• BMB Reports
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• 제30권1호
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• pp.37-40
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• 1997

Earthworm extracts are known for anti-inflammatory, analgesic. antipyretic, and anticancer effects but can also influence blood circulation. It was previously shown that an earthworm, Lumbricus rubelius. contained a water-extractable anticoagulant which was a heat- and acid-stable molecule with hydrophilic property. In order to uncover the biochemical nature of this molecule, the anticoagulant was processed with various hydrolases such as trypsin, DNase, RNase. and lysozome. When the digested samples were analyzed with an in vitro coagulation test measuring activated partial thromboplastin time (APTT) and agarose gel electrophoresis, the anticoagulant proved to be a relatively homogeneous DNA fragment with relative molecular size around 72 base pairs. Interestingly, the activity was further stimulated with a trypsin digestion. RNA. on the other hand, did not prolong the APTT. It was also demonstrated that the DNA accelerated the antithrombin III (AT-III) inhibition of thrombin from $IC_{50}$ of 0.34 to 0.16 unit determined with S-2238 as a substrate, whereas heparin, a popular anticoagulant. shifted the value to 0.05. Therefore, it is suggested that the DNA could be considered as an alternative antithrombotic agent to heparin, which would exhibits bleeding side effects.

• 한국식품영양과학회지
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• 제29권5호
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• pp.908-916
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• 2000

This study was undertaken to purify the anticoagulant polysaccharide from Pachymeniopsis elliptica. The anticoagulant compound (PE-2IVc-2-1) was isolated from the crude polysaccharide (PE-2) by the sequential steps such as ultrafiltration (PE-2IV), anion-exchange chromatography of DEAE-Toyopearl 650C(PE-2IVc), size-exclusion chromatography of Sepharose CL-6B (PE-2IVc), and HPLC Shodex OHpak column chromatography (PE-2IVc-2-1). The apparent molecular weight of purified polysaccharide (PE-2IVc-2-1) was approximately 710 kDa. It was mainly composed of galactose (48.6%) and 3,6-anhydrogalactose (44.0%) with 27.1% of sulfate residue, in addition a trace amount of xylose (3.6%), glucose (1.1%), mannose (1.1%), arabinose (0.8%) and fucose (0.8%). The IR spectrum of the polysaccharide showed the characteristic bands of S=O(1116.74 and 1143.17 $cm^{-1}$) and C-O-S (828.19 $cm^{-1}$) stretching, respectively. The anticoagulant effect of PE-2 in ex vivo was proportional to the concentration of intravenously injected dose, up to 100 mg/kg.

• Applied Biological Chemistry
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• 제42권2호
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• pp.140-146
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• 1999

청각(1.2 kg)을 0.8% 염산 수용액(24 l)으로 추출한 추출물을 메탄올 환류, 에탄올 침전 및 투석하여 활성이 약 2배 증가된 조다당(CF-1)으로부터 2종의 항응고활성 다당류를 정제하였다. 정제는 CF-1의 DEAE-Toyopearl이온교환 크로마토그래피와 Sephadex G-75, Sephadex G-100, Sepharose CL-6B 겔여과 크로마토그래피, HPLC 등을 이용하였다. 최종 정제 다당류획분인 CF-1-VIa-1과 CF-1-VIIa-1는 분자량이 각각 80,000과 40,000 Da 이었으며, 주구성당으로서 arabinose와 galactose가 약 2:1의 몰비율로 풍부하게 함유되어 있었고, 구성당 잔기에 $12{\sim}13%$의 유황을 함유하는 함황성 다당류들이었다. CF-1-VIa-1와 CF-1-VIIa-1의 항응고활성을 $2.5\;{\mu}g/mL$의 농도에서 비교하였을 때 APTT활성은 대조군에 비하여 각각 262초, 250초이었고 TT활성은 각각 130초, 70초이었으며, 분자량이 큰 CF-1-VIa-1이 다소 높은 항응고활성을 보였다. CF-1-VIa-1와 CF-1-VIIa-1의 desulfation과 sulfation을 통한 항응고활성을 비교한 결과 desulfation시 항응고활성이 각각 약 80%와 50%로 감소하였으나 sulfation시에는 약 30%와 20%로 그 활성이 증가하였다. 두 다당류는 헤파린과 달리 농도 의존적으로 불용성 피브린의 형성을 억제하므로써 antithrombin III 비의존적 트롬빈 저해활성을 나타내었으며, 칼슘이온의 킬레이트에 의한 혈액응고 저해효과는 나타내지 않았다.

• 한국식품영양과학회지
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• 제29권2호
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• pp.335-341
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• 2000

허브를 비롯한 식용되는 채소류로부터 항응고 활성을 검색하기 위하여 93종을 대상으로 냉수(Fr. I), 메탄올(Fr. ll), 열수(Fr. lll)추출물을 각각 조제하였다. 내인성 경로에 작용하는 항응고 활성은 수용성 획분인 Fr. I과 Fr.lll를 대상으로 activated partial thromboplastin time(APTT)으로 검색한 결과 양파, 마늘, 정향, 쑥, 울금 등이 높은 활성을 보였으며, 외인성 경로와 공통경로에 작용하는 항응고 활성은 저분자임을 감안하여 Fr. I과 Fr. II를 대상으로 각각 prothrombin time(PT) 과 thrombin time(TT)으로 검색한 결과 민트, 이탈리안 시즈닝, 로즈메리, 타라곤, 울금, 백리향, 와사비는 공통경로에 항응고 활성을 나타내였으나 외인성 경로에 단독으로 항응고 활성을 나타내는 시료는 없었다. 1, 2차 검색 결과 가장 높은 항응고 활성을 보인 정향(Eugenia caryophyllate, clove)을 대상으로 산지를 달리하여 항응고 활성을 검색한 결과 홀랜드산 정향의 활성이 가장 높았다.

• Parasites, Hosts and Diseases
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• 제52권2호
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• pp.183-188
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• 2014

Mosquitoes secrete saliva that contains biological substances, including anticoagulants that counteract a host's hemostatic response and prevent blood clotting during blood feeding. This study aimed to detect heparin, an anticoagulant in Aedes togoi using an immunohistochemical detection method, in the salivary canal, salivary gland, and midgut of male and female mosquitoes. Comparisons showed that female mosquitoes contained higher concentrations of heparin than male mosquitoes. On average, the level of heparin was higher in blood-fed female mosquitoes than in non-blood-fed female mosquitoes. Heparin concentrations were higher in the midgut than in the salivary gland. This indicates presence of heparin in tissues of A. togoi.

• 한국임상수의학회지
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• 제34권3호
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• pp.197-199
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• 2017

A 14-year-old castrated male ShihTzu diagnosed with chronic kidney disease (CKD) 6 months prior was referred to our clinic. The patient had been experiencing symptoms such as vomiting, poor appetite and hind limbs weakness. Hematology tests showed that he had a non-regenerative anemia. With aggressive treatment, the patient's state had gotten worse. He showed ragged breath, vomiting blood and loss of consciousness temporarily. Hematocrit maintained low level. Gastric hemorrhage was strongly suspected by hematemesis. Whole blood transfusion was performed and heparin was used as an anticoagulant. Prior to transfusion, the blood cross matching between donor and patient was performed and the result was compatible. After the transfusion was stabilized, 1 mg of protamine sulfate for each 100 units of heparin was prepared and given intravenously over 3 minutes to reverse the effects of heparin. Immediately after protamine injection, the patient conducted severe anaphylactic shock. Protamine sulfate is used to reverse the anticoagulant action of heparin in dogs and humans. The adverse reaction of protamine sulfate range from mild reaction to fetal cardiac arrest. When using protamine sulfate as heparin neutralization, it can lead to the death of a patient cause of anaphylactic shock. For this reason, the protamine sulfate should be injected slowly with antihistamine and the clinician should carefully monitor patients.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.327.1-327.1
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• 2002

Glycosaminoglycans(PT -Gag) were isolated from the porcine testis. From the PT -Gag, we obtained two different types of Gag fractions using Dowex macro porous Resin MSA-1 column, PT -Gag-1.5% NaCl and PT -Gag-16% NaCl. Various biological activities of the GAGs were examined in aspect of anticoagulant and immunomodulating activity. The anticoagulant activity of the GAGs was evaluated by activated partial thromboplastin time (aPTT ) assay and thrombin time (TT) assay. The GAGs of porcine testis markedly incresed the clotting times of both of aPTT and TT. showing that PT-Gag-16% NaCl was more effective than PT-Gag-1.5% NaCl. The immunomodulating activityof the GAGs was examined in relation to regulation of xytoxine prodution of murine peritoeal maerophages. Taken together. GAGs isolated from porcine testis possess bilolgical functions such as anticoagulant and immunomodulating activity.

• Clinical and Experimental Pediatrics
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• 제57권4호
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• pp.202-205
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• 2014

Lupus anticoagulant-hypoprothrombinemia syndrome (LAHPS), a very rare disease that is caused by the presence of antifactor II antibodies, is usually counterbalanced by the prothrombotic effect of lupus anticoagulant (LAC). Patients with LAHPS are treated using fresh frozen plasma, steroids, immunosuppressive agents, and immunoglobulins for managing the disease and controlling hemorrhages. Notably, steroids are the important treatment for treating hypoprothrombinemia and controlling the bleeding. However, some patients suffer from severe, life-threatening hemorrhages, when factor II levels remain very low in spite of treatment with steroids. Here, we report a case of LAHPS in a 15-year-old girl who experienced pulmonary hemorrhage with rapid progression. She was referred to our hospital owing to easy bruising and prolonged bleeding. She was diagnosed with LAHPS that presented with pancytopenia, positive antinuclear antibody, proloned prothrombin time, activated partial thromboplastin time, positive LAC antibody, and factor II deficiency. Her treatment included massive blood transfusion, high-dose methylprednisolone, vitamin K, and immunoglobulin. However, she died due to uncontrolled pulmonary hemorrhage.

• Archives of Pharmacal Research
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• 제20권1호
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• pp.17-23
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• 1997

A saline suspension of Lumbricus rubellus earthworm powder (EWP) was administered to rats (1 g/kg/day) orally for 15 days to evaluate an oral effectiveness for thrombotic disorders. Blood was drawn at 2-day interval after the administration. Several parameters for antithrombotic, anticoagulant and fibrinolytic activities were measured, including platelet aggregation, clotting time, plasmin activity and the levels of FDP (fibrin/fibrinogen degradation products), D-dimer, and t-PA antigen. It did not affect platelet aggregation induced by ADP and collagen but anticoagulant activity (aPTT and TT) was gradually increased to two-folds for the first 5 days of administration and back to normal. Fibrinolytic activity of euglobulin fraction was highest on the 11 th day after the administration. The level of FDP was elevated to be comparable to the positve control$ (5-10 {\mu}g/ml)$ after 9-day treatment. Oral administration of the EWP could also reduce the formation of venous thrombus induced with viper venom. Complete blood count (CBC) profiles were within normal ranges except for a slight increase in white blood cells after the oral administration for 15 days. These results suggested that the EWP may be valuable for the prevention and/or treatment of thrombotic diseases.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.503-511
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• 2008

An edible marine red alga, Grateloupia filicina, collected from Jeju Island of Korea was hydrolyzed by cheap food-grade carbohydrases (Viscozyme, Celuclast, AMC, Termamyl, and Ultraflo) to investigate its anticoagulant activity. Among the tested enzymatic extracts of G. filicina, a Termamyl extract showed the highest anticoagulant activity. Anion-exchange chromatography on DEAE-cellulose and gel-permeation chromatography on Sepharose-4B were used to purify the active polysaccharide from the crude polysaccharide fraction of G. filicina. The purified sulfated polysaccharide (0.42 sulfate/total sugar) showed ${\sim}1,357kDa$ molecular mass and was comprised mainly of galactose(98%) and 1-2% of glucose. The sample showed potential anticoagulant activity on activated partial thromboplastin time (APTT) thrombin time (TT) assays. The purified G. filicina anticoagulant (GFA) inhibited the coagulation factor X (92%), factor II (82%), and factor VII (68%) of the coagulation cascade, and the molecular interaction (protein-polysaccharide) was highly enhanced in the presence of ATIII (antithrombin III). The dissociation constant of polysaccharide towards serine proteins decreased in the order of FXa (58.9 nM) >FIIa (74.6 nM) >FVII (109.3 nM). The low/less cytotoxicity of the polysaccharide benefits its use in the pharmaceutical industry; however, further studies that would help us to elucidate the mechanism of its activity are needed.