The effects of fructose diphosphate on routine coagulation tests in vitro

AbstractTo evaluate the effects of fructose diphosphate (FDP) on routine coagulation tests in vitro, we added FDP into the mixed normal plasma to obtain the final concentration of 0, 1, 2, 3, 4, 5, 6, 10, 15, 20, 25, 30 and 35 mg/mL of drug. Prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen (FBG) and thrombin time (TT) of samples were analyzed with blood coagulation analyzers from four different manufacturers(Sysmex, Stago, SEKISUI and Werfen) and their corresponding reagents, respectively. Before the experiment, we also observed whether there were significant differences in coagulation test results of different lots of reagents produced by each manufacturer. At the same time as the four routine clotting tests, the Sysmex blood coagulation analyzer and its proprietary analysis software were used to detect the change of maximum platelet aggregation rate in platelet-rich plasma after adding FDP (0, 1, 2, 3, 4, 5 and 6 mg/mL). The results of PT, aPTT and TT showed a FDP (0–35 mg/mL) concentration-dependent increase and a FBG concentration-dependent decrease. The degree of change (increase or decrease) varied depending on the assay system, with PT and aPTT being more affected by the Sysmex blood coagulation testing instrument reagent system and less affected by CEKISUI, TT less affected by CEKISUI and more affected by Stago, and FBG less affected by Stago and more affected by Sysmex. The results of PT, aPTT and TT were statistically positively correlated with their FDP concentrations, while FBG was negatively correlated. The correlation coefficients between FDP and the coagulation testing systems of Sysmex, Stago, Werfen and SEKISUI were 0.975, 0.988, 0.967, 0.986 for PT, and 0.993, 0.989, 0.990 and 0.962 for aPTT, 0.994, 0.960, 0.977 and 0.982 for TT, − 0.990, − 0.983, − 0.989 and − 0.954 for FBG, respectively. Different concentrations of FDP (0, 1, 2, 3, 4, 5 and 6 mg/mL) had different effects on the maximum aggregation rate of platelet induced by the agonists of adenosine diphosphate (ADP, 5 µmol/L), arachidonic acid (Ara, 1 mmol/L), collagen (Col, 2.5 µg/mL) and epinephrine (Epi,10 µmol/L), but the overall downward trend was consistent, that is, with the increase of FDP concentration, the platelet aggregation rate decreased significantly. Our experimental study demonstrated a possible effect of FDP on the assays of coagulation and Platelet aggregation, which may arise because the drug interferes with the coagulation and platelet aggregation detection system, or it may affect our in vivo coagulation system and Platelet aggregation function, the real mechanism of which remains to be further verified and studied.

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Influence Of Elastase On Blood Coagulation Factors: Studies In Vitro, In Rats And In Göttinger Miniatur Pigs

10.1055/s-0038-1653045 ◽

1981 ◽

Author(s):

U Kasten ◽

U Artmann ◽

T Kaethner ◽

H Burchardi ◽

H Köstering

Keyword(s):

Blood Coagulation ◽

Coagulation Factors ◽

Bleeding Complications ◽

Thrombin Time ◽

Normal Range ◽

Blood Coagulation Factors ◽

Acute Respiratory Insufficiency ◽

Coagulation Tests ◽

Antifibrinolytic Drugs

The influence of blood coagulation factors in pat. with acute respiratory insufficiency of adults, especially of the so called “pancreatitis lungs” is still unknown. In order to find out the effect of elastase, possibly activated by trypsin in pat. with acute pancreatitis, on blood coagulation factors, we performed some studies. In vitro elastase induces in plasma and blood in correlation to the dosages Enhancement of thrombingeneration in the TGT, a shortening of PTT, Thrombin time and of r- and k-time in the TEG, a loss of fibrinogen and an increase of fibrinmono-mercomplexes. In another study, elastase (960 U/ kg b.w.) was injected intravenously in rats. 30 min. later there was found a loss of fibrinogen, number of platelets, Prothrombin and a prolongation of PTT and Thrombin time and an increase of fibrinomonomercomplexes, especially in these rats, which received beside elastase Kalikreininhibitors or antifibrinolytic drugs. After repeated injections (3 times within 30 h) we found histomorpholgically thrombi as well as bleeding complications. In another study we performed (150 min) an infusion of elastase (333 U/kg b.w./h) to 9 pigs. We determined a loss of fibrinogen of platelets, of F. II, F. VII and F. XIII, a prolongation of PTT. F. VIII and F. V remained within the normal range But there was found an enhancement of Thrombin generation in the TGT, too. Compariening the results of blood coagulation tests and of histomorphological findings, elastase induced a DIC. We have to discuss their influence on ARIA and “Pancreatic lungs”.

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ORALLY ADMINISTERED VITAMIN B PROLONGS THE BLEEDING TIME AND INHIBITS PLATELET AGGREGATION IN HUMAN VOLUNTEERS.

10.1055/s-0038-1643446 ◽

1987 ◽

Author(s):

A M Randi ◽

E Sacchi ◽

M Cattaneo

Keyword(s):

Platelet Aggregation ◽

Vitamin B6 ◽

Bleeding Time ◽

Ex Vivo ◽

Vitamin B ◽

Thrombin Time ◽

Fibrin Formation ◽

Primary Hemostasis ◽

Coagulation Tests

It is known that mM concentrations of vitamin B6 inhibit human platelet aggregation and fibrin formation in vitro. There are very few and controversial data on the ex vivo effects of vitamin B6 on hemostatic parameters. We evaluated the effects of oral administration of vitamin B6 on bleeding time (BT), fibrin formation, platelet aggregation (PA) and the release reaction (RR). Vitamin B6 300 mg/day p.o., was given to 18 healthy volunteers (8 M, 10 F, aged 23-35) for 8 days. BT was measured before the first dose (Baseline), 2 hr after the first (Day 1) and the last dose (Day 8). In 7 subjects BT was measured also 7 days after the suspension of the drug (Day 15). Before and 2 hr after the first dose, prothrombin time (PT), partial thromboplastin time (PTT), thrombin time (TT), PA and the RR induced by different concentrations of ADP, PAF-acether, collagen (coll), epinephrine (epi), arachidonate (AA) were studied. BT was significantly prolonged after vitamin B6 administration, and returned to baseline values 7 days after suspension of the drug. PA and the RR induced by 1 uM ADP, 1 ug/ml coll or 5 uM epi were significantly inhibited 2 hours after vitamin B6 administration. Vitamin B6, however, did not affect PA or the RR induced by 0.2-2 uM PAF-acether, 2 uM ADP, 1 mM AA, 2 ug/ml coll, nor did it affect PT, PTT or TT. These data show that orally administered vitamin B impairs primary hemostasis, but does not affect fibrin 6 formation, as measured with standard coagulation tests.

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Anticoagulant Properties of Three Mucopolysaccharides Used in Rheumatology

Thrombosis and Haemostasis ◽

10.1055/s-0038-1665279 ◽

1983 ◽

Vol 50 (03) ◽

pp. 652-655 ◽

Cited By ~ 1

Author(s):

F Bauer ◽

P Schulz ◽

G Reber ◽

C A Bouvier

Keyword(s):

Factor Xa ◽

Degenerative Joint Disease ◽

Joint Disease ◽

Thrombin Time ◽

Coagulation Tests ◽

Amplification System ◽

Relative Potencies ◽

In Vivo Administration

SummaryThree mucopolysaccharides (MPS) used in the treatment of degenerative joint disease were compared to heparin to establish their relative potencies on 3 coagulation tests, the aPTT, the antifactor X a activity and the dilute thrombin time. One of the compounds, Arteparon®, was one fourth as potent as heparin on the aPTT, but had little or no influence on the 2 other tests. Further in vitro studies suggested that Arteparon® acted at a higher level than factor Xa generation in the intrinsic amplification system and that its effect was independent of antithrombin III. In vivo administration of Arteparon® confirmed its anticoagulant properties, which raises the question of the clinical use of this MPS.

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Effect ofToona microcarpaHarms Leaf Extract on the Coagulation System

BioMed Research International ◽

10.1155/2014/615363 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Hao Chen ◽

Min Jin ◽

Yi-Fen Wang ◽

Yong-Qing Wang ◽

Ling Meng ◽

...

Keyword(s):

Leaf Extract ◽

Factor Xa ◽

Adenosine Diphosphate ◽

Coagulation System ◽

Dependent Manner ◽

Thrombin Time ◽

Antithrombotic Agent ◽

Thrombin Activity

Toona microcarpaHarms is a tonic, antiperiodic, antirheumatic, and antithrombotic agent in China and India and an astringent and tonic for treating diarrhea, dysentery, and other intestinal infections in Indonesia. In this study, we prepared ethyl-acetate extract from the air-dried leaves ofToona microcarpaHarms and investigated the anticoagulant activitiesin vitroby performing activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) assays. Antiplatelet aggregation activity of the extract was examined using adenosine diphosphate (ADP), collagen, and thrombin as agonists, and the inhibitions of factor Xa and thrombin were also investigated. Bleeding and clotting times in mice were used to determine its anticoagulant activitiesin vivo. It is found thatToona microcarpaHarms leaf extract (TMHE) prolonged APTT, PT, and TT clotting times in a dose-dependent manner and significantly inhibited platelet aggregation induced by thrombin, but not ADP or collagen. Clotting time and bleeding time assays showed that TMHE significantly prolonged clotting and bleeding timesin vivo. In addition, at the concentration of 1 mg/mL, TMHE inhibited human thrombin activity by 73.98 ± 2.78%. This is the first report to demonstrate that THME exhibits potent anticoagulant effects, possibly via inhibition of thrombin activity.

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Blood Coagulation Disturbances During Oliguria And Polyuria Of Acute Renal Failure In Man

10.1055/s-0038-1653048 ◽

1981 ◽

Author(s):

J Schrader ◽

H Köstering ◽

H Kaiser ◽

P Kramer ◽

F Scheler

Keyword(s):

Renal Failure ◽

Acute Renal Failure ◽

Blood Coagulation ◽

Daily Basis ◽

Coagulation System ◽

Factor V ◽

Thrombin Time ◽

Complex Activity ◽

Ischemic Renal Failure ◽

Postrenal Failure

The blood coagulation system makes a significant contribution to renal damage in many disease processes. Intrarenal coagulation appears to occur in a wide variety of diseases as a primary or secondary event. As there is evidence that intraglomerular coagulation is a significant factor in the development and maintenance of oliguria in acute ischemic renal failure, blood coagulation investigations were performed in 20 patients with acute renal failure of varied etiology. The investigations were done on a daily basis from the onset of oliguria (urine flow <20 ml/h)until serum creatinine declined to less than 2,0 mg%. Thus, we were able to detect changes in blood coagulation during oliguria and polyuria. We found an enhanced thrombin generation in both oliguria and polyria. Fibrin monomer complexes were significantly increased in both states, but more predominantly in polyuria. Factor VIII and alpha-1 antitrypsin activities were also elevated. PTT and r- and k-time in TEG were shortened more in polyuria than in oliguria, whereas fibrinogen was elevated more in oliguria than in polyuria. Factor XIII activity and prothrombin complex activity (Quick’s test) were lowered in both states, the lowest values of the former being found in polyuria, the lowest values of the latter in oliguria with a normalizing tendency in the following days. Fibrinolytic activity was also decreased. No significant changes were found in plasminogen, antithrombin III, alpha-2 macroglobulin, factor V and thrombin time. In summary, we found a hypercoagulability in these patients with acute renal failure, which was more predominant during polyuria and which correlated with the tendency to thrombosis and to shorter indwelling periods of i.v. catheters in this state. Consequently, the changes in blood coagulation of 3 patients with acute postrenal failure were not as significant as those found in the other patients. The treatment with anticoagulants in patients with acute renal failure will be discussed.

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Tigecycline Interferes with Fibrinogen Polymerization Independent of Peripheral Interactions with the Coagulation System

Antibiotics ◽

10.3390/antibiotics9020084 ◽

2020 ◽

Vol 9 (2) ◽

pp. 84 ◽

Cited By ~ 1

Author(s):

Anna Brandtner ◽

Mirjam Bachler ◽

Dietmar Fries ◽

Martin Hermann ◽

Jacqueline Ruehlicke ◽

...

Keyword(s):

Qualitative Difference ◽

In Vitro Study ◽

Inhibitory Effect ◽

Coagulation System ◽

Mitochondrial Activity ◽

Hepatic Cells ◽

Spiked Plasma ◽

Fibrin Network ◽

Coagulation Tests

Tigecycline offers broad anti-bacterial coverage for critically ill patients with complicated infections. A described but less researched side effect is coagulopathy. The aim of this study was to test whether tigecycline interferes with fibrinogen polymerization by peripheral interactions. To study the effect of unmetabolized tigecycline, plasma of healthy volunteers were spiked with increasing concentrations of tigecycline. In a second experimental leg, immortalized human liver cells (HepG2) were treated with the same concentrations to test an inhibitory effect of hepatic tigecycline metabolites. Using standard coagulation tests, only the activated thromboplastin time in humane plasma was prolonged with increasing concentrations of tigecycline. Visualization of the fibrin network using confocal live microscopy demonstrated a qualitative difference in tigecycline treated experiments. Thrombelastometry and standard coagulation tests did not indicate an impairment of coagulation. Although the discrepancy between functional and immunologic fibrinogen levels increased in cell culture assays with tigecycline concentration, fibrinogen levels in spiked plasma samples did not show significant differences determined by functional versus immunologic methods. In our in vitro study, we excluded a direct effect of tigecycline in increasing concentrations on blood coagulation in healthy adults. Furthermore, we demonstrated a rapid loss of mitochondrial activity in hepatic cells with supra-therapeutic tigecycline dosages.

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Evaluation Procoagulant Activity and Mechanism of Astragalin

Molecules ◽

10.3390/molecules25010177 ◽

2020 ◽

Vol 25 (1) ◽

pp. 177 ◽

Cited By ~ 3

Author(s):

Changqin Li ◽

Miyun Hu ◽

Shengjun Jiang ◽

Zhenhua Liang ◽

Jinmei Wang ◽

...

Keyword(s):

Plasma Viscosity ◽

Procoagulant Activity ◽

Coagulation System ◽

Control Group ◽

Thrombin Time ◽

Coagulation Time ◽

Erythrocyte Sedimentation ◽

Procoagulant Effect

Astragalin, isolated from flowers of Rosa chinensis Jacq., is a kind of flavonoid, with anti-inflammatory, antioxidant, antiviral, analgesic, antibacterial, antiallergic, and antihepatotoxic effects. However, no studieson the procoagulant effect of astragalin have been reported. This study aimed to investigate the procoagulant activity of astragalin and its mechanism. Its procoagulant effect was investigated by activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin time (PT), and fibrinogen (FIB) in vitro, and a rat model established by heparin sodium was used to evaluate the mechanism for the procoagulant effect in vivo. The results showed that astragalin had good procoagulant effects compared with the control group in vitro. Compared with the model group in vivo, astragalin could shorten the coagulation time and significantly increase the number of platelets. Meanwhile, astragalin could significantly reduce the effectual time of PT and APTT and increase the content of FIB. The contents of 6-keto-PGF1α and eNOS significantly decreased. Astragalin could increase whole blood viscosity (WBV), plasma viscosity (PV), erythrocyte sedimentation rate (ESR) and packedcell volume (PCV). All of the above revealed that astragalin had good procoagulant effects by promoting the intrinsic and extrinsic coagulation system.

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Effect of Furostanol Saponins from Allium Macrostemon Bunge Bulbs on Platelet Aggregation Rate and PI3K/Akt Pathway in the Rat Model of Coronary Heart Disease

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/9107847 ◽

2019 ◽

Vol 2019 ◽

pp. 1-7

Author(s):

Hui Feng ◽

Zhipeng Wang ◽

Changsong Wang ◽

Xinyi Zhu ◽

Zhigang Liu ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Platelet Aggregation ◽

Akt Phosphorylation ◽

Aggregation Rate ◽

Furostanol Saponins ◽

Sd Rats ◽

Allium Macrostemon

Aim. To investigate the effect of Furostanol Saponins from Allium Macrostemon Bunge Bulbs (FSAMB) on platelet aggregation rate of rats with coronary heart disease and discuss the mechanism of FSAMB affecting the platelet aggregation rate through PI3K/Akt pathway. We established the rat models with coronary heart disease (CHD) and prepared the platelet-rich plasma. The effect of different concentrations of FSAMB on platelet aggregation in SD rats induced by ADP was observed in vitro and in vivo. And Lactate Dehydrogenase (LDH), Creatine Kinase-MB Form (CK-MB), and Cardiac Troponin I (cTnI) are detected in the blood to know the level of damage to heart cells. The expansion of platelets in the immobilized fibrinogen in different concentrations of FSAMB was observed. Western blot was conducted to detect the phosphorylation level of protein kinase B (also known as Akt) and the expression level of phosphoinositide 3-kinase (PI3K). We found that FSAMB had a significant inhibitory effect on the ADP-induced platelet aggregation in vitro. Intragastric administration of FSAMB also inhibited platelet aggregation induced by ADP in rats. LDH, CK-MB, and cTnI levels in serum of rats in FSAMB (672 mg/kg) group were lower than those in the model control group after the intervention (P<0.01 or P<0.05). FSAMB inhibited the expansion of platelets on immobilized fibrinogen. Also, FSAMB inhibited ADP-induced platelet PI3K expression and Akt phosphorylation. The inhibition of Akt phosphorylation by FSAMB was more obvious after the inhibition of the expression of PI3K. This study demonstrated that FSAMB can reduce the degree of myocardial cell damage and inhibit ADP-induced platelet aggregation in SD rats, possibly by inhibiting platelet PI3K/Akt signaling pathway in vitro and in vivo.

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Effect of capsaicin and dihydrocapsaicin on in vitro blood coagulation and platelet aggregation

Thrombosis Research ◽

10.1016/j.thromres.2009.05.001 ◽

2009 ◽

Vol 124 (6) ◽

pp. 721-723 ◽

Cited By ~ 31

Author(s):

Murray J. Adams ◽

Kiran D.K. Ahuja ◽

Dominic P. Geraghty

Keyword(s):

Platelet Aggregation ◽

Blood Coagulation

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In Vitro Effect of Danaparoid Sodium (Orgaran®) on Thrombin Generation after Minimal Tissue Factor Pathway Activation.

Blood ◽

10.1182/blood.v106.11.4151.4151 ◽

2005 ◽

Vol 106 (11) ◽

pp. 4151-4151

Author(s):

Ismail Elalamy ◽

Anna D. Petropoulou ◽

Mohamed Hatmi ◽

Meyer M. Samama ◽

Grigoris T. Gerotziafas

Keyword(s):

Molecular Weight ◽

Tissue Factor ◽

Thrombin Generation ◽

Lag Time ◽

Dependent Manner ◽

Pathway Activation ◽

Coagulation Tests ◽

Vitro Effect ◽

Routine Coagulation

Abstract Introduction: Orgaran® (Org 10172) is a low molecular weight heparinoid which consists of natural sulphated glycosaminoglycans (heparan, dermatan, chondroitin sulphate). It has a mean molecular weight of approximately 6 kDa (4–10 kDa), an excellent bioavailability following subcutaneous administration and an anti-Xa/anti-IIa activity ratio superior to 22. It is the anticoagulant of choice in patients developping Heparin-Induced Thrombocytopenia (HIT), whereas its’ use is also proposed for surgical thromboprophylaxis. Orgaran® has no effect on routine coagulation tests (aPTT, PT, TT). Thrombin generation test(TG) is a global clotting assay proven to be sensitive to the anticoagulant effect of LMWHs and specific FXa inhibitors (i.e. fondaparinux and BAY-597939). In this in vitro study, we determined the tissue factor (TF)-induced TG inhibition potency of Orgaran® using the Thrombogram-Thrombinoscope® assay. Materials and Methods: TG was assessed after TF pathway activation in Platelet Rich Plasma (PRP) (1.5x105 platelets/μl) using diluted thromboplastin (Dade Innovin®, 1:1000 final dilution). The clotting process is provoked by a physiologically relevant TF concentration. Orgaran® was added to control plasma from 8 healthy volunteers at five different final concentrations (0.2, 0.4, 0.6, 0.8 and 1IU anti-Xa/ml). TG was initiated by adding the triggering solution containing CaCl2 and the fluorogenic substrate. The analyzed TG parameters are the lag time, the maximal concentration of thrombin (Cmax), the time to reach Cmax (Tmax), the TG velocity and the endogenous thrombin potential (ETP). Results: Orgaran® prolonged significantly the lag time and the Tmax at a concentration over 0.40 IU anti-Xa/ml (p<0.05). At the lowest studied concentration (0.20 IU anti-Xa/ml), lag time and Tmax were only prolonged by 12%, whereas their maximal prolongation (around 50%) was observed at 1IU anti-Xa/ml. Furthermore, Orgaran® inhibited ETP, Cmax and TG velocity in an almost linear dose dependent manner. A significant inhibition of ETP, Cmax and TG velocity was obtained at concentrations superior to 0.20 IU anti-Xa/ml. (p<0.05). At the highest studied concentration (1IU anti-Xa/ml) Orgaran® suppressed all TG parameters by about 80% (Table 1). Conclusion: Orgaran® exhibited a significant inhibitory activity of in vitro TG. At concentrations achieved in clinical practice (prophylactic or therapeutic dose), Orgaran® modified in vitro TG profile while it has no effect on routine coagulation tests. Thus, TG assay is a sensitive method for monitoring Orgaran® and this test requires a clinical prospective evaluation. Table 1. Determination of IC20 and IC50 anti-Xa inhibitory concentrations of Orgaran® on TG parameters Lag Time Tmax ETP Cmax Velocity IC: Inhibitory Concentration * or Concentration increasing 20% and 50% the lag time and the Tmax respectively IC 20 (IU/ml) 0.30 0.30 0.18 0.18 0.15 IC 50 (IU/ml) 0.83 >1 0.30 0.50 0.35 1IU anti-Xa/ml 53% 47% 68% 76% 84%

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