Norovirus antivirals: Where are we now?

Human noroviruses inflict a significant health burden on society and are responsible for approximately 699 million infections and over 200 000 estimated deaths worldwide each year. Yet despite significant research efforts, approved vaccines or antivirals to combat this pathogen are still lacking. Safe and effective antivirals are not available, particularly for chronically infected immunocompromised individuals, and for prophylactic applications to protect high-risk and vulnerable populations in outbreak settings. Since the discovery of human norovirus in 1972, the lack of a cell culture system has hindered biological research and antiviral studies for many years. Recent breakthroughs in culturing human norovirus have been encouraging, however, further development and optimization of these novel methodologies are required to facilitate more robust replication levels, that will enable reliable serological and replication studies, as well as advances in antiviral development. In the last few years, considerable progress has been made toward the development of norovirus antivirals, inviting an updated review. This review focuses on potential therapeutics that have been reported since 2010, which were examined across at least two model systems used for studying human norovirus or its enzymes. In addition, we have placed emphasis on antiviral compounds with a defined chemical structure. We include a comprehensive outline of direct-acting antivirals and offer a discussion of host-modulating compounds, a rapidly expanding and promising area of antiviral research.     

Characteristics of Frequent Users of Emergency Medical Services in Singapore

Objectives: This study aims to describe frequent users of Emergency Medical Services (EMS) conveyed to a Singapore tertiary hospital, focusing on a comparison between younger users (age <65) and older users in diagnoses and admission rates. Methods: All patients conveyed by EMS to a tertiary hospital 4 times or more over a 1-year period in 2015 had their EMS ambulance charts and Emergency Department (ED) electronic records retrospectively analyzed (n?=?243), with admission the primary outcome. Results: The 243 frequent users were analyzed with a combined total of 1,705 visits, out of a total of 10,183 patients with 12,839 visits conveyed by EMS to Singapore General Hospital (SGH) in 2015. Younger frequent users (<65 years age) were found to be predominantly male (79.6%, p?=?0.001) and were on average responsible for more visits than elderly frequent users (8.6 vs. 5.7, p?=?0.004). Medical co-morbidities were significantly more prevalent in older users. Younger frequent users were more likely to be smokers (60.2% vs. 22.3%), heavy drinkers (51.3% vs. 8.5%), substance abusers (12.4% vs. 0.8%), and bad debtors (49.6% vs. 20.0%, p?<?0.001). A larger proportion presented with altered mental states (11.7% vs. 5.4%, p?<?0.001) and alcohol related diagnoses (34.7% vs. 5.3%, p?<?0.001). Many were picked up from public areas (45.5% vs. 19.6%, p?<?0.001), and had lower acuity triage scores at both EMS (p?<?0.001) and ED (p?=?0.001). They had lower admission rates (40.5% vs. 78.7%, p?<?0.001) and shorter length of stay (4.3 vs. 5.9 days, p?<?0.001). Univariable and multivariable analysis showed alcohol related diagnoses, history of alcohol abuse and lower triage scores were less likely to require admissions. Conclusion: Frequent EMS users consume a disproportionate amount of healthcare resources. Two broad subgroups of patients were identified: younger patients with social issues and older patients with multiple medical conditions. EMS usage by older patients was significantly associated with higher rates of admission     

Delayed enhancement magnetic resonance imaging predicts response to cardiac resynchronization therapy in patients with intraventricular dyssynchrony.

OBJECTIVES: We evaluated the ability of delayed enhancement magnetic resonance imaging (DE-MRI) to predict clinical response to cardiac resynchronization therapy (CRT). BACKGROUND: Cardiac resynchronization therapy reduces morbidity and mortality in selected heart failure patients. However, up to 30% of patients do not have a response. We hypothesized that scar burden on DE-MRI predicts response to CRT. METHODS: The DE-MRI was performed on 28 heart failure patients undergoing CRT. Patients with QRS > or =120 ms, left ventricular ejection fraction < or =35%, New York Heart Association functional class II to IV, and dyssynchrony > or =60 ms were studied. Baseline and 3-month clinical follow-up, wall motion, 6-min walk, and quality of life assessment were performed. The DE-MRI was performed 10 min after 0.20 mmol/kg intravenous gadolinium. Scar measured by planimetry was correlated with response criteria. RESULTS: Twenty-three patients completed the protocol (mean age 64.9 +/- 11.7 years), with 12 (52%) having a history of myocardial infarction. Thirteen (57%) patients met response criteria. Percent total scar was significantly higher in the nonresponse versus response group (median and interquartile range of 24.7% [18.1 to 48.7] vs. 1.0% [0.0 to 8.7], p = 0.0022) and predicted nonresponse by receiver-operating characteristic analysis (area = 0.94). At a cutoff value of 15%, percent total scar provided a sensitivity and specificity of 85% and 90%, respectively, for clinical response to CRT. Similarly, septal scar < or =40% provided a 100% sensitivity and specificity for response. Regression analysis showed linear correlations between percent total scar and change in each of the individual response criteria. CONCLUSIONS: The DE-MRI accurately predicted clinical response to CRT. This technique offers unique information in the assessment of patients referred for CRT.     

Thrombin causes subsecond changes in protein phosphorylation of platelets

We have developed a general quenched-flow approach to study platelet function as early as 0.3 seconds after stimulation. Phosphorylation of 20- and 40-kd proteins has been analyzed during the first five seconds of platelet response to thrombin from 0.1 to 5.0 U/mL and compared with the progress of aggregation and serotonin secretion. The onset time for aggregation and phosphorylation of both proteins was less than one second, although with lowest (less than 0.5 U/mL) thrombin levels, a lag of up to 0.6 seconds occurred before 40K phosphorylation increased. The thrombin sensitivity of aggregation and 20K phosphorylation was approximately twice that of 40K phosphorylation, with Ka values of 0.51 and 0.53 v 1.10 U/mL, respectively. External calcium was necessary for maximal 20K phosphorylation, since EDTA inhibited this by 30%. The 40K phosphorylation was not affected by EDTA. Platelet activation by thrombin thus induced biochemical changes well before one second. The quenched-flow approach may help to reveal relationships between phospholipase activation, calcium fluxes, and protein phosphorylation during these early periods of platelet function.