Avian mycoplasma identification using polymerase chain reaction amplicon and restriction fragment length polymorphism analysis.

A general mycoplasma polymerase chain reaction (PCR) was used to generate amplicon (DNA amplification product) from nine avian mycoplasma species. The PCR amplicons were reacted with 24 restriction enzymes, and the electrophoretic patterns of restriction fragment length polymorphism (RFLP) were evaluated for differences between species of mycoplasms. Four (DraI, MseI, RsaI, Tsp5091) of the 24 restriction enzymes cut the PCR amplicon of all nine mycoplasma species. The nine avian mycoplasma species could be distinctly differentiated using the RFLP analysis of the PCR amplicon.

Investigation of the nature and specificity of antinuclear antibody in dogs.

A liquid phase radioimmunoassay was used to test sera from 34 dogs in 3 categories, determined by presence of antinuclear antibody (ANA) and disease, for antibodies against native canine DNA and dAdT, a synthetic double-stranded DNA analog. Antibodies to dAdT were absent in healthy dogs which, as a group, had levels of DNA antibodies consistent with those reported for healthy persons. Canine patients with a variety of illnesses, but which remained ANA negative, had slightly increased levels of binding of DNA and dAdT. As a group, ANA-positive dogs had significantly increased binding of dAdT and native DNA, which was shown to be mainly, but not entirely, double stranded. In the ANA-positive group, no correlation was found among ANA titer, % DNA binding, and % dAdT binding, indicating that these 3 procedures detect antibodies with differing specificities. In dogs, ANA are heterogeneous in antigenic specificity. Antibodies to double-stranded nucleic acid in dogs do not appear to be as specific for systemic lupus erythematosus as they are reported to be in persons with autoimmune disease.

Comparative aspects of nuclear proteins and nuclear composition of cultivated embryonic, neonatal and neoplastic rat brain cells of glial origin.

Differences in nuclear composition and variations in the types of acid soluble nuclear proteins were identified when cultivated neoplastic and nontransformed rat brain cells were compared. HeLa S-3 cells were used as a biological reference in these studies. The nuclei of anaplastic glioma cells were found to contain more total nuclear protein and greater amounts of nuclear RNA, than the nuclei of nontransformed embryonic and neonatal rat brain cells. Densitometer profiles of samples developed by polyacrylamide gel electrophoresis indicated that all three major histone classes were present in the nuclei of the four different cell types examined. Each type of cell was grown in the presence of 3H-lysine or 3H-lysine plus 14C-arginine to further characterize both the histones and the acid soluble nonhistone proteins. Neoplastic and nontransformed cells contained similar quantities of histones H4, H3, H2A and H2B. In contrast, transformed cells were found to contain two dominant subspecies of lysine rich H1 histone, while only one histone H1 subcomponent was extracted from the nuclei of the embryonic and neonatal rat brain cells. Nuclei isolated from HeLa cells and anaplastic glioma cells also contained increased varieties and larger quantities of acid soluble nonhistone nuclear proteins.