The problem is discussed of providing a theoretical explanation for the empirical equation of Laurent, Bjork, Pietruszkiewicz & Persson (1963) which relates the relative retardation of the sedimentation of compact particles in solutions of hyaluronic acid to the radius of the particle and the concentration of hyaluronic acid. From published and original data, the same relation is shown to apply also to sedimentation and diffusion in solutions of a number of linear and branched chain-polymers. Of several approaches to the problem, only one, based on the stochastic model of diffusional migration, yields the empirical relationship, and predicts a value of the numerical constant close to that observed. This treatment is shown to apply to other forms of migration, including the case where the chain-polymer is itself migrating. The theory is tested by calculation, from migration data, of the effective radii of the polymer chains. The results are consistent and comparable with values deduced from equilibrium experiments.

1. Materials containing hyaluronic acid have been prepared by filtration (Ogston & Stanier, 1950) from ox synovial fluid and from a protein-rich human mesothelioma fluid. The ox material has been deproteinized by treatment with chloroform and pentanol and by gradient elution on DEAE-Sephadex; several fractions were obtained by the latter method. These materials can be stored in solution at -20 degrees without change of properties. The ox material contained 21% of protein; all other preparations contained less than 6% of protein. 2. The two materials have been compared by sedimentation and viscosity and shown to be closely similar. Treatment of the ox material with neuraminidase caused no change in its viscosity behaviour. 3. Information about the molecular configuration of the ox material has been obtained from measurements of light-scattering and viscosity. The results, though consistent with a highly extended configuration, are not consistent with a linear random-coil configuration. It is tentatively suggested that the structure may have some degree of branching and of cross-linking, which give it a rigidity with respect to expansion of the molecular domain that would not be possessed by a random coil. 4. The deproteinized material recovered from DEAE-Sephadex, though polydisperse, showed unchanged average molecular weight; however, the average radius of gyration was greater than before this treatment. 5. Acidification to approx. pH3 resulted in a contraction of the structure, with only a slight degree of expansion when the pH was restored to 6.8-7.0. 6. Measurements of optical rotatory dispersion qualitatively support a structure less simple than a linear random coil. 7. Colloid osmotic pressures of mixed solutions of bovine serum albumin and of hyaluronic acid prepared by filtration from ox synovial fluid have been measured. The results agree approximately with those of Laurent & Ogston (1963) but are in quantitative disagreement with the partition measurements of Ogston & Phelps (1960). The relationships between thermodynamic quantities in a quaternary system of electrolytes are discussed in Appendix 2. 8. Refractometric measurements have been made in connexion with light-scattering measurements, as the basis for a convenient method of determining the concentrations of solutions of hyaluronic acids, and to measure the partition of sodium chloride in dialysis experiments. The theory of the last use is discussed in Appendix 1. 9. Sedimentation measurements on the ox preparation have been made up to a concentration of 1.4x10(-2)g./ml. The form of the sedimentation coefficient-concentration relationship is discussed. The value of the sedimentation coefficient at higher concentration is the basis of an illustration of the likely effect of hyaluronic acid on the flow of water through narrow channels in connective tissue. 10. Available colorimetric methods have been shown to give low estimates for glucuronic acid when applied to highly polymerized materials, as compared with estimates by decarboxylation. A spectrop...

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synopsisDonnan equilibria between solutions of various carbohydrate polyions and Nas have been measured by equilibrium dialysis. From the results, estimates have been made of the degree of Na@ fixation by t,he polyions. Comparison is made between the behavior of the complex proteoglycan macromolecules and their constituent glycosaminoglycans. Attempts to interpret these results and the Nae activity in saltfree solutions on the parameter of the intercharge distance has been made. By use of studies on carboxymethylcellulose, the relative importance of the carboxyl and of the sulphate ester groups in the Nae fixation is assessed. The relevance of the measurements to the previously described diffusional effects found in real and model connective tissues is discussed.