Serial Dilution Protamine Sulfate Test Research Articles

Function of phagocytes in experimentally induced colitis in rabbits. II. The Shwartzman phenomenon in the colon.

Adhesion, phagocytosis, chemotactic and random migration, nitroblue tetrazolium dye reduction of peritoneal exudate neutrophils and macrophages, fibrinogen level, gelation of soluble fibrin and serial dilution protamine sulfate test were investigated in 115 New Zealand white rabbits with experimentally induced Shwartzman phenomenon in the colon, and in control animals. The results presented in this report demonstrated impairment of chemotactic migration of phagocytes in the presence of endotoxin. The depression was dose-dependent and less marked when neutrophils were stimulated with monocyte-derived chemotactic factor or with casein, than with complement-derived chemotactic factor. The prolonged depression of chemotactic migration of neutrophils and macrophages in rabbits with colitis, however, did not affect the healing time of the ulcers in the colon.

Disseminated Intravascular Coagulation in Ponies with Surgically Induced Strangulation Obstruction of the Small Intestine

Total strangulation obstruction (S/O) of the jejuno‐ileum was produced in a group of randomly selected ponies. Peripheral blood samples were collected prior to surgery and at 3‐hour intervals following obstruction. A coagulation profile consisting of platelet count, fibrinogen titer (FT), prothrombin time (PT), activated partial thromboplastin time (APTT), and serial dilution protamine sulfate test (SDPS) for fibrin monomers (FM) and/or early fibrin degradation products (fdp) was evaluated. The packed cell volume (PCV) and total plasma protein (TPP) were also determined. Concurrent peritoneal fluid samples were assessed grossly. Sham‐operated (sham) ponies were treated in a similar manner, excluding the production of a strangulation obstruction of the small intestine. Notable coagulopathies occurred in the S/O group. PT and APTT were increased significantly in samples collected within 6 hours of death. All S/O horses developed strongly positive SDPS reactions, while sham ponies, presurgical S/O ponies, and early S/O postsurgical samples were unremarkable. There were no significant shifts in the fibrinogen titer. Platelet counts were significantly decreased during the final 10% of sampling time in S/O horses. The coagulopathies observed in the S/O ponies appear to be secondary to the pathophysiologic changes produced by the S/O and not related to surgical trauma.

Correlation of the Serial-dilution Protamine Sulfate Test with Models of “Hypercoagulability” and Thrombosis in Dogs

In a dog model, combinations of venous-injury thrombosis, homologous serum-induced "hypercoagulability," and fibrinolytic blockade were evaluated to determine the factors involved in the generation of a positive serial-dilution protamine sulfate test. Stasis and operative procedures did not produce a positive test. Small local thrombi induced by electrical current produced a positive test that tended to persist only during the initial formation and propagation of the thrombus, and was blocked by pretreatment with epsilon-aminocaproic acid, suggesting in this model that the test was detecting mainly early fibrin degradation products. Homologous serum infusion without venous-injury thrombosis also produced positive serial-dilution protamine sulfate test results that could not be blocked with epsilon-aminocaproic acid suggesting here that the serial-dilution protamine sulfate test was detecting mainly soluble complexes of fibrin monomer rather than early fibrin degradation products. The serial-dilution protamine sulfate test is therefore capable of detecting both small local thrombi and serum-induced "hypercoagulability" without thrombosis in the dog model.

Noninvasive Diagnosis of Deep Venous Thrombosis

Eighty-five patients suspected of having lower-extremity deep venous thrombosis (DVT) participated in a prospective study to test the diagnostic accuracy of four noninvasive techniques: Doppler ultrasonic flow study, electrical impedance plethysmography, the serial dilution protamine sulfate test, and an extensive physical examination. Ascending radiocontrast phlebography was the diagnostic standard of reference. We found that (1) when both Doppler and impedance examinations were positive, the diagnosis of DVT could be considered virtually certain; (2) impedance and Doppler examinations, when used in combination, were reliable screening tests capable of establishing or excluding the presence of thigh DVT; (3) physical examination and the serial dilution protamine sulfate test were unreliable screening techniques for DVT; (4) techniques other than the noninvasive methods investigated were needed to reliably detect or to exclude popliteal and call DVT.

Semiquantitative Determination of Soluble Fibrin Monomer Complexes by Chromatography and Serial-dilution Protamine Sulfate Test

Plasmas were prepared from blood obtained from rabbits and from healthy individuals and patients with thrombotic disorders. Gel filtration (Sepharose-4B) chromatography and the serial-dilution protamine sulfate (SDPS) test were performed on each sample. For healthy individuals and rabbits, a narrow fibrinogen peak was obtained on chromatogram and was associted with a negative SDPS test. After administration of thrombin or endotoxin to rabbits and in plasmas from patients with intravascular coagulation, major shifts to earlier effluent volumes occurred. These higher-molecular-weight derivatives corresponded to soluble fibrin monomer complexes (FM). Positive SDPS tests were found in these plasmas. There was a highly significant correlation between percentage area occupied by FM on chromatogram and SDPS test titer. It is concluded that the SDPS test is a reliable semiquantitative measure of FM that can be used clinically as well as in experimental animal models.

A Study of the Large Heparin Requirements in the Generalized Shwartzman Reaction

AbstractThe heparin requirements necessary to inhibit intravascular fibrin deposition and soluble fibrin monomer (FM) formation in the generalized Shwartzman reaction (GSR) were evaluated. Fibrin deposition was measured by a quantitative technique utilizing 125I-labeled rabbit fibrinogen. FM was measured semiquantitatively by gel exclusion chromatography and by the serial dilution protamine sulfate (SDPS) test. There was a fourfold increase in heparin requirement 5 min after compared with 5 min before the second dose of endotoxin. This increase in heparin requirement was not related to thrombin elaboration, since FM was not found until more than 1 hr after the second dose of endotoxin. Neither was there any evidence of diminished sensitivity to the anticoagulant effect of heparin. The heparin requirements in the GSR rabbits were found to be in excess of those needed to neutralize a defibrinating dose of thrombin. It was concluded that a potent, heparin-resistant clotting activity developed within 5 min of the second endotoxin injection. The mechanism by which the activity caused the gradual elaboration of a thrombin-like enzyme is difficult to explain on the basis of traditional coagulation reactions. The apparent role of white cells is discussed.

A study of the large heparin requirements in the generalized Shwartzman reaction

The heparin requirements necessary to inhibit intravascular fibrin deposition and soluble fibrin monomer (FM) formation in the generalized Shwartzman reaction (GSR) were evaluated. Fibrin deposition was measured by a quantitative technique utilizing 125I-labeled rabbit fibrinogen. FM was measured semiquantitatively by gel exclusion chromatography and by the serial dilution protamine sulfate (SDPS) test. There was a fourfold increase in heparin requirement 5 min after compared with 5 min before the second dose of endotoxin. This increase in heparin requirement was not related to thrombin elaboration, since FM was not found until more than 1 hr after the second dose of endotoxin. Neither was there any evidence of diminished sensitivity to the anticoagulant effect of heparin. The heparin requirements in the GSR rabbits were found to be in excess of those needed to neutralize a defibrinating dose of thrombin. It was concluded that a potent, heparin- resistant clotting activity developed within 5 min of the second endotoxin injection. The mechanism by which the activity caused the gradual elaboration of a thrombin-like enzyme is diffucult to explain on the basis of traditional coagulation reactions. The apparent role of white cells is discussed.

Identification of FgDP and FDP by serial dilution protamine sulfate test (SDPST)

The identification of FDP, fibrin monomer and FgDP is very important for clinical implication, so that, various methods have been reported. In recent years, the differentiation of intravascular coagulation from primary fibrinolysis has been performed by the laboratory technique based on the paracoagulation phenomena. The fundamental mechanism of paracoagulation is not yet entirely clear, but it may be true that the acidic peptides —split from fibrinogen by thrombin— are related to the phenomenon.In this paper, the authors made a fundamental study on the serial dilution protamine sulfate test (SDPST) reported by Niewiarowski and Gurewich (1971). According to our observations, the test is unaffected by the addition of various amounts of fibrinogen into normal plasma, and is also insensitive to high levels of FgDP added to norml plasma. But, FDP produced in both early and late stages of fibrinolysis gives a positive result on SDPST, even with 5μg/ml of early FDP. Showing a positive result on SDPST by late FDP in normal plasma is not in agreement with the original report. This may be due to residual thrombin activity in reaction mixtures. Fibrin monomer in normal plasma gives a positive result on SDPST. In conclusion, the SDPST assay is sensitive to only FDP and fibrin monomer, and therefore, the positive result of the test indicates the presence of preceded thrombin in vivo. Some clinical data of SDPST is presented and discussed in this paper.

Intravascular coagulation in obstetrics: Serial dilution protamine sulfate test throughout labor

Serial dilution protamine sulfate tests (SDPS) were performed in 191 women during labor. It was found that 27 per cent of women with uncomplicated pregnancies and 42 per cent with complicated pregnancies had positive SDPS tests, the highest incidence being between three and six hours of labor. These findings support the previous report of a physiologic disseminated intravascular coagulation in pregnancy but make the SDPS test of little value in obstetric cases.

フィブリン・フィブリノーゲンと動脈硬化

Fibrinogen derivatives in the cryoprecipitable portion of Blombäck's fraction I-O from 50 patients, mainly with atherosclerosis, were studied by SDS-polyacrylamide gel electrophoresis. No discernible stabilized fibrinogen-fibrin complex could be detected in the electrophoresis. Although stable fibrinogen dimer, probably made by disulfide linkage, were detected in all cases of atherosclerosis and also in normal controls, fibrinogen dimers having γ-γ diads could be observed in only two cases; one with acute myocardial infarction one day prior to the study and the other five months prior to the study. Fraction I-8, clottable derivatives of fibrinogen degradation, could also be found in all cases and in controls. Serial dilution protamine sulfate test showed negative for all plasma samples except one which had acute myocardial infarction one day prior to the study and showed γ-γ diads in the electrophoresis.Plasma of the atherosclerotic patients showed the increments of plasma fibrinogen concentration and fibrin degradation products together with the shortening of the euglobulin lysis time. Concentration of the plasma fibrin stabilizing factor had no relation to the aging nor to the plasma fibrinogen concentration. Levels of plasma fibrin stabilizing factor in two cases with γ-γ diads remained normal.Plasma fibrinogen could suppress both the polymerization and crosslinking of fibrin produced in the circulation. Increased plasma fibrinogen may accelerate the suppression of the polymerization and crosslinking of fibrin produced more under the hypercoagulable state of sclerotic patients.Possible roles of fibrinogen derivatives in the mechanisms of the deposition of insoluble fibrinogen derivatives in the aortic intima (by Shainoff, J. R.) were discussed.

Thrombolysis with Streptokinase in Rabbits Dose Response, Fibrin-clot Specificity and Laboratory Evaluation of Fibrinolytic Effect

SummaryAn animal model was used in which venous thromboemboli of reproducible size could be formed. The effects of SK 500–50,000 units per hour were evaluated and compared to those of saline. Proportionately the greatest clot lysis occurred with the lowest doses of SK. In contrast to thrombolysis, significant fibrinogenolysis did not occur in any of the animals indicating a high degree of specificity of SK-induced activator for rabbit fibrin. An excellent correlation between the concentration of fibrin degradation products (fdp) measured by the serial dilution protamine sulfate test and the extent of clot lysis was found suggesting this to be a useful method for monitoring the effects of thrombolytic therapy. A method for the laboratory distinction of fdp from fibrin monomer was demonstrated.

The laboratory diagnosis of venous thromboembolic disease by measurement of fibrinogen-fibrin degradation products and fibrin monomer.

The utility of two blood tests for screening patients with suspected venous thromboembolic disease was evaluated. Eighty-two patients who underwent venography and 38 patients who were investigated with radioisotope lung scans or pulmonary angiograms were included. At the time of their radiographic study, their blood was tested for fibrinogen/fibrin degradation products and fibrin monomer by the staphylococcal clumping test (SCT) and the serial dilution protamine sulfate (SDPS) test. Patients in whom the diagnosis of pulmonary embolism or symptomatic deep vein thrombosis was confirmed had a significantly higher incidence as well as more strongly positive blood tests compared to the remaining patients. The SCT was more sensitive but the SDPS test was found to be more specific. The study indicates that these tests are sufficiently practical and precise to be used to help resolve the differential diagnosis in suspected venous thromboembolism. When both tests were negative, significant thromboembolic disease was not found. Evidence is presented suggesting that the coagulation products measured represent the result and not the cause of the thrombosis.

Detection of intravascular coagulation by a serial-dilution protamine sulfate test.

A new protamine sulfate test, using serial dilutions of 1% protamine sulfate in tris buffer, is specific for two fibrinogen derivatives that indicate thrombin generation, fibrin monomers, and fibrin degradation products. It is sensitive to low levels of each. It is insensitive to fibrinogen or fibrinogen degradation products and thus readily distinguishes primary from secondary fibrinolysis. The test is not affected by therapeutic levels of heparin. In clinical experience with this test and the staphylococcal clumping test for fibrinogen-fibrin degradation products no false negative reactions have been encountered in disseminated intravascular coagulation or in acute pulmonary thromboembolism. In 85% of patients with acute deep vein thrombosis the protamine sulfate test was positive. Test results in normal subjects and in patients with a variety of conditions are presented here. The protamine sulfate test is a practical, specific, and sensitive blood test for the exclusion of venous thromboembolic disease and disseminated intravascular coagulation.

The Detection of Intravascular Coagulation by a Serial Dilution Protamine Sulfate Test.

Meeting Abstracts1 May 1971The Detection of Intravascular Coagulation by a Serial Dilution Protamine Sulfate Test.V. Gurewich, M.D., F.A.C.P., S. Niewiarowski, M.D., E. Hutchinson, B.S., J. Gajewski, M.D.V. Gurewich, M.D., F.A.C.P.Search for more papers by this author, S. Niewiarowski, M.D.Search for more papers by this author, E. Hutchinson, B.S.Search for more papers by this author, J. Gajewski, M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-74-5-826_4 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptIn vitro studies demonstrated that when serial dilutions of a 1% stock solution of protamine sulfate are added to plasma, the appearance of fibrin strands or gelation (paracoagulation) was specific for fibrin monomers and early fibrin degradation products. The test was sensitive to 1 mg/100 ml fibrin monomers and less than 1 mg/100 ml fibrin degradation products. The test utilizes much lower concentrations of protamine sulfate than previously used, which allows fibrin monomers and early fibrin degradation products to polymerize. Fibrinogen and fibrinogen degradation products did not influence the test. The test was considered positive only when fibrin strands appeared,... This content is PDF only. To continue reading please click on the PDF icon. Author, Article, and Disclosure InformationAffiliations: Boston, Mass. PreviousarticleNextarticle Advertisement FiguresReferencesRelatedDetails Metrics 1 May 1971Volume 74, Issue 5Page: 826-826KeywordsAttentionBlood plasmaFibrinFibrinogenProtamines Issue Published: 1 May 1971 PDF DownloadLoading ...