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sumoylation promotes pml degradation during encephalomyocarditis virus infection.the promyelocytic leukemia (pml) protein is expressed in the diffuse nuclear fraction of the nucleoplasm and in matrix-associated structures, known as nuclear bodies (nbs). pml nb formation requires the covalent modification of pml to sumo. the noncovalent interactions of sumo with pml based on the identification of a sumo-interacting motif within pml seem to be required for further recruitment within pml nbs of sumoylated proteins. rna viruses whose replication takes place in the cytoplasm and ...201020826694
antiviral activity of bignoniaceae species occurring in the state of minas gerais (brazil): part 1.to evaluate the antiviral activity of bignoniaceae species occurring in the state of minas gerais, brazil.201020840554
polypyrimidine tract-binding protein stimulates the poliovirus ires by modulating eif4g binding.tethered hydroxyl-radical probing has been used to determine the orientation of binding of polypyrimidine tract-binding protein (ptb) to the poliovirus type 1 (mahoney) (pv-1(m)) internal ribosome entry site/segment (ires)-the question of which rna-binding domain (rbd) binds to which sites on the ires. the results show that under conditions in which ptb strongly stimulates ires activity, a single ptb is binding to the ires, a finding which was confirmed by mass spectrometry of ptb/ires complexes ...201020859255
nucleoporin phosphorylation triggered by the encephalomyocarditis virus leader protein is mediated by mitogen-activated protein kinases.cardioviruses disrupt nucleocytoplasmic transport through the activity of their leader (l) protein. we have shown that hyperphosphorylation of nuclear pore proteins (nucleoporins or nups), including nup62, nup153, and nup214, is central to this l protein function and requires one or more cytosolic kinases. in this study, potential cellular enzymes involved in encephalomyocarditis virus (emcv) l-directed nup phosphorylation were screened with a panel of specific, cell-permeating kinase inhibitors ...201020881039
viral induction of zac1b through tlr3- and irf3-dependent pathways.zinc finger protein regulator of apoptosis and cell cycle arrest (zac1) is a transcription factor able to induce apoptosis or cell cycle arrest through independent pathways. in spite of the important potential functions attributed to zac1, little is known of its physiological regulation and biological function. we discovered that variant zac1b was expressed in murine embryonic fibroblasts (mefs) treated with polyriboinosinic polyribocytidylic acid [poly(i:c)], a synthetic double-stranded rna. th ...201020947170
antiviral effects of a transgenic rna-dependent rna polymerase.transgenic expression of the rna-dependent rna polymerase 3d(pol) inhibited infection of theiler's murine encephalitis virus (tmev), a picornavirus from which it was derived. here, we infected 3d(pol) transgenic mice with another picornavirus, as well as an alphaherpesvirus and a rhabdovirus. 3d(pol) transgenic fvb mice had significantly lower viral loads and survived longer after infection with all three types of viruses than nontransgenic fvb mice. viral inhibition among three different types ...201020962089
further observations on the virus of encephalomyocarditis. 194621064731
anti-infective activities of pelargonium sidoides (eps® 7630): effects of induced no production on leishmania major in infected macrophages and antiviral effects as assessed in a fibroblast-virus protection assay.eps® 7630 is an aqueous-ethanolic extract of the roots of pelargonium sidoides, employed in the treatment of upper respiratory tract infections. its anti-infective activity is supposed to be associated with the activation of the nonspecific immune system. using leishmania major gfp-infected murine bmmφ, the no production of eps® 7630-activated macrophages was correlated with the reduction of the gfp signal measured at single cell levels using flow cytometry. the anti-infectious effect of eps® 76 ...201021104606
the burden of obesity on infectious disease.the world is now experiencing an epidemic of obesity. although the effects of obesity on the development of metabolic and cardiovascular problems are well studied, much less is known about the impact of obesity on immune function and infectious disease. studies in obese humans and with obese animal models have repeatedly demonstrated impaired immune function, including decreased cytokine production, decreased response to antigen/mitogen stimulation, reduced macrophage and dendritic cell function ...201021127339
alkylated porphyrins have broad antiviral activity against hepadnaviruses, flaviviruses, filoviruses, and arenaviruses.we screened ∼2,200 compounds known to be safe in people for the ability to reduce the amount of virion-associated hepatitis b virus (hbv) dna in the culture medium of producer cells. these efforts led to the discovery of an alkylated porphyrin, chlorophyllide, as the compound that achieved the greatest reduction in signal. here we report that chlorophyllide directly and quantitatively disrupted hbv virions at micromolar concentrations, resulting in the loss of all detectable virion dna, without ...201021135183
exendin-4, a glucagon-like peptide-1 receptor agonist, suppresses pancreatic β-cell destruction induced by encephalomyocarditis virus.viral infection is one of the important factors for the pathogenesis of type 1 diabetes. particularly, in fulminant type 1 diabetes, rapid β-cell destruction is suggested to be triggered by viral infection. recently, glucagon-like peptide 1 (glp-1) receptor agonists have been reported to have direct beneficial effects on β-cells, such as anti-apoptotic effect, increasing β-cell mass, and improvement of β-cell function. however, their effects on β-cell destruction induced by viral infections have ...201021144822
mutational analysis of the emcv 2a protein identifies a nuclear localization signal and an eif4e binding site.cardioviruses have a unique 2a protein (143 aa). during genome translation, the encephalomyocarditis virus (emcv) 2a is released through a ribosome skipping event mitigated through c-terminal 2a sequences and by subsequent n-terminal reaction with viral 3c(pro). although viral replication is cytoplasmic, mature 2a accumulates in nucleoli shortly after infection. some protein also transiently associates with cytoplasmic 40s ribosomal subunits, an activity contributing to inhibition of cellular ca ...201021145089
a novel role for ifn-stimulated gene factor 3ii in ifn-γ signaling and induction of antiviral activity in human cells.type i (e.g., ifn-α, ifn-β) and type ii ifns (ifn-γ) have antiviral, antiproliferative, and immunomodulatory properties. both types of ifn signal through the jak/stat pathway to elicit antiviral activity, yet ifn-γ is thought to do so only through stat1 homodimers, whereas type i ifns activate both stat1- and stat2-containing complexes such as ifn-stimulated gene factor 3. in this study, we show that ifn-stimulated gene factor 3 containing unphosphorylated stat2 (isgf3(ii)) also plays a role in ...201021178011
pandemic 2009 h1n1 influenza virus is resistant to the antiviral activity of several interferon alpha subtypes.interferons (ifns) are critically important in the control of influenza a virus infections. to better understand the pathogenic characteristics of the pandemic 2009 h1n1 influenza virus (ph1n1) from an innate immunity viewpoint, we investigated whether in vitro ph1n1 is sensitive to the antiviral activity of ifn beta, leukocyte ifn alpha, and several ifn alpha subtypes in a human lung adenocarcinoma epithelial cell line under single-growth cycle conditions. the results showed that 50% inhibitory ...201121235413
recognition of cytoplasmic rna results in cathepsin-dependent inflammasome activation and apoptosis in human macrophages.dsrna is an important pathogen-associated molecular pattern that is primarily recognized by cytosolic pattern-recognition receptors of the innate-immune system during virus infection. this recognition results in the activation of inflammasome-associated caspase-1 and apoptosis of infected cells. in this study, we used high-throughput proteomics to identify secretome, the global pattern of secreted proteins, in human primary macrophages that had been activated through the cytoplasmic dsrna-recogn ...201121257972
mitochondrial membrane potential is required for mavs-mediated antiviral signaling.mitochondria, dynamic organelles that undergo cycles of fusion and fission, are the powerhouses of eukaryotic cells and are also involved in cellular innate antiviral immunity in mammals. mitochondrial antiviral immunity depends on activation of the cytoplasmic retinoic acid-inducible gene i (rig-i)-like receptor (rlr) signaling pathway and the participation of a mitochondrial outer membrane adaptor protein called mavs (mitochondrial antiviral signaling). we found that cells that lack the abilit ...201121285412
the nlrp3 inflammasome detects encephalomyocarditis virus and vesicular stomatitis virus infection.inflammasomes are cytosolic protein complexes that regulate caspase-1 activation and the secretion of interleukin-1ß (il-1ß) and il-18. several different inflammasome complexes have been identified, but the nlrp3 inflammasome is particularly notable because of its central role in diseases of inflammation. recent work has demonstrated an essential role for the nlrp3 inflammasome in host defense against influenza virus. we show here that two other rna viruses, encephalomyocarditis virus (emcv) and ...201121289120
common conformational changes induced in type 2 picornavirus iress by cognate trans-acting factors.type 2 internal ribosomal entry sites (iress) of encephalomyocarditis virus (emcv), foot-and-mouth disease virus (fmdv) and other picornaviruses comprise five major domains h-l. initiation of translation on these iress begins with specific binding of the central domain of initiation factor, eif4g to the j-k domains, which is stimulated by eif4a. eif4g/eif4a then restructure the region of ribosomal attachment on the ires and promote recruitment of ribosomal 43s pre-initiation complexes. in additi ...201121306989
the irf-3/bax-mediated apoptotic pathway, activated by viral cytoplasmic rna and dna, inhibits virus replication.induction of apoptosis in cells infected by sendai virus (sev), which triggers the cytosolic rig-i pathway, requires the presence of interferon regulatory factor 3 (irf-3). independent of irf-3's transcriptional role, a novel irf-3 activation pathway causes its interaction with the proapoptotic protein bax and its mitochondrial translocation to induce apoptosis. here we report that two other rna viruses, vesicular stomatitis virus (vsv) and encephalomyocarditis virus (emcv), may also activate th ...201121307205
encephalomyocarditis virus mortality in semi-wild bonobos (pan panicus).fatal myocarditis from encephalomyocarditis virus (emcv) infection has previously been identified in sporadic and epidemic forms in many species of captive non-human primates probably including one bonobo (pan paniscus).201121323673
in vitro and in vivo comparison of viral and cellular internal ribosome entry sites for bicistronic vector expression.bicistronic vectors are essential to achieve efficient expression of multiple genes in gene therapy protocols and biomedical applications. internal ribosome entry site (ires) elements have been utilized to initiate expression of an additional protein from a bicistronic vector. the ires element commonly used in current bicistronic vectors originates from the encephalomyocarditis virus (emcv). as ires-mediated translation is dependent on availability of ires trans-acting factors, which vary betwee ...201121368899
prevalence and risk factors for encephalomyocarditis virus infection in peru.although encephalomyocarditis virus (emcv) infection has been commonly documented among domestic animals, less is known about emcv transmission among humans. recently, we described the isolation of emcv from two febrile patients in peru. to further investigate emcv transmission in peru, we screened febrile patients reporting to health clinics in peru for serological evidence of recent emcv infection. we also conducted a serological survey for emcv-neutralizing antibodies in the city of iquitos, ...201121395427
efficient co-expression of bicistronic proteins in mesenchymal stem cells by development and optimization of a multifunctional plasmid.local synthesis of interferon within b16 tumors mediates anti-tumor effects. based on reports that stem cells are recruited to tumors, and because systemic administration of interferon causes dose-limiting undesirable side effects, we wanted to improve the anti-tumor effects of interferon while simultaneously minimizing its systemic side effects by employing mesenchymal stem cells (mscs) as tumor-localized ectopic producers of interferon. many vectors exist to fulfill this purpose, but their tra ...201121401924
rna sensor-induced type i ifn prevents diabetes caused by a ß cell-tropic virus in mice.viral infections have been linked to the onset of type i diabetes (t1d), with viruses postulated to induce disease directly by causing ß cell injury and subsequent release of autoantigens and indirectly via the host type i interferon (ifn-i) response triggered by the virus. consistent with this, resistance to t1d is associated with polymorphisms that impair the function of melanoma differentiation associated gene-5 (mda5), a sensor of viral rna that elicits ifn-i responses. in animal models, tri ...201121403398
possible role of phosphoinositide-3-kinase in mx1 protein translation and antiviral activity of interferon-omega-stimulated hela cells.interferon ? (ifn-?), a cytokine released during innate immune activation, is well known for promoting direct antiviral responses; however, the possible signal pathways that are initiated by ifn-? binding to the type i ifn receptors have not been fully studied. here, we provide evidence that activation of phosphoinositide-3-kinase/protein kinase b (pi3k/akt) signaling plays a pivotal role in the generation of ifn-?-mediated biological responses. we found that ly294002 (pi3k inhibitor)-attenuated ...201121430412
autophagy promotes the replication of encephalomyocarditis virus in host cells.a growing number of studies have demonstrated that autophagy has a diverse role in the infection process of different pathogens. however, to date, it is unknown whether autophagy is activated in encephalomyocarditis virus (emcv)-infected host cells, and if so, what its role is in this process. in the present study, we first demonstrated that emcv infection significantly increases the number of double- and single-membrane vesicles in the cytoplasm of host cells. it was then confirmed that these o ...201121460631
immunogenicity and safety of virus-like particle of the porcine encephalomyocarditis virus in pig.abstract:201121492483
microbiological safety of porcine islets: comparison with source pig.pig islet donors intended for clinical xenotransplantation for the treatment of diabetes must meet stringent conditions. among others, viruses with the potential to cross the species barrier should be excluded from the herd: this list includes encephalomyocarditis virus (emcv), hepatitis e virus (hev), porcine cytomegalovirus (pcmv) and porcine γ-lymphotropic herpesvirus (plhv). as an islet product is isolated from the pancreas and then subjected to culture before implantation, the question is r ...201121496116
activation of picornaviral iress by ptb shows differential dependence on each ptb rna-binding domain.polypyrimidine tract binding protein (ptb) is an rna-binding protein with four rna-binding domains (rbds). it is a major regulator of alternative splicing and also stimulates translation initiation at picornavirus iress (internal ribosome entry sites). the sites of interaction of each rbd with two picornaviral iress have previously been mapped. to establish which rbd-ires interactions are essential for ires activation, point mutations were introduced into the rna-binding surface of each rbd. thr ...201121518806
adaptation of saffold virus-2 for high-titer growth in mammalian cells.saffold viruses (safv) are a recently discovered group of human cardioviruses closely related to the theiler's murine encephalomyelitis viruses (tmev). unlike tmev and encephalomyocarditis virus, each of which is monotypic, safv are genetically diverse and include at least eight genotypes. to date, only safv-3 has been grown efficiently in mammalian cells in vitro. here, we report the successful adaptation of safv-2 for efficient growth in hela cells after 13 passages in the interferon α/β-defic ...201121543476
the protease-sensitive loop of the vesicular stomatitis virus matrix protein is involved in virus assembly and protein translation.to study the contribution of the protease-sensitive loop of the vsv m protein in virus assembly we recovered recombinant vsv (rvsv) with mutations in this region and examined virus replication. mutations in the highly conserved lxd motif (aa 123-125) resulted in reduced virion budding, reduced virus titers and enhanced m protein exchange with m-ribonucleocapsid complexes (m-rnps), suggesting that the mutant m proteins were less tightly associated with rnp skeletons. in addition, viral protein sy ...201121596416
recombinant protein hydrazides: application to site-specific protein pegylation.here, we describe a novel method for the site-specific c-terminal pegylation of recombinant proteins. this general approach exploits chemical cleavage of precursor intein-fusion proteins with hydrazine to directly produce recombinant protein hydrazides. this unique functionality within the protein sequence then facilitates site-specific c-terminal modification by hydrazone-forming ligation reactions. this approach was used to generate folded, site-specifically c-terminal pegylated ifnalpha2b and ...201121618993
inhibition of rnase l and pkr by sunitinib impairs antiviral innate immunity.rnase l and pkr are effectors of the interferon antiviral response that share homology in their pseudokinase and protein kinase domains, respectively. sunitinib is an orally available, atp competitive inhibitor of vegf-r and pdgf-r used clinically to suppress angiogenesis and tumor growth. sunitinib also impacts ire1, an endoplasmic reticulum protein involved in the unfolded protein response that is closely-related to rnase l. here we report that sunitinib is a potent inhibitor of both rnase l a ...201121636578
epizootic myocarditis associated with encephalomyocarditis virus in a group of rhesus macaques (macaca mulatta).six cases of fatal myocarditis associated with encephalomyocarditis virus occurred over a 14-month period in a group of outdoor-housed juvenile rhesus macaques. all animals were younger than 3 years of age and died or were euthanized following acute onset of dyspnea or pulmonary effusion (3 of 6) or were found dead without premonitory signs (3 of 6). gross findings included pulmonary congestion (6 of 6), variable degrees of pleural effusion (4 of 6), multifocal pale tan foci throughout the myoca ...201121653204
immunogenicity of an interferon-beta1a product.in order to determine whether blastoferon®, a biosimilar interferon (ifn)- beta 1a formulation, shares epitopes with other known ifn-beta products, a series of neutralization bioassays were performed with a set of well-characterized anti-ifn- beta monoclonal antibodies and human sera (world health organization reference reagents). the bioassay was the interferon-induced inhibition of virus cytopathic effect on human cells in culture (emc virus and a-549 cells). computer-calculated results were r ...201121658324
kshv virf-2 inhibits type 1 interferon signalling by targeting isgf-3.kaposi's sarcoma-associated herpesvirus (kshv) encodes four viral interferon regulatory factors (virf-1-4). we investigated the mechanism and consequences of virf-2-mediated inhibition of interferon-response element (isre) signalling following type i interferon (ifn) induction. western blot and electrophoretic mobility shift assays identified the interferon stimulated gene factor-3 (isgf-3) components stat-1 and irf-9 as the proximal targets of virf-2 activity. the biological significance of vir ...201121697347
alteration of encephalomyocarditis virus pathogenicity due to a mutation at position 100 of vp1.encephalomyocarditis virus (emcv) infection leads to many diseases including encephalitis, myocarditis and diabetes in its natural host, the mouse. in this study, we generated four cdna clones with a point mutation at position 100 of vp1. the amino acids isoleucine, alanine, serine and proline were substituted with threonine in the four different clones of emcv strain bjc3 by site-specific mutagenesis, and viable viruses were rescued. although all mutants and wild-type viruses display different ...201121706414
compare the differences of synonymous codon usage between the two species within cardiovirus.abstract:201121708006
knockdown of usp18 increases alpha 2a interferon signaling and induction of interferon-stimulating genes but does not increase antiviral activity in huh7 cells.the current standard of care for hepatitis c virus (hcv) patients is cotreatment with human alpha interferon (ifn-α) and ribavirin. the host factor usp18 functions to regulate the interferon signaling pathway by acting as an off-switch. in order to understand whether the inhibition of usp18 represents a valid target for the enhancement of interferon treatment for chronic viral diseases, we have used a wide range of rna interference (rnai) reagents to suppress usp18 gene expression in huh7 cell l ...201121709085
the effect of fty720 in the theiler's virus model of multiple sclerosis.fty720 (fingolimod) has demonstrated efficacy in multiple sclerosis (ms). we evaluated the effects of fty720 on progressive disability, viral load, and antibody responses in mice infected with theiler's murine encephalomyocarditis virus (tmev). fty720 and phosphorylated fty720 (fty720-p) were detected in the brain after intraperitoneal injection of the drug. bioactivity of fty720 was confirmed by reduced numbers of mononuclear cells in the spleen and blood after treatment. no significant differe ...201121726878
Involvement of Noxa in mediating cellular ER stress responses to lytic virus infection.Noxa is a Bcl-2 homology domain-containing pro-apoptotic mitochondrial protein. Noxa mRNA and protein expression are upregulated by dsRNA or virus, and ectopic Noxa expression enhances cellular sensitivity to virus or dsRNA-induced apoptosis. Here we demonstrate that Noxa null baby mouse kidney (BMK) cells are deficient in normal cytopathic response to lytic viruses, and that reconstitution of the knockout cells with wild-type Noxa restored normal cytopathic responses. Noxa regulation by virus m ...201121742363
saffold cardioviruses in children with diarrhea, thailand.to the editor: cardioviruses currently consist of at least 3 viruses: theiler murine encephalomyocarditis virus, encephalomyocarditis virus, and saffold virus (safv) (1-4). saffold cardiovirus in the family picornaviridae was isolated and identified from fecal specimens of a child with fever of unknown origin in the united states (3).201121749802
the leader protein of cardioviruses inhibits stress granule assembly.stress granules (sg) are cytoplasmic aggregates of stalled translation pre-initiation complexes forming in cells exposed to various environmental stresses. here, we show that stress granules assemble in cells infected with theiler's murine encephalomyelitis virus (tmev) mutants carrying alterations in the leader (l) protein, but not in cells infected with wild-type tmev. stress granules also formed in stat1-deficient cells, suggesting that sg formation was not a consequence of increased type-i i ...201121752908
translation of viral mrna without active eif2: the case of picornaviruses.previous work by several laboratories has established that translation of picornavirus rna requires active eif2a for translation in cell free systems or after transfection in culture cells. strikingly, we have found that encephalomyocarditis virus protein synthesis at late infection times is resistant to inhibitors that induce the phosphorylation of eif2a whereas translation of encephalomyocarditis virus early during infection is blocked upon inactivation of eif2a by phosphorylation induced by a ...201121779397
studies of inactivation of encephalomyocarditis virus, m13 bacteriophage, and salmonella typhimurium by using a visible femtosecond laser: insight into the possible inactivation mechanisms.we report experimental results on the inactivation of encephalomyocarditis virus, m13 bacteriophage, and salmonella typhimurium by a visible femtosecond laser. our results suggest that inactivation of virus and bacterium by a visible femtosecond laser involves completely different mechanisms. inactivation of viruses by a visible femtosecond laser involves the breaking of hydrogen/hydrophobic bonds or the separation of the weak protein links in the protein shell of a viral particle. in contrast, ...201121806295
novel chikungunya vaccine candidate with an ires-based attenuation and host range alteration mechanism.chikungunya virus (chikv) is a reemerging mosquito-borne pathogen that has recently caused devastating urban epidemics of severe and sometimes chronic arthralgia. as with most other mosquito-borne viral diseases, control relies on reducing mosquito populations and their contact with people, which has been ineffective in most locations. therefore, vaccines remain the best strategy to prevent most vector-borne diseases. ideally, vaccines for diseases of resource-limited countries should combine lo ...201121829348
encephalomyocarditis virus 2a protein is required for viral pathogenesis and inhibition of apoptosis.the encephalomyocarditis virus (emcv), a picornaviridae virus, has a wide host spectrum and can cause various diseases. emcv virulence factors, however, are as yet ill defined. here, we demonstrate that the emcv 2a protein is essential for the pathogenesis of emcv. infection of mice with the b279/95 strain of emcv resulted in acute fatal disease, while the clone c9, derived by serial in vitro passage of the b279/95 strain, was avirulent. c9 harbored a large deletion in the gene encoding the 2a p ...201121849462
antiviral activity of distictella elongata (vahl) urb. (bignoniaceae), a potentially useful source of anti-dengue drugs from the state of minas gerais, brazil.to investigate the in vitro antiviral activity of distictella elongata (vahl) urb. ethanol extracts from leaves (lee), fruits (fee), stems and their main components.201121895729
Unusual loop-sequence flexibility of the proximal RNA replication element in EMCV.Picornaviruses contain stable RNA structures at the 5' and 3' ends of the RNA genome, OriL and OriR involved in viral RNA replication. The OriL RNA element found at the 5' end of the enterovirus genome folds into a cloverleaf-like configuration. In vivo SELEX experiments revealed that functioning of the poliovirus cloverleaf depends on a specific structure in this RNA element. Little is known about the OriL of cardioviruses. Here, we investigated structural aspects and requirements of the apical ...201121935472
Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube.Virus particles are used in vaccination, drug delivery, and material sciences. Here we devised a system where the RNA virus encephalomyocarditis virus (EMCV) is synthesized from DNA templates in vitro. When a plasmid or a PCR product harboring the full-length cDNA of EMCV in the T7 promoter/terminator unit was incubated in a HeLa cell extract supplemented with T7 RNA polymerase, EMCV was produced within 4 h at an efficiency of over 10-fold compared with the system programmed with the EMCV RNA. T ...201221952914
conditional transgenic expression of tir-domain-containing adaptor-inducing interferon-β (trif) in the adult mouse heart is protective in acute viral myocarditis.tir-domain-containing adaptor-inducing interferon-β (trif) plays a major role in toll-like receptor 3 (tlr3) mediated signaling. mice deficient in tlr3 and trif have been shown to be highly susceptible to enterovirus-induced myocardial injury. these mice have decreased production of antiviral cytokines and increased viral replication in the heart. therefore, we hypothesized that conditional overexpression of trif would change cardiac myocyte susceptibility to virus infection by augmenting the an ...201121956162
encephalomyocarditis virus may use different pathways to initiate infection of primary human cardiomyocytes.encephalomyocarditis virus (emcv) can infect a wide range of vertebrate species including swine and non-human primates, but few data are available for humans. we therefore wanted to gain further insight into the mechanisms involved in emcv infection of human cells. for this purpose, we analyzed the permissiveness of primary human cardiomyocytes towards two strains of emcv; a pig myocardial strain (b279/95) and a rat strain (1086c). in this study, we show that both strains productively infect pri ...201121989795
promyelocytic leukemia isoform iv confers resistance to encephalomyocarditis virus via the sequestration of 3d polymerase in nuclear bodies.promyelocytic leukemia (pml) protein is the organizer of nuclear matrix-associated nuclear bodies (nbs), and its conjugation to the small ubiquitin-like modifier (sumo) is required for the formation of these structures. several alternatively spliced pml transcripts from a single pml gene lead to the production of seven pml isoforms (pml isoform i [pmli] to vii [pmlvii]), which all share a n-terminal region that includes the rbcc (ring, b boxes, and a α-helical coiled-coil) motif but differ in th ...201121994459
ccr5 regulates inflammatory gene expression in response to encephalomyocarditis virus infection.encephalomyocarditis virus (emcv) is capable of stimulating inflammatory gene expression by macrophages as a result of interactions between emcv capsid proteins and cell surface receptors. in this study, biochemical and genetic approaches identified a role for ccr5, a chemokine receptor, in transducing the signals of emcv infection that result in the expression of inflammatory genes in macrophages. antibody neutralization and gene knockout strategies were used to show that the presence of ccr5 i ...201122001348
Ribosomal frameshifting into an overlapping gene in the 2B-encoding region of the cardiovirus genome.The genus Cardiovirus (family Picornaviridae) currently comprises the species Encephalomyocarditis virus (EMCV) and Theilovirus. Cardioviruses have a positive-sense, single-stranded RNA genome that encodes a large polyprotein (L-1ABCD-2ABC-3ABCD) that is cleaved to produce approximately 12 mature proteins. We report on a conserved ORF that overlaps the 2B-encoding sequence of EMCV in the +2 reading frame. The ORF is translated as a 128-129 amino acid transframe fusion (2B*) with the N-terminal 1 ...201122025686
inhibition of encephalomyocarditis virus replication by shrna targeting 1d and 3ab genes in vitro and in vivo.encephalomyocarditis virus (emcv) could infect many host species and cause acute myocarditis and sudden death in pre-weaned piglets. it was necessary to develop new antiviral strategies for the treatment of the virus infection. here, four plasmids expressing shrna (small hairpin rna) targeted to 1d or 3ab protein genes of emcv were constructed and their inhibition efficiency on the replication of emcv was evaluated in both bhk21 cells and mice. the results showed that three out of those four shr ...201122052534
A new splice variant of the human guanylate-binding protein 3 mediates anti-influenza activity through inhibition of viral transcription and replication.Guanylate-binding proteins (GBPs) belong to the family of large GTPases that are induced in response to interferons. GBPs contain an N-terminal globular GTPase domain and a C-terminal a-helical regulatory domain that are connected by a short middle domain. Antiviral activity against vesicular stomatitis virus and encephalomyocarditis virus has been shown for hGBP-1; however, no anti-influenza virus properties for GBPs have been described to date. Here we show that hGBP-1 and hGBP-3 possess anti- ...201122106366
SARS Coronavirus nsp1 Protein Induces Template-Dependent Endonucleolytic Cleavage of mRNAs: Viral mRNAs Are Resistant to nsp1-Induced RNA Cleavage.SARS coronavirus (SCoV) nonstructural protein (nsp) 1, a potent inhibitor of host gene expression, possesses a unique mode of action: it binds to 40S ribosomes to inactivate their translation functions and induces host mRNA degradation. Our previous study demonstrated that nsp1 induces RNA modification near the 5'-end of a reporter mRNA having a short 5' untranslated region and RNA cleavage in the encephalomyocarditis virus internal ribosome entry site (IRES) region of a dicistronic RNA template ...201122174690
latent hit series hidden in high-throughput screening data.recently a novel method termed compound set enrichment (cse) has been described that uses the activity distribution of a structural class of compounds to identify hit series from primary screening data. this report describes how this method can be used to identify such hit series, even when no hits according to conventional hit-calling methods for a given structural class are present in the data set. such series, which were called latent hit series, were identified prospectively in a cell-based ...201122185196
in vitro inhibition of influenza a virus infection by marine microalga-derived sulfated polysaccharide p-kg03.the sulfated polysaccharide, p-kg03, purified from the marine microalga, gyrodinium impudium, is a unique compound comprising homogenous galactose units conjugated to uronic acid and sulfated groups. although previous studies showed that p-kg03 suppresses tumor cell growth and infection by encephalomyocarditis virus, its effect against enveloped virus infection and the biological mechanism of action have not been elucidated. in this report, the inhibitory activity of p-kg03 against influenza vir ...201122197247
effects of magnesium ions on two emcv ires key activity fragments.using nmr magnetization transfer from water and ammonia-catalyzed exchange of the imino protons, changes have been monitored in base-pair kinetics induced by mg(2 + ) in two key activity fragments r(caccuggcgacaggug) and r(ggccaaaagcc) of the encephalomyocarditis virus internal ribosome entry site. for r(caccuggcgacaggug), the addition of mg(2 + ) reveals two types of base-pairs: r(u(545)·a) and r(g(546)·c), in the first category, have lifetimes only slightly higher in the presence of mg(2 + ), ...201122210961
a vaccine candidate for eastern equine encephalitis virus based on ires-mediated attenuation.to develop an effective vaccine against eastern equine encephalitis (eee), we engineered a recombinant eee virus (eeev) that was attenuated and capable of replicating only in vertebrate cells, an important safety feature for live vaccines against mosquito-borne viruses. the subgenomic promoter was inactivated with 13 synonymous mutations and expression of the eeev structural proteins was placed under the control of an internal ribosomal entry site (ires) derived from encephalomyocarditis virus ( ...201222222869
promotion of viral internal ribosomal entry site-mediated translation under amino acid starvation.cap-dependent and internal ribosomal entry site (ires)-mediated translation are regulated differently within cells. viral ires-mediated translation often remains active when cellular cap-dependent translation is severely impaired under cellular stresses induced by virus infection. to investigate how cellular stresses influence the efficiency of viral ires-mediated translation, we used a bicistronic luciferase reporter construct harbouring ires elements from the following viruses: encephalomyocar ...201222302880
inhibition of hepatitis b virus replication by phospholipid scramblase 1 in vitro and in vivo.human phospholipid scramblase 1 (plscr1) is an α/β interferon-inducible protein that mediates antiviral activity against rna viruses including vesicular stomatitis virus (vsv) and encephalomyocarditis virus (emcv). in the present study, we investigated the antiviral activity of plscr1 protein against hbv (hepatitis b virus). firstly, plscr1 mrna and protein expression was found to be downregulated in hepg2 cells after hbv infection. then by performing co-transient-transfection experiments in cel ...201222342889
l-tryptophan-kynurenine pathway metabolites regulate type i ifns of acute viral myocarditis in mice.the activity of ido that catalyzes the degradation of tryptophan (trp) into kynurenine (kyn) increases after diseases caused by different infectious agents. previously, we demonstrated that ido has an important immunomodulatory function in immune-related diseases. however, the pathophysiological role of ido following acute viral infection is not fully understood. to investigate the role of ido in the l-trp-kyn pathway during acute viral myocarditis, mice were infected with encephalomyocarditis v ...201222422885
target-size embracing dimension for sensitive detection of viruses with various sizes and influenza virus strains.the focused ion beam (fib) technique was employed to precisely fabricate hexagon-like au nano-rods (fibau_h) arrays as a surface enhanced raman scattering - active substrate. a "ring diameter" (d(r)) was created by the convergence of three fibau_h with respect to the dimension of the target viruses (d(t)), such as adenovirus (adeno), encephalomyocarditis virus (emcv), influenza virus (h1n1) with different sizes. three influenza a virus strains were also compared. the results indicate that as tha ...201222425238
[toll-like receptor dependent innate immune responses by primary mouse hepatocytes and its control of hbv replication].this report aims to investigate the toll-like receptor (tlr) signaling pathways and induced antiviral activity in hepatocytes.201122433306
mutagenesis-mediated virus extinction: virus-dependent effect of viral load on sensitivity to lethal defection.lethal mutagenesis is a transition towards virus extinction mediated by enhanced mutation rates during viral genome replication, and it is currently under investigation as a potential new antiviral strategy. viral load and virus fitness are known to influence virus extinction. here we examine the effect or the multiplicity of infection (moi) on progeny production of several rna viruses under enhanced mutagenesis.201222442668
a novel vaccine combined with an alum adjuvant for porcine encephalomyocarditis virus (emcv)-induced reproductive failure in pregnant sows.to evaluate a novel vaccine for porcine encephalomyocarditis virus (emcv), which causes reproductive failure in pregnant sows, virus like particles (vlps) were generated and immunized twice in 2 week intervals before sow mating. sows were divided into 4 groups (n=4, per group). group 1 was immunized with the alum adjuvant alone, group 2 with vlps alone, group 3 with vlps mixed alum adjuvant, and group 4 with a commercial killed vaccine. in group 2, seroconversion was observed at very low levels, ...201222445890
[construction and functional characterization of a monocistronic replicon based on the hcv genotype 2a promotor].to construct a hepatitis c virus (hcv) genotype 2a monocistronic replicon and investigate its replication capabilities in the human hepatocarcinoma cell lines, huh7.5 and huh7.1, in order to determine its potential as a molecular tool for future in vitro studies of hcv replication and selection studies for putative anti-hcv drugs. site-directed mutagenesis was used to delete the core-e1-e2-p7-ns2 fragment (about 3090 bp) from plasmid pj6jfh1blarl. the resultant trianglepj6jfh1blarl plasmid was d ...201222464780
preclinical studies of pf-04849285, an interferon-α8 fusion protein for the treatment of hcv.this study presents preclinical data of a novel interferon (ifn)-α8 fusion protein, pf-04849285, and compares it with ifn-α2 and pegylated ifn-α2; the latter being the current standard of care for hcv.201222505586
high susceptibility to lipopolysaccharide-induced lethal shock in encephalomyocarditis virus-infected mice.secondary bacterial infection in humans is one of the pathological conditions requiring clinical attention. in this study, we examined the effect of lipopolysaccharide (lps) on encephalomyocarditis virus (emcv) infected mice. all mice inoculated with emcv at 5 days before lps challenge died within 24 h. lps-induced tnf-α mrna expression was significantly increased in the brain and heart at 5 days after emcv infection. cd11b(+)/tlr4(+) cell population in the heart was remarkably elevated at 5 day ...201222509465
inhibition of type i interferon-mediated antiviral action in human glioma cells by the ikk inhibitors bms-345541 and tpca-1.the nuclear factor-kappa b (nfκb) signal transduction pathway plays an important role in immunity, inflammation, cell growth, and survival. since dysregulation of this pathway results in high, constitutive nfκb activation in various cancers and immune disorders, the development of specific drugs to target this pathway has become a focus for treating these diseases. nfκb regulates various aspects of the cellular response to interferon (ifn). however, the role of the upstream regulator of the nfκb ...201222509977
expression of biologically active human interferon alpha 2 in aloe vera.methods necessary for the successful transformation and regeneration of aloe vera were developed and used to express the human protein, interferon alpha 2 (ifnα2). ifnα2 is a secreted cytokine that plays a vital role in regulating the cellular response to viral infection. transgenic plants were regenerated from callus cultures initiated from zygotic embryos. expression of the ifna2 transgene in transformed plants was confirmed by rt-pcr and ifnα2 protein was detected by immunoblot analysis. huma ...201222528466
isolation, molecular characterization, and phylogenetic analysis of porcine encephalomyocarditis virus strain hb10 in china.encephalomyocarditis virus (emcv) can cause myocarditis, respiratory failure, reproductive failure, and mortality in pregnant sows, fetuses, and ablactating piglets. diseases caused by emcv currently affect the swine industry worldwide. a virus was isolated from organs of dead piglets that presented with acute myocarditis in northern china. the production of a specific cytopathic effect on susceptible cells and the results of hemagglutination inhibition (hi) assay, pcr, electron microscopy (em), ...201222538207
high throughput screening for small molecule enhancers of the interferon signaling pathway to drive next-generation antiviral drug discovery.most of current strategies for antiviral therapeutics target the virus specifically and directly, but an alternative approach to drug discovery might be to enhance the immune response to a broad range of viruses. based on clinical observation in humans and successful genetic strategies in experimental models, we reasoned that an improved interferon (ifn) signaling system might better protect against viral infection. here we aimed to identify small molecular weight compounds that might mimic this ...201222574190
interferon-induced ifit2/isg54 protects mice from lethal vsv neuropathogenesis.interferon protects mice from vesicular stomatitis virus (vsv) infection and pathogenesis; however, it is not known which of the numerous interferon-stimulated genes (isg) mediate the antiviral effect. a prominent family of isgs is the interferon-induced with tetratricopeptide repeats (ifit) genes comprising three members in mice, ifit1/isg56, ifit2/isg54 and ifit3/isg49. intranasal infection with a low dose of vsv is not lethal to wild-type mice and all three ifit genes are induced in the centr ...201222615570
trim22: a diverse and dynamic antiviral protein.the tripartite motif (trim) family of proteins is an evolutionarily ancient group of proteins with homologues identified in both invertebrate and vertebrate species. human trim22 is one such protein that has a dynamic evolutionary history that includes gene expansion, gene loss, and strong signatures of positive selection. to date, trim22 has been shown to restrict the replication of a number of viruses, including encephalomyocarditis virus (emcv), hepatitis b virus (hbv), and human immunodefici ...201222649727
the encephalomyocarditis virus.the encephalomyocarditis virus (emcv) is a small non-enveloped single-strand rna virus, the causative agent of not only myocarditis and encephalitis, but also neurological diseases, reproductive disorders and diabetes in many mammalian species. emcv pathogenesis appears to be viral strain- and host-specific, and a better understanding of emcv virulence factors is increasingly required. indeed, emcv is often used as a model for diabetes and viral myocarditis, and is also widely used in immunology ...201222722247
a real-time pcr to detect and analyze virulent emcv loads in sows and piglets.a real-time polymerase chain reaction with sybr green was developed for the detection and quantification of encephalomyocarditis virus (emcv) in porcine tissues; the method uses two primers specific for the 3d gene. the detection limit of this assay was 22 gene copies/reaction, equivalent to 0.001 tcid(50)/ml. the assay was linear over a 10(7) dilution range of template concentrations and was specific for emcv; it did not amplify other porcine pathogens (porcine circovirus 2, porcine reproductiv ...201222752806
protective efficacy of adenovirus-mediated small interfering rnas against encephalomyocarditis virus challenge in mice.encephalomyocarditis virus (emcv) infection can cause myocarditis and sudden death in pre-weaned piglets and severe reproductive failure in sows. there are no specific antiviral drugs for the treatment of the virus infection. in this study, four recombinant adenoviruses expressing small interfering rnas (sirnas) targeting to 1d or 3ab protein genes of emcv were constructed and their inhibition efficiency on the replication of emcv was evaluated in both marc-145 cells and mice. the results showed ...201222790206
antiviral activity of a small molecule deubiquitinase inhibitor occurs via induction of the unfolded protein response.ubiquitin (ub) is a vital regulatory component in various cellular processes, including cellular responses to viral infection. as obligate intracellular pathogens, viruses have the capacity to manipulate the ubiquitin (ub) cycle to their advantage by encoding ub-modifying proteins including deubiquitinases (dubs). however, how cellular dubs modulate specific viral infections, such as norovirus, is poorly understood. to examine the role of dubs during norovirus infection, we used wp1130, a small ...201222792064
isolation and molecular characterization of a second serotype of the encephalomyocarditis virus.encephalomyocarditis virus (emcv) and theilovirus are the two species of the cardiovirus genus. whereas theiloviruses comprise several sero-/genotypes, all known emcv isolates are serologically very similar and are thought to belong to one serotype, named emcv-1. here, a novel emcv type is described. strain rd 1338 (d28/05) was isolated from a wood mouse (apodemus sylvaticus) in germany and can be distinguished from emcv-1 by serological and molecular means. failure of emcv-1 specific hyperimmun ...201222824255
site-specific cleavage of the host poly(a) binding protein by the encephalomyocarditis virus 3c proteinase stimulates viral replication.although picornavirus rna genomes contain a 3'-terminal poly(a) tract that is critical for their replication, the impact of encephalomyocarditis virus (emcv) infection on the host poly(a)-binding protein (pabp) remains unknown. here, we establish that emcv infection stimulates site-specific pabp proteolysis, resulting in accumulation of a 45-kda n-terminal pabp fragment in virus-infected cells. expression of a functional emcv 3c proteinase was necessary and sufficient to stimulate pabp cleavage ...201222837200
role of microglia in oxidative toxicity associated with encephalomycarditis virus infection in the central nervous system.the single-stranded rna encephalomyocarditis virus (emcv) can replicate in the central nervous system (cns) and lead to prominent brain lesions in the stratum pyramidale hippocampus and the stratum granulosum cerebelli. activated microglia cells infected by emcv produce a massive burst of reactive oxygen species (ros) via nadph oxidase 2 (nox2) activation, leading to neuronal death. balancing this effect is mechanisms by which ros are eliminated from the cns. cellular prion protein (prp(c)) play ...201222837699
feeding of the probiotic bacterium enterococcus faecium ncimb 10415 differentially affects shedding of enteric viruses in pigs.effects of probiotic bacteria on viral infections have been described previously. here, two groups of sows and their piglets were fed with or without feed supplementation of the probiotic bacterium enterococcus faecium ncimb 10415. shedding of enteric viruses naturally occurring in these pigs was analyzed by quantitative real-time rt-pcr. no differences between the groups were recorded for hepatitis e virus, encephalomyocarditis virus and norovirus. in contrast, astrovirus was exclusively detect ...201222838386
identification of a germicidal compound against picornavirus in bamboo pyroligneous acid.the germicidal activity of pyroligneous acid (pa) against a picornavirus, encephalomyocarditis virus (emcv), was analyzed, and the component responsible for its disinfectant activity was identified. bamboo pa (bpa) inactivated emcv, but neutralization of bpa abolished this activity. using liquid-liquid phase extraction and silica gel column chromatography, the hydrophobic active fraction of bpa was separated and its 12 major components were identified. the active fraction was reconstructed by mi ...201222849724
comment on masek-hammerman et al, "epizootic myocarditis associated with encephalomyocarditis virus in a group of rhesus macaques (macaca mulatta)," vet pathol. 2012;49:386-392. 201322855414
rnase l triggers autophagy in response to viral infections.autophagy is a programmed homeostatic response to diverse types of cellular stress that disposes of long-lived proteins, organelles, and invading microbes within double-membraned structures called autophagosomes. the 2',5'-oligoadenylate/rnase l system is a virus-activated host rnase pathway that disposes of or processes viral and cellular single-stranded rnas. here we report that activation of rnase l during viral infections induces autophagy. accordingly, infections with encephalomyocarditis v ...201222875977
localized ires-dependent translation of er chaperone protein mrna in sensory axons.transport of neuronal mrnas into distal nerve terminals and growth cones allows axonal processes to generate proteins autonomous from the cell body. while the mechanisms for targeting mrnas for transport into axons has received much attention, how specificity is provided to the localized translational apparatus remains largely unknown. in other cellular systems, protein synthesis can be regulated by both cap-dependent and cap-independent mechanisms. the possibility that these mechanisms are used ...201222911708
encephalomyocarditis virus viroporin 2b activates nlrp3 inflammasome.nod-like receptors (nlrs) comprise a large family of intracellular pattern- recognition receptors. members of the nlr family assemble into large multiprotein complexes, termed the inflammasomes. the nlr family, pyrin domain-containing 3 (nlrp3) is triggered by a diverse set of molecules and signals, and forms the nlrp3 inflammasome. recent studies have indicated that both dna and rna viruses stimulate the nlrp3 inflammasome, leading to the secretion of interleukin 1 beta (il-1β) and il-18 follow ...201222916014
vaccination of llamas, llama glama, with an experimental killed encephalomyocarditis virus vaccine.encephalomyocarditis virus (emcv) is a pandemic virus that has caused mortality in numerous captive wildlife species worldwide. an experimental killed vaccine was created from two emcv isolates associated with zoo animal mortality in the southern united states. the vaccine was tested for safety and efficacy in eleven llamas (llama glama). all animals received an initial vaccination and a second booster vaccination 4 wk later. serum antibody responses were monitored at initial vaccination and at ...201122946372
retrospective genetic characterisation of encephalomyocarditis viruses from african elephant and swine recovers two distinct lineages in south africa.encephalomyocarditis virus (emcv) outbreaks are rare in southern africa. only two have been reported to date from south africa, both coinciding with rodent irruptions. the first outbreak manifested as acute myocarditis in pigs in 1979, whilst the second, occurring from 1993 to 1994, was linked to the deaths of 64 free-ranging adult african elephants (loxodonta africana). the p1 genome region, inclusive of the flanking leader (l) and 2a genes, of three south african isolates, one from swine and t ...201323021643
pathogenicity and molecular analysis of an encephalomyocarditis virus isolate from mideastern china.encephalomyocarditis virus (emcv) can infect many host species and cause acute myocarditis and sudden death in preweaned piglets. in this study, an emcv strain (nj08) was isolated from newborn pigs with clinical signs on a pig farm in mideastern china. it was identified by indirect immunofluorescence assay and reverse-transcription polymerase chain reaction. experiments showed that the isolate could cause severe clinical symptoms and pathological changes in mice but no obvious clinical and patho ...201223024459
pellino3 targets the irf7 pathway and facilitates autoregulation of tlr3- and viral-induced expression of type i interferons.toll-like receptors (tlrs) sense pathogen-associated molecules and respond by inducing cytokines and type i interferon. here we show that genetic ablation of the e3 ubiquitin ligase pellino3 augmented the expression of type i interferon but not of proinflammatory cytokines in response to tlr3 activation. pellino3-deficient mice had greater resistance against the pathogenic and lethal effects of encephalomyocarditis virus (emcv). tlr3 signaling induced pellino3, which in turn interacted with and ...201223042151
mrna decay during herpes simplex virus (hsv) infections: mutations that affect translation of an mrna influence the sites at which it is cleaved by the hsv virion host shutoff (vhs) protein.during lytic infections, the herpes simplex virus (hsv) virion host shutoff (vhs) endoribonuclease degrades many host and viral mrnas. within infected cells it cuts mrnas at preferred sites, including some in regions of translation initiation. vhs binds the translation initiation factors eif4h, eif4ai, and eif4aii, suggesting that its mrna degradative function is somehow linked to translation. to explore how vhs is targeted to preferred sites, we examined the in vitro degradation of a target mrn ...201323077305
genetic analysis of saffold virus-penang in relation to other newly discovered saffold viruses.viruses in the family picornaviridae are classified into nine genera. within the family picornaviridae, two species: encephalomyocarditis virus and theilovirus, are listed under the genus cardiovirus. a novel theilovirus, saffold virus (safv), was first reported in 2007. since then, numerous safv isolates have been detected around the world and genetic recombinations have been reported among them. in 2009, safv-penang was isolated from a febrile child with influenza-like illness in malaysia. saf ...201223077814
theiler's murine encephalomyelitis virus contrasts with encephalomyocarditis and foot-and-mouth disease viruses in its functional utilization of the stopgo non-standard translation mechanism.the picornaviruses' genome consists of a positive-sense ssrna. like many picornaviruses, cardioviruses synthesize two distinct polyprotein precursors from adjacent but non-overlapping genome segments. both the [l-1abcd-2a] and the [2bc-3abcd] polyproteins are proteolytically processed to yield mature capsid and non-structural proteins, respectively. an unusual translational event, known as 'stopgo' or 'stop-carry on', is responsible for the release of the [l-1abcd-2a] polyprotein from the riboso ...201323100365
rnase l induces autophagy via c-jun n-terminal kinase and double-stranded rna-dependent protein kinase signaling pathways.autophagy is a tightly regulated mechanism that mediates sequestration, degradation, and recycling of cellular proteins, organelles, and pathogens. several proteins associated with autophagy regulate host responses to viral infections. ribonuclease l (rnase l) is activated during viral infections and cleaves cellular and viral single-stranded rnas, including rrnas in ribosomes. here we demonstrate that direct activation of rnase l coordinates the activation of c-jun n-terminal kinase (jnk) and d ...201223109342
purification and visualization of encephalomyocarditisvirus synthesized by an in vitro protein expression system derived from mammalian cell extract.virus particles are promising vehicles and templates for vaccination, drug delivery and material sciences. although infectious picornaviruses can be synthesized from genomic or synthetic rna by cell-free protein expression systems derived from mammalian cell extract, there has been no direct evidence that authentic viral particles are indeed synthesized in the absence of living cells. we purified encephalomyocarditis virus (emcv) synthesized by a hela cell extract-derived, cell-free protein expr ...201323143176
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