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Suk-ho Choi - One of the best experts on this subject based on the ideXlab platform.

  • Isolation and Characterization of a 32-kDa Fibrinolytic Enzyme (FE-32kDa) from Gloydius blomhoffii siniticus Venom: Fibrinolytic Enzyme from Gloydius blomhoffii siniticus Venom
    Journal of pharmacopuncture, 2014
    Co-Authors: Joung-yoon Kim, Seung-bae Lee, Ki-rok Kwon, Suk-ho Choi
    Abstract:

    Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent.

  • Isolation from Gloydius blomhoffii siniticus Venom of a Fibrin(ogen)olytic Enzyme Consisting of Two Heterogenous Polypeptides.
    Journal of pharmacopuncture, 2013
    Co-Authors: Suk-ho Choi, Seung-bae Lee
    Abstract:

    Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfatepolyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Res sults: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. s initicus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than 20 μ g of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as

  • Characterization of Fibrinolytic Proteases from Gloydius blomhoffii Siniticus Venom
    Journal of Acupuncture and Meridian Studies, 2012
    Co-Authors: Suk-ho Choi
    Abstract:

    Objectives : This study was undertaken to identify fibrinolytic proteases from Gloydius blomhoffii siniticus venom and to characterize a major fibrinolytic protease purified from the venom. Methods : The venom was subjected to chromatography using columns of Q-Sepharose and Sephadex G-75. The molecular weights of fibrinolytic proteases showing fibrinolytic zone in fibrin plate assay were determined in SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) The effects of inhibitors and metal ions on fibrinolytic protease and the proteolysis patterns of fibrinogen, gelatin, and bovine serum albumin were investigated. Results : 1) The fibrinolytic fractions of the three peaks isolated from Gloydius blomhoffii siniticus venom contained two polypeptides of 46 and 59 kDa and three polypeptides of 32, 18, and 15 kDa and a major polypeptide of 54 kDa, respectively. 2) The fibrinolytic activity of the purified protease of 54 kDA was inhibited by metal chelators, such as EDTA, EGTA, and 1,10-phenanthroline, and disulfhydryl-reducing compounds, such as dithiothreitol and cysteine. 3) Calcium chloride promoted the fibrinolytic activity of the protease, but mercuric chloride and cobalt(II) chloride inhibited it. 4) The fibrinolytic protease cleaved preferentially A-chain and slowly B-chain of fibrinogen. It also hydrolyzed gelatin but not bovine serum albumin. Conclusions : The Gloydius blomhoffii siniticus venom contained more than three fibrinolytic proteases. The major fibrinolytic protease was a metalloprotease which hydrolyzed both fibrinogen and gelatin, but not bovine serum albumin.

Choi Suk-ho - One of the best experts on this subject based on the ideXlab platform.

  • Isolation and Characterization of a 32-kDa Fibrinolytic Enzyme (FE-32kDa) from Gloydius blomhoffii siniticus Venom
    Korean Pharmacopuncture Institute, 2014
    Co-Authors: Kim Joung-yoon, Lee Seung-bae, Kwon Ki Rok, Choi Suk-ho
    Abstract:

    Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as Fe2+ and Hg2+ inhibited the fibrin hydrolysis completely, but Zn2+ enhanced it. FE-32kDa hydrolyzed α-chain but did not hydrolyze β-chain and γ-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of 2 μg. Conclusions: FE-32kDa is a α-fibrin(ogen)olytic metalloprotease that requires Zn2+ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture

  • Isolation from Gloydius blomhoffii siniticus Venom of a Fibrin(ogen)olytic Enzyme Consisting of Two Heterogenous Polypeptides
    Korean Pharmacopuncture Institute, 2013
    Co-Authors: Choi Suk-ho, Lee Seung-bae
    Abstract:

    Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfatepolyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. siniticus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than 20 μg of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as indicating its fibrinolytic activity. The fibrinolytic activity was inhibited completely by phenylmethanesulfonylfluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) and partially by thiothreitol and cysteine. Metal ions such as Hg2+ and Fe2+ inhibited the fibrinolytic activity completely, but Mn2+ did not. FE-27kDa preferentially hydrolyzed α-chain of fibrinogen and slowly hydrolyzed β-chain, but did not hydrolyze γ-chain. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into polypeptides with molecular weights of more than 45 kDa. A dosage of more than 10 μg of FE- 27kDa per mouse was required to induce hemorrhage beneath the skin. Conclusion: FE-27kDa was a serine proteinase consisting of two heterogeneous polypeptides, hydrolyzed fibrin, fibrinogen, and gelatin, and caused hemorrhage beneath the skin of mouse. This study suggests that the potential of FE-27kDa as pharmacopuncture agent should be limited due to low fibrinolytic activity and a possible side effect of hemorrhage

Lee Seung-bae - One of the best experts on this subject based on the ideXlab platform.

  • Isolation and Characterization of a 32-kDa Fibrinolytic Enzyme (FE-32kDa) from Gloydius blomhoffii siniticus Venom
    Korean Pharmacopuncture Institute, 2014
    Co-Authors: Kim Joung-yoon, Lee Seung-bae, Kwon Ki Rok, Choi Suk-ho
    Abstract:

    Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as Fe2+ and Hg2+ inhibited the fibrin hydrolysis completely, but Zn2+ enhanced it. FE-32kDa hydrolyzed α-chain but did not hydrolyze β-chain and γ-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of 2 μg. Conclusions: FE-32kDa is a α-fibrin(ogen)olytic metalloprotease that requires Zn2+ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture

  • Isolation from Gloydius blomhoffii siniticus Venom of a Fibrin(ogen)olytic Enzyme Consisting of Two Heterogenous Polypeptides
    Korean Pharmacopuncture Institute, 2013
    Co-Authors: Choi Suk-ho, Lee Seung-bae
    Abstract:

    Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfatepolyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. siniticus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than 20 μg of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as indicating its fibrinolytic activity. The fibrinolytic activity was inhibited completely by phenylmethanesulfonylfluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) and partially by thiothreitol and cysteine. Metal ions such as Hg2+ and Fe2+ inhibited the fibrinolytic activity completely, but Mn2+ did not. FE-27kDa preferentially hydrolyzed α-chain of fibrinogen and slowly hydrolyzed β-chain, but did not hydrolyze γ-chain. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into polypeptides with molecular weights of more than 45 kDa. A dosage of more than 10 μg of FE- 27kDa per mouse was required to induce hemorrhage beneath the skin. Conclusion: FE-27kDa was a serine proteinase consisting of two heterogeneous polypeptides, hydrolyzed fibrin, fibrinogen, and gelatin, and caused hemorrhage beneath the skin of mouse. This study suggests that the potential of FE-27kDa as pharmacopuncture agent should be limited due to low fibrinolytic activity and a possible side effect of hemorrhage

Seung-bae Lee - One of the best experts on this subject based on the ideXlab platform.

Motohide Takahashi - One of the best experts on this subject based on the ideXlab platform.

  • surveillance of the clinical use of mamushi Gloydius blomhoffii antivenom in tertiary care centers in japan
    Japanese Journal of Infectious Diseases, 2011
    Co-Authors: Toru Hifumi, Akihiko Yamamoto, Kazunori Morokuma, Nobuaki Kiriu, Tomoko Ogasawara, Hiroshi Kato, Junichi Inoue, Yuichi Koido, Eiju Hasegawa, Motohide Takahashi
    Abstract:

    We report the results of the first large-scale questionnaire surveillance on the clinical use of pit viper antivenom in tertiary care centers in Japan. The questionnaire surveillance was conducted over a period of 3 years (April 2006 to March 2009). Completed questionnaires were received from the tertiary care centers of 108 (49.3%) medical institutions. In that period, 574 cases of pit viper bites, including 2 severe cases, were reported. Antivenom was administered in 44% of the cases of pit viper bites, and of these cases, 2.4% had adverse reactions but no severe symptoms. Approximately half of the clinicians indicated that antivenom was effective. Antivenom was recognized to be safe; however, the remarkable finding was that although the severity of treated cases was unclear, some clinicians reported using cepharanthine as the first choice of treatment for pit viper bites.

  • surveillance of the clinical use of mamushi Gloydius blomhoffii antivenom in tertiary care centers in japan
    Japanese Journal of Infectious Diseases, 2011
    Co-Authors: Toru Hifumi, Akihiko Yamamoto, Kazunori Morokuma, Nobuaki Kiriu, Tomoko Ogasawara, Hiroshi Kato, Junichi Inoue, Yuichi Koido, Eiju Hasegawa, Motohide Takahashi
    Abstract:

    Human Vaccine Production Department, The Chemo-Sera-Therapeutic Research Institute(KAKETSUKEN), Kumamoto 860-8568, Japan(Received February 10, 2011. Accepted June 16, 2011)SUMMARY: We report the results of the first large-scale questionnaire surveillance on the clinical useof pit viper antivenom in tertiary care centers in Japan. The questionnaire surveillance was conductedover a period of 3 years (April 2006 to March 2009). Completed questionnaires were received from thetertiary care centers of 108 (49.3z) medical institutions. In that period, 574 cases of pit viper bites, in-cluding 2 severe cases, were reported. Antivenom was administered in 44z of the cases of pit viperbites, and of these cases, 2.4z had adverse reactions but no severe symptoms. Approximately half ofthe clinicians indicated that antivenom was effective. Antivenom was recognized to be safe; however,the remarkable finding was that although the severity of treated cases was unclear, some cliniciansreported using cepharanthine as the first choice of treatment for pit viper bites.INTRODUCTIONJapanese mamushi, Gloydius blomhoffii, a species ofpit viper distributed throughout Japan excluding Ryu-kyu Islands, is sighted from spring to autumn. It is im-portant that many people are bitten by this pit viper inthe mountains and fields of rural Japan. The annualnumber of pit viper bites remains unclear because thereis currently no system to report pit viper bites to thepublic health department in Japan. Some reports esti-matethenumberofpitviperbitestobe1,000with10deaths annually (1).Fatalities due to pit viper bites are generally low, butsevere cases involving cardiac, pulmonary, and/or renaldysfunction can be lethal (2–4). These symptoms arecaused by the snake's venom, which has lethal andhemorrhagic activities (5). Passive immunizationagainst the venom is crucial for the clinical treatment ofbites. Antivenom can neutralize both the hemorrhagicand lethal activities of venom. However, since they arederived from horse serum, these exogenous serumproducts frequently cause shock and anaphylaxis (6). Asatisfactory treatmentstrategy hasbeen proposedon thebasis of the progress of symptoms following pit viperbites (1). An essential and rationalized therapy for se-vere cases of pit viper bites is rapid intravenous adminis-tration of antivenom. Therefore, antivenom should beadministered to the snakebite victim safely and quickly.The annual antivenom production in Japan is 3,000doses, which is gradually decreasing because of thelimited opportunity for use (7).Any snakebite victim should immediately visit ahospital's emergency department, unless the snake hasbeen positively identified as nonvenomous, because ofthe potentiallethaleffects ofsnakevenom.Weconduct-ed this survey to elucidate the number of snakebite casesand related therapy in tertiary care centers in Japan.This is the first large-scale questionnaire surveillance onthe clinical use of pit viper antivenom in tertiary carecenters in Japan.We received reports of 574 cases of pit viper bites, in-cluding 2 severe cases, from tertiary care centers of 67medical institutions. Antivenom was administered in44z of pit viper bite cases, and adverse reactions to theserum were reported in only 2.4z of cases, with no se-vere symptoms.METHODSWe used a newly designed questionnaire to survey theclinical use of pit viper antivenom at all 219 tertiary carecenters in Japan. The questionnaire was sent in October2009. The completed questionnaires were collected wi-thin 3 months. The content of the questionnaire is pro-vided in Table 1. The questionnaire surveillance wasconducted for a period of 3 years (April 2006 to March2009).RESULTSCompleted questionnaires were received from the ter-tiary care centers of 108 (49.3z) medical institutionsthat reported 574 cases of pit viper bites, including 3cases of complications. Among the centers thatresponded, 67 reported having treated cases of pit viper

  • Standardization of Regional Reference for Mamushi (Gloydius blomhoffii) Antivenom in Japan, Korea, and China
    Japanese journal of infectious diseases, 2006
    Co-Authors: Tadashi Fukuda, Kazunori Morokuma, Masaaki Iwaki, Seung Hwa Hong, Zhu Wei, Kunio Ohkuma, Lei Dianliang, Yoshichika Arakawa, Motohide Takahashi
    Abstract:

    The mamushi (Gloydius blomhoffii) snakes that inhabit Japan, Korea, and China produce venoms with similar serological characters to each other. Individual domestic standard mamushi antivenoms have been used for national quality control (potency testing) of mamushi antivenom products in these countries, because of the lack of an international standard material authorized by the World Health Organization. This precludes comparison of the results of product potency testing among countries. We established a regional reference antivenom for these three Asian countries. This collaborative study indicated that the regional reference mamushi antivenom has an anti-lethal titer of 33,000 U/vial and anti-hemorrhagic titer of 36,000 U/vial. This reference can be used routinely for quality control, including national control of mamushi antivenom products.