Protamine Sulfate Test Research Articles

Some features of the hemostasis system in pregnant women at risk of developing preeclampsia

The objective: a study of the hemostasis system in pregnant women with a risk of preeclampsia development.Materials and methods. 100 pregnant women with the risk for preeclampsia (main group) were examined. The risk factors were determined according to the Guideline “Hypertensive Disorders During Pregnancy”, Order No. 676 of the Ministry of Health of Ukraine. The control group contained of 50 healthy women with physiological pregnancy. The groups were representative in age and reproductive history.The following indicators of hemostasis were studied: the platelet system (the number of platelets, their aggregation ability and the total platelet aggregation index (TPAI), the coagulation system (autocoagulation test, thrombin time, prothrombin index, fibrinogen concentration) and the state of the fibrinolysis system which was determined by such indicators: plasma level of free heparin, activity of antithrombin III, indicators of ethanol and protamine sulfate tests, concentration of soluble fibrin in blood plasma.Results. In pregnant women with a risk of preeclampsia, there are changes in platelet hemostasis indicators: a significant decrease in the number of platelets and a significant (p<0.05) increase in platelet aggregation ability, there is a tendency to an increase in TPAI indicators. In the main group a significant increase in the fibrinogen concentration, plasma lysis indicators and a tendency to an increase of the free heparin concentration, a decrease of antithrombin III and, in comparison with the indicators in healthy women, a 3-fold increase in the content of soluble fibrin (p<0.05) were found.Conclusions. In pregnant women with a risk of preeclampsia development, there are disorders in the vascular-platelet hemostasis, coagulation and fibrinolytic blood systems, namely, a significant tension in the platelet link of the system, an increase in thrombogenic potential, and a sharp inhibition of the fibrinolytic link of hemostasis.

Postmortem diagnosis of shock conditions

Aim. Develop a method for postmortem diagnosis of shock conditions.
 Methods. Brood serum obtained from 72 dead bodies of people who died from different causes such as anaphylaxis, toxic shock, cardiovascular diseases, traumatic injury and thermal burns was examined. The level of fibrinogen fraction was performed by sulfite-alkaline method. Plasma soluble fibrin monomer complexes were identified by paracoagulation tests (ethanol and protamine sulfate tests).
 Results. The method of shock conditions postmortem diagnosis was determined, regardless of shock etiology. The lowest levels of fibrinogen fraction in cases of anaphylactic and toxic shock were 19.3 and 11.9 g/l, respectively, which is significantly higher compared to other groups. In cases of death due to acute burn, fibrinogen level was almost tenfold higher (p

Evaluation of point-of-care tests for diagnosis of disseminated intravascular coagulation in dogs admitted to an intensive care unit

Objective To evaluate the accuracy of point-of-care tests for the diagnosis of disseminated intravascular coagulation (DIC) in dogs and assess the correlation and agreement of results between point-of-care and laboratory tests in the evaluation of hemostatic function. Design Prospective case series. Animals 59 critically ill dogs (affected dogs) with clinical signs of diseases known to predispose to DIC and 52 clinically normal dogs. Procedures Accuracy of the point-of-care tests (activated clotting time [ACT], estimated platelet count and number of schizocytes from a blood smear, plasma total solids [TS] concentration, and the protamine sulfate test) was evaluated, using receiver operating characteristic curves and likelihood ratios. A strategy, using likelihood ratios to calculate a post-test probability of DIC, was tested with 65% used as a threshold for initiation of treatment. Results of laboratory tests (coagulogram and plasma antithrombin III activity) were used as the standard for comparison in each dog. Results ACT and estimated platelet count provided the best accuracy for detection of DIC. The plasma TS concentration, schizocyte number, and protamine sulfate test had poor accuracy. The strategy using post-test probability of DIC identified 12 of 16 affected dogs that had DIC. Estimated platelet count was correlated and had acceptable clinical agreement with automated platelet count (r = 0.70). The plasma TS (r = 0.28) concentration and serum albumin (r = 0.63) concentration were not accurate predictors of plasma antithrombin III activity. The ACT did not correlate with activated partial thromboplastin time (r = 0.28). Conclusions and Clinical Relevance Strategic use of likelihood ratios from point-of-care tests can assist clinicians in making treatment decisions for dogs suspected to have DIC when immediate laboratory support is unavailable. (J Am Vet Med Assoc 1999;215:805–810)

Diagnostic efficacy of the D-Dimer assay in disseminated intravascular coagulation (DIC)

The D-Dimer (D-D) assay for measuring cross-linked fibrin degradation products is now available for the clinical laboratory. We combined this assay with other tests to assess patients with diagnosed or suspected DIC. Also, a small group of patients (20) with deep venous thrombosis (DVT) were studied. The D-D test, antithrombin-III assay, FDP titer, fibrinopeptide-A level, protamine sulfate test, fibrinogen, prothrombin time, and activated partial thromboplastin time were used. The D-D test was abnormal in 93.7%, the AT-III level was abnormal in 87.5%, the fibrinopeptide-A level was abnormal in 89.5%, and the FDP titer was elevated in 83.7% of patients with DIC. When assessing patients found not to have confirmed DIC the D-D assay was abnormal in 20%, the AT-III level was abnormal in 6%, and the fibrinopeptide-A level was elevated in 13%. We conclude the D-Dimer assay to be a useful molecular marker of hemostasis in diagnosing DIC and this test will often discriminate between those patients with or without DIC, especially when used with the AT-III and fibrinopeptide-A assays. Of the battery of tests used in this study, the most useful, in descending order of efficacy, appear to be the D-dimer assay (93.7% abnormal), the fibrinopeptide-A titer (89.5% abnormal), the AT-III level (87.5% abnormal), and the FDP titer (83.7% abnormal).

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.

Postpartum hemolytic uremic syndrome

A 22-year-old. gravida 1, para 1, abortus 1 woman was readmitted to the hospital with abdominal pain, vomiting, and bloody diarrhea of 2 days' duration. Pregnancy had been uncomplicated; blood pressure readings and urinalyses showed normal results. Labor and delivery at 39 weeks' gestation had been uncomplicated, and the patient had been discharged on the third postpartum day. Examination at readmission on the fifth postpartum day revealed diffuse abdominal tenderness with rebound and hypoactive bowel sounds. The patient was afebrile, and results of a test for occult blood in the stool were positive. Laboratory data included hematocrit, 48%; platelet count, 67,000/mm3 ; white blood cell count, 21 ,300/mm3 ; creatinine, 1.0 mg/dl; normal electrolyte count; urinalysis results showing protein uria (4+), 13 to 20 white blood cells per high-power field, 15 to 20 red blood cells per high-power field, and hyaline and granular casts. A coagulation profile revealed a normal fibrinogen level (308 mg/dl), a normal prothrombin time (12.5 seconds), normal protamine sulfate test results, and elevated fibrin split products (10 to 40 ng/ml). AsCites was present on x-ray evaluation of the abdomen. Cultures of urine, blood, stool, and ascitic fluid showed no growth. During the first 5 days after admission the hematocrit de-

Comparative study of paracoagulation tests in acute myocardial infarction

Ethanol, -naphthol (test for "fibrinogen B") and protamine sulfate tests, performed simultaneously in the blood plasma of patients with acute myocardial infarction, showed a high degree of correlation with each other, but they differed in sensitivity: ethanol -naphthol protamine sulfate. Heparin did not distort the results of the ethanol and -naphthol tests, while the protamine sulfate test results strongly depended on the presence of heparin. The formation of a granular precipitate is not a positive protamine sulfate test and indicates hyperheparinemia. The ethanol test on the basis of data can be recommended to monitor the state of the hemocoagulation system in acute myocardial infarction, including the period of heparin therapy.

The influence of disseminated intravascular coagulation on renal function after experimental hemorrhagic shock.

Abstract Experiments were performed to investigate the influence of disseminated intravascular coagulation (DIC) during shock upon renal function after shock. Hemorrhagic hypotension of 2 h duration at 50 mmHg was induced in anesthetized non-heparinized dogs, followed by reinfusion of shed blood (shock group 1). In shock group 2, acute splenectomy was performed prior to hemorrhage and epsilon-amino caproic acid was infused during hemorrhagic hypotension. In shock group 1, during hemorrhagic hypotension the whole blood clotting time and thrombin time shortened (hypercoagulability), the arterial platelet count increased and intrarenal platelet retention was observed. The results of renal function measurements reproduced the well known pattern of renal dysfunction after hemorrhagic shock in dogs: reduction of CPAH to about 55% and of CI to about 70% of pre-shock levels, significant increases of VU, ENa, E F Na (diuresis and natriuresis), decreases of U P Osm and TH2Oc and increase of V U C I and C Osm C I (disturbance of concentrating mechanism). Dogs in shock group 2 developed prolongation of whole blood and plasma clotting times (hypocoagulability), decrease in platelet count and factors V and VIII activity (factor depletion), and increases in titers of FDP (staphylococcal clumping test assay) and fibrin monomers (protamine sulfate test assay). Clotting changes were consistent with the diagnosis of DIC. After reinfusion, a different renal dysfunction pattern was observed: CPAH and CI were both reduced to about 35% of their pre-shock levels and CI as well as EI and EPAH were significantly lower than in shock group 1 and the increase of urine output did not occur. The depression of renal function and urine excretion following shock with DIC could be explained by pronounced cortical ischemia and reproduced the characteristic features of the acute renal failure in man (depression of EPAH, low CI, reduction of C I TRPF , decrease of U P Osm and TH2Oc). It is concluded, that coagulation derangements in addition to vasoconstriction during shock determine the course of renal function after shock and play an important role in the pathogenesis of shock kidney.

Evaluation of paracoagulation tests by plasma fibrinogen chromatography

Performance of ethanol gelation test (EG test) and of a diluted protamine sulfate test (DPS test), was assessed by quantitative plasma fibrinogen chromatography by testing a large series of unrelated plasma samples. The relationship between paracoagulation tests and chromatographic findings was highly significant (p < 0.001). However both sensitivity and specificity of EG and DPS tests were shown to depend on fibrinogen concentration; EG test appeared less sensitive at low fibrinogen levels whereas DPS test appeared less sensitive at high fibrinogen levels.

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.

Clinico-experimental study on prevention of the complications following incompatible blood transfusion

The accident of incompatible blood transfusion is not infrequent in practical medicine. The acute hemolytic transfusion reaction sometimes results severe complications. In authors' experience in the treatment and clinical analysis of this disorder during last one decade, there were some evidences that disseminated intravascular coagulation attributed in considerable extent to the development of such complications as shock, hemorrhagic tendencies and acute renal failure. In this paper, six patients received incompatible blood transfusion are reviewed and some therapeutic approaches are presented.On the hypothesis that DIC is a primary pathogenetic mechanism involved in these hemolytic reaction, authors have learned that the heparin therapy would be the best at interfering with these pathophysiologic pathways, and have been getting the benifits in prevention of severe complications or of exaggeration of signs and symptoms developed. Of 6 cases presented, five of the incompatibilities were in ABO system and one in Rh system. A 26-year-old female, blood group 0, Rh positive, was given a transfusion with 0 positive blood but she was positive in anti-E antibody. The amount of blood transfused were 50-350ml in ABO incompatibilities and 1, 000ml of E-positive blood in a Rh-incompatible case.Hemostatic studies were done in 3 patients. One patient received ABO incompatible blood revealed clotting abnormalities even 2 days after the heparin administration. The case with positive E-antibody developed acute renal failure. Severe bleeding tendency was observed at the Cesarian section when she was given more than 1, 500ml of blood. Thrombocytopenia, positive tests in FDP and protamine sulfate test and prolongation of thrombin time and serial thrombin time were seen in these 2 cases. Remaining one patient who was given heparin just after the accident showed no coagulation disturbance when tested during heparin therapy. Five patients received mismatched blood in ABO system were all given heparin as soon as possible after the accident to prevent severe reactions due to DIC. All were symptomatically uneventful, however, the elevation of FDP and thrombocytopenia were noted in one as mentioned above and moderate increase of blood urea nitrogen in three clinical courses. These findings seem to represent the subclinical participation of clotting episodes in mismatched transfusion reactions even after the heparin administration. Therefore, the occurrence of DIC may be more frequent that has been appreciated and prompt treatment with heparin prior to the recognition of signs and symptoms may decrease the morbidity and mortality from incompatible transfusion reactions.

Hemostatic Abnormalities in Malignancy, a Prospective Study of One Hundred Eight Patients: Part I. Coagulation Studies

A prospective study of hemostatic abnormalities in 108 cancer patients was undertken at an oncology clinic in a university teaching hospital. Tests included Quick prothrombin time, activated partial thromboplastin time, thrombin time, platelet count, modified Ivy bleeding time, fibrinogen, fibrin degradation products (FDP), euglobulin lysis time, protamine sulfate test, and factor V, VII, VIII and X assays. Ninety-eight per cent of the patients had one or more abnormal coagulation tests. The commonest abnormalities were elevated fibrin degradation products and prolonged thrombin time. Thrombocytosis occurred in 57% of patients, hyperfibrinogenemia in 46%, thrombocytopenia in 11%, and non had hypofibrinogenmia. It is suggested that platelet count, fibrinogen concentration, and serum FDP assay are the most useful tests in assessing the hemostatic abnormalities in cancer patients, although thrombin time, factor V assay, and bleeding time may also be helpful. The peripheral blood smears of 53 patients were reviewed, and only one showed microangiopathic hemolytic anemia. The data illustrate that subclinical coagulopathy is relatively frequent in patients with malignancy.

Uterine and Peripheral Hematologic Profiles in Molar Pregnancy

The potential differences in hematologic profiles of blood samples drawn simultaneously from the right utero-ovarian vein and from the upper extremity were investigated in four patients with uncomplicated molar pregnancy in stable obstetric conditions. The patients had undergone no previous chemotherapy and were scheduled for total abdominal hysterectomies. The dominant abnormalities in uterine venous blood were prolongation of thrombin time; shortening of activated partial thromboplastin time; positive protamine sulfate test; and increase in coagulation factors II and VII, with a tendency to low values in factor V. Peripheral samples gave almost parallel results in all altered and normal tests, except in one case with very striking differences in factors II, V, VII and X. Several local and systemic influences are discussed. It is concluded that molar pregnancy seems to have important systemic mechanisms affecting the stability of the blood coagulation homeostasis, which act in addition to those at a local level.

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.

Coagulation and fibrinolysis in hepatic cirrhosis

In order to clarify coagulation and fibrinolysis in hepatic cirrhosis following items were examined.1) Counts of megakaryocyte and its classification. 2) Lifespan of 5Cr-labeled platelet. 3) Platelet volume and its distribution (Coulter Counter ZB). 4) Platelet spreading test (Breddin's method). 5) Platelet aggregation (Evans Aggregometer). 6) Platelet Factor-3 (Kaolin). 7) Serum phospholipids and its fractionation. 8) Hepaplastin test (Eisai pharmaceutical Co, test for II, Vll, X factors) 9) Plasma fibrinogen (Tyrosine). 10) FDP (HIT). 11) Protamine sulfate test (Lipinski's). 12) Total plasmin (fibrin plate). 13) Plasminogen and antiplasmin (lysine sepharose affinity chromatography) 14) Factor XIII (Urayama's). from these examinations following facts were clarified.1) Reduction of platelet counts is due to disturbance of platelet formation and enhanced breakdown of platelets and may either due to pooling in the spleen.The remarkable reduction and its characteristic asthenic pattern of platelet aggregation may suggest hypofunction of the platelet cell. It may be supported with the facts of the large spreading tendency of the platelet and reduction of PF-3 availability.2) Hepaplastin test clearly shows reduction of extrinsic factor in cirrhosis of the liver.3) In regard to f ibrinolysis, in case of the hepatic cirrhosis, plasma fibrinogen tends to decrease, on the other hand protamind sulfate test tend to show higher value.Total plasma plasmin increased remarkably, on the other hand, plasminogen and antiplasmin slightly decreased. Factor XIII decreased either in case of cirrhosis.Whether the enhanced fibrinolysis may occur primary or secondary must be examined more in detail.

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.

Abnormal coagulation and fibrinolysis in eclampsia: A clinical and laboratory correlation study

This study evaluated the existence of abnormally increased coagulation and fibrinolysis in 33 severely toxemic and eclamptic women by means of a combined hemotologic profile with clinical and morphologic correlations. The dominant findings were: different degrees of thrombocytopenia, abnormal levels of blood fibrinogen, prolonged thrombin time, and positive protamine sulfate test. Altered activated partial thromboplastin time and positive ethanol gelation test were slightly less frequent, and only few cases showed prolonged prothrombin time or early lysis of euglobulins. These abnormalities seemed to be more numerous and more pronounced in the worst cases of the series and their severity seemed to be associated with the age of the patient and the presence of previous underlying disease. These variously handicapped pregnant women exhibited worse hematologic hematologic abnormalities, and provided most of the fatal cases in the series. Finally, the main findings were discussed and commented upon.

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.