The chaperone Chs7 forms a stable complex with Chs3 and promotes its activity at the cell surface.

The polytopic yeast protein Chs3 (chitin synthase III) relies on a dedicated membrane-localized chaperone, Chs7, for its folding and expression at the cell surface. In the absence of Chs7, Chs3 forms high molecular weight aggregates and is retained in the endoplasmic reticulum (ER). Chs7 was reported to be an ER resident protein, but its role in Chs3 folding and transport was not well characterized. Here, we show that Chs7 itself exits the ER and localizes with Chs3 at the bud neck and intracellular compartments. We identified mutations in the Chs7 C-terminal cytosolic domain that do not affect its chaperone function, but cause it to dissociate from Chs3 at a post-ER transport step. Mutations that prevent the continued association of Chs7 with Chs3 do not block delivery of Chs3 to the cell surface, but dramatically reduce its catalytic activity. This suggests that Chs7 engages in functionally distinct interactions with Chs3 to first promote its folding and ER exit, and subsequently to regulate its activity at the plasma membrane.

Assessment of the first 5 years of pharmacist-administered vaccinations in Australia: learnings to inform expansion of services.

Objectives and importance of the study: Pharmacist-administered vaccination has expanded in Australia but has not been comprehensively assessed. We aimed to assess the pharmacists' role in vaccination in Australia before and during the first year of the COVID-19 pandemic, as well as the completeness of data on pharmacist-administered immunisations. STUDY TYPE: Cross-sectional study. METHODS: We analysed data on pharmacist-administered vaccinations that were reported to the Australian Immunisation Register (AIR) between 2016 and 2019, categorised by gender, jurisdiction, age group and vaccine type. We conducted a national survey of community pharmacists providing vaccination services during June and July 2020 to understand how pharmacists record and report vaccinations to the AIR. We assessed data completeness by comparing the number of vaccinations reported by surveyed pharmacists to the number recorded on the AIR. RESULTS: 576 780 pharmacist-administered vaccinations were recorded on the AIR between 2016 and 2019, of which 94.7% were influenza vaccines. The proportion of vaccinations given by pharmacists increased each year, from <0.001% in 2016 to 2.7% in 2019. Between 2017 and 2019, rates of pharmacist-administered vaccinations were highest among people aged 60-64 years (2046 per 100 000 people) and those living in regional areas (1074 per 100 000 people). Among 243 survey respondents, 57.8% (126/223) reported vaccinations to the AIR automatically via software, 27.8% (62/223) manually entered data and 13.5% (30/223) used both methods. Of the 87 665 vaccination encounters recorded by 121 respondents, 82.2% (72 045/87 665) were recorded on the AIR. There were more AIR-recorded encounters from those who reported automatically via software (84.8% [49 309/58 134]) than from those who manually entered data (68.3% [12 127/17 746]). CONCLUSIONS: Pharmacists have an increasing role in providing vaccination services in Australia, with great potential to improve coverage among adults and populations in regional locations. Measures introduced during the COVID-19 pandemic may have increased the uptake of electronic methods of recording and reporting data, which can improve data completeness. Our results provide an assessment of the first 5 years of pharmacist vaccination services in Australia, against which future evaluations of the impacts of policy changes during the COVID-19 pandemic can be compared.

How accurately does the Australian Immunisation Register identify children overdue for vaccine doses? A national cross-sectional study.

Abstract: The accuracy of data recorded in the Australian Immunisation Register (AIR) is important for assessment of population-level vaccine coverage but has not been assessed nationally since 2001. We undertook a cross-sectional study in five states in 2017 using standard criteria to validate AIR records classified as three months overdue for any vaccine at 12, 24 and 48 months. Of 2,000 records selected for audit, 905 were assessable, of which 124 (14%) were misclassified as overdue (errors). Among 563 general practice (GP) records, 91 (16.1%) were errors. Compared with Victoria (1/99; 1%), errors were significantly higher in Western Australia (11/106; 10.4%), Queensland (13/104; 12.5%), South Australia (23/110; 20.9%) and New South Wales (43/144; 29.9%); p < 0.01 for all. Among 165 council and community health centre providers, the overall error rate (17; 10.3%) was non-significantly lower than for GP providers, with an odds ratio (OR) of 0.6 and a 95% confidence interval (95% CI) of 0.3-1.1, and did not differ between states. Records were transmitted to the AIR by paper-based methods in 13 cases, with significantly higher error rates (7/13; 54%) than for practice management software (77/630; 12.2%); OR 9.8 (95% CI 2.8-36.4) or the AIR secure site (23/87; 26.4%); OR 2.6 (95% CI 1.4-4.5). Accuracy is increasingly important, with mandatory reporting to the AIR for all National Immunisation Program vaccines from July 2021, and best achieved by uniform use of practice management software.

Quantitative high-content imaging identifies novel regulators of Neo1 trafficking at endosomes.

P4-ATPases are a family of putative phospholipid flippases that regulate lipid membrane asymmetry, which is important for vesicle formation. Two yeast flippases, Drs2 and Neo1, have nonredundant functions in the recycling of the synaptobrevin-like v-SNARE Snc1 from early endosomes. Drs2 activity is needed to form vesicles and regulate its own trafficking, suggesting that flippase activity and localization are linked. However, the role of Neo1 in endosomal recycling is not well characterized. To identify novel regulators of Neo1 trafficking and activity at endosomes, we first identified mutants with impaired recycling of a Snc1-based reporter and subsequently used high-content microscopy to classify these mutants based on the localization of Neo1 or its binding partners, Mon2 and Dop1. This analysis identified a role for Arl1 in stabilizing the Mon2/Dop1 complex and uncovered a new function for Vps13 in early endosome recycling and Neo1 localization. We further showed that the cargo-selective sorting nexin Snx3 is required for Neo1 trafficking and identified an Snx3 sorting motif in the Neo1 N-terminus. Of importance, the Snx3-dependent sorting of Neo1 was required for the correct sorting of another Snx3 cargo protein, suggesting that the incorporation of Neo1 into recycling tubules may influence their formation.

Large-scale analysis of membrane transport in yeast using invertase reporters.

Transport of membrane proteins between cellular organelles requires the concerted action of many regulatory factors, which aid in cargo recognition and vesicle formation, targeting, and fusion. The yeast Saccharomyces cerevisiae is a useful model system for studying such regulators, due to the availability of genome-wide mutant collections and reporter proteins that provide sensitive biochemical readouts of individual transport pathways. Here, we describe an enzymatic invertase assay for evaluating endocytic recycling using a chimeric GFP-Snc1-Suc2 reporter. Cell surface levels of this reporter can be measured by a colorimetric assay that monitors sucrose hydrolysis at the plasma membrane, using two different methods. The first is a semiquantitative agar overlay assay followed by image densitometry that is suitable for high-throughput screening of arrayed yeast colonies. In the second, more quantitative assay, an enzymatic solution is added to yeast cultures in a multi-well plate and the absorbance is assessed by a plate reader. Furthermore, the modular nature of the chimeric reporter allows alternate transport signals to be introduced, thereby expanding the range of transport pathways that can be evaluated by this method. Together these techniques can be used to explore the function of genes involved in a variety of cellular trafficking pathways.

Helping medical specialists working in rural and remote Australia deal with professional isolation: the Support Scheme for Rural Specialists.

There are well documented geographical, financial, social and professional barriers to continuing professional development (CPD) and peer support for rural medical practitioners, which significantly influence the recruitment and retention of health care professionals in rural areas. The Support Scheme for Rural Specialists (SSRS) provides a coordinated and collaborative framework to support the CPD and peer-support needs of medical specialists practising in rural and remote Australia. Since 2002, more than 80 CPD projects have been implemented by specialist medical colleges under the auspices of the SSRS. Projects have provided educational up-skilling or support for rural-specific clinical practice improvement initiatives aimed at strengthening clinician competence and capability, and workforce retention.

An educational workshop program for rural practitioners to encourage best practice for delivery of systemic adjuvant therapy.

BACKGROUND: The Rural Systemic Adjuvant Therapy Project was initiated to encourage best practice in the treatment of women from rural areas who have breast cancer. METHOD: We developed an educational program, piloted it and conducted it in 5 regions. In a pre-evaluation/post-evaluation, we assessed participants' perceived knowledge about systemic adjuvant therapy. RESULTS: A statistically significant increase occurred in participants' reported knowledge about all program topics. Improved communication links with the local or visiting medical oncologist were planned. CONCLUSION: The workshop program was found to be a successful tool for delivering evidence-based information about the use of systemic adjuvant therapy.