bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autographa californicaNucleopolyhedrovirus Ac76: a Dimeric Type II Integral Membrane Protein That Contains an Inner Nuclear Membrane-Sorting Motif
Authors: Wei Denghui , Wang Yan , Zhang Xiaomei , Hu Zhaoyang , Yuan Meijin , Yang Kai 
Published: November 2013

ABSTRACT

Our previous study showed that the
Autographa californica
Nucleopolyhedrovirus
(AcMNPV)
ac76
gene is essential for both budded virion (BV) and occlusion-derived virion (ODV) development. More importantly, deletion of
ac76
affects intranuclear microvesicle formation. However, the exact role by which
ac76
affects virion morphogenesis remains unknown. In this report, we characterized the expression, distribution, and topology of Ac76 to further understand the functional role of Ac76 in virion morphogenesis. Ac76 contains an α-helical transmembrane domain, and phase separation showed that it was an integral membrane protein. In AcMNPV-infected cells, Ac76 was detected as a stable dimer that was resistant to SDS and thermal denaturation, and only a trace amount of monomer was detected. A coimmunoprecipitation assay demonstrated the dimerization of Ac76 by high-affinity self-association. Western blot analyses of purified virions and their nucleocapsid and envelope fractions showed that Ac76 was associated with the envelope fractions of both BVs and ODVs. Immunoelectron microscopy revealed that Ac76 was localized to the plasma membrane, endoplasmic reticulum (ER), nuclear membrane, intranuclear microvesicles, and ODV envelope. Amino acids 15 to 48 of Ac76 were identified as an atypical inner nuclear membrane-sorting motif because it was sufficient to target fusion proteins to the ER and nuclear membrane in the absence of viral infection and to the intranuclear microvesicles and ODV envelope during infection. Topology analysis of Ac76 by selective permeabilization showed that Ac76 was a type II integral membrane protein with an N terminus exposed to the cytosol and a C terminus hidden in the ER lumen.
Inhibition of NF-κB-Mediated Inflammation in Severe Acute Respiratory Syndrome Coronavirus-Infected Mice Increases Survival
Authors: DeDiego Marta L. , Nieto-Torres Jose L. , Regla-Nava Jose A. , Jimenez-Guardeño Jose M. , Fernandez-Delgado Raul , Fett Craig , Castaño-Rodriguez Carlos , Perlman Stanley , Enjuanes Luis 
Published: November 2013

ABSTRACTSevere acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of a respiratory disease that has a 10% mortality rate. We previously showed that SARS-CoV lacking the E gene (SARS-CoV-ΔE) is attenuated in several animal model systems. Here, we show that absence of the E protein resulted in reduced expression of proinflammatory cytokines, decreased numbers of neutrophils in lung infiltrates, diminished lung pathology, and increased mouse survival, suggesting that lung inflammation contributed to SARS-CoV virulence. Further, infection with SARS-CoV-ΔE resulted in decreased activation of NF-κB compared to levels for the wild-type virus. Most important, treatment with drugs that inhibited NF-κB activation led to a reduction in inflammation and lung pathology in both SARS-CoV-infected cultured cells and mice and significantly increased mouse survival after SARS-CoV infection. These data indicated that activation of the NF-κB signaling pathway represents a major contribution to the inflammation induced after SARS-CoV infection and that NF-κB inhibitors are promising antivirals in infections caused by SARS-CoV and potentially other pathogenic human coronaviruses.
Adenovirus Detection by the cGAS/STING/TBK1 DNA Sensing Cascade
Authors: Lam Eric , Stein Saskia , Falck-Pedersen Erik 
Published: November 2013

ABSTRACT

Adenovirus (Ad) infection triggers a cell-specific antiviral response following exposure of viral DNA to the intracellular compartment. A variety of DNA sensors (DAI, AIM2, DDx41, RNA polymerase [Pol] III, and IFI16 [p204]) have been identified in recent years; however, the DNA sensor involved in detection of adenovirus has not been established. Cyclic GMP-AMP synthase (cGAS), a DNA sensor that produces a cyclic guanine-adenine dinucleotide (cGAMP) inducer of STING, has been examined to determine its role in generating an antiadenoviral response. Short hairpin RNA (shRNA) lentiviral vectors targeting TBK1, STING, and cGAS were established in murine MS1 endothelial and RAW 264.7 macrophage cell lines. Knockdown of TBK1, STING, and cGAS results in a dramatic reduction in the activation of the primary antiviral response marker phosphorylated interferon (IFN) response factor 3 (IRF3) following exposure to adenovirus. Furthermore, activation of secondary type I IFN signaling targets (
ptyr
STAT1 and
ptyr
STAT2 [
ptyr
STAT1/2]) was also compromised. Consistent with compromised activation of primary and secondary response markers, transcriptional activation of IRF3-responsive genes (beta IFN [IFN-β], ISG15, ISG54) and secondary response transcripts were diminished in cells knocked down in cGAS, STING, or TBK1. These data establish cGAS as the dominant cytosolic DNA sensor responsible for detection of internalized adenovirus leading to induction of the type I interferon antiviral cascade.
Role of Palatine Tonsils as a Prion Entry Site in Classical and Atypical Experimental Sheep Scrapie
Authors: Cancedda Maria G. , Di Guardo Giovanni , Chiocchetti Roberto , Demontis Francesca , Marruchella Giuseppe , Sorteni Caterina , Maestrale Caterina , Lai Alfio , Ligios Ciriaco 
Published: November 2013

ABSTRACT

Atypical and classical scrapie-infected sheep brain tissue was monolaterally injected into the tonsils of lambs to investigate their role as a prion entry point. We first detected classical PrP
Sc
within the inoculated tonsil and in the ipsilateral retropharyngeal lymph node at 3 months postinoculation (p.i.). At 7 months p.i., PrP
Sc
colonized other lymphoid tissues bilaterally, including ileal Peyer's patches. The earliest PrP
Sc
deposition within the brain was ipsilaterally observed at 9 months p.i. in the substantia reticularis of the medulla oblongata. At 12 months p.i., PrP
Sc
deposition was present bilaterally in the nucleus parasympathicus nervi vagi, as well as in the intermediolateral cell column of the thoracolumbar spinal cord. No PrP
Sc
was detected in the lambs inoculated with atypical scrapie. These findings suggest that neuroinvasion may naturally occur from the tonsil after a widespread prion replication within the lymphoid tissues during classical scrapie only, thus mimicking the pathogenesis after oral ingestion.
The Human Alpha Defensin HD5 Neutralizes JC Polyomavirus Infection by Reducing Endoplasmic Reticulum Traffic and Stabilizing the Viral Capsid
Authors: Zins Stephen R. , Nelson Christian D. S. , Maginnis Melissa S. , Banerjee Rahul , O'Hara Bethany A. , Atwood Walter J. 
Published: November 2013

ABSTRACT
Progressive multifocal leukoencephalopathy (PML) is a fatal disease with limited treatment options, both clinically and in the research pipeline. Potential therapies would target and neutralize its etiologic agent, JC polyomavirus (JCPyV). The innate immune response to JCPyV infection has not been studied, and little is known about the initial host response to polyomavirus infection. This study examined the ability of a human alpha defensin, HD5, to neutralize JCPyV infection in human fetal glial cells. We show that HD5, by binding to the virion, blocks infection. The JCPyV-HD5 complexes bind to and enter host cells but are reduced in their ability to reach the endoplasmic reticulum (ER), where virions are normally uncoated. Furthermore, HD5 binding to the virion stabilizes the capsid and prevents genome release. Our results show that HD5 neutralizes JCPyV infection at an early postentry step in the viral life cycle by stabilizing the viral capsid and disrupting JCPyV trafficking. This study provides a naturally occurring platform for developing antivirals to treat PML and also expands on the known capabilities of human defensins.
Signaling Pathways in Murine Dendritic Cells That Regulate the Response to Vesicular Stomatitis Virus Vectors That Express Flagellin
Authors: Smedberg Jason R. , Westcott Marlena M. , Ahmed Maryam , Lyles Douglas S. 
Published: November 2013

ABSTRACT

Vesicular stomatitis virus (VSV) vectors that express heterologous antigens have shown promise as vaccines in preclinical studies. The efficacy of VSV-based vaccines can be improved by engineering vectors that enhance innate immune responses. We previously generated a VSV vaccine vector that incorporates two enhancing strategies: an M protein mutation (M51R) that prevents the virus from suppressing host antiviral responses and a gene encoding bacterial flagellin (M51R-F vector). The rationale was that intracellular expression of flagellin would activate innate immune pathways in addition to those activated by virus alone. This was tested with dendritic cells (DCs) from mice containing deletions in key signaling molecules. Infection of DC with either M51R or M51R-F vector induced the production of interleukin-12 (IL-12) and IL-6 and increased surface expression of T cell costimulatory molecules. These responses were dramatically reduced in DCs from IPS-1
−/−
mice. Infection with M51R-F vector also induced the production of IL-1β. In addition, in approximately half of the DCs, M51R-F vector induced pyroptosis, a proinflammatory-type of cell death. These responses to flagellin were ablated in DCs from NLRC4
−/−
mice but not Toll-like receptor 5-deficient (TLR5
−/−
) mice, indicating that they resulted from inflammasome activation. These results demonstrate that flagellin induces additional innate immune mechanisms over those induced by VSV alone.
A Lethal Disease Model for Hantavirus Pulmonary Syndrome in Immunosuppressed Syrian Hamsters Infected with Sin Nombre Virus
Authors: Brocato Rebecca L. , Hammerbeck Christopher D. , Bell Todd M. , Wells Jay B. , Queen Laurie A. , Hooper Jay W. 
Published: November 2013

ABSTRACTSin Nombre virus (SNV) is a rodent-borne hantavirus that causes hantavirus pulmonary syndrome (HPS) predominantly in North America. SNV infection of immunocompetent hamsters results in an asymptomatic infection; the only lethal disease model for a pathogenic hantavirus is Andes virus (ANDV) infection of Syrian hamsters. Efforts to create a lethal SNV disease model in hamsters by repeatedly passaging virus through the hamster have demonstrated increased dissemination of the virus but no signs of disease. In this study, we demonstrate that immunosuppression of hamsters through the administration of a combination of dexamethasone and cyclophosphamide, followed by infection with SNV, results in a vascular leak syndrome that accurately mimics both HPS disease in humans and ANDV infection of hamsters. Immunosuppressed hamsters infected with SNV have a mean number of days to death of 13 and display clinical signs associated with HPS, including pulmonary edema. Viral antigen was widely detectable throughout the pulmonary endothelium. Histologic analysis of lung sections showed marked inflammation and edema within the alveolar septa of SNV-infected hamsters, results which are similar to what is exhibited by hamsters infected with ANDV. Importantly, SNV-specific neutralizing polyclonal antibody administered 5 days after SNV infection conferred significant protection against disease. This experiment not only demonstrated that the disease was caused by SNV, it also demonstrated the utility of this animal model for testing candidate medical countermeasures. This is the first report of lethal disease caused by SNV in an adult small-animal model.
Promyelocytic Leukemia Protein Is a Cell-Intrinsic Factor Inhibiting Parvovirus DNA Replication
Authors: Mitchell Angela M. , Hirsch Matthew L. , Li Chengwen , Samulski R. Jude 
Published: November 2013

ABSTRACT
Tripartite motif proteins are important viral restriction factors and affect processes ranging from uncoating to transcription to immune signaling. Specifically, the promyelocytic leukemia protein (TRIM19; also called PML) is a viral restriction factor inhibiting processes from uncoating to transcription to cell survival. Here we investigated PML's effect on adeno-associated virus (AAV), a parvovirus used for gene delivery. Although dependovirus (AAV) and autonomous parvovirus (minute virus of mice) replication centers can colocalize with PML, PML's functional effect on parvoviruses is unknown. Using PML knockout mice, we determined that PML knockout enhances recombinant AAV2 (rAAV2) transduction at a range of vector doses in both male and female mice. In fact, male and female PML knockout mice exhibited up to 56-fold and 28-fold increases in transduction, respectively. PML inhibited several rAAV serotypes, suggesting a conserved mechanism, and organ specificity correlated with PML expression. Mechanistically, PML inhibited rAAV second-strand DNA synthesis, precluding inhibition of self-complementary rAAV, and did not affect the prior steps in transduction. Furthermore, we confirmed the effect of human PML on rAAV transduction through small interfering RNA (siRNA)-mediated knockdown in HuH7 cells and determined that the highest level of inhibition was due to effects of PML isoform II (PMLII). Overexpression of PMLII resulted in inhibition of second-strand synthesis, vector production, and genome replication. Moreover, wild-type AAV2 production and infectivity were also inhibited by PMLII, demonstrating a PML interaction with wild-type AAV. These data have important implications for AAV-mediated gene therapy. Additionally, PMLII inhibition of AAV second-strand synthesis and replication, which are processes necessary for all parvoviruses, suggests implications for replication of other parvoviruses.
Death Receptor-Mediated Apoptotic Signaling Is Activated in the Brain following Infection with West Nile Virus in the Absence of a Peripheral Immune Response
Authors: Clarke Penny , Leser J. Smith , Quick Eamon D. , Dionne Kalen R. , Beckham J. David , Tyler Kenneth L. 
Published: November 2013

ABSTRACT

Apoptosis is an important mechanism of West Nile virus (WNV) pathogenesis within the central nervous system (CNS). The signaling pathways that result in WNV-induced apoptotic neuronal death within the CNS have not been established. In this study, we identified death receptor (DR)-induced apoptosis as a pathway that may be important in WNV pathogenesis, based on the pattern of differential gene expression in WNV-infected, compared to uninfected, brains. Reverse transcription-PCR (RT-PCR) and Western blotting confirmed that genes involved in DR-induced apoptotic signaling are upregulated in the brain following WNV infection. Activity of the DR-associated initiator caspase, caspase 8, was also increased in the brains of WNV-infected mice and occurred in association with cleavage of Bid and activation of caspase 9. These results demonstrate that DR-induced apoptotic signaling is activated in the brain following WNV infection and suggest that the caspase 8-dependent cleavage of Bid promotes intrinsic apoptotic signaling within the brains of infected animals. Utilization of a novel
ex vivo
brain slice culture (BSC) model of WNV encephalitis revealed that inhibition of caspase 8 decreases virus-induced activation of caspase 3 and tissue injury. The BSC model allows us to examine WNV-induced pathogenesis in the absence of a peripheral immune response. Thus, our results indicate that WNV-induced neuronal injury in the brain is mediated by DR-induced apoptosis signaling and can occur in the absence of infiltrating immune cells. However, astrocytes and microglia were activated in WNV-infected BSC, suggesting that local immune responses influence WNV pathogenesis.
On Dermatomes, Meridians and Points: Results of a Quasiexperimental Study
Authors: Sánchez-Araujo Max , Luckert-Barela Ana J , Sánchez Nathalia , Torres Juan , Conde Jesus Eloy 
Published: November 2013

Background Traditional Chinese medicine (TCM) meridians and points run vertically, reflecting their function in the Zhang-Fu system (meridian pattern). However, the trunk's spinal nerves show a traverse orientation, or a ‘horizontal pattern’. Objective The aim of the present work was to evaluate, via a cognitive quasiexperiment, whether the clinical indications of the points on the trunk are associated with their meridian function or with their innervation and visceral–somatic connection. Methods The points in each dermatome of the trunk were considered crosswise, regardless of their meridians. The clinical indications for each point were differentiated into two mutually exclusive categories: (a) vertical distribution effect (VDE) or ‘meridian pattern’, when indications were quite different regarding the indications for the other points on the dermatome; and (b) transverse distribution effects (TDE) or ‘horizontal pattern’, represented by mainly local or segmental indications except for Shu-Mu points. After observing that the proportions between both categories often exceeded 60% in pilot samples, 60% was adopted as the reference value. Results A total of 22 dermatomes accommodated 148 points with 809 indications, of which 189 indications (23.4%) exhibited VDE features, whereas 620 (76.6%) exhibited TDE features. Conclusions A TDE/VDE ratio of 3 : 1 implies that the clinical indications for the points of any dermatome on the torso are similar, regardless of their meridians, and suggests that most of the indications for trunk points involve a ‘horizontal pattern’ due to their neurobiological nature. These findings may help in understanding acupuncture's neurobiology and clarify some confusing results of clinical research, for example, excluding sham acupuncture as an inert intervention for future clinical trials.
Challenges with cost-utility analyses of behavioural interventions among older adults at risk for dementia
Authors: Davis Jennifer C , Bryan Stirling , Marra Carlo A , Hsiung Ging-Yuek R , Liu-Ambrose Teresa 
Published: November 2013

BackgroundCognitive decline is one of the most prominent healthcare issues of the 21st century. Within the context of combating cognitive decline through behavioural interventions, physical activity is a promising approach. There is a dearth of health economic data in the area of behavioural interventions for dementia prevention. Yet, economic evaluations are essential for providing information to policy makers for resource allocation. It is essential we first address population and intervention-specific methodological challenges prior to building a larger evidence base. We use a cost-utility analysis conducted alongside the exercise for cognition and everyday living (EXCEL) study to illustrate methodological challenges specific to assessing the cost-effectiveness of behavioural interventions aimed at older adults at risk of cognitive decline.MethodsA cost-utility analysis conducted concurrently with a 6-month, three-arm randomised controlled trial (ie, the EXCEL study) was used as an example to identify and discuss methodological challenges.ResultsBoth the aerobic training and resistance training interventions were less costly than twice weekly balance and tone classes. In critically evaluating the economic evaluation of the EXCEL study we identified four category-specific challenges: (1) analysing costs; (2) assessing quality-adjusted life-years; (3) Incomplete data; and (4) ‘Intervention’ activities of the control group.ConclusionsResistance training and aerobic training resulted in healthcare cost saving and were equally effective to balance and tone classes after only 6 months of intervention. To ensure this population is treated fairly in terms of claims on resources, we first need to identify areas for methodological improvement.
The effect of a clinical pharmacist-led training programme on intravenous medication errors: a controlled before and after study
Authors: Nguyen Huong-Thao , Pham Hong-Tham , Vo Dang-Khoa , Nguyen Tuan-Dung , van den Heuvel Edwin R , Haaijer-Ruskamp Flora M , Taxis Katja 
Published: November 2013

BackgroundLittle is known about interventions to reduce intravenous medication administration errors in hospitals, especially in low- and middle-income countries.ObjectiveTo assess the effect of a clinical pharmacist-led training programme on clinically relevant errors during intravenous medication preparation and administration in a Vietnamese hospital.MethodsA controlled before and after study with baseline and follow-up measurements was conducted in an intensive care unit (ICU) and a post-surgical unit (PSU). The intervention comprised lectures, practical ward-based teaching sessions and protocols/guidelines, and was conducted by a clinical pharmacist and a nurse. Data on intravenous medication preparation and administration errors were collected by direct observation 12 h/day for seven consecutive days. Generalised estimating equations (GEE) were used to assess the effect of the intervention on the prevalence of clinically relevant erroneous doses, corrected for confounding factors.Results1204 intravenous doses were included, 516 during the baseline period (236 on ICU and 280 on PSU) and 688 during the follow-up period (407 on ICU and 281 on PSU). The prevalence of clinically relevant erroneous doses decreased significantly on the intervention ward (ICU) from 64.0% to 48.9% (p<0.001) but was unchanged on the control ward (PSU) (57.9% vs 64.1%; p=0.132). GEE analysis showed that doses on the intervention ward were 2.60 (1.27–5.31) times less likely to have clinically relevant errors (p=0.013).ConclusionsThe pharmacist-led training programme was effective, but the error rate remained relatively high. Further quality improvement strategies are needed, including changes to the working environment and promotion of a safety culture.
Temporal trends in the associations between age, sex and socioeconomic status after death from motor vehicle collisions in England and Wales: 1960–2009
Authors: Fogarty Andrew W , Liu Clarence 
Published: November 2013

ObjectiveTo determine the trend in the associations between socioeconomic status and gender with median age at death in England and Wales, from 1960 to 2009.MethodsAnnual cross-sectional studies of all registered deaths from a motor vehicle collision in England and Wales, 1960–2009.ResultsThere were 1647 deaths from a motor vehicle collision in 1960 and 964 deaths in 2009. The number of children aged 14 years or less who died in 1960 was 66 and this figure had reduced to 20 deaths by 2009. Individuals in non-manual occupations were consistently more likely to die above the median age of death than those in manual occupations during 1960–1963 (OR 1.66; 95% CI 1.50 to 1.84) and also during 1990–2000 (OR 1.54; 95% CI 1.44 to1.65). For 1960–1969, women had a higher risk of dying at above the annual median age of death (OR 1.72; 95% CI 1.62 to 1.82); for 2001–2009 the corresponding OR was 1.80 (95% CI 1.68 to 1.94).ConclusionsThere has been a 41% decrease in annual deaths after motor vehicle collisions in England and Wales over the past 50 years. The number of individuals over the age of 74 years dying in motor vehicle collisions has increased slightly, while the number of children's deaths decreased by 70% over the same time period despite driving becoming more common. Involvement in motor vehicle collisions may contribute to the sex and social class gradients in life expectancy observed in England and Wales.