Chemical Studies of Formosan Cobra (Naja naja atra) Venom. Part I. Chromatographic Separation of Crude Venom on CM‐Sepadex and Preliminary Characterization of its Components

Lo, Tung‐Bin; Chen, Yee‐Hsiung; Lee, Chen-Yuan

doi:10.1002/jccs.196600004

Cited by 72 publications

(17 citation statements)

References 19 publications

(1 reference statement)

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“…Phosphodiesterase activity was determined by a method modified from Lo et al (29). One hundred microliters of sample was added to an assay mixture containing 0.5 mL of 2.5 mM calcium bis-p-nitrophenylphosphate, 0.3 mL of 0.01 M magnesium sulfate and 0.5 mL of 0.17 M veronal buffer, pH 9.0.…”

Section: Phosphodiesterase Assaymentioning

confidence: 99%

Enzymatic and toxinological activities of Hypnale hypnale (hump-nosed pit viper) venom and its fractionation by ion exchange high performance liquid chromatography

Tan¹,

Sim²,

Gnanathasan³

et al. 2011

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:Hypnale hypnale (hump-nosed pit viper) has been recently identified as one of the medically important venomous snakes in Sri Lanka and on the southwestern coast of India. The characterization of its venom is essential for understanding the pathophysiology of envenomation and for optimizing its management. In the present study, the biological properties of Hypnale hypnale venom and venom fractions obtained using Resource Q ion exchange chromatography were determined. The venom exhibited toxic activities typical of pit viper venom, comparable to that of its sister taxon, the Malayan pit viper (Calloselasma rhodostoma). Particularly noteworthy were its high activities of thrombin-like enzyme, proteases, phospholipase A 2 , L-amino acid oxidase and hyaluronidase. The thrombin-like enzyme was mainly acidic and distributed over several chromatography fractions, indicating its existence in multiple isoforms. The hemorrhagic and necrotic activities of the venom were likely associated with the proteolytic enzyme found mainly in the basic fraction. Phospholipase A 2 and phosphomonoesterase exist in both acidic and basic isoforms, while L-amino acid oxidase and hyaluronidase are highly acidic. The venom clotting activity on fibrinogens showed distinct species specificity in the following increasing order for clotting time: bovine < rabbit < goat < human < horse < < dog, and was comparable to that of C. rhodostoma venom. Its clot formation on human fibrinogen is gradual and prolonged, a phenomenon suggestive of consumptive coagulopathy as a complication observed clinically. At an intramuscular sublethal dose, the venom did not cause acute kidney injury in a rodent model, contrary to the positive control group treated with Daboia russelii venom. Nephrotoxicity may result from higher venom doses in the context of coagulopathy, as a complication provoked by venom hematoxicity.

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Section: Phosphodiesterase Assaymentioning

confidence: 99%

Enzymatic and toxinological activities of Hypnale hypnale (hump-nosed pit viper) venom and its fractionation by ion exchange high performance liquid chromatography

Tan¹,

Sim²,

Gnanathasan³

et al. 2011

J. Venom. Anim. Toxins incl. Trop. Dis

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“…Phosphodiesterase activity was determined using disodium-p-nitrophenol phosphate as substrate (Lo et al, 1966). Reaction mixture containing 0.5 mL of 0.25% venom solution/fraction was added to an assay mixture containing 0.5 mL 0.0025 M sodium-p-nitrophenyl phosphate, 0.3 mL 0.01 M MgSO4, 0.5 mL 0.17 M Tris-HCl buffer (pH 8.0).…”

Section: Neutralization Of Phosphodiesterase Activitymentioning

confidence: 99%

In vitro neutralization of Eachis carinatus and Naja naja venom by Canthium parviflorum and its GC-MS analysis

Shrikanth

Janardhan

2016

Bangladesh J Pharmacol

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Canthium parviflorum is used in the traditional therapy to treat snakebite victims. In the present study, partially purified methanol root extract of C. parviflorum was used for the neutralization of Eachis carinatus and Naja naja venom. The extract inhibited in vitro phosphomonoesterase, phosphodiesterase, acetylcholinesterase, hyaluronidase, protease, phospholipase A2 and 5’nucleotidase activities of both venoms. One of the promising band exhibited neutralization of all the in vitro enzyme activities and was further subjected to GC-MS analysis which revealed the presence of eight active phytocompounds. These phytochemicals might be responsible for in vitro enzyme neutralization. Methanol extract possesses potent active phytochemicals against the site specific toxins and hydrolytic enzymes analyzed.Video Clip of Methodology:<a href="https://youtube.com/v/oSLdFcfaJxU">Phospholipase A2 assay</a>: 1 min 36 sec

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“…Cardiotoxin was isolated from the crude venom by CM-Sephadex column chromatography according to the method of Lo et al (1966), modified by Lee et al (1968).…”

Section: Cardiotoxinmentioning

confidence: 99%

“…Cats, being relatively resibcant to cobra reurotoxin, die of cirulatory failure before the neurotoxin induces peripheral respiratory paralysis (Lee and Peng, 1961;Lee, 1963). Several active components such as neurotoxin, cardilotoxin, phospholipase A, and some proteins having other enzymatic a-Alvfties have been separated from cobra venom (Lo et al, 1966; also see Meldrum, 1965;Boquet, 1966). However, it has not been established as to which component (s) or to what extent these components are responsible for the cardiovascular effects caused by cobra venom.…”

mentioning

confidence: 99%

“…Several active components such as neurotoxin, cardilotoxin, phospholipase A, and some proteins having other enzymatic a-Alvfties have been separated from cobra venom (Lo et al, 1966; also see Meldrum, 1965;Boquet, 1966). However, it has not been established as to which component (s) or to what extent these components are responsible for the cardiovascular effects caused by cobra venom.A cardiotoxic principle was first isolated from the Indian cobra venom by Sarkar (1947) and recently a more purified cardiotoxin has been obtained from the Formosan cobra venom by means of CM-Sephadex column chromatography (Lo et al 1966), and its pharmacological properties have been studied in detail I (Lee et al, 1968). In experiments reported herein, hemodynamic changes produced by cardiotoxJn were analysed in comparison with those prodaced by the whole coora venom, in order to elucidate the role played by cardotoxin in the circulatory effects of cobra venom.…”

mentioning

confidence: 99%

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Studies on Cardiotoxin and Vasoactive Substance Releasing Component(s) of Cobra Venom. Comparison of Hemodynamic Effects of Cardiotoxin with those of Cobra Venom

Lee¹

1971

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The effects of crdiotxr on the cardiovascular system hat been studied in comarison with tboe& 'of crude yeanm of Formosan cobra. The intravenous injection of cardiotozin (1.0 mg/kg) usuaflly produces an iummdate rise in systemic arterial esefolowed by a progressive decline leaing to cardiac arrest, whereas a preiptos 11in systemc arterial pressure is usually obser ve with crue yea 105-1.0 wak). A very marked increase in both pulmonary artery pressure and pulonryvascua resistance is *ioduced foloning the injection of crueo venoma, wfras the increase is far loe conspicuous with cardiotin. Cardiac output as w4 as stroke volus is decreased by both crude veo and cardiotoxi.The early decree in cardiac output with crude veam appears to be resulted frca the marked ibcrease I pulmonary vascular resistance, whereas the progressive decrease leading to circulatc failuz% is due to the direct off ofAndox~ on the heart. Ventricular contractile force is depmessed by an apT c~ltqa smI rcded byc initial augmntation of varying Z V± AI1 01ine.s asSdprui inverted -T, nodal. rhythm, A-V block A-V dissociation, idiovent cLa rhythm a vmsnrcu1&r tachycardia are ao.ere with oardiotxist-as well as zrue venom. V. ca Pren=w in inareased with crude vono probablj as a result of dilation of the ASM * beart, tres no appeclahle change In caval pressure is frwd with oardletada ma lIe" oardlac fumcticss are seriously Impaired at.a later stags. Approved for public releaBe-, distribution unlimited. I:ABSTRACT -The effects of cardiotoxin on the cardiovascular system has been studied in comparison with those of crude venom of Formosan cobra. Tne intravenous injection of cardiotoxin (1. 0 mg/kg) usually produces an. immediate rise in systemic arterial pressure, followed by a progressive decline leading to cardiac arrest, whereas a precipitous fall in systemic arterial pressure is usually observed with crude venom (0. 5 -1. 0 mg/kg). A very marked increase ih both pulmonary artery pressure and pulmonary vascular resistance is produced following the injection of crude venom, whereas the increase is far less conspicuous with cardlotoxin. Cardiac output as well as stroke volume is decreased by both crude venom and cardiotoxin. The early decrease in cardiac output with crude venom appears to be resulted from the marked increase in pulmonary vascular resistance, whereas the progressive decrease leading to circulatory failure is due to the direct effect of cardiotoxin on the heart. ) Ventricular contractile force is depressed'by both crude venom and cardiotoxin, usually preceded by an initial augmentation of varying duration. Abnormal ECG changes such as ST depression, inverted T, nodal rhythm, A-V block, A-V dissociation, idioventricular rhythm and ventricular tachycardiaare observed with cardiotoxin as well as crude venom. V. cava pressure is increased with crude venom probably as a result of dilatation of the right heart, whereas no appreciable change in caval pressure is found with cardiotoxin unless cardiac functions are seriously impaired at a later sta...

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Chemical Studies of Formosan Cobra (Naja naja atra) Venom. Part I. Chromatographic Separation of Crude Venom on CM‐Sepadex and Preliminary Characterization of its Components

Cited by 72 publications

References 19 publications

Enzymatic and toxinological activities of Hypnale hypnale (hump-nosed pit viper) venom and its fractionation by ion exchange high performance liquid chromatography

Enzymatic and toxinological activities of Hypnale hypnale (hump-nosed pit viper) venom and its fractionation by ion exchange high performance liquid chromatography

<i>In vitro</i> neutralization of <i>Eachis carinatus</i> and <i>Naja naja</i> venom by <i>Canthium parviflorum</i> and its GC-MS analysis

Studies on Cardiotoxin and Vasoactive Substance Releasing Component(s) of Cobra Venom. Comparison of Hemodynamic Effects of Cardiotoxin with those of Cobra Venom

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