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Infectious foot-and-mouth disease virus derived from a cloned full-length cDNA.

A full-length cDNA plasmid of foot-and-mouth disease virus has been constructed. RNA synthesized in vitro by means of a bacteriophage SP6 promoter inserted in front of the cDNA led to the production of infectious particles upon transfection of BHK-21 cells. These particles were also found to be highly infectious for primary bovine kidney cells as well as for baby mice. The difficulty in cloning the foot-and-mouth disease virus cytidyl tract in Escherichia coli was circumvented by joining two separate cloned parts, representing the S and L fragments of the genome, and, in a second step, inserting a dC-dG homopolymer. Homopolymeric sequences of up to 25 cytidyl residues did not lead to the production of virus. Replicons containing poly(C) tracts long enough to permit virus replication were first established in yeast cells. One of these constructs could also be maintained in E. coli and was used to produce infectious RNA in vitro. The length of the poly(C) sequence in this cDNA plasmid was 32 nucleotides. However, the poly(C) tracts of two recombinant viruses found in transfected BHK-21 cells were 60 and 80 nucleotides long, respectively. Possible mechanisms leading to the enlargement of the poly(C) tract during virus replication are discussed.

Animals↗

Interaction of a cellular 57-kilodalton protein with the internal translation initiation site of foot-and-mouth disease virus.

A cellular 57-kDa protein (p57) that binds specifically to the internal translation initiation site in the 5' untranslated region of foot-and-mouth disease virus RNA was detected in cell extracts of different mammalian species by UV cross-linking. The protein binds to two distinct sites of the translation control region which have as the only common sequence a UUUC motif. The first binding site consists of a conserved hairpin structure, whereas the second binding site contains an essential pyrimidine-rich region without obvious secondary structure. Competition experiments indicate that the complexes with the two binding sites were formed by a single p57 species. The protein binds also to the 5' untranslated region of other picornaviruses. Results from footprint analyses with foot-and-mouth disease RNA suggest the participation of additional cellular factors in the translation initiation complex.

Animals↗

Identification of neutralizing antigenic sites on VP1 and VP2 of type A5 foot-and-mouth disease virus, defined by neutralization-resistant variants.

Five neutralizing monoclonal antibodies (nMAbs) obtained against type A5 Spain-86 foot-and-mouth disease virus were used to generate a series of neutralization-resistant variants. In vitro and in vivo assays showed that the variants were fully refractory to neutralization by the selecting nMAb. On the basis of cross-neutralization and binding assays, two neutralizing antigenic sites have been located on the virus surface; one, located near the C-terminus of VP1, displayed a linear epitope, and the second, located on VP2, displayed two conformational epitopes. Nucleotide sequencing of RNA of the parental and variant capsid protein-coding region P1 has placed the amino acid changes at position 198 of VP1 for the first site and at positions 72 and 79 of VP2 for the related epitopes in the second site. The relative importance of these two sites in the biological properties of foot-and-mouth disease virus is discussed.

Amino Acid Sequence↗

trans complementation by RNA of defective foot-and-mouth disease virus internal ribosome entry site elements.

A region of about 435 bases from the 5' noncoding region of foot-and-mouth disease virus RNA directs internal initiation of protein synthesis. This region, termed the internal ribosome entry site (IRES), is predicted to contain extensive secondary structure. Precise deletion of five predicted secondary structure features has been performed. The mutant IRES elements have been constructed into vectors which express bicistronic mRNAs and assayed within cells. Each of the modified IRES elements was defective in directing internal initiation when assayed alone. However, coexpression of an intact foot-and-mouth disease virus IRES complemented four of these defective elements to an efficiency of up to 80% of wild-type activity. No complementation was observed with the structurally analogous element from encephalomyocarditis virus. The role of RNA-RNA interactions in the function of the picornavirus IRES is discussed.

Amino Acid Sequence↗

Inhibition of experimental asthma in man by a new drug (AH 7725) active when given by mouth.

A new drug, AH 7725, with properties similar to those of disodium cromoglycate (Intal) has the additional feature of being effective when taken by mouth. Six patients with allergic asthma were studied by bronchial challenge tests with antigen. In every one the drug was found to inhibit (either partially or completely) the immediate-type asthmatic response. Late responses were not influenced. The results were significant at the 0.5% level.These findings suggest that AH 7725 given by mouth may be able to inhibit naturally occurring asthmatic attacks in allergic patients. In this case its particular advantage as a prophylactic drug would lie in the treatment of very young asthmatic children and other patients who cannot take drugs by inhalation.

Administration, Oral↗

The preference and importance of bathing, toileting and mouth care habits in older persons.

PURPOSE: The goal of this study was to describe bathing, toileting, and mouth care habits and preferences of older persons, as well as the importance of such habits. DESIGN AND METHODS: Fifty-eight community-dwelling older persons were administered the Self-maintenance Habits And Preferences in Elderly questionnaire (SHAPE) during a one-on-one interview with a research assistant. Preferences were described by frequency distributions and mean importance for each item was calculated. Gender differences in preferences were examined using chi(2) analyses, t tests were used to compare importance levels. RESULTS: Women reported spending a longer time bathing, they were more likely to have privacy while bathing, to use a washcloth, a regular size towel, and less likely to use a large towel than were men. One-third of the women reported using some type of absorbent undergarment, while no men did so. Mouth care was the category that received the highest importance ratings. Although order of importance of items was similar across genders, women assigned higher levels of importance to these items than did men. Importance ratings given to atypical habits, such as unusually long or more than 1 bath/day, were higher than those attributed to more usual habits. IMPLICATIONS: Knowledge of the range and importance of such self-care practices can provide information useful in the development of programs and for individualized care.

Aged↗

Effects of inspiratory and expiratory time and high mouth pressures on calculated DLCO by the single-breath procedure. Results with different methods to measure the effective breath-holding time.

We have studied the influence of several manoeuvres modifying inspiratory and expiratory flow, with and without an external obstruction, on measured single-breath diffusing capacity (DLCO). By this method, we could evaluate the effects of increased inspiration or expiration time, independently of the influence of high (negative or positive) mouth pressures. Ten tests were performed in each of 11 trained normal subjects. The sequence of repeated manoeuvres was as follows: (1) basal; (2) inspiratory obstruction (IO); (3) expiratory obstruction (EO); (4) slow inspiration without obstruction (SI), and (5), slow expiration without obstruction (SE). A second, inverted sequence was then performed. The obstructive manoeuvres were forced by a unidirectional valve with a circular hole of 5 mm in diameter. Mouth pressures were registered. The slow flows were voluntarily controlled to be similar to the corresponding flows during the obstructive manoeuvre (approximately 15% of each vital capacity/second). Determinations of breath-holding time (tBH) were calculated by the methods of Jones-Meade, Ogilvie and the American Epidemiology Standardization Project (ESP), but using the same expiratory sample. We found that (DLCO) obtained with the three methods were statistically different (p less than 0.0001) from each other for all manoeuvres. Regarding the basal manoeuvre, there were statistical differences (p less than 0.01) with SI on the Jones-Meade, with IO on the ESP and with SI and IO on the Ogilvie timing methods. Between IO and SI, statistical differences (p less than 0.01) were observed in the three methods, but there was no statistical difference between SE and EO by any method.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗

Distribution of sucrose around the mouth and its clearance after a sucrose mouthrinse or consumption of three different foods.

The distribution of sucrose in whole saliva and in saliva from seven different regions of the mouth was determined in 10 subjects over the 10-min period following the chewing of a doughnut, sucking on a mint candy, the drinking of orange juice, or use of a 10% sucrose mouthrinse. With all products, the sucrose was distributed non-uniformly, with particularly low concentrations on the lingual surfaces of the lower incisors and the facial surfaces of the upper molars. Clearance was also most rapid from these sites. Since the depth and duration of a Stephan curve in dental plaque is influenced by the sugar concentration to which the plaque is exposed, the results, together with previous results on salivary film velocity in different regions of the mouth, help to provide an explanation for the site-specificity of smooth-surface caries and of supragingival calculus deposition.

Adult↗

Antihypertensive action of medullipin I given by mouth.

Perfusion of normal rat kidneys with 5% human albumin in a balanced salt solution bubbled with oxygen yielded medullipin I (Med I) in the renal venous effluent. The presence of Med I in the renal venous effluent has been established by thin-layer chromatography, by the type of vasodepressor effect when injected intravenously as a bolus into the hypertensive rat, by inhibition of the vasodepressor effect of the renal venous effluent by Tween 20 and SKF 525A (proadifen, inhibitor of cytochrome P-450), and by removal of the liver from the circulation (a procedure that inhibits extracted Med I). Med I so derived lowered blood pressure of spontaneously hypertensive rats when injected into the stomach by an indwelling tube or when given by mouth. The lowering of blood pressure was attended by no change in cardiac output and no change in heart rate. Med I given by mouth to the spontaneously hypertensive rat is a vasodilator that suppresses sympathetic tone, acting in the same way as Med I extracted from renal papillae and given intravenously. Importantly, the antihypertensive action was demonstrated in the spontaneously hypertensive rat, a model of hypertension considered to mimic idiopathic or essential hypertension of humans. Med I is a promising therapeutic agent for hypertension.

Administration, Oral↗

A minimally obtrusive, secure mask for prevention of access to the mouth.

Several childhood neurological conditions can lead to problems with unintended access to the mouth. Earlier workers have demonstrated benefits in the use of a helmet restricting mouth access, within a behavioural modification programme. Conventionally, such headgear is very obtrusive. This paper presents a simple technique using conventional plaster-based moulding, which has been successfully used in making secure, minimally obtrusive masks. Masks produced for two children have been readily accepted and have been successful. Any such headgear should be used only within a managed behavioural modification programme.

Adolescent↗

Differentiation of dry mouth etiology.

Dry mouth is a common complaint. While it is often considered to be synonymous with the diagnosis of salivary gland disease, this is not the case. There are many causes of the symptomatic complaint of oral dryness, both salivary and non-salivary. This paper will review the causes of "dry mouth" and the means of establishing the underlying diagnosis.

Diagnosis, Differential↗

Symphysiotomy, symphysiectomy, and intermandibular arthrodesis in a cat with open-mouth jaw locking--case report and literature review.

A Persian cat was presented with open-mouth jaw locking. It was suspected that abnormal extrusion of the maxillary canine teeth caused contact with the mandibular canine teeth on closure of the mouth, resulting in levering forces that may have led to increased symphyseal mobility, temporomandibular joint laxity, rotational movement of the left mandibular body and locking of the left coronoid process lateral to the zygomatic arch. Symphysiotomy, symphysiectomy, circumferential and interdental wiring, and intraoral resin-based splint application were performed to achieve intermandibular arthrodesis. Following removal of all devices, no further jaw locking episodes were reported during a 33-month follow-up period.

Animals↗

Use of in situ hybridization for the detection of foot-and-mouth disease virus in cell culture.

Biotinylated complementary DNA (cDNA) and RNA probes were prepared from a specific and highly conserved section of the foot-and-mouth disease virus (FMDV) genome coding for the RNA-dependent RNA polymerase. Hybridization was conducted on FMDV-infected, bovine enterovirus (BEV)-infected, and noninfected swine kidney cell cultures. The detection system utilized the enzyme system streptavidin-alkaline phosphatase, the substrate phosphate, and the chromogen nitroblue tetrazolium. Intense cytoplasmic granular staining was present at 2 and 4 hr postinfection (hpi), with less staining observed at 24 hpi. The staining was specific for FMDV, as indicated by a lack of staining of noninfected cells and BEV-infected cells. With the RNA probe, positive cells were detected up to the highest viral dilution assayed, which was approximately 96 TCID50. The cDNA probe was slightly less sensitive, detecting positive cells at 10-fold lower dilutions. This technique could prove useful in the diagnosis of foot-and-mouth disease in animals or in the detection of FMDV in biologics submitted for importation.

Animals↗

Fluconazole sensitivities of Candida species isolated from the mouths of terminally ill cancer patients.

Oral candidosis is common in advanced cancer and is often treated with the systemic triazole antifungal drug fluconazole. This study examined the species of yeast present in the mouths of 30 patients with advanced cancer and determined their sensitivity to fluconazole. Thirty-five yeast isolates were collected from a total of 25 (83 percent) of the patients sampled. The two most common species were Candida albicans (15 isolates) and C. glabrata (11 isolates)--with smaller numbers of C. tropicalis, C. parapsilosis, C. guilliermondii, C. inconspicua, and Saccharomyces cerevisiae. The minimal inhibitory concentrations (MIC) of fluconazole for the strains of C. albicans were generally low (median 0.19 microgram/ml) but were considerably higher for C. glabrata (median 2 micrograms/ml). The remaining species demonstrated MICs similar to those for C. albicans, with the exceptions of C. inconspicua and Saccharomyces cerevisiae, which were relatively insensitive. In conclusion, non-albicans yeasts are common in the mouths of patients with advanced cancer and these may have reduced sensitivity to fluconazole. Mycological diagnosis is a valuable aid to management.

Adult↗

Mouth occlusion pressure, CO2 response and hypercapnia in severe chronic obstructive pulmonary disease.

The resting mouth occlusion pressure 0.1 s after onset of inspiration (P0.1) and minute ventilation (V'E) and their response to CO2 in patients with chronic obstructive pulmonary disease (COPD) remain controversial. The ventilatory drive and the factors that predict resting arterial CO2 tension (Pa,CO2) were studied in 19 eucapnic and 14 hypercapnic severe COPD patients, and 20 controls. The CO2 response was evaluated by the Read technique. The V'E, and P0.1 as a function of end-tidal CO2 tension (Pet,CO2) was used to study the ventilatory (deltaV'E/deltaPet,CO2) and P0.1 response (deltaP0.1/deltaPet,CO2). In the patients, respiratory muscle function and pleural occlusion pressure 0.1 s after onset of inspiration (Ppl,0.1) were evaluated with simultaneous measurement of pleural (Ppl) and gastric (Pga) pressures. Hypercapnic patients had lower forced vital capacity (FVC), forced expiratory volume in one second (FEV1), and arterial O2 tension (Pa,O2). Resting P0.1 was higher in patients than in controls, whereas deltaP0.1/deltaPet,CO2 was similar in the three groups. There was no difference in resting P0.1 (3.6+/-2.0 versus 4.3+/-2.8 kPa (2.7+/-1.5 versus 3.2+/-2.1 cmH2O), p=0.2) and Ppl,0.1 (1.4+/-2.3 versus 5.2+/-3.3 kPa (4.08+/-1.7 versus 3.9+/-2.5 cmH2O), p=0.22) between eucapnic and hypercapnic COPD, whereas deltaV'E/deltaPet,CO2 was lower in the hypercapnic group (0.29+/-0.24 versus 0.66+/-0.5 L x min(-1) x kPa, p<0.001). By logistic regression only FEVI and increased diaphragmatic load, and not respiratory drive, predicted resting Pa,CO2. Irrespective of CO2 level, baseline central drive (represented by the mouth occlusion and pleural pressures) and CO2 response are preserved in most patients with severe chronic obstructive pulmonary disease. Effective ventilation is inadequate in the more severely obstructed patients and this results in hypercapnia. Neuroventilatory coupling failure is an attractive explanation for chronic hypercapnia in these patients.

Aged↗

New compressed tablet rapidly disintegrating in saliva in the mouth using crystalline cellulose and a disintegrant.

We formulated a new compressed tablet which is rapidly disintegrated and dissolved in the mouth without the need to drink water. The crushing strength increased with increasing compression force and maximum values (8-18 kgf) for the formulations comprising crystalline cellulose and low-substituted hydroxypropylcellulose (L-HPC) were obtained at the compression force of 300 kgf. Rapid disintegration (within 30 s) was obtained in vitro using various compounding ratios of crystalline cellulose to L-HPC. Tablets prepared with crystalline cellulose and L-HPC rapidly disintegrated in saliva (small amount of water) in the mouth of humans.

Cellulose↗

Estimation of 140S particles in foot-and-mouth disease virus (FMDV) vaccine by using the computer analyzing system.

The quantity of 140S particles in inactivated foot-and-mouth disease virus (FMDV) vaccine samples produced in Foot-and-Mouth Disease Vaccine Production Center (FMD Vaccine Production Center) in Thailand was estimated by the sucrose gradient ultracentrifugation and optical density analysis by using the computer applying system. The soft ware; Chromato Data System (CDS) (Nihon Chromato Works Co., Ltd. Japan) which is prepared for the analysis of chromatography, was applied for the estimation of 140S particles in FMDV vaccine. The quantity of 140S particles in each vaccine sample measured by CDS was mostly ranged from 2-4 micrograms/ml and this quantity was consistent with the results of the other reports. This method is considered to be the available method for estimation of 140S particles in FMDV vaccine as routine assay.

Aphthovirus↗

Reduction of self-injurious hand mouthing using response blocking.

We evaluated a response-blocking procedure for reducing the self-injurious hand mouthing of 2 adults with profound disabilities. The procedure reduced mouthing in both cases. The results suggest research is warranted to delineate the behavioral process responsible for the effect of response blocking and to determine the procedure's long-term utility.

Adult↗