• Archives of Pharmacal Research
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• v.21 no.4
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• pp.374-377
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• 1998

The freeze-dried powder of Lumbricus rubellus earthworm was administered orally to rats and its fibrinolytic and antithrombotic effects were investigated. The fibrinolytic activity of plasma was determined by measuring the plasmin activity of the euglobulin fraction and was increased to two-folds of the control at a dose of 0.5 g/kg/day and five times with 1 g/kg/day after 4-day administration. The antithrombotic effect was studied in an arterio-venous shunt model of rats. The thrombus weight decreased significantly from 43.2 mg to 32.4 mg at a dose of 0.5 g/kg/day after 8-day treatment. The level of fibrinogen/fibrin degradation product (FDP) in serum was elevated in a dose-dependent manner during the treatment period. On the 8th day after administration, the FDP value was increased to 7.7 $\mu\textrm{g}$g/ml compared with the control value of 3.3 $\mu\textrm{g}$g/ml. These results support that earthworm powder is valuable for the prevention and/or treatment of thrombotic conditions.

• KSBB Journal
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• v.27 no.6
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• pp.375-380
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• 2012

We previously reported that Ultra nattokinase$^{(R)}$ showed high fibrinolytic activity and revealed antithrombotic effect in rat blood plasma based on its ability to suppress collagen-induced platelet aggregation. This research was carried out to verify the clot lysing activity and blood flow enhancing effects of Ultra nattokinase$^{(R)}$ via monitoring and comparing the antithrombotic effects in rat artery between oral administration of Ultra nattokinase$^{(R)}$ and maltodextrin. SD rats were fed with 1.11 mg/kg of Ultra nattokinase$^{(R)}$ for 4 weeks. The effect on arterial thrombosis was then evaluated using an antithrombotic model after induction by $FeCl_3$. Detected fibrinolytic activity was proportional to the content of Ultra nattokinase$^{(R)}$ and statistical extents of the antithrombotic activity was enhanced strongly twice rather than control group. The PT and the aPTT, however, showed only a small difference between two groups. The results suggest that Ultra nattokinase$^{(R)}$ can effectively treat thromboembolism and enhance blood flow, and that Ultra nattokinase$^{(R)}$ can also prevent venous occlusion by aiding clot lysis.

• Journal of Mushroom
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• v.13 no.1
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• pp.30-36
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• 2015

Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate anti-oxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.327-337
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• 2021

Fibrinolytic enzymes with a direct mechanism of action and safer properties are currently requested for thrombolytic therapy. This paper reports on a new enzyme capable of degrading blood clots directly without impairing blood coagulation. This enzyme is also non-cytotoxic and constitutes an alternative to other thrombolytic enzymes known to cause undesired side effects. Twenty-four Bacillus isolates were screened for production of fibrinolytic enzymes using a fibrin agar plate. Based on produced activity, isolate S127e was selected and identified as B. subtilis using the 16S rDNA gene sequence. This strain is of biotechnological interest for producing high fibrinolytic yield and consequently has potential in the industrial field. The purified fibrinolytic enzyme has a molecular mass of 27.3 kDa, a predicted pI of 6.6, and a maximal affinity for Ala-Ala-Pro-Phe. This enzyme was almost completely inhibited by chymostatin with optimal activity at 48℃ and pH 7. Specific subtilisin features were found in the gene sequence, indicating that this enzyme belongs to the BPN group of the S8 subtilisin family and was assigned as AprE127. This subtilisin increased thromboplastin time by 3.7% (37.6 to 39 s) and prothrombin time by 3.2% (12.6 to 13 s), both within normal ranges. In a whole blood euglobulin assay, this enzyme did not impair coagulation but reduced lysis time significantly. Moreover, in an in vitro assay, AprE127 completely dissolved a thrombus of about 1 cc within 50 min and, in vivo, reduced a thrombus prompted in a rat tail by 11.4% in 24 h compared to non-treated animals.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.274-281
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• 2006

A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

• Journal of Life Science
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• v.25 no.12
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• pp.1432-1438
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• 2015

Although exercise training has been utilized to improve vascular function in animals and humans, the impact of moderate intensity exercise training on fibrinolytic activities and nitric oxide (NO) bioavailability has not been well documented. Therefore, the purpose of the current study was to examine the impact of moderate intensity aerobic exercise training on fat mass, blood lipid profiles, fibrinolytic activity, and NO levels in high-fat-diet induced rats. The body weight, fat mass, blood lipid profiles, fibrinolytic activity, and nitrite/nitrate were measured pre- and postexercise (10 weeks) training. The body weight and fat mass reduced significantly in the exercise (EX) group compared to the control (CON) group. Blood lipid profiles and low-density lipoprotein were unchanged in the EX group compared to the CON group. However, triglyceride and free fatty acid were significantly lower in the EX group compared to the CON group, and high-density lipoprotein was significantly greater in the EX group compared to the CON group. In addition, fibrinolytic activity and nitrite/nitrate were significantly greater in the EX compared to the CON group. These results suggest that 10 weeks of the moderated intensity aerobic exercise training improves blood lipid profiles, fibrinolytic activity, and the nitrite/nitrate ratio, which may improve vascular health and reduce obesity-related cardiovascular disease risks in high-fat- diet induced rats.

• Journal of Biomedical Engineering Research
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• v.26 no.3
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• pp.157-161
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• 2005

Direct injection of a fibrinolytic agent to the intraarterial thrombosis may increase the effectiveness of thrombolysis by enhancing the permeation of thrombolytic agents into the blood clot. Permeation of fibrinolytic agents into a clot is influenced by the surface pressure, which is determined by the injection velocity of fibrinolytic agents. In order to calculate the pressure distribution on the clot surface for different jet velocities (1, 3, 5 m/sec) and nozzle arrangements (1, 9, 17 nozzles), computational fluid dynamic methods were used. Thrombolysis of a clot was mathematically modeled based on the pressure and lysis front velocity relationship. Direct injection of a thrombolytic agent increased the speed of thrombolysis significantly and the effectiveness was increased as the ejecting velocity increased. The nine nozzles model showed about $20\%$ increase of the lysed volume, and the one and seventeen nozzles models did not show significant differences. The wall shear stress decreased as the number of nozzles increased, and the wall shear stress in most vessel wall was lower than 25 Pa. The results implied that thrombolysis could be accelerated by direct injection of a drug with the moderate velocity without damaging the blood vessel wall.

• Tuberculosis and Respiratory Diseases
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• v.76 no.3
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• pp.127-130
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• 2014

The risk of dying from a pulmonary embolism (PE) is estimated to be about 30% if inotropic support is required and no cardiopulmonary arrest occurs. Fibrinolysis in massive PE is regarded as a life-saving intervention, unless there is a high risk of bleeding following the use of the fibrinolytic therapy. Rivaroxaban is an oral factor Xa inhibitor, however its anticoagulation effects before or after administration of fibrinolytics in massive PE are still unknown. Two patents were admitted: 61-year-old woman with repeated syncope, and a 73-year-old woman was admitted with dyspnea and poor oral intake. Systemic arterial hypotension with radiologic confirmation led to a diagnosis of massive PE in both patients. Rivaroxaban was administered before in one, and after firbrinolytic therapy in the other. One showed similar efficacy of rivaroxaban with currently used anticoagulants after successful fibrinolysis, and the other one without antecedent administration of the fibrinolytic agent showed unfavorable efficacy of rivaroxaban.

• Korean Journal of Agricultural Science
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• v.36 no.1
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• pp.111-122
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• 2009

When the cattail pollen was identified by using fibrinolytic agents, we found that the fibrinolytic activity was controlled by an enzyme. Therefore, for determining the fibrinolytic activity of cattail pollen, the fibrinolytic enzyme in cattail pollen was purified by gel filtration using DEAE-cellulose, Sephadex G-150 and HPLC. Also, its purity was certified by polyacrylamide gel electrophoresis, and its physico-chemical properties, such as pH and temperature stabilities and effects of metal, inhibitors and substrates, were examined. The specific activity, purification fold, and molecular weight of the enzyme were 38U/mg, 86.4,and 75kDa, respectively. The optimum pH for the purified enzyme was at 4.0 and it was stable at pH 4.0-6.0. The optimum temperature was $55^{\circ}C$ and it was stable at $30-60^{\circ}C$. But the enzyme began to be inactivated at $70^{\circ}C$ and its activity was totally lost at temperatures above $80^{\circ}C$. As for substrate specificity, the enzyme was most effective in dissolving fibrin, followed by whole casein, ${\kappa}$-casein, ${\alpha}$-casein, ${\beta}$-casein, and BSA. With casein as the substrate, Km value was found to be 0.44mM and the enzyme showed a high affinity for casein. As for the metal ions affecting enzyme activity, $K^+$, $Na^+$, and $Mg^{2+}$ had no effect on enzyme reaction while $Zn^{2+}$ and $Fe^{2+}$ showed potent inhibitory activity. Judging from the fact that the purified enzyme was also strongly inhibited by PMSF, iodoacetic acid, and SDA, it assumed to be a serine protease.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.7
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• pp.140-148
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• 2017

The purpose of this study was to examine the effects of acute aerobic exercise at different intensities on the blood pressure, blood lipids and fibrinolytic markers in pre-hypertension college-aged males. Six subjects performed an acute running exercise at three different intensities(low intensity(LI): 50-60% heart rate reserve(HRR), moderate intensity(MI): 60-70%HRR, and high intensity(HI): 70-80%HRR). The blood pressure(systolic(SBP) and diastolic blood pressure(DBP), blood lipids(total cholesterol(TC) and high-density lipoprotein cholesterol(HDL-C)) and fibrinolytic markers(tissue plasminogen activator(tPA) and plasminogen activator inhibitor-1(PAI-1)) were determined before(PRE), immediately after(POST) and 60minutes after the exercise(60 POST). Results: the SBP in the LI group was significantly increased at POST(p=0.013). The ES levels for the SBP in the MI and HI groups were reduced (-1.33 and -1.23, respectively), though the differences were not significant. The HDL in the MI(p=0.003) and HI(p=0.002) groups were significantly increased at 60 POST. Also, the tPA in the MI(p=0.021) and HI(p=0.042) groups were significantly increased at POST.