Ryanodine receptor phosphorylation in human heart failure

Heart failure is a complex disorder, characterized by activation of the sympathetic nervous system, leading to dysregulated Ca2+ homeostasis in cardiac myocytes and tissue remodeling. In a variety of diseases, cardiac malfunction is associated with aberrant fluxes of Ca2+ across both the surface membrane and the internal Ca2+ store, the sarcoplasmic reticulum (SR). One prominent hypothesis residues is that in heart failure, the activity of the ryanodine receptor (RyR2) Ca2+ release channel in the SR is increased due to excess phosphorylation and that this contributes to excess SR Ca2+ leak in diastole, reduced SR Ca2+ load and decreased contractility (Huke & Bers, 2008). There is controversy over which serine residues in RyR2 are hyperphosphorylated in animal models of heart failure and whether this is via the CaMKII or the PKA-linked signaling pathway. S2808, S2814 and S2030 in RyR2 have been variously claimed to be hyperphosphorylated. Our aim was to examine the degree of phosphorylation of these residues in RyR2 from failing human hearts. The use of human tissue was approved by the Human Research Ethics Committee, The Prince Charles Hospital, EC28114. Left ventricular tissue samples were obtained from an explanted heart of a patient with endstage heart failure (Emery Dreifuss Muscular Dystrophy with cardiomyopathy) and non-failing tissue was from a patient with cystic fibrosis undergoing heart-lung transplantation with no history of heart disease. SR vesicles were prepared as described by Laver et al. (1995) and examined with SDS-Page and Western Blot. Transferred proteins were probed with antibodies to detect total protein phosphorylation, phosphorylation of RyR2 serine residues S2808, S2814, S2030 and for the key proteins calsequestrin, triadin, junctin and FKBP12.6. To avoid membrane stripping artifact, each membrane was exposed to one phosphorylation-specific antibody and signal densities quantified using Bio-Rad Quantity One software. We found no distinguishable difference between failing and healthy hearts in the protein expression levels of RyR2, triadin, junctin or calsequestrin. We found an expected upregulation of total RyR2 phosphorylation in the failing heart sample, compared to a matched amount of RyR2 (quantified using densiometry) in healthy heart. Probing with antibodies detecting only the phosphorylated form of the specific RyR2 residues showed that the increase in total RyR2 phosphorylation in the failing heart was due to hyperphosphorylation of S2808 and S2814. We found that S2030 phosphorylation levels were unchanged in human heart failure. Interestingly, we found that S2030 has a basal level of phosphorylation in the healthy human heart, different from the absence of basal phosphorylation recently reported in rodent heart (Huke & Bers, 2008). Finally, preliminary results indicate that less FKBP 12.6 is associated with RyR2 in the failing heart, possibly as a consequence of PKA activation. In conclusion, residues S2808 and S2814 are hyperphosphorylated in human heart failure, presumably due to upregulation of the CaMKII and/or PKA signaling pathway as a result of chronic activation of the sympathetic nervous system. Such changes in RyR2 phosphorylation are believed to contribute to the leaky RyR2 phenotype associated with heart failure, which increases the incidence of arrhythmia and contributes to the severely impaired contractile performance of the failing heart. Huke S & Bers DM. (2008). Ryanodine receptor phosphorylation at serine 2030, 2808 and 2814 in rat cardiomyocytes. Biochemical and Biophysical Research Communications 376, 80-85. Laver DR, Roden LD, Ahern GP, Eager KR, Junankar PR & Dulhunty AF. (1995). Cytoplasmic Ca2+ inhibits the ryanodine receptor from cardiac muscle. Journal of Membrane Biology 147, 7-22. Proceedings

Using routinely collected hospital data for child maltreatment surveillance : issues, methods and patterns

Background: International data on child maltreatment are largely derived from child protection agencies, and predominantly report only substantiated cases of child maltreatment. This approach underestimates the incidence of maltreatment and makes inter-jurisdictional comparisons difficult. There has been a growing recognition of the importance of health professionals in identifying, documenting and reporting suspected child maltreatment. This study aimed to describe the issues around case identification using coded morbidity data, outline methods for selecting and grouping relevant codes, and illustrate patterns of maltreatment identified. Methods: A comprehensive review of the ICD-10-AM classification system was undertaken, including review of index terms, a free text search of tabular volumes, and a review of coding standards pertaining to child maltreatment coding. Identified codes were further categorised into maltreatment types including physical abuse, sexual abuse, emotional or psychological abuse, and neglect. Using these code groupings, one year of Australian hospitalisation data for children under 18 years of age was examined to quantify the proportion of patients identified and to explore the characteristics of cases assigned maltreatment-related codes. Results: Less than 0.5% of children hospitalised in Australia between 2005 and 2006 had a maltreatment code assigned, almost 4% of children with a principal diagnosis of a mental and behavioural disorder and over 1% of children with an injury or poisoning as the principal diagnosis had a maltreatment code assigned. The patterns of children assigned with definitive T74 codes varied by sex and age group. For males selected as having a maltreatment-related presentation, physical abuse was most commonly coded (62.6% of maltreatment cases) while for females selected as having a maltreatment-related presentation, sexual abuse was the most commonly assigned form of maltreatment (52.9% of maltreatment cases). Conclusion: This study has demonstrated that hospital data could provide valuable information for routine monitoring and surveillance of child maltreatment, even in the absence of population-based linked data sources. With national and international calls for a public health response to child maltreatment, better understanding of, investment in and utilisation of our core national routinely collected data sources will enhance the evidence-base needed to support an appropriate response to children at risk.

Supervisory safety practices in the work-related driving context

Work-related driving crashes are the most common cause of work-related injury, death, and absence from work in Australia and overseas. Surprisingly however, limited attention has been given to initiatives designed to improve safety outcomes in the work-related driving setting. This research paper will present preliminary findings from a research project designed to examine the effects of increasing work-related driving safety discussions on the relationship between drivers and their supervisors and motivations to drive safely. The research project was conducted within a community nursing population, where 112 drivers were matched with 23 supervisors. To establish discussions between supervisors and drivers, safety sessions were conducted on a monthly basis with supervisors of the drivers. At these sessions, the researcher presented context specific, audio-based anti-speeding messages. Throughout the course of the intervention and following each of these safety sessions, supervisors were instructed to ensure that all drivers within their workgroup listened to each particular anti-speeding message at least once a fortnight. In addition to the message, supervisors were also encouraged to frequently promote the anti-speeding message through any contact they had with their drivers (i.e., face to face, email, SMS text, and/or paper based contact). Fortnightly discussions were subsequently held with drivers, whereby the researchers ascertained the number and type of discussions supervisors engaged in with their drivers. These discussions also assessed drivers’ perceptions of the group safety climate. In addition to the fortnightly discussion, drivers completed a daily speed reporting form which assessed the proportion of their driving day spent knowingly over the speed limit. As predicted, the results found that if supervisors reported a good safety climate prior to the intervention, increasing the number of safety discussions resulted in drivers reporting a high quality relationship (i.e., leader-member exchange) with their supervisor post intervention. In addition, if drivers reported a good safety climate, increasing the number of discussions resulted in increased motivation to drive safely post intervention. Motivations to drive safely prior to the intervention also predicted self-reported speeding over the subsequent three months of reporting. These results suggest safety discussions play an important role in improving the exchange between supervisors and their drivers and drivers’ subsequent motivation to drive safely and, in turn, self reported speeding.

Clinical indicators of quality for Australian residential aged care facilities : establishing reliability, validity, and quality thresholds

Background: In response to the need for more comprehensive quality assessment within Australian residential aged care facilities, the Clinical Care Indicator (CCI) Tool was developed to collect outcome data as a means of making inferences about quality. A national trial of its effectiveness and a Brisbane-based trial of its use within the quality improvement context determined the CCI Tool represented a potentially valuable addition to the Australian aged care system. This document describes the next phase in the CCI Tool.s development; the aims of which were to establish validity and reliability of the CCI Tool, and to develop quality indicator thresholds (benchmarks) for use in Australia. The CCI Tool is now known as the ResCareQA (Residential Care Quality Assessment). Methods: The study aims were achieved through a combination of quantitative data analysis, and expert panel consultations using modified Delphi process. The expert panel consisted of experienced aged care clinicians, managers, and academics; they were initially consulted to determine face and content validity of the ResCareQA, and later to develop thresholds of quality. To analyse its psychometric properties, ResCareQA forms were completed for all residents (N=498) of nine aged care facilities throughout Queensland. Kappa statistics were used to assess inter-rater and test-retest reliability, and Cronbach.s alpha coefficient calculated to determine internal consistency. For concurrent validity, equivalent items on the ResCareQA and the Resident Classification Scales (RCS) were compared using Spearman.s rank order correlations, while discriminative validity was assessed using known-groups technique, comparing ResCareQA results between groups with differing care needs, as well as between male and female residents. Rank-ordered facility results for each clinical care indicator (CCI) were circulated to the panel; upper and lower thresholds for each CCI were nominated by panel members and refined through a Delphi process. These thresholds indicate excellent care at one extreme and questionable care at the other. Results: Minor modifications were made to the assessment, and it was renamed the ResCareQA. Agreement on its content was reached after two Delphi rounds; the final version contains 24 questions across four domains, enabling generation of 36 CCIs. Both test-retest and inter-rater reliability were sound with median kappa values of 0.74 (test-retest) and 0.91 (inter-rater); internal consistency was not as strong, with a Chronbach.s alpha of 0.46. Because the ResCareQA does not provide a single combined score, comparisons for concurrent validity were made with the RCS on an item by item basis, with most resultant correlations being quite low. Discriminative validity analyses, however, revealed highly significant differences in total number of CCIs between high care and low care groups (t199=10.77, p=0.000), while the differences between male and female residents were not significant (t414=0.56, p=0.58). Clinical outcomes varied both within and between facilities; agreed upper and lower thresholds were finalised after three Delphi rounds. Conclusions: The ResCareQA provides a comprehensive, easily administered means of monitoring quality in residential aged care facilities that can be reliably used on multiple occasions. The relatively modest internal consistency score was likely due to the multi-factorial nature of quality, and the absence of an aggregate result for the assessment. Measurement of concurrent validity proved difficult in the absence of a gold standard, but the sound discriminative validity results suggest that the ResCareQA has acceptable validity and could be confidently used as an indication of care quality within Australian residential aged care facilities. The thresholds, while preliminary due to small sample size, enable users to make judgements about quality within and between facilities. Thus it is recommended the ResCareQA be adopted for wider use.

Risk factor analysis and spatiotemporal CART model of cryptosporidiosis in Queensland, Australia

Background: It remains unclear whether it is possible to develop a spatiotemporal epidemic prediction model for cryptosporidiosis disease. This paper examined the impact of social economic and weather factors on cryptosporidiosis and explored the possibility of developing such a model using social economic and weather data in Queensland, Australia. ----- ----- Methods: Data on weather variables, notified cryptosporidiosis cases and social economic factors in Queensland were supplied by the Australian Bureau of Meteorology, Queensland Department of Health, and Australian Bureau of Statistics, respectively. Three-stage spatiotemporal classification and regression tree (CART) models were developed to examine the association between social economic and weather factors and monthly incidence of cryptosporidiosis in Queensland, Australia. The spatiotemporal CART model was used for predicting the outbreak of cryptosporidiosis in Queensland, Australia. ----- ----- Results: The results of the classification tree model (with incidence rates defined as binary presence/absence) showed that there was an 87% chance of an occurrence of cryptosporidiosis in a local government area (LGA) if the socio-economic index for the area (SEIFA) exceeded 1021, while the results of regression tree model (based on non-zero incidence rates) show when SEIFA was between 892 and 945, and temperature exceeded 32°C, the relative risk (RR) of cryptosporidiosis was 3.9 (mean morbidity: 390.6/100,000, standard deviation (SD): 310.5), compared to monthly average incidence of cryptosporidiosis. When SEIFA was less than 892 the RR of cryptosporidiosis was 4.3 (mean morbidity: 426.8/100,000, SD: 319.2). A prediction map for the cryptosporidiosis outbreak was made according to the outputs of spatiotemporal CART models. ----- ----- Conclusions: The results of this study suggest that spatiotemporal CART models based on social economic and weather variables can be used for predicting the outbreak of cryptosporidiosis in Queensland, Australia.

Physical and functional interaction of the archaeal single-stranded DNA-binding protein SSB with RNA polymerase

Archaeal transcription utilizes a complex multisubunit RNA polymerase and the basal transcription factors TBP and TF(II)B, closely resembling its eukaryal counterpart. We have uncovered a tight physical and functional interaction between RNA polymerase and the single-stranded DNA-binding protein SSB in Sulfolobus solfataricus. SSB stimulates transcription from promoters in vitro under TBP-limiting conditions and supports transcription in the absence of TBP. SSB also rescues transcription from repression by reconstituted chromatin. We demonstrate the potential for promoter melting by SSB, suggesting a plausible basis for the stimulation of transcription. This stimulation requires both the single-stranded DNA-binding domain and the acidic C-terminal tail of the SSB. The tail forms a stable interaction with RNA polymerase. These data reveal an unexpected role for single-stranded DNA-binding proteins in transcription in archaea.

Improving matching process in social network using implicit and explicit user information

Personalised social matching systems can be seen as recommender systems that recommend people to others in the social networks. However, with the rapid growth of users in social networks and the information that a social matching system requires about the users, recommender system techniques have become insufficiently adept at matching users in social networks. This paper presents a hybrid social matching system that takes advantage of both collaborative and content-based concepts of recommendation. The clustering technique is used to reduce the number of users that the matching system needs to consider and to overcome other problems from which social matching systems suffer, such as cold start problem due to the absence of implicit information about a new user. The proposed system has been evaluated on a dataset obtained from an online dating website. Empirical analysis shows that accuracy of the matching process is increased, using both user information (explicit data) and user behavior (implicit data).

Conformational flexibility revealed by the crystal structure of a crenarchaeal RadA

Homologous recombinational repair is an essential mechanism for repair of double-strand breaks in DNA. Recombinases of the RecA-fold family play a crucial role in this process, forming filaments that utilize ATP to mediate their interactions with singleand double-stranded DNA. The recombinase molecules present in the archaea (RadA) and eukaryota (Rad51) are more closely related to each other than to their bacterial counterpart (RecA) and, as a result, RadA makes a suitable model for the eukaryotic system. The crystal structure of Sulfolobus solfataricus RadA has been solved to a resolution of 3.2 A° in the absence of nucleotide analogues or DNA, revealing a narrow filamentous assembly with three molecules per helical turn. As observed in other RecA-family recombinases, each RadA molecule in the filament is linked to its neighbour via interactions of a short b-strand with the neighbouring ATPase domain. However, despite apparent flexibility between domains, comparison with other structures indicates conservation of a number of key interactions that introduce rigidity to the system, allowing allosteric control of the filament by interaction with ATP. Additional analysis reveals that the interaction specificity of the five human Rad51 paralogues can be predicted using a simple model based on the RadA structure.

Ways of telling : research methodology for fashion textiles design

Research methodology in the discipline of Art & Design has been a topic for much debate in the academic community. The result of such avid and ongoing discussion appears to be a disciplinary obsession with research methodologies and a culture of adopting and adapting existing methodologies from more established disciplines. This has eventuated as a means of coping with academic criticism and as an attempt to elevate Art & Design to a ‘real academic status’. Whilst this adoption has had some effect in tempering the opinion of Art & Design research from more ‘serious’ academics the practice may be concealing a deeper problem for this discipline. Namely, that knowledge transfer within creative practice, particularly in fashion textiles design practice, is largely tacit in nature and not best suited to dissemination through traditional means of academic writing and publication. ----- ----- There is an opportunity to shift the academic debate away from appropriate (or inappropriate) use of methodologies and theories to demonstrate the existence (or absence) of rigor in creative practice research. In particular, the changing paradigms for the definitions of research to support new models for research quality assessment (such as the RAE in the United Kingdom and ERA in Australia) require a re-examination of the traditions of academic writing and publication in relation to this form of research. It is now appropriate to test the limits of tacit knowledge. It has been almost half a century since Michael Polanyi wrote “we know more than we can tell” (Polanyi, 1967 p.4) at a time when the only means of ‘telling’ was through academic writing and publishing in hardcopy format. ----- ----- This paper examines the academic debate surrounding research methodologies for fashion textiles design through auto-ethnographic case study and object analysis. The author argues that, while this debate is interesting, the focus should be to ask: are there more effective ways for creative practitioner researchers to disseminate their research? The aim of this research is to examine the possibilities of developing different, more effective methods of ‘telling’ to support the transfer of tacit knowledge inherent in the discipline of Fashion Textiles Design.

Hierarchical Bayesian models for estimating the extent of plant pest invasions

Plant biosecurity requires statistical tools to interpret field surveillance data in order to manage pest incursions that threaten crop production and trade. Ultimately, management decisions need to be based on the probability that an area is infested or free of a pest. Current informal approaches to delimiting pest extent rely upon expert ecological interpretation of presence / absence data over space and time. Hierarchical Bayesian models provide a cohesive statistical framework that can formally integrate the available information on both pest ecology and data. The overarching method involves constructing an observation model for the surveillance data, conditional on the hidden extent of the pest and uncertain detection sensitivity. The extent of the pest is then modelled as a dynamic invasion process that includes uncertainty in ecological parameters. Modelling approaches to assimilate this information are explored through case studies on spiralling whitefly, Aleurodicus dispersus and red banded mango caterpillar, Deanolis sublimbalis. Markov chain Monte Carlo simulation is used to estimate the probable extent of pests, given the observation and process model conditioned by surveillance data. Statistical methods, based on time-to-event models, are developed to apply hierarchical Bayesian models to early detection programs and to demonstrate area freedom from pests. The value of early detection surveillance programs is demonstrated through an application to interpret surveillance data for exotic plant pests with uncertain spread rates. The model suggests that typical early detection programs provide a moderate reduction in the probability of an area being infested but a dramatic reduction in the expected area of incursions at a given time. Estimates of spiralling whitefly extent are examined at local, district and state-wide scales. The local model estimates the rate of natural spread and the influence of host architecture, host suitability and inspector efficiency. These parameter estimates can support the development of robust surveillance programs. Hierarchical Bayesian models for the human-mediated spread of spiralling whitefly are developed for the colonisation of discrete cells connected by a modified gravity model. By estimating dispersal parameters, the model can be used to predict the extent of the pest over time. An extended model predicts the climate restricted distribution of the pest in Queensland. These novel human-mediated movement models are well suited to demonstrating area freedom at coarse spatio-temporal scales. At finer scales, and in the presence of ecological complexity, exploratory models are developed to investigate the capacity for surveillance information to estimate the extent of red banded mango caterpillar. It is apparent that excessive uncertainty about observation and ecological parameters can impose limits on inference at the scales required for effective management of response programs. The thesis contributes novel statistical approaches to estimating the extent of pests and develops applications to assist decision-making across a range of plant biosecurity surveillance activities. Hierarchical Bayesian modelling is demonstrated as both a useful analytical tool for estimating pest extent and a natural investigative paradigm for developing and focussing biosecurity programs.

Queer/error : gay media systems and processes of abjection

Gay community media functions as a system with three nodes, in which the flows of information and capital theoretically benefit all parties: the gay community gains a sense of cohesion and citizenship through media; the gay media outlets profit from advertisers’ capital; and advertisers recoup their investments in lucrative ‘pink dollar’ revenue. But if a necessary corollary of all communication systems is error or noise, where—and what—are the errors in this system? In this paper we argue that the ‘error’ in the gay media system is Queerness, and that the gay media system ejects (in a process of Kristevan abjection) these Queer identities in order to function successfully. We examine the ways in which Queer identities are excluded from representation in such media through a discourse and content analysis of The Sydney Star Observer (Australia’s largest gay and lesbian paper). First, we analyse the way Queer bodies are excluded from the discourses that construct and reinforce both the ideal gay male body and the notions of homosexual essence required for that body to be meaningful. We then argue that abject Queerness returns in the SSO’s discourses of public health through the conspicuous absence of the AIDS-inflicted body (which we read as the epitome of the abject Queer), since this absence paradoxically conjures up a trace of that which the system tries to expel. We conclude by arguing that because the ‘Queer error’ is integral to the SSO, gay community media should practise a politics of Queer inclusion rather than exclusion.

Genome-wide identification and expression analysis of the NF-Y family of transcription factors in Triticum aestivum

Nuclear Factor Y (NF-Y) is a trimeric complex that binds to the CCAAT box, a ubiquitous eukaryotic promoter element. The three subunits NF-YA, NF-YB and NF-YC are represented by single genes in yeast and mammals. However, in model plant species (Arabidopsis and rice) multiple genes encode each subunit providing the impetus for the investigation of the NF-Y transcription factor family in wheat. A total of 37 NF-Y and Dr1 genes (10 NF-YA, 11 NF-YB, 14 NF-YC and 2 Dr1) in Triticum aestivum were identified in the global DNA databases by computational analysis in this study. Each of the wheat NF-Y subunit families could be further divided into 4-5 clades based on their conserved core region sequences. Several conserved motifs outside of the NF-Y core regions were also identified by comparison of NF-Y members from wheat, rice and Arabidopsis. Quantitative RT-PCR analysis revealed that some of the wheat NF-Y genes were expressed ubiquitously, while others were expressed in an organ-specific manner. In particular, each TaNF-Y subunit family had members that were expressed predominantly in the endosperm. The expression of nine NF-Y and two Dr1 genes in wheat leaves appeared to be responsive to drought stress. Three of these genes were up-regulated under drought conditions, indicating that these members of the NF-Y and Dr1 families are potentially involved in plant drought adaptation. The combined expression and phylogenetic analyses revealed that members within the same phylogenetic clade generally shared a similar expression profile. Organ-specific expression and differential response to drought indicate a plant-specific biological role for various members of this transcription factor family.

Linking ambulance, emergency department and hospital admissions data : understanding the emergency journey

Objective: to assess the accuracy of data linkage across the spectrum of emergency care in the absence of a unique patient identifier, and to use the linked data to examine service delivery outcomes in an emergency department setting. Design: automated data linkage and manual data linkage were compared to determine their relative accuracy. Data were extracted from three separate health information systems: ambulance, ED and hospital inpatients, then linked to provide information about the emergency journey of each patient. The linking was done manually through physical review of records and automatically using a data linking tool (Health Data Integration) developed by the CSIRO. Match rate and quality of the linking were compared. Setting: 10, 835 patient presentations to a large, regional teaching hospital ED over a two month period (August-September 2007). Results: comparison of the manual and automated linkage outcomes for each pair of linked datasets demonstrated a sensitivity of between 95% and 99%; a specificity of between 75% and 99%; and a positive predictive value of between 88% and 95%. Conclusions: Our results indicate that automated linking provides a sound basis for health service analysis, even in the absence of a unique patient identifier. The use of an automated linking tool yields accurate data suitable for planning and service delivery purposes and enables the data to be linked regularly to examine service delivery outcomes.

Developing trust networks based on user tagging information for recommendation making

Recommender systems are one of the recent inventions to deal with ever growing information overload. Collaborative filtering seems to be the most popular technique in recommender systems. With sufficient background information of item ratings, its performance is promising enough. But research shows that it performs very poor in a cold start situation where previous rating data is sparse. As an alternative, trust can be used for neighbor formation to generate automated recommendation. User assigned explicit trust rating such as how much they trust each other is used for this purpose. However, reliable explicit trust data is not always available. In this paper we propose a new method of developing trust networks based on user’s interest similarity in the absence of explicit trust data. To identify the interest similarity, we have used user’s personalized tagging information. This trust network can be used to find the neighbors to make automated recommendations. Our experiment result shows that the proposed trust based method outperforms the traditional collaborative filtering approach which uses users rating data. Its performance improves even further when we utilize trust propagation techniques to broaden the range of neighborhood.

Trust and reputation management in web-based social network

know personally. They also communicate with other members of the network who are the friends of their friends and may be friends of their friend’s network. They share their experiences and opinions within the social network about an item which may be a product or service. The user faces the problem of evaluating trust in a service or service provider before making a choice. Opinions, reputations and ecommendations will influence users' choice and usage of online resources. Recommendations may be received through a chain of friends of friends, so the problem for the user is to be able to evaluate various types of trust recommendations and reputations. This opinion or ecommendation has a great influence to choose to use or enjoy the item by the other user of the community. Users share information on the level of trust they explicitly assign to other users. This trust can be used to determine while taking decision based on any recommendation. In case of the absence of direct connection of the recommender user, propagated trust could be useful.