Emergency presentations for farm-related injuries in older adults residing in south-western Victoria, Australia.

INTRODUCTION: Farm workers are at high risk for injuries, and epidemiological data are needed to plan resource allocation. OBJECTIVE: This study identified regions with high farm-related injury rates in the Barwon South West region of Victoria, Australia, for residents aged ≥50 yr. DESIGN: Retrospective synthesis using electronic medical records of emergency presentations occurring during 2017-2019 inclusive for Local Government Areas (LGA) in the study region. For each LGA, age-standardised incidence rates (per 1000 population/year) were calculated. FINDINGS: For men and women combined, there were 31 218 emergency presentations for any injury, and 1150 (3.68%) of these were farm-related. The overall age-standardised rate for farm-related injury presentations was 2.6 (95% CI 2.4-2.7); men had a higher rate than women (4.1, 95% CI 3.9-4.4 versus 1.2, 95% CI 1.0-1.3, respectively). For individual LGAs, the highest rates of farm-related emergency presentations occurred in Moyne and Southern Grampians, both rural LGAs. Approximately two-thirds of farm-related injuries occurred during work activities (65.0%), and most individuals arrived at the hospital by transport classified as "other" (including private car, 83.3%). There were also several common injury causes identified: "other animal related injury" (20.2%), "cutting, piercing object" (19.5%), "fall ⟨1 m" (13.1%), and "struck by or collision with object" (12.5%). Few injuries were caused by machinery (1.7%) and these occurred mainly in the LGA of Moyne (65%). DISCUSSION AND CONCLUSION: This study provides data to inform future research and resource allocation for the prevention of farm-related injuries.

Rural suicide risk and physical ill health: A qualitative study of the Victorian Suicide Register, 2009-2015.

OBJECTIVE: To gain new insight into contextual factors shaping how physical ill health acts as a stressor in rural suicides-informing the development of appropriate targeted interventions. DESIGN: Retrospective cohort study. SETTING: Non-metropolitan Victoria, Australia. SAMPLE: 802 rural (non-metropolitan) suicide deaths between the years 2009 and 2015. MAIN OUTCOME MEASURE: Qualitative data from the Victorian Suicide Register relating to physical ill health of suicide decedents. RESULTS: Thematic analysis identified 4 themes: (a) 'living with physical ill health in a rural area' highlights both practical and cultural challenges associated with managing physical ill health within the context of life in a rural setting; (b) 'causes and experience of chronic pain' highlights rural risk factors for chronic pain, how pain was experienced and treatment managed; (c) 'when living an independent, contributing life is no longer possible' describes the debilitating physical and mental outcomes of chronic ill health and pain; and (d) 'the cumulative impact of physical ill health and other factors on suicide risk' highlights the complexity of risk factors-in combination with (or as a result of) physical ill health-that contribute to a suicide death. CONCLUSION: Identified themes suggest pathways to improve understanding and support for those experiencing physical ill health and associated suicide risk. These supports include contextually and culturally appropriate rural services to provide effective and necessary treatment, pain relief and mental health support; acknowledgement and response to a culture of alcohol misuse as an (ultimately ineffective) coping strategy; proactive psychosocial support mechanisms; and alternative approaches to support including consideration of innovative peer support models.

Agricultural health and medicine education-Engaging rural professionals to make a difference to farmers' lives.

OBJECTIVE: Despite continued higher rates of workplace injuries, earlier morbidity and mortality and challenging climatic environments, few formal programs focus on the health, well-being and safety of farmers. The agricultural health and medicine unit, developed in 2010, was designed to increase cultural competence and empower rural professionals to improve the health, well-being and safety outcomes of farming populations in Australia. This study aimed to understand the extent to which graduates (2010-2018) use the knowledge and skills gained in their current occupations and identify barriers and enablers faced in implementing them. DESIGN: Mixed-methods descriptive study. SETTING: Graduates were invited to complete an online survey. Following the survey, graduates participated in a phone interview until saturation was reached. PARTICIPANTS: Forty-one graduates completed the survey (31% response rate), and eleven interviews were conducted. INTERVENTIONS: Education in agricultural health and medicine. MAIN OUTCOME MEASURES: Graduates use of knowledge and skills gained from the course and the barriers and enablers they experienced in implementation. RESULTS: The most represented occupations were nursing, medicine and agriculture (farming). Of respondents, 76% agreed their ability to diagnose, treat or prevent agricultural occupational illness or injury had improved. Positively, 42% use course content professionally at least weekly. Fifty-one per cent experienced barriers in implementing their new knowledge, and little evidence of career advancement was observed. CONCLUSION: This study informs the continuous development of the agricultural health and medicine curriculum and highlights the importance of a collaborative and multidisciplinary approach to improving the health, well-being and safety of farming populations. Despite engaged graduates, the continued high workplace mortality, preventable non-communicable disease and challenging climatic conditions highlight the need for strategic prioritisation of farmers' health across health, agriculture and policy settings.

Antibacterial Spectrum of a Tetrazole-Based Reversible Inhibitor of Serine ß-Lactamases.

CTX-M is the most prevalent family of extended-spectrum ß-lactamases. We recently developed a tetrazole-derived noncovalent inhibitor of CTX-M-9. Here, we present the biochemical and microbiological activity of this inhibitor across a representative panel of serine ß-lactamases and Gram-negative bacteria. The compound displayed significant activity against all major subgroups of CTX-M, including CTX-M-15, while it exhibited some low-level inhibition of other serine ß-lactamases. Complex crystal structures with the CTX-M-14 S237A mutant and CTX-M-27 illustrate the binding contribution of specific active-site residues on the ß3 strand. In vitro pharmacokinetic studies revealed drug-like properties and positive prospects for further optimization. These studies suggest that tetrazole-based compounds can provide novel chemotypes for future serine ß-lactamase inhibitor discovery.

Evidence for Inhibition of Topoisomerase 1A by Gold(III) Macrocycles and Chelates Targeting Mycobacterium tuberculosis and Mycobacterium abscessus.

Mycobacterium tuberculosis and the fast-growing species Mycobacterium abscessus are two important human pathogens causing persistent pulmonary infections that are difficult to cure and require long treatment times. The emergence of drug-resistant M. tuberculosis strains and the high level of intrinsic resistance of M. abscessus call for novel drug scaffolds that effectively target both pathogens. In this study, we evaluated the activity of bis(pyrrolide-imine) gold(III) macrocycles and chelates, originally designed as DNA intercalators capable of targeting human topoisomerase types I and II (Topo1 and Topo2), against M. abscessus and M. tuberculosis We identified a total of 5 noncytotoxic compounds active against both mycobacterial pathogens under replicating in vitro conditions. We chose one of these hits, compound 14, for detailed analysis due to its potent bactericidal mode of inhibition and scalable synthesis. The clinical relevance of this compound was demonstrated by its ability to inhibit a panel of diverse M. tuberculosis and M. abscessus clinical isolates. Prompted by previous data suggesting that compound 14 may target topoisomerase/gyrase enzymes, we demonstrated that it lacked cross-resistance with fluoroquinolones, which target the M. tuberculosis gyrase. In vitro enzyme assays confirmed the potent activity of compound 14 against bacterial topoisomerase 1A (Topo1) enzymes but not gyrase. Novel scaffolds like compound 14 with potent, selective bactericidal activity against M. tuberculosis and M. abscessus that act on validated but underexploited targets like Topo1 represent a promising starting point for the development of novel therapeutics for infections by pathogenic mycobacteria.

The "PepSAVI-MS" Pipeline for Natural Product Bioactive Peptide Discovery.

The recent increase in extensively drug-resistant bacterial pathogens and the associated increase of morbidity and mortality demonstrate the immediate need for new antibiotic backbones with novel mechanisms of action. Here, we report the development of the PepSAVI-MS pipeline for bioactive peptide discovery. This highly versatile platform employs mass spectrometry and statistics to identify bioactive peptide targets from complex biological samples. We validate the use of this platform through the successful identification of known bioactive peptides from a botanical species, Viola odorata. Using this pipeline, we have widened the known antimicrobial spectrum for V. odorata cyclotides, including antibacterial activity of cycloviolacin O2 against A. baumannii. We further demonstrate the broad applicability of the platform through the identification of novel anticancer activities for cycloviolacins by their cytotoxicity against ovarian, breast, and prostate cancer cell lines.

Proteolytic processing of the neuronal ceroid lipofuscinosis related lysosomal protein CLN5.

CLN5 is a soluble lysosomal glycoprotein. Deficiency in CLN5 protein causes neuronal ceroid lipofuscinosis, an inherited neurodegenerative lysosomal storage disorder. The function of CLN5 and how it affects lysosome activity are unclear. We identified two forms of the CLN5 protein present in most of the cell lines studied. The molecular mass difference between these two forms is about 4kDa. The fibroblast cells derived from two CLN5 patients lack both forms. Using transient transfection, we showed one of these two forms is a proprotein and the other is a C-terminal cleaved mature form. Using cycloheximide chase analysis, we were able to demonstrate that the C-terminal processing occurs post-translationally. By treating cells with several pharmaceutical drugs to inhibit proteases, we showed that the C-terminal processing takes place in an acidic compartment and the protease involved is most likely a cysteine protease. This is further supported by overexpression of a CLN5 patient mutant D279N and a glycosylation mutant N401Q, showing that the C-terminal processing takes place beyond the endoplasmic reticulum, and can occur as early as from the trans Golgi network. Furthermore, we demonstrated that CLN5 is expressed in a variety of murine tissues.

Utilizing the Delphi method to develop parent and child surveys to understand exposure to farming hazards and attitudes toward farm safety.

Children on farms are at increased risk of injury. In Australia, children under 15 years consistently represent ~15% of all farm-related fatalities. This study aimed to develop parent and child surveys to gain a greater understanding of children's (5-14 years) exposure to occupational risk on farms by exploring their exposure to farm hazards, risk-taking behavior, their use and attitudes toward safety measures, and experience of farm-related injury. As farming communities are heterogeneous, a modified Delphi method was undertaken to ensure input from a diverse group. Seventeen experts participated in a three round process-the first two rounds required rating of proposed survey questions in an online questionnaire and the final round was an online discussion. Consensus was defined as 75% agreement or higher. This process resulted in 155 parent questions and 124 child questions reaching consensus to include. The modified Delphi method developed surveys that provide insight into the behaviors and attitudes of children (individuals) and their parents on farms (family) and will assist in informing how community, organizations and policy frameworks can improve child safety on farms. It will assist in identifying and understanding common farming exposures/behaviors of children and their parents to inform the development of targeted and culturally appropriate injury prevention strategies. As farming groups are heterogeneous, these survey scan be used on varying farming cohorts to identify their unique farming hazards and challenges. Child farm-related injuries are a problem globally and must be addressed; children are dependent on adults and communities to create safe environments for them.

High-Heat Days and Presentations to Emergency Departments in Regional Victoria, Australia.

Heat kills more Australians than any other natural disaster. Previous Australian research has identified increases in Emergency Department presentations in capital cities; however, little research has examined the effects of heat in rural/regional locations. This retrospective cohort study aimed to determine if Emergency Department (ED) presentations across the south-west region of Victoria, Australia, increased on high-heat days (1 February 2017 to 31 January 2020) using the Rural Acute Hospital Data Register (RAHDaR). The study also explored differences in presentations between farming towns and non-farming towns. High-heat days were defined as days over the 95th temperature percentile. International Statistical Classification of Diseases and Related Health Problems, Tenth Revision, Australian Modification (ICD-10-AM) codes associated with heat-related illness were identified from previous studies. As the region has a large agricultural sector, a framework was developed to identify towns estimated to have 70% or more of the population involved in farming. Overall, there were 61,631 presentations from individuals residing in the nine Local Government Areas. Of these presentations, 3064 (5.0%) were on days of high-heat, and 58,567 (95.0%) were of days of non-high-heat. Unlike previous metropolitan studies, ED presentations in rural south-west Victoria decrease on high-heat days. This decrease was more prominent in the farming cohort; a potential explanation for this may be behavioural adaption.

Child Farm-Related Injury in Australia: A Review of the Literature.

Children on farms have been identified as a population vulnerable to injury. This review seeks to identify child farm-related injury rates in Australia and to determine the key hazards and contributing risk factors. This critical review utilised the PRISMA guidelines for database searching. Research from the year 2000 onward was included as well as earlier seminal texts. Reference lists were searched, and the relevant research material was explored. Our primary focus was on Australian peer-reviewed literature with international and grey literature examples included. Evidence suggests that there is limited Australian research focusing on child farm-related injuries. Child representation in farm-related injuries in Australia has remained consistent over time, and the key hazards causing these injuries have remained the same for over 20 years. The factors contributing to child rates of farm injury described in the literature include child development and exposure to dangerous environments, the risk-taking culture, multi-generational farming families, lack of supervision, child labour and lack of regulation, limited targeted farm safety programs, underuse of safe play areas, financial priorities and poor understanding and operationalisation of the hierarchy of control. It is well known that children experience injury on farms, and the key hazards that cause this have been clearly identified. However, the level of exposure to hazards and the typical attitudes, behaviours and actions of children and their parents around the farm that contribute to chid injury remain unexplored.

Suicide in Rural Australia: Are Farming-Related Suicides Different?

Rural Australians experience a range of health inequities-including higher rates of suicide-when compared to the general population. This retrospective cohort study compares demographic characteristics and suicide death circumstances of farming- and non-farming-related suicides in rural Victoria with the aim of: (a) exploring the contributing factors to farming-related suicide in Australia's largest agricultural producing state; and (b) examining whether farming-related suicides differ from suicide in rural communities. Farming-related suicide deaths were more likely to: (a) be employed at the time of death (52.6% vs. 37.7%, OR = 1.84, 95% CIs 1.28-2.64); and, (b) have died through use of a firearm (30.1% vs. 8.7%, OR = 4.51, 95% CIs 2.97-6.92). However, farming-related suicides were less likely to (a) have a diagnosed mental illness (36.1% vs. 46.1%, OR=0.66, 95% CIs 0.46-0.96) and, (b) have received mental health support more than six weeks prior to death (39.8% vs. 50.0%, OR = 0.66, 95% CIs 0.46-0.95). A range of suicide prevention strategies need adopting across all segments of the rural population irrespective of farming status. However, data from farming-related suicides highlight the need for targeted firearm-related suicide prevention measures and appropriate, tailored and accessible support services to support health, well-being and safety for members of farming communities.

Autophagy-lysosome pathway alterations and alpha-synuclein up-regulation in the subtype of neuronal ceroid lipofuscinosis, CLN5 disease.

Neuronal ceroid lipofuscinoses (NCLs) are a group of inherited neurodegenerative lysosomal storage disorders. CLN5 deficiency causes a subtype of NCL, referred to as CLN5 disease. CLN5 is a soluble lysosomal protein with an unclear function in the cell. Increased levels of the autophagy marker protein LC3-II have been reported in several subtypes of NCLs. In this report, we examine whether autophagy is altered in CLN5 disease. We found that the basal level of LC3-II was elevated in both CLN5 disease patient fibroblasts and CLN5-deficient HeLa cells. Further analysis using tandem fluorescent mRFP-GFP-LC3 showed the autophagy flux was increased. We found the alpha-synuclein (α-syn) gene SNCA was highly up-regulated in CLN5 disease patient fibroblasts. The aggregated form of α-syn is well known for its role in the pathogenicity of Parkinson's disease. Higher α-syn protein levels confirmed the SNCA up-regulation in both patient cells and CLN5 knockdown HeLa cells. Furthermore, α-syn was localized to the vicinity of lysosomes in CLN5 deficient cells, indicating it may have a lysosome-related function. Intriguingly, knocking down SNCA reversed lysosomal perinuclear clustering caused by CLN5 deficiency. These results suggest α-syn may affect lysosomal clustering in non-neuronal cells, similar to its role in presynaptic vesicles in neurons.

Heteroaryl Phosphonates as Noncovalent Inhibitors of Both Serine- and Metallocarbapenemases.

Gram-negative pathogens expressing serine ß-lactamases (SBLs) and metallo-ß-lactamases (MBLs), especially those with carbapenemase activity, threaten the clinical utility of almost all ß-lactam antibiotics. Here we describe the discovery of a heteroaryl phosphonate scaffold that exhibits noncovalent cross-class inhibition of representative carbapenemases, specifically the SBL KPC-2 and the MBLs NDM-1 and VIM-2. The most potent lead, compound 16, exhibited low nM to low µM inhibition of KPC-2, NDM-1, and VIM-2. Compound 16 potentiated imipenem efficacy against resistant clinical and laboratory bacterial strains expressing carbapenemases while showing some cytotoxicity toward human HEK293T cells only at concentrations above 100 µg/mL. Complex structures with KPC-2, NDM-1, and VIM-2 demonstrate how these inhibitors achieve high binding affinity to both enzyme classes. These findings provide a structurally and mechanistically new scaffold for drug discovery targeting multidrug resistant Gram-negative pathogens and more generally highlight the active site features of carbapenemases that can be leveraged for lead discovery.

Physical Activity and Fundamental Motor Skill Performance of 5⁻10 Year Old Children in Three Different Playgrounds.

Playgrounds provide opportunities for children to engage in physical activity and develop their fundamental motor skills. The aim of this descriptive pilot study was to examine whether playground design facilitated different levels of physical activity and fundamental motor skills. Children aged 5 to 10 (n = 57) were recruited from three independent playgrounds located in Melbourne (Australia). Whilst playing, children wore accelerometers which measured time spent in physical activity and direct observations recorded fundamental motor skills and play equipment use. A general linear model with playground type as the predictor and adjusting for monitor wear-time identified whether mean time in physical activity was different for the three playgrounds. Frequencies and a one-way ANOVA assessed whether the observed mean number of fundamental motor skills varied between playgrounds. On average, 38.1% of time (12.0 min) was spent in moderate- vigorous-intensity physical activity. Children in the traditional playground (n = 16) engaged in more moderate-intensity physical activity (9.4 min) than children in the adventure playground (n = 21), (5.6 min) (p = 0.027). There were no significant associations with vigorous-intensity physical activity or fundamental motor skills between playgrounds. Children performed few fundamental motor skills but used a wider variety of equipment in the contemporary and adventure playgrounds. Playgrounds need to maximise opportunities for children to engage in physical activity and develop fundamental motor skills.

Identification of a Novel Polyamine Scaffold With Potent Efflux Pump Inhibition Activity Toward Multi-Drug Resistant Bacterial Pathogens.

We have previously reported the use of combinatorial chemistry to identify broad-spectrum antibacterial agents. Herein, we extend our analysis of this technology toward the discovery of anti-resistance molecules, focusing on efflux pump inhibitors. Using high-throughput screening against multi-drug resistant Pseudomonas aeruginosa, we identified a polyamine scaffold that demonstrated strong efflux pump inhibition without possessing antibacterial effects. We determined that these molecules were most effective with an amine functionality at R1 and benzene functionalities at R2 and R3. From a library of 188 compounds, we studied the properties of 5 lead agents in detail, observing a fivefold to eightfold decrease in the 90% effective concentration of tetracycline, chloramphenicol, and aztreonam toward P. aeruginosa isolates. Additionally, we determined that our molecules were not only active toward P. aeruginosa, but toward Acinetobacter baumannii and Staphylococcus aureus as well. The specificity of our molecules to efflux pump inhibition was confirmed using ethidium bromide accumulation assays, and in studies with strains that displayed varying abilities in their efflux potential. When assessing off target effects we observed no disruption of bacterial membrane polarity, no general toxicity toward mammalian cells, and no inhibition of calcium channel activity in human kidney cells. Finally, combination treatment with our lead agents engendered a marked increase in the bactericidal capacity of tetracycline, and significantly decreased viability within P. aeruginosa biofilms. As such, we report a unique polyamine scaffold that has strong potential for the future development of novel and broadly active efflux pump inhibitors targeting multi-drug resistant bacterial infections.