Gomes A

Snake venom neutralization by Indian medicinal plants (Vitex negundo and Emblica officinalis) root extracts

The methanolic root extracts of Vitex negundo Linn. and Emblica officinalis Gaertn. were explored for the first time for antisnake venom activity. The plant (V. negundo and E. officinalis) extracts significantly antagonized the Vipera russellii and Naja kaouthia venom induced lethal activity both in in vitro and in vivo studies. V. russellii venom-induced haemorrhage, coagulant, defibrinogenating and inflammatory activity was significantly neutralized by both plant extracts. No precipitating bands were observed between the plant extract and snake venom. The above observations confirmed that the plant extracts possess potent snake venom neutralizing capacity and need further investigation.

Viper venom-induced inflammation and inhibition of free radical formation by pure compound (2-hydroxy-4-methoxy benzoic acid) isolated and purified from anantamul (Hemidesmus indicus R.Br) root extract

The present investigation explored the possible venom neutralizing effect of a pure compound (2-hydroxy-4-methoxy benzoic acid) isolated and purified from the methanolic root extract of Hemidesmus indicus R.Rr. 2-OH-4-MeO benzoic acid possessed potent anti-inflammatory, antipyretic and antioxidant properties. The compound effectively neutralized inflammation induced by Vipera russelli venom in male albino mice and reduced cotton pellet-induced granuloma in rats. The compound produced a significant fall in body temperature in yeast-induced pyrexia in rats but did not change the normothermic body temperature. The compound effectively neutralized viper venom-induced changes in serum phosphatase and transaminase activity in male albino rats. It also neutralized free radical formation as estimated by TBAPS and superoxide dismutase activities. The antisnake venom activity of the pure compound is partly mediated through the above physiological process.

Isolation, purification and partial characterization of viper venom inhibiting factor from the root extract of the Indian medicinal plant sarsaparilla (Hemidesmus indicus R.Br.)

An organic acid, isolated and purified from the root extract of an Indian medicinal plant sarsaparilla Hemidesmus indicus R. Br, possessed viper venom inhibitory activity. The compound (designated HI-RVIF) was isolated by solvent extraction, silica gel column chromatography and thin layer chromatography, and was homogeneous in nature. The white needle-shaped crystals were soluble in water, methanol and chloroform and had a melting point of 155-158 degrees C and lambda max 260 nm. Spectral analysis confirmed the presence of a benzene ring, methoxy group, and hydroxyl group; the mol. wt of the compound was 168. HI-RVIF significantly antagonized viper venom-induced lethal, haemorrhagic, coagulant and anticoagulant activity in experimental rodents.

Adjuvant effects and antiserum action potentiation by a (herbal) compound 2-hydroxy-4-methoxy benzoic acid isolated from the root extract of the Indian medicinal plant 'sarsaparilla' (Hemidesmus indicus R. Br.)

The adjuvant effect and antiserum potentiation of a compound 2-hydroxy-4-methoxy benzoic acid were explored in the present investigation. This compound, isolated and purified from the Indian medicinal plant Hemidesmus indicus R. Br, possessed antisnake venom activity. Rabbits immunized with Vipera russellii venom in the presence and absence of the compound along with Freunds complete adjuvant, produced a precipitating band in immunogel diffusion and immunogel electrophoresis. The venom neutralizing capacity of this antiserum showed positive adjuvant effects as evident by the higher neutralization capacity (lethal and hemorrhage) when compared with the antiserum raised with venom alone. The pure compound potentiated the lethal action neutralization of venom by commercial equine polyvalent snake venom antiserum in experimental models. These observations raised the possibility of the use of chemical antagonists (from herbs) against snake bite, which may provide a better protection in presence of antiserum, especially in the rural parts of India.

Viper venom neutralization by Indian medicinal plant (Hemidesmus indicus and Pluchea indica) root extracts

The methanol root extracts of Hemidesmus indicus R. Br. and Pluchea indica (Less) were explored for the first time for neutralization of snake venom (Vipera russellii) activity. The H. indicus and P. indica root extracts significantly neutralized the viper venom‐induced lethality and haemorrhagic activity in albino rat and mouse. Venom‐induced coagulant and anticoagulant activity was also antagonized by both the extracts. No precipitating bands were observed between the plant extract and polyvalent snake venom antiserum. Maximum neutralization was achieved by H. indicus root extract. These observations confirmed that certain Indian medicinal plants possess significant snake venom neutralizing capacity and need further examination for their active constituents.

Occurrence of a unique protein toxin from the Indian King Cobra (Ophiophagus hannah) venom

A unique (lethal-cardiotoxic-hemorrhagic) protein toxin (Toxin CM55) was isolated and purified from Indian King Cobra (Ophiophagus hannah) venom by CM-sephadex ion exchange chromatography and reverse phase HPLC. The purified toxin had an SDS-molecular weight of 22 +/- 0.5 kD. UV absorption spectra of Toxin CM55 showed a peak at 280 nm, whereas when excited at 280 nm fluorescence, Toxin CM55 showed an E(max) at 333.4 nm. Toxin CM55 had an LD(50) of 28.28 microg/20 g (i. v.) in albino mice. The cardiotoxic action of the toxin was established on isolated guinea pig/rabbit heart and guinea pig auricle. In rats, Toxin CM55 caused ECG abnormalities including widened QRS complex and monomorphic ventricular tachycardia suggesting that the possible site of action of Toxin CM55 was the ventricle. Toxin CM55 produced significant vasoconstriction on peripheral blood vessels. It produced significant contraction of isolated guinea pig ileum, rat fundus and rat uterus, which was completely antagonised by methysergide. The toxin was found to release a significant amount of serotonin from rabbit platelets. Toxin CM55 produced cutaneous hemorrhage in albino mice, which was also produced in reserpine and p-chloro phenylalanine pretreated animals. Rabbit antiserum was raised against Toxin CM55, which gave prominent bands in immunogel diffusion and immunoelectrophoresis. The antiserum provided 2 LD(50) protection against Toxin CM55-induced lethality in mice and also neutralised 3 MHD hemorrhagic dose of the toxin.

Anti arthritic and anti inflammatory activity of a cytotoxic protein NN-32 from Indian spectacle cobra (Naja naja) venom in male albino rats

The anti arthritic and anti inflammatory activity of NN-32, a cytotoxic protein from Indian spectacle cobra snake (Naja naja) venom has been studied in Freunds complete adjuvant (FCA) induced arthritis and carrageenan induced anti inflammatory model. NN-32 treatment showed significant decrease in physical and urinary parameters, serum enzymes, serum cytokines levels as compared to arthritic control group of rats. NN-32 treatment recovered carrageenan induced inflammation as compared to control group of rats. The findings showed that the cytotoxic protein NN-32 shares anti arthritic and anti inflammatory activity and thus NN-32 may target complex pathophysiological processes like cancer- arthritis-inflammation.

A sleep inducing factor from common Indian toad (Bufo melanostictus, Schneider) skin extract.

Bufo melanostictus (common Indian toad) acquire different bioactive substances in their skin during their life-time in wide ecological habitat. Earlier investigation from this laboratory revealed that toad (B. melanostictus) skin extract (TSE) posses different bioactive compounds of different diversity (Das, M., Auddy, B. and Gomes, A., 1996. Pharmacological study of the toad skin extract on experimental animals. Indian J. Pharmacol. 28, 72-76). Among these sleep induction and sleep potentiation indicated the possibility of sleep inducing factor(s) in TSE. One such sleep inducing factor (SIF) was isolated and purified by neutral alumina column chromatography followed by HPLC. Spectroscopy (UV, IR, FAB-MASS) study indicated that the sleep inducing factor was a 880 Dalton conjugated aromatic compound with a hydroxyl and carbonyl functional group. Biological study showed that SIF produced no lethality in male albino mice upto the dose of 8 mg/kg, i.v. Cyproheptadine antagonised SIF induced contraction of isolated smooth muscle indicating histamine/serotonin receptor mediated action of SIF. EEG studies showed that SIF increased sleep and decreased awakening condition of freely moving rats. Biochemical studies showed that SIF produced significant alteration of brain biogenic amine levels, monoamine oxidase (MAO) and tryptophan hydroxylase (TH) activity. This may be the reason of SIF induced sleep, although the SIF induced sleep mechanism needs further detail investigation.

Isolation of a toxin from jellyfish Acromitus rabanchatu and its effect on skeletal muscle

A toxin (T-Ar) from the crude extract of jellyfish Acromitus rabanchatu was purified by (NH4)2SO4 fractionation followed by ion-exchange chromatography on DEAE-cellulose column. The mol. wt of T-Ar was found to be about 182,000. T-Ar reversibly blocked the twitch response of rat diaphragm muscle stimulated either indirectly or directly. It also reversibly blocked the twitch responses of chick biventer-cervicis muscle preparations. T-Ar failed to alter the contractures of the denervated rat diaphragm muscle preparation induced by acetylcholine, potassium chloride and caffeine. Similarly, acetylcholine and carbachol-induced contractures on chick biventer-cervicis muscle preparation were not influenced by T-Ar. The T-Ar-induced blockade of twitch responses of rat phrenic nerve-diaphragm was enhanced in high Na+ or low Ca2+ medium, and reduced in low Na+ or high Ca(2+)-containing medium. The results suggest that the toxin (T-Ar) possesses a direct paralysing effect on the muscle membrane.

Therapeutic potential of krait venom

Kraits belong to Elapideae and are widely distributed in East and South-East Asian countries. Krait venom possesses neurotoxins, membrane toxins, cardiotoxins, three finger toxins, metalloproteinases, cholinesterases, L-amino acid oxidases and serine proteases. The therapeutic potential of krait venom in pathophysiological conditions such as microbial and parasitic infections, cancer, arthritis, inflammation and blood coagulation disorder is discussed in this review. More intensive new research ventures are required to establish the therapeutic potential of krait venom in complex and emerging diseases.