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Biomedical subjects

D M Moore

Publications and source records attributed to D M Moore.

At least 19 recordsLinked to original sources

Technical note: mineral deposits on Dacron bags during ruminal incubation.

Extensive DM contamination was found on Dacron bags that were incubated for prolonged periods of time in the rumen of steers fed alfalfa hay. The ash content of the contaminant was high, and most of it was acid-soluble X-ray analysis indicated the presence of hydroxylapatite and synthetic calcium magnesium phosphate or whitlockite. The contaminant appeared as a smooth coating on the Dacron fiber, suggesting that contamination was a gradual process rather than the result of entrapment of dislodged crystals from plant material. Contamination seemed to occur exponentially within the range of observations (0 to 42 d). Contamination also occurred in steers fed orchardgrass, although to a lesser extent than in steers fed alfalfa hay. The DM contamination was less than .04 g per bag (average bag weight was 1.2 g) during the first 10 d of incubation. However, correction for contamination might be required for studies involving longterm incubation or mineral digestion.

Animal Feed

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

Antigenic stability of foot-and-mouth disease virus variants on serial passage in cell culture.

Two neutralizing monoclonal antibody (MAb)-resistant variants selected from an isolate of foot-and-mouth disease virus (FMDV) type A5 were repeatedly passaged in cell culture and monitored for susceptibility to neutralization by the selecting MAb. A variant isolated with a MAb to a conformational epitope (1-OG2) lost resistance in 20 passages, while a variant isolated with a MAb to a linear epitope (1-HA6) persisted for 30 passages. In both cases, the virus population emerging after passage was antigenically and genetically indistinguishable from the original wild-type parental virus (FMDV A5 Spain-86). Coinfection assays with the wild type and each variant, and between the variants, showed rapid conversion to a homogeneous population. Wild-type virus prevailed over the variants and for coinfection between the variants, the linear epitope variant 1-HA6. While both variants arose from a single nucleotide substitution and reversion to wild type occurred for each, it appears that the variant based on the continuous epitope (1-HA6) was more stable. We discuss the implications of these results for the antigenic diversity of FMDV and its relationship to virus evolution.

Animals

The electrically evoked vestibulo-ocular reflex: I. Normal subjects.

Recent animal studies indicate that electric currents applied through perilymphatic-space electrodes stimulate vestibular primary afferent neurons directly. These findings suggest that electrical stimulation may provide a testing method by which the vestibular nerve and central pathways could be evaluated separately from the vestibular end-organ. The goal of this study was to obtain normative data on human beings for an electrically evoked vestibulo-ocular reflex (EVOR). Sinusoidal electrical stimuli (0.0125 to 0.8 Hz, 4 mA peak intensity) were applied along the interaural axis through mastoid electrodes in 10 subjects. Horizontal eye movements were recorded by an infrared limbus-tracking device. The subjects also underwent rotational stimulation at the same frequencies so that their horizontal vestibulo-ocular reflex (VOR) could be evaluated. Nystagmus was observed in the EVOR at lower stimulus frequencies, whereas purely sinusoidal eye deviations occurred at higher frequencies. The phase of the EVOR slow-component eye velocity consistently lagged the stimulus. This contrasts with the phase measurements of the VOR in the same subjects, which exhibited a lead relative to head velocity. These findings suggest that currents applied to human beings may activate vestibular primary afferents independent of peripheral receptor mechanisms and thereby provide a "site-of-lesion" testing method by which the vestibular nerve and central pathways can be evaluated separately from the vestibular end-organ.

Adolescent

Occult primary tumors. The management of isolated submandibular lymph node metastases.

The management of patients with cervical metastases from unknown primary tumors presents a therapeutic challenge to both the head and neck surgeon and radiotherapist. If after careful search the primary tumor remains truly occult, traditional methods of radiotherapy encompassing fields from the base of skull to clavicles are often employed, with significant attendant morbidity. To determine if more limited therapy would be effective in cases of isolated regional lymph node metastases, the patterns of tumor spread to nodes in the submandibular region were studied. A retrospective analysis of 472 radical neck specimens obtained from 1975 to 1985 revealed 19 cases (4.0%) of cervical metastases limited to the submandibular triangle. Sources of these tumors included lip (3), buccal mucosa (4), nasal vestibule (1), floor of mouth (4), alveolar ridge (3), oral tongue (1), and unknown (3). It appears that solitary submandibular nodal metastases predominantly arise from sites in the oral or nasal cavity. This suggests that in patients with isolated submandibular lymph node metastases from occult primary sites, a more conservative therapeutic approach to potential primary sites is indicated after treatment of the metastatic focus.

Adult

Antibodies elicited by a biosynthetic peptide related to a major immunogenic area of FMDV A12.

Foot-and-mouth disease virus (FMDV) capsid contains 60 copies each of four structural proteins, virus proteins 1-4. Virus protein 1 (VP1) plays an important immunogenic role, being the only VP that is immunogenic as an isolated protein. Even peptides representing a partial amino acid (AA) sequence of VP1 can induce protective immunity in experimental hosts. A 32 AA residue, in a tandem repeat configuration (32dimer), of sero/subtype A-12 Lp ab VP1 (AA 132-168) was highly immunogenic for its homologous subtype and partially protective for FMDV serotype A strain A24. This cross-reactivity was further demonstrable in the ELISA and mouse protection tests. Three different antibody populations were isolated by affinity chromatography (AFC) from the serum of a steer immunized with the 32dimer. Each population seems to recognize a different epitope on the 32dimer peptide since each fraction was defined as unique by its reactivity with different subtypes of FMDV virus in RIA, ELISA, neutralization and competition assays. Considering the neutralizing activity of each of the antibody populations the pattern of neutralization of the 32dimer elicited antiserum can be described. Two of the three epitopes were mapped by competition assays using synthetic peptides.

Animals

Cancer of the tongue base treated by a transpharyngeal approach.

Tongue base resection plays an important role in the management of the patient with cancer of the posterior tongue. The considerable morbidity resulting from loss of functional tongue includes compromised deglutition, chronic aspiration, and altered speech articulation. The particular operative approach used dictates additional secondary morbidity, which may include cosmetic and functional defects of the mandibular arch, malocclusion, lip and chin scars, and postoperative changes in the oral cavity. Our recent experience with a transpharyngeal approach in 13 patients with tongue base cancer is reviewed. Adequate operative exposure was obtained in all cases. Persistent aspiration was not a problem, and all patients learned to swallow effectively. The transpharyngeal approach avoids unnecessary surgical trauma to the mandible and anterior oral cavity and minimizes cosmetic deformity. Our early experience with this approach is encouraging.

Aged

Protection of cattle and swine against foot-and-mouth disease, using biosynthetic peptide vaccines.

A single dose of foot-and-mouth disease (FMD) virus protein 1 (VP1) peptide, expressed in Escherichia coli as a fusion protein with 190 amino acids (AA) of the LE' protein of the tryptophan operon of E coli, elicited an immune response in steers sufficient to withstand the challenge of exposure to animals with acute FMD. The 58-micrograms dose of viral peptide, composed of a segment of the VP1 from the A12 strain (A12) of FMD virus (FMDV; A12-32dimer) in a tandem repeat configuration of AA137 through 168 and emulsified with oil adjuvant, elicited a serologic response in cattle equivalent to that obtained using conventional whole virus vaccines. Two groups of swine were vaccinated, 1 with the A12-32dimer as used in cattle and 1 with AA131 through 157 from VP1 of the A24 strain (A24) of FMDV (A24-peptide), expressed in the same system as A12-32dimer, but as a single copy per molecule. In swine, the 58-micrograms dose of the A12-32dimer repeated at 28 days was an effective immunogen; all swine were protected against A12 and, in addition, the vaccine protected 50% of the swine against A24. The 29-micrograms dose of A24-peptide, administered according to the same schedule, elicited protection against A24 in 50% of the vaccinates and, in addition, protected 25% of those vaccinates against A12. The serologic response elicited by A12-32dimer against A24 virus was considerably greater than the response elicited by A24-peptide against A12 virus. The evidence of multiple immunogenic epitopes between AA131 and AA168 was evaluated.

Amino Acid Sequence

Use of pulmonary capillary wedge pressure to assess severity of mitral stenosis: is true left atrial pressure needed in this condition?

There is disagreement concerning the use of the pulmonary capillary wedge pressure (in place of left atrial pressure) in assessing the presence and severity of mitral valve disease. This study was done to assess the accuracy and reliability of an oximetrically confirmed pulmonary capillary wedge pressure in measuring the transvalvular pressure gradient and valve area in patients with mitral stenosis. In 10 patients with mitral stenosis (1 man and 9 women; mean age +/- SD 47 +/- 7 years), pulmonary capillary wedge pressure was measured through an 8F Goodale-Lubin catheter with its wedge position confirmed by oximetry (oxygen saturation greater than or equal to 95%). In addition, a transseptal left atrial pressure was measured through a Brockenbrough catheter and left ventricular pressure was measured through a pigtail catheter. The mean and phasic left atrial and pulmonary capillary wedge pressures were similar (mean left atrial pressure 18 +/- 6 mm Hg; mean pulmonary capillary wedge pressure 18 +/- 8 mm Hg; p = NS). When the pulmonary capillary wedge pressure was used without adjustment for time delay, the transvalvular pressure gradient (9.8 +/- 3.3 mm Hg) and valve area (1.5 +/- 0.5 cm2) were significantly different (p less than 0.05) from the values obtained with use of left atrial pressure (7.2 +/- 2.9 mm Hg and 1.7 +/- 0.6 cm2, respectively). In contrast, when the pulmonary capillary wedge pressure was adjusted for the time delay through the pulmonary vasculature, the difference in gradients averaged only 1.7 mm Hg and the mitral valve areas were similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial Function

Office-based system for voice analysis.

There has been recent growing interest in the analysis of various electronically recorded signals as potential tools for objective assessment of vocal dysfunction. In the past, analysis of such signals required an expensive multitrack FM recorder, mainframe computer system, customized software, and significant time commitment. This report describes an adaptation of commercially available components that allow digital recording of multiple electronic signals, storage of data, and subsequent signal analysis using an inexpensive personal microcomputer system. Commercially available software for manipulation and examination signals is discussed as adapted for examination of glottographic and acoustic signals. The relatively inexpensive availability of similar computer systems will, hopefully, encourage assessment of the clinical applications of objective techniques of voice quality.

Analog-Digital Conversion

Identification of virus neutralizing epitopes on naturally occurring variants of type A12 foot-and-mouth disease virus.

Four naturally occurring antigenic variants of foot-and-mouth disease virus type A12 were examined for their capacity to be neutralized by a number of monoclonal antibodies (MAb) which recognize different sites on the virus surface. The VP1 coding region of the RNA genome was sequenced and amino acid changes were determined for the variants. One of the neutralizing sites accounted for the differing antigenic properties of the variants and the epitope was mapped to amino acid residues 150-156 of VP1. Another epitope originally thought to occupy a single site in the area of amino acids 168-179 of VP1 was found on all of the variants. Competitive binding assays did not identify the exact binding site for this monoclonal antibody and the results suggest that the epitope may represent a site more complex than a sequential epitope. A polyclonal antiserum to a 13 kDa fragment of VP1 (residues 55-179) was found to have all the virus reactivity associated with sites located between residues 133-164 of VP1. In contrast, an antiserum to VP1 was found to have additional virus binding sites outside of the 133-164 region of VP1.

Antibodies, Monoclonal

Antigenic comparison of different foot-and-mouth disease virus types using monoclonal antibodies defining multiple neutralizing epitopes on FMDV A5 subtypes.

Thirteen monoclonal antibodies (MAbs) were elicited with A5 Spain-86 virus, the cause of the most recent foot-and-mouth disease virus (FMDV) outbreak in Spain. The MAbs were tested for ability to bind 140S virions and 12S protein subunits by liquid-phase radioimmunoassay (RIA), and to bind VP1 capsid protein by Western immunoblot assay. One of the thirteen MAb was virion (140S) specific, seven recognized 140S and 12S subunits, one bound to 140S, 12S and VP1 and four were 12S specific. These MAbs presented varying binding patterns when tested against different FMDV subtypes and serotypes, indicating the presence of conserved and non-conserved epitopes among FMDV serotypes and subtypes. Neutralization assays, in vivo and in vitro, showed that none of the 140S specific MAbs or 12S specific MAbs were neutralizing, but notably several of the 12S specific MAbs bound to all the different FMDV serotypes and can be useful diagnostic reagent for the detection of FMDV. The remaining MAbs showed varying behavior with the different types tested: not all types to which the MAbs bound were neutralized, demonstrating that the presence of an epitope and subsequent neutralization of infectivity are not necessarily correlated. Five type A12 neutralizing MAbs, previously characterized, have been used in this work. Four bound to A5 Spain-86 virus, but only one neutralized viral infectivity. On the basis of differential reactivity and neutralization among various FMDV subtypes and serotypes, and cross-inhibition binding assays between these MAbs, seven neutralization related epitopes have been defined on A5 Spain-86 virus.

Animals

Analysis of neutralizing antigenic sites on the surface of type A12 foot-and-mouth disease virus.

A series of seven neutralizing monoclonal antibodies (nMAbs) directed against type A12 foot-and-mouth disease virus was used to generate neutralization-resistant variants. Both plaque reduction neutralization and microneutralization assays showed that the variants were no longer neutralized by the nMAbs used to generate them, although some of the variants still reacted with the nMAbs at high antibody concentrations. Results of cross-neutralization studies by both plaque reduction neutralization and microneutralization assays suggested the presence of at least one immunodominant antigenic site on the surface of type A12 foot-and-mouth disease virus, along with evidence of a second antigenic site on the viral surface. Two of the variants had reduced virulence in tissue culture as evidenced by their inability to inhibit cellular protein synthesis and a marked reduction in virus-induced cellular morphological alterations. Nucleotide sequencing of the variant genomes placed three epitopes of the major antigenic site on VP1 and the fourth epitope on VP3 and VP1. The one epitope of the minor site appears to reside only on VP1.

Amino Acid Sequence

The effect of recurrent laryngeal nerve stimulation on phonation in an in vivo canine model.

The present investigation was designed to examine the effect of variation in recurrent laryngeal nerve stimulation (RLNS) on vocal fold vibration. Photoglottography (PGG), electroglottography (EGG), and subglottic pressure (Psub) were measured in seven mongrel dogs using an in vivo canine model of phonation. The PGG, EGG, and Psub signals were examined at three fundamental frequencies (F0) (100 Hz, 130 Hz, and 160 Hz) for RLNS, using a constant rate of air flow. Increasing RLNS, which caused activation of the intrinsic laryngeal muscles, produced a modest increase in F0, a marked increase in Psub, no change in the open quotient (OQ), and an increase in the closing quotient (CQ). Phase quotient (Qp), which describes the interval between opening of the lower and upper fold margins, decreased with increasing RLNS.

Animals