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Biomedical subjects

C A Thomas

Publications and source records attributed to C A Thomas.

At least 91 records · Page 5Linked to original sources

Nuclear RNA transcripts from Drosophila melanogaster ribosomal RNA genes containing introns.

The transcription of ribosomal genes in a cell line (Kc) derived from female D.melanogaster, has been investigated using hybridization probes prepared from restriction fragments of a cloned rDNA repeat with a 5 kb type I [Wellauer et al. (1978) Cell 14, 269-278] intron. Gels, of nuclear RNA that have been transferred to diazotized paper and hybridized with labelled intron sequences, reveal both large (1-10 kb) transcripts and a discrete 325 base species. Berk-Sharp experiments [(1977) Cell 12, 721-732] reveal large transcripts that are homologous to intron sequences and extend into 28S sequences as well. However, while the abundance of 28S transcripts is about 50,000 copies per nucleus [Clark et al. (1977) Genetics 86, 789-800], these long transcripts are only present at 1-2 copies per nucleus and the 325 base species is only 10 times more abundant. In view of the fact that female cells have about 400 rDNA genes, 49% of which have type I introns, one must conclude either that transcription rarely occurs from the genes containing introns (the majority) or these transcripts are processed unusually rapidly. Transcripts homologous to the "non-transcribed spacer" region have been found, but their abundance is no higher.

Animals↗

Characterization of a monoclonal antibody that defines an immunoregulatory T cell subset for immunoglobulin synthesis in humans.

This study characterizes a monoclonal antibody (3A1), and partially characterizes the cell surface antigen and the functional peripheral blood T cell subset that it defines. The 3A1 antigen is present on the surface of several human T cell lines (HSB-2, CEM, MOLT-4, and others) in various amounts but is absent from the T cell line YT4E and all human B cell lines tested. Immunoprecipitation of an HSB-2 extract with 3A1 yielded one specific band with a molecular weight of approximately 40,000 in the presence of reducing agent. With directly fluoresceinated 3A1 antibody, fluorescence-activated cell sorter analysis showed that 85% of peripheral blood E-rosette-positive T cells were positive for the 3A1 antigen. After E-rosette-positive cells had been separated into 3A1+ and 3A1- cell suspensions, the 3A1+ cells helped autologous peripheral blood B cell suspensions toward pokeweed mitogen-driven proliferation and intracytoplasmic Ig production, whereas 3A1-T cells did not. Further, addition of 3A1- cells from some but not all normal subjects to cocultures of 3A1+ cells and B cells actively suppressed intracytoplasmic Ig production. However, the 3A1+T cell subset could be activated by concanavalin A to maximally suppress B cell Ig synthesis in vitro. Thus, the 3A1 antibody defines a major functional subject of peripheral blood T cells and should provide a useful marker for the study of human T cell function.

Antibody Formation↗

Differential binding of fluorescein-labeled lectins to mouse thymocytes: subsets revealed by flow microfluorometry.

Fluorescein-labeled lectins bound to mouse thymocytes were analyzed by flow microfluorometry. This technique has identified several lectins that bind differentially to thymocyte subsets. The most complex fluorescence distributions were obtained using lectins with nominal specificities for galactose or N-acetylglucosamine. Inhibition of binding by sugars confirmed that the fluoresceinated lectins were bound to cells at their carbohydrate binding site. Simultaneous analyses of lectin fluorescence and forward light scatter intensity showed that cell subpopulations of different sizes can exhibit marked differences in the level of binding such that the amount of lectin bound per cell is often independent of cell size. A minor population of dull or unstained cells, delineated by several of these lectins, correlates with the subpopulation of medium-sized thymocytes resistant to in vivo cortisone treatment.

Abrin↗

Polymorphonuclear leukocytes: possible mechanism of accumulation in psoriasis.

Extracts of involved and uninvolved skin from nine patients with untreated psoriasis were studied for chemotactic activity. Psoriatic plaque contains increased amounts of a complement-dependent chemotactic factor that is inhibited by diisopropyl fluorophosphate. This factor may be human skin serine proteinase.

Animals↗

Molecular cloning.

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Base Sequence↗

Molecular cloning of DNA fragments produced by restriction endonucleases Sa1I and BamI.

The highly specific restriction endonucleases Sa1I and BamI produce DNA fragments with complementary, cohesive termini that can be covalently joined by DNA ligase. The Escherichia coli kanamycin resistance factor pML21 has one SalI site, at which DNA can be inserted without interfering with the expression of drug resistance or replication of the plasmid. A more convenient cloning vehicle can be made with the tetracycline resistance factor pSC101, since insertion of DNA either at its single site for Sa1I or at that for BamI inactivates plasmid-specified drug resistance but not replication. To take advantage of this insertional inactivation, pSC101 was joined to a Co1E1-ampicillin resistance plasmid having no Sa1I site, and to a Co1E1-kanamycin resistance plasmid having no BamI site. Chimeras formed with the resulting hybrid vehicles can be identified simply by replica plating. These three vehicles, which all replicate under relaxed control, have been used to clone and amplify Drosophila melanogaster DNA fragments.

Bacillus↗

The actions of restriction endonucleases on lampbrush chromosomes.

Lampbrush chromosomes from oocytes of Notophthalmus viridescens were dispersed in media containing restriction endonucleases isolated from Haemophilus and E. coli. These endonucleases cleave duplex DNAs at specific palindromic sequences of nucleotides, and several sensitive sites occur per micron of DNA. The overwhelming majority of the lateral loops of lampbrush chromosomes are extensively fragmented by these endonucleases, but an occasional pair of loops is refractory. A notable example of loops showing this refractory property are the giant loops on chromosome II in the presence of Hae. These loops, whose DNA-containing axes are several hundred micra long, are sensitive to other nucleases such as EcoB, endonuclease I and pancreatic DNase I; their refractory behavior towards Hae therefore indicates that the sequence sensitive to this particular endonuclease is systematically absent. This anomalous property can be comprehended if it be assumed that the axial DNA of the giant loops consists of tandem repeats of a sequence which happens not to include the sensitive site.

Animals↗