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PubMed2006

2'-C-methylcytidine as a potent and selective inhibitor of the replication of foot-and-mouth disease virus.

We report on the potent and selective in vitro antiviral activity of 2'-C-methylcytidine (2'-C-MetCyt) against foot-and-mouth disease virus (FMDV). FMDV belongs to the Picornaviridae and has the potential to cause devastating epidemics in livestock. The 50% and 90% effective concentrations (EC50 and EC90) for inhibition of the FMDV-induced cytopathic effect (CPE) formation were 6.4+/-3.8 and 10.8+/-5.4 microM. Comparable EC50 values for inhibition of viral RNA synthesis were observed. Treatment of FMDV-infected BHK-21 cells with 77 microM 2'-C-MetCyt resulted in a (1.6-3.2)x10(3)-fold reduction of infectious virus yield. Time-of-drug addition experiments suggest that 2'-C-MetCyt interacts with viral replication at a time point that coincides with the onset of intracellular viral RNA synthesis. In contrast to emergency vaccination, a potent and selective antiviral agent may provide almost immediate (prophylactic/therapeutic) protection against infection and thus constitute an important alternative/supplementary option to contain outbreaks such as those caused by FMDV.

Animals↗
PubMed2006

Foot-and-mouth disease virus 3C protease: recent structural and functional insights into an antiviral target.

The 3C protease from foot-and-mouth disease virus (FMDV 3C(pro)) is critical for viral pathogenesis, having vital roles in both the processing of the polyprotein precursor and RNA replication. Although recent structural and functional studies have revealed new insights into the mechanism and function of the enzyme, key questions remain that must be addressed before the potential of FMDV 3C(pro) as an antiviral drug target can be realised.

3C Viral Proteases↗
PubMed2005

Development of novel strategies to control foot-and-mouth disease: marker vaccines and antivirals.

Foot-and-mouth disease (FMD) is economically the most important viral-induced livestock disease worldwide. The disease is highly contagious and FMD virus (FMDV) replicates and spreads extremely rapidly. Outbreaks in previously FMD-free countries, including Taiwan, the United Kingdom, and Uruguay, and the potential use of FMDV by terrorist groups have demonstrated the vulnerability of countries and the need to develop control strategies that can rapidly inhibit or limit disease spread. The current vaccine, an inactivated whole virus preparation, has a number of limitations for use in outbreaks in disease-free countries. We have developed an alternative approach using a genetically engineered FMD subunit vaccine that only contains the portions of the viral genome required for virus capsid assembly and lacks the coding region for most of the viral nonstructural (NS) proteins including the highly immunogenic 3D protein. Thus, animals inoculated with this marker vaccine can readily be differentiated from infected animals using diagnostic assays employing the NS proteins not present in the vaccine and production of this vaccine, which does not contain infectious FMDV, does not require expensive high-containment manufacturing facilities. One inoculation of this subunit vaccine delivered in a replication-defective human adenovirus vector can induce rapid, within 7 days, and relatively long-lasting protection in swine. Similarly cattle inoculated with one dose of this recombinant vector are rapidly protected from direct and contact exposure to virulent virus. Furthermore, cattle given two doses of this vaccine developed high levels of FMDV-specific neutralizing antibodies, but did not develop antibodies against viral NS proteins demonstrating the ability of FMD subunit vaccinated animals to be differentiated from infected animals. To stimulate early protection prior to the vaccine-induced adaptive immune response we inoculated swine with the antiviral agent, type I interferon, and induced complete protection within 1 day. Protection can last for 3-5 days. The combination of the FMD marker vaccine and type I interferon can induce immediate, within 1 day, and long-lasting protection against FMD. Thus, this combination approach successfully addresses a number of concerns of FMD-free countries with the current disease control plan. By rapidly limiting virus replication and spread this strategy may reduce the number of animals that need to be slaughtered during an outbreak.

Adenoviruses, Human↗
PubMed2006

A metastatic haemangiopericytoma of the floor of the mouth.

A case of metastatic haemangiopericytoma in the floor of the mouth is described. Haemangiopericytoma is a relatively rare slow-growing vascular tumour with variable malignant potential. This tumour has been identified in almost every region of the body, but its occurrence in the oral cavity has been rarely reported. The rate of regional and distant metastasis of the tumour is low. This case, presented 12 years after initial surgery suggested the need for careful long-term follow-ups of patients with haemangiopericytoma.

Antigens, CD34↗
PubMed2004

Foregut duplication cyst arising in the floor of mouth.

Enteric duplications are uncommon. A rare foregut duplication cyst arising in the floor of mouth is reported. Diagnosis of cystic lesion within the oral cavity was made in utero. Enteric duplications have not yet been attributed to a single embryogenesis but are likely to represent an error in migration of normal cells that rest in an abnormal position. Immediately following delivery of the neonate via an ex-utero intrapartum treatment (EXIT) procedure, the cyst was marsupialised but required definitive surgical excision at 5 weeks of age.

Cysts↗
PubMed2004

Determinants of early foot-and-mouth disease virus dynamics in pigs.

This paper provides a quantitative description of the early infectious process in pigs experimentally infected with foot-and-mouth disease virus (FMDV), obtained by dose-dependent, time course studies of viral load in serum. Pigs were inoculated by the intravenous or intradermal/subcutaneous route with FMDV and housed together in groups or individually. The effects of dose, inoculation route and exposure intensity on the replication of FMDV in vivo and the development of disease were studied. It was shown that the higher the dose, the shorter was the time to the start of active viraemia and to the onset of clinical signs. Exposure intensity and housing conditions influenced the viral dynamics of FMDV. Increasing the exposure intensity, by increasing the number of infected pigs housed together, had the effect of synchronizing the infection and reducing the variance in the start of active viraemia. Increasing the number of pigs housed together also increased the interaction between the pigs and the activity of individual pigs, which had the effect of shortening the time to the onset of clinical signs such as vesicle formation. Intradermal inoculation was more effective than intravenous inoculation for transmitting FMDV to pigs, resulting in shorter times to the start of active viraemia and in higher clinical scores.

Animals↗
PubMed2004

Isolation of foot-and-mouth disease virus specific bovine antibody fragments from phage display libraries.

Foot-and-mouth disease virus (FMDV) is an important veterinary pathogen which can cause widespread epidemics. Due to the high antigenic variability of FMDV, it is important to undertake mutation analysis under immunological pressure. To study the bovine antibody response at a molecular level, phage display technology was used to produce bovine anti-FMDV Fabs. CH1-VH chains with FMDV specific binding could be isolated after selection from a library made from vaccinated cattle. Though their involvement in the bovine immune response remains to be ascertained, it is planned to express the five different selected VH domains in bacterial or insect systems as sequence homologies with integrin beta6 chain could shed light on the basis of FMDV type receptor specificities.

Amino Acid Sequence↗
PubMed2004

Validation of a foot-and-mouth disease antibody screening solid-phase competition ELISA (SPCE).

This paper describes the validation of a solid-phase competition enzyme-linked immunosorbent assay (SPCE) for the serological detection of antibody to serotype O foot-and-mouth disease (FMD) in sheep, cattle and pigs. The specificity of the SPCE was calculated from the results of testing known negative sera from sheep, cattle and pigs (n=3030, 1418 and 1495, respectively). The mean percentage inhibition (PI) for known negative sheep, cattle and pig sera were 19.3, 24.1 and 20.8%, respectively. The specificity of the SPCE at a cut-off point (COP) of 60 PI was 99.50% for sheep sera, 99.44% for cattle sera and 100% for pig sera. The analytical sensitivity of the SPCE was examined by testing sera from sheep, cattle and pigs. Based on the testing of serial bleeds from experimentally infected animals, seroconversion at the 60 PI COP occurred between 4 and 9 days post-infection or -exposure, similar to that observed using the virus neutralisation test (VNT) with a COP of 1/45. When applied to 267 sheep and 143 pig samples, that were obtained in Great Britain (GB) during the 2001 FMD UK outbreak, the SPCE identified more positive samples than did the VNT. Estimates of the accuracy, repeatability and reproducibility of the SPCE were verified during the large-scale serosurveillance necessitated by the 2001 outbreak. Results from field and experimental sera showed that when compared against the VNT, the sensitivity of the SPCE was less affected by the choice of virus strain used in the test. Using the O(1) UKG 2001 FMD virus in the VNT with samples representative of the uninfected GB sheep population, the test specificity was 100% at a COP of 1/45.

Animals↗
PubMed2004

Comparison of ELISA for the detection of porcine serum antibodies to non-structural proteins of foot-and-mouth disease virus.

Three foot-and-mouth disease virus non-structural protein antibody detection kits, CHEKIT FMD-3ABC, UBI FMD NS EIA and DVIVR NSP ELISA, were compared in the study. The results showed that the specificity of the kits ranged from 96.7 to 100% in nai;ve pigs and from 93.6 to 98.1% in vaccinated pigs, and that the DVIVR kit had the highest analytical sensitivity. The kappa statistics for the detection of 612 sera were 0.582, 0.447 and 0.658 for CHEKIT/UBI, CHEKIT/DVIVR and UBI/DVIVR, respectively. This study also revealed that measurable non-structural protein specific antibodies in some of infected pigs were sustained either for shorter periods or in intermittent patterns, thus aggravating the difficulties associated with the removal of pre-exposed pigs in the field.

Animals↗
PubMed2004

Development and use of a biotinylated 3ABC recombinant protein in a solid-phase competitive ELISA for the detection of antibodies against foot-and-mouth disease virus.

A biotinylated 3ABC recombinant protein was developed and used in a competitive ELISA (cELISA) to detect foot-and-mouth disease virus (FMDV) antibodies in cattle, sheep and pigs. In this report, we describe the cloning and expression of 3ABC protein in Escherichia coli cells as fusion protein with 6xHis and biotin. This cELISA uses streptavidin to capture bacterially expressed and in vivo biotinylated 3ABC antigen. The antigen capture strategy provides a simple and reliable method, which does not require purification of recombinant antigen before the serological assay. An hyperimmune guinea pig antiserum produced against purified 6xHis-3ABC was used as competitor in the test. The potential use of this cELISA for the identification of antibodies induced by FMD virus infection from those induced by vaccination is discussed.

Animals↗
PubMed2005

Detection of antibody to the foot-and-mouth disease virus (FMDV) non-structural polyprotein 3ABC in sheep by ELISA.

The specificity and sensitivity of an ELISA for detecting IgG to the 3ABC non-structural protein of foot-and-mouth disease (FMD) virus was evaluated in FMD naive, aerosol-infected, aerosol plus direct contact infected and field-exposed sheep. All 12 sheep that were experimentally infected without prior vaccination seroconverted in the test, although fewer field sera from FMD-exposed sheep were scored seropositive compared to test results for structural protein antibodies. The 3ABC test specificity was 98 or 100% according to whether sera reacting in the doubtful range were scored as positive or negative. The test was then used to investigate the antibody response of sheep vaccinated against FMD and exposed to the virus by an aerosol challenge 4-14 days later. The response of individual animals varied. Whether immunised with high or low doses of vaccine, the development of 3ABC antibody was most likely in sheep from which live virus was recovered at or beyond 9 days post-challenge. Non-structural responses were also more frequent in animals from which multiple incidences of live FMD virus isolation (perhaps more indicative of true virus replication) were demonstrated.

Animals↗
PubMed2005

Utility of recombinant integrin alpha v beta6 as a capture reagent in immunoassays for the diagnosis of foot-and-mouth disease.

Recombinant integrin alpha v beta6 was evaluated as a capture ligand in a sandwich ELISA for the detection and serotyping of foot-and-mouth disease (FMD) virus. Our routinely applied method employs seven serotype-specific rabbit polyclonal antibodies as capture ligands and seven serotype-specific guinea pig polyclonal antibodies as detecting reagents. The recombinant integrin bound FMD virus of all seven serotypes but not that of another vesicular disease, swine vesicular disease (SVD). Considerable heterotypic cross-reactions were evident when using the integrin capture ligand in combination with guinea pig detecting antibodies but totally type-specific reactions resulted when serotype-specific monoclonal antibodies (mabs) were used instead of the guinea pig reagents. The specificity of reaction of the integrin capture/mab detector combination was superior to that of our routinely employed rabbit/guinea pig ELISA and offers an improvement for test interpretation. As a universal trapping reagent for all FMD virus serotypes the alpha v beta6 recombinant protein also has the potential for application in other test procedures for viral identification (e.g. pen-side chromatographic strip-tests, biosensors, immunocapture RT-PCR, antigenic characterization procedures and monoclonal antibody profiling of emerging field virus strains) and in antibody detection assays employed for the diagnosis of FMD.

Animals↗
PubMed2006

Construction and immunogenicity of a recombinant fowlpox virus containing the capsid and 3C protease coding regions of foot-and-mouth disease virus.

Foot-and-mouth disease virus (FMDV) is an important pathogen with worldwide economic consequences. Consequently, an important goal is the development of a vaccine that can provide rapid protection while overcoming the potential risk associated with the production of conventional inactivated vaccines. An important secondary feature of the vaccine would be the ability to distinguish vaccinated from infected animals. A recombinant fowlpox virus (vUTAL3CP1) containing FMDV capsid polypeptide and 3C coding regions of O/NY00 was constructed and evaluated for its ability to induce humoral and cellular responses in mice and guinea pigs. In addition, the ability to protect guinea pigs against homologous virus challenge was examined. Mice and guinea pigs were given booster vaccinations twice and once, respectively, and guinea pigs were challenged 20 days after the booster vaccination. Control groups included animals inoculated with commercial vaccine, fowlpox virus or phosphate-buffered saline (PBS). All animals vaccinated with vUTAL3CP1 developed specific anti-FMDV antibody and neutralizing antibody, as well as T lymphocyte proliferation response and CTL cytotoxic activity. Three of four guinea pigs vaccinated with vUTAL3CP1 were completely protected from viral challenge. The results demonstrated the potential of a fowlpox virus-based recombinant FMD vaccine.

3C Viral Proteases↗
PubMed2006

Relevance of skip metastases for squamous cell carcinoma of the oral tongue and the floor of the mouth.

OBJECTIVE: To analyze the therapeutic implications of the distribution of neck metastases (NM) in patients with squamous cell carcinoma (SCC) of the tongue and the floor of the mouth (FOM). PATIENTS AND METHODS: From January 1987 through December 1997, 339 previously untreated patients with T1-2 N0 M0 SCC of the tongue and the FOM underwent primary surgical treatment in our institution. A retrospective review of the pathology reports and outcome of these patients was made to ascertain the prevalence and distribution of NM. Patients were grouped by clinical neck status at the time of neck dissection: elective neck dissection (END) in the NO neck and subsequent therapeutic dissection (STD) in the neck observed which converted clinically to N+ or regional recurrences after END. All patients were classified according to the American Joint Committee on Cancer (AJCC)/UICC 2002 TNM classification. RESULTS: All patients underwent surgical treatment of the primary cancer and had negative margins at frozen section. Overall incidence of NM was 41.3%. Twenty-seven point eight percent of T1 N0 M0 and 48.2% of T2 N0 M0 patients developed NM (P = .0004). Occult neck metastases occurred in 24.1% of patients. Clinically, N+ metastases occurred in 23.6% of patients. The overall incidence of NM in levels IV and V was 8.5%. Neck level IV nodes were involved in only 1.5% of patients in the END group versus 23.7% in the STD group (P < 0.001). Level V was always associated to nodal metastases in other neck levels. Only 2% of patients in our study presented "skip metastases" in the neck. CONCLUSIONS: Neck levels I and II were at great risk for the development of NM (46.9% and 75.3% respectively). Levels IV (6.5%) and V (2%) were rarely involved in our group of patients. The results found in this study support the indication of supraomohyoid neck dissection for N0 and a more comprehensive neck dissection (levels I-V) for N+ patients in Stage I-II SCC of the tongue and FOM. EBM RATING: C-4.

Carcinoma, Squamous Cell↗
PubMed2004

Participatory diagnosis of a heat-intolerance syndrome in cattle in Tanzania and association with foot-and-mouth disease.

A heat-intolerance (HI) syndrome in cattle in Tanzania was suspected to be associated with previous, clinical foot-and-mouth disease (FMD). A participatory appraisal (PA) method called "matrix scoring" was used to explore livestock-keeper perceptions of association between HI and cattle diseases. A PA method called 'proportional piling' was used to estimate herd incidence of FMD and other diseases, herd incidence of HI, and association between HI and other cattle diseases. Use of matrix scoring and proportional piling with pastoral Maasai informants demonstrated association between FMD and HI. With agropastoral Sukuma informants, the matrix-scoring method did not indicate an association between FMD and HI, whereas the proportional piling method indicated a weak association. Results were supported by calculation of positive predictive values for herder diagnosis of HI and FMD. Clinical examination of cattle by veterinarians was used to confirm HI cases and detection of antibody to non-structural proteins of FMD virus was used to confirm previous clinical FMD.

Animals↗
PubMed2006

Spatial distribution of foot-and-mouth disease in Pakistan estimated using imperfect data.

We estimated the spatial distribution of foot-and-mouth disease (FMD) in Pakistan; we used a probability co-kriging model and the number of FMD outbreaks reported between 1996 and 2000 by Pakistan to the Office International des Epizooties. We used a k-Bessel model and small-ruminant and human densities as surrogate covariates for the population at risk and for livestock markets and movements, respectively. Compared to no or only one covariate, the co-kriging model with both densities provided the best fit to independently obtained data on the spatial distribution of virus isolations (P=0.57). The estimated probability of an FMD outbreak per 25km(2) cell ranged from 0.017 to 0.812, with the maximum relative probability of 47.8 (0.812/0.017). Areas with the highest relative probability of having an FMD outbreak were located in the Punjab region; this is a major animal-production area located along a traditional international animal-trade route.

Animals↗
PubMed2006

Reporting of suspect cases of foot-and-mouth-disease during the 2001 epidemic in the UK, and the herd sensitivity and herd specificity of clinical diagnosis.

We described the clinical diagnostic process utilized during the 2001 epidemic of foot-and-mouth-disease in the United Kingdom (UK), and considered it as a series of diagnostic tests. Premises were classified according to these diagnostic-test results and actual disease status, determined by the reference test, which in this case was one or more internationally accepted laboratory tests. The herd-level sensitivity (HSe) and herd-level specificity (HSp) of the clinical diagnostic process were calculated directly, relative to these internationally accepted reference tests. In this process, the first diagnostic test was 'routine monitoring', which resulted in the identification of suspect cases based solely on the clinical observations of farmers or veterinarians. 6762 suspect cases were identified, and the test had a HSe of 97.6% (95% C.I.: 96.7, 98.3) and a HSp of 95.2% (95% C.I.: 95.0, 95.3). Suspect cases were then subject to the second diagnostic test, termed 'declaration', which consisted of a review of a description of the clinical signs by government veterinarians. Premises that tested positive became 'clinical cases'. The HSe of this test was 97.1% (95% C.I.: 96.2, 97.9), and the HSp was 90.9% (95% C.I.: 90.1, 91.6). During the epidemic, these tests were combined and applied in series, with an overall HSe of 94.7% (95% C.I.: 93.5, 95.7) and an overall HSp of 99.6% (95% C.I.: 99.5, 99.6). We also examined the effect of a policy shift that prohibited delaying the diagnosis pending laboratory testing where the animals exhibited equivocal clinical signs.

Animals↗
PubMed2004

Emergence of a novel subgroup within the widely circulating lineage of foot-and-mouth disease virus serotype Asia 1 in India.

The complete VP1 encoding (1D) gene of 54 foot-and-mouth disease (FMD) virus serotype Asia1 field isolates, most of which were isolated during 2000 and 2001, was sequenced. The phylogenetic analysis identified a novel subgroup (>10% nucleotide divergence) within the widely circulating lineage of this serotype. The newly emerged viruses were responsible for disease outbreaks in both cattle and buffaloes and were present in six different states in the country. Amino acid sequence comparison of these isolates revealed significant sequence divergence at many of the amino acid positions in comparison to those of lineage VI-A and C. Emergence of such viruses may affect the efficacy of vaccine strain currently used for protection against FMD in India.

Amino Acid Sequence↗