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Biomedical subjects

D F Waugh

Publications and source records attributed to D F Waugh.

At least 19 recordsLinked to original sources
PubMed1985

Effects of calcium binding and of EDTA and CaEDTA on the clotting of bovine fibrinogen by thrombin.

Studies were carried out at pH 7.0 and gamma/2 0.15 before addition of CaCl2 or EDTA. Clotting time, tau, at 3.03 microM fibrinogen and 0.91 u/ml thrombin was determined for equilibrium systems. With added Ca2+, tau decreases, from tau 0 at 0 added Ca2+ (mean, 29.7 +/- 3 s), by approximately 3 s at 5 mM added Ca2+. With added EDTA, tau increases sigmoidally from tau 0 at 0 EDTA to a maximum (mean tau m = 142 +/- 23 s) at approximately 200 microM EDTA. tau then decreases slightly to a minimum at approximately 1.3 mM and finally increases to infinity at approximately 10 mM EDTA. Between 0 and 1.3 mM EDTA, effects on clotting time are completely reversed by adding Ca2+ and, after equilibration at 400 microM EDTA, tau is independent of EDTA concentration. Thus, up to 400 microM EDTA, effects on clotting time are attributed to decreasing fibrinogen bound Ca2+. Between 5 mM Ca2+ and 200 microM EDTA it is assumed that an equilibrium distribution of fibrinogen species having 3, 2, 1, or 0 bound calcium ions is established and that a clotting time is determined by the sum of products of species fractional abundance and pure species clotting time. Analysis indicates that pure species clotting times increase proportionately with decreasing Ca2+ binding, binding sites are nearly independent, and the microscopic association constant for the first bound Ca2+ is approximately 4.9 X 10(6) M-1. Effects of adding Ca2+ at times t1 after thrombin addition to systems initially equilibrated at 200 microM EDTA were determined. Analysis of the relation between tau and t1 indicates that as Ca2+ binding decreases, rate constants for release of B peptides decrease less than those for release of A peptides. As EDTA concentration is increased above 1.3 mM, inhibitory effects of EDTA and CaEDTA progressively increase.

Animals↗
PubMed1978

Interactions of bovine thrombin and plasma albumin with low-energy surfaces.

Surface configurations are vessels fabricated from tubing and plate, films deposited on the surface of vessels, and beads confined in vessels. The average association constant between thrombin and sites on commercial poly(methyl methacrylate) surface (Lucite) is near 4 X 10(8) liters/mole at 22 degrees C, pH 7.0, and ionic strength 0.15. Depending on Lucite composition, average adsorption U, in molecules/cm2 of apparent solution-surface interface, ranges from 0.7 to 8.8 X 10(11). Analysis based on the assumptions that solution dimensions are preserved, adsorption is random, and surface rearrangement is negligible indicates a paucity of surface sites. Plasma albumin competes with thrombin for surface sites. Attempts to detect, by thrombin adsorption, the presence of free sites at 4.5 X 10-9M albumin or the displacement of bound albumin indicate an albumin-site association contrast greater than 1.6 X 10(9). Cross-linked poly(methyl acrylate) bead surface has U less than 5 X 10(10). In contrast to acrylic resins are silicone gum, polypropylene, and polyisobutylene, for which U ranges from 15 to 20 X 10(11). Analysis as above indicates that sites are of frequent occurrence. Material composition suggests that thrombin can interact with nonpolar groups. Further characteristics of low-energy surfaces are that progressive surface denaturation is small and there is a large variance between nominally equivalent configurations.

Absorption↗
PubMed1976

Effects of matrix contact during gel filtration of human platelets in plasma.

When gel filtration is used to transfer platelets from plasma into an established environment, alterations in platelet characteristics may result from the change in environment or from the effects of platelet contact with the gel matrix. To approach the problem of evaluating the relative contributions from these sources, a Sepharose 2B matrix was employed and platelets transferred from citrate anticoagulated PRP into autologous PPP to yield plasma-GFP. Platelet recoveries averaged 93%. PRP: plasma-GFP pairs were found to be indistinguishable with respect to: morphology; ADP, thrombin or collagen-induced aggregation response; uptake of 5-hydroxytryptamine (5-HT) or adenosine; and thrombin or collagen-induced release of accumulated 5-HT or adenosine. Pairs are distinguishable by prostaglandin E2 synthesis assayed immediately after filtration.

Adenosine↗
PubMed1976

Effects of ions on ADP-induced aggregation of bovine or human platelets.

Effects of divalent cations on ADP-induced aggregation response were examined. Bovine platelets were transferred by Sepharose 2B gel filtration from citrate-PRP into citrate free buffer (buffer-GFP). Response increases, reaches a maximum and decreases with increasing calcium and/or magnesium concentration. For either calcium or magnesium alone, increasing response is proportional to a rate coefficient and, through an apparent ion-platelet association constant, to the fraction of platelet critical sites bound to cation. With both ions present, bound magnesium appears to inhibit bound calcium in excess of that accounted for by competition and a lower rate coefficient for bound magnesium. With citrate present in buffer-GFP, apparent association constants increase, excess magnesium inhibition is present, but systems are path dependent. Initial conditions appear to establish a response which is thereafter immutable to environmental magnesium alteration. Citrate-PRP resembles buffer-GFP: response is sensitive to the selective removal of calcium and excess magnesium inhibition is present. With heparin-PRP, response is immutable to the selective removal of approximately 90% of initial calcium. The dependency of response inhibition observed at high divalent cation concentrations indicates that aggregation is not due to interplatelet cross linking by ions. Ion effects are similar for bovine and human platelets.

Adenosine Diphosphate↗
PubMed1975

The interactions of thrombin with borosilicate glass surfaces.

Borosilicate glass (G) and commercial poly(methyl methacrylate) (PMM) surfaces were examined at pH 7, monovalent ionic strength 0.14 and 22 degrees C. Thrombin concentrations ranged from 6 X 10(-9) to 5.2 X 10(-8)M. Decreases in thrombin concentration in 10 ml vessles were determined by clotting time assay. Adsorption to PMM vessels is small. These were examined to establish the validity of the method. For G-vessels, the terminations of initial rapid decreases, associated with adsorption equilibrium, are accounted for mainly by an average isotherm having an association constant of 1.8 X 10(7) l./mol. Adsorption equilibrium is accompanied by a slower rate of decrease consistent with the surface denaturation of 2.5%/min of adsorbed molecules. Molecules remaining adsorbed to the surfaces of 1 mm i.d. G-capillaries were examined using the sequence of thrombin adsorption, buffer washing for times tw, removal of buffer and introduction of a fibrinogen aliquot for determination of effective surface thrombin concentration by a capillary clotting time. Most adsorbed molecules (less than 90%) essentially desorb within tw = 2 min, a second class (greater than 8%) desorb in tw similar to 30 min, and members of additional classes are still present after tw = 1440 min. For all of the properties examined, there is a small variance between vessels taken at the same time from the same batch, and a larger variance between groups of vessels taken at different times from the same batch or from different batches.

Adsorption↗