Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
|---|
| microbiological survey of mice (mus musculus) purchased from commercial pet shops in kanagawa and tokyo, japan. | information regarding the prevalence of infectious agents in mice in pet shops in japan is scarce. this information is particularly useful for minimizing the risk of potential transmission of infections to laboratory mice. therefore, we surveyed infectious agents in mice from pet shops in kanagawa and tokyo, japan. the survey was conducted in 28 mice from 5 pet shops to screen for 47 items (17 viruses, 22 bacteria and fungi, 10 parasites) using culture tests, serology, pcr, and microscopy. the m ... | 2014 | 25502736 |
| t helper cell- and cd40-dependent germline igm prevents chronic virus-induced demyelinating disease. | generation of antiviral igm is usually considered as a marker of a short-lived initial antibody response that is replaced by hypermutated and more-efficient igg. however, once viruses have established a particular niche for their persistence (e.g., within the cns), the immune system has to specifically mobilize a broad range of antimicrobial effectors to contain the pathogen in the long term. infection of the cns with the mouse hepatitis virus (mhv) provides a unique model situation in which the ... | 2012 | 22232667 |
| receptor tyrosine kinase inhibitors that block replication of influenza a and other viruses. | we have previously reported that two receptor tyrosine kinase inhibitors (rtkis), called ag879 and tyrphostin a9 (a9), can each block the replication of influenza a virus in cultured cells. in this study, we further characterized the in vitro antiviral efficacies and specificities of these agents. the 50% effective concentration (ec(50)) of each against influenza a was found to be in the high nanomolar range, and the selectivity index (si = 50% cytotoxic concentration [cc(50)]/ec(50)) was determ ... | 2011 | 21930873 |
| severe gammaherpesvirus-induced pneumonitis and fibrosis in syngeneic bone marrow transplant mice is related to effects of transforming growth factor-β. | pulmonary infections and pneumonitis occur frequently after hematopoietic stem cell transplantation. using a syngeneic mouse model of bone marrow transplantation (bmt), we have previously demonstrated that bmt mice are more susceptible to acute gammaherpesvirus 68 (mhv-68) replication at day 7 after infection. by day 21, the virus is latent in lungs of bmt and control mice, and there is no difference in viral load. despite similar latent viral load, bmt mice develop severe pneumonitis associated ... | 2011 | 21924228 |
| surgical transplantation of mouse neural stem cells into the spinal cords of mice infected with neurotropic mouse hepatitis virus. | mice infected with the neurotropic jhm strain of mouse hepatitis virus (mhv) develop pathological and clinical outcomes similar to patients with the demyelinating disease multiple sclerosis (ms). we have shown that transplantation of nscs into the spinal cords of sick mice results in a significant improvement in both remyelination and in clinical outcome. cell replacement therapies for the treatment of chronic neurologic diseases are now a reality and in vivo models are vital in understanding th ... | 2011 | 21775959 |
| cyclosporin a inhibits the replication of diverse coronaviruses. | low micromolar, non-cytotoxic concentrations of cyclosporin a (csa) strongly affected the replication of sars-coronavirus (sars-cov), human coronavirus 229e, and mouse hepatitis virus in cell culture, as was evident from the strong inhibition of green fluorescent protein reporter gene expression and up to 4 log reduced progeny titres. upon high-multiplicity infection, csa treatment rendered sars-cov rna and protein synthesis almost undetectable, suggesting an early block in replication. sirna-me ... | 2011 | 21752960 |
| macrophage-mediated optic neuritis induced by retrograde axonal transport of spike gene recombinant mouse hepatitis virus. | after intracranial inoculation, neurovirulent mouse hepatitis virus (mhv) strains induce acute inflammation, demyelination, and axonal loss in the central nervous system. prior studies using recombinant mhv strains that differ only in the spike gene, which encodes a glycoprotein involved in virus-host cell attachment, demonstrated that spike mediates anterograde axonal transport of virus to the spinal cord. a demyelinating mhv strain induces optic neuritis, but whether this is due to the retrogr ... | 2011 | 21572336 |
| infectious microorganisms in mice (mus musculus) purchased from commercial pet shops in germany. | in this study, we investigated the prevalence of infectious microorganisms (viruses, bacteria, fungi and eukaryotic parasites) in mice from different pet shops in germany; such animals may compromise the hygienic integrity of laboratory animal vivaria if private pet holders act as unintended vectors of infections carried by them. house mice sold as pets or feed specimens were purchased from different pet shops and tested for a comprehensive panel of unwanted microorganisms. we found a number of ... | 2011 | 21508117 |
| use of lectins for characterization of o-linked glycans of herpes simplex virus glycoproteins. | the existence of o-linked glycans in viral glycoproteins was described in the early 1980s for enveloped viruses such as herpes simplex virus type 1 (hsv-1), vaccinia virus, and mouse hepatitis virus (1-4). glycoprotein c of hsv-1 (designated gc-1) was demonstrated to contain domains, in which numerous o-linked glycans were concentrated to pronase-resistant clusters (5-7), thereby resembling the organization of mucins (8). thus glycoprotein, containing nine sites for n-linked glycosylation in add ... | 1998 | 21374459 |
| autocrine interferon priming in macrophages but not dendritic cells results in enhanced cytokine and chemokine production after coronavirus infection. | coronaviruses efficiently inhibit interferon (ifn) induction in nonhematopoietic cells and conventional dendritic cells (cdc). however, ifn is produced in infected macrophages, microglia, and plasmacytoid dendritic cells (pdc). to begin to understand why ifn is produced in infected macrophages, we infected bone marrow-derived macrophages (bmm) and as a control, bone marrow-derived dc (bmdc) with the coronavirus mouse hepatitis virus (mhv). as expected, bmm but not bmdc expressed type i ifn. ifn ... | 2010 | 20978536 |
| murine coronavirus receptors are differentially expressed in the central nervous system and play virus strain-dependent roles in neuronal spread. | coronavirus infection of the murine central nervous system (cns) provides a model for studies of viral encephalitis and demyelinating disease. mouse hepatitis virus (mhv) neurotropism varies by strain: mhv-a59 causes mild encephalomyelitis and demyelination, while the highly neurovirulent strain jhm.sd (mhv-4) causes fatal encephalitis with extensive neuronal spread of virus. in addition, while neurons are the predominant cns cell type infected in vivo, the canonical receptor for mhv, the carcin ... | 2010 | 20739537 |
| migration of engrafted neural stem cells is mediated by cxcl12 signaling through cxcr4 in a viral model of multiple sclerosis. | multiple sclerosis (ms) is a human demyelinating disease characterized by multifocal regions of inflammation, progressive myelin loss within the central nervous system (cns), and eventual failure to remyelinate damaged axons. these problems suggest deficiencies in recruiting and/or maturation of oligodendrocyte progentior cells (opcs) and highlight cell replacement therapies to promote remyelination. we have used a model of viral-induced demyelination to characterize signaling cues associated wi ... | 2010 | 20534452 |
| the proteasome inhibitor velcade enhances rather than reduces disease in mouse hepatitis coronavirus-infected mice. | many viruses, including coronaviruses (covs), depend on a functional cellular proteasome for efficient infection in vitro. hence, the proteasome inhibitor velcade (bortezomib), a clinically approved anticancer drug, shown in an accompanying study (m. raaben et al., j. virol. 84:7869-7879, 2010) to strongly inhibit mouse hepatitis cov (mhv) infection in cultured cells, seemed an attractive candidate for testing its antiviral properties in vivo. surprisingly, however, the drug did not reduce repli ... | 2010 | 20484516 |
| the pathogenesis of murine coronavirus infection of the central nervous system. | mouse hepatitis virus (mhv) is a positive-strand rna virus that causes an acute encephalomyelitis that later resolves into a chronic fulminating demyelinating disease. cytokine production, chemokine secretion, and immune cell infiltration into the central nervous system are critical to control viral replication during acute infection. despite potent antiviral t-lymphocyte activity, sterile immunity is not achieved, and mhv chronically persists within oligodendrocytes. continued infiltration and ... | 2010 | 20370625 |
| pathogenesis of a model gammaherpesvirus in a natural host. | murine gammaherpesvirus 68 (mhv-68) infection of laboratory mice (mus musculus) is an established model of gammaherpesvirus pathogenesis. the fact that m. musculus is not a host in the wild prompted us to reassess mhv-68 infection in wood mice (apodemus sylvaticus), a natural host. here, we report significant differences in mhv-68 infection in the two species: (i) following intranasal inoculation, mhv-68 replicated in the lungs of wood mice to levels approximately 3 log units lower than in balb/ ... | 2010 | 20130062 |
| the murine coronavirus nucleocapsid gene is a determinant of virulence. | the murine coronavirus, mouse hepatitis virus (mhv) strain a59, causes acute encephalitis and chronic demyelinating disease as well as hepatitis in mice. the jhm strain (also called mhv-4 or jhm.sd) causes fatal encephalitis and only minimal hepatitis. previous analysis of chimeric recombinant mhvs in which the spike gene, encoding the protein that mediates viral entry and cell-to-cell fusion, was exchanged between jhm and a59 showed that the spike plays a major role in determining organ tropism ... | 2010 | 20007284 |
| increased killing of liver nk cells by fas/fas ligand and nkg2d/nkg2d ligand contributes to hepatocyte necrosis in virus-induced liver failure. | the role of liver nk cells in virus-induced severe viral hepatitis and, subsequently, hepatic failure is not well defined. in this study, we investigated the role of liver nk cells in the development of hepatocyte necrosis in fulminant hepatic failure (fhf) and acute-on-chronic liver failure (aclf) because of viral infection. a mouse model of fhf induced by murine hepatitis virus strain 3 (mhv-3) was used to study the role of liver nk cells. samples from patients with hepatitis b virus-related a ... | 2010 | 19949088 |
| infectious diseases in wild mice (mus musculus) collected on and around the university of pennsylvania (philadelphia) campus. | laboratory mice serve as important models in biomedical research. monitoring these animals for infections and infestations and excluding causative agents requires extensive resources. despite advancements in detection and exclusion over the last several years, these activities remain challenging for many institutions. the infections and infestations present in laboratory mouse colonies are well documented, but their mode of introduction is not always known. one possibility is that wild rodents l ... | 2009 | 19887025 |
| fine specificity of autoantibodies induced by mouse hepatitis virus a59. | we have shown that mice infected with mouse hepatitis virus a59 (mhv-a59) develop autoantibodies (autoab) to liver and kidney fumarylacetoacetate hydrolase (fah). the autoab recognized conformational as well as linear antigenic determinants in the enzyme, and the autoimmune response was not entirely restricted to molecular mimicry and/or epitope spreading. since the n- and c-terminal portions of the enzyme were the most reactive with autoab, the fine specificity of these ab was investigated. imm ... | 2009 | 19811085 |
| two-step conformational changes in a coronavirus envelope glycoprotein mediated by receptor binding and proteolysis. | the coronaviruses mouse hepatitis virus type 2 (mhv-2) and severe acute respiratory syndrome coronavirus (sars-cov) utilize proteases to enter host cells. upon receptor binding, the spike (s) proteins of both viruses are activated for membrane fusion by proteases, such as trypsin, present in the environment, facilitating virus entry from the cell surface. in contrast, in the absence of extracellular proteases, these viruses can enter cells via an endosomal pathway and utilize endosomal cathepsin ... | 2009 | 19706706 |
| microbial contaminations of laboratory mice and rats in taiwan from 2004 to 2007. | limited data are available on the pathogen status of contemporary rodent colonies in taiwan. here we summarized the rodent pathogen diagnostic records of the taiwan national laboratory animal center during a 4-y period that representing approximately 10% of the rodent colonies in taiwan. demand for pathogen diagnostic service increased continuously from 2004 to 2007, with a 20% increase each year. in 2007, more than 20% of the mouse colonies were positive for mouse parvovirus, mouse hepatitis vi ... | 2009 | 19653946 |
| a novel cre recombinase imaging system for tracking lymphotropic virus infection in vivo. | detection, isolation, and identification of individual virus infected cells during long term infection are critical to advance our understanding of mechanisms of pathogenesis for latent/persistent viruses. however, current approaches to study these viruses in vivo have been hampered by low sensitivity and effects of cell-type on expression of viral encoded reporter genes. we have designed a novel cre recombinase (cre)-based murine system to overcome these problems, and thereby enable tracking an ... | 2009 | 19652715 |
| cxcl10 and trafficking of virus-specific t cells during coronavirus-induced demyelination. | chronic expression of cxc chemokine ligand 10 (cxcl10) in the central nervous system (cns) following infection with the neurotropic jhm strain of mouse hepatitis virus (jhmv) is associated with an immune-mediated demyelinating disease. treatment of mice with anti-cxcl10 neutralizing antibody results in limited cd4+ t cell infiltration into the cns accompanied by a reduction in white matter damage. the current study determines the antigen-specificity of the t lymphocytes present during chronic di ... | 2009 | 19626487 |
| development of a chimeric dna-rna hammerhead ribozyme targeting sars virus. | severe acute respiratory syndrome (sars) is a severe pulmonary infectious disease caused by a novel coronavirus. to develop an effective and specific medicine targeting the sars-coronavirus (cov), a chimeric dna-rna hammerhead ribozyme was designed and synthesized using a sequence homologous with the mouse hepatitis virus (mhv). | 2009 | 19420961 |
| a serological survey to evaluate contemporary prevalence of viral agents and mycoplasma pulmonis in laboratory mice and rats in western europe. | to evaluate current prevalence rates of 24 viruses and of the bacterium mycoplasma pulmonis, the authors retrospectively surveyed serological data obtained from laboratory mice and rats housed in more than 100 western european institutions. serum samples were submitted to the authors' institution for testing between january 2007 and june 2008. the prevalence of an infection was defined as the percentage of tested samples that yielded positive results for a specific agent. in mice, the most commo ... | 2009 | 19384313 |
| target-dependent b7-h1 regulation contributes to clearance of central nervous system infection and dampens morbidity. | the neurotropic coronavirus jhm strain of mouse hepatitis virus persists in oligodendroglia despite the presence of virus-specific cd8 t cells. expression of programmed death 1 (pd-1) and b7-h1 were studied during acute and persistent infection to examine whether this negative regulatory mechanism contributes to cns viral persistence. the majority of cns-infiltrating cd8 t cells expressed pd-1, with the highest levels on virus-specific cd8 t cells. moreover, despite control of infectious virus, ... | 2009 | 19380790 |
| expression of newly identified secretory ceacam1(a) isoforms in the intestinal epithelium. | carcinoembryonic antigen-related cell adhesion molecule 1 (ceacam1) regulates intestinal immunological homeostasis. however, precise expression patterns of ceacam1 isoforms remain poorly understood in the intestinal epithelia. focusing on the small intestinal epithelium of balb/c mice, we identified three novel splice variants encoding ceacam1(a)-2, -2c1, and -4c1 by rt-pcr. ceacam1(a)-2, -2c1, and -4c1 demonstrated secretory properties by transfection experiments in vitro. among them, ceacam1(a ... | 2009 | 19358828 |
| age-dependent pathogenesis of murine gammaherpesvirus 68 infection of the central nervous system. | gammaherpesvirus infection of the central nervous system (cns) has been linked to various neurological diseases, including meningitis, encephalitis, and multiple sclerosis. however, little is known about the interactions between the virus and the cns in vitro or in vivo. murine gammaherpesvirus 68 (mhv-68 or (gamma)hv-68) is genetically related and biologically similar to human gammaherpesviruses, thereby providing a tractable animal model system in which to study both viral pathogenesis and rep ... | 2009 | 19214440 |
| type i ifn-mediated protection of macrophages and dendritic cells secures control of murine coronavirus infection. | the swift production of type i ifns is one of the fundamental aspects of innate immune responses against viruses. plasmacytoid dendritic cell-derived type i ifns are of prime importance for the initial control of highly cytopathic viruses such as the mouse hepatitis virus (mhv). the aim of this study was to determine the major target cell populations of this first wave of type i ifns. generation of bone marrow-chimeric mice expressing the type i ifn receptor (ifnar) on either hemopoietic or non- ... | 2009 | 19124753 |
| glycogen synthase kinase-3 regulates the phosphorylation of severe acute respiratory syndrome coronavirus nucleocapsid protein and viral replication. | coronavirus (cov) nucleocapsid (n) protein is a highly phosphorylated protein required for viral replication, but whether its phosphorylation and the related kinases are involved in the viral life cycle is unknown. we found the severe acute respiratory syndrome cov n protein to be an appropriate system to address this issue. using high resolution page analysis, this protein could be separated into phosphorylated and unphosphorylated isoforms. mass spectrometric analysis and deletion mapping show ... | 2009 | 19106108 |
| [construction of p-hfgl2shrna and its effect on hfgl2 expression in vitro]. | our previous studies have shown that an anti-sense plasmid to mouse fibrinogen like protein 2 (mfgl2) significantly reduced mfgl2 expression in vivo, markedly ameliorated inflammatory infiltration, fibrin deposition and hepatocyte necrosis, prolonged the survival time period and elevated the survival rate in balb/cj mice with murine hepatitis virus type 3 (mhv-3) induced fulminant hepatitis. this study was designed to explore the opportunity of rna interference technique in the inhibitory applic ... | 2008 | 19105940 |
| manipulation of the coronavirus genome using targeted rna recombination with interspecies chimeric coronaviruses. | targeted rna recombination has proven to be a powerful tool for the genetic engineering of the coronavirus genome, particularly in its 3' part. here we describe procedures for the generation of recombinant and mutant mouse hepatitis virus and feline infectious peritonitis virus. key to the two-step method is the efficient selection of recombinant viruses based on host cell switching. the first step consists of the preparation---using this selection principle--of an interspecies chimeric coronavi ... | 2008 | 19057874 |
| primary clearance of murine gammaherpesvirus 68 by pkctheta-/- cd8 t cells is compromised in the absence of help from cd4 t cells. | cd4 t cells are dispensable for acute control of murine gammaherpesvirus 68 (mhv-68) but are necessary for effective long-term control of the virus by cd8 t cells. in contrast, protein kinase c theta (pkctheta) is not essential for either acute or long-term viral control. however, we found that while either cd4 or cd8 t cells could mediate the clearance of mhv-68 from the lungs of pkctheta(+/+) mice, pkctheta(-/-) mice depleted of either subset failed to clear the virus. these data suggest that ... | 2008 | 18818318 |
| mouse hepatitis virus infection of the cns: a model for defense, disease, and repair. | viral infection of the central nervous system (cns) results in varied outcomes ranging from encephalitis, paralytic poliomyelitis or other serious consequences. one of the principal factors that directs the outcome of infection is the localized innate immune response, which is proceeded by the adaptive immune response against the invading viral pathogen. the role of the immune system is to contain and control the spread of virus within the cns, and paradoxically, this response may also be pathol ... | 2008 | 18508518 |
| cd4 t cells contribute to virus control and pathology following central nervous system infection with neurotropic mouse hepatitis virus. | replication of the neurotropic mouse hepatitis virus strain jhm (jhmv) is controlled primarily by cd8(+) t-cell effectors utilizing gamma interferon (ifn-gamma) and perforin-mediated cytotoxicity. cd4(+) t cells provide an auxiliary function(s) for cd8(+) t-cell survival; however, their direct contribution to control of virus replication and pathology is unclear. to examine a direct role of cd4(+) t cells in viral clearance and pathology, pathogenesis was compared in mice deficient in both perfo ... | 2008 | 18094171 |
| tumor necrosis factor alpha is not a pathogenic determinant in acute lethal encephalitis induced by a highly neurovirulent strain of mouse hepatitis virus. | to investigate the role of tumor necrosis factor alpha (tnfalpha) in the pathogenesis of acute viral encephalitis, tnfalpha-deficient mice were infected with a highly neurovirulent strain of mouse hepatitis virus, jhm, and compared with jhm-infected c57bl/6 mice as controls. all the jhm-infected mice had succumbed to infection by 6 days postinfection. the virus replication kinetics, histopathological changes and mrna expression levels of proinflammatory cytokines in the brain did not differ betw ... | 2008 | 18074094 |
| experimental coronavirus retinopathy (ecor): retinal degeneration susceptible mice have an augmented interferon and chemokine (cxcl9, cxcl10) response early after virus infection. | mouse hepatitis virus induces a biphasic disease in balb/c mice that consists of an acute retinitis followed by progression to a chronic retinal degeneration with autoimmune reactivity. retinal degeneration resistant cd-1 mice do not develop the late phase. what host factors contribute to the distinct responses to the virus are unknown. herein, we show that ifn-alpha, ifn-beta and ifn-gamma act in concert as part of the innate immune response to the retinal infection. at day 2, high serum levels ... | 2008 | 18037505 |
| myocarditis in newborn wild-type balb/c mice infected with the murine gamma herpesvirus mhv-68. | animal models of human epstein-barr virus (ebv) infection include ebv infection of primates and infection of mice with mhv-68, a further gamma herpesvirus (gamma-hv). we aimed at extending the mhv-68 model to study gamma-hv-related cardiac disease. | 2007 | 17658501 |
| cyclooxygenase activity is important for efficient replication of mouse hepatitis virus at an early stage of infection. | cyclooxygenases (coxs) play a significant role in many different viral infections with respect to replication and pathogenesis. here we investigated the role of coxs in the mouse hepatitis coronavirus (mhv) infection cycle. blocking cox activity by different inhibitors or by rna interference affected mhv infection in different cells. the cox inhibitors reduced mhv infection at a post-binding step, but early in the replication cycle. both viral rna and viral protein synthesis were affected with s ... | 2007 | 17555580 |
| new structure model for the packaging signal in the genome of group iia coronaviruses. | a 190-nucleotide (nt) packaging signal (ps) located in the 3' end of open reading frame 1b in the mouse hepatitis virus, a group iia coronavirus, was previously postulated to direct genome rna packaging. based on phylogenetic data and structure probing, we have identified a 95-nt hairpin within the 190-nt ps domain which is conserved in all group iia coronaviruses but not in the severe acute respiratory syndrome coronavirus (group iib), group i coronaviruses, or group iii coronaviruses. the hair ... | 2007 | 17428856 |
| a u-turn motif-containing stem-loop in the coronavirus 5' untranslated region plays a functional role in replication. | the 5' untranslated region (utr) of the mouse hepatitis virus (mhv) genome contains cis-acting sequences necessary for transcription and replication. a consensus secondary structural model of the 5' 140 nucleotides of the 5' utrs of nine coronaviruses (covs) derived from all three major cov groups is presented and characterized by three major stem-loops, sl1, sl2, and sl4. nmr spectroscopy provides structural support for sl1 and sl2 in three group 2 covs, including mhv, bcov, and hcov-oc43. sl2 ... | 2007 | 17353353 |
| membrane topology of murine coronavirus replicase nonstructural protein 3. | mouse hepatitis virus (mhv) is a member of the family coronaviridae. these positive strand rna viruses encode a replicase polyprotein that is processed into 16 nonstructural proteins (nsps). the nsps assemble with membranes to generate double membrane vesicles, which are the sites of viral rna synthesis. mhv nsp3 contains multiple domains including two papain-like protease domains, plp1 and plp2, and a predicted transmembrane (tm) domain. in this study, we determined the membrane topology of nsp ... | 2007 | 17222884 |
| murine coronavirus-induced oligodendrocyte apoptosis is mediated through the activation of the fas signaling pathway. | we previously showed that infection of rat oligodendrocytes by ultraviolet light-inactivated mouse hepatitis virus (mhv) resulted in apoptosis, suggesting that the apoptosis is triggered during cell entry. to further characterize the earliest apoptotic signaling events, here we treated cells with an antibody specific to the mhv receptor prior to and during virus infection or with an antibody specific to mhv spike protein following virus binding. both treatments blocked virus infection and apopto ... | 2007 | 17156812 |
| a hypervariable region within the 3' cis-acting element of the murine coronavirus genome is nonessential for rna synthesis but affects pathogenesis. | the 3' cis-acting element for mouse hepatitis virus (mhv) rna synthesis resides entirely within the 301-nucleotide 3' untranslated region (3' utr) of the viral genome and consists of three regions. encompassing the upstream end of the 3' utr are a bulged stem-loop and an overlapping rna pseudoknot, both of which are essential to mhv and common to all group 2 coronaviruses. at the downstream end of the genome is the minimal signal for initiation of negative-strand rna synthesis. between these two ... | 2007 | 17093194 |
| identification of the receptor for fgl2 and implications for susceptibility to mouse hepatitis virus (mhv-3)-induced fulminant hepatitis. | 2006 | 17037572 | |
| cd8+ t-cell priming during a central nervous system infection with mouse hepatitis virus. | 2006 | 17037564 | |
| receptor-independent infection of mouse hepatitis virus: analysis by spinoculation. | 2006 | 17037555 | |
| the nsp2 proteins of mouse hepatitis virus and sars coronavirus are dispensable for viral replication. | the results presented here demonstrate that the mhv and sars-cov nsp2 proteins are not required for the production of infectious virus, for polyprotein expression or processing, or for viral replication complex formation in cell culture. the nsp2 protein domain resides in a region of the coronavirus replicase that is relatively nonconserved across coronaviruses. in fact, the size and amino acid sequence variability of nsp2 across the different coronaviruses has led some investigators to speculat ... | 2006 | 17037506 |
| control of coronavirus infection through plasmacytoid dendritic-cell-derived type i interferon. | this study demonstrates a unique and crucial role of plasmacytoid dendritic cells (pdcs) and pdc-derived type i interferons (ifns) in the pathogenesis of mouse coronavirus infection. pdcs controlled the fast replicating mouse hepatitis virus (mhv) through the immediate production of type i ifns. recognition of mhv by pdcs was mediated via tlr7 ensuring a swift ifn-alpha production following encounter with this cytopathic rna virus. furthermore, the particular type i ifn response pattern was not ... | 2007 | 16985170 |
| cooperative involvement of the s1 and s2 subunits of the murine coronavirus spike protein in receptor binding and extended host range. | to study the process of spike (s)-receptor interaction during coronavirus entry, we evaluated the contributions of mutations in different regions of the murine hepatitis virus (mhv) s protein to natural receptor murine carcinoembryonic antigen-related cell adhesion molecule 1a (ceacam1a) dependence and to the acquisition of extended host range. extended-host-range variants of mhv strain a59 were previously obtained from persistently infected cells (j. h. schickli, b. d. zelus, d. e. wentworth, s ... | 2006 | 16956938 |
| both spike and background genes contribute to murine coronavirus neurovirulence. | various strains of mouse hepatitis virus (mhv) exhibit different pathogenic phenotypes. infection with the a59 strain of mhv induces both encephalitis and hepatitis, while the highly neurovirulent jhm strain induces a fatal encephalitis with little, if any, hepatitis. the pathogenic phenotype for each strain is determined by the genetic composition of the viral genome, as well as the host immune response. using isogenic recombinant viruses with a59 background genes differing only in the spike ge ... | 2006 | 16809289 |
| ultrastructure and origin of membrane vesicles associated with the severe acute respiratory syndrome coronavirus replication complex. | the rna replication complexes of mammalian positive-stranded rna viruses are generally associated with (modified) intracellular membranes, a feature thought to be important for creating an environment suitable for viral rna synthesis, recruitment of host components, and possibly evasion of host defense mechanisms. here, using a panel of replicase-specific antisera, we have analyzed the earlier stages of severe acute respiratory syndrome coronavirus (sars-cov) infection in vero e6 cells, in parti ... | 2006 | 16731931 |
| functional diversity of chemokines and chemokine receptors in response to viral infection of the central nervous system. | encounters with neurotropic viruses result in varied outcomes ranging from encephalitis, paralytic poliomyelitis or other serious consequences to relatively benign infection. one of the principal factors that control the outcome of infection is the localized tissue response and subsequent immune response directed against the invading toxic agent. it is the role of the immune system to contain and control the spread of virus infection in the central nervous system (cns), and paradoxically, this r ... | 2006 | 16570854 |
| differential roles for cxcr3 in cd4+ and cd8+ t cell trafficking following viral infection of the cns. | lymphocyte infiltration into the central nervous system (cns) following viral infection represents an important component of host defense and is required for control of viral replication. however, the mechanisms governing inflammation in response to viral infection of the cns are not well understood. following intracranial (i.c.) infection of susceptible mice with mouse hepatitis virus (mhv), mice develop an acute encephalomyelitis followed by a chronic demyelinating disease. the cxc chemokine l ... | 2006 | 16479546 |
| fibrinogen-like protein 2 fibroleukin expression and its correlation with disease progression in murine hepatitis virus type 3-induced fulminant hepatitis and in patients with severe viral hepatitis b. | to evaluate the expression of fibrinogen-like protein 2 (fgl2) and its correlation with disease progression in both mice and patients with severe viral hepatitis. | 2005 | 16437596 |
| redirecting coronavirus to a nonnative receptor through a virus-encoded targeting adapter. | murine hepatitis coronavirus (mhv)-a59 infection depends on the interaction of its spike (s) protein with the cellular receptor mceacam1a present on murine cells. human cells lack this receptor and are therefore not susceptible to mhv. specific alleviation of the tropism barrier by redirecting mhv to a tumor-specific receptor could lead to a virus with appealing properties for tumor therapy. to demonstrate that mhv can be retargeted to a nonnative receptor on human cells, we produced bispecific ... | 2006 | 16415002 |
| effect of mutations in the mouse hepatitis virus 3'(+)42 protein binding element on rna replication. | the mouse hepatitis virus (mhv) genome's 3' untranslated region contains cis-acting sequences necessary for replication. studies of mhv and other coronaviruses have indicated a role for rna secondary and tertiary elements in replication. previous work in our laboratory has identified four proteins which form ribonucleoprotein complexes with the 3'-terminal 42 nucleotides [3'(+)42] of the mhv genome. defective interfering (di) rna replication assays have demonstrated a role for the 3'(+)42 host p ... | 2005 | 16282457 |
| astrocyte expression of a dominant-negative interferon-gamma receptor. | interferon-gamma (ifn-gamma) is a major proinflammatory cytokine, and binding to its nearly ubiquitous receptor induces a wide variety of biological functions. to explore the role(s) of ifn-gamma signaling in astrocytes, transgenic mice (gfap/ifn-gammar1deltaic) expressing a dominant-negative ifn-gamma receptor alpha chain under control of the astrocyte-specific glial fibrillary acid protein (gfap) promoter were generated. transgenic mice developed normally, had normal astrocyte numbers and dist ... | 2005 | 16118798 |
| viral expression of ccl2 is sufficient to induce demyelination in rag1-/- mice infected with a neurotropic coronavirus. | mouse hepatitis virus strain jhm causes a chronic demyelinating disease in susceptible strains of rodents. demyelination does not develop in infected rag1-/- (recombination activation gene-deficient) mice but can be induced by several experimental interventions, including adoptive transfer of virus-specific t cells or antibodies. a common feature of demyelination in these models is extensive infiltration of macrophages/microglia into the white matter. the data obtained thus far do not indicate w ... | 2005 | 15890951 |
| the nucleocapsid protein of sars coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleoprotein a1. | the nucleocapsid (n) protein of sars coronavirus (sars_cov) is a major structural component of virions, which appears to be a multifunctional protein involved in viral rna replication and translation. heterogeneous nuclear ribonucleoprotein a1 (hnrnp a1) is related to the pre-mrna splicing in the nucleus and translation regulation in the cytoplasm. in this report, based on the relevant biophysical and biochemical assays, the nucleocapsid protein of sars_cov (sars_n) was discovered to exhibit hig ... | 2005 | 15862300 |
| transcriptome profile within the mouse central nervous system and activation of myelin-reactive t cells following murine coronavirus infection. | multiple sclerosis (ms) is an autoimmune disease associated with environmental factors, possibly including several viruses such as the coronaviruses. indeed, murine coronavirus (mhv) infection provides a well-known experimental model for ms studies. intracerebral infection of c57bl/6 mice with mhv-a59 revealed that viral replication was efficient and that clearance of infectious virus occurred as soon as 7 days post-infection. using cdna arrays, analysis of gene expression profile in the brain r ... | 2005 | 15833360 |
| maintenance of cd8+ t cells during acute viral infection of the central nervous system requires cd4+ t cells but not interleukin-2. | the jhm strain of mouse hepatitis virus (jhmv) is rapidly cleared from the central nervous system (cns) by cd8(+) t cells. in the absence of cd4(+) t cells, fewer cd8(+) t cells are found within the cns in association with a coordinate increase in apoptotic lymphocytes. previous data suggested that cd4(+) t cells may support cd8(+) t cells through secretion of interleukin-2 (il-2). to determine the in vivo role of il-2 during cns infection, il-2 signaling was inhibited via administration of a ne ... | 2005 | 15802960 |
| increased epitope-specific cd8+ t cells prevent murine coronavirus spread to the spinal cord and subsequent demyelination. | cd8+ t cells are important for clearance of neurotropic mouse hepatitis virus (mhv) strain a59, although their possible role in a59-induced demyelination is not well understood. we developed an adoptive-transfer model to more clearly elucidate the role of virus-specific cd8+ t cells during the acute and chronic phases of infection with a59 that is described as follows. c57bl/6 mice were infected with a recombinant a59 virus expressing the gp33 epitope, an h-2db-restricted cd8+ t-cell epitope enc ... | 2005 | 15731231 |
| pathogenesis of enterotropic mouse hepatitis virus in immunocompetent and immunodeficient mice. | mouse hepatitis virus (mhv) is one of the most prevalent viruses infecting laboratory mice. most natural infections are caused by enterotropic strains. experiments were done to compare the pathogenesis of enterotropic strain mhv-y in immunocompetent balb/c and c57bl/6 mice with that in b and t cell-deficient mice. in situ hybridization was used to identify sites of virus replication, and reverse transcriptase-polymerase chain reaction analysis was used to detect viral rna in feces and blood. mhv ... | 2004 | 15679267 |
| expression of cellular oncogene bcl-xl prevents coronavirus-induced cell death and converts acute infection to persistent infection in progenitor rat oligodendrocytes. | murine coronavirus mouse hepatitis virus (mhv) causes persistent infection of the central nervous system (cns) in rodents, which has been associated with demyelination. however, the precise mechanism of mhv persistence in the cns remains elusive. here we show that the progenitor oligodendrocytes (central glial 4 [cg-4] cells) derived from newborn rat brain were permissive to mhv infection, which resulted in cell death, although viral replication was restricted. interestingly, treatment with feta ... | 2005 | 15596800 |
| multi-phase approach to eradicate enzootic mouse coronavirus infection. | infections with mouse coronavirus (also known as mouse hepatitis virus, mhv) are common and prompt concern because the adverse research effects of infection have been well documented. the animal facility we describe had contained an enzootic infection of mouse coronavirus for more than a decade. eradication of the virus had been tried with limited success in the past. with an increase in the populations of immune-compromised and transgenic animals elsewhere in the facility, eradication of the vi ... | 2004 | 15461436 |
| recombinant viral sialate-o-acetylesterases. | viral o-acetylesterases were first identified in several viruses, including influenza c viruses and coronaviruses. these enzymes are capable of removing cellular receptors from the surface of target cells. hence they are also known as "receptor destroying" enzymes. we have cloned and expressed several recombinant viral o-acetylesterases. these enzymes were secreted from sf9 insect cells as chimeric proteins fused to egfp. a purification scheme to isolate the recombinant o-acetylesterase of influ ... | 2004 | 15454694 |
| mouse antibody production test: can we do without it? | introduction of microbiologically contaminated materials into mice can cause infections of the recipients and jeopardize experimental protocols. as such, the methods used to screen biological materials should be sensitive, reliable and suitable for routine diagnostic work. in this report, the sensitivity of the viral plaque assay, mouse antibody production test and polymerase chain reaction (pcr) for detection of mhv-a59 and mmvp, two of the most prevalent pathogenic viruses in experimental mous ... | 2004 | 15288967 |
| coronavirus spike glycoprotein, extended at the carboxy terminus with green fluorescent protein, is assembly competent. | due to the limited ultrastructural information about the coronavirion, little is known about the interactions acting at the interface between nucleocapsid and viral envelope. knowing that subtle mutations in the carboxy-terminal endodomain of the m protein are already lethal, we have now probed the equivalent domain of the spike (s) protein by extending it terminally with a foreign sequence of 27 kda: the green fluorescent protein (gfp). when expressed individually in murine cells, the s-gfp chi ... | 2004 | 15220410 |
| murine viral hepatitis involves nk cell depletion associated with virus-induced apoptosis. | mouse hepatitis virus type 3 (mhv3), a coronavirus, is an excellent animal model for the study of immunological disorders related to acute and chronic hepatitis. in this study, we have verified if the fulminant hepatitis induced by mhv3 could be related to an impairment of innate immunity. groups of three c57bl/6 mice were infected with the pathogenic l2-mhv3 or attenuated yac-mhv3 viruses, and the natural killer (nk) cell populations from liver, spleen and bone marrow were analysed. the percent ... | 2004 | 15196242 |
| cleavage between replicase proteins p28 and p65 of mouse hepatitis virus is not required for virus replication. | the p28 and p65 proteins of mouse hepatitis virus (mhv) are the most amino-terminal protein domains of the replicase polyprotein. cleavage between p28 and p65 has been shown to occur in vitro at cleavage site 1 (cs1), (247)gly downward arrow val(248), in the polyprotein. although critical residues for cs1 cleavage have been mapped in vitro, the requirements for cleavage have not been studied in infected cells. to define the determinants of cs1 cleavage and the role of processing at this site dur ... | 2004 | 15140993 |
| requirements for ceacams and cholesterol during murine coronavirus cell entry. | previous reports have documented that cholesterol supplementations increase cytopathic effects in tissue culture and also intensify in vivo pathogenicities during infection by the enveloped coronavirus murine hepatitis virus (mhv). to move toward a mechanistic understanding of these phenomena, we used growth media enriched with methyl-beta-cyclodextrin or cholesterol to reduce or elevate cellular membrane sterols, respectively. cholesterol depletions reduced plaque development 2- to 20-fold, dep ... | 2004 | 14990688 |
| very diverse cd8 t cell clonotypic responses after virus infections. | we measured cd8 t cell clonotypic diversity to three epitopes recognized in c57bl/6 mice infected with mouse hepatitis virus, strain jhm, or lymphocytic choriomeningitis virus. we isolated epitope-specific t cells with an ifn-gamma capture assay or mhc class i/peptide tetramers and identified different clonotypes by vbeta chain sequence analysis. in agreement with our previous results, the number of different clonotypes responding to all three epitopes fit a log-series distribution. from these d ... | 2004 | 14978121 |
| distinct roles for ip-10/cxcl10 in three animal models, theiler's virus infection, eae, and mhv infection, for multiple sclerosis: implication of differing roles for ip-10. | theiler's murine encephalomyelitis virus (tmev) causes demyelination with inflammation of the central nervous system (cns) in mice and is used as an animal model for multiple sclerosis (ms). interferon-gamma inducible protein-10 kda (ip-10) is a cxc chemokine and a chemoattractant for cxcr3+ t cells. ip-10 mrna is expressed in the cns during tmev infection. however, administration of anti-ip-10 serum caused no difference in clinical signs, inflammation, demyelination, virus persistence or anti-v ... | 2004 | 14760949 |
| coronavirus replication complex formation utilizes components of cellular autophagy. | the coronavirus mouse hepatitis virus (mhv) performs rna replication on double membrane vesicles (dmvs) in the cytoplasm of the host cell. however, the mechanism by which these dmvs form has not been determined. using genetic, biochemical, and cell imaging approaches, the role of autophagy in dmv formation and mhv replication was investigated. the results demonstrated that replication complexes co-localize with the autophagy proteins, microtubule-associated protein light-chain 3 and apg12. mhv i ... | 2004 | 14699140 |
| coronavirus-induced demyelination occurs in the absence of cd28 costimulatory signals. | infection of mice with mouse hepatitis virus (mhv) strain a59 results in acute encephalitis, hepatitis, and chronic demyelinating disease. t lymphocytes play an important role in mhv infection, and costimulatory signals are an important component of t cell function. to elucidate the role of the main costimulatory molecule, cd28, in mhv pathogenesis and demyelination, we examined the kinetics of mhv-a59 infection in cd28 knockout mice. mhv-a59-infected cd28 knockout mice developed acute encephali ... | 2004 | 14698856 |
| n-terminal domain of the murine coronavirus receptor ceacam1 is responsible for fusogenic activation and conformational changes of the spike protein. | the mouse hepatitis virus (mhv) receptor (mhvr), ceacam1, has two different functions for mhv entry into cells: binding to mhv spike protein (s protein) and activation of the s protein to execute virus-cell membrane fusion, the latter of which is accompanied by conformational changes of the s protein. the mhvr comprising the n-terminal and fourth domains [r1(1,4)] displays these two activities, and the n domain is thought to be critical for binding to mhv. in this study, we have addressed whethe ... | 2004 | 14671103 |
| induction of caspase-dependent apoptosis in cultured rat oligodendrocytes by murine coronavirus is mediated during cell entry and does not require virus replication. | murine coronavirus mouse hepatitis virus (mhv) causes demyelination of the central nervous system (cns) in rats and mice. apoptotic oligodendrocytes have been detected in the vicinity of the cns demyelinating lesions in these animals. however, whether mhv can directly induce oligodendrocyte apoptosis has not been documented. here, we established a rat oligodendrocyte culture that is morphologically and phenotypically indistinguishable from the primary rat oligodendrocytes. using this culture, we ... | 2003 | 14581532 |
| [metabolic aspects of hepatic tissue in mhv-3 virus infections of the mouse. ii. gama-glycerophosphate-dehydrogenase and pyruvic-kinase]. | 1961 | 14494229 | |
| multiplication and cytopathogenicity of mouse hepatitis virus in mouse cell cultures. | 1961 | 14476514 | |
| [new isolation of a mouse hepatitis virus (edp virus). its virological biological characteristics]. | 1960 | 14419911 | |
| an electron microscope study of the development of a mouse hepatitis virus in tissue culture cells. | samples taken at different intervals of time from suspension cultures of the nctc 1469 line of mouse liver-derived (ml) cells infected with a mouse hepatitis virus have been studied with the electron microscope. the experiments revealed that the viruses are incorporated into the cells by viropexis within 1 hour after being added to the culture. an increasing number of particles are found later inside dense cytoplasmic corpuscles similar to lysosomes. in the cytoplasm of the cells from the sample ... | 1965 | 14286297 |
| potentiating effect of k-virus on mouse hepatitis virus (mhv-s) in weanling mice. | 1963 | 14120346 | |
| mouse hepatitis virus infection as a highly contagious, prevalent, enteric infection of mice. | 1963 | 13982823 | |
| plaque formation by a mouse hepatitis virus. | 1962 | 13951824 | |
| [influence of diisopropylammonium dichloroacetate (diedi) in experimental mhv-3 virus (mouse hepatitis virus) infection]. | 1960 | 13781410 | |
| the enhancing effect of murine hepatitis virus on the cerebral activity of pleuropneumonia-like organisms in mice. | pleuropneumoma-like organisms (pplo) of the catarrhal type were isolated from the brain of a swiss mouse during the cranial passage of mouse hepatitis virus-mhv(c). cranial injection of the pplo alone in swiss and princeton weanlings was attended by a meagre growth of the organisms in the brain, with no pathologic change. the growth of both catarrhal and conjunctival strains of pplo in the brains of swiss mice was greatly enhanced by the simultaneous injection of mhv(c). rolling was not a charac ... | 1957 | 13449230 |
| the nature of mouse hepatitis virus infection in weanling mice. | 1955 | 13243199 | |
| multiple type a/b heterogeneous nuclear ribonucleoproteins (hnrnps) can replace hnrnp a1 in mouse hepatitis virus rna synthesis. | heterogeneous nuclear ribonucleoprotein (hnrnp) a1 has previously been shown to bind mouse hepatitis virus (mhv) rna at the 3' end of both plus and minus strands and modulate mhv rna synthesis. however, a mouse erythroleukemia cell line, cb3, does not express hnrnp a1 but still supports mhv replication, suggesting that alternative proteins can replace hnrnp a1 in cellular functions and viral infection. in this study, we set out to identify these proteins. uv cross-linking experiments revealed th ... | 2003 | 12970443 |
| stat1 and stat3 alpha/beta splice form activation predicts host responses in mouse hepatitis virus type 2 infection. j med virol 2003;69:306-312. | 2003 | 12966557 | |
| the glycosylation status of the murine hepatitis coronavirus m protein affects the interferogenic capacity of the virus in vitro and its ability to replicate in the liver but not the brain. | the coronavirus m protein, the most abundant coronaviral envelope component, is invariably glycosylated, which provides the virion with a diffuse, hydrophilic cover on its outer surface. remarkably, while the group 1 and group 3 coronaviruses all have m proteins with n-linked sugars, the m proteins of the group 2 coronaviruses [e.g., mouse hepatitis virus (mhv)] are o-glycosylated. the conservation of the n- and o-glycosylation motifs suggests that each of these types of carbohydrate modificatio ... | 2003 | 12919744 |
| [the study of cis-element hnf4 in the regulation of mfg12 prothrombinase/fibroleukin gene expression in response to nucleocapsid protein of mhv-3]. | to identify the transcription factor(s) that is essential for activation of mfgl2 prothrombinase/fibroleukin gene in response to nucleocapsid protein of murine hepatitis virus type 3 (mhv-3). | 2003 | 12887828 |
| control of central nervous system viral persistence by neutralizing antibody. | replication of the neurotropic jhm strain of mouse hepatitis virus within the central nervous system is controlled by cellular immunity. however, following initial clearance, virus reactivates in the absence of humoral immunity. viral recrudescence is prevented by the transfer of antiviral antibody (ab). to characterize the specificity and biological functions of ab critical for maintaining viral persistence, monoclonal abs specific for the viral spike, matrix, and nucleocapsid proteins were tra ... | 2003 | 12663773 |
| cloning of herpesviral genomes as bacterial artificial chromosomes. | herpesviruses, which are important pathogens for both animals and humans, have large and complex genomes with a coding capacity for up to 225 open reading frames (orfs). due to the large genome size and the slow replication kinetics in vitro of some herpesviruses, mutagenesis of viral genes in the context of the viral genome by conventional recombination methods in cell culture has been difficult. given that mutagenesis of viral genes is the basic strategy to investigate function, many of the he ... | 2003 | 12627394 |
| neutrophils promote mononuclear cell infiltration during viral-induced encephalitis. | neutrophils are the first infiltrating cell population to appear within the cns during infection with the neurotropic jhm strain of mouse hepatitis virus (jhmv). to determine whether neutrophils play a role in limiting acute jhmv infection, mice were depleted of neutrophils. infection of neutropenic animals resulted in increased levels of virus replication and mortality compared with control mice. furthermore, neutropenia resulted in significantly reduced mononuclear leukocyte infiltration possi ... | 2003 | 12626593 |
| the n-terminal domain of the murine coronavirus spike glycoprotein determines the ceacam1 receptor specificity of the virus strain. | using isogenic recombinant murine coronaviruses expressing wild-type murine hepatitis virus strain 4 (mhv-4) or mhv-a59 spike glycoproteins or chimeric mhv-4/mhv-a59 spike glycoproteins, we have demonstrated the biological functionality of the n-terminus of the spike, encompassing the receptor binding domain (rbd). we have used two assays, one an in vitro liposome binding assay and the other a tissue culture replication assay. the liposome binding assay shows that interaction of the receptor wit ... | 2003 | 12502800 |
| the art of survival during viral persistence. | central nervous system infection by the neurotropic jhm strain of mouse hepatitis virus (jhmv) results in chronic demyelination characterized by viral persistence in the absence of infectious virus. cd8(+) t cells inhibit acute viral replication via cell type-specific effector mechanisms. perforin-mediated cytolysis controls virus in microglia/macrophages and astrocytes, whereas interferon (ifn)-gamma regulates viral replication in oligodendroglia. jhmv infection of antibody-deficient mice confi ... | 2002 | 12491152 |
| receptor-induced conformational changes of murine coronavirus spike protein. | although murine coronavirus mouse hepatitis virus (mhv) enters cells by virus-cell membrane fusion triggered by its spike (s) protein, it is not well known how the s protein participates in fusion events. we reported that the soluble form of mhv receptor (somhvr) transformed a nonfusogenic s protein into a fusogenic one (f. taguchi and s. matsuyama, j. virol. 76:950-958, 2002). in the present study, we demonstrate that somhvr induces the conformational changes of the s protein, as shown by the p ... | 2002 | 12414924 |
| detection of mouse hepatitis virus infection by assay of anti-liver autoantibodies. | the observation that mice infected with mouse hepatitis virus (mhv) develop autoantibodies directed mainly to liver fumarylacetoacetate hydrolase (fah) enabled the development of an elisa applicable to the detection of mhv-infection. the method, based on the titration of antibodies to semipurified fah from rat liver, is easy, economical, and does not require the isolation of viral proteins from large mhv stocks. furthermore, since sera from mice immunized with a purified fraction of the rat live ... | 2002 | 12393146 |
| virus-induced demyelination in nude mice is mediated by gamma delta t cells. | infection of mice with mouse hepatitis virus (mhv), strain jhm, results in acute and chronic demyelination with many similarities to the human disease multiple sclerosis. this pathological process is primarily t cell-mediated and mhv infection of mice lacking b and t cells does not result in demyelination. in apparent contradiction to these results, robust demyelination is detected in mhv-infected young nude (athymic) mice. herein, we show that demyelination in nude mice was mediated by gamma de ... | 2002 | 12368199 |