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To investigate the patterns of ED use in metropolitan and rural New South Wales (NSW) by socioeconomic status (SES).We undertook a retrospective, population-based study of de-identified data from the NSW Emergency Department Data Collection (EDDC). The study population comprised of NSW residents who presented to an NSW public hospital ED in 2013-2019 and were registered in the NSW EDDC. Total ED presentations, negative binomial regression modelled annual changes in ED presentations over 2013-2019, and age- and sex-standardised rates of ED presentations in 2019 were assessed.Overall, between 2013 and 2019, ED presentations increased in metropolitan and rural NSW, with mean annual percentage increases of 3.1% (95% confidence interval [CI] 2.8-3.5) and 2.5% (95% CI 2.0-2.9), respectively. This growth varied by SES, with larger increases observed in higher SES groups. The bulk of presentations in rural NSW were from individuals living in disadvantaged areas. Standardised rates of ED presentations were highest in the most disadvantaged quintiles (SES 1) and progressively decreased with increasing SES in both rural and metropolitan NSW (negative gradients). Rates were higher in rural NSW compared to metropolitan NSW across all SES quintiles for total, low acuity and non-low acuity presentations.Negative gradients in rates of ED presentations with increasing SES were observed in both metropolitan and rural NSW. At each SES quintile, rates of ED presentations were higher in rural compared to metropolitan areas. Further research exploring the underlying causal mechanisms leading to increased ED demand in rural NSW and socioeconomically disadvantaged populations is warranted.

Australia is facing a growing burden of knee and hip osteoarthritis (OA). To address this demand in northern New South Wales, a community health-based conservative OA joint management service was established in the Tweed Valley. This paper describes the design, implementation and initial evaluation of the service. Following the principles of clinical redesign, a diagnostic phase involving consultation with key stakeholders revealed several issues. OA patients could wait up to 9 months for review by orthopaedic specialist following GP referral and received limited information on how to conservatively manage their conditions. GPs were constrained by short consultations and had limited knowledge of the latest recommendations for the conservative treatment of OA. GPs also highlighted the limitations of outdated fax systems for communication, noting their preference for secure electronic messaging. Based on these findings, the Tweed Knee and Hip Arthritis Service was established. For patients not on a waiting list for surgery, the service provides evidence-based conservative management for knee or hip OA involving standardised assessment, education, exercise, self-management strategies and regular review. An analysis of a foundational cohort of patients demonstrated improvements in a suite of validated and standardised measures for pain and function, with improvements seen as early as 1 month and sustained for 6 months. The study findings support the introduction of integrated conservative OA management models of care directly available to primary healthcare providers.

Objective To explore the patterns of and investigate the factors associated with rises in emergency department presentations in rural and metropolitan New South Wales from 2012 to 2018. Design A retrospective descriptive study of de-identified data from the New South Wales Emergency Department Data Collection. Setting New South Wales, Australia. Participants All individuals presenting to 99 New South Wales emergency departments, which continuously reported to the Emergency Department Data Collection between 2012 and 2018. A total of 2 166 449 presentations recorded throughout New South Wales in 2012 (rural 786 278; metropolitan 1 380 171) and 2 477 192 in 2018 (rural 861 761; metropolitan 1 615 431). Main outcome measures Total emergency department presentations, plus Poisson regression modelled annual changes in emergency department presentations over the period 2012-2018. Results Growth in emergency department presentations outpaced population growth in both rural and metropolitan New South Wales between 2012 and 2018. The patterns of age-standardised rates of presentations were broadly similar between rural and metropolitan areas, with highest rates observed in the youngest (0-4 years) and oldest (85+ years) cohorts. The rural sample also displayed a distinct third peak in ages 15-39 years, and rates were higher across all age groups. Rural New South Wales displayed disproportionately higher emergency department presentations in the two most deprived socio-economic status quintiles. While rural New South Wales displayed significant reductions in triage category 5 (non-urgent cases) over time, the relative proportion remained approximately double that of metropolitan sites. Conclusions There are differences between rural and metropolitan emergency department presentations relating to demographic factors, triage levels, acuity and admissions. Detailed local investigations are required to determine specific contextual issues that impact on emergency department demand.

BackgroundWe conducted a survey to identify what types of health research could be exempted from research ethics reviews in Australia.MethodsWe surveyed active Australian health researchers and members of Human Research Ethics Committees (HREC). We presented the respondents with eight hypothetical research scenarios, involving: N of 1 trials, no treatment studies, linked data sets, surplus samples, audits, surveys, interviews with patients, and professional opinion. We asked whether these scenarios should or should not be exempt from ethics review, and to provide (optional) explanations. We analysed the reasons thematically, to identify Top 3 reasons underlying the decisions.ResultsMost frequent reasons for requiring ethics reviews, included: the need for independent oversight, privacy/confidentiality issues, review of scientific rigour, and publishing considerations. Most frequent reasons for exempting scenarios from reviews, included: level of risk, study design, privacy/confidentiality issues, and standard clinical practice. Four research scenarios listed the same Top 3 reasons for requiring ethics reviews: need for independent oversight, review of scientific rigour, privacy/confidentiality. Reasons for exempting were less uniform, but low risk was a Top 3 reason for 7 scenarios, and study design for 4 scenarios. Privacy/confidentiality was given as a Top 3 reason for both requiring and exempting from ethics the same two scenarios.ConclusionsThe most frequently offered reasons in support of requiring ethics reviews for research scenarios are more uniform than those for exempting them. However, considerable disagreement exists about when the risks of research are so minimal that the exemption is appropriate.

BackgroundWe conducted a survey to identify what types of health/medical research could be exempt from research ethics reviews in Australia.MethodsWe surveyed Australian health/medical researchers and Human Research Ethics Committee (HREC) members. The survey asked whether respondents had previously changed or abandoned a project anticipating difficulties obtaining ethics approval, and presented eight research scenarios, asking whether these scenarios should or should not be exempt from ethics review, and to provide (optional) comments. Qualitative data were analysed thematically; quantitative data in R.ResultsWe received 514 responses. Forty-three per cent of respondents to whom the question applied, reported changing projects in anticipation of obstacles from the ethics review process; 25% reported abandoning projects for this reason. Research scenarios asking professional staff to provide views in their area of expertise were most commonly exempted from ethics review (to prioritise systematic review topics 84%, on software strengths/weaknesses 85%); scenarios involving surplus samples (82%) and N-of-1 (single case) studies (76%) were most commonly required to undergo ethics review. HREC members were 26% more likely than researchers to require ethics review. Need for independent oversight, and low risk, were most frequently cited in support of decisions to require or exempt from ethics review, respectively.ConclusionsConsiderable differences exist between researchers and HREC members, about when to exempt from review the research that ultimately serves the interests of patients and the public. It is widely accepted that evaluative research should be used to reduce clinical uncertainties—the same principle should apply to ethics reviews.

Australia is facing a growing burden of knee and hip osteoarthritis (OA). To address this demand in northern New South Wales, a community health-based conservative OA joint management service was established in the Tweed Valley. This paper describes the design, implementation and initial evaluation of the service. Following the principles of clinical redesign, a diagnostic phase involving consultation with key stakeholders revealed several issues. OA patients could wait up to 9 months for review by orthopaedic specialist following GP referral and received limited information on how to conservatively manage their conditions. GPs were constrained by short consultations and had limited knowledge of the latest recommendations for the conservative treatment of OA. GPs also highlighted the limitations of outdated fax systems for communication, noting their preference for secure electronic messaging. Based on these findings, the Tweed Knee and Hip Arthritis Service was established. For patients not on a waiting list for surgery, the service provides evidence-based conservative management for knee or hip OA involving standardised assessment, education, exercise, self-management strategies and regular review. An analysis of a foundational cohort of patients demonstrated improvements in a suite of validated and standardised measures for pain and function, with improvements seen as early as 1 month and sustained for 6 months. The study findings support the introduction of integrated conservative OA management models of care directly available to primary healthcare providers.

ObjectivesTo identify what types of health and medical research could be exempted from a requirement to undergo research ethics reviews in Australia.DesignA web-based survey.SettingAustralian-based participants completed the survey online between 10 September and 1 November 2019.ParticipantsActive human health and medical researchers and members of Human Research Ethics Committees (HREC).Main outcome measuresParticipants were asked whether they self-identify as a researcher, a member of an Australian human research ethics committee, or both. They were asked whether they had previously changed or abandoned a research project in anticipation of difficulty in obtaining ethics approval; and were presented with 4 hypothetical research scenarios, asking whether they should or should not be exempt from ethics review in Australia. All but the demographic question allowed the respondent to provide a comment clarifying their response. Qualitative data were analysed thematically; quantitative data were analysed in R.ResultsWe received 514 survey responses: 153 from researchers, 196 from HREC members, and 163 from individuals who identified as both. 27% of researchers reported they had changed their projects because they anticipated obstacles resulting from the ethics review process, and 16% abandoned projects for this reason. The most commonly exempted research scenarios involved professional staff providing their views on their area of expertise (84%, 85%); scenarios involving surplus samples and N-of-1 studies in clinical practice were most commonly required to undergo ethics review (82%, 76% respectively). Probability of answering that ethics review was required was the highest for HREC members and nearly identical for respondents who were both HREC members and researchers; it was the lowest for researchers. HREC members were overall 26% more likely to answer that ethics review was required than researchers.ConclusionsOur survey shows considerable differences between the researchers and members of ethics committees, about how best to serve the interests of patients and the public. Those interests are best served by evaluative research to reduce or resolve the uncertainties in the clinical context – the same general principle should be applied to ethics review.

Objectives: To determine age group‐ and cause‐of‐death‐specific contributions to area socioeconomic status (SES), sex and remoteness life expectancy inequalities. Methods: Mortality and estimated residential population data from New South Wales, Australia, over 2010–2012 was used to calculate life expectancy. Inequalities by sociodemographic groups were partitioned into age group‐ and cause‐of‐death‐specific contributions. Results: The largest contributions to SES differentials in life expectancy were observed at 60–84 years of age; for cancer, cardiovascular, endocrine and respiratory causes of death; and additionally external causes of death for males. Sex inequalities ranged from 3.6 to 5.2 years, with common causes of death such as cardiovascular disease and cancer in late adulthood (60+ years) accounting for the bulk of the differences. Smaller differences in life expectancy were observed by remoteness, with the largest contributions observed in ages 85 years and above, and for cardiovascular, mental, cancer and external causes of death. Conclusions: Common causes of death in late adulthood accounted for the bulk of life expectancy inequalities. Implications for public health: Development of policy and interventions aimed at addressing social determinants, such as proposed by the WHO's Global Plan of Action, are needed to help reduce sociodemographic inequalities in lifespan.

Category: Diabetes Introduction/Purpose: Diabetic ulcers carry with it extensive morbidity. The microbiome of diabetic foot infections have previously been shown to be polymicrobial. We hypothesized that ulcer treatment would normalize the foot microbiome. Methods: Microbiome profiles of the fore, mid and hind diabetic foot. Diabetic foot (with ulcer) were taken pre and post treatment with a combination of surgical debridement, local and systemic antibiotics. 10 subjects were studied. Microbiome graphs and images pre and post treatment of feet are shown. Results: The microbiome profile of the diabetic foot almost completely normalized to that of normal flora. Before treatment, the foot microbiome was associated with polymicrobial pathogenic bacteria mostly away from the ulcer. The ulcer itself only contained minimal species, albeit pathogenic bacteria. The differences in profiles between the pre and post treatment were significantly different p < 0.01. Conclusion: Chicken or egg, the foot microbiome almost completely reverts to normal flora with simple ulcer debridement and local/systemic antibiotics. Our data point possibly suggest ulcers could be prevented by reducing pathogenic microbiome load (chlorhexidine washes) or microbiome transplants. We next move to controls as a proof of concept.

This article describes the evaluation of enhanced surveillance that was conducted on an older age-group of children with pertussis (ages 5-9 years), in addition to the routine follow-up of a younger age-group (ages 0-4 years) in Sydney Local Health District during 2015. We found little incremental benefit in this policy in terms of identifying additional high-risk contacts, despite a greatly increased workload.

ObjectivesLevel of education and genetic risk are key predictors of cardiovascular disease (CVD). While several studies have explored the causal mechanisms of education effects, it remains uncertain to what extent genetic risk is mediated by established CVD risk factors. This study sought to investigate this and explored the mediation of education and genetic effects on CVD by established cardiovascular risk factors in the Framingham Heart Study (FHS).DesignProspective observational cohort study.Participants7017 participants from the FHS.SettingCommunity-based cohort of adults in Framingham, Massachusetts, USA.Primary outcome measureIncident CVD. The total effects of education and genetic predisposition using a 63-variant genetic risk score (GRS) on CVD, as well as those mediated by established CVD risk factors, were assessed via mediation analysis based on the counterfactual framework using Cox proportional hazards regression models.ResultsOver a median follow-up time of 12.0 years, 1091 participants experienced a CVD event. Education and GRS displayed significant associations with CVD after adjustment for age and sex and the established risk factors smoking, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), body mass index, systolic blood pressure (SBP) and diabetes. For education effects, smoking, HDL-C and SBP were estimated to mediate 18.8% (95% CI 9.5% to 43%), 11.5% (95% CI 5.7% to 29.0%) and 4.5% (95% CI 1.6% to 13.3%) of the total effect of graduate degree, respectively, with the collective of all risk factors combined mediating 38.5% (95% 24.1% to 64.9%). A much smaller proportion of the effects of GRS were mediated by established risk factors combined (17.6%, 95% CI 2.4% to 35.7%), with HDL-C and TC mediating 11.5% (95% CI 6.2% to 21.5%) and 3.1% (95% CI 0.2% to 8.3%), respectively.ConclusionsUnlike education inequalities, established risk factors mediated only a fraction of GRS effects on CVD. Further research is required to elucidate the underlying causal mechanisms of genetic contributions to CVD.

Objectives To assess the association between ED occupancy and relevant outcomes including ED waiting times, rates of admission and representation and length of stay when hospitalised. Methods Retrospective study of all ED presentations by New South Wales (NSW), Australia, residents to 15 NSW public, principal referral or paediatric specialist hospitals between 1 January to 31 December 2015 (N = 935 282). ED data were linked longitudinally (to ED data) and cross-sectionally to hospital admissions data. An ED-system measure of occupancy was assigned to each ED record. The study outcomes were ED waiting time, admission to hospital, 28 day representation, and length of stay (LOS) when admitted. Outcomes were analysed using univariate analyses and multivariable general linear and binary logistic regression models. Results Increased ED occupancy was associated with increased ED waiting times, particularly at low-baseline occupancy (e.g. rate ratio = 2.22, 95% confidence interval [CI] [2.10-2.35], for non-urgent triaged patients). However, results were conditional on triage category, such that estimated effects were smaller or not significant in emergency and resuscitation triaged patients (e.g. rate ratio = 1.59, 95% CI [1.52-1.65], for emergency patients). ED occupancy only showed small or no associations with admission to hospital, 28 day representation and LOS when admitted. Conclusions Higher ED occupancy was associated with increased waiting times conditional on triage category and baseline occupancy. Collectively, the results show that NSW principal referral EDs are robust, and are currently capable of handling variation in occupancy by prioritising treatment for the most urgent patients.

Introduction Currently there are no clinical screening tests available to identify pregnancies at risk of developing preeclampsia (PET) and/or intrauterine growth restriction (IUGR), both of which are associated with abnormal placentation. Metabolic profiling is now a stable analytical platform used in many laboratories and has successfully been used to identify biomarkers associated with various pathological states. Methods We used nuclear magnetic resonance spectroscopy (NMR) to metabolically profile serum samples collected from 143 pregnant women at 26–41 weeks gestation with pregnancy outcomes of PET, IUGR, PET IUGR or small for gestational age (SGA) that were age-matched to normal pre/term pregnancies. Results Spectral analysis found no difference in the measured metabolites from normal term, pre-term and SGA samples, and of 25 identified metabolites, only glutamate was marginally different between groups. Of the identified metabolites, 3-methylhistidine, creatinine, acetyl groups and acetate, were determined to be independent predictors of PET and produced area under the curves (AUC) = 0.938 and 0.936 for the discovery and validation sets. Only 3-hydroxybutyrate was determined to be an independent predictor of IUGR, however the model had low predictive power (AUC = 0.623 and 0.581 for the discovery and validation sets). Conclusions A sub-panel of metabolites had strong predictive power for identifying PET samples in a validation dataset, however prediction of IUGR was more difficult using the identified metabolites. NMR based metabolomics can identify metabolites strongly associated with disease and has the potential to be useful in developing early clinical screening tests for at risk pregnancies.

Background To investigate whether the adverse infant health outcomes associated with early birth and severe neonatal morbidity (SNM) persist beyond the first year of life and impact on paediatric hospitalisations for children up to 6 years of age. Methods The study population included all singleton live births, >32 weeks gestation in New South Wales, Australia, in 2001–2005, with follow-up to 6 years of age. Birth data were probabilistically linked to hospitalisation data (n = 392 964). The odds of hospitalisation, mean hospital length of stay (LOS) and costs, and cumulative LOS were evaluated by gestational age and SNM using multivariable analyses. Results A total of 74 341 (18.9%) and 41 404 (10.5%) infants were hospitalised once and more than once, respectively. SNM was associated with increased odds of hospitalisation once (adjusted odds ratio [aOR] 1.16 [95% confidence interval 1.10, 1.22]) and more than once [aOR 1.51 (1.43, 1.61)]. Decreasing gestational age was associated with increasing odds of hospitalisation more than once from aOR 1.19 at 37–38 weeks to 1.49 at 33–34 weeks. Average LOS and costs per hospital admission were increased with SNM but not with decreasing gestational age. Cumulative LOS was significantly increased with SNM and decreasing gestational age. Conclusions Adverse effects of SNM and early birth persist between 1 and 6 years of age. Strategies to prevent early birth and reduce SNM, and to increase health monitoring of vulnerable infants throughout childhood may help reduce paediatric hospitalisations.

Summary We determined brain to liver weight ratio (BLWR) thresholds for fetal growth restriction (FGR) using autopsy information on 395 perinatal deaths comprising stillborn babies who died during labour and neonatal deaths. FGR was defined using two methods: (1) birth weight for gestational age (WGA) less than the 10th percentile; and (2) WGA less than the 10th percentile or discordant birth weight/length. The association between BLWR and FGR was investigated using odds ratios, and classification statistics were calculated for a range of BLWR thresholds. Using WGA, 84 cases (21.3%) were FGR and a further 15 cases (n = 99, 25%) had discordant birth weight/length. The BLWR ranged from 1.02 to 7.30 and was positively associated with FGR. BLWR was not associated with FGR for babies with congenital central nervous system or chromosomal abnormalities. Excluding these, for FGR defined using WGA and discordant birth weight/length, a BLWR threshold of 5.0 was 100% predictive of FGR. A BLWR threshold of 3.0 for babies over 28 weeks gestation and 3.7 for more preterm babies optimised case detection while minimising missed and false positive cases. Additional evidence of FGR should be sought for babies with a BLWR of less than 5.0 to confirm FGR.

Objectives Despite being one of the healthiest countries in the world, Australia displays substantial mortality differentials by socioeconomic disadvantage, remoteness and sex. In this study, we examined how these mortality differentials translated to differences in life expectancy between 2001 and 2012. Design and setting Population-based study using mortality and estimated residential population data from Australia9s largest state, New South Wales (NSW), between 2001 and 2012. Age-group-specific death rates by socioeconomic disadvantage quintile, remoteness (major cities vs regional and remote areas), sex and year were estimated via Poisson regression, and inputted into life table calculations to estimate life expectancy. Results Life expectancy decreased with increasing socioeconomic disadvantage in males and females. The disparity between the most and least socioeconomically deprived quintiles was 3.77 years in males and 2.39 years in females in 2012. Differences in life expectancy by socioeconomic disadvantage were mostly stable over time. Gender gaps in life expectancy ranged from 3.50 to 4.93 years (in 2012), increased with increasing socioeconomic disadvantage and decreased by ∼1 year for all quintiles between 2001 and 2012. Overall, life expectancy varied little by remoteness, but was 1.8 years higher in major cities compared to regional/remote areas in the most socioeconomically deprived regions in 2012. Conclusions Socioeconomic disadvantage and sex were strongly associated with life expectancy. The disparity in life expectancy across the socioeconomic spectrum was larger in males and was stable over time. In contrast, gender gaps reduced for all quintiles between 2001 and 2012, and a remoteness effect was evident in 2012, but only for those living in the most deprived areas.

The RUNX2 transcription factor is indispensable for skeletal development and controls bone formation by acting as a signaling hub and transcriptional regulator to coordinate target gene expression. A signaling partner of RUNX2 is the nuclear vitamin D receptor (VDR) that becomes active when bound by its ligand 1,25-dihydroxyvitamin D3 (VD3). RUNX2 and VDR unite to cooperatively regulate the expression of numerous genes. In this study, we overexpressed RUNX2 in NIH3T3 fibroblasts concomitantly treated with VD3 and show that RUNX2 alone, or in combination with VD3, failed to promote an osteoblastic phenotype in NIH3T3 cells. However, the expression of numerous osteoblast-related genes was up-regulated by RUNX2 and large-scale gene expression profiling using microarrays identified over 800 transcripts that displayed a twofold of greater change in expression in response to RUNX2 overexpression or VD3 treatment. Functional analysis using gene ontology (GO) revealed GO terms for ossification, cellular motility, biological adhesion, and chromosome organization were enriched in the pool of genes regulated by RUNX2. For the set of genes whose expression was modulated by VD3, the GO terms response to hormone stimulus, chemotaxis, and metalloendopeptidase activity where overrepresented. Our study provides a functional insight into the consequences of RUNX2 overexpression and VD3 treatment in NIH3T3 cells in addition to identifying candidate genes whose expression is controlled by either factor individually or through their functional cooperation.

Research capacity building in healthcare works to generate and apply new knowledge to improve health outcomes; it creates new career pathways, improves staff satisfaction, retention and organisational performance. While there are examples of investment and research activity in rural Australia, overall, rural research remains under-reported, undervalued and under-represented in the evidence base. This is particularly so in primary care settings. This lack of contextual knowledge generation and translation perpetuates rural–metropolitan health outcome disparities. Through greater attention to and investment in building research capacity and capability in our regional, rural and remote health services, these issues may be partially addressed. It is proposed that it is time for Australia to systematically invest in rurally focussed, sustainable, embedded research capacity building.

A new N-acetyl-D-glucosamine (GlcNAc) specific lectin was identified and purified from the fruiting body of the Australian indigenous mushroom Psathyrella asperospora. The functional lectin, named PAL, showed hemagglutination activity against neuraminidase treated rabbit and human blood types A, B and O, and exhibited high binding specificity towards GlcNAc, as well as mucin and fetuin, but not against asialofetuin. PAL purified to homogeneity by a combination of ammonium sulfate precipitation, chitin affinity chromatography and size exclusion chromatography, was monomeric with a molecular mass of 41.8 kDa, was stable at temperatures up to 55 °C and between pH 6–10, and did not require divalent cations for optimal activity. De novo sequencing of PAL using LC-MS/MS, identified 10 tryptic peptides that revealed substantial sequence similarity to the GlcNAc recognizing lectins from Psathyrella velutina (PVL) and Agrocybe aegerita (AAL-II) in both the carbohydrate binding and calcium binding sites. Significantly, PAL was also found to exert a potent anti-proliferative effect on HT29 cells (IC50 0.48 μM) that was approximately 3-fold greater than that observed on VERO cells; a difference found to be due to the differential expression of cell surface GlcNAc on HT29 and VERO cells. Further characterization of this activity using propidium iodine staining revealed that PAL induced cell cycle arrest at G2/M phase in a manner dependent on its ability to bind GlcNAc.

Background Although infant and child mortality rates have decreased substantially worldwide over the past two decades, efforts continue in many nations to further these declines. The identification of pertinent perinatal factors that are associated with early childhood mortality would help with these efforts. We investigated the association of two crucial perinatal factors, gestational age and severe neonatal morbidity at birth, with mortality during infancy (29-364 days) and early childhood (1-5 years). Methods The study population included all singleton livebirths, ≥32 weeks' gestation in New South Wales, Australia in 2001-11. Birth data were linked to hospitalisation morbidity data and deaths data (linked birth cohort n = 871 916), and multivariable Cox regression models were used to assess mortality. Results The median follow-up time per child was 4.95 years (range 0.00-5.92 years; 3 614 738 total person-years), with 984 deaths observed. Gestational age was associated with increased mortality, and specifically from deaths attributable to infections, respiratory conditions, and injuries during infancy, but not during early childhood. Severe neonatal morbidity strongly mediated the effects of gestational age during infancy, but not during early childhood, and was associated with increased mortality from circulatory, nervous, and respiratory system causes. Conclusions The direct effects of gestational age on mortality extended up to 1 year of age, whereas severe neonatal morbidity remained associated with heightened mortality into early childhood. Efforts to maximise the health and well-being of vulnerable infants, with emphasis on preventing infections and injuries, may help further reduce early childhood mortality.

Objective To explore the patterns of low acuity patient (LAP) presentations to EDs in New South Wales (NSW), Australia. Methods Retrospective study of NSW public hospital ED presentations between January 2013 and December 2014 that were registered in the NSW Emergency Department Data Collection (n = 409 035). LAPs were defined according to the Australian Institute of Health and Welfare (AIHW), Sprivulis and multiple ACEM methods. Multivariable logistic regression was used to assess the adjusted odds of LAP ED presentation by a suite of sociodemographic factors. Results The percentage of LAPs varied considerably by definition, being as high as 54.7% (inner regional areas) and as low as 3.2% (major cities) using revised ACEM methods modified to contain unlimited consultation times or consultation times of 15 min or less, respectively. For each method, higher proportions of LAPs were observed in inner regional and remote/very remote areas relative to major cities. LAP ED presentations, based on ACEM definition with 1 h or 15 min consultation times, were greater in younger patients, increased during out of business hours and weekends, and decreased with increasing general practitioner (GP) density. Conclusion The percentage of LAPs varied substantially by definition, and further work is required to validate the methods, particularly around the appropriateness of length of consultation time with ACEM, between different hospitals and remoteness areas. Age was strongly associated with low acuity, with substantial effects also observed for GP density, and attendances during out of hours and weekends.

Cell-based assays represent a major end point of high throughput screening (HTS) but a key limitation of such assays is the potentially poor membrane permeability of test compounds. In this study, we optimized the conditions for the delivery of the membrane impermeable compound 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) into human cells using hypotonic shift; a method that can promote the uptake of molecules from the extracellular fluid into cell cytoplasm via endocytosis. We showed that uptake of HPTS by cells was a function of hypotonic buffer osmolarity and that delivery was highly efficient with almost 100% of cells displaying uptake. Delivery of HPTS was equally effective at 25°C and 37°C, with delivery of compound proportional to incubation time and concentration of HPTS within the loading medium. The experimental conditions identified in this study could be applied to HTS drug discovery studies providing an effective method of delivering small membrane impermeable compounds into cells.

The RUNX2 transcription factor is essential for osteoblast differentiation and chondrocyte maturation. Rare mutations within RUNXZ cause the dominantly inherited skeletal disorder cleidocranial dysplasia which is charaterized by gross dysgenesis of the skeleton. A unique feature of RUNX2 is the polyglutamine and polyalanine (poly Q/A)domain encoded by a trinucleotide repeat sequence. Such repeat sequences are prone to strand slippage resulting in high mutation rates. We hypothesized that mutations within the RUNX2 poly Q/A domain would exist and these variants would be associated with altered bone density and protein function. 4361 DNA samples were obtained from multiple epidemiological studies of bone density. Bone density was measured by DEXA and ultrasound and expressed as Z-scores around the appropriate age-adjusted mean. A total of 21 subjects were identified as being heterozygous for a wild type 23Q117A allele and a p l y Q/A repeat variant allele. Deletions (ISQ and 164) and insertions (304 and 23A) were identified. Collectively Q/A repeat variants presented with significantly lower Femoral Neck BMD (DEXA) displaying a 0.65 SD decrease (n = 21, p = 0.0004) and lower bone density as measured by quantitative ultrasound of similar magnitude (-0.79 SD, n = 8, p = 0.031). To understand the mechanism via which the rare mutations were decreasing bone density, functional analyses were carried out on 23Q (WT), 16Q and 304 RUNX2. The ability of the proteins to bind to the RUNX2 DNA binding site was assessed by EMSA. The analysis did not reveal any obvious changes in the DNA binding capacities of the mutant proteins. However, reporter gene analysis using the mouse osteocalcin promoter revealed significant decreases in the transactivation function of 169 and 30Q. Finally, the ability of the proteins to localise to the nucleus was assessed. Subcellular analysis revealed the 164 and 30Q proteins displayed defective nuclear localisation compared to WT. We have identified a new class of functionally relevant RUNX2 variants that occur at collective frequency of -0.5%. These mutations are associated with significantly lower bone density and altered protein function.

Objectives To investigate survival, hospitalization, and acute-care costs of very (28-31 weeks' gestation) and moderate preterm (32-33 weeks' gestation) infants in the first 6 years of life and compare outcomes with the more widely studied extremely preterm infants (24-27 weeks' gestation) and to full term (low risk) infants (39-40 weeks' gestation). Study design Birth data from all women residing in New South Wales, Australia, with gestational ages between 24-33 and 39-40 weeks in 2001-2011 were linked probabilistically to hospitalization and mortality data. Study outcomes were evaluated with the use of descriptive and multivariable analyses at birth (N = 559 532), discharge (N = 540 240), and at 1 (N = 487 447) and 6 years of age (N = 230 498). Results Mortality was greatest among extremely preterm infants (eg, 31.2% within 6 years) and decreased with increasing gestational age. Likewise, hospitalization within the first year of life increased with decreasing gestational age (aOR 5.5 [95% CI 4.7-6.4], 3.7 [3.4-4.0], and 2.6 [2.5-2.8] for birth at 24-27, 28-31, and 32-33 weeks' gestation, relative to 39-40 weeks' gestation). Hospitalization remained significantly increased with preterm birth at each year of age up to 6 years (aORs 1.3-1.6 at 6 years). Cumulative costs were significantly greater with preterm birth within the first year of life, and also between 1 and 6 years of age. Conclusions The risks of adverse health outcomes were significantly greater in very and moderately preterm infants relative to full term infants but lower than extremely preterm infants. Crucially, preterm birth was associated with prolonged increased odds of hospitalization (up to age 6 years), contributing to greater resource use.

At present, there are no vaccines approved for the prevention or treatment of malignant melanoma, despite the amount of time and resources that has been invested. In this study, we aimed to develop a self-contained vaccine capable of directly stimulating anticancer CD8(+) T-cell immune responses. To achieve this, three whole-cell melanoma vaccines were developed expressing 4-1BBL or B7.1 T-cell co-stimulatory molecules individually or in combination. The ability of engineered vaccine cell lines to stimulate potent anticancer immune responses in C57BL/6 mice was assessed. Mice vaccinated with cells overexpressing both 4-1BBL and B7.1 (B16-F10-4-1BBL-B7.1-IFNγ/β anticancer vaccine) displayed the greatest increases in CD8(+) T-cell populations (1.9-fold increase versus control within spleens), which were efficiently activated following antigenic stimulation, resulting in a 10.7-fold increase in cancer cell cytotoxicity relative to control. The enhanced immune responses in B16-F10-4-1BBL-B7.1-IFNγ/β-vaccinated mice translated into highly efficient rejection of live tumour burdens and conferred long-term protection against repeated tumour challenges, which were likely due to enhanced effector memory T-cell populations. Similar results were observed when dendritic cell (DC)-deficient LTα(-/-) mice were treated with the B16-F10-4-1BBL-B7.1-IFNγ/β anticancer vaccine, suggesting that the vaccine can directly stimulate CD8(+) T-cell responses in the context of severely reduced DCs. This study shows that the B16-F10-4-1BBL-B7.1-IFNγ/β anticancer vaccine acted as a highly effective antigen-presenting cell and is likely to be able to directly stimulate CD8(+) T-cells, without requiring co-stimulatory signals from either CD4(+) T-cells or DCs, and warrants translation of this technology into the clinical setting.

RUNX2 gene SNPs were genotyped in subjects from the upper and lower deciles of age- and weight-adjusted femoral neck BMD. Of 16 SNPs in RUNX2 and its two promoters (P1 and P2), only SNPs in the P2 promoter were significantly associated with BMD. These P2 promoter SNPs were functionally different in gel-shift and promoter activity assays. Introduction: Specific osteoblast genes are induced by Runx2, a cell-specific transcription factor that is a candidate gene for controlling BMD. We tested the hypothesis that RUNX2 genetic variation is associated with BMD. Materials and Methods: From a population repository of normal subjects, the age- and weight-adjusted femoral neck BMD was ranked, and the upper and lower deciles (n = 132 each) were taken to represent the adjusted extremes of the population distribution. In these 264 subjects, we identified 16 allelic variations within the RUNX2 gene and promoters (P1 and P2) through DNA sequencing and denaturing high-performance liquid chromatography. Characterization of these alleles was performed through allele-specific cloning, transfection into ROS 17/2.8 cells, luciferase reporter analysis, and electrophoretic mobility shift assays. Results: Within the P2 promoter were three polymorphic nucleotides for which the minor alleles were over-represented in the upper decile of BMD (0.117 and 0.064 in the upper and lower deciles, respectively). These alleles are in near complete linkage disequilibrium with each other and represent a haplotype block that is significantly associated with increased BMD. The common and rare P2 promoter alleles were cloned upstream of luciferase, and when transfected into osteoblast-like cells, the construct representing the rare haplotype showed significantly greater P2 promoter activity than the common haplotype. Conclusions: Because the high BMD allele had higher P2 promoter activity, the data suggest that greater RUNX2 P2 promoter activity is associated with higher BMD.

We hypothesized that by taking subjects from the extremes of a population, we would specifically find alleles related to the bone mineral density trait, if such alleles exist for any particular target gene. We tested this idea using RUNX2, the well known osteoblast transcription factor. From a population of 1300 subjects the age-weight adjusted femoral neck bone mineral density (BMD) was ranked and the upper and lower deciles taken to represent the adjusted extremes. After adjusting and ranking, the two groups (n=130 each) were not significantly different for age or weight. In these 260 subjects, we identified 19 allelic variations within the RUNX2 gene and promoter (P1 and P2), and characterized these novel variations with respect to BMD strata by genotype using dHPLC. A general linkage disequilibrium (LD) analysis of RUNX2 showed that LD drops off with physical distance, as expected. Within the P2 promoter were three polymorphic nucleotides for which the minor alleles were over represented in the upper decile of BMD. These alleles are in complete LD with each other and represent a haplotype block that is significantly associated with increased BMD. Constructs made from the "normal" and the "high BMD" alleles were cloned upstream of otherwise identical luciferase constructs. Despite there being only 3 nucleotide differences between the two constructs, when transfected into osteoblast-like cells the high BMD allele construct showed significantly greater P2 promoter activity than the normal allele. Comparative homology studies showed that the high BMD alleles were the major forms in other animals (dog, rat, mouse, chimp), suggesting that the human normal BMD alleles represent the evolutionary change. The rat sequence was identical to the human high BMD allele, permitting an analysis of DNAprotein interactions in the well characterized ROS17/2.8 rat osteoblast cell line. These data showed abundant nuclear factor binding to one the three polymorphic sites in the human normal BMD allele that does not occur in the high BMD allele (and rat sequence). The data suggest that the high BMD allele does not interact with a factor that otherwise binds strongly to the normal human allele. Since the high BMD allele had higher P2 promoter activity, the binding activity may be a repressor that limits the expression of the normal allele. In any event, the nuclear factor is a candidate for the allele-specific regulation of the RUNX2 promoter.

Population ageing presents significant challenges for many developed nations. Accurately forecasting the likely future burden of age-related medical conditions, such as hip fracture, is critical. In this study, we present estimates of the current and future burden of hip fracture in NSW, Australia, providing crucial information for future health care planning. The aims of this study were to investigate the burden of hip fracture in Australia’s largest state, New South Wales (NSW), and to build a prediction model to forecast the likely future burden of hip fracture from 2016 to 2036 in persons aged 50 years or more. A retrospective population-based cohort study was conducted using NSW hospitalisation data. Standardised incident hip fracture rates and hip fracture-related acute care length of stay and costs were estimated. Predictive negative binomial regression modelling using age, gender and local health district and year covariates together with projected NSW populations was applied to forecast future hip fractures. Total incident hip fractures increased 8.8 % over a 12-year period from 2000/2001 to 2011/2012 despite declining age-standardised rates. Estimates of acute care length of stay for the treatment of hip fracture ranged from 10 to 15 days and acute care costs ranged between 21 and 29,000 Australian dollars per fracture. By 2036, incident hip fractures are projected to rise by 35.2 %, assuming a continued decline in the rate of hip fracture or by 107.5 % if the current decline in the rate does not continue. Acute care length of stay and costs are each predicted to rise between 37.1 and 110.4 % by 2036. An ageing population and changing demographics will continue to drive the increasing burden of incident hip fractures in NSW and Australia in the foreseeable future. These anticipated changes provide important information for the planning and management of future hip fracture care.

A new N-acetyl-D-glucosamine (GlcNAc) specific lectin was identified and purified from the fruiting body of the Australian indigenous mushroom Psathyrella asperospora. The functional lectin, named PAL, showed hemagglutination activity against neuraminidase treated rabbit and human blood types A, B and O, and exhibited high binding specificity towards GlcNAc, as well as mucin and fetuin, but not against asialofetuin. PAL purified to homogeneity by a combination of ammonium sulfate precipitation, chitin affinity chromatography and size exclusion chromatography, was monomeric with a molecular mass of 41.8 kDa, was stable at temperatures up to 55 °C and between pH 6-10, and did not require divalent cations for optimal activity. De novo sequencing of PAL using LC-MS/MS, identified 10 tryptic peptides that revealed substantial sequence similarity to the GlcNAc recognizing lectins from Psathyrella velutina (PVL) and Agrocybe aegerita (AAL-II) in both the carbohydrate binding and calcium binding sites. Significantly, PAL was also found to exert a potent anti-proliferative effect on HT29 cells (IC50 0.48 μM) that was approximately 3-fold greater than that observed on VERO cells; a difference found to be due to the differential expression of cell surface GlcNAc on HT29 and VERO cells. Further characterization of this activity using propidium iodine staining revealed that PAL induced cell cycle arrest at G2/M phase in a manner dependent on its ability to bind GlcNAc.

Runt related transcription factor 2 (RUNX2) is a key regulator of osteoblast differentiation. Several variations within the RUNX2 gene have been found to be associated with significant changes in BMD, which is a major risk factor for fracture. In this study we report that an 18 bp deletion within the polyalanine tract (17A>11A) of RUNX2 is significantly associated with fracture. Carriers of the 11A allele were found to be nearly twice as likely to have sustained fracture. Within the fracture category, there was a significant tendency of 11A carriers to present with fractures of distal radius and bones of intramembranous origin compared to bones of endochondral origin (p = 0.0001). In a population of random subjects, the 11A allele was associated with decreased levels of serum collagen cross links (CTx, p = 0.01), suggesting decreased bone turnover. The transactivation function of the 11A allele showed a minor quantitative decrease. Interestingly, we found no effect of the 11A allele on BMD at multiple skeletal sites. These findings suggest that the 11A allele is a biologically relevant polymorphism that influences serum CTx and confers enhanced fracture risk in a site-selective manner related to intramembranous bone ossification.

Item does not contain fulltext CMP-sialic acid transporter: We report an in-depth, multidisciplinary, structural study that has identified the amino acid residues intimately involved in CMP-sialic acid transporter (CST) substrate specificity. Our data provide a significant contribution towards a better understanding the structure-function relationship of this important family of transporters and the rational design of CST inhibitors.

RUNX2 is an essential transcription factor required for skeletal development and cartilage formation. Haploinsufficiency of RUNX2 leads to cleidocranial displaysia (CCD) a skeletal disorder characterised by gross dysgenesis of bones particularly those derived from intramembranous bone formation. A notable feature of the RUNX2 protein is the polyglutamine and polyalanine (23Q/17A) domain coded by a repeat sequence. Since none of the known mutations causing CCD characterised to date map in the glutamine repeat region, we hypothesised that Q-repeat mutations may be related to a more subtle bone phenotype. We screened subjects derived from four normal populations for Q-repeat variants. A total of 22 subjects were identified who were heterozygous for a wild type allele and a Q-repeat variant allele: (15Q, 16Q, 18Q and 30Q). Although not every subject had data for all measures, Q-repeat variants had a significant deficit in BMD with an average decrease of 0.7SD measured over 12 BMD-related parameters (p = 0.005). Femoral neck BMD was measured in all subjects (-0.6SD, p = 0.0007). The transactivation function of RUNX2 was determined for 16Q and 30Q alleles using a reporter gene assay. 16Q and 30Q alleles displayed significantly lower transactivation function compared to wild type (23Q). Our analysis has identified novel Q-repeat mutations that occur at a collective frequency of about 0.4%. These mutations significantly alter BMD and display impaired transactivation function, introducing a new class of functionally relevant RUNX2 mutants.

Background Real-time quantitative RT-PCR (qPCR) is a powerful technique capable of accurately quantitating mRNA expression levels over a large dynamic range. This makes qPCR the most widely used method for studying quantitative gene expression. An important aspect of qPCR is selecting appropriate controls or normalization factors to account for any differences in starting cDNA quantities between samples during expression studies. Here, we report on the selection of a concise set of housekeeper genes for the accurate normalization of quantitative gene expression data in differentiating osteoblasts, osteoclasts and macrophages. We implemented the use of geNorm, an algorithm that determines the suitability of genes to function as housekeepers by assessing expression stabilities. We evaluated the expression stabilities of 18S, ACTB, B2M, GAPDH, HMBS and HPRT1 genes. Findings Our analyses revealed that 18S and GAPDH were regulated during osteoblast differentiation and are not suitable for use as reference genes. The most stably expressed genes in osteoblasts were ACTB, HMBS and HPRT1 and their geometric average constitutes a suitable normalization factor upon which gene expression data can be normalized. In macrophages, 18S and GAPDH were the most variable genes while HMBS and B2M were the most stably expressed genes. The geometric average of HMBS and B2M expression levels forms a suitable normalization factor to account for potential differences in starting cDNA quantities during gene expression analysis in macrophages. The expression stabilities of the six candidate reference genes in osteoclasts were, on average, more variable than that observed in macrophages but slightly less variable than those seen in osteoblasts. The two most stably expressed genes in osteoclasts were HMBS and B2M and the genes displaying the greatest levels of variability were 18S and GAPDH. Notably, 18S and GAPDH were the two most variably expressed control genes in all three cell types. The geometric average of HMBS, B2M and ACTB creates an appropriate normalization factor for gene expression studies in osteoclasts. Conclusion We have identified concise sets of genes suitable to use as normalization factors for quantitative real-time RT-PCR gene expression studies in osteoblasts, osteoclasts and macrophages.

Rapid mineralization of cultured osteoblasts could be a useful characteristic in stem cell-mediated therapies for fracture and other orthopedic problems. Dimethyl sulfoxide (DMSO) is a small amphipathic solvent molecule capable of stimulating cell differentiation. We report that, in primary human osteoblasts, DMSO dose-dependently enhanced the expression of osteoblast differentiation markers alkaline phosphatase activity and extracellular matrix mineralization. Furthermore, similar DMSO-mediated mineralization enhancement was observed in primary osteoblast-like cells differentiated from mouse mesenchymal cells derived from fat, a promising source of starter cells for cell-based therapy. Using a convenient mouse pre-osteoblast model cell line MC3T3-E1, we further investigated this phenomenon showing that numerous osteoblast-expressed genes were elevated in response to DMSO treatment and correlated with enhanced mineralization. Myocyte enhancer factor 2c (Mef2c) was identified as the transcription factor most induced by DMSO, among the numerous DMSO-induced genes, suggesting a role for Mef2c in osteoblast gene regulation. Immunohistochemistry confirmed expression of Mef2c in osteoblast-like cells in mouse mandible, cortical, and trabecular bone. shRNAi-mediated Mef2c gene silencing resulted in defective osteoblast differentiation, decreased alkaline phosphatase activity, and matrix mineralization and knockdown of osteoblast specific gene expression, including osteocalcin and bone sialoprotein. A flow on knockdown of bone-specific transcription factors, Runx2 and osterix by shRNAi knockdown of Mef2c, suggests that Mef2c lies upstream of these two important factors in the cascade of gene expression in osteoblasts.

Loading of the rat ulna is an ideal model to examine stress fracture healing. The aim of this study was to undertake a detailed examination of the histology, histomorphometry and gene expression of the healing and remodelling process initiated by fatigue loading of the rat ulna. Ulnae were harvested 1, 2, 4, 6, 8, and 10 weeks following creation of a stress fracture. Stress fracture healing involved direct remodelling that progressed along the fracture line as well as woven bone proliferation at the site of the fracture. Histomorphometry demonstrated rapid progression of basic multicellular units from 1 to 4 weeks with significant slowing down of healing by 10 weeks after loading. Quantitative PCR was performed at 4 hours, 24 hours, 4 days, 7 days, and 14 days after loading. Gene expression was compared to an unloaded control group. At 4 hours after fracture, there was a marked 220-fold increase (P

Objective To determine a practical approach for deriving life expectancy estimates in Australian New South Wales local government areas which display a large diversity in population sizes. Design Population-based study utilising mortality and estimated residential population data. Setting 153 local government areas in New South Wales, Australia. Outcome measures Key performance measures of Chiang II, Silcocks, adjusted Chiang II and Bayesian random effects model methodologies of life expectancy estimation including agreement analysis of life expectancy estimates and comparison of estimate SEs. Results Chiang II and Silcocks methods produced almost identical life expectancy estimates across a large range of population sizes but calculation failures and excessively large SEs limited their use in small populations. A population of 25 000 or greater was required to estimate life expectancy with SE of 1 year or less using adjusted Chiang II (a composite of Chiang II and Silcocks methods). Data aggregation offered some remedy for extending the use of adjusted Chiang II in small populations but reduced estimate currency. A recently developed Bayesian random effects model utilising the correlation in mortality rates between genders, age groups and geographical areas markedly improved the precision of life expectancy estimates in small populations. Conclusions We propose a hybrid approach for the calculation of life expectancy using the Bayesian random effects model in populations of 25 000 or lower permitting the precise derivation of life expectancy in small populations. In populations above 25 000, we propose the use of adjusted Chiang II to guard against violations of spatial correlation, to benefit from a widely accepted method that is simpler to communicate to local health authorities and where its slight inferior performance compared with the Bayesian approach is of minor practical significance.