• Tuberculosis and Respiratory Diseases
• /
• v.40 no.2
• /
• pp.123-134
• /
• 1993

Background: Tissue-type plasminogen activator is a physiologic activator, which has high affinity for fibrin and is activated by fibrin. Because of these properties, t-PA has the potential to induce effective thrombolysis without producing a systemic lytic state. In practice, however, therapeutically efficacious doses of t-PA has been associated with the development of a systemic lytic state. As experience with t-PA has accumulated, it has suggested that the fibrin selectivity is influenced by the dose and duration of t-PA infusion, and many studies have performed in an attempt to optimize the duration of t-PA regimen. Methods: This study was designed to assess the thrombolytic efficacy of t-PA and the differences of two dosing regimens of t-PA (infusion of 1 mg/kg t-PA over 15 or 180 minutes) in a canine model of pulmonary embolism, induced by injection of radioactive autologous blood clots. By continuously counting over both lung fields with a external gamma counter, we correlated rate and extent of pulmonary thrombolysis with corresponding pulmonary hemodynamics in addition to the gas analyses of arterial and mixed venous blood. Results: 1) While total clot lysis was similar ($36.2{\pm}3.3%$ and $39.6{\pm}2.3%$ respectively, p>0.05) when t-PA was infused over 15 or 180 minutes, the rate of lysis during infusion was markedly increased with the shorter infusion ($81.4{\pm}16.8%/hr$ vs $37.3{\pm}2.4%/hr$, p<0.05). 2) The duration of thrombolysis was $63.3{\pm}22.2$ minutes although t-PA was administered over 15 minutes, and it was only $148.5{\pm}14.0$ minutes in case of the infusion over 180 minutes (p<0.05). 3) The increased rate of thrombolysis with the shorter infusion was accompanied by a faster amelioration of cardiopulmonary impairment from pulmonary embolism (p<0.05). Conclusion: It is concluded that the shorter (15 minutes) infusion of t-PA is superior to the longer (180 minutes) infusion when the dose is equal, in consideration of the faster improvement in cardiopulmonary impairment from pulmonary embolism.

• Journal of Integrative Natural Science
• /
• v.7 no.3
• /
• pp.173-182
• /
• 2014

To simplify the different biological investigation of the methanolic extract and solvent-solvent partitioning of Lablab purpures (L. purpures) bark. In-vitro anti-oxidant study was determined using total DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing zone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and thrombolytic activity by clot disruption method. The antioxidant potential was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteau reagents using butylated hydroxytolune (BHT) and ascorbic acid as standards. The Aqueous soluble fraction revealed the highest free radical scavenging activity ($IC_{50}=48.76{\mu}g/mL$). The antimicrobial screening of the bark of L. purpures exhibited mild to moderate activity in test microorganisms. The CSF showed the maximum relative percentage inhibition against Salmonella parathyphi (34.2%) for bacteria and C. albicans (28.8%) for fungi whereas, lowest relative percentage inhibition against Sarcina lutea (22.0%) for bacteria and Aspergillus niger (24.4%) for fungi. In the brine shrimp lethality bioassay, The $LC_{50}$ values of Carbon tetrachloride and N-Hexane soluble fraction were found $92.18{\mu}g/mL$, and $68.95{\mu}g/mL$ respectively while the $LC_{50}$ values of standard Vincristine sulphate was $1.37{\mu}g/mL$. The methanolic extract and its organic soluble fractions of Lablab purpureus at concentration 2.0 mg/mL, significantly protected the lysis of erythrocyte membrane induced by hypotonic solution and heat as compared to the standard, acetyl salicylic acid (0.10 mg/mL). The MSF and AQSF produced 61.48 % and 53.75% inhibition of hemolysis of RBC caused by hypotonic solution respectively, whereas acetyl salicylic acid (0.10 mg/mL) showed 76.42%. Ethanol extract of L. purpures and all of its different partitions exhibited moderate thrombolytic activity of 37.25%-2.40%. Very good preliminary screening and simplified experiments were able to show the different biological activity of methanolic extract and its soluble fractions of L. purpures at a time.

• Korean Journal of Food Science and Technology
• /
• v.44 no.5
• /
• pp.600-606
• /
• 2012

A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

• Journal of Life Science
• /
• v.28 no.9
• /
• pp.1068-1075
• /
• 2018

Kaempferia parviflora, an herbaceous plant in the family Zingiberaceae, is popular in many tropical regions. It is called as black ginger or krachaidum in Thailand and Laos, and its raw or dried root have been used as spices and teas. The rhizomes also have been traditionally used to treat gastrointestinal disorders, ulcers, gout, dysentery, allergies and to improve physical work capacity. Recently, its anti-obesity, anti-oxidant, anti-inflammatory and blood clot-lysis activities were reported. In this study, the anti-thrombosis activity of black ginger was investigated, since improvement in blood fluidity leads to the prevention of various lifestyle-related diseases. The hot water and ethanol extract and their subsequent solvent fractions (hexane, ethylacetate, butanol fractions and water residue) were prepared, and their anti-coagulation and platelet aggregation inhibitory activities were determined, respectively. Among the black ginger extracts and their fractions, the ethylacetate fraction (EAF) of ethanol extract only showed significant extensions of blood coagulation time determined by thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (aPTT). At 5 mg/ml concentration, TT, PT and aPTT were extended to 1.22, 1.49 and >15-folds compared to non-treatment. The EAFs of ethanol and hot water extract showed strong inhibitions against collagen-induced platelet aggregations, which are comparable to inhibitions of aspirin. Also the EAFs from black ginger did not show any hemolysis activity against human RBC up to 0.5 mg/ml. Our results suggest that the EAF of black ginger has a potential as novel anti-coagulation and ant-platelet aggregation agent. This report provides the first evidence of anti-coagulation activity of black ginger.

• Journal of Plant Biotechnology
• /
• v.36 no.1
• /
• pp.30-37
• /
• 2009

Tissue-type plasminogen activator (t-PA) is a thrombolytic agent important in fibirn clot lysis. T-PA causes fibirn-specific plasminogen activation. Six binary vectors harboring t-PA and its derivative genes were cloned and expressed in transgenic alfalfa plants. The insertion of the t-PA and its derivative genes in genomic DNA of alfalfa plants was confirmed by PCR. The presence of the t-PA and its derivative transcripts in total RNAs of the transgenic alfalfa leaves was verified by RT-PCR. ELISA experiments demonstrated that the highest level of recombinant t-PA expression was $75.1{\mu}g$/ total soluble protein (mg) in alfalfa plants. The amount of recombinant t-PA and its derivative proteins in transgenic plants was estimated to range from 9.7 to $39.5{\mu}g$/ total soluble proteins (mg). Western blot analysis of the transformed alfalfa leaves revealed bands of approximately 68-kDa recombinant t-PA and its derivative proteins. The fibrinolysis of recombinant t-PA and its derivative proteins was confirmed by a fibrin plate assay (range from 3.2 to 8.1 cm). The results presented provide information for the development of an additional production of recombinant human proteins having pharmaceutical applications using transgenic plants.