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PubMed2006

Multiplexed detection of antibodies to nonstructural proteins of foot-and-mouth disease virus.

Liquid array technology was used to develop a multiplexed assay for the detection of antibodies to viral nonstructural proteins (NSPs), raised in cattle in response to infection with foot-and-mouth disease (FMD) virus. Two assays, one based on recombinant NSPs and the other on synthetically produced peptides, were developed and compared side-by-side. Serum samples from serial bleeds of cattle, each experimentally infected with one of the seven serotypes (C, A, O, Asia, SAT1, SAT2, SAT3) of FMD virus were analyzed. A distinct pattern in the detection of NSP antibodies and a close correlation of the recombinant protein and peptide-based assays were observed. The detection of antibodies to NSPs is a method to differentiate FMD-infected and FMD-vaccinated animals, and a high-throughput assay would be an invaluable tool in the case of an outbreak of FMD in North America, when emergency vaccination may be utilized to spare vaccinated, noninfected animals from slaughter and subsequent disposal.

Animals↗
PubMed2001

Molecular epidemiology of serotype O foot-and-mouth disease virus with emphasis on West and South Africa.

Genetic relationships of serotype O foot-and-mouth disease (FMD) viruses recovered from outbreaks of the disease in the West African countries of Niger, Burkina Faso and, Ghana (1988-1993) and those from South Africa (2000) were determined by partial VP1 gene characterization. A 581-bp fragment, corresponding to the C-terminus half of the ID (VP1 gene) region was amplified and sequenced. An homologous region of 495 nucleotides was ultimately used to determine genetic relationships of serotype O viruses from the Middle East, Europe, South America, North Africa, East Africa, southern Africa and Asia. Seven distinct type O genotypes were identified by phylogenetic reconstruction, consisting of viruses from the following geographical regions: Genotype A: Asia, the Middle East, and South Africa, Genotype B: East Africa, Genotype C: West and North Africa, Genotype D: Taiwan and Russia, Genotype E: Angola and Venezuela, Genotype F: Western Europe, and Genotype G: Europe and South America. The genotypes constitute three different evolutionary lineages (I-III), which correspond to three discrete continental regions, some of which display inter-continental distributions due to introductions. Results further indicate that the outbreaks in Burkina Faso (1992) and Ghana (1993) are part of the same epizootic and that the strain involved in a recent outbreak of the disease in South Africa is most closely related (97% sequence identity) to a 1997 Bangladesh strain.

Africa, Western↗
PubMed2002

The complete nucleotide sequence of the PanAsia strain of foot-and-mouth disease virus isolated in Japan.

The complete nucleotide sequence of the foot-and-mouth disease virus (FMDV) O/JPN/2000 strain, the PanAsia strain, was determined by cycle sequencing and primer walking. The 5' end of the genome upstream from homopolymeric poly(C) tract (S-fragment) was 367 nucleotides in length, and the remainder of the genome (L-fragment), excepting the poly(A) tail, was 7808 nucleotides. The L-fragment contains a single open reading frame of 6996 nucleotides terminating at a UAA codon 96 bases from the 3' poly(A) sequence. Comparison of sequences shows that the length of the structural and non-structural protein coding regions are identical to those in the O1/Kaufbeuren strain, and no striking differences such as deletion or insertion were observed between them.

Animals↗
PubMed2003

Use of the reverse transcription polymerase chain reaction (RT-PCR) for the rapid diagnosis of foot and mouth disease in South America.

Foot and mouth disease (FMD) is a limiting factor for the economic progress of the animal industry in South America. The presence of the disease results in the imposition of national and international sanitary barriers to animals and animal products, and, most especially, a reduction in the availability of protein from animal origin and in income. Rapid and accurate identification of infected animals, those with either clinical or subclinical disease as well as with persistent infection, is essential for maintaining an efficient eradication programme. The polymerase chain reaction was used to rapidly identify infected animals. With a primer set that corresponds to a conserved region of the 3D sequence of the viral genome, it was possible to amplify, regardless of the serotype, 116 strains of FMD virus, of which 109 were strains collected from outbreaks of FMD throughout South America from 1945 to the most recent outbreaks in 2000/2001. The PCR technique should be of considerable value in facilitating the diagnosis of FMD in South America. where laboratory resources are limited and a rapid response is needed, particularly in areas where national programmes for controlling or eradicating the disease are being implemented.

Animals↗
PubMed2003

Molecular epidemiology of SAT3-type foot-and-mouth disease.

VP1 gene nucleotide sequences of 51 SAT3-type foot-and-mouth disease (FMD) viruses from seven southern and eastern African countries were used to infer a gene phylogeny. Results obtained by phylogenetic analysis of the homologous 405 nt region corresponding to the C-terminal 128 amino acids of 1D and adjacent 7 amino acids of 2A indicate that there are six distinct virus lineages evolving independently in different geographical localities in accordance with the FMD topotype concept. Topotypes I-IV occur in southern Africa, whilst topotypes V and VI are unique to East Africa. Viruses of different topotypes differ from each other at 20% or more of the nucleotide sites, specified in this study. Despite the limited geographical distribution of this serotype, the level of intratypic variation is intermediate between that of SAT1 and SAT2, both of which are widely distributed in sub-Saharan Africa. Within SAT3, 37.3% and 47.4% of sites were completely conserved on nucleotide and amino acid levels, respectively. The locality-specific grouping of viruses permits accurate determination of the sources of outbreaks, whilst the high levels of variation within the immunodominant 1D protein has implications for the control of the disease through vaccination.

Africa, Eastern↗
PubMed2004

Detection of foot and mouth disease virus by RT-PCR and microplate hydridization assay using inactivated viral antigens.

A single step RT-PCR was tested for detection of foot and mouth disease virus (FMDV) and immunoenzymatic determination of amplified products in a microplate hybridization assay. Inactivated reference strains (ELISA antigen) of all seven serotypes were used to optimize the test. Oligonucleotide primers were selected from two different genomic regions coding for RNA polymerase and VP1 protein, respectively. The RT-PCR used to amplify the polymerase gene specific RNA detected FMDV strains A, C, O, Asial and SAT1, and the identity of the fragments obtained was confirmed with a specific internal biotin-labelled capture probe. For the amplification of the VP1 genome region, two sets of oligonucleotide primers were used. One primer pair was successfully applied for the detection of serotypes A, C, O and Asial and a second one for serotypes SAT1, SAT2, SAT3. The specific probe enabled the detection of all the amplified products in a PCR ELISA test. By comparison with antigen ELISA, the PCR ELISA method allowed the detection of smaller amounts of FMDV in the inactivated material examined. The application of molecular diagnostic methods to inactivated antigens offers a good alternative procedure for developing and optimizing a sensitive method for detection of FMDV in laboratories that are not allowed to work with viable FMDV.

Animals↗
PubMed2004

Comparison and analysis of the complete nucleotide sequence of foot-and-mouth disease viruses from animals in Korea and other PanAsia strains.

During the last 3 years, foot-and-mouth disease virus serotype O, named PanAsia, caused two outbreaks in the Republic of Korea. To determine if there was an obvious genetic relationship between the virus isolated in 2002 (O/SKR/2002) and the O/SKR/2000, and to further analyze the epidemiological relationships between the PanAsia viruses and the viruses identified in Korea, the complete nucleotide sequence of the O/SKR/2002 and the O/SKR/2000 were determined by automatic cycling sequencing and primer walking. The nucleotides and the deduced amino acid (aa) sequences of the strains identified in Korea were compared with each other and also those enrolled in the GenBank database. In comparison and analysis of the viruses identified in Korea, any deletions or insertions in the specific fragment gene of both the O/SKR/2002 and O/SKR/2000 were not identified. However, comparison of the aa sequence of the identified virus in 2002 from pigs with those of other PanAsia strains revealed significant substitutions of 4 aa in the VPI region and 8 aa in the 3A region. In phylogenetic analysis based on the translated region, the identified virus in 2002 appeared to be the divergence of approximately 1% degree with other PanAsia viruses. Also, animal experiments indicated that O/SKR/2000 is not host-restricted and develop the clinical signs in the main susceptible livestock species (cattle and pigs). However, O/SKR/2002 did not develop the clinical signs in cattle and showed severe clinical signs only in pigs. These analytic data suggest that 2002 outbreaks in Korea is not re-occurred but re-introduced from nowhere.

Amino Acid Sequence↗
PubMed2004

[Burning mouth syndrome--etiology, differentialdiagnostical aspects and therapy].

Burning Mouth Syndrome (BMS) or glossodynia is a chronic pain syndrome that mainly affects middle-aged/elderly women. This condition is probably of multifactorial origin, often idiopathic, and its etiopathogenesis remains largely enigmatic. Visible pathologic lesions of the oral mucosa or processes are usually not evident. Etiologic factors that have been reported include several disorders (e. g. hematologic disorders, denture factors, the climacteric, infections, endocrinological, neurological or psychiatric disorders). On the other hand psychological factors, such as anxiety, depression and phobias have been reported to play a significant role at the beginning of this nosological entity. BMS related to nutritional deficiency is uncommon. A systemic approach to assessment is essential and the opinion of other specialities, particularly internal medicine, dentistry or neurology, may be of help. Treatment is of the underlying cause but in many cases no specific aetiological factor is identified. Non-indicated substitution therapy of suspected but unproved deficiency states must be avoided. Treatment approaches were divided into topical and systemic strategies. However, there is little research evidence that provides clear guidance for those treating patients with BMS.

Burning Mouth Syndrome↗
PubMed2001

Helicobacter pylori colonization of tongue mucosa--increased incidence in atrophic glossitis and burning mouth syndrome (BMS).

Nested polymerase chain reaction (PCR) was performed to detect the presence of Helicobacter pylori in tongue mucosa in 268 patients divided into four groups according to their diagnosis: 87 with atrophic glossitis, 37 with benign migratory glossitis and 144 with burning mouth syndrome (BMS). The latter group was subdivided according to anatomic site of burning sensation: subgroup A (54 patients) with complaints limited to tongue and subgroup B (90 patients) with burning sensations in other parts of oral mucosa. H. pylori was found in 43 samples (16%). Bacteria were significantly less present in tongue mucosa affected with benign migratory glossitis compared with atrophic glossitis and BMS (P=0.025). This difference was more obvious when compared with atrophic glossitis only (P=0.006). Mucosal changes in these conditions might make the oral environment more acceptable for H. pylori colonization compared with normal mucosa, and this mechanism may play a role in its oro-oral transmission.

Burning Mouth Syndrome↗
PubMed2002

Macrophage phagocytosis of foot-and-mouth disease virus may create infectious carriers.

Macrophages play critical roles in innate defences against virus infections, particularly pertinent to the rapid immune response required following emergency vaccination against foot-and-mouth disease virus (FMDV). Consequently, macrophage-FMDV interaction was studied in vitro, in the absence of specific antibodies, to mimic the animal early postvaccination. A gradual loss of infectivity and viral antigen was observed over 48 hr, and no evidence of productive virus replication was found. From the pathological viewpoint, an important observation was that the majority of macrophages carried infectious virus for at least 10 hr. Pronase and mild acid treatments showed the virus to be primarily on the cell surface during the first 4 hr. Thereafter, it became internalized (pronase- and pH resistant), but remained infectious for 10-24 hr. The internalization process was dependent on microfilament activity, while the survival of infectious virus related to live virus-dependent inhibition of macrophage protein synthesis. Infectious centre assays demonstrated that this infectious virus - whether on the cell surface or internalized - was actually being released from the cells. This is interesting considering that FMDV is highly pH labile. Together, these characteristics suggest that the virus had been internalized by a process such as macropinocytosis, and fusion with endosomes was delayed or impaired. This mechanism whereby the virus could 'piggyback' on or in the macrophage, becoming internalized but not degraded for at least 10 hr, are important considerations in FMD pathogenesis. Such 'virus-transporting' macrophages would be in a position to carry infectious FMDV to different sites in the body, where it could be released to infect other cells for replication.

Actin Cytoskeleton↗
PubMedDate not supplied

Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis.

A reverse transcription-PCR (RT-PCR) method is presented for the highly sensitive and specific detection of foot-and-mouth disease virus (FMDV). A primer pair flanking a region of the viral polymerase gene (3D) corresponding to the C-terminus of the protein was designed and a single step RT-PCR reaction was developed. The assay allowed the detection of viral RNA from a variety of animal samples and from a wide range of FMDV isolates of different origins and serotypes. The presence of an Ahd I restriction site within the amplicon in 96% of the isolates analyzed allowed an additional confirmation step of the positive reactions by a simple digestion yielding characteristic fragment sizes. The set of primers described here was suitable for direct sequencing of the PCR product (290 bp), and the nucleotide sequences corresponding to the SAT 1 and SAT 3 strains were determined. The segment amplified, when used in phylogenetic studies, allowed the clustering of SAT isolates and the rest of FMDV strains as two separate lineages.

Animals↗
PubMedDate not supplied

Utility of automated real-time RT-PCR for the detection of foot-and-mouth disease virus excreted in milk.

Foot-and-mouth disease virus (FMDV) can be excreted in milk and thereby spread infection to susceptible animals in other holdings. The feasibility of using real-time reverse transcription polymerase chain reaction (rRT-PCR) as a diagnostic tool for detection of FMDV in milk was assessed by studying the excretion of virus from experimentally-infected cattle. Fore- and machine milk samples were collected over a 4-week period from two dairy cows infected with FMDV and from two in-contact cows held in the same pen. The whole, skim, cream and cellular debris components of the milks were tested by automated rRT-PCR and results compared to virus isolation (VI) in cell culture. The onset of clinical signs of FMD in all four cows correlated with viraemia, and the presence of FMDV in other clinical samples. rRT-PCR results matched closely with VI in detecting FMDV in all milk components and generally coincided with, but did not consistently precede, the onset of clinical signs. rRT-PCR detected FMDV in milk up to 23 days post inoculation which was longer than VI. Furthermore, the detection limit of FMDV in milk was greater by rRT-PCR than VI and, in contrast to VI, rRT-PCR detected virus genome following heat treatment that simulated pasteurisation. rRT-PCR was also able to detect FMDV in preservative-treated milk. In conclusion, this study showed that automated rRT-PCR is quicker and more sensitive than VI and can be used to detect FMDV in whole milk as well as milk fractions from infected animals.

Animals↗
PubMed2006

Quantification of within- and between-pen transmission of Foot-and-Mouth disease virus in pigs.

Quantified transmission parameters of Foot-and-Mouth Disease Virus (FMDV) are needed for epidemic models used for control and surveillance. In this study, we quantified the within- and between-pen transmission of FMDV in groups of pigs by estimating the daily transmission rate beta, i.e. the number of secondary infections caused by one infectious pig during one day, using an SIR (susceptible-infectious-removed) model. Within-pen transmission was studied in four groups of ten pigs in which 5 infected and 5 susceptible pigs had direct contact; between-pen transmission was studied in one group of ten pigs in which 5 infected and 5 susceptible pigs had indirect contact. Daily results of virus isolation of oropharyngeal fluid were used to quantify the transmission rate beta, using Generalised Linear Modelling (GLM) and a maximum likelihood method. In addition, we estimated the expected time to infection of the first pig within a pen T(w) and in the indirect-contact pen T(b). The between-pen transmission rate beta(b) was estimated to be 0.59 (0.083-4.18) per day, which was significantly lower than the within-pen transmission rate beta(w) of 6.14 (3.75-10.06). T(w) was 1.6 h, and T(b) was 16 h. Our results show that the transmission rate is influenced by contact structure between pigs.

Animal Husbandry↗
PubMed2002

Dose-dependent responses of sheep inoculated intranasally with a type O foot-and-mouth disease virus.

Unlike foot-and-mouth disease (FMD) in cattle and pigs, which spreads rapidly, resulting in easily detectable foci of clinical infection, the disease in sheep is characterized by restricted transmission, low morbidity and sporadic clinical cases. The study described was designed to investigate whether the ability of sheep to transmit and maintain FMD virus was dose-related. The viral isolate used was known to be associated epidemiologically with rapid fade-out of transmission within sheep flocks. Five separate transmission experiments were performed, with different doses of FMD virus, each experiment containing five intranasally inoculated donor sheep and 10 in-contact recipient sheep. The lowest dose required to cause clinical infection by inoculation (10(4) 50% tissue culture infectious doses; 10(4) TCID50) was also the optimum dose for producing in-contact transmission. Inoculation of donor sheep with higher doses (10(5) and 10(6) TCID50) resulted in reduced transmission, characterized by reduced duration and degree of viraemia and an early humoral and cell-mediated immune response. Principal component analysis was used to interpret the complex interactions of the dose-related responses to infection.

Administration, Intranasal↗
PubMedDate not supplied

[Epitheloid leiomyosarcoma of the mouth in childhood].

A six-year-old girl is presented with an increasing mass involving the anterior vestibule and the floor of the mouth. Histologic and immunohistochemical examination revealed a poorly differentiated epitheloid leiomyosarcoma with destruction of the mandible. According to the CWS-96-study the patient underwent preoperative chemotherapy followed by complete resection of the mandibular body with the surrounding soft tissues. Recurrent tumor became evident only three months later on. The further treatment consisted of tumor resection, oral chemotherapy and irradiation. Nevertheless tumor control could not be achieved. The patient died of progressive disease 16 months after diagnosis. Leiomyosarcoma is extremely uncommon in childhood, especially with localisation in the oral cavity. Diagnosis is based on histologic examination and immunohistochemistry. In the presented case the lack of smooth muscle actin expression made diagnosis difficult. The preoperative chemotherapy could not achieve reduction of tumor size, so that extensive surgery became necessary. Estimation of the prognosis of the oral leiomyosarcomas in childhood is difficult. High grade tumours and involvement of bone seem to be associated with bad clinical outcome. The presented case is confirming that.

Adolescent↗
PubMed1992

[Treatment and treatment results of mouth floor cancers].

The records of 68 patients with cancer of the floor of the mouth were reviewed. 56 patients underwent surgical management, 51 of them got additionally postoperative radiation. The tumour-specific five years' survival for patients with operation was 46%, 57% of treatment failures developed from local recurrence of the tumour. In 52% of all cases there was a spread to the lymphatic system in the histological evaluation. There was high incidence of false negative clinical examinations of the neck. Conservative neck dissection was the procedure of choice for clinically positive lymph nodes and for the elective management of the neck. Only advanced tumours showed involvement of the mandibular bone. Therefore a conservative management of mandibular resection was preferred. Radical tumour extirpation and histological controlling with serial sectioned specimens are methods of avoiding local tumour recurrence.

Carcinoma, Squamous Cell↗
PubMed1995

[The lymph vessel system of the mouth cavity and pharynx].

BACKGROUND: The understanding of pathological processes involving the lymphatic system of the upper digestive tract requires a detailed knowledge of the lymphovascular architecture of this region. METHODS: The lymphatic system of the upper digestive tract was examined in 390 patients by different methods such as light microscopy, electron microscopy, histochemistry, and indirect lymphography. RESULTS: The mucous membrane of the upper digestive tract is permeated by two communicating lymphatic networks with regionally varying density. There are no areas without lymphatics. The lymphatic fluid of the nasopharynx flows from the posterior wall mainly towards the retropharyngeal and accessory lymph nodes (level II). The lymphatic flow from the buccal mucous membrane, the anterior floor of the mouth, the oral tongue, and the hard palate mostly drains toward the submandibular and craniojugular areas. Lymphatic fluid from the tonsils, the soft palate, and the hypopharynx flows primarily to the deep cervical lymph nodes (level II and III). CONCLUSION: These results are a basis for the understanding of a) secondary lymphatic edemas and b) frequency and direction of lymphogenous metastasis of carcinomas in this region. The direction of metastasis usually follows the physiologic lymphatic drainage. Metastasis towards untypical areas can in most cases be explained by an alteration of the lymphatic flow secondary to inflammation, radiotherapy, previous surgery, and subsequent scar formation or tumor growth.

Humans↗
PubMed1993

[CT and MRI in tumors of the mouth and oropharynx--comparison of methods with reference to fast and ultra-fast MR pulse sequences].

A total of 488 patients with neoplasms (in 474 cases with malignant tumours) of the oropharynx, floor of the mouth and oral cavity, were examined with MRI and CT. MRI proved superior to CT, on account of its coronal and sagittal slice orientation enabling not only a better demonstration of findings, but also improved tumour detection and staging. The diagnostic value of MRI was further enhanced with additional fast and ultrafast MR sequences rather than spin echo sequences alone, as previously practised.

Adenocarcinoma↗