of the azlactone intermediate produced a novel amino acid precursor with significant potential as an effective antioxidant.

Solving navigational uncertainty using grid cells on robots

To successfully navigate their habitats, many mammals use a combination of two mechanisms, path integration and calibration using landmarks, which together enable them to estimate their location and orientation, or pose. In large natural environments, both these mechanisms are characterized by uncertainty: the path integration process is subject to the accumulation of error, while landmark calibration is limited by perceptual ambiguity. It remains unclear how animals form coherent spatial representations in the presence of such uncertainty. Navigation research using robots has determined that uncertainty can be effectively addressed by maintaining multiple probabilistic estimates of a robot's pose. Here we show how conjunctive grid cells in dorsocaudal medial entorhinal cortex (dMEC) may maintain multiple estimates of pose using a brain-based robot navigation system known as RatSLAM. Based both on rodent spatially-responsive cells and functional engineering principles, the cells at the core of the RatSLAM computational model have similar characteristics to rodent grid cells, which we demonstrate by replicating the seminal Moser experiments. We apply the RatSLAM model to a new experimental paradigm designed to examine the responses of a robot or animal in the presence of perceptual ambiguity. Our computational approach enables us to observe short-term population coding of multiple location hypotheses, a phenomenon which would not be easily observable in rodent recordings. We present behavioral and neural evidence demonstrating that the conjunctive grid cells maintain and propagate multiple estimates of pose, enabling the correct pose estimate to be resolved over time even without uniquely identifying cues. While recent research has focused on the grid-like firing characteristics, accuracy and representational capacity of grid cells, our results identify a possible critical and unique role for conjunctive grid cells in filtering sensory uncertainty. We anticipate our study to be a starting point for animal experiments that test navigation in perceptually ambiguous environments.

How HCI design influences web security decisions

Even though security protocols are designed to make computer communication secure, it is widely known that there is potential for security breakdowns at the human machine interface. This paper reports on a diary study conducted in order to investigate what people identify as security decisions that they make while using the web. The study aimed to uncover how security is perceived in the individual's context of use. From this data, themes were drawn, with a focus on addressing security goals such as confidentiality and authentication. This study is the first study investigating users' web usage focusing on their self-documented perceptions of security and the security choices they made in their own environment.

Characterisation of the tritrophic interactions between tobacco yellow dwarf virus, its vector and host-plants

Tobacco yellow dwarf virus (TbYDV, family Geminiviridae, genus Mastrevirus) is an economically important pathogen causing summer death and yellow dwarf disease in bean (Phaseolus vulgaris L.) and tobacco (Nicotiana tabacum L.), respectively. Prior to the commencement of this project, little was known about the epidemiology of TbYDV, its vector and host-plant range. As a result, disease control strategies have been restricted to regular poorly timed insecticide applications which are largely ineffective, environmentally hazardous and expensive. In an effort to address this problem, this PhD project was carried out in order to better understand the epidemiology of TbYDV, to identify its host-plant and vectors as well as to characterise the population dynamics and feeding physiology of the main insect vector and other possible vectors. The host-plants and possible leafhopper vectors of TbYDV were assessed over three consecutive growing seasons at seven field sites in the Ovens Valley, Northeastern Victoria, in commercial tobacco and bean growing properties. Leafhoppers and plants were collected and tested for the presence of TbYDV by PCR. Using sweep nets, twenty-three leafhopper species were identified at the seven sites with Orosius orientalis the predominant leafhopper. Of the 23 leafhopper species screened for TbYDV, only Orosius orientalis and Anzygina zealandica tested positive. Forty-two different plant species were also identified at the seven sites and tested. Of these, TbYDV was only detected in four dicotyledonous species, Amaranthus retroflexus, Phaseolus vulgaris, Nicotiana tabacum and Raphanus raphanistrum. Using a quadrat survey, the temporal distribution and diversity of vegetation at four of the field sites was monitored in order to assess the presence of, and changes in, potential host-plants for the leafhopper vector(s) and the virus. These surveys showed that plant composition and the climatic conditions at each site were the major influences on vector numbers, virus presence and the subsequent occurrence of tobacco yellow dwarf and bean summer death diseases. Forty-two plant species were identified from all sites and it was found that sites with the lowest incidence of disease had the highest proportion of monocotyledonous plants that are non hosts for both vector and the virus. In contrast, the sites with the highest disease incidence had more host-plant species for both vector and virus, and experienced higher temperatures and less rainfall. It is likely that these climatic conditions forced the leafhopper to move into the irrigated commercial tobacco and bean crop resulting in disease. In an attempt to understand leafhopper species diversity and abundance, in and around the field borders of commercially grown tobacco crops, leafhoppers were collected from four field sites using three different sampling techniques, namely pan trap, sticky trap and sweep net. Over 51000 leafhopper samples were collected, which comprised 57 species from 11 subfamilies and 19 tribes. Twentythree leafhopper species were recorded for the first time in Victoria in addition to several economically important pest species of crops other than tobacco and bean. The highest number and greatest diversity of leafhoppers were collected in yellow pan traps follow by sticky trap and sweep nets. Orosius orientalis was found to be the most abundant leafhopper collected from all sites with greatest numbers of this leafhopper also caught using the yellow pan trap. Using the three sampling methods mentioned above, the seasonal distribution and population dynamics of O. orientalis was studied at four field sites over three successive growing seasons. The population dynamics of the leafhopper was characterised by trimodal peaks of activity, occurring in the spring and summer months. Although O. orientalis was present in large numbers early in the growing season (September-October), TbYDV was only detected in these leafhoppers between late November and the end of January. The peak in the detection of TbYDV in O. orientalis correlated with the observation of disease symptoms in tobacco and bean and was also associated with warmer temperatures and lower rainfall. To understand the feeding requirements of Orosius orientalis and to enable screening of potential control agents, a chemically-defined artificial diet (designated PT-07) and feeding system was developed. This novel diet formulation allowed survival for O. orientalis for up to 46 days including complete development from first instar through to adulthood. The effect of three selected plant derived proteins, cowpea trypsin inhibitor (CpTi), Galanthus nivalis agglutinin (GNA) and wheat germ agglutinin (WGA), on leafhopper survival and development was assessed. Both GNA and WGA were shown to reduce leafhopper survival and development significantly when incorporated at a 0.1% (w/v) concentration. In contrast, CpTi at the same concentration did not exhibit significant antimetabolic properties. Based on these results, GNA and WGA are potentially useful antimetabolic agents for expression in genetically modified crops to improve the management of O. orientalis, TbYDV and the other pathogens it vectors. Finally, an electrical penetration graph (EPG) was used to study the feeding behaviour of O. orientalis to provide insights into TbYDV acquisition and transmission. Waveforms representing different feeding activity were acquired by EPG from adult O. orientalis feeding on two plant species, Phaseolus vulgaris and Nicotiana tabacum and a simple sucrose-based artificial diet. Five waveforms (designated O1-O5) were observed when O. orientalis fed on P. vulgaris, while only four (O1-O4) and three (O1-O3) waveforms were observed during feeding on N. tabacum and the artificial diet, respectively. The mean duration of each waveform and the waveform type differed markedly depending on the food source. This is the first detailed study on the tritrophic interactions between TbYDV, its leafhopper vector, O. orientalis, and host-plants. The results of this research have provided important fundamental information which can be used to develop more effective control strategies not only for O. orientalis, but also for TbYDV and other pathogens vectored by the leafhopper.

Do ontological deficiencies in modeling grammars matter?

Conceptual modeling grammars are a fundamental means for specifying information systems requirements. However, the actual usage of these grammars is only poorly understood. In particular, little is known about how properties of these grammars inform usage beliefs such as usefulness and ease of use. In this paper we use an ontological theory to describe conceptual modeling grammars in terms of their ontological deficiencies, and formulate two propositions in regard to how these ontological deficiencies influence primary usage beliefs. Using BPMN as an example modeling grammar, we surveyed 528 modeling practitioners to test the theorized relationships. Our results show that users of conceptual modeling grammars perceive ontological deficiencies to exist, and that these deficiency perceptions are negatively associated with usefulness and ease of use of these grammars. With our research we provide empirical evidence in support of the predictions of the ontological theory of modeling grammar expressiveness, and we identify previously unexplored links between conceptual modeling grammars and grammar usage beliefs. This work implies for practice a much closer coupling of the act of (re ) designing modeling grammars with usage-related success metrics.

Detection of dsRNA in Grapevines Showing Symptoms of Rupestris Stem Pitting Disease and the Variabilities Encountered

Rupestris stem pitting (rSP), a graft-transmissible grapevine disease, can be identified only by its reaction (pitted wood) on inoculated Vitis rupestris ‘St. George.’ DsRNA was extracted from grapevines from California and Canada that indexed positive for rSP on St. George. Two distinct dsRNA species (B and C) (Mr = 5.3 × 106 and 4.4 × 106, respectively) were detected from the stem tissue of rSP-positive samples. Although similar dsRNA species (B and C) were detected in extracts of grapevines from New York, the association of dsRNA B and C with rSP in New York samples was not consistent. Also, eight different dsRNAs, known to be associated with the powdery mildew fungus, Uncinula necator, were detected in leaves of New York samples. In New York, the dsRNAs were not observed in leaves or stem samples collected from June through late August during the 1988 and 1989 growing seasons, suggesting that dsRNA detection in the grape tissue is variable throughout the season. We suggest that dsRNA species B and C are associated with rSP disease. The inconsistent results with New York samples are discussed.

Sequence Changes in Six Variants of Rice Tungro Bacilliform Virus and Their Phylogenetic Relationships

The DNA of three biological variants, G1, Ic and G2, which originated from the same greenhouse isolate of rice tungro bacilliform virus (RTBV) at the International Rice Research Institute (IRRI), was cloned and sequenced. Comparison of the sequences revealed small differences in genome sizes. The variants were between 95 and 99% identical at the nucleotide and amino acid levels. Alignment of the three genome sequences with those of three published RTBV sequences (Phi-1, Phi-2 and Phi-3) revealed numerous nucleotide substitutions and some insertions and deletions. The published RTBV sequences originated from the same greenhouse isolate at IRRI 20, 11 and 9 years ago. All open reading frames (ORFs) and known functional domains were conserved across the six variants. The cysteine-rich region of ORF3 showed the greatest variation. When the six DNA sequences from IRRI were compared with that of an isolate from Malaysia (Serdang), similar changes were observed in the cysteine-rich region in addition to other nucleotide substitutions and deletions across the genome. The aligned nucleotide sequences of the IRRI variants and Serdang were used to analyse phylogenetic relationships by the bootstrapped parsimony, distance and maximum-likelihood methods. The isolates clustered in three groups: Serdang alone; Ic and G1; and Phi-1, Phi-2, Phi-3 and G2. The distribution of phylogenetically informative residues in the IRRI sequences shared with the Serdang sequence and the differing tree topologies for segments of the genome suggested that recombination, as well as substitutions and insertions or deletions, has played a role in the evolution of RTBV variants. The significance and implications of these evolutionary forces are discussed in comparison with badnaviruses and caulimoviruses.

Workplace Equality in International Organizations: A Way Forward

In this contribution, I am interested in how discrimination issues are manifested in employment relations in the United Nations (UN), a public forum to all states political leaders to advance their concerns, the World Bank, a financial organization that promotes economic development, mainly in developing countries, and the Consultative Group on International Agricultural Research (CGIAR), the eldest and largest global public program of the World Bank with a strategic network of diverse stakeholders that harnesses the best in science to produce more and better food, reduce poverty and sustain environments. Considering the immunity and privileges granted to international organizations, what are the current available legal procedures, at the national or international level, for workplace equality? How accountable and transparent are they, based on the practice of these organizations? Can discrimination biases that go beyond the known individual-based discrimination claims be identified? If so, how can they be challenged and changed? Based of the special position of international civil servants in international organizations and the duty to protect their fundamental rights, I claim that the limitation of opportunity by discriminatory biases and the psychic burden on the individual staff member, on daily basis, qualify for a workplace wrong and call for independent and impartial legal procedures that would ensure due process and fair treatment.

'Radiant with possibility': involving young people in creating a vision for the future of their community

This paper argues that young people need to be given the opportunity to recognise the interaction between their own understandings of the world as it is now and the vision of what it might become. To support this argument, we discuss an urban planning project, known as the Lower Mill Site Project, which involved active participation of high school students from the local community. The outcomes of this project demonstrate the positive contributions young people can make to the process of urban redevelopment, the advantages of using a participatory design approach, and the utopian possibilities that can emerge when young people are invited to be part of an intergenerational community project.

Environmental health risk assessment of dioxin exposure through foods in a Dioxin hot spot—Bien Hoa City, Vietnam

This study used the Australian Environmental Health Risk Assessment Framework to assess the human health risk of dioxin exposure through foods for local residents in two wards of Bien Hoa City, Vietnam. These wards are known hot-spots for dioxin and a range of stakeholders from central government to local levels were involved in this process. Publications on dioxin characteristics and toxicity were reviewed and dioxin concentrations in local soil, mud, foods, milk and blood samples were used as data for this risk assessment. A food frequency survey of 400 randomly selected households in these wards was conducted to provide data for exposure assessment. Results showed that local residents who had consumed locally cultivated foods, especially fresh water fish and bottom-feeding fish, free-ranging chicken, duck, and beef were at a very high risk, with their daily dioxin intake far exceeding the tolerable daily intake recommended by the WHO. Based on the results of this assessment, a multifaceted risk management program was developed and has been recognized as the first public health program ever to have been implemented in Vietnam to reduce the risks of dioxin exposure at dioxin hot-spots.

Eschewing the fat: strong production but Neil LaBute below his best : a review

AWARD-WINNING American play and screen writer Neil LaBute is known for producing character-driven dramas that concentrate on the darker side of human nature and desire. In Fat Pig, LaBute picks up on a familiar theme: the way a perverse social preference for physical perfection affects human relationships. It is a topic LaBute has tackled before in The Shape of Things, a compelling play in which a beautiful young woman's efforts to help her new boyfriend pursue a program of self-improvement are eventually revealed to be part of a bizarre human experiment for her master-of-fine-arts degree.

Left vs right in black and white : a review

LIKE much of the work that David Williamson is known for, Let the Sunshine concentrates on tensions between characters who operate mainly as mouthpieces for opposing ideologies. Left-wing documentary-maker Toby and his wife Ros have moved to Noosa to escape the rat race in Sydney and some bad press surrounding one of Toby's projects. Trying to make social connections in town, Ros has reconnected with high school classmate Natasha, now the cosmetically-enhanced wife of wealthy right-wing property developer Ron. The posturing and conflict between Toby and Ron come to a head when the women invite their grown children -- struggling songwriter Rick and stressed corporate lawyer Emma -- to dinner to celebrate Toby's birthday, and the results of this encounter drive the rest of the plot. The scenario of Let the Sunshine is contrived, the characters are stereotyped, and their conflicts are little more than an old clash of ideologies cast loosely across the mainstream news media's characterisation of the sides in debates about development, climate change and the economic crisis.

A lesson in tension yet to be taut : a review

NOBEL prize-winning British writer Harold Pinter is best known for his social commentaries and for dark, sometimes violent "comedies of menace" that capture the cruelty at the heart of human relationships. It has been a while since a Pinter play was on the Brisbane stage, and it is exciting to see one of his lesser known early works, Betrayal, as part of Queensland Theatre Company's session. Betrayal moves backwards from a potentially scandalous moment. In a pub one night, Emma tells Jerry that his best friend Robert is divorcing her. Robert has been unfaithful. And, what's more, Robert now knows about their lengthy affair. What follows is a series of scenes that look back at the stages of that affair and the secrets that have lain unspoken at the heart of their comfortable middle-class lives for so long. Betrayal is characterised by the sparsity of action and language, dark humour, and sudden outbreaks of seemingly excessive cruelty that are a trademark of Pinter's writing for the theatre.

Dark humour invigorates familiar fables ; a review

THEATRE: Grimm Tales. By Carol Ann Duffy and Tim Supple. Queensland Theatre Company, Brisbane. November 16. QUEENSLAND Theatre Company concludes its season with Grimm Tales, Carol Ann Duffy and Tim Supple's adaptation of classic cautionary tales as set down by the Brothers Grimm in the 19th century. This programming decision is clearly designed to present fun family entertainment as Christmas approaches. In Grimm Tales, well-known stories such as Hansel and Gretel, Snow White and Rumpelstiltskin pack a little more punch than in your standard picture book. Duffy and Supple's play is by no means the sort of poetic, postmodern or politicised adaptation of the fairytale we see from writers such as Angela Carter, and it is not intended to be subversive or to question the social and gender assumptions that underpin the tales. Start of sidebar. Skip to end of sidebar. End of sidebar. Return to start of sidebar. Rather, a return to the grislier original incarnations of the tales - the wicked stepsisters who lop off parts of their feet to fit the slipper and win the prince, or the hare so confused by the hedgehog's stratagem to make him think he is losing the race, he runs himself to death - has a comic effect. In this production, directed by Michael Futcher, heightened performances from some of Brisbane's best comic and physical actors, live music and an open, acknowledged relationship with the audience establish the atmosphere for the piece. While the production is a little sombre and slow to start with the first tale, Hansel and Gretel, the knowingness and almost slapstick quality with which the cast plays out the gruesome, scatological or silly moments in the other tales are well pitched to carry the comedy. The action is supported by a fantastic set by Greg Clarke of wooden planked walls, stairs and walkways which, with the help of David Walter's lighting design, is transformed into forests, ballrooms and castles as the cast moves up, over and under it. The overall highlight is probably the cast Futcher has brought together. Established QTC actors Eugene Gilfedder, Lucas Stibbard and Scott Witt, and emerging QTC actor Melanie Zanetti, join Liz Buchanan, Dan Crestani and Emma Pursey, all well known for their independent work in Brisbane but making their mainstage debut for the QTC. Every one of them metamorphoses with ease from character to character, human to animal, and central player to support. There is nothing particularly new in Grimm Tales, and it doesn't try to do anything more (or, indeed, less) than entertain, but skilful direction and a strong cast ensure it succeeds on those terms.

Contemporary take on deconstructed Medea : a review

THEATRE: The New Dead: Medea Material. By Heiner Muller. Stella Electrika in association with La Boite Theatre Company, Brisbane, November 19. THERE has been a lot of intensity in independent theatre in Brisbane during the past year, as companies, production houses and producers have begun building new programs and platforms to support an expansion of pathways within the local theatre ecology. Audiences have been exposed to works signalling the diversity of what Brisbane theatre makers want to see on stage, from productions of new local and international pieces to new devised works, and the results of residencies and development programs. La Boite Theatre Company closes its inaugural indie season with a work that places it at the contemporary, experimental end of the spectrum. The New Dead: Medea Material is emerging director Kat Henry's interpretation of Heiner Muller's 1981 text Despoiled Shore Medea Material Landscape with Argonauts. Start of sidebar. Skip to end of sidebar. End of sidebar. Return to start of sidebar. Muller is known for his radical adaptations of historical dramas, from the Greeks to Shakespeare, and for deconstructed texts in which the characters - in this case, Medea - violently reject the familial, cultural and political roles society has laid out for them. Muller's combination of deconstructed characters, disconnected poetic language and constant references to aspects of popular culture and the Cold War politics he sought to abjure make his texts challenging to realise. The poetry entices but the density, together with the increasing distance of the Cold War politics in the texts, leaves contemporary directors with clear decisions to make about how to adapt these open texts. In The New Dead: Medea Material, Henry works with some interesting imagery and conceptual territory. Lucinda Shaw as Medea, Guy Webster as Jason and Kimie Tsukakoshi as King Creon's daughter Glauce, the woman for whom Jason forsakes his wife Medea, each reference different aspects of contemporary culture. Medea is a bitter, drunken, satin-gowned diva with bite; Jason - first seen lounging in front of the television with a beer in an image reminiscent of Sarah Kane's in-yer-face characterisation of Hippolytus in Phaedra's Love - has something of the rock star about him; and Glauce is a roller-skating, karaoke-singing, pole-dancing young temptress. The production is given a contemporary tone, dominated by Medea's twisted love and loss, rather than by any commentary on her circumstances. Its strength is the aesthetic Henry creates, supported by live electro-pop music, a band stage that stands as a metaphor for Jason's sea voyage, and multimedia that inserts images of the story unfolding beyond these characters' speeches as sorts of subconscious flashes. While Tsukakoshi is engaging throughout, there are moments when Shaw and Webster's performances - particularly in the songs - are diminished by a lack of clarity. The result is a piece that, while slightly lacking in its realisation at times, undoubtedly flags Henry's facility as an emerging director and what she wants to bring to the Brisbane theatre scene.