A foodborne outbreak of campylobacteriosis at a wedding - Melbourne, Australia, 2022.

Campylobacter is the most common bacterial cause of foodborne gastroenteritis in Australia; however, outbreaks caused by the pathogen are relatively uncommon. In March 2022, the Victorian Department of Health was notified of a gastrointestinal illness in 20 guests following attendance at a wedding reception. Two of these individuals were notified with laboratory-confirmed campylobacteriosis, and an investigation was undertaken to identify the source of the infection and implement strategies to prevent further illness. A case-control study was conducted to determine the likely source of infection. Cases were defined as attendees of the wedding reception, with onset of diarrhoea and/or abdominal cramping 1-10 days after attending the function. Controls were randomly selected from the remaining list of non-ill guests. Cases and controls were interviewed using a standardised, menu-based questionnaire. Food preparation processes were documented, and food samples collected. A total of 29 wedding guests met the case definition. Cases reported onset of illness 2-5 days following the wedding and major symptoms included abdominal cramping (100%), diarrhoea (90%), headache (79%), and fever (62%). Two cases were hospitalised, one with ongoing secondary neurological sequelae. Illness was significantly associated with consumption of a duck breast brioche canapé containing duck liver parfait (odds ratio = 2.85; 95% confidence interval: 1.03-7.86). No leftover food samples were available for testing. The investigation found that the duck canapé was the likely vehicle of infection. Consistent with the literature on Campylobacter transmission, it is likely that inadequate cooking of the duck liver for the parfait was the contributing factor that led to illness. This highlights the risks posed by undercooked poultry dishes, and shows that education of food handlers remains a priority.

The impact of the 21 November 2016 epidemic thunderstorm asthma event on general practice clinics in metropolitan Melbourne, Australia.

BACKGROUND AND OBJECTIVES: On 21 November 2016, parts of Victoria experienced a devastating epidemic thunderstorm asthma (ETSA) event. The aim of this study was to describe the epidemiology and burden of the 2016 ETSA event at MedicineInsight-registered general practices in the Melbourne metropolitan area in Victoria, Australia. METHOD: A cross-sectional study was conducted using patient record data from 21-23 November 2016. Codes were developed to identify all patients presenting to MedicineInsight-registered general practices with asthma during the 2016 ETSA event. RESULTS: During the event, there were 787 more asthma-related encounters to MedicineInsight general practices than expected, which represented a 7.1-fold increase (605% increase). Estimates suggest that there were between approximately 8940 and 13,689 more asthma-related encounters than expected across metropolitan Melbourne. DISCUSSION: General practices were significantly affected by the 2016 ETSA event. This work highlights the important part that general practices play in responding to ETSA events and the need for these practices to be prepared to respond.

FNTB Promoter Polymorphisms Are Independent Predictors of Survival in Patients with Triple Negative Breast Cancer.

In breast cancer, the promising efficacy of farnesyltransferase inhibitors (FTIs) in preclinical studies is in contrast to only limited effects in clinical Phase II-III trials. The objective of this study was to explore the clinical relevance of farnesyltransferase ß-subunit (FNTB) single nucleotide promoter polymorphisms (FNTB-173 6G > 5G (rs3215788), -609 G > C (rs11623866) and -179 T > A (rs192403314)) in early breast cancer. FNTB genotyping was performed by pyrosequencing in 797 patients from a prospective multicentre observational PiA trial (NCT01592825). In the total cohort, the FNTB-173 6G > 5G polymorphism was an independent predictor of RFI (HR = 0.568; 95% CI = 0.339-0.949, p = 0.031), OS (HR = 0.629; 95% CI = 0.403-0.980, p = 0.040) and BCSS (HR = 0.433; 95% CI = 0.213-0.882; p = 0.021), whereas the FNTB-609 G > C polymorphism was an independent predictor of RFI (HR = 0.453; 95% CI = 0.226-0.910, p = 0.026) and BCSS (HR = 0.227; 95% CI = 0.075-0.687, p = 0.009). Subtype analysis revealed the independent prognostic relevance of FNTB promoter polymorphisms, particularly in TNBC but not in luminal or HER2-positive intrinsic subtypes. Finally, we used electrophoretic mobility shift assays (EMSAs) to confirm in vitro that the polymorphism FNTB-173 6G > 5G resulted in the differential binding of nuclear proteins from five different breast cancer cell lines. This is the first study on breast cancer suggesting that FNTB promoter polymorphisms (i) are independent prognostic biomarkers, particularly in patients with early TNBC, and (ii) could modulate FNTB's transcriptional activity.

Are convergence lines associated with high asthma presentation days? A case-control study in Melbourne, Australia.

BACKGROUND: In epidemic thunderstorm asthma (ETSA) events a large number of people develop asthma symptoms over a short period of time. This is thought to occur because of a unique combination of high amounts of pollen and certain meteorological conditions. However, the exact cause and mechanism of epidemic thunderstorm asthma remains unclear. OBJECTIVES: The objective of this study was to test the hypothesis that convergence lines may be a causative factor in ETSA events, by investigating whether convergence line weather events are associated with the occurrence of high asthma presentations days during the Victorian grass pollen season (October-December). METHODS: A case control method was used. All public hospitals within 75 km of the Melbourne weather radar were included, and data were taken from 2009 to 2017 during the Victorian grass pollen season. Cases hospital days were hospitals with a high number of asthma presentations within a 24-h period, and controls were hospitals with an expected number of asthma presentations. Exposure was defined as geographical proximity of a convergence line to the hospital case or control. RESULTS: Eighty-one case hospital days and 157 hospital day controls were included in the study. The odds of exposure to a convergence line were significantly higher for cases than for controls at all exposure distances. At 4 km, 80 of the 81 cases had been exposed to a convergence line. CONCLUSION: Convergence lines appear to be a necessary, but not sufficient, element in the cause of epidemic thunderstorm asthma. This is the first study to show a clear link between epidemic thunderstorm asthma and convergence lines.

The forecasting of dynamical Ross River virus outbreaks: Victoria, Australia.

Ross River virus (RRV) is Australia's most epidemiologically important mosquito-borne disease. During RRV epidemics in the State of Victoria (such as 2010/11 and 2016/17) notifications can account for up to 30% of national RRV notifications. However, little is known about factors which can forecast RRV transmission in Victoria. We aimed to understand factors associated with RRV transmission in epidemiologically important regions of Victoria and establish an early warning forecast system. We developed negative binomial regression models to forecast human RRV notifications across 11 Local Government Areas (LGAs) using climatic, environmental, and oceanographic variables. Data were collected from July 2008 to June 2018. Data from July 2008 to June 2012 were used as a training data set, while July 2012 to June 2018 were used as a testing data set. Evapotranspiration and precipitation were found to be common factors for forecasting RRV notifications across sites. Several site-specific factors were also important in forecasting RRV notifications which varied between LGA. From the 11 LGAs examined, nine experienced an outbreak in 2011/12 of which the models for these sites were a good fit. All 11 LGAs experienced an outbreak in 2016/17, however only six LGAs could predict the outbreak using the same model. We document similarities and differences in factors useful for forecasting RRV notifications across Victoria and demonstrate that readily available and inexpensive climate and environmental data can be used to predict epidemic periods in some areas. Furthermore, we highlight in certain regions the complexity of RRV transmission where additional epidemiological information is needed to accurately predict RRV activity. Our findings have been applied to produce a Ross River virus Outbreak Surveillance System (ROSS) to aid in public health decision making in Victoria.

A Qualitative Evaluation of a Decision Support System for District-Level Disease Surveillance in Sierra Leone.

A decision support system for district-level disease surveillance was piloted with the Port Loko District Health Management Team in Sierra Leone. Through a qualitative evaluation, the study explores the impact of the system on disease surveillance workflows. Results indicate that the system aided decision making for operational tasks, and reduced the time taken to analyze and report surveillance data. In addition, the study discusses the challenges of deploying a pilot system during the Ebola recovery in Sierra Leone, and proposes a high-level architecture for a modular, interoperable decision support system for disease surveillance for public health decision makers in low-resource health systems.

Sulfur K-Edge XAS Studies of the Effect of DNA Binding on the [Fe4S4] Site in EndoIII and MutY.

S K-edge X-ray absorption spectroscopy (XAS) was used to study the [Fe4S4] clusters in the DNA repair glycosylases EndoIII and MutY to evaluate the effects of DNA binding and solvation on Fe-S bond covalencies (i.e., the amount of S 3p character mixed into the Fe 3d valence orbitals). Increased covalencies in both iron-thiolate and iron-sulfide bonds would stabilize the oxidized state of the [Fe4S4] clusters. The results are compared to those on previously studied [Fe4S4] model complexes, ferredoxin (Fd), and to new data on high-potential iron-sulfur protein (HiPIP). A limited decrease in covalency is observed upon removal of solvent water from EndoIII and MutY, opposite to the significant increase observed for Fd, where the [Fe4S4] cluster is solvent exposed. Importantly, in EndoIII and MutY, a large increase in covalency is observed upon DNA binding, which is due to the effect of its negative charge on the iron-sulfur bonds. In EndoIII, this change in covalency can be quantified and makes a significant contribution to the observed decrease in reduction potential found experimentally in DNA repair proteins, enabling their HiPIP-like redox behavior.

Electrochemistry of the [4Fe4S] Cluster in Base Excision Repair Proteins: Tuning the Redox Potential with DNA.

Escherichia coli endonuclease III (EndoIII) and MutY are DNA glycosylases that contain [4Fe4S] clusters and that serve to maintain the integrity of the genome after oxidative stress. Electrochemical studies on highly oriented pyrolytic graphite (HOPG) revealed that DNA binding by EndoIII leads to a large negative shift in the midpoint potential of the cluster, consistent with stabilization of the oxidized [4Fe4S]3+ form. However, the smooth, hydrophobic HOPG surface is nonideal for working with proteins in the absence of DNA. In this work, we use thin film voltammetry on a pyrolytic graphite edge electrode to overcome these limitations. Improved adsorption leads to substantial signals for both EndoIII and MutY in the absence of DNA, and a large negative potential shift is retained with DNA present. In contrast, the EndoIII mutants E200K, Y205H, and K208E, which provide electrostatic perturbations in the vicinity of the cluster, all show DNA-free potentials within error of wild type; similarly, the presence of negatively charged poly-l-glutamate does not lead to a significant potential shift. Overall, binding to the DNA polyanion is the dominant effect in tuning the redox potential of the [4Fe4S] cluster, helping to explain why all DNA-binding proteins with [4Fe4S] clusters studied to date have similar DNA-bound potentials.

Design and evaluation of a web-based decision support tool for district-level disease surveillance in a low-resource setting.

During the 2014 West African Ebola Virus outbreak it became apparent that the initial response to the outbreak was hampered by limitations in the collection, aggregation, analysis and use of data for intervention planning. As part of the post-Ebola recovery phase, IBM Research Africa partnered with the Port Loko District Health Management Team (DHMT) in Sierra Leone and GOAL Global, to design, implement and deploy a web-based decision support tool for district-level disease surveillance. This paper discusses the design process and the functionality of the first version of the system. The paper presents evaluation results prior to a pilot deployment and identifies features for future iterations. A qualitative assessment of the tool prior to pilot deployment indicates that it improves the timeliness and ease of using data for making decisions at the DHMT level.

Characterization of the DNA-Mediated Oxidation of Dps, A Bacterial Ferritin.

Dps proteins are bacterial ferritins that protect DNA from oxidative stress and have been implicated in bacterial survival and virulence. In addition to direct oxidation of the Dps iron sites by diffusing oxidants, oxidation from a distance via DNA charge transport (CT), where electrons and electron holes are rapidly transported through the base-pair π-stack, could represent an efficient DNA protection mechanism utilized by Dps. Here, we spectroscopically characterize the DNA-mediated oxidation of ferrous iron-loaded Dps. X-band EPR was used to monitor the oxidation of DNA-bound Dps after DNA photooxidation using an intercalating ruthenium photooxidant and the flash-quench technique. Upon irradiation with poly(dGdC)2, a signal arises with g = 4.3, consistent with the formation of mononuclear high-spin Fe(III) sites of low symmetry, the expected oxidation product of Dps with one iron bound at each ferroxidase site. When poly(dGdC)2 is substituted with poly(dAdT)2, the yield of Dps oxidation is decreased significantly, consistent with guanine radical intermediates facilitating Dps oxidation. We have also explored possible protein electron transfer (ET) intermediates in the DNA-mediated oxidation of ferrous iron-loaded Dps. Dps proteins contain a conserved tryptophan residue in close proximity to the iron-binding ferroxidase site (W52 in E. coli Dps). In EPR studies of the oxidation of ferrous iron-loaded Dps following DNA photooxidation, a W52A Dps mutant was significantly deficient compared to WT Dps in forming the characteristic EPR signal at g = 4.3, consistent with W52 acting as an ET hopping intermediate. This effect is mirrored in vivo in E. coli survival in response to hydrogen peroxide, where mutation of W52 leads to decreased survival under oxidative stress.

DNA Charge Transport: from Chemical Principles to the Cell.

The DNA double helix has captured the imagination of many, bringing it to the forefront of biological research. DNA has unique features that extend our interest into areas of chemistry, physics, material science, and engineering. Our laboratory has focused on studies of DNA charge transport (CT), wherein charges can efficiently travel long molecular distances through the DNA helix while maintaining an exquisite sensitivity to base pair π-stacking. Because DNA CT chemistry reports on the integrity of the DNA duplex, this property may be exploited to develop electrochemical devices to detect DNA lesions and DNA-binding proteins. Furthermore, studies now indicate that DNA CT may also be used in the cell by, for example, DNA repair proteins, as a cellular diagnostic, in order to scan the genome to localize efficiently to damage sites. In this review, we describe this evolution of DNA CT chemistry from the discovery of fundamental chemical principles to applications in diagnostic strategies and possible roles in biology.

Kinetic results for mutations of conserved residues H304 and R309 of human sulfite oxidase point to mechanistic complexities.

Several point mutations in the gene of human sulfite oxidase (hSO) result in isolated sulfite oxidase deficiency, an inherited metabolic disorder. Three conserved residues (H304, R309, K322) are hydrogen bonded to the phosphate group of the molybdenum cofactor, and the R309H and K322R mutations are responsible for isolated sulfite oxidase deficiency. The kinetic effects of the K322R mutation have been previously reported (Rajapakshe et al., Chem. Biodiversity, 2012, 9, 1621-1634); here we investigate several mutants of H304 and R309 by steady-state kinetics, laser flash photolysis studies of intramolecular electron transfer (IET), and spectroelectrochemistry. An unexpected result is that all of the mutants show decreased rates of IET but increased steady-state rates of catalysis. However, in all cases the rate of IET is greater than the overall turnover rate, showing that IET is not the rate determining step for any of the mutations.

DNA protection by the bacterial ferritin Dps via DNA charge transport.

Dps proteins, bacterial mini-ferritins that protect DNA from oxidative stress, are implicated in the survival and virulence of pathogenic bacteria. Here we examine the mechanism of E. coli Dps protection of DNA, specifically whether this DNA-binding protein can utilize DNA charge transport through the base pair π-stack to protect the genome from a distance. An intercalating ruthenium photooxidant was employed to generate DNA damage localized to guanine repeats, the sites of lowest potential in DNA. We find that Dps loaded with ferrous iron, in contrast to Apo-Dps and ferric iron-loaded Dps, significantly attenuates the yield of oxidative DNA damage. These data demonstrate that ferrous iron-loaded Dps is selectively oxidized to fill guanine radical holes, thereby restoring the integrity of the DNA. Luminescence studies indicate no direct interaction between the ruthenium photooxidant and Dps, supporting the DNA-mediated oxidation of ferrous iron-loaded Dps. Thus DNA charge transport may be a mechanism by which Dps efficiently protects the genome of pathogenic bacteria from a distance.

Multiplexed electrochemistry of DNA-bound metalloproteins.

Here we describe a multiplexed electrochemical characterization of DNA-bound proteins containing [4Fe-4S] clusters. DNA-modified electrodes have become an essential tool for the characterization of the redox chemistry of DNA repair proteins containing redox cofactors, and multiplexing offers a means to probe different complex samples and substrates in parallel to elucidate this chemistry. Multiplexed analysis of endonuclease III (EndoIII), a DNA repair protein containing a [4Fe-4S] cluster known to be accessible via DNA-mediated charge transport, shows subtle differences in the electrochemical behavior as a function of DNA morphology. The peak splitting, signal broadness, sensitivity to π-stack perturbations, and kinetics were all characterized for the DNA-bound reduction of EndoIII on both closely and loosely packed DNA films. DNA-bound EndoIII is seen to have two different electron transfer pathways for reduction, either through the DNA base stack or through direct surface reduction; closely packed DNA films, where the protein has limited surface accessibility, produce electrochemical signals reflecting electron transfer that is DNA-mediated. Multiplexing furthermore permits the comparison of the electrochemistry of EndoIII mutants, including a new family of mutations altering the electrostatics surrounding the [4Fe-4S] cluster. While little change in the midpoint potential was found for this family of mutants, significant variations in the efficiency of DNA-mediated electron transfer were apparent. On the basis of the stability of these proteins, examined by circular dichroism, we propose that the electron transfer pathway can be perturbed not only by the removal of aromatic residues but also through changes in solvation near the cluster.

Probing the role of a conserved salt bridge in the intramolecular electron transfer kinetics of human sulfite oxidase.

Sulfite oxidase (SO) is a vital metabolic enzyme that catalyzes the oxidation of toxic sulfite to sulfate. The proposed mechanism of this molybdenum cofactor dependent enzyme involves two one-electron intramolecular electron transfer (IET) steps from the molybdenum center to the iron of the b 5-type heme and two one-electron intermolecular electron transfer steps from the heme to cytochrome c. This work focuses on how the electrostatic interaction between two conserved amino acid residues, R472 and D342, in human SO (hSO) affects catalysis. The hSO variants R472M, R472Q, R472K, R472D, and D342K were created to probe the effect of the position of the salt bridge charges, along with the interaction between these two residues. With the exception of R472K, these variants all showed a significant decrease in their IET rate constants, k et, relative to wild-type hSO, indicating that the salt bridge between residues 472 and 342 is important for rapid IET. Surprisingly, however, except for R472K and R472D, all of the variants show k cat values higher than their corresponding k et values. The turnover number for R472D is about the same as k et, which suggests that the change in this variant is rate-limiting in catalysis. Direct spectroelectrochemical determination of the Fe(III/II) reduction potentials of the heme and calculation of the Mo(VI/V) potentials revealed that all of the variants affected the redox potentials of both metal centers, probably due to changes in their environments. Thus, the position of the positive charge of R472 and that of the negative charge of D342 are both important in hSO, and changing either the position or the nature of these charges perturbs IET and catalysis.

Near-vision impairment and unresolved vision problems in Indigenous Australian adults.

BACKGROUND: To describe near-vision impairment, self-reported unresolved vision problems and barriers to having near-vision correction in Indigenous Australians. DESIGN: A nationwide population-based study designed to determine the causes and prevalence of vision loss and utilization of eye care services. PARTICIPANTS: Indigenous Australians aged ≥40 years. METHODS: Using a multistage random cluster sampling methodology, 30 geographical areas stratified by remoteness were selected to obtain a representation of Indigenous Australians. Visual acuity was conducted using a standard E chart. A questionnaire collected data on eye health, eye care service utilization and vision-related quality of life. MAIN OUTCOME MEASURES: Near-vision impairment defined as presenting binocular near visual acuity

Identity of the exchangeable sulfur-containing ligand at the Mo(V) center of R160Q human sulfite oxidase.

In our previous study of the fatal R160Q mutant of human sulfite oxidase (hSO) at low pH (Astashkin et al. J. Am. Chem. Soc.2008, 130, 8471-8480), a new Mo(V) species, denoted "species 1", was observed at low pH values. Species 1 was ascribed to a six-coordinate Mo(V) center with an exchangeable terminal oxo ligand and an equatorial sulfate group on the basis of pulsed EPR spectroscopy and (33)S and (17)O labeling. Here we report new results for species 1 of R160Q, based on substitution of the sulfur-containing ligand by a phosphate group, pulsed EPR spectroscopy in K(a)- and W-bands, and extensive density functional theory (DFT) calculations applied to large, more realistic molecular models of the enzyme active site. The combined results unambiguously show that species 1 has an equatorial sulfite as the only exchangeable ligand. The two types of (17)O signals that are observed arise from the coordinated and remote oxygen atoms of the sulfite ligand. A typical five-coordinate Mo(V) site is compatible with the observed and calculated EPR parameters.

The Impact of Knowledge and Attitudes on Access to Eye-Care Services in Cambodia.

PURPOSE: To investigate how knowledge and attitudes influence the access to eye-care services in Takeo Province, Cambodia. DESIGN: A cross-sectional survey (n=600). METHODS: 30 villages were randomly selected. Groups included: >50 years, 30-49 years, and parents with children <5 years. A newly developed Knowledge, Attitude and Practice in Eye Health (KAP-EH) questionnaire about knowledge and treatment of eye diseases, practices and attitudes to accessing services was used to interview respondents. Descriptive analysis, including Chi square and logistic regression tested for associations with sub-groups of gender, age group, education and self-reported type of disability. RESULTS: The proportion of respondents who reported having knowledge of specific eye conditions ranged from 97% for eye injury, to 8% for diabetic eye disease. While 509 (85%) people reported knowledge of cataract, 47% did not know how cataract was treated and only 19% listed surgery. The older group (66.5%) were least informed about cataract (p= 0.001) compared to other groups, and were least likely to believe that some blindness could be prevented (p < 0.001). Women (55%) were more likely than men (46%) (p=0 .003) to report that a child with blindness could attend school, as did people without a disability compared to those with a disability (58% vs 34%) (p < 0.001). CONCLUSIONS: The knowledge about cataract and refractive error and what to do to resolve the problems was low among this population and this study suggests that poor knowledge of eye diseases might contribute to the occurrence of un-operated cataract and uncorrected refractive error.

In the heart or in the head: relationship and cognitive influences on sexual risk among young couples.

Few studies examine how relationship factors influence sexual risk. We investigated gender differences of the influence of relationship functioning on sexual risk behavior and sexually transmitted infections (STIs) among 296 young pregnant couples. Compared to women, men were twice as likely to have a concurrent partner, were less likely to intend to be monogamous in the future, had less HIV/STI knowledge and more perceived risk for HIV/STI, negative attitudes and efficacy toward condom use (all P < 0.05). For men, poor relationship functioning related to less intentions to be monogamous, more partner concurrency, worse sexual communication, and more condom use. For women, poor relationship functioning related to worse sexual communication and less condom use. In addition, women who had good relationship functioning and women who had partners with poor relationship functioning were more likely to have an STI. These results demonstrate the need to include heterosexual men and relationship factors in HIV prevention.