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Biomedical subjects

D S Adams

Publications and source records attributed to D S Adams.

At least 37 records · Page 2Linked to original sources

Performance studies of an enzyme-linked immunosorbent assay for detecting Staphylococcus aureus antibody in bovine milk.

An enzyme-linked immunosorbent assay (ELISA) for detecting Staphylococcus aureus antibody in bovine milk samples was examined for repeatability. A set of 51 bovine milk samples from 4 universities with confirmed culture results was assembled, and a panel of 30 milk samples was randomly selected. When the selected panel was tested at the collection laboratory, there was 97% agreement between the ELISA and the culture test. The panel was tested with the ELISA by the 4 university laboratories. Results were scored by both visual and optical density reader methods. When compared to reference ELISA results, the university laboratory ELISA results showed an agreement of 99.8% for negative samples, 98% for positive samples, and 99% for all samples. Additional studies on 19 milk samples that cultured positive for bacteria other than S. aureus showed 100% specificity. Overall comparison of ELISA and culture results showed high agreement between the 2 techniques. Disagreement appeared to result from explainable differences in antibody and bacterial levels and not from errors in either of the 2 techniques.

Animals↗

The mechanics of notochord elongation, straightening and stiffening in the embryo of Xenopus laevis.

We have examined the biomechanical development of the notochord of Xenopus early tail-bud embryos by: (1) quantifying morphological and mechanical changes in the embryo during stages 20-28, and (2) conducting manipulative experiments to elucidate mechanical roles of various components of the notochord. The notochord, which is composed of a stack of flat cells surrounded by a connective tissue sheath, elongates dramatically and begins straightening between stages 21 and 25. At this time the fiber density in the notochord sheath goes up, the osmotic activity of the notochord cells increases, vacuoles within these cells swell, the internal pressure of the notochord increases 2- to 3-fold, and the flexural stiffness of the notochord rises by an order of magnitude. We suggest that the tendency of the notochord cells to osmotically swell is resisted by the sheath, thereby permitting the internal pressure to rise. This pressure increase results in the greater stiffness that permits the notochord to elongate and straighten without being buckled by the surrounding tissues.

Animals↗

High levels of interferon in synovial fluid of retrovirus-infected goats.

A high level of interferon (IFN) was found in the synovial fluid of goats with naturally acquired caprine arthritis-encephalitis (CAE), a disease caused by caprine arthritis-encephalitis virus (CAEV), a nononcogenic retrovirus of the lentivirus group. Intraarticular injection of CAEV or control inoculum in the joints of affected goats caused increased amounts of IFN production in synovial fluid.

Animals↗

Staphylococcus aureus antigens reactive with milk immunoglobulin G of naturally infected dairy cows.

A 14- to 26-kilodalton fraction of Staphylococcus aureus exoproteins isolated by molecular sieve chromatography and electroelution from polyacrylamide gels was shown to specifically react with antibodies in milk of naturally infected dairy cows. Silver staining of the antigen preparation electrophoresed in polyacrylamide gels showed the strongest reactivity in the 24- to 26-kilodalton region with lesser staining at lower apparent molecular sizes. An enzyme-linked immunosorbent assay was developed to differentiate infected from uninfected cows for diagnostic purposes. Samples from S. aureus-infected cows reacted in the assay, and samples from uninfected cows did not. There was no correlation between numbers of somatic cells in the samples and reactivity to the antigens. Samples from cows infected with coagulase-negative staphylococci did not react with the antigens. It was found, however, that some samples from uninfected cows that were recently postpartum or producing low amounts of milk contained antibodies which bound the antigens. This was believed to be due to transport from blood to the mammary gland of antibodies which were generated by previous intramammary infections or infections at other sites.

Animals↗

Chronic disease in goats orally infected with two isolates of the caprine arthritis-encephalitis lentivirus.

Newborn Saanen goats were orally infected with two distinct biologically cloned isolates of the caprine arthritis-encephalitis lentivirus (CAEV), designated CAEV(63) (Crawford TB, Adams DS, Cheevers WP, Cork LC: Science 207:997, 1980) and CAEV (Co) (Cork LC, Hadlow WJ, Crawford TB, Gorham JR, Piper RC: J Infect Dis 129:134, 1974; Narayan O, Clements JE, Strandberg JD, Cork LC, Griffin DE: J Gen Virol 50:69, 1980). All infected goats seroconverted for antibodies to CAEV, and there were no serologic reversions. Histologic lesions consisted of chronic inflammation of variable incidence and severity in joints and mammary gland. Clinical arthritis, estimated by enlargement of carpi, correlated with severe inflammation of radiocarpal synovium. Statistical evaluations of clinical and histologic parameters indicated that chronic joint disease was significantly more frequent and more severe in the group of goats infected with CAEV(63) than in the group infected with CAEV (Co). Thus, the severity of chronic arthritis is determined in part by the relative pathogenicity of individual CAEV isolates. Analysis of the temporal development of carpal arthritis during the first 3 years of infection indicated that (a) the extent of eventual chronic disease in individual joints was apparent relatively early after infection, (b) joints with severe chronic lesions were clinically enlarged at irregular intervals and (c) the pattern of recurrent arthritis was unique for each joint. Thus, chronic arthritis develops with a progressive course which may be promoted by recrudescent inflammation. Infectious virus was recovered 3 years after infection from selected tissues of 12 of 17 CAEV(63)-infected goats and 11 of 18 CAEV(Co)-infected goats. Virus recovery was independent of the virus isolate used for experimental infection and did not correlate with the severity of lesions.

Administration, Oral↗

A library of trimethylguanosine-capped small RNAs in Physarum polycephalum.

We have constructed a cDNA library for the trimethylguanosine-capped small RNAs (sRNAs) in the acellular slime mold Physarum polycephalum. Capped sRNAs were purified from total cellular RNA of vegetative microplasmodia by preparative immunoprecipitation with anti-trimethylguanosine antibody. The purified RNA was analyzed by polyacrylamide gel electrophoresis. Approx. eleven different capped sRNAs were observed with a size range of 70-204 nucleotides (nt). Based on their approximate sizes, the presence of trimethylguanosine cap, and the presence of a lupus type-Sm antigen, molecules U1-U7 (excluding U3) were identified. Further confirmation of the identity of molecule U1a was established by Northern hybridization, U4a by colony hybridization, and U6 and U7a by direct chemical sequence analysis. Purified capped sRNAs were tailed with oligo(A), and inserted into oligo(dT)-tailed plasmid pCDV1. The cDNAs were used to transform Escherichia coli strain HB101. Approx. 1.9 X 10(5) ampicillin-resistant (ApR) transformants were obtained per microgram of tailed sRNA. Dot-blot hybridization, using Physarum RNA precipitated with anti-cap antibody as a probe, indicated that approx. 94% of the ApR colonies contained recombinant DNAs. The library was screened by colony hybridization using heterologous sRNA probes. Clones hybridizing with heterologous sRNAs U1, U2, U4 and U7 were each represented in the library in approximately the same frequency as their relative abundance in the Physarum sRNA population they were derived from. The insert of one Physarum U4 clone was sequenced and was found to have 57.1% homology with nt 1-91 of the published sequence for rat U4 RNA. A 12-nt 'functional' subdomain of the rat U4 molecule was 83.3% conserved in Physarum U4.

Autoantigens↗

A highly reiterated family of transcribed oligo(A)-terminated, interspersed DNA elements in the genome of Bombyx mori.

A library of low Cot DNA (Cot is the molar concentration of DNA times the incubation time in seconds) from Bombyx mori was used to isolate five independent clones of highly reiterated sequences from the genome of this organism. Sequence analysis revealed that all five clones belong to a single family of repetitive DNA elements, which we have named Bm1, and whose reiteration frequency is approximately 2.3 X 10(4) copies per haploid genome. Probing of a Bombyx genomic library (in lambda phage) with a Bm1 clone reveals that this repetitive sequence is dispersed throughout the genome. The pattern of interspersion was confirmed by Southern blot mapping of a large (270 X 10(3) base-pairs) domain of the chorion locus of Bombyx, where at least 13 independent regions were found to hybridize to Bm1. Four additional Bm1 elements have been sequenced from a 4.8 X 10(3) base-pair genomic fragment containing an early chorion gene. Two of these four elements are bounded by short (4 to 12 base-pairs) direct repeats. The nine Bm1 elements which have been sequenced are greater than 88% homologous to each other, and tend to fall in at least two size classes (253 base-pairs and 450 base-pairs). Seven of the nine Bm1 elements have a short 6 to 10 base-pair oligo(A) sequence at the 3' end. A sequence of about 29 base-pairs at the 3' end, including the oligo(A), shows 86% homology to the equivalent 3'-terminal domain of human Alu family repetitive elements. A 129 base-pair domain at the 5' end of Bm1 shows 66% homology to a Drosophila valine transfer RNA gene; thus the 5' end of Bm1 may contain the split internal RNA polymerase III promoter that is characteristic of most transcribed tRNA-like retroposons. Dot-blot analysis of Bombyx RNA shows that Bm1 DNA is indeed transcribed, and that the transcripts are well-represented in the total RNA of an ovarian-derived permanent cell line and posterior silk glands early in the fifth instar, but are less abundant in the RNA of pupae or silk glands late in the fifth instar.

Animals↗

Acute arthritis in caprine arthritis-encephalitis virus challenge exposure of vaccinated or persistently infected goats.

Goats vaccinated with inactivated caprine arthritis-encephalitis virus (CAEV) developed more severe arthritis after infectious CAEV challenge exposure than did goats vaccinated with tissue culture medium. Arthritis also developed more rapidly in the group vaccinated with inactivated virus. In another experiment, goats with persistent CAEV infection developed acute arthritis after at least 2 injections of infectious CAEV at monthly intervals. In this experiment, the control group consisted of goats with persistent CAEV that were given tissue culture medium. Seemingly, the immune response to CAEV is an important cause of the CAEV-induced arthritis.

Animals↗

The gp135 of caprine arthritis encephalitis virus affords greater sensitivity than the p28 in immunodiffusion serology.

The 135,000 mw glycoprotein (gp135) and the 28,000 mw internal protein (p28) of caprine arthritis encephalitis virus are major viral constituents in precipitin lines formed between crude antigen preparations and sera from infected goats. In testing 307 goat and sheep sera, 118 samples were positive in a gp135 assay and only 82 were positive in a p28 assay. However, some goat sera were found which reacted only with the p28 and therefore testing for antibody against both proteins may be necessary to identify a maximum number of virus infected goats by immunodiffusion.

Animals↗

Isolation and partial characterization of U1-U6 small RNAs from Bombyx mori.

We have used a variety of techniques to characterize the U-series small nuclear RNAs from the posterior silk gland of Bombyx mori. Six molecular species have been identified which correspond to the vertebrate U1-U6 RNAs by the following criteria: (a) presence of the RNAs in ribonucleoprotein particles which can be immunoprecipitated by lupus Sm antisera; (b) presence of a 2,2,7-trimethylguanosine cap, as assayed by immunoprecipitation with anti-2,2,7-trimethylguanosine IgG; (c) size, as assayed by acrylamide/urea gel electrophoresis using HeLa cell U-RNA markers; and (d) primary nucleotide sequence, as determined by chemical/enzymatic cleavage of end-labeled molecules. The high conservation of primary sequence (66-81% homology based on partial sequences) relative to the corresponding vertebrate U-RNAs has permitted unambiguous identification of each molecule. With the exception of two subspecies of U3 RNA, the U-snRNAs of Bombyx exhibit a striking conservation of secondary structure relative to the proposed structures of the U-RNAs of vertebrates. This conservation is best exemplified by several compensatory base alterations that result in the maintenance of hairpin structures. These are particularly evident in U1 and U5 RNAs. Bombyx U3 is interesting in that two subspecies (of a total of four that were sequenced) diverge considerably in sequence (and presumably in structure) relative to the U3 RNA of vertebrates. The most abundant U-RNAs in the posterior silk gland appear to be U1 and U2, while U3-U6 are present in relatively small amounts.

Animals↗

Identification of caprine arthritis-encephalitis retrovirus proteins in immunodiffusion precipitin lines.

Precipitin lines formed between serum from a goat infected with caprine arthritis-encephalitis virus (CAEV) and radiolabelled viral proteins in polyethylene glycol-concentrated culture medium were excised from immunodiffusion (ID) plates and analysed by polyacrylamide gel electrophoresis. The two major precipitin lines contained the 135 000 mol. wt. glycoprotein (gp 135) and the internal 28 000 mol. wt. structural protein (p28). This method obviates the use of purified proteins or monospecific antisera to positively determine viral constituents in ID precipitin lines formed between a crude antigen preparation and antiserum against whole virus.

Animals↗

Antigenic cross-reactivity between caprine arthritis-encephalitis, visna and progressive pneumonia viruses involves all virion-associated proteins and glycoproteins.

Antigenic relatedness between the virion-associated proteins of caprine arthritis-encephalitis, visna and progressive pneumonia viruses was examined. Antigenic cross-reactivity was assessed by immunoprecipitation of disrupted, radiolabelled virus with goat, sheep and rabbit antisera, followed by resolution of the immunoprecipitation products by SDS-polyacrylamide gel electrophoresis. The results indicate that antigenic cross-reactivity between the caprine and ovine virus isolates involves all of the major virion-associated proteins and glycoproteins. The common antigenic determinants exhibited by virion structural proteins are immunogenic in goats, sheep and rabbits.

Animals↗

Preliminary characterization of a serum viral inhibitor in goats.

A serum viral inhibitor (SVI) was isolated from goats and partially characterized. The inhibitor prevented the cytopathic effects of vesicular stomatitis virus, encephalomyocarditis virus, and a caprine herpesvirus, indicating broad antiviral activity. The SVI was distinct from interferon because SVI did not induce an antiviral state in cells (ie, lack of protection of SVI-treated cells from virus challenge). The SVI had activity on heterologous cells, including human, bovine, and ovine cells. The lack of antiviral activity in mouse cells indicated that SVI was not an antibody. Like fibroblast interferon, however, SVI was sensitive to trypsin, was acid stable at pH 2 and 4 C for 4 days, was heat stable at 56 C for 1 hour, and could not be sedimented by centrifugation at 100,000 X g for 4 hours.

Animals↗

Global survey of serological evidence of caprine arthritis-encephalitis virus infection.

Using caprine arthritis-encephalitis virus antigen in the agar gel immunodiffusion test, 3729 serum samples from goats in over 112 locations around the world were tested for precipitating antibodies. Over 90 per cent of the 1265 positive samples came from Canada, France, Norway, Switzerland and the USA, all of which had 65 per cent reactors or greater. Fiji, Great Britain, Kenya, Mexico, New Zealand and Peru had fewer than 10 per cent positive samples; the majority of these could be traced to importations of goats from countries where there was a high occurrence of precipitating antibody. Somalia, Sudan and South Africa had no reactors among 306 samples. No reactors were found among 1116 samples from domestic and indigenous goats which were known to have had no contact with imported goats from countries which had a high occurrence.

Africa↗