bluuebunny/crossref_metadata_2025_split huggingface dataset:
Deletion of a 197-Amino-Acid Region in the N-Terminal Domain of Spike Protein Attenuates Porcine Epidemic Diarrhea Virus in Piglets
Authors: Hou Yixuan , Lin Chun-Ming , Yokoyama Masaru , Yount Boyd L. , Marthaler Douglas , Douglas Arianna L. , Ghimire Shristi , Qin Yibin , Baric Ralph S. , Saif Linda J. , Wang Qiuhong 
Published: May 2017

ABSTRACTWe previously isolated a porcine epidemic diarrhea virus (PEDV) strain, PC177, by blind serial passaging of the intestinal contents of a diarrheic piglet in Vero cell culture. Compared with the highly virulent U.S. PEDV strain PC21A, the tissue culture-adapted PC177 (TC-PC177) contains a 197-amino-acid (aa) deletion in the N-terminal domain of the spike (S) protein. We orally inoculated neonatal, conventional suckling piglets with TC-PC177 or PC21A to compare their pathogenicities. Within 7 days postinoculation, TC-PC177 caused mild diarrhea and lower fecal viral RNA shedding, with no mortality, whereas PC21A caused severe clinical signs and 55% mortality. To investigate whether infection with TC-PC177 can induce cross-protection against challenge with a highly virulent PEDV strain, all the surviving piglets were challenged with PC21A at 3 weeks postinoculation. Compared with 100% protection in piglets initially inoculated with PC21A, 88% and 100% TC-PC177- and mock-inoculated piglets had diarrhea following challenge, respectively, indicating incomplete cross-protection. To investigate whether this 197-aa deletion was the determinant for the attenuation of TC-PC177, we generated a mutant (icPC22A-S1Δ197) bearing the 197-aa deletion from an infectious cDNA clone of the highly virulent PEDV PC22A strain (infectious clone PC22A, icPC22A). In neonatal gnotobiotic pigs, the icPC22A-S1Δ197 virus caused mild to moderate diarrhea, lower titers of viral shedding, and no mortality, whereas the icPC22A virus caused severe diarrhea and 100% mortality. Our data indicate that deletion of this 197-aa fragment in the spike protein can attenuate a highly virulent PEDV, but the virus may lose important epitopes for inducing robust protective immunity.IMPORTANCEThe emerging, highly virulent PEDV strains have caused substantial economic losses worldwide. However, the virulence determinants are not established. In this study, we found that a 197-aa deletion in the N-terminal region of the S protein did not alter virus (TC-PC177) tissue tropism but reduced the virulence of the highly virulent PEDV strain PC22A in neonatal piglets. We also demonstrated that the primary infection with TC-PC177 failed to induce complete cross-protection against challenge by the highly virulent PEDV PC21A, suggesting that the 197-aa region may contain important epitopes for inducing protective immunity. Our results provide an insight into the role of this large deletion in virus propagation and pathogenicity. In addition, the reverse genetics platform of the PC22A strain was further optimized for the rescue of recombinant PEDV virusesin vitro. This breakthrough allows us to investigate other virulence determinants of PEDV strains and will provide knowledge leading to better control PEDV infections.
Cross-Linking of a CD4-Mimetic Miniprotein with HIV-1 Env gp140 Alters Kinetics and Specificities of Antibody Responses against HIV-1 Env in Macaques
Authors: Shen Xiaoying , Bogers Willy M. , Yates Nicole L. , Ferrari Guido , Dey Antu K. , Williams William T. , Jaeger Frederick H. , Wiehe Kevin , Sawant Sheetal , Alam S. Munir , LaBranche Celia C. , Montefiori David C. , Martin Loic , Srivastava Indresh , Heeney Jonathan , Barnett Susan W. , Tomaras Georgia D. 
Published: May 2017

ABSTRACTEvaluation of the epitope specificities, locations (systemic or mucosal), and effector functions of antibodies elicited by novel HIV-1 immunogens engineered to improve exposure of specific epitopes is critical for HIV-1 vaccine development. Utilizing an array of humoral assays, we evaluated the magnitudes, epitope specificities, avidities, and functions of systemic and mucosal immune responses elicited by a vaccine regimen containing Env cross-linked to a CD4-mimetic miniprotein (gp140-M64U1) in rhesus macaques. Cross-linking of gp140 Env to M64U1 resulted in earlier increases of both the magnitude and avidity of the IgG binding response than those with Env protein alone. Notably, IgG binding responses at an early time point correlated with antibody-dependent cellular cytotoxicity (ADCC) function at the peak immunity time point, which was higher for the cross-linked Env group than for the Env group. In addition, the cross-linked Env group developed higher IgG responses against a linear epitope in the gp120 C1 region of the HIV-1 envelope glycoprotein. These data demonstrate that structural modification of the HIV-1 envelope immunogen by cross-linking of gp140 with the CD4-mimetic M64U1 elicited an earlier increase of binding antibody responses and altered the specificity of the IgG responses, correlating with the rise of subsequent antibody-mediated antiviral functions.IMPORTANCEThe development of an efficacious HIV-1 vaccine remains a global priority to prevent new cases of HIV-1 infection. Of the six HIV-1 efficacy trials to date, only one has demonstrated partial efficacy, and immune correlate analysis of that trial revealed a role for binding antibodies and antibody Fc-mediated effector functions. New HIV-1 envelope immunogens are being engineered to selectively expose the most vulnerable and conserved sites on the HIV-1 envelope, with the goal of eliciting antiviral antibodies. Evaluation of the humoral responses elicited by these novel immunogen designs in nonhuman primates is critical for understanding how to improve upon immunogen design to inform further testing in human clinical trials. Our results demonstrate that structural modifications of Env that aim to mimic the CD4-bound conformation can result in earlier antibody elicitation, altered epitope specificity, and increased antiviral function postimmunization.
A Naturally Occurring Recombinant Enterovirus Expresses a Torovirus Deubiquitinase
Authors: Shang Pengcheng , Misra Saurav , Hause Ben , Fang Ying 
Published: May 2017

ABSTRACT

Enteroviruses (EVs) are implicated in a wide range of diseases in humans and animals. In this study, a novel enterovirus (enterovirus species G [EVG]) (EVG 08/NC_USA/2015) was isolated from a diagnostic sample from a neonatal pig diarrhea case and identified by using metagenomics and complete genome sequencing. The viral genome shares 75.4% nucleotide identity with a prototypic EVG strain (PEV9 UKG/410/73). Remarkably, a 582-nucleotide insertion, flanked by 3C
pro
cleavage sites at the 5′ and 3′ ends, was found in the 2C/3A junction region of the viral genome. This insertion encodes a predicted protease with 54 to 68% amino acid identity to torovirus (ToV) papain-like protease (PLP) (ToV-PLP). Structural homology modeling predicts that this protease adopts a fold and a catalytic site characteristic of minimal PLP catalytic domains. This structure is similar to those of core catalytic domains of the foot-and-mouth disease virus leader protease and coronavirus PLPs, which act as deubiquitinating and deISGylating (interferon [IFN]-stimulated gene 15 [ISG15]-removing) enzymes on host cell substrates. Importantly, the recombinant ToV-PLP protein derived from this novel enterovirus also showed strong deubiquitination and deISGylation activities and demonstrated the ability to suppress IFN-β expression. Using reverse genetics, we generated a ToV-PLP knockout recombinant virus. Compared to the wild-type virus, the ToV-PLP knockout mutant virus showed impaired growth and induced higher expression levels of innate immune genes in infected cells. These results suggest that ToV-PLP functions as an innate immune antagonist; enterovirus G may therefore gain fitness through the acquisition of ToV-PLP from a recombination event.

IMPORTANCE
Enteroviruses comprise a highly diversified group of viruses. Genetic recombination has been considered a driving force for viral evolution; however, recombination between viruses from two different orders is a rare event. In this study, we identified a special case of cross-order recombination between enterovirus G (order
Picornavirales
) and torovirus (order
Nidovirales
). This naturally occurring recombination event may have broad implications for other picornaviral and/or nidoviral species. Importantly, we demonstrated that the exogenous ToV-PLP gene that was inserted into the EVG genome encodes a deubiquitinase/deISGylase and potentially suppresses host cellular innate immune responses. Our results provide insights into how a gain of function through genetic recombination, in particular cross-order recombination, may improve the ability of a virus to evade host immunity.
Deletion of the Vaccinia Virus I2 Protein Interrupts Virion Morphogenesis, Leading to Retention of the Scaffold Protein and Mislocalization of Membrane-Associated Entry Proteins
Authors: Hyun Seong-In , Weisberg Andrea , Moss Bernard 
Published: May 2017

ABSTRACT
The I2L open reading frame of vaccinia virus (VACV) encodes a conserved 72-amino-acid protein with a putative C-terminal transmembrane domain. Previous studies with a tetracycline-inducible mutant demonstrated that I2-deficient virions are defective in cell entry. The purpose of the present study was to determine the step of replication or entry that is affected by loss of the I2 protein. Fluorescence microscopy experiments showed that I2 colocalized with a major membrane protein of immature and mature virions. We generated a cell line that constitutively expressed I2 and allowed construction of the VACV I2L deletion mutant vΔI2. As anticipated, vΔI2 was unable to replicate in cells that did not express I2. Unexpectedly, morphogenesis was interrupted at a stage after immature virion formation, resulting in the accumulation of dense spherical particles instead of brick-shaped mature virions with well-defined core structures. The abnormal particles retained the D13 scaffold protein of immature virions, were severely deficient in the transmembrane proteins that comprise the entry fusion complex (EFC), and had increased amounts of unprocessed membrane and core proteins. Total lysates of cells infected with vΔI2 also had diminished EFC proteins due to instability attributed to their hydrophobicity and failure to be inserted into viral membranes. A similar instability of EFC proteins had previously been found with unrelated mutants blocked earlier in morphogenesis that also accumulated viral membranes retaining the D13 scaffold. We concluded that I2 is required for virion morphogenesis, release of the D13 scaffold, and the association of EFC proteins with viral membranes.

IMPORTANCE
Poxviruses comprise a large family that infect vertebrates and invertebrates, cause disease in both in humans and in wild and domesticated animals, and are being engineered as vectors for vaccines and cancer therapy. In addition, investigations of poxviruses have provided insights into many aspects of cell biology. The I2 protein is conserved in all poxviruses that infect vertebrates, suggesting an important role. The present study revealed that this protein is essential for vaccinia virus morphogenesis and that its absence results in an accumulation of deformed virus particles retaining the scaffold protein and deficient in surface proteins needed for cell entry.
Roles of M1 and M2 Macrophages in Herpes Simplex Virus 1 Infectivity
Authors: Lee Dhong Hyun , Ghiasi Homayon 
Published: May 2017

ABSTRACTMacrophages are the predominant infiltrate in the corneas of mice that have been ocularly infected with herpes simplex virus 1 (HSV-1). However, very little is known about the relative roles of M1 (classically activated or polarized) and M2 (alternatively activated or polarized) macrophages in ocular HSV-1 infection. To better understand these relationships, we assessed the impact of directed M1 or M2 activation of RAW264.7 macrophages and peritoneal macrophages (PM) on subsequent HSV-1 infection. In both the RAW264.7 macrophage and PMin vitromodels, HSV-1 replication in M1 macrophages was markedly lower than in M2 macrophages and unstimulated controls. The M1 macrophages expressed significantly higher levels of 28 of the 32 tested cytokines and chemokines than M2 macrophages, with HSV-1 infection significantly increasing the levels of proinflammatory cytokines and chemokines in the M1 versus the M2 macrophages. To examine the effects of shifting the immune response toward either M1 or M2 macrophagesin vivo, wild-type mice were injected with gamma interferon (IFN-γ) DNA or colony-stimulating factor 1 (CSF-1) DNA prior to ocular infection with HSV-1. Virus replication in the eye, latency in trigeminal ganglia (TG), and markers of T cell exhaustion in the TG were determined. We found that injection of mice with IFN-γ DNA, which enhances the development of M1 macrophages, increased virus replication in the eye; increased latency; and also increased CD4, CD8, IFN-γ, and PD-1 transcripts in the TG of latently infected mice. Conversely, injection of mice with CSF-1 DNA, which enhances the development of M2 macrophages, was associated with reduced virus replication in the eye and reduced latency and reduced the levels of CD4, CD8, IFN-γ,and PD-1 transcripts in the TG. Collectively, these results suggest that M2 macrophages directly reduce the levels of HSV-1 latency and, thus, T-cell exhaustion in the TG of ocularly infected mice.IMPORTANCEOur findings demonstrate a novel approach to further reducing HSV-1 replication in the eye and latency in the TG by modulating immune components, specifically, by altering the phenotype of macrophages. We suggest that inclusion of CSF-1 as part of any vaccination regimen against HSV infection to coax responses of macrophages toward an M2, rather than an M1, response may further improve vaccine efficacy against ocular HSV-1 replication and latency.
Dok-1 and Dok-2 Are Required To Maintain Herpes Simplex Virus 1-Specific CD8+T Cells in a Murine Model of Ocular Infection
Authors: Lahmidi Soumia , Yousefi Mitra , Dridi Slimane , Duplay Pascale , Pearson Angela 
Published: May 2017

ABSTRACTDok-1 and Dok-2 negatively regulate responses downstream of several immune receptors in lymphoid and myeloid cells. Recent evidence showed that Dok proteins are essential in the formation of memory CD8+T cells to an exogenous epitope expressed by vaccinia virus; however, the importance of Dok-1 and Dok-2 in the control of viral infection is unknown. Here, we investigated the role of Dok proteins in modulating the immune response against herpes simplex virus 1 (HSV-1) in a mouse model of ocular infection. During acute infection, viral titers in the eye were similar in wild-type (WT) and Dok-1 and Dok-2 double-knockout (DKO) mice, and the percentages of infiltrating leukocytes were similar in DKO and WT corneas and trigeminal ganglia (TG). DKO mice exhibited a diminished CD8+T cell response to the immunodominant HSV-1 glycoprotein B (gB) epitope in the spleen and draining lymph nodes compared to WT mice during acute infection. Remarkably, gB-specific CD8+T cells almost completely disappeared in the spleens of DKO mice during latency, and the reduction of CD8+effector memory T (Tem) cells was more severe than that of CD8+central memory T (Tcm) cells. The percentage of gB-specific CD8+T cells in TG during latency was also dramatically reduced in DKO mice; however, they were phenotypically similar to those from WT mice. Inex vivoassays, reactivation was detected earlier in TG cultures from infected DKO versus WT mice. Thus, Dok-1 and Dok-2 promote survival of gB-specific CD8+T cells in TG latently infected with HSV-1.IMPORTANCEHSV-1 establishes lifelong latency in sensory neurons of trigeminal ganglia (TG). In humans, HSV-1 is able to sporadically reactivate from latently infected neurons and establish a lytic infection at a site to which the neurons project. Most herpetic disease in humans is due to reactivation of HSV-1 from latency rather than to primary acute infection. CD8+T cells are thought to play an important role in controlling recurrent infections. In this study, we examined the involvement of Dok-1 and Dok-2 signaling proteins in the control of HSV-1 infection. We provide evidence that Dok proteins are required to maintain a CD8+T cell response against HSV-1 during latency—especially CD8+Tem cells—and that they negatively affect HSV-1 reactivation from latency. Elucidating Dok-mediated mechanisms involved in the control of HSV-1 reactivation from latency might contribute to the development of therapeutic strategies to prevent recurrent HSV-1-induced pathology.
Congenital Human Cytomegalovirus Infection and the Enigma of Maternal Immunity
Authors: Britt William J. 
Published: May 2017

ABSTRACT
Human cytomegalovirus (HCMV) is the most common viral infection acquired by the developing human fetus and can result in damage to the developing central nervous system. Although vaccine development to modify this congenital infection is ongoing, the unique epidemiology of maternal HCMV infections appears discordant with strategies for vaccine development. Several characteristics of congenital HCMV infections suggest that the efficacy of vaccines designed to induce responses similar to those that follow natural infection will be limited.
Does treadmill running performance, heart rate and breathing rate response during maximal graded exercise improve after volitional respiratory muscle training?
Authors: Radhakrishnan K , Sharma VK , Subramanian SK 
Published: May 2017

AimMaximal physical exertion in sports usually causes fatigue in the exercising muscles, but not in the respiratory muscles due to triggering of the Respiratory muscle metabo-reflex, a sympathetic vasoconstrictor response leading to preferential increment in blood flow to respiratory muscles.1We planned to investigate whether a six week yogic pranayama based Volitional Respiratory Muscle Training (VRMT) can improve maximal Graded Exercise Treadmill Test (GXTT) performance in healthy adult recreational sportspersons.MethodsConsecutive, consenting healthy adult recreational sportspersons aged 20.56±2.49 years (n=30), volunteered to ‘baseline recording’ of resting heart rate (HR), blood pressure (BP), respiratory rate (RR), and Bruce ramp protocol maximal GXTT until volitional exhaustion providing total test time (TTT), derived VO2max, Metabolic Equivalent of Task (METs), HR and BP response during maximal GXTT and drop in recovery HR data. After six weeks of observation, they underwent ‘pre-intervention recording’ followed by supervised VRMT intervention for 6 weeks (30 minutes a day; 5 days a week) and then ‘post-intervention recording’. Repeated measures ANOVA with pairwise t statistical comparison was used to analyse the data.ResultsAfter supervised VRMT, we observed significant decrease in their resting supine RR (p<0.001), resting supine HR (p=0.001), HR after 5 minutes of assuming standing posture (p=0.003); significant increase in TTT (p<0.001), derived VO2max (p<0.001), METs (p<0.001) and drop in recovery HR (p=0.038); altered HR response and BP response during exercise.ConclusionsWe hypothesize that these changes are probably due to VRMT induced learnt behaviour to control the breathing pattern that improves breathing economy, improvement in respiratory muscle aerobic capacity, attenuation of respiratory muscle metabo-reflex, increase in cardiac stroke volume and autonomic resetting towards parasympatho-dominance. Yogic Pranayama based VRMT can be used in sports conditioning programme of athletes to further improve their maximal exercise performance, and as part of rehabilitation training during return from injury.
Selective impact of disease on short-term and long-term components of self-reported memory: a population-based HUNT study
Authors: Almkvist Ove , Bosnes Ole , Bosnes Ingunn , Stordal Eystein 
Published: May 2017

BackgroundSubjective memory is commonly considered to be a unidimensional measure. However, theories of performance-based memory suggest that subjective memory could be divided into more than one dimension.ObjectiveTo divide subjective memory into theoretically related components of memory and explore the relationship to disease.MethodsIn this study, various aspects of self-reported memory were studied with respect to demographics and diseases in the third wave of the HUNT epidemiological study in middle Norway. The study included all individuals 55 years of age or older, who responded to a nine-item questionnaire on subjective memory and questionnaires on health (n=18 633).ResultsA principle component analysis of the memory items resulted in two memory components; the criterion used was an eigenvalue above 1, which accounted for 54% of the total variance. The components were interpreted as long-term memory (LTM; the first component; 43% of the total variance) and short-term memory (STM; the second component; 11% of the total variance). Memory impairment was significantly related to all diseases (except Bechterew’s disease), most strongly to brain infarction, heart failure, diabetes, cancer, chronic obstructive pulmonary disease and whiplash. For most diseases, the STM component was more affected than the LTM component; however, in cancer, the opposite pattern was seen.ConclusionsSubjective memory impairment as measured in HUNT contained two components, which were differentially associated with diseases.
Four-year nationwide incidence of retinitis pigmentosa in South Korea: a population-based retrospective study from 2011 to 2014
Authors: Rim Tyler Hyungtaek , Park Hye Won , Kim Dong Wook , Chung Eun Jee 
Published: May 2017

ObjectiveTo determine the incidence of retinitis pigmentosa (RP) in South Korea.DesignNationwide, population-based retrospective study.SettingCensus population of South KoreaParticipantsThis study involved the entire population of South Korea (n=47 990 761). Patients confirmed as having RP by an ophthalmologist from 1 January 2011 to 31 December 2014 were included.Primary outcome measureThe average incidence of RP during the 4-year study period was estimated using population data from the 2010 Korean census.ResultsA total of 3144 (1567 men and 1577 women) patients confirmed as having RP were identified. The average incidence of RP was 1.64 cases/100 000 person-years (95% CI 1.58 to 1.70). The incidence of RP distribution skewed to the left across age groups, with one smaller peak observed in the 20–24-year-old age group (1.24 cases/100 000 person-years) and a larger peak observed in the 65–69-year-old age group (3.26 cases/100 000 person-years). The overall incidence was similar in men and women (1.64 cases/100 000 person-years (95% CI 1.56 to 1.73) for men; 1.63 cases/100 000 person-years (95% CI 1.55 to 1.72) for women).ConclusionsOur study’s estimates of the nationwide population-based incidence of RP in an Asian population will help advance the understanding of the disease onset and allow healthcare systems to plan accordingly.