bluuebunny/crossref_metadata_2025_split huggingface dataset:
Attenuation of Foot-and-Mouth Disease Virus by Engineered Viral Polymerase Fidelity
Authors: Rai Devendra K. , Diaz-San Segundo Fayna , Campagnola Grace , Keith Anna , Schafer Elizabeth A. , Kloc Anna , de los Santos Teresa , Peersen Olve , Rieder Elizabeth 
Published: May 2017

ABSTRACT

Foot-and-mouth disease virus (FMDV) RNA-dependent RNA polymerase (RdRp) (3D
pol
) catalyzes viral RNA synthesis. Its characteristic low fidelity and absence of proofreading activity allow FMDV to rapidly mutate and adapt to dynamic environments. In this study, we used the structure of FMDV 3D
pol
in combination with previously reported results from similar picornaviral polymerases to design point mutations that would alter replication fidelity. In particular, we targeted Trp237 within conserved polymerase motif A because of the low reversion potential inherent in the single UGG codon. Using biochemical and genetic tools, we show that the replacement of tryptophan 237 with phenylalanine imparts higher fidelity, but replacements with isoleucine and leucine resulted in lower-fidelity phenotypes. Viruses containing these W237 substitutions show
in vitro
growth kinetics and plaque morphologies similar to those of the wild-type (WT) A
24
Cruzeiro strain in BHK cells, and both high- and low-fidelity variants retained fitness during coinfection with the wild-type virus. The higher-fidelity W237F (W237F
HF
) mutant virus was more resistant to the mutagenic nucleoside analogs ribavirin and 5-fluorouracil than the WT virus, whereas the lower-fidelity W237I (W237I
LF
) and W237L
LF
mutant viruses exhibited lower ribavirin resistance. Interestingly, the variant viruses showed heterogeneous and slightly delayed growth kinetics in primary porcine kidney cells, and they were significantly attenuated in mouse infection experiments. These data demonstrate, for a single virus, that either increased or decreased RdRp fidelity attenuates virus growth in animals, which is a desirable feature for the development of safer and genetically more stable vaccine candidates.

IMPORTANCE
Foot-and-mouth disease (FMD) is the most devastating disease affecting livestock worldwide. Here, using structural and biochemical analyses, we have identified FMDV 3D
pol
mutations that affect polymerase fidelity. Recombinant FMDVs containing substitutions at 3D
pol
tryptophan residue 237 were genetically stable and displayed plaque phenotypes and growth kinetics similar to those of the wild-type virus in cell culture. We further demonstrate that viruses harboring either a W237F
HF
substitution or W237I
LF
and W237L
LF
mutations were highly attenuated in animals. Our study shows that obtaining 3D
pol
fidelity variants by protein engineering based on polymerase structure and function could be exploited for the development of attenuated FMDV vaccine candidates that are safer and more stable than strains obtained by selective pressure via mutagenic nucleotides or adaptation approaches.
Molluscum Contagiosum Virus MC159 Abrogates cIAP1-NEMO Interactions and Inhibits NEMO Polyubiquitination
Authors: Biswas Sunetra , Shisler Joanna L. 
Published: May 2017

ABSTRACT
Molluscum contagiosum virus (MCV) is a dermatotropic poxvirus that causes benign skin lesions. MCV lesions persist because of virally encoded immune evasion molecules that inhibit antiviral responses. The MCV MC159 protein suppresses NF-κB activation, a powerful antiviral response, via interactions with the NF-κB essential modulator (NEMO) subunit of the IκB kinase (IKK) complex. Binding of MC159 to NEMO does not disrupt the IKK complex, implying that MC159 prevents IKK activation via an as-yet-unidentified strategy. Here, we demonstrated that MC159 inhibited NEMO polyubiquitination, a posttranslational modification required for IKK and downstream NF-κB activation. Because MCV cannot be propagated in cell culture, MC159 was expressed independent of infection or during a surrogate vaccinia virus infection to identify how MC159 prevented polyubiquitination. Cellular inhibitor of apoptosis protein 1 (cIAP1) is a cellular E3 ligase that ubiquitinates NEMO. Mutational analyses revealed that MC159 and cIAP1 each bind to the same NEMO region, suggesting that MC159 may competitively inhibit cIAP1-NEMO interactions. Indeed, MC159 prevented cIAP1-NEMO interactions. MC159 also diminished cIAP1-mediated NEMO polyubiquitination and cIAP1-induced NF-κB activation. These data suggest that MC159 competitively binds to NEMO to prevent cIAP1-induced NEMO polyubiquitination. To our knowledge, this is the first report of a viral protein disrupting NEMO-cIAP1 interactions to strategically suppress IKK activation. All viruses must antagonize antiviral signaling events for survival. We hypothesize that MC159 inhibits NEMO polyubiquitination as a clever strategy to manipulate the host cell environment to the benefit of the virus.

IMPORTANCE
Molluscum contagiosum virus (MCV) is a human-specific poxvirus that causes persistent skin neoplasms. The persistence of MCV has been attributed to viral downregulation of host cell immune responses such as NF-κB activation. We show here that the MCV MC159 protein interacts with the NEMO subunit of the IKK complex to prevent NEMO interactions with the cIAP1 E3 ubiquitin ligase. This interaction correlates with a dampening of cIAP1 to polyubiquitinate NEMO and to activate NF-κB. This inhibition of cIAP1-NEMO interactions is a new viral strategy to minimize IKK activation and to control NEMO polyubiquitination. This research provides new insights into mechanisms that persistent viruses may use to cause long-term infection of host cells.
Natural Variation of Epstein-Barr Virus Genes, Proteins, and Primary MicroRNA
Authors: Correia Samantha , Palser Anne , Elgueta Karstegl Claudio , Middeldorp Jaap M. , Ramayanti Octavia , Cohen Jeffrey I. , Hildesheim Allan , Fellner Maria Dolores , Wiels Joelle , White Robert E. , Kellam Paul , Farrell Paul J. 
Published: May 2017

ABSTRACT
Viral gene sequences from an enlarged set of about 200 Epstein-Barr virus (EBV) strains, including many primary isolates, have been used to investigate variation in key viral genetic regions, particularly LMP1, Zp, gp350, EBNA1, and the BART microRNA (miRNA) cluster 2. Determination of type 1 and type 2 EBV in saliva samples from people from a wide range of geographic and ethnic backgrounds demonstrates a small percentage of healthy white Caucasian British people carrying predominantly type 2 EBV. Linkage of Zp and gp350 variants to type 2 EBV is likely to be due to their genes being adjacent to the EBNA3 locus, which is one of the major determinants of the type 1/type 2 distinction. A novel classification of EBNA1 DNA binding domains, named QCIGP, results from phylogeny analysis of their protein sequences but is not linked to the type 1/type 2 classification. The BART cluster 2 miRNA region is classified into three major variants through single-nucleotide polymorphisms (SNPs) in the primary miRNA outside the mature miRNA sequences. These SNPs can result in altered levels of expression of some miRNAs from the BART variant frequently present in Chinese and Indonesian nasopharyngeal carcinoma (NPC) samples. The EBV genetic variants identified here provide a basis for future, more directed analysis of association of specific EBV variations with EBV biology and EBV-associated diseases.

IMPORTANCE
Incidence of diseases associated with EBV varies greatly in different parts of the world. Thus, relationships between EBV genome sequence variation and health, disease, geography, and ethnicity of the host may be important for understanding the role of EBV in diseases and for development of an effective EBV vaccine. This paper provides the most comprehensive analysis so far of variation in specific EBV genes relevant to these diseases and proposed EBV vaccines. By focusing on variation in LMP1, Zp, gp350, EBNA1, and the BART miRNA cluster 2, new relationships with the known type 1/type 2 strains are demonstrated, and a novel classification of EBNA1 and the BART miRNAs is proposed.
A Point Mutation in the Rhesus Rotavirus VP4 Protein Generated through a Rotavirus Reverse Genetics System Attenuates Biliary Atresia in the Murine Model
Authors: Mohanty Sujit K. , Donnelly Bryan , Dupree Phylicia , Lobeck Inna , Mowery Sarah , Meller Jaroslaw , McNeal Monica , Tiao Greg 
Published: May 2017

ABSTRACT

Rotavirus infection is one of the most common causes of diarrheal illness in humans. In neonatal mice, rhesus rotavirus (RRV) can induce biliary atresia (BA), a disease resulting in inflammatory obstruction of the extrahepatic biliary tract and intrahepatic bile ducts. We previously showed that the amino acid arginine (R) within the sequence SRL (amino acids 445 to 447) in the RRV VP4 protein is required for viral binding and entry into biliary epithelial cells. To determine if this single amino acid (R) influences the pathogenicity of the virus, we generated a recombinant virus with a single amino acid mutation at this site through a reverse genetics system. We demonstrated that the RRV mutant (RRV
VP4-R446G
) produced less symptomatology and replicated to lower titers both
in vivo
and
in vitro
than those seen with wild-type RRV, with reduced binding in cholangiocytes. Our results demonstrate that a single amino acid change in the RRV VP4 gene influences cholangiocyte tropism and reduces pathogenicity in mice.

IMPORTANCE
Rotavirus is the leading cause of diarrhea in humans. Rhesus rotavirus (RRV) can also lead to biliary atresia (a neonatal human disease) in mice. We developed a reverse genetics system to create a mutant of RRV (RRV
VP4-R446G
) with a single amino acid change in the VP4 protein compared to that of wild-type RRV.
In vitro
, the mutant virus had reduced binding and infectivity in cholangiocytes.
In vivo
, it produced fewer symptoms and lower mortality in neonatal mice, resulting in an attenuated form of biliary atresia.
Deletion of the Vaccinia Virus B1 Kinase Reveals Essential Functions of This Enzyme Complemented Partly by the Homologous Cellular Kinase VRK2
Authors: Olson Annabel T. , Rico Amber B. , Wang Zhigang , Delhon Gustavo , Wiebe Matthew S. 
Published: May 2017

ABSTRACTThe vaccinia virus B1 kinase is highly conserved among poxviruses and is essential for the viral life cycle. B1 exhibits a remarkable degree of similarity to vaccinia virus-related kinases (VRKs), a family of cellular kinases, suggesting that the viral enzyme has evolved to mimic VRK activity. Indeed, B1 and VRKs have been demonstrated to target a shared substrate, the DNA binding protein BAF, elucidating a signaling pathway important for both mitosis and the antiviral response. In this study, we further characterize the role of B1 during vaccinia infection to gain novel insights into its regulation and integration with cellular signaling pathways. We begin by describing the construction and characterization of the first B1 deletion virus (vvΔB1) produced using a complementing cell line expressing the viral kinase. Examination of vvΔB1 revealed that B1 is critical for the production of infectious virions in various cell types and is sufficient for BAF phosphorylation. Interestingly, the severity of the defect in DNA replication following the loss of B1 varied between cell types, leading us to posit that cellular VRKs partly complement for the absence of B1 in some cell lines. Using cell lines devoid of either VRK1 or VRK2, we tested this hypothesis and discovered that VRK2 expression facilitates DNA replication and allows later stages of the viral life cycle to proceed in the absence of B1. Finally, we present evidence that the impact of VRK2 on vaccinia virus is largely independent of BAF phosphorylation. These data support a model in which B1 and VRK2 share additional substrates important for the replication of cytoplasmic poxviruses.IMPORTANCEViral mimicry of cellular signaling modulators provides clear evidence that the pathogen targets an important host pathway during infection. Poxviruses employ numerous viral homologs of cellular proteins, the study of which have yielded insights into signaling pathways used by both virus and cells alike. The vaccinia virus B1 protein is a homolog of cellular vaccinia virus-related kinases (VRKs) and is needed for viral DNA replication and likely other stages of the viral life cycle. However, much remains to be learned about how B1 and VRKs overlap functionally. This study utilizes new tools, including a B1 deletion virus and VRK knockout cells, to further characterize the functional links between the viral and cellular enzymes. As a result, we have discovered that B1 and VRK2 target a common set of substrates vital to productive infection of this large cytoplasmic DNA virus.
A Novel A(H7N2) Influenza Virus Isolated from a Veterinarian Caring for Cats in a New York City Animal Shelter Causes Mild Disease and Transmits Poorly in the Ferret Model
Authors: Belser Jessica A. , Pulit-Penaloza Joanna A. , Sun Xiangjie , Brock Nicole , Pappas Claudia , Creager Hannah M. , Zeng Hui , Tumpey Terrence M. , Maines Taronna R. 
Published: May 2017

ABSTRACT
In December 2016, a low-pathogenic avian influenza (LPAI) A(H7N2) virus was identified to be the causative source of an outbreak in a cat shelter in New York City, which subsequently spread to multiple shelters in the states of New York and Pennsylvania. One person with occupational exposure to infected cats became infected with the virus, representing the first LPAI H7N2 virus infection in a human in North America since 2003. Considering the close contact that frequently occurs between companion animals and humans, it was critical to assess the relative risk of this novel virus to public health. The virus isolated from the human case, A/New York/108/2016 (NY/108), caused mild and transient illness in ferrets and mice but did not transmit to naive cohoused ferrets following traditional or aerosol-based inoculation methods. The environmental persistence of NY/108 virus was generally comparable to that of other LPAI H7N2 viruses. However, NY/108 virus replicated in human bronchial epithelial cells with an increased efficiency compared with that of previously isolated H7N2 viruses. Furthermore, the novel H7N2 virus was found to utilize a relatively lower pH for hemagglutinin activation, similar to human influenza viruses. Our data suggest that the LPAI H7N2 virus requires further adaptation before representing a substantial threat to public health. However, the reemergence of an LPAI H7N2 virus in the northeastern United States underscores the need for continuous surveillance of emerging zoonotic influenza viruses inclusive of mammalian species, such as domestic felines, that are not commonly considered intermediate hosts for avian influenza viruses.

IMPORTANCE
Avian influenza viruses are capable of crossing the species barrier to infect mammals, an event of public health concern due to the potential acquisition of a pandemic phenotype. In December 2016, an H7N2 virus caused an outbreak in cats in multiple animal shelters in New York State. This was the first detection of this virus in the northeastern United States in over a decade and the first documented infection of a felid with an H7N2 virus. A veterinarian became infected following occupational exposure to H7N2 virus-infected cats, necessitating the evaluation of this virus for its capacity to cause disease in mammals. While the H7N2 virus was associated with mild illness in mice and ferrets and did not spread well between ferrets, it nonetheless possessed several markers of virulence for mammals. These data highlight the promiscuity of influenza viruses and the need for diligent surveillance across multiple species to quickly identify an emerging strain with pandemic potential.
Reducing musculoskeletal injury and concussion risk in schoolboy rugby players with a pre-activity movement control exercise programme: a cluster randomised controlled trial
Authors: Hislop Michael D , Stokes Keith A , Williams Sean , McKay Carly D , England Mike E , Kemp Simon P T , Trewartha Grant 
Published: May 2017

BackgroundInjury risk in youth rugby has received much attention, highlighting the importance of establishing evidence-based injury reduction strategies.AimTo determine the efficacy of a movement control exercise programme in reducing injuries in youth rugby players and to investigate the effect of programme dose on injury measures.MethodsIn a cluster-randomised controlled trial, 40 independent schools (118 teams, 3188 players aged 14–18 years) were allocated to receive either the intervention or a reference programme, both of which were to be delivered by school coaches. The intervention comprised balance training, whole-body resistance training, plyometric training, and controlled rehearsal of landing and cutting manoeuvres. Time-loss (>24 hours) injuries arising from school rugby matches were recorded by coaches and medical staff.Results441 time-loss match injuries (intervention, 233; control, 208) were reported across 15 938 match exposure-hours (intervention, 9083; control, 6855). Intention-to-treat results indicated unclear effects of trial arm on overall match injury incidence (rate ratio (RR)=0.85, 90% confidence limits 0.61 to 1.17), although clear reductions were evident in the intervention arm for concussion incidence (RR=0.71, 0.48 to 1.05). When trial arm comparisons were limited to teams who had completed three or more weekly programme sessions on average, clear reductions in overall match injury incidence (RR=0.28, 0.14 to 0.51) and concussion incidence (RR=0.41, 0.17 to 0.99) were noted in the intervention group.ConclusionA preventive movement control exercise programme can reduce match injury outcomes, including concussion, in schoolboy rugby players when compared with a standardised control exercise programme, although to realise the greatest effects players should complete the programme at least three times per week.
Pictorial prescribing reduces fentanyl drug administration errors: a simulated controlled study
Authors: Booth Stephen W , Gloag Maria , Kinna Sara , Bell Andrew , Wheble Joanna L C , Wheeler Daniel W 
Published: May 2017

ObjectivesTransmucosal fentanyl is used to treat transient exacerbations of cancer pain. Several immediate release products are available, presented as intranasal sprays, sublingual and buccal tablets, or lozenges. These are not interchangeable, creating potential for medication errors. We compared the incidence of medication errors in a simulated scenario using handwritten drug charts and charts labelled with preprinted self-adhesive stickers with full pictorial fentanyl prescriptions.Methods54 nurses were shown 5 handwritten drug charts and 5 with self-adhesive pictorial labels. Nurses indicated which preparation and dose they would administer from boxes of Instanyl, Abstral, Effentora and Actiq (Nycomed, ProStrakan, Cephalon and Teva, respectively). We measured the frequency of drug administration errors and asked them to rate the prescriptions for clarity on four-point Likert items.ResultsThe use of pictorial self-adhesive prescriptions significantly reduced errors in choice of preparation, from 20 with traditional handwritten charts to 6 with self-adhesive labels (OR 3.52, 95% CI 1.39 to 8.90, p=0.006), but the incidence of dose error was not significantly different (OR 1.47, 95% CI 0.80 to 2.70, p=0.281). Analysis of Likert items showed using pictorial printed labels significantly improved nurses’ understanding of choice of preparation, dose and maximum four hourly dose (p<0.0001, p=0.006 and p=0.028, respectively).ConclusionsThe use of pictorial prescribing appears to be a promising strategy that could reduce medication errors in choice of fentanyl preparations. There may be a wider use for pictorial prescribing where non-interchangeable preparations of the same drug exist.
Impact of perceived control on all-cause and cardiovascular disease mortality in three urban populations of Central and Eastern Europe: the HAPIEE study
Authors: Kozela Magdalena , Pająk Andrzej , Micek Agnieszka , Besala Agnieszka , Kubinova Ruzena , Malyutina Sofia , Tamosiunas Abdonas , Pikhart Hynek , Peasey Anne , Nikitin Yuri , Marmot Michael , Bobak Martin 
Published: May 2017

BackgroundInverse associations between perceived control and cardiovascular disease (CVD) have been reported in studies from Western Europe and the USA. To assess this relationship across different populations, we investigated the association between perceived control and all-cause and CVD mortality in three population-based cohorts of Eastern European countries.MethodsWe analysed data from a prospective cohort study in random population samples in Krakow (Poland), Novosibirsk (Russia) and six Czech towns. Baseline survey included structured questionnaire and objective examination in a clinic. Perceived control was assessed using an 11-item scale developed by the MacArthur Foundation Programme on Successful Midlife. Information on vital status was obtained from death registers. Effect of perceived control on mortality was assessed using Cox proportional hazards models.ResultsA total of 2377 deaths (1003 from CVD) occurred among 27 249 participants over a median 7-year follow-up. In the Czech and Polish cohorts, perceived control was inversely associated with mortality; the adjusted HRs for the lowest versus highest control quintiles were 1.71 (1.34 to 2.19) in men and 1.63 (1.14 to 2.35) in women for all-cause mortality and 2.31 (1.48 to 3.59) and 5.50 (2.14 to 14.13) for CVD deaths. There was no association between perceived control and mortality in Russia; the adjusted HRs for all-cause mortality were 1.03 (0.79 to 1.34) in men and 1.29 (0.82 to 2.02) in women.ConclusionsLow perceived control was associated with increased risk of all-cause and CVD mortality in Czech and Polish cohorts but not in Russia. It is possible that this inconsistency may partly reflect a different sociocultural understanding of the concept of control in Russia.
Finding maximum rank moment matrices by facial reduction on primal form and Douglas-Rachford iteration
Authors: Wang Fei , Reid Greg , Wolkowicz Henry 
Published: May 2017

Recent breakthroughs have been made in the use of semi-definite programming and its application to real polynomial solving. For example, the real radical of a zero dimensional ideal, can be determined by such approaches. Some progress has been made on the determination of the real radical in positive dimension by Ma, Wang and Zhi[5, 4]. Such work involves the determination of maximal rank semidefinite matrices. Existing methods are computationally expensive and have poorer accuracy on larger examples.
In previous work we showed that regularity in the form of the Slater constraint qualification (strict feasibility) fails for the moment matrix in the SDP feasibility problem[6]. We used facial reduction to obtain a smaller regularized problem for which strict feasibility holds. However we did not have a theoretical guarantee that our methods, based on facial reduction and Douglas-Rachford iteration ensured the satisfaction of the maximum rank condition.
Our work is motivated by the problems above. We discuss how to compute the moment matrix and its kernel using facial reduction techniques where the maximum rank property can be guaranteed by solving the dual problem. The facial reduction algorithms on the primal form is presented. We give examples that exhibit for the first time additional facial reductions beyond the first which are effective in practice with much better accuracy than SeDuMi(CVX).