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impact of gene vector design on the control of recombinant monoclonal antibody production by chinese hamster ovary cells.in this study, we systematically compare two vector design strategies for recombinant monoclonal antibody (mab) synthesis by chinese hamster ovary (cho) cells; a dual open reading frame (orf) expression vector utilizing separate cytomegalovirus (cmv) promoters to drive heavy chain (hc) and light chain (lc) expression independently, and a single orf vector design employing a single cmv promoter to drive hc and lc polypeptide expression joined by a foot and mouth disease virus f2a polypeptide self ...201121882365
Development of a serotype colloidal gold strip using monoclonal antibody for rapid detection type Asia1 foot-and-mouth disease.In this study, we developed a rapid, one step colloid gold strip (CGS) capable of specifically detecting type Asia1 foot-and-mouth disease virus (FMDV). We have produced two monoclonal antibodies (mAb) to type Asia1 FMD (named 1B8 and 5E2). On the test strip, the purified 1B8 labelled with the colloidal gold was used as the detector, and the purified 5E2 and goat anti-mouse antibodies were wrapped onto nitrocellulose (NC) membranes as the test and the control line, respectively. The rapid colloi ...201121880157
Role of macrophages in early protective immune responses induced by two vaccines against foot and mouth disease.Foot and Mouth Disease (FMD) is an acute disease of cloven-hoofed species. We studied the protection and early immune response induced in the murine model by vaccines formulated with inactivated virus and two different adjuvants. The presence of IMS12802PR or ISA206VG adjuvants yielded protection against viral challenge at early times post vaccination and induced FMDV-specific, but non neutralizing, antibody titers. In vivo macrophage depletion in vaccinated mice severely decreased the protectio ...201121878353
cpg oligodeoxynucleotide and montanide isa 206 adjuvant combination augments the immune responses of a recombinant fmdv vaccine in cattle.foot-and-mouth disease (fmd) is a highly contagious disease of cloven-hoofed animals. to prevent the spread of fmdv, inactivated virus vaccines are used to immunize animals in developing countries. however, there are safety concerns. in addition, it is difficult to distinguish the vaccinated animals from those naturally infected ones. in our lab, we have developed a recombinant fmdv vaccine named a7. a7 contained multiple b cell and t cell epitopes, which reside in a capsid protein (vp1) of fmdv ...201121872635
Modulation of Cytokine mRNA Expression in Pharyngeal Epithelial Samples obtained from Cattle Infected with Foot-and-Mouth Disease Virus.A novel technique of endoscopical collection of small tissue samples was used to obtain sequential tissue samples from the dorsal soft palate (DSP) of individual cattle infected with foot-and-mouth disease virus (FMDV) at different phases of the infection. Levels of mRNA encoding interferon (IFN)-a and IFN-ß as well as tumour necrosis factor (TNF)-a were measured in these samples by quantitative reverse transcriptase polymerase chain reaction. Expression of IFN-ß mRNA was significantly down-regu ...201121868026
recombinant non-structural polyprotein 3ab-based serodiagnostic strategy for fmd surveillance in bovines irrespective of vaccination.in india, the proportion of bovines vaccinated against foot-and-mouth disease (fmd) is increasing since the implementation of the government supported 'fmd control programme', and non-structural protein (nsp)-based serological assays for discriminating between antibodies induced by infection or vaccination (diva) could be useful. the fmd virus nsp 3ab was expressed in a prokaryotic system and an indirect elisa (r3ab(3) i-elisa) was developed and validated as a screening assay for detecting virus ...201121864578
develope monoclonal antibody against foot-and-mouth disease virus a type.in order to develop an anti-fmdv a type monoclonal antibody (mab), babl/c mice were immunized with fmdv a type. monoclonal antibodies (mabs) 7b11 and 8h4 against foot-and-mouth disease virus (fmdv) serotype a were produced by fusing sp2/0 myeloma cells with splenocyte from the mouse immunized with a/av88. the microneutralization titer of the mabs 7b11 and 8h4 were 1024 and 512, respectively. both mabs contain kappa light chains, the mabs were igg1. in order to define the mabs binding epitopes, t ...201121847759
reanalysis of the start of the uk 1967 to 1968 foot-and-mouth disease epidemic to calculate airborne transmission probabilities.the aims of this study were to statistically reassess the likelihood that windborne spread of foot-and-mouth disease (fmd) virus (fmdv) occurred at the start of the uk 1967 to 1968 fmd epidemic at oswestry, shropshire, and to derive dose-response probability of infection curves for farms exposed to airborne fmdv. to enable this, data on all farms present in 1967 in the parishes near oswestry were assembled. cases were infected premises whose date of appearance of first clinical signs was within ...201121846685
detection of foot-and-mouth disease virus rna in pharyngeal epithelium biopsy samples obtained from infected cattle: investigation of possible sites of virus replication and persistence.foot-and-mouth disease (fmd) is a highly contagious viral infection of significant financial importance to the export and trade of agricultural products. the occurrence of persistently infected "carriers" of fmd-virus (fmdv) in ruminant species adds further complications to disease control. there have been significant discrepancies in reports regarding the pathogenesis of fmdv infection in cattle with specific emphasis on the anatomical sites involved in early and persistent virus replication. i ...201121831538
recombinant vp1, an akt inhibitor, suppresses progression of hepatocellular carcinoma by inducing apoptosis and modulation of ccl2 production.the application of viral elements in tumor therapy is one facet of cancer research. recombinant capsid protein vp1 (rvp1) of foot-and-mouth disease virus has previously been demonstrated to induce apoptosis in cancer cell lines. here, we aim to further investigate its apoptotic mechanism and possible anti-metastatic effect in murine models of hepatocellular carcinoma (hcc), one of the most common human cancers worldwide.201121826248
Comparative study of codon substitution patterns in foot-and-mouth disease virus (serotype O).We compared genetic variations in the VP1 gene of foot-and-mouth disease viruses (FMDVs) isolated since 2000 according to the region of the world from. We analyzed relative synonymous codon usage (RSCU) and phylogenetic relationship between geographical regions, and calculated the genetic substitution patterns between Korean isolate and those from other countries. We calculated the ratios of synonymously substituted codons (SSC) to all observed substitutions and developed a new analytical parame ...201121825834
a dna vaccine encoding foot-and-mouth disease virus b and t-cell epitopes targeted to class ii swine leukocyte antigens protects pigs against viral challenge.development of efficient and safer vaccines against foot-and-mouth disease virus (fmdv) is a must. previous results obtained in our laboratory have demonstrated that dna vaccines encoding b and t cell epitopes from type c fmdv, efficiently controlled virus replication in mice, while they did not protect against fmdv challenge in pigs, one of the fmdv natural hosts. the main finding of this work is the ability to improve the protection afforded in swine using a new dna-vaccine prototype (pcmv-apc ...201121820470
evolutionary analysis of serotype a foot-and-mouth disease viruses circulating in pakistan and afghanistan during 2002-2009.foot-and-mouth disease (fmd) is endemic in pakistan and afghanistan. three different serotypes of the virus, namely o, a and asia-1, are responsible for the outbreaks of this disease in these countries. in the present study, the nucleotide (nt) coding sequences for the vp1 capsid protein (69 samples) or for all four capsid proteins (p1, 7 representative samples) of the serotype a fmd viruses circulating in pakistan and afghanistan were determined. phylogenetic analysis of the fmdv vp1 coding seq ...201121813704
mechanisms governing the selection of translation initiation sites on foot-and-mouth disease virus rna.translation initiation dependent on the foot-and-mouth disease virus internal ribosome entry site (ires) occurs at two sites (lab and lb), 84 nt apart. in vitro translation of an mrna comprising the ires and lab-lb intervening segment fused to a cat reporter has been used to study the parameters influencing the ratio of the two products and the combined product yield, as measures of relative initiation site usage and productive ribosome recruitment, respectively. with wild-type mrna ~40% of init ...201121813609
expression of foot and mouth disease virus nonstructural polyprotein 3abc with inactive 3c(pro) in escherichia coli.nonstructural 3abc protein of foot and mouth disease virus (fmdv) was widely used to differentiate vaccinated from natural fmdv-infected animals. 3abc is a polyprotein which is auto-processed to 3a, three copies of 3b and 3c(pro) by 3c(pro) protease. the 3abc gene was cloned and expressed in escherichia coli as native or mutated 3abc (mu3abc) forms. cysteine residues 142 and 163 of the catalytic triad within the 3c(pro) of mu3abc were changed to serine and glycine, respectively, to inhibit its p ...201121810474
combining livestock trade patterns with phylogenetics to help understand the spread of foot and mouth disease in sub-saharan africa, the middle east and southeast asia.international trade in animals and their products is recognised as a primary determinant of the global epidemiology of transboundary diseases such as foot and mouth disease (fmd). as well as causing serious production losses, fmd is highly contagious, being transmitted through multiple routes and hosts, which makes it one of the most important diseases affecting trade in livestock. its occurrence has dramatic consequences for the agricultural economy of a normally disease-free country, as well a ...201121809754
the codon usage model of the context flanking each cleavage site in the polyprotein of foot-and-mouth disease virus.to investigate the codon usage pattern of the contexts flanking 11 cleavage sites of foot-and-mouth disease virus (fmdv) polyprotein, the codon usage model of the corresponding codon position and the synonymous codon usage in the target contexts of 66 strains were characterized by two simple methods based on the relative synonymous codon usage value. the synonymous codons usage pattern was also compared between this virus and two species of hosts (cattle and domestic pig). it is indicated that f ...201121801856
proteomics analysis of porcine serum proteins by lc-ms/ms after foot-and- mouth disease virus (fmdv) infection.to analyze serum proteomics differences between normal and foot and mouth disease virus (fmdv)-infected piglets, an analytical method based on liquid chromatography with tandem mass spectrometry (lc-ms/ms) was used. samples of venous blood were collected before and after fmdv infection and high abundance serum albumin was removed using a commercial kit. after trypsin digestion, serum samples were processed with lc-ms/ms. proteins were identified by peptide mass fingerprinting. we found that apol ...201121799299
chemical disinfection of high-consequence transboundary animal disease viruses on nonporous surfaces.disinfection is a critical part of the response to transboundary animal disease virus (tadv) outbreaks by inactivating viruses on fomites to help control infection. to model the inactivation of tadv on fomites, we tested selected chemicals to inactivate foot and mouth disease virus (fmdv), african swine fever virus (asfv), and classical swine fever virus (csfv) dried on steel and plastic surfaces. for each of these viruses, we observed a 2 to 3 log reduction of infectivity due to drying alone. w ...201121798759
enterovirus 71 and coxsackievirus a16 3c proteases: binding to rupintrivir and their substrate, and anti-hfmd drug design.enterovirus 71 (ev71) and coxsackievirus a16 (cva16) are the major causative agents of hand foot and mouth disease (hfmd) which is prevalent in asia. thus far there are no prophylactic or therapeutic measures against hfmd. the 3c proteases from ev71 and cva16 play important roles in viral replication and are therefore ideal drug targets. by using biochemical, mutational and structural approaches, we broadly characterized both proteases. a series of high-resolution structures of the free or subst ...201121795339
the complete nucleotide sequences of serotype 'o' and 'a' indian vaccine strains of foot-and-mouth disease virus.the primary genomic structure of indian foot-and-mouth disease virus (fmdv) type a and o vaccine strains was determined, and sequences coding for individual proteins were compared with those of several complete genomic sequences. the clustering based on the whole genome was reflected by that observed with vp1 alone. the indian vaccine strains clustered with currently circulating regional strains, and type a and type o vaccine seeds showed close nucleotide similarity with a22iraq and o1manisa, re ...201121779906
translation of viral mrna without active eif2: the case of picornaviruses.previous work by several laboratories has established that translation of picornavirus rna requires active eif2a for translation in cell free systems or after transfection in culture cells. strikingly, we have found that encephalomyocarditis virus protein synthesis at late infection times is resistant to inhibitors that induce the phosphorylation of eif2a whereas translation of encephalomyocarditis virus early during infection is blocked upon inactivation of eif2a by phosphorylation induced by a ...201121779397
a brief review on diagnosis of foot-and-mouth disease of livestock: conventional to molecular tools.foot-and-mouth disease (fmd) is one of the highly contagious diseases of domestic animals. effective control of this disease needs sensitive, specific, and quick diagnostic tools at each tier of control strategy. in this paper we have outlined various diagnostic approaches from old to new generation in a nutshell. presently fmd diagnosis is being carried out using techniques such as virus isolation (vi), sandwich-elisa (s-elisa), liquid-phase blocking elisa (lpbe), multiplex-pcr (m-pcr), and ind ...201121776357
differential effects of viral vectors on migratory afferent lymph dendritic cells in vitro predicts enhanced immunogenicity in vivo.targeting dendritic cells (dc) is key to driving effective immune responses. lymphatic cannulation provides access to the heterogeneous populations of dc draining peripheral sites in rodents and ruminants. afferent lymph dec-205(+)cd11c(+)sirpa(+) dc were preferentially infected ex vivo with three vaccine viral vectors: replication defective human adenovirus-5 (rhuadv5); modified vaccinia virus ankara (rmva) and fowlpox virus (rfpv) all expressing green fluorescent protein (gfp). the rhuadv5 inf ...201121752909
structural basis for the biological relevance of the invariant apical stem in ires-mediated translation.rna structure plays a fundamental role in internal initiation of translation. picornavirus internal ribosome entry site (ires) are long, efficient cis-acting elements that recruit the ribosome to internal mrna sites. however, little is known about long-range constraints determining the ires rna structure. here, we sought to investigate the functional and structural relevance of the invariant apical stem of a picornavirus ires. mutation of this apical stem revealed better performance of g:c compa ...201121742761
understanding foot-and-mouth disease virus early pathogenesis and immune responses. 201121733132
detection of foot-and-mouth disease virus in the breath of infected cattle using a hand-held device to collect aerosols.exhaled air of individual cattle infected experimentally with foot-and-mouth disease virus (fmdv) was sampled to assess the feasibility of a rapid, non-invasive general screening approach for identifying sources of fmdv infection. the air sampler used was a handheld prototype device employing electrostatic particle capture in a microchip chamber of 10-15╬╝l and was shown to effectively capture a high percentage of airborne microorganisms. the particles were eluted subsequently from the chip cham ...201121723882
in-vitro and in-vivo phenotype of type asia 1 foot-and-mouth disease viruses utilizing two non-rgd receptor recognition sites.foot-and-mouth disease virus (fmdv) uses a highly conserved arg-gly-asp (rgd) triplet for attachment to host cells and this motif is believed to be essential for virus viability. previous sequence analyses of the 1d-encoding region of an fmdv field isolate (asia1/js/cha/05) and its two derivatives indicated that two viruses, which contained an arg-asp-asp (rdd) or an arg-ser-asp (rsd) triplet instead of the rgd integrin recognition motif, were generated serendipitously upon short-term evolution ...201121711567
[foot and mouth disease virus keeps lurking around the corner]. 201121702179
predicting antigenic sites on the foot-and-mouth disease virus capsid of the south african territories (sat) types using virus neutralization data.foot-and-mouth disease virus (fmdv) outer capsid proteins 1b, 1c and 1d contribute to the virus serotype distribution and antigenic variants that exist within each of the seven serotypes. this study presents phylogenetic, genetic and antigenic analyses of south african territories (sat) serotypes prevalent in sub-saharan africa. here, we show that the high levels of genetic diversity in the p1-coding region within the sat serotypes are reflected in the antigenic properties of these viruses and t ...201121697350
in silico analysis of ires rnas of foot-and-mouth disease virus and related picornaviruses.foot-and-mouth disease virus (fmdv) uses an internal ribosome entry site (ires), a highly structured segment of its genomic rna, to hijack the translational apparatus of an infected host. computational analysis of 162 type ii picornavirus ires rna sequences yielded secondary structures that included only base pairs supported by comparative or experimental evidence. the deduced helical sections provided the foundation for a hypothetical three-dimensional model of fmdv ires rna. the model was furt ...201121681504
immune potential of a novel multiple-epitope vaccine to fmdv type asia 1 in guinea pigs and sheep.to develop a safe and efficient recombinant subunit vaccine to foot-and-mouth disease virus (fmdv) type asia 1 in sheep, a tandem repeated multiple-epitope gene consisting of residues 137-160 and 197-211 of the vp1 gene of fmdv was designed and artificially synthesized. the biologically functional molecule, the ovine igg heavy constant region (oigg) as a protein carrier was introduced for design of the multiple-epitope recombinant vaccine and recombinant expression plasmids pet-30a-re and pet-30 ...201121667339
effective inhibition of foot-and-mouth disease virus (fmdv) replication in vitro by vector-delivered micrornas targeting the 3d gene.foot-and-mouth disease virus (fmdv) causes an economically important and highly contagious disease of cloven-hoofed animals. rnai triggered by small rna molecules, including sirnas and mirnas, offers a new approach for controlling viral infections. there is no report available for fmdv inhibition by vector-delivered mirna, although mirna is believed to have more potential than sirna. in this study, the inhibitory effects of vector-delivered mirnas targeting the 3d gene on fmdv replication were e ...201121663611
foot and mouth disease (fmd) virus: quantification of whole virus particles during the vaccine manufacturing process by size exclusion chromatography.foot and mouth disease (fmd) is a highly infectious viral disease that affects cattle, sheep, goats and swine causing severe economic losses worldwide. the efficacy of inactivated vaccines is critically dependent on the integrity of foot and mouth disease virus (fmdv) particles. the recommended method to quantify the active ingredient of vaccines is the 140s quantitative sucrose density gradient analysis. this method has been an immensely valuable tool over the past three decades but it is highl ...201121651939
[expression and utilization of 3ab nonstructural protein of foot-and-mouth disease virus in escherichia coli].to develop a sensitive and specific elisa for detection of antibodies to the nonstructural protein of fmdv. we cloned and expressed fmdv nonstructural protein 3ab in escherichia coli expression system. the recombinant protein 3ab was purified with ni-nta hisbind resins and characterized by western blotting. an indirect elisa based on purified protein 3ab as a coating antigen was established. the specificity and sensitivity of this assay were evaluated by comparison with a commercial 3abc-elisa k ...201121648221
reducing animal experimentation in foot-and-mouth disease vaccine potency tests.the world organisation for animal health (oie) terrestrial manual and the european pharmacopoeia (ep) still prescribe live challenge experiments for foot-and-mouth disease virus (fmdv) immunogenicity and vaccine potency tests. however, the ep allows for other validated tests for the latter, and specifically in vitro tests if a "satisfactory pass level" has been determined; serological replacements are also currently in use in south america. much research has therefore focused on validating both ...201121640777
enhancement of astragalus polysaccharide on the immune responses in pigs inoculated with foot-and-mouth disease virus vaccine.the effects of astragalus polysaccharides (aps) on the immune response in pigs immunized with foot-and-mouth disease virus (fmdv) vaccine were investigated. fifteen pigs were randomly divided into five groups. four groups were vaccinated with a fmdv inactivated vaccine. pigs in three experimental groups were administered varying doses of aps (aps1, 5mg/kg; aps2, 10mg/kg; aps3, 20mg/kg). the influence of aps on the number of cd3(+)cd4(-)cd8(+) cytotoxic t cells, cd3(+)cd4(+)cd8(+) t helper memory ...201121640133
validation of a recombinant integrin αvβ6/monoclonal antibody based antigen elisa for the diagnosis of foot-and-mouth disease.a sandwich elisa using recombinant integrin αvβ6 as a capture ligand and serotype-specific monoclonal antibodies (mabs) as detecting reagents has been compared with a polyclonal antibody based elisa (using type-specific rabbit antibodies as capture and guinea pig antibodies as detectors), which is employed routinely at the fao world reference laboratory for foot-and-mouth disease (fmd), for the identification and serotyping of fmd virus (fmdv). the study used cell culture grown antigens (1351 fm ...201121635921
virus inactivation by salt (nacl) and phosphate supplemented salt in a 3d collagen matrix model for natural sausage casings.due to possible presence and spread of contagious animal viruses via natural sausage casings the international trade in these food products is subject to veterinary and public health requirements. in order to manage these restrictions we determined the effect of casing preservation on four highly contagious viruses for livestock: foot-and-mouth-disease virus (fmdv), classical swine fever virus (csfv), swine vesicular disease virus (svdv) and african swine fever virus (asfv). we used an in vitro ...201121632134
outbreak of hand, foot and mouth disease caused by coxsackie a16 virus in a childcare centre in croatia, february to march 2011. 201121632018
genes controlling vaccine responses and disease resistance to respiratory viral pathogens in cattle.farm animals remain at risk of endemic, exotic and newly emerging viruses. vaccination is often promoted as the best possible solution, and yet for many pathogens, either there are no appropriate vaccines or those that are available are far from ideal. a complementary approach to disease control may be to identify genes and chromosomal regions that underlie genetic variation in disease resistance and response to vaccination. however, identification of the causal polymorphisms is not straightforw ...201121621277
multiplex rt-pcr detection and microarray typing of vesicular disease viruses.a vesicular disease multiplex reverse transcription (rt)-pcr with an accompanying microarray assay was developed for simultaneous detection and typing of foot-and-mouth disease virus (fmdv) and vesicular stomatitis virus (vsv), and for the detection of swine vesicular disease virus (svdv) and vesicular exanthema of swine virus (vesv). the multiplex rt-pcr successfully detected viral rna from a collection of 49 strains of vesicular viruses, including multiple strains from all seven serotypes of f ...201121620898
rapid screening swine foot-and-mouth disease virus using micro-elisa system.in order to tackle both regional and global foot-and-mouth disease virus (fmdv) epdimics, we hereby develop a rapid microfluidic thermal lens microscopic method to screen swine type o fmdv with good efficiency. the scheme has great merits in terms of field portability, sample volume, assay time, analytical sensitivity, and test reproducibility.201121617821
the importance of fmdv localisation in lymphoid tissue.foot-and-mouth disease virus, a highly contagious pathogen that can cause lameness, low weight and decreased milk production, is a scourge of agricultural livestock around the world. although the acute phase of infection is rarely fatal, infection may persist in animals that have apparently recovered, creating a viral reservoir that some fear could contribute to the spread of disease. we have used an array of molecular techniques to search for traces of virus in tissues from the mouths and throa ...201121616546
[diversity of receptor recognition site of type asial foot-and-mouth disease virus].foot-and-mouth disease virus (fmdv) initiates infection by binding to integrin receptors via a highly conserved arg-gly-asp (rgd) sequence found in the g-h loop of the structural protein vp1. however, fmdv is an rna virus, which harbors the evolutionary potential to render the rgd motif dispensable upon changes in constant environment. we studied mutation of rgd motif upon short-time passages fmdv asial/js/china/2005 field strain in different host.201121604558
comparative complete genome analysis of indian type a foot-and-mouth disease virus field isolates.comparative complete genome analysis of 17 serotype a indian field isolates representing different genotypes and sub-lineages is presented in this report. overall 79% of amino acids were invariant in the coding region. chunk deletion of nucleotide was observed in s and l fragment of 5'-utr. more variability which is comparable to that of capsid coding region was found in l and 3a region. functional motifs and residues critical for virus biology were conserved most. polyprotein cleavage sites acc ...201121604149
high cleavage efficiency of a 2a peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice.when expression of more than one gene is required in cells, bicistronic or multicistronic expression vectors have been used. among various strategies employed to construct bicistronic or multicistronic vectors, an internal ribosomal entry site (ires) has been widely used. due to the large size and difference in expression levels between genes before and after ires, however, a new strategy was required to replace ires. a self-cleaving 2a peptide could be a good candidate to replace ires because o ...201121602908
control of foot-and-mouth disease. 201121602410
phylogenetic analysis of foot-and-mouth disease virus type o circulating in the andean region of south america during 2002-2008.at present, foot-and-mouth disease (fmd) has been successfully controlled in most territories of south america, where only ecuador and venezuela remain as endemic countries. in this context, the precise characterization of circulating viruses is of utmost importance. this work describes the first molecular epidemiology study performed with the complete vp(1)-coding region of 114 field isolates of fmd virus (fmdv) type o, collected in the andean countries mainly during 2002-2008. sequences were a ...201121601999
analysis of the acute phase responses of serum amyloid a, haptoglobin and type 1 interferon in cattle experimentally infected with foot-and-mouth disease virus serotype o.abstract: a series of challenge experiments were performed in order to investigate the acute phase responses to foot-and-mouth disease virus (fmdv) infection in cattle and possible implications for the development of persistently infected "carriers". the host response to infection was investigated through measurements of the concentrations of the acute phase proteins (apps) serum amyloid a (saa) and haptoglobin (hp), as well as the bioactivity of type 1 interferon (ifn) in serum of infected anim ...201121592356
foot and mouth disease virus polyepitope protein produced in bacteria and plants induces protective immunity in guinea pigs.the goal of this project was to develop an alternative foot and mouth disease (fmd) vaccine candidate based on a recombinant protein consisting of efficient viral epitopes. a recombinant gene was designed that encodes b-cell epitopes of proteins vp1 and vp4 and t-cell epitopes of proteins 2c and 3d. the polyepitope protein (h-pe) was produced in e. coli bacteria or in n. benthamiana plants using a phytovirus expression system. the methods of extraction and purification of h-pe proteins from bact ...201121568869
[construction and expression of a coxsackievirus a16 vp1 gene plasmid which delivered by live attenuated salmonella.]aim: to develop a coxsackievirus a16 (cox a16) vp1 gene plasmid which delivered by live attenuated salmonella. methods: the plasmid which expressed vp1 protein of coxa16 was constructed by gene recombination. cellular expression was assessed by western bloten analysis. then the recombinant attenuated salmonella which harboring the plasmid were constructed by electro transformation. results: coxa16 vp1 gene sequence was inserted into a eukaryotic expression plasmid. vp1 protein was detected in th ...201121557905
relationship between clinical signs and transmission of an infectious disease and the implications for control.control of many infectious diseases relies on the detection of clinical cases and the isolation, removal, or treatment of cases and their contacts. the success of such "reactive" strategies is influenced by the fraction of transmission occurring before signs appear. we performed experimental studies of foot-and-mouth disease transmission in cattle and estimated this fraction at less than half the value expected from detecting virus in body fluids, the standard proxy measure of infectiousness. th ...201121551063
development of one-step multiplex rt-pcr method for simultaneous detection and differentiation of foot-and-mouth disease virus serotypes o, a, and asia 1 circulating in vietnam.a one-step multiplex rt-pcr method using new primers was developed for the simultaneous detection and differentiation of vietnamese fmdv serotypes o, a, and asia 1 directly from clinical samples. the rt-pcr method used a cocktail of one universal minus-sense primer designed in the 2b gene and three serotype-specific plus-sense primers designed in the hypervariable regions of the capsid vp1 coding gene of fmdv. these serotype-specific primer pairs amplified 658, 535, and 427bp pcr products corres ...201121550367
identification of the role of rig-i, mda-5 and tlr3 in sensing rna viruses in porcine epithelial cells using lentivirus-driven rna interference.pathogen recognition receptors are essential for antiviral host immune responses. these specialized receptors detect conserved viral compounds and induce type i interferons (ifn) and pro-inflammatory cytokines. here we evaluated the contribution of rig-i, mda-5 and tlr3 to the recognition of classical swine fever (csfv), foot-and-mouth disease virus (fmdv), vesicular stomatitis virus (vsv) and influenza a virus (iav) to ifn-β responses in the porcine epithelial cell line pk-15. to this end, we i ...201121539869
an experimental infection in pigs using a foot-and-mouth disease virus isolated from the 2010 epidemic in japan.in this study, we carried out an experimental infection in pigs using a foot-and-mouth disease virus isolated from the 2010 epidemic in japan to analyze clinical manifestation, antibody response and virus sedding patterns in pigs. we found that the virus was virulent in pigs, producing a synchronous disease in the inoculated pigs and efficient spread to direct contact pigs. these results are useful for epidemiologically investigating the 2010 epidemic in japan and improving the measures for cont ...201121532260
rna structural domains in non-coding regions of foot-and-mouth disease virus genome trigger innate immunity in porcine cells and mice.induction of type i interferons (ifn-α/β) in response to viral infection is a crucial step leading to the antiviral state in the host. viruses produce dsrna during their replication cycle that is sensed as non-self by host cells through different receptors. a signalling cascade is then activated to block viral replication and spread. foot-and-mouth disease virus (fmdv) is a picornavirus, highly sensitive to ifn, causing one of the world's most important animal diseases. in this study, we showed ...201121525336
development and validation of a prokaryotically expressed foot-and-mouth disease virus non-structural protein 2c'3ab-based immunochromatographic strip to differentiate between infected and vaccinated animals.abstract:201121513550
evidence of activation and suppression during the early immune response to foot-and-mouth disease virus.foot-and-mouth disease virus causes a serious disease of livestock species, threatening free global trade and food security. the disease spreads rapidly between animals, and to ensure a window of opportunity for such spread, the virus has evolved multiple mechanisms to subvert the early immune response. the cycle of infection in the individual animal is very short, infection is initiated, disseminated throughout the body and infectious virus produced in <7 days. foot-and-mouth disease virus has ...201121501424
isolation, identification and complete genome sequence analysis of a strain of foot-and-mouth disease virus serotype asia1 from pigs in southwest of china.abstract: backgroud: foot-and-mouth disease virus (fmdv) serotype asia1 generally infects cattle and sheep, while its infection of pigs is rarely reported. in 2005-2007, fmd outbreaks caused by asia1 type occurred in many regions of china, as well as some parts of east asia countries. during the outbreaks, there was not any report that pigs were found to be clinically infected. results: in this study, a strain of fmdv that isolated from pigs was identified as serotype asia1, and designated as "a ...201121496298
biosecurity and fmd transmission. 201121493485
establishment of persistent infection with foot-and-mouth disease virus in bhk-21 cells.abstract:201121492421
[current situation and future preventive measures of foot-and-mouth disease in japan].foot-and-mouth disease caused by foot-and-mouth disease virus (fmdv) is a severe and acute vesicular disease of cloven-hoofed animals including cattle, pigs, sheep and goats. as fmdv is highly contagious and causes productivity losses among infected animals, outbreaks of the disease are a primary animal health concern worldwide. in april, 2010, the disease reoccurred in miyazaki prefecture in 10 years. compared to the outbreak in 2000 in which no infection among pigs was observed, a total of 292 ...201021488337
comparative sequence analysis of representative foot-and-mouth disease virus genomes from southeast asia.foot-and-mouth disease (fmd) is endemic in mainland southeast asia where up to seven genetically distinct viral lineages co-circulate (o/sea/mya-98, o/sea/cam-94, o/me-sa/panasia, o/me-sa/panasia-2, o/cathay, a/asia/sea-97, and serotype asia 1). the aim of this study was to analyse the sequence variability between representative complete genomes for these seven lineages. the genome sequences varied from 8130 to 8192 nucleotides in length and shared nucleotide identities ranging from 91.8 to 78.9 ...201121479678
closed ampoule isothermal microcalorimetry for continuous real-time detection and evaluation of cultured mammalian cell activity and responses.closed ampoule isothermal microcalorimetry (imc) is a simple, powerful, nondestructive, and convenient technique that allows continuous, real-time detection and evaluation of cultured cell activity and responses. at a selected set temperature, imc measures the heat flow between a sample and a heat sink and compares it to the heat-flow between a thermally inactive reference and the heat sink. since heat flow rates are proportional to the rates of chemical reactions and changes of state, imc provi ...201121468980
differentiation of foot-and-mouth disease-infected pigs from vaccinated pigs using antibody-detecting sandwich elisa.the presence of serum antibodies for nonstructural proteins of the foot-and-mouth disease virus (fmdv) can differentiate fmdv-infected animals from vaccinated animals. in this study, a sandwich elisa was developed for rapid detection of the foot-and-mouth disease (fmd) antibodies; it was based on an escherichia coli-expressed, highly conserved region of the 3abc nonstructural protein of the fmdv o/tw/99 strain and a monoclonal antibody derived from the expressed protein. the diagnostic sensitivi ...201121467761
viewpoint: factors involved in type i interferon responses during porcine virus infections.since type i interferon (ifn-i) is considered a potent antiviral defence mechanism, it is not surprising that during evolution viruses have development of various mechanisms to counteract ifn-i induction or release. despite this, certain virus infections are associated with very high levels of systemic ifn-i. one explanation for this observation is the presence of plasmacytoid dendritic cells (pdc), which are able to produce high levels of ifn-i despite the presence of viral ifn-i antagonists. e ...201121458079
a highly sensitive detection for foot-and-mouth disease virus by gold nanopariticle improved immuno-pcr.abstract: background: foot-and-mouth disease (fmd) is one of the most contagious of all artiodactyl animal diseases, and its infection has an obvious ability to spread over long distances and to contribute to epidemics in fmd-free areas. a highly sensitive and specfic method is required to detect fmdv. in this study, we evaluated the usefulness of a bio-barcode assay (bca) technique for detecting clinical samples of fmdv. methods: highly sensitive gold nanopariticle (gnp) improved immuno -pcr (g ...201121453461
skin as a potential source of infectious foot and mouth disease aerosols.this review examines whether exfoliated, virus-infected animal skin cells could be an important source of infectious foot and mouth disease virus (fmdv) aerosols. infectious material rafting on skin cell aerosols is an established means of transmitting other diseases. the evidence for a similar mechanism for fmdv is: (i) fmdv is trophic for animal skin and fmdv epidermis titres are high, even in macroscopically normal skin; (ii) estimates for fmdv skin cell aerosol emissions appear consistent wi ...201121450741
phylogenomics and molecular evolution of foot-and-mouth disease virus.this report describes the use of bayesian methods to analyze polyprotein coding region sequences (n = 217) obtained from genbank to define the genome-wide phylogeny of foot and mouth disease virus (fmdv). the results strongly supported the monophyly of five fmdv serotypes, o, a, asia 1, c, and sat 3, while sequences for the two remaining fmdv serotypes, sat 1 and sat 2 did not separate into entirely distinct clades. the phylogenomic tree revealed three sister-group relationships, serotype o + as ...201121448588
viral genome segmentation can result from a trade-off between genetic content and particle stability.the evolutionary benefit of viral genome segmentation is a classical, yet unsolved question in evolutionary biology and rna genetics. theoretical studies anticipated that replication of shorter rna segments could provide a replicative advantage over standard size genomes. however, this question has remained elusive to experimentalists because of the lack of a proper viral model system. here we present a study with a stable segmented bipartite rna virus and its ancestor non-segmented counterpart, ...201121437265
antiviral activity of bovine type iii interferon against foot-and-mouth disease virus.foot-and-mouth disease (fmd) is one of the most serious threats to the livestock industry. despite the availability of a vaccine, recent outbreaks in disease-free countries have demonstrated that development of novel fmd control strategies is imperative. here we report the identification and characterization of bovine (bo) interferon lambda 3 (ifn-++3), a member of the type iii ifn family. expression of boifn-++3 using a replication-defective human adenovirus type 5 vector (ad5-boifn-++3) yielde ...201121435672
development of vaccines toward the global control and eradication of foot-and-mouth disease.foot-and-mouth disease (fmd) is one of the most economically and socially devastating diseases affecting animal agriculture throughout the world. although mortality is usually low in adult animals, millions of animals have been killed in efforts to rapidly control and eradicate fmd. the causing virus, fmd virus (fmdv), is a highly variable rna virus occurring in seven serotypes (a, o, c, asia 1, sat 1, sat 2 and sat 3) and a large number of subtypes. fmdv is one of the most infectious agents kno ...201121434805
florfenicol impairs the immune responses to vaccination against foot-and-mouth disease in mice.florfenicol is a new type of broad-spectrum antibacterial that has been used in veterinary clinics. it showed immunosuppressive activity on the immune responses to vaccination against foot-and-mouth disease virus (fmdv) serotype o in mice. in the present study, balb/c mice were immunized subcutaneously with fmdv serotype o antigen on days 1 and 14. beginning on day 21, mice were treated with a single daily oral dose of florfenicol (50, 100, and 200?mg/kg) for seven consecutive days. on day 28, b ...201121428714
5-fluorouracil in lethal mutagenesis of foot-and-mouth disease virus.5-fluorouracil (fu) is a pyrimidine analogue extensively used in cancer chemotherapy. fu can be metabolized into 5-fluorouridine-triphosphate, which can be used as substrate for viral rna-dependent rna polymerases. this results in the incorporation of mutations into viral rna. accumulation of mutations may lead to loss of virus infectivity, in a process known as lethal mutagenesis. rna virus pathogens are particularly difficult to control because they are highly mutable, and mutants resistant to ...200921426129
genetic diversity of foot-and-mouth disease virus serotype o in pakistan and afghanistan, 1997-2009.foot-and-mouth disease (fmd) is endemic in pakistan and afghanistan; serotypes o, a and asia-1 of the virus are responsible for the outbreaks in these countries with fmdv type o usually being the most common. in the present study, the nucleotide sequences encoding the fmdv capsid protein vp1 from virus samples were determined. phylogenetic analysis of the serotype o fmd viruses circulating in pakistan and afghanistan between 1997 and 2009 revealed the presence of at least three different lineage ...201121419880
pan-serotypic detection of foot-and-mouth disease virus using a minor groove binder probe reverse transcription polymerase chain reaction assay.a novel assay for the pan-serotypic detection of foot-and-mouth disease virus (fmdv) was designed using a 5' conjugated minor groove binder (mgb) probe real-time rt-pcr system. this assay targets the 3d region of the fmdv genome and is capable of detecting 20 copies of a transcribed rna standard. the linear range of the test was eight logs from 2 × 10¹ to 2 × 108 copies and amplification time was approximately 2 h. using a panel of 83 rna samples from representative fmdv isolates, the diagnostic ...201121419170
foot-and-mouth disease in feral swine: susceptibility and transmission.experimental studies of foot-and-mouth disease (fmd) in feral swine are limited, and data for clinical manifestations and disease transmissibility are lacking. in this report, feral and domestic swine were experimentally infected with fmdv (a24-cruzeiro), and susceptibility and virus transmission were studied. feral swine were proved to be highly susceptible to a-24 cruzeiro fmd virus by intradermal inoculation and by contact with infected domestic and feral swine. typical clinical signs in fera ...201121418546
2a to the fore - research, technology and applications.the 2a region of the foot-and-mouth disease virus (fmdv) encodes a short sequence that mediates self-processing by a novel translational effect. translation elongation arrest leads to release of the nascent polypeptide and re-initiation at the next in-frame codon. in this way discrete translation products are derived from a single open reading frame. active 2a-like sequences have been found in (many) other viruses and trypanosome non-ltr retrotransposons. exponential growth of 2a technology with ...201021415883
foot-and-mouth disease virus leader proteinase inhibits dsrna-induced rantes transcription in pk-15 cells.the chemokine rantes (regulated upon activation, normal t-cells expressed and secreted) plays an essential role in inflammation and immune response. infection with wild-type foot-and-mouth disease virus (fmdv) in pk-15 cells strongly inhibits the expression of rantes compared to infection with a genetically engineered mutant lacking the leader protein (l(pro)) coding region. this suggests that l(pro) is involved in rantes regulation. however, the underlying molecular mechanism remains unclear. i ...201121399922
the importance of being short: the role of rabies virus phosphoprotein isoforms assessed by differential ires translation initiation.the rabies virus (rv) phosphoprotein p is a multifunctional protein involved in viral rna synthesis and in counteracting host innate immune responses. we have previously shown that rv p gene expression levels can be regulated by using picornavirus internal ribosome entry site (ires) elements. here we exploited a particular feature of the foot-and-mouth disease virus (fmdv) ires, namely, preferential initiation at a downstream initiation codon, to address the role of n-terminally truncated rv pho ...201121397980
development of expression vectors based on pepino mosaic virus. 201121396092
absence of heat intolerance (panting) syndrome in foot-and-mouth disease-affected indian cattle (bos indicus) is associated with intact thyroid gland function.foot-and-mouth disease (fmd) is a highly contagious and economically important viral disease with high morbidity and reduced productivity of affected animals. we studied the heat intolerance (hi) (panting) syndrome and the effect of fmd virus (fmdv) infection on thyroid gland function in indian cattle (bos indicus). experimental infection with fmdv asia 1 resulted in a mild form of disease with superficial lesions. heat intolerance syndrome and its signs were not observed among the recovered ani ...201121388520
rapid and transient activation of +¦+¦ t cells to ifn-+¦ production, nk cell-like killing, and antigen processing during acute virus infection.+¦+¦ t cells are the majority peripheral blood t cells in young cattle. the role of +¦+¦ t cells in innate responses against infection with foot-and-mouth disease virus was analyzed on consecutive 5 d following infection. before infection, bovine wc1(+) +¦+¦ t cells expressed a nonactivated phenotype relative to cd62l, cd45ro, and cd25 expression and did not produce ifn-+¦ ex vivo. additionally, cd335 expression was lacking and no spontaneous target cell lysis could be detected in vitro, althoug ...201121383249
fine mapping of a foot-and-mouth disease virus epitope recognized by serotype-independent monoclonal antibody 4b2.vp2 is a structural protein of the foot-and-mouth disease virus (fmdv). in this study, a fmdv serotype-in-dependent monoclonal antibody (mab), 4b2, was generated. by screening a phage-displayed random 12-peptide library, we found positive phages displaying the consensus motif ettxle (x is any amino acid (aa)), which is highly homologous to (6)ettlle(11) at the n-terminus of the vp2 protein. subsequently, a series of gst-fusion proteins expressing a truncated n-terminus of vp2 were examined by we ...201121369985
the expressions of hsp70 and ab-crystallin in myocarditis associated with foot-and-mouth disease virus in lambs.this study describes the expression of heat shock protein70 (hsp70) and alpha-basic-crystallin (a-bc) and their association with apoptosis and some related adaptor proteins in the pathogenesis of foot-and-mouth disease virus (fmdv)-induced myocarditis in lambs. hsp70 was generally overexpressed in the myocardial tissues and inflammatory cells of fmdv-induced myocarditis with differential accumulation and localization in same hearts when compared to non-foot-and-mouth disease control hearts. a-bc ...201121368565
synthetic peptides.efforts to produce more stable and defined vaccines have concentrated on studying, in detail, the immune response to many infectious diseases in order to identify the antigenic sites on the pathogens that are involved in stimulating protective immumty. armed with this knowledge, it is possible to mimic such sites by producing short chains of amino acids (peptides) and to use these as the basis for novel vaccines. the earliest documented work on peptide immunization is actually for a plant virus, ...199621359696
a mutant of asia 1 serotype of foot-and-mouth disease virus with the deletion of an important antigenic epitope in the 3a protein.foot-and-mouth disease is a highly contagious viral disease of cloven-hoofed animals. the availability of a vaccine for differentiating infected from vaccinated animals (diva) is crucial for the control and eradication of foot-and-mouth disease virus (fmdv). because traditional inactivated vaccines may contain trace nonstructural proteins interfering with the diva, we hypothesized that mutant fmdv with deletion of immunodominant epitopes may be valuable. our previous study has generated a full-l ...201121358757
estimating risk factors for farm-level transmission of disease: foot and mouth disease during the 2001 epidemic in great britain.controlling an epidemic would be aided by establishing whether particular individuals in infected populations are more likely to transmit infection. however, few analyses have characterised such individuals. such analyses require both data on who infected whom and on the likely determinants of transmission; data that are available at the farm level for the 2001 foot and mouth disease epidemic in great britain. using these data a putative number of daughter infected premises (ips) resulting from ...201021352781
transmission risks and control of foot-and-mouth disease in the netherlands: spatial patterns.in 2001 the epidemics of foot-and-mouth disease virus (fmdv) in great britain, the netherlands and france have shown how fast fmdv may spread between farms. the massive socio-economic impact of these epidemics and the intervention measures taken demonstrate the need for quantitative assessments of the efficacy of candidate intervention strategies. here we use a mathematical model to describe the spatial transmission of fmdv in the netherlands and use the dutch 2001 outbreak data to estimate mode ...201021352775
development and initial results of a low cost, disposable, point-of-care testing device for pathogen detection.development of small footprint, disposable, fast, and inexpensive devices for pathogen detection in the field and clinic would benefit human and veterinary medicine by allowing evidence-based responses to future out breaks. we designed and tested an integrated nucleic acid extraction and amplification device employing a loop-mediated isothermal amplification (lamp) or reverse transcriptase-lamp assay. our system provides a screening tool with polymerase-chain-reaction-level sensitivity and speci ...201121342806
a pseudotype baculovirus expressing the capsid protein of foot-and-mouth disease virus and a t-cell immunogen shows enhanced immunogenicity in mice.foot-and-mouth disease (fmd) is a highly contagious disease of livestock which causes severe economic loss in cloven-hoofed animals. vaccination is still a major strategy in developing countries to control fmd. currently, inactivated vaccine of fmdv has been used in many countries with limited success and safety concerns. development of a novel effective vaccine is must.201121342530
experimental evidence for competitive growth advantage of genotype vii over vi: implications for foot-and-mouth disease virus serotype a genotype turnover in nature.in india, systematic genotype replacement has been observed for serotype a foot-and-mouth disease virus. after a decade of co-circulation of genotypes vi and vii, genotype vii emerged as the single dominant genotype since 2001. to derive possible explanations for such epochal evolution dynamics, in vitro intergenotype growth competition experiments involving both co- and superinfection regimes were conducted. coinfection of bhk-21 cells demonstrated abrupt loss in the genotype vi viral load with ...201121338995
a simple and rapid colloidal gold-based immunochromatogarpic strip test for detection of fmdv serotype a.a sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (fmdv) serotypes was developed. in this rapid test, affinity purified polyclonal antibodies from guinea pigs which were immunized with sucking-mouse adapted fmd virus (a/av88(l) strain) were conjugated to colloidal gold beads and used as the capture antibody, and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted fmd virus (a/cha/09 strain) were used as detect ...201121331888
genomic comparison of foot-and-mouth disease virus r strain and its chick-passaged attenuated strain.the present study examined the genomic differences between foot-and-mouth disease virus (fmdv) r strain and its attenuated, chick-passaged (r(304)) strain. eleven pairs of primers were used to amplify the complete genome of fmdv r and r(304) by rt-pcr. each fragment was cloned into pmd18-t vector and sequenced. nucleotide analyses showed that the genome encoding regions of r and r(304) strains open reading frame (orf) were both 6966 nucleotides (nt) in length, encoding 2322 amino acids. one hund ...201121330068
integrin sub-unit expression in cell cultures used for the diagnosis of foot-and-mouth disease.the ability to propagate foot-and-mouth disease virus (fmdv) plays an important role in laboratory diagnosis and the production of vaccines to control the spread of the disease. many established cell lines suffer from poor sensitivity for isolating virus from field samples. one possible factor that limits sensitivity to fmdv is the lack of expression of surface integrins, the primary class of cell receptor used by fmdv to initiate infection. in this study we have sequenced cdnas encoding these m ...201121329991
active protein aggregates induced by terminally attached self-assembling peptide elk16 in escherichia coli.in recent years, it has been gradually realized that bacterial inclusion bodies (ibs) could be biologically active. in particular, several proteins including green fluorescent protein, ß-galactosidase, ß-lactamase, alkaline phosphatase, d-amino acid oxidase, polyphosphate kinase 3, maltodextrin phosphorylase, and sialic acid aldolase have been successfully produced as active ibs when fused to an appropriate partner such as the foot-and-mouth disease virus capsid protein vp1, or the human ß-amylo ...201121320350
molecular characterization of foot-and-mouth disease virus: implications for disease control in bangladesh.foot-and-mouth disease (fmd) is endemic in bangladesh, and to implement an effective fmd control programme, it is essential to understand the complex epidemiology of the disease. here, we report on the characterization of fmd virus (fmdv) recovered from fmd outbreaks in bangladesh in late 2009. all isolated viruses belonged to the fmdv serotype o. the phylogenetic reconstruction showed that all isolates belonged to the middle east-south asia (me-sa) topotype, but fell into two distinct sublineag ...201121320294
effect of thiomersal on dissociation of intact (146s) foot-and-mouth disease virions into 12s particles as assessed by novel elisas specific for either 146s or 12s particles.intact (146s) foot-and-mouth disease virions (fmdvs) can dissociate into specific (12s) viral capsid degradation products. using two single-domain antibody fragments that bind specifically to either 146s or 12s particles we developed two elisas for the quantification of these particles in fmdv antigen preparations used for vaccine manufacturing. only o serotype strains are detected in the 146s specific elisa whereas strains of most serotypes are detected in the 12s specific elisa. however, the 1 ...201121316500
dna immunization of pigs with foot-and-mouth disease virus minigenes: from partial protection to disease exacerbation.despite several attempts to design new vaccines, there are as of yet no available alternatives to the conventional fmdv vaccines. here, we present the divergent results obtained in pigs after immunization with two experimental dna vaccines encoding one b and two t cell fmdv epitopes, either expressed alone (pcmv-btt) or fused to a strong signal peptide (pcmv-spbtt). while all pigs vaccinated with pcmv-spbtt showed both a delay in the disease onset and reduced severity of signs and lesions after ...201121315115
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