A fluorescent reporter system enables spatiotemporal analysis of host cell modification during herpes simplex virus-1 replication.

Herpesviruses are large and complex viruses that have a long history of coevolution with their host species. One important factor in the virus-host interaction is the alteration of intracellular morphology during viral replication with critical implications for viral assembly. However, the details of this remodeling event are not well understood, in part because insufficient tools are available to deconstruct this highly heterogeneous process. To provide an accurate and reliable method of investigating the spatiotemporal dynamics of virus-induced changes to cellular architecture, we constructed a dual-fluorescent reporter virus that enabled us to classify four distinct stages in the infection cycle of herpes simplex virus-1 at the single cell level. This timestamping method can accurately track the infection cycle across a wide range of multiplicities of infection. We used high-resolution fluorescence microscopy analysis of cellular structures in live and fixed cells in concert with our reporter virus to generate a detailed and chronological overview of the spatial and temporal reorganization during viral replication. The highly orchestrated and striking relocation of many organelles around the compartments of secondary envelopment during transition from early to late gene expression suggests that the reshaping of these compartments is essential for virus assembly. We furthermore find that accumulation of HSV-1 capsids in the cytoplasm is accompanied by fragmentation of the Golgi apparatus with potential impact on the late steps of viral assembly. We anticipate that in the future similar tools can be systematically applied for the systems-level analysis of intracellular morphology during replication of other viruses.

Safety of insertion of the copper IUD and LNG-IUS in nulliparous women: a systematic review.

OBJECTIVE: Intrauterine contraception (IUC) is one of the more effective contraceptive methods for women at highest risk of unintended pregnancy. This includes younger, often nulliparous, women; however, uptake has been relatively low in this group. METHODS: In February 2017 we conducted a systematic review of randomised controlled trials, prospective and retrospective observational studies to identify barriers to IUC use in nulliparous women. RESULTS: Study quality was poor. No differences in rates of infection or expulsions between nulliparous and parous were seen. Fertility rates following removal appeared no different from the general population. Higher rates insertion difficulty, insertion failure and pain during insertion were observed in nulliparous women. CONCLUSION: A long-acting reversible contraceptive method such as IUC reduces the risk of unintended pregnancy since user failure is minimised. Evidence-based information about the advantages and disadvantages of IUC is required to inform decision-making and dispel any myths and misperceptions. Potential barriers to IUC use in nulliparous women, particularly concerns around infection, significantly higher rates of device expulsion and adverse effects on fertility, do not appear to be justified. IUC is appropriate for all medically-eligible women, including nulliparous women, and should be included in the range of contraceptive options discussed during counselling.

The misuse and overuse of non-sterile gloves: application of an audit tool to define the problem.

BACKGROUND: The use of non-sterile gloves (NSG) has become routine in the delivery of health care, often for procedures for which they are not required; their use may increase the risk of cross contamination and is generally not integrated into hand hygiene audit. This paper describes a small-scale application and validation of an observational audit tool devised to identify inappropriate use of NSG and potential for cross contamination. METHODS: Two observers simultaneously observed the use of NSG during episodes of care in an acute hospital setting. The inter-rater reliability (IRR) of the audit tool was measured corrected for chance agreement using Kappa. RESULTS: A total of 22 episodes of care using NSG were observed. In 68.6% (24/35) of procedures there was no contact with blood/body fluid; in 54.3% (19/35) NSG-use was inappropriate. The IRR was 100% for eight of 12 components of the tool. For hand hygiene before and after NSG removal it was 82% (Kappa = 0.72) and 95% (Kappa = 0.87). CONCLUSIONS: In this small-scale application of a glove-use audit tool we demonstrated over-use and misuse of NSG and potential for cross transmission on gloved hands. The audit tool provides an effective mechanism for integrating glove use into the audit of hand hygiene behaviour.

Simulation can contribute a part of cardiorespiratory physiotherapy clinical education: two randomized trials.

INTRODUCTION: Simulated learning environments (SLEs) are used worldwide in health professional education, including physiotherapy, to train certain attributes and skills. To date, no randomized controlled trial (RCT) has evaluated whether education in SLEs can partly replace time in the clinical environment for physiotherapy cardiorespiratory practice. METHODS: Two independent single-blind multi-institutional RCTs were conducted in parallel using a noninferiority design. Participants were volunteer physiotherapy students (RCT 1, n = 176; RCT 2, n = 173) entering acute care cardiorespiratory physiotherapy clinical placements. Two SLE models were investigated as follows: RCT 1, 1 week in SLE before 3 weeks of clinical immersion; RCT 2, 2 weeks of interspersed SLE/clinical immersion (equivalent to 1 SLE week) within the 4-week clinical placement. Students in each RCT were stratified on academic grade and randomly allocated to an SLE plus clinical immersion or clinical immersion control group. The primary outcome was competency to practice measured in 2 clinical examinations using the Assessment of Physiotherapy Practice. Secondary outcomes were student perception of experience and clinical educator and patient rating of student performance. RESULTS: There were no significant differences in student competency between the SLE and control groups in either RCT, although students in the interspersed group (RCT 2) achieved a higher score in 5 of 7 Assessment of Physiotherapy Practice standards (all P < 0.05). Students rated the SLE experience positively. Clinical educators and patients reported comparability between groups. CONCLUSIONS: An SLE can replace clinical time in cardiorespiratory physiotherapy practice. Part education in the SLE satisfied clinical competency requirements, and all stakeholders were satisfied.

Can simulation replace part of clinical time? Two parallel randomised controlled trials.

CONTEXT: Education in simulated learning environments (SLEs) has grown rapidly across health care professions, yet no substantive randomised controlled trial (RCT) has investigated whether SLEs can, in part, substitute for traditional clinical education. METHODS: Participants were physiotherapy students (RCT 1, n = 192; RCT 2, n = 178) from six Australian universities undertaking clinical education in an ambulatory care setting with patients with musculoskeletal disorders. A simulated learning programme was developed as a replica for clinical education in musculoskeletal practice to replace 1 week of a 4-week clinical education placement. Two SLE models were designed. Model 1 provided 1 week in the SLE, followed by 3 weeks in clinical immersion; Model 2 offered training in the SLE in parallel with clinical immersion during the first 2 weeks of the 4-week placement. Two single-blind, multicentre RCTs (RCT 1, Model 1; RCT 2, Model 2) were conducted using a non-inferiority design to determine if the clinical competencies of students part-educated in SLEs would be any worse than those of students educated fully in traditional clinical immersion. The RCTs were conducted simultaneously, but independently. Within each RCT, students were stratified on academic score and randomised to either the SLE group or the control ('Traditional') group, which undertook 4 weeks of traditional clinical immersion. The primary outcome measure was a blinded assessment of student competency conducted over two clinical examinations at week 4 using the Assessment of Physiotherapy Practice (APP) tool. RESULTS: Students' achievement of clinical competencies was no worse in the SLE groups than in the Traditional groups in either RCT (Margin [Δ] ≥ 0.4 difference on APP score; RCT 1: 95% CI - 0.07 to 0.17; RCT 2: 95% CI - 0.11 to 0.16). CONCLUSIONS: These RCTs provide evidence that clinical education in an SLE can in part (25%) replace clinical time with real patients without compromising students' attainment of the professional competencies required to practise.

Phase 1 study of HPV16-specific immunotherapy with E6E7 fusion protein and ISCOMATRIX adjuvant in women with cervical intraepithelial neoplasia.

PURPOSE: Persistent infection of cervical epithelium with "high risk" human papillomavirus (HPV) results in cervical intraepithelial neoplasia (CIN) from which squamous cancer of the cervix can arise. A study was undertaken to evaluate the safety and immunogenicity of an HPV16 immunotherapeutic consisting of a mixture of HPV16 E6E7 fusion protein and ISCOMATRIX adjuvant (HPV16 Immunotherapeutic) for patients with CIN. EXPERIMENTAL DESIGN: Patients with CIN (n = 31) were recruited to a randomised blinded placebo controlled dose ranging study of immunotherapy. RESULTS: Immunotherapy was well tolerated. Immunised subjects developed HPV16 E6E7 specific immunity. Antibody, delayed type hypersensitivity, in vitro cytokine release, and CD8 T cell responses to E6 and E7 proteins were each significantly greater in the immunised subjects than in placebo recipients. Loss of HPV16 DNA from the cervix was observed in some vaccine and placebo recipients. CONCLUSIONS: The HPV16 Immunotherapeutic comprising HPV16E6E7 fusion protein and ISCOMATRIX adjuvant is safe and induces vaccine antigen specific cell mediated immunity.

Women teaching women's health: issues in the establishment of a clinical teaching associate program for the well woman check.

The impact of screening programs for cervical cancer would be increased with the greater participation of currently underscreened women. Training for medical students and doctors in the fine technical and communication skills required in breast and gynaecological examinations would improve participation by increasing the confidence and skill of doctors in raising the issue of screening, thereby making the examination a more positive experience for women. Gynaecology Teaching Associate (GTA) programs, using specially trained standardized patients, have been used in over 90% of American and Canadian medical schools for more than ten years to provide such training. Australia has been slow to adopt this teaching method. A Clinical Teaching Associates in Gynaecology program (CTA) was first established in 1996 by the Department of Obstetrics and Gynaecology at the University of Queensland, building on the Pap test program from Adelaide. Other medical schools subsequently introduced such programs and in 2000, the Department of General Practice, University of Melbourne, established a CTA program based on the Queensland program, with a grant from PapScreen Victoria. This paper describes the methods of recruitment and training of CTAs, use of CTAs in the medical course, preliminary evaluation, and ethical and other issues in the Melbourne and Queensland University programs.

Dissemination of an efficacious antenatal smoking cessation program in public hospitals in Australia: a randomized controlled trial.

This study investigated the impact of a behaviorally based intervention designed to increase the number of hospitals that routinely provide effective smoking cessation programs for pregnant women. In Queensland, Australia, 70 publicly funded hospitals were matched on numbers of births and maternal socioeconomic status and randomly allocated to an awareness-only intervention group or a behaviorally based intervention group. Success was defined as the routine offer of an evidence-based smoking cessation program to at least 80% of the pregnant clients who smoke. At 1 month, 65% of the behaviorally based intervention hospitals agreed to provide materials about smoking cessation programs for their antenatal patients, compared with 3% of the awareness-only hospitals. After 1 year, 43% of the intervention hospitals still provided the material, compared with 9% of the awareness-only hospitals. These findings show that a brief intervention to hospitals can encourage antenatal staff to provide smoking cessation materials to pregnant women.