• Journal of Life Science
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• v.24 no.8
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• pp.873-879
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• 2014

The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed after n-butanol fractionation was subjected to purification by medium pressure and high pressure liquid chromatography. The two neolignans, 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl -3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside (a neolignan glycoside) and 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$) were identified by $^1H$ and $^{13}C$ NMR spectra. The effect of the purified compounds, the neolignan glycoside and icariside $E_4$ on thrombin inhibition were investigated by measuring thrombin clotting time in plasma. As a result, the clotting of the neolignan glycoside was delayed four times compared to that of icariside $E_4$. In addition, an analysis of the inhibition effect by changing the concentration showed that the clotting time was delayed in accordance with an increase in the concentration of the neolignan glycoside. Furthermore, we examined the interaction of thrombin and fibrinogen to clarify the action mechanism. As a result, the delay of clotting time in the response of thrombin and pure fibrinogen may indicate that neolignan glycosides inhibit the thrombin action in a direct manner, leading to the suppression of fibrin generation.

• Journal of Chest Surgery
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• v.30 no.11
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• pp.1092-1096
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• 1997

Thromboelastography(TEG) enables a global assessment of hemostatic function to be made from a single blood sample, documenting the interaction of platelets with protein coagulation cascade from the time of the initial platelet-fibrin interaction, through platelet aggregation, clot strengthening and fibrin cross linking to eventual clot Iysis. Thirty-five patients(mean age 34$\pm$ 12) undergoing open heart surgery from April 1st, 1996 to August 31th, 1996 were investigated at preoperatively and immediate, one hour, and 24 hours after cessation of cardiopulmonary bypass using TEG. Comparisons were made between classic hematological indices and TEG data. There were statistically significant correlation between maximal amplitude(MA) and platelet count before CPB, activating clotting time(ACT) and TEG date(R time, K time and a angle) at 24-hour after CPB. The data on the predictive accuracy for postoperative bleeding at 24-hour after CPB, the TEG was significantly better than ACT(57%) or the coagulation profiles(43%) as a predictor of postoperative bleeding, with an accuracy rate of 100% (P=0.0043). In conclusion, TEG seems to be easy to use, clinically accurate, cost effective and provides data which can effectively manage a patient's hemostasis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.5
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• pp.878-884
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• 1996

A bacterial strain, designated as Bacillu sp. IJ-3 strain, was shown to produce the extracellular soy protein coagulating enzyme and culture conditions for the production of enzyme by this microbial strain was investigated. The culture medium giving a maximum soy protein coagulating activity was consist of 20%(w/v) soymilk, 2.0%(w/v) glucose, 4.0%(w/v) yeast extract, 5.0%(w/v) polypeptone and 1.0%(w/v) potassium phosphate, monobasic. Initial pH was optimal at 6.0 and the enzyme activity in the culture usually reached a maximal level of fermentation at $35^{\circ}C.$ After the culture medium adjustment where required, enzyme activity was reached maximum at 72 hour of cultivation but this enzyme activity was reduced quickly. It can be assumed that Bacillu sp. IJ-3 strain enzyme has a specificity toward the 75 globulin.

• Biomedical Science Letters
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• v.3 no.2
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• pp.83-88
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• 1997

In septic patients, disseminated intravascula. coagulation (DIC) occurs frequently and is a pathologic condition associated with a variety of critical illness. DIC may complicate the already complex clinical situations and contribute to the high mortality. Nevertheless, its pathogenic mechanisms are not completely elucidated. Present study was prospectively designed to understand the pathogenetic mechanisms involved in the development of DIC. 15 rats were subjected to study and according to the aim, they were divided into three groups： group I, control (not treated-endotoxin, n=5)； group II (12 hours after endotoxin injection, n=5)； group III (24 hours after endotoxin injection, n=5). Experimental DIC was induced in rats by a bolus injection of endotoxin (1mg/kg, E. coli serotype 055:B5). Blood was collected by direct puncture of the heart. Platelet count, fibrinogen and plasminogen concentration, antithrombin III, D-dimer and complement components (C3 and C4) were measured in all subjects. In group II and III, there were apparent signs of DIC, including thrombocytopenia, decreased fibrinogen (but increase in group III), reduced C3 and antithrombin III, and elevated D-dimer. These data indicated that endotoxin might induce the activation of several pathways such as coagulation, fibrinolytic and complement cascade, causing DIC and subsequent multiple organ failures. Ultimately, the increased knowledge of the various pathogenetic mechanisms of coagulation activation and fibrinolysis in endotoxin-induced DIC may have prophylactic or therapeutic implications.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.3
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• pp.262-268
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• 2007

This study was conducted to investigate various physiochemical activities of carrageenan hydrolysates obtained with organic acid treatments. The hydrolysates treated with citrate and malate at $100^{\circ}C,\;110^{\circ}C\;and\;120^{\circ}C$ had antimicrobial activities against Bacillus cereus and Bacillus subtilis. Especially, the hydrolysates with malate at $120^{\circ}C$ for 180 min treatment had the strongest antimicrobial activity to Bacillus subtilis. Regardless of the hydrolysis conditions, inhibition ratios of tyrosinase activity by citrate and malate under $100{\sim}120^{\circ}C$ were over 97%. Especially, the inhibition ratios for tyrosinase activity of the hydrolysates obtained with citrate at $100^{\circ}C$ for 180 min and at $120^{\circ}C$ for 90 min were 99.4% and 98.2%, respectively. Also, the inhibition ratios for tyrosinase activity of the hydrolysates obtained with malate under the same conditions were about 99.5% and 99.3%, respectively. The APTT as anticoagulant activity of carrageenan hydrolysates with 0.3% malate and citrate at $80^{\circ}C$ for 180 min were $2,451{\pm}18(sec)\;and\;1,617{\pm}15(sec)$, respectively.

• Journal of Life Science
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• v.25 no.4
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• pp.425-432
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• 2015

This study was performed to develop high-value-added biomaterials for health and beauty products. Extracts of ethanol and hot water and their subsequent organic solvent fractions were prepared from Lees of Wookukseng (LW), a commercialized Korean traditional rice wine. We investigated their activities on blood coagulation, platelet aggregation, hemolysis against human red blood cells (hRBCs), and anti-oxidation. The water content, pH and brix of the LW were 80.3%, 3.94 and 13.0°, respectively. The yield of ethanol extraction (6.62%) was 3.15 times higher than that of hot-water extraction (2.1%), and the ethyacetate fraction (EAF) of ethanol extract showed the highest content of total polyphenol (128 mg/g) among the various fractions. In anticoagulation activity assay, the EAF of ethanol extract showed a 15-fold extension in TT, PT, and aPTT, indicating that the EAFs contain various inhibitory substances against thrombin, prothrombin and coagulation factors. In anti-platelet aggregation activity assay, the butanol fraction and water residue of ethanol extract showed significant inhibition activity. The activities were comparable to aspirin, a commercial anti-thrombosis agent. The above extracts and fractions did not show hemolysis activity against hRBC up to 5 mg/ml, and had radical scavenging activity against DPPH anion, ABTS cation and nitrite. Our results suggest that the active fractions prepared from LW, which has no specific usage until now, have a high potential as novel resources for anti-thrombosis agents.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.336-343
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• 1999

Fifty six mycelial cultured Basidiomycetes were screened for their inhibitory effects against prolyl endopeptidase(PEP), acetylcholine esterase(AChE) and thrombus coagulation. Out of them, methanolic extract of mycelium and/or ethylacetate(EtOAc) soluble fraction from culture broth of Peniophora quercina, Amanita aspera, Phellinus chrysoloma, Grifola frondosa, Wolfiporia extensa, Clavicorona pyxidata and Phanerochaete sordida inhibited more than 90% of PEP activity at 40 ppm. The extracts of Lenzites betulina, Phellinus chrysoloma, Wolfiporia extensa, Phanerochaete sorrlida, Hypocrea nigricans, Coriolus azureus, Flammulina velutipes, Phlebiopsis gigantea and Bondarzewia montana exhibited about 40% of inhibitory activity against AChE at 40 ppm. In thrombin times assay, the extracts of Amanita aspera, Oxyporus latemarginata, Peniophora quercina, Fomes fomenfarius, Trametes versicolor, and Phlebiopsis gigantea delayed coagulation of thrombus about two to three times over control at ca 550 ppm. In activated partial thromboplastin times assay, none of the tested Basidiomycetes showed significant effect.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.433-438
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• 2006

A fibrinolytic enzyme gene (BCF-1) was subcloned to the pEB vector which is high expression vector in the Bacillus host. The enzyme was purified by using FPLC after ammonium sulfate precipitation. The enzyme was oral-administrated to the rat and checked the bleeding time, blood clotting time and fibrinolytic effect of the serum. In the bleeding time retardation test, it was longer about 1.7 fold in the feeding rat than without feeding. The serum of rat feeded with the enzyme had the fibrinolytic activity from 1 hour to 3 hours after oral-administration. After 3 hours from feeding, the fibrinolytic activity was decreased gradually. Also blood clotting time after bleeding was longer than that of control rat. The enzyme could be detected at band of 30,000 Da in the blood by western blotting. The enzyme was not harmful to the all internal organs of the rats. Taken together, the enzyme originated from B. subtilis BB-1 can be a candidate to develop the drug for thrombosis, arteriosclerosis and myocardial infarction.

• Journal of Life Science
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• v.21 no.10
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• pp.1422-1427
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• 2011

This study was performed to develop effective antithrombotic agents from traditional herb extracts. Prunella vulgaris L. has been used traditionally as a medical resource in cancer therapy, as well as treatment of hypertension and inflammation, and as a diuretic. However, the effects of Prunella vulgaris on thrombosis and platelet activation have not been clearly understood. Antithrombotic and antiplatelet activities of oriental medicinal herbs were investigated by evaluating the effect of the aqueous extract from Prunella vulgaris on the blood coagulation, platelet aggregation and fibrinolysis. Prunella vulgaris extracts showed effective anticoagulant activity in coagulation times such as activated partial thromboplastin time (APTT) and prothrombin time (PT). Prunella vulgaris also inhibited adenosine diphosphate (ADP)- and collagen-induced platelet aggregation. In addition, evaluation of fibrinolytic activity showed that the Prunella vulgaris extracts have high solubility. From these results, it is suggested that Prunella vulgaris can be a potential candidate for anticoagulants and antiplatelets, as well as fibrinolytic agents.

• Journal of Chest Surgery
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• v.29 no.3
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• pp.303-310
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• 1996

From December of 1994 to April of 1995, we, SHUH Department of Pediatric Thoracic and Cardiovascular Surgery, studied effects of aprotinin. 95 patients were randomly divided into two groups : group I (n=47) with aprotinin and group ll (n=48) without aprotinin. Aprotinin was given as one shot injection to cardiopulmonary bypass perfusion solution with dose of 50,000 KIUikg. Laboratory data such as hemoglobin, hematocrit, BUH, creatinine, fibrinogen, electrolyte concentration, aPTT, PT, and AT R was checked preoperatively, 5 minutes after anesthesia, 5 minutes and 35 minutes after CPB circulation, and 5 minutes, 3 hours, and 24 hours after reperfusion. Also, chest-tube drainage, transfused amount of RBC, platelet concentrate, and fresh frozen plasma within first 24 hours postoperatively were checked and analyzed after transition nn body weight demension. Only RBC transfused postoperatively had statistical significance with P value of less than 0.001. Others had no difference statistical wise. Postoperative side effects of aprotinin was not detected weeks after the surgery and there was no reoperated patient due to postoperative bleeding.