• Title/Summary/Keyword: Anticoagulant protein

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Plant Molecular Farming Using Oleosin Partitioning Technology in Oilseeds

  • Moloney, Maurice-M.
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.4
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    • pp.197-201
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    • 1997
  • Plant seed oil-bodies or oleosomes ate the repository of the neutral lipid stored in seeds. These organelles in many oilseeds may comprise half of the total cellular volume. Oleosomes are surrounded by a half-unit membrane of phospholipid into which are embedded proteins called oleosins. Oleosins are present at high density on the oil-body surface and after storage proteins comprise the most abundant proteins in oilseeds. Oleosins are specifically targeted and anchored to oil-bodies after co-translation on the ER. It has been shown that the amino-acid sequences responsible for this unique targeting reside primarily in the central hydrophobic tore of the oleosin polypeptide. In addition, a signal-like sequence is found near the junction of the hydrophobic domain and ann N-terminal hydrophilic / amphipathic domain. This "signal" which is uncleaved is also essential for correct targeting. Oil-bodies and their associated oleosins may be recovered by floatation centrifugation of aqueous seed extracts. This simple partitioning step results in a dramatic enrichment for oleosins in the oil-body fraction. In the light of these properties, we reasoned that it would be feasible to create fusion proteins on oil-bodies comprising oleosins and an additional valuable protein of pharmaceutical or industrial interest. It was further postulated that if these proteins were displayed on the outer surface of oil-bodies, it would be possible to release them from the purified oil-bodies using chemical or proteolytic cleavage. This could result in a simple means of recovering high-value protein from seeds at a significant (i.e. commercial) scale. This procedure has been successfully reduced to practice for a wide variety of proteins of therapeutic, industrial and food no. The utillity of the method will be discussed using a blood anticoagulant, hirudin, and industrial enzymes as key examples.

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Chemical Properties and Physiological Aacitivities of Stromata of Cordyceps militaris

  • Yu, Kwang-Won;Suh, Hyung-Joo;Bae, Song-Hwan;Lee, Chul-Soo;Kim, Sung-Hyun;Yoon, Cheol-Sik
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.266-274
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    • 2001
  • The chemical properties and physiological activities of the freeze-dried stromata of Cordyceps militaris were examined. A proximate analysis exhibited that the stromata consisted mainly of crude carbohydrate (74.3%), crude protein (11.5%), and moisture content (8.7%), with a low content of crude ash (4.%) and fat (1.3%). The carbohydrate was mostly composed of glucose (88.6%). A large quantity of essential fatty acids, including linolenic acid (33.8%) was also observed. An analysis of the component amino acid showed a relatively high ratio of the essential amino acids, lysine (101.2 mg/g), methionine (62.7 mg/g), and acidic amino acids of glutamic aicd (57.5 mg/g) and aspartic acid (43.9 mg/g), whereas a low of tyrosine content (4.7 mg/g). An examination of the cordycepin content indicated tht the stromata and mycelium-embedded media (silkworm pupae) contained 0.2% and 0.5%, respectively. From the examination of the physiological activities, based on methanol extract (M), ethylacetate extract (EA), and hot-water extract (HW) fractions of the stromata, the hot-water extract (HW) fraction showed the most potent intestinal immune modulating activity, anticoagulant activity, and anticomplementary activity, whereas the ethylacetate extract (EA) fraction exhihited a radical scavenging activity. Therefore, the results from the present study indicate that the stromata of C. militaris contain various healthy chemical ingredients, and that especially boiled water extract of whole body would apprear to provide beneficial physiological activities.

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Chemical Properties and Physiological Activities of Synnemata of Beauveria bassiana

  • Yoon, Cheol-Sik;Yu, Kwang-Won;Bae, Song-Hwan;Song, Hyuk-Hwan;Park, Hyun-Soo;Lee, Chan
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.125-133
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    • 2003
  • Chemical properties and physiological activities of the freeze-dried synnemata of Beauveria bassiana were examined. A proximate analysis showed that the synnemata consisted mainly of carbohydrate (49.86%), protein 11.36%), and a moisture content of 30.64%. It contained a low amount of crude ash (4.76%) and fat (3.38%). The carbohydrate was composed mainly of mannose (52.3%), galactose (31.5%), glucose (13.2%), and rhamnose (3%). Trace amounts of arabinose, xylose, and fructose were present. Major amino acids In the synnemata were glutamic acid, glycine, aspartic acid, arginine, threonine, alanine, valine, leucine, lysine, and aspartic acid with the amounts of 30.42, 25.22, 17.17, 15.12, 12.65, 15.23, 12.47, 11.47, 14.24, and 17.17 mg/g, respectively. Among extracts from the synnemata, the hot-water extract showed 67% of anticomplementary activity compared to that of the positive control, followed by ethyl acetate extract (17%) and methanol extract (15%). The hot-water extract also had anticoagulant activity with 55 sec of coagulating time and this fraction exhibited the most potent Intestinal immune system modulating activity. The methanol extract showed the highest inhibitory activity (25%) on the 12-O-tetradecanoyl phorbol-13-acetate-induced superoxide ($O_2^-$) generation, followed by hot-water extract (18%) and ethyl acetate extract (10%). The data in the present study indicate that the extract of Beauveria bassiana synnemata contains some healthful chemical ingredients and it could provide beneficial physiological activities. These features of the synnemata should be of interest to the food industry as well as other industrial fields.

Physiological Properties of Extracts and the Chemical Composition of Tetraselmis sp. JK-46 Cultured with Deep Seawater (해양심층수로 배양한 해양미세조류 Tetraselmis sp. JK-46의 성분 조성 및 생리활성)

  • Joo, Dong-Sik;Kim, Kwang-Woo;Cho, Soon-Yeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.1
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    • pp.1-7
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    • 2011
  • This study examined Tetraselmis sp. JK-46 isolated from seawater from the East Sea. Deep seawater (DSW) had a greater effect on the growth of Tetraselmis sp. JK-46 than surface seawater (SSW). The crude protein, lipid, carbohydrate and ash contents of Tetraselmis sp. JK-46 cultured with DSW were 27.2, 37.1, 13.2 and 26.3 %, respectively, and these values were similar to the results for samples cultured with SSW. The contents of Mg, Ca, Fe and K in the DSW cultured samples were 7080.3, 1009.6, 251.2, and 2749.7 mg/100 g, respectively. The fatty acid compositions of Tetraselmis sp. JK-46 cultured with DSW and SSW were 53.7 and 49.0 % polyunsaturated fatty acids (PUFA) and 25.7 and 30.7 % saturated fatty acids (SFA), respectively. The total amino acid contents of the samples cultured with DSW and SSW were 7392.6 and 6376.0 mg/100 g respectively. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of Tetraselmis sp. JK-46 extracts increased with the concentration of the chloroform and ethyl acetate fractions. The half maximal inhibitiory concentrations ($IC_{50}$) of the chloroform and ethyl acetate fractions of DSW and SSW cultured samples were 1.2 and 2.6 mg/mL, and 3.1 and 3.3 mg/mL, respectively. The ethyl acetate fractions of DSW and SSW cultured samples has anticoagulant activity and the activated partial thromboplastin times (APTT) were 93.4 and 89.3 sec., respectively. The chloroform and ethyl acetate fractions showed antimicrobial activity against Bacillus subtilis, Escherichia coli and Candida albicans.

Antithrombin Active Polysaccharide Isolated from the Alkaline Extract of Red Ginseng

  • Kim Dong Chung;In Man-Jin;Lee Ji Young;Hwang Yoon Kyung;Lee Sung Dong
    • Journal of Ginseng Research
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    • v.23 no.4
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    • pp.217-221
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    • 1999
  • We have isolated an antithrombin active polysaccharide in red ginseng by procedures comprising three major steps involving alkaline extraction, anion exchange and gel permeation chromatography. Active polysaccharide behaved as a single band on cellulose acetate membrane electrophoresis. The average molecular mass was estimated to be about 177 kDa by gel filtration. This polysaccharide was found to be an acidic heteropolysaccharide that contains uronic acid moiety $(40.2\%)$, sulfate group $(9.2\%)$ and protein $(1.5\%)$ in addition to neutral sugar consisted of rhamnose, mannose, galactose, arabinose, glucose, fucose and xylose in a molar ratio of 1.00 : 0.88 : 0.86 : 0.78: 0.70 : 0.33 : 0.22. This polysaccharide inhibited blood coagulation via the intrinsic pathway like heparin in a dose-dependent manner. The clotting of fibrinogen by thrombin was also mitigated by the presence of this polysaccharide.

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Antiarrhythmic effects of ginsenoside Rg2 on calcium chloride-induced arrhythmias without oral toxicity

  • Gou, Dongxia;Pei, Xuejing;Wang, Jiao;Wang, Yue;Hu, Chenxing;Song, Chengcheng;Cui, Sisi;Zhou, Yifa
    • Journal of Ginseng Research
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    • v.44 no.5
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    • pp.717-724
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    • 2020
  • Background: Malignant arrhythmias require drug therapy. However, most of the currently available antiarrhythmic drugs have significant side effects. Ginsenoside Rg2 exhibits excellent cardioprotective effects and appears to be a promising candidate for cardiovascular drug development. So far, the oral toxicity and antiarrhythmic effects of Rg2 have not been evaluated. Methods: Acute oral toxicity of Rg2 was assessed by the Limit Test method in mice. Subchronic oral toxicity was determined by repeated dose 28-day toxicity study in rats. Antiarrhythmic activities of Rg2 were evaluated in calcium chloride-induced arrhythmic rats. Antiarrhythmic mechanism of Rg2 was investigated in arrhythmic rats and H9c2 cardiomyocytes. Results: The results of toxicity studies indicated that Rg2 exhibited no single-dose (10 g/kg) acute oral toxicity. And 28-day repeated dose treatment with Rg2 (1.75, 3.5 and 5 g/kg/d) demonstrated minimal, if any, subchronic toxicity. Serum biochemical examination showed that total cholesterol in the high-dose cohort was dramatically decreased, whereas prothrombin time was increased at Day 28, suggesting that Rg2 might regulate lipid metabolism and have a potential anticoagulant effect. Moreover, pretreatment with Rg2 showed antiarrhythmic effects on the rat model of calcium chloride induced arrhythmia, in terms of the reduced duration time, mortality, and incidence of malignant arrhythmias. The antiarrhythmic mechanism of Rg2 might be the inhibition of calcium influx through L-type calcium channels by suppressing the phosphorylation of Ca2+/calmodulin-dependent protein kinase II. Conclusion: Our findings support the development of Rg2 as a promising antiarrhythmic drug with fewer side effects for clinical use.

Warfarin-induced Skin Necrosis After Valve Surgery (판막수술 후 항응고제 투여로 인한 피부괴사증)

  • Moon, Seung-Chul;Lee, Gun;Lee, Hyeon-Jae;Ahn, Dae-Ho;Lim, Chang-Young
    • Journal of Chest Surgery
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    • v.32 no.3
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    • pp.307-309
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    • 1999
  • Warfarin-induced skin necrosis is a rare complication caused by transient hypercoagulable state. This state is a result of rapid decline of the protein C activity relative to that of coagulation factor II, IX, and X during initiation of oral anticoagulant therapy. We experienced a case of warfarin-induced skin necrosis involving both breasts in a patient who underwent double valve replacement 1 month before. Warfarin was replaced to a low- molecular weight heparin and the necrotic breast lesion was healed spontaneously. Low-dose warfarin was restarted and gradually increased, after which a low molecular weight heparin discontinued..

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Anti-inflammatory effect of ethanol extract from Ecklomia cava on gram-positive bacteria(Peptidoglycan)-induced macrophages (그람-양성균(Peptidoglycan)으로 유도한 대식세포에서 감태 에탄올 추출물의 항염증 효과)

  • Kang, Ok-Hwa;Kim, Sung-Bae;Keum, Joon-Ho;Mun, Su-Hyun;Kim, Yong-Sik;An, Byung-Kwan;An, Hyeon-Jin;Kwon, Dong-Yeul
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.195-205
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    • 2011
  • Objectives : Ecklonia cava is brown alga(Laminariaceae) which grows is sea, it has antioxidant, diarrhea and anticoagulant effect. In this study, the effect of ethanol extract of Ecklonia cava (EC) on peptidoglycan(PGN) -induced NO production in RAW 264.7 cells was investigated. Methods : In the present study, IL-6 production was measured using the enzyme-linked immunosorbent assay(ELISA), prostaglandin $\E_2$($\PGE_2$) production was measured using the EIA kit, and inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) activation, as determined by western blot analysis and reverse transcription -polymerase chain reaction(RT-PCR). Results : EC inhibited PGN-induced NO and IL-6 production. Consistent with these observations, the protein expression of iNOS and COX-2 were inhibited by EC. Moreover, EC suppressed the phosphorylation of extracellular signal-regulated kinase(ERK) 1/2 in PGN-induced RAW 264.7. Conclusions : These results suggest that EC has inhibitory effects on PGN-induced $\PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

Nafamostat Mesilate Inhibits TNF-${\alpha}$-Induced Vascular Endothelial Cell Dysfunction by Inhibiting Reactive Oxygen Species Production

  • Kang, Min-Woong;Song, Hee-Jung;Kang, Shin Kwang;Kim, Yonghwan;Jung, Saet-Byel;Jee, Sungju;Moon, Jae Young;Suh, Kwang-Sun;Lee, Sang Do;Jeon, Byeong Hwa;Kim, Cuk-Seong
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.3
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    • pp.229-234
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    • 2015
  • Nafamostat mesilate (NM) is a serine protease inhibitor with anticoagulant and anti-inflammatory effects. NM has been used in Asia for anticoagulation during extracorporeal circulation in patients undergoing continuous renal replacement therapy and extra corporeal membrane oxygenation. Oxidative stress is an independent risk factor for atherosclerotic vascular disease and is associated with vascular endothelial function. We investigated whether NM could inhibit endothelial dysfunction induced by tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$ ). Human umbilical vein endothelial cells (HUVECs) were treated with TNF-${\alpha}$ for 24 h. The effects of NM on monocyte adhesion, vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) protein expression, p38 mitogenactivated protein kinase (MAPK) activation, and intracellular superoxide production were then examined. NM ($0.01{\sim}100{\mu}g/mL$) did not affect HUVEC viability; however, it inhibited the increases in reactive oxygen species (ROS) production and p66shc expression elicited by TNF-${\alpha}$ (3 ng/mL), and it dose dependently prevented the TNF-${\alpha}$ -induced upregulation of endothelial VCAM-1 and ICAM-1. In addition, it mitigated TNF-${\alpha}$ -induced p38 MAPK phosphorylation and the adhesion of U937 monocytes. These data suggest that NM mitigates TNF-${\alpha}$ -induced monocyte adhesion and the expression of endothelial cell adhesion molecules, and that the anti-adhesive effect of NM is mediated through the inhibition of p66shc, ROS production, and p38 MAPK activation.

The Clinical Significance of Antiphospholipid Antibodies in Korean Children with Henoch-$Sch{\ddot{o}}nlein$ Purpura (Henoch-$Sch{\ddot{o}}nlein$ 자반증에서 항인지질 항체의 임상적 의의)

  • Park, Eun-Jung;Baek, Ji-Young;Shin, Jae-Il;Lee, Jae-Seung;Kim, Hyon-Suk
    • Childhood Kidney Diseases
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    • v.13 no.2
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    • pp.146-152
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    • 2009
  • Purpose : We performed this study to evaluate the incidence and clinical significance of antiphospholipid antibodies (aPL Ab) in Korean children with Henoch-$Sch{\ddot{o}}nlein$ purpura (HSP). Methods : The medical records of 62 patients (31 boys and 31 girls) aged $46.0{\pm}3.1$ (1-16) years with a clinical diagnosis of HSP based on the EULAR/PReS criteria were reviewed retrospectively. From the years 2007 to 2009, the sera from children with acute HSP were tested for aPL Ab such as LA, anti-cardiolipin antibody and anti-${\beta}_2$ glycoprotein I antibody. Results : LA was positive in 18 (29%) of the 62 patients with HSP and We divided the patients into the two groups LA positive group (N=18) and LA negative group (N=44). There were no significant differences between the two groups with regard to abdominal pain, arthralgia and renal involvement, but LA positive group had significantly higher C-reactive protein ($4.3{\pm}7.2$ mg/dL vs. $1.3{\pm}1.8$ mg/dL, P=0.035), erythrocyte sedimentation rate ($37.5{\pm}26.2$ mm/hr vs. $25.1{\pm}22.6$ mm/hr, P= 0.039), IgM ($148.1{\pm}48.4$ mg/dL vs. $114.9{\pm}41.5$ mg/dL, P=0.024), C3 ($143.1{\pm}21.9$ mg/dL vs. $129.7{\pm}24.5$ mg/dL, P=0.048) and C4 levels ($30.9{\pm}6.3$ mg/dL vs. $24.9{\pm}7.8$ mg/dL, P=0.002) compared with LA negative group. Conclusion : We found that the incidence of positive aPL Ab tests was relatively higher in Korean children with HSP and the presence of aPL Ab was associated with acute inflammatory process of HSP. These results suggest that the aPL Ab are involved in the pathogenesis of HSP in children.