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Pharmacological screening of plants recommended by folk medicine as snake venom antidotes; IV. Protection against jararaca venom by isolated constituents.

Fifteen compounds, isolated from plants reputed as snake venom antidotes, belonging to different classes of natural products, were shown to protect mice to a significant degree against the lethal action of the venom of Bothrops jararaca snakes. Administration was by the oral route, one hour prior to envenomation. The substances are nitrogen-free, low-molecular-weight compounds for which some kind of biodynamic activity has previously been reported. The fact that they are mostly trivial, naturally-occurring compounds should explain why plants used as snake-bite antidotes are so widely distributed over the plant kingdom.

Animals↗

[Poisonous snakes of Europe].

Characteristics of poisonous snakes and their toxines are described. The appearance and biology of all European poisonous snakes, eight vipers (family Viperidae) and one opisthoglyph colubride snake (family Colubridae) are given in detail.

Agkistrodon↗

Biological and immunological properties of nerve growth factor from snake venoms.

Homogeneous preparation of nerve growth factor (NGF) was isolated in purity by two steps HPLC fractionation from venoms of snakes belonging to the major families: Crotalidae, Elapidae, and Viperidae. Biological activity of NGF was tested on PC12 cells for neurite outgrowth and molecular weights were determined by PAGE. Antisera raised against NGFs in Balb/C mice. Immunological cross reactivity for antisera was assayed by ELISA and immunoprecipitin tests. HPLC profiles for the venoms from the species belonging to the same family were identical. The biological and immunological properties of NGFs from different species of snake belonging to the same family were also found to be identical. However, NGFs of venoms from different families of snakes showed differences in properties. Neurite outgrowth on PC12 cells due to NGF from the family Elapidae, especially the cobra species, was greater than NGF from the venoms of Crotalidae and Viperidae, with the exception of N. n. nivea which showed poor activity and C. polystictus of Crotalidae family having very good activity.

Animals↗

Processing of pro-tumor necrosis factor-alpha by venom metalloproteinases: a hypothesis explaining local tissue damage following snake bite.

Venom-induced necrosis is a common local debilitating sequela of bites by many vipers, frequently resulting in severe permanent scarring and deformity. Antivenoms are not effective under these circumstances unless administered within a few minutes of the bite; this is unlikely to occur in the rural tropics where most victims take a long time to reach medical care. We have shown that two venom zinc metalloproteinases (jararhagin from Bothrops jararaca venom and a metalloproteinase from Echis pyramidum leakeyi venom) successfully cleaved the recombinant glutathione-S-transferase-tumor necrosis factor-alpha fusion protein (GST-TNF-alpha) substrate to form biologically active TNF-alpha which was shown to be neutralized by ovine TNF-alpha Fab antibodies. This resulted in a reduction of venom-induced necrosis in mice when injected intravenously or intradermally both before and after intradermal injections of E.p.leakeyi venom. A peptidomimetic (POL 647) was also found to inhibit the Echis metalloproteinase, thus preventing the processing of the TNF precursor; this was shown using a TNF-alpha-sensitive cell culture assay and electrophoresis. These observations demonstrate the possible importance of TNF-alpha in the development of the resulting necrotic lesion and leads to the hypothesis that increased levels of venom metalloproteinases following snake bite release active TNF-alpha. This cytokine may contribute to the local necrosis and also induce the production of endogenous matrix metalloproteinases, which in turn generate a positive feedback mechanism resulting in continued cleavage of pro-TNF-alpha. The results indicate that inhibition or neutralization of endogenous TNF-alpha appears to result in a significant reduction in venom-induced necrosis. This could help to explain the clinical observations that treatment of local necrosis following snake bite by antivenom is only minimally successful.

Amino Acid Sequence↗

Lethal factors and enzymes in the secretion from Duvernoy's gland of three colubrid snakes.

Secretion from Duvernoy's gland of the colubrid snakes Malpolon, Spalerosophis, and Thamnophis was obtained by pilocarpine stimulation and tested for lethality and selected enzymatic activities. Pools of secretion from Malpolon and Spalerosophis were fractionated by gel filtration, and several major active fractions were examined. The secretion from Malpolon had an LD50 of 6.5 micrograms/gm in mice; two lethal fractions with LD50's of 2.75 micrograms/gm and 4.5 micrograms/gm were isolated. One of these fractions appears to be a basic phospholipase A with a molecular weight of about 17,000. Spalerosophis had a secretion with an LD50 of 2.75 micrograms/gm in mice; one main lethal fraction with an LD50 of 2.5 micrograms/gm was isolated. Secretion from Thamnophis had an LD50 of 33.3 micrograms/gm; it was not fractionated owing to a paucity of material. At least two of the snake species examined have toxic secretions that may be important during feeding by killing or weakening and helping to subdue the struggling prey.

Animals↗

Nephrotoxic action of rattlesnake and sea snake venoms: an electron-microscopic study.

The fine structure of renal corpuscles and proximal convoluted tubules of the right kidneys of Suiss mice were studied 9 hr after the injection of the venom of Crotalus atrox (Western diamond-back rattlesnake) or the venom of Laticauda semifasciata (broad-banded blue sea snake). Rattlesnake envenomation resulted in several ultrastructural changes in the renal corpuscles and proximal convoluted tubules. Visceral epithelial changes included intracellular oedema, blebbing, vesiculations, the formation of microvillus projections and dilatation of the endoplasmic reticulum and mitochondria. Changes in the parietal epithelium were similar except that no microvillus projections were noted. Mesangiolysis was a consistent finding. Collagenous fibrils were very prominent in the lysed areas of the mesangial cells. Increased numbers of lysosome-related structures were noted in the proximal convoluted tubule cells. Most of the nuclear cisternae of the cells of the renal corpuscles and proximal convoluted tubules were greatly dilated. Sea snake envenomation resulted in focal organellar swelling and focal intracellular oedema of the visceral epithelium. This venom did not affect the cells of the proximal convoluted tubule. The presence of light and dark visceral epithelial cells has been a consistent finding for both the control and the environmated groups.

Animals↗

Conservative and variable regions of homologous snake phospholipases A2 sequences: prediction of the taxon-specific peptides structure.

Homologous amino acid sequences of phospholipases A2 (PLA2) of snakes belonging to the families Elapidae, Viperidae, and Colubridae were considered in order to study the conservative and variable regions location. The PLA2 sequences were divided into two groups (taxons) according to the phylogenetic tree reconstructed from the pair similarity matrix. Results of the intergroup comparison were plotted to facilitate the identification of significant conservative and variable regions. It was shown that the results of the comparison between two phylogenetic groups of snake PLA2 did not much depend on the number of each group representatives and did not markedly change if one of the groups was represented by the single sequence. The knowledge of the number and location of conservative and variable regions and their dependence on the phylogenetic relations between compared taxa may be used to predict a synthetic peptide structure to obtain specific antibodies against PLA2 of one of these taxons. Such prediction is possible if there is a specific region conservative for one taxon but variable for two of them.

Amino Acid Sequence↗

Acute pulmonary edema as a complication of anti-snake venom therapy.

Polyvalent Anti-snake Venom (ASV) is a life-saving antivenin for severe envenomation due to snake bite in India. ASV infusion is occasionally associated with severe allergic reactions, i.e. anaphylaxis and death. We report a rare instance of non-cardiogenic pulmonary edema due to ASV infusion in an eleven years old boy.

Anaphylaxis↗

Snake envenomation. Incidence, clinical presentation and management.

Snake envenomation is a major cause of death and disability in the developing countries, particularly India and Southeast Asia. Species variation in venom components, yield, and lethality leads to quite different clinical presentations and mortality. Venomous snakes are divided into 5 families. Bites of the Viperidae, Crotalidae and Colubridae usually cause primarily local effects and bleeding; the Elapidae most commonly cause neurological symptoms, particularly paralysis; while the Hydrophidae cause paralysis and myolysis. Venoms are complex mixtures of enzymes, peptides and metalloproteins. 26 enzymes have been identified, and 10 of those are found in most venoms. Components have been identified that act as procoagulants, anticoagulants, hyaluronidases, RNases, DNases, postsynaptic toxins and presynaptic toxins. Other peptides induce capillary leak syndrome, haemolysis and shock. The clinical results of envenomation vary widely, and there may be no envenomation with a bite. Syndromes reported include oedema, haemolysis, shock, bleeding, pituitary failure, renal failure, myonecrosis, and combinations of the above. First aid measures that have been proposed include tourniquets, constricting bands, tight crepe bandages, incision and suction, cryotherapy, and high voltage electric shock. None of these has been shown to be effective except usage of a crepe bandage for Australian elapid bite. Tourniquets or cryotherapy, if used for extended periods may lead to gangrene. The most important first aid measure is rapid transport to comprehensive medical care. There is some controversy about medical treatment in the United States, but less in other countries. Supportive measures routinely required include intravenous fluids, tetanus prophylaxis and antibiotics. Anticholinergics may be useful in elapid bite. Intubation and ventilation may be necessary. Unproven surgical approaches include excision of envenomated tissues and fasciotomy. The former is disfiguring, the latter should be reserved for those patients with demonstrated increased intracompartmental pressure. More than 100 antivenins are produced by about 36 laboratories worldwide. The products are effective, but carry a high risk of serum sickness and a lesser risk of anaphylaxis. A more effective and less reactive product is under development.

Africa↗

Venom yields from Australian and some other species of snakes.

The wet and dry venom yields for most Australian native dangerous snakes and a number of non-Australian species are presented. Snakes from the Pseudonaja genus yielded higher than previously published amounts and suggest reconsideration be given to increasing the volume of antivenom in each vial. Higher percentage solids were obtained from venoms from the 4 cobra species (Naja) and Pseudechis genus included in this series.

Animals↗

Molecular dissection of cholinergic binding sites: how do snakes escape the effect of their own toxins?

Snakes have evolved a novel binding site demonstrating selective biorecognition. The snake nicotinic acetylcholine receptor is sensitive to acetylcholine while resistant to the effect of the lethal neurotoxins secreted in their own venom. By subjecting recombinant binding sites to point mutagenesis, biochemical analyses and NMR spectroscopy the binding characteristics of three cholinergic ligands have been measured. The amino acid residue at position 189 has been found to be of particular importance to toxin binding.

Amino Acid Sequence↗

Snake venoms in science and clinical medicine. 1. Russell's viper: biology, venom and treatment of bites.

Russell's viper, Vipera russelli (Shaw), is distributed erratically in 10 south Asian countries and is a leading cause of fatal snake bite in Pakistan, India, Bangladesh, Sri Lanka, Burma and Thailand. In Burma it has been the 5th most important cause of death. Its venom is of great interest to laboratory scientists and clinicians. The precoagulant activity of the venom was used by Macfarlane and others to elucidate the human clotting cascade. Up to 70% of the protein content is phospholipase A2, present in the form of at least 7 isoenzymes. Possible clinical effects of the enzyme include haemolysis, rhabdomyolysis, pre-synaptic neurotoxicity, vasodilatation and shock, release of endogenous autacoids and interaction with monoamine receptors. Russell's viper bite is an occupational hazard of rice farmers throughout its geographical range. Defibrination, spontaneous haemorrhage, shock and renal failure develop with frightening rapidity. In several countries, Russell's viper bite is the commonest cause of acute renal failure. There is a fascinating geographical variation in the clinical manifestations, doubtless reflecting differences in venom composition. Conjunctival oedema is unique to Burma, acute pituitary infarction to Burma and south India, and rhabdomyolysis and neurotoxicity to Sri Lanka and south India. Treatment with potent specific antivenom rapidly controls bleeding and clotting disorders, but may not reverse nephrotoxicity and shock. Causes of death include shock, pituitary and intracranial haemorrhage, massive gastrointestinal haemorrhage and acute tubular necrosis or bilateral renal cortical necrosis. The paddy farmer and the Russell's viper coexist in fragile symbiosis. The snake controls rodent pests but inevitably interacts with man, often with mutually disastrous results.

Acute Kidney Injury↗

Some effects and properties of Duvernoy's gland secretion from Hypsiglena torquata texana (Texas night snake).

The Texas night snake (Hypsiglena torquata texana), a small opisthoglyphic colubrid snake, produces a Duvernoy's secretion which has a relatively low lethal toxicity for mice, but produces pain, hemorrhage, and oedema at the site of injection. The subcutaneous LD50 of Hypsiglena torquata texana Duvernoy's secretion in Swiss-Webster mice was 26 mg/kg. Local hemorrhaging occurred at doses as low as 2.0 micrograms/mouse, while marked hemorrhage and oedema resulted at doses greater than 200 micrograms/mouse. The dry secretion contained 77% protein, and SDS-PAGE electrophoresis revealed the presence of seven major proteinaceous components.

Animals↗

Development of a rapid and sensitive enzyme-linked immunosorbent assay (ELISA) for measuring venom antigens after an experimental snake bite.

We describe a new ELISA which allows the measurement of the concentration of venom antigens in whole blood. The assay can be performed in less than 20 min and requires a 200 microliters sample of blood. It allows the accurate evaluation of concentrations of Vipera ammodytes venom in quantities smaller than 1 ng/ml of blood. Using this ELISA, we were able to follow in rabbits the kinetics of experimental envenomation with non-lethal doses of venom. This ELISA was also used to measure post mortem the level of venom antigens in various tissues such as liver, kidney, muscles and abdominal serosity of a rabbit. The method, which might be adapted to measure envenomation by other snake species, seems to be sufficiently rapid and sensitive to allow routine evaluation of the gravity of a snake bite in humans and to estimate the efficacy of immunotherapy.

Animals↗

Neutralization of hemorrhagic snake venoms by sera of Trimeresurus flavoviridis (Habu), Herpestes edwardsii (mongoose) and Dinodon semicarinatus (Akamata).

The sera of T. flavoviridis (Habu), H. edwardsii (mongoose) and D. semicarinatus (Akamata, non-venomous snake) were tested for their capacity to neutralize 28 species of hemorrhagic snake venoms in vitro. The sera of these animals neutralized a variety of hemorrhagic venoms, suggesting a common structure for a hemorrhagic factor and a similar mechanism of neutralization for the antihemorrhagic factor in the sera. The serum of T. flavoviridis neutralized the lethal toxicity of the T. flavoviridis venom but could not neutralize those of the other hemorrhagic venoms at all. The sera of H. edwardsii and D. semicarinatus did not inhibit the activity of all hemorrhagic venoms.

Animals↗

Hemostatic changes due to the venom gland extract of the red-necked keelback snake (Rhabdophis subminiatus).

After a bite by the aglyphous red-necked keelback snake Rhabdophis subminiatus a complete defibrinogenation syndrome with severe hemorrhagic diathesis developed in a 25-year-old man. In vitro studies showed that the venom gland extract of the snake contains a very active prothrombin (Factor II) activator. The thrombin generated is inhibited neither by antithrombin III nor the antithrombin-III-heparin complex. The venom gland extract stimulated also the tissue plasminogen activator; however, it did not cause direct activation of plasminogen, protein C, Factor X or direct degradation of fibrinogen.

Adult↗

Screening of snake venoms for neurotoxic and myotoxic effects using simple in vitro preparations from rodents and chicks.

Eight snake venoms designated by the WHO as International Reference Venoms, and one additional venom were assessed for neurotoxic and myotoxic effects in vitro using the chick biventer cervicis and the rat and mouse phrenic nerve-diaphragm preparations. The objective was to determine whether any of the preparations could be used to detect evidence of neurotoxic or myotoxic activity prior to a more detailed examination. Bungarus multicinctus venom at concentrations above 1 microgram ml-1 selectively blocked neuromuscular transmission, with no direct effect on muscle fibres. Naja naja kaouthia and Notechis scutatus venoms selectively blocked neuromuscular transmission at low concentrations, but at higher concentrations both venoms caused direct effects on skeletal muscle resulting in contractures, loss of tension following direct stimulation and a loss in sensitivity to elevated [K+]0. Vipera russelli (Thailand) venom also blocked neuromuscular transmission but it was less potent than the venoms of B. multicinctus, N. n. kaouthia and N. scutatus. It also caused contractures in the chick biventer cervicis muscle. The venoms of Echis carinatus (Iran and Mali), Crotalus atrox, Bothrops atrox asper and Trimeresurus flavoviridis had limited neuromuscular blocking activity, and most of these venoms blocked [K+]0 and cholinoceptor stimulation in the chick muscle. Although both chick and rodent muscles allowed the assessment of neurotoxic and myotoxic activity, the chick biventer cervicis was simpler and more robust in use than either of the rodent phrenic nerve-diaphragm preparations. We propose that the chick biventer cervicis muscle could be used as a standard preparation for the screening of snake venoms for neurotoxic and myotoxic effects, and that it may be possible to use this preparation as a means to check that antivenoms can neutralize neurotoxic and direct myotoxic actions of venoms.

Animals↗

The use of the chick biventer cervicis preparation to assess the protective activity of six international reference antivenoms on the neuromuscular effects of snake venoms in vitro.

The protective activity of some antivenoms on the neuromuscular activity in vitro of six snake venoms was established in order to test the feasibility of using a simple isolated nerve-muscle preparation to compare different antivenoms. Six venoms designated as International Reference Venoms by the World Health Organization (WHO) were used: Echis carinatus (Iran), Echis carinatus (Mali), Naja naja kaouthia, Notechis scutatus, Trimeresurus flavoviridis, and Vipera russelli (Thailand). The chick biventer cervicis nerve-muscle preparation was used to detect neurotoxic and myotoxic effects. The ratio of the amount of antivenom needed to neutralize a dose of venom (w/w) was calculated in order to quantify the potencies of the antivenoms. The antivenom to venom ratios were 1000 for Echis carinatus (Mali) and for Notechis scutatus; 100 for Naja naja kaouthia and Vipera russelli; and 10 for Echis carinatus (Iran) and Trimeresurus falvoviridis. It is concluded that in vitro assays using the chick biventer cervicis preparation could be used to compare the relative potencies of different antivenoms at neutralizing myotoxic and neurotoxic toxins in snake venoms.

Acetylcholine↗