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To identify how patient journey mapping is being undertaken and reported.A scoping review of the literature was undertaken using JBI guidance.Databases were searched in July 2021 (16th-21st), including Ovid's Medline, Embase, Emcare and PsycINFO; Scopus; Web of Science Core Collection, the Directory of Open Access Journals; Informit and; ProQuest Dissertations and Theses Global.Eligible articles included peer-reviewed literature documenting journey mapping methodologies and studies conducted in healthcare services. Reviewers used Covidence to screen titles and abstracts of located sources, and to screen full-text articles. A table was used to extract data and synthesize results.Eighty-one articles were included. An acceleration of patient journey mapping research was observed, with 76.5% (n = 62) of articles published since 2015. Diverse mapping approaches were identified. Reporting of studies was inconsistent and largely non-adherent with relevant, established reporting guidelines.Patient journey mapping is a relatively novel approach for understanding patient experiences and is increasingly being adopted. There is variation in process details reported. Considerations for improving reporting standards are provided.Patient journey mapping is a rapidly growing approach for better understanding how people enter, experience and exit health services. This type of methodology has significant potential to inform new, patient centred models of care and facilitate clinicians, patients and health professionals to better understand gaps and strategies in health services. The synthesised results of this review alert researchers to options available for journey mapping research and provide preliminary guidance for elevating reporting quality.

Objective This scoping review will assess the literature that documents or utilizes patient journey mapping methodologies in health care settings. It will also examine the reporting processes of studies that use this methodology. Introduction Health care systems are complex and can be challenging for patients to navigate. Using patient journey mapping as a research method promotes a deeper understanding of patient experiences when navigating these systems. Patient journey mapping provides valuable insights into where systems are working well, where gaps in care exist, and how the system could respond to these gaps. Inclusion criteria This review will consider peer-reviewed articles and publicly available academic literature documenting patient journey mapping methodologies. The review will also consider studies providing guidance and recommendations on how to report patient journey mapping studies in health care services and systems. Methods The proposed review will follow JBI guidance for scoping reviews. The following databases will be searched: MEDLINE, Embase, Emcare, PsycINFO, Scopus, Web of Science Core Collection, the Directory of Open Access Journals, Informit, and ProQuest Dissertations and Theses Global. The search will not be limited to year of publication but will be limited to studies reported in English. The PRISMA-ScR extension will be used to document the literature search. Two reviewers will screen titles, abstracts, and full-text articles. An extraction table will be used to extract relevant data from all included articles and to facilitate data analysis.

Deregulated polyamine biosynthesis is emerging as a common feature of neuroblastoma and drugs targeting this metabolic pathway such as DFMO are in clinical and preclinical development. The polyamine analog verlindamycin inhibits the polyamine biosynthesis pathway enzymes SMOX and PAOX, as well as the histone demethylase LSD1. Based on our previous research in acute myeloid leukemia (AML), we reasoned verlindamycin may also unblock neuroblastoma differentiation when combined with all-trans-retinoic acid (ATRA). Indeed, co-treatment with verlindamycin and ATRA strongly induced differentiation regardless of MYCN status, but in MYCN-expressing cells, protein levels were strongly diminished. This process was not transcriptionally regulated but was due to increased degradation of MYCN protein, at least in part via ubiquitin-independent, proteasome-dependent destruction. Here we report that verlindamycin effectively induces the expression of functional tumor suppressor—antizyme via ribosomal frameshifting. Consistent with previous results describing the function of antizyme, we found that verlindamycin treatment led to the selective targeting of ornithine decarboxylase (the rate-limiting enzyme for polyamine biosynthesis) as well as key oncoproteins, such as cyclin D and Aurora A kinase. Retinoid-based multimodal differentiation therapy is one of the few interventions that extends relapse-free survival in MYCN-associated high-risk neuroblastoma and these results point toward the potential use of verlindamycin in this regimen.

At the onset of the COVID-19 global pandemic, Florida’s State Emergency Response Team’s Emergency Support Function 8 (ESF-8) Health and Medical Staffing Unit faced a surge of personnel requests from the field. The unit found that, given the scope of requests, standard disaster staffing practices could not always accommodate the requirements of the requests. With full support of leadership, the ESF-8 Staffing Unit developed new and innovative practices to streamline the cumbersome hiring process including coordinating with internal and external partners to expedite staff identification and implementing just-in-time training.

User consultation is an essential first step in assuring high-quality flow cytometric data. A central challenge to shared resource laboratory (SRL) staff is how to best guide new and current users to meet each projects' needs. One solution to this challenge is to follow a standard user consultation platform addressing all critical steps between the conception of the experiment and the actual acquisition of samples. Here we describe considerations to help an SRL understand the researcher's goals and how best the SRL staff can provide expert advice in a structured manner. User consultation has an educational nature, informing users about current best practices in cytometry that apply to their specific utilization. A consultation report also improves communication between the SRL, principal investigator, and lab members of the collaborating researcher. Development of best SRL practices is spearheaded by the ISAC SRL committee and this communication sets the foundation to initiate such report for user consultation. Implementation of best practices during user consultation will improve rigor and reproducibility in cytometry.

Background\ud Prostate cancer (PC) tissue contains all‐trans retinoic acid (ATRA) at a very low level (10−9 M), at least an order of magnitude lower than in adjacent normal healthy prostate cells or benign prostate hyperplasia. When this is coupled with deregulated expression of the intracellular lipid‐binding proteins FABP5 and CRABP2 that is frequently found in PC, this is likely to result in the preferential delivery of ATRA to oncogenic PPARβ/δ rather than retinoic acid receptors (RARs). There are three isotypes of RARs (RARα, RARβ, and RARγ) and recent studies have revealed discrete physiological roles. For example, RARα and RARγ promote differentiation and self‐renewal, respectively, which are critical for proper hematopoiesis.\ud \ud Aims\ud We have previously shown that ATRA stimulates transactivation of RARγ at sub‐nanomolar concentrations (EC50 0.24 nM), whereas an 80‐fold higher concentration was required for RARα‐mediated transactivation (EC50 19.3 nM). Additionally, we have shown that RAR pan‐antagonists inhibit the growth of PC cells (at 16‐34 nM). These findings, together with the low level of ATRA in PC, led us to hypothesize that RARγ plays a role in PC pathogenesis and that RARγ‐selective antagonism may be an effective treatment.\ud \ud Methods and results\ud We found that concentrations of 10−9 M and below of ATRA promoted survival/proliferation and opposed adipogenic differentiation of human PC cell lines by a mechanism that involves RARγ. We also found that a RARγ‐selective antagonist (AGN205728) potently induced mitochondria‐dependent, but caspase‐independent, cell death in PC cell lines. Furthermore, AGN205728 demonstrated synergism in killing PC cells in combination with cytotoxic chemotherapeutic agents.\ud \ud Conclusion\ud We suggest that the use of RARγ‐selective antagonists may be effective in PC (and potentially other cancers), either as a single agent or in combination with cytotoxic chemotherapy.

Background: Evidence shows systems change interventions improve care and outcomes for stroke patients. Geopolitical boundaries have been a barrier to improving regional systems of care. Despite efforts nationally, regionally, and locally alteplase use for ischemic stroke has remained low and door to needle (DTN) times exceeded 60 minutes. Kentucky created the Stroke Encounter Quality Improvement Project (SEQIP) in 2009 to share best practices and improve stroke systems of care across the Commonwealth. Purpose: The aim was to utilize and share best practice models among 23 SEQIP hospitals in KY to improve tPA utilization, decrease DTN times, and improve outcomes. Methods: Hospitals implemented a statewide quality improvement plan focused on identifying barriers, removing barriers, and implementing best practice strategies regarding thrombolytic therapy. Accountability was achieved with ongoing GWTG data tracking, teleconferences, and face to face meetings from January 2009 through December 2018 sharing strategies and solutions for best practice. Results: SEQIP’s participating hospitals achieved significant improvement in thrombolytic administration over 10 years. The percent of all AIS patients receiving tPA increased from 4.61% in 2009 to 8.80% in 2018 (OR=2.0, p Conclusions: Geopolitical boundaries can be overcome and collaboration can be sustained among competing hospitals through sharing of best practices to safely increase utilization of tPA in eligible patients, decrease DTN times, and improve outcomes.

Background: The Stroke Encounter Quality Improvement Project (SEQIP) launched in 2009 as a statewide voluntary initiative and collaboration between the American Heart Association, the Kentucky Department for Public Health and 16 acute care hospitals interested in improving stroke care in their communities. The mission is to advance acute stroke care and reduce disparities in Kentucky by: establishing a network that encourages and supports collaboration; increases access to stroke care by targeting underserved areas; provides opportunities to share resources related to program development and proficiency across the continuum of care; and promotes quality outcomes and standardization of care through collegiality and use of evidence-based guidelines and research collaboration. Purpose: The goal of this unfunded initiative (now in its tenth year) has been to increase adherence to evidence-based guidelines for stroke patients by implementing a unified statewide effort. Methods: Get With The Guidelines-Stroke data were reviewed with the founding 16 SEQIP hospitals and adherence to evidence-based guidelines was measured and analyzed over a 10-year period. Results: SEQIP has grown to a network of 35 hospitals with 23 submitting data; patient records increased from 4358 (2008) to 10015 (2018); hospitals achieving Gold GWTG award status increased from 4 to 16; certified stroke centers grew from 4 to 32 decreasing geographic barriers to the nearest certified center; and, SEQIP hospitals achieved statistically significant improvement in all stroke measures. Conclusions: With deployment of strategically targeted action plans and expected accountability, competing hospitals can collaborate on a statewide level. Sharing of best practices across organizations can empower stroke teams to implement effective strategies within the confines of their resources to achieve collective goals.

Although there is a long history of research in the area of Academic Literacies in Brazil, Ferreira e Stella (2018, p. 17) affirm that studies on learning and teaching writing for academic purposes in Brazilian universities is still a neglected action. Considering the increased exigence to publish internationally and the need to ground pedagogical practices for writing in results from research in each of the different disciplinary contexts (MOTTA-ROTH, 2013, p. 12), this study aims to explore academic literacy practices in English and their development in a highly-ranked graduate program that presents high productivity in international publications in English, the Graduate Program in Chemistry at UFSM. Toward that end, we propose the following research questions: 1) which academic literacy practices are most valued by the professors, both in terms of their own practices and those of their students, and why? 2) how did the professors develop their own academic literacies and which aspects present the greatest level of difficulty for them? 3) what dynamics of coauthorship and/or pedagogic approaches do the professors adopt and which aspects of writing present the greatest level of difficulty for their students? The interdisciplinary theoretical framework combines Genre Studies (BAZERMAN, 2007; BAWARSHI; REIFF, 2013), Academic Literacies (LEA; STREET 1998, IVANIC 1998) and Critical Discourse Analysis (FAIRCLOUGH, 2003), as well as the concepts of Communities of Practice and Legitimate Peripheral Participation (LPP) (LAVE; WENGER, 1991; WENGER, 1998). Using data generated from a semistructured questionnaire and qualitative interviews, we analyzed the discourses of the professors, exploring whether the development of skills, competencies and discursive conditions involved in academic literacies in English is mediated by LPP in a community structured to provide increasing access to new members and capacitate them to participate fully in the community. We argue that the community studied demonstrates variable degrees of student LPP. Professors who report a greater degree of LPP along with a network for collaborating on writing and revising among the students also report less difficulty in the development of the students’ academic literacies. On the other hand, even when a greater degree of LPP is observed, we detect the need for pedagogical approaches that could supplement the existing LPP. The professors’ discourse reveals a dialectical tension, where, on the one hand, scientific writing is portrayed as easy and, on the other hand, writing articles is characterized as difficult and the professors report that students encounter great difficulty. We argue that this tension is related, in most of the accounts, to a pedagogical approach in line with the notion of academic literacies as cognitive skills acquired through tacit knowledge, rather than through explicit teaching within a perspective of academic literacies as social practices. In this sense, we recommend the creation of collaborative networks in the community and the introduction of a collaborative and transformative pedagogical approach that will allow students to identify as authors within the community, viewing academic literacies as social actions and developing a critical eye toward the texts and genre systems that constitute them. Embora tenha uma longa história de produção de conhecimento na área de Letramentos Acadêmicos (LA) no Brasil, Ferreira e Stella (2018, p. 17) apontam que a produção de pesquisas em universidades brasileiras sobre o ensino e aprendizagem de escrita para fins acadêmicos ainda é uma ação negligenciada. Considerando o aumento na cobrança para publicações e internacionalização e a necessidade de fundamentar práticas pedagógicas para a produção textual em resultados de pesquisa em cada um dos diferentes contextos disciplinares (MOTTA-ROTH, 2013, p. 12), este estudo visa explorar as práticas de letramento acadêmico (LA) em inglês e sua aprendizagem em um programa de pós-graduação bem-conceituado que apresenta alta produtividade de publicação internacional em inglês, o Programa de Pós-graduação em Química da UFSM (PPGQ). Para esse objetivo, propomos as seguintes perguntas de pesquisa: 1) Quais práticas de LA em inglês são mais valorizadas pelos professores, tanto para sua própria produção como para a produção de seus estudantes, e por quê? 2) Como os professores desenvolveram seus LA em inglês e quais aspectos apresentam maior grau de dificuldade para eles? 3) Quais dinâmicas de coautoria e/ou abordagens pedagógicas utilizam para o desenvolvimento dos LA de seus estudantes e em quais aspectos de escrita os estudantes apresentam maior grau de dificuldade? O quadro teórico adotado é interdisciplinar ao combinar a Sociorretórica (BAZERMAN, 2007; BAWARSHI; REIFF, 2013), os LA (LEA; STREET 1998, IVANIC 1998) e a Análise Crítica do Discurso (ACD) (FAIRCLOUGH, 2003). Também são tomadas como referências os conceitos de Comunidades de Prática (CdP) e Participação Periférica Legítima (PPL) (LAVE; WENGER, 1991; WENGER, 1998). Usando dados gerados num questionário semiestruturado e entrevistas qualitativas, analisamos criticamente os discursos dos professores, explorando se a aprendizagem de habilidades, competências e condições discursivas envolvidas nos LA em inglês é mediada pela PPL em uma comunidade estruturada para dar acesso crescente a novos membros e formá-los para participar de forma plena na comunidade. Argumentamos que a CdP estudada demonstra graus variáveis de PPL dos estudantes. Professores que relatam um maior grau de PPL junto a uma rede colaborativa para escrita e revisão entre seus alunos também relatam menos dificuldades no desenvolvimento dos LA de seus alunos. Por outro lado, mesmo quando exista maior grau de PPL, constatamos a necessidade de abordagens pedagógicas capazes de subsidiar a PPL existente. O discurso dos professores revela uma tensão dialética em que, por um lado, a escrita científica é caracterizada como fácil e, por outro lado, a escrita de artigos é retratada como difícil e os professores reportam que seus alunos apresentam muitas dificuldades. Argumentamos que essa tensão está ligada, na maior parte dos relatos, a uma abordagem pedagógica alinhada com a noção de os LA serem habilidades cognitivas adquiridas por meio de conhecimento tácito e não pelo ensino explícito dentro de uma perspectiva de LA como práticas sociais. Nesse sentido, recomendamos a criação de redes colaborativas na CdP e a introdução de uma abordagem pedagógica transformativa e colaborativa que permita que o aluno se identifique como autor dentro da comunidade, compreendendo os LA como ações sociais e desenvolvendo um olhar crítico em relação aos textos e sistemas de gênero que os constituem.

Background: The Stroke Encounter Quality Improvement Project (SEQIP) is a collaboration between certified stroke centers, the AHA/ASA and the Kentucky Department for Public Health (KDPH) to implement statewide QI initiatives to improve the care of stroke patients. From 2009 to 2018, 23 hospital in Kentucky participating in SEQIP have entered 76,222 stroke patient records into Get With The Guidelines® (GWTG) / Patient Management Tool™ (PMT). Purpose: Geographic information systems (GIS) tools can expand our understanding of care and outcomes based on patient location. The purpose of this project was to demonstrate the methods of linking a disease management registry with GIS mapping and analysis program,) to understand challenges when performing this link, and to derive meaningful insight on stroke care and outcomes by zip code. Methods: Stroke data from GWTG and PMT was compiled and downloaded by KDPH. The information was converted to a database file for use in ArcGIS. After excluding those who had missing or incomplete zip codes, records were Geocoded annually from 2009 to 2018. The data were then matched to one of 945 zip codes in Kentucky. Data were summarized by zip code, calendar year by the number of ischemic strokes; number IV alteplase administration; rate ischemic stroke receiving IV alteplase; number and rate of ischemic stroke patients arriving to hospital by EMS, privately owned vehicle or transfer; and median time from last known well to hospital arrival; and medical history of hypertension. Additional data including hospitals, certified stroke centers, drive time analysis, etc. were added to maps. Results: Mapping GWTG and PMT stroke data is feasible and may allow for additional analysis by location. Conclusion: Using GIS mapping and methodology can assist hospital stroke coordinators and public health officials in developing and implementing interventions to improvement stroke care and outcomes. Further analysis including socioeconomic, demographic and marketing/consumer preference data is planned to better understand variations by zip codes. This feasibility project provides an example of a useful application of GIS analyses applied to data registry including GWTG and PMT.

71 Background: With the advent of newer non-cytotoxic therapies for aPC, early integration of hormonal therapies (HT), and expansion of services within urology practices [e.g., radiation therapy (RT) and immunotherapy], urology practices are playing a larger role in the management of patients with aPC. In this descriptive study, we sought to assess the differences between medical oncologists (oncs) and urologists (uros) as they pertain to referral patterns and treatment decisions of men with aPC. Methods: Virtual meetings held in August 2020 convened oncs and uros of diverse US regions and practice types with experience treating prostate cancer (PC) to better understand perceptions around the management of patients with PC. Participants submitted responses via web-based pre-meeting surveys and real-time polling. All responses are summarized using descriptive statistics. Results: 66 oncs and 69 uros participated. The advisors were mostly community-based and see on average 20+ patients per day. 75% of uros estimated they refer ≤25% of all of their patients with PC to oncs and 75% of oncs reported that > 50% of their patients with PC referrals are from uros. Other referring physicians identified include primary care (76%), radiation oncs (46%), or hospitalists (40%). Uros perceive oncs as co-managers (86%) for their patients with aPC and rarely (9%) transfer the care of their patients completely to oncs. Referrals from uros to oncs are driven by the need for chemotherapy (chemo) (52%) or progression to metastatic castration-resistant PC (mCRPC) (22%). Oncs reported that upon referral, these patients with PC have already been exposed to HT (75%), RT (66%), and/or surgery (43%). For second-line treatment of asymptomatic patients with mCRPC, oncs most commonly prescribe HT (60%) and chemo (23%), while uros most commonly prescribe sipuleucel-T (45%) and HT (38%). For second-line treatment of patients with mCRPC and symptomatic bony disease, oncs most commonly prescribe chemo (44%) and radium-223 (34%), while uros most commonly prescribe radium-223 (47%) and HT (22%). Conclusions: Uros refer patients with aPC to oncs for chemo. Uros prefer to treat with non-chemo options when possible and retain oncologic care for most of the patients’ cancer journey. The need for chemo is a major reason for referral from uros to oncs. The impact of these provider preferences upon patient outcomes in the real world needs further research. [Table: see text]

Influenza A virus (IAV) is a major respiratory pathogen of both humans and animals. The lung is protected from pathogens by alveolar epithelial cells, tissue-resident alveolar macrophages, dendritic cells, and mast cells. The role of alveolar epithelial cells, endothelial cells, and alveolar macrophages during IAV infection has been studied previously. In this study, we address the role of mast cells during IAV infection. Respiratory infection with A/WSN/33 causes significant disease and immunopathology in C57BL/6 mice but not in B6.Cg-KitW-sh mice, which lack mast cells. During in vitro coculture, A/WSN/33 caused mast cells to release histamine, secrete cytokines and chemokines, and produce leukotrienes. Moreover, when mast cells were infected with IAV, the virus did not replicate within mast cells. Importantly, human H1N1, H3N2, and influenza B virus isolates also could activate mast cells in vitro. Mast cell production of cytokines and chemokines occurs in a RIG-I/MAVS–dependent mechanism; in contrast, histamine production occurred through a RIG-I/MAVS–independent mechanism. Our data highlight that, following IAV infection, the response of mast cells is controlled by multiple receptors. In conclusion, we identified a unique inflammatory cascade activated during IAV infection that could potentially be targeted to limit morbidity following IAV infection.

Influenza A virus (IAV) is a widespread infectious agent commonly found in mammalian and avian species. In humans, IAV is a respiratory pathogen that causes seasonal infections associated with significant morbidity in young and elderly populations, and has a large economic impact. Moreover, IAV has the potential to cause both zoonotic spillover infection and global pandemics, which have significantly greater morbidity and mortality across all ages. The pathology associated with these pandemic and spillover infections appear to be the result of an excessive inflammatory response leading to severe lung damage, which likely predisposes the lungs for secondary bacterial infections. The lung is protected from pathogens by alveolar epithelial cells, endothelial cells, tissue resident alveolar macrophages, dendritic cells, and mast cells. The importance of mast cells during bacterial and parasitic infections has been extensively studied; yet, the role of these hematopoietic cells during viral infections is only beginning to emerge. Recently, it has been shown that mast cells can be directly activated in response to IAV, releasing mediators such histamine, proteases, leukotrienes, inflammatory cytokines, and antiviral chemokines, which participate in the excessive inflammatory and pathological response observed during IAV infections. In this review, we will examine the relationship between mast cells and IAV, and discuss the role of mast cells as a potential drug target during highly pathological IAV infections. Finally, we proposed an emerging role for mast cells in other viral infections associated with significant host pathology.

The efficiency and effectiveness of document delivery systems used within a library network can be a strong influence on faculty and student attitudes toward resource sharing. This article describes a survey administered by one library consortium to investigate usage of a courier service known as ABC Express, a document delivery service that delivers requested items among three libraries three times a week. Issues such as awareness, delivery speed, fill rate, and e-mail notification are explored. Decisions of what items to own and access can be made in light of available data in conjunction with a better knowledge of user needs and concerns.

Dave Richley and Amy Graham argue that smartphones may provide an innovative solution to capturing ECG when palpitations are too infrequent for conventional recording

Background. There is a significant resumption of smoking following smoking cessation using varenicline. Both smoking cessation medications and counseling have been shown to increase smoking quit rates at one year. Thus, the combination of varenicline and interactive voice response (IVR) telephony followed by extended IVR may further improve smoking cessation rates at one and two years. Methods 101 participants were recruited from the community via newspaper advertisement. They attended a group counseling session and were given smoking information booklets from the Canadian Cancer Society. After 12 weeks of varenicline and 9 IVR calls, all participants who had quit smoking were randomized into 2 groups matched by levels of motivation and addiction as per baseline questionnaire score. The intervention group continued to receive bi-weekly IVR support for weeks 13 – 52. The control group no longer received IVR. The primary end-point was self-reported abstinence and exhaled carbon monoxide levels of less than 10 ppm for weeks 12, 52 and 2 years. Data were analyzed by Fisher’s exact test or Wilcoxon rank-sum test. Results Of the 101 participants, 44 (43%) had stopped smoking after 12 weeks of varenicline and 9 IVR calls. Of these, 23 (52%) were randomized to receive IVR calls from weeks 13 to 52. At 52 weeks, 26 (59%) participants remained smoke-free. Of the 23 with IVR, 12 (52.2%) stopped smoking compared to 14 of 21 (66.7%) without IVR. At 2 years, 40 of the 44 (90.9%) randomized participants were contacted and 24 of the 44 (54.5%) came in for testing. Fourteen (13% of the original cohort, 30% who were abstinent at 12 weeks and 53% who were abstinent at 52 weeks) remained smoke-free. Five of the 23 (21.7%) randomized to IVR and 9 of the 21 (42.9%) randomized to no IVR remained smoke-free at 2 years. Conclusions In this pilot study of an apparently healthy population, extended IVR did not affect abstinence rates. There was no relapse prevention benefit in offering 9 months of continued IVR to subjects who had stopped smoking after receiving 3 months of varenicline and IVR treatment. Trial registration ClinicalTrial.gov: NCT00832806

Influenza A virus (IAV), a seasonal respiratory pathogen, results in severe lung pathology. Recent global analysis of lungs from mice infected with highly pathological IAV strains demonstrated enrichment of mast cells, but the role of mast cells during severe pulmonary viral infections has been under studied. We have recently shown that A/WSN/33 causes significant immunopathology in C57BL/6 mice and is mast cell-dependent. A/WSN/33(WSN) was able to directly activate mast cells to produce histamine, leukotrienes, inflammatory cytokines, and anti-viral chemokines. IAV-induced cytokine storm is thought to induce viral immunopathology, therefore we explored the importance of this response from mast cells. Mast cell cytokine production occurred in a RIG-I/MAVS-dependent fashion, but reconstitution of mast cell-deficient mice with RIG-I-/- mast cells generated lung pathology similar to wild type mast cells. Thus, it appears that mast cell degranulation rather than production of cytokines causes WSN-induced lung pathology. Mast cell degranulation occurs early in the viral entry cycle through a RIG-I/MAVS- and MyD88/Trif-independent mechanism. Using recombinant WSN strains, we found an association between the WSN hemaggultinin and neuraminidase proteins in the activation of mast cell degranulation and WSN-induced disease. We have identified a unique inflammatory cascade which could be therapeutically targeted to limit morbidity following infection with influenza virus.

Influenza A virus (IAV), a seasonal respiratory pathogen, has a high capacity to cause pandemic infections resulting in severe lung pathology. Multiple studies using pathological IAV strains have demonstrated the importance of macrophages and neutrophils, but whether the pathological inflammation induced by different IAV isolates is identical has not been directly compared. Here we demonstrate that while both A/WSN/33 (WSN) and A/PR/8/34 (PR8) influenza viruses cause significant immunopathology in mice the inflammatory milieu differs between the two isolates. Specifically, PR8 infection was associated with greater neutrophil recruitment and CCL4 and CXCL2 expression; in contrast, WSN infection was associated mast cell activation, recruitment of a unique Ly6ghigh Ly6chigh monocyte population, and elevated levels of IFNα, G-CSF, and CCL2. One difference between WSN and PR8 is their hemaggultinin molecules preferentially bind to α2,6- and α2,3-linked sialic acids, respectively. Using a recombinant WSN strain whose sialic acid binding preference is switched to α2,3-linkages, we observed minimal mast cell activation and a switch to a more neutrophilic inflammation. Thus, sialic acid binding preference of IAV isolates has clinically important alterations to the inflammatory cascade(s) activated. These findings have important implications for our ability to therapeutically target the host inflammatory response to limit morbidity following influenza virus infection

Influenza A virus (IAV) is a seasonal respiratory pathogen that can result in severe lung pathology. The role of mast cells has been under explored during severe pulmonary viral infections. We found that both A/WSN/33 (WSN) and A/PR/8/34 (PR8) influenza viruses cause significant immunopathology in C57BL/6 mice, but only WSN induced pathology was mast cell dependent. Using in vitro-derived bone marrow cultured mast cells (BMCMC), we found that WSN, but not PR8, directly activated BMCMC to produce histamine, leukotrienes, inflammatory cytokines, and anti-viral chemokines. Moreover, human H1N1, H3N2, and influenza B virus isolates could activate both murine BMCMC and the human mast cell line HMC-1 in vitro. BMCMC activation required infection of mast cells by IAV, which was dependent on the viral hemagglutinin. Cytokine and chemokine production from BMCMC occurs in a RIG-I/MAVS-dependent fashion which required the de novo production of vRNA; conversely, degranulation occurs through a RIG-I/MAVS-independent mechanism. Reconstitution of mast cell deficient mice with RIG-I-/- BMCMC generated lung pathology similar to wild type BMCMC, suggesting that mast cell degranulation, rather than production of cytokines, causes WSN induced lung pathology. Thus, we have identified a unique inflammatory cascade which could be therapeutically targeted to limit morbidity following infection with influenza virus.

The lung is protected from pathogens by alveolar epithelial cells, tissue resident alveolar macrophages, dendritic cells, and mast cells. The role of the mast cell has been under explored during severe pulmonary viral infections. In mice that lack functional mast cells, pathological immune responses are altered depending on the influenza strain. We found that both A/WSN/33 (WSN) and A/PR/8/34 (PR8) influenza viruses generate significant immunopathology in C57BL/6 mice, but only WSN induced pathology was mast cell dependent. To assess the differences in mast cell dependent inflammation response due to influenza strain, we examined the ability of each strain to activate in vitro-derived bone marrow cultured mast cells (BMCMC). We found that WSN, but not PR8, influenza virus activated BMCMC to produce inflammatory cytokines/chemokines, anti-viral chemokines, and degranulate. Importantly, a recombinant PR8 virus that expresses the WSN hemagglutinin (HA) glycoprotein instead of its own could induce cytokine production in BMCMC, demonstrating an important role for the HA glycoprotein for mast cell activation. Mast cell infection and viral RNA synthesis and export were necessary for mast cell activation and occurred in a MAVS-dependent mechanism. Thus, we have identified a unique inflammatory cascade which could be targeted to limit morbidity following infection with influenza virus.

Understanding the innate inflammatory pathways activated during influenza A virus infection is vital to developing effective therapeutics. Our results demonstrate that the inflammatory cascades induced by different influenza A virus strains are not identical. Mice infected with A/WSN/33 show less airway hyper-reactivity than mice infected with A/PR/8/34, likely due to decreased expression of the IL-33/IL-13 axis of inflammation. In addition, we have examined the lipidomic expression profile during flu infections from each of these strains. Lipid extracts were prepared from whole lungs, and specific lipid mediators were both identified and quantified using UHPLC-MS. This approach provides an automated, high-throughput, and high-sensitivity technique to characterize the lipidomic fingerprint of the infection resulting from various influenza A strains. The compounds differentially expressed between strains will ultimately be used to develop targeted therapeutics to lessen both the morbidity and mortality of the seasonal flu.