Host factors associated with the Sindbis virus RNA-dependent RNA polymerase: role for G3BP1 and G3BP2 in virus replication.

Sindbis virus (SINV) is the prototype member of the Alphavirus genus, whose members cause severe human diseases for which there is no specific treatment. To ascertain host factors important in the replication of the SINV RNA genome, we generated a SINV expressing nsP4, the viral RNA-dependent RNA polymerase, with an in-frame 3xFlag epitope tag. Proteomic analysis of nsP4-containing complexes isolated from cells infected with the tagged virus revealed 29 associated host proteins. Of these, 10 proteins were associated only at a later time of infection (12 h), 14 were associated both early and late, and five were isolated only at the earlier time (6 h postinfection). These results demonstrate the dynamic nature of the virus-host interaction that occurs over the course of infection and suggest that different host proteins may be required for the multiple functions carried out by nsP4. Two related proteins found in association with nsP4 at both times of infection, GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) and G3BP2 were also previously identified as associated with SINV nsP2 and nsP3. We demonstrate a likely overlapping role for these host factors in limiting SINV replication events. The present study also identifies 10 host factors associated with nsP4 6 h after infection that were not found to be associated with nsP2 or nsP3. These factors are candidates for playing important roles in the RNA replication process. Identifying host factors essential for replication should lead to new strategies to interrupt alphavirus replication.

What is the cost of delayed diagnosis of bile acid malabsorption and bile acid diarrhoea?

INTRODUCTION: 75Selenium taurocholic acid (SeHCAT) scanning diagnoses bile acid malabsorption/bile acid diarrhoea (BAM/BAD) and defines optimal treatment. Approximately 2% of the population have BAM/BAD. AIM: To evaluate the cost of delayed diagnosis of BAM/BAD. METHODS: Patients' notes who underwent SeHCAT scanning in three hospitals over a 1-year period were reviewed retrospectively. Scan results and treatment response were recorded. Package-of-care costs were calculated using costing tools from the National Institute for Health and Care Excellence and from United Lincolnshire Hospitals Trust business unit. RESULTS: Between June 2016 and May 2017, 19 men and 37 women (median age 58 (range 19-83)) of 3860 new patients seen in gastroenterology clinics were referred for SeHCAT scanning. Sixty-four per cent of scans were abnormal: 13 demonstrated severe (<5% 7-day SeHCAT retention), 13 moderate (5%-10%), 5 mild (10%-15%) and 5 borderline (15%-20%) BAD/BAM. Likely causes included primary BAD (n=16), cholecystectomy (n=13), inflammatory bowel disease (n=4) and other (n=3). If SeHCAT scanning was ordered at first consultation (n=11), patients reported 24 months (median) of symptoms (range 6-360) and the median diagnostic package-of-care cost was £811.40 (95% CI £625.59 to £1508.20). If SeHCAT scanning was booked later (n=25), patients reported symptoms for 30 months (median, range 0.5-360) and the cost was £1568.31 (95% CI £1200.55 to £1713.18). Following diagnosis, treatment led to symptom improvement (n=24), no change/deterioration (n=3) and not reported (n=9). CONCLUSIONS: SeHCAT is underused. Late diagnosis leads to unnecessary demands for other services and treatment delay. Early diagnosis achieves health benefits while reducing costs.

Teaching students behavior change skills: description and assessment of a new Motivational interviewing curriculum.

BACKGROUND: One of the US government health goals outlined in the Healthy People 2010 document (2000) is to increase the number of physicians who counsel their patients regarding unhealthy behaviors. Studies have shown a low rate of physicians provide smoking cessation counseling. We introduced a motivational interviewing curriculum into our medical school for first and third year students and then evaluated the effect of this curriculum on third year students counseling skills. METHODS: The motivational interviewing curriculum was comprised of a lecture series and small group teaching with practice in role plays. The effectiveness of the curriculum was evaluated by student performance in a videotaped interview with a standardized patient who portrayed a smoker. The interview was rated using the Motivational Interviewing Treatment Integrity scoring tool (MITI). The MITI assesses 6 criteria: empathy, MI spirit (autonomy, evocation and collaboration), MI adherence (asking permission, affirmation, emphasis of control and support), MI non adherence (advise, confront and direct), the types of questions (open or closed) and the number of reflections. Secondary outcomes included a knowledge exam related to motivational interviewing and students' evaluations of the effectiveness of the motivational interviewing curriculum. RESULTS: Analysis of the MITI scores showed that students reached a proficiency level on the rate of reflections, were just below proficiency in assessment of empathy and motivational interviewing spirit and substantially below proficiency in the percent of open ended questions. These proficiency scores were for professional counselors but nevertheless provided us with information on the effectiveness of the new curriculum and where the focus of our teaching should be. On the optional evaluation of the first-year MI curriculum by 112 students, 83% felt that the MI curriculum had helped them be more comfortable in discussing behavior change with patients and 98% felt it was an important skill for physicians to have.