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Biomedical subjects

B J Thompson

Publications and source records attributed to B J Thompson.

At least 19 recordsLinked to original sources
PubMed1985

Biosynthesis of fatty acids by lactating human breast epithelial cells: an evaluation of the contribution to the overall composition of human milk fat.

The objective of this study was to characterize the fatty acid biosynthetic pathway of the lactating human breast. Mixed cell populations, obtained by centrifugation of human milk, were enriched in breast epithelial cells by a selective adsorption procedure. Confirmation of the identity of the breast epithelial cells was obtained immunohistochemically. These viable breast epithelial cells incorporated radioactively labeled acetate predominantly into fatty acids with less than 16C atoms. The presence of the two key enzymes characteristic of the medium-chain fatty acid biosynthetic pathway of nonruminants, fatty acid synthetase, and thioesterase II, was demonstrated both qualitatively, by immunohistochemistry, and quantitatively, by enzyme assay. The results indicate that the lipogenic system of the human breast is qualitatively very similar to that of rats, mice, and rabbits, which also secrete milk fats containing medium-chain fatty acids. Quantitatively, however, the mammary fatty acid biosynthetic pathway appears to be less active in humans than in these other species.

Adult↗
PubMed1984

Thioesterase II, a new marker enzyme for human cells of breast epithelial origin.

Metabolic, enzymologic, and immunohistochemical techniques have been used to show that a human cell line of breast epithelial origin synthesized medium chain fatty acids via the ubiquitous fatty acid synthetase and a mammary-specific chain-terminating enzyme, thioesterase II. Previous studies in our laboratory with rodents indicated that thioesterase II is expressed exclusively in mammary epithelial cells, an observation consistent with the physiologic role of the enzyme in milk fat synthesis. Results of the present study suggest that the enzyme exhibits a similar cell specificity in its expression in humans and that the specificity is maintained in normal and neoplastic tissues. Thus thioesterase II was detected immunohistochemically in normal human breast epithelia derived from both lactating and nonlactating breast tissue, in cultured cells derived from both primary breast epithelial tumors and from a metastatic tumor of breast origin, and in several human breast epithelial cell lines; the enzyme could not be detected in HeLa cells, in a colon carcinoma, or in a mammary myoepithelial cell line. These findings raise the possibility that thioesterase II may be of use as a diagnostic tool to identify human tumors of breast epithelial origin.

Adolescent↗
PubMed1982

Lack of coordinated regulation of lipogenic enzymes in a human breast cell line, SKBr3.

A human breast cell line has been identified which contains prodigious levels of fatty acid synthetase but has a very low capacity for lipogenesis from glucose, lactate or acetate. The fatty acid synthetase from this cell line appears to be structurally and functionally normal, and the low lipogenic capacity of the cells appears to be due to the low activities of other lipogenic enzymes, notably acetyl-CoA carboxylase. Thus, the SKBr3 cell line appears to lack the long-term coordinated control of acetyl-CoA carboxylase and fatty acid synthetase commonly observed in normal lipogenic tissues.

Acetates↗
PubMed1982

Localization of thioesterase II, the chain-length regulatory enzyme of milk fatty acid synthesis, in rat mammary gland epithelial cells.

Two approaches were used to establish the intercellular distribution of fatty acid synthetase and thioesterase II in the lactating rat mammary gland. Thioesterase II is the chain-length regulatory enzyme in the biosynthesis of the medium-chain fatty acids characteristic of milk fat. Using immunohistochemical techniques, immunoreactive fatty acid synthetase was found in both mammary adipocytes and epithelial cells; in contrast, immunoreactive epithelial cells were isolated from lactating rat mammary glands after digestion with collagenase and thermolysin, and their lipogenic activity was studied using isotopically labelled acetate. Consistent with the immunohistochemical data, adipocytes synthesized exclusively long-chain fatty acids whereas epithelial cells synthesized predominantly chain fatty acids. The results indicate that the capacity for synthesis of medium-chain fatty acids is a unique property of the epithelial cell component of the mammary gland.

Animals↗
PubMed1981

Purification and properties of fatty acid synthetase from a human breast cell line.

A human mammary epithelial cell line (SKBr3) has been identified in which fatty acid synthetase constitutes up to 28%, by weight of the cytosolic proteins. The enzymes has been purified to near homogeneity from this cell line and some of its properties studied. In common with fatty acid synthetases from other animal tissues, the enzyme is a 480 000 dalton dimer of similar molecular weight subunits, it synthesizes predominantly palmitic acid and is inactive in the absence of free coenzyme A. The kinetic properties and amino acid composition of the enzyme are also similar to those of fatty acid synthetases from various tissues of other animals. Appreciable structural resemblance between human and rodent fatty acid synthetases is indicated by studies on the immunological cross-reactivities of these enzymes.

Amino Acids↗
PubMed1979

Isolation of F-actin filaments. Comparison of F-actin filament preparations from normal and dystrophic mouse muscle.

To investigate the dystrophic influence on the characteristics of actin, a method for the isolation of F-actin filaments from the skeletal muscle of small sizes, i.e., less than 0.5 g, was devised. In this method, minced muscle was treated with collagenase and hyaluronidase, and the isolated filaments were washed with adenosine triphosphate (ATP). Upon examination in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the ATP-washed filaments showed a protein component identical in mobility to actin in untreated myofibrils or to that prepared by the conventional method. Electron microscopic appearances of the filaments were similar to those of F-actin filaments described in the literature. The dimensions of the filaments were 0.5--2.5 micrometer in length and 60--70 A in diameter. The ability to activate the Mg-adenosine triphosphatase or myosin was found to be Ca2+ independent. In all aspects of the above characteristics, the filaments from leg muscles of 129/Re dydy dystrophic mice and their litter mates were observed to be identical.

Actins↗
PubMed1978

Keratosis follicularis spinulosa decalvans. An infant with failure to thrive, deafness, and recurrent infections.

A 10-month-old male infant had keratosis follicularis spinulosa decalvans, an X-linked dominant disorder. His cutaneous abnormalities consisted of generalized hyperkeratosis, spiny follicular papular lesions, universal alopecia, and hypoplastic nails. Ocular changes characteristic of the disease were also present. Unusual findings included deafness, failure to thrive, predisposition to bacterial infections without demonstrable immune defect, and transient hepatomegaly with abnormal liver function studies.

Adult↗
PubMed1976

Kinetics of branch migration in double-stranded DNA.

The rate of branch migration in double stranded DNA has been measured by the use of a unique substrate formed by the action of the EcoRI restriction endonuclease on the dimeric figure-8 configuration of the replicative form DNA of phage G4. The figure-8 and the X-form derived from it contain a junction of the kind postulated to occur in the Holliday structure and to be an essential feature of a number of models of recombination. In the X-form this junction can branch migrate to an irreversible terminal configuration consisting of two linear monomers. The disappearance of X-forms was measured by electron microscopy. A treatment of branch migration as a random walk process was developed to permit the determination of the rate of the intrinsic process, a step movement of the junction by a distance of one base pair. A value of about 6 kilobase pairs per sec at 37 degrees was obtained.

Bacteriophages↗