158 results on '"Mann, Kenneth G."'
Search Results
2. Thrombosis: theoretical considerations
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Mann, Kenneth G.
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Thrombosis -- Health aspects ,Hemostasis -- Health aspects ,Von Willebrand factor -- Physiological aspects ,Food/cooking/nutrition ,Health - Abstract
The blood coagulation process is initiated in response to vascular injury and results in either hemostasis or thrombosis. The process can be divided conceptually into separate steps including initiation, propagation, termination, elimination, and repair. Concise descriptions of each of these processes are provided in the present review together with an attempt to integrate these processes at a conceptual level so as to avoid the unfortunate tendency to apply linear logic to the complex temporal interplay among the various processes in the blood coagulation system. Am J Clin Nutr 1997;65(suppl):1657S-64S.
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- 1997
3. Hemorrhagic events during therapy with recombinant tissue-type plasminogen activator, heparin, and aspirin for acute myocardial infarction: results of the Thrombolysis in Myocardial Infarction (TIMI), Phase II Trial
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Bovill, Edwin G., Terrin, Michael L., Stump, David C., Berke, Andrew D., Frederick, Margaret, Collen, Desire, Feit, Frederick, Gore, Joel M., Hillis, L. David, Lambrew, Costas T., Leiboff, Roy, Mann, Kenneth G., Markis, John E., Pratt, Craig M., Sharkey, Scott W., Sopko, George, Tracy, Russell P., and Chesebro, James H.
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Hemorrhage -- Risk factors ,Thrombolytic therapy -- Complications ,Heart attack -- Drug therapy ,Tissue plasminogen activator -- Adverse and side effects ,Health - Abstract
In recent years, several large scale studies have demonstrated marked improvement in survival of patients with heart attacks who are treated with medications that dissolve the blood clots blocking the coronary arteries. One of these drugs is recombinant tissue-type plasminogen activator, or rt-PA, and it was used in the Thrombolysis in Myocardial Infarction, Phase II trial (TIMI II). Drugs like rt-PA not only dissolve existing clots, they prevent new clots from forming; consequently, patients given such drugs are at increased risk of bleeding complications. The rate of bleeding complications in the patients who participated in TIMI II was studied. The earliest participants in the study were given doses of 150 milligrams (mg) of rt-PA, while the later participants received 100 mg of rt-PA. Some patients had cardiac catheterization tests early in the course of their recovery, while others underwent these tests later, if at all (the TIMI II trial showed that there was no benefit to having the test done early). Bleeding complications were seen in approximately four percent of the TIMI II patients, regardless of the rt-PA dose. However, those who had early cardiac catheterization experienced the most frequent and most severe bleeding problems. Other factors associated with higher risk of bleeding complications were female gender, weight less than 70 kilograms (154 pounds), decompensating heart function and low platelet count (platelets are small blood cells necessary for successful blood clotting). A prior history of high blood pressure was associated with a greater risk of bleeding complications. Most of the bleeding complications occurred within three days of treatment. Among the serious bleeding complications noted were stroke, cardiac tamponade (in which blood collects in the sac surrounding the heart, preventing it from pumping effectively), and gastrointestinal bleeding. Some patients had laboratory evidence of blood loss, with decreases in their blood counts, but had no identifiable source of bleeding. To minimize the risk of bleeding, further modification of rt-PA treatment is warranted. (Consumer Summary produced by Reliance Medical Information, Inc.)
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- 1991
4. Relationship of glucocorticoid dosage to serum bone GLA-protein concentration in patients with rheumatologic disorders
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Kotowicz, Mark A., Hall, Stephen, Hunder, Gene G., Cedel, Sandra L., Mann, Kenneth G., and Riggs, B. Lawrence
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Prednisone -- Adverse and side effects ,Corticosteroids -- Adverse and side effects ,Bones -- Growth ,Osteoporosis -- Causes of ,Health - Abstract
Osteoporosis (reduction of bone mass) is a serious complication of treatment with glucocorticoid (steroid) drugs. Bone loss tends to occur in the ribs and vertebrae, and may lead to fractures at these sites. One study showed that vertebral or rib fractures occurred in 45 percent of asthmatic patients treated with glucocorticoids. Osteopenia (decreased bone tissue) was detected in 38 percent of patients treated with steroids. Glucocorticoid treatment may interfere with the absorption and elimination of calcium, resulting in hyperparathyroidism (increased activity of the parathyroid gland, which controls bone metabolism). Hyperparathyroidism may result in increased bone resorption and decreased bone formation, leading to bone loss. Other possible mechanisms underlying bone loss include increased sensitivity to the actions of parathyroid hormone or direct effects of glucocorticoids on the osteoblasts, cells involved in bone formation. Bone formation can be assessed by measuring blood levels of bone GLA-protein, also known as osteocalcin (BGP), a bone protein produced by osteoblasts. Previous studies have shown decreased blood levels of BGP in patients treated with glucocorticoids. The relationship between blood levels of BGP and steroid drug dosage was assessed to determine the extent to which bone formation was suppressed for each dose. A statistical model was developed to estimate the extent of blood BGP level suppression when steroids were administered. Using this model, a suppression of 50 percent was predicted with dosages of 20 to 25 milligrams per day of the steroid drug prednisone. (Consumer Summary produced by Reliance Medical Information, Inc.)
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- 1990
5. Thrombin *: can't live without it; probably die from it
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Mann, Kenneth G.
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Blood clot -- Physiological aspects -- Usage ,Anticoagulants (Medicine) -- Physiological aspects -- Usage ,Thrombosis -- Physiological aspects -- Usage ,Warfarin -- Usage -- Physiological aspects ,Thrombin -- Physiological aspects -- Usage ,Health ,Usage ,Physiological aspects - Abstract
Thrombin is a multifaceted protein with a wide range of functions. Walter Seegers, a pioneer in thrombin work, referred to it as 'the living enzyme of my blood.' (1) The [...]
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- 2003
6. Endogenous Procoagulant Activity in Trauma Patients and Its Relationship to Trauma Severity
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Prior, Shannon M., Park, Myung S., Mann, Kenneth G., and Butenas, Saulius
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- 2019
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7. Microparticles formed during storage of red blood cell units support thrombin generation
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Bouchard, Beth A., Orfeo, Thomas, Keith, Hollis N., Lavoie, Elizabeth M., Gissel, Matthew, Fung, Mark, and Mann, Kenneth G.
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Supplemental digital content is available in the text.
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- 2018
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8. Probing the Dynamics of Clot-Bound Thrombin at Venous Shear Rates
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Haynes, Laura M., Orfeo, Thomas, Mann, Kenneth G., Everse, Stephen J., and Brummel-Ziedins, Kathleen E.
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In closed system models of fibrin formation, exosite-mediated thrombin binding to fibrin contributes to clot stability and is resistant to inhibition by antithrombin/heparin while still susceptible to small, active-site inhibitors. Each molecule of fibrin can bind ∼1.6 thrombin molecules at low-affinity binding sites (Kd = 2.8 μM) and ∼0.3 molecules of thrombin at high-affinity binding sites (Kd = 0.15 μM). The goal of this study is to assess the stability of fibrin-bound thrombin under venous flow conditions and to determine both its accessibility and susceptibility to inhibition. A parallel-plate flow chamber (7 × 50 × 0.25 mm) for studying the stability of thrombin (0–1400 nM) adhered to a fibrin matrix (0.1–0.4 mg/mL fibrinogen, 10 nM thrombin) under a variety of venous flow conditions was developed using the thrombin-specific, fluorogenic substrate SN-59 (100 μM). The flow within this system is laminar (Re < 1) and reaction rates are driven by enzyme kinetics (Pe = 100, Da = 7000). A subpopulation of active thrombin remains stably adhered to a fibrin matrix over a range of venous shear rates (46–184 s−1) for upwards of 30 min, and this population is saturable at loads >500 nM and sensitive to the initial fibrinogen concentration. These observations were also supported by a mathematical model of thrombin adhesion to fibrin, which demonstrates that thrombin initially binds to the low-affinity thrombin binding sites before preferentially equilibrating to higher affinity sites. Antithrombin (2.6 μM) plus heparin (4 U/mL) inhibits 72% of the active clot-bound thrombin after ∼10 min at 92 s−1, while no inhibition is observed in the absence of heparin. Dabigatran (20 and 200 nM) inhibits (50 and 93%) clot-bound thrombin reversibly (87 and 66% recovery). This model illustrates that clot-bound thrombin stability is the result of a constant rearrangement of thrombin molecules within a dense matrix of binding sites.
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- 2017
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9. What are the abnormal thrombotic responses in the clotting system of individuals with atherosclerosis? Are these responses genetically determined or adaptations to the disease state?
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Mann, Kenneth G.
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Thrombosis -- Genetic aspects ,Blood clotting -- Genetic aspects ,Atherosclerosis -- Genetic aspects ,Food/cooking/nutrition ,Health - Published
- 1992
10. TACTIC: Trans‐Agency Consortium for Trauma‐Induced Coagulopathy
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Mann, K.G., Freeman, K., Mann, Kenneth G., Esmon, Charles T., Wisnewski, Stephen, Tracy, Russell P., Kindzelski, Andrei L., Pusateri, Anthony, Banerjee, Anirban, Brass, Lawrence F., Brummel‐Ziedins, Kathleen E., Butenas, Saulius, Cohen, Mitchell J., Diamond, Scott L., Freeman, Kalev, Moore, Ernest E., Morrissey, James H., Nelson, Mark T., Park, Myung S., Ruf, Wolfram, Shupp, Jeffrey W., Sperry, Jason L., Spiess, Bruce D., Stalker, Timothy J., and Zuckerbraun, Brian S.
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Trauma‐induced coagulopathy (TIC) includes heterogeneous coagulopathic syndromes with different underlying causes, and treatment is challenged by limited diagnostic tests to discriminate between these entities in the acute setting. We provide an overview of progress in understanding the mechanisms of TIC and the context for several of the hypotheses that will be tested in ‘TACTIC’. Although connected to ongoing clinical trials in trauma, TACTIC itself has no intent to conduct clinical trials. We do anticipate that ‘early translation’ of promising results will occur. Functions anticipated at this early translational level include: (i) basic science groundwork for future therapeutic candidates; (ii) development of acute coagulopathy scoring systems; (iii) coagulation factor composition‐based computational analysis; (iv) characterization of novel analytes including tissue factor, polyphosphates, histones, meizothrombin and α‐thrombin–antithrombin complexes, factor XIa, platelet and endothelial markers of activation, signatures of protein C activation and fibrinolysis markers; and (v) assessment of viscoelastic tests and new point‐of‐care methods.
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- 2015
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11. Anticoagulant effects of statins and their clinical implications
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Undas, Anetta, Brummel-Ziedins, Kathleen E., and Mann, Kenneth G.
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- 2014
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12. Anti-FVIII antibodies in Black and White hemophilia A subjects: Do F8haplotypes play a role?
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Pratt, Kathleen P., Gunasekera, Devi, Vir, Pooja, Tan, Siyuan, Pierce, Glenn F., Olsen, Cara, Butenas, Saulius, and Mann, Kenneth G.
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The most common complication in hemophilia A (HA) treatment, affecting 25-30% of severe HA patients, is the development of alloimmune inhibitors that foreclose the ability of infused factor VIII (FVIII) to participate in coagulation. Inhibitors confer significant pathology on affected individuals and present major complexities in their management. Inhibitors are more common in African American patients, and it has been hypothesized that this is a consequence of haplotype (H)-treatment product mismatch. F8haplotypes H1-H5 are defined by nonsynonymous single-nucleotide polymorphisms encoding sequence variations at FVIII residues 1241, 2238 and 484. Haplotypes H2-H5 are more prevalent in individuals with black African ancestry, while 80-90% of the white population has the H1 haplotype. This study used an established multiplex fluorescence immunoassay to determine anti-FVIII antibody titers in plasma from 394 HA subjects (188 black, 206 white), measuring their binding to recombinant full-length H1 and H2 and B-domain-deleted (BDD) H1/H2, H3/H5 and H4 FVIII proteins. Inhibitor titers were determined using a chromogenic assay and linear B-cell epitopes characterized using peptide microarrays. FVIII-reactive antibodies were readily detected in most HA subjects, with higher titers in those with a current inhibitor, as expected. Neither total nor inhibitory antibody titers correlated with F8haplotype mismatches, and peptides with D1241E and M2238V polymorphisms did not comprise linear B-cell epitopes. Interestingly, the BDD-FVIII proteins were markedly more reactive than the full-length FVIII products with plasma antibodies. The stronger immunoreactivity of BDD-FVIII suggests that B-domain removal may expose novel B-cell epitopes, perhaps through conformational rearrangements of FVIII domains.
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- 2022
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13. Prothrombin activation in blood coagulation: the erythrocyte contribution to thrombin generation
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Whelihan, Matthew F., Zachary, Vicentios, Orfeo, Thomas, and Mann, Kenneth G.
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Prothrombin activation can proceed through the intermediates meizothrombin or prethrombin-2. To assess the contributions that these 2 intermediates make to prothrombin activation in tissue factor (Tf)–activated blood, immunoassays were developed that measure the meizothrombin antithrombin (mTAT) and α-thrombin antithrombin (αTAT) complexes. We determined that Tf-activated blood produced both αTAT and mTAT. The presence of mTAT suggested that nonplatelet surfaces were contributing to approximately 35% of prothrombin activation. Corn trypsin inhibitor–treated blood was fractionated to yield red blood cells (RBCs), platelet-rich plasma (PRP), platelet-poor plasma (PPP), and buffy coat. Compared with blood, PRP reconstituted with PPP to a physiologic platelet concentration showed a 2-fold prolongation in the initiation phase and a marked decrease in the rate and extent of αTAT formation. Only the addition of RBCs to PRP was capable of normalizing αTAT generation. FACS on glycophorin A–positive cells showed that approximately 0.6% of the RBC population expresses phosphatidylserine and binds prothrombinase (FITC Xa·factor Va). These data indicate that RBCs participate in thrombin generation in Tf-activated blood, producing a membrane that supports prothrombin activation through the meizothrombin pathway.
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- 2012
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14. Guided Antithrombotic Therapy
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Fuster, Valentin, Bhatt, Deepak L., Califf, Robert M., Michelson, Alan D., Sabatine, Marc S., Angiolillo, Dominick J., Bates, Eric R., Cohen, David J., Coller, Barry S., Furie, Bruce, Hulot, Jean-Sebastien, Mann, Kenneth G., Mega, Jessica L., Musunuru, Kiran, O'Donnell, Christopher J., Price, Matthew J., Schneider, David J., Simon, Daniel I., Weitz, Jeffrey I., Williams, Marlene S., Hoots, W. Keith, Rosenberg, Yves D., and Hasan, Ahmed A.K.
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- 2012
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15. Clotting Factor Deficiency in Early Trauma-Associated Coagulopathy
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Rizoli, Sandro B., Scarpelini, Sandro, Callum, Jeannie, Nascimento, Bartolomeu, Mann, Kenneth G., Pinto, Ruxandra, Jansen, Jan, and Tien, Homer C.
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Coagulopathic bleeding is a leading cause of in-hospital death after injury. A recently proposed transfusion strategy calls for early and aggressive frozen plasma transfusion to bleeding trauma patients, thus addressing trauma-associated coagulopathy (TAC) by transfusing clotting factors (CFs). This strategy may dramatically improve survival of bleeding trauma patients. However, other studies suggest that early TAC occurs by protein C activation and is independent of CF deficiency. This study investigated whether CF deficiency is associated with early TAC.
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- 2011
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16. Dilutional Control of Prothrombin Activation at Physiologically Relevant Shear Rates
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Haynes, Laura M., Dubief, Yves C., Orfeo, Thomas, and Mann, Kenneth G.
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The generation of proteolyzed prothrombin species by preassembled prothrombinase in phospholipid-coated glass capillaries was studied at physiologic shear rates (100–1000 s−1). The concentration of active thrombin species (α-thrombin and meizothrombin) reaches a steady state, which varies inversely with shear rate. When corrected for shear rate, steady-state levels of active thrombin species exhibit no variation and a Michaelis-Menten analysis reveals that chemistry of this reaction is invariant between open and closed systems; collectively, these data imply that variations with shear rate arise from dilutional effects. Significantly, the major products observed include nonreactive species arising from the loss of prothrombin's phospholipid binding domain (des F1 species). A numerical model developed to investigate the spatial and temporal distribution of active thrombin species within the capillary reasonably approximates the observed output of total thrombin species at different shears; it also predicts concentrations of active thrombin species in the wall region sufficient to account for observed levels of des FI species. The predominant feedback formation of nonreactive species and high levels of the primarily anticoagulant intermediate meizothrombin (∼40% of total active thrombin species) may provide a mechanism to prevent thrombus propagation downstream of a site of thrombosis or hemorrhage.
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- 2011
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17. Cellular regulation of blood coagulation: a model for venous stasis
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Campbell, James E., Brummel-Ziedins, Kathleen E., Butenas, Saulius, and Mann, Kenneth G.
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We have adapted the corn-trypsin inhibitor whole-blood model to include EA.hy926 as an endothelium surrogate to evaluate the vascular modulation of blood coagulation initiated by relipidated recombinant tissue factor (rTf) and a cellular Tf surrogate, lipopolysaccharide (LPS)-stimulated THP1 cells (LPS-THP-1). Compared with bare tubes, EA.hy926 with rTf decreased the rate of thrombin formation, ITS accumulation, and the production of fibrinopeptide A. These phenomena occurred with increased rates of factor Va (fVa) inactivation by cleavages at R506 and R306. Thus, EA.hy926 provides thrombin-dependent protein C activation and APC fVa inactivation. Comparisons of rTf with LPS-THP-1 showed that the latter gave reduced rates for TAT formation but equivalent fibrinopeptide A, and fV activation/inactivation. In the presence of EA.hy926, the reverse was obtained; with the surrogate endothelium and LPS-THP-1 the rates of TAT generation, fibrinopeptide release, and fV activation were almost doubled, whereas cleavage at R306 was equivalent. These observations suggest cooperativity between the 2 cell surrogates. These data suggest that the use of these 2 cell lines provides a reproducible quasi-endothelial quasi-inflammatory cytokine-stimulated monocyte system that provides a method to evaluate the variations in blood phenotype against the background of stable inflammatory cell activator and a stable vascular endothelial surrogate.
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- 2010
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18. Thrombin generation in rheumatoid arthritis: Dependence on plasma factor composition
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Undas, Anetta, Gissel, Matthew, Kwasny-Krochin, Beata, Gluszko, Piotr, Mann, Kenneth G., and Brummel-Ziedins, Kathleen E.
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- 2010
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19. Thromboelastography as a Better Indicator of Hypercoagulable State After Injury Than Prothrombin Time or Activated Partial Thromboplastin Time
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Park, Myung S., Martini, Wenjun Z., Dubick, Michael A., Salinas, Jose, Butenas, Saulius, Kheirabadi, Bijan S., Pusateri, Anthony E., Vos, Jeffrey A., Guymon, Charles H., Wolf, Steven E., Mann, Kenneth G., and Holcomb, John B.
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To investigate the hemostatic status of critically ill, nonbleeding trauma patients. We hypothesized that a hypercoagulable state exists in patients early after severe injury and that the pattern of clotting and fibrinolysis are similar between burned and nonburn trauma patients.
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- 2009
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20. Quantitation of anti–factor VIII antibodies in human plasma
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Krudysz-Amblo, Jolanta, Parhami-Seren, Behnaz, Butenas, Saulius, Brummel-Ziedins, Kathleen E., Gomperts, Edward D., Rivard, Georges E., and Mann, Kenneth G.
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The presence of antibodies (Abs) in hemophilia A patients can potentially influence the therapeutic qualities of factor VIII (fVIII) administration. Much work has been focused on the presence of inhibitory antibodies, whereas the quantitation of noninhibitory anti-fVIII antibodies has been largely undetermined. Our objective was to develop a sensitive and specific fluorescence-based immunoassay (FLI) for the quantitation of anti-fVIIIAbs in human plasma. Affinity-purified human anti-fVIIIAb, isolated from a hemophilia A subject, was used as a calibrator with a detectability limit of 40 (±1.5) pM. The calibrator and the human plasma anti-fVIIIAb were captured on recombinant fVIII (rfVIII)– coupled microspheres and probed with mouse anti–human Ig–R-phycoerythrin. Plasma samples from 150 healthy donors and 39 inhibitor-negative hemophilia A subjects were compared with 4 inhibitor-positive hemophilia A plasma samples with inhibitor titers of 1 BU/mL (94.6 ± 0.8 nM), 11 BU/mL (214.3 ± 7.1 nM), 106 BU/mL (2209.4 ± 84.9 nM), 140 BU/mL (2417.7 ± 3.8 nM) as measured by the Nijmegen method. We also describe the validation of a mouse anti–human fVIIIAb as a surrogate calibrator. Four healthy individuals (3%) showed detectable anti-fVIIIAb in the range of 0.6 to 6.2 nM, whereas 13 (33%) of the 39 inhibitor-free hemophilia A subjects were positive for anti-fVIIIAb in the range of 0.5 to 20 nM. The method may be useful for therapeutic management of hemophilia A patients.
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- 2009
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21. Systemic blood coagulation activation in acute coronary syndromes
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Undas, Anetta, Szułdrzyński, Konstanty, Brummel-Ziedins, Kathleen E., Tracz, Wiesława, Zmudka, Krzysztof, and Mann, Kenneth G.
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We evaluated systemic alterations to the blood coagulation system that occur during a coronary thrombotic event. Peripheral blood coagulation in patients with acute coronary thrombosis was compared with that in people with stable coronary artery disease (CAD). Blood coagulation and platelet activation at the microvascular injury site were assessed using immunochemistry in 28 non-anticoagulated patients with acute myocardial infarction (AMI) versus 28 stable CAD patients matched for age, sex, risk factors, and medications. AMI was associated with increased maximum rates of thrombin-antithrombin complex generation (by 93.8%; P < .001), thrombin B-chain formation (by 57.1%; P < .001), prothrombin consumption (by 27.9%; P = .012), fibrinogen consumption (by 27.0%; P = .02), factor (f) Va light chain generation (by 44.2%; P = .003), and accelerated fVa inactivation (by 76.1%; P < .001), and with enhanced release of platelet-derived soluble CD40 ligand (by 44.4%; P < .001). FVa heavy chain availability was similar in both groups because of enhanced formation and activated protein C (APC)–mediated destruction. The velocity of coagulant reactions in AMI patients showed positive correlations with interleukin-6. Heparin treatment led to dampening of coagulant reactions with profiles similar to those for stable CAD. AMI-induced systemic activation of blood coagulation markedly modifies the pattern of coagulant reactions at the site of injury in peripheral vessels compared with that in stable CAD patients.
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- 2009
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22. Factor XIa and tissue factor activity in patients with coronary artery disease
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Butenas, Saulius, Undas, Anetta, Gissel, Matthew T, Szuldrzynski, Konstanty, Zmudka, Krzysztof, and Mann, Kenneth G
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- 2008
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23. Antithrombotic properties of aspirin and resistance to aspirin: beyond strictly antiplatelet actions
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Undas, Anetta, Brummel-Ziedins, Kathleen E., and Mann, Kenneth G.
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Aspirin is effective in the prevention of cardiovascular events in high-risk patients. The primary established effect of aspirin on hemostasis is to impair platelet aggregation via inhibition of platelet thromboxane A2 synthesis, thus reducing thrombus formation on the surface of the damaged arterial wall. Growing evidence also indicates that aspirin exerts additional antithrombotic effects, which appear to some extent unrelated to platelet thromboxane A2 production. Aspirin can reduce thrombin generation with the subsequent attenuation of thrombin-mediated coagulant reactions such as factor XIII activation. Aspirin also acetylates lysine residues in fibrinogen resulting in increased fibrin clot permeability and enhanced clot lysis as well as directly promoting fibrinolysis with high-dose aspirin. The variable effectiveness of aspirin in terms of clinical outcomes and laboratory findings, which has been termed aspirin resistance, may be related to these additional antithrombotic effects that are altered when associated with common genetic polymorphisms such as the Leu33Pro β3-integrin or Val34Leu factor XIII mutations. However, the clinical relevance of these observations is still unclear. Elucidation of the actual impacts of aspirin other than antiaggregation effects could be important in view of the widespread use of this drug in the prevention of thrombotic manifestations of atherosclerosis.
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- 2007
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24. Blood coagulation at the site of microvascular injury in healthy and coumadin-treated subjects heterogenous for factor V Leiden mutation
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Undas, Anetta, Brzezinska-Kolarz, Beata, Orfeo, Tom, Brummel-Ziedins, Kathleen, Musial, Jacek, Szczeklik, Andrew, and Mann, Kenneth G.
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- 2007
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25. Identification of an inactivating cleavage site for α-thrombin on the heavy chain of factor Va
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Erdogan, Evrim, Bukys, Michael A., Orfeo, Thomas, Mann, Kenneth G., and Kalafatis, Michael
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- 2007
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26. The Resuscitative Fluid You Choose May Potentiate Bleeding
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Brummel-Ziedins, Kathleen, Whelihan, Matthew F., Ziedins, Eduards G., and Mann, Kenneth G.
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Trauma is the leading cause of death in the younger population in the United States, frequently from the development of hemorrhagic shock. Controversy exists over the type of volume resuscitation for restoring hemodynamic stability that should be used in hemorrhagic shock. Little is known about how various resuscitative paradigms affect the coagulation cascade, which is essential to controlling hemorrhagic shock.
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- 2006
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27. Tissue factor activity in whole blood
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Butenas, Saulius, Bouchard, Beth A., Brummel-Ziedins, Kathleen E., Parhami-Seren, Behnaz, and Mann, Kenneth G.
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Tissue factor (TF) is an integral membrane protein essential for hemostasis. During the past several years, a number of studies have suggested that physiologically active TF circulates in blood at concentrations greater than 30 pM either as a component of blood cells and microparticles or as a soluble plasma protein. In our studies using contact pathway–inhibited blood or plasma containing activated platelets, typically no clot is observed for 20 minutes in the absence of exogenous TF. An inhibitory anti-TF antibody also has no effect on the clotting time in the absence of exogenous TF. The addition of TF to whole blood at a concentration as low as 16 to 20 fM results in pronounced acceleration of clot formation. The presence of potential platelet TF activity was evaluated using ionophore-treated platelets and employing functional and immunoassays. No detectable TF activity or antigen was observed on quiescent or ionophore-stimulated platelets. Similarly, no TF antigen was detected on mononuclear cells in nonstimulated whole blood, whereas in lipopolysaccharide (LPS)–stimulated blood a significant fraction of monocytes express TF. Our data indicate that the concentration of physiologically active TF in non–cytokine-stimulated blood from healthy individuals cannot exceed and is probably lower than 20 fM.
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- 2005
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28. A review of the therapeutic uses of thrombin
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Lundblad, Roger L., Bradshaw, Ralph A., Gabriel, Don, Ortel, Thomas L., Lawson, Jeffrey, and Mann, Kenneth G.
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- 2004
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29. Factor V: a combination of Dr Jekyll and Mr Hyde
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Mann, Kenneth G. and Kalafatis, Michael
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- 2003
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30. Thrombin functions during tissue factor–induced blood coagulation
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Brummel, Kathleen E., Paradis, Sara G., Butenas, Saulius, and Mann, Kenneth G.
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Tissue factor–induced blood coagulation was studied in 20 individuals, for varying periods of time during 54 months, in contact pathway–inhibited whole blood at 37°C and evaluated in terms of the activation of various substrates. After quenching over time with inhibitors, the soluble phases were analyzed for thrombin–antithrombin III (TAT) complex formation, prothrombin fragments, platelet activation (osteonectin release), factor Va generation, fibrinopeptide (FP) A and FPB release, and factor XIII activation. TAT complex formation, for 35 experiments, showed an initiation phase (up to 4.6 ± 0.6 minutes) in which thrombin was generated at an average rate of 0.93 ± 0.3 nM/min catalyzed by about 1.3 pM prothrombinase yielding approximately 26 nM thrombin. During a subsequent propagation phase, thrombin was generated at a rate of 83.9 ± 3.8 nM/min by about 120 pM prothrombinase, reaching ultimate levels of 851 ± 53 nM. Clot time, determined subjectively, occurred at 4.7 ± 0.2 minutes and correlated with the inception of the propagation phase. The thrombin concentrations associated with the transitions to rapid product formation are 510 ± 180 pM for platelet activation (1.9 ± 0.2 minutes), 840 ± 280 pM for factor XIII activation and factor Va generation (2.2 ± 0.6 minutes), 1.3 ± 0.4 nM for FPA release (2.5 ± 0.7 minutes), 1.7 ± 0.5 nM for FPB release and prethrombin 2 (2.8 ± 0.8 minutes), 7.0 ± 2.2 nM for thrombin B chain (3.6 ± 0.2 minutes), and 26 ± 6.2 nM for the propagation phase of TAT formation (4.6 ± 0.6 minutes). These results illustrate that the initial activation of thrombin substrates occurs during the initiation phase at less than 2 nM thrombin (0.2%). Most thrombin (96%) is formed well after clotting occurs.
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- 2002
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31. A Model for the Stoichiometric Regulation of Blood Coagulation*
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Hockin, Matthew F., Jones, Kenneth C., Everse, Stephen J., and Mann, Kenneth G.
- Abstract
We have developed a model of the extrinsic blood coagulation system that includes the stoichiometric anticoagulants. The model accounts for the formation, expression, and propagation of the vitamin K-dependent procoagulant complexes and extends our previous model by including: (a) the tissue factor pathway inhibitor (TFPI)-mediated inactivation of tissue factor (TF)·VIIa and its product complexes; (b) the antithrombin-III (AT-III)-mediated inactivation of IIa, mIIa, factor VIIa, factor IXa, and factor Xa; (c) the initial activation of factor V and factor VIII by thrombin generated by factor Xa-membrane; (d) factor VIIIa dissociation/activity loss; (e) the binding competition and kinetic activation steps that exist between TF and factors VII and VIIa; and (f) the activation of factor VII by IIa, factor Xa, and factor IXa. These additions to our earlier model generate a model consisting of 34 differential equations with 42 rate constants that together describe the 27 independent equilibrium expressions, which describe the fates of 34 species. Simulations are initiated by “exposing” picomolar concentrations of TF to an electronic milieu consisting of factors II, IX, X, VII, VIIa, V, and VIIII, and the anticoagulants TFPI and AT-III at concentrations found in normal plasma or associated with coagulation pathology. The reaction followed in terms of thrombin generation, proceeds through phases that can be operationally defined as initiation, propagation, and termination. The generation of thrombin displays a nonlinear dependence upon TF, AT-III, and TFPI and the combination of these latter inhibitors displays kinetic thresholds. At subthreshold TF, thrombin production/expression is suppressed by the combination of TFPI and AT-III; for concentrations above the TF threshold, the bolus of thrombin produced is quantitatively equivalent. A comparison of the model with empirical laboratory data illustrates that most experimentally observable parameters are captured, and the pathology that results in enhanced or deficient thrombin generation is accurately described.
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- 2002
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32. Mechanism of factor VIIa–dependent coagulation in hemophilia blood
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Butenas, Saulius, Brummel, Kathleen E., Branda, Richard F., Paradis, Sara G., and Mann, Kenneth G.
- Abstract
The ability of factor VIIa to initiate thrombin generation and clot formation in blood from healthy donors, blood from patients with hemophilia A, and in anti–factor IX antibody–induced (“acquired”) hemophilia B blood was investigated. In normal blood, both factor VIIa–tissue factor (TF) complex and factor VIIa alone initiated thrombin generation. The efficiency of factor VIIa was about 0.0001 that of the factor VIIa–TF complex. In congenital hemophilia A blood and “acquired” hemophilia B blood in vitro, addition of 10 to 50 nM factor VIIa (pharmacologic concentrations) corrected the clotting time at all TF concentrations tested (0-100 pM) but had little effect on thrombin generation. Fibrinopeptide release and insoluble clot formation were only marginally influenced by addition of factor VIIa. TF alone had a more pronounced effect on thrombin generation; an increase in TF from 0 to 100 pM increased the maximum thrombin level in “acquired” hemophilia B blood from 120 to 480 nM. Platelet activation was considerably enhanced by addition of factor VIIa to both hemophilia A blood and “acquired” hemophilia B blood. Thus, pharmacologic concentrations of factor VIIa cannot restore normal thrombin generation in hemophilia A and hemophilia B blood in vitro. The efficacy of factor VIIa (10-50 nM) in hemophilia blood is dependent on TF.
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- 2002
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33. Blood coagulation at the site of microvascular injury: effects of low-dose aspirin
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Undas, Anetta, Brummel, Kathleen, Musial, Jacek, Mann, Kenneth G., and Szczeklik, Andrew
- Abstract
The sequence of coagulant reactions in vivo following vascular injury is poorly characterized. Using quantitative immunoassays, the time courses were evaluated for activation of prothrombin, factor (F)V, FXIII, fibrinogen (Fbg) cleavage, and FVa inactivation in bleeding-time blood collected at 30-second intervals from 12 healthy subjects both before and after aspirin ingestion. Prothrombin decreased at a maximum rate of 14.2 ± 0.6 nM per second to 10% of initial values at the end of bleeding. Significant amounts of α-thrombin B chain appeared rapidly at 90 seconds of bleeding and increased at a maximum rate of 0.224 ± 0.03 nM per second to a peak value of 38 nM. Kinetics of prethrombin 2 generation was almost identical. Prothrombinase concentration reached a peak value of 22 pM at 150 seconds and then decreased to 9 pM at the end of bleeding. Prothrombin fragment 1.2 (F1.2) was produced explosively (0.673 ± 0.05 nM per second), whereas thrombin-antithrombin III (TAT) complexes were generated at a much slower rate (0.11 ± 0.008 nM per second;P= .002). FVa light chain was detectable 30 seconds later than the heavy chain (150 seconds) and was produced at a slightly slower rate (0.027 ± 0.001 nM per second) when compared with the heavy chain (0.032 ± 0.002 nM per second; P= .041). The 30 000 fragment (residues 307-506) of FVa heavy chain produced by activated protein C appeared as early as at 90 seconds and increased with time. Fbg was removed from the blood shed with a high rate of 0.047 ± 0.02 μM/s and became undetectable at approximately 180 seconds of bleeding. The velocity of FXIII activation correlated with thrombin B-chain formation. A 7-day aspirin administration (75 mg/d) resulted in significant reductions in maximum rates of (1) prothrombin removal (by 29%; P= .008); generation of α-thrombin B-chain (by 27.2%; P= .022), and prethrombin 2 (by 26%; P= .014); formation of F1.2 (by 31.4%;P= .009) and TAT (by 30.3%; P= 0.013); (2) release of FVa heavy chain (by 25%; P= .003) and FVa light chain (by 29.6%; P= .007); (3) Fbg depletion from solution (by 30.5%; P= .002); and (4) FXIII activation (by 28.6%; P= .003). Total amounts of the proteins studied, collected at every interval, also significantly decreased following aspirin ingestion. These results indicate that low-dose aspirin impairs thrombin generation and reactions catalyzed by this enzyme at the site of the injury.
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- 2001
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34. The Role of the Membrane in the Inactivation of Factor Va by Plasmin
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Kalafatis, Michael and Mann, Kenneth G.
- Abstract
The mechanism of inactivation of bovine factor Va by plasmin was studied in the presence and absence of phospholipid vesicles (PCPS vesicles). Following 60-min incubation with plasmin (4 nm) membrane-bound factor Va (400 nm) is completely inactive, whereas in the absence of phospholipid vesicles following a 1-h incubation period, the cofactor retains 90% of its initial cofactor activity. Amino acid sequencing of the fragments deriving from cleavage of factor Va by plasmin demonstrated that while both chains of factor Va are cleaved by plasmin, only cleavage of the heavy chain correlates with inactivation of the cofactor. In the presence of a membrane surface the heavy chain of the bovine cofactor is first cleaved at Arg348to generate a fragment of Mr47,000 containing the NH2-terminal part of the cofactor (amino acid residues 1–348) and a Mr42,000 fragment (amino acid residues 349–713). This cleavage is associated with minimal loss in cofactor activity. Complete loss of activity of the membrane-bound cofactor coincides with three cleavages at the COOH-terminal portion of the Mr47,000 fragment: Lys309, Lys310, and Arg313. These cleavages result in the release of the COOH terminus of the molecule and the production of a Mr40,000 fragment containing the NH2-terminal portion of the factor Va molecule. Factor Va was treated with plasmin in the absence of phospholipid vesicles followed by the addition of PCPS vesicles and activated protein C (APC). A rapid inactivation of the cofactor was observed as a result of cleavage of the Mr47,000 fragment at Arg306by APC and appearance of aMr39,000 fragment. These data suggest a critical role of the amino acid sequence 307–348 of factor Va. A 42-amino acid peptide encompassing the region 307–348 of human factor Va (N42R) was found to be a good inhibitor of factor Va clotting activity with an IC50of ∼1.3 μm. These data suggest that plasmin is a potent inactivator of factor Va and that region 307–348 of the cofactor plays a critical role in cofactor function and may be responsible for the interaction of the cofactor with factor Xa and/or prothrombin.
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- 2001
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35. Antiplatelet agents in tissue factor–induced blood coagulation
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Butenas, Saulius, Cawthern, Kevin M., van't Veer, Cornelis, DiLorenzo, Maria E., Lock, Jennifer B., and Mann, Kenneth G.
- Abstract
Several platelet inhibitors were examined in a tissue factor (TF)–initiated model of whole blood coagulation. In vitro coagulation of human blood from normal donors was initiated by 25 pM TF while contact pathway coagulation was suppressed using corn trypsin inhibitor. Products of the reaction were analyzed by immunoassay. Preactivation of platelets with the thrombin receptor activation peptide did not influence significantly the clotting time or thrombin–antithrombin III complex (TAT) formation. Addition of prostaglandin E1 (5 μM) caused a significant delay in clotting (10.0 minutes) versus control (4.3 minutes). The prolonged clotting time is correlated with delays in platelet activation, formation of TAT, and fibrinopeptide A (FPA) release. In blood from subjects receiving acetylsalicylic acid (ASA or aspirin), none of the measured products of coagulation were significantly affected. Similarly, no significant effect was observed when 5 μM dipyridamole (Persantine) was added to the blood. Antagonists of the platelet integrin receptor glycoprotein (gp) IIb/IIIa had intermediate effects on the reaction. A 1- to 2-minute delay in clot time and FPA formation was observed with addition of the antibodies 7E3 and Reopro (abciximab) (10 μg/mL), accompanied by a 40% to 70% reduction in the maximal rate of TAT formation and delay in platelet activation. The cyclic heptapetide, Integrilin (eptifibatide), at 5 μM concentration slightly prolonged clot time and significantly attenuated the maximum rate of TAT formation. The disruption of the gpIIb/IIIa-ligand interaction not only affects platelet aggregation, but also decreases the rate of TF-initiated thrombin generation in whole blood, demonstrating a potent antithrombotic effect superimposed on the antiaggregation characteristics.
- Published
- 2001
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36. Homocysteine Inhibits Inactivation of Factor Va by Activated Protein C*
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Undas, Anetta, Williams, E. Brady, Butenas, Saulius, Orfeo, Thomas, and Mann, Kenneth G.
- Abstract
We report the effect of homocysteine on the inactivation of factor Va by activated protein C (APC) using clotting assays, immunoblotting, and radiolabeling experiments. Homocysteine, cysteine, or homocysteine thiolactone have no effect on factor V activation by α-thrombin. Factor Va derived from homocysteine-treated factor V was inactivated by APC at a reduced rate. The inactivation impairment increased with increasing homocysteine concentration (pseudo first order rate k= 1.2, 0.9, 0.7, 0.4 min−1at 0, 0.03, 0.1, 1 mmhomocysteine, respectively). Neither cysteine nor homocysteine thiolactone treatment of factor V affected APC inactivation of derived factor Va. Western blot analyses of APC inactivation of homocysteine-modified factor Va are consistent with the results of clotting assays. Factor Va, derived from factor V treated with 1 mmβ-mercaptoethanol was inactivated more rapidly than the untreated protein sample. Factor V incubated with [35S]homocysteine (10–450 μm) incorporated label within 5 min, which was found only in those fragments that contained free sulfhydryl groups: the light chain (Cys-1960, Cys-2113), the B region (Cys-1085), and the 26/28-kDa (residues 507–709) APC cleavage products of the heavy chain (Cys-539, Cys-585). Treatment with β-mercaptoethanol removed all radiolabel. Plasma of patients assessed to be hyperhomocysteinemic showed APC resistance in a clot-based assay. Our results indicate that homocysteine rapidly incorporates into factor V and that the prothrombotic tendency in hyperhomocysteinemia may be related to impaired inactivation of factor Va by APC due to homocysteinylation of the cofactor by modification of free cysteine(s).
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- 2001
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37. Platelets and Phospholipids in Tissue Factor-initiated Thrombin Generation
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Butenas, Saulius, Branda, Richard F, van ’t Veer, Cornelis, Cawthern, Kevin M, and Mann, Kenneth G
- Published
- 2001
- Full Text
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38. The Regulation of the Factor VII-Dependent Coagulation Pathway: Rationale for the Effectiveness of Recombinant Factor VIIa in Refractory Bleeding Disorders
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Van 't Veer, Cornelis and Mann, Kenneth G.
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- 2000
- Full Text
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39. Inhibition of thrombin generation by the zymogen factor VII: implications for the treatment of hemophilia A by factor VIIa
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van 't Veer, Cornelis, Golden, Neal J., and Mann, Kenneth G.
- Abstract
Factor VII circulates as a single chain inactive zymogen (10 nmol/L) and a trace (∼10-100 pmol/L) circulates as the 2-chain form, factor VIIa. Factor VII and factor VIIa were studied in a coagulation model using plasma concentrations of purified coagulation factors with reactions initiated with relipidated tissue factor (TF). Factor VII (10 nmol/L) extended the lag phase of thrombin generation initiated by 100 pmol/L factor VIIa and low TF. With the coagulation inhibitors TFPI and AT-III present, factor VII both extended the lag phase of the reaction and depressed the rate of thrombin generation. The inhibition of factor Xa generation by factor VII is consistent with its competition with factor VIIa for TF. Thrombin generation with TF concentrations >100 pmol/L was not inhibited by factor VII. At low tissue factor concentrations (<25 pmol/L) thrombin generation becomes sensitive to the absence of factor VIII. In the absence of factor VIII, factor VII significantly inhibits TF-initiated thrombin generation by 100 pmol/L factor VIIa. In this hemophilia A model, approximately 2 nmol/L factor VIIa is needed to overcome the inhibition of physiologic (10 nmol/L) factor VII. At 10 nmol/L, factor VIIa provided a thrombin generation response in the hemophilia model (0% factor VIII, 10 nmol/L factor VII) equivalent to that observed with normal plasma, (100% factor VIII, 10 nmol/L factor VII, 100 pmol/L factor VIIa). These results suggest that the therapeutic efficacy of factor VIIa in the medical treatment of hemophiliacs with inhibitors is, in part, based on overcoming the factor VII inhibitory effect.
- Published
- 2000
- Full Text
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40. Issues with the Assay of Factor VIII Activity in Plasma and Factor VIII Concentrates
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Lundblad, Roger L., Kingdon, Henry S., Mann, Kenneth G., and White, Gilbert C.
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- 2000
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41. Regulation of Prothrombinase Activity by Protein S
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van't Veer, Cornelis, Butenas, Saulius, Golden, Neal J., and Mann, Kenneth G.
- Published
- 1999
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42. “Normal” Thrombin Generation
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Butenas, Saulius, van’t Veer, Cornelis, and Mann, Kenneth G.
- Abstract
We have investigated the influence of alterations in plasma coagulation factor levels between 50% and 150% of their mean values for prothrombin, factor X, factor XI, factor IX, factor VII, factor VIII, factor V, protein C, protein S, antithrombin III (AT-III), and tissue factor pathway inhibitor (TFPI) as well as combinations of extremes, eg, 50% anticoagulants and 150% procoagulants or 50% procoagulants and 150% anticoagulants in a synthetic “plasma” system. The reaction systems were constructed in vitro using purified, natural, and recombinant proteins and synthetic phospholipid vesicles or platelets with the reactions initiated by recombinant tissue factor (TF)-factor VIIa complex (5 pmol/L). To investigate the influence of the protein C system, soluble thrombomodulin (Tm) was also added to the reaction mixture. For the most extreme situations in which the essential plasma procoagulants (prothrombin, and factors X, IX, V, and VIII) and the stoichiometric anticoagulants (AT-III and TFPI) were collectively and inversely altered by 50%, a 28-fold difference in the total available thrombin generated was observed. Variations of most of these proteins 50% above and below the “normal” range, with the remainder at 100%, had only modest influences on the peak and total levels of thrombin generated. The dominant factors influencing thrombin generation were prothrombin and AT-III. When these 2 components were held at 100% and all other plasma procoagulants were reduced to 50%, there was a 60% reduction in the available thrombin generated. No increase in the thrombin generated was observed when the 150% level of all plasma procoagulants other than prothrombin was evaluated. When only prothrombin was raised to 150%, and all other factors were maintained at 100%, the thrombin generated increased by 71% to 121%. When AT-III was at 50% and all other constituents were at 100%, thrombin production was increased by 104% to 196%. The additions of protein C and protein S over the 50% to 150% ranges with Tm at 0.1 nmol/L concentration had limited influence on thrombin generation. Individual variations in factors VII, XI, and X concentrations had little effect on the duration of the initiation phase, the peak thrombin level achieved, or the available thrombin generated. Paradoxically, increases in factor IX concentration to 150% led to lowered thrombin generation, while decreases to 50% led to enhanced thrombin generation, most likely a consequence of factor IX as a competitive substrate with factor X for factor VIIa-TF. Reductions in factor V or factor VIII concentration led to prolongations of the initiation phase, while the reduction of TFPI to 50% led to shortening of this phase. However, none of these alterations led to significant changes in the available thrombin generated. Based on these data, one might surmise that increases in prothrombin and reductions in AT-III, within the normal range, would be potential risk factors for thrombosis and that algorithms that combine normal factor levels may be required to develop predictive tests for thrombosis.
- Published
- 1999
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43. Biochemistry and Physiology of Blood Coagulation
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Mann, Kenneth G.
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- 1999
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44. Monoclonal Antibodies to Human Coagulation Factor V and Factor Va
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Foster, W. Barry, Tucker, Marjorie M., Katzmann, Jerry A., Miller, Randall S., Nesheim, Michael E., and Mann, Kenneth G.
- Abstract
BALB/c mice were immunized with human factor V. The immunogen was a mixture of procofactor (factor V) and thrombin-activated cofactor (factor Va). Spleen cells were obtained from an immunized animal and fused with NS-1 murine myeloma cells. Hybrid cell cultures were assayed for the production of antibodies to human factor V and factor Va by a solid-phase radioimmunoassay. Factor V and/or factor-Va-specific antibodies were detected in 38 of the 96 cultures assayed. The cells from 10 of these positive cultures were subcloned by limiting dilution and grown as ascites tumors in BALB/c mice. Ascitic fluids were obtained and characterized with respect to their binding interaction with human factor V and factor Va. Three hybridoma cell lines produce monoclonal antibodies that react equally well with factor V and factor Va. Another antibody reacts with both antigens, but the reactivity with factor V is better than with factor Va. An additional two antibodies react with factor Va better than factor V in the radioimmunoassay (RIA). The remaining four antibodies react exclusively with factor V. A previously described murine monoclonal antibody to human factor V (αHFV-1) has been used to study the peptides produced during the thrombin-catalyzed activation of human factor V. This antibody binds both factor V and factor Va, releases them at high ionic strength, and has an apparent dissociation constant for factor Va of 3 × 10-9M. When human factor V (mol wt 330,000) is activated by thrombin and passed over an αHFV-1-Sepharose affinity resin, factor Va binds and subsequently can be eluted. The eluate in 1.2 MNaCI contains two fragments of apparent mol wt 93,000 and 70,000. EDTA, which inactivates factor Va, promotes release of the mol wt 93,000 fragment from factor Va bound to the antibody. Subsequent elution with 1.2 MNaCI releases the mol wt 70,000 fragment. These observations indicate that human factor Va is a two subunit protein and that the epitope for αHFV-1 is on the mol wt 70,000 fragment.
- Published
- 1983
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45. Blood Coagulation in Hemophilia A and Hemophilia C
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Cawthern, Kevin M., van ‘t Veer, Cornelis, Lock, Jennifer B., DiLorenzo, Maria E., Branda, Richard F., and Mann, Kenneth G.
- Abstract
Tissue factor (TF)-induced coagulation was compared in contact pathway suppressed human blood from normal, factor VIII-deficient, and factor XI-deficient donors. The progress of the reaction was analyzed in quenched samples by immunoassay and immunoblotting for fibrinopeptide A (FPA), thrombin-antithrombin (TAT), factor V activation, and osteonectin. In hemophilia A blood (factor VIII:C <1%) treated with 25 pmol/L TF, clotting was significantly delayed versus normal, whereas replacement with recombinant factor VIII (1 U/mL) restored the clot time near normal values. Fibrinopeptide A release was slower over the course of the experiment than in normal blood or hemophilic blood with factor VIII replaced, but significant release was observed by the end of the experiment. Factor V activation was significantly impaired, with both the heavy and light chains presenting more slowly than in the normal or replacement cases. Differences in platelet activation (osteonectin release) between normal and factor VIII-deficient blood were small, with the midpoint of the profiles observed within 1 minute of each other. Thrombin generation during the propagation phase (subsequent to clotting) was greatly impaired in factor VIII deficiency, being depressed to less than 1/29 (<1.9 nmol TAT/L/min) the rate in normal blood (55 nmol TAT/L/min). Replacement with recombinant factor VIII normalized the rate of TAT generation. Thus, coagulation in hemophilia A blood at 25 pmol/L TF is impaired, with significantly slower thrombin generation than normal during the propagation phase; this reduced thrombin appears to affect FPA production and factor V activation more profoundly than platelet activation. At the same level of TF in factor XI-deficient blood (XI:C <2%), only minor differences in clotting or product formation (FPA, osteonectin, and factor Va) were observed. Using reduced levels of initiator (5 pmol/L TF), the reaction was more strongly influenced by factor XI deficiency. Clot formation was delayed from 11.1 to 15.7 minutes, which shortened to 9.7 minutes with factor XI replacement. The maximum thrombin generation rate observed (~37 nmol TAT/L/min) was approximately one third that for normal (110 nmol/L TAT/min) or with factor XI replacement (119 nmol TAT/L/min). FPA release, factor V activation, and release of platelet osteonectin were slower in factor XI-deficient blood than in normal blood. The data demonstrate that factor XI deficiency results in significantly delayed clot formation only at sufficiently low TF concentrations. However, even at these low TF concentrations, significant thrombin is generated in the propagation phase after formation of the initial clot in hemophilia C blood.
- Published
- 1998
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46. The Turnover in Normal Dogs of Prothrombin and Its Fragments; Effect of Induced Intravascular Coagulation
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Owen, Charles A, Mann, Kenneth G, and McDuffie, Frederic C
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- 1979
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47. The Biochemistry of Coagulation
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Mann, Kenneth G.
- Abstract
This article describes the hemostatic process following vascular injury, which involves an integrated response of the blood vessel wall, blood platelets, and plasma b’lood clotting factors, and the way in which these processes relate to one another. A number of diagrams are presented that diagram the component processes.
- Published
- 1984
- Full Text
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48. Immunolocalization of noncollagenous bone matrix proteins in lumbar vertebrae from intact and surgically menopausal cynomolgus monkeys
- Author
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Carlson, Cathy S., Tulli, Hermina M., Jayo, Manuel J., Loeser, Richard F., Tracy, Russell P., Mann, Kenneth G., and Adams, Michael R.
- Abstract
The noncollagenous matrix proteins, composing about 10% of the organic matrix of bone, are considered important for cell matrix organization and regulation of mineralization in bone. In the present study, seven of the major noncollagenous bone matrix proteins were localized immunohistochemically in serial sections of lumbar vertebrae from 24 (12 intact and 12 ovariectomized) adult female cynomolgus monkeys (Macaca fascicularis). Osteocalcin was the only protein restricted to bone cells and mineralized bone matrix. Bone sialoprotein was present in both bone and calcified cartilage, and all the other proteins were distributed in soft tissues as well as bone. Staining for both osteocalcin and bone sialoprotein was present diffusely throughout the bone matrix, but osteonectin, osteopontin, matrix gla protein, decorin, and biglycan staining was concentrated along bone surfaces. Osteoid was negative for osteocalcin and bone sialoprotein, but all other proteins had areas of positive immunostaining within osteoid. All proteins except biglycan exhibited strong immunostaining of a subset of active osteoblasts, suggesting that they may be markers of osteoblast maturity or state of activation. The pattern of immunostaining in intact and surgically menopausal monkeys was similar, except that staining for matrix proteins concentrated along bone surfaces appeared to be more widely distributed in the surgically menopausal monkeys, probably due to the higher rate of bone formation in these animals.
- Published
- 1993
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49. Structure of membrane-bound human factor Va
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Stoylova, Svetla, Mann, Kenneth G., and Brisson, Alain
- Abstract
Coagulation factor Va is an essential cofactor which combines with the serine protease factor Xa on a phospholipid surface to form the prothrombinase complex. In the present study, the structure of factor Va interacting with lipid surfaces containing phosphatidylserine was studied by electron microscopy. Two-dimensional crystals of factor Va were obtained on planar lipid films under quasi-physiological conditions. The two-dimensional projected structure of factor Va was calculated at a resolution of 2 nm, revealing dimers of factor Va arranged on the surface lattice with the symmetry of the plane group p2. Average unit cell dimensions are a= 14.4 nm, b= 8.8 nm, γ = 107°. Each factor Va molecule presents two distinct domains of protein density consisting of one small domain, of 3 nm in diameter, connected to a larger domain of about 6 nm × 4.5 nm. The projected structure of factor Va covers an area equivalent to about fifty phospholipid molecules. In addition, edge-on views of factor Va molecules bound to liposomes reveal a globular structure connected through a thin stem to the liposome surface. A three-dimensional model of membrane-bound factor Va is proposed.
- Published
- 1994
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50. Factor V Quebec Revisited
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Janeway, Claude M., Rivard, Georges E., Tracy, Paula B., and Mann, Kenneth G.
- Abstract
Factor V Quebec has been described as a bleeding disorder that exhibits an autosomal dominant inheritance pattern and presents severe bleeding after trauma. Two members of a fourth-generation (IV. 13 and IV.15) Canadian family have been studied in detail and are the subject of this report. Their clinical presentations and histories have been described previously (Tracy et al: J Clin Invest74:1221, 1984). Persistent abnormalities include mild thrombocytopenia and defective platelet factor V. Plasma factor V is present at near normal concentration and is fully functional. Thus, the bleeding diathesis appears to reflect the absence of platelet factor V activity. The recent report (Hayward et al: Blood84:110a, 1994 [suppl, abstr]) of multimerin deficiency in these individuals led us to reevaluate these patients. Western blot analyses of platelet lysates developed with a variety of monoclonal antibodies show that the α-granule proteins, fibrinogen, von Willebrand factor, factor V and osteonectin are decreased in concentration and significantly degraded in the platelets of these patients. Thrombospondin, while not degraded, is substantially decreased. In contrast, platelet factor 4 and β-thromboglobulin do not appear to be affected. These observations suggest that the α-granules are correctly assembled but the contents are subsequently subjected to proteolytic degradation. The results indicate that factor V Quebec disorder is probably associated with a generalized defect that leads to degradation of most proteins of the α-granules.
- Published
- 1996
- Full Text
- View/download PDF
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