scholarly journals Crotalocytin: recognition and purification of a timber rattlesnake platelet aggregating protein

Blood ◽  
1980 ◽  
Vol 56 (6) ◽  
pp. 1013-1019 ◽  
Author(s):  
AH Schmaier ◽  
W Claypool ◽  
RW Colman
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Clinical Presentation ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Coagulation Factors ◽  
Single Chain ◽  
Intravascular Coagulation ◽  
Timber Rattlesnake ◽  
Activated Platelets ◽  
Low Ionic Strength

Abstract After being envenomated by the timber rattlesnake, a patient was found to have a platelet count of 5000 per microliter, prothrombin time and activated partial thromboplastin time both greater than 150 sec, plasma fibrinogen 0 mg/dl, and fibrinogen split products 2560 microgram/ml. However, this patient did not appear to have acute disseminated intravascular coagulation since coagulation factors II-XII were normal. We postulated that this venom contained, in addition to a fibrinogen clotting enzyme, a platelet activating protein, Crotalocytin. Crotalocytin was purified from crude timber rattlesnake venom by Sephadex G-100 gel-filtration, low ionic strength precipitation, and DEAE-A50 Sephadex chromatography. By sodium dodecyl sulfate gel electrophoresis and gel-filtration Crotalocytin was a single chain polypeptide, molecular weight 55,000. Thrombocytopenia after timber rattlesnake bite appeared to be due to a protein that directly activated platelets. Timber rattlesnake bite mimicked the clinical presentation of disseminated intravascular coagulation.

scholarly journals Crotalocytin: recognition and purification of a timber rattlesnake platelet aggregating protein

Blood ◽  
1980 ◽  
Vol 56 (6) ◽  
pp. 1013-1019
Author(s):  
AH Schmaier ◽  
W Claypool ◽  
RW Colman
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Clinical Presentation ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Coagulation Factors ◽  
Single Chain ◽  
Intravascular Coagulation ◽  
Timber Rattlesnake ◽  
Activated Platelets ◽  
Low Ionic Strength

After being envenomated by the timber rattlesnake, a patient was found to have a platelet count of 5000 per microliter, prothrombin time and activated partial thromboplastin time both greater than 150 sec, plasma fibrinogen 0 mg/dl, and fibrinogen split products 2560 microgram/ml. However, this patient did not appear to have acute disseminated intravascular coagulation since coagulation factors II-XII were normal. We postulated that this venom contained, in addition to a fibrinogen clotting enzyme, a platelet activating protein, Crotalocytin. Crotalocytin was purified from crude timber rattlesnake venom by Sephadex G-100 gel-filtration, low ionic strength precipitation, and DEAE-A50 Sephadex chromatography. By sodium dodecyl sulfate gel electrophoresis and gel-filtration Crotalocytin was a single chain polypeptide, molecular weight 55,000. Thrombocytopenia after timber rattlesnake bite appeared to be due to a protein that directly activated platelets. Timber rattlesnake bite mimicked the clinical presentation of disseminated intravascular coagulation.

scholarly journals Simvastatin improves the prognosis of endotoxin-induced disseminated intravascular coagulation by regulating the intestinal microenvironment

2020 ◽  
Author(s):  
Min Xu ◽  
Lili Luo ◽  
Mengyi Du ◽  
Lu Tang ◽  
Jie Zhou ◽  
...  
Keyword(s):  
Bacterial Translocation ◽  
Disseminated Intravascular Coagulation ◽  
Intestinal Barrier ◽  
Plasminogen Activator Inhibitor ◽  
Coagulation Factors ◽  
Organ Damage ◽  
Multiple Organ ◽  
Plasminogen Activator Inhibitor 1 ◽  
Intravascular Coagulation ◽  
Coagulation Dysfunction

Abstract Background: Disseminated intravascular coagulation (DIC) is characterized by extensive endothelial injury and coagulation activation that is primarily caused by infection and can be aggravated by the gut due to increased permeability and bacterial translocation. Studies have shown that statins play an important role in reducing inflammation, protecting the endothelium and improving coagulation. In addition, statins regulate tight junction (TJ) proteins and gut microbes. Therefore, we aimed to investigate whether simvastatin improves DIC prognosis by regulating the intestinal microenvironment. Methods: Mice were administered 20 mg/kg simvastatin by gavage for 2 weeks and then intraperitoneally injected with 50 mg/kg endotoxin. Twelve hours later, cytokine release, coagulation dysfunction, multiple organ damage and survival were assessed. In addition, intestinal barrier and permeability and bacteria and bacteria translocation were evaluated. Results: We found that the severity of endotoxin-induced DIC was significantly improved in simvastatin-pretreated mice, who showed attenuated depletion of coagulation factors and platelets, decreased plasminogen activator inhibitor-1 (PAI-1) expression, reduced organ fibrin deposition and an improved survival rate. In addition, simvastatin reduced epithelial apoptosis, increased TJ gene expression, and upregulated antimicrobial peptides, lysozyme and mucins. Simvastatin-pretreated mice showed increased Lactobacillales counts, while the LPS group had increased numbers of Desulfovibrio and Mucispirillum, which produce harmful toxins and damage the intestinal epithelium and mucosa. Finally, with the decreased intestinal permeability in the simvastatin group, bacterial translocation in the organs and blood was significantly reduced, both in quantity and species. Conclusions: Simvastatin improves DIC prognosis, and the intestinal microenvironment participates in this process.

Measuring tissue factor (factor III) activity in plasma.

1989 ◽  
Vol 35 (9) ◽  
pp. 1897-1900 ◽  
Author(s):  
C Fukuda ◽  
K Iijima ◽  
K Nakamura
Keyword(s):  
Tissue Factor ◽  
Disseminated Intravascular Coagulation ◽  
Coagulation Factors ◽  
High Plasma ◽  
Mean Value ◽  
Chromogenic Substrate ◽  
Intravascular Coagulation ◽  
Endogenous Inhibitors ◽  
Tissue Thromboplastin ◽  
The Mean

Abstract This is a method for measuring tissue factor (TF, Factor III, tissue thromboplastin) activity in plasma by using a chromogenic substrate. As pretreatment, the euglobulin fraction of plasma was prepared by removing endogenous inhibitors and heated at 60 degrees C for 3 min to remove fibrinogen. This allowed us to measure the low TF activity in plasma that could not otherwise be measured. Neither phospholipids nor coagulation factors VII, IX, X, or Xa in the samples interfere. Within-run and day-to-day reproducibility were both good. The mean value obtained by this method for normal persons was 1.02 (SD 0.91) arbitrary units/L. A markedly high plasma TF activity of 20 arb. units/L or more was observed in patients with some types of disseminated intravascular coagulation.

scholarly journals Human platelet osteonectin: release, surface expression, and partial characterization

Blood ◽  
1990 ◽  
Vol 75 (5) ◽  
pp. 1105-1113 ◽  
Author(s):  
RJ Jr Kelm ◽  
KG Mann
Keyword(s):  
Solid Phase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Fluid Phase ◽  
Human Bone ◽  
Bovine Bone ◽  
Least Squares Regression ◽  
Single Chain ◽  
Platelet Surface ◽  
Activated Platelets

Abstract Our laboratory has previously shown that osteonectin, an abundant noncollagenous bone protein, is contained in and secreted from human platelets. In this study, the distribution of osteonectin both in the supernatant and on the platelet surface after activation was measured by fluid-phase and solid-phase radioimmunoassay, respectively. Total cellular osteonectin was determined by RIA of guanidinium chloride extracted platelets and ranged from 0.65 to 2.2 micrograms/10(8) platelets or 135,000 to 457,000 molecules/platelet. Platelets treated with varying concentrations of collagen and thrombin released osteonectin in a dose-dependent fashion. Approximately 61% of the total platelet osteonectin was secreted at saturating concentrations of collagen and thrombin. A small fraction of platelet osteonectin is expressed on the surface of platelets in an activation-specific manner as evidenced by the specific and saturable binding of [125I]-anti- osteonectin monoclonal antibody, IIIA3A8, to thrombin-activated platelets. Based on a non-linear least squares regression analysis of the antibody binding, 2,200 IIIA3A8 molecules, or 0.8% of the total platelet osteonectin, is expressed on the platelet surface on activation. Platelet osteonectin was purified from the supernatant of thrombin-activated platelets by immunoaffinity chromatography. Western blotting of proteins secreted by washed, thrombin-stimulated platelets with IIIA3A8 indicated that the osteonectin molecule released from the platelet is a single chain polypeptide. Comparison of immunopurified platelet osteonectin with isolated bovine bone osteonectin and isolated human bone osteonectin by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that platelet osteonectin has a greater apparent molecular weight than bone osteonectin. The NH2-terminal sequence of immunopurified platelet osteonectin was obtained by automated Edman degradation and is identical to the sequence of human bone osteonectin derived from the cDNA of SaOS-2 cells. Collectively, these data suggest that platelet osteonectin is structurally distinct from bone osteonectin in a region of the molecule at a distance from the NH2-terminus.

Induction of Disseminated Intravascular Coagulation in the Factor XII-deficient Fowl

1973 ◽  
Vol 30 (01) ◽  
pp. 025-035 ◽  
Author(s):  
Fredrik Skjørten ◽  
Stein A. Evensen
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Plasma Proteins ◽  
Coagulation Factors ◽  
Factor Xii ◽  
Contact Activation ◽  
Intravascular Coagulation ◽  
Fibrillar Material ◽  
Ultrastructural Appearance ◽  
Tissue Thromboplastin ◽  
Homologous Tissue

SummaryBirds are naturally deficient in the coagulation factors responsible for the contact activation reactions in mammalian plasma. In the present study, fowl lungs were examined for evidence of disseminated intravascular coagulation (DIC) 5 min or 4 hours after injection of either Liquoid or bacterial endotoxin. These substances are potent initiators of DIC in mammals, and activation of factor XII is believed to be essential for their triggering effect.Liquoid injection produced intravascular deposits with the light microscopical staining properties of fibrin. However these deposits had a purely granular ultra-structure; their formation was not prevented by adequate anticoagulation, and there was no concomitant thrombocyte aggregation. It is suggested that the deposits represent precipitates of plasma proteins, including fibrinogen.Endotoxin failed to produce clinical reactions, intravascular deposits or thrombocyte aggregates. In contrast, animals injected with homologous tissue thromboplastin died, and fibrillar material with the ultrastructural appearance of fibrin, as well as thrombocyte aggregates were found in small pulmonary vessels. These effects were completely prevented by anticoagulation.We conclude that both Liquoid and endotoxin failed to trigger DIC in the factor XII-deficient fowl, suggesting that these substances depend on the contact activation reactions for the generation of thrombin.

Studies on the Purification and Characterization of Human Urinary Plasminogen and Plasmin

1979 ◽  
Vol 42 (05) ◽  
pp. 1536-1547
Author(s):  
Hsin Fu Chen ◽  
Masao Nakabayashi ◽  
Kazuo Satoh ◽  
Shoichi Sakamoto
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Gel Electrophoresis ◽  
Specific Activity ◽  
Gel Filtration ◽  
New Method ◽  
Chronic Glomerulonephritis ◽  
Purification And Characterization ◽  
Intravascular Coagulation ◽  
Human Plasminogen

SummaryA new method is described for the preparation of highly purified human plasminogen and plasmin with specific activity of 32 CTA units per mg of protein. With this method, the purification of the urinary plasminogen + plasmin antigenic materials from patients with chronic glomerulonephritis, disseminated intravascular coagulation syndrome and severe toxemia of pregnancy was performed, and the resulting highly purified proenzyme and enzyme were analyzed by immunoelectrophoresis, separative agar electrophoresis, gel filtration and SDS-gel electrophoresis.Our findings indicated that urinary plasmin reflects more closely the extent of intraglomerular fibrinolysis, while urinary plasminogen reflects non-selective proteinuria in patients with chronic glomerulonephritis or severe toxemia of pregnancy.

Kinetics Of Antithrombin III During Severe Consumption Coagulopathy In An Infant Measured Without Radioactive Tracer Proteins

1981 ◽  
Author(s):  
B Schmidt ◽  
U Wais ◽  
I Witt ◽  
W Pringsheim ◽  
W Künz
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Antithrombin Iii ◽  
Radioactive Tracer ◽  
Coagulation Factors ◽  
Turnover Rates ◽  
Intravascular Coagulation ◽  
Elimination Half ◽  
Coagulation Proteins ◽  
Precise Diagnosis ◽  
Severe Coagulopathy

The precise diagnosis of disseminated intravascular coagulation remains difficult to achieve: Decreased production of coagulation factors and/or thrombocytes can mimic the laboratory pattern of disseminated intravascular coagulation. The measurement of increased turnover rates for coagulation proteins would provide more convincing criteria.During the course of severe coagulopathy in an infant suffering from septicaemia and shock, antithrombin levels were determined repeatedly before and during treatment with Antithrombin concentrate: Activities and concentrations were measured, using chromogenic substrates and immunodiffusion plates, respectively. By mathematical analysis of these data, using a biexponential function, the plasma elimination half-life of the antithrombin III was estimated to be 7.5 to 10.5 hours. Compared with known plasma half-lives of radioactively labelled antithrombin III in adults, the increase was five- to ten-fold. This indicates an accelerated consumption of antithrombin III in this case of severe coagulopathy.

Disseminated Intravascular Coagulation in Cancer: An Update

2019 ◽  
Vol 45 (04) ◽  
pp. 342-347 ◽  
Author(s):  
Marcel Levi
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Inflammatory Responses ◽  
Traumatic Injury ◽  
Distant Metastases ◽  
Coagulation Factors ◽  
Bleeding Complications ◽  
Adequate Treatment ◽  
Intravascular Coagulation ◽  
Coagulation Proteins ◽  
Thrombotic Complications

AbstractCancer often leads to the activation of coagulation, manifesting as disseminated intravascular coagulation (DIC) in its most extreme form. DIC is characterized by systemic intravascular coagulation activation (leading to deposition of intravascular platelets and fibrin) and simultaneous consumption of coagulation proteins and thrombocytes (which may cause bleeding complications). The clinical course of DIC in patients with malignancies is typically less intense compared with DIC complicating alternative clinical settings, including systemic inflammatory responses following infection or traumatic injury. A more slowly proceeding, less fulminant, and widespread hemostatic derangement can remain asymptomatic. Eventually, the ongoing consumption may result in low levels of platelets and coagulation factors, and bleeding complications (frequently localized at the site of the tumor or distant metastases) may be the first clinical manifestation of DIC. An alternative clinical scenario is dominated by thrombotic complications, ranging from clinically manifest vascular thrombosis to microvascular platelet plugs. The main principle of DIC management is adequate treatment of the precipitating disorder; however, there are clinical presentations that may require additional supportive strategies specifically aimed at the amelioration of the coagulopathy.

scholarly journals Subunit composition and structure of subcomponent C1q of the first component of human complement

1976 ◽  
Vol 155 (1) ◽  
pp. 19-23 ◽  
Author(s):  
K B M Reid ◽  
R R Porter
Keyword(s):  
Ionic Strength ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Helical Structure ◽  
Guanidinium Chloride ◽  
Molecular Weights ◽  
Composition And Structure ◽  
Covalently Linked ◽  
Low Ionic Strength ◽  
Expected Position

1. Unreduced human subcomponent C1q was shown by electrophoresis on polyacrylamide gels run in the presence of sodium dodecyl sulphate to be composed of two types of non-covalently linked subunits of apparent mol.wts. 69 000 and 54 000. The ratio of the two subunits was markedly affected by the ionic strength of the applied sample. At a low ionic strength of applied sample, which gave the optimum value for the 54 000-apparent mol.wt. subunit, a ratio of 1.99:1.00 was obtained for the ratio of the 69 000-apparent mol.wt. subunit to the 5400-apparent-mol.wt. subunit. The amount of the 54 000-apparent-mol.wt. subunit detected in the expected position on the gel was found to be inversely proportional to increases in the ionic strength of the applled sample. 2. Human subcomponent C1q on reduction and alkylation, or oxidation, yields equimolar amounts of three chains designated A, B and C [Reid et al. (1972) Biochem. J. 130, 749-763]. The results obtained by Yonemasu & Stroud [(1972) Immunochemistry 9, 545-554], which showed that the 69 000-apparent-mol.wt. subunit was a disulphide-linked dimer of the A and B chains and that the 54 000-apparent-mol.wt. subunit was a disulphide-linked dimer of the C chain, were confirmed. 3. Gel filtration on Sephadex G-200 in 6.0M-guanidinium chloride showed that both types of unreduced subunit were eluted together as a single symmetrical peak of apparent mol.wt. 49 000-50 000 when globular proteins were used as markers. The molecular weights of the oxidized or reduced A, B and C chains have been shown previously to be very similar all being in the range 23 000-24 000 [Reid et al. (1972) Biochem. J. 130, 749-763; Reid (1974) Biochem. J. 141, 189-203]. 4. It is proposed that subcomponent C1q (mol.wt. 410000) is composed of nine non-covalently linked subunits, i.e. six A-B dimers and three C-C dimers. 5. A structure for subcomponent C1q is proposed and is based on the assumption that the collagen-like regions of 78 residues in each of the A, B and C chains are combined to form a triple-helical structure of the same type as is found in collagens.

The Elimination of Labelled Fibrinogen in Galactosamine-Induced Experimental Hepatitis in Rabbits

1975 ◽  
Author(s):  
I. Mahn ◽  
C. Reuter ◽  
H. Merkel ◽  
G. Müller-Berghaus
Keyword(s):  
Liver Disease ◽  
Intravenous Injection ◽  
Disseminated Intravascular Coagulation ◽  
Isotonic Saline ◽  
Coagulation Factors ◽  
Virus Hepatitis ◽  
Intravascular Coagulation ◽  
Coagulation Defect ◽  
Deutsche Forschungsgemeinschaft ◽  
Glomerular Capillaries

The intravenous injection of D-galactosamine-HCl into animals induces a liver disease resembling virus hepatitis in man in its histological and clinico-phatological features. In a previous study disseminated intravascular coagulation was demonstrated by tracing fibrin-rich microdots in the renal glomerular capillaries, especially if the fibrinolytic system was inhibited by EACA (Thromb. Res. 1, 473, 1972). In order to differentiate between disturbance of synthesis and disseminated intravascular coagulation, investigations with 125I-fibrinogen were performed in rabbits treated with D-galactosamine (1 g/kg) and EACA (0.5 g/kg xhr). In rabbits infused with galactosamine and EACA the elimination of 125I-fibrinogen was increased in comparison to the control animals treated with EACA or isotonic saline only. If heparin (750 u/kg × hr) was infused additionally to galactosamine and EACA, the accelerated decay of labelled fibrinogen was prevented. The occurrence of 125I-activity in organs was pronounced in animals exhibiting microdot formation. These experiments indicate that due to a diminished synthesis of coagulation factors in this model of hepatitis disseminated intravascular coagulation may contribute to the coagulation defect.(Supported by the Deutsche Forschungsgemeinschaft, Bad Godesberg, Germany.)

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