Anticoagulant and Antithrombotic Properties of Three Structurally Correlated Sea Urchin Sulfated Glycans and Their Low-Molecular-Weight Derivatives.

The anticoagulant and antithrombotic properties of three structurally correlated sea urchin-derived 3-linked sulfated α-glycans and their low molecular-weight derivatives were screened comparatively through various in vitro and in vivo methods. These methods include activated partial thromboplastin time, the inhibitory activity of antithrombin over thrombin and factor Xa, venous antithrombosis, the inhibition of platelet aggregation, the activation of factor XII, and bleeding. While the 2-sulfated fucan from Strongylocentrotus franciscanus was observed to be poorly active in most assays, the 4-sulfated fucan from Lytechinus variegatus, the 2-sulfated galactan from Echinometra lucunter and their derivatives showed multiple effects. All marine compounds showed no capacity to activate factor XII and similar low bleeding tendencies regardless of the dose concentrations used to achieve the highest antithrombotic effect observed. The 2-sulfated galactan showed the best combination of results. Our work improves the background about the structure-function relationship of the marine sulfated glycans in anticoagulation and antithrombosis. Besides confirming the negative effect of the 2-sulfated fucose and the positive effect of the 2-sulfated galactose on anticoagulation in vitro, our results also demonstrate the importance of this set of structural requirements on antithrombosis in vivo, and further support the involvement of high-molecular weight and 4-sulfated fucose in both activities.

Intra-articular tissue factor/factor VII complex induces chronic arthritis.

OBJECTIVE: Fibrin accumulation in the joint cavity is a common feature of chronic arthritides, such as rheumatoid arthritis (RA). Complex formation between tissue factor (TF) and factor VII (FVII) is the initial step in such a fibrin formation. METHODS: To assess the role of the TF/FVII complex in the pathogenesis of joint inflammation, 1) the levels of TF/FVII complex were measured in synovial fluid of RA patients; 2) the complex was injected to healthy mice intra-articularly. RESULTS: Morphological analysis of the joints 4 days after TF/FVII injection revealed influx of CD4-Mac1+ mononuclear leukocytes into synovial tissue followed by cartilage and bone destruction. Inflammation induced by TF/FVII complex was more profound than that caused by each of the proteins separately, both with respect to frequency, severity and duration of arthritis. Interaction between macrophages and lymphocytes in sustaining joint inflammation was proved by the requirement of the combined lymphocyte/ monocyte depletion to abolish TF/FVII induced arthritis. Induction of monocyte attracting chemokines (MIP-1 alpha and RANTES) was shown to be one of the potential mechanisms for TF/FVII complex triggered inflammatory cell influx. Interestingly, TF/FVII complexes were detected in synovial fluid of 20/40 patients with RA. CONCLUSIONS: Altogether these findings indicate that TF/FVII complexes, frequently found intra-articularly in joints of RA patients, may be an important component in both induction and progression of chronic destructive arthritis.

[Protective effect of heparin and phosphatidylserine in exogenous thromboplastinemia]. / Zashchitnyi éffekt geparina i fosfatidilserina pri ékzogennoi tromboplastinemii.

It was shown in experiments on rats that heparin and phosphatidylserine-containing anticoagulant increase the animals' tolerance to thromboplastin administration to the blood flow. This is manifested by a considerable lowering of the death rate and restriction of fibrinogen consumption caused by thromboplastin. It was ascertained that the protective effects of heparin and phosphatidylserine are drastically potentiated during combined administration, apparently because of the influence of the anticoagulants on different stages of fibrin formation.

Properties of canine tissue thromboplastins from brain, lung, arteries, and veins.

Properties of the protein moieties of canine tissue thromboplastins (TTP's) from brain (BTTP), lung (LTTP), arteries (ATTP), AND VEINS (VTTP) were determined. The maximum specific activity of each protein moiety after its relipidation was obtained when the phospholipid-delipidated TTP ratio was 0.32 and was 1,395 U/mg BTTP, 1,130 U/mg LTTP, 630 U/mg VTTP, and 435 U/mg ATTP. The amino acid contents of the protein moieties of LTTP, ATTP, and VTTP were closely similar, but that of BTTP was significantly different. The Ouchterlony analysis showed that BTTP did not react at all with the antibody against VTTP, but that three other TTP's did and showed the reaction of complete identity. Then, the reactivity of 125-I-labeled TTP's with the anti-VTTP antibody was studied. The results showed that 0.79 +/- 0.01 (SD) % of [125I]BTTP, 10.24 +/- 0.5 (SD) % of [125I]LTTP, 19.4 +/- 0.2 (SD) % of [125I]VTTP, and 5.88 +/- 0.4 (SD) % of [125I]ATTP added were bound to the antibody in 2 h. Next, the molecular weight of each was determined by Sephadex G-200 filtration, which averaged 80,000 +/- 4,000 (SD) ([125I]BTTP), 113,000 +/- 5,000 (SD) ([125I]LTTP), 62,000 +/- 3,000 (SD) ([125I]ATTP), and 47,000 +/- 2,000 (SD) ([125I]VTTP). Finally, the plasma behavior of each was studied in four dogs. The plasma half-life averaged 8.1 +/- 0.24 (SD) h ([125I]BTTP), 14.6 +/- 0.5 (SD) h ([125I]LTTP), 7.38 +/- 0.48 (SD) h ([1252]ATTP), and 24.3 +/- 0.9 (SD) h ([125I]VTTP). These results indicate that the protein moieties of canine TTP's from brain, lung, arteries, and veins are closely similar in some aspects but dissimilar in others and are definitely not identical.