Increased Urinary β-Thromboglobulin Excretion in Diabetes Assayed with a Modified RIA Kit-Technique

Summary

A modification of the commercial radioimmunoassay (RIA) kit adapting to the assay of β-thromboglobulin (β-TG) in the urine, is described. The urine was concentrated by dialysis against dry Sepharose G 200. ^l31I-albumin was used as marker. Experiments in which ^l25I-β-TG and ¹³¹I-albumin were added simultaneously showed a parallel recovery of both markers in the final sample. The sensitivity of the RIA was enhanced by incubation overnight of the urine sample with the antiserum. Increased urinary pTG values were found in 20 patients with diabetes mellitus as compared to 20 young healthy normals. Increased urinary β-TG values correlated with decreased creatinine clearance, but the difference between diabetics and normals was still observed when patients with a creatinine clearance of less than 100 ml/min were excluded. The urine βTG level was correlated with the presence of neovascularization within the diabetic group.

Keywords

β-thromboglobulin - Diabetes - Microangiopathy - Radioimmunoassay

• References

• 1 Moore S, Pepper DS, Cash JD. The isolation and characterisation of platelet-specific β-globulin (β-thromboglobulin) and the detection of anti-urokinase and antiplasmin released from thrombin-aggregated washed human platelets. Biochim Biophys Acta 1975; 379: 360-369
• 2 Bolton AE, Ludlam CA, Moore S, Pepper DS, Cash JD. Three Approaches to the Radioimmunoassay of Human β-Thromboglobulin. Br J Haematol 1976; 33: 233-238
• 3 Ludlam CA, Bolton AE, Moore S, Cash JD. New rapid method for diagnosis of deep venous thrombosis. Lancet 1975; 02: 259-26
• 4 Kaplan KL, Nossel HL, Drillings M, Lesznik G. Radioimmunoassay of Platelet Factor 4 and β-Thromboglobul:Development and Application to Studies of Platelet Release in Relation to Fibrinopeptide A Generation. Br J Haematol 1978; 39: 129-146
• 5 Zahavi J, Kakkar VV. β-Thromboglobulin-a Specific Marker of In-Vivo Platelet Release Reaction. Thromb Haemostas 1980; 44: 23-29
• 6 Kaplan K. p-Thromboglobulin. Progr Hemost Thromb 1980; 05: 153-178
• 7 Ludlam CA, Cash JD. Studies on the Liberation of β-Thromboglobulin from human platelets in vitro. Br J Haematol 1976; 33: 239-247
• 8 Bastl CP, Musial J, Kloczewiak M, Guzzo J, Berman I, Niewiarowski S. Role of Kidney in the Catabolic Clearance of Human Platelet Antiheparin Proteins from Rat Circulation. Blood 1981; 57: 233-238
• 9 Dawes J, Smith RC, Pepper DS. The release, distribution and clearance of human P-thromboglobulin and platelet factor 4. Thromb Res 1978; 12: 851-861
• 10 Snedecor GW, Cochran WG. Statistical Methods. Iowa State University Press; Ames, Iowa: 1967. 6. th ed: Chapters 5 & 8
• 11 Begg GS, Pepper DS, Chesterman CN, Morgan FJ. Complete Covalent Structure of Human β-Thromboglobulin. Biochemistry 1978; 17: 1739-1744
• 12 Sander HJ, Slot JW, Bouma BN, Bolhuis PA, Pepper DS, Sixma JJ. Immunocytochemical localization of fibrinogen, platelet factor 4 and α-thromboglobulin in ultrathin cryostat sections of human blood platelets. J Clin Invest. 1983 in press
• 13 Ludlam CA, Moore S, Bolton AE, Pepper DS, Cash JD. The release of a Human Platelet Specific Protein measured by a Radioimmunoassay. Thromb Res 1975; 06: 543-548
• 14 Kaplan KL, Broekman MJ, Chemoff A, Lesznik G, Drillings M. Platelet α-Granule Proteins: Studies on Release and Subcellular Localization. Blood 1979; 53: 604-618
• 15 Musial J, Niewiarowski S, Edmunds LH, Addonizio VP, Nicolaou KC, Colman RW. In Vivo Release and Turnover of Secreted Platelet Antiheparin Proteins in Rhesus Monkey (Macaca Mulatta). Blood 1980; 56: 596-603
• 16 Depperman D, Andrassy K, Seelig H, Ritz E, Post D. Betathromboglobulin is elevated in renal failure without thrombosis. Thromb Res 1980; 17: 63-69
• 17 Bolton AE, Cooke ED, Lekwani CP, Bowcock SA. Urinary α-thromboglobulin levels as a diagnostic marker for post-operative deep vein thrombosis. Thromb Res 1980; 19: 249-255
• 18 Guzzo J, Niewiarowski S, Musial J, Bastl C, Grossman R A, Rao AK, Berman I, Paul D. Secreted platelet proteins with antiheparin and mitogenic activities in chronic renal failure. J Lab Clin Med 1980; 96: 102-113
• 19 Anderton JL, Fananapazir J, Dawes J. Urinary α-Thromboglobulin in Essential Hypertension. Br J Haematol 1980; 44: 307-311
• 20 Snedecor JW. Queries. Biometrics 1952; 08: 85-88