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Biomedical subjects

M W Hatton

Publications and source records attributed to M W Hatton.

At least 37 records · Page 2Linked to original sources
PubMed1994

Catabolism of plasminogen glycoforms I and II in rabbits: relationship to plasminogen synthesis by the rabbit liver in vitro.

The metabolisms of the two glycoforms of rabbit plasminogen have been compared in rabbits. Plasminogen I and II (ratio in plasma, 1:2.2) differ only in glycan content: plasminogen I probably possesses one N-glycan and one O-glycan, and plasminogen II only one O-glycan. New Zealand White (NZW) rabbits were injected intravenously with 125I-plasminogen I and 131I-plasminogen II, and blood samples were taken at regular intervals over 5 days. Kinetic behaviors were determined from protein-bound radioactivities using a three-compartment model. Fractional catabolic rates for plasminogen II in the vascular space (2.42 d-1) and the total body (0.56 d-1) were significantly greater than those measured for plasminogen I (1.12 and 0.45 d-1); half-lives were 1.53 and 1.23 days for plasminogen I and II, respectively (P < .01). Fractional distributions among the vascular, noncirculating vascular, and extravascular compartments were 0.41, 0.13, and 0.46 for plasminogen I, and 0.23, 0.11, and 0.65 for plasminogen II. From these data, we determined that plasminogen II was catabolized 4.8 times more rapidly than plasminogen I and was quantitatively contained largely in the extravascular space. By comparison, perfusion of rabbit livers ex corpora showed that plasminogen II was synthesized and released 5.0 times faster than plasminogen I over a 5-hour period. The possible roles for these glycoforms in vivo with respect to their different turnover rates and compartmental distributions are discussed.

Animals↗
PubMed1994

Displacement of fibrin-bound thrombin by r-hirudin precludes the use of 131I-r-hirudin for detecting pulmonary emboli in the rabbit.

Pulmonary emboli are detectable by filling defects in the pulmonary vasculature upon pulmonary angiography. Emboli derived from venous thrombi are rich in fibrin to which thrombin remains bound. Hirudin, a specific thrombin inhibitor, binds to thrombin to yield a 1:1 stoichiometric complex. We examined whether 131I-recombinant hirudin (r-hirudin) could be used to detect pulmonary emboli in rabbits. Clots were formed by re-calcifying rabbit plasma in vitro, and then injected (0.034 ml) into a femoral vein to lodge in the lungs. 131I-r-hirudin (29 +/- 4 microCi/kg) was injected intravenously but emboli could not be detected by gamma camera in real time. Post-mortem analysis of lung tissue showed that 131I-r-hirudin did not associate with emboli prepared with 125I-fibrin. Because of these findings, we used different techniques to look at the binding of hirudin to plasma clots. Clots formed in vitro were incubated with 131I-r-hirudin in the presence of equimolar amounts of 125I-albumin; specific binding of 131I-r-hirudin was not observed. Experiments with immobilized fibrin(ogen) showed that 125I-r-hirudin did not bind to and remain with fibrin-bound 131I-thrombin but did lead to the inactivation and displacement of up to 70% of bound thrombin as r-hirudin-thrombin complex; residual thrombin bound to fibrin remained active. Thus, released r-hirudin-thrombin complex is probably cleared rapidly from the region of the embolus in vivo; radioiodinated r-hirudin may not, therefore, be useful as a marker for detecting emboli.

Animals↗
PubMed1994

Radiolabeled r-hirudin as a measure of thrombin activity at, or within, the rabbit aorta wall in vitro and in vivo.

The behavior of 125I-labeled recombinant hirudin towards the uninjured and de-endothelialized rabbit aorta wall has been studied in vitro and in vivo to determine its usefulness as an indicator of thrombin activity associated with the aorta wall. Thrombin adsorbed to either sulfopropyl-Sephadex or heparin-Sepharose bound > 95% of 125I-r-hirudin and the complex remained bound to the matrix. Binding of 125I-r-hirudin to the exposed aorta subendothelium (intima-media) in vitro was increased substantially if the tissue was pre-treated with thrombin; the quantity of 125I-r-hirudin bound to the de-endothelialized intima-media (i.e. balloon-injured in vitro) correlated positively with the quantity of bound 131I-thrombin (p < 0.01). Aortas balloon-injured in vivo were measured for thrombin release from, and binding of 125I-r-hirudin to, the de-endothelialized intimal surface in vitro; 125I-r-hirudin binding correlated with the amount of active thrombin released (p < 0.001). Uptake of 125I-r-hirudin by the aorta wall in vivo was proportional to the uptake of 131I-fibrinogen (as an indicator of thrombin activity) before and after balloon injury. After 30 min in the circulation, specific 125I-r-hirudin binding to the uninjured and de-endothelialized (at 1.5 h after injury) aorta wall was equivalent to 3.4 (+/- 2.5) and 25.6 (+/- 18.1) fmol of thrombin/cm2 of intima-media, respectively. Possibly, only hirudin-accessible, glycosaminoglycan-bound thrombin is measured in this way.

Animals↗
PubMed1993

On glucose transport and non-enzymic glycation of proteins in vivo.

Non-enzymic glycation is a chance event which may occur whenever a protein is in solution with a reducing sugar. The product of this reaction is a covalently linked glycated protein. Plasma proteins are commonly targets of glycation, particularly in the hyperglycaemic circulation, and yet little is known of the changes to protein function caused by glycation or the catabolic fate of these glycated proteins in vivo. The following article examines the effect of glycation on the catabolism of albumin, the principal glycated protein in plasma, by comparing the catabolisms of radio-labelled samples of rabbit naturally glycated and unglycated albumins in normal and diabetic rabbits. We reason that the relatively small changes (5-10%) to the half-life (T1/2) of albumin and to its distribution to body compartments caused by glycation in vivo may reflect an evolutionary adaptation of albumin (and of all plasma proteins) to resist glycation and, possibly, an accelerated catabolism. Presumably, any increase in the rate of catabolism of a glycated protein followed by its de novo replacement with native (unglycated) protein would create an extra energy demand of the host. In contrast to the vertebrates, the use of non-reducing disaccharides as transport fuels in the circulation systems of insects and higher plants may be an adaptation to allow transport of relatively high concentrations of sugar without the problems caused by glycation.

Animals↗
PubMed1993

Transient morphological and biochemical alterations of arterial proteoglycan during early wound healing.

Proteoglycan alterations in the carotid arteries of normolipemic rabbits during the first 14 days after injury were evaluated by morphometric analysis of ruthenium red-stained sections using transmission electron microscopy and by incorporation of 35S-sulfate. Two types of de-endothelializing injury were compared, air-drying and balloon catheter. Three days after either injury, a transient increase in concentration of 35S-labeled glycosaminoglycans, changes in distribution of 35S-sulfate among glycosaminoglycans, and a two- to threefold increase in 35S-sulfate content (measured as 35S/micrograms glycosaminoglycan) were observed. Morphometric analysis revealed an alteration in medial proteoglycan distribution and morphology at 3 days after injury, which was more evident after air-drying than balloon injury. In response to either injury, metabolically activated smooth muscle cells were associated with very large proteoglycan granules (diameter: > 60 nm) which possibly contained glycosaminoglycans with longer chains and/or altered charge densities. By 14 days after either injury, the distribution of medial proteoglycan returned to normal and a neointima formed. The neointima was thicker following balloon injury, but irrespective of the nature of the injury proteoglycan concentration was higher in the re-endothelialized than in non-reendothelialized areas. The alterations in extracellular proteoglycan of the intima-medial layers induced by these two forms of injury may influence the pattern of wound healing but are not associated with lipid deposition within the time frame examined.

Air Movements↗
PubMed1993

Evidence for thrombin binding to dermatan sulphate sites in the rabbit aorta subendothelium in vitro.

The proposal that thrombin binds to dermatan sulphate chains of extracellular proteoglycans has been examined directly using the subendothelium of the rabbit aorta. Freshly excised aortas were de-endothelialized by balloon catheter in vitro and then incubated with 125I-thrombin to allow adsorption of 20-30 fmol of thrombin/cm2. Pretreatment of the subendothelium with FPR-thrombin or chondroitinase ABC partially inhibited thrombin binding, each by approximately 40-45%. The addition of dermatan sulphate inhibited, competitively, up to 50% of thrombin from binding to the subendothelium whereas chondroitin-4 or -6 sulphates had little or no effect. By contrast, protamine inhibited 90% of FPR-thrombin binding. Of subendothelium-bound thrombin, chondroitinase ABC released only a small proportion (3-12%) of bound thrombin but up to 44% of bound FPR-thrombin. It is concluded that, when 125I-thrombin is bound in vitro at a concentration of < 30 fmol/cm2 of aorta intima-media, approximately 50% of subendothelial 125I-thrombin is bound to dermatan sulphate chains of proteoglycan in the extracellular matrix. The possibility is discussed that dermatan sulphate chains may function as thrombin-binding loci to control or augment thrombin activity in the ECM of the injured vascular wall in vivo.

Animals↗
PubMed1993

Catabolism of unglycated and naturally glycated forms of rabbit fibrinogen: their interaction with the healthy and deendothelialized aorta wall in normal and diabetic rabbits.

The incidence of nonenzymatic glycation of proteins within the hyperglycemic environment of diabetic plasma is increased compared with that in normal (i.e., nondiabetic) plasma. Whether glycation in vivo alters the behavior of proteins within the circulation is not well understood. Glycated fibrinogen, although not detected in normal rabbit plasma, was isolated from diabetic rabbit plasmas (glucose concentration 12 to 39 mmol/L) and separated from unglycated fibrinogen by boronate chromatography. The yield of glycated fibrinogen, which amounted to 3% to 6% of total fibrinogen, correlated with the content of plasma glucose. Glycated and unglycated fibrinogens facilitated aggregation of normal or diabetic platelets to a similar extent after adding adenosine-5'-diphosphate. Normal platelets stimulated by adenosine-5'-diphosphate bound more iodine 131-glycated fibrinogen than iodine 125-unglycated fibrinogen (p < 0.05), whereas the quantities of glycated and unglycated fibrinogens bound by diabetic platelets were not significantly different. When coinjected intravenously into normal or diabetic rabbits, 131I-glycated fibrinogen was cleared from the circulation faster than 125I-unglycated fibrinogen although the catabolic rates, measured as half-life, were not significantly different. At equilibrium, glycated fibrinogen was distributed significantly more in the extravascular and less in the vascular compartments of the normal and diabetic rabbit compared with the unglycated type. After balloon injury to the aorta in vivo, the unglycated/glycated ratio of radiolabeled fibrinogens associated with the platelet monolayer was 0.94, whereas for the damaged subendothelium the ratio was 1.20 (p < 0.005). We conclude that glycation in vivo changes several metabolic characteristics of fibrinogen in the normal and diabetic rabbit.

Animals↗
PubMed1992

Molecular cloning and expression of rabbit antithrombin III.

A cDNA containing the complete open-reading frame encoding rabbit antithrombin III (AT-III) was isolated from a rabbit liver cDNA expression library, using a specific antibody as a probe. Sequence analysis showed 84% identity between the deduced amino acid sequences of the rabbit and human proteins. A previously described cell-free expression system was used to verify the identity of the clone. The full-length cDNA was inserted into an expression vector, and messenger RNA (mRNA) transcripts generated. In vitro translation of these transcripts, in the presence of [35S]methionine, in an mRNA-dependent rabbit reticulocyte lysate system resulted in the synthesis of a 51-Kd polypeptide, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This nonglycosylated protein was capable of forming SDS-stable complexes with human alpha-thrombin. Complex formation was significantly enhanced following the deletion of nucleotides encoding the signal peptide, and the resultant generation of a 47-Kd nonglycosylated mature protein product. When the template DNA giving rise to this product was internally truncated, two rabbit AT-III deletion mutants were generated that lacked the ability to interact with thrombin, but retained the ability to bind heparin. Cell-free expression plasmids encoding the human and rabbit AT-III mature molecules were manipulated to produce two interspecies fusion proteins. For the first, human codons were used to replace rabbit codons from residue 369-433, while in the second human codons replaced rabbit codons from residue 217-433. Both fusion proteins exhibited less efficient thrombin-complexing ability than the original cell-free-derived mature rabbit AT-III. Thus, portions of AT-III molecules from the two species, despite their high degree of homology, are not interchangeable. Knowledge of the structure of rabbit AT-III, combined with the availability of the rabbit cDNA, will permit defined experimentation aimed at understanding antithrombin III structure relative to its function in vivo.

Amino Acid Sequence↗
PubMed1992

Increased platelet, but unaltered fibrinogen, accumulation in experimental thrombi in alloxan-induced diabetic rabbits.

Platelets from diabetic humans and animals have been found previously to be hypersensitive to agonists, including thrombin, in vitro but it is unclear if this hypersensitivity also occurs in vivo and leads to a greater thrombotic tendency. In the present study, the effect of diabetes was examined on thrombus formation and vessel wall responses which result from continuous intimal injury induced by indwelling aortic catheters in rabbits. Platelet and fibrin(ogen) associated with the thrombus and damaged aortae were examined. Control or alloxan-induced diabetic rabbits (9-12 months after initial treatment) were injected with 51Cr-labeled autologous platelets and 125I-labeled fibrinogen (prepared from control rabbits) before insertion of indwelling aortic catheters. The anesthetized rabbits were perfused-fixed after 20 hr or 4 days. The dry weight of thrombus that formed was determined and platelet and fibrin(ogen) accumulation in thrombi and on injured aortae were calculated from the associated 51Cr and 125I, respectively. In diabetic rabbits, more platelets accumulated in the thrombi which formed after either 20 hr or 4 days, although the weight of thrombus and net fibrin(ogen) incorporation into the thrombus were not different from corresponding control rabbits. Net platelet and fibrin(ogen) association with the injured aortae were not different between control and diabetic rabbits. It is likely that the increased platelet accumulation in arterial thrombi in diabetic rabbits which results from continuous injury to aortae is a consequence of hypersensitivity of these platelets to thrombin generated in the thrombus and at the sites of vessel injury.

Alloxan↗
PubMed1992

Altered levels of antithrombin III and fibrinogen in the aortic wall of the alloxan-induced diabetic rabbit: evidence of a prothrombotic state.

The distribution and behavior of the rabbit plasma proteins albumin, fibrinogen, and antithrombin III (ATIII) (isoforms alpha and beta), have been examined in groups of alloxan-induced diabetic rabbits and control rabbits. By injecting radiolabeled preparations intravenously, measurements of plasma clearance, rates of catabolism, and compartmental distribution were made for each protein. In addition, after allowing the radiolabeled proteins to circulate for 12 hours, we excised aortas after exsanguination and determined the content of these proteins in the endothelium and subendothelium. The respective fractional catabolic rates of ATIII-alpha and ATIII-beta were similar in the diabetic and control rabbits, but fibrinogen and albumin were catabolized more slowly in the diabetic rabbit than in the control rabbit. The distributions of albumin and the ATIII isoforms between the intravascular, noncirculating vascular, and extravascular compartments in the diabetic rabbit were similar to the respective proteins in the control rabbit, but a smaller proportion of fibrinogen was associated with the vascular compartment of the diabetic rabbit when compared with that in the control rabbit. At 12 hours after injection, the quantities of fibrinogen and albumin associated with the diabetic aorta endothelium and particularly the subendothelium were increased, whereas ATIII-alpha and ATIII-beta were decreased relative to the control aorta. The fibrinogen-to-ATIII ratio in the diabetic aorta was increased twofold to threefold when compared with that in the control aorta. We conclude that the increased ratio of fibrinogen to ATIII in the aorta wall of the diabetic rabbit may be characteristic of the prothrombotic state that is conspicuous in insulin-dependent diabetes.

Albumins↗
PubMed1991

Comparison of the effects of heparin and hirudin on thrombin binding to the normal and the de-endothelialized rabbit aorta in vitro.

The properties of heparin and hirudin to inhibit thrombin from binding to the freshly-excised rabbit aorta wall were compared in vitro. When aorta segments were incubated with 125I-thrombin (4.4 +/- 0.4 nM) in the presence of heparin or hirudin, both anticoagulants inhibited 125I-thrombin binding to the endothelium in a concentration-dependent manner (IC50: 0.1 USP U heparin/ml; 0.1 ATU hirudin/ml). Endothelium-bound 125I-thrombin was displaced by either heparin (50% liberated at 4.1 U/ml) or hirudin (0.4 U/ml). Using de-endothelialized aortas, heparin inhibited thrombin binding by the exposed subendothelium (IC50: 1.8 U/ml) whereas hirudin was without effect. Neither heparin nor hirudin was able to significantly liberate thrombin bound to the exposed subendothelium. These observations suggest that both heparin and hirudin mask the binding site on thrombin to the endothelial cell membrane. A separate site on thrombin must bind to the subendothelium because only heparin inhibits binding. Thrombin, although bound reversibly to the endothelium, is bound irreversibly to the exposed subendothelium due, probably, to reaction with endogenous extracellular antithrombin activities (e.g. antithrombin-III, protease nexin-1).

Adsorption↗
PubMed1990

Comparative behavior of thrombin and an inactive derivative, FPR-thrombin, toward the rabbit vascular endothelium. Heparin liberates FPR-thrombin from the endothelium in vivo.

Thrombin rapidly binds to and saturates rabbit aorta endothelium in vitro, a process that depends on pericellular glycosaminoglycans and that is inhibited by heparin. To characterize the initial adsorption of thrombin to the endothelium in vivo, an enzymatically inactive derivative, FPR-thrombin (i.e., thrombin inactivated by D-Phe-Pro-Arg-chloromethyl ketone), was prepared. The binding characteristics of thrombin and FPR-thrombin to heparin-Sepharose and to the endothelial surface of rabbit aorta segments in vitro were compared. From these experiments, we concluded that FPR-thrombin mimicked, qualitatively, the binding of thrombin to the endothelium. When injected intravenously, 125I-FPR-thrombin was removed rapidly from the rabbit circulation (T1/2, approximately 1.4 minutes) and simultaneously was adsorbed by the vascular endothelium, particularly in the lung. By injecting heparin (1,000 units/kg i.v.) before 125I-FPR-thrombin, adsorption by the aorta endothelium at 30 minutes after injection was reduced by 90%, and T1/2 was increased to approximately 3.4 minutes. Heparin, administered at various times after 125I-FPR-thrombin, liberated a significant proportion of 125I-FPR-thrombin from the endothelial surface into the plasma compartment as shown by a pronounced "spike" on the plasma curve, a concomitant loss of radioactivity from the lung and from the aorta endothelium, and analysis of the radioactive components of plasma taken before and after heparin injection. Thus, FPR-thrombin was cleared rapidly from the circulation, and endothelium-bound FPR-thrombin was released into the circulation by heparin.

Amino Acid Chloromethyl Ketones↗
PubMed1990

Enhanced binding of fibrinogen by the subendothelium after treatment of the rabbit aorta with thrombin.

Damage to the endothelial surface of an artery invokes a hemostatic response that causes platelet deposition and activation of the coagulation and fibrinolytic pathways on the exposed subendothelial surface. Plasma fibrinogen is rapidly adsorbed at the site of injury. To gain insight into fibrinogen uptake, undamaged and de-endothelialized rabbit thoracic aortas were pretreated with various concentrations of thrombin and then incubated with fibrinogen labeled with iodine 125 in vitro. Uptake of fibrinogen by the subendothelium was not affected by low thrombin concentrations (less than 10 nmol/L), probably because of the antithrombin capacity of the vessel wall to inactivate any thrombin adsorbed. Over the thrombin concentration range of 10 to 90 nmol/L. fibrinogen binding increased linearly as binding of thrombin labeled with iodine 131 increased. In contrast, treatment of the subendothelium with enzymatically inactive thrombin did not enhance fibrinogen binding. Fibrinogen binding was inhibited by exposing the thrombin-treated subendothelium to hirudin or phenylalanyl-prolyl-arginyl-chloromethyl ketone. High thrombin concentrations (greater than 100 nmol/L) caused either a steadily decreasing uptake of fibrinogen with low fibrinogen concentrations or fibrin coagulation on the subendothelial surface from a high fibrinogen concentration. Glycyl-prolyl-arginyl-proline (0.1 mg/ml), a selective inhibitor of fibrin polymerization, inhibited 72% to 78% of fibrinogen uptake by the thrombin-treated subendothelium. Fibrinogen uptake was Ca2(+)-dependent, but ethylenediaminetetraacetic acid (10 mmol/L) did not displace subendothelium-bound fibrinogen. Plasmin effectively removed at least 75% of bound fibrinogen, indicating an extracellular location for the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Wound healing in the media of the normolipemic rabbit carotid artery injured by air drying or by balloon catheter de-endothelialization.

The response to air-dry injury to the carotid artery of the normolipemic rabbit was compared with the response to de-endothelialization with a balloon catheter. Air drying induced an inflammatory response that resembled arteritis rather than atherosclerosis. There was medial damage, neutrophil but not macrophage infiltration, and fibrin formation, limited smooth muscle proliferation, which regressed after 3 months, and no lipid deposition. Within 1 week the smooth muscle cells were mainly of the secretory phenotype, and a neointima had formed. At 4 weeks the neointimal proliferation continued, but most cells showed a contractile phenotype. By 3 months, the lesion consisted of fibromuscular thickening with few small smooth muscle cells. Balloon injury induced minimal medial damage and continuing intimal proliferation with no evidence of regression by 3 months. It is concluded that air drying the carotid artery induces smooth muscle damage as well as endothelial cell loss, and this stimulates a wound-healing mechanism that is different from the response to selective intimal injury.

Air↗
PubMed1989

Deendothelialization in vivo initiates a thrombogenic reaction at the rabbit aorta surface. Correlation of uptake of fibrinogen and antithrombin III with thrombin generation by the exposed subendothelium.

Purified radiolabeled fibrinogen and antithrombin III (ATIII) were injected intravenously into rabbits before a deendothelializing injury to the aorta, and allowed to circulate for 0.1 to 6 hours before exsanguination, excision of the aorta, and quantification of each protein/unit area of subendothelium (intima-media). Uptake of fibrinogen was rapid (saturation 10 minutes after injury was approximately 13.0 pmol/cm2) compared with that of ATIII (45 to 60 minutes; 3.5 to 4.3 pmol/cm2). Both proteins associated primarily (greater than 90%) with the subendothelium rather than the platelet monolayer. The avidity of the deendothelialized vessel of these proteins was measured after a 20-minute circulation time at various intervals after injury. Whereas turnover of fibrinogen was fairly constant (approximately 100% per hour), that of ATIII was maximal (approximately 200% per hour) at 1 hour, decreasing to approximately 105% per hour at 5 hours after injury. The profile of ATIII turnover mirrored that of thrombin released in vitro from the deendothelialized aorta up to 10 days after injury, whereas the uninjured aorta and the aorta deendothelialized ex vivo adsorbed fibrinogen poorly and released negligible thrombin. Pretreatment of the aorta, deendothelialized ex vivo with thrombin in vitro increased fibrinogen uptake significantly. It is possible that, after deendothelialization in vivo, fibrinogen adsorption is determined largely by thrombin generation at the vessel wall. ATIII binding is limited by the availability of binding sites in the subendothelium, although the rate of thrombin generation influences ATIII turnover.

Animals↗
PubMed1988

Extraction and analysis of glycosaminoglycans from intima-media of single rabbit aortae: effect of balloon catheter de-endothelialization on the content and profile of glycosaminoglycans.

A novel procedure is described for extracting, and simultaneously 3H-labelling, glycosaminoglycans from the intima-media of a single rabbit aorta. The procedure was used to compare contents and types of glycosaminoglycans isolated from uninjured (control) aortae, and partially re-endothelialized aortae at 11 weeks after de-endothelialization by a balloon catheter. Briefly, the isolated delipidated tissue was digested in 0.8 M NaOH containing NaB3H4 at room temperature. The neutralized digest was then degraded by a non-specific proteinase during dialysis. The 3H-labelled glycosaminoglycan fraction was recovered after a gel filtration step. The yield (10.5 micrograms of glycosaminoglycan/mg of dry, delipidated tissue) was within the range reported previously for rabbit aorta. Although the de-endothelialized (DEA) and re-endothelialized areas (REA) of all injured aortae contained a significantly thickened intima, the glycosaminoglycan concentration (DEA, 11.2 micrograms/mg; REA, 11.6 micrograms/mg) did not differ significantly from that of control aorta. The profile of [3H]glycosaminoglycan types was determined by the serial use of glycosaminoglycan-selective methods: greater than 86% of 3H was released as small molecular weight products by this analytical scheme. The glycosaminoglycan profile for control tissue compared well with several previous reports. Compared with control aortae, both DEA and REA contained relatively less chondroitin sulphate, whereas DEA contained more hyaluronic acid and REA contained more heparan sulphate. Future use of this procedure will improve measurement of the changes to the extracellular matrix which take place after injury to the vessel wall and which may precede atherogenesis.

Animals↗