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Biomedical subjects

M L Tiffany

Publications and source records attributed to M L Tiffany.

8 recordsLinked to original sources

Technical considerations for platelet aggregation and related problems.

Platelet aggregation generally is ordered by the physician to evaluate platelet function in hemorrhagic or thrombotic disorders. Malfunction of the platelet may be the result of an intrinsic congenital defect or an acquired problem induced by drugs or certain circulating plasma factors. It is necessary to obtain information from the patient with respect to family history, drug ingestion, physical or mental stress. In addition, other laboratory studies should be obtained to rule out general coagulation disorders affecting the plasma factors. A bleeding time will be helpful in establishing the severity of any platelet dysfunction. Technical considerations with regard to the preparation of the samples are of primary importance in determining platelet aggregation. Aggregating studies require the use of a variety of binding agents. (Studies on shape change, adhesion of platelets, release of platelet granule substance, and or lysis with extrusion of cytoplasmic constituents may be helpful in certain cases.) Instrumentation for platelet aggregation presently is available in many hospitals. The technical factors to be considered for routine aggregation studies include the type and strength of anticoagulant, centrifugation technique used in preparing the platelet-rich and platelet-poor plasma, platelet concentration, time of storage of the sample after venipuncture and after centrifugation, temperature, and the mixing of the sample. In general, critical concentrations of each reagent should be employed to improve the discrimination capability of the assay. Small differences in response may be obliterated by using excessive concentrations of a given reagent. Comparison in response to the test platelets with control platelets is best done at the same time by performing the aggregation in a dual instrument so that handling procedures will be identical and artifactual differences eliminated.

Aging

Effect of complement on collagen-induced platelet aggregation.

Evidence that early members of the classic pathway of complement are involved in the interaction of collagen with the blood platelet is presented. C4 is required for platelet aggregation response to low concentrations of fibrous collagen but not for adhesion of collagen to platelets obtained from guinea pigs genetically lacking C4. The aggregation response is restored, however, by preincubation with either C4 or normal plasma. It is suggested that membrane-bound C1s is the receptor site for collagen, inasmuch as preincubation of normal platelets with antiserum to C1q specifically enhances the platelet-collagen interaction, demonstrating a potential competition between C1q and collagen for the platelet binding site. This concept is further supported by the fact that C1s inhibitors also enhance aggregation response to collagen. Under physiologic conditions, the role of complement in the platelet response to collagen should be highly significant.

Animals

A circular dichroism study of charged polypeptides interaction with salts.

The effect of salts on the experimental circular dichroism spectra of polypeptides is presented using poly-L-lysine as the main model. Salt effects are analyzed into: (a) shielding at low (less than 0.5 M) concentrations of all salts; (b) binding to positively charged and some neutrally charged side-chains by certain anions (e.g., CCl3COO-, CF3C00-, ClO4-), with induction of helicity; (c) binding of these same anions, at high concentration, to the backbone leading toward random structure; (d) binding of high concentration of denaturing cations (La+3, Ca++, Li+) to the backbone, with La+3 and Ca++ leading to collapsed random structure (R) while Li+ tends to leave the polypeptide somewhat extended; (e) indirect interaction of salting-out salts (NaH2PO4, (NH4)2SO4, NH4F), at high concentration, leading toward complete alpha helicity, probably by competition with the polypeptide and the anion for available water. Effects of changing the temperature from 5 degrees to 50 degrees on the circular dishroism spectra of different polypeptide-salt solutions throughout the region from extended (LES) to alpha helical conformation are analyzed in terms of introduction of randomness (R) at high temperature. Applications to effects of salt on protein structures are considered.

Binding Sites