Establishing severe acute respiratory infection (SARI) surveillance in a sentinel hospital, Ireland, 2021 to 2022.

BackgroundIn 2020, due to the COVID-19 pandemic, the European Centre for Disease Prevention and Control (ECDC) accelerated development of European-level severe acute respiratory infection (SARI) surveillance.AimWe aimed to establish SARI surveillance in one Irish hospital as part of a European network E-SARI-NET.MethodsWe used routine emergency department records to identify cases in one adult acute hospital. The SARI case definition was adapted from the ECDC clinical criteria for a possible COVID-19 case. Clinical data were collected using an online questionnaire. Cases were tested for SARS-CoV-2, influenza and respiratory syncytial virus (RSV), including whole genome sequencing (WGS) on SARS-CoV-2 RNA-positive samples and viral characterisation/sequencing on influenza RNA-positive samples. Descriptive analysis was conducted for SARI cases hospitalised between July 2021 and April 2022.ResultsOverall, we identified 437 SARI cases, the incidence ranged from two to 28 cases per week (0.7-9.2/100,000 hospital catchment population). Of 431 cases tested for SARS-CoV-2 RNA, 226 (52%) were positive. Of 349 (80%) cases tested for influenza and RSV RNA, 15 (4.3%) were positive for influenza and eight (2.3%) for RSV. Using WGS, we identified Delta- and Omicron-dominant periods. The resource-intensive nature of manual clinical data collection, specimen management and laboratory supply shortages for influenza and RSV testing were challenging.ConclusionWe successfully established SARI surveillance as part of E-SARI-NET. Expansion to additional sentinel sites is planned following formal evaluation of the existing system. SARI surveillance requires multidisciplinary collaboration, automated data collection where possible, and dedicated personnel resources, including for specimen management.

Comparison of the performance of an amplicon sequencing assay based on Oxford Nanopore technology to real-time PCR assays for detecting bacterial biodefense pathogens.

BACKGROUND: The state-of-the-art in nucleic acid based biodetection continues to be polymerase chain reaction (PCR), and many real-time PCR assays targeting biodefense pathogens for biosurveillance are in widespread use. These assays are predominantly singleplex; i.e. one assay tests for the presence of one target, found in a single organism, one sample at a time. Due to the intrinsic limitations of such tests, there exists a critical need for high-throughput multiplex assays to reduce the time and cost incurred when screening multiple targets, in multiple pathogens, and in multiple samples. Such assays allow users to make an actionable call while maximizing the utility of the small volumes of test samples. Unfortunately, current multiplex real-time PCR assays are limited in the number of targets that can be probed simultaneously due to the availability of fluorescence channels in real-time PCR instruments. RESULTS: To address this gap, we developed a pipeline in which the amplicons produced by a 14-plex end-point PCR assay using spiked samples were subsequently sequenced using Nanopore technology. We used bar codes to sequence multiple samples simultaneously, leading to the generation and subsequent analysis of sequence data resulting from a short sequencing run time (< 10 min). We compared the limits of detection (LoD) of real-time PCR assays to Oxford Nanopore Technologies (ONT)-based amplicon sequencing and estimated the sample-to-answer time needed for this approach. Overall, LoDs determined from the first 10 min of sequencing data were at least one to two orders of magnitude lower than real-time PCR. Given enough time, the amplicon sequencing approach is approximately 100 times more sensitive than real-time PCR, with detection of amplicon specific reads even at the lowest tested spiking concentration (around 2.5-50 Colony Forming Units (CFU)/ml). CONCLUSIONS: Based on these results, we propose amplicon sequencing assay as a viable alternative to replace the current real-time PCR based singleplex assays for higher throughput biodefense applications. We note, however, that targeted amplicon specific reads were not detectable even at the highest tested spike concentrations (2.5 X 104-5.0 X105 CFU/ml) without an initial amplification step, indicating that PCR is still necessary when utilizing this protocol.

Early detection of Ebola virus proteins in peripheral blood mononuclear cells from infected mice.

BACKGROUND: Detection of viral ribo-nucleic acid (RNA) via real-time polymerase chain reaction (RT-PCR) is the gold standard for the detection of Ebola virus (EBOV) during acute infection. However, the earliest window for viral RNA detection in blood samples is 48-72 h post-onset of symptoms. Therefore, efforts to develop additional orthogonal assays using complementary immunological and serological technologies are still needed to provide simplified methodology for field diagnostics. Furthermore, unlike RT-PCR tests, immunoassays that target viral proteins and/or early host responses are less susceptible to sequence erosion due to viral genetic drift. Although virus is shed into the bloodstream from infected cells, the wide dynamic range of proteins in blood plasma makes this a difficult sample matrix for the detection of low-abundant viral proteins. We hypothesized that the isolation of peripheral blood mononuclear cells (PBMCs), which are the first cellular targets of the Ebola virus (EBOV), may provide an enriched source of viral proteins. METHODS: A mouse infection model that employs a mouse-adapted EBOV (MaEBOV) was chosen as a proof-of-principal experimental paradigm to determine if viral proteins present in PBMCs can help diagnose EBOV infection pre-symptomatically. We employed a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) platform to provide both high sensitivity and specificity for the detection and relative quantitation of viral proteins in PBMCs collected during MaEBOV infection. Blood samples pooled from animals at the post-infection time-points were used to determine the viral load by RT-PCR and purify PBMCs. RESULTS: Using quantitative LC-MS/MS, we detected two EBOV proteins (vp40 and nucleoprotein) in samples collected on Day 2 post-infection, which was also the first day of detectable viremia via RT-PCR. These results were confirmed via western blot which was performed on identical PBMC lysates from each post-infection time point. CONCLUSIONS: While mass spectrometry is not currently amenable to field diagnostics, these results suggest that viral protein enrichment in PBMCs in tandem with highly sensitive immunoassays platforms, could lead to the development of a rapid, high-throughput diagnostic platform for pre-symptomatic detection of EBOV infection.

A comprehensive multi-omics approach uncovers adaptations for growth and survival of Pseudomonas aeruginosa on n-alkanes.

BACKGROUND: Examination of complex biological systems has long been achieved through methodical investigation of the system's individual components. While informative, this strategy often leads to inappropriate conclusions about the system as a whole. With the advent of high-throughput "omic" technologies, however, researchers can now simultaneously analyze an entire system at the level of molecule (DNA, RNA, protein, metabolite) and process (transcription, translation, enzyme catalysis). This strategy reduces the likelihood of improper conclusions, provides a framework for elucidation of genotype-phenotype relationships, and brings finer resolution to comparative genomic experiments. Here, we apply a multi-omic approach to analyze the gene expression profiles of two closely related Pseudomonas aeruginosa strains grown in n-alkanes or glycerol. RESULTS: The environmental P. aeruginosa isolate ATCC 33988 consumed medium-length (C10-C16) n-alkanes more rapidly than the laboratory strain PAO1, despite high genome sequence identity (average nucleotide identity >99%). Our data shows that ATCC 33988 induces a characteristic set of genes at the transcriptional, translational and post-translational levels during growth on alkanes, many of which differ from those expressed by PAO1. Of particular interest was the lack of expression from the rhl operon of the quorum sensing (QS) system, resulting in no measurable rhamnolipid production by ATCC 33988. Further examination showed that ATCC 33988 lacked the entire lasI/lasR arm of the QS response. Instead of promoting expression of QS genes, ATCC 33988 up-regulates a small subset of its genome, including operons responsible for specific alkaline proteases and sphingosine metabolism. CONCLUSION: This work represents the first time results from RNA-seq, microarray, ribosome footprinting, proteomics, and small molecule LC-MS experiments have been integrated to compare gene expression in bacteria. Together, these data provide insights as to why strain ATCC 33988 is better adapted for growth and survival on n-alkanes.

Argininosuccinate synthetase 1 depletion produces a metabolic state conducive to herpes simplex virus 1 infection.

Herpes simplex virus 1 (HSV-1) infection triggers specific metabolic changes in its host cell. To explore the interactions between cellular metabolism and HSV-1 infection, we performed an siRNA screen of cellular metabolic genes, measuring their effect on viral replication. The screen identified multiple enzymes predicted to influence HSV-1 replication, including argininosuccinate synthetase 1 (AS1), which consumes aspartate as part of de novo arginine synthesis. Knockdown of AS1 robustly enhanced viral genome replication and the production of infectious virus. Using high-resolution liquid chromatography-mass spectrometry, we found that the metabolic phenotype induced by knockdown of AS1 in human fibroblasts mimicked multiple aspects of the metabolic program observed during HSV-1 infection, including an increase in multiple nucleotides and their precursors. Together with the observation that AS1 protein and mRNA levels decrease during wild-type infection, this work suggests that reduced AS1 activity is partially responsible for the metabolic program induced by infection.

Herpes simplex virus 1 infection activates poly(ADP-ribose) polymerase and triggers the degradation of poly(ADP-ribose) glycohydrolase.

Herpes simplex virus 1 infection triggers multiple changes in the metabolism of host cells, including a dramatic decrease in the levels of NAD(+). In addition to its role as a cofactor in reduction-oxidation reactions, NAD(+) is required for certain posttranslational modifications. Members of the poly(ADP-ribose) polymerase (PARP) family of enzymes are major consumers of NAD(+), which they utilize to form poly(ADP-ribose) (PAR) chains on protein substrates in response to DNA damage. PAR chains can subsequently be removed by the enzyme poly(ADP-ribose) glycohydrolase (PARG). We report here that the HSV-1 infection-induced drop in NAD(+) levels required viral DNA replication, was associated with an increase in protein poly(ADP-ribosyl)ation (PARylation), and was blocked by pharmacological inhibition of PARP-1/PARP-2 (PARP-1/2). Neither virus yield nor the cellular metabolic reprogramming observed during HSV-1 infection was altered by the rescue or further depletion of NAD(+) levels. Expression of the viral protein ICP0, which possesses E3 ubiquitin ligase activity, was both necessary and sufficient for the degradation of the 111-kDa PARG isoform. This work demonstrates that HSV-1 infection results in changes to NAD(+) metabolism by PARP-1/2 and PARG, and as PAR chain accumulation can induce caspase-independent apoptosis, we speculate that the decrease in PARG levels enhances the auto-PARylation-mediated inhibition of PARP, thereby avoiding premature death of the infected cell.

Divergent effects of human cytomegalovirus and herpes simplex virus-1 on cellular metabolism.

Viruses rely on the metabolic network of the host cell to provide energy and macromolecular precursors to fuel viral replication. Here we used mass spectrometry to examine the impact of two related herpesviruses, human cytomegalovirus (HCMV) and herpes simplex virus type-1 (HSV-1), on the metabolism of fibroblast and epithelial host cells. Each virus triggered strong metabolic changes that were conserved across different host cell types. The metabolic effects of the two viruses were, however, largely distinct. HCMV but not HSV-1 increased glycolytic flux. HCMV profoundly increased TCA compound levels and flow of two carbon units required for TCA cycle turning and fatty acid synthesis. HSV-1 increased anapleurotic influx to the TCA cycle through pyruvate carboxylase, feeding pyrimidine biosynthesis. Thus, these two related herpesviruses drive diverse host cells to execute distinct, virus-specific metabolic programs. Current drugs target nucleotide metabolism for treatment of both viruses. Although our results confirm that this is a robust target for HSV-1, therapeutic interventions at other points in metabolism might prove more effective for treatment of HCMV.

Effects of depressive and other psychiatric disorders on anticoagulation control in a pharmacist-managed anticoagulation clinic.

BACKGROUND: The impact of psychiatric disorders on International Normalized Ratio (INR) control and adverse events for patients receiving warfarin has not been fully elucidated. OBJECTIVE: To determine the effect of depressive and other psychiatric disorders on anticoagulation control in a pharmacist-managed anticoagulation clinic. METHODS: A retrospective chart review evaluated outcomes of patients with no history of psychiatric disorders and compare it with that of patients with either a history of depression or any form of psychiatric disorder. Data was obtained from patient medical records over a 24-month period. The primary outcome was a comparison of time in therapeutic range, calculated using 3 separate methods (percentage of INRs in therapeutic range, a modified Rosendaal's linear interpolation, and mean INR in goal). RESULTS: A total of 151 patients met the inclusion criteria (control = 79, psychiatric disorders = 72 patients). Control patients had a significantly greater proportion of INRs in the goal range compared with either the depression or psychiatric disorders groups (control, 55.7%; depression, 43.5%; psychiatric, 45.8%). Utilizing the Rosendaal's method, patients with psychiatric disorders were in the goal range significantly less often than those in the control group (53.0% vs 61.3%). No differences were seen when adjusting for multiple comparisons or when comparing the control and depression groups (54.5% vs 61.3%). There was no difference between the groups when comparing percentages of patients with a mean INR in their goal range. CONCLUSIONS: Patients with psychiatric disorders who take warfarin may spend less time in the therapeutic range.

Draft Genome Assemblies of Phage AP50c-Resistant Derivatives of Bacillus anthracis Sterne Strain 7702 Lacking Plasmid pXO2.

Mutants of the attenuated Bacillus anthracis (Sterne) strain 7702 that are resistant to phage AP50c have been previously described. Here, we report the draft genome assemblies of the parent strain, several phage-resistant derivatives, and mutants of genes in the pathways for synthesis and assembly of the S-layer.

Genetic evidence for the interaction between Bacillus anthracis-encoded phage receptors and their cognate phage-encoded receptor binding proteins.

Bacteriophages such as γ and AP50c have been shown to infect strains of Bacillus anthracis with high specificity, and this feature has been exploited in the development of bacterial detection assays. To better understand the emergence of phage resistance, and thus the potential failure of such assays, it is important to identify the host and phage receptors necessary for attachment and entry. Using genetic approaches, the bacterial receptors of AP50c and γ have been identified as sap and GamR, respectively. A second AP50c-like phage, Wip1, also appears to use sap as a receptor. In parallel with this work, the cognate phage-encoded receptor binding proteins (RBPs) have also been identified (Gp14 for γ, P28 for AP50c, and P23 for Wip1); however, the strength of evidence supporting these protein-protein interactions varies, necessitating additional investigation. Here, we present genetic evidence further supporting the interaction between sap and the RBPs of AP50c and Wip1 using fluorescently tagged proteins and a panel of B. anthracis mutants. These results showed that the deletion of the sap gene, as well as the deletion of csaB, whose encoded protein anchors sap to the bacterial S-layer, resulted in the loss of RBP binding. Binding could then be rescued by expressing these genes in trans. We also found that the RBP of the γ-like prophage λBa03 relied on csaB activity for binding, possibly by a different mechanism. RBPλBa03 binding to B. anthracis cells was also unique in that it was not ablated by heat inactivation of vegetative cells, suggesting that its receptor is still functional following incubation at 98°C. These results extend our understanding of the diverse attachment and entry strategies used by B. anthracis phages, enabling future assay development.

Routine Decontamination of Surfaces Relevant to Working Dogs: Neutralization of Superficial Coronavirus Contamination.

Given the increased deployment of working dogs to settings with pathogenic biological agents, a safe, effective, and logistically feasible surface decontamination protocol is essential to protect both the animals and their human handlers. Our group previously found that superficial contamination on surfaces relevant to the working dog community, including leashes and toys, could be significantly reduced using a standardized wiping protocol with various cleansing products. To expand upon this work, we analyzed the ability of this protocol to decontaminate surface-deposited bovine coronavirus, which was used as a BSL2 surrogate for SARS-CoV-2. Unsurprisingly, the physical characteristics of a given surface, including porosity and texture, had a significant effect on the ability to recover viable virus remaining on the surface post treatment. After correcting for these differences, however, wiping with 70% isopropyl alcohol (IPA) and 0.5% chlorhexidine performed best, reducing viral titers by >3 log on plastic bumper toys and nylon collars, and by >2 log on rubber toys and tennis balls. Leather leashes and Velcro proved more difficult to decontaminate, but both still showed significant loss of viral contamination following wiping with IPA or chlorhexidine. This work (i) validates the utility of a simple protocol for the neutralization of viruses on several surfaces, (ii) identifies materials that are more difficult to decontaminate, which should, thus, be considered for removal from field use, and (iii) highlights the need for further development of protocols testing porous or textured surfaces.

Optimization of Oxford Nanopore Technology Sequencing Workflow for Detection of Amplicons in Real Time Using ONT-DART Tool.

An optimized, well-tested and validated targeted genomic sequencing-based high-throughput assay is currently not available ready for routine biodefense and biosurveillance applications. Earlier, we addressed this gap by developing and establishing baseline comparisons of a multiplex end-point Polymerase Chain Reaction (PCR) assay followed by Oxford Nanopore Technology (ONT) based amplicon sequencing to real time PCR and customized data processing. Here, we expand upon this effort by identifying the optimal ONT library preparation method for integration into a novel software platform ONT-DART (ONT-Detection of Amplicons in Real-Time). ONT-DART is a dockerized, real-time, amplicon-sequence analysis workflow that is used to reproducibly process and filter read data to support actionable amplicon detection calls based on alignment metrics, within sample statistics, and no-template control data. This analysis pipeline was used to compare four ONT library preparation protocols using R9 and Flongle (FL) flow cells. The two 4-Primer methods tested required the shortest preparation times (5.5 and 6.5 h) for 48 libraries but provided lower fidelity data. The Native Barcoding and Ligation methods required longer preparation times of 8 and 12 h, respectively, and resulted in higher overall data quality. On average, data derived from R9 flow cells produced true positive calls for target organisms more than twice as fast as the lower throughput FL flow cells. These results suggest that utilizing the R9 flowcell with an ONT Native Barcoding amplicon library method in combination with ONT-DART platform analytics provides the best sequencing-based alternative to current PCR-based biodetection methods.

The META tool optimizes metagenomic analyses across sequencing platforms and classifiers.

A major challenge in the field of metagenomics is the selection of the correct combination of sequencing platform and downstream metagenomic analysis algorithm, or "classifier". Here, we present the Metagenomic Evaluation Tool Analyzer (META), which produces simulated data and facilitates platform and algorithm selection for any given metagenomic use case. META-generated in silico read data are modular, scalable, and reflect user-defined community profiles, while the downstream analysis is done using a variety of metagenomic classifiers. Reported results include information on resource utilization, time-to-answer, and performance. Real-world data can also be analyzed using selected classifiers and results benchmarked against simulations. To test the utility of the META software, simulated data was compared to real-world viral and bacterial metagenomic samples run on four different sequencers and analyzed using 12 metagenomic classifiers. Lastly, we introduce "META Score": a unified, quantitative value which rates an analytic classifier's ability to both identify and count taxa in a representative sample.

Decreasing the stigma surrounding alcohol use disorder.

INTRODUCTION: There is a paucity of data on educational interventions that prepare students to mitigate the stigma or burden of alcohol use disorder. The objectives of this study were to (1) assess the impact of an interprofessional symposium on personal knowledge and stigma of alcohol use disorder and (2) inform future educational models. METHODS: The symposium highlighted the impact of alcohol at one private Midwestern university and reviewed the pharmacology of alcohol, diagnostic criteria for alcohol use disorder, and treatment for alcohol use disorder. Prior to and after the symposium, participants were given nine statements (two knowledge-based and seven stigma-based) about alcohol use disorder. Agreement with each statement was measuring on a five-point rating scale, and responses were collapsed into three categories: 1 = low stigma/high understanding, 2 = neutral, and 3 = high stigma/low understanding. Change between response categories before and after the symposium were analyzed using a Wilcoxon signed-ranked test (W). RESULTS: A total of 87 responses were collected pre-symposium and 45 responses were collected post-symposium. Both knowledge-based statements showed an increase in individual respondent understanding of alcohol use disorder as a disease. All stigma-based statements conveyed a decrease in individual respondent stigma of alcohol use disorder as a disease. Test statistics (Z) for significant items raged between Z = 3 to 5, P < .05. CONCLUSIONS: The symposium was successful at conveying positive changes in attitudes toward alcohol use disorder.

Incidence of a negative hidden curriculum, cynicism, and burnout within pharmacy resident education: A nationwide survey.

INTRODUCTION: The term "hidden curriculum" (HC) is a set of ethical, moral, and value-based teachings communicated in a non-explicit manner. Recent literature has described increasing awareness of the prevalence of the HC and potential negative impact on medical learners; however, this information is lacking in pharmacy resident education. Consequently, we conducted a survey study of United States pharmacy residents to learn their perceptions concerning the HC in pharmacy residency training. METHODS: A nationwide survey of pharmacy residents was conducted in June 2019. The survey assessed the following: presence of negative HC (score 0 to 80), cynicism (score 0 to 25), burnout via Maslach Burnout Inventory depersonalization (MBI-D) (range 0 to 30), and emotional exhaustion via Maslach Burnout Inventory emotional exhaustion (MBI-EE) (range 0 to 54). Higher scores represent increased occurrences of each domain. RESULTS: The mean HC score was 20 (SD 14.7), mean cynicism score was 9 (SD 5.5), MBI-D was 5.5 (SD 4.5), and MBI-EE was 24.2 (SD 12.4). Of those completing an MBI score, 40.4% (82/203) reported burnout in one area, while 15.8% (32/203) reported burnout in both areas. Residents reporting burnout had higher mean HC and cynicism scores. CONCLUSIONS: Awareness to develop and grow cultures that minimize the presence of a negative HC is essential to improve postgraduate pharmacy training.

Routine Decontamination of Working Canines: A Study on the Removal of Superficial Gross Contamination.

Odor detection canines are a valuable resource used by multiple agencies for the sensitive detection of explosives, narcotics, firearms, agricultural products, and even human bodies. These canines and their handlers are frequently deployed to pathogen-contaminated environments or to work in close proximity with potentially sick individuals. Appropriate decontamination protocols must be established to mitigate both canine and handler exposure in these scenarios. Despite this potential risk, extremely limited guidance is available on routine canine decontamination from pathogenic biological materials. In this article, we evaluate the ability of several commercial off-the-shelf cleansing products, used in wipe form, to remove superficial contamination from fur, canine equipment, and toys. Using Glo Germ MIST as a proxy for biological contamination, our analysis demonstrated more than a 90% average reduction in contamination after wiping with a Nolvasan scrub solution, 0.5% chlorhexidine solution, or 70% isopropyl alcohol. Wiping with nondisinfectant baby wipes or water yielded an almost 80% average removal of contaminant from all surfaces. Additionally, researchers used Gwet's AC2 measurement to assess interrater reliability, which demonstrated substantial agreement (P < .001). These data provide key insights toward the development of a rapid, convenient, and fieldable alternative to traditional water-intensive bathing of working canines.

Complete Genome Sequence of Francisella tularensis Live Vaccine Strain NR-28537 (BEI Master Cell Bank).

The genome of Francisella tularensis live vaccine strain NR-28537 was sequenced by a hybrid approach utilizing an Oxford Nanopore Technologies R9 flow cell and an Illumina MiSeq platform. De novo assembly of the resulting long and short reads produced a single-contig whole-genome sequence.

Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo.

The 2014-2016 Zika virus (ZIKV) epidemic in the Americas resulted in large deposits of next-generation sequencing data from clinical samples. This resource was mined to identify emerging mutations and trends in mutations as the outbreak progressed over time. Information on transmission dynamics, prevalence, and persistence of intra-host mutants, and the position of a mutation on a protein were then used to prioritize 544 reported mutations based on their ability to impact ZIKV phenotype. Using this criteria, six mutants (representing naturally occurring mutations) were generated as synthetic infectious clones using a 2015 Puerto Rican epidemic strain PRVABC59 as the parental backbone. The phenotypes of these naturally occurring variants were examined using both cell culture and murine model systems. Mutants had distinct phenotypes, including changes in replication rate, embryo death, and decreased head size. In particular, a NS2B mutant previously detected during in vivo studies in rhesus macaques was found to cause lethal infections in adult mice, abortions in pregnant females, and increased viral genome copies in both brain tissue and blood of female mice. Additionally, mutants with changes in the region of NS3 that interfaces with NS5 during replication displayed reduced replication in the blood of adult mice. This analytical pathway, integrating both bioinformatic and wet lab experiments, provides a foundation for understanding how naturally occurring single mutations affect disease outcome and can be used to predict the of severity of future ZIKV outbreaks. To determine if naturally occurring individual mutations in the Zika virus epidemic genotype affect viral virulence or replication rate in vitro or in vivo, we generated an infectious clone representing the epidemic genotype of stain Puerto Rico, 2015. Using this clone, six mutants were created by changing nucleotides in the genome to cause one to two amino acid substitutions in the encoded proteins. The six mutants we generated represent mutations that differentiated the early epidemic genotype from genotypes that were either ancestral or that occurred later in the epidemic. We assayed each mutant for changes in growth rate, and for virulence in adult mice and pregnant mice. Three of the mutants caused catastrophic embryo effects including increased embryonic death or significant decrease in head diameter. Three other mutants that had mutations in a genome region associated with replication resulted in changes in in vitro and in vivo replication rates. These results illustrate the potential impact of individual mutations in viral phenotype.

Changing student attitudes and perceptions toward opioid use disorder.

INTRODUCTION: With the opioid epidemic creating a group of patients with unique health care needs, pharmacists have an opportunity to be a good resource for patients recovering from opioid use disorder (OUD). To accomplish this, it is essential that pharmacists are knowledgeable and unbiased toward this patient population. METHODS: Because the curriculum in place to obtain a PharmD at Drake University does not include in-depth information on substance use disorders, study investigators offered students an opportunity to receive more intensive education. Faculty members at Drake University provided didactic and panel discussion presentations on topics such as opioid pharmacology, OUD, and treatment options. The students were assessed for their perception of knowledge and stigma before and after the summit by using a 5-point Likert scale to measure their attitudes toward 10 statements. RESULTS: Total knowledge scores showed a significant change of 3.1, indicating an increase in perceived understanding of materials presented (P < .0001). Total stigma scores also changed by 1.4, illustrating a statistically significant decrease in negative perceptions (P = .0198). DISCUSSION: By providing more in-depth education, the summit showed that increasing pharmacy student knowledge about OUD and its treatment may decrease associated stigma.

Proposed model for translational research at a teaching-intensive college of pharmacy.

BACKGROUND: Many American colleges of pharmacy are small, private, teaching institutions. Faculty are required to maintain a research agenda, although the publication quota is less compared with their publicly funded college of pharmacy peers. Faculty at these smaller schools conduct research with very little internal or external funding. This tends to lead to smaller, less impactful research findings. Translational research is becoming popular for research faculty as it bridges theory to practice. The Knowledge-to-Action (KTA) framework presents the steps to conduct translational research. PURPOSE: To apply and determine if the KTA framework would be able to produce practice-impactful research at an institution that does not depend on grant funding as part of faculty research agendas. PROCEDURES: An interdisciplinary team was formed with providers at the clinical faculty's practice site. As the team moved through the KTA steps, authors documented the roles of each team member. It was clear that many different types of teams were formed throughout the KTA process. These teams were then categorized according to the Interdisciplinary Teamwork System. The final result is a proposed model of types of teams and required member roles that are necessary within each KTA step for faculty to conduct practice-impactful research at a small, private, teaching institution without substantial grant funding awards. MAIN FINDINGS: Applying the KTA framework, two impactful original research manuscripts were developed over two academic years. Furthermore, the practitioners at the clinical faculty member's site were very pleased with the ease of conducting research, as they were never required to take a lead role. In addition, both faculty members alternated lead and support role allowing for a decreased burden of workload while producing theory-driven research. CONCLUSION: The KTA framework can create a model for translational research and may be particularly beneficial to small teaching institutions to conduct impactful research.