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establishment of cell lines persistently infected with foot-and-mouth disease virus.cell lines persistently infected with foot-and-mouth disease virus (fmdv) have been established by growth of bhk-21 (c-13) or ibrs-2 (c-26) that survived standard cytolytic infections with fmdv. they maintain cytoplasmic fmdv rna sequences, as shown by dot blot hybridization tests, using cloned fmdv cdna as probes. cell line c1-bhk-rc1 was derived by infection of cloned bhk-21 c1 cells and plaque-purified fmdv c-s8 c1. indirect immunofluorescence assays indicated the presence of fmdv antigens. i ...19852990100
in vivo antiviral activity of recombinant murine gamma interferon.recombinant murine gamma interferon (gamma-ifn) was tested for its antiviral activity in vivo. ifn preparations purified to greater than 95% purity were administered to cd-1 mice infected with lethal doses of encephalomyocarditis (emc) virus. an initial treatment with rmuifn-gamma administered 4 h prior to infection with virus, followed by daily treatment for 3 consecutive days significantly protected mice against emc virus as evidenced by animal survival after 3-4 weeks post-viral infection. va ...19852989389
alterations in immunological reactivity in encephalomyocarditis virus-induced murine diabetes. i. defective primary igm plaque forming cell responses to sheep erythrocytes: correction by islet cell transplantation.increasing data suggest a possible viral aetiology of juvenile onset, insulin-dependent diabetes mellitus. the m variant of the encephalomyocarditis (emc) virus infects murine pancreatic beta cells and causes a diabetes like syndrome in susceptible strains of mice. abnormalities in immunological function have been documented in patients with diabetes mellitus and in spontaneous, streptozotocin-induced and alloxan-induced diabetes in animals. the present study documents a significant impairment o ...19852988831
a practical cytopathic effect/dye-uptake interferon assay for routine use in the clinical laboratory.the clinical value of interferon (ifn) level determinations has been demonstrated, but a practical assay procedure for routine use in the diagnostic laboratory has not been available. the authors examined the susceptibility of five cell lines (wish, hep-2, vero, a549, and wi-26va4) to vesicular stomatitis virus (vsv) and encephalomyocarditis virus (emcv), and compared the response of these cells to the inhibitory activity of ifn-alpha and ifn-gamma for the respective viral cytopathic effect (cpe ...19852988328
is insulin-dependent diabetes mellitus caused by coxsackievirus b infection? a review of the epidemiologic evidence.the evidence that coxsackievirus b plays a causal role in the etiology of insulin-dependent diabetes mellitus (iddm) is reviewed. this hypothesis is biologically plausible; one variant of another picornavirus, encephalomyocarditis virus, causes diabetes in genetically susceptible mice, but prior infection with another, serologically indistinguishable variant prevents this. the seasonal distributions of infection due to coxsackievirus b and of iddm are similar. case reports document coxsackieviru ...19852988099
immunologic behavior of lymphocytes in experimental viral myocarditis: significance of t lymphocytes in the severity of myocarditis and silent myocarditis in balb/c-nu/nu mice.to clarify the immune mechanism in myocarditis, we examined by immunofluorescence techniques the serial changes in percentages of t and b lymphocytes in the heart, spleen, and peripheral blood of dba/2 mice inoculated with encephalomyocarditis (emc) virus (experiment i). b cells were demonstrated by staining with fluorescein isothiocyanate (fitc)-labeled rabbit antimouse immunoglobulin (ig). t cells were demonstrated with rat anti-thy1.2 monoclonal antibody plus fitc-labeled antimouse ig. there ...19852986878
[natural interferon inducers: double-stranded rna of killer plasmids of saccharomyces cerevisiae].methods for isolation and purification of yeast double spiral rna (dsrna) are described. the most simple method includes reprecipitation of dsrna in solutions of lithium chloride and its interphase distribution in the system of chloroform: isoamilic alcohol: water. preparations with the content (w/w) of dsrna from 30 to 90 per cent were obtained. they possess a high interferon-inducing activity and an antiviral effect on the experimental infection caused by the virus of mouse encephalomyocarditi ...19852986543
shutoff of host translation by encephalomyocarditis virus infection does not involve cleavage of the eucaryotic initiation factor 4f polypeptide that accompanies poliovirus infection.studies were conducted to determine whether encephalomyocarditis virus infection causes proteolytic cleavage of any of the polypeptides which comprise eucaryotic initiation factor 4f. since no such alterations in the components of the initiation factor were detected, these observations confirmed that the mechanisms whereby encephalomyocarditis virus and poliovirus shut off host translation are different.19852985829
biochemical changes induced by coxsackie b4 virus in short-term culture of mouse pancreatic islets.isolated mouse pancreatic islets were infected in vitro with two strains of coxsackie b4 virus--a tissue culture-adapted strain and a mouse pancreas-adapted strain. within 48 h of infection changes had occurred in the biochemical activities of islets infected with the mouse pancreas-adapted strain of virus. basal insulin release was increased two-fold in these islets, while glucose-induced insulin secretion remained unchanged. insulin biosynthesis was greatly reduced at a stimulatory concentrati ...19852985140
sequence analysis of hepatitis a virus cdna coding for capsid proteins and rna polymerase.we report here the nucleotide sequence corresponding to two large regions of the hepatitis a virus (hav) genome. these comprise a sequence of 3274 bases corresponding to the 5' end of the genome, which includes the putative capsid protein region of this picornavirus, and 1590 bases corresponding to the 3' end of the genome, terminating in a 15-base poly(a) tract. these sequences revealed that hav had the characteristic genomic organization of picornaviruses: an open reading frame beginning appro ...19852984684
encephalomyocarditis virus and diabetes mellitus: studies on virus mutants in susceptible and non-susceptible mice.the so-called m-variant (especially subtype d) of encephalomyocarditis virus (emcv) induces a diabetes-like syndrome in certain mouse strains which may serve as a model of insulin-dependent diabetes mellitus (iddm) in man. the development and course of diabetes was influenced by a number of virus and host factors, among these being virus strain, virus dose, mouse strain, age, sex, and the host's immunological status. in a d-variant stock of emcv, we found a virus plaque variant (pv 2) diabetogen ...19852984315
clinical findings in acute viral myocarditis. with special attention to experimental and immunological evidence.according to analysis of 40,000 autopsies, the incidence of myocarditis is approximately 3.5%, the diagnosis of which had not been established in the majority prior to death. the clinical characteristics of myocarditis encompass a wide spectrum from an asymptomatic course to sudden cardiac death. symptoms and findings of congestive heart failure or an inflammatory virus disease may be present. there is usually evidence of nonspecific clinical, radiologic, electrocardiographic, echocardiographic ...19852984098
treatment of viral myocarditis with ribavirin in an animal preparation.the therapeutic effects of ribavirin, a broad-spectrum, antiviral agent, on experimental myocarditis caused by encephalomyocarditis (emc) virus were investigated. four-week-old dba/2 mice were inoculated with 10 plaque-forming units (pfu) of emc virus. ribavirin in a dose of 100 (group 1, n = 20), 200 (group 2, n = 10), or 400 mg/kg/day (group 3, n = 10) was administered subcutaneously on days 0 to 12 after virus inoculation, and animals were observed for 12 days. control animals were injected w ...19852982522
the reversion of temperature-sensitive mutants of encephalomyocarditis virus upon serial passage in three continuous cell lines.several temperature-sensitive mutants of encephalomyocarditis virus were serially passed numerous times in hela, l929 and bhk-21 cells. the reversion of most of the mutants occurred most rapidly in bhk-21 cells and least rapidly in hela cells.19852981972
respective role of each of the purine n-6 amino groups of 5'-o-triphosphoryladenylyl(2'----5')adenylyl(2----5')adenosine in binding to and activation of rnase l.we have synthesized a series of 2-5a (ppp5'-a2'p5'a2'p5'a) analogs in which each adenosine residue has been sequentially replaced by inosine: viz., ppp5'i2'p5'a2'p5'a, ppp5'a2'p5'i2'p5'a, and ppp5'a2'p5'a2'p5'i. these transformations enabled us to delineate the role of each of the three purine n-6 amino groups of 2-5a in determining oligonucleotide binding to and activation of the 2-5a-dependent endoribonuclease, rnase l. with the rnase l activity of both mouse l cells and human daudi lymphoblas ...19852981855
the effect of serum deprivation on the initiation of protein synthesis in mouse neuroblastoma cells.growth of mouse neuroblastoma cells becomes stationary when cultured in serum-free medium. within 60 h, the protein-synthesizing capacity of the cells declines to 25% as compared to that of exponentially growing cells. the transitional activity of the crude ribosomal salt washes from serum-deprived and control cells was compared in in vitro protein-synthesizing ph 5 systems. it appears that the ribosomal salt wash from serum-deprived cells has significantly (70%) lost its ability to support the ...19852981551
the mouse immune interferon receptor gene is located on chromosome 10.when mouse l cells are incubated with 32p-labeled recombinant murine immune interferon ( [32p]mu-ifn-gamma) and subsequently cross-linked with disuccinimidyl suberate, a major complex with an apparent molecular mass of 95,000-125,000 daltons can be visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. the complex was not formed when the binding was performed in the presence of excess unlabeled mu-ifn-gamma or when chinese hamster ovary cells were used. this complex therefore r ...19872952646
tuftsin a natural peptide with antiviral activity--structural basis of its action.the influence of tuftsin--the natural phagocytosis stimulating tetrapeptide and its elongated analogues--on the mortality of encephalomyocarditis virus-infected nmri mice was investigated. it seems that amino acids elongation in the parent peptide chain does not result in significant increase of antiviral activity of the compounds tested.19882897777
immunologic identification of lymphocyte subsets in experimental murine myocarditis with encephalomyocarditis virus. different kinetics of lymphocyte subsets between the heart and the peripheral blood, and significance of thy 1.2+ (pan t) and lyt 1+, 23+ (immature t) subsets in the development of myocarditis.to clarify the immune mechanism in myocarditis, immunofluorescence techniques with laser flow cytometry were used to examine serial changes in lymphocyte subsets in the heart, spleen, and peripheral blood of dba/2 and balb/c mice inoculated with encephalomyocarditis virus (experiment i). b cells were identified by staining with fluorescein isothiocyanate-labelled rabbit anti-mouse immunoglobulin. t-cell subsets were identified with rat anti-thy 1.2, and nonpolymorphic lyt 1 and lyt 2 monoclonal ...19872889539
immunomodulation by bordetella pertussis: antiviral effects.treatment of mice by intraperitoneal inoculation of pertussis vaccine or lipopolysaccharide extracted from b. pertussis will effect resistance to rabies virus, encephalomyocarditis virus, semliki forest virus, and herpes simplex virus. our previous observations indicated that treatment of c3h/hen (+/nu) and bdf1 mice with pertussis vaccine injected i.p. five days prior to a mouse adenovirus lethal dose i.p. challenge elicited resistance to clinical disease and death. susceptibility returned to a ...19852872109
induction of beta 2-interferon by tumor necrosis factor: a homeostatic mechanism in the control of cell proliferation.earlier studies showed that tumor necrosis factor (tnf) exerts a mitogenic effect in human diploid fibroblasts. here we demonstrate that purified e. coli-derived recombinant human tnf inhibits encephalomyocarditis virus replication in "aged" human fibroblasts. addition of neutralizing antibodies to human beta interferon (ifn-beta) blocked the antiviral action of tnf, indicating that this action is mediated by the generation of ifn-beta. we also show that antiserum to ifn-beta enhanced the mitoge ...19862871942
glucocorticoid-mediated establishment of an antiviral state coincident with other glucocorticoid-induced biochemical activities in l929 cells.incubation of l929 cells with three different glucocorticoids, hydrocortisone, dexamethasone and triamcinolone acetonide, rendered the cells unable to support plaque formation by several unrelated dna and rna viruses. the establishment of this antiviral state by dexamethasone coincided with an inhibition of cell growth and induction of glutamine synthetase activity. these steroid-mediated activities occurred only in cultures of l929 cells and not in cultures of rabbit skin or rat glioma cells.19852864388
derivatives of benzo(c)fluorene. vi. antiviral and interferon-inducing activities of three benzo(c)fluorenone derivatives in mice.the antiviral effect of 3 derivatives of benzo(c)fluorenone against encephalomyocarditis (emc) and vaccinia viruses in mice was compared with that of tilorone hydrochloride (thcl). all 3 derivatives were effective against either virus following single-dose oral application as well as after 4-times repeated subcutaneous (s.c.) treatment, but following oral administration only the vufb 14162 derivative exerted an antiviral effect corresponding to that of thcl. after s.c. application, however, vufb ...19852859757
structure/function studies on recombinant human gamma interferon.structure/function relationships for human gamma interferon (ifn-gamma) were investigated using recombinant dna-derived homologues produced in e. coli. the various biological effects examined were antiviral, growth inhibitory and 2-5a synthetase activities, as well as receptor binding characteristics. specific structural changes led to ifn-gamma homologues with defined alterations in biological activities. amino acid residue changes at the hydrophobic core of the molecule resulted in two homolog ...19882855574
[viral infections in medicine. 4. viral myocarditis and cardiomyopathy]. 19882854544
application of immunoassay of encephalomyocarditis virus in cell culture with enzyme-labeled virus-specific monoclonal antibodies for rapid detection of virus, neutralizing antibodies, and interferon.encephalomyocarditis virus (emcv)-specific monoclonal antibody um 21.1 labeled with horseradish peroxidase was used to detect emcv in l-cell monolayers. this direct enzyme immunoassay of emcv, performed in wells of 96-well plates, could be applied for various purposes, such as early detection of virus multiplication, determination of 50% tissue culture infective doses, and rapid titration of interferon and emcv-neutralizing antibodies. multiplication of emcv is indicated by a rapid increase of t ...19882852674
encephalomyocarditis virus infection in pigs. 19882850788
[experimental viral-bacterial infections. ii. development and course of mixed emc virus and staphylococcus aureus infection in mice]. 19882850413
effects of cyclosporine on encephalomyocarditis virus infection in mice. 19882848238
[an enzyme hydrolyzing the covalent bond between rna and vpg of picornaviruses. substrates and methods of analysis of activity].some new substrates--rna-vpg, rna-peptide and a small fragment of rna peptide--labelled at the peptide component with [125i]bolton-hunter reagent have been proposed for use in the isolation and characterization of the enzyme hydrolyzing the phosphodiester bond between the vpg protein and encephalomyocarditis virus rna. a novel procedure for the analysis of the specific enzyme activity is based on thin-layer chromatography of hydrolytic products on silicagel or polyethylenimine cellulose. the mol ...19882847820
mode of action of a new type of udp-glucose analog against herpesvirus replication.the mode of action of a new type of udp-glucose analog against herpes simplex virus type 1 (hsv-1) replication was examined. the analog showed good selectivity and potent activity. at 10 micrograms/ml, p-536 inhibited the formation of infectious hsv-1 by more than 90%, whereas at 100 micrograms/ml it had no cytotoxic effects, as evidenced by phase-contrast microscopy. p-536 showed a wide spectrum of action and was active against hsv-1, adenovirus type 5, vaccinia virus, poliovirus type 1, enceph ...19882847650
the natural history of encephalomyocarditis virus-induced myositis and myocarditis in mice. viral persistence demonstrated by in situ hybridization.picornaviruses can initiate chronic inflammation that persists after the virus can no longer be cultured from inflamed tissues. in an attempt to understand this transition we have sought evidence for viral persistence by methods that detect viral genome independent of whether or not whole competent virus is present. in mice infected with a myotropic variant of encephalomyocarditis virus, emc-221a, virus can be cultured in high yield at 1 wk and in low yield at 2 wk from skeletal muscle, heart, a ...19882846742
reduced tumorigenicity of rodent tumour cells and tumour explants following infection with wild type and mutant herpes simplex virus, bovine mammillitis virus and encephalomyocarditis virus.the tumorigenicity of neoplastic hamster and mouse cell lines and tumour explants was reduced by infection with herpes simplex virus (hsv-1), a thymidine-kinaseless mutant of herpes simplex virus, namely 'mdk', encephalomyocarditis virus (emc) and bovine mammillitis virus (bmv). there was an approximate relationship between duration of virus infection in vitro and reduction in incidence and/or rate of tumour development. the rate of tumour development was also reduced by 'site inoculation' of vi ...19882846027
comparison of the nucleotide sequences of diabetogenic and nondiabetogenic encephalomyocarditis virus.the nucleotide sequences of a diabetogenic variant of encephalomyocarditis (emc) virus (d variant, ifp- phenotype) and a nondiabetogenic variant of emc virus (b variant, ifp+ phenotype) have been compared. these variants differ at eight sites. the poly(c) tract of emc-b is three bases shorter than that of emc-d. of the seven sites at which nucleotide substitutions were confirmed using rna templates, four resulted in amino acid changes in three different proteins, the leader peptide, 1b (vp2), an ...19882845665
relationship of p220 cleavage during picornavirus infection to 2a proteinase sequencing.infection of hela cells by poliovirus results in an abrupt inhibition of host cell protein synthesis. it is thought that the mechanism of this inhibition involves proteolytic cleavage of the p220 component of the cap-binding protein complex, thereby causing functional inactivation of the cap-binding protein complex and preventing capped (cellular) mrnas from binding ribosomes. current data suggest that the viral proteinase 2a indirectly induces p220 cleavage via alteration or activation of a sec ...19882845133
studies on the role of the 2'-5'-oligoadenylate synthetase-rnase l pathway in beta interferon-mediated inhibition of encephalomyocarditis virus replication.interferons inhibit the replication of vesicular stomatitis virus (vsv), but not of encephalomyocarditis virus (emcv), in mouse jlsv-11 cells. we report the isolation of clonal derivatives from this cell line in which the replication of both viruses is impaired by interferons. these clones were selected from the parental line by virtue of their rescue by interferon treatment from the cytopathic effects of emcv infection. in one such clone, rk8, the replication of vsv and emcv and the production ...19882841470
a segment of the 5' nontranslated region of encephalomyocarditis virus rna directs internal entry of ribosomes during in vitro translation.picornavirus rnas are uncapped messengers and have unusually long 5' nontranslated regions (5'ntrs) which contain many noninitiating aug triplets. the translational efficiency of different picornavirus rnas varies between different cell-free extracts and even in the same extract, such as micrococcal nuclease-treated rabbit reticulocyte lysates. the effect of the poliovirus 5'ntr on in vitro translation was compared with that of the 5'ntr of encephalomyocarditis virus by the use of synthetic mrna ...19882839690
histopathology of balb/c mice infected with the d variant of encephalomyocarditis virus.balb/c mice infected with 10(5) pfu of the d variant of encephalomyocarditis virus were examined histopathologically during the subacute stage of infection. main pathologic changes were observed in the brain, heart (massive myocardial necrosis with subsequent organization (i.e., replacement of necrotic myocardium by connective tissue) and congestion and dilatation of the right ventricle) and pancreas (moderate degranulation of beta-cells resulting in elevation of blood glucose level). the brain ...19882839220
detection of antibody to encephalomyocarditis virus in mummified or stillborn pigs.fetal sera or thoracic fluids of the abnormal fetuses from different swine farms were tested for the presence of antibody to encephalomyocarditis (emc) virus. sera of swine fetuses infected experimentally with emc virus were also tested. high antibody titers ranging 1:128-4096 were detected in both field and experimental samples. detection of the positive antibody was associated with clinical herd history of increased mummification, stillbirth and neonatal death. the results suggest that emc vir ...19882839129
quinazolyl benzophenothiazines as potential antiviral agents.the reaction of anthranilic acid with excess of benzoylchloride in the presence of pyridine yielded 2-phenyl(3h) 4-oxo-3,1-benzoxazine(i). interaction of (i) with 2-aminoethanol afforded 2-phenyl-3-hydroxy-ethyl(3h) 4-oxo-quinazoline (ii). the condensation of (ii) with beta-naphthol in the presence of conc. h2so4 gave 1-(2'-phenyl-3'-ethyl-4'-oxo-quinazolyl) beta-naphthol (iii). the reaction of (iii) with primary aromatic amines in the presence of anhydrous zinc-chloride afforded 1-(2'-phenyl-3' ...19882837789
synthesis, antiviral activity, and conformational characterization of mouse-human alpha-interferon hybrids.reciprocal hybrids were constructed between human and mouse interferons (ifns), and their antiviral activity was examined on different target cells and compared to the activity of the parental molecules. in addition, we used a number of predictive algorithms on a data base of the available alpha-interferon sequences to propose a working model for the overall conformation of the alpha-interferon molecule that is consistent with the structural predictions. remarkable conservation within the predic ...19882837469
identification of antigenic differences between the diabetogenic and non-diabetogenic variants of encephalomyocarditis virus using monoclonal antibodies.the m variant of encephalomyocarditis (emc) virus consists of two biologically distinct variants: one, diabetogenic d variant (emc-d) and the other, non-diabetogenic b variant (emc-b). these two variants cannot be distinguished by hyperimmune sera. monoclonal antibodies were generated against emc-d or emc-b to identify antigenic differences between these two variants. fourteen independent hybrid cell lines, selected from seven separate fusions of mouse myeloma cells to spleen cells isolated from ...19882836552
[an inhibitor of translation in cellular mrna containing the 100s structure from krebs ii ascites carcinoma cells infected with encephalomyocarditis virus].as shown previously, the bulk of cellular mrna in krebs ii ascite carcinoma cells infected with encephalomyocarditis virus during active virus-specific synthesis is bound to ribosomes within the 100s structure which is inactive in protein synthesis. in order to elucidate the reasons for the translation repression of cellular mrna within the 100s structure, a fraction of loosely bound proteins which are liberated by treatment of the 100s structure with 0.5 m kcl an which contain sum translation f ...19882833939
[4,5-s-rnai in virus-specific polyribosomes from ascitic krebs 2 carcinoma cells infected with encephalomyocarditis virus].polyribosomes of krebs 2 ascite carcinoma cells non-infected and infected with encephalomyocarditis (emc) virus contain a heterogeneous population of low molecular weight small rnas. analysis of the rnas by polyacrylamide gel electrophoresis did not reveal any qualitative differences in the small rna sets within the composition of polyribosomes from virus-infected and non-infected cells. however, the content of one of the small rnas was markedly elevated in polyribosomes from virus-infected cell ...19872833938
amino acid differences in capsid protein, vp1, between diabetogenic and nondiabetogenic variants of encephalomyocarditis virus.the genes for the major capsid protein, vp1(1d), of both diabetogenic d variant (emc-d) and nondiabetogenic b variant (emc-b) of encephalomyocarditis virus were cloned by using two synthetic primers which are common to both emc-d and emc-b. the cloned genes were mapped for major restriction enzyme sites including acci, bamhi, ecori, hincii, kpni, pvuii, ssti, taqi, and xbai. among those nine restriction enzyme sites, only the taqi site distinguished emc-d genome from the counterpart of emc-b gen ...19882833013
virus vaccine and passive immunization for the prevention of viral myocarditis in mice.the effect of an inactivated virus vaccine and passive immunization was studied in an animal model of viral myocarditis. encephalomyocarditis (emc) virus was inactivated by 1:4000 formalin. four-week-old cd-1 mice were injected subcutaneously and after 7 days intravenously with 0.1 ml vaccine. seven days after the second vaccine, live emc virus was inoculated intraperitoneally. none of ten mice with vaccine treatment died or showed myocardial lesions after challenge with emc virus, whereas all t ...19872832629
asbestos-mediated transfection of mammalian cell cultures.the capacity of asbestos to mediate transfection was tested in a rapid and relatively simple system: picornavirus rnas and mammalian cells in vitro. thirteen asbestos samples, including amosite, anthophyllite, chrysotile, and crocidolite, 4 picornaviruses (poliovirus 1 and 2, echovirus 7, and encephalomyocarditis virus), and 4 cell lines (cli, chimpanzee liver; kb, human carcinoma; eta, monkey kidney; nih 3t3, mouse embryo) were tested. the results showed that all asbestos samples mediated trans ...19882832359
sequence homology between encephalomyocarditis virus protein vpi and histidyl-trna synthetase supports a hypothesis of molecular mimicry in polymyositis.the amino acid sequence of histidyl-trna synthetase, the jo-i antigen of polymyositis, has been compared with that of the polyprotein of encephalomyocarditis virus (emcv), an agent shown recently to produce polymyositis symptoms in mice. it is shown that there is significant sequence homology between a region of the synthetase formerly identified by us as a possible epitope and a region on the coat protein vpi of emcv. the viral protein also shows significant sequence homology with several muscl ...19882830474
prophylaxis and therapy of virulent encephalomyocarditis virus infection in mice by monoclonal antibodies. brief report.two encephalomyocarditis virus (emcv) neutralizing monoclonal antibodies (mas), recognizing different determinants on emcv, were both able to protect mice prophylactically and therapeutically against a lethal dose of emcv.19882829795
inactivation of 12 viruses by heating steps applied during manufacture of a hepatitis b vaccine.the efficacy of two heating cycles (90 sec at 103 degrees c and 10 hr at 65 degrees c) used during manufacture of a plasma-derived hepatitis-b vaccine was validated for the inactivation of 12 virus families. a period of 15 min warming up to 65 degrees c had already completely inactivated representatives of nine virus families, ie, poxvirus (vaccinia), picornavirus (encephalomyocarditis virus), togavirus (sindbis virus), coronavirus (mouse hepatitis virus), orthomyxovirus (influenza virus), rhabd ...19872828525
selective inhibition of proteins synthesized from different mrna species in reticulocyte lysates containing l-pyrroline-5-carboxylic acid.l-pyrroline-5-carboxylic acid is a naturally occurring nonprotein amino acid present in human plasma that changes concentrations with diet. l-pyrroline-5-carboxylic acid inhibited net synthesis of globin in untreated reticulocyte lysates in a dose dependent manner. this inhibition was greater than that observed with equimolar gssg or nadp+ and was prevented by a nadph generating system. l-pyrroline-5-carboxylic acid also inhibited net synthesis of proteins from brome mosaic and alfalfa mosaic vi ...19882827672
an apparent deletion of an oligonucleotide detected by rna fingerprint in the nondiabetogenic b variant of encephalomyocarditis virus is caused by a point mutation.the diabetogenic d variant of encephalomyocarditis virus (emc-d) was previously shown to be different from the nondiabetogenic b variant of encephalomyocarditis virus (emc-b) by a single spot in an oligonucleotide fingerprint after rnase t1 digestion of their genomic rnas. an oligoribonucleotide was missing from emc-b but was present in emc-d. the oligoribonucleotide specific to emc-d was isolated from a two-dimensional polyacrylamide gel and sequenced as 5'-acaaucucacuuuuccaacaacag-3'. molecula ...19882826821
on the interrelationship between virus concentration and pathological changes of pancreas in mice induced by emc-virus. 19872826531
encephalomyocarditis virus-induced diabetes mellitus in mice: model of viral pathogenesis.encephalomyocarditis (emc) virus induction of diabetes mellitus in mice has proven to be an excellent experimental model for the pathogenesis of viral disease. in sjl and dba/2 mice, diabetes results exclusively from the infection and damage of beta cells by the virus. in addition, in balb/cby mice subclinical beta cell damage caused by the virus is followed by autoimmune beta cell destruction, which results in hyperglycemia. studies of two closely related plaque variants (emc-d and emc-b) selec ...19872825321
site-specific mutations at a picornavirus vp3/vp1 cleavage site disrupt in vitro processing and assembly of capsid precursors.most proteolytic cleavages within the picornavirus polyproteins are carried out by viral protease 3c. for encephalomyocarditis virus, the protease 3c-catalyzed processing occurs between gln-gly or gln-ser amino acid pairs which are flanked by proline residues, but the sequence-specific constraints on recognition and cleavage by the enzyme are not completely understood. to examine alternative cleavage site sequences, we constructed a cdna plasmid which expresses the viral l-p1-2a capsid precursor ...19872824807
factors affecting the infection of the d variant of encephalomyocarditis virus in the b cells of c57bl/6j mice.the d variant of encephalomyocarditis virus is capable of infecting most inbred strains of mice. however, only certain strains are susceptible to the diabetogenic effect of this virus. in order to understand why some inbred strains do not become diabetic, the pathogenesis of infection was studied in diabetes-resistant c57bl/6j mice. it was the purpose of the investigation to ascertain whether specific host defense factors might play a crucial role in the mechanism of resistance. to determine whe ...19872824265
impaired cytokine response in male icr swiss mice after infection with d variant of encephalomyocarditis virus.the basis for the resistance of the female and the susceptibility of the male icr swiss mouse to the diabetogenicity of the d variant of encephalomyocarditis virus (emcv-d) is unknown. this pattern of disease resistance and susceptibility can be reversed if females are treated with testosterone and males are treated with estrogen before virus infection. as a possible explanation for this sex difference in disease development, differences in early antiviral host responses were explored. cellular ...19872824259
possible mechanisms in the pathogenesis of virus-induced diabetes mellitus.insulin-dependent diabetes mellitus results from destruction of pancreatic beta cells. viruses and autoimmunity have been implicated as possible causes of beta cell destruction in genetically predisposed individuals. the evidence for viruses comes largely from experiments in animals, but several studies in humans point to viruses as triggers in the pathogenesis of diabetes in some cases. in animal models, at least 4 different possible mechanisms for virus-induced diabetes have been proposed. the ...19872824113
pathogenesis and preventive and therapeutic trials in an animal model of dilated cardiomyopathy induced by a virus.an animal model of dilated cardiomyopathy following encephalomyocarditis (emc) virus has been developed. virus was isolated from mouse hearts and viral antigens were detected in the myocardium until the second week of infection, but neither was found thereafter. differences were found among different strains of mice in the frequency of occurrence and severity of myocarditis, and even in the character of the pathologic lesions. thus, genetic factors may play an important role in the pathogenesis. ...19872822976
prodromal immune manifestations in emc-m virus induced diabetes: islet bound and circulating antibodies, and changes in lymphocyte subsets.the thymus-dependence of the encephalomyocarditis (emc-m) virus induced diabetes has been demonstrated in comparative studies of normal and immunodeficient mice. since the lymphocytic infiltration in the islets of langerhans is modest during the virus infection, we have looked for possible indications of humoral immune mechanisms. using fluorescence microscopy the presence of immunoglobulins in the islets could be shown 3 days after emc-m-virus inoculation, gradually disappearing about day 14. t ...19872820645
enhanced in vivo therapeutic response to interferon in mice with an in vitro interferon-resistant b-cell lymphoma.a stable subline of 38c13 b-cell lymphoma (sir-1) resistant to the antiproliferative effects of alpha-interferon (ifn) was isolated. in addition to defects in antiproliferative effects of ifn, sir-1 is defective in ifn-mediated antiviral activity against both encephalomyocarditis virus and vesicularstomatitis virus. it is also defective in the induction of 2'-5'-oligoadenylate synthetase mrna and enzyme activity, enhancement of h-2 antigen expression, and transient induction and subsequent repre ...19892787203
characteristics of lymphocytes cultured from murine viral myocarditis specimens: a preliminary and technical report.long-term culture of lymphocytes from myocardial biopsy specimens has been reported. to test the usefulness of this technique in experimental models of murine myocarditis, the culture and characterization of lymphocytes from mice infected with encephalomyocarditis virus or coxsackievirus b3 were studied. lymphocytes were successfully cultured from three hearts of encephalomyocarditis virus-infected mice in interleukin 2-containing culture. after approximately 10 days, lymphocytes migrated out of ...19892768726
rapid development of renal lesions in diabetic dba mice infected with the d-variant of encephalomyocarditis virus (emc-d).marked hyperglycaemia and renal lesions developed rapidly in dba mice infected with 10 plaque-forming units of the d-variant of encephalomyocarditis virus (emc-d). renal alterations were demonstrated in the glomeruli, tubular epithelium and small vessels 2 months after infection. glomerular changes were characterized by mesangial thickening due to an increase of basement membrane-like material in the mesangial matrix. nodular glomerular lesions were commonly observed 3 months after infection, wh ...19892765387
double-stranded rna-dependent protein kinase and 2-5a system are both activated in interferon-treated, encephalomyocarditis virus-infected hela cells.activation of the interferon-inducible, double-stranded rna-dependent protein kinase was monitored in monolayer cultures of control and interferon-treated hela cells infected with encephalomyocarditis virus. the extent of phosphorylation in the intact cell of the alpha-subunit of eucaryotic protein synthesis initiation factor eif2 by the kinase was determined for the first time in this type of system, using a two-dimensional immunoblot technique. virus protein synthesis and the kinetics of activ ...19852582150
[effect of thymosin on interferon production and antiviral resistance in mice].the 5th and 6th fractions of thymosin, a hormone of the thymus gland, stimulated interferon production both in vivo (experiments in white and cba mice) and in vitro in cba mouse splenocytes when different interferon inducers were used (phage dsrna, poly(g): poly(c), ndv, and mitogens). the highest stimulating effect in vivo was observed with interferon induction 6-8 hours after thymosin administration. an increase in production of both alpha/beta and gamma interferons under the influence of thym ...19852581368
role of sex and early interferon production in the susceptibility of mice to encephalomyocarditis virus.adult female swiss mice showed a greater resistance to intraperitoneal (i.p.) infection with encephalomyocarditis virus (emcv) than male mice. this difference was not observed in weanling mice, in castrated adult mice or in adult mice injected intracerebrally. administration of antibody to mouse interferon alpha/beta enhanced the virulence of emcv for both sexes and no difference was then observed in susceptibility between male and female mice. six h after emcv infection, serum interferon titres ...19852580047
the preparation of antiidiotypic antibodies against human beta-interferon. 19892576603
serological survey for viral diseases in the cayo santiago rhesus macaque population.the free-ranging population of rhesus monkeys (macaca mulatta) on cayo santiago was sero-surveyed for human measles, simian virus 40, b virus (herpes simiae), rhesus cytomegalovirus, human and simian retroviruses and encephalomyocarditis virus to determine the prevalence of these viruses in the colony. the results of this study indicate that the colony is free of sv40, htlviii (hiv-1), stlviii (siv) and srv1; has a low prevalence of measles and emcv; and high prevalence rates for b virus, cmv an ...19892571176
serologic, virologic, and histopathologic observations of encephalomyocarditis virus infection in mummified and stillborn pigs.stillborn and mummified swine fetuses from swine farms experiencing reproductive problems were investigated for evidence of infection with encephalomyocarditis (emc) virus by fetal serology, virus isolation, and histopathologic examination. fetal sera or thoracic fluids of 478 abnormal fetuses collected during january through december 1987 were tested for the presence of antibody specific to emc virus. of 478 samples tested, 175 (36.6%) had emc virus serum neutralizing antibody titers of 1:64 or ...19892562190
sex-dependent, early cytokine production by nk-like spleen cells following infection with the d variant of encephalomyocarditis virus (emcv-d).spleen cell cultures from diabetes-resistant icr swiss females exhibited an increase in expression of ia antigens 24 hours post-infection (pi) with emcv-d while comparable spleen cell cultures from diabetes-susceptible males of this strain did not exhibit this increase in ia antigens expression. a monoclonal antibody specific for mouse interferon-gamma (ifn gamma) eliminated this increase in ia antigens expression. interferon-gamma (ifn gamma) and interleukin 2 (il-2) production by emcv-d-infect ...19892560916
pathogenic properties of encephalomyocarditis virus isolates in swine fetuses.the pathogenicity of 5 different encephalomyocarditis (emc) virus isolates was investigated in swine fetuses following injection of each virus in utero. laparotomies were performed on 3 pregnant sows in early mid-third (39-40 days) of gestation and on 5 sows in the late mid-third (70-72 days) of gestation, and groups of fetuses were inoculated with different viruses into the amniotic sacs. the uninoculated fetuses served as controls. thirty-five (71.4%) of 49 infected and 1 of 26 control fetuses ...19892558630
inducible expression of encephalomyocarditis virus 3c protease activity in stably transformed mouse cell lines.an inducible expression vector system has been developed to facilitate the study of the effects of individual virus gene products on cell function. the vector utilizes the mouse metallothionein promoter carried on the bovine papillomavirus genome. conditions which optimize the induced expression of open reading frames inserted downstream from the mouse metallothionein promoter have recently been described. in this communication we describe the use of this system to clone and express the encephal ...19892555538
tropism and histopathology of the d, b, k, and mm variants of encephalomyocarditis virus.variants of encephalomyocarditis virus (emcv), which are immunologically indistinguishable by hyperimmune serum, produce different disease syndromes in mice. for instance, in icr swiss male mice, emcv-b produces no overt illness, emcv-mm produces severe neurological signs followed by death, emcv-d destroys pancreatic beta cells producing a disease syndrome resembling insulin-dependent diabetes mellitus, and emcv-k is lethal but produces no overt signs of infection. the present study was done to ...19892555446
hydrolysis of a series of synthetic peptide substrates by the human rhinovirus 14 3c proteinase, cloned and expressed in escherichia coli.the 3c proteins of several picornaviruses, including poliovirus, foot-and-mouth disease virus (fmdv) and encephalomyocarditis virus (emcv), have been demonstrated to be cysteine-type proteinases, involved in the processing of the respective polyproteins expressed by the monocistronic rna genome. nucleotide sequencing data have indicated that the human rhinovirus 14 (hrv-14) rna genome encodes a homologous 3c protein. the hrv-14 3c protein was purified to homogeneity from escherichia coli express ...19892555433
anti-interferon antibodies to interferon-alpha 2b: results of comparative assays and clinical perspective.previous studies have reported a low (less than 3%) incidence of anti-interferon (ifn) serum neutralizing antibodies following treatment with ifn-alpha 2b. since this result contrasts with a higher incidence reported with ifn-alpha 2a, the question has been raised whether differences in assay techniques and patient comparability rather than inherent differences in the molecules might account for the reported differences in antibody incidence. in this report two patient groups, 151 hairy cell leu ...19892553829
susceptibility of four species of small rodents to encephalomyocarditis (emc) virus infection.the d variant of encephalomyocarditis virus (10(1)-10(5) pfu/head) was intraperitoneally inoculated into 4 species of small rodents, rats, mice, syrian hamsters, and mongolian gerbils, and the susceptibility of these animals to emc virus was examined virologically and histopathologically 3 days after infection. viral replication was detected in the brain (mice), in the heart (mice and gerbils), and in the pancreas (mice, hamsters, and gerbils). no viral replication was detected in rats. histopat ...19892553452
degradation of cellular proteins during poliovirus infection: studies by two-dimensional gel electrophoresis.picornaviruses encode for their own proteinases, which are responsible for the proteolytic processing of the polyprotein encoded in the viral genome to produce the mature viral polypeptides. the two poliovirus proteinases, known as proteins 2a and 3c, use the poliovirus-encoded polyprotein as a substrate. the possibility that these poliovirus proteinases also degrade cellular proteins remains largely unexplored. high-resolution two-dimensional gel electrophoresis indicates that a few cellular pr ...19892552149
identification and production of monoclonal antibodies against a discriminating protein molecule between b and d variants of encephalomyocarditis virus.the protein profiles of two emc virus variants b and d in the infected l929 fibroblast lysates were examined using one- and two-dimensional gel electrophoresis. both variants yielded a protein molecule of similar molecular weight but differing in its isoelectric point (pi). the b variant lysate yielded a molecule with pi congruent to 7.5 whereas the same molecule from the d lysate focused at pi congruent to 5.2. a monoclonal antibody (mca) produced against pi congruent to 5.2 fraction of the d v ...19892551748
treatment of encephalomyocarditis virus-induced central nervous system demyelination with monoclonal anti-t-cell antibodies.infection of balb/c mice with the m variant of encephalomyocarditis virus resulted in the development of a paralytic syndrome in 7 to 10 days. the paralysis was maximal during the period of viral clearance; most of the animals recovered from the initial deficit and showed no delayed recurrences. pathologically, the white matter of brain and spinal cord showed well-demarcated areas of perivascular cuffing, demyelination, and, during recovery, remyelination by oligodendrocytes--all suggestive of p ...19892550666
tumor necrosis factor acts synergistically with autocrine interferon-beta and increases interferon-beta mrna levels in human fibroblasts.medium of untreated human fs-4 foreskin fibroblasts contained a factor which, upon the addition of exogenous tumor necrosis factor (tnf), inhibited encephalomyocarditis virus replication when neither medium alone nor tnf alone were effective. this antiviral activity was abolished by a monoclonal antibody to human interferon (ifn)-beta, suggesting that the active component in the medium from untreated fs-4 cells was ifn-beta, present at subeffective concentrations. in addition, we show that untre ...19892550437
an unusual coordinated cleavage event in the processing of encephalomyocarditis virus polypeptides.during the translation of encephalomyocarditis virus rna in the rabbit reticulocyte lysate system, polypeptides 2b and 2c, encoded by the central region of the genome, appear simultaneously in the absence of any detectable 2bc precursor and at a time when translation has advanced into the region coding for polypeptide 3c. this implies the operation of closely coordinated proteolysis at two sites, with at least one cleaved by the virus-coded protease activity 3c.19892549719
regulation of macrophage growth and antiviral activity by interferon-gamma.interferons, in addition to their antiviral activity, induce a multiplicity of effects on different cell types. interferon (ifn)-gamma exerts a unique regulatory effect on cells of the mononuclear phagocyte lineage. to investigate whether the antiviral and antiproliferative effects of ifn-gamma in macrophages can be genetically dissociated, and whether ifn-alpha and ifn-gamma use the same cellular signals and/or effector mechanisms to achieve their biologic effects, we have derived a series of s ...19892549078
cap-independent translation of mrna conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage t7 hybrid expression system.a recombinant vaccinia virus that directs the synthesis of bacteriophage t7 rna polymerase provides the basis for the expression of genes that are regulated by t7 promoters in mammalian cells. the t7 transcripts, which account for as much as 30% of the total cytoplasmic rna at 24 hr after infection, are largely uncapped. to improve the translatability of the uncapped rna, the encephalomyocarditis virus (emcv) untranslated region (utr) was inserted between the t7 promoter and the chloramphenicol ...19892548200
conservation of the secondary structure elements of the 5'-untranslated region of cardio- and aphthovirus rnas.an analysis of published nucleotide sequences of the 5'-untranslated region (5'-utr) of 7 cardioviruses and 3 aphthoviruses has allowed us to derive a consensus secondary structure model that differs from that previously proposed for the 5'-utr of entero- and rhinoviruses, though all these viruses belong to the same family, picornaviridae. the theoretical model derived here was experimentally supported by investigating the accessibility of encephalomyocarditis virus rna to modifications with dim ...19892548167
[changes in the myocardium in encephalomyocarditis of pigs].the myocardial lesions of pigs (age from 2.5 to 3.5 month) with encephalomyocarditis virus infection are presented. the disease is caused by members of the genus cardiovirus (family picornaviridae). in greece it was first observed 1987 in two stables of pig-breeders with a mortality of 80%. the macroscopic changes are characterized by a myocardial degeneration with greyish pale linear or round foci and by an accumulation of a clear red to yellow fluid in the pericardium and the pleural cavity. t ...19892547556
human infection with encephalomyocarditis virus in new south wales. 19892547141
a serologic survey of encephalomyocarditis virus infection in pigs in italy.neutralizing antibodies to encephalomyocarditis virus (emcv) were found in 69.13% of the serum samples obtained from pigs from representative regions of italy. the antibodies are distributed fairly uniformly throughout the swine populations with all age groups being equally involved.19892546020
reduced activity of the interferon-induced double-stranded rna-dependent protein kinase during a heat shock stress.previous studies have shown that the antiviral response induced by interferon in murine cells could be degraded after a heat shock. here we have confirmed that a similar effect occurs also in interferon-treated human hela cells subjected to a heat shock. in addition, we have investigated the fate of the interferon-induced, double-stranded rna-dependent protein kinase in heat-shocked cells. this protein kinase is a mr 68,000 protein (p68 kinase) which, when autophosphorylated, catalyzes phosphory ...19892545700
di-octyl phthalate induced altered host resistance: viral and protozoal models in mice. 19892545652
viral myocarditis and cardiomyopathy. 19892544753
production and in vivo biologic actions of recombinant mouse interferon alpha 2.we have expressed a recombinant mouse interferon alpha (r.mu-ifn alpha 2) in escherichia coli under the control of a tryptophan promoter using a synthetic adaptor formed by annealing two partially complementary oligonucleotides which introduced an atg start codon and re-established the complete coding sequence of the mature ifn alpha 2 protein in the expression vector. levels of up to 10(7) reference units of mu-ifn alpha 2 per liter of culture were obtained using this construction. this recombi ...19892544081
genomic differences between the diabetogenic and nondiabetogenic variants of encephalomyocarditis virus.plaque purification of the m variant of encephalomyocarditis (emc-m) virus resulted in the isolation of two stable variants. one is a highly diabetogenic d variant (emc-d) and the other is a nondiabetogenic b variant (emc-b). the cdna of emc-d and emc-b genomes were cloned and seven overlapping cdna clones were selected to cover the entire genome except the 5'-end 310 bases which were determined by rna-dependent dna sequencing and enzymatic rna sequencing. each clone was restriction-mapped, subc ...19892541543
viral myocarditis: immunopathogenesis and the effect of immunosuppressive treatment in a murine model.serial changes in lymphocyte subset were investigated in an animal model of encephalomyocarditis (emc) virus myocarditis in mice. the lymphocyte subset in the peripheral blood was measured by flow cytometry and, t cell subset in the myocardium was stained by labeled avidin-biotin technique. helper-inducer t cells (lyt 1 cells) decreased significantly on day 7, but there was no change in cytotoxic-suppressor t cells in the peripheral blood. during the acute stage a significant number of t cells ( ...19892541264
cloning and synthesis of infectious cardiovirus rnas containing short, discrete poly(c) tracts.mengovirus rna transcripts with 5' noncoding poly(c) tracts of c8, c12, and c13uc10 have been synthesized in vitro from cdna clones and shown to be infectious to hela cells. a chimeric clone has also been constructed which links the 5' end from one mengovirus clone (299 nucleotides, containing c13uc10) to a 7,424-base fragment derived from the 3' end of encephalomyocarditis (emc) virus. progeny virus isolated after transfection with the clone-derived rnas had the same poly(c) tracts, mengovirus- ...19892538661
initiation of protein synthesis by internal entry of ribosomes into the 5' nontranslated region of encephalomyocarditis virus rna in vivo.expression vectors that yield mono-, di-, and tricistronic mrnas upon transfection of cos-1 cells were used to assess the influence of the 5' nontranslated regions (5'ntrs) on translation of reporter genes. a segment of the 5'ntr of encephalomyocarditis virus (emcv) allowed translation of an adjacent downstream reporter gene (cat) regardless of its position in the mrnas. a deletion in the emcv 5'ntr abolishes this effect. poliovirus infection completely inhibits translation of the first cistron ...19892538648
molecular identification of diabetogenic viral gene.the best evidence that viruses have a causative role in the pathogenesis of insulin-dependent diabetes mellitus comes from experiments in mice infected with encephalomyocarditis (emc) virus. when sjl/j male mice were inoculated with a highly diabetogenic emc-d virus, diabetes developed in 95% of the animals. in contrast, none of the mice inoculated with a nondiabetogenic emc-b virus became diabetic. tissue culture experiments showed that emc-b induces considerable amounts of interferon, whereas ...19892537245
proteolytic cleavage of encephalomyocarditis virus capsid region substrates by precursors to the 3c enzyme.picornavirus protease 3c is normally released from its p3 precursor by two successive self-cleavage reactions. the free enzyme can then catalyze most of the remaining processing events within the viral polyprotein. to investigate the role of the 3c precursors in the processing cascade, we constructed cdna clones which expressed genetically altered forms of the encephalomyocarditis p3 region in vitro. site-specific substitutions were introduced into the gln-gly residues at the 3b-3c and 3c-3d jun ...19892536819
myocardial uptake of antimyosin monoclonal antibody in a murine model of viral myocarditis.the myocardial uptake of 125i- and 131i-antimyosin monoclonal antibody fab in experimental myocarditis in balb/c mice induced by encephalomyocarditis virus was studied. the biodistribution of 125i-antimyosin demonstrated that the highest ratio of radioactivity appears in the heart of infected mice on day 14 (the ratio of percent dose per gram for the organ to percent dose per milliliter for blood; 9.75 +/- 2.79 vs. 1.27 +/- 0.78 at 24 hours in inoculated mice vs. control mice). there was no stat ...19892536600
experimental encephalomyocarditis virus infection in mongolian gerbils (meriones unguiculatus).two strains of mongolian gerbils (meriones unguiculatus), tumble brook (tum) and japan medical science (jms), were intraperitoneally inoculated with the d variant of encephalomyocarditis virus (emc-d) and killed 3 days later. mortality was significantly higher in females than in males. evidence of viral replication was detected in the heart of both strains and in the pancreas of the tum strain. histopathological alterations were found in the heart and pancreas. heart lesions involved foci of nec ...19892536510
different antiviral activity and cell specificity of interferon preparations produced by mouse peritoneal cells at 37 degrees c and at 26 degrees c.three sublines of mouse l cells and mouse embryo fibroblasts were used for determination of the antiviral activity of mouse interferons produced by nonadherent peritoneal exudate cells incubated either at 37 degrees c or at 26 degrees c. ifn produced at 37 degrees c or at 26 degrees c had the same antiviral activity in l borgen, l929 cells. however, in mec ifn-37 degrees had relatively higher activity than ifn-26 degrees. of the interferon investigated only ifn-37 degrees exhibited antiviral act ...19892484831
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