• Archives of Pharmacal Research
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• v.28 no.9
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• pp.1037-1041
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• 2005

Geranium (Pelargonium inquinans Ait) leaves were extracted with $80\%$ MeOH, and partitioned into n-hexane, ethyl acetate, BuOH and $H_2O$ to isolate the anticoagulant principles. The EtOAc fraction was found to be the most active, and was further purified using silica and octadecylisilane column chromatography employing a bioassay-guided fractionation method. The active compound was isolated and identified as $1,2,3,4,6-pentagalloyl-\beta-D-glucopyranose$(PGG) (compound I). The isolated anticoagulant significantly prolonged the activated partial thrombin time (APTT) and thrombin time (TT) using normal human plasma. One microgram of $1,2,3,4,6-pentagalloyl-\beta-D-glucopyranose$ showed 0.063 heparin units in the APTT and 2.73 heparin units in the TT for anti-thrombosis. This is the first report of the isolation of PGG from geranium plants.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.90-91
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• 2000

The physiological systems that control blood fluidity are both complex and elegant. Blood must remain fluid within the vasculature and yet clot quickly when exposed to nonendothelial surfaces at sites of vascular injury. There are two principle mechanisms to control a delicate balance in higher organisms (Davie & Ratnoff, 1964). Present evidence suggests that the intrinsic pathway play an important role in the growth and maintenance of fibrin formation in the coagulation cascade while a second overlapping mechanism, called the extrinsic pathway, is critical in the initiation of fibrin formation. Coagulation factors is in two mechanisms, and in order to clot blood, they are activated by a cooperation with $Ca^{2+}$, phospholipid and vitamin K etc. For example, the human placental anticoagulant protein (PAP of PAP- I), which is a $Ca^{2+}$ -dependent phospholipid binding protein (Funakoshi et al., 1987) inhibited the activity of factor Xa, so that it prolonged fibrin formation. We wondered whether any other protein was involved in regulation of the coagulant system as an anticoagulant protein from natural organisms. Natural agents would have not harmful side-effects in comparision with chemically synthesized materials such as warfarin, aspirin, phenindione, etc.. But anticoagulant agents from natural, especially marine organisms have hardly been researched except for polysaccharides from marine algae. (omitted)

• Biomolecules & Therapeutics
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• v.1 no.2
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• pp.166-170
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• 1993

Hirudin is a potent inhibitor of thrombin, which was originally obtained from the medicinal leech (Hirudo medicinalis) Now it is being produced through the recombinant technology on a large scale. Recombinant hirudin has been assayed for the anticoagulant activity by the measurement of clotting time and the inhibition of thrombin actvity using a chromogenic substrate. The assay range of partial thromboplastin time and thrombin time is within $0.2{\sim}1.0 {\mu}g/mι.$ Thrombin time is more sensitive to the measurement of clot. Ex vivo study showed the level of hirudin in rat plasma was highest in 10 min and then it was eliminated slowly. The half-life of r-hirudin was 80~110 min depending on the assay methods. Intraveneous injection of russel viper venom was used for thrombus induction combined with vents cava ligation. Inhibition of venous thrombosis was observed with i.v. hirudin. It was dependent on the concentration of hirudin.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.2
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• pp.136-141
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• 2009

Biological activities of enzymatic hydrolysate of spirulina (EHS) were investigated. EHS showed no significant effects on the growth-stimulating activity for lactic-acid bacteria and antioxidant activity. EHS showed slight in vitro growth-inhibitory effects (15% at 1.42 mg/L) on a human cervical cancer cell line (HeLa). In addition, the anticoagulant activities of EHS were measured based on three different pathways: common, intrinsic and extrinsic pathways. As an indication of anticoagulant activity on common pathway, thrombin time (TT) of EHS (100 mg/L) was measured as 155.6 sec. Activated partial thromboplastin time (aPTT) for intrinsic pathway of EHS (1,000 mg/L) was measured as 95.8 sec. Prothrombin time (PT) based on extrinsic pathway of EHS (1,000 mg/L) was measured as 10.6 sec. These data showed that EHS have influences on anticoagulant factors of common pathway and intrinsic pathway. Consequently it was found that EHS could be used as a functional food for blood circulation.

• Proceedings of the Korean Aquaculture Society Conference
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• 2006.05a
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• pp.377-378
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• 2006

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.369-375
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• 1997

We have isolated an anticoagulant polysaccharide from the alkali extracts of Coriolus versicolor. The anticoagulant polysaccharide was purified through a gradual ethanol precipitation and three concecutive chromatography of DEAE-Toyopearl 650C, Sephadex G-100, and Sepharose CL-6B by measuring activated partial thromboplastin time (aPTT). The anticoagulant polysaccharide showed the homogenecity on HPLC using a gel permeation column and had about $7.2{\times}10^{5}$ molecular weight. The polysaccharide consisted of fucose, glucose, and galactose in a molar ratio of 1.0:0.2:0.2:0.1, and also compromised 19.32% of sulfate at its constituent sugars. The polysaccharide showed the two typical bands of C-O-S $(823\;cm^{-1})$ and S=O $(1257\;cm^{-1})$ in the IR spectroscopy. The sulfated polysaccharide (CV-40-Va-1) inhibited the blood coagulation via the intrinsic pathway like heparin whose activity produced a concentration dependent effect in aPTT and thrombin time (TT).

• Journal of Microbiology and Biotechnology
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• v.21 no.10
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• pp.1036-1042
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• 2011

The exopolysaccharide botryosphaeran ($EPS_{GLC}$; a ($1{\rightarrow}3$)($1{\rightarrow}6$)-${\beta}$-D-glucan from Botryosphaeria rhodina MAMB-05) was sulfonated to produce a water-soluble fraction ($EPS_{GLC}$-S) using pyridine and chlorosulfonic acid in formamid. This procedure was then repeated twice to produce another fraction ($EPS_{GLC}$-RS) with a higher degree of substitution (DS, 1.64). The purity of each botryosphaeran sample (unsulfonated and sulfonated) was assessed by gel filtration chromatography (Sepharose CL-4B), where each polysaccharide was eluted as a single symmetrical peak. The structures of the sulfonated and re-sulfonated botryosphaerans were investigated using ultraviolet-visible (UV-Vis), Fourier-transform infrared (FT-IR), and $^{13}C$ nuclear magnetic resonance ($^{13}C$ NMR) spectroscopies. $EPS_{GLC}$ and $EPS_{GLC}$-RS were also assayed for anticoagulation activity, and $EPS_{GLC}$-RS was identified as an anticoagulant.

• Journal of Life Science
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• v.23 no.12
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• pp.1460-1470
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• 2013

To examine whether miscellaneous cereal grains have an antithrombotic effect, we investigated the anticoagulant activity of 80% ethanol extracts from eleven selected miscellaneous cereal grains. The 80% ethanol extract of hwanggeumchal sorghum (Sorghum bicolor) showed the highest anticoagulant activity, followed by that of green foxtail millet grains, in terms of thrombin time (TT). When the ethanol extract of hwanggeumchal sorghum was sequentially fractionated with n-hexane, methylene chloride, ethyl acetate, and n-butanol, the majority of the TT-inhibitory activity was detected in the hexane and methylene chloride fractions. Whereas aspirin (final conc. 480 ${\mu}g/ml$) prolonged TT by 2-fold, the ethanol extract, hexane fraction, and methylene chloride fraction in the same dose prolonged TT by 2.2-fold, 2.9-fold, and 2.5-fold, respectively. The ethanol extract of hwanggeumchal sorghum could delay activated partial thromboplastin time (APTT) as well as prothrombin time (PT). Although the APTT-inhibitory activity of the ethanol extract was mainly partitioned into the hexane and methylene chloride fractions, the PT-inhibitory activity of the ethanol extract was solely partitioned into the hexane fraction. The APTT- and PT-inhibitory activities of these organic solvent fractions were more potent than those of the control warfarin (final conc. 3.13 mg/ml). The TT-inhibitory activity of the ethanol extract was heat-stable and acid-stable. The ethanol extract, hexane fraction, and methylene chloride fraction of hwanggeumchal sorghum appeared to possess a direct fibrinolytic activity toward fibrin clotting. These results show that hwanggeumchal sorghum can exert anticoagulant and fibrinolytic effects and, thus, have the potential to be applicable as antithrombotic dietary sources.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.534-537
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• 2004

We produced twelve monoclonal antibodies(mAbs) against thyroglobulin and characterized the bindig profiles. Among them, three mAbs(TN-1, TN-2 and TN-3) were further characterized their binding specificities. TN-2 had a potent lupus anticoagulant activity and potentiated the anticoagulant effect of venom phospholipase A₂. he anticoagulant mechanism of TN-2 was elongation of the partial thromboplastin time and binding to phosphatidylserine which may have a pivot role in blood coagulation. And TN-2 was cross-reacted with ss-DNA and ds-DNA and had a characteristic of autoantibody. These results suggest that TN-2 may provide a useful tool for studying the correlation between autoimmune thyroiditis and its therapeutic effect.

• The Korea Journal of Herbology
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• v.25 no.1
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• pp.39-43
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• 2010

Objectives : This research was investigated the anticoagulant effect of the Gleditsiae spina extract. Methods : We researched prothrombin time (PT) assay, activated partial thromboplastin time (APTT) assay in vitro and in vivo using arteriovenous (A-V) shunt rat model and shortening Rat tail bleeding time (BT). A-V shunt and BT were treated with extract of Gleditsiae spina (GS) 400 mg/kg for a week. Results : Bleeding time of Gleditsiae spina extract in vivo had a significant increase as about 1.2 times and thrombus weight of Gleditsiae spina extract had a significant reduction of thrombus weight as 26%. Gleditsiae spina extract represented an effect of anticoagulation by operating on extrinsic pathway factor II, V, VII, X and intrinsic pathway factor VIII, IX, X, XI, XII in the coagulation system. Conclusions : Considering the above mentioned results, it is judged that a Gleditsiae Spina extract has a control effect of thrombus creation.