Effect of i. v. and Oral Administration of Heparinoids G31150, G31150-A and of Nitrilotriacetic Acid on Blood Coagulation

SummaryThe heparinoid G31150-A and nitrilotriacetic acid (NTA) were compared with heparinoid G31150 and heparin. In vitro the anticoagulant activity of NTA was 1/20000, G31150-A 1/8th and G31150, 1/10th of heparin. On intravenous injection in dogs, the degree and duration of hypocoagulability was about the same for the two heparinoids but less than predicted from the effects in vitro. Injection of 25 mg NTA/kg caused a slight increase in coagulation time lasting over 4 hours. When heparinoid G31150-A and NTA were given orally, there was an increase in clotting time of blood and metachromatic activity appeared in the urine. The greatest amount of activity was excreted with the highest dose of NTA alone. Microelectrophoresis of the urinary concentrate indicated the excreted material after NTA was not heparin, but another mucopolysaccharide; after G31150, a heparinoid. These results were confirmed by the resulting absorption spectrum of the dye obtained with added urinary concentrate. It is suggested that the effects produced when a chelating agent is given with oral heparin and heparinoids may be due to the release of a mucopolysaccharide.

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Anticoagulant Activity of Dayak Onion Bulb (Eleutherine bulbosa) Extract on Human Blood Samples

Health Notions ◽

10.33846/hn50202 ◽

2021 ◽

Vol 5 (2) ◽

pp. 46-48

Author(s):

Anik Handayati ◽

Syamsul Arifin ◽

Monica Putji Astuti

Keyword(s):

Blood Coagulation ◽

Human Blood ◽

Anticoagulant Activity ◽

Onion Bulb ◽

Clotting Time ◽

Coagulation Time ◽

Blood Samples ◽

Vein Thrombosis ◽

Onion Bulbs ◽

Human Blood Samples

Cardiovascular disease can occur due to disruption of the hemostatic system by forming blood clots in blood vessels, or commonly known as deep vein thrombosis. The curative action that has been carried out in the form of heparin anticoagulant administration had many side effects such as bleeding, hematuria, thrombocytopenia, and hypersensitivity; therefore, alternative anticoagulant ingredients such as onion bulbs have emerged. Dayak onion bulb (Eleutherine bulbosa) is known to contain eleutherinol compound, which has anticoagulant activity. This study aims to determine the potential anticoagulant activity of Dayak onion bulbs extract (Eleutherine bulbosa) on human blood samples. This anticoagulant activity test was carried out by observing the length of time for blood clotting in the extract treatment group with several concentrations (0, 10, 12.5, 16, 25, and 50%) by Clotting Time (Lee-White) method. The results showed a significant delay in blood coagulation time in the group treated with Dayak onion bulbs extract as increasing the concentration. About 50% Dayak onion onion bulb extract was showed a twofold increase in blood coagulation time thanthat of no extract administration. So that, Dayak bulb onion has the opportunity to be an alternative natural anticoagulant. Keywords: anticoagulants; Dayak onion bulbs (Eleutherine bulbosa); clotting time

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Effect of Temperature, Velocity Gradient and I.V. Heparin on in Vitro Blood Coagulation and Platelet Aggregation

Thrombosis and Haemostasis ◽

10.1055/s-0038-1654086 ◽

1967 ◽

Vol 17 (01/02) ◽

pp. 112-119 ◽

Cited By ~ 3

Author(s):

L Dintenfass ◽

M. C Rozenberg

Keyword(s):

Blood Coagulation ◽

Velocity Gradient ◽

Elevated Temperatures ◽

Morphological Changes ◽

Effect Of Temperature ◽

Clotting Time ◽

Progressive Change ◽

Aggregation Of Platelets ◽

Microscopic Techniques

SummaryA study of blood coagulation was carried out by observing changes in the blood viscosity of blood coagulating in the cone-in-cone viscometer. The clots were investigated by microscopic techniques.Immediately after blood is obtained by venepuncture, viscosity of blood remains constant for a certain “latent” period. The duration of this period depends not only on the intrinsic properties of the blood sample, but also on temperature and rate of shear used during blood storage. An increase of temperature decreases the clotting time ; also, an increase in the rate of shear decreases the clotting time.It is confirmed that morphological changes take place in blood coagula as a function of the velocity gradient at which such coagulation takes place. There is a progressive change from the red clot to white thrombus as the rates of shear increase. Aggregation of platelets increases as the rate of shear increases.This pattern is maintained with changes of temperature, although aggregation of platelets appears to be increased at elevated temperatures.Intravenously added heparin affects the clotting time and the aggregation of platelets in in vitro coagulation.

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Mechanism of Heparin Rebound

10.1055/s-0038-1665846 ◽

1979 ◽

Author(s):

I. Fabian ◽

M. Aronson

Keyword(s):

Clotting Time ◽

Coagulation Time ◽

Working Hypothesis ◽

Plasma Enzymes

Heparin rebound is occasionally encountered following protamin sulfate administration for the neutralization of excess of heparin. in these situations the anticoagulatory properties of heparin are initially abolished, but within several hours the blood display again an increased clotting time. The purpose of this work was to try to reproduce the phenomenon under in vitro conditions, and to provide a working hypothesis for its explanation. Under the condition used the following parameters were obtained (according to the APTT method): clotting time of untreated plasma 50-55 seconds; with the addition of 4 units heparin/ml plasma>3 minutes; and with the addition of 50-100 μg of protamin to the heparinized plasma the clotting time reverts to 50-55 seconds. It was, however, found that incubation of heparin-protamin complex with the plasma at 37° for several hours, reduced the effectivity of the protamin, in other words, a longer coagulation time was observed. Subsequently, we found by electrophoretical methods that (heparin bound) protamin is proteolitically degraded upon incubation in plasma, the anticoagulatory properties of the heparin remaining intact. in summary, our results are compatible with the hypothesis that heparin rebound can be explained by the degradation of protamin by plasma enzymes and the release of this newly available heparin Into the circulation. The importance of this phenomenon in conjunction with other observations previously described by us are discussed.

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Evaluation of anticoagulant activity of aqueous extract of Cestrum nocturnum

International Journal of Phytomedicine ◽

10.5138/09750185.1969 ◽

2017 ◽

Vol 8 (4) ◽

pp. 525

Author(s):

Chandra Kishore Tyagi ◽

Deenanath Jhade ◽

Sunil Kumar Shah

Keyword(s):

Aqueous Extract ◽

Anticoagulant Activity ◽

Ethanolic Extract ◽

Dependent Manner ◽

Thrombin Time ◽

Clotting Time ◽

Standard Procedures ◽

Prolonged Aptt ◽

Pharmacologically Active

The study evaluated anticoagulant properties of the aqueous extract of Cestrum nocturnum using aPTT-Activated Partial Thromboplastin Time, PT- Prothrombin Time & TT-Thrombin Time as standard procedures.For in vitro coagulation assays, aqueous extract of plant prolonged APTT, TT, and PT clotting times in a dose-dependent manner (Table 7). It prolonged APTT clotting time from 45 ± 2 (2mg/mL) to 82.2 ± 2.63s (10mg/mL), PT clotting time from 20.4 ± 1.49 (2mg/mL) to 31.4 ± 2.15s (10mg/mL), and TT clotting time from 9.2 ± 1.16 (2mg/mL) to 17.4 ± 1.01s (10mg/mL) at the concentration of 2 to 10mg/mL. Heparin prolonged APTT and PT clotting times more than 111.8s and 40.8s, respectively, at a concentration of 1 IU/mL. Heparin prolonged TT clotting times more than 20.6s at a concentration of 1 IU/mL.The phytochemical screening of the plant confirm the presence of saponin in the water and ethanolic extract, Alkaloid in all the extract except hexane extract, tannin in water, ethanol and methanol extract, amino acid in water and ethanolic extract, carbohydrate in water and methanolic extract and triterpenoids and glycoside were absent in all the extracts. The results demonstrated that the aqueous extract of Cestrum nocturnum possesses pharmacologically active anticoagulant principles that could be isolated and evaluated for clinical or physiological purposes.

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A Comparison of Anticoagulation Activity of a Potent Heparin Preparation in Different Test

10.1055/s-0039-1687438 ◽

1979 ◽

Author(s):

A.S. Bhargava ◽

J. Heinick ◽

Chr. Schöbel ◽

P. Günzel

Keyword(s):

Blood Clotting ◽

Anticoagulant Activity ◽

Factor Xa ◽

Thrombin Time ◽

Beagle Dogs ◽

Clotting Time ◽

Relative Potencies ◽

Heparin Preparation

The anticoagulant effect of a new potent heparin preparation was compared with a commercially available heparin in vivo after intravenous application in beagle dogs. The anticoagulant activity was determined using thrombin time, activated partial thromboplastin time and whole blood clotting time after 5, 10 and 30 minutes of application. The relative potency of the new heparin preparation (Scherinq) was found to be 1.62 to 2.52 times higher than heparin used for comparison (150 USP units/mg, Dio-synth). The anticoagulant properties of both preparations were also studied in vitro using dog and human plasma. The relative potencies in vitro correlated well with those obtained in vivo. Further characterization with amidolytic method using chromogenic substrate for factor Xa and thrombin (S-2222 and S-2238 from KABI, Stockholm) showed that heparin (Schering) contains 243 to 378 USP units/raq depending upon the test systems used to assay the anticoagulation activity and in addition, proves the validity of the amidolytic method.

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THE RELATIONSHIP BETWEEN HEPARIN DOSAGE AND CLOTTING TIME

Blood ◽

10.1182/blood.v3.10.1197.1197 ◽

1948 ◽

Vol 3 (10) ◽

pp. 1197-1212 ◽

Cited By ~ 25

Author(s):

L. B. JAQUES ◽

ANN G. RICKER

Keyword(s):

Anticoagulant Activity ◽

The Body ◽

Time Response ◽

Fixed Dose ◽

Clotting Time ◽

Anticoagulant Action ◽

India Ink ◽

The Relationship ◽

Heparin Dosage

Abstract 1. The relationship between clotting time and heparin dosage has been studied in the dog. 2. On the addition of heparin to blood in vitro, a linear relation is found between heparin dosage and the logarithm of the clotting time obtained. The sensitivity of the blood sample to the action of added heparin is influenced both by the individual (coagulability of the blood before withdrawal) and by the technics of withdrawal and of determination of the clotting time. It is indicated that alterations in the latter may be used to extend the range of measurable hypocoagulability due to heparin. Incubation of heparin with blood for ten minutes increases its anticoagulant effect. 3. When moderate doses of heparin are injected intravenously, five to fifteen minutes are required for the clotting time to reach a maximum. No evidence of a biphasic response was obtained. The maximum clotting time obtained is greater than it is with the same amount of heparin added to the blood in vitro, due to the effect of incubation of heparin with blood on its anticoagulant activity. The in- terval required for the clotting time to return to normal is quite short, and with a given dosage is constant with different animals. Factors influencing the relation between duration of hypocoagulability and dosage are discussed. 4. A test has been devised to determine the sensitivity of the animal to the anticoagulant action of heparin. The clotting time response to certain concentrations of heparin added to the blood in vitro is determined. A fixed dose of heparin is then injected intravenously and the clotting time response is again determined. The response in vitro measures the sensitivity of the clotting system to heparin, while the in vivo response, when interpreted in the light of the in vitro response, measures the ability of the body to remove heparin from the circulation. 5. By means of this test, it has been determined that anesthesia with pentobarbital decreased the coagulability of the blood, urethane had no effect on coagulability, while the effect of ether was variable. The injection of india ink and evisceration caused a hypercoagulability, while removal of the kidneys had little effect. 6. When the sensitivity of the blood to the anticoagulant action of heparin was tested during these procedures, pentobarbital and nephrectomy had no effect, ether caused an increase in sensitivity, urethane a decrease. The injection of india ink and also evisceration markedly decreased the sensitivity of the blood to the anticoagulant action of heparin. 7. Anesthesia with pentobarbital, ether or urethane, the injection of india ink, removal of the kidneys, or removal of the gastrointestinal tract, had no effect on the duration of heparin action in the body.

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Mechanism of Heparin Rebound

10.1055/s-0039-1684574 ◽

1979 ◽

Author(s):

I. Fabian ◽

M. Aronson

Keyword(s):

Clotting Time ◽

Coagulation Time ◽

Working Hypothesis ◽

Plasma Enzymes

Heparin rebound is occasionally encountered following protamin sulfate administration for the neutralization of excess of heparin. In these situations the anticoagulatory properties of heparin are initially abolished, but within several hours the blood display again an increased clotting time. The purpose of this work was to try to reproduce the phenomenon under in vitro conditions, and to provide a working hypothesis for its explanation. Under the condition used the following parameters were obtained (according to the APTT method): clotting time of untreated plasma 50-55 seconds; with the addition of 4 units heparin/ml plasma>3 minutes; and with the addition of 50-100 μg of protamin to the heparinized plasma the clotting time reverts to 50-55 seconds. It was, however, found that incubation of heparin-protamin complex with the plasma at 37° for several hours, reduced the effectivity of the Protamin, in other words, a longer coagulation time was observed. Subsequently, we found by electrophoretical methods that (heparin bound) protamin is proteolitically degraded upon incubation in plasma, the anticoagulatory properties of the heparin remaining intact. In summary, our results are compatible with the hypothesis that heparin rebound can be explained by the degradation of Protamin by plasma enzymes and the release of this newly available heparin in to the circulation. The importance of this phenomenon in conjunction with other observations previously described by us are discussed.

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THE BLOOD COAGULATION MECHANISM OF FROGS, WITH RESPECT TO THE SPECIES SPECIFICITY OF THROMBO PLASTIN, TO INTRACARDIAL THROMBIN INJECTION, AND TO THE EFFECT OF SEASONAL CHANGES

Canadian Journal of Medical Sciences ◽

10.1139/cjms52-053 ◽

1952 ◽

Vol 30 (5) ◽

pp. 420-424

Author(s):

John J. Spitzer ◽

Judy A. Spitzer

Keyword(s):

Prothrombin Time ◽

Blood Coagulation ◽

Seasonal Changes ◽

Thrombin Injection ◽

Clotting Time ◽

Coagulation Time ◽

Coagulation Mechanism ◽

Significant Difference ◽

Whole Blood Coagulation Time ◽

Blood Coagulation Mechanism

1. The prothrombin time of frogs is much shorter using frog rather than rabbit thromboplastin in the determination. The contrary is true for dogs, rabbits, and rats. In guinea pigs there was no significant difference between the two kinds of thromboplastin. 2. The intracardiai injection of thrombin in frogs induces a triphasic response regarding the whole blood coagulation time: (a) shortening, (b) prolongation, and (c) return towards the normal. The return is quicker than in the case of mammals. 3. The following seasonal changes were obtained in “winter” frogs as compared to “summer” animals: (a) pro longed clotting time, (b) increased prothrombin time, and (c) lessened effectiveness of intracardiai thrombin solution. The clotting time with added thrombin was not changed.

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In Vitro Inhibition of Blood Coagulation by Tripeptide Aldehydes - A Retrospective Screening Study Focused on the Stable D-MePhe-Pro-Arg-H · H2SO4

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648441 ◽

1992 ◽

Vol 67 (03) ◽

pp. 325-330 ◽

Cited By ~ 17

Author(s):

Daniel Bágdy ◽

Èva Barabás ◽

Sándor Bajusz ◽

Erzsébet Széll

Keyword(s):

Blood Coagulation ◽

Plasma Proteins ◽

Anticoagulant Activity ◽

Coagulation Factors ◽

Inhibitory Effect ◽

Inhibitory Potency ◽

Peptide Aldehydes ◽

Screening Study ◽

Tissue Homogenates

SummaryA series of peptide aldehydes synthetized in our institute during the last 15 years were screened to detect their inhibitory effect on blood coagulation. Simple conventional clotting assays, platelet function tests and fibrinolytic methods were used to evaluate the inhibitory potency of the compounds in complex clotting systems as well as their supposed antifibrinolytic effect in vitro. Special attention was paid to the possible interactions with blood cells and plasma proteins, and to the functional stability of the inhibitors in several tissue homogenates. D-Phe-Pro-Arg-H (GYKI-14166, RGH-2958), Boc-D-Phe-Pro-Arg-H (GYKI-14451) and D-MePhe-Pro-Arg-H (GYKI-14766) were found to be the most potent inhibitors. The peptide aldehydes via formation of reversible complexes with thrombin impede the enzyme to react with the coagulation factors, platelet membrane and vessel wall. The compounds inhibit platelet aggregation induced by thrombin specifically without changing the sensitivity of platelets to other inducers. D-Phe-Pro-Arg-H and D-MePhe-Pro-Arg-H showed no antifibrinolytic effect. D-MePhe-Pro-Arg-H and Boc-D-Phe-Pro-Arg-H proved to be stable in dry state for years and in solution at room temperature for several days. The anticoagulant activity of the compounds was declared in NIH antithrombin units.

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Distribution of Phenylindanedione in Blood and Tissues after Oral and Intravenous Administration

Thrombosis and Haemostasis ◽

10.1055/s-0038-1656276 ◽

1958 ◽

Vol 02 (03/04) ◽

pp. 236-249 ◽

Cited By ~ 3

Author(s):

G. J Millar ◽

Mary Ogilvie Mersereau ◽

J Lowenthal ◽

L. B Jaques

Keyword(s):

Absorption Spectrum ◽

Oral Administration ◽

Intravenous Injection ◽

Optical Density ◽

Intravenous Administration ◽

Phosphate Buffer ◽

Red Cells ◽

Tissue Levels

SummaryThe absorption spectrum and Polarographie behaviour are described for phenylindanedione. The drug may be estimated by measuring the optical density at 338 nιμ or the Polarographie reduction ware in a phosphate buffer at pH 11.8. For the estimation of the drug in tissue and plasma, the substance is extracted with toluene, taken up in alkali and estimated spectrometrically. For red cells, it is necessary to extract with acetone and after evaporation, take up the residue in water and determine the phenylindanedione polarographically. Blood and tissue levels were determined after administration to rabbits and dogs. Phenylindanedione disappeared from the plasma after intravenous injection much more rapidly than dicumarol. Considerable amounts of the drug were detected in liver and intestine one hour later. On oral administration, after 4—6 hours 50°/o was still recovered from the intestine, while 10—20% was found in the liver. Phenylindanedione and / or its metabolic derivative could be detected in the erythrocytes of rabbits but not in those of dogs.

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