Sample records for antithrombins

  1. Antithrombin deficiency in pregnancy.

    Durai, Shivani; Tan, Lay Kok; Lim, Serene


    We present a case of a 39-year-old, gravida 3 para 2, Chinese female with a history of inherited type 1 Antithrombin deficiency and multiple prior episodes of venous thromboembolism. She presented at 29+4 weeks' gestation with severe pre-eclampsia complicated by haemolysis, elevated liver enzymes and low platelet (HELLP) syndrome. She subsequently underwent an emergency caesarean section for non-reassuring fetal status, which was complicated by postpartum haemorrhage secondary to uterine atony, requiring a B-Lynch suture intraoperatively. PMID:27207982

  2. Antithrombin Dublin (p.Val30Glu)

    Navarro-Fernandez, José; de la Morena-Barrio, María Eugenia; Padilla, José; Miñano, Antonia; Bohdan, Nataliya; Águila, Sonia; Martínez-Martínez, Irene; Sevivas, Teresa S; de Cos, Carmen; Fernández-Mosteirín, Nuria; Llamas, Pilar; Asenjo, Susana; Medina, Pilar; Souto, Juan Carlos; Overvad, Kim; Kristensen, Søren R; Corral, Javier; Vicente, Vicente


    The key haemostatic role of antithrombin and the risk of thrombosis associated with its deficiency support that the low incidence of antithrombin deficiency among patients with thrombosis might be explained by underestimation of this disorder. It was our aim to identify mutations in SERPINC1 caus...

  3. Antithrombin, an Important Inhibitor in Blood Clots.

    Zhu, Ying; Cong, Qing-Wei; Liu, Yue; Wan, Chun-Ling; Yu, Tao; He, Guang; He, Lin; Cai, Lei; Chou, Kuo-Chen


    Blood coagulation is healthy and lifesaving because it can stop bleeding. It can, however, be a troublemaker as well, causing serious medical problems including heart attack and stroke. Body has complex blood coagulation cascade to modulate the blood clots. In the environment of plasma, the blood coagulation cascade is regulated by antithrombin, which is deemed one of the most important serine protease inhibitors. It inhibits thrombin; it can inhibit factors IXa and Xa as well. Interestingly, its inhibitory ability will be significantly increased with the existence of heparin. In this minireview paper, we are to summarize the structural features of antithrombin, as well as its heparin binding modes and anti-coagulation mechanisms, in hopes that the discussion and analysis presented in this paper can stimulate new strategies to find more effective approaches or compounds to modulate the antithrombin. PMID:26411319

  4. Generation of Humanized Mouse Models with Focus on Antithrombin Deficiency

    Jensen, Astrid Bøgh


    Antithrombin is one of the major inhibitors of the coagulation cascade. Antithrombin deficiency in humans significantly increases the risk of thrombosis. Mice homozygous for an antithrombin knock out die at day 16.5 of gestation, due to disseminated intravascular coagulation and organ failure. I ...

  5. Neuraxial anesthesia for labor and cesarean delivery in a parturient with hereditary antithrombin deficiency on recombinant human antithrombin infusion therapy.

    Pamnani, Anup; Rosenstein, Megan; Darwich, Alaeldin; Wolfson, Alexander


    A recombinant human antithrombin (rhAT; generic name: antithrombin Alfa) has recently been developed. A 37 year-old parturient with hereditary antithrombin deficiency, receiving rhAT infusion therapy, who successfully received an epidural catheter for analgesia and anesthesia during labor and cesarean delivery, is presented. PMID:20868967

  6. Antithrombin III for critically ill patients

    Afshari, Arash; Wetterslev, Jørn; Brok, Jesper Sune;


    Critical illness is associated with uncontrolled inflammation and vascular damage which can result in multiple organ failure and death. Antithrombin III (AT III) is an anticoagulant with anti-inflammatory properties but the efficacy and any harmful effects of AT III supplementation in critically ...

  7. Antithrombin III Basel. Identification of a Pro-Leu substitution in a hereditary abnormal antithrombin with impaired heparin cofactor activity.

    Chang, J Y; Tran, T H


    Antithrombin III Basel is a hereditary abnormal antithrombin with normal progressive inhibition activity (normal reactive site) and reduced heparin cofactor activity (impaired heparin binding site). Structures of antithrombin III Basel and normal antithrombin III isolated from the same patient were compared by peptide mapping using the dimethylaminoazobenzene isothiocyanate precolumn derivatization technique. Of the approximately 50 tryptic peptides of normal and abnormal antithrombin III, one peptide comprising residues 40-46 had a different retention time in reversed-phase high performance liquid chromatography. The amino acid sequence of the peptide from antithrombin III Basel had a single substitution of Pro (normal) by Leu (abnormal) at position 41. This substitution is close to an Arg (residue 47) and a Trp (residue 49) which have previously been shown to be critical for heparin binding by antithrombin III. Although additional amino acid substitutions in antithrombin III Basel cannot be ruled out, this Pro-Leu replacement could cause a conformational change by increasing both the helical structure and the hydrophobicity around residue 41. These data suggest that: (i) the heparin binding site of antithrombin III encompasses the region containing residues 41, 47, and 49; and (ii) the impaired heparin cofactor activity of antithrombin III Basel is likely due to a conformational change of the heparin binding site induced by the Pro-Leu substitution at position 41. PMID:3080419

  8. 21 CFR 864.7060 - Antithrombin III assay.


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antithrombin III assay. 864.7060 Section 864.7060 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... assay. (a) Identification. An antithrombin III assay is a device that is used to determine the...

  9. Sulfated cellulose thin films with antithrombin affinity


    Full Text Available Cellulose thin films were chemically modified by in situ sulfation to produce surfaces with anticoagulant characteristics. Two celluloses differing in their degree of polymerization (DP: CEL I (DP 215–240 and CEL II (DP 1300–1400 were tethered to maleic anhydride copolymer (MA layers and subsequently exposed to SO3•NMe3 solutions at elevated temperature. The impact of the resulting sulfation on the physicochemical properties of the cellulose films was investigated with respect to film thickness, atomic composition, wettability and roughness. The sulfation was optimized to gain a maximal surface concentration of sulfate groups. The scavenging of antithrombin (AT by the surfaces was determined to conclude on their potential anticoagulant properties.

  10. Antithrombin in the treatment of burn trauma.

    Kowal-Vern, Areta; Orkin, Bruce A


    Antithrombin (AT) is a natural anticoagulant with anti-inflammatory properties that has demonstrated value in sepsis, disseminated intravascular coagulation and in burn and inhalation injury. With high doses, AT may decrease blood loss during eschar excision, reducing blood transfusion requirements. There are no human randomized, placebo-controlled studies, which have tested the true benefit of this agent in these conditions. Two main forms of AT are either plasma-derived AT (phAT) and recombinant AT (rhAT). Major ovine studies in burn and smoke inhalation injury have utilized rhAT. There have been no studies which have either translated the basic rhAT research in burn trauma, or determined the tolerance and pharmacokinetics of rhAT concentrate infusions in burn patients. Advantages of rhAT infusions are no risk of blood borne diseases and lower cost. However, the majority of human burn patient studies have been conducted utilizing phAT. Recent Japanese clinical trials have started using phAT in abdominal sepsis successfully. This review examines the properties of both phAT and rhAT, and analyzes studies in which they have been utilized. We believe that it is time to embark on a randomized placebo-controlled multi-center trial to establish the role of AT in both civilian and military patients with burn trauma. PMID:26855890

  11. Antithrombin III for critically ill patients

    Allingstrup, Mikkel; Wetterslev, Jørn; Ravn, Frederikke B;


    BACKGROUND: Critical illness is associated with uncontrolled inflammation and vascular damage which can result in multiple organ failure and death. Antithrombin III (AT III) is an anticoagulant with anti-inflammatory properties but the efficacy and any harmful effects of AT III supplementation in......, moderate quality of evidence) in the AT III group. The amount of red blood cells administered had a mean difference (MD) of 138.49 (95% Cl -391.35 to 668.34, I² statistic = 84%, random-effect model, 4 trials, 137 participants, very low quality of evidence). The effect of AT III in patients with multiple...... organ failure (MOF) was a MD of -1.24 (95% Cl -2.18 to -0.29, I² statistic = 48%, random-effects model, 3 trials, 156 participants, very low quality of evidence) and for patients with an Acute Physiology and Chronic Health Evaluation score (APACHE) at II and III the MD was -2.18 (95% Cl -4.36 to -0...

  12. Magnetic particles as affinity matrix for purification of antithrombin

    Mercês, A. A. D.; Maciel, J. C.; Carvalho Júnior, L. B.


    Immobilization of biomolecules onto insoluble supports is an important tool for the fabrication of a diverse range of functional materials. It provides advantages: enhanced stability and easy separation. In this work two different magnetic composites were synthesized (MAG-PANI-HS and mDAC-HS) to human antithrombin purification. The magnetic particles (MAG) were obtained by co-precipitation method of iron salts II and III and subsequently coated with polyaniline (MAG-PANI particles). Dacron (polyethylene terephthalate) suffered a hydrazinolysis reaction to obtain a powder (Dacron hydrazide) which was subsequently magnetized (mDAC particles) also by co-precipitation method. Heparan sulfate (HS) was immobilized to MAG-PANI and mDAC retained respectively 35μg and 38.6μg per of support. The magnetic composite containing HS immobilized (MAG-PANI-HS and mDAC-HS) was incubated with human blood plasma (1mL) and then washed with NaCl gradients. Electrophoresis of proteins present in eluates showed bands of antithrombin (58kDa). A reduction in the antithrombin activity was detected in plasma that were incubated in the composites magnetic with HS immobilized, suggesting that the antithrombin was removed of the human blood plasma and then purified. Therefore, the above results suggest that both preparations: MAG-PANI-HS and mDAC-HS are able to affinity purify antithrombin, an important component of blood coagulation.

  13. Antithrombin III for critically ill patients

    Allingstrup, Mikkel; Wetterslev, Jørn; Ravn, Frederikke B.;


    PURPOSE: Antithrombin III (AT III) is an anticoagulant with anti-inflammatory properties. We assessed the benefits and harms of AT III in critically ill patients. METHODS: We searched from inception to 27 August 2015 in CENTRAL, MEDLINE, EMBASE, CAB, BIOSIS and CINAHL. We included randomized cont...

  14. Use of recombinant human antithrombin concentrate in pregnancy

    Baumann Kreuziger LM


    Full Text Available Lisa M Baumann Kreuziger,1 Tracy L Prosen,2 Mark T Reding1 1Division of Hematology, Oncology, and Transplantation, University of Minnesota, MN, USA; 2Obstetrics and Gynecology, Maternal Fetal Medicine Center, University of Minnesota, MN, USAWe read with great interest James et al’s article "Prevention and treatment of venous thromboembolism in pregnancy in patients with hereditary antithrombin deficiency." The authors reported a case series of six women with antithrombin (AT deficiency treated with plasma derived antithrombin concentrate (pdAT; Thrombate III®, Grifols Therapeutics, Clayton, NC, USA.1 In contrast to these cases, we have managed three AT-deficient women over the past two years with low-molecular-weight-heparin during pregnancy and recombinant human antithrombin concentrate (rhAT; ATryn®, GTC Biotherapeutics, Framingham, MA, USA at delivery. In the Phase III trial leading to approval of rhAT, pregnant women required frequent dose modifications, and an alternate dosing regimen is currently recommended by the manufacturer.2,3 The detailed information provided in these cases may assist future providers in managing pregnant patients with AT deficiency.View original paper by James and colleagues.

  15. Inherited antithrombin deficiency and anabolic steroids: a risky combination.

    Choe, Hannah; Elfil, Mohamed; DeSancho, Maria T


    A 20-year-old male with asymptomatic inherited type 1 antithrombin deficiency and a family history of thrombosis started injecting himself with testosterone 250 mg intramuscularly twice weekly for 5 weeks. He presented to the hospital with progressive dyspnea on exertion, chest pain and hemoptysis. Workup revealed bilateral submassive pulmonary embolism and proximal right lower extremity deep vein thrombosis. He was treated with intravenous (IV) unfractionated heparin and underwent catheter-directed thrombolysis with alteplase to the main pulmonary arteries. Postprocedure, he remained on IV alteplase infusion for 24 h and unfractionated heparin in the intensive care unit. Concomitantly he received plasma-derived antithrombin concentrate. He was transitioned to subcutaneous enoxaparin twice daily and discharged from the hospital on oral rivaroxaban 15 mg twice a day. This case highlights the heightened thrombogenic effect of anabolic steroids in the setting of underlying thrombophilia especially in younger subjects. PMID:26588446

  16. Interaction of antithrombin III with preadsorbed albumin-heparin conjugates

    Hennink, W.E.; Ebert, C.D.; Kim, S. W.; Breemhaar, W.; Bantjes, A.; Feijen, J.


    The adsorption of antithrombin III (AT III) onto polystyrene surfaces preadsorbed with albumin or albuminheparin conjugates was studied using a two step enzyme immuno assay. When AT III-buffer solutions were used, the highest adsorption values were measured on high affinity albumin-heparin conjugate pretreated surfaces. Less AT III adsorption was found on nonfractionated albumin-heparin conjugate preadsorbed surfaces. AT III adsorption could also be detected on low affinity conjugate and albu...

  17. Antithrombin controls tumor migration, invasion and angiogenesis by inhibition of enteropeptidase

    Luengo-Gil, Ginés; Calvo, María Inmaculada; Martín-Villar, Ester; Águila, Sonia; Bohdan, Nataliya; Antón, Ana I.; Espín, Salvador; Ayala de la Peña, Francisco; Vicente, Vicente; Corral, Javier; Quintanilla, Miguel; Martínez-Martínez, Irene


    Antithrombin is a key inhibitor of the coagulation cascade, but it may also function as an anti-inflammatory, anti-angiogenic, anti-viral and anti-apoptotic protein. Here, we report a novel function of antithrombin as a modulator of tumor cell migration and invasion. Antithrombin inhibited enteropeptidase on the membrane surface of HT-29, A549 and U-87 MG cells. The inhibitory process required the activation of antithrombin by heparin, and the reactive center loop and the heparin binding domain were essential. Surprisingly, antithrombin non-covalently inhibited enteropeptidase, revealing a novel mechanism of inhibition for this serpin. Moreover, as a consequence of this inhibition, antithrombin was cleaved, resulting in a molecule with anti-angiogenic properties that reduced vessel-like formation of endothelial cells. The addition of antithrombin and heparin to U-87 MG and A549 cells reduced motility in wound healing assays, inhibited the invasion in transwell assays and the degradation of a gelatin matrix mediated by invadopodia. These processes were controlled by enteropeptidase, as demonstrated by RNA interference experiments. Carcinoma cell xenografts in nude mice showed in vivo co-localization of enteropeptidase and antithrombin. Finally, treatment with heparin reduced experimental metastasis induced by HT29 cells in vivo. In conclusion, the inhibition of enteropeptidase by antithrombin may have a double anti-tumor effect through inhibiting a protease involved in metastasis and generating an anti-angiogenic molecule. PMID:27270881

  18. Discordant diagnoses obtained by different approaches in antithrombin mutation analysis

    Feddersen, Søren; Nybo, Mads

    OBJECTIVES: In hereditary antithrombin (AT) deficiency it is important to determine the underlying mutation since the future risk of thromboembolism varies considerably between mutations. DNA investigations are in general thought of as flawless and irrevocable, but the diagnostic approach can be...... critical. We therefore investigated mutation results in the AT gene, SERPINC1, with two different approaches. DESIGN AND METHODS: Sixteen patients referred to the Centre for Thrombosis and Haemostasis, Odense University Hospital, with biochemical indications of AT deficiency, but with a negative denaturing...... high-performance liquid chromatography (DHPLC) mutation screening (routine approach until recently) were included. As an alternative mutation analysis, direct sequencing of all exons and exon-intron boundaries without pre-selection by DHPLC was performed. RESULTS: Out of sixteen patients with a...

  19. Antithrombin gene Arg197Stop mutation-associated venous sinus thrombosis in a Chinese family

    Ang Li; Dexin Wang; Qiming Xue; Baoen Wang; Tianhui Liu; Zhandong Liu; Jimei Li; Chunling Zhang; Jun Chen; Jinmei Sun; YanfeiHan; Lili Wang


    This study sought to elucidate the genetic correlation of cerebral venous sinus thrombosis caused by a hereditary antithrombin deficiency in a Chinese family, at the genetic and protein levels. A nonsense mutation from C to T on locus 6431 in exon 3B of the antithrombin gene was observed,leading to an arginine (CGA) to stop codon (TGA) change in the protein. This is the first report of this mutation in China. Ineffective heparin therapy in the propositus patient is associated with a lack of heparin binding sites after antithrombin gene mutation. Characteristic low intracranial pressure in the acute phase might be specific to this patient with cerebral venous sinus thrombosis.

  20. Antithrombin can modulate coagulation, cytokine production, and expression of adhesion molecules in abdominal aortic aneurysm repair surgery.

    Nishiyama, Tomoki


    We investigated the effects of antithrombin on coagulation, fibrinolysis, and production of cytokines and adhesion molecules in abdominal aortic aneurysm repair surgery. Sixteen patients for Y-shaped graft replacement of abdominal aortic aneurysm were divided into an antithrombin group and a control group. In the antithrombin group, 3000 U antithrombin was infused over 30 min before heparin administration and 24 h later. White blood cell counts, platelet counts, prothrombin time ratio, and serum concentrations of antithrombin, polymorphonuclear leukocyte elastase, interleukin (IL)-1beta, IL-6, IL-8, tumor necrosis factor-alpha, and adhesion molecules, and variables of coagulation and fibrinolysis were measured before surgery, at the end of surgery, and 1 and 2 days after surgery. The antithrombin concentration decreased in the control group, whereas it increased in the antithrombin group with significant differences between the groups. Prothrombin time ratio, concentrations of d-dimer, thrombin-antithrombin complex, and intercellular adhesion molecule-1 increased only in the control group and polymorphonuclear leukocyte elastase, IL-6, tumor necrosis factor-alpha, and vascular cell adhesion molecule-1 increased in both groups. They were significantly less in the antithrombin group except for intercellular adhesion molecule-1. In conclusion, antithrombin could decrease hypercoagulation and inflammatory activation during abdominal aortic aneurysm surgery, which may decrease adverse events. PMID:16551889

  1. Activation of antithrombin III isoforms by heparan sulphate glycosaminoglycans and other sulphated polysaccharides.

    Carlson, T H; Kolman, M R; Piepkorn, M


    Antithrombin III occurs naturally as two functionally distinct molecular species that differ in glycosylation at Asn135. Whereas the predominant, glycosylated isoform has high affinity for heparin, a quantitatively minor isoform lacking glycosylation at that site displays relatively higher affinity for both heparins and heparinoids. We characterized the ability of various sulphated polysaccharides to potentiate the rates of thrombin inhibition by the isoforms. High-molecular-weight dextran sulphate was the most effective of those studied, increasing thrombin inhibition by the higher-affinity antithrombin III isoform up to five-fold more efficiently than did heparin fractions with low-affinity for antithrombin III. In addition, dextran sulphate activated the higher-affinity isoform as much as twelve times more effectively than it did the lower-affinity isoform. Pentosan polysulphate was up to three-fold, and some heparan sulphate fractions up to two-fold, more effective with the higher, compared with the lower affinity, isoform. Heparan sulphate preparations less effectively increased the rate of thrombin inhibition than did the other low-affinity polysaccharides. Structure-function studies indicated positive correlations between activity and both polymer length and anionic group density of low-affinity sulphated polysaccharides. The observed effects of the heparan sulphates on this anticoagulant pathway, although of low potency, are consistent with the hypotheses that these substances naturally regulate blood homeostasis in vascular tissues and that much of this function may be mediated by the higher-affinity antithrombin III isoform. PMID:8589216

  2. Anticoagulant and anti-inflammatory effects after peritoneal lavage with antithrombin in experimental polymicrobial peritonitis

    S.Q. van Veen; C.W. Cheung; J.C.M. Meijers; T.M. van Gulik; M.A. Boermeester


    Background: In sepsis, coagulation inhibition using high-dose systemic antithrombin (AT) tends to improve survival. However, systemic AT use is complicated by increased risk of bleeding (odds ratio 1,7) and clinically important survival increase is seen only in the non-heparinized subgroup. Local (i

  3. Antithrombin inhibits bronchoalveolar activation of coagulation and limits lung injury during Streptococcus pneumoniae pneumonia in rats

    Choi, Goda; Hofstra, Jorrit-Jan H; Roelofs, Joris J T H; Rijneveld, Anita W; Bresser, Paul; van der Zee, Jaring S; Florquin, Sandrine; van der Poll, Tom; Levi, Marcel; Schultz, Marcus J


    OBJECTIVE: Alveolar fibrin deposition is a hallmark of pneumonia. It has been proposed that natural inhibitors of coagulation, including activated protein C, antithrombin, and tissue factor pathway inhibitor, exert lung-protective effects via anticoagulant and possibly anti-inflammatory pathways. We

  4. Regions flanking exon 1 regulate constitutive expression of the human antithrombin gene.

    Fernandez-Rachubinski, F A; Weiner, J H; Blajchman, M A


    We have identified cis-acting elements and trans-acting factors that regulate constitutive expression of the human antithrombin gene. The activity of the sequences flanking the first exon of the gene was investigated using a luciferase-based reporter assay in transiently transfected HepG2, COS1, BSC40, and HeLa cells. Deletion analysis allowed the mapping of two elements able to promote antithrombin gene transcription in HepG2 and COS1 cells. The first element is located upstream of the first exon (-150/+68 nucleotides). The second element is in the first intervening sequence (+300/+700 nucleotides) and functions in an orientation opposite to that of the first. Footprint analysis showed three protected areas in the 5' upstream element at -92/-68 (element A), -14/+37 (element B), and -126/-100 nucleotides (element C). These elements acted as enhancers in luciferase reporter assays. Gel retardation analysis demonstrated that two liver-enriched transcription factors, hepatocyte nuclear factor 4 (HNF4) and CCAAT enhancer-binding protein (C/EBPa), bound to the 5' upstream element. HNF4 bound to elements A and C, whereas C/EBPa bound to element B. Element A also interacted with the ubiquitous nuclear hormone receptors chicken ovalbumin upstream promoter transcription factor 1 (COUP-TF1), thyroid hormone receptor alpha (TRalpha), peroxisome proliferator-activated receptor alpha(PPARalpha), and retinoid X receptor alpha (RXRalpha). In HepG2 and BSC40 cells, HNF4, C/EBPalpha, and RXRalpha activated luciferase expression from a reporter construct containing the 5'-upstream minimal antithrombin gene promoter, while COUP-TF1, TRalpha, and HNF3 (alpha or beta) repressed such expression. Our results show that constitutive expression of the human antithrombin gene depends in part upon the interplay of these transcription factors and suggest that signaling pathways regulated by these factors can modulate antithrombin gene transcription. PMID:8910619

  5. Antithrombin significantly influences platelet adhesion onto immobilized fibrinogen in an in-vitro system simulating low flow

    Scharf Rüdiger E


    Full Text Available Abstract Background Adhesion of platelets onto immobilized fibrinogen is of importance in initiation and development of thrombosis. According to a recent increase in evidence of a multiple biological property of antithrombin, we evaluated the influence of antithrombin on platelet adhesion onto immobilized fibrinogen using an in-vitro flow system. Methods Platelets in anticoagulated whole blood (29 healthy blood donors were labelled with fluorescence dye and perfused through a rectangular flow chamber (shear rates of 13 s-1 to 1500 s-1. Platelet adhesion onto fibrinogen-coated slips was assessed using a fluorescence laser-scan microscope and compared to the plasma antithrombin activity. Additionally the effect of supraphysiological AT supplementation on platelets adhesion rate was evaluated. Results Within a first minute of perfusion, an inverse correlation between platelet adhesion and plasma antithrombin were observed at 13 s-1 and 50 s-1 (r = -0.48 and r = -0.7, p -1, within first minute have been found. An in-vitro supplementation of whole blood with antithrombin increased the antithrombin activity up to 280% and platelet adhesion rate reached about 65% related to the adhesion rate in a non-supplemented blood (1.25 ± 0.17 vs. 1.95 ± 0.4 p = 0.008, respectively. Conclusion It appears that antithrombin in a low flow system suppresses platelet adhesion onto immobilized fibrinogen independently from its antithrombin activity. A supraphysiological substitution of blood with antithrombin significantly reduces platelet adhesion rate. This inhibitory effect might be of clinical relevance.

  6. Identification of antithrombin-modulating genes. Role of LARGE, a gene encoding a bifunctional glycosyltransferase, in the secretion of proteins?

    María Eugenia de la Morena-Barrio

    Full Text Available The haemostatic relevance of antithrombin together with the low genetic variability of SERPINC1, and the high heritability of plasma levels encourage the search for modulating genes. We used a hypothesis-free approach to identify these genes, evaluating associations between plasma antithrombin and 307,984 polymorphisms in the GAIT study (352 individuals from 21 Spanish families. Despite no SNP reaching the genome wide significance threshold, we verified milder positive associations in 307 blood donors from a different cohort. This validation study suggested LARGE, a gene encoding a protein with xylosyltransferase and glucuronyltransferase activities that forms heparin-like linear polysaccharides, as a potential modulator of antithrombin based on the significant association of one SNPs, rs762057, with anti-FXa activity, particularly after adjustment for age, sex and SERPINC1 rs2227589 genotype, all factors influencing antithrombin levels (p = 0.02. Additional results sustained this association. LARGE silencing inHepG2 and HEK-EBNA cells did not affect SERPINC1 mRNA levels but significantly reduced the secretion of antithrombin with moderate intracellular retention. Milder effects were observed on α1-antitrypsin, prothrombin and transferrin. Our study suggests LARGE as the first known modifier of plasma antithrombin, and proposes a new role for LARGE in modulating extracellular secretion of certain glycoproteins.

  7. Mechanism of poly(acrylic acid) acceleration of antithrombin inhibition of thrombin: implications for the design of novel heparin mimics.

    Monien, Bernhard H; Cheang, Kai I; Desai, Umesh R


    The bridging mechanism of antithrombin inhibition of thrombin is a dominant mechanism contributing a massive approximately 2500-fold acceleration in the reaction rate and is also a key reason for the clinical usage of heparin. Our recent study of the antithrombin-activating properties of a carboxylic acid-based polymer, poly(acrylic acid) (PAA), demonstrated a surprisingly high acceleration in thrombin inhibition (Monien, B. H.; Desai, U. R. J. Med. Chem. 2005, 48, 1269). To better understand this interesting phenomenon, we have studied the mechanism of PAA-dependent acceleration in antithrombin inhibition of thrombin. Competitive binding studies with low-affinity heparin and a heparin tetrasaccharide suggest that PAA binds antithrombin in both the pentasaccharide- and the extended heparin-binding sites, and these results are corroborated by molecular modeling. The salt-dependence of the K(D) of the PAA-antithrombin interaction shows the formation of five ionic interactions. In contrast, the contribution of nonionic forces is miniscule, resulting in an interaction that is significantly weaker than that observed for heparins. A bell-shaped profile of the observed rate constant for antithrombin inhibition of thrombin as a function of PAA concentration was observed, suggesting that inhibition proceeds through the "bridging" mechanism. The knowledge gained in this mechanistic study highlights important rules for the rational design of orally available heparin mimics. PMID:16078853

  8. Domain structure of antithrombin III. Tentative localization of the heparin binding region using 1H NMR spectroscopy

    The denaturation of human and bovine antithrombin III by guanidine hydrochloride has been followed by 1H NMR spectroscopy. The same unfolding transition seen previously from circular dichroism studies at low denaturant concentration was detected here by discontinuous changes in the chemical shifts of the C(2) protons of two of the five histidines in human antithrombin III and of three of the six histidines in bovine antithrombin III. These two histidines in human antithrombin III are assigned to residue 1 and, more tentatively, to residue 65. Two of the three histidines similarly affected in the bovine protein appear to be homologous to residues in the human protein. This supports the proposal of similar structures for the two proteins. In the presence of heparin, the discontinuous titration behavior of these histidine resonances is shifted to higher denaturant concentration, reflecting the stabilization of the easily unfolded first domain of the protein by bound heparin. From the tentative assignment of one of these resonances to histidine-1, it is proposed that the heparin binding site of antithrombin III is located in the N-terminal region and that this region forms a separate domain from the rest of the protein. The pattern of disulfide linkages is such that this domain may well extend from residue 1 to at least residue 128. Thermal denaturation also leads to major perturbation of these two histidine resonances in human antithrombin III, though stable intermediates in the unfolding were not detected

  9. Increased N-glycosylation efficiency by generation of an aromatic sequon on N135 of antithrombin.

    Sonia Aguila

    Full Text Available The inefficient glycosylation of consensus sequence on N135 in antithrombin explains the two glycoforms of this key anticoagulant serpin found in plasma: α and β, with four and three N-glycans, respectively. The lack of this N-glycan increases the heparin affinity of the β-glycoform. Recent studies have demonstrated that an aromatic sequon (Phe-Y-Asn-X-Thr in reverse β-turns enhances N-glycosylation efficiency and stability of different proteins. We evaluated the effect of the aromatic sequon in this defective glycosylation site of antithrombin, despite of being located in a loop between the helix D and the strand 2A. We analyzed the biochemical and functional features of variants generated in a recombinant cell system (HEK-EBNA. Cells transfected with wild-type plasmid (K133-Y-N135-X-S137 generated 50% of α and β-antithrombin. The S137T, as previously reported, K133F, and the double mutant (K133F/S137T had improved glycosylation efficiency, leading to the secretion of α-antithrombin, as shown by electrophoretic and mass analysis. The presence of the aromatic sequon did not significantly affect the stability of this conformationally sensitive serpin, as revealed by thermal denaturation assay. Moreover, the aromatic sequon hindered the activation induced by heparin, in which is involved the helix D. Accordingly, K133F and particularly K133F/S137T mutants had a reduced anticoagulant activity. Our data support that aromatic sequons in a different structural context from reverse turns might also improve the efficiency of N-glycosylation.

  10. Rapid high-performance liquid chromatographic quantification of recombinant human antithrombin III during production and purification

    Büntemeyer, Heino; Tebbe, H.; Lütkemeyer, Dirk; Lehmann, Jürgen


    For monitoring of recombinant human antithrombin III during cell culture processes and subsequent purification steps a rapid method for quantitative determination was developed. The need for the introduction of this rapid method came from the limited availability of a quantitative enzyme-linked immunosorbent assay (ELISA) and the very time-consuming ELISA procedure. The developed method is based on reversed-phase high-performance liquid chromatography using a C 4 column. The separation by gra...

  11. An Antithrombin-Heparin Complex Increases the Anticoagulant Activity of Fibrin Clots

    Lesley J. Smith


    Full Text Available Clotting blood contains fibrin-bound thrombin, which is a major source of procoagulant activity leading to clot extension and further activation of coagulation. When bound to fibrin, thrombin is protected from inhibition by antithrombin (AT + heparin but is neutralized when AT and heparin are covalently linked (ATH. Here, we report the surprising observation that, rather than yielding an inert complex, thrombin-ATH formation converts clots into anticoagulant surfaces that effectively catalyze inhibition of thrombin in the surrounding environment.

  12. Anti-thrombin III, Protein C, and Protein S deficiency in acute coronary syndrome

    Dasnan Ismail


    Full Text Available The final most common pathway for the majority of coronary artery disease is occlusion of a coronary vessel. Under normal conditions, antithrombin III (AT III, protein C, and protein S as an active protein C cofactor, are natural anticoagulants (hemostatic control that balances procoagulant activity (thrombin antithrombin complex balance to prevent thrombosis. If the condition becomes unbalanced, natural anticoagulants and the procoagulants can lead to thrombosis. Thirty subjects with acute coronary syndrome (ACS were studied for the incidence of antithrombin III (AT III, protein C, and protein S deficiencies, and the result were compare to the control group. Among patients with ACS, the frequency of distribution of AT-III with activity < 75% were 23,3% (7 of 30, and only 6,7% ( 2 of 30 in control subject. No one of the 30 control subject have protein C activity deficient, in ACS with activity < 70% were 13,3% (4 of 30. Fifteen out of the 30 (50% control subjects had protein S activity deficiency, while protein S deficiency activity < 70% was found 73.3.% (22 out of 30. On linear regression, the deterministic coefficient of AT-III activity deficiency to the development ACS was 13,25 %, and the deterministic coefficient of protein C activity deficient to the development of ACS was 9,06 %. The cut-off point for AT-III without protein S deficiency expected to contribute to the development of vessel disease was 45%. On discriminant analysis, protein C activity deficiency posed a risk for ACS of 4,5 greater than non deficient subjects, and AT-III activity deficiency posed a risk for ACS of 3,5 times greater than non deficient subjects. On binary logistic regression, protein S activity acted only as a reinforcing factor of AT-III activity deficiency in the development of ACS. Protein C and AT III deficiency can trigger ACS, with determinant coefficients of 9,06% and 13,25% respectively. Low levels of protein C posed a greater risk of

  13. Comparison of antithrombin activity of the polysulphate chitosan derivatives in in vivo and in vitro system.

    Drozd, N N; Sher, A I; Makarov, V A; Galbraikh, L S; Vikhoreva, G A; Gorbachiova, I N


    In order to choose the proper method for evaluating the antithrombin activity in samples of chitosan polysulphate (CP) with different polymerization degrees and sulphation degrees, we estimated the ability of direct anticoagulants to depress the coagulability of recalcified sheep blood using the third international heparin standard (A1 - in vitro system) and determined such activity on pharmacodynamic curve (A2 - in vivo system). The curve admits the kinetics of CP elimination to be nonlinear in case of intravenous injection to rabbits, as it is observed in heparin: Ct = C(o)exp(-K(e)lt), where Ct is the CP concentration at the time moment t; C(o) is the CP concentration at the injection moment; Kel is the elimination constant. Besides, it is assumed that there is a linear approximation of the anticoagulant effect on the dose, which finally makes it possible to calculate the specific activity A2: T = KTCt+T(in), where T is the time of clot formation at different time intervals after CP injection; T(in) is the time of clot formation prior to CP injection. T value was assessed in two tests: blood coagulation time (BCT) and activated partial thromboplastin time (APTT). No correlation was observed between A1 and A2. At the same time, the values of Kel and the period of semi-elimination, with the use of the biospecific cetylpyridinium chloride electrophores for the quantitative determination of CP in rabbit's blood taken at different time intervals after injection, showed a close correlation (r = .94, P < .05) between the same parameters, obtained with the help of the rectilinear pharmacodynamic plot in BCT test. Thus, experimentally, it was proven that the assumption of the CP nonlinear elimination and the CP effect-dose dependence was true, which is necessary for A2 calculation. Relatively low molecular weights (MW 61-82 kDa, polymerization degree 188-252 ) and high sulphation patterns (sulphur amounts 15.6-16.9%, sulphation degree 1.58-1.86) were slowly cleared and

  14. Histidine-rich glycoprotein does not interfere with interactions between antithrombin III and heparin-like compounds on vascular endothelial cells

    The role of histidine-rich glycoprotein in controlling heparin-like compounds on the endothelial cell surface is still unclear. The effects of this heparin-neutralizing protein on the interaction between antithrombin III and cultured porcine aortic endothelial cells were examined. Displacement of 125I-labeled antithrombin III specifically bound to endothelial cells by unlabeled histidine-rich glycoprotein was much less potent than that by unlabeled antithrombin III. One hundred-fold molar excess of histidine-rich glycoprotein displaced specific 125I-antithrombin III binding only by 20%. Furthermore, the endothelial cell-mediated acceleration of thrombin inactivation by antithrombin III was diminished by protamine sulfate, but was not affected by histidine-rich glycoprotein even at a histidine-rich glycoprotein/antithrombin III molar ratio of approximately 7:1. These data indicate that histidine-rich glycoprotein does not interfere with the interaction of endothelial cell heparin-like compounds with antithrombin III. Thus, it may not play an important role in the modulation of anticoagulant activity of endothelial cells in vivo, suggesting that the commonly accepted view of the probable function of this protein is erroneous

  15. Dynamic properties of the native free antithrombin from molecular dynamics simulations: computational evidence for solvent- exposed Arg393 side chain.

    Tóth, László; Fekete, Attila; Balogh, Gábor; Bereczky, Zsuzsanna; Komáromi, István


    While antithrombin (AT) has small basal inhibitory activity, it reaches its full inhibitory potential against activated blood coagulation factors, FXa, FIXa, and FIIa (thrombin), via an allosteric and/or template (bridging) mechanism by the action of heparin, heparan sulfate, or heparin-mimetic pentasaccharides (PS). From the numerous X-ray structures available for different conformational states of AT, only indirect and incomplete conclusions can be drawn on the inherently dynamic properties of AT. As a typical example, the basal inhibitory activity of AT cannot be interpreted on the basis of "non-activated" free antithrombin X-ray structures since the Arg393 side chain, playing crucial role in antithrombin-proteinase interaction, is not exposed. In order to reveal the intrinsic dynamic properties and the reason of basal inhibitory activity of antithrombin, 2 μs molecular dynamics simulations were carried out on its native free-forms. It was shown from the simulation trajectories that the reactive center loop which is functioning as "bait" for proteases, even without any biasing potential can populate conformational state in which the Arg393 side chain is solvent exposed. It is revealed from the trajectory analysis that the peptide sequences correspond to the helix D extension, and new helix P formation can be featured with especially large root-mean-square fluctuations. Mutual information analyses of the trajectory showed remarkable (generalized) correlation between those regions of antithrombin which changed their conformations as the consequence of AT-PS complex formation. This suggests that allosteric information propagation pathways are present even in the non-activated native form of AT. PMID:25483839

  16. Polyurethane films modified by antithrombin-heparin complex to enhance endothelialization: An original impedimetric analysis

    In this paper, polyurethane (PU) was deposited as a thin layer onto the surface of ITO (indium tin oxide) and was then modified with an antithrombin-heparin complex (ATH). The resulting films were characterized by ATR spectroscopy, contact angle measurements and electrochemical impedance spectroscopy (EIS). Physicochemical characterization confirmed the surface modifications. The obtained films were used as substrates for endothelial cell attachment and growth. These processes were characterized using electrochemical impedance spectroscopy (EIS). We observed that the addition of a small amount of heparin and AT additives onto the polymer surface resulted in a considerable change in the surface characteristics, and we found that PU films that were modified by the ATH complex were able to greatly enhance adhesion and proliferation of endothelial cells (ECs).

  17. Antithrombin attenuates myocardial dysfunction and reverses systemic fluid accumulation following burn and smoke inhalation injury: a randomized, controlled, experimental study

    Rehberg, S; Yamamoto, Y; Bartha, E; Sousse, L.E. (Linda); Jonkam, C. (Collette); Zhu, Y; Traber, L.D. (Lillian); Cox, R. A.; Traber, D.L. (Daniel); Enkhbaatar, P


    Introduction: We hypothesized that maintaining physiological plasma levels of antithrombin attenuates myocardial dysfunction and inflammation as well as vascular leakage associated with burn and smoke inhalation injury. Therefore, the present prospective, randomized experiment was conducted using an established ovine model. Methods: Following 40% of total body surface area, third degree flame burn and 4 × 12 breaths of cold cotton smoke, chronically instrumented sheep were randomly assigned t...

  18. A recurrent deletion in the antithrombin gene, AT106-108(-6 bp), identified by DNA heteroduplex detection

    Olds, R.J.; Thein, S.L. (John Radcliffe Hospital, Oxford (United Kingdom)); Lane, D.A. (Charing Cross and Westminster Medical School, London (United Kingdom)); Beresford, C.H.; Hughes, P.M. (Univ. of Otago Medical School, Dunedin (New Zealand)); Abildgaard, U. (Aker Hospital, Oslo (Norway))


    Antithrombin is the major physiological inhibitor of the activated serine proteinases of the coagulation system. Hereditary deficiency of the inhibitor is transmitted in an autosomal dominant pattern and is associated with a risk of venous thromboembotic disease in affected individuals. In the classical form of deficiency, type Ia, plasma antithrombin is reduced to approximately half normal in both functional and immunological assays. The authors report here the identification of a recurrent mutation as the basis for type Ia deficiency in two independent kindreds, one from New Zealand and the other from Norway, and demonstrate the utility of DNA heteroduplex detection as a method for screening for the presence of mutations. Standard functional and immunological assays for plasma antithrombin showed levels of approximately half normal in several members of both kindreds, consistent with the classification as type Ia deficiency. The plasma of the proband from the Norwegian kindred was examined by crossed immunoelectrophoresis, in the presence or absence of heparin in the first dimension, and an abnormal component that may have represented a variant form of the inhibitor was not identified. In both families affected members have had episodes of venous thrombosis, although some carriers of the abnormal allele, as confirmed in the current study, so far have not had clinical thrombotic disease. 7 refs., 2 figs.

  19. Fondaparinux bei Herz-Kreislauf-Erkrankungen: Ein neues Antithrombin mit herausragenden Eigenschaften

    Huber K


    Full Text Available Fondaparinux, ein synthetisches Pentasaccharid, führt zu einer indirekten Hemmung des Gerinnungsfaktors Xa und behindert in der Folge die Bildung von Thrombin. Fondaparinux wurde als Vergleichssubstanz gegenüber unfraktioniertem (Standard- Heparin oder dem niedermolekularen Heparin Enoxaparin in der Prophylaxe oder Therapie von venösen Thrombosen getestet. Zuletzt wurde Fondaparinux auch bei Patienten mit akuten Koronarsyndromen (ACS untersucht: bei Patienten mit ACS ohne ST-Hebung (NSTE-ACS waren sowohl die Blutungsrate als auch die Kurz- und Langzeitmortalität im Fondaparinuxarm (2,5 mg/Tag s. c. signifikant geringer als in den Enoxaparin-behandelten Patienten (1 mg/kg KG 2×/Tag s. c. (OASIS-5-Studie. Bei Patienten mit akutem ST-Strecken-Hebungsinfarkt (STEMI war Fondaparinux in den Subgruppen der konservativ behandelten Patienten (ohne Reperfusion und der Patienten, die eine pharmakologische Reperfusion erhielten (Thrombolyse von Vorteil gegenüber Placebo oder unfraktioniertem Heparin. Hingegen zeigte sich bei Patienten mit STEMI, die einer Akut-PCI unterzogen wurden, eine starke Tendenz zugunsten von unfraktioniertem Heparin gegenüber Fondaparinux (OASIS-6-Studie. Daher wird Fondaparinux in den internationalen Richtlinien als das Antithrombin mit der günstigsten Risiko/Nutzen-Ratio bei NSTEMI aber auch bei STEMI-Patienten mit Ausnahme jener Patienten, die sich einer Akut-PCI unterziehen, empfohlen. Fondaparinux könnte schon in der nahen Zukunft die Heparine in diesen Indikationen weitgehend ersetzen.

  20. Comparative study of antithrombin III. Protease complex metabolism by fibroblasts and vascular endothelial cells

    125I-labeled human antithrombin III (125I-AT III).protease complexes are specifically bound to both cultured human skin fibroblast (HSF) cells and adult bovine aortic endothelial (ABAE) cells; however, there is a significant difference in the rate and degree of metabolism of the complexes by these two cell types. HSF cells appear to internalize the complexes at a rate of about 2.5 pmole/1 X 10(6) cells/h and subsequently degrade them at a rate of 0.6 pmole/1 X 10(6) cells/h. ABAE cells internalize and degrade the complexes at rates approximately 100 and 30 times lower, respectively. Neither cell type interacts with free 125I-AT III but only with its combined form with either thrombin or trypsin. These data indicate the major role of HSF cells in the removal of AT III.protease complexes from extravascular spaces in the body, in contrast to the inert vascular surface with regard to AT III.protease complexes provided by the vascular endothelium

  1. A capillary zone electrophoresis method to detect conformers and dimers of antithrombin in therapeutic preparations.

    Marie, Anne-Lise; Tran, Nguyet Thuy; Saller, François; Abdou, Youmna Mohamed; Zeau, Pascal; Plantier, Jean-Luc; Urbain, Rémi; Borgel, Delphine; Taverna, Myriam


    Antithrombin (AT) is a human plasma glycoprotein that possesses anticoagulant and anti-inflammatory properties. However, the native (active) form of AT is unstable and undergoes conformational changes, leading to latent, cleaved, and heterodimeric forms. The presence of these alternative forms mostly inactive can highly impact the quality and therapeutic activity of pharmaceutical AT preparations. We developed a capillary zone electrophoresis method, based on a neutral polyethylene oxide-coated capillary and a buffer close to physiological conditions, enabling the separation of more than eight forms of AT. Several peaks were identified as native, latent, and heterodimeric forms. The CZE method was reproducible with intraday relative standard deviations less than 0.5 and 2% for migration times and peak areas, respectively. The method was applied to the comparison of AT preparations produced by five competitive pharmaceutical companies, and statistical tests were performed. Important differences in the proportion of each form were highlighted. In particular, one AT preparation was shown to contain a high quantity of heterodimer, and two preparations contained high quantities of latent form. In addition, one AT preparation exhibited additional forms, not yet identified. PMID:26989842

  2. Antithrombin reduces reperfusion-induced hepatic metastasis of colon cancer cells

    Masanao Kurata; Kenji Okajima; Toru Kawamoto; Mitsuhiro Uchiba; Nobuhiro Ohkohchi


    AIM: To examine whether antithrombin (AT) could prevent hepatic ischemia/reperfusion (I/R)-induced hepatic metastasis by inhibiting tumor necrosis factor (TNF)-α-induced expression of E-selectin in rats.METHODS: Hepatic I/R was induced in rats and mice by clamping the left branches of the portal vein and the hepatic artery. Cancer cells were injected intrasplenically.The number of metastatic nodules was counted on day 7after I/R. TNF-α and E-selectin mRNA in hepatic tissue,serum fibrinogen degradation products and hepatic tissue levels of 6-keto-PGF1α, a stable metabolite of PGI2,were measured.RESULTS: AT inhibited increases in hepatic metastasis of tumor cells and hepatic tissue mRNA levels of TNF-αand E-selectin in animals subjected to hepatic I/R.Argatroban, a thrombin inhibitor, did not suppress any of these changes. Both AT and argatroban inhibited I/R-induced coagulation abnormalities. I/R-induced increases of hepatic tissue levels of 6-keto-PGF1αwere significantly enhanced by AT. Pretreatment with indomethacin completely reversed the effects of AT.Administration of OP-2507, a stable PGI2 analog, showed effects similar to those of AT in this model. Hepatic metastasis in AT-deficient mice subjected to hepatic I/R was significantly increased compared to that observed in wild-type mice. Administration of AT significantly reduced the number of hepatic metastases in congenital AT-deficient mice.CONCLUSION: AT might reduce I/R-induced hepatic metastasis of colon cancer cells by inhibiting TNF-α-induced expression of E-selectin through an increase in the endothelial production of PGI2. These findings also raise the possibility that AT might prevent hepatic metastasis of tumor cells if administered during the resection of liver tumors.

  3. Biological Variations of Lupus Anticoagulant, Antithrombin, Protein C, Protein S, and von Willebrand Factor Assays.

    Shou, Weiling; Chen, Qian; Wu, Wei; Cui, Wei


    The results of lupus anticoagulant (LA), antithrombin (AT), protein C (PC), and protein S (PS) testing, and the values of von Willebrand factor antigen (VWF:Ag) are important in diagnosis and therapeutic monitoring of thrombosis and hemostasis diseases. Till now, no published study has focused on the biological variations in LA testing, and only a few studies have examined the biological variations of AT, PC, PS, and VWF:Ag. With the latest fully automated instruments and improved reagents, the analytical, within-subject, and between-subject biological variations were estimated for these five coagulant parameters in a cohort of 25 apparently healthy subjects. Blood specimens were collected at 8:00 am, 12:00 pm, and 4:00 pm on days 1, 3, and 5. The analytical biological variation (CV(A)) values of all the parameters were less than 3%. The within-subject biological variation (CV(W)) and between-subject biological variation (CV(G)) values of the LA normalized ratio were 4.64 and 6.83%, respectively. No significant differences were observed in the intraday and interday biological variations of LA tests, or in AT, PC, PS, and VWF:Ag values. Additionally, the utility of the conventional population-based reference intervals of the five coagulation parameters was evaluated by the index of individuality, and data on CV(W) and CV(A) were used to calculate the reference change value to identify the significance of changes in serial results from the same individual. PMID:26516946

  4. Prevention and treatment of venous thromboembolism in pregnancy in patients with hereditary antithrombin deficiency

    James AH


    Full Text Available Andra H James,1 Barbara A Konkle,2,3 Kenneth A Bauer4 1Department of Obstetrics and Gynecology, University of Virginia, Charlottesville, Virginia, 2Puget Sound Blood Center, Seattle, Washington, 3Department of Medicine, University of Washington, Seattle, Washington, 4Department of Medicine, Beth Israel Deaconess Medical Center and VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA Objective: The aims of the study reported here were to provide data from six pregnant subjects who were enrolled in a clinical trial of antithrombin (AT concentrate, discuss other published case series and case reports, and provide general guidance for the use of AT concentrate for inherited AT deficiency in pregnancy. Methods: In the late 1980s, 31 AT-deficient subjects were enrolled in a prospective treatment trial of the plasma-derived AT concentrate Thrombate III®. Herein, newly available treatment data about the six pregnant subjects in the trial is tabulated and summarized. Results: All six experienced venous thromboembolism (VTE during pregnancy, were dosed according to a weight-based protocol, and were treated concomitantly with anticoagulation. Loading doses of AT concentrate of 54–62 units/kg were followed by maintenance doses of 50%–100% of the loading dose for 3–10 days. At the time of labor, loading doses of 46–50 units/kg were followed by maintenance doses of 50%–75% of the loading dose for 5–7 days. None of the six experienced recurrent thrombosis while receiving treatment with AT concentrate. Conclusion: Currently we suggest that women with AT deficiency who are pregnant or postpartum and have a personal history of VTE or current VTE receive AT concentrates. Keywords: thrombophilia, thrombosis, plasma-derived concentrate, labor, delivery, heparin.

  5. Interaction of antithrombin III with bovine aortic segments. Role of heparin in binding and enhanced anticoagulant activity.

    Stern, D.; Nawroth, P; Marcum, J; Handley, D; Kisiel, W; Rosenberg, R; Stern, K.


    Bovine antithrombin III (AT III) interaction with the luminal surface of bovine aortic segments with a continuous layer of endothelium was examined. Incubation of 125I-AT III with vessel segments, previously washed free of endogenous AT III, demonstrated specific, time-dependent binding to the protease inhibitor to the endothelium. Half-maximal binding was observed at an added AT III concentration of 14 nM. Binding of 125I-AT III to the vessel wall was reversible (50% dissociated in 4 min), a...

  6. Resolution of preoperative portal vein thrombosis after administration of antithrombin III in living donor liver transplantation: case report.

    Imai, H; Egawa, H; Kajiwara, M; Nakajima, A; Ogura, Y; Hatano, E; Ueda, M; Kawaguchi, Y; Kaido, T; Takada, Y; Uemoto, S


    A 59-year-old man with hepatitis C virus-associated liver cirrhosis was transferred to our hospital to undergo living donor liver transplantation. Coagulation was impaired (prothrombin time [International Normalized Ratio], 3.27), and antithrombin III (AT-III) activity was 23% (normal, 87%-115%). Contrast-enhanced computed tomography scans revealed portal vein thrombosis (PVT) from the junction between the splenic and superior mesenteric vein to the porta hepatica; the portal vein was completely obstructed (PVT). To prevent further development of PVT, 1500 U of AT-III was administered for 3 days, elevating the AT-III activity to 50%. A contrast-enhanced computed tomography scan obtained 9 days after AT-III administration showed resolution of PVT. Living donor liver transplantation was safely performed without portal vein grafting. Thus, a low AT-III concentration may have an important role in the pathogenesis of PVT in patients with cirrhosis. PMID:19917415

  7. Investigating changes in the gas-phase conformation of Antithrombin III upon binding of Arixtra using traveling wave ion mobility spectrometry (TWIMS)

    Zhao, Yuejie; Singh, Arunima; Li, Lingyun; Linhardt, Robert J.; Xu, Yongmei; Liu, Jian; Woods, Robert J.; Amster, I. Jonathan


    We validate the utility of ion mobility to measure protein conformational changes induced by the binding of glycosaminoglycan ligands, using the well characterized system of Antithrombin III (ATIII) and Arixtra, a pharmaceutical agent with heparin (Hp) activity. Heparin has been used as a therapeutic anticoagulant drug for several decades through its interaction with ATIII, a serine protease inhibitor that plays a central role in the blood coagulation cascade. This interaction induces conform...

  8. Development of Methods for Measuring Protein C Inhibitor and Antithrombin: Use of Monoclonal Antibodies against the Reactive Center Loop-Inserted Forms of the Serpins

    Kjellberg, Margareta


    Protein C inhibitor (PCI) and antithrombin (AT) are serine protease inhibitors (serpins) that are involved in the regulation of coagulation. Like other inhibitory serpins, PCI and AT adopt different structural conformations that are related to their functions. The cleaved, inactive form is a result of cleavage by a protease, and the latent form, which is also inactive, can arise due to a mutation in the serpin. Methods that quantify the different forms can be useful as diagnostic tools. The a...

  9. Purified radiolabeled antithrombin III metabolism in three families with hereditary AT III deficiency: application of a three-compartment model

    Purified human radioiodinated antithrombin III (125I-AT III) was used to study its metabolism in six members from three different families with a known hereditary AT III deficiency. Six healthy volunteers served as a control group. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and crossed immunoelectrophoresis (CIE) showed the purified AT III to be homogeneous. Amino acid analysis of the protein revealed a composition identical to a highly purified internal standard. The specific activity was 5.6 U/mg. Analysis of plasma radioactivity data was performed, using a three-compartment model. Neither plasma disappearance half-times nor fractional catabolic rate constants differed significantly between patients and control subjects. The mean absolute catabolic rate in the patient group was significantly lower than that of the control group at 2.57 +/- 0.44 and 4.46 +/- 0.80 mg/kg/day, respectively. In addition, the mean patient alpha 1-phase, flux ratio (k1,2 and k2,1) of the second compartment alpha 2-phase and influx (k3,1) of the third compartment were significantly reduced as compared with control values. It has been tentatively concluded that the observed reduction in the second compartment may be caused by a decrease in endothelial cell surface binding

  10. Localization of anticoagulantly active heparan sulfate proteoglycans in vascular endothelium: Antithrombin binding on cultured endothelial cells and perfused rat aorta

    We have studied the interaction of 125I-antithrombin (125I-AT) with microvascular endothelial cells (RFPEC) to localize the cellular site of anticoagulantly active heparan sulfate proteoglycans (HSPG). The radiolabeled protease inhibitor bound specifically to the above HSPG with a Kd of approximately 50 nM. Confluent monolayer RFPEC cultures exhibited a linear increase in the amount of AT bound per cell for up to 16 d, whereas suspension RFPEC cultures possessed a constant number of protease inhibitor binding sites per cell for up to 5 d. These results suggest that monolayer RFPEC cultures secrete anticoagulantly active HSPG, which then accumulate in the extracellular matrix. This hypothesis was confirmed by quantitative light and EM level autoradiography which demonstrated that the AT binding sites are predominantly located in the extracellular matrix with only small quantities of protease inhibitor complexed to the cell surface. We have also pinpointed the in vivo position of anticoagulantly active HSPG within the blood vessel wall. Rat aortas were perfused, in situ, with 125I-AT, and bound labeled protease inhibitor was localized by light and EM autoradiography. The anticoagulantly active HSPG were concentrated immediately beneath the aortic and vasa vasorum endothelium with only a very small extent of labeling noted on the luminal surface of the endothelial cells. Based upon the above data, we propose a model whereby luminal and abluminal anticoagulantly active HSPG regulate coagulation mechanism activity

  11. Circulating microparticles and the risk of thrombosis in inherited deficiencies of antithrombin, protein C and protein S.

    Campello, Elena; Spiezia, Luca; Radu, Claudia M; Bulato, Cristiana; Gavasso, Sabrina; Tormene, Daniela; Woodhams, Barry; Dalla Valle, Fabio; Simioni, Paolo


    Many subjects carrying inherited thrombophilic defects will never experience venous thromboembolism (VTE) while other individuals developed recurrent VTE with no known additional risk factors. High levels of circulating microparticles (MP) have been associated with increased risk of VTE in patients with factor V Leiden and prothrombin G20210A mutation, suggesting a possible contribution of MP in the hypercoagulability of mild genetic thrombophilia. The role of MP as additional risk factor of VTE in carriers of natural clotting inhibitors defects (severe thrombophilia) has never been assessed. Plasma levels of annexin V-MP, endothelial-derived MP (EMP), platelet-derived MP (PMP), tissue factor-bearing MP (TF+) and the MP procoagulant activity (PPL) were measured in 132 carriers of natural anticoagulant deficiencies (25 antithrombin, 63 protein C and 64 protein S defect) and in 132 age and gender-matched healthy controls. Carriers of natural anticoagulant deficiencies, overall and separately considered, presented with higher median levels of annexin V-MP, EMP, PMP, TF+MP and PPL activity than healthy controls (pEMP and PMP had an adjusted OR for VTE of 3.36 (95% CI, 1.59 to 7.11), 9.26 (95% CI, 3.55 to 24.1) and 2.72 (95%CI, 1.16 to 6.38), respectively. Elevated levels of circulating MP can play a role in carriers of mild and severe inherited thrombophilia. The clinical implications of this association remain to be defined. PMID:26354831

  12. Impact of antithrombin Ⅲ on hepatic and intestinal microcirculation in experimental liver cirrhosis and bowel inflammation: An in vivo analysis

    Sasa-Marcel Maksan; Zilfi (U)lger; Martha Maria Gebhard; Jan Schmidt


    AIM: To analyze the hepatic and intestinal microcirculation in an animal model of liver cirrhosis and inflammatory bowel disease (IBD) and to characterize the anti-inflammatory action of antithrombin Ⅲ (ATⅢ) on leukocyte kinetics and liver damage.METHODS: Hepatic and intestinal microcirculation was investigated by intravital videomicroscopy. Standardized models of experimental chronic liver cirrhosis and bowel inflammation were employed. Animals were divided into four groups (n = 6/group): controls, animals with cirrhosis,animals with cirrhosis and IBD, animals with cirrhosis and IBD treated with ATⅢ.RESULTS: Cirrhosis facilitated leukocyte rolling and sticking in hepatic sinusoids (1.91±0.28 sticker/μm vs0.5±0.5 sticker/μm in controls, P<0.05). The effect enhanced in animals with cirrhosis and IBD (5.4±1.65sticker/μm), but reversed agter ATⅢ application (3.97±1.04sticker/μm, P<0.05). Mucosal blood flow showed no differences in cirrhotic animals and controls (5.3±0.31nL/min vs5.4±0.25 nL/min) and was attenuated in animals with cirrhosis and IBD significantly (3.49±0.6 nL/min). This effect was normalized in the treatment group (5.13±0.4nL/min, P<0.05). Enzyme values rose during development of cirrhosis and bowel inflammation, and reduced after ATⅢ application (P<0.05).CONCLUSION: Liver cirrhosis in the presence of IBD leads to a significant reduction in mucosal blood flow and an increase in hepatic leukocyte adherence with consecutive liver injury, which can be prevented by administration of ATⅢ.

  13. Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides.

    Yamada, Tsuyoshi; Kanda, Yutaka; Takayama, Makoto; Hashimoto, Akitoshi; Sugihara, Tsutomu; Satoh-Kubota, Ai; Suzuki-Takanami, Eri; Yano, Keiichi; Iida, Shigeru; Satoh, Mitsuo


    The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. In human plasma, AT circulates as a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135. However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. Here, we succeeded in directly comparing the activities between the high-mannose and complex types. Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. PMID:26747427

  14. A prospective cohort study on the absolute risks of venous thromboembolism and predictive value of screening asymptomatic relatives of patients with hereditary deficiencies of protein S, protein C or antithrombin.

    Mahmoodi, B.K.; Brouwer, J.L.P.; Kate, M.K. Ten; Lijfering, W.M.; Veeger, N.J.; Mulder, A.B.; Kluin-Nelemans, H.C.; Meer, J. van der


    BACKGROUND: Absolute risks of venous thromboembolism (VTE) in protein S-, protein C-, or antithrombin-deficient subjects are mainly based on retrospective data. Screening asymptomatic relatives of these patients is disputed, though studies addressing this issue have yet to be conducted. METHODS: We

  15. An antithrombin-dependent sulfated polysaccharide isolated from the green alga Caulerpa cupressoides has in vivo anti- and prothrombotic effects Um polissacarídeo sulfatado dependente de antitrombina isolado da alga verde Caulerpa cupressoides possui efeitos anti- e pró-trombótico in vivo

    José Ariévilo Gurgel Rodrigues; Ismael Nilo Lino de Queiroz; Ana Luíza Gomes Quinderé; Bruno Cunha Vairo; Paulo Antônio de Souza Mourão; Norma Maria Barros Benevides


    Red algae sulfated polysaccharides (SPs) have been widely described as anticoagulant and antithrombotic agents; however no description of antithrombotic activity regarding green algae SPs has been reported. Caulerpa cupressoides (Chlorophyta) has three different SPs fractions (SP1, SP2 and SP3). We investigated the effects of SP2 on thrombin activity by antithrombin and in an experimental model of venous thrombosis in rats. The inhibition of thrombin assay was evaluated using antithrombin (AT...

  16. PEGylation of Hirudin and Analysis of Its Antithrombin Activity in vitro%水蛭素聚乙二醇化及其体外抗凝活力分析

    秦海娜; 修志龙; 张代佳; 包永明; 李晓晖; 韩国柱


    Hirudin is the most anticoagulant drug found in nature, but its short serum half-life significantly inhibits its clinical application. The PEGylation of hirudin, the most promising anticoagulant drug, was performed in this paper. The optimal reaction conditions for PEGylated hirudin were investigated. When the PEGylation reaction was conducted under 4℃ after 10h, in the borate buffer at pH 8.5, with the molar ratio 250: 1 of PEG to hirudin, a higher modification extent was achieved. Finally, the bioactivity of PEGylated hirudin was measured in vitro.Compared with unmodified hirudin, 26% of anti-thrombin activity was retained.

  17. Investigating changes in the gas-phase conformation of Antithrombin III upon binding of Arixtra using traveling wave ion mobility spectrometry (TWIMS).

    Zhao, Yuejie; Singh, Arunima; Li, Lingyun; Linhardt, Robert J; Xu, Yongmei; Liu, Jian; Woods, Robert J; Amster, I Jonathan


    We validate the utility of ion mobility to measure protein conformational changes induced by the binding of glycosaminoglycan ligands, using the well characterized system of Antithrombin III (ATIII) and Arixtra, a pharmaceutical agent with heparin (Hp) activity. Heparin has been used as a therapeutic anticoagulant drug for several decades through its interaction with ATIII, a serine protease inhibitor that plays a central role in the blood coagulation cascade. This interaction induces conformational changes within ATIII that dramatically enhance the ATIII-mediated inhibition rate. Arixtra is the smallest synthetic Hp containing the specific pentasaccharide sequence required to bind with ATIII. Here we report the first travelling wave ion mobility mass spectrometry (TWIMS) investigation of the conformational changes in ATIII induced by its interaction with Arixtra. Native electrospray ionization mass spectrometry allowed the gentle transfer of the native topology of ATIII and ATIII-Arixtra complex. IM measurements of ATIII and ATIII-Arixtra complex showed a single structure, with well-defined collisional cross section (CCS) values. An average 3.6% increase in CCS of ATIII occurred as a result of its interaction with Arixtra, which agrees closely with the theoretical estimation of the change in CCS based on protein crystal structures. A comparison of the binding behavior of ATIII under both denaturing and non-denaturing conditions confirmed the significance of a folded tertiary structure of ATIII for its biological activity. A Hp oligosaccharide whose structure is similar to Arixtra but missing the 3-O sulfo group on the central glucosamine residue showed a dramatic decrease in binding affinity towards ATIII, but no change in the mobility behavior of the complex, consistent with prior studies that suggested that 3-O sulfation affects the equilibrium constant for binding to ATIII, but not the mode of interaction. In contrast, nonspecific binding by a Hp

  18. Antithrombin III (AT and recombinant tissue plasminogen activator (R-TPA used singly and in combination versus supportive care for treatment of endotoxin-induced disseminated intravascular coagulation (DIC in the neonatal pig

    Gardner Renee'


    Full Text Available Abstract Background Disseminated intravascular coagulation (DIC is a pathological disturbance of the complex balance between coagulation and anticoagulation that is precipitated by vascular injury, acidosis, endotoxin release and/or sepsis and characterized by severe bleeding and excessive clotting. The innately low levels of coagulation factors found in newborn infants place them at extremely high risk for DIC. Anecdotal reports suggest that either anticoagulant or fibrinolytic therapy may alleviate some of the manifestations of DIC. To test the hypothesis that replacement of both anticoagulants and fibrinolytics may improve survival and outcome better than either single agent or supportive care alone, we utilized a neonatal piglet model of endotoxin-induced DIC. Methods DIC was induced in twenty-seven neonatal pigs (7 to 14 days of age by intravenous administration of E. coli endotoxin (800 μg/kg over 30 min. The piglets were divided into 4 groups on the basis of treatment protocol [A: supportive care alone; B: Antithrombin III (AT, 50 μg/kg bolus, 25 μg/kg per hr continuous infusion and supportive care; C: Recombinant Tissue Plasminogen Activator (R-TPA, 25 μg/kg per hr continuous infusion and supportive care; D: AT, R-TPA and supportive care] and monitored for 3 primary outcome parameters (survival time, macroscopic and microscopic organ involvement and 4 secondary outcome parameters (hematocrit; platelet count; fibrinogen level; and antithrombin III level. Results Compared with supportive care alone, combination therapy with AT and R-TPA resulted in a significant improvement of survival time, hematocrit, AT level, macroscopic and microscopic organ involvement, p Conclusion The findings suggest that combining AT, R-TPA and supportive care may prove more advantageous in treating the clinical manifestations of DIC in this neonatal pig model than either single modality or supportive care alone.

  19. Correlative Analysis of the Relationships and Differences on D-dimer, Fibrinogen and Antithrombin Ⅲ Between Varied Clinical Periods of Cerebral Infarction%D-D、Fb和AT-Ⅲ在脑梗死不同时期的变化及相关性分析

    徐威香; 武蓉珍; 胡晓蕾


    目的:探讨血浆D-二聚体(D-D)、纤维蛋白原(Fb)和抗凝血酶Ⅲ(AT-Ⅲ)在脑梗死(CI)不同时期的变化及其临床意义.方法:根据临床特征,对260例CI患者进行分组,其中CI急性期组80例、进展期组41例、非进展期组65例、康复期组74例;90例体检健康者为正常对照组.采用SYSMEX CA7000血凝仪分别检测其血浆D-D、Fb和AT-Ⅲ水平.结果:CI急性期、进展期、非进展期患者D-D、Fb含量均明显高于正常对照组(P<0.01),AT-Ⅲ低于正常对照组(P<0.01);脑梗死组中,D-D与AT-Ⅲ呈负相关(P<0.01),D-D与Fb呈正相关(P<0.01),AT-Ⅲ与Fb无相关性.结论:CI患者D-D、Fb、AT-Ⅲ变化显著,D-D升高伴随AT-Ⅲ含量降低,提示D-D、Fb、AT-Ⅲ共同参与了梗死发生发展的病理生理过程,可作为CI临床危险分级和病情监测的指标.%Objective To investigate the differences and its clinical significance of D-dimer, fibrinogen and antithrombin Ⅲ between varied clinical periods of cerebral infarction( CI). Methods 328 CI patients were rolled into the study, and they were divided into four groups due to their different clinical periods as follows: group acute period (80) , group progressing(41) , group non-pro-gressing(65) , group convalescence(74). And, 90 healthy volunteers were rolled into the normal control group. Their plasma D-dimer, fibrinogen and antithrombin HI were detected on SYSMEX CA7000 automatic coagulation analyzer. Results D-dimer and fibrinogen of CI patients in the acute, progressing and non-progressing periods were significantly higher (P <0.01) than those of healthy volunteers, while antithrombin Ⅲ of them was significantly lower (P<0.01) than that of healthy volunteers. In CI patients, D-dimer correlated negatively with antithrombin Ⅲ, while positively with fibrinogen. No significant relationship was observed between antithrombin Ⅲ and fibrinogen. ConclUSin D-dimer, fibrinogen and antithrombin Ⅲ of CI patients varied significantly

  20. Interstitial deletion of chromosome 1q [del(1)(q24q25.3)] identified by fluorescence in situ hybridization and gene dosage analysis of apolipoprotein A-II, coagulation factor V, and antithrombin III

    Takano, Takako; Yamanouchi, Yasuko; Mori, Yosuke [Teikyo Univ. School of Medicine, Tokyo (Japan)] [and others


    We report on a 12-month-old Japanese boy with an interstitial deletion of the long-arm of chromosome 1 and meningomyelocele, hydrocephalus, anal atresia, atrial septal defect, left renal agenesis, bilateral cryptorchidism, talipes equinovarus, low birth weight, growth/developmental retardation, and many minor anomalies. By conventional GTG-banding, his karyotype was first interpreted as 46,XY,de1(1)(q23q24), but it was corrected as 46,XY.ish del(1)(q24q25.3) by fluorescence in situ hybridization using 11 known cosmid clones as probes. His serum levels of apolipoprotein A-II (gene symbol: APOA2, previously assigned to 1q21-q23) and coagulation factor V (F5, 1q21-q25) were normal, while serum concentration and activity of antithrombin III (AT3, 1q23-q25.1) was low. The results indicated that localization of APOA2 and F5 are proximal to the deleted region and AT3 is located within the deletion extent in the patient. 16 refs., 4 figs.

  1. The functional study of antithrombin L99 mutation%抗凝血酶L99氨基酸位点突变对其功能的影响

    郁婷婷; 戴菁; 丁秋兰; 傅启华; 王学锋


    Objective To study the molecular mechanisms of inherited antithrombin (AT)defiency caused by AT L99 mutation.Methods Wild type (WT),L99V,L99A,L99I and L99S AT were purified from drosophila expression system.The binding capacity of AT and the low molecular weight heparin sodium was analyzed by the heparin binding assay.Surface plasmon resonance (SPR) was used to detect the binding ability of AT to thrombin (F Ⅱ a) or AT to coagulation factor X a (F X a).The activity of AT (AT∶ A) was detected by chromogenic assay.Results The purified WT and mutant AT were at the same size.No additional band was observed by coomassie blue staining and western blot assay.Compared to the WT AT,the binding abilities of the low molecular weight heparin sodium to the AT L99V,L99A,L99I and L99S were (44.8±3.6)%,(118.9±14.0)%,(15.2±8.8)%,and (23.0±8.2)%,respectively.The binding abilities of F Ⅱ a to AT L99V,L99A,L99I and L99S were 13%,57%,3%,and 29%,while the binding of F X a to AT L99V,L99A,L99I and L99S were 7%,51%,1%,and 25%.The AT∶A of WT,L99V,L99A,L99I and L99S AT were 146.5%,21.4%,120.9%,10.8%,and 39.0%,respectively.Conclusion The binding abilities of AT to heparin,F Ⅱ a and F X a were damaged by the L99 mutation,which resulted in decreased AT∶ A and inherited AT deficiency.%目的 研究抗凝血酶(AT)L99氨基酸位点突变导致遗传性AT缺陷症的分子机制.方法 利用果蝇细胞表达系统表达纯化野生型AT以及L99V、L99A、L99I和L99S等突变型AT蛋白;采用肝素结合实验检测重组AT蛋白与低分子量肝素钠的结合能力;采用表面等离子共振(SPR)技术检测重组AT蛋白与凝血酶(FⅡa)及活化凝血因子X(FXa)的结合能力;将野生型及各突变型AT蛋白的浓度调整至正常AT血浆浓度后,采用发色底物法检测重组AT蛋白活性.结果 考马斯亮蓝染色及免疫印迹法显示各突变蛋白与野生型AT蛋白大小一致,未见明显杂带.

  2. An antithrombin-dependent sulfated polysaccharide isolated from the green alga Caulerpa cupressoides has in vivo anti- and prothrombotic effects Um polissacarídeo sulfatado dependente de antitrombina isolado da alga verde Caulerpa cupressoides possui efeitos anti- e pró-trombótico in vivo

    José Ariévilo Gurgel Rodrigues


    Full Text Available Red algae sulfated polysaccharides (SPs have been widely described as anticoagulant and antithrombotic agents; however no description of antithrombotic activity regarding green algae SPs has been reported. Caulerpa cupressoides (Chlorophyta has three different SPs fractions (SP1, SP2 and SP3. We investigated the effects of SP2 on thrombin activity by antithrombin and in an experimental model of venous thrombosis in rats. The inhibition of thrombin assay was evaluated using antithrombin (AT in the presence of SP2 and the antithrombotic activity was investigated in rats with thromboplastin as the thrombogenic stimulus. The anticoagulant effects of SP2 are suggested be due to the potentiation of thrombin inhibition by antithrombin (IC50 ~ 10.0µg mL-1 and this mechanism of interaction is different when compared to other studied Caulerpa polysaccharides. SP2 exhibited antithrombotic effects at doses of 1.0 and 2.0mg kg-1 body weight, but at higher doses (>2.0mg kg-1 body weight this polysaccharide revert the antithrombotic property. No hemorrhagic effect (2.0mg kg-1 was observed. As occurs with red algae SPs, these results indicate that green algae SPs are also capable of exhibiting different in vivo properties.Os polissacarídeos sulfatados (PSs de algas vermelhas têm sido relatados mundialmente como agentes anticoagulantes e antitrombóticos. Entretanto, nenhuma descrição de atividade antitrombótica tem sido relacionada com os PSs de algas verdes. A clorofícea Caulerpa cupressoides possui três frações de PSs (PS1; PS2 e PS3. Dessa forma, objetivou-se investigar os efeitos da fração PS2 sobre a atividade da trombina por antitrombina e usando um modelo experimental de trombose venosa em ratos. O ensaio de inibição da trombina foi avaliado usando a antitrombina (AT na presença de PS2 e a atividade antitrombótica foi investigada em ratos, usando a tromboplastina como o estímulo trombogênico. Os efeitos anticoagulantes de PS2 devem

  3. Deficiencies of Natural Anticoagulants, Protein C, Protein S, and Antithrombin

    ... citing article information Citing articles via Google Scholar Google Scholar Articles by Lipe, B. Articles by Ornstein, D. L. Search for related content PubMed PubMed citation Articles by Lipe, B. Articles by Ornstein, D. ... Reddit StumbleUpon Twitter What's this? Navigate This ...

  4. Clinical significance on changes of platelet aggregation test, von Willebrand factor,antithrombin and D-dimer assayin acute cerebral infarction patients%急性脑梗死患者血小板聚集功能、血管性血友病因子、抗凝血酶及 D-二聚体测定的临床意义

    叶青跃; 程鹏飞; 周有利; 饶汉武; 黄承芳; 周立


    目的:探讨急性脑梗死患者血小板聚集功能( PAgT)、血管性血友病因子( vWF)、抗凝血酶( AT)和D-二聚体( D-dimer)水平变化及临床意义。方法选用相应的方法和仪器测定112例脑梗死及80例健康对照者血(浆) PAgT、vWF、AT和D-dimer水平变化,同时对部分患者进行治疗前、后的对比分析。结果脑梗死患者血中PAgT、vWF、D-dimer等指标均明显高于健康对照组,AT活性较对照组显著降低,差异有统计学意义(P<0.05或P<0.01)。选取经治疗效果明显好转的78例脑梗死患者,出院前取空腹静脉血测定PAgT、vWF、AT、D-dimer等指标,并与治疗前对照,结果治疗后PAgT、vWF、D-dimer降低,AT活性升高,差异有统计学意义(P<0.05或P<0.01)。结论脑梗死患者体内存在明显的凝血及纤溶功能异常,与血管内皮损伤、血小板聚集功能增强、凝血及纤溶功能亢进、抗凝功能降低等多因素有关。 PAgT、vWF、AT、D-dimer可以作为脑梗死患者诊断、治疗监测和预后判断的参考指标。%Objective To evaluate the clinical signification of coagulation ,anti-coagulation and fibrinolysis indexes i.e.platelet aggrega-tion test(PAgT),von Willebrand factor(vWF),antithrombin(AT),D-dimer in acute cerebral infarction patients.Methods vWF was as-sayed using ELISA method,AT was determined by chromogenic substances assay,and Latex enhanced immune turbidimetry for D-dimer. vWF,AT and D-dimer all the parameters were finished by SysmexCA-7000 automated blood coagulation analyzer.PAgT was measured sim-ultaneously using a whole-blood Lumi-Aggregometer by CHRMNO-LOG platelet aggregation apparatus.Results PAgT, vWF, D-dimer were significantly higher in acute cerebral infarction patients group,compared with those in the control group(P<0.05 or P<0.01). while AT was significantly lower(P<0.05).After effective treatment,PAgT,vWF,AT,D-dimer Indicators are all

  5. Correlation of the activities of protein C,antithrombin and coagulation factor Ⅷ with the treatment of lung cancer with pulmonary embolism%蛋白 C、抗凝血酶及凝血因子Ⅷ活性变化与肺癌合并肺栓塞后治疗的关系

    陈环; 张鹏; 刘军锋


    Objective To investigate the correlation of the activities of protein C(PC),antithrombin(AT)and coagulation factor Ⅷ(FⅧ)with the treatment of lung cancer with pulmonary embolism(PE).Methods A total of 98 patients with lung cancer and PE were enrolled as lung cancer with PE group.A total of 1 00 patients with lung cancer were enrolled as lung cancer group.Their sex and age were recorded,and lipoprotein(a)[Lp(a)],total cholesterol (TC),triglyceride (TG),C-reactive protein(CRP),thrombin time(TT),platelet (PLT)and fibrinogen(Fbg)in the 2 groups were determined,respectively.PC,AT,FⅧ and D-dimer were determined in lung cancer with PE group after 5-7 d of treatment.Logistic regression analysis was used to analyze the factors influencing coagulation-fibrinolysis after 5-7 d of treatment in lung cancer with PE group.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic efficiency of these factors.Results Lp (a),TC,TG and TT in lung cancer with PE group were significantly higher than those in lung cancer group (P 0.90.Conclusions The influence of PC,AT and FⅧ on coagulation-fibrinolysis should be concerned in midterm treatment of lung cancer with PE,and their activities can be used to assess the therapeutic efficiency and treatment program in a given period.%目的:探讨蛋白C(PC)、抗凝血酶(AT)及凝血因子Ⅷ(FⅧ)活性变化与肺癌合并肺血栓栓塞[简称肺栓塞(PE)]后治疗的关系。方法选择肺癌合并 PE 患者(肺癌+PE 组)98例及肺癌患者(肺癌组)100例。记录两组性别、年龄并检测脂蛋白(a)[Lp(a)]、总胆固醇(TC)、甘油三酯(TG)、C 反应蛋白(CRP)、凝血酶时间(TT)、血小板数量(PLT)、纤维蛋白原(Fbg)等指标。检测肺癌+PE 组治疗5~7 d 及肺癌组入院时的 PC、AT、FⅧ、D-二聚体水平。应用 Logistic 回归分析肺癌合并 PE 后治疗5~7 d 凝血-纤溶状态的影响因素,

  6. Clinical Significance of von Willebrand Factor and Antithrombin in Diabetes Mellitus with Retinopathy%血管性血友病因子与抗凝血酶在糖尿病视网膜病变中的临床意义

    张鹏; 汤荣华


    目的 探讨糖尿病微血管视网膜病变患者血管性血友病因子(vWF)、抗凝血酶(AT)的变化及临床意义.方法 将100例2型糖尿病患者分为两组,糖尿病无微血管视网膜病变并发症组53例,糖尿病微血管视网膜病变并发症组47例,设健康人群对照组50例.所有人群入院或体检时即采集静脉血,进行vWF:Ag、AT:A检测.结果 2型糖尿病无微血管视网膜病变并发症组患者血浆vWF:Ag含量较正常对照组增高(P<0.05),2型糖尿病微血管视网膜病变并发症组患者血浆vWF:Ag含量与2型糖尿病无微血管视网膜病变并发症组及正常对照组相比有显著性差异(P<0.05);且2型糖尿病微血管视网膜病变并发症组中背景性病变和增殖性病变患者血浆vWF:Ag含量比较有显著性差异(P<0.05).2型糖尿病无微血管视网膜病变并发症组患者血浆AT活性与正常对照组相比无显著性差异(P>0.05),2型糖尿病微血管视网膜病变并发症组患者血浆AT活性与2型糖尿病无微血管视网膜病变并发症组及正常对照组相比有显著性差异(P<0.05);且2型糖尿病微血管视网膜病变并发症组中背景性病变和增殖性病变患者血浆AT活性比较有显著性差异(P<0.05).结论 血浆vWF:Ag、AT:A的变化与糖尿病微血管视网膜病变的发生、发展有密切关系,监测其水平与活性对糖尿病微血管并发症的治疗和预防有重要的临床意义.%Objective To explore the changes of von Willebrand factor(vWF) and antithrombin(AT) in diabetic patients with retinopathy and its clinical implications. Methods 100 type 2 diabetes mellitus (T2DM) patients were divided into 2 groups,one group included 53 T2DM patients without retinopathy,another group included 47 T2DM patients with retinopathy, 50 healthy peoples were enrolled as control group. Venous blood was collected from all groups and vWF:Ag, AT; A were determined. Results The level of vWF;Ag in T2DM

  7. Developmental expression of chicken antithrombin III is regulated by increased RNA abundance and intracellular processing.

    Amrani, D L; Rosenberg, J; Samad, F; Bergtrom, G; Banfield, D K


    We isolated and sequenced a 432 bp cDNA to cAT-III, that encoded 115 nucleotides of 5' untranslated sequence, a 17 amino acid long signal peptide and residues 1-88 of the mature protein, and used it to prepare a probe for measuring and correlating the developmental changes of steady-state cAT-III mRNA levels with known changes in antigen levels. Densitometric analysis of nuclease protection (n = 2), Northern blot (n = 4), and slot blots (n = 3) of total RNA from chick livers of 16-day-old embryos to 6-day-old chicks showed a 2.6 +/- 0.5-fold increase in steady-state cAT-III mRNA levels. Assay of functional mRNA levels by in vitro translation of poly(A)+ RNA and specific immunoprecipitation of 35S-Met-labelled cAT-III was comparable to RNA analysis (16-day-old embryos vs. 10-day-old hatchlings). We evaluated whether there were developmental differences in post-translational secretion which may also contribute to the regulation of the circulating level of this protein. Pulse-chase studies of freshly-isolated hepatocytes from 16-day-old embryos and 10-day-old hatchlings maintained in suspension demonstrated a approx. 5.0-5.5-fold increase in cAT-III levels at steady-state secretion. The above findings indicate that changes in circulating cAT-III levels during late embryonic development are primarily due to increased abundance of cAT-III mRNA. In addition, we postulate that post-translational intracellular processing may account for further differences in circulating protein levels. PMID:8424948

  8. Prevention and treatment of venous thromboembolism in pregnancy in patients with hereditary antithrombin deficiency

    James AH; Konkle BA; Bauer KA


    Andra H James,1 Barbara A Konkle,2,3 Kenneth A Bauer4 1Department of Obstetrics and Gynecology, University of Virginia, Charlottesville, Virginia, 2Puget Sound Blood Center, Seattle, Washington, 3Department of Medicine, University of Washington, Seattle, Washington, 4Department of Medicine, Beth Israel Deaconess Medical Center and VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA Objective: The aims of the study reported here were to provide data from six pregnan...

  9. Anti-thrombin therapy during warm ischemia and cold preservation prevents chronic kidney graft fibrosis in a DCD model

    Favreau, F.; Thuillier, R.; Cau, J; S. Milin; Manguy, E; Mauco, G; Zhu, X.; Lerman, LO; Hauet, T.


    Ischemia reperfusion injury (IRI) is pivotal for renal fibrosis development via peritubular capillaries injury. Coagulation represents a key mechanism involved in this process. Melagatran® (M), a thrombin inhibitor, was evaluated in an autotransplanted kidney model, using Large White pigs. To mimic deceased after cardiac death donor conditions, kidneys underwent warm ischemia (WI) for 60 min before cold preservation for 24 hours in University of Wisconsin solution. Treatment with M before WI ...

  10. Antitrombina: 25 años después: Un tributo a una permanente fuente de conocimiento en Hemostasia Antithrombin: 25 years later: A tribute to a long lasting source of knowledge in Hemostasis

    Jorge Korin


    En los últimos 25 años se ha revolucionado el conocimiento de la fisiología de la Hemostasia con un rápido traslado de muchos de los nuevos hallazgos a la fisiopatología de los estados trombofílicos. El presente artículo condensa algunos de dichos avances en torno a las serpinas en general y a la antitrombina en especial. Se analiza el papel de la principal inhibidora de trombina en la hemostasia normal y en patologías trombóticas congénitas y adquiridas como así también la importante contrib...

  11. Deep venous thrombosis

    ... Blood clot in the legs; Thromboembolism; Post-phlebitic syndrome; Post-thrombotic syndrome ... for the Factor V Leiden mutation) Antithrombin levels Antiphospholipid antibodies Complete blood count (CBC) Genetic testing to ...

  12. Effect of estrogen-progestin hormonal replacement therapy on blood coagulation and fibrinolysis in postmenopausal women Efeitos da terapia de reposição hormonal estroprogestativa sobre o sistema de coagulação e de fibrinólise em mulheres na pós-menopausa

    Claudio E Bonduki; Dayse M Lourenço; Eduardo L.A. da Motta; José Maria Soares Jr; Mauro Abi Haidar; Baracat, Edmund C


    OBJECTIVE: To evaluate antithrombin III (AT), thrombin (Fragment 1+2 [F1+2] and thrombin-antithrombin [TAT]) generation markers, as well as other coagulation parameters, such as prothrombin time, partial activated thromboplastin time, thrombin time, fibrinogen, euglobulin lysis time, and platelet count, in postmenopausal women after hormonal therapy. STUDY DESIGN: Forty-five patients who received either 0.625 mg/day unopposed oral conjugated equine estrogen (CEE), 0.625 mg/day oral CEE plus m...

  13. 非ST段抬高急性冠脉综合征患者随机应用依诺肝素或普通肝素抗凝治疗的疗效和出血并发症系统综述%Efficacy and Bleeding Complications Among Patients Randomized to Enoxaparin or Unfractionated Heparin for Antithrombin Therapy in Non-ST-Segment Elevation Acute Coronary Syndromes A Systematic Overview

    John L. Petersen; James A. de Lemos; Christopher C. Nessel; Robert A. Harrington; James J. Ferguson; Eugene Braunwald; Robert M. Califf; 徐成斌; Kenneth W. Mahaffey; Vic Hasselblad; Elliott M. Antman; Marc Cohen; Shaun G. Goodman; Anatoly Langer; Michael A. Blazing; Anne Le-Moigne-Amrani


    背景:抗凝疗法已成为急性冠脉综合征(acute coronary syndrome,ACS)治疗指南推荐的标准疗法.但是,最近某些试验在ACS患者中对依诺肝素(enoxaparin)与普通肝素(unfractionated heparin)的应用进行了比较,发现这些抗凝疗法的疗效及安全性并不及既往试验结果.目的:有6项随机对照试验对依诺肝素与普通肝素治疗ACS患者进行了比较,对其终点(即全因死亡及非致死性心肌梗死)、输血与大出血进行系统评估.资料来源:从杜克临床研究所(Duke Clinical Research Institute)获取ESSENCE、A to Z及SYNERGY试验的原始数据.由TIMI 11B、ACUTEⅡ及INTERACT研究的主要研究人员提供各自的基线特征和事件发生频率.研究选取:在非ST段抬高ACS患者中比较依诺肝素与普通肝素的6项随机对照试验均入选进行分析.数据提取:从全部试验人群和随机分组前未接受抗凝治疗的亚人群中获取疗效终点和安全性终点.数据综合:应用随机效应经验性贝叶斯模型(random-effects empirical Bayes model),系统评估21 946例患者的结果.依诺肝素与普通肝素30天死亡率无显著差异(3.0%比3.0%,优势比[odds ratio,OR],1.00;95%可信区间[confidence interval,CI],0.85~1.17).在所有试验人群中,依诺肝素与普通肝素相比,30天死亡或非致死性心肌梗死(myocardial infarction,MI)联合终点显著下降,具有统计学差异(10.1%比11.0%;OR,0.91;95%CI,0.83~0.99;所需治疗例数107).随机分组前未接受抗凝治疗的依诺肝素组患者30天死亡或MI联合终点亦显著下降,具有统计学差异(8.0%比9.4%;OR,0.81;95%CI,0.70~0.94;所需治疗例数72).随机分组后第7天,总体安全人群或者随机分组前未接受抗凝治疗的人群输血(OR,1.01;95%CI,0.89~1.14)或大出血的发生率(OR,1.04;95%CI,0.83~1.30)无显著差异.结论:对近22 000例各类ACS患者进行系统回顾发现,在预防死亡或MI联合终点方面依诺肝素较普通肝素更有效.

  14. The fabrication, characterization and application of aptamer-functionalized Si-nanowire FET biosensors

    Kim, Ki Su; Lee, Hyun-Seung; Yang, Jeong-A; Jo, Moon-Ho; Hahn, Sei Kwang [Department of Materials Science and Engineering, Pohang University of Science and Technology (POSTECH), San 31, Hyoja-dong, Nam-gu, Pohang, Kyungbuk 790-784 (Korea, Republic of)], E-mail:


    An aptamer-functionalized silicon-nanowire (Si-NW) field effect transistor (FET) biosensor was successfully fabricated, characterized and applied to real-time electrical detection of binding with the target protein for biomedical applications. Surface modifications were carried out using 3-aminopropyl diethoxysilane and succinic anhydride to introduce amine and carboxyl groups onto Si substrates. Anti-thrombin aptamers with 5{sup '}-end amine groups were chemically grafted onto the surface-modified Si substrates through amide bond formation. Atomic force microscopic (AFM) analyses confirmed the successful immobilization of anti-thrombin aptamers on Si-NWs and their binding with thrombin samples. The anti-thrombin aptamers bound to Si-NWs through the linker appeared to have a mean height of approx. 4 nm and the thrombin/aptamer complex to have a mean height of approx. 8 nm. Fluorescence micrographs visualized the FITC-labeled thrombin after binding to anti-thrombin aptamers immobilized on Si-NWs. Furthermore, the anti-thrombin Si-NW FET biosensor was successfully applied to the real-time detection of electronic signals during and after binding with a thrombin sample at a concentration of approx. 330 pmol l{sup -1} and the thrombin in blood samples.

  15. Combination of a SAW-biosensor with MALDI mass spectrometric analysis.

    Treitz, G; Gronewold, T M A; Quandt, E; Zabe-Kühn, M


    A S-sens K5 surface acoustic wave biosensor was coupled with mass spectrometry (SAW-MS) for the analysis of a protein complex consisting of human blood clotting cascade factor alpha-thrombin and human antithrombin III, a specific blood plasma inhibitor of thrombin. Specific binding of antithrombin III to thrombin was recorded as a function of time with a S-sens K5 biosensor. Two out of five elements of the sensor chip were used as references. To the remaining three elements coated with RNA anti-thrombin aptamers, thrombin and antithrombin III were bound consecutively. The biosensor measures mass changes on the chip surface showing that 20% of about 400fmol/cm2 thrombin formed a complex with the 1.7-times larger antithrombin III. Mass spectrometry (MS) was applied to identify the bound proteins. Sensor chips with aptamer-captured (1) thrombin and (2) thrombin-antithrombin III complex (TAT-complex) were digested with proteases on the sensor element and subsequently identified by peptide mass fingerprint (PMF) with matrix assisted laser desorption/ionization time-of-flight (MALDI-ToF) mass spectrometry. A significant identification of thrombin was achieved by measuring the entire digest with MALDI-ToF MS directly from the sensor chip surface. For the significant identification of both proteins in the TAT-complex, the proteolytic peptides had to be separated by nano-capillary-HPLC prior to MALDI-ToF MS. SAW-MS is applicable to protein interaction analysis as in functional proteomics and to miniaturized diagnostics. PMID:18316185

  16. Dicty_cDB: Contig-U05497-1 [Dicty_cDB

    Full Text Available 4 8.4 1 ( DJ452293 ) Method for analysing genome using microsatellite ... 44 8.4 1 ( DJ047936 ) CD10-specific antibody composition.... 44 8.4 1 ( DI155118 ) Process for producing Antithrombin III composition. 44 8.4 1 (...( DD152135 ) Fusion protein composition. 44 8.4 1 ( DD152031 ) Method of producing Antithrombin III composition.... 44 8.4 1 ( DD152009 ) IL-5 receptor-specific antibody composition. 44 8.4 1 ( DD151990 ) CCR4-specific antibody composition...sition. 44 8.4 1 ( DD151869 ) Ganglioside GD3-specific antibody composition. 44 8.4

  17. The Anopheles gambiae cE5, a tight- and fast-binding thrombin inhibitor with post-transcriptionally regulated salivary-restricted expression

    Ronca, R.; Kotsyfakis, Michalis; Lombardo, F.; Rizzo, C.; Currà, C.; Ponzi, M.; Fiorentino, G.; Ribeiro, J.M.C.; Arcà, B.


    Roč. 42, č. 9 (2012), s. 610-620. ISSN 0965-1748 R&D Projects: GA ČR GAP502/12/2409 Institutional research plan: CEZ:AV0Z60220518 Keywords : Anopheles * Salivary protein * Anti-thrombin * Anophelin * Hematophagy * Post-transcriptional regulation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.234, year: 2012

  18. Kombineret antitrombinmangel og faktor VLeiden-mutation i en dansk familie

    Nielsen, Christiane; Gram, Jørgen

    Factor VLeiden (FVL) and antithrombin (AT) deficiency are both known genetic risk factors for venous thromboembolism (VTE). The combination is rare and leads to significantly increased risk for VTE. In this case we present a 22-year-old woman with VTE while treated with oral contraceptives. Labor...

  19. Drug: D08795 [KEGG MEDICUS

    Full Text Available BR:br08301] 6 Agents against pathologic organisms and parasites 63 Biological preparation...s 634 Human blood preparations 6343 Plasma preparations D08795 Human anti-thrombin III, freeze-dried concentrated PubChem: 96025478 ...

  20. A rare case of alpha 1-antitrypsin deficiency associated with hypogammaglobulinemia and recurrent pulmonary thrombosis

    Raghav Gupta


    Full Text Available Alpha 1-antitrypsin (AAT belongs to the family of serpins (serine protease inhibitors. Loop sheet polymerization is the pathology behind serpinopathies which encompasses AAT, anti-thrombin III and neuroserpin deficiency. To the best of our knowledge, we report the first case of alpha 1-antitrypsin deficiency associated with hypogammaglobulinemia and recurrent pulmonary thrombosis without any concomitant use of drugs.

  1. Deficiency of the natural anticoagulant proteins in women with pregnancy related venous thromboembolism

    Mitić Gorana


    Full Text Available Inherited thrombophilia can be defined as a predisposition to thrombosis caused by heritable defects, such as mutations in genes encoding the natural anticoagulants or clotting factors. Pregnancy related risk of VTE is sixfold increased comparing to non pregnant age matched women. Pregnancy is an independent risk factor for the development of venous thromboembolism and this risk is further increased by the presence of thrombophilia. Aim of the study: The aim of the study was to evaluate the association between deficiency of natural anticoagulants: antithrombin, protein C and protein S and pregnancy related thromboembolism. We have determined the activities of antithrombin, proten C and protein S in 74 women with pregnancy related thrombosis and in 45 healthy women who had at least two uncomplicated pregnancies. Among the women with the history of venous thromboembolism antithrombin deficiency was found in 4 (5.4%, protein C deficiency in 2 (2.7% and protein S deficiency in 5 (6.76%. The total of 11 (14.6% women was found to be deficient. Not a single woman in the control group was found to be deficient in natural anticoagulants. Deficiencies of coagulation inhibitors are associated with an increased risk of venous thrombosis during pregnancy and puerperium (p= 0.006. Antithrombin, protein C and protein S deficient women are at higher risk of developing venous thromboembolism during antepartal period (p= 0.0097. Prophylactic treatment with heparin should be recommended from the very beginning of the following pregnancy in women with antithrombin, protein C or protein S deficiency.

  2. [Congenital thrombophilia].

    Kojima, Tetsuhito


    Congenital thrombophilia is a thrombotic diathesis caused by a variety of genetic abnormalities in blood coagulation factors or their inhibitory factors associated with physiological thrombus formation. Patients with congenital thrombophilia often present with unusual clinical episodes of venous thrombosis (occasionally combined with pulmonary embolism, known as venous thromboembolism) at a young age and recurrence in atypical vessels, such as the mesenteric vein and superior sagittal sinus, often with a family history of this condition. Studies in Japan as well as in western countries have shown congenital thrombophilia to be caused by a wide variety of genetic abnormalities in natural anticoagulant proteins, such as antithrombin, protein C, and protein S. However, there may still be many unknown causes of hereditary thrombosis. We recently reported a case of hereditary thrombosis induced by a novel mechanism of antithrombin resistance, that is, congenital thrombophilia caused by a gain-of-function mutation in the gene encoding the coagulation factor prothrombin. PMID:27076244

  3. Effect of a single plasma transfusion on thromboembolism in 13 dogs with primary immune-mediated hemolytic anemia.

    Thompson, Mary F; Scott-Moncrieff, J Catharine; Brooks, Marjory B


    Thirteen dogs with primary immune-mediated hemolytic anemia received fresh-frozen plasma within 12 hours of admission, in addition to unfractionated heparin and other therapies, such as prednisone, azathioprine, and packed red blood cell transfusion. Antithrombin activity was quantified prior to transfusion and at 30 minutes and 48 hours after transfusion. Plasma antithrombin activity did not change significantly after a single plasma transfusion. There were no deaths in the first 48 hours of treatment. Thromboembolism was identified at necropsy in six of 10 dogs that died within 12 months of admission. There was no significant difference in the incidence of thromboembolism between the current treatment group and a historical control group. PMID:15533964

  4. Localization of blood coagulation factors in the germinal centers of human Peyer's patches

    Kudo, S.; Yamakawa, Mitsunori; Imai, Yutaka; Tsukamoto, M.


    The immunohistochemical distribution of 15 blood coagulation factors in the germinal centers (GCs) of human Peyer's patches (PPs) was studied. Although factor VIII, active alpha-thrombin, and fibrinogen were hardly evident in the GCs, the majority of coagulation factors, such as kallikrein, high-molecular-weight kininogen, factos XII, X, IX, VII, V, XIIIa and XIIIb, prothrombin, anti-thrombin 111 and inactive alpha-thrombin were found, showing a lace-like s...

  5. Multiple presence of prothrombotic risk factors in Croatian children with arterial ischemic stroke and transient ischemic attack

    Leniček Krleža, Jasna; Đuranović, Vlasta; Bronić, Ana; Coen Herak, Desiree; Mejaški-Bošnjak, Vlatka; Zadro, Renata


    Aim To determine the frequency of inherited and acquired prothrombotic risk factors in children with arterial ischemic stroke (AIS) and transient ischemic attacks (TIA) in Croatia. Methods We investigated 14 prothrombotic risk factors using blood samples from 124 children with AIS or TIA and 42 healthy children. Prothrombotic risk factors were classified into five groups: natural coagulation inhibitors (antithrombin, protein C, protein S), blood coagulation factors (FV Leiden and FII 20210), ...

  6. Changes in Blood Parameters and the Expression of Coagulation-Related Genes in Lactating Sprague–Dawley Rats

    Urasoko, Yoshinaka; He, Xi Jun; Masao, Takano; KINOSHITA, YUICHI; Edamoto, Hiroshi; Hatayama, Kazuhisa; Asano, Yuzo; Tamura, Kazutoshi; Mochizuki, Masahiro


    This study measured blood parameters, particularly those related to coagulation, and alterations in the expression levels of blood-coagulation–related genes in lactating Sprague–Dawley rats. The day of delivery was designated as lactation day 0 (LD 0). On the day after delivery (LD 1), prothrombin time and overall activity of vitamin-K–dependent coagulation factors were decreased, whereas fibrinogen contents, platelet counts and antithrombin III concentrations were increased as compared with ...

  7. Evaluation of the plasma quality after filtration

    M Mahmoodian Shooshtari; Mousavi Hosseini, K.


    "n  "n "nBackground and the purpose of the study: The quality of some of the human plasma derived drugs such as coagulation factor VIII and coagulation factor IX which can be used for the treatment of hemophilia A and B, depends on their activity which may be affected by filtration. In this study the quality of plasma with respect to coagulation factors FVII, FVIII, FIX, FV, FXI, Fibrinogen, antithrombin III, anti-plasmin and antitrypsin activities obtained after plasm...

  8. inherited thrombophilia and recurrent pregnancy loss Trombofilias heredadas y pérdida gestacional recurrente

    Angela Patricia Cadavid Jaramillo; Wálter Darío Cardona Maya; Serguei Abel Castañeda Ospina


    One of the causes of recurrent pregnancy loss is thrombophilia, defined as a tendency to thrombosis or hypercoagulability, with various clinical manifestations dependent on the vascular region affected by the absence of blood flow. Thrombophilias can be classified as inherited or acquired, according to the nature of their cause. The former are factor V Leiden, the prothrombin G20210A, the methilenetetrahydrofolate reductase C677T, deficiencies of the natural anticoagulants antithrombin III, p...

  9. Platelet and coagulation factors in proliferative diabetic retinopathy.

    Borsey, D. Q.; Prowse, C. V.; Gray, R S; Dawes, J.; James, K.; Elton, R A; Clarke, B F


    Plasma beta-thromboglobulin, platelet factor 4, fibrinogen, fibrinopeptide A, antithrombin III, factor VIII related antigen, alpha 2-macroglobulin, platelet count, and total glycosylated haemoglobin were measured in three well matched groups of subjects: non-diabetic controls, diabetics without retinopathy, and diabetics with proliferative retinopathy. beta-thromboglobulin and platelet factor 4 concentrations were significantly higher in the diabetics with retinopathy than in the controls and...

  10. Effects of monophasic low-dose oral contraceptives on fibrin formation and resolution in young women

    Petersen, K R; Sidelmann, Johannes Jakobsen; Skouby, S O;


    concentration of plasminogen activator inhibitor. Thrombin-antithrombin III complexes and fibrin degradation products were unchanged, signifying no effect of hormonal intake on the degree of activation of the coagulation system or the efficacy of fibrinolysis. CONCLUSION: The overall dynamic balance between...... generation and resolution of fibrin was maintained during treatment with both hormonal compounds. Our findings suggest that the risk of thrombosis in normal women should not be increased....

  11. Den haemostatiske balance under behandling med nyere p-pillepraeparater

    Petersen, K R; Skouby, S O; Sidelmann, Johannes Jakobsen;


    concentration of tissue plasminogen activator inhibitor. The ratio between thrombin-antithrombin-III-complexes and fibrin degradation products were unchanged signifying no effect of hormonal intake on the balance between thrombin formation and fibrin resolution. In conclusion, the dynamic balance between...... generation and resolution of fibrin was undisturbed during treatment with both hormonal compounds and our findings do not provide evidence for increased risk of thrombosis in normal women....

  12. High incidence of thrombophilia detected in Chinese patients with venous thrombosis

    Liu, HW; Kwong, YL; BOURKE, C; Lam, CK; Lie, AKW; Wei, D; Chan, LC


    Venous thromboembolism is rare in Chinese. To determine the incidence and disease profile of thrombophilia in Chinese patients with thrombosis, 52 unselected Chinese patients with documented venous thrombosis were studied for the presence of thrombophilia. Levels of antithrombin III (AT III), protein C (PC) and protein S (PS) as well as the presence of acquired lupus anticoagulant (LA) and anticardiolipin antibody (ACA) were investigated. Thirty patients were found to be abnormal. These consi...

  13. Aptamer Based Microsphere Biosensor for Thrombin Detection

    Xudong Fan; White, Ian M.; Suter, Jonathan D.; Hongying Zhu


    We have developed an optical microsphere resonator biosensor using aptamer as receptor for the measurement of the important biomolecule thrombin. The sphere surface is modified with anti-thrombin aptamer, which has excellent binding affinity and selectivity for thrombin. Binding of the thrombin at the sphere surface is monitored by the spectral position of the microsphere's whispering gallery mode resonances. A detection limit on the order of 1 NIH Unit/mL is demonstrated. Control experiments...

  14. Functional domains of rabbit thrombomodulin.

    Bourin, M C; Boffa, M C; Björk, I.; Lindahl, U


    Thrombomodulin isolated from rabbit lung was separated by ion-exchange chromatography on DEAE-cellulose into a retarded (acidic) and a nonretarded (nonacidic) fraction. Both fractions contained the cofactor required for the activation of protein C. In addition, the acidic fraction (but not the nonacidic fraction) prevented the clotting of fibrinogen by thrombin ("direct" anticoagulant activity) and accelerated the inhibition of thrombin by antithrombin (effect corresponding to 2-10 internatio...

  15. The Levels of Tissue Factor Pathway Inhibitor in Sepsis Patients Receiving Prophylactic Enoxaparin

    Hadil A. Al Otair


    Full Text Available Objective: Sepsis syndrome is usually accompanied by activation of blood coagulation mechanisms. Earlier studies found deficiencies of the 3 main natural anticoagulants, antithrombin, protein C, and protein S. However, none of these inhibitors block tissue factor, the prime trigger of coagulation during sepsis that is controlled specifically by the tissue factor pathway inhibitor (TFPI. The aim of this study was to characterize the fluctuations in the levels of natural anticoagulants, particularly TFPI, in the course of sepsis and to find out their association with the anticoagulant action of the lowmolecular-weight heparin enoxaparin. Materials and Methods: We studied 51 consecutive patients with sepsis. Blood samples were collected from patients at baseline (0 h and at 4, 12, and 24 h after enoxaparin administration. The following assays were undertaken using commercial kits: activated partial thromboplastin time, prothrombin time, thrombin time, total and free TFPI, protein C and protein S, antithrombin, fibrinogen, and anti-factor Xa. Results: Before enoxaparin administration, there was significant prolongation of the prothrombin time and activated partial thromboplastin time, and this remained the case in the 3 subsequent samples. There was marked reduction in the levels of antithrombin, protein C, and total and free protein S to below control values throughout the study. In contrast, plasma levels of both total and free TFPI were markedly elevated and increased after enoxaparin therapy. Anti-factor Xa levels were within the therapeutic range throughout. There was no difference in TFPI levels between those patients who died and those who survived. Conclusion: Sepsis triggered marked release of TFPI from endothelial cells. This persisted and was increased further following the administration of enoxaparin. In contrast, there was marked consumption of the natural coagulation inhibitors antithrombin, protein C, and protein S. These results go


    T.V. Simonyan; V.F. Kirichuk


    The research purpose is focused on studying the role of protein С system in coagulation component disturbance of hemostasis and fibrinolysis system in patients with different forms of gingivitis. The decrease of protein С and antithrombin inactivity, the decrease ofplasminogen reserve in blood, thrombin time shortening and the resistance of factor Va Via to protein Care watched in patients with different forms of gingivitis. The most marked changes in protein С system may be observed in exace...

  17. Haematological stress syndrome in atherosclerosis.

    Stuart, J; George, A J; Davies, A. J.; Aukland, A; Hurlow, R. A.


    Forty patients with atherosclerotic peripheral vascular disease, as compared to 29 healthy controls, showed a significant increase in platelet number and activity, a neutrophil leucocytosis, and a raised level of several acute-phase reactant proteins (fibrinogen, antithrombin III, factor VIII, and serum globulin). The hyperproteinaemia was associated with increases in plasma-, serum-, and blood-viscosity and is the likely cause of the hyperviscosity of vascular disease. These multiple haemost...

  18. Structural Determinants of the Capacity of Heparin to Inhibit the Formation of the Human Amplification C3 Convertase

    Kazatchkine, Michel D.; Fearon, Douglas T.; Metcalfe, Dean D.; Rosenberg, Robert D.; Austen, K. Frank


    The ability of heparin glycosaminoglycan to prevent formation of the properdin-stabilized amplification C3 convertase is independent of antithrombin binding activity and requires substitution of the amino sugar and a degree of oxygen (O)-sulfation which could be on the uronic acid or the amino sugar. Preparations of heparin glycosaminoglycan isolated by different techniques from different species (rat, human, and porcine) exhibited an equivalent capacity to inhibit generation of the amplifica...

  19. Preeclampsia – will Orphan Drug Status facilitate innovative biological therapies?

    Sinuhe eHahn


    It is generally accepted that development of novel therapies to treat pregnancy-relates disorders, such as preeclampsia, is hampered to the paucity of research funding. Hence, it is with great interest to become aware of at least three novel therapeutic approaches for the treatment of this disorder, exploiting either the anticoagulant activity of antithrombin, the free radical scavenging activity of alpha-1-microglobulin, or the regenerative capacity of placenta-derived mesenchymal stem cells...

  20. Preeclampsia – Will Orphan Drug Status Facilitate Innovative Biological Therapies?

    Hahn, Sinuhe


    It is generally accepted that the development of novel therapies to treat pregnancy-related disorders, such as preeclampsia, is hampered by the paucity of research funding. Hence, it is with great interest to become aware of at least three novel therapeutic approaches for the treatment of this disorder: exploiting either the anticoagulant activity of antithrombin, the free radical scavenging activity of alpha-1-microglobulin, or the regenerative capacity of placenta-derived mesenchymal stem c...

  1. Hydrogels by irradiation of a synthetic heparinoid polyelectrolyte

    Sederel, L.C.; Does, van der, Leen; Euverman, B.J.; Bantjes, A.; Kluft, C.; Kempen, H.J.M.


    Gamma irradiation of aqueous solutions of a synthetic heparinoid polyelectrolyte results in the formation of hydrogels, varying in water content and mechanical strength. The equilibrium water content and the mechanical strength of the hydrogels are dependent on the initial polyelectrolyte concentration, the molecular weight of the polyelectrolyte, the percentage of double bonds in the polyelectrolyte and the radiation dose. The polyelectrolyte hydrogels do not deplete Antithrombin III from bl...

  2. Contraception and Thrombophilia - A statement from the German Society of Gynecological Endocrinology and Reproductive Medicine (DGGEF e. V.) and the Professional Association of the German Gynaecologists (BVF e. V.)

    Rabe T; Luxembourg B; Ludwig M.; Dinger JC; Bauersachs R; Rott H; Mueck AO; Albring C


    Venous thromboembolism (VTE) is responsible for more than half a million deaths annually in the European Union, most in older people following surgery, but some in women of reproductive age using various hormonal contraceptives. In some parts of the population inherited defects of the blood coagulation system (factor V Leiden, prothrombin G20210A, protein C, protein S and antithrombin deficiency) are responsible for an increased risk of VTE, which is also influenced by concomitant factors:...

  3. In vivo antithrombotic properties of a heparin from the oocyte test cells of the sea squirt Styela plicata(Chordata-Tunicata)

    L. Cardilo-Reis; M.C.M. Cavalcante; C.B.M. Silveira; M.S.G. Pavão


    In the ascidian Styela plicata, the oocytes are surrounded by two types of accessory cells named follicle cells and test cells. A heparin-like substance with an anticoagulant activity equivalent to 10% of mammalian heparin and about 5% as potent as the mammalian counterpart for the inhibition of thrombin by antithrombin was isolated from the oocyte test cells. In the present study, we compared the antithrombotic and hemorrhagic effects of sea squirt oocyte test cell heparin with those of porc...

  4. [Hypercoagulable workup in ophthalmology. When and what].

    Muñoz-Negrete, F J; Casas-Lleras, P; Pérez-López, M; Rebolleda, G


    Most ophthalmologic disorders secondary to hypercoagulabe state are due to the confluence of congenital and adquired factors. A systematic workup is mandatory. Most of congenital coagulation disorders cause venous trombosis and are inherited autosomal dominantly. In order of frequency these are factor V Leiden mutation (activated protein C resistance), G20210A mutation of the prothrombin gen and protein C, protein S, and antithrombin III deficiencies. Sickle cell anemia can determine arerial and venous thrombosis. In relation with arterial occlusion, the markers most frequently involved are homcysteine fasting levels and the markers of antiphospholipid antibody syndrome. Both of them can also determine venous thrombosis. Several acquired factors can lead to hypoercoagulable state, especially hyperhomocysteinemia, antiphospholipid antibody syndrome, hepatic disease, alcohol and tobacco intake, oral contraceptives, immobilization, surgeries and malignancies. In central venous occlusion is only necessary to rule out hyperhomocysteinemia and antiphospholipid antibody syndrome in young patients without known risk factors. In central artery occlusion, hypercoagulable workup is only recommended for patients less than 50 years-old with unknown emboli source. In this cases protein C, protein S, and antithrombin III deficiencies, homocystein, sickle cell disease and antiphospholipid antibody syndrome will ruled out. In non arteritic ischemic optic neuropathy hypercoagulable work up is not necessary. In amaurosis fugax without known emboli source, it is recommended to rule out etiologies of arterial occlusion, especially antithrombin III deficiencies, homocystein, sickle cell disease and antiphospholipid antibody syndrome. PMID:19658050

  5. sEPCR Levels in Chronic Myeloproliferative Diseases and Their Association with Thromboembolic Events: A Case-Control Study

    Figen Atalay


    Full Text Available OBJECTIVE: Venous, arterial, and microcirculatory events are frequently encountered in the clinical course of essential thrombocytosis and polycythemia vera. We aimed to investigate the levels of soluble endothelial protein C receptor (sEPCR in myeloproliferative diseases to see whether there was a difference between the patients with and without history of thromboembolism. METHODS: The study included patients with polycythemia vera (n=12, patients with essential thrombocytosis (n=13, and controls (n=29. In all groups, we measured proteins C and S, antithrombin and sEPCR levels, and plasma concentrations of thrombin-antithrombin complex, prothrombin fragments 1+2, and D-dimer. RESULTS: Comparing the patients with and without history of thromboembolic attack, statistically significant differences were not observed in terms of sEPCR, D-dimer, thrombin-antithrombin complex, prothrombin fragments 1+2, and hematocrit levels (p=0.318, 0.722, 0.743, 0.324, and 0.065, respectively. CONCLUSION: Significant increase in the parameters that reflect activation of coagulation, such as sEPCR, thrombinantithrombin complex, prothrombin fragments 1+2, and D-dimer, reflects the presence of a basal condition that leads to a tendency toward thrombosis development in ET and PV when compared to healthy controls.

  6. Selected thrombosis and atherosclerosis risk factors in children with idiopathic nephrotic syndrome

    Beata Bieniaś


    Full Text Available The purpose of our study was to evaluate selected thrombosis and atherosclerosis risk factors in children with idiopathic nephrotic syndrome (INS at three  stages of the disease (I – in acute phase before steroid therapy, II – during steroid therapy after resolution of proteinuria, III – in remission after completion of steroid therapy.In all children, serum total homocysteine, lipoprotein (a, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol and triglycerides levels were measured at three stages of the disease. Plasma antithrombin III, fibrinogen and D-dimer levels were also determined.  At all stages of INS, the serum t-HCY levels were similar and significantly higher than in controls.  Serum lipoprotein (a level, plasma antithrombin III, fibrinogen and D-dimer levels were significantly higher at stage I than at stages II, III and controls.In conclusion, children with INS are at high risk of thrombosis and atherosclerosis. Keywords: Idiopathic nephrotic syndrome, Homocysteine, Lipoprotein (a, Antithrombin III, fibrinogen, D-dimer

  7. Evaluation of the plasma quality after filtration

    M Mahmoodian Shooshtari


    Full Text Available "n  "n "nBackground and the purpose of the study: The quality of some of the human plasma derived drugs such as coagulation factor VIII and coagulation factor IX which can be used for the treatment of hemophilia A and B, depends on their activity which may be affected by filtration. In this study the quality of plasma with respect to coagulation factors FVII, FVIII, FIX, FV, FXI, Fibrinogen, antithrombin III, anti-plasmin and antitrypsin activities obtained after plasma filtration with CPD (citrate-phosphate-dextrose using integral filter was evaluated. "nMethods:Sixty units of plasma were individually separated from whole blood by centrifugation and immediately filtered by integral filter system. Specific plasma filtration was carried out between 4 and 20 hrs after blood donation. Before filtration, 60 units of non filtered fresh plasmas were kept as control. "nCoagulation factors were determined by one-stage clotting assay in an automated system. Antithrombin III activity was determined by immunochrom assay in an automated system. Activity of anti-plasmin was determined by Berichrom α2 - antiplasmin and antitrypsin activity was assayed with human neutrophil elastase. "n  "nResults:The activity of coagulation factors FVIII, FIX, Fibrinogen, FV, and FXI, were not affected by filtration, in all experiments. Filtration only caused negligible change in FVII activity. Antithrombin III, anti-plasmin and antitrypsin activities were not influenced by filtration. Non-filtrated and filtrated plasma values were not significantly different (P> 0.05. Conclusions:Plasma filtration dose not result in a measurable impairment of coagulation factors and inhibitors. Although a little changes in FVII activity was observed after filtration, but these filtration-dependent changes apparently have no impact on the therapeutic quality of whole blood- filtered fresh plasma for transfusion.

  8. Test of hirudin activity by tracking the binding of hirudin to thrombin in the presence of BS3 cross-linking.

    Liu, Yanfang; Yang, Jian; Wang, Jiangmin; Huang, Qingmei; Yang, Xiaohong; Zhang, Jianhua


    Hirudin has a great potential in inhibiting thrombin, and its antithrombin activity has direct bearing on its clinical application. Using bovine alpha-thrombin and recombinant hirudin of Poecilobdella javanica purified from Phichia pastoris as materials, this study introduced a novel method to testing antithrombin activity of hirudin visually and dynamically by tracking the binding of hirudin to thrombin. After incubating the mixture of thrombin and hirudin at 37 °C for 5 min, the binding of hirudin to thrombin was cross-linked by bis[sulfosuccinimidyl] suberate for 30 min and visualized by SDS-polyacrylamide gel electrophoresis. With the aid of image analysis on the basis of INRA-Noésis E1D analysis software, antithrombin activity of hirudin was calculated through intensity variations of protein bands of either thrombin-hirudin compound, unbound thrombin, or unbound hirudin. In this regard, activity of the given hirudin was tested to be 5625 ATU/mg based on a single reaction, and 5675.3 ATU/mg based on a series of reactions in a stepwise manner, close to the result of 6000 ATU/mg concluded by titration method. The superiorities of the method include good accuracy (the minimum testable concentration of hirudin is 1.5 μg/ml) and little sample consumption (sample consumption of hirudin is generally 1-11.5 μl using the apparatus of Mini Protean 3 Cell). Easy operation, low input, and equipment requirement also grant it as an effective way. PMID:26332983

  9. Human recombinant interleukin-1 beta- and tumor necrosis factor alpha-mediated suppression of heparin-like compounds on cultured porcine aortic endothelial cells

    Cytokines are known to tip the balance of the coagulant-anticoagulant molecules on the endothelial cell surface toward intravascular coagulation. Their effects on endothelial cell surface-associated heparin-like compounds have not been examined yet. Incorporation of [35S]sulfate into heparan sulfate on cultured porcine aortic endothelial cells was suppressed by human recombinant interleukin-1 beta (rIL-1 beta) or tumor necrosis factor alpha (rTNF alpha) in a dose- and time-dependent manner with little effect on cell number, protein content, and [3H]leucine incorporation of cells. Maximal inhibition was achieved by incubation of cells with 100 ng/ml of rIL-1 beta or 5 ng/ml of rTNF alpha for 12-24 hours, resulting in a reduction of the synthesis of heparan sulfate on the cell surface by approximately 50%. The dose dependency was consistent with that seen in the stimulation of endothelial cell procoagulant activity by each cytokine. The suppression of heparan sulfate synthesis was sustained for at least 48 hours after pretreatment of cells with cytokines and was unchanged after the addition of indomethacin or polymyxin B. The rate of degradation of prelabeled 35S-heparan sulfate on the cell surface was not altered by cytokine treatments. Neither the size, the net negative charge, nor the proportion of the molecule with high affinity for antithrombin III of endothelial cell heparan sulfate was changed by cytokines. Furthermore, specific binding of 125I-labeled antithrombin III to the endothelial cell surface was reduced to 40-60% of control by cytokines. In parallel with reduction in binding, antithrombin III cofactor activity was partially diminished in cytokine-treated endothelial cells. Thus, cytokine-mediated suppression of heparin-like substance on endothelial cells appears to be another cytokine-inducible endothelial effects affecting coagulation

  10. Thrombelastography and biomarker profiles in acute coagulopathy of trauma: a prospective study

    Larsen Claus F


    Full Text Available Abstract Background Severe injury induces an acute coagulopathy associated with increased mortality. This study compared the Thrombelastography (TEG and biomarker profiles upon admission in trauma patients. Methods Prospective observational study of 80 trauma patients admitted to a Level I Trauma Centre. Data on demography, biochemistry including standard coagulation tests, hematology, transfusions, Injury Severity Score (ISS and TEG were recorded. Retrospective analysis of thawed plasma/serum for biomarkers reflecting tissue injury (histone-complexed DNA fragments, sympathoadrenal activation (adrenaline, noradrenaline, coagulation activation/inhibition and fibrinolysis (sCD40L, protein C, activated Protein C, tissue-type plasminogen activator, plasminogen activator inhibitor-1, D-dimer, prothrombinfragment 1+2, plasmin/α2-antiplasmin complex, thrombin/antithrombin complex, tissue factor pathway inhibitor, antithrombin, von willebrand factor, factor XIII. Comparison of patients stratified according to ISS/TEG maximum clot strength. Linear regression analysis of variables associated with clot strength. Results Trauma patients had normal (86%, hypercoagulable (11% or hypocoagulable (1% TEG clot strength; one had primary hyperfibrinolysis. Hypercoagulable patients had higher age, fibrinogen and platelet count (all p 10 red blood cells the initial 24 h. Patients with normal or hypercoagulable TEG clot strength had comparable biomarker profiles, but the few patients with hypocoagulable TEG clot strength and/or hyperfibrinolysis had very different biomarker profiles. Increasing ISS was associated with higher levels of catecholamines, histone-complexed DNA fragments, sCD40L, activated protein C and D-dimer and reduced levels of non-activated protein C, antithrombin, fibrinogen and factor XIII (all p 26. In patients with ISS > 26, adrenaline and sCD40L were independently negatively associated with clot strength. Conclusions Trauma patients displayed

  11. Influence of dabigatran and rivaroxaban on routine coagulation assays. A nationwide Belgian survey.

    Van Blerk, Marjan; Bailleul, Els; Chatelain, Bernard; Demulder, Anne; Devreese, Katrien; Douxfils, Jonathan; Jochmans, Kristin; Mullier, François; Wijns, Walter; Soumali, Mohamed Rida; Coucke, Wim; Vernelen, Kris; Van de Walle, Philippe


    The Belgian national External Quality Assessment Scheme performed a nationwide survey using lyophilised plasma samples spiked with dabigatran or rivaroxaban to demonstrate to the Belgian clinical laboratories how these drugs affect their routine coagulation assays prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen and antithrombin. Virtually all Belgian laboratories performing routine coagulation testing (189/192) participated in the survey. Both, dabigatran and rivaroxaban significantly prolonged the PT and aPTT in a concentration- and reagent-dependent manner. PT reagents were more influenced by rivaroxaban than by dabigatran and aPTT reagents more influenced by dabigatran than by rivaroxaban. Among PT reagents, Neoplastin R® was the most sensitive to rivaroxaban and Innovin® and Thromborel S® the least sensitive. Converting PT results to INR only increased the variability between reagents. Among aPTT reagents, Actin FSL® was the least sensitive to dabigatran while the other aPTT reagents showed slightly higher sensitivities. The presence of dabigatran led to falsely reduced fibrinogen concentrations when measured with a low thrombin concentration reagent. The presence of dabigatran caused an overestimation of the antithrombin level when measured with a thrombin-based activity assay and the presence of rivaroxaban an overestimation of the antithrombin level when measured with a FXa-based assay. Instrument-related differences were found for all tested parameters. In conclusion, this paper provides detailed information on the effect of dabigatran and rivaroxaban on routine coagulation assays as performed with a large number of reagent/instrument combinations. PMID:25231101

  12. Cell-based laboratory evaluation of coagulation activation by antineoplastic drugs for the treatment of lymphoid tumors

    Tsunaka, Misae; Arai, Reina; Ohashi, Ayaka; Koyama, Takatoshi


    Objectives: Combining vorinostat, L-asparaginase, and doxorubicin (Dox) led to improved response rates in the treatment of lymphoid tumors. However, deep-vein thrombosis has been noted as one of the most serious side effects with these drugs, and how these regimens cause deep-vein thrombosis is unclear. Methods: We investigated the procoagulant effects of vorinostat, L-asparaginase, and doxorubicin in lymphoid tumors, focusing on tissue factor, phosphatidylserine, and antithrombin. The human vascular endothelial cell line EAhy926 as well as the lymphoid neoplastic cell lines HUT78 (cutaneous T-cell lymphoma), Molt4 (acute T-lymphoblastic leukemia), and Ramos (Burkitt lymphoma) were employed to investigate these procoagulant effects. Results: Vorinostat, L-asparaginase, and doxorubicin induced exposure of phosphatidylserine and procoagulant activity on the surface of lymphoid tumor cells. Vorinostat and doxorubicin also induced phosphatidylserine exposure and increased procoagulant activity on EAhy926 cells. Expression of tissue factor antigen was induced by doxorubicin on the surface of each type of cells, whereas expression of tissue factor mRNA was unchanged. Secretion of antithrombin from HepG2 cells was reduced only by L-asparaginase. Conclusion: These data suggest that vorinostat and doxorubicin may induce procoagulant activity in vessels through apoptosis of tumor cells and through phosphatidylserine exposure and/or tissue factor expression on vascular endothelial cells. L-asparaginase may induce a thrombophilic state by reducing the secretion of anticoagulant proteins such as antithrombin. The laboratory methods described here could be useful to evaluate the procoagulant effects of antineoplastic drugs. PMID:27504186

  13. Effect of whole-body irradiation on the level of blood clotting factors in rats

    Rats were irradiated with 6.0, 7.0 or 9.5 Gy of X-rays. Blood was collected on the 1,3,7,14 or 30th day after exposure. Fibrinogen, thrombin, prothrombin, factor X and antithrombin 3 levels were determined in blood plasma. In irradiated rats the level of these factors underwent a change which depended to some extent on the radiation dose absorbed. In general, there was a temporary elevation of all factors, lasting longer when the dose applied was higher. 24 refs., 6 tabs. (author)

  14. Synthetic oligosaccharides as heparin-mimetics displaying anticoagulant properties.

    Avci, Fikri Y; Karst, Nathalie A; Linhardt, Robert J


    Heparin and low molecular weight heparins are major clinical anticoagulants and the drugs of choice for the treatment of deep venous thrombosis. The discovery of an antithrombin binding domain in heparin focused interest on understanding the mechanism of heparin's antithrombotic/ anticoagulant activity. Various heparin-mimetic oligosaccharides have been prepared in an effort to replace polydisperse heparin and low molecular weight heparins with a structurally-defined anticoagulant. The goal of attaining a heparin-mimetic with no unwanted side-effects has also provided motivation for these efforts. This article reviews structure-activity relationship (SAR) of structurally-defined heparin-mimetic oligosaccharides. PMID:14529394

  15. Range of fractionated plasma products to optimize plasma resources

    Thierry Burnouf


    @@ HUMAN PLASMA is a source material that is crucial for the production of unique therapeutic fractionated products. Indeed, plasma contains hundreds of proteins ensuring many physiological functions. The most abun-dant proteins, albumin and immunoglobulin G (IgG) ,are present at about 35 and 10 g/L,respectively,repre-senting about 80% of all plasma proteins. However,other important therapeutic proteins include the coagu-lation factors (factor Ⅷ (F Ⅷ) ; FIX ; Von Willebrand Factor (VWF), fibrinogen) various protease inhibitors (alpha 1-antitrypsin ; antithrombin; C1-esterase) and anticoagulants (protein C) which exhibit potent physi-ological activity.

  16. Assessing the relative importance of the biophysical properties of amino acid substitutions associated with human genetic disease

    Terp, Bent N; Cooper, David N; Christensen, Inge T; Jørgensen, Flemming Steen; Bross, Peter Gerd; Gregersen, Niels; Krawczak, Michael


    five human genes encoding arylsulphatase A (ARSA), antithrombin III (SERPINC1), protein C (PROC), phenylalanine hydroxylase (PAH), and transthyretin (TTR). These proteins were chosen on the basis of 1) the availability of a crystallographic structure, and 2) a sufficiently large number of amino acid...... replacements being logged in HGMD. A total of 9,795 possible mutant structures were modeled and 20 different biophysical parameters assessed. Together with the HGMD-derived spectra of clinically detected mutations, these data allowed maximum likelihood estimation of RCOL profiles for the 20 parameters studied...


    Researchers at the Eunice Kennedy Shriver National Institute of Child Health and Human Development (“NICHD”), seek CRADA partner or collaboration for development of agents to treat multiple sclerosis or other conditions associated with myelin remodeling by administering an agent that inhibits cleavage of Neurofascin 155 or Caspr1. The agent could be a thrombin inhibitor, an agent that inhibits thrombin expression, an anti-thrombin antibody that specifically inhibits thrombin mediated cleavage of Neurofascin 155, a mutated version or fragment of Neurofascin 155 or Caspr1, or antibodies to Neurofascin 155 or Caspr1.

  18. Purpura fulminans in a patient with mixed connective tissue disease.

    Murad, Aizuri A


    A 43-year-old lady was admitted to the intensive care unit with sepsis. She had a history of mixed connective tissue disease, Raynaud\\'s syndrome and hypothyroidism. 2 days later, she developed a purpuric rash on her face and extremities with a livedoid background. Few days later, her distal fingers and toes became gangrenous which then had to be amputated. Laboratory investigations showed that she was coagulopathic and had multiple organ dysfunctions. Antiphospholipid antibodies were negative; however, protein C and antithrombin III levels were low. A skin biopsy showed fibrinoid necrosis in the vessel wall with microthrombi and red-cell extravasation. A diagnosis of purpura fulminans was made.

  19. Kontrazeption & Thrombophilie - Eine Stellungnahme der Deutschen Gesellschaft für Gynäkologische Endokrinologie und Fortpflanzungsmedizin (DGGEF) e. V. und des Berufsverbands für Frauenärzte (BVF) e.V.

    Rabe T; Luxembourg B; Ludwig M; Dinger JC; Bauersachs R; Rott H; Mueck AO; Albring C


    Mehr als eine halbe Million Menschen sterben jährlich in der EU an venösen Thromboembolien (VTE), meist in höherem Alter oder im Zusammenhang mit Operationen, aber auch junge Frauen im reproduktiven Alter unter Anwendung hormonaler Kontrazeptiva. In einigen Teilen der Bevölkerung sind angeborene Störungen im Gerinnungssystem (Faktor-V-Leiden, Prothrombin-Mutation G20210A, Protein C-, Protein S- und Antithrombin-Mangel) für ein erhöhtes VTE-Risiko verantwortlich. Das VTE-Risiko wird durch f...

  20. Cutaneous necrosis in pregnancy secondary to activated protein C resistance in hereditary angioedema.

    Perkins, W; Downie, I; Keefe, M; Chisholm, M


    A 26-year-old woman with hereditary angineurotic oedema (HAE) presented at 22 weeks gestation with severe cutaneous necrosis similar to that seen in coumarin skin necrosis. Protein S deficiency secondary to HAE and pregnancy was postulated. Treatment with heparin, C1-inhibitor concentrates, systemic steroids and surgical debridement resulted in a successful outcome for both mother and child. Subsequent investigations revealed normal levels of protein C, antithrombin III, total protein S, free protein S but reduced function protein S activity with evidence of activated protein C resistance. Cutaneous necrosis has not been reported in associated with activated protein C resistance previously and the possible mechanisms are discussed. PMID:7745572

  1. Thrombotic actions of nonionic intravascular contrast agents

    Experimental studies show that nonionic contrast media (NICM) produce markedly lower or no anticoagulant effects and, on extended contact, can produce procoagulant actions in native blood. Patients with hemostatic imbalance (such as a decrease of antithrombin III, protein C, thrombocytosis, hyperfibrinogenemia, rheologic disorders, hemoconcentration, and dehydration) are at high thrombotic risk during angiography. Exogenous compensation of the hemostatic deficit minimizes the thrombogenic actions of NICMs. Anticoagulants such as hirudin and low-molecular-weight heparin can antagonize the procoagulant actions of NICM. In a primate model (Macaca mulatta), thrombogenic properties of nonionic contrast media have been successfully antagonized with anticoagulant drugs

  2. Hormone replacement therapy and hypercoagulability. Results from the Prospective Collaborative Danish Climacteric Study

    Sidelmann, Johannes Jakobsen; Jespersen, Jørgen; Andersen, Lars F.;


    norethindrone acetate, (E). E(2) combined with local delivery of levonorgestrel and (F). E(2)V plus medroxyprogesterone. HRT-induced changes in the concentration of inhibitors of coagulation and markers of fibrin turnover during 12 months of treatment. RESULTS: Significant decreases of antithrombin and protein......-normalised ratio were measured at baseline and after 6 and 12 months of HRT in six groups of healthy postmenopausal women: (A). no HRT (reference group), (B). continuous oestradiol valerate (E(2)V) plus cyproterone acetate, (C). cyclic E(2)V plus cyproterone acetate, (D). continuous combined oestrogen (E(2)) plus...

  3. Anesthetic management of a child with nephrotic syndrome undergoing open heart surgery: Report of a rare case

    Vishnu Datt


    Full Text Available The congenital nephrotic syndrome (NS in infancy and childhood is an important entity but combination with acyanotic congenital heart disease is uncommon. Anesthesia in such cases is challenging because of associated problems like hypo-protienemia, anti-thrombin III deficiency, edema, hyperlipidemia, coagulopathy, cardiomyopathy, immunodeficiency, increased lung water etc. We describe anesthetic management of a patient with childhood NS and sinus venosus atrial septal defect (ASD undergoing open heart surgery. We also suggest guidelines for safe conduct of anesthesia and CPB in such patients.

  4. Drug: D03674 [KEGG MEDICUS

    Full Text Available D03674 Drug Enoxaparin sodium (JAN/USAN/INN); Lovenox (TN) Antithrombotic intended ...olism 33 Blood and body fluid agents 333 Anticoagulants 3334 Heparins D03674 Enoxaparin sodium (JAN/USAN/INN...ITHROMBOTIC AGENTS B01A ANTITHROMBOTIC AGENTS B01AB Heparin group B01AB05 Enoxaparin D0367...lume Expanders Anticoagulants Enoxaparin D03674 Enoxaparin sodium (JAN/USAN/INN) Target-based classification... of drugs [BR:br08310] Others Heparin binding proteins antithrombin III [HSA:462] [KO:K03911] Enoxaparin [ATC:B01AB05] D0367

  5. Cerebral Vein Thrombosis:Screening of Acquired and Hereditary Thrombophilic Risk Factors

    Sarraf Payam


    Full Text Available Cerebral vein thrombosis (CVT is an infrequent condition with a large variety of causes that can lead to serious disabilities. However, in 20% to 35% of cases, no cause is found. In this study we evaluated the hereditary (P & C Proteins, antithrombin, mutation of prothrombin G20210A and factor V Leiden, other risk factors (hyperhomocycteinemia, factor VIII, ACL-ab, APL-ab, and OCP and clinical manifestations among a population of Iranian patients with CVT. 18 women and 10 men aged 16 to 50 years with CVT were screened for inherited and acquired coagulation risk factors. No one had an abnormal ACL-ab, APL-ab or antithrombin III deficiency. One had prothrombin G20210A mutation (heterozygot (3.6%. Hyperhomocycteinemia was observed in 5 patients (17.9%. APC-R was decreased in 3 (10.7%. 2 had positive factor V Leiden mutation (heterozygot (7.1%. 17 had an increased of factor VIII (60.7. PS and PC deficiencies were each detected in two cases (7.1%. Conclusion: Our study suggests that screening for inherited thrombophilia may be an integral part in the diagnostic workup and duration of treatment in patients with CVT.

  6. New anticoagulants for the prevention of venous thromboembolism

    Cecilia Becattini


    Full Text Available Cecilia Becattini, Alessandra Lignani, Giancarlo AgnelliInternal and Cardiovascular Medicine and Stroke Unit, University of Perugia, ItalyAbstract: Anticoagulant drugs have an essential role in the prevention and treatment of thromboembolic diseases. Currently available anticoagulants substantially reduce the incidence of thromboembolic events in a number of clinical conditions. However, these agents have limitations that strengthen the case for the development of new anticoagulants. An ideal anticoagulant should be at least as effective as those currently in use, as well as safe, simple to use, and widely applicable. The majority of new anticoagulants currently under investigation are small molecules with a selective and direct anti-Xa or antithrombin action, allowing oral administration in fixed doses. These new agents are in different phases of clinical development. The anti-Xa agent rivaroxaban and the antithrombin agent dabigatran are already available for the prophylaxis of venous thromboembolism in some countries. Apixaban is in an advanced phase of clinical development and several anti-Xa agents are currently approaching phase III clinical trials. Promising results in terms of efficacy and safety profiles have been obtained with these agents in different clinical conditions. Differences in pharmacokinetics and pharmacodynamics could offer the potential for individualized anticoagulant therapies in the near future.Keywords: anticoagulant therapy, antithrombotic therapy, anticoagulants, direct thrombin inhibitors, factor Xa inhibitors

  7. New anticoagulants for the prevention of venous thromboembolism

    Becattini, Cecilia; Lignani, Alessandra; Agnelli, Giancarlo


    Anticoagulant drugs have an essential role in the prevention and treatment of thromboembolic diseases. Currently available anticoagulants substantially reduce the incidence of thromboembolic events in a number of clinical conditions. However, these agents have limitations that strengthen the case for the development of new anticoagulants. An ideal anticoagulant should be at least as effective as those currently in use, as well as safe, simple to use, and widely applicable. The majority of new anticoagulants currently under investigation are small molecules with a selective and direct anti-Xa or antithrombin action, allowing oral administration in fixed doses. These new agents are in different phases of clinical development. The anti-Xa agent rivaroxaban and the antithrombin agent dabigatran are already available for the prophylaxis of venous thromboembolism in some countries. Apixaban is in an advanced phase of clinical development and several anti-Xa agents are currently approaching phase III clinical trials. Promising results in terms of efficacy and safety profiles have been obtained with these agents in different clinical conditions. Differences in pharmacokinetics and pharmacodynamics could offer the potential for individualized anticoagulant therapies in the near future. PMID:20531960

  8. Hemostatic system changes predictive value in patients with ischemic brain disorders

    Raičević Ranko


    Full Text Available The aim of this research was to determine the importance of tracking the dynamics of changes of the hemostatic system factors (aggregation of thrombocytes, D-dimer, PAI-1, antithrombin III, protein C and protein S, factor VII and factor VIII, fibrin degradation products, euglobulin test and the activated partial thromboplastin time – aPTPV in relation to the level of the severity of ischemic brain disorders (IBD and the level of neurological and functional deficiency in the beginning of IBD manifestation from 7 to 10 days, 19 to 21 day, and after 3 to 6 months. The research results confirmed significant predictive value of changes of hemostatic system with the predomination of procoagulant factors, together with the insufficiency of fibrinolysis. Concerning the IBD severity and it's outcome, the significant predictive value was shown in the higher levels of PAI-1 and the lower level of antithrombin III, and borderline significant value was shown in the accelerated aggregation of thrombocytes and the increased concentration of D-dimer. It could be concluded that the tracking of the dynamics of changes in parameters of hemostatic system proved to be an easily accessible method with the significant predictive value regarding the development of more severe. IBD cases and the outcome of the disease itself.

  9. Effect of Age on the Hemostatic Function in Patients with Degenerative Diseases of the Large Joints

    Igor L. Shlykov, PhD¹, ScD¹


    Full Text Available Background: Aging is associated with an increased hypercoagulable state. Degenerative diseases of the large joints are also accompanied by increased coagulation activity. We investigated the effect of age on the hemostatic function in patients with osteoarthritis. Material and Methods: The study included 192 patients with osteoarthritis admitted to the clinic for primary hip or knee arthroplasty. The patients were categorized into 5 age groups: the age group under 40 years, the 41–to-50 -year age group, the 51–to-60-year age group, the 61-to-70- year age group, and the age group over 70 years. The general blood clotting tests, platelet number, fibrinogen, antithrombin, protein C, TAT, D-dimer, vonWillebrand factor (vWF, PAI-1, ß-thromboglobulin were determined. Results: Among patients with osteoarthritis, the antithrombin III level significantly decreased by the age of 50; however, above the age of 60 there was a distinct decrease in platelet count, and over the age of 70 the activity of the extrinsic coagulation pathway and the plasminogen level dropped significantly. TAT and D-dimer levels were elevated in most of the patients. Conclusion: The decrease in platelet count coupled with the activity of the extrinsic coagulation pathway in elderly osteoarthritic patients may increase blood loss during total arthroplasty; also, the drop in the anticoagulant and fibrinolytic potential may play a negative role in strengthening the prothrombotic state during the postoperative period.

  10. Bivalirudin in combination with heparin to control mesenchymal cell procoagulant activity.

    Xavier Stephenne

    Full Text Available Islet and hepatocyte transplantation are associated with tissue factor-dependent activation of coagulation which elicits instant blood mediated inflammatory reaction, thereby contributing to a low rate of engraftment. The aim of this study was i to evaluate the procoagulant activity of human adult liver-derived mesenchymal progenitor cells (hALPCs, ii to compare it to other mesenchymal cells of extra-hepatic (bone marrow mesenchymal stem cells and skin fibroblasts or liver origin (liver myofibroblasts, and iii to determine the ways this activity could be modulated. Using a whole blood coagulation test (thromboelastometry, we demonstrated that all analyzed cell types exhibit procoagulant activity. The hALPCs pronounced procoagulant activity was associated with an increased tissue factor and a decreased tissue factor pathway inhibitor expression as compared with hepatocytes. At therapeutic doses, the procoagulant effect of hALPCs was inhibited by neither antithrombin activators nor direct factor Xa inhibitor or direct thrombin inhibitors individually. However, concomitant administration of an antithrombin activator or direct factor Xa inhibitor and direct thrombin inhibitor proved to be a particularly effective combination for controlling the procoagulant effects of hALPCs both in vitro and in vivo. The results suggest that this dual antithrombotic therapy should also improve the efficacy of cell transplantation in humans.

  11. Effects of fused hirudin on activity of thrombin and function of platelets

    SHEN Li; CHEN Shao-ping; CAI Zai-long; YANG Sheng-sheng; QIN Yong-wen


    Objective: To investigate whether fused hirudin peptide has both antithrombin and antiplatelet functions. Methods: The core region of fused hirudin was the C-terminal tail of hirudin(hirudin53-64),which could bind to the anion binding exosite (ABE) of thrombin.Arg-Pro-Pro-Gly-Phe(RPPGF) amino acid sequence,a metabolite of bradykinin,was added to the N-terminus of hirudin53-64.It bound to the active site of thrombin.Additionally,Arg-Gly-Asp(RGD)amino acid sequence,an inibitor of glycoprotein Ⅱb/Ⅲa( GP Ⅱb/Ⅲa) receptor,was linked to C-terminus of hirudin53-64.This 26-animo acid-fused hirudin peptide was artificially synthesized,purified and analysed. Results: Fused hirudin peptide significantly lengthened the activated partial thromboplastin time(APTT),thrombin time(TT)and prothrombin time(PT) and inhibited the amidolytic activity of thrombin.The ADP-induced platelet aggregation was markedly inhibited by fused hirudin peptide. Conclusion: Fused hirudin peptide has activity of antithrombin as well as antiplatelet.Therefore bifunctional anticoagulation peptide has capacity to target various components of haemostatic process and may become more powerful antithrombosis agent.

  12. Interaction of basic amino acids, polypeptides and proteins with heparin

    A study has been made of the relative binding affinities for heparin of L-lysine, L-arginine, poly-L-lysine, poly-L-arginine, protamine, thrombin and antithrombin III. The destruction by different concentrations of organic cations of the heparin-methylene blue complex was determined by visible absorption spectroscopy. The strength of binding of the basic amino acids and polypeptides increased with increasing molecular weight, and decreased at low pH. Pulse radiolysis experiments were used to show the effects of increasing concentrations of L-lysine, poly-L-lysine and protamine sulphate on the reaction of e-sub(aq) with the heparin-methylene blue complex. The results indicate that the binding of basic amino acids and polypeptides to heparin is ionic, and that the enhanced rate of association of antithrombin III to thrombin in the presence of heparin is due to partial or complete charge neutralization of the basic groups in the proteins. (U.K.)

  13. Coagulation inhibitors and activated protein C resistance in recurrent pregnancy losses in Indian women

    P Lalita Jyotsna


    Full Text Available Background: Thrombophilias, both acquired and inherited, have been investigated in the etiopathogenesis of unexplained recurrent pregnancy loss. Aim: To study coagulation inhibitors and activated protein C resistance (APCR in recurrent pregnancy losses (RPL occurring in second and third trimesters. Materials and Methods: A total of 30 pregnant women (group A with two or more recurrent unexplained fetal loses were evaluated for APCR, protein C deficiency, protein S deficiency, antithrombin deficiency, and antiphospholipid antibodies (APLA. Thirty age-matched controls were taken (group B comprising of pregnant women with at least one live issue. Statistical Analysis: Comparisons between two group frequencies and group means were made using Chi square test and Student′s t test, respectively. Results: Protein C and protein S levels were reduced in group A compared with group B and the difference was statistically significant (P=0.005 and P=0.032, respectively. The mean value of antithrombin was slightly reduced in group A compared with group B. APCR was observed in 16.6% cases and 3.3% controls. However, the difference was not statistically significant. APLA was observed in 20% cases and none of the controls. Of these, lupus anticoagulant was positive in 16.6% cases and anticardiolipin antibodies in 10% cases. Combined defects were seen in seven patients. Conclusion: There is a significant risk of RPL in pregnant women with thrombophilias. Therefore, screening for thrombophilias may be justified in pregnant women with unexplained recurrent fetal wastage, especially in second and third trimester.

  14. Arteriovenous thrombosis in chronic renal failure patients receving renal replacement therapy

    To determine the frequency of thrombotic complications and to identify factors associated with arteriovenous thrombosis in patients of chronic renal failure receiving renal replacement therapy. Of the 3000 patients evaluated, 61 End Stage Renal Disease (ESRD) patients on regular dialysis, having recent renal transplant, were selected for the study after informed consent. These patients had arteriovenous thrombosis with temporary central lines thrombosis and vascular access problems. Cases of congenital or acquired thrombotic disorders, e.g. with malignancy, DIC, liver disease, systemic lupus erythematosus or other immunologic diseases, pregnancy or women using oral contraceptives, were excluded. Similarly, patients taking any type of anticoagulant therapy during the preceding one week were not included in the study. Findings were recorded in a structured questionnaire. Laboratory analysis was done after clinical and radiological evaluation. Thrombophilia screening included antithrombin, protein C, protein S deficiencies and lupus anticoagulant. Forty-seven out of 61 patients selected were positive for thrombophilia screening with protein C deficiency in 26.2%, protein S deficiency in 16.3%, antithrombin in 5%, lupus anticoagulant in 13.1% and combined deficiency was observed in 16.3%. Of the 3000 patients, 61 with frequency of 2% were found to be deficient in one or had combined deficiency of these. Thus, the study of ESRD patients presenting with arteriovenous thromboembolism emphasizes the need to reconsider the perception that this clinical entity is rare and requires further studies. (author)

  15. [A case report of hereditary angioedema and studies on the serum components of complement, C1-inactivator and proteinase inhibitors during edema attack].

    Mikami, A; Kohno, M


    Sixteen years old girl was admitted because of for the past ten years' frequent edema attack and abdominal pain. Laboratory examination revealed hypocomplementemia, marked depletion of the fourth component of complement and low level of C1-inactivator. Familial studies revealed that her mother was also hypocomplementemic and in low level of C1-inactivator. Serial studies performed on the alterlation of components of complement, C1-inactivator, alpha 1-antitrypsin, antithrombin III, and alpha 2-macroglobulin during edema attack. The fourth component of complement and C1-inactivator were markedly depleted in remission and attack. Remarkable depletion was found in antithrombin III and esterase inhibition activity of C1-inactivator during attack. In contrast, alpha 1-antitrypsin and alpha 2-macroglobulin did not change. The present study may explain that Hageman factor fragments, activated by C1s, promotes kinin generation via kalikrein activation. And the condition that complete functional deficiency of C1-inactivator was main role in this circuit. Fibrynolysis and late components of complement was less influence on edema attack. PMID:3610041

  16. Hyperprolactinemia during antipsychotics treatment increases the level of coagulation markers

    Ishioka M


    Full Text Available Masamichi Ishioka, Norio Yasui-Furukori, Norio Sugawara, Hanako Furukori, Shuhei Kudo, Kazuhiko Nakamura Department of Neuropsychiatry, Graduate School of Medicine, Hirosaki University, Hirosaki, Japan Objective: The strong association between psychiatric patients who receive antipsychotics and the incidence of venous thromboembolism (VTE is known. Although previous reports suggest that hyperprolactinemia often increases markers of activated coagulation, few studies have examined the direct relationship between the prolactin level elevated by antipsychotics and activated markers of activated coagulation.Method: The participants included 182 patients with schizophrenia (male =89, female =93 who received antipsychotic treatments for at least 3 months. Markers of VTE (D-dimer, fibrin/fibrinogen degradation products, and thrombin–antithrombin complex and serum prolactin concentrations were measured.Results: Prolactin levels were significantly correlated with the logarithmic transformation of the D-dimer (r=0.320, P=0.002 and fibrin/fibrinogen degradation product levels (r=0.236, P=0.026 but not of the thrombin–antithrombin complex level (r=0.117, ns among men. However, no correlations were found between the VTE markers and prolactin levels among women. These results were confirmed using multiple regression analyses that included demographic factors and antipsychotic dosages. Conclusion: The current study indicates that hyperprolactinemia is associated with an increase in markers of activated coagulation among men receiving antipsychotics. This finding clinically implies that monitoring and modulating prolactin levels among men are important to decrease the risk of VTE. Keywords: prolactin, antipsychotics, venous thromboembolism

  17. Biosynthesis of heparin. Effects of n-butyrate on cultured mast cells

    Murine mastocytoma cells were incubated in vitro with inorganic [35S]sulfate, in the absence or presence of 2.5 mM n-butyrate, and labeled heparin was isolated. The polysaccharide produced in the presence of butyrate showed a lower charge density on anion exchange chromatography than did the control material and a 3-fold increased proportion of components with high affinity for antithrombin. Structural analysis of heparin labeled with [3H] glucosamine in the presence of butyrate showed that approximately 35% of the glucosamine units were N-acetylated, as compared to approximately 10% in the control material; the nonacetylated glucosamine residues were N-sulfated. The presence of butyrate thus leads to an inhibition of the N-deacetylation/N-sulfation process in heparin biosynthesis, along with an augmented formation of molecules with high affinity for antithrombin. Preincubation of the mastocytoma cells with butyrate was required for manifestation of either effect; when the preincubation period was reduced from 24 to 10 h the effects of butyrate were no longer observed. A polysaccharide formed on incubating mastocytoma microsomal fraction with UDP-[3H]glucuronic acid, UDP-N-acetylglucosamine, and 3'-phosphoadenylylsulfate in the presence of 5 mM butyrate showed the same N-acetyl/N-sulfate ratio as did the corresponding control polysaccharide, produced in the absence of butyrate. These findings suggest that the effect of butyrate on heparin biosynthesis depends on the integrity of the cell

  18. [The indicators of hemostasis and functions of liver in patients with acute lymphoblastic leucosis under manifestation of disease and induction therapy].

    Tarasova, L N; Vladimirova, S G; Skolskaya, O Yu; Tcherepanova, V V


    The function of liver and synthesized by it components of blood coagulation were studied on sampling of 25 patients with acute lymphoblastic leucosis in the debut of disease and under implementation of induction of remission. Before treatment the hepatotoxicity was detected in 40% of patients. The reliable correlation between level of aspartataminotransferase and number of leucocytes and blasts (rs = 0.401 and rs = 0.406 correspondingly) testifies the role of leucosis cells in functional disorders of liver during this period of disease. Against the background of cytostatic therapy the signs of liver damage were presented in 76% of patients. The maximum expressed hepatotoxicity was observed at 9-14 day of treatment. The decrease of components of hemostasis synthesized in liver was observed at 16-21 day of treatment. During this time the synthesis of factors of thrombin complex, antithrombin III and plasminogen was disordered. The plasminogen also decreased at the expense of its consumption as a result of fibrinolysis activation validated by decreased level of D-dimers. It is proved that in the debut and under treatment acute lymphoblastic leucosis the danger of development of thrombotic hemorrhagic complications is related both with derangement of synthesis of procoagulants, antithrombin III and plasminogen and with their consumption in case of activation of intravascular coagulation with leucosis process and cytostatic. PMID:24006640

  19. Glycosaminoglycans affect the interaction of human plasma kallikrein with plasminogen, factor XII and inhibitors

    Gozzo A.J.


    Full Text Available Human plasma kallikrein, a serine proteinase, plays a key role in intrinsic blood clotting, in the kallikrein-kinin system, and in fibrinolysis. The proteolytic enzymes involved in these processes are usually controlled by specific inhibitors and may be influenced by several factors including glycosaminoglycans, as recently demonstrated by our group. The aim of the present study was to investigate the effect of glycosaminoglycans (30 to 250 µg/ml on kallikrein activity on plasminogen and factor XII and on the inhibition of kallikrein by the plasma proteins C1-inhibitor and antithrombin. Almost all available glycosaminoglycans (heparin, heparan sulfate, bovine and tuna dermatan sulfate, chondroitin 4- and 6-sulfates reduced (1.2 to 3.0 times the catalytic efficiency of kallikrein (in a nanomolar range on the hydrolysis of plasminogen (0.3 to 1.8 µM and increased (1.9 to 7.7 times the enzyme efficiency in factor XII (0.1 to 10 µM activation. On the other hand, heparin, heparan sulfate, and bovine and tuna dermatan sulfate improved (1.2 to 3.4 times kallikrein inhibition by antithrombin (1.4 µM, while chondroitin 4- and 6-sulfates reduced it (1.3 times. Heparin and heparan sulfate increased (1.4 times the enzyme inhibition by the C1-inhibitor (150 nM.

  20. Heparin cofactor II is degraded by heparan sulfate and dextran sulfate.

    Saito, Akio


    Heparan sulfate normally binds to heparin cofactor II and modulates the coagulation pathway by inhibiting thrombin. However, when human heparin cofactor II was incubated with heparan sulfate, heparin cofactor II became degraded. Other glycosaminoglycans were tested, including hyaluronic acid, chondroitin sulfates, dermatan sulfate, and heparin, but only dextran sulfate also degraded heparin cofactor II. Pretreatment of heparan sulfate with heparinase reduced its heparin cofactor II-degrading activity. Heparan sulfate and dextran sulfate diminished the thrombin inhibitory activity of heparin cofactor II. Other serpins, including antithrombin III and pigment epithelium-derived factor, were also degraded by heparan sulfate. This is the first evidence of acidic polysaccharides exhibiting protein-degrading activity without the aid of other proteins. PMID:25600805

  1. Bugs as drugs, part two: worms, leeches, scorpions, snails, ticks, centipedes, and spiders.

    Cherniack, E Paul


    In this second of a two-part series analyzing the evidence for the use of organisms as medicine, the use of a number of different "bugs" (worms, leeches, snails, ticks, centipedes, and spiders) is detailed. Several live organisms are used as treatments: leeches for plastic surgery and osteoarthritis and the helminths Trichuris suis and Necator americanus for inflammatory bowel disease. Leech saliva is the source of a number of anticoagulants, including the antithrombin agent hirudin and its synthetic analogues, which have been approved for human use. Predatory arthropods, such as certain species of snails, spiders, scorpions, centipedes, and ticks provide a trove of potential analgesic peptides in their venom. A synthetic analogue of a snail venom peptide, ziconotide, has been approved for human use and is used as an alternative to opioids in severe pain cases. Arthropods, such as ticks, have venom that contains anticoagulants and centipede venom has a protein that corrects abnormalities in lipid metabolism. PMID:21438646

  2. Heparin kinetics

    The author has studied the kinetics of heparin and heparin fractions after intravenous administration in humans and in this thesis the results of this study are reported. Basic knowledge about the physico-chemical properties of heparin and its interactions with proteins resulting in anticoagulant and lipolytic effects are discussed in a review (chapter II), which also comprises some clinical aspects of heparin therapy. In chapter III the kinetics of the anticoagulant effect are described after intravenous administration of five commercial heparin preparations. A mathematical model is presented that fits best to these kinetics. The kinetics of the anticoagulant and lipolytic effects after intravenous injection of various 35S-radiolabelled heparin fractions and their relationship with the disappearance of the radiolabel are described in chapter IV. Chapter V gives a description of the kinetics of two radiolabels after injection of in vitro formed complexes consisting of purified, 125I-radiolabelled antithrombin III and various 35S-radiolabelled heparin fractions. (Auth.)

  3. Chorioamnionitis caused by Serratia marcescens in a non-immunocompromised host.

    Shimizu, S; Kojima, H; Yoshida, C; Suzukawa, K; Mukai, H Y; Hasegawa, Y; Hitomi, S; Nagasawa, T


    A 26 year old pregnant woman with antithrombin III deficiency developed recurrent septicaemia with Serratia marcescens. In spite of the administration of antibiotics, high grade fever persisted. She subsequently manifested lower abdominal pain, and spontaneous abortion occurred. After the abortion, she became completely afebrile. The amnion was turbid, and microscopic examination of the placenta showed haemorrhage and massive infiltration of neutrophils, suggestive of infectious chorioamnionitis. Pulsed field gel electrophoresis showed that isolates from the blood, urine, and vaginal discharge were genetically identical. Intravenous pyelography revealed that she had a bilateral completed double ureter. It was thought that a urinary tract anomaly caused infection with S marcescens, and the pathogen spread to the chorioamnion via the bloodstream. This is the first report of chorioamnionitis caused by S marcescens in a non-immunocompromised host. In addition, these findings indicate that the chorioamnion can serve as a site for persistent infection in normal pregnancies. PMID:14600137




    Full Text Available ABSTRACT: There has been a rising incidence of myocardial infarction as a whole in recent times much because of changing pattern of life style , urbanization , changing food habits , and increase in cigarette smoking , psycho social stress and increase in the incidence of diabete s mellitus. Adding to this there is drastic increase in the incidence of MI in young people much to the influence of novel risk factors which are of current debate especially homocysteine , lipoprotein little (a , fibrinogen , anticardiolipin antibody , prote in C , protein S and antithrombin deficiency. The first ever positive prospective study on plasma homocystein and CAD was reported in 1992 by Selhubetal who showed an association of plasma homocysteine with extracranial carotid stenosis in the elderly. We r eport our study to analyze the special risk factors of MI in patients younger than 40years of age in special reference to serum homocysteine levels.

  5. Cerebral venous sinus thrombosis as presenting feature of ulcerative colitis.

    Ennaifer, R; Moussa, A; Mouelhi, L; Salem, M; Bouzaidi, S; Debbeche, R; Trabelsi, S; Najjar, T


    Thrombosis is a well recognized complication of inflammatory bowel disease that occurs in 1.3 to 6.4% of patients, however, cerebral vascular involvement is unusual. We present the case of a 16-year-old female in whom cerebral venous thrombosis was the presenting symptom of an active ulcerative pancolitis. Thrombophilia screen (plasma levels of proteins C and S, antithrombin, antibeta2-glycoprotein, lupus anticoagulant and anticardiolipin antibodies, activated protein C resistance, homocystein level antinuclear antibodies) was negative. The patient was successfully treated with anticoagulant therapy, phenobarbital and sulfasalazine. Cerebral venous thrombosis is an exceptional presenting feature of ulcerative colitis. Disease activity may play a major role in the occurrence of thrombosis. PMID:19902870

  6. Etiological analysis of presumed perinatal stroke.

    Kocaman, Canan; Yilmaz, Yuksel


    This study aimed to investigate the maternal, pre- and perinatal, and prothrombotic factors with congenital hemiparesis due to presumed perinatal stroke (PPS). Prothrombotic risk factors including protein C and S, antithrombin III, lipoprotein (a), homocystein, factor VIII levels; anticardiolipin antibodies and lupus anticoagulant; methylenetetrahydrofolate reductase mutations, factor V Leiden, prothrombin G20210A mutations were investigated. Arterial ischemic stroke was detected in 60% and periventricular venous infarction in 40%. At least one prothrombotic risk factor was present in 69%, two in 17%, and three or more in 8.5% of cases. The most common combination was methylenetetrahydrofolate reductase C677T and factor V Leiden heterozygosity. The etiology and pathogenesis of PPS is still unclear. According to this study, most of the patients with PPS might have one or more prothrombotic risk factors and certain prenatal risk factors including intrauterine growth retardation, twin gestation and preeclampsia might be related to PPS. PMID:21561729

  7. 一例接受阿那曲唑和预防性那屈肝素治疗的乳腺癌妇女发生肺动脉栓塞%Pulmonary thromboembolism in a breast cancer woman receiving anastrozole and prophylactic nadroparin


    Hormonal treatment is widely accepted for the adjuvant treatment of breast carcinoma, in order to get a reduction in the synthesis of estrogen or to block estrogen receptors in tumors that are hormone dependent. There are multiple risk factors that contribute to hypercoagulability in cancer patients. Hormonal therapy and chemotherapy are the main one. Type and stage of malignancy are other risk factors; so age, immobility and surgery are. The main antineoplastic therapy with definitive hypercoagulable effect is tamoxifen, because it can cause reduction in the concentrations of antithrombin Ⅲ and protein C. Here, we explain the case of a 75-year-old postmenopausal woman presenting with breast carcinoma who suffered from pulmonary thromboembolism during the treatment with anastrozole although she was taking nadroparin.

  8. The coagulopathy in sepsis: significance and implications for treatment

    Berardino Pollio


    Full Text Available Sepsis related coagulopathy ranges from mild laboratory alterations up to severe disseminated intravascular coagulation (DIC. There is evidence that DIC is involved in the pathogenesis of microvascular dysfunction contributing to organ failure. Additionally, the systemic activation of coagulation, by consuming platelets and coagulation factors, may cause bleeding. Thrombin generation via the tissue factor/factor VIIa route, contemporary depression of antithrombin and protein C anticoagulant system, as well as impaired fibrin degradation, due to high circulating levels of PAI-1, contribute to enhanced intravascular fibrin deposition. This deranged coagulopathy is an independent predictor of clinical outcome in patients with severe sepsis. Innovative supportive strategies aiming at the inhibition of coagulation activation comprise inhibition of tissue factor-mediated activation or restoration of physiological anticoagulant pathways, as the administration of recombinant human activated protein C or concentrate. In spite of some promising initial studies, additional trials are needed to define their clinical effectiveness in adults and children with severe sepsis.


    Dr. Sheikh Sajjadieh Mohammad Reza


    Full Text Available Patients with advanced hepatic failure may present with the entire spectrum of coagulation factor deficiencies. This study was designed to determine laboratory abnormalities in coagulation in chronic liver disease and the association of these abnormalities with the extent of chronic hepatitis and cirrhosis. Coagulation markers were assayed in 60 participants: 20 patients with chronic hepatitis, 20 patients with cirrhosis, and 20 healthy individuals (control. Plasma levels of anti-thrombin III were determined by a chromogenic substrate method, and plasma concentrations of fibrinogen were analyzed by the Rutberg method. Commercially available assays were used for laboratory coagulation tests. The levels of coagualation activity markers in patients with chronic liver disease were significantly different in comparison to those in healthy participants. These results indicate the utility of measuring markers for coagulation activity in determining which cirrhosis patients are more susceptible to disseminated intravascular coagulation.

  10. Inhibition of coagulation factors by recombinant barley serpin BSZx

    Dahl, Søren Weis; Rasmussen, S.K.; Petersen, L..C.;


    leukocytes, a fungal trypsin and three subtilisins, Thrombin, plasma kallikrein, factor VIIa/tissue factor and factor Xa were inhibited by BSZx at heparin independent association rates (k(ass)) of 4.5 x 10(3)-1.3 x 10(5) M(-1) s(-1) at 22 degrees C. Only factor Xa turned a significant fraction of BSZx over...... as substrate, Complexes of these proteinase with BSZx resisted boiling in SDS, and amino acid sequencing showed that cleavage in the reactive center loop only occurred after P-1 Arg. Activated protein C and leukocyte elastase were slowly inhibited by BSZx (k(ass) = 1-2 x 10(2) M(-1) s(-1)) whereas...... factor XIIa, urokinase and tissue type plasminogen activator, plasmin and pancreas kallikrein and elastase were not or only weakly affected, The inhibition pattern with mammalian proteinases reveal a specificity of BSZx similar to that of antithrombin III. Trypsin from Fusarium was not inhibited while...

  11. Structural characterization of pharmaceutical heparins prepared from different animal tissues.

    Fu, Li; Li, Guoyun; Yang, Bo; Onishi, Akihiro; Li, Lingyun; Sun, Peilong; Zhang, Fuming; Linhardt, Robert J


    Although most pharmaceutical heparin used today is obtained from porcine intestine, heparin has historically been prepared from bovine lung and ovine intestine. There is some regulatory concern about establishing the species origin of heparin. This concern began with the outbreak of mad cow disease in the 1990s and was exacerbated during the heparin shortage in the 2000s and the heparin contamination crisis of 2007-2008. Three heparins from porcine, ovine, and bovine were characterized through state-of-the-art carbohydrate analysis methods with a view profiling their physicochemical properties. Differences in molecular weight, monosaccharide and disaccharide composition, oligosaccharide sequence, and antithrombin III-binding affinity were observed. These data provide some insight into the variability of heparins obtained from these three species and suggest some analytical approaches that may be useful in confirming the species origin of a heparin active pharmaceutical ingredient. PMID:23526651

  12. [The correction of functional disorders of the hemostatic system and of the rheological properties of the blood in dogs in the late period of hemorrhagic shock by the intravenous transfusion of lactoprotein].

    Oborin, A N; Uspenskiĭ, B A; Kondratskiĭ, B A; Mindiuk, M V


    In 9 dogs with severe hemorrhagic shock, the effect of hemocorrector "Lactoprotein" on the indices of coagulative hemostasis and rheologic blood properties was studied. It was established that in intravenous transfusion of lactoprotein at a dose of 10 ml/kg permitting to lead the animals out from the state of shock, the syndromes of disseminated intravascular coagulation and high viscosity of the blood were cupped off. However, by the end of 48 hours of observation, fibrinogen level in the blood of the animals increased sharply, while antithrombin-III concentration and hematocrit decreased. Together with recommendation to use lactoprotein in the complex of shock therapy at all the stages of medical evacuation, the conclusion about necessity to perform at the early postshock period the differential component hemo- and anticoagulative therapy has been made. PMID:1291772

  13. [Variations in hemostasis and fibrinolysis during the treatment of acute myocardial infarct (AMI) with tissue-type plasminogen activator (TTPA). A study of 17 cases].

    Izaguirre Avila, R; Ruiz de Chávez Cervantes, A; Villavicencio, R; Gómez Trigos, A; Mar Chavira, R; Spíndola, M del C; Casanova, J M


    The aim of this trial was to estimate changes in the coagulation and fibrinolysis systems during the thrombolytic treatment with recombinant human tissue-type plasminogen activator (rt-PA) in patients with acute myocardial infarction and correlate with hemorrhagic complications. We studied 17 patients with a 3 hours-continuous systemic infusion of 100 mg of rt-PA. Prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen splits products, plasminogen, alfa-2-antiplasmin (a-2AP) and antithrombin III (AT-III) were performed before, during and after infusion. Most patients showed lengthening coagulation times. Fibrinogen and plasminogen were decreased and PDF was increased. No variations in alpha-2AP or AT-III were observed. The recuperation of fibrinogen levels occurred in 3 hours and there was hyperfibrinogenemia after day 3. No hemorrhagic complication was observed in patients with abnormalities in these coagulation or fibrinolytic tests. PMID:8347053

  14. Integrated laboratory coagulation tests in hypercoagulation diagnosis and thrombosis risk assessment. Part I. The pathophysiology of thrombosis and hypercoagulation

    E. N. Lipets


    Full Text Available Thrombosis is a fatal hemostatic disorders occurring in various conditions ranging from pregnancy and surgery to cancer, sepsis and heart attack. Despite the availability of different anticoagulants and accumulated clinical experience, proving their effectiveness, thrombosis remains a major cause of morbidity and mortality. This is largely due to the fact that conventional laboratory coagulation tests are not sufficiently sensitive to the hypercoagulable state, and they are difficult to use for assessing the risk of thrombosis. Specific molecular markers (D-dimers, fibrinopeptide, thrombin-antithrombin complex are more effective, but also have a large number of disadvantages. A possible solution is the use of integrated test, which simulate in vitro the majority of the physiological coagulation processes. In the first part of this paper the biochemical processes that cause the risk of thrombosis were discussed.

  15. Molecular evolution of serpins: homologous structure of the human α1-antichymotrypsin and α1-antitrypsin genes

    α1-Antichymotrypsin belongs to a supergene family that includes α1-antitrypsin, antithrombin III, ovalbumin, and angiotensinogen. The human chromosomal α1-antichymotrypsin gene has been cloned and its molecular structure established. The gene is approximately 12 kb in length and contains five exons and four introns. The locations of the introns within the α1-antichymotrypsin gene are identical with those of the human α1-antitrypsin and angiotensinogen genes. Other members of this supergene family contain introns located at nonhomologous positions of the genes. The homologous organization of the α1-antichymotrypsin and α1-antitrypsin genes corresponds with the high degree of homology between their protein sequences and suggest that these loci arose by recent gene duplication. A model is presented for the evolution of both the genomic structure and the protein sequences of the serine protease inhibitor superfamily

  16. Sulfated levan from Halomonas smyrnensis as a bioactive, heparin-mimetic glycan for cardiac tissue engineering applications.

    Erginer, Merve; Akcay, Ayca; Coskunkan, Binnaz; Morova, Tunc; Rende, Deniz; Bucak, Seyda; Baysal, Nihat; Ozisik, Rahmi; Eroglu, Mehmet S; Agirbasli, Mehmet; Toksoy Oner, Ebru


    Chemical derivatives of levan from Halomonas smyrnensis AAD6(T) with low, medium and high levels of sulfation were synthesized and characterized by FTIR and 2D-NMR. Sulfated levan samples were found to exhibit anticoagulation activity via the intrinsic pathway like heparin in a dose-dependent manner. Exceptionally high heparin equivalent activity of levan sulfate was shown to proceed via thrombin inhibition where decreased Factor Xa activity with increasing concentration was observed in antithrombin tests and above a certain concentration, levan sulfate showed a better inhibitor activity than heparin. In vitro experimental results were then verified in silico by docking studies using equilibrium structures obtained by molecular dynamic simulations and results suggested a sulfation dependent binding mechanism. With its high biocompatibility and heparin mimetic activity, levan sulfate can be considered as a suitable functional biomaterial to design biologically active, functionalized, thin films and engineered smart scaffolds for cardiac tissue engineering applications. PMID:27261753

  17. [Anticoagulant activity of low-molecular-weight sulfated derivatives of galactomannan from Cyamopsis tetragonoloba (L.) seeds].

    Mestechkina, N M; Shcherbukhin, V D; Bannikova, G E; Varlamov, V P; Drozd, N N; Tolstenkov, A S; Makarov, V A; Tikhonov, V E


    Galactomannan from seeds of Cyamopsis tetragonoloba (L.) Taub. (guar) was depolymerized using immobilized enzymatic preparation celloviridin. A set of fragments whose molecular weights varied from 12.6 to 245.6 kDa was obtained. Sulfated derivatives of components of all fractions were synthesized, in which the content of HSO3(-) groups was 48.05% +/- 2.31. All preparations exhibited anticoagulant activity, which was recorded in vitro in two tests--aIIa and aXa. The antithrombin activity (aIIa) was high (up to 65-87 U/mg) and did not depend on the molecular weight of a sulfated derivative; in the second test (aXa), the effect of molecular weight was observed. Biospecific electrophoresis allowed us to detect the ability of galactomannan sulfates to form complexes with protamine sulfate, a classic antidote to heparin. PMID:18491607

  18. Hemostatic response to surgical neutering via ovariectomy and ovariohysterectomy in dogs

    Moldal, Elena R.; Kristensen, Annemarie Thuri; Peeters, Marijke E.;


    Objective-To investigate the hemostatic response to surgery and compare the response for ovariohysterectomy with that for ovariectomy and to evaluate the usefulness of thromboelastography on plasma samples. Animals-42 female dogs. Procedures-Dogs were assigned to undergo ovariohysterectomy or...... antigen, fibrinogen, antithrombin, and protein C; activity of factor VIII; activated partial thromboplastin time; prothrombin time; and thrombin time. The fibrinolytic response was assessed via concentrations of D-dimer, plasminogen, and α-2-antiplasmin (plasmin inhibitor). Results-Substantial hemostatic...... prothrombin and thrombin times. The dogs also typically had activation of the fibrinolytic system, as evidenced by increased postoperative concentrations of D-dimer, plasminogen, and plasmin inhibitor. Differences between the 2 groups could not be detected for any variables. Conclusions and Clinical Relevance...

  19. TACTIC: Trans-Agency Consortium for Trauma-Induced Coagulopathy.

    Mann, K G; Freeman, K


    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. PMID:26149052

  20. Stent thrombosis and restenosis: what have we learned and where are we going? The Andreas Grüntzig Lecture ESC 2014.

    Byrne, Robert A; Joner, Michael; Kastrati, Adnan


    Modern-day stenting procedures leverage advances in pharmacotherapy and device innovation. Patients treated with contemporary antiplatelet agents, peri-procedural antithrombin therapy and new-generation drug-eluting stents (DES) have excellent outcomes over the short to medium term. Indeed, coupled with the reducing costs of these devices in most countries there remain very few indications where patients should be denied treatment with standard-of-care DES therapy. The two major causes of stent failure are stent thrombosis (ST) and in-stent restenosis (ISR). The incidence of both has reduced considerably in recent years. Current clinical registries and randomized trials with broad inclusion criteria show rates of ST at or strategies which circumvent the need for chronically indwelling stents--such as drug-coated balloons or fully bioresorbable stents-more data are needed before the wider use of these therapies can be advocated. PMID:26417060

  1. Inherited thrombophilia and reproductive disorders

    Liatsikos, Spyros A.; Tsikouras, Panagiotis; Manav, Bachar; Csorba, Roland; von Tempelhoff, Georg Friedrich; Galazios, Georgios


    Apart from its established role in the pathogenesis of venous thromboembolism (VTE), inherited thrombophilia has been proposed as a possible cause of pregnancy loss and vascular gestational complications. There is a lot of controversy in the literature on the relationship between inherited prothrombotic defects and these obstetric complications. This is a review of the literature on inherited thrombophilia and reproductive disorders. Factor V Leiden, prothrombin G20210A mutation, and protein S deficiency seem to be associated with late and recurrent early pregnancy loss, while their impact on other pregnancy complications is conflicting. No definite association has been established between protein C and antithrombin deficiency and adverse pregnancy outcome, primarily due to their low prevalence. Screening is suggested only for women with early recurrent loss or late pregnancy loss. Anticoagulant treatment during pregnancy should be considered for women with complications who were tested positive for thrombophilia. PMID:27026779

  2. Predictors of fatal outcomes resulting from acute Escherichia coli mastitis in dairy cows.

    Hagiwara, Seiichi; Mori, Kouichiro; Nagahata, Hajime


    To evaluate the prognostic criteria for identifying cows at an increased risk of a fatal outcome from acute Escherichia coli mastitis, the potential cut-off values for five diagnostic parameters associated with a high mortality were determined by receiver operator characteristic curve analysis. These criteria were hematocrit value >32%, blood non-esterified fatty acid concentration >0.4 mEq/l, antithrombin activity <120%, platelet count <15 × 10(4)/ml and presence of dysstasia. Exceeding the cut-off values for at least three parameters on day 2 after onset predicted fatality (predictive value 87.5). When these prognostic criteria were applied to 34 clinical cases, cows that met three criteria were seven times more likely to die than cows that met fewer than three criteria. PMID:26875836

  3. Alterations in coagulation following major liver resection.

    Mallett, S V; Sugavanam, A; Krzanicki, D A; Patel, S; Broomhead, R H; Davidson, B R; Riddell, A; Gatt, A; Chowdary, P


    The international normalised ratio is frequently raised in patients who have undergone major liver resection, and is assumed to represent a potential bleeding risk. However, these patients have an increased risk of venous thromboembolic events, despite conventional coagulation tests indicating hypocoagulability. This prospective, observational study of patients undergoing major hepatic resection analysed the serial changes in coagulation in the early postoperative period. Thrombin generation parameters and viscoelastic tests of coagulation (thromboelastometry) remained within normal ranges throughout the study period. Levels of the procoagulant factors II, V, VII and X initially fell, but V and X returned to or exceeded normal range by postoperative day five. Levels of factor VIII and Von Willebrand factor were significantly elevated from postoperative day one (p < 0.01). Levels of the anticoagulants, protein C and antithrombin remained significantly depressed on postoperative day five (p = 0.01). Overall, the imbalance between pro- and anticoagulant factors suggested a prothrombotic environment in the early postoperative period. PMID:27030945

  4. Modeling the microscopic electrical properties of thrombin binding aptamer (TBA) for label-free biosensors

    Alfinito, Eleonora; Cataldo, Rosella; De Nunzio, Giorgio; Giotta, Livia; Guascito, Maria Rachele


    Aptamers are chemically produced oligonucleotides, able to bind a variety of targets such as drugs, proteins and pathogens with high sensitivity and selectivity. Therefore, aptamers are largely employed for producing label-free biosensors, with significant applications in diagnostics and drug delivery. In particular, the anti-thrombin aptamers are biomolecules of high interest for clinical use, because of their ability to recognize and bind the thrombin enzyme. Among them, the DNA 15-mer thrombin-binding aptamer (TBA), has been widely explored concerning both its structure, which was resolved with different techniques, and its function, especially about the possibility of using it as the active part of biosensors. This paper proposes a microscopic model of the electrical properties of TBA and the aptamer-thrombin complex, combining information from both structure and function. The novelty consists in describing both the aptamer alone and the complex as an impedance network, thus going deeper inside the issues...

  5. Heparin based polyurethanes: A state-of-the-art review.

    Zia, Fatima; Zia, Khalid Mahmood; Zuber, Mohammad; Tabasum, Shazia; Rehman, Saima


    Polyurethanes (PUs) are considered currently as one of the established bio compatible and blood compatible biomaterials offering tremendous structure-property relationship. But few limitations such as low resistance to micro-emboli and thrombi are still associated with these biomaterials that restricted their applications and hence need to be modified. Heparin, a highly sulfonated and negatively charged member of glycosaminoglycan family is well established for their anti-thrombin, anticoagulant and many biological activities that make it a highly attractive candidate capable of modifying or tailoring polymer properties. Incorporation of heparin for the improvement of biocompatibility of PUs is an interesting approach and enabling emerging technology. This review focuses on the methods used for modification of PUs via heparin with their pros and cons. The major PU-heparin systems with the recent developments and their possible biomedical applications are discussed. PMID:26666430

  6. Cerebral venous thrombosis in adults: A study of 50 cases from Iran

    Lotfi J


    Full Text Available The study was conducted to determine the clinical pattern and etiology of cerebral venous thrombosis in Iran. Records of all adult patients admitted with a clinical diagnosis and MRI of cerebral venous thrombosis from 1993 through 1999 in three major hospitals of Tehran, Iran were reviewed. Fifty patients (39 women, 11 men aged 14 to 56 years were identified. Headache the most frequent and often the earliest symptom, was encountered in 82 percent and papilledema, the most frequent sign in 56 percent of cases. Infection was the cause of cerebral venous thrombosis in 26 percent of our cases. Other causes included oral contraceptive (32 percent, vasculitis (6 percent, Behcet's disease (4 percent, postpartum state (4 percent, myeloproliferative disease (2 percent, ulcerative colitis (2 percent, antithrombin III deficiency (2 percent and diabetic ketoacidosis (2 percent. Oral contraceptive was recognized as the most common etiology. Infection is an important cause whereas procoagulation disorders are uncommon.

  7. 肿瘤性DIC实验室指标诊断阳性率分析%Analysis of Laboratory Index Positive Diagnostic Rate of Malignancy Tumor with Disseminated Intravascular Coagulation



    目的探讨恶性肿瘤合并弥散性血管内凝血(Dis eminated Intravascular Coagulation,DIC)时,实验室检测指标DIC三项咱D-二聚体(D-dimer,DD)、纤维蛋白原降解产物(Fibrinogen Degradation Product,FDP)、抗凝血酶原II(AntithrombinII,ATII)暂和凝血常规咱凝血酶原时间(Prothrombin Time,PT)、活化部分凝血酶原时间(Activated Partial Prothrombin Time,APTT)、纤维蛋白原(Fibrinogen,FIB)检测阳性率的情况,并比较各组之间的有效性。方法回顾性分析78例恶性肿瘤患者的临床及实验室资料。结果 DD是恶性肿瘤合并DIC时诊断阳性率最高的指标。结论在肿瘤合并DIC时,DD的诊断价值最有意义。%Objective To explore malignancy tumor with disseminated intravascular coagulation,which laboratory index of D-dimer、fibrinogen degradation product、AntithrombinI I、prothrombin time、activated partial prothrombin time、fibrinogen are the positive indicator.Methods Clinical and laboratory data for 78 cases of DIC with malignancy tumor were retrospectively analysed.Result:D-dimer is the most ef ective indicator of DIC with malignancy tumor.Results The diagnosis index of:DD is the highest positive rate of malignant tumor complicated with DIC. Conclusion In the tumor with DIC, the diagnostic value of DD was the most significant.

  8. Systemic coagulation parameters in mice after treatment with vascular targeting agents

    Gottstein Claudia


    Full Text Available Abstract Background Vascular targeting of malignant tumors has become a clinically validated new treatment approach with clear patient benefit. However clinical studies have also revealed that some types of vascular targeting agents (VTAs are prone to coagulation system side effects. It is therefore essential to predetermine coagulation parameters in preclinical studies. As of to date, this has rarely been done, predominantly due to technical issues. The goal of this study was to establish and apply a standardized process, whereby systemic coagulation activation can be routinely measured in mice. Results We have evaluated a number of sampling techniques and coagulation tests regarding their suitability for this purpose. We were able to adapt two assays measuring soluble fibrin, a marker for a prethrombotic status. Thus, soluble fibrin could be measured for the first time in mice. All assays were validated in a positive control model for systemic coagulation activation, i.e. lipopolysaccharide-induced endotoxemia. Based on our results, we selected a panel of coagulation tests, which are both feasable and informative for preclinical testing of VTAs: soluble fibrin, thrombin-antithrombin complexes, free antithrombin III, white blood cell counts and platelet counts. The effect of tumor transplants on coagulation parameters was evaluated using this panel. We then applied this set of assays in treatment studies with a VTA developed in our laboratory to investigate a potential systemic coagulation activation. Conclusion We have established a standardized panel of assays that can be used to test murine blood samples for coagulation activation in preclinical studies. All tests are feasible to perform in any research laboratory without specialized equipment. In addition, this is the first report to measure soluble fibrin, an early marker of systemic coagulation activation, in mice. The panel was applied on tumor bearing mice and mice treated with a VTA

  9. Circulating Thrombotic Risk Factors in Young Patients with Coronary Artery Disease Who Are on Statins and Anti-platelet Drugs.

    George, Reema; Sivadasanpillai, Harikrishnan; Jayakumari, Narayani; Bhatt, Anugya; Thulaseedharan, Jissa V; Tharakan, Jaganmohan A


    Thrombotic risk factors may contribute to premature coronary artery disease (CAD), in addition to the conventional risk factors. There is paucity of data on studies evaluating the role of thrombotic factors in premature CAD in Indian patients. Thus a case-control study was performed to evaluate the role of thrombotic and atherogenic factors in young patients with angiographically proven CAD who are on treatment with statins and anti-platelet drugs. 152 patients (≤55 years) with angiographically proven CAD and 102 asymptomatic controls were recruited. Clinical and biochemical data were obtained in both groups. Blood levels of thrombotic factors-fibrinogen, antithrombin-III, tissue-plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), von-Willebrand factor (v-WF), lipoprotein(a) [Lp(a)] and homocysteine were analyzed. Patients had high levels of conventional CAD risk factors (diabetes mellitus, smoking, hypertension, dyslipidemia and positive family history) compared to controls. Logistic regression analysis revealed that low antithrombin-III (odds ratio/OR 11.2; 95 % confidence interval/CI 2.29-54.01), high fibrinogen (OR 6.04; 95 % CI 1.09-33.21) and high Lp(a) (OR 4.54; 95 % CI 0.92-22.56), as important, independent risk factors in patients. PAI-1(OR 0.15; 95 % CI 0.03-0.69) levels were significantly lower in patients. But other thrombotic risk factors studied (t-PA, v-WF and homocysteine) were comparable among patients and controls. The treatment using statins and anti-platelet drugs might be contributing to the control of some of the thrombotic risk factors. The strategies aiming at lowering the levels of thrombotic risk factors along with conventional risk factors may be useful in primary and secondary prevention of CAD. PMID:27382201

  10. Deciphering the main venom components of the ectoparasitic ant-like bethylid wasp, Scleroderma guani.

    Zhu, Jia-Ying


    Similar to venom found in most venomous animals, parasitoid venoms contain a complex cocktail of proteins with potential agrichemical and pharmaceutical use. Even though parasitoids are one of the largest group of venomous animals, little is known about their venom composition. Recent few studies revealed high variated venom composition existing not only in different species but also between closely related strains, impling that increasing information on the venom proteins from more greater diversity of species of different taxa is key to comprehensively uncover the complete picture of parasitoid venom. Here, we explored the major protein components of the venom of ectoparasitic ant-like bethylid wasp, Scleroderma guani by an integrative transcriptomic-proteomic approach. Illumina deep sequencing of venom apparatus cDNA produced 49,873 transcripts. By mapping the peptide spectral data derived from venom reservoir against these transcripts, mass spectrometry analysis revealed ten main venom proteins, including serine proteinase, metalloprotease, dipeptidyl peptidase IV, esterase, antithrombin-III, acid phosphatase, neural/ectodermal development factor IMP-L2 like protein, venom allergen 3, and unknown protein. Interestingly, one serine proteinase was firstly identified with rarely high molecular weight about 200 kDa in parasitoid venom. The occurrence of abundant acid phosphatase, antithrombin-III and venom allergen 3 demonstrated that S. guani venom composition is similar to that of social wasp venoms. All identified venom genes showed abundantly biased expression in venom apparatus, indicating their virulent functions involved in parasitization. This study shed light on the more better understanding of parasitoid venom evolution across species and will facilitate the further elucidation of function and toxicity of these venom proteins. PMID:26853496

  11. Evaluation of selected thrombotic factors among pregnant women with preeclampsia and normal pregnant women

    Saghafi, Nafiseh; Mohammadzadeh Vatanchi, Atieh; Tara, Fatemeh; Pourali, Leila; Dadgar, Salmeh


    Background: Preeclampsia is one of the common complications during pregnancy with considerable maternal and fetal mortality and morbidity. Hypercoagulability due to thrombophilic factors is discussed as the etiology involved in this disease. Objective: The aim of this study was to evaluate selected thrombotic factors among pregnant women with preeclampsia and normal pregnant women. Materials and Methods: This case-control study was performed on 200 pregnant women at third trimester of pregnancy between 2012 and 2013. 100 pregnant women admitted to Qaem and Imam Reza hospitals of Mashhad, due to preeclampsia, were selected as case group and 100 pregnant women without preeclampsia referred to OB/GYN clinic of these hospitals as control group. Blood samples were taken from two groups for evaluation of the coagulation factors including factor V Leiden, protein C, protein S, antithrombin III, anti-cardiolipin antibodies, and lupus anticoagulant antibodies. Results: Two groups were not significantly different in terms of maternal age and parity (p>0.05). Levels of factor V Leiden, protein C, protein S, antithrombin III, anti-cardiolipin antibodies and lupus anticoagulant antibodies were compared between two groups. The number of patients with abnormal factor V Leiden and protein C was significantly higher in case group than in the control group (p<0.01 respectively), but other factors were not significant different between two groups. Thrombophilia disorders were significantly more in case group compared to control (p<0.001). Conclusion: The risk of thrombophilia disorders is higher in preeclamptic patients than normal pregnant women. PMID:25709635

  12. Expression and Characterization of Gly-317 Variants of Factor IX Causing Variable Bleeding in Hemophilia B Patients.

    Lu, Qiuya; Yang, Likui; Manithody, Chandrashekhara; Wang, Xuefeng; Rezaie, Alireza R


    We recently identified two hemophilia B patients who carried Gly-317 to Arg (FIX-G317R) or Gly-317 to Glu (FIX-G317E) substitutions in their FIX gene. The former mutation caused severe and the latter moderate bleeding in afflicted patients. To understand the molecular basis for the variable clinical manifestation of Gly-317 mutations, we prepared recombinant G317R and G317E derivatives of FIX and compared their kinetic properties to those of recombinant wild-type FIX in appropriate assay systems. Both physiological activators, factor XIa and extrinsic Tenase (factor VIIa-tissue factor), activated both zymogen variants with an ∼1.5-fold elevated K(m); however, extrinsic Tenase activated FIX-G317E with an ∼2-fold improved k(cat). By contrast to zymogen activation, the catalytic activities of both FIXa-G317R and FIXa-G317E enzymes toward the natural substrate, factor X, were dramatically (>4 orders of magnitude) impaired, but their apparent affinity for interaction with factor VIIIa was only slightly (<2-fold) decreased. Further studies revealed that the reactivity of FIXa-G317R and FIXa-G317E with antithrombin has been impaired 10- and 13-fold, respectively, in the absence and 166- and 500-fold, respectively, in the presence of pentasaccharide. As expected, the clotting activities of FIX variants could not be measured by the aPTT assay. These results implicate a critical role for Gly-317 in maintaining normal catalytic function for FIX/FIXa in the clotting cascade. The results further suggest that improved k(cat) of FIX-G317E activation in the extrinsic pathway together with dramatically impaired reactivity of FIXa-G317E with antithrombin may account for the less severe bleeding phenotype of a hemophilia B patient carrying the FIX-G317E mutation. PMID:26023895

  13. Inactivation of factor XII active fragment in normal plasma. Predominant role of C-1-inhibitor.

    de Agostini, A; Lijnen, H R; Pixley, R A; Colman, R W; Schapira, M


    To define the factors responsible for the inactivation of the active fragment derived from Factor XII (Factor XIIf ) in plasma, we studied the inactivation kinetics of Factor XIIf in various purified and plasma mixtures. We also analyzed the formation of 125I-Factor XIIf -inhibitor complexes by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In purified systems, the bimolecular rate constants for the reactions of Factor XIIf with C-1-inhibitor, alpha 2-antiplasmin, and antithrombin III were 18.5, 0.91, and 0.32 X 10(4) M-1 min-1, respectively. Furthermore, SDS-PAGE analysis revealed that 1:1 stoichiometric complexes were formed between 125I-Factor XIIf and each of these three inhibitors. In contrast, kinetic and SDS-PAGE studies indicated that Factor XIIf did not react with alpha 1-antitrypsin or alpha 2-macroglobulin. The inactivation rate constant of Factor XIIf by prekallikrein-deficient plasma was 14.4 X 10(-2) min-1, a value that was essentially identical to the value predicted from the studies in purified systems (15.5 X 10(-2) min-1). This constant was reduced to 1.8 X 10(-2) min-1 when Factor XIIf was inactivated by prekallikrein-deficient plasma that had been immunodepleted (less than 5%) of C-1-inhibitor. In addition, after inactivation in normal plasma, 74% of the active 125I-Factor XIIf was found to form a complex with C-1-inhibitor, whereas 26% of the enzyme formed complexes with alpha 2-antiplasmin and antithrombin III. Furthermore, 42% of the labeled enzyme was still complexed with C-1-inhibitor when 125I-Factor XII was inactivated in hereditary angioedema plasma that contained 32% of functional C-1-inhibitor. This study quantitatively demonstrates the dominant role of C-1-inhibitor in the inactivation of Factor XIIf in the plasma milieu. PMID:6725552

  14. Contraception and Thrombophilia - A statement from the German Society of Gynecological Endocrinology and Reproductive Medicine (DGGEF e. V. and the Professional Association of the German Gynaecologists (BVF e. V.

    Rabe T


    Full Text Available Venous thromboembolism (VTE is responsible for more than half a million deaths annually in the European Union, most in older people following surgery, but some in women of reproductive age using various hormonal contraceptives. In some parts of the population inherited defects of the blood coagulation system (factor V Leiden, prothrombin G20210A, protein C, protein S and antithrombin deficiency are responsible for an increased risk of VTE, which is also influenced by concomitant factors: e.g. long-distance travel, immobilisation, advanced age, cigarette smoking, high BMI, surgery, malignancy, fluid loss, pregnancy, oral contraceptive use and hormone replacement therapy (HRT. Laboratory testing: General screening for thrombophilia prior to the prescription of oral contraceptives (OC is not recommended. Laboratory testing for thrombophilia should be limited to women with a positive family and/or personal history of VTE or vascular occlusion. – Factor V Leiden is by far the most common congenital thrombophilia. Heterozygous factor V Leiden (5-fold increased VTE risk is present in 3–13%, homozygous factor V Leiden (10-fold increased VTE risk in up to 0.2–1% of people of European origin. – Prothrombin mutation G20210A: Autosomal dominant mutation inheritance (2% of people of European origin leads to a 3-fold increase in VTE risk is substantially increased if one or more additional risk factors are present such as factor V Leiden or protein C, S, or antithrombin deficiency. – Protein C and protein S: VTE risk increases with protein C or S deficiency (odds-ratio 3–15 and 5–11, respectively. – Antithrombin deficiency leads to a 4 to 50-fold increase in VTE risk depending on the type of deficiency. Female hormonal contraceptives containing progestogens with or without combination with a synthetic estrogens (mainly ethinylestradiol [EE] or a natural estrogen (e.g. estradiol or its derivative estradiol valerate affect the incidence of VTE in

  15. 体积排阻色谱法测定低分子量肝素抗凝血因子Xa的活性%An assay for anti-factor Xa activity of low molecular weight heparins by high performance liquid size exclusion chromatography

    张倩倩; 康经武


    发展了一种基于体积排阻色谱测定低分子量肝素(LMWH)抗凝血活性的方法.利用肝素与抗凝血酶Ⅲ(ATⅢ)结合后可增强ATⅢ对凝血因子Xa (FXa)抑制作用的原理,通过测定加入LMWH后FXa水解其生色底物产生对硝基苯胺(pNA)这一反应的抑制程度确定LMWH的活性.首先将含有一定浓度LMWH的缓冲溶液与ATⅢ溶液混合,然后依次加入FXa和生色底物,分别孵育一段时间.底物被FXa水解,产生游离的pNA.体积排阻色谱可将小分子产物pNA与其他大分子分离开,因而可以在pNA的最大吸收波长下得到高灵敏度的测定,并且不再受其他成分的干扰.该方法重复性好,灵敏度高,极大地减少了样品的消耗量,降低了成本,并且还可进行各种复杂样品(如血浆)中LMWH抗FXa活性的监测.%The "gold standard" assay for monitoring low molecular weight heparins (LMWHs) activity is the chromogenic-based anti-factor Xa assay.The methodology of an anti-factor Xa assay is that LMWH is added to a known amount of excess factor Xa and excess antithrombin.It will bind to antithrombin and form a triplet complex with factor Xa,inhibiting the activity of factor Xa.However,the residual factor Xa can still hydrolyze chromogenic peptide substrate,releasing the chromophore for photometric detection.The absorbance is inversely proportional to the amount of heparin/LMWH.The results are given in anticoagulant concentration in units/ mL of anti-factor Xa,such that high values indicate high levels of anticoagulation and low values indicate low levels of anticoagulation.Herein,a novel assay method for anti-FXa activity of LMWHs using high performance liquid size exclusion chromatography (SEC) is reported,in which antithrombin m (AT m) was diluted by the buffer solution contained LMWHs.Subsequently,exogenous FXa and p-nitroaniline coupled peptide substrate were added and incubated for a period,separately.The resulting mixture was separated based on size by SEC

  16. Análise da freqüência de trombofilia em pacientes com atrofia branca de Milian Frequency analysis of thrombophilia in patients with atrophie blanche

    Aline Donati Jorge


    Full Text Available FUNDAMENTOS - Atrofia branca de Milian ou vasculopatia livedóide é entidade clinicopatológica rara, cuja patogênese não é completamente compreendida. OBJETIVOS - Avaliar casos de atrofia branca de Milian para verificar a prevalência de diversas trombofilias. MATERIAL E MÉTODOS - Quatorze pacientes foram submetidos a exames laboratoriais incluindo pesquisa de fator V (Leiden, protrombina mutante, dosagem de antitrombina, proteína S e C, pesquisa de anticorpos anticardiolipina e anticoagulante lúpico, dosagem de homocisteína e pesquisa da mutação da metilenotetraidrofolatoredutase. RESULTADOS - Dos nove doentes cujos critérios de inclusão foram preenchidos para análise da freqüência de trombofilia, foram encontrados quatro com fatores relacionados à trombofilia: deficiência da antitrombina (um caso, deficiência da proteína S (um caso, mutação da metilenotetraidrofolatoredutase com hiperhomocisteinemia (um caso e presença de anticorpo anticardiolipina (um caso. CONCLUSÃO - Apesar de este estudo não apresentar casuística que possibilite a comparação com a população geral, os dados sugerem a presença de eventos geradores de trombofilia nesses doentes, contribuindo para adoção sistemática de um protocolo de investigação de trombofilia nos doentes portadores de vasculopatia livedóide no Brasil.INTRODUCTION: Atrophie blanche, or livedoid vasculopathy, is a rare clinicopathological entity of unknown etiology. A "thrombo-occlusive process" theory has recently been accepted. OBJECTIVES: To search the presence of several thrombophilic abnormalities in patients with livedoid vasculopathy. METHODS: Fourteen patients were evaluated and tested for factor V Leiden, prothrombin 20210G/A variant, antithrombin, C and S proteins, anticardiolipin and lupus anticoagulant antibodies, homocysteine and methylenetetrahydrofolate reductase mutation. RESULTS: Nine patients met all criteria to be included in the analysis and four of

  17. Quality of therapeutic plasma-requirements for marketing authorization.

    Heiden, Margarethe; Seitz, Rainer


    Fresh frozen plasma (FFP) contains higher levels of intact coagulation factors and coagulation and fibrinolysis inhibitors than solvent/detergent-treated plasma (SD plasma), and also greater residual cell contamination. SD plasma is a particle-free plasma of uniform quality. SD treatment, however, has the specific result of reducing the activities of some inhibitors. Both plasma types carry a minimal residual risk of transmitting human immunodeficiency virus (HIV)-1/2, hepatitis virus B (HBV), and hepatitis virus C (HCV), but SDP is, in addition, also safe with respect to other lipid-enveloped viruses and perhaps with respect to hepatitis virus A (HAV), also due to its antibody (Ab) content. Future revisions of therapeutic plasma safety and quality standards should consider the following points:For FFP:reduce residual cell count in all FFP units to values below 5 x 10(6) leukocytes/l;screen donors for Parvovirus B19 genome and antibodies in order to establish a sufficiently large collection of genome-negative and antibody-positive donors whose FFP can be used for selected patients;For SDP:introduce pool testing for Parvovirus B19 genome; fix an upper limit for genome and a lower limit for antibody content;in addition to the standard quality control methods for therapeutic plasma, focus on assays to test for functionally intact proteinase inhibitors such as alpha(2)antiplasmin (alpha(2)AP) and alpha(1)proteinase inhibitor (alpha(1)PI) that are important for plasma indications. Commercially available kits may not be sufficient to show changes in inhibition kinetics. For both types:introduce an activation marker such as thrombin-antithrombin complex (TAT) as a random test to monitor activation processes during withdrawal, separation, manufacturing, and storage;abolish inappropriate parameters like Antithrombin III (AT III) and coagulation factor XI that are not relevant for changes in plasma quality;finally, support every effort towards establishing an efficient

  18. Efficacy L-Arginine In Patients With Nonalcoholic Steatohepatitis Associated With Metabolic Syndrome

    Oleksandr Fediv


    Full Text Available Abstract Background and Purpose Recent research in the field of hematology indicate that among the many pathogenic mechanisms of development and progression of nonalcoholic steatohepatitis NASH which occurs on the background of the metabolic syndrome an important role is played by endothelial dysfunction and violations of haemocoagulation. The aim of this research was to study the effectiveness of L-arginine as it corrects endothelial dysfunction and disorders of homeostasis haemocoagulation link in patients with NASH associated with the metabolic syndrome. Subjects and Methods 128 patients with nonalcoholic steatohepatitis associated with metabolic syndrome were examined. Some patients 63 persons received standard treatment according to national guidelines. To another group 65 patients on the background of basic therapy L-arginine hydrochloride followed by transition to oral form of L-arginine aspartate was administered. Blood levels of stable nitrogen monoxide metabolites nitrites nitrates endothelin-1 and plasma recalcification time prothrombin time thrombin time activated partial thromboplastin time fibrinogen plasma level activity of antithrombin III and coagulation factor XIII potential activity of plasminogen plasma fibrinolytic blood activity were studied. Results Originally significantly increased levels of endothelin-1 decreased after the therapy in all studied groups but more noticeable changes in the group with L-arginine appointment were observed p0.05. In the studied groups normalization of stable nitrogen monoxide metabolites after treatment was also noticed. Significant p0.05 increase in all haemocoagulation time characteristics and activities of antithrombin-III and factor XIII was found. The positive effect of L-arginine on blood fibrinolytic activity was noted. Discussion and Conclusion Combined therapy of nonalcoholic steatohepatitis associated with metabolic syndrome with a differentiated degreeal L-arginine assignment by

  19. Activation of factor VII bound to tissue factor: A key early step in the tissue factor pathway of blood coagulation

    Whether the factor VII/tissue factor complex that forms in tissue factor-dependent blood coagulation must be activated to factor VIIa/tissue factor before it can activate its substrates, factor X and IX, has been a difficult question to answer because the substrates, once activated, back-activate factor VII. The earlier studies suggested that human factor VII/tissue factor cannot activate factor IX. Studies have now been extended to the activation of factor X. Reaction mixtures were made with purified factor VII, X, and tissue factor; in some experiments antithrombin III and heparin were added to prevent back-activation of factor VII. Factor X was activated at similar rates in reaction mixtures containing either VII or factor VIIa after an initial 30-sec lag with factor VII. In reaction mixtures with factor VII a linear activation of factor X was established several minutes before cleavage of 125I-labeled factor VII to the two-chain activated molecule was demonstrable on gel profiles. These data suggest that factor VII/tissue factor cannot activate measurable amounts of factor X over several minutes. Overall, the results support the hypothesis that a rapid preferential activation of factor VII bound to tissue factor by trace amounts of factor Xa is a key early step in tissue factor-dependent blood coagulation

  20. A study of some aspects of auto immunity in children with thalassemia major

    This study is subjected to estimate the study of the most common autoimmune complications in thalassemia and the prevalence of different auto antibodies so as to evaluate its correlation with personal data including age, sex, growth parameters, Hepatitis C virus infection ferritin level and type of iron chelation therapy methodology : Different Abs including anti cardiolipin, antitistone, ANA, Anti erythropoietin by ELISA technique Hepatitis C virus by ELISA. Ferritin by IRMA (Immune radiometric assay), Protein C, S. Antithrombin III by ELISA result of the study were analyzed by appropriate statistical methods .The results showed that age of Hepatitis C virus negative cases was significantly lower than Hepatitis C virus Positive cases, ferritin levels were significantly higher in patients on defripone than those as desferrioxamine, anti erythropoietin was detected in 93.3% of cases regarding the antithrombotic factors deficiency. Protein C was deficient in 26.67% of cases. Protein S was deficient in 13.33% only and no cases showed anti thrombin III deficiency. No special relation was found between these abnormalities and type of chelation , Conclusions: Auto immunity may be a cofactor in the development of life threatening complication

  1. Fumonisin mycotoxicosis in broilers: plasma proteins and coagulation modifications.

    Espada, Y; Ruiz de Gopegui, R; Cuadradas, C; Cabañes, F J


    The effects of fumonisin B1 (FB1) intoxication in chickens were evaluated in three experiments. Two-day-old broiler chicks were fed a diet containing 10 mg pure FB1/kg feed for 6 days; some chicks were necropsied at this time, and others were allowed to recover for 5 wk before necropsy. In two other experiments, 2-day-old chicks were fed a broiler starter ration prepared with Fusarium moniliforme culture material containing FB1; one group received 30 mg/kg for 2 wk, and another received 300 mg FB1/kg for 8 days. Compared with controls, intoxicated chicks exhibited decreased prothrombin time, increased plasma fibrinogen (not included for the group receiving 30 mg/kg of culture material), and increased antithrombin III activity. Simultaneously decreased serum albumin concentration and increased serum globulins could be observed in groups intoxicated with F. moniliforme culture material containing FB1. The group allowed to recover for 5 wk did not exhibit modifications in hemostasis or serum proteins compared with controls. The results indicate that low doses of pure FB1 (10 mg/kg) and FB1 from F. moniliforme culture material (30 mg/kg) may alter hemostasis and serum proteins in young chicks. PMID:9087322

  2. Structure and biological activity of a fucosylated chondroitin sulfate from the sea cucumber Cucumaria japonica.

    Ustyuzhanina, Nadezhda E; Bilan, Maria I; Dmitrenok, Andrey S; Shashkov, Alexander S; Kusaykin, Mikhail I; Stonik, Valentin A; Nifantiev, Nikolay E; Usov, Anatolii I


    A fucosylated chondroitin sulfate (FCS) was isolated from the body wall of Pacific sea cucumber Cucumaria japonicaby extraction in the presence of papain followed by Cetavlon precipitation and anion-exchange chromatography. FCS was shown to contain D-GalNAc, D-GlcA, L-Fuc and sulfate in molar proportions of about 1:1:1:4.5. Structure of FCS was elucidated using NMR spectroscopy and methylation analysis of the native polysaccharide and products of its desulfation and carboxyl reduction. The polysaccharide was shown to contain a typical chondroitin core → 3)-β-D-GalNAc-(1 → 4)-β-D-GlcA-(1 →. Sulfate groups in this core occupy O-4 and the majority of O-6 of GalNAc. Fucosyl branches are represented by 3,4- and 2,4-disulfated units in a ratio of 4:1 and are linked to O-3 of GlcA. In addition, ∼ 33% of GlcA are 3-O-sulfated, and hence, the presence of short fucooligosaccharide chains side by side with monofucosyl branches cannot be excluded. FCS was shown to inhibit platelets aggregation in vitro mediated by collagen and ristocetin, but not adenosine diphosphate, and demonstrated significant anticoagulant activity, which is connected with its ability to enhance inhibition of thrombin and factor Xa by antithrombin III, as well as to influence von Willebrand factor activity. The latest property significantly distinguished FCS from low-molecular-weight heparin. PMID:26681734

  3. Anticoagulant, antiplatelet and antianemic effects of Punica granatum (pomegranate) juice in rabbits.

    Riaz, Azra; Khan, Rafeeq A


    Pomegranate (Punica granatum L., Punicaceae) is a good source of minerals and phytochemicals with diverse pharmacological activities such as anxiolytic, antidepressant, hypoglycemic, hypolipidemic, and anti-inflammatory activities. Effects of P. granatum on blood parameters and coagulation have, however, been little studied. The aim of the study was to assess the outcome of P. granatum on coagulation and anticoagulation factors at different doses on blood samples of healthy white rabbits. Blood samples of the animals were collected twice during the study and biochemical assays were performed to assess the effect on hematological, coagulation, anticoagulation, and platelet aggregation. Significant changes were observed in erythrocytes, hemoglobin, and mean corpuscular hemoglobin concentration, while bleeding and thrombin time were also prolonged significantly. There was significant increase in protein C, thrombin antithrombin complex levels, and decrease in platelet aggregation and fibrinogen concentration, in a dose-dependent manner. The results of hematological and coagulation assays lead to the speculation about a possible antianemic and cardioprotective effect of P. granatum. PMID:26881853

  4. Plasma functionalization of polycarbonaturethane to improve endothelialization--Effect of shear stress as a critical factor for biocompatibility control.

    Lukas, Karin; Thomas, Ulrich; Gessner, André; Wehner, Daniel; Schmid, Thomas; Schmid, Christof; Lehle, Karla


    Medical devices made of polycarbonaturethane (PCU) combine excellent mechanical properties and little biological degradation, but restricted hemocompatibility. Modifications of PCU might reduce platelet adhesion and promote stable endothelialization. PCU was modified using gas plasma treatment, binding of hydrogels, and coupling of cell-active molecules (modified heparin, anti-thrombin III (ATIII), argatroban, fibronectin, laminin-nonapeptide, peptides with integrin-binding arginine-glycine-aspartic acid (RGD) motif). Biocompatibility was verified with static and dynamic cell culture techniques. Blinded analysis focused on improvement in endothelial cell (EC) adhesion/proliferation, anti-thrombogenicity, reproducible manufacturing process, and shear stress tolerance of ECs. EC adhesion and antithrombogenicity were achieved with 9/35 modifications. Additionally, 6/9 stimulated EC proliferation and 3/6 modification processes were highly reproducible for endothelialization. The latter modifications comprised immobilization of ATIII (A), polyethyleneglycole-diamine-hydrogel (E) and polyethylenimine-hydrogel connected with modified heparin (IH). Under sheer stress, only the IH modification improved EC adhesion within the graft. However, ECs did not arrange in flow direction and cell anchorage was restricted. Despite large variation in surface modification chemistry and improved EC adhesion under static culture conditions, additional introduction of shear stress foiled promising preliminary data. Therefore, biocompatibility testing required not only static tests but also usage of physiological conditions such as shear stress in the case of vascular grafts. PMID:26762398

  5. Hemostatic abnormalities in liver cirrhosis

    Kendal YALÇIN


    Full Text Available In this study, 44 patients with liver cirrhosis were investigated for hemostatic parameters. Patients with spontaneous bacterial peritonitis, hepatocellular carcinoma, hepatorenal syndrome and cholestatic liver diseases were excluded. Patients were classified by Child-Pugh criterion and according to this 4 patients were in Class A, 20 in Class B and 20 in C. Regarding to these results, it was aimed to investigate the haematological disturbances in liver cirrhotic patients.In the result there was a correlation between activated partial thromboplastin time, serum iron, ferritin, transferrin, haptoglobin and Child-Pugh classification. Besides there was no correlation between prothrombin time, factor 8 and 9, protein C and S, anti-thrombin 3, fibrinogen, fibrin degradation products, serum iron binding capacity, hemoglobin, leukocyte, mean corpuscular volume and Child-Pugh classification.There were significant difference, in terms of AST, ferritin, haptoglobulin, sex and presence of ascites between groups (p0.05. In the summary, we have found correlation between hemostatic abnormalities and disease activity and clinical prognosis in patients with liver cirrhosis which is important in the management of these patients. This is also important for identification of liver transplant candidiates earlier.

  6. Heparin coating of tantalum coronary stents reduces surface thrombin generation but not factor IXa generation.

    Blezer, R; Cahalan, L; Cahalan, P T; Lindhout, T


    In the present study we used an in-vitro technique to examine initiation and propagation of blood coagulation at the surface of tantalum coronary stents exposed to flowing platelet-rich and platelet-free plasma. The time course of factor IXa production at the surface of the stent was not influenced by platelets. In spite of a significant factor IXa production, no thrombin activity was detected when the tantalum stent was exposed to platelet-free plasma; only when the stent was exposed to platelet-rich plasma was extensive thrombin production observed. These findings indicate that tantalum triggers blood coagulation, but that (adherent) platelets are essential for thrombin generation. Heparin-coated tantalum stents exposed to flowing platelet-rich plasma showed that factor IXa generation was slightly reduced compared with the bare stent. However, the heparin coating drastically delayed the onset of thrombin generation and largely reduced the steady-state production of thrombin. We found a clear relationship between the antithrombin binding capacity and the antithrombogenic potential of the heparin-coated stents. The mode of action of immobilized heparin is thought to abrogate thrombin generation by inhibiting thrombin-dependent positive feedback reactions at the surface of the coronary stent. PMID:9712292

  7. 3,4,9,10-Perylenetetracarboxylic dianhydride functionalized graphene sheet as labels for ultrasensitive electrochemiluminescent detection of thrombin.

    Gan, Xianxue; Yuan, Ruo; Chai, Yaqin; Yuan, Yali; Cao, Yaling; Liao, Yuhong; Liu, Huijing


    A novel tracer, 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA) functionalized graphene sheet (GS) composite (GS-TCDA), is employed to label the secondary anti-thrombin aptamer (TBA) to construct an ultrasensitive electrochemiluminescent sandwich-type aptasensor. The GS provided large surface area for loading abundant PTCDA and TBA with good stability and biocompatibility. Because of the excellent electroconductivity of GS and the desirable optical properties of PTCDA, the as-formed Apt II bioconjugate considerably amplified the electrochmiluminescence (ECL) signal of peroxydisulfate (S(2)O(8)(2-)) and worked as the desirable label for Apt II. On the basis of the considerably amplified ECL signal and sandwich format, an extremely wide range from 1 fM to 1 nM with an ultralow detection limit of 0.33 fM for thrombin was obtained. Additionally, the selectivity and stability of the proposed aptasensor were also excellent. Thus, this procedure has great promise for detection of thrombin present at ultra-trace levels during early stage of diseases. PMID:22541015

  8. [A sudden rise in INR due to combination of Tribulus terrestris, Avena sativa, and Panax ginseng (Clavis Panax)].

    Turfan, Murat; Tasal, Abdurrahman; Ergun, Fatih; Ergelen, Mehmet


    Warfarin sodium is an antithrombin agent used in patients with prosthetic valve and atrial fibrillation. However, there are many factors that can change the effectiveness of the drug. Today, herbal mixtures promoted through targeted print and visual media can lead to sudden activity changes in patients using warfarin. In this case report we will present two cases with a sudden rise in INR due to using combination of Tribulus terrestris, Avena sativa and Panax ginseng (Panax Clavis). Two patients who used warfarin due to a history of aortic valve replacement (case 1) and atrial fibrillation (case 2) were admitted to the hospital due very high levels of INR detected during routine follow-up. Both patients had used an herbal medicine called ''Panax'' during the last month. The patients gave no indication regarding a change in diet or the use of another agent that might interact with warfarin. In cases where active bleeding could not be determinated, we terminated the use of the drug and re-evaluated dosage of warfarin before finally discharging the patient. PMID:22864323

  9. The coagulopathy and thrombotic risk associated with L-asparaginase treatment in adults with acute lymphoblastic leukaemia.

    Truelove, E; Fielding, A K; Hunt, B J


    The dramatic improvements seen in the outcome of paediatric patients with acute lymphoblastic leukaemia (ALL) have led to increasing incorporation of L-asparaginase (L-Asp) in adult treatment protocols. However, its use is associated with a disruption in the physiological balance between haemostatic and anticoagulant pathways, with the predominant clinical manifestation being thrombosis. Although L-Asp therapy is known to be associated with an acquired deficiency of antithrombin (AT), the concurrent depletion of fibrinogen and other haemostatic proteins means that the precise mechanism of thrombosis remains to be defined. In vitro coagulation assays are often prolonged but thrombosis rather than haemorrhage is the primary concern. Management of thrombotic events in these patients is based around agents that rely on AT for their anticoagulant effect, even though it is usually depleted. There is currently only limited evidence supporting the use of AT concentrates in either primary prevention or management following an established event. Evidence-based guidelines for prevention and management strategies are lacking. PMID:23099335

  10. Glycoprotein fucosylation is increased in seminal plasma of subfertile men

    Beata Olejnik


    Full Text Available Fucose, the monosaccharide frequent in N- and O-glycans, is a part of Lewis-type antigens that are known to mediate direct sperm binding to the zona pellucida. Such interaction was found to be inhibited in vitroby fucose-containing oligo- and polysaccharides, as well as neoglycoproteins. The objective of this study was to screen seminal plasma proteins of infertile/subfertile men for the content and density of fucosylated glycoepitopes, and compare them to samples of fertile normozoospermic subjects. Seminal proteins were separated in polyacrylamide gel electrophoresis and blotted onto nitrocellulose membrane and probed with fucose-specific Aleuria aurantia lectin (AAL. Twelve electrophoretic bands were selected for quantitative densitometric analysis. It was found that the content, and especially the density of fucosylated glycans, were higher in glycoproteins present in seminal plasma of subfertile men. No profound differences in fucosylation density were found among the groups of normozoospermic, oligozoospermic, asthenozoospermic, and oligoasthenozoospermic subfertile men. According to the antibody probing, AAL-reactive bands can be attributed to male reproductive tract glycoproteins, including prostate-specific antigen, prostatic acid phosphatase, glycodelin and chorionic gonadotropin. Fibronectin, α1 -acid glycoprotein, α1 -antitrypsin, immunoglobulin G and antithrombin III may also contribute to this high fucosylation. It is suggested that the abundant fucosylated glycans in the sperm environment could interfere with the sperm surface and disturb the normal course of the fertilization cascade.

  11. Extensive cerebral venous thrombosis in a renal allograft recipient

    An increased risk of venous thromboembolism has been demonstrated following renal transplantation. Commonly reported sites have been deep vein thrombosis, pulmonary thromboembolism and vascular thrombosis involving the graft. Cerebral venous thrombosis (CVT) has not been reported in literature so far. A 36-year-old male patient, transplanted in January 2005 with normal graft functions, was admitted with history of headache, blurring of vision and vomiting. Examination revealed papilledema and no neurological deficits. Baseline investigations and analysis of cerebrospinal liquid were normal. Cerebral magnetic resonance venogram revealed extensive CVT involving superior sagittal sinus, bilateral transverse sinuses and the right sigmoid sinus. He was investigated for a thrombophilic disorder; serum homocysteine, protein C and S levels, antiphospholipid antibody and antithrombin-III levels were done despite which no conclusive diagnosis could be arrived at. To our knowledge, this is the first report of extensive CVT described in a transplant recipient. Ne definite prothrombotic or predisposing factors could be identified in our patient and the cause of CVT remains unclear. (author)

  12. Is the size of an abdominal aortic aneurysm associated with coagulopathy?

    Shindo, Shunya; Matsumoto, Harunobu; Kubota, Kenji; Kojima, Atsuo; Matsumoto, Masahiko; Satoh, Kaneo; Ozaki, Yukio


    Abdominal aortic aneurysm (AAA) volume and intraluminal thrombi were analyzed with respect to the number and function of platelets, blood cells, and coagulation factors. A group of 43 patients who underwent repair of an AAA were enrolled in this study. The maximum diameter and volume of the AAA, and the volume of intraluminal thrombi and lumen were measured by computed tomography with planimetry. The platelet count and platelet function, prothrombin time, activated partial thromboplastin time, fibrinogen, plasminogen, antithrombin 3, fibrin degradation products (FDP), D-dimer, and blood cell counts were measured. Spontaneous platelet aggregation and the FDP, and D-dimer levels were elevated; all other factors remained within the normal range. Intraluminal thrombus volume was strongly correlated with the volume and diameter of the AAA. However, no correlation was observed between the size of the AAA and coagulating factors, including the number and aggregation value of platelets. AAAs are frequently associated with a coagulating disorder. However, its size and thrombus volume are not correlated with coagulation changes. Although an intraluminal thrombus increases along with fee enlargement of the AAA, the clinical manifestation of bleeding is rarely associated with an AAA. Therefore coagulopathy in patients with an AAA is not fully explained by its morphology. PMID:15951938

  13. Antiplatelet treatment and prothrombotic diathesis following endovascular abdominal aortic aneurysm repair.

    Trellopoulos, G; Georgiadis, G S; Nikolopoulos, E S; Kapoulas, K C; Georgakarakos, E I; Lazarides, M K


    Prothrombotic diathesis expressed by elevated levels of coagulation-specific biomarkers has been reported in patients with abdominal aortic aneurysm (AAA) and after AAA endovascular repair (EVAR). This study investigates the effect of antiplatelet agents (APLs) on the prothrombotic diathesis in the post-EVAR period. Forty elective EVAR patients had thrombin-antithrombin complex, d-dimer, fibrinopeptide A, and high-sensitivity C-reactive protein measured before, at 24 hours, 1 month, and 6 months after EVAR. Patients receiving APLs postoperatively were compared with those not receiving APLs. All biomarkers were above the normal limits preoperatively and increased significantly 24 hours postoperatively followed by a drop at 1 and 6 months. No statistically significant changes were noted among patients receiving APLs in comparison with those not receiving APLs. The preoperative and postoperative prothrombotic diathesis of AAA following EVAR was confirmed in line with other reports. There was however no significant alteration of the examined biomarkers in patients receiving APLs. PMID:24101707

  14. Coagulation and fibrinolysis after open infrarenal abdominal aortic aneurysm repair in a long-term perspective.

    Holmberg, A; Bergqvist, D; Siegbahn, A


    In patients with abdominal aortic aneurysms (AAA) the coagulation and fibrinolytic systems have been found to be activated preoperatively. Does the increased activity of the coagulation and fibrinolytic systems persist after AAA surgery in a long-term perspective? Prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), tissue plasminogen activator (tPA), human plasminogen activator inhibitor type 1, and human cross-linked fibrin degradation product (D-dimer) were analysed in 18 patients after open AAA surgery (postop-AAA). The median time between surgery and blood sampling was 19 months (range, 5-37 months). Comparisons were made with both preoperative values of 23 patients with AAA (preop-AAA) as well as 20 age-matched healthy controls (AMC). F1+2, TAT, and D-dimer in preop-AAA were significantly higher compared to AMC (pD-dimer). However, TAT and D-dimer levels were still higher in postop-AAA than in AMC (paneurysmal sac but still higher than in a nonaneurysmal aorta. PMID:10574587

  15. Compensated activation of coagulation in patients with abdominal aortic aneurysm: effects of heparin treatment prior to elective surgery.

    Jelenska, Maria Magdalena; Szmidt, Jacek; Bojakowski, Krzystof; Grzela, Tomasz; Palester-Chlebowczyk, Magorzata


    Elective surgery of abdominal aortic aneurysm (AAA) sometimes leads to excessive bleeding and disseminated intravascular coagulation (DIC), even in patients with normal preoperative coagulation parameters. Coagulation screen, performed routinely before surgery is of limited value in the assessment of compensated activation of the haemostatic system. In this study, we used a number of additional tests (D-dimer, prothrombin fragment 1+2, antithrombin, and activation of fibrinolysis in the platelet poor plasma) for the diagnosis of compensated activation of the haemostatic system in AAA-patients. D-dimer and marker of thrombin generation (prothrombin fragment 1+2) positively correlated with each other (r = 0.768, P D-dimer and prothrombin fragment 1+2 decreased significantly) and resulted in the increase of platelet number and fibrinogen concentration, indicating their previous consumption. Despite differences in aneurysm diameters between the groups of 15 LMWH treated patients (mean 70.9 +/- 16 mm) and the reference group of 20 untreated AAA patients (mean 52.3 +/- 8.0 mm), intraoperative parameters (operation time, blood loss and transfusion demands) were similar. PMID:15543326

  16. Treatment of Venous Thromboembolism With New Anticoagulant Agents.

    Becattini, Cecilia; Agnelli, Giancarlo


    Venous thromboembolism (VTE) is a common disease associated with high risk for recurrences, death, and late sequelae, accounting for substantial health care costs. Anticoagulant agents are the mainstay of treatment for deep vein thrombosis and pulmonary embolism. The recent availability of oral anticoagulant agents that can be administered in fixed doses, without laboratory monitoring and dose adjustment, is a landmark change in the treatment of VTE. In Phase III trials, rivaroxaban, apixaban, edoxaban (antifactor Xa agents), and dabigatran (an antithrombin agent) were noninferior and probably safer than conventional anticoagulation therapy (low-molecular-weight heparin followed by vitamin K antagonists). These favorable results were confirmed in specific patient subgroups, such as the elderly and fragile. However, some patients, such as those with cancer or with intermediate- to high-risk pulmonary embolism, were underrepresented in the Phase III trials. Further clinical research is required before new oral anticoagulant agents can be considered standard of care for the full spectrum of patients with VTE. PMID:27102510

  17. Changes in the coagulation profile of cattle with left abomasal displacement.

    Sobiech, P; Radwińska, J; Krystkiewicz, W; Snarska, A; Stopyra, A


    The purpose of this study was to determine changes in coagulation profile parameters in cattle with left abomasal displacement (LAD). The study was performed on 20 Holstein-Friesian (H-F) cows divided into two groups: group I--10 cows with diagnosed left abomasal displacement and group II--10 clinically healthy cows. Coagulation tests, including TT (thrombin time), PT (prothrombin time) and APTT (activated partial thromboplastin time), were conducted, and fibrinogen content, D-dimer content, AT III (antithrombin III) activity and platelet (PLT) count were determined in all the animals. Prolonged TT, PT and APTT, a higher fibrinogen and D-dimer content, a drop in AT III activity and thrombocyte count were observed in the cattle with LAD. The above abnormal coagulation profiles were most predominant in three cows which died after surgical repositioning of the abomasum. The results of the study indicate that in cattle with abomasal displacement, the disseminated intravascular coagulation (DIC) syndrome was the most significant risk factor for mortality. PMID:19227127

  18. A study of some aspects of auto immunity in children with thalassemia major

    This study is subjected to estimate the study of the most common autoimmune complications in thalassaemic and the prevalence of different auto anti-bodies so as to evaluate its correlation with personal data including age, sex, growth parameters, Hepatitis C virus infection ferritin level and type of iron chelation therapy methodology : Different Abs including anti-cardiolipin, anti-tistone, ANA, Anti-erythropoietin by ELISA technique Hepatitis C virus by ELISA. Ferritin by IRMA (Immune radiometric assay), Protein C, S. Anti-thrombin III by ELISA result of the study were analyzed by appropriate statistical methods .The results showed that age of Hepatitis C virus negative cases was significantly lower than Hepatitis C virus Positive cases, ferritin levels were significantly higher in patients on defripone than those as desferrioxamine, anti-erythropoietin was detected in 93.3% of cases regarding the antithrombotic factors deficiency. Protein C was deficient in 26.67% of cases. Protein S was deficient in 13.33% only and no cases showed anti thrombin III deficiency. No special relation was found between these abnormalities and type of chelation , Conclusions: Auto-immunity may be a cofactor in the development of life threatening complication

  19. Factor V Leiden mutation does not affect coagulopathy or outcome in lethal H1N1 influenza.

    Schouten, M; van der Sluijs, K F; Roelofs, J J T H; Levi, M; Van't Veer, C; van der Poll, T


    Influenza A is a major cause of mortality. Knowledge on coagulation activation in influenza infection is limited. The factor V Leiden (FVL) mutation is possibly subject to positive selection pressure. It is unknown whether this mutation impacts on the outcome of severe influenza. In the present study, the effect of lethal influenza on pulmonary and systemic coagulation activation and whether or not FVL mutation alters coagulation activation in and the course of lethal influenza, was determined. Wild-type mice, and mice heterozygous or homozygous for FVL were infected intranasally with a lethal dose of H1N1 (haemagglutinin 1 and neuraminidase 1) influenza A. Mice were sacrificed after 48 or 96 h for determination of coagulation activation, histopathology, pulmonary inflammatory parameters and viral load, or were observed in a survival study. Extensive local and systemic coagulation activation during lethal influenza was demonstrated by increased lung and plasma levels of thrombin-antithrombin complexes and fibrin degradation products, and by pulmonary fibrin deposition. FVL mutation did not influence the procoagulant response, lung histopathology or survival. FVL mice demonstrated elevated viral loads 48 h after infection. In conclusion, coagulation is activated locally and systemically during lethal murine influenza A infection. The FVL mutation does not influence coagulation activation, lung inflammation or survival in lethal influenza A. PMID:20413539

  20. [Biological and clinical safety of nomegestrol acetate administered alone then associated in inverse sequence with transdermal 17 beta estradiol, in women at risk of dyslipoproteinemia type IIa].

    Zartarian, M; Chevallier, T; Micheletti, M C; Leber, C; Jamin, C


    In this study including 26 patients with dyslipoproteinemia classified IIa, we evaluated biochemical and clinical safety of Nomegestrol acetate (Lutenyl) used for its antigonadotrophin property. It was administered alone, during 3 cycles at the dose of 5 mg/d for 21 days by cycle and then it was associated (at the same sequence and dose), without any wash out, for the next 6 cycles, with a 17 beta estradiol patch (Estraderm TTS 50), 50 micrograms/d from the 11th to the 21st day of each cycle. Nomegestrol acetate, alone, had no significant effect on glycemia, antithrombin III, triglycerides, total cholesterol, apoprotein A1, and LpA1 values compared to those at baseline but apoprotein B and Lp (a) values tended to decrease slightly. Serum progesterone levels were collapsed, and FSH values were low. Weight and blood pressure remained constant. Adding 17 beta estradiol enabled to significantly decrease and normalize the apoprotein B values after the first 3 cycles compared to the baseline values, then these values remained constant during the next 3 cycles. There was no effect on the other parameters (except for a significant increase in plasmatic estradiol values) on the antigonadotrophin property of Nomegestrol acetate, nor on weight and blood pressure which remained constant. Moreover, we observed an important decrease in the rate of amenorrheic cycles compared to those with Nomegestrol acetate alone. PMID:9949893

  1. Fondaparinux sodium.

    Keam, Susan J; Goa, Karen L


    black triangle Fondaparinux sodium, a selective factor Xa inhibitor, is the first in a new class of antithrombotics. It binds selectively with high affinity to antithrombin III and specifically catalyses the inactivation of factor Xa. The elimination half-life of fondaparinux sodium permits once daily treatment. black triangle A randomised, double-blind, parallel-group, dose-ranging, multicentre phase IIb study in 933 eligible patients established that a subcutaneous dose of between 1.5 and 3mg of fondaparinux sodium has the optimum efficacy and safety profile for prophylaxis of venous thromboembolism in patients undergoing major orthopaedic surgery. black triangle Fondaparinux sodium, given to more than 3600 patients undergoing major orthopaedic surgery who participated in prospective, randomised, double-blind, multicentre phase III clinical trials, significantly reduced the incidence of venous thromboembolism, with an overall risk reduction of 55.2% compared with enoxaparin. black triangle Fondaparinux sodium was well tolerated by patients undergoing major orthopaedic surgery, and at the recommended clinical dose of 2.5mg has a similar tolerability profile, including bleeding events, to standard enoxaparin regimens. Fondaparinux sodium has not been reported to cause antibody-induced thrombocytopenia. PMID:12109927

  2. Structural features and inactivation of coagulation proteases of a sulfated polysaccharidic fraction from Caulerpa cupressoides var. lycopodium (Caulerpaceae, Chlorophyta - doi: 10.4025/actascitechnol.v35i4.16709

    José Ariévilo Gurgel Rodrigues


    Full Text Available Studies on biopolymers from macroalgae suggested sulfated polysaccharides (SPs as research agents to investigate events related to haemostasis. Caulerpa cupressoides var. lycopodium is a marine green alga containing three SPs fractions (SP1, SP2 and SP3. SP2 had anticoagulant (in vitro and anti- and prothrombotic (in vivo actions; however, its effect on the coagulation system is not fully understood. This study aimed to determine the infrared (IR spectroscopy, chemical composition (CC, elemental microanalysis (EM, molecular weight (MW and the effect on coagulation proteases of SP2. The presence of sulfate ester, galactose-6-sulfate, uronic acid and glycoside linkages for IR spectrum; contents of sulfate (28%, total sugars (40% and uronic acids (7.18% for CC; and content of carbon (21.98%, sulfate (4.27%, nitrogen (1.3% and hydrogen (4.86% for EM were obtained. The average molecular weights of four different SPs (SP-1, SP-2, SP-3 and SP-4 subfractions from the SP2 ranged from ~ 8 to >100 kDa. SP2 was tested on coagulation proteases (thrombin and factor Xa in the presence of antithrombin (AT and heparin cofactor II (HCII using human plasma, being both thrombin and factor Xa target proteases inhibited, but requiring a concentration of about 2.5-fold higher of HCII than the thrombin inactivation by AT.   

  3. Leech therapeutic applications

    A M Abdualkader


    Full Text Available Hematophagous animals including leeches have been known to possess biologically active compounds in their secretions, especially in their saliva. The blood-sucking annelids, leeches have been used for therapeutic purposes since the beginning of civilization. Ancient Egyptian, Indian, Greek and Arab physicians used leeches for a wide range of diseases starting from the conventional use for bleeding to systemic ailments, such as skin diseases, nervous system abnormalities, urinary and reproductive system problems, inflammation, and dental problems. Recently, extensive researches on leech saliva unveiled the presence of a variety of bioactive peptides and proteins involving antithrombin (hirudin, bufrudin, antiplatelet (calin, saratin, factor Xa inhibitors (lefaxin, antibacterial (theromacin, theromyzin and others. Consequently, leech has made a comeback as a new remedy for many chronic and life-threatening abnormalities, such as cardiovascular problems, cancer, metastasis, and infectious diseases. In the 20 th century, leech therapy has established itself in plastic and microsurgery as a protective tool against venous congestion and served to salvage the replanted digits and flaps. Many clinics for plastic surgery all over the world started to use leeches for cosmetic purposes. Despite the efficacious properties of leech therapy, the safety, and complications of leeching are still controversial.

  4. Hematologic and hemostatic changes induced by different columns during LDL apheresis.

    Hovland, Anders; Hardersen, Randolf; Nielsen, Erik Waage; Mollnes, Tom Eirik; Lappegård, Knut Tore


    Low density lipoprotein (LDL) apheresis is a long-term treatment and its impact on risk factors other than lipoproteins could be of importance. Three patients with familial hypercholesterolemia participated in six consecutive treatments with three different LDL apheresis columns in random order: DL-75, LA-15, and EC-50W. We compared treatment effects on hemoglobin, leukocytes, platelets, fibrinogen, thrombin-antithrombin complexes (TAT), plasminogen activator inhibitor-1 (PAI-1), and homocysteine. Hemoglobin, leukocytes and platelets decreased significantly with DL-75 (P < 0.05). Hemoglobin and leukocytes increased significantly with LA-15 and EC-50W (P < 0.05). Platelets were unchanged. The DL-75 column was statistically different from LA-15 and EC-50W regarding these parameters. With the columns DL-75, LA-15, and EC-50W fibrinogen decreased significantly by 28%, 32%, and 42%, PAI-1 decreased significantly by 72%, 58%, and 30% while TAT increased significantly by 138%, 3%, and 251%, respectively (P < 0.05 for all). When comparing the columns there were significant differences between all of them regarding fibrinogen, no differences regarding TAT and a difference between DL-75 and EC-50W regarding PAI-1. With the columns DL-75, LA-15 and EC-50W homocysteine decreased 22%, 9%, and 13%, respectively, but there were no inter column differences. In conclusion, the three LDL apheresis columns affected important hematological and hemostatic risk factors differently. PMID:20806414

  5. Coagulation disorders in the patients with deep vein thrombosis of lower extremity

    Milić Dragan J.


    Full Text Available PURPOSE Venous thromboembolism is a relevant social and health care problem for its high incidence, pulmonary embolism-related mortality and long-term sequelae which may be disabling (post-thrombotic syndrome and ulceration. PROCEDURES The aim of our work was to establish the presence of coagulation disorders (hypercoagulable states in the patients with deep vein thrombosis (DVT of the leg. Prospectively we have analyzed a group of 30 patients with echosono-graphicaly verified DVT of the leg who were admitted to the department of vascular surgery from August 1st 2000 to July 31st 2001.The following parameters were monitored: prothrombin time (PT partial thromboplastin time (PTT, fibrinogen (Fib, alpha 2 antiplasmin (A-2 AP, D-dimer (DD, antithrombin III (AT III and factor VII. FINDINGS Activation of the coagulation process was registered. The values of monitored coagulation parameters are shown in table 1. Plasma levels of monitored parameters in the patients with DVT of the leg were significantly higher than in the control subjects. CONCLUSION In patients with a DVT a hypercoagulable state is common finding. Some parameters of coagulation activity such as D-dimer might be of great interest in the diagnostic strategy of DVT.

  6. What neurosurgeons need to know about dabigatran etexilate (pradax®/pradaxa®/prazaxa®)

    Dwyer, Christopher Michael; Damodaran, Omprakash; Heckelmann, Michael; Sheridan, Mark Michael


    Dabigatran etexaliate is a novel oral anticoagulant that directly inhibits thrombin. It offers a number of substantial medical benefits over other oral and parenteral anticoagulants but its advent raises important neurosurgical considerations. Dabigatran has important potential benefits. Unlike warfarin, it does not require routine blood tests to monitor its anticoagulative effect and there is no need for dose titration. Drug interactions are greatly simplified when compared to warfarin as dabigatran is not metabolized by cytochrome p450 isoenzymes. As a result, dabigatran has been approved in many jurisdictions for DVT prophylaxis after orthopaedic surgery and also for the prevention of embolic events associated with non-valvular atrial fibrillation. There are, however, important neurosurgical challenges associated with regular dabigatran use. Unlike current anti-coagulants, there is no specific reversal agent for dabigatran. Known reversal options include activated charcoal (within one to two hours of intake) and renal dialysis. Protamine sulfate and vitamin K are unlikely to affect the activity of dabigatran. Platelet concentrates will not inactivate dabigatran's anti-thrombin properties. Assessing the degree of anticoagulation is difficult as conventional markers of serum coagulability are typically normal in patients taking dabigatran. The potential neurosurgical challenges of dabigatran were cast in sharp relief by a recent case report from the United States that is considered in this note. In the absence of a clear reversal pathway, we propose a treatment algorithm for chronic dabigatran use based on the replacement of any deficient factors and rapid access to renal dialysis. PMID:25972932

  7. What neurosurgeons need to know about dabigatran etexilate (pradax(®)/pradaxa(®)/prazaxa(®)).

    Dwyer, Christopher Michael; Damodaran, Omprakash; Heckelmann, Michael; Sheridan, Mark Michael


    Dabigatran etexaliate is a novel oral anticoagulant that directly inhibits thrombin. It offers a number of substantial medical benefits over other oral and parenteral anticoagulants but its advent raises important neurosurgical considerations. Dabigatran has important potential benefits. Unlike warfarin, it does not require routine blood tests to monitor its anticoagulative effect and there is no need for dose titration. Drug interactions are greatly simplified when compared to warfarin as dabigatran is not metabolized by cytochrome p450 isoenzymes. As a result, dabigatran has been approved in many jurisdictions for DVT prophylaxis after orthopaedic surgery and also for the prevention of embolic events associated with non-valvular atrial fibrillation. There are, however, important neurosurgical challenges associated with regular dabigatran use. Unlike current anti-coagulants, there is no specific reversal agent for dabigatran. Known reversal options include activated charcoal (within one to two hours of intake) and renal dialysis. Protamine sulfate and vitamin K are unlikely to affect the activity of dabigatran. Platelet concentrates will not inactivate dabigatran's anti-thrombin properties. Assessing the degree of anticoagulation is difficult as conventional markers of serum coagulability are typically normal in patients taking dabigatran. The potential neurosurgical challenges of dabigatran were cast in sharp relief by a recent case report from the United States that is considered in this note. In the absence of a clear reversal pathway, we propose a treatment algorithm for chronic dabigatran use based on the replacement of any deficient factors and rapid access to renal dialysis. PMID:25972932

  8. The relationship between thrombophilic mutations and preeclampsia: a prospective case-control study

    Preeclampsia and its association with thrombophillia remain controversial, due to inconsistent results in different studies, which different ethnic groups, selection criteria, and patient numbers. The aim of this study was to determine the relationship between thrombophillia and preeclamptic patients in our region. In a prospective case-control study, we compared 100 consecutive women with preeclampsia and eclampsia (group 1) with 100 normal pregnant women (group 2). All women were tested two months after delivery for mutations of factor V Leiden, methylenetetrahydrofolate reductase (MTHFR), and prothrombin gene mutation mutataion as well as for deficiencies of protein C, protein S, and antithrmbin III. A thrombophilic mutation was found in 42 (42%) and 28(28%) women in group I and group II, respectively (P+0.27, OR 1.5, 95% CI 1.0-2.2). The incidence of Factor V Leiden mutation (heterozygous), prothrombin mutation (heterozygous), prothrombin mutation (homozygous), MTHFR mutation (homozygous) was not statistically significant in group 1 compared with group 2 (P>0.05). Also, deficiencies of protein S, protein c and antithrombin III were not statistically significant in group I com pared with group II (P>0.05). There was no difference in thrombophilic mutations between preeclamptic patients and normal pregnant women in our region. Therefore, we suggest that preeclamptic patients should not be tested for thrombophilia. (author)

  9. Lack of Association between Recurrent Pregnancy Loss and Inherited Thrombophilia in a Group of Colombian Patients

    Henry Cardona


    Full Text Available Studies have shown an association between recurrent pregnancy loss and inherited thrombophilia in Caucasian populations, but there is insufficient knowledge concerning triethnic populations such as the Colombian. The aim of this study was to evaluate whether inherited thrombophilia is associated with recurrent pregnancy loss. Methods. We conducted a case-control study of 93 patients with recurrent pregnancy loss (cases and 206 healthy multiparous women (controls in a Colombian subpopulation. Three single nucleotide polymorphisms (SNPs markers of the inherited thrombophilias factor V Leiden, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T were genotyped by PCR-RFLP. Activated protein C resistance and plasma levels of antithrombin, protein C, and protein S were also measured. Results. The frequency of thrombophilia-associated SNPs, activated protein C resistance, and anticoagulant protein deficiencies, was low overall, except for the methylenetetrahydrofolate reductase C677T SNP. The differences between patients and controls had no statistical significance. Conclusion. Our study confirms the low prevalence of inherited thrombophilias in non-Caucasian populations and it is unlikely that the tested thrombophilias play a role in the pathogenesis of recurrent pregnancy loss in this Colombian population.

  10. Predilution versus postdilution during continuous venovenous hemofiltration: a comparison of circuit thrombogenesis.

    de Pont, Anne-Cornélie J M; Bouman, Catherine S C; Bakhtiari, Kamran; Schaap, Marianne C L; Nieuwland, Rienk; Sturk, Augueste; Hutten, Barbara A; de Jonge, Evert; Vroom, Margreeth B; Meijers, Joost C M; Büller, Harry R


    During continuous venovenous hemofiltration, predilution can prolong circuit survival time, but the underlying mechanism has not been elucidated. The aim of the present study was to compare predilution with postdilution, with respect to circuit thrombogenesis. Eight critically ill patients were treated with both predilutional and postdilutional continuous venovenous hemofiltration in a crossover fashion. A filtration flow of 60 ml/min was used in both modes. We chose blood flows of 140 and 200 ml/min during predilution and postdilution, respectively, to keep the total flow through the hemofilter constant. Extracorporeal circuit pressures were measured hourly, and samples of blood and ultrafiltrate were collected at five different time points. Thrombin-antithrombin complexes and prothrombin fragments F1 + 2 were measured by ELISA, and platelet activation was assessed by flow cytometry. No signs of thrombin generation or platelet activation were found during either mode. During postdilution, baseline platelet count and maximal prefilter pressure had a linear relation, whereas both parameters were inversely related with circuit survival time. In summary, predilution and postdilution did not differ with respect to extracorporeal circuit thrombogenesis. During postdilution, baseline platelet count and maximal prefilter pressure were inversely related with circuit survival time. PMID:16883122

  11. [Complex assessment of vasomotor function of vascular endothelium in patients with hypertension].

    Gel'tser, B I; Savchenko, S V; Kotel'nikov, V N; Plotnikova, I V


    Vasomotor function of vascular endothelium was studied in 62 patients with grade 1-2 hypertension with moderate and high added risk. Methods included study of brachial and middle cerebral artery endothelium dependent and independent vasodilation/vasoconstriction, measurement of plasma levels of nitric oxide metabolites (NO(n)-), endothelin-1, and antithrombin, as well as registration of their changes during vasomotor tests with calculation of integral indexes. Most patients with hypertension differed from controls by preponderance of vasoconstrictor over vasodilator reactions both in peripheral and cerebral vascular bed. At the same time patients with hypertension had pronounced dissociation between vasomotor responses of cerebral and peripheral vessels compared with subjects with normal blood pressure (pconcentration of endothelin-1 patients with hypertension were characterized by hyperreactivity of nitricoxidergic system, augmented lability of endothelin producing system, and impaired athrombogenecity of vascular endothelium. Complex assessment of vasomotor function of vascular endothelium by sequential vasoactive tests characterizes functional and metabolic activity of cerebral and peripheral vessels and can be used for improvement of risk stratification and monitoring of efficacy of treatment of patients with hypertension. PMID:15111971

  12. Robustness of nanofiltration for increasing the viral safety margin of biological products.

    Caballero, Santiago; Diez, José M; Belda, Francisco J; Otegui, Magdalena; Herring, Steven; Roth, Nathan J; Lee, Douglas; Gajardo, Rodrigo; Jorquera, Juan I


    In this study, the virus-removal capacity of nanofiltration was assessed using validated laboratory scale models on a wide range of viruses (pseudorabies virus; human immunodeficiency virus; bovine viral diarrhea virus; West Nile virus; hepatitis A virus; murine encephalomyocarditis virus; and porcine parvovirus) with sizes from 18 nm to 200 nm and applying the different process conditions existing in a number of Grifols' plasma-derived manufacturing processes (thrombin, α1-proteinase inhibitor, Factor IX, antithrombin, plasmin, intravenous immunoglobulin, and fibrinogen). Spiking experiments (n = 133) were performed in process intermediate products, and removal was subsequently determined by infectivity titration. Reduction Factor (RF) was calculated by comparing the virus load before and after nanofiltration under each product purification condition. In all experiments, the RFs were close to or greater than 4 log10 (>99.99% of virus elimination). RF values were not significantly affected by the process conditions within the limits assayed (pH, ionic strength, temperature, filtration ratio, and protein concentration). The virus-removal capacity of nanofiltration correlated only with the size of the removed agent. In conclusion, nanofiltration, as used in the manufacturing of several Grifols' products, is consistent, robust, and not significantly affected by process conditions. PMID:24485384

  13. NCO-sP(EO-stat-PO Coatings on Gold Sensors—a QCM Study of Hemocompatibility

    Frank K. Gehring


    Full Text Available The reliability of implantable blood sensors is often hampered by unspecific adsorption of plasma proteins and blood cells. This not only leads to a loss of sensor signal over time, but can also result in undesired host vs. graft reactions. Within this study we evaluated the hemocompatibility of isocyanate conjugated star shaped polytheylene oxide—polypropylene oxide co-polymers NCO-sP(EO-stat-PO when applied to gold surfaces as an auspicious coating material for gold sputtered blood contacting sensors. Quartz crystal microbalance (QCM sensors were coated with ultrathin NCO-sP(EO-stat-PO films and compared with uncoated gold sensors. Protein resistance was assessed by QCM measurements with fibrinogen solution and platelet poor plasma (PPP, followed by quantification of fibrinogen adsorption. Hemocompatibility was tested by incubation with human platelet rich plasma (PRP. Thrombin antithrombin-III complex (TAT, β-thromboglobulin (β-TG and platelet factor 4 (PF4 were used as coagulation activation markers. Furthermore, scanning electron microscopy (SEM was used to visualize platelet adhesion to the sensor surfaces. Compared to uncoated gold sensors, NCO-sP(EO-stat-PO coated sensors revealed significant better resistance against protein adsorption, lower TAT generation and a lower amount of adherent platelets. Moreover, coating with ultrathin NCO-sP(EO-stat-PO films creates a cell resistant hemocompatible surface on gold that increases the chance of prolonged sensor functionality and can easily be modified with specific receptor molecules.

  14. Surface treatment of biomaterials by gamma and swift heavy ions grafting

    To date, the successful substitution of small diameter arteries by vascular grafts has not been achieved in humans. In order to reduce their thrombogenicity which remains the major obstacle, polymeric materials endowed with a specific affinity for Antithrombin III (ATIII) and thus able to catalyse the inhibition of thrombin by ATIII, as heparin does, were devised. Sulfonate and sulfonamide groups were introduced on phenyl rings belonging to styrene residues which were radiation grafted (swift heavy ions and gamma radiation) onto poly(vinylene difluoride) (PVDF) and poly(hexafluoropropylene vinylidene fluoride) (P(VDF-HFP)). Contrary to gamma radiation grafting, with swift heavy ions grafting, only small regions are modified; thus, the properties of the initial substrate are preserved. A characterization of surface topography was carried out by Scanning Electron Microscopy and Atomic Force Microscopy. The efficacy of the surface modification was monitored with respect to the total amount of bound 'heparin-like' molecules measured by a toluidine blue assay, and the anticoagulant potential estimated by the thrombin time test

  15. Effects of carboxymethyl chitosan on the blood system of rats

    Fu, Dawei [College of Marine Life Sciences, Ocean University of China, Qingdao 266003 (China); Han, Baoqin, E-mail: [College of Marine Life Sciences, Ocean University of China, Qingdao 266003 (China); Dong, Wen; Yang, Zhao; Lv, You; Liu, Wanshun [College of Marine Life Sciences, Ocean University of China, Qingdao 266003 (China)


    Highlights: {yields} We report, for the first time, the safety of carboxymethyl chitosan in blood system. {yields} CM-Chitosan has no significant effects on coagulation function of rats. {yields} CM-Chitosan has no significant effects on anticoagulation performance of rats. {yields} CM-Chitosan has no significant effects on fibrinolytic function of rats. {yields} CM-Chitosan has no significant effects on hemorheology of rats. -- Abstract: Carboxymethyl chitosan (CM-chitosan), a derivative of chitosan, was extensively studied in the biomedical materials field for its beneficial biological properties of hemostasis and stimulation of healing. However, studies examining the safety of CM-chitosan in the blood system are lacking. In this study CM-chitosan was implanted into the abdominal cavity of rats to determine blood indexes at different times and to evaluate the effects of CM-chitosan on the blood system of rats. Coagulation function was reflected by thrombin time (TT), prothrombin time (PT), activated partial thromboplatin time (APTT), fibrinogen (FIB) and platelet factor 4 (PF4) indexes; anti-coagulation performance was assessed by the index of antithrombinIII (ATIII); fibrinolytic function was reflected by plasminogen (PLG) and fibrin degradation product (FDP) indexes; and blood viscosity (BV) and plasma viscosity (PV) indexes reflected hemorheology. Results showed that CM-chitosan has no significant effects on the blood system of rats, and provides experimental basis for CM-chitosan to be applied in the field of biomedical materials.

  16. Characterization of a novel autosomal dominant bleeding disorder in a large kindred from east Texas.

    Kuang, S Q; Hasham, S; Phillips, M D; Wolf, D; Wan, Y; Thiagarajan, P; Milewicz, D M


    A large east Texas family with autosomal dominant inheritance of a novel bleeding disorder has been identified. The disorder is characterized clinically by easy bruising, life-threatening bleeding with trauma or surgery, and menorrhagia in affected women. Laboratory studies demonstrated prolongation of the prothrombin time and activated partial thromboplastin time in affected individuals. Paradoxically, assays of known coagulation factors are all within normal limits. To determine the molecular basis of this disease, a candidate gene linkage analysis in this kindred was done. Initially it was hypothesized that the cause of the disease in this family could be an antithrombin III (AT3) mutation that resulted in a constitutively active AT3 in the absence of heparin binding. Linkage studies using DNA from the family and an intragenic polymorphic marker within the AT3 gene showed that the disease mapped to this locus. The coding region and intron/exon junctions of AT3 were sequenced using the proband's DNA, but this analysis failed to identify a mutation. Additional family members were recruited for the study, and 16 polymorphic markers around the AT3 gene were analyzed. Using 2 recombinants, the critical interval for the defective gene was narrowed to approximately 1.5 Mb, centromeric to AT3. The factor V (FV) gene was mapped into the disease interval and sequenced; there were no mutations found. Elucidation of the genetic defect causing the bleeding disorder in this family may reveal a novel protein involved in the coagulation cascade. PMID:11238089

  17. [Aneurysm of the abdominal aorta and preoperative disseminated intravascular coagulation].

    Chauvet, V; Bussac, J J; Jullian, H; Juhan-Vague, I; Branchereau, A


    A case of abdominal aortic aneurysm associated with preoperative signs of disseminated intravascular coagulation is reported. The 69-year-old female patient presented with spontaneously appearing petechiae and bruising. She had 0.95 g.l-1 fibrinogen, 105 G.l-1 platelets, and 100 fibrin and fibrinogen degradation products. Investigations revealed an 80 mm diameter aneurysm of the abdominal aorta, extending from the coeliac trunk to the iliac arteries. Heparin 7,000 resulted in a biological improvement for a week only. At that time, levels of coagulation factors were: 92% factor II, 88% factor V, 100% factors VII and X, 100% antithrombin III. Surgical cure of the aneurysm was nevertheless carried out. Twenty standard units of platelets, 8 g fibrinogen, four units of fresh frozen plasma, five homologous and two autologous red cell units were transfused during the procedure. No coagulation factors were necessary during the postoperative course, which was uneventful. The management of coagulation factor infusions, before or after aortic cross-clamping, is discussed. PMID:2058833

  18. Structure and anticoagulant properties of sulfated glycosaminoglycans from primitive Chordates



    Full Text Available Dermatan sulfates and heparin, similar to the mammalian glycosaminoglycans, but with differences in the degree and position of sulfation were previously isolated from the body of the ascidian Styela plicata and Ascidia nigra. These differences produce profound effects on their anticoagulant properties. S. plicata dermatan sulfate composed by 2-O-sulfatedalpha-L-iduronic acid and 4-O-sulfated N-acetyl-beta-D-galactosamine residues is a potent anticoagulant due to a high heparin cofactor II activity. Surprisingly, it has a lower potency to prevent thrombus formation on an experimental model and a lower bleeding effect in rats than the mammalian dermatan sulfate. In contrast, A. nigra dermatan sulfate, also enriched in 2-O-sulfated alpha-L-iduronic acid, but in this case sulfated at O-6 of the N-acetyl-beta-D-galactosamine units, has no in vitro or in vivo anticoagulant activity, does not prevent thrombus formation but shows a bleeding effect similar to the mammalian glycosaminoglycan. Ascidian heparin, composed by 2-O-sulfated alpha-L-iduronic acid, N- and 6-O-sulfated glucosamine (75% and alpha-L-iduronic acid, N- and 6-O-sulfated glucosamine (25% disaccharide units has an anticoagulant activity 10 times lower than the mammalian heparin, is about 20 times less potent in the inhibition of thrombin by antithrombin, but has the same heparin cofactor II activity as mammalian heparin.

  19. Solute removal capacity of high cut-off membrane plasma separators.

    Ohkubo, Atsushi; Kurashima, Naoki; Nakamura, Ayako; Miyamoto, Satoko; Iimori, Soichiro; Rai, Tatemitsu


    In vitro blood filtration was performed by a closed circuit using high cut-off membrane plasma separators, EVACURE EC-2A10 (EC-2A) and EVACURE EC-4A10 (EC-4A). Samples were obtained from sampling sites before the plasma separator, after each plasma separator, and from the ultrafiltrate of each separator. The sieving coefficient (S.C.) of total protein (TP), albumin (Alb), IgG, interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), fibrinogen (Fib), antithrombin III (AT-III), and coagulation factor XIII (FXIII) were calculated. The S.C. of each solute using EC-2A and EC-A4 were as follows; TP: 0.25 and 0.56, Alb: 0.32 and 0.73, IgG: 0.16 and 0.50, IL-6:0.73 and 0.95, IL-8:0.85 and 0.82, TNF-α: 1.07 and 0.99, Fib: 0 and 0, FXIII: 0.07 and 0.17, respectively. When compared with the conventional type of membrane plasma separators, EVACURE could efficiently remove cytokines while retaining coagulation factors such as fibrinogen. Moreover, EC-2A prevented protein loss, whereas EC-4A could remove approximately 50% of IgG. PMID:24107276

  20. Clinical Presentation, Management, and Outcomes of Deep Vein Thrombosis Based on Doppler Ultrasonography Examination.

    Al-Thani, Hassan; El-Menyar, Ayman; Asim, Mohammad; Kiliyanni, Abdul Salim


    We studied the frequency, clinical presentation, and outcomes of deep vein thrombosis (DVT). Serial Doppler ultrasonography was performed between 2008 and 2013 for 6420 patients with suspected DVT. Diagnosis was confirmed in 662 (10.3%) participants (mean age: 50 ± 17 years; 51% females). Obesity, diabetes mellitus, and malignancy were reported in 47%, 28%, and 16%, respectively. Abnormal protein C, protein S, factor V Leiden, or antithrombin III were found in 9%, 7%, 3.8%, and 4%, respectively. Left, right, and both legs were involved in 55%, 37%, and 8%, respectively. Common femoral, popliteal, and posterior tibial veins were affected in 48.5%, 72%, and 71%, respectively. Postthrombotic syndrome, pulmonary embolism, and death were reported in 50%, 12.2%, and 15% of cases, respectively. Kaplan-Meier survival curves and Cox regression analysis showed that gender had no impact on mortality during follow-up; however, age (>50 years) was associated with greater risk of death (hazard ratio: 6.54; 95% confidence interval: 3.2-13.3). These findings will improve our understanding of the various risk factors and help develop institutional guidelines for the management of patients with DVT. PMID:26345414

  1. Effects of Erwinia-asparaginase on the coagulation system.

    Carlsson, H; Stockelberg, D; Tengborn, L; Braide, I; Carneskog, J; Kutti, J


    L-Asparaginase treatment during induction therapy in acute lymphoblastic leukaemia (ALL) is known to be frequently complicated by thromboembolic events. It was recently suggested that L-asparaginase derived from Erwinia chrysanthemi alters the coagulation system less severely than does Escherichia coli asparaginase. In a series of 11 adult patients with ALL, we investigated some parameters of the coagulation system during treatment with Erwinia asparaginase. The doses employed were rather high; all patients below the age of 60 years received 15,000 U/m2 daily over 14 days. In accordance with what is known from treatment with E. coli asparaginase, we observed significant lowering of antithrombin as well as of fibrinogen. However, as to fibrinogen indeed a significant decrease had occurred prior to the institution of Erwinia asparaginase treatment. The most striking observation in the present study was that the levels of prothrombin complex, reflecting the function of K-vitamin dependent coagulation factors II, VII and X, remained within normal ranges during treatment. This indicates that these coagulation factors were not affected by Erwinia asparaginase, an observation at variance with several reports where E. coli asparaginase was investigated. This latter observation was the only finding which could lend support to the view that Erwinia asparaginase affects the coagulation system less than E. coli asparaginase. Finally, one of our patients developed a sinus thrombosis, a severe thrombotic complication. PMID:7493674

  2. Blood peptidome-degradome profile of breast cancer.

    Yufeng Shen

    Full Text Available BACKGROUND: Cancer invasion and metastasis are closely associated with activities within the degradome; however, little is known about whether these activities can be detected in the blood of cancer patients. METHODOLOGY AND PRINCIPAL FINDINGS: The peptidome-degradome profiles of pooled blood plasma sampled from 15 breast cancer patients (BCP and age, race, and menopausal status matched control healthy persons (HP were globally characterized using advanced comprehensive separations combined with tandem Fourier transform mass spectrometry and new data analysis approaches that facilitated top-down peptidomic analysis. The BCP pool displayed 71 degradome protein substrates that encompassed 839 distinct peptidome peptides. In contrast, the HP 50 degradome substrates found encompassed 425 peptides. We find that the ratios of the peptidome peptide relative abundances can vary as much as >4000 fold between BCP and HP. The experimental results also show differential degradation of substrates in the BCP sample in their functional domains, including the proteolytic and inhibitory sites of the plasmin-antiplasmin and thrombin-antithrombin systems, the main chains of the extracellular matrix protection proteins, the excessive degradation of innate immune system key convertases and membrane attack complex components, as well as several other cancer suppressor proteins. CONCLUSIONS: Degradomics-peptidomics profiling of blood plasma is highly sensitive to changes not evidenced by conventional bottom-up proteomics and potentially provides unique signatures of possible diagnostic utility.

  3. Macrophage Matrix Metalloproteinase-12 Dampens Inflammation and Neutrophil Influx in Arthritis

    Caroline L. Bellac


    Full Text Available Resolution of inflammation reduces pathological tissue destruction and restores tissue homeostasis. Here, we used a proteomic protease substrate discovery approach, terminal amine isotopic labeling of substrates (TAILS, to analyze the role of the macrophage-specific matrix metalloproteinase-12 (MMP12 in inflammation. In murine peritonitis, MMP12 inactivates antithrombin and activates prothrombin, prolonging the activated partial thromboplastin time. Furthermore, MMP12 inactivates complement C3 to reduce complement activation and inactivates the chemoattractant anaphylatoxins C3a and C5a, whereas iC3b and C3b opsonin cleavage increases phagocytosis. Loss of these anti-inflammatory activities in collagen-induced arthritis in Mmp12−/− mice leads to unresolved synovitis and extensive articular inflammation. Deep articular cartilage loss is associated with massive neutrophil infiltration and abnormal DNA neutrophil extracellular traps (NETs. The NETs are rich in fibrin and extracellular actin, which TAILS identified as MMP12 substrates. Thus, macrophage MMP12 in arthritis has multiple protective roles in countering neutrophil infiltration, clearing NETs, and dampening inflammatory pathways to prepare for the resolution of inflammation.

  4. Preeclampsia – will Orphan Drug Status facilitate innovative biological therapies?

    Sinuhe eHahn


    Full Text Available It is generally accepted that development of novel therapies to treat pregnancy-relates disorders, such as preeclampsia, is hampered to the paucity of research funding. Hence, it is with great interest to become aware of at least three novel therapeutic approaches for the treatment of this disorder, exploiting either the anticoagulant activity of antithrombin, the free radical scavenging activity of alpha-1-microglobulin, or the regenerative capacity of placenta-derived mesenchymal stem cells. As these projects are being carried out by small biotech enterprises, the question arises of how they are able to fund such undertakings. A novel strategy adopted by two of these companies is that they successfully petitioned US and EU agencies in order that preeclampsia be accepted in the register of rare or orphan diseases. This provides a number of benefits including market exclusivity, assistance with clinical trials and dedicated funding schemes. Other strategies to supplement meager research funds, especially to test novel approaches, could be crowdfunding, a venture which relies on intimate interaction with advocacy groups. In other words, preeclampsia meets Facebook. Perhaps similar strategies can be adopted to examine novel therapies targeting either the imbalance in angiogenic growth factors, complement activation, reduced levels of placenta protein 13 or excessive neutrophil activation evident in preeclampsia.

  5. Preeclampsia – Will Orphan Drug Status Facilitate Innovative Biological Therapies?

    Hahn, Sinuhe


    It is generally accepted that the development of novel therapies to treat pregnancy-related disorders, such as preeclampsia, is hampered by the paucity of research funding. Hence, it is with great interest to become aware of at least three novel therapeutic approaches for the treatment of this disorder: exploiting either the anticoagulant activity of antithrombin, the free radical scavenging activity of alpha-1-microglobulin, or the regenerative capacity of placenta-derived mesenchymal stem cells. As these projects are being carried out by small biotech enterprises, the question arises of how they are able to fund such undertakings. A novel strategy adopted by two of these companies is that they successfully petitioned US and EU agencies in order that preeclampsia is accepted in the register of rare or orphan diseases. This provides a number of benefits including market exclusivity, assistance with clinical trials, and dedicated funding schemes. Other strategies to supplement meager research funds, especially to test novel approaches, could be crowdfunding, a venture that relies on intimate interaction with advocacy groups. In other words, preeclampsia meets Facebook. Perhaps similar strategies can be adopted to examine novel therapies targeting either the imbalance in pro- or anti-angiogenic growth factors, complement activation, reduced levels of placenta protein 13, or excessive neutrophil activation evident in preeclampsia. PMID:25767802

  6. Preeclampsia - will orphan drug status facilitate innovative biological therapies?

    Hahn, Sinuhe


    It is generally accepted that the development of novel therapies to treat pregnancy-related disorders, such as preeclampsia, is hampered by the paucity of research funding. Hence, it is with great interest to become aware of at least three novel therapeutic approaches for the treatment of this disorder: exploiting either the anticoagulant activity of antithrombin, the free radical scavenging activity of alpha-1-microglobulin, or the regenerative capacity of placenta-derived mesenchymal stem cells. As these projects are being carried out by small biotech enterprises, the question arises of how they are able to fund such undertakings. A novel strategy adopted by two of these companies is that they successfully petitioned US and EU agencies in order that preeclampsia is accepted in the register of rare or orphan diseases. This provides a number of benefits including market exclusivity, assistance with clinical trials, and dedicated funding schemes. Other strategies to supplement meager research funds, especially to test novel approaches, could be crowdfunding, a venture that relies on intimate interaction with advocacy groups. In other words, preeclampsia meets Facebook. Perhaps similar strategies can be adopted to examine novel therapies targeting either the imbalance in pro- or anti-angiogenic growth factors, complement activation, reduced levels of placenta protein 13, or excessive neutrophil activation evident in preeclampsia. PMID:25767802

  7. [Effects of intravenous injection of sulphated galactomannan on anticoagulant activity of rat plasma].

    Drozd, N N; Makarov, V A; Tolstenkov, A S; Lapikova, E S; Miftakhova, N T; Mestechkina, N M; Shcherbukhin, V D; Il'ina, A V; Varlamov, V P


    We studied anticoagulant activity in vitro and in vivo of sulfate depolymerisation galactomannan guar with the following characteristics: Man:Gal 1.64, molecular mass 127 kDa, sulfation degree 1.46. We found the ability of galactomannan-HSO3Na (GM) to increase the time of blood coagulation in the test aPTT with an increase of its concentration and to decrease velocity of chromogenic substrate on the factor Xa hydrolysis. Specific antithrombin and anti-factor Xa activities of GM were 35.8 +/- 1.8 IU/mg and 6.6 +/- 0.5 IU/mg, respectively. In biospecific electrophoresis complexes arise between GM and protamine sulfate. Intravenous injection of GM to rats prolonged plasma coagulation time in the aPTT test with dose reduction from 1 to 3 mg/kg. In rat thrombosis model a dose of 3 Mg/kg produced a 100% inhibition of blood clot. PMID:18946908

  8. [Effect on low-molecular-weight heparin obtained using a chitinolytic complex on the anticoagulant activity of plasma in rabbits and rats].

    Drozd, N N; Tolstenkov, A S; Makarov, V A; Miftakhova, N T; Bannikova, G E; Sukhanova, P P; Varlamov, V P; Vikhoreva, G E


    The anticoagulant activity of low-molecular-weight heparin with an average molecular weight of 4.7 kD (LMWH-4.7) has been studied. This derivative was prepared from unfractionated heparin with the help of chitinolytic enzyme complex from Streptomyces kurssanovii. The antithrombin activity of LMWH-4.7 (aIla activity) was 72 +/- 9 IU/mg and the activity with respect to the blood coagulation factor Xa (aXa activity) was 200 +/- 33 IU/mg, which corresponded to an aXa/aIIa ratio of 2.8 (necessary for effective antithrombotic drugs). The aIIa and aXa activity exhibited a dose-dependent variation upon intravenous and subcutaneous injections in rabbits, so that a high aIIa/aXa ratio was retained: 5 min after the intravenous injection of a minimum dose (0.3 mg/kg), this ratio was 2, 7, and for a greater dose (3.0 mg/kg) it reached 3.8. Subcutaneous injections were followed by slow elimination of the anticoagulant within 24 h. LMWH-4.7 upon intraperitoneal injections produced a dose-dependent inhibition of a model thrombosis in rats. Complete inhibition was observed for a dose of 3 mg/kg. Thus, it is possible to obtain active LMW heparin with the help of chitinases. PMID:17523450

  9. Sulodexide: implicazioni cliniche ed economiche

    Orietta Zaniolo


    Full Text Available Sulodexide is a highly purified glycosaminoglycan approved for leg ulcers treatment. It contains two principal components: heparan sulfate, a fast-moving heparin fraction, (80% and dermatan sulfate (20%. Sulodexide is available as an oral agent and as an injectable preparation. Its pharmacological action is obtained by dose-dependent coagulation factors inhibition: dermatan sulfate upgrades the physiological action of a selective thrombin inhibitor, heparin cofactor II, and heparan sulfate depresses activated factor X, via an increase of antithrombin III action. The antithrombotic action is enhanced by platelet aggregation inhibition and by the activation of the fibrinolytic system. This paper summarizes the results of some of the main trials that evaluated sulodexide in the treatment of peripheral occlusive arterial disease and venous leg ulcers; a trial on prevention of recurrent deep venous thrombosis with sulodexide is also reviewed. We analyzed data about the clinical and economical impact of chronic venous insufficiency with a particular attention to the cost of medication, hospitalization and management of leg ulcers. The hypothetical savings correlated to the reduction of leg ulcers incidence and healing time attainable with sulodexide have been estimated. A comparison between the different acquisition costs of the drugs frequently used to treat leg ulcers is also provided. Finally we reviewed some quality of life trials in which the psychological and sociological influence of the disease and its treatments on the patient are assessed.

  10. Characterization of IXINITY® (Trenonacog Alfa), a Recombinant Factor IX with Primary Sequence Corresponding to the Threonine-148 Polymorph.

    Monroe, Dougald M; Jenny, Richard J; Van Cott, Kevin E; Buhay, Shelly; Saward, Laura L


    The goal of these studies was to extensively characterize the first recombinant FIX therapeutic corresponding to the threonine-148 (Thr-148) polymorph, IXINITY (trenonacog alfa [coagulation factor IX (recombinant)]). Gel electrophoresis, circular dichroism, and gel filtration were used to determine purity and confirm structure. Chromatographic and mass spectrometry techniques were used to identify and quantify posttranslational modifications. Activity was assessed as the ability to activate factor X (FX) both with and without factor VIIIa (FVIIIa) and in a standard clotting assay. All results were consistent across multiple lots. Trenonacog alfa migrated as a single band on Coomassie-stained gels; activity assays were normal and showed factor IX (FIXa) per IU of FIX. The molecule has >97%  γ-carboxylation and underwent the appropriate structural change upon binding calcium ions. Trenonacog alfa was activated normally with factor XIa (FXIa); once activated it bound to FVIIIa and FXa. When activated to FIXa, it was inhibited efficiently by antithrombin. Glycosylation patterns were similar to plasma-derived FIX with sialic acid content consistent with the literature reports of good pharmacokinetic performance. These studies have shown that trenonacog alfa is a highly pure product with a primary sequence and posttranslational modifications consistent with the common Thr-148 polymorphism of plasma-derived FIX. PMID:26997955

  11. Characterization of IXINITY® (Trenonacog Alfa, a Recombinant Factor IX with Primary Sequence Corresponding to the Threonine-148 Polymorph

    Dougald M. Monroe


    Full Text Available The goal of these studies was to extensively characterize the first recombinant FIX therapeutic corresponding to the threonine-148 (Thr-148 polymorph, IXINITY (trenonacog alfa [coagulation factor IX (recombinant]. Gel electrophoresis, circular dichroism, and gel filtration were used to determine purity and confirm structure. Chromatographic and mass spectrometry techniques were used to identify and quantify posttranslational modifications. Activity was assessed as the ability to activate factor X (FX both with and without factor VIIIa (FVIIIa and in a standard clotting assay. All results were consistent across multiple lots. Trenonacog alfa migrated as a single band on Coomassie-stained gels; activity assays were normal and showed 97%  γ-carboxylation and underwent the appropriate structural change upon binding calcium ions. Trenonacog alfa was activated normally with factor XIa (FXIa; once activated it bound to FVIIIa and FXa. When activated to FIXa, it was inhibited efficiently by antithrombin. Glycosylation patterns were similar to plasma-derived FIX with sialic acid content consistent with the literature reports of good pharmacokinetic performance. These studies have shown that trenonacog alfa is a highly pure product with a primary sequence and posttranslational modifications consistent with the common Thr-148 polymorphism of plasma-derived FIX.

  12. Effects of carboxymethyl chitosan on the blood system of rats

    Highlights: → We report, for the first time, the safety of carboxymethyl chitosan in blood system. → CM-Chitosan has no significant effects on coagulation function of rats. → CM-Chitosan has no significant effects on anticoagulation performance of rats. → CM-Chitosan has no significant effects on fibrinolytic function of rats. → CM-Chitosan has no significant effects on hemorheology of rats. -- Abstract: Carboxymethyl chitosan (CM-chitosan), a derivative of chitosan, was extensively studied in the biomedical materials field for its beneficial biological properties of hemostasis and stimulation of healing. However, studies examining the safety of CM-chitosan in the blood system are lacking. In this study CM-chitosan was implanted into the abdominal cavity of rats to determine blood indexes at different times and to evaluate the effects of CM-chitosan on the blood system of rats. Coagulation function was reflected by thrombin time (TT), prothrombin time (PT), activated partial thromboplatin time (APTT), fibrinogen (FIB) and platelet factor 4 (PF4) indexes; anti-coagulation performance was assessed by the index of antithrombinIII (ATIII); fibrinolytic function was reflected by plasminogen (PLG) and fibrin degradation product (FDP) indexes; and blood viscosity (BV) and plasma viscosity (PV) indexes reflected hemorheology. Results showed that CM-chitosan has no significant effects on the blood system of rats, and provides experimental basis for CM-chitosan to be applied in the field of biomedical materials.

  13. [Interaction of hypoxia and haemostasis--hypoxia as a prothrombotic factor at high altitude?].

    Schobersberger, Wolfgang; Hoffmann, Georg; Gunga, Hanns-Christian


    For an extended period of time various research projects have been conducted on the relationship of hypoxia and haemostasis. The enclosed article contains the conclusion to which extent lack of oxygen can activate the coagulation system and induce a prothrombotic state. The majority of studies proved a shortening of coagulation times during acute exposure to hypoxia, whereas activated parameters of coagulation and fibrinolysis like prothrombin fragment F1+2 as well as thrombin-antithrombin III complexes and D-dimer remained mostly unmodified. It is suggested that a prolonged sojourn at high altitudes could lead to activation of the coagulation system through an increase of haematocrit and blood viscosity. Recently it was proven that people living at high altitudes show an enhanced risk of stroke incidents. The significance of the change in haemostasis on that outcome has not yet been part of the research. However, it has been proven that the activity of the coagulation system does not play a pathophysiological part in the development of acute mountain sickness and high altitude pulmonary edema. Recent studies also demonstrated that moderate hypoxia during long haul flights may not be the main trigger in inducing deep vein thrombosis in passengers. PMID:15966261

  14. Transcriptional modules related to hepatocellular carcinoma survival: coexpression network analysis.

    Xu, Xinsen; Zhou, Yanyan; Miao, Runchen; Chen, Wei; Qu, Kai; Pang, Qing; Liu, Chang


    We performed weighted gene coexpression network analysis (WGCNA) to gain insights into the molecular aspects of hepatocellular carcinoma (HCC). Raw microarray datasets (including 488 samples) were downloaded from the Gene Expression Omnibus (GEO) website. Data were normalized using the RMA algorithm. We utilized the WGCNA to identify the coexpressed genes (modules) after non-specific filtering. Correlation and survival analyses were conducted using the modules, and gene ontology (GO) enrichment was applied to explore the possible mechanisms. Eight distinct modules were identified by the WGCNA. Pink and red modules were associated with liver function, whereas turquoise and black modules were inversely correlated with tumor staging. Poor outcomes were found in the low expression group in the turquoise module and in the high expression group in the red module. In addition, GO enrichment analysis suggested that inflammation, immune, virus-related, and interferon-mediated pathways were enriched in the turquoise module. Several potential biomarkers, such as cyclin-dependent kinase 1 (CDK1), topoisomerase 2α (TOP2A), and serpin peptidase inhibitor clade C (antithrombin) member 1 (SERPINC1), were also identified. In conclusion, gene signatures identified from the genome-based assays could contribute to HCC stratification. WGCNA was able to identify significant groups of genes associated with cancer prognosis. PMID:27052251

  15. The role of biochemical risk factors in the etiology of AIS in children and adults.

    Kopyta, Ilona; Zimny, Mikołaj; Sarecka-Hujar, Beata


    Stroke is an abrupt onset of both focal and global neurological deficits secondary to a vascular event lasting more than 24 h and with a vascular background as its only cause. It can be triggered by a rupture of a blood vessel, aneurysm (hemorrhagic stroke, HS), thrombosis or embolisms (ischemic stroke, IS). In developed countries, it is the third most common cause of death in the adult population. Stroke in children is a rare disorder with a reported frequency of about 3 cases per 100,000 children per year. The history of acute brain ischemia is burdened with neurological complications such as motor impairment, speech impairment and intellectual delay. Moreover, in children after AIS seizures and epilepsy are also quite common. Stroke is a heterogeneous disorder; its risk factors in adults are well known, however, in pediatrics, in more than 20% cases, the cause of stroke is impossible to determine. Due to the fact that stroke usually arises as a consequence of the cerebral thrombosis, many of the mechanisms responsible for its occurrence can be considered as risk factors. We have reviewed the recent case-control studies conducted on pediatric patients regarding biochemical risk factors such as elevated levels of homocysteine, fibrinogen, protein C, protein S, antithrombin III, lipoprotein(a), cholesterol and its fractions, and compared them with the results obtained from adult patients. PMID:25428197

  16. Biopolymer-modified graphite oxide nanocomposite films based on benzalkonium chloride-heparin intercalated in graphite oxide

    Meng Na; Zhou Ninglin; Shen Jian [Jiangsu Engineering Research Center for Biomedical Function Materials, Nanjing Normal University, Nanjing 210046 (China); Zhang Shuangquan, E-mail:, E-mail:, E-mail: [Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046 (China)


    Heparin is a potent anticoagulant agent that interacts strongly with antithrombin III to prevent the formation of fibrin clots. In the present work, poly(dimethylsiloxane)(PDMS)/graphite oxide-benzalkonium chloride-heparin (PDMS/modified graphite oxide) nanocomposite films were obtained by the solution intercalation technique as a possible drug delivery system. The heparin-benzalkonium chloride (BAC-HEP) was intercalated into graphite oxide (GO) layers to form GO-BAC-HEP (modified graphite oxide). Nanocomposite films were characterized by XRD, SEM, TEM, ATR-FTIR and TGA. The modified graphite oxide was observed to be homogeneously dispersed throughout the PDMS matrix. The effect of modified graphite oxide on the mechanical properties of the nanocomposite film was investigated. When the modified graphite oxide content was lower than 0.2 wt%, the nanocomposites showed excellent mechanical properties. Furthermore, nanocomposite films become delivery systems that release heparin slowly to make the nanocomposite films blood compatible. The in vitro studies included hemocompatibility testing for effects on platelet adhesion, platelet activation, plasma recalcification profiles, and hemolysis. Results from these studies showed that the anticoagulation properties of PDMS/GO-BCA-HEP nanocomposite films were greatly superior to those for no treated PDMS. Cell culture assay indicated that PDMS/GO-BCA-HEP nanocomposite films showed enhanced cell adhesion.

  17. Studies on contact activation: effects of surface and inhibitors.

    Cameron, C L; Fisslthaler, B; Sherman, A; Reddigari, S; Silverberg, M


    Contact activation is initiated when the plasma proteins, Hageman factor (factor XII), prekallikrein and high molecular weight kininogen interact with negatively charged materials. The activation of the intrinsic pathway of blood coagulation and the production of bradykinin are among the sequelae of contact activation. The kinetics of the activation of the contact system are modified by plasma inhibitors, C1 inhibitor being quantitatively the most important. We propose that the activation of the system requires that the stimulus provided by the surface must be greater than a threshold value to overcome the effects of the inhibitors. We show in this paper that the amount of surface required for activation is much reduced in the absence of C1 inhibitor (Hereditary Angioedema) or in the cold where the inhibitor loses much of its effectiveness. Antithrombin III inhibition of activated Hageman factor is augmented by heparin which is also an activator of Hageman factor. The rate constants for inhibition remain much lower than for C1 inhibitor, however. PMID:2530427

  18. The Hemostatic System and Angiogenesis in Malignancy

    Marek Z. Wojtukiewicz


    Full Text Available Coagulopathy and angiogenesis are among the most consistent host responses associated with cancer. These two respective processes, hitherto viewed as distinct, may in fact be functionally inseparable as blood coagulation and fibrinolysis, in their own right, influence tumor angiogenesis and thereby contribute to malignant growth. In addition, tumor angiogenesis appears to be controlled through both standard and non-standard functions of such elements of the hemostatic system as tissue factor, thrombin, fibrin, plasminogen activators, plasminogen, and platelets. “Cryptic” domains can be released from hemostatic proteins through proteolytic cleavage, and act systemically as angiogenesis inhibitors (e.g., angiostatin, antiangiogenic antithrombin III aaATIII. Various components of the hemostatic system either promote or inhibit angiogenesis and likely act by changing the net angiogenic balance. However, their complex influences are far from being fully understood. Targeted pharmacological and/ or genetic inhibition of pro-angiogenic activities of the hemostatic system and exploitation of endogenous angiogenesis inhibitors of the angiostatin and aaATIII variety are under study as prospective anti-cancer treatments.

  19. Spectroscopic and Electrochemical Detection of Thrombin/5'-SH or 3'-SH Aptamer Immobilized on (porous) Gold Substrates

    Park, Buem Jin; Sa, Young Seung; Kim, Yong Hwan; Kim, Young Hun [Kwangwoon University, Seoul (Korea, Republic of)


    Thrombin is a serine protease that catalyzes the conversion of soluble fibrinogen to insoluble fibrin, and thus induces physiological and pathological blood coagulation. Therefore, it is important to detect thrombin in blood serum for purposes of diagnosis. To achieve this goal, it has been suggested that a 15-mer aptamer strongly binds with thrombin to form a G-quartet structure of the aptamer. Generally, 5'-end thiol-functionalized aptamer has been used as an anti-thrombin binder. Herein, we evaluate the possibility of utilizing a 3'-SH aptasensor for thrombin detection using SPR spectroscopy, and compare the enhancement of the electrochemical signal of the thrombin-aptamer bound on a porous gold substrate. Although the two aptamers have similar configurations, in SPR analysis, the 3'-SH aptamer was a effective aptasensor as well as 5'-SH aptamer. Results from electrochemical analysis showed that the porous gold substrate acted as a good substrate for an aptasensor and demonstrated 5-fold enhancement of current change, as compared to gold thin film.

  20. Thrombophilia

    Maurizio Zangari


    Full Text Available Thrombophilia or hypercoagulable state is a clinical condition characterized by a tendency to develop venous (less frequently arterial thrombosis. The development of a venous thromboembolic episode (VTE is the result of environmental risk factors such as age, male sex, obesity, the exposure to “risk periods” of immobilization, trauma, cancer, pregnancy or the use of exogenous hormones or antineoplastic medications often on a background of a congenital procoagulant state. Table 1 summarizes the most frequently inherited and acquired thrombophilic conditions in a population of patients with a first episode of VTE.In patients with venous thrombosis before the early nineties a biologic cause of thrombophilia was detectable in only 5% to 15% of cases and was confined to deficiencies of antithrombin, protein C, and protein S. The discovery of two prothrombotic mutations prevalent in white populations, the factor V-Arg506Gln mutation (factor V Leiden and the prothrombin G20210A mutation has significantly increased the number of patients with recognizable hereditary risk factor. The antiphospholipid antibody syndrome and elevated plasma homocysteine levels are also frequently identifiable risk factors in patients presenting with venous as well as arterial thrombosis.

  1. Syntenic assignment of human chromosome 1 homologous loci in the bovine.

    Threadgill, D S; Threadgill, D W; Moll, Y D; Weiss, J A; Zhang, N; Davey, H W; Wildeman, A G; Womack, J E


    Three mouse chromosomes (MMU 1, 3, and 4) carry homologs of human chromosome 1 (HSA 1) genes. A similar situation is found in the bovine, where five bovine chromosomes (BTA 2, 3, 5, 16, and unassigned syntenic group U25) contain homologs of HSA 1 loci. To evaluate further the syntenic relationship of HSA 1 homologs in cattle, 10 loci have been physically mapped through segregation analysis in bovine-rodent hybrid somatic cells. These loci, chosen for their location on HSA 1, are antithrombin 3 (AT3), renin (REN), complement component receptor 2 (CR2), phosphofructokinase muscle type (PFKM), Gardner-Rasheed feline sarcoma viral (v-fgr) oncogene homolog (FGR), alpha fucosidase (FUCA1), G-protein beta 1 subunit (GNB1), alpha 1A amylase, (AMY1), the neuroblastoma RAS viral (v-ras) oncogene homolog (NRAS), and alpha skeletal actin (ACTA1). AT3, REN, CR2, and GNB1 mapped to BTA 16, PFKM to BTA 5, AMY1A and NRAS to BTA 3, FGR and FUCA1 to BTA 2, and ACTA1 to BTA 28. PMID:8001974

  2. Complications of bariatric surgery: Presentation and emergency management.

    Kassir, Radwan; Debs, Tarek; Blanc, Pierre; Gugenheim, Jean; Ben Amor, Imed; Boutet, Claire; Tiffet, Olivier


    The epidemic in obesity has led to an increase in number of so called bariatric procedures. Doctors are less comfortable managing an obese patient after bariatric surgery. Peri-operative mortality is less than 1%. The specific feature in the obese patient is that the classical signs of peritoneal irritation are never present as there is no abdominal wall and therefore no guarding or rigidity. Simple post-operative tachycardia in obese patients should be taken seriously as it is a WARNING SIGNAL. The most common complication after surgery is peritonitis due to anastomotic fistula formation. This occurs typically as an early complication within the first 10 days post-operatively and has an incidence of 1-6% after gastric bypass and 3-7% after sleeve gastrectomy. Post-operative malnutrition is extremely rare after restrictive surgery (ring, sleeve gastrectomy) although may occur after malabsorbative surgery (bypass, biliary pancreatic shunt) and is due to the restriction and change in absorption. Prophylactic cholecystectomy is not routinely carried out during the same procedure as the bypass. Superior mesenteric vein thrombosis after bariatric surgery is a diagnosis which should be considered in the presence of any postoperative abdominal pain. Initially a first etiological assessment is performed (measurement of antithrombin III and of protein C and protein S, testing for activated protein C resistance). If the least doubt is present, a medical or surgical consultation should be requested with a specialist practitioner in the management of obese patients as death rates increase with delayed diagnosis. PMID:26808323

  3. Abnormal hemostatic function one year after orthotopic liver transplantation can be fully attributed to endothelial cell activation [v1; ref status: indexed,

    Freeha Arshad


    Full Text Available Background: The long-term risk of thrombotic and vascular complications is elevated in liver transplant recipients compared to the general population. Patients with cirrhosis are in a hypercoagulable status during and directly after orthotopic liver transplantation, but it is unclear whether this hypercoagulability persists over time. Aim: We aimed to investigate the hemostatic status of liver transplant recipients one year after transplantation. Methods: We prospectively collected blood samples of 15 patients with a functioning graft one year after orthotopic liver transplantation and compared the hemostatic status of these patients with that of 30 healthy individuals. Results: Patients one year after liver transplantation had significantly elevated plasma levels of von Willebrand factor (VWF. Thrombin generation, as assessed by the endogenous thrombin potential, was decreased in patients, which was associated with increased plasma levels of the natural anticoagulants antithrombin and tissue factor pathway inhibitor.  Plasma fibrinolytic potential was significantly decreased in patients and correlated inversely with levels of plasminogen activator inhibitor-1. Conclusion: One year after liver transplantation, liver graft recipients have a dysregulated hemostatic system characterised by elevation of plasma levels of endothelial-derived proteins. Increased levels of von Willebrand factor and decreased fibrinolytic potential may (in part be responsible for the increased risk for vascular disease seen in liver transplant recipients.

  4. Blood coagulation and fibrinolysis after long-duration treadmill exercise controlled by individual anaerobic threshold.

    Hilberg, Thomas; Gläser, Doreen; Reckhart, Carsten; Prasa, Dagmar; Stürzebecher, Jörg; Gabriel, Holger H W


    For rehabilitation training it is recommended that the intensity of exercise should be clearly below the individual anaerobic threshold (IAT). We investigated blood coagulation, particularly endogenous thrombin potential (ETP) and fibrinolysis following a standardized treadmill (TR) ergometer test at 90% IAT for 60-120 min. Sixteen healthy male non-smokers underwent the TR test. Blood samples were taken after a 30-min rest, immediately after exercise, and 2 h after exercise completion. Extrinsic and intrinsic total (TTP(ex+in)) and endogenous (ETP(ex+in)) thrombin potential, prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), plasmin-alpha2-antiplasmin complex (PAP), D-dimer, tissue plasminogen activator antigen and activity (tPA-AG and tPA-ACT) and plasminogen activator inhibitor type 1 antigen and activity (PAI-1-AG and PAI-1-ACT) were measured. Immediately after TR, F1+2, TAT and TTP(ex+in) were increased ( PIAT (90%) on a TR ergometer only implicates a small increase in thrombin generation markers and total (free and alpha(2)-macroglubulin-bound thrombin), but not in endogenous (free) thrombin potential alone. In contrast, fibrinolysis is distinctly increased after this type of exercise. Endurance exercise with an intensity below 90% IAT and a duration below 2 h generates a more favourable condition for fibrinolysis than for blood coagulation in healthy young subjects. Data are given as mean (SD). PMID:12883904

  5. Relationship between manic state after presenile onset and silent cerebral infarction

    Magnetic resonance (MR) imaging was performed in 12 patients who incurred manic state after the age of 50 (manic group). Twelve other age- and sex-matched patients who incurred emotional disturbance at the young age served as controls. Patients with a history of cerebral stroke and local neurologic symptoms were excluded. Silent cerebral infarction (SCI) were concomitantly seen on MR images in 66.7% in the manic group, which was significantly higher than the control group (16.7%). Approximately half of the patients in the manic group seemed to be in the parenchymal manic state associated with SCI. Patients in the manic state associated with SCI was at a risk for developing cerebral stroke. Therefore, the treatment such as antithrombin therapy for cerebrovascular disorder was considered important, from the viewpoint of early treatment of cerebral infarction. Since lesions were frequently observed in the right frontal lobe and basal nucleus in manic patients with the associated SCI, these sites may be in part involved in the occurrence of manic state. (N.K.)

  6. Platelet-derived microparticles associate with fibrin during thrombosis.

    Siljander, P; Carpen, O; Lassila, R


    Platelet-derived microparticles (MP) are reported to express both pro- and anticoagulant activities. Nevertheless, their functional significance has remained unresolved. The present study monitored the generation and fate of MP in an experimental model of thrombosis with costimulation of platelets by collagen and thrombin. When minimally anticoagulated (0.5 micromol/L PPACK) blood was perfused over immobilized fibrillar type I collagen in a flow chamber at a low shear rate (300 s(-1)), endogenous thrombin was generated, as evidenced by thrombin-antithrombin III complex. In contrast to full anticoagulation 150 micromol/L PPACK) and the absence of collagen, large platelet aggregates and fibrin ensued during perfusions over collagen in the presence of thrombin. In these thrombi, MP, defined as GPIIbIIIa- and P-selectin-positive vesicles (<1 micron), were found to align fibrin in immunofluorescence and scanning immunoelectron microscopy. Moreover, in sections of embolectomized thromboemboli from patients GPIIbIIIa- and P-selectin-positive material compatible with MP was detected in a fibrin strand-like pattern. In vitro binding studies showed that MP bound to fibrin and acted there as procoagulants. In summary, we show that MP generated during thrombus formation associate with local fibrin. This adhesive function fibrin could imply a sustained modulatory role for MP in evolving thrombi. PMID:8639834

  7. Analytical and statistical comparability of generic enoxaparins from the US market with the originator product.

    Mourier, Pierre A J; Agut, Christophe; Souaifi-Amara, Hajer; Herman, Fréderic; Viskov, Christian


    Low-molecular-weight heparins (LMWHs) are complex anticoagulant drugs, made from heparin porcine mucosa starting material. Enoxaparin sodium manufactured by Sanofi is one of the most widely prescribed LMWHs and has been used since 1993 in the USA. In 2010, US Food and Drug Administration approval for supplying generic enoxaparin was granted to Sandoz and subsequently to Amphastar. Little is known, however, of the differences in composition of these preparations. In this study, samples from several batches of generic enoxaparins were purchased on the US market and analyzed with state of the art methodologies, including disaccharide building blocks quantification, nuclear magnetic resonance (NMR), and a combination of orthogonal separation techniques. Direct high-performance liquid chromatography analysis of the different enoxaparin batches revealed distinct process fingerprints associated with each manufacturer. Disaccharide building block analysis showed differences in the degree of sulfation, the presence of glycoserine derivatives, as well as in proportions of disaccharides. Results were compared by statistical approaches using multivariate analysis with a partial least squares discriminant analysis methodology. The variations were statistically significant and allowed a clear distinction to be made between the enoxaparin batches according to their manufacturer. These results were further confirmed by orthogonal analytical techniques, including NMR, which revealed compositional differences of oligosaccharides both in low- and high-affinity antithrombin fractions of enoxaparin. PMID:26280926

  8. Relative Tissue Factor Deficiency Attenuates Ventilator-Induced Coagulopathy but Does Not Protect against Ventilator-Induced Lung Injury in Mice

    Esther K. Wolthuis


    Full Text Available Preventing tissue-factor-(TF- mediated systemic coagulopathy improves outcome in models of sepsis. Preventing TF-mediated pulmonary coagulopathy could attenuate ventilator-induced lung injury (VILI. We investigated the effect of relative TF deficiency on pulmonary coagulopathy and inflammation in a murine model of VILI. Heterozygous TF knockout (TF+/− mice and their wild-type (TF+/+ littermates were sedated (controls or sedated, tracheotomized, and mechanically ventilated with either low or high tidal volumes for 5 hours. Mechanical ventilation resulted in pulmonary coagulopathy and inflammation, with more injury after mechanical ventilation with higher tidal volumes. Compared with TF+/+ mice, TF+/− mice demonstrated significantly lower pulmonary thrombin-antithrombin complex levels in both ventilation groups. There were, however, no differences in lung wet-to-dry ratio, BALF total protein levels, neutrophil influx, and lung histopathology scores between TF+/− and TF+/+ mice. Notably, pulmonary levels of cytokines were significantly higher in TF+/− as compared to TF+/+ mice. Systemic levels of cytokines were not altered by the relative absence of TF. TF deficiency is associated with decreased pulmonary coagulation independent of the ventilation strategy. However, relative TF deficiency does not reduce VILI and actually results in higher pulmonary levels of inflammatory mediators.

  9. Preventive Efficacy of Fucoidan in Rats Exposed to γ-Rays

    Fucoidan (FDN); β-1,3-glucan is a natural polysaccharide extracted from brown seaweeds, with a wide variety of biological properties, especially the anti-inflammatory, anti-coagulation and anti-oxidative effects. The objective of this work was to evaluate the radioprotective effect of FDN. The study was performed on Sprague-Dawley rats that were administered during 10 days FDN (100 mg/ kg body wt), rats exposed to 5 Gy γ-rays, rats administered FDN + γ-rays comparing with control animals. The haemostatic parameters (protein-C, antithrombin-III and tissue-plasminogen activators activities), the haematological parameters: count of blood elements (total leukocytes, neutrophils, lymphocytes) and bone marrow cells (erythroid, lymphoid and myeloid cells) and biochemical parameters: thiobarbituric acid-reactive substances (TBARS) and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSPx) were evaluated. Results showed that γ-rays provoked marked changes in haemostatic markers, haematological parameters and anti oxidative enzymes as well as TBARS level. FDN ameliorated the alteration in all tested parameters and markers induced due to exposure to γ-rays. These data demonstrate that FDN might be viewed as a non-toxic potent agent for preventing γ-rays induced cellular hazards. Conclusion, the results may facilitate the development of a new radio protective agent with low toxicity.

  10. Diagnostic uses of snake venom.

    Marsh, N A


    Snake venom toxins are invaluable for the assay of coagulation factors and for the study of haemostasis generally. Thrombin-like enzymes (SVTLE) are used for fibrinogen and fibrinogen breakdown product assays as well as detecting dysfibrinogenaemias. Since SVTLE are not inhibited by heparin, they can be used for assaying antithrombin III in samples containing heparin. Snake venom prothrombin activators are utilised in prothrombin assays, whilst Russell's viper venom (RVV) can be used to assay clotting factors V, VII, X and lupus anticoagulants (LA). Activators from the taipan, Australian brown snake and saw-scaled viper have also been used to assay LA. Protein C (PC) and activated PC (APC) resistance can be measured by means of RVV, Protac (from Southern copperhead snake venom) and STA-Staclot (from Crotalus viridis helleri) whilst von Willebrand factor can be studied with Botrocetin (Bothrops jararaca). Finally, snake venom C-type lectins and metalloproteinase disintegrins are being used to study platelet glycoprotein receptors and show great potential for use in the routine coagulation laboratory. PMID:11910187