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Biomedical subjects

J R Duncan

Publications and source records attributed to J R Duncan.

At least 91 records · Page 5Linked to original sources

Determinants of off-road vehicle transmission 'shift quality'.

This study was conducted to determine the relationship between factors associated with a tractor transmission shift and the tractor operator's judgement of the quality of the shift. The specific objective was to measure experienced tractor operators' ratings of shift 'acceptability' as peak acceleration, peak jerk and direction of initial acceleration were varied at two levels of background vibration. A six-degree-of-freedom vehicle motion simulator in the Deere & Company Technical Center's human factors laboratory was used to simulate the background vibration and to control the experimental variables. The key results are: (1) The most important factor influencing 'shift quality' is the peak acceleration experienced during the shift. (2) Background vehicle vibration experienced by the tractor operator affects his perception of 'shift quality'. Higher background vibration masks the acceleration experienced during a shift and makes the shift more acceptable than when experienced at a low level of background vibration. (3) Peak jerk experienced during a shift has a negligible effect on 'shift quality'. (4) Upshifts and downshifts of the same peak acceleration, peak jerk, and background vibration receive the same general rating of 'shift quality'.

Journal Article↗

Protein constituency of unbound fraction of bovine serum applied to anion exchange columns at different pHs and molarities with or without ethylenediamine tetraacetic acid. Application to preparation of bovine IgG1.

Ethylenediamine tetraacetic acid disodium salt (EDTA) was found to be an important addition to anion exchange buffers in terms of the profile and amounts of eluted bovine serum proteins. Hydrogen ion concentration and buffer composition were important when different types of anion exchanger (DEAE 52 cellulose and QAE Sephadex A 50) were used for separation of bovine serum proteins as functional groups of the different anion exchangers did not behave similarly and were therefore not interchangeable. These findings applied to the purification of bovine serum IgG1 amounting to 15 to 25% of the total IgG1 was accomplished using QAE Sephadex A 50 anion exchange chromatography. This was followed by absorption of the IgG1 fraction with Staphylococcus aureus containing protein A to remove minor IgG2 contaminants and gel filtration to exclude traces of the third component of complement (C3).

Animals↗

Preparation of heavy chain specific antisera to bovine IgA, IgM, IgG1 and IgG2.

Goats, guinea pigs and rabbits were immunized with bovine IgM or with intact molecules, heavy chains, Fc portions or light chains of bovine IgG1 and IgG2. Rabbits and guinea pigs were immunized with bovine secretory IgA. Goats and guinea pigs produced heavy chain specific antisera to intact IgM whereas rabbits produced anti-light chain antibody and in one instance anti-alpha 2-macroglobulin antibody in addition to the anti-mu response. Goats and guinea pigs produced antisera to bovine IgG1 and IgG2 and their Fc portions that needed little absorption to render them monospecific for the heavy chain. In addition to antibody to the heavy chains, rabbits produced anti-light chain antibody when immunized with intact IgG1 or IgG2 molecules. These latter sera, and those produced by rabbits immunized with Fc portions of IgG1 or IgG2 required extensive absorption before they were monospecific for their respective heavy chains. Heavy chains were poor immunogens in all three species. Rabbits immunized with IgA produced both anti-alpha and anti-light chain antibodies while guinea pigs produced sera with antibody activity to the alpha chain only.

Animals↗

Secretion and degradation of mutant leucine-specific binding protein molecules containing C-terminal deletions.

The leucine-specific binding protein (LS-BP), a periplasmic component of the Escherichia coli high-affinity leucine transport system, is initially synthesized in a precursor form with a 23 amino acid N-terminal leader sequence that is removed during secretion of the protein into the periplasm. Using in vitro mutagenesis, deletion mutants of the LS-BP gene have been constructed with altered or missing amino acid sequences in the C-terminal portion of the protein. These altered binding proteins exhibited normal processing and secretion but were rapidly degraded in the periplasmic space. In the presence of an uncoupler of the transmembrane potential (CCCP) the precursor forms accumulated in the membrane and were protected from degradation. The altered binding proteins also were secreted by spheroplasts of E coli, after which they were easily detected.

Bacterial Proteins↗

Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3.

Antigenic phenol-phase soluble lipopolysaccharide isolated from Brucella abortus 1119-3 by hot phenol-water extraction was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, controlled hydrolysis, periodate oxidation, methylation, and 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as an unbranched linear homopolymer of 1,2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl residues. The serological reactivity of bovine antiserum to B. abortus 1119-3 with the lipopolysaccharides of Yersinia enterocolitica serotype O:9 and Vibrio cholerae species has now been related to the occurrence of 1,2-linked N-acylated 4-amino-4,6-dideoxy-alpha-D-mannopyranosyl units in the O-chain polysaccharides of their lipopolysaccharides.

Antigens, Bacterial↗

Serological confirmation of Brucella abortus and Yersinia enterocolitica O:9 O-antigens by monoclonal antibodies.

Murine monoclonal antibodies that bind the O-antigens of Yersinia enterocolitica serotype O:9 and Brucella abortus 1119-3 were generated after immunization of BALB/c mice with killed, whole cells. Highly purified lipopolysaccharide preparations from each organism were used to screen for antigen-specific antibodies. Immunization with B. abortus cells induced 56 antigen-specific hybrids, and 10 of the highest antibody-producing clones were selected for further study. Seven of these clones secreted immunoglobulin G, and three secreted immunoglobulin M antibodies. Immunization with Y. enterocolitica cells resulted after fusion in 76 antigen-specific hybrid cell lines; from these, seven immunoglobulin G-secreting clones were selected for study. The serological cross-reactivity of the B. abortus and Y. enterocolitica O-antigens was established by enzyme-linked immunosorbent assay, immunoprecipitation, and agglutination tests with the monoclonal antibodies induced by each bacterium. This serological cross-reactivity is consistent with the structural identity of the two O-antigens established by chemical analysis.

Antibodies, Monoclonal↗

A computer-interfaced photometer and systematic spacing of duplicates to control within-plate enzyme-immunoassay variation.

A Multiskan photometer for reading microtiter plate enzyme immunoassays was linked with a time sharing computer to facilitate control of assay variation and analysis of results. The interface that converted photometer output to RS-232-C format required changes to divide the output into segments short enough for input to the computer. To measure within-plate variation and investigate how the method of allocating sample duplicates to plate wells may affect the estimation of sample variance, uniformity tests were conducted with 47 plates. Coefficients of variation (CV) among wells within-plates ranged from 4.6 to 20.7% and in two-thirds of the plates exceeded 10%. Duplicates allocated to adjacent wells (method 1) gave consistently higher CV for sample means than duplicates allocated to opposite plate quadrants (method 2). In general, the CV by method 2 was about 30% smaller than that by method 1. Analysis of variance confirmed the effectiveness of the quadrant pattern of duplicate allocation as a method of controlling variation that arises from well position effects.

Analysis of Variance↗

Aplastic crisis due to parvovirus infection in pyruvate kinase deficiency.

A thirteen-year-old boy with congenital haemolytic anaemia due to pyruvate kinase (PK) deficiency had an aplastic crisis. A serum parvovirus-like virus (SPLV) was demonstrated in the blood by electron microscopy and, subsequently, IgM and IgG antibodies to the prototype SPLV B19 were detected. In an attempt to define the level of erythropoiesis that is involved in parvovirus-induced bone marrow suppression, the levels of circulating early erythroid progenitors (burst forming units erythroid, BFU-E) were monitored during the crisis and recovery period. The virus-containing plasma inhibited the formation of BFU-Es from non-immune subjects and this effect was neutralised by convalescent serum. Colony forming units granulocyte-macrophage (CFU-GM) were also inhibited but this was probably non-specific since neutralisation did not reverse the effect. These experiments, together with the clinical data, suggest a selective effect of SPLV at the stage of erythroid progenitors.

Adolescent↗

Marek's disease virus-induced transient paralysis: a comparison of lesions in susceptible and resistant lines of chickens.

A study was conducted to determine the causative lesion of Marek's disease virus-induced transient paralysis. Brain histologic lesions were quantitated and compared in genetically resistant and susceptible inbred chickens. Comparisons were made 11 days post inoculation (PI) of virus when clinical signs were evident in susceptible birds and on day 17 PI when these signs had remitted. Clinically-affected birds from the susceptible line on day 11 PI had consistent evidence of neuropil vacuolation compatible with brain edema; two of these birds also had vasculitis. These lesions were absent in both clinically-recovered birds from this line evaluated on day 17 PI and genetically resistant birds studied on day 11 PI. Our findings suggest that the causative lesion of transient paralysis is vasogenic brain edema.

Animals↗

Comparison of pyrimidine 5'nucleotidase activity in erythrocytes of sheep, dogs, cats, horses, calves, and Mongolian gerbils.

Pyrimidine 5'nucleotidase (P5N) activities of erythrocytes for Mongolian gerbils, cats, dogs, sheep, horses, and calves were measured, using a radiometric technique with [14C]cytidine monophosphate as the substrate. Erythrocytes of gerbils had the highest activity [1,177.1 +/- 133.6 mU/g of hemoglobin (Hb)]. Feline erythrocytes had 327.4 +/- 204.4 mU/g of Hb. Canine erythrocytes had 148.0 +/- 19.8 mU/g of Hb. Ovine erythrocytes (44.3 +/- 20.9 mU/g of Hb), equine erythrocytes (30.0 +/- 15.9 mU/g of Hb), and bovine erythrocytes (14.1 +/- 6.9) had relatively low P5N activity. The P5N activity was approximately proportional to the reticulocyte percentage of and inversely proportional to the mean erythrocyte life span in these species.

5'-Nucleotidase↗

Possible site of zinc control of hepatoma cell division in Wistar rats.

The effect of zinc on the growth of a transplantable DAB hepatoma in young male Wistar rats was determined. Both a zinc deficiency (less than 0.5 microgram/g feed) as well as high levels of dietary zinc (500 micrograms/g feed) significantly reduced tumor growth. Both high- and low-zinc diets resulted in reduced activity of the salvage pathway of thymidine synthesis as well as reduced 32PO4 incorporation into DNA and diminished DNA polymerase activity. Blockage of the de novo pathway of DNA synthesis by the folate antagonist methotrexate (MTX) resulted in greatly increased flux through the thymidine salvage pathway and increased DNA polymerase activity but decreased 32PO4 incorporation in the transplantable hepatomas in Wistar rats fed normal zinc diets (50 micrograms/g feed). MTX had the effect of reducing all these activities in the groups fed low- and high-zinc diets. These data suggested a site of action of zinc associated with the salvage pathway of thymidine synthesis.

Animals↗

Neutrophil movement in selected canine skin diseases.

Neutrophilic movement was studied in dogs with atopy, 3 bacterial pyoderma syndromes, flea allergy dermatitis, and generalized demodicosis. Random neutrophil movement was decreased in generalized demodicosis, and this deficit correlated with the extent of body surface skin lesions. Neutrophilic chemotactic movement was decreased in 2 bacterial pyoderma syndromes: bacterial pyoderma, and staphyloccocal pyoderma. Although serum samples from diseased dogs showed no significant differences in chemoattractive qualities from normal pooled canine serum, control canine neutrophils incubated in demodectic dog serum showed decreased chemotactic responsiveness. It was postulated that demodectic dog serum had an inhibitor or deactivator of neutrophil chemotaxis.

Animals↗

Pyrimidine-specific 5' nucleotidase activity in bovine erythrocytes: effect of phlebotomy and lead poisoning.

Erythrocyte pyrimidine-specific 5' nucleotidase (PY5'N) (E.C. 3.1.3.5) was measured in healthy, anemic, and lead-poisoned calves to determine whether low activity of PY5'N is associated with the propensity of cattle to develop basophilic stippling of erythrocytes. Low activity of PY5'N has been associated with basophilic stippling of erythrocytes in persons with inherited hemolytic anemia and with lead poisoning. A radiometric technique, using [14C]cytidine monophosphate as the substrate, was used to measured PY5'N activity. The erythrocytes from 4 healthy calves had much lower activity (mean of 7.1 +/- 1.6 nmols of [14C]cytidine monophosphate hydrolyzed/min/g of hemoglobin) than has been reported for human erythrocytes. The pH response curve of bovine PY5'N was similar to that of the human enzyme, with maximal activity around pH 7. Experimental hemorrhagic anemia in these calves increased PY5'N activity 6-to 7-fold, with peak activity occurring concomitantly with maximum reticulocytosis. Two of the calves were then given lead per os, and the PY5'N activity decreased within 24 hours to base-line values. In the 2 other calves not given lead, the PY5'N activity declined slowly, but did not reach base-line values after 14 days.

5'-Nucleotidase↗

Demonstration that nonspecific bovine Brucella abortus agglutinin is EDTA-labile and not calcium-dependent.

The binding site of B. abortus involved in nonspecific or false-positive agglutination by bovine serum was shown to be Ca++-independent in that binding of Ca++ by the chelating reagent DP2TA did not decrease titers. The reduction in agglutination by EDTA, EGTA, and DPTA was thought to be related to a common structural characteristic of these chelating reagents not exhibited by EDDA and DP2TA or to a lesser extent by DTPA. Removal of antigenic entities from the cell by the chelator was excluded as the cause of agglutination inhibition because agglutination by nonspecific IgM was not influenced by repeated washing of B. abortus by EDTA. The action of the chelators on the cell-agglutinin interaction was hypothesized to be a result of their competition with a receptor site on the cell for binding of the nonspecific or biochemically altered bovine IgM.

Agglutinins↗

Effect of sample preparation on basophilic stippling in bovine blood smears.

The effect of sample preparation on the amount of basophilic stippling of erythrocytes (BSE) was studied using blood from a calf with chronic experimental lead poisoning. The combination of EDTA anticoagulation and rapid drying of the blood smear resulted in the most BSE. Alcohol prefixation reduced BSE. Wright-Leishman stain was better than Wright stain in demonstrating BSE.

Journal Article↗