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Biomedical subjects

W A Thomas

Publications and source records attributed to W A Thomas.

At least 73 records · Page 4Linked to original sources

Two chick embryonic adhesion systems: molecular vs tissue specificity.

We have investigated the adhesive properties of cells from several neural and nonneural chick embryonic tissues dissociated using modifications of the standard dissociation procedures employed routinely in this laboratory to obtain retinal cells. Each of these tissues (7-day optic tectum, retina, and heart, and 3.75-day limb bud) displayed both Ca++-dependent (CD) and Ca++-independent (CI) aggregation, the relative rates of which differed from tissue to tissue. In every case, cells prepared so as to display one mode of aggregation or the other cross-adhered readily to cells--regardless of tissue origin--displaying the same mode of aggregation. Cross adhesion was negligible between cells--even from the same tissue--prepared so as to display different modes of aggregation. Anti-retinal Fab molecules which inhibit selectively either the CI or CD aggregation of retina cells strongly inhibited the corresponding aggregation of optic tectum cells, but had no effect upon the aggregation (CI or CD) of heart cells. These results demonstrate the existence in the tissues examined of dual adhesion mechanisms similar in Ca++ dependence and recognition properties to those of the retina, but showing certain immunological distinctions from the latter. The immunological relationship among the adhesion mechanisms from the various tissues is under continuing investigation.

Animals↗

Circadian variation of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in swine liver and ileum.

The temporal variation of HMG-CoA reductase activity in the liver and intestine of swine was investigated. The thin-layer chromatographic method widely used in the assay of the reductase was successfully applied to the porcine enzymes. Parallel circadian rhythms were demonstrated in both hepatic and ileal reductases from mash-fed animals. Peak activity occurred approximately 6 hr after feeding, 2.7-fold over the basal level in the liver, and 1.6-fold in the ileum. A milk-cholesterol diet caused a marked depression of both rhythms (90% in liver, 50% in ileum); however, the hourly variation in activity persisted in both organs. Cholestyramine was found to elevate hepatic activity (2.7-fold throughout the rhythm) without affecting that of the intestine. Clofibrate had no effect on either enzyme at any time during the cycle despite a 34% reduction in serum cholesterol concentrations.

Animals↗

A twelve-channel automatic device for continuous recording of cell aggregation by measurement of small-angle light-scattering.

We describe here a 12-channel aggregometer, evolved from the instrument described by Beug & Gerisch in 1972, which records the course of aggregation or agglutination of cells or other particles by following the changes in light-scattering of the aggregating suspension. The instrument incorporates a simple memory system for condensing the data, introduces an improved cuvette design, and can be adjusted in its responsiveness to light-scattering by particles of different sizes. In our aggregometer, a vertical wheel in an opaque thermostatted chamber accommodates up to 12 cuvettes each containing a cell suspension and an air bubble. The constant rising of the bubble as the wheel rotates causes the stirring action which promotes aggregation. At a certain point in its rotational path, each cuvette is penetrated by a beam of light focused upon an absorbing beam stop centred in front of a photomultiplier tube. Particles suspended in the cuvette scatter a portion of the light beyond the beam stop into the photomultiplier tube to produce an electrical pulse proportional to the photon input. Collection of these pulses with the wheel in continuous motion avoids any disturbance of the course of aggregation. The pulses are routed to memory circuits for summation and eventual recording by a 12-channel printing potentiometer which automatically colour-codes and numbers each curve. The cuvettes consist of 2 glass microscope coverslips affixed with silicone grease over a hole in a thin, stainless steel blank. They are very durable and are easily dismantled and reassembled for cleaning. The coverslips are replaced after each use. The discoid chamber of our cuvette permits uniform circulation of the bubble, which in turn causes gentle stirring of the aggregating suspension at a rate that is a direct and continuous function of rev/min. Measurement of light-scattering at small angles (3--5.5 degrees) provides great sensitivity to the disappearance of single cells and progressively less sensitivity to the coalescence of aggregates of progressively larger size. The signal generated by the photomultipler tube decreases as aggregation proceeds. At 32 rev/min, one data point per channel, representing the sum (average) of 24 successive measurements, is printed every 45 s. In the course of a 60-min assay under standard conditions, up to 23040 individual readings can be automatically taken, summed and plotted. In a comparison of 8 identical samples run simultaneously, the printed values after 60 min agreed with a standard deviation of +/- 2%.

Animals↗