Local envenomation from the bite of a juvenile false water cobra (Hydrodynastes gigas; Dipsadidae).

The false water cobra (Hydrodynastes gigas) is a non-front-fanged colubroid snake frequently exhibited in zoos, and maintained by amateur collectors. Little detailed documentation regarding the time-course of symptoms development and the consequences of their bites to humans has been published. Reported here is a case of envenoming in a 25 yo male that occurred after the bite of a juvenile H. gigas. The victim was bitten on the fourth digit of the left hand while processing the snake for sex determination, and the snake remained attached to the digit for approximately 30 s; there was no jaw advancement. Within 5 min, intense local pain developed, and at 4hr post bite the entire dorsal aspect of the hand was significantly edematous, The local effects progressed and involved the entire forearm, and the local pain referred to the axillary region. Mild paresthesia and local blanching ("pallor") were noted in the affected digit, but resolved within 7 days. The clinical course in the patient showed that moderate localized symptoms may result from the bite of a juvenile H.gigas.

Non-front-fanged colubroid ("colubrid") snakebites: three cases of local envenoming by the mangrove or ringed cat-eyed snake (Boiga dendrophila; Colubridae, Colubrinae), the Western beaked snake (Rhamphiophis oxyrhynchus; Lamprophiidae, Psammophinae) and the rain forest cat-eyed snake (Leptodeira frenata; Dipsadidae).

CONTEXT: Non-front-fanged colubroid snakes (NFFC; formerly and artificially taxonomically assembled as "colubrids") comprise the majority of extant ophidian species. Although the medical risks of bites by a handful of species have been documented, the majority of these snakes have oral products (Duvernoy's secretions, or venoms) with unknown biomedical properties/unverified functions and their potential for causing harm in humans is unknown. CASE DETAILS: Described are three cases of local envenoming from NFFC bites inflicted respectively by the mangrove or ringed cat-eyed snake (Boiga dendrophila, Colubridae), the Western beaked snake (Rhamphiophis oxyrhynchus, Lamprophiidae) and the rain forest cat-eyed snake (Leptodeira frenata, Dipsadidae). The effects ranged from mild pain, edema and erythema to severe pain, progressive edema, and blistering with slowly resolving arthralgia; there were no systemic effects. DISCUSSION: Although these three taxa occasionally inflict bites with mild to moderate local effects, there is no current evidence of systemic involvement. Two of these cases were reported to one of the authors for medical evaluation, and although verified, thus constitute reliably reported cases, but low-quality evidence. Type-1 local hypersensitivity may contribute to some cases, but most local effects observed or reported in these three cases were consistent with the effects of venom/oral product components.

Preliminary characteristics of the prothrombin converting enzyme from venom of Stephen's banded snake (Hoplocephalus stephensii).

Stephen's banded snake (Hoplocephalus stephensii) is an infrequently encountered Australian elapid species. The crude venom contains coagulant activity and the component responsible is a prothrombin activator requiring factor V for activity. SDS-PAGE of the isolated native protein revealed two bands at 23 and 36 kDa. These findings indicate that the procoagulant is similar to that found in the Australian tiger snake (Notechis scutatus) and thus resembles factor Xa.

Waglerin-1 selectively blocks the epsilon form of the muscle nicotinic acetylcholine receptor.

Neonatal mice resist the lethal effect of Waglerin-1. Because Waglerin-1 blocks the nicotinic acetylcholine receptor of mature end-plates, the appearance of lethality may result from the epsilon- for gamma-subunit substitution. In support of this hypothesis, adult knockout (KO) mice lacking the gene coding for the epsilon-subunit resist the lethal effect of Waglerin-1. In contrast, heterozygous litter mates respond to Waglerin-1 like adult wild-type mice. In vitro application of 1 microM Waglerin-1 inhibited spontaneous miniature end-plate potentials and evoked end-plate potentials of adult wild-type and heterozygous KO mice. Both miniature end-plate potentials and end-plate potentials of neonatal wild-type and adult homozygous KO mice resisted Waglerin-1. Waglerin-1 decreased the end-plate response of adult wild-type mice to iontophoretically applied acetylcholine (ACh) with an IC50 value of 50 nM; 1 microM Waglerin-1 decreased the ACh response to 4 +/- 1% of control for adult heterozygous KO mice. In contrast, 1 microM Waglerin-1 decreased the ACh response to 73 +/- 2% of control for wild-type mice less than 11 days old and had no effect on the ACh response of adult homozygous KO mice. Between 11 and 12 days after birth, the suppressant effect of Waglerin-1 on wild-type end-plate responses to ACh dramatically increased. Waglerin-1 reduced binding of alpha-bungarotoxin to end-plates of adult but not neonatal wild-type mice. These data demonstrate that Waglerin-1 selectively blocks the mouse muscle nicotinic acetylcholine receptor containing the epsilon-subunit.

How to organize and implement health care ergonomics teams.

Using a systems approach to safety and health management through a comprehensive ergonomic program addresses workplace processes, operations and conditions as interdependent systems in order to identify and eliminate or reduce all types of ergonomic hazards to employees. This document examines the way one health care facility improved its ergonomic program using this approach, from early data collection to implementation of the new plan. By creating several teams with different areas of concern, they set out to reduce ergonomic-related injuries in their facility by 10 percent.

Structure-function studies of waglerin I, a lethal peptide from the venom of Wagler's pit viper, Trimeresurus wagleri.

Waglerins are 22-24 residue lethal peptides, found in the venom of Trimeresurus (Tropidolaemus) wagleri. The effects upon lethality and immunoreactivity resulting from structural modifications of these peptides were studied. A synthetic analogue with alanine residues in place of the two half-cystines of native peptide was nontoxic, suggesting that the single intramolecular disulfide bond in waglerins is critical for bioactivity. Substituting glutamic acid for aspartic acid at residue 5 slightly diminished lethality. Analogues containing asparagine instead of aspartic acid at residue 5 and/or a carboxamide- instead of a carboxy-terminus were lethal, demonstrating that neither a negative charge on residue 5 nor on the carboxy-terminus was required for bioactivity. A proteolytic fragment of waglerin I containing residues 6-22 was isolated and proved nontoxic. Therefore, one or more of the first five residues were necessary for bioactivity. Antiserum against waglerin I bound strongly to waglerins I, II, and SL-I, and to various analogues, proteolytic fragments, and chemically modified waglerin I. These findings suggest that the antibodies might be directed mainly against short, linear epitopes, implying an extended conformation for waglerin I.

Properties of Duvernoy's secretions from opisthoglyphous and aglyphous colubrid snakes.

Relatively little attention has been given to the biological properties of Duvernoy's secretions produced by opisthoglyphous and some aglyphous colubrid snakes. A review is presented of literature pertaining to these secretions. Most detailed analyses of Duvernoy's secretions and their biological properties have been performed since the late 1970s. The dispholidines, Dispholidus typus and Thelotornis sp., and the natricines, Rhabdophis tigrinus and R. subminiata, have received the most attention due to the high toxicity of their secretions and their medical importance. These species produce secretions with variably strong prothrombin-activating activity, defibrinating activity, and hemorrhagic potential. Boigines, and natricines other than Rhabdophis, produce secretions of low to moderate toxicity and are variably hemorrhagic and proteolytic. Xenodontines and homalopsines similarly show hemorrhagic potential with low to moderate toxicity. Neurotoxic activity has been reported only from secretions of the boigines, Boiga blandingi and B. irregularis and the xenodontine, Heterodon platyrhinos. These species produce secretions containing postsynaptically acting components. Analyses of some of these secretions have shown that enzymes common to many ophidian venoms such as phospholipases A and L-amino acid oxidase are uncommon in the colubrid secretions studied. This may be due to few studies assaying for multiple enzyme activities and/or the unavailability of many secretion samples for study. Methods of secretion extraction, storage, and assay are discussed. Projected future research and the adaptive implications of Duvernoy's secretions are considered.

Molecular properties and structure-function relationships of lethal peptides from venom of Wagler's pit viper, Trimeresurus wagleri.

Two new lethal peptides (waglerins) were purified from the venom of Trimeresurus wagleri, and sequenced. We found them to be analogs of lethal peptides (waglerins) I and II reported previously (Weinstein et al., Toxicon 29, 227-236, 1991), with an additional Ser-Leu on the amino terminus. Three of the four waglerins were synthesized and the products were chemically and biologically equivalent to the naturally occurring counterparts in venom. Murine i.p. LD50 for synthetic waglerins I, SL-I and II were 0.33, 0.22, and 0.51 mg/kg, respectively. The single, intramolecular disulfide bond in each synthetic peptide formed rapidly in high yield. The reduced (cysteine-containing) forms of the peptides appeared to have significant toxicities, even without prior disulfide bond formation, but synthetic analogs with serine substituted for cysteine were not toxic. The synthetic dimer of waglerin I, formed by two intermolecular disulfide bonds, was not toxic, but rapidly rearranged to lethal, monomeric waglerin I at alkaline pH upon the addition of 5 mM beta-mercaptoethanol. Waglerin I was inactivated by cleavage at Tyr-15 with chymotrypsin.

Analysis of tissue margins of cone biopsy specimens obtained with "cold knife," CO2 and Nd:YAG lasers and a radiofrequency surgical unit.

Analysis of the tissue margins of cone biopsy specimens obtained from 40 patients showed varying degrees of thermal and mechanical artifact at the tissue margins. The least artifact was seen in the tissue margins of specimens obtained with the scalpel ("cold knife"). The amount of thermal damage to biopsies obtained via lasers and the radiofrequency unit varied with the instrument employed. However, the quality of the tissue margins of specimens obtained using a radiofrequency surgical unit equipped with a needle electrode on a "pure cut" setting approached the quality of those obtained with the cold knife in their lack of thermal and mechanical artifact.

A novel peptide toxin from Trimeresurus wagleri acts pre- and post-synaptically to block transmission at the rat neuromuscular junction.

The neuromuscular effects of a peptide toxin (peptide I) from venom of Trimeresurus wagleri were investigated using the rat extensor digitorum longus muscle/peroneal nerve preparation. Sub-micromolar concentrations depressed endplate currents (EPCs) produced in response to nerve stimulation. Since quantal content of EPCs was not altered, it appears that the site of action is post-synaptic. However, higher concentrations (1.4-2.9 microM) also inhibited spontaneous release of transmitter. Nerve stimulation in the presence of peptide I caused 'rundown' of EPC amplitude, evidence that the peptide acts pre-synaptically to interfere with transmitter release. Recovery from this effect occurred within 3-5 min. of washing, but EPC amplitude took 20-30 min. to recover. The dual action of this peptide makes it unusual amongst naturally-occurring toxins, and these data suggest that further investigation of the peptide (and its analogues) could yield new information about neurotransmitter release.

Pathologic changes induced by phospholipase A2 isolated from the venom of Collett's snake, Pseudechis colletti: a light and electron microscopic study.

Pseudechis colletti is an Australian elapid snake with a range limited to central Queensland, Australia. The venom of this snake, as well as that of several other Australian elapids, has been shown to contain a phospholipase A2 (PLA2) which can cause a marked myoglobinuria in mice. Few studies have described the histopathologic and ultrastructural changes that result from myotoxic PLA2-induced damage. Our investigation demonstrated that the isolated PLA2 induced myodegeneration and necrosis in myocardium in a dose-related manner, with subsequent myoglobinuria and myoglobinuric nephropathy.

Facial thermography, basis, protocol, and clinical value.

Computerized electronic telethermography provides clinicians with a reliable evaluation of subtle body surface temperature changes that show underlying physical disorders characterized by pain. The first study population involved 4000 measurements of 100 volunteers at the Medical School of New Jersey. This study of normal volunteers evaluates the validity and reliability of using 0.5 degrees C skin surface temperature difference between opposite sides of the head as a minimum difference standard for recognition of a clinically significant thermographic abnormality. A second study population of over 300 patients with clinically suspected temporomandibular disorder were used. The authors used a standard thermographic protocol procedure that is approved by the Academy of Neuromuscular Thermography. Also discussed in detail are artifactual influences and trigger point detection. The clinical value of this information in the diagnosis and treatment of temporomandibular disorder patients is self evident.

Characterization and amino acid sequences of two lethal peptides isolated from venom of Wagler's pit viper, Trimeresurus wagleri.

Two lethal toxins were isolated from Trimeresurus wagleri venom by fast protein liquid chromatography (molecular sieve) and high performance liquid chromatography (reverse phase). The toxins (termed peptide I and II) had mol. wt of 2504 and 2530, respectively, pIs of 9.6-9.9 and lacked phospholipase A, proteolytic, and hemolytic activity. Lethal peptide I had a murine i.p. LD50 of 0.369 mg/kg, while lethal II had a murine i.p. LD50 of 0.583 mg/kg. Peptide I retained full toxicity after autoclaving at 121 degrees C for 40 min. The lethal activity was found to represent less than 1% of the total venom protein, which was only 62-65% of crude venom. The amino acid sequence of peptide I revealed a proline-rich (over 30% of total sequence) sequence unique among snake venom toxins. Lethal peptide II showed the same sequence except for a second tyrosine in the position of histidine (residue No. 10) in peptide I. The toxin lacked antigenic identity with a number of representative neurotoxins and myotoxins. The crude venom shared at least one antigen with Crotalus scutulatus scutulatus venom. This antigen was not Mojave toxin. The toxin appears symptomatologically suggestive of a vasoactive peptide or neurotoxin.

Lethal potency and fractionation of Duvernoy's secretion from the brown tree snake, Boiga irregularis.

The liquid secretion contained only 15% protein and had relatively low proteolytic activity. The reconstituted crude secretion had a murine i.p. LD50 of 10.33 mg/kg and was not hemorrhagic in doses up to 200 micrograms. Fast Protein Liquid Chromatographic (FPLC) cation exchange analysis of reconstituted crude secretion resulted in resolution of 16 peaks. Lethal activity was identified in three peaks. The major lethal fraction was 12.5% of the secretion protein and had a murine i.p. LD50 of 7.3 mg/kg. A pooled fraction containing two lethal peaks which comprised 9.4% of secretion protein had moderate proteolytic activity and produced myoglobinuria in mice. The fraction had an approximate murine i.p. LD50 of 3.7 mg/kg. Microscopic examination of muscle tissue from mice succumbing to this fraction revealed multifocal myofiber degeneration and necrosis. SDS-PAGE indicated that the major lethal fraction contained three proteins with mol. wts of 12,500, 18,000 and 52,000 and the myotoxic fraction contained two proteins with mol. wts of 14,500 and 17,000. While B. irregularis Duvernoy's secretion has a low lethal index, it does contain a myotoxic fraction with moderate lethal potency. These observations and recent data describing clinical envenomation of several infant patients suggest that large specimens may pose a hazard to infants and small children.

Lethal toxins and cross-neutralization of venoms from the African water cobras, Boulengerina annulata annulata and Boulengerina christyi.

Venoms of the water cobras, Boulengerina, were assayed for lethality, proteolytic activity and protein content. Boulengerina annulata annulata and B. christyi venoms averaged 89% protein and lacked proteolytic activity. The murine i.p. LD50 of B. a. annulata and B. christyi venoms were 0.143 and 0.120 mg/kg, respectively. Polyvalent antivenom produced by the South African Institute of Medical Research neutralized 575 and 200 LD50 of B. a. annulata and B. christyi venoms/ml antivenom, respectively. Cation exchange chromatography resolved four lethal peaks from B. a. annulata venom and six lethal peaks from B. christyi venom. The major lethal peaks (about 12% of total venom protein) were purified further with molecular sieve chromatography and were characterized as 61 (B. a. annulata toxin) and 62 (B. christyi toxin) residue polypeptides with four half-cystines. Elucidation of the complete amino acid sequences indicated that these toxins belonged to the short-chain class of postsynaptic neurotoxins. Short-chain neurotoxins 1 from B. a. annulata and B. christyi had murine i.p. LD50 of 0.052 and 0.083 mg/kg, respectively, and showed over 80% homology with N. nigricollis alpha toxin. Reverse-phase analysis of another peak present in both venoms resolved a toxin that had an N-terminus identical to B. christyi short-chain neurotoxin 1. These fractions also contained toxins readily separable from the short-chain isotoxin by preparative reverse-phase chromatography. Amino acid sequencing of the first 28 residues indicated that both toxins were long-chain neurotoxins with identical N-termini. The LD50 of long-chain neurotoxins 2 from B. a. annulata and B. christyi venoms were 0.086 and 0.090 mg/kg, respectively. The venoms of these little-known elapids have the lowest LD50 of any African proteroglyph studied thus far and have high concentrations of potent postsynaptic neurotoxins.

Antibacterial effects of different snake venoms: purification and characterization of antibacterial proteins from Pseudechis australis (Australian king brown or mulga snake) venom.

Venoms from 30 different snake species were tested in a disc diffusion assay for antibacterial effects against gram-positive and gram-negative bacteria. A number of venoms gave a zone of inhibition against both groups of bacteria, including Aeromonas hydrophila, an important pathogen of reptiles and amphibians. Two antibacterial components from the venom of an Australian elapid, Pseudechis australis (Australian king brown or mulga snake) were purified to homogeneity. The proteins, designated LAO1 and LAO2, had potent antibacterial properties associated with L-amino acid oxidase activity. Both had native and subunit mol. wts of 142,000 and 56,000, respectively. Antibacterial activity correlated with enzymatic activity and was eliminated with catalase. LAO1 and LAO2 had 244 and 113 units of L-amino acid oxidase activity/mg protein, respectively. Compared to tetracycline, a drug of choice for Aeromonas infections in humans, reptiles and amphibians, the in vitro antibacterial effects of LAO1 and LAO2 were respectively 70 and 17.5 times more effective (on a molar basis).

Thermophysiologic anthropometry of the face in Homo sapiens.

Standard clinical thermographic practice has been to regard a side-to-side skin surface temperature difference of 0.5 degrees C or greater as indicative of a clinically significant disorder when such a difference is observed for the contralateral sides of the face. Prior to this time this standard relied upon empiric observation and cumulative experience. This paper provides the first published documentation clearly supportive of this long-standing clinical practice.