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Purification and Biochemical Characterization of a Novel Fibrinolytic Enzyme from Streptomyces sp. P3

  • Cheng, Guangyan;He, Liying;Sun, Zhibin;Cui, Zhongli;Du, Yingxiang;Kong, Yi
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1449-1459
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    • 2015
  • A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg2+ and Ca2+ and completely inhibited by Cu2+, but slightly enhanced by Fe2+. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.

Antibacterial and Antioxidant activities of Bio-fermented Sanguisorbae Radix Extract (지유 발효추출물의 항세균 및 항산화 활성)

  • Kil, Ki-Jung;Doh, Eun-Soo;Chang, Jun-Pok;Lee, Seung-Hyun;Yoo, Ji-Hyun
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.29-35
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    • 2015
  • Objectives : The objective of this research was to investigate the antibacterial and antioxidant activities of bio-fermented Sanguisorbae Radix extract.Methods : The Sanguisorbae Radix extract was fermented byStreptococcus thermophilus,and their products was tested for antibacterial activity against pathogenic microorganisms namely,Bacillus cereus,Bacillus subtilis,Staphylococcus aureus,Escherichia coli,Vibrio parahaemolyticusandSalmonella typhimuriumby paper disc diffusion method and the antioxidant activities of extract was evaluated by five different assays as electron donating ability(EDA), superoxide dismutase(SOD)-like activity, polyphenol, flavonoid contents and nitrite scavenging ability.Results : The bio-fermented Sanguisorbae Radix extract was safe from heat. Antibacterial activity of fermented Sanguisorbae Radix extract appeared relatively highly againstBacillus cereusandStaphylococcus aureusand didn't show any difference. EDA in comparison to Vitamin C showed over 90% activity at about the same time of Sanguisorbae Radix extract expressed highly. SOD activity showed 15% in fermentation before and after. The nitrite scavenging ability of Sanguisorbae Radix extract before and after fermentation showed higher numerical value over 70% in pH 2.5 than that of butylated hydroxytoluene(BHT). But SOD activity, EDA and nitrite scavenging ability were not different between the Sanguisorbae Radix extract before and after fermentation. Total polyphenol content expressed over about 20 mg/g, and that of the Sanguisorbae Radix extracts was increased than that of the fermented Sanguisorbae Radix extracts.Conclusions : The results suggest the usefulness of developing functional materials using antioxidant active Sanguisorbae Radix extract was fermented bySalmonella typhimuriumwith high polyphenol contents and nitrite scavenging ability.

The Effect of Aspalatone, a New Antithrombotic Agent, on the Specific Activity of Antioxidant Enzyme in the Rat Blood

  • Kim, Chin;Koo, Chang-Hui;Choi, Dong-Young;Cho, Yong-Joon;Choi, Jae-Ho;Im, Doo-Hyeon;Jhoo, Wang-Kee;Kim, Hyoung-Chun
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.348-352
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    • 1996
  • The antioxidant efficacy of aspalatone, a new antithrombotic agent, has been recognized in the neurotoxic model and in the cardiotoxic model in proliminary studies. We examined the specific activity of antiosidnat enzyme in the rat blood following administrations of aspirin, maltol, aspirin together with maltol, salicylmaltol (major metabolite of aspalatone) and aspalatone, respectively. Our assessment showed that salicylmaltol, maltol, aspalatone enhanced antiperoxidative activity. In addition, neither aspirin nor combination of aspirin and maltol, showed any significant effect on the activity of antioxidant enzyme. Because $H_{2}$$O_{2}$ accumulation may stimulate the thrombogenesis in blood, the result suggests that the induction of blood antiperoxidative activity produced by aspalatone may have beneficial effects on the thrombogenesis.

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Antifibrotic Activity of Manassantin B from Saururus chinensis in HSC-T6 Hepatic Stellate Cells

  • Lee, Mi-Kyeong;Yang, Hye-kyung;Yang, Eun-Sun;Kim, Young-Choong;Sung, Sang-Hyun
    • Natural Product Sciences
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    • v.14 no.2
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    • pp.118-121
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    • 2008
  • Manassantin B, a dilignan isolated from Saururus chinensis, significantly inhibited proliferation in HSC-T6 cells in concentration- and time-dependent manners. In addition, treatment of HSC-T6 cells with manassantin B changed cell morphology from flattened myofibroblastic membranous morphology, representing activation state, to slender shape, representing quiescent state. Furthermore, manassantin B effectively reduced collagen content in HSC-T6 cells. These results suggested that manassantin B exerted antifibrotic activity in HSCT6 cells, in part, via inhibition of cell proliferation and decrease of collagen production.

Hepatoprotective activity of methanol extracts of Glinus oppositifolius and Trianthema decandra against paracetamol induced liver damage

  • Gupta, M;Mazumder, UK;Haldar, PK;Manikandan, L;Senthilkumar, GP;Kandar, CC
    • Advances in Traditional Medicine
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    • v.7 no.1
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    • pp.74-78
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    • 2007
  • The plants, Trianthema decandra and Glinus oppositifolius are commonly used by tribal people in India for the treatment of liver diseases. Hepatoprotective activity of methanol extracts of Glinus oppositifolius and Trianthema decandra at the dose of 250 and 500 mg/kg body weight administered orally was evaluated against paracetamol induced liver damage in rats. Biochemical parameters such as serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, serum alkaline phosphatase, bilirubin, total serum protein, lipid peroxide and glutathione content of the liver were estimated to determine liver function and metabolism. From the biochemical observations, it was concluded that methanol extracts of Glinus oppositifolius and Trianthema decandra significantly restored the altered biochemical parameters towards normal condition in paracetamol induced liver damage.

Studies on the Anti-inflammatory Effects of Drymaria cordata Willd

  • Mukherjee, Pulok K.;Mukherjee, Kakali;Bhattacharya, S.;Pal, M.;Saha, B.P.
    • Natural Product Sciences
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    • v.4 no.2
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    • pp.91-94
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    • 1998
  • In folklore medicine Drymaria cordata Willd (Family-Caryophyllaceae) is reported to have laxative and anti-febrile properties along with anti-inflammatory activities. Sikkimis used this plant to treat all these ailments. The anti-inflammatory effect of the methanol extract of D. cordata was investigated against carrageenin, histamine, serotonin, dextran and $PGE_1$ induced rat hind paw oedema. It exhibited significant anti-inflammatory activity against all these phlogestic agents except $PGE_1$ in the order of carrageenin > serotonin > histamine. All these effects were compared with standard drug phenylbutazone in both the acute and chronic experimental models in albino rats.

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Studies on in vivo Wound Healing Activity of Cassia fistula Linn. Leaves (Leguminosae) in Rats

  • Bhakta, T.;Mukherjee, Pulok K.;Mukherjee, Kakali;Pal, M.;Saha, B.P.
    • Natural Product Sciences
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    • v.4 no.2
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    • pp.84-87
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    • 1998
  • Cassia fistula commonly known as Sundali was selected to evaluate its wound healing potentials based on traditional use and literature references. Methanol extract of C. fistula leaves were examined for its wound healing property in the form of an ointment in two types of wound models in rats: i) Excision wound model and ii) Incision wound model. The ointment of the leaf extract of two different concentrations (5% and 10% w/w ointment of leaves extract in simple ointment base) responded significantly in both models of wounds tested. The results were also comparable to that of standard drug, nitrofurazone in terms of wound contraction ability, epithelisation period, tensile strength and regeneration of tissue at wound area.

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Synthesis of Some New Biologically Active Benzothiazole Derivatives Containing Benzimidazole and Imidazoline Moieties

  • Chaudhary, Manish;Pareek, Deepak;Pareek, Pawan K.;Kant, Ravi;Ojha, Krishan G.;Pareek, Arun
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.131-136
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    • 2011
  • Synthesis of N-(1H-benzimidazol-2-yl)-6-substituted-1,3-benzothiazol-2-amines and 6-substituted-N-(4,5-dihydro-1H-imidazol-2-yl)-1,3-benzothiazol-2-amines by the reaction of substituted 2-aminobenzothiazoles with carbon disulphide and methyl iodide followed by the reaction with o-phenylene diamine/ethylene diamine are reported. All the synthesized compounds were characterized by elemental analysis, IR spectra and $^1H$ NMR spectral studies. The potent antibacterial and entomological (antifeedant, acaricidal, contact toxicity and stomach toxicity) activities of the synthesized compounds were investigated.

Ficus racemosa Affords Antihepatotoxic Activity Against Paracetamol-Induced Acute Liver Damage in Rats

  • Mandal, Subhash C.;Maity, Tapan K.;Das, J.;Saha, B.P.;Pal, M.
    • Natural Product Sciences
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    • v.4 no.3
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    • pp.174-179
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    • 1998
  • The effect of oral treatment with Ficus racemosa leaf extract (400 mg/kg for seven days) were studied on hepatic damage induced by paracetamol (750 mg/kg, i.p.) in rats. Biochemical parameter like SGOT, SGPT, serum bilirubin and alkaline phosphatase were estimated to assess liver function. These biochemical observations were supplemented by histopathological examination of liver sections. The activity of extract was also comparable to Neutrosec a known hepatoprotective formulation.

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