'Closing the gap': Evaluating the success of non-mandatory strategies for influenza vaccination of Victorian healthcare workers.

INTRODUCTION: While annual influenza vaccination of healthcare workers (HCWs) is recommended, uptake is often suboptimal. We sought to evaluate influenza vaccination uptake by HCWs in Victorian public healthcare facilities, where non-mandatory programs are used. METHODS: All participating facilities completed an annual survey (2014-2019) recording HCW influenza vaccination status. Uptake in high-risk departments (emergency and intensive care units) was evaluated for the 2019 season. RESULTS: The proportion of vaccinated HCWs increased annually, from 72.2% (2014) to 87.7% (2019), with pre-set targets generally achieved. In 2019, 110,324 HCWs in 107 facilities were vaccinated (87.7%). Of those without documented vaccination, 7591 (6.0%) declined and 7906 (6.3%) had unknown status. Uptake was higher in high-risk departments (91.4%). CONCLUSION: Increasing annual influenza vaccination uptake by HCWs in Victorian public healthcare facilities has been achieved in the context of performance monitoring targets. Small proportions declined or had unknown status. Future policies should focus on these HCWs.

Development and evaluation of a website for surveillance of healthcare-associated urinary tract infections in Australia.

Phase II of the Surveillance to Reduce Urinary Tract Infections project piloted a website for point prevalence surveys of healthcare-associated (HAUTI) and catheter-associated urinary tract infection in Australian hospitals and aged care homes. This report describes development and evaluation of the website for online data collection. Evaluation findings from 38 data collectors indicated that most respondents found website registration and web form use easy (N = 22; 58% and N = 16; 43%, respectively). The need for improved computer literacy skills and automated data systems were highlighted. This study demonstrated a novel approach for Australian HAUTI data collection; however, refinements are needed before national roll-out.

Validation of healthcare-associated infection surveillance in smaller Australian hospitals.

A validation study was conducted in smaller (<100 acute beds) Victorian hospitals to evaluate case detection for Staphylococcus aureus bloodstream (SAB), meticillin-resistant S. aureus (MRSA) and vancomycin-resistant enterococci (VRE) infections. Overall, 142 infections were identified in 20 hospitals. For routine surveillance of SAB events, sensitivity was 74.4% and specificity was 100.0%. For MRSA infections, sensitivity was 47.5% and specificity was 90.9%. All confirmed VRE infections were reported correctly. Of unreported SAB and MRSA infections, 80% (N = 16) and 83.9% (N = 26) were community-associated infections, respectively. Future programme refinements include targeted education to ensure appropriate application of case definitions, particularly those including community onset.

Fluorescent Labeling of Proteins in Whole Cell Extracts for Single-Molecule Imaging.

Cellular machines such as the spliceosome and ribosome can be composed of dozens of individual proteins and nucleic acids. Given this complexity, it is not surprising that many cellular activities have not yet been biochemically reconstituted. Such processes are often studied in vitro in whole cell or fractionated lysates. This presents a challenge for obtaining detailed biochemical information when the components being investigated may be only a minor component of the extract and unrelated processes may interfere with the assay. Single-molecule fluorescence microscopy methods allow particular biomolecules to be analyzed even in the complex milieu of a cell extract. This is due to the use of bright fluorophores that emit light at wavelengths at which few cellular components fluoresce, and the development of chemical biology tools for attaching these fluorophores to specific cellular proteins. Here, we describe a protocol for fluorescent labeling of endogenous, SNAP-tagged yeast proteins in whole cell extract. This method allows biochemical reactions to be followed in cell lysates in real time using colocalization single-molecule fluorescence microscopy. Labeled complexes can also be isolated from extract and characterized by SNAP tag single-molecule pull-down (SNAP-SiMPull). These approaches have proven useful for studying complex biological machines such as the spliceosome that cannot yet be reconstituted from purified components.

Evolution and characterization of a benzylguanine-binding RNA aptamer.

Repurposing the "protein-labeling toolkit" for RNA research could be a pragmatic approach for developing new RNA-labeling methods. We have evolved an RNA aptamer that tightly binds benzylguanine (bG), the key ligand for the protein SNAP-tag. The aptamer tightly binds bG fluorophores and can be purified from cellular RNA with bG agarose under native conditions.

Glutathione S-transferase M1 modulates allergen-induced NF-κB activation in asthmatic airway epithelium.

BACKGROUND: Glutathione S-transferase M1 (GSTM1) is a phase II enzyme and regulator of inflammatory signaling in airway epithelial cells. We have found upregulation of neutrophilic airway inflammation in atopic asthmatics expressing GSTM1 gene (GSTM1+) compared to GSTM1null asthmatics. We hypothesized that GSTM1 modulates NF-κB activation in bronchial epithelium in atopic asthmatics. We determined regulation of allergen-induced NF-κB activation in bronchial epithelium by GSTM1 in human atopic asthmatics in vivo. METHODS: Endobronchial biopsies and bronchoalveolar lavage fluid samples were collected from 13 GSTM1+ and 12 GSTM1null human atopic asthmatics at baseline and 24 h after segmental allergen challenge. A quantitative analysis of NF-κB activation in airway epithelium was accomplished using a polyclonal antibody against the phosphorylated p65 component of NF-κB. Elastase-positive neutrophils in the bronchial wall were quantified. RESULTS: Postallergen neutrophilia in airway subepithelium and epithelial lining fluid was greater in GSTM1+ compared to GSTM1null asthmatics. Airway eosinophilia was similar in GSTM1+ and GSTM1null asthmatics. Allergen-provoked NF-κB induction in bronchial epithelium was significantly greater in GSTM1+ compared to GSTM1null asthmatics. Activation of NF-κB activation in airway epithelial cells correlated with interleukin-8 concentrations and absolute neutrophil numbers in bronchoalveolar lavage fluid in GSTM1+ but not GSTM1null asthmatics. CONCLUSIONS: Allergen-induced neutrophilic airway inflammation in GSTM1+ asthmatics is associated with NF-κB activation in airway epithelial cells in vivo. These novel data provide a potential mechanism of the genomic link between GSTM1 polymorphism and airway neutrophilia in atopic asthma.

Glutathione S-transferase P1 Ile105Val polymorphism modulates allergen-induced airway inflammation in human atopic asthmatics in vivo.

BACKGROUND: Glutathione S-transferase P1 is a Phase II cytoprotective and detoxifying enzyme that is widely expressed in human airways. The glutathione S-transferase P1 Ile105Val polymorphism has been linked with atopic disorders and asthma. Yet, little remains known about the regulation of allergic inflammation by glutathione S-transferase P1 in human asthmatics. OBJECTIVE: To establish the effect of the glutathione S-transferase P1 Ile105Val polymorphism on allergen-induced airway inflammation and oxidant stress, and non-specific bronchial hyperresponsiveness to methacholine and reactivity to specific allergen in mild human atopic asthmatics in vivo. METHODS: Five Val(105)/Val(105) , twelve Val(105)/Ile(105) and twenty Ile(105)/Ile(105) mild atopic asthmatics underwent methacholine challenge, inhaled allergen challenge and endobronchial allergen provocation through a bronchoscope. A panel of inflammatory cytokines and chemokines, F2 -isoprostanes and isofuranes, markers of oxidative stress, thromboxane B2 and immunoglobulin E were measured in bronchoalveolar lavage fluid at baseline and 24 h after allergen instillation. RESULTS: Asthmatics with glutathione S-transferase P1 Val(105)/Val(105) compared with asthmatics with the glutathione S-transferase P1 Val(105)/Ile(105) and Ile(105)/Ile(105) had greater generation of acute phase cytokines (TNF-α, IL-6, CXCL8), IL-12, CCL11, thromboxane B2 and immunoglobulin E at 24 h after local allergen challenge. The GSTP1 genotype had no effect on airway hyperresponsiveness to methacholine and the reactivity to specific allergen. CONCLUSION: The glutathione S-transferase P1 Ile105Val polymorphism markedly modifies allergen-provoked airway inflammation in atopic asthmatics in vivo. Modulation of the biochemical milieu in response to allergen provides a mechanistic explanation for regulatory effects of glutathione S-transferase P1 polymorphism on airway pathophysiology, and may guide improvement of future therapeutic methods in human atopic asthmatics. These findings must me confirmed in a larger study population of asthmatics with various ethnicities.

Using primary care-based mental health registers to reduce social exclusion in patients with severe mental illness.

Reducing social exclusion and improving mental health are key themes within the government's modernization programme. Despite this, little is known about the social exclusion experienced by people with severe and enduring mental illness living in specific communities, apart from their over-representation amongst various socially excluded groups. This paper describes a 3-year research study that focused on the impact of introducing mental health registers into general practices in an English health district. The registers were expected to facilitate improvements in linking patients with appropriate services and so improve the healthcare they received. The overall result should have been reductions in the levels of social exclusion experienced by these patients; however, findings revealed a lack of change in unmet needs and quality of life, even amongst those in contact with a community mental health nurse.

Improving community mental health nurse targeting of people with severe and enduring mental illness: experiences from one English health district.

BACKGROUND: Successive governments have urged mental health service providers to target their attentions on people with severe and enduring mental illness (SEMI). However, community mental health teams (CMHTs) in general, and community mental health nurses (CMHNs) in particular, have been criticized for failing to meet this requirement. This paper reports selected findings from a wider study that assessed the impact of an initiative designed to facilitate service targeting: the establishment of registers of patients with SEMI in general practices throughout an English health district. The paper describes changes in the nature of community mental health nursing contacts with a sample of patients on these registers. METHODS: Six general practices were randomly selected from the 65 practices in the district and comparisons made between patients on the six mental health registers who either had, or did not have, community mental health nursing contact. These comparisons related to the year before the establishment of the registers, the year during which they were being established and the year following this. RESULTS: A total of 274 patients were included on the sample registers, with practices varying considerably in relation to proportions of mental health registered patients with community mental health nursing contact. Overall, the number of patients in contact with CMHNs was found to have decreased over time, except for those on level 2 of the Care Programme Approach. CONCLUSIONS: No evidence was found to support the hypothesis that the establishment of the registers had improved CMHN targeting of patients with SEMI. However, findings were inconclusive because shortages of CMHNs and the disruption associated with widespread service reorganization meant the registers were never fully implemented in practices during the study period.

Re-emerging syphilis in the UK: a behavioural analysis of infected individuals.

In-depth interviews with 27 individuals infected with syphilis in a recent UK outbreak (out of a total of 58 diagnosed between May 1999 and August 2000 in three city hospitals) were carried out to examine behaviour and attitudes. Most (23/27) participants were homosexual men, seven of whom were HIV positive. Between them, the 23 gay men had 1,494 different contacts in the twelve months prior to their awareness of having syphilis, but only 10% of these contacts could be named. While oral sex (usually unprotected) was the most prevalent behaviour (median = 30 partners per year), only 39% perceived unprotected oral sex as a syphilis risk (c.f. 70% for anal sex). Many gay men (61%) used gamma hydroxybutyrate (GHB) during sex as an aphrodisiac. This syphilis-infected subset of the population had high levels of unprotected and anonymous sex, which brings into question the usefulness of contact tracing to control syphilis outbreaks. The majority of partners were casual oral sex partners. More awareness is urgently needed around syphilis symptoms and risks, and risks of using drugs to reduce sexual inhibitions.

NMR structure of a DNA duplex containing nucleoside analog 1-(2'-deoxy-beta-D-ribofuranosyl)-3-nitropyrrole and the structure of the unmodified control.

The three-dimensional structures of two DNA duplexes d(CATGAGTAC). d(GTACXCATG) (1) and d(CATGAGTAC).d(GTACTCATG) (2), where X represents 1-(2'-deoxy-beta-D-ribofuranosyl)-3-nitropyrrole, were solved using high resolution nuclear magnetic resonance spectroscopy and restrained molecular dynamics. Good convergence was observed between final structures derived from A- and B-form starting geometries for both 1 and 2. Structures of 1 and 2 are right-handed duplexes within the B-form conformational regime. Furthermore, the structures of 1 and 2 are highly similar, with differences in the structures localized to the vicinity of residue 14 (X versus T). The pyrrole group of 1 is in the syn conformation and it is displaced towards the major groove. Furthermore, unlike T14 in 2, the base of X14 has reduced pi-pi stacking interactions with C13 and C15 and the nitro group of X14 is pointing out of the major groove. The structures presented here establish the basis of the thermal data of DNA duplexes containing X and should be informative during the design of improved wild card nucleobase analogs.

Selective separation of proteins from mixtures using polysaccharides.

The phase behavior of ternary (protein + dextran + solvent) and quaternary (protein 1 + protein 2 + dextran + solvent) aqueous solutions have been monitored at various ionic strengths and pHs by turbidity measurements. For the ternary systems, phase separation was favored when the solvent conditions promoted protein self-association. Bovine serum albumin (BSA) and gamma-globulin were believed to self-associate in the presence of dextran at their respective isoelectric points through the electrostatic interactions between positive and negative charged patches at their surface. The fact that phase separation for BSA became less evident on increasing the ionic strength supported this concept. In the case of lysozyme alone, self-association was promoted by the addition of electrolyte as was phase separation in the lysozyme + dextran ternary system. For this biopolymer self-association is presumably as a consequence of van der Waals and/or hydrophobic forces. The influence of dextran has been discussed in terms of polymer depletion theory where an additional attractive force is created due to the exclusion of the dextran molecules from the immediate vicinity of the protein surface. For the quaternary systems, by subtle choice of solvent conditions, BSA and globulin could be exclusively partitioned into separate liquid layers through a segregative mechanism, while BSA and lysozyme could be separated into a single highly concentrated layer through an associative mechanism.

Clinical guidelines and their implementation.

Within the UK there has been increasing interest in the development and implementation of guidelines, as the emphasis on clinical effectiveness is gathering momentum. This paper outlines some of the practical issues encountered in developing and implementing guidelines, based on experiences within Liverpool. Developing local guidelines can be a lengthy process, but that process is not a waste of time if it means there is more likely to be compliance in the end. Dissemination of guidelines alone is not enough; it needs to be combined with an appropriate implementation strategy. There is a danger of primary care being overloaded with new guidelines; there needs to be a timed strategy for their introduction. More imaginative thought needs to be put into the marketing of new ideas in order to change practice. We need to encourage the ethos amongst healthcare professionals of expecting to have to constantly update knowledge and practice.

A pilot study of a computerized assessment (AGE-PC) for the elderly in general practice.

A pilot project was undertaken in Liverpool UK, to investigate whether data useful for both general practice and public health purposes could be collected by primary care personnel using a computerized semi-structured interview (AGE-PC) for assessment of patients aged 75 and over. A high degree of variation between practices was found in the proportion of elderly assessed with AGE-PC largely related to motivation to undertake assessments and methods employed to approach patients. Considerable differences between and within practices were evident in staff attitudes to the need for detailed formal assessments and the use of computers. Several aspects of the AGE-PC package itself were identified as requiring modification including streamlining data transfer methods, adding facilities for analysis and giving users a degree of choice over content of the assessment. This technique does offer a potentially efficient means of collecting very detailed standardized data for intra- or inter-practice analysis while at the same time generating a comprehensive report for individual patients. However, in the context of the over-75 assessments it was too time-consuming. Possible future strategies are discussed such as using an initial screening stage with a self-report questionnaire to reduce numbers needing full assessment and the use of the package for the community care assessments. The pilot project highlighted some of the potential advantages and disadvantages of collecting data from primary health care for public health purposes.(ABSTRACT TRUNCATED AT 250 WORDS)

Retinal venous sheathing in optic neuritis. Its significance for the pathogenesis of multiple sclerosis.

A systematic study of the frequency of retinal vascular abnormalities and cells in the media has been made in 50 patients presenting with acute optic neuritis. Abnormalities were found in 14 (fluorescein leakage in 10, perivenous sheathing in 6, cells in the vitreous in 6 and in the anterior chamber in 4; in 2 the cells in the media were seen without vascular changes). After a mean follow up of 3.5 years multiple sclerosis (MS) had developed in 8/14 patients with vascular abnormalities and/or evidence of inflammation and in 5/32 without; the difference is significant (P less than 0.02). The occurrence of perivenular abnormalities in a region free of myelin and oligodendrocytes provides evidence that the vascular changes in MS can occur independently of contiguous demyelination, and may be the primary event in the formation of a new lesion.