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Injection sclerotherapy for spider telangiectasias: a 20-year experience with sodium tetradecyl sulfate.

A 20-year experience using sodium tetradecyl sulfate (Sotradecol, Elkins-Sinn, Inc., Cherry Hill, NJ) for the injection treatment of spider telangiectasias is reviewed. Of 1426 legs injected, 882 were treated with a 1%-solution injected directly into the spider veins, and 264 were injected directly with a 1/3%-solution. With the 1/3%-solution, the remaining 280 legs had the blue veins feeding the spider telangiectasias injected rather than the spider itself. When injected directly into the spider veins, the lesser concentration gave fewer ulcerations and less pigmentation, but equal treatment results. Injecting the subcutaneous feeder veins appears to provide more permanent clearing of the lesions with fewer recurrences and no side effects. No allergic or other major complications occurred with the injection treatments.

Adolescent↗

The significance of relatedness and gene flow on population genetic structure in the®subsocial spider Eresus cinnaberinus (Araneae: Eresidae)

Interdemic selection, inbreeding and highly structured populations have been invoked to explain the evolution of cooperative social behaviour in the otherwise solitary and cannibalistic spiders. The family Eresidae consists of species ranging from solitary and intermediate subsocial to species exhibiting fully cooperative social behaviour. In this study we, in a hierarchical analysis, investigated relatedness of putative family clusters, inbreeding and population genetic structure of the subsocial spider Eresus cinnaberinus. Five hierarchical levels of investigation ranging from large scale genetic structure (distances of 250 and 50 km level 1 and 2) over microgeographic structure (20 km2 and 4 km2, level 3 and 4) to a single hill transect of 200 m (level 5) were performed. The purpose of level 5 was two-fold: (1) to investigate the relatedness of putative family groups, and (2) to evaluate the influence of both family living and sampling design on higher level estimates. Relatedness estimates of putative family groups showed an average relatedness of R=0.26. There was no indication of inbreeding. In contrast to social spiders, genetic variation was abundant, Heapproximately0.10. The population genetic structure was intermediate between social and asocial spiders. Genetic variance increased continually across hierarchical levels. Family structured neighbourhoods biased differentiation estimates among level 5 samples (FST=0.04) and level 3 and 4 samples (0.07 0, was caused by disjunct sampling from separate neighbourhoods. Larger scale samples were highly differentiated 0.12<FST<0.26, depending on level and sampling design. Due to a distance effect family living did not influence estimates of the higher level 1. Although the dispersing sex among social spiders and the subsocial E. cinnebarinus differ, females versus males, female behaviour of both sociality classes lead to high genetic variance.Copyright 1998 The Linnean Society of London

Journal Article↗

Pharmacologically active spider peptide toxins.

Advances in mass spectrometry and peptide biochemistry coupled to modern methods in electrophysiology have permitted the isolation and identification of numerous novel peptide toxins from animal venoms in recent years. These advances have also opened up the field of spider venom research, previously unexplored due to methodological limitations. Many peptide toxins from spider venoms share structural features, amino acid composition and consensus sequences that allow them to interact with related classes of cellular receptors. They have become increasingly useful agents for the study of voltage-sensitive and ligand-gated ion channels and the discrimination of their cellular subtypes. Spider peptide toxins have also been recognized as useful agents for their antimicrobial properties and the study of pore formation in cell membranes. Spider peptide toxins with nanomolar affinities for their receptors are thus promising pharmacological tools for understanding the physiological role of ion channels and as leads for the development of novel therapeutic agents and strategies for ion channel-related diseases. Their high insecticidal potency can also make them useful probes for the discovery of novel insecticide targets in the insect nervous system or for the development of genetically engineered microbial pesticides.

Amino Acid Sequence↗

Channels produced by spider venoms in bilayer lipid membrane: mechanisms of ion transport and toxic action.

The selectivity of ion channels produced by latrotoxin obtained from a black widow spider venom and by venom from the spider Steatoda paykulliana in bilayer phospholipid membrane was studied. Experimental current-voltage curves of these channels were used for the estimation of parameters of a two barrier model of their energy profiles. Selectivities of both types of channels are similar. Alkaline earth cations are permeable, the permeability increasing in the order Mg2+ less than Ca2+ less than Sr2+ less than Ba2+. In contrast transition metal cations block the channel, their efficiency decreases in the order: Cd2+ greater than or equal to Ni2+ greater than Zn2+ greater than Co2+ greater than Mn2+ (Steatoda paykulliana spider venom) and Cd2+ greater than Co2+ greater than Ni2+ greater than Zn2+ greater than Mn2+ (latrotoxin). Amplitudes of current carried by corresponding ions are mainly determined by the depth of the potential well for this ion, i.e., by its affinity to the cation binding site in the channel. The channels are also permeable to monovalent cations but they do not bind them. Selectivity for monovalent cations depends on Ca2+ concentration at the cis-side of membrane in the micromolar range. However, the addition of Ca2+ to the trans-side up to 10 mM does not affect currents carried by monovalent ions. It is suggested that venom-induced calcium channels have two conformational states with different selectivities which interconvert upon binding one calcium ion. Possible general schemes for the organisation of calcium channels in excitable membranes are also discussed. Finally, using a mathematical model of synaptic transmission, possible mechanisms of toxic action of spider venoms are considered.

Animals↗

Mygalomorph spider bites: a report on 91 cases in the state of São Paulo, Brazil.

From 1966 to 1991 91 cases of bites due to mygalomorph spiders were recorded at the Hospital Vital Brazil, Instituto Butantan, São Paulo, Brazil, representing less than 1% of all spider bites. The diagnosis was confirmed by positive identification of the spider involved. Envenoming is generally mild, the main symptom is local pain, and there is minor oedema and erythema. The data confirm the assumption that these nonaggressive spiders pose no health problem.

Adult↗

Brown spider dermonecrotic toxin directly induces nephrotoxicity.

Brown spider (Loxosceles genus) venom can induce dermonecrotic lesions at the bite site and systemic manifestations including fever, vomiting, convulsions, disseminated intravascular coagulation, hemolytic anemia and acute renal failure. The venom is composed of a mixture of proteins with several molecules biochemically and biologically well characterized. The mechanism by which the venom induces renal damage is unknown. By using mice exposed to Loxosceles intermedia recombinant dermonecrotic toxin (LiRecDT), we showed direct induction of renal injuries. Microscopic analysis of renal biopsies from dermonecrotic toxin-treated mice showed histological alterations including glomerular edema and tubular necrosis. Hyalinization of tubules with deposition of proteinaceous material in the tubule lumen, tubule epithelial cell vacuoles, tubular edema and epithelial cell lysis was also observed. Leukocytic infiltration was neither observed in the glomerulus nor the tubules. Renal vessels showed no sign of inflammatory response. Additionally, biochemical analyses showed such toxin-induced changes in renal function as urine alkalinization, hematuria and azotemia with elevation of blood urea nitrogen levels. Immunofluorescence with dermonecrotic toxin antibodies and confocal microscopy analysis showed deposition and direct binding of this toxin to renal intrinsic structures. By immunoblotting with a hyperimmune dermonecrotic toxin antiserum on renal lysates from toxin-treated mice, we detected a positive signal at the region of 33-35 kDa, which strengthens the idea that renal failure is directly induced by dermonecrotic toxin. Immunofluorescence reaction with dermonecrotic toxin antibodies revealed deposition and binding of this toxin directly in MDCK epithelial cells in culture. Similarly, dermonecrotic toxin treatment caused morphological alterations of MDCK cells including cytoplasmic vacuoles, blebs, evoked impaired spreading and detached cells from each other and from culture substratum. In addition, dermonecrotic toxin treatment of MDCK cells changed their viability evaluated by XTT and Neutral-Red Uptake methodologies. The present results point to brown spider dermonecrotic toxin cytotoxicity upon renal structures in vivo and renal cells in vitro and provide experimental evidence that this brown spider toxin is directly involved in nephrotoxicity evoked during Loxosceles spider venom accidents.

Amino Acid Sequence↗

An overview of peptide toxins from the venom of the Chinese bird spider Selenocosmia huwena Wang [=Ornithoctonus huwena (Wang)].

The bird spider Selenocosmia huwena Wang [=Ornithoctonus huwena (Wang)] is one of the most venomous spiders in China. The venom of this spider contains a mixture of compounds with different types of biological activity. About 400 proteins and peptides from the venom can be separated and detected by 2D electrophoresis. Of these, 14 peptide toxins have been purified and characterized from the venom of this spider, with several peptide toxins exhibiting structural similarity but high functional diversity. Most of these huwentoxins (HWTX) contain 30-40 amino acids with three disulfide bonds and adopt an "inhibitor cystine-knot" (ICK) motif in their three dimensional structure, except for huwentoxin-II (HWTX-II) which adopts a novel scaffold different from the ICK motif. As a group, the toxins possess quite different biological activities including inhibition of voltage-gated calcium and sodium channels, insecticidal activity, lectin-like agglutination, and inhibition of trypsin. Eight cDNAs encoding seven toxins, HWTX-I, -II, -III, -IIIa, -IV -V, and, -VII and one lectin, S. huwena lectin-I (SHL-I), have been cloned and sequenced. Comparison of the cDNA sequences of the eight peptides from S. huwena indicates that they can be classified into two different superfamilies according to the "prepro" region of their cDNA sequences.

Agglutination↗

Isolation and chemical characterization of PwTx-II: a novel alkaloid toxin from the venom of the spider Parawixia bistriata (Araneidae, Araneae).

Brazil has many species of spiders belonging to Araneidae family however, very little is known about the composition, chemical structure and mechanisms of action of the main venom components of these spiders. The main objective of this work was to isolate and to perform the chemical characterization of a novel beta-carboline toxin from the venom of the spider Parawixia bistriata, a typical species of the Brazilian 'cerrado'. The toxin was purified by RP-HPLC and structurally elucidated by using a combination of different spectroscopic techniques (UV, ESI-MS/MS and 1H NMR), which permitted the assignment of the molecular structure of a novel spider venom toxin, identified as 1-4-guanidinobutoxy-6-hydroxy-1,2,3,4-tetrahydro-beta-carboline, and referred to here as PwTx-II. This compound is toxic to insects (LD50 = 12+/-3 etag/mg honeybee), neurotoxic, convulsive and lethal to rats (LD50 = 9.75 mg/kg of male Wistar rat).

Animals↗

Insect-selective spider toxins targeting voltage-gated sodium channels.

The voltage-gated sodium (Na(v)) channel is a target for a number of drugs, insecticides and neurotoxins. These bind to at least seven identified neurotoxin binding sites and either block conductance or modulate Na(v) channel gating. A number of peptide neurotoxins from the venoms of araneomorph and mygalomorph spiders have been isolated and characterized and determined to interact with several of these sites. These all conform to an 'inhibitor cystine-knot' motif with structural, but not sequence homology, to a variety of other spider and marine snail toxins. Of these, spider toxins several show phyla-specificity and are being considered as lead compounds for the development of biopesticides. Hainantoxin-I appears to target site-1 to block Na(v) channel conductance. Magi 2 and Tx4(6-1) slow Na(v) channel inactivation via an interaction with site-3. The delta-palutoxins, and most likely mu-agatoxins and curtatoxins, target site-4. However, their action is complex with the mu-agatoxins causing a hyperpolarizing shift in the voltage-dependence of activation, an action analogous to scorpion beta-toxins, but with both delta-palutoxins and mu-agatoxins slowing Na(v) channel inactivation, a site-3-like action. In addition, several other spider neurotoxins, such as delta-atracotoxins, are known to target both insect and vertebrate Na(v) channels most likely as a result of the conserved structures within domains of voltage-gated ion channels across phyla. These toxins may provide tools to establish the molecular determinants of invertebrate selectivity. These studies are being greatly assisted by the determination of the pharmacophore of these toxins, but without precise identification of their binding site and mode of action their potential in the above areas remains underdeveloped.

Amino Acid Sequence↗

Acute conjunctival inflammation following contact with squashed spider contents.

PURPOSE: To report the effects to the eye following contact with spider contents. DESIGN: Observational case report. METHODS: A 46-year-old man presented with a painful and inflamed eye after contents of a spider got into his right eye when he squashed the spider with a newspaper. He suffered immediate severe pain in the eye, which rapidly became red and swollen. He had conjunctival edema and periorbital swelling of the eye and severe photophobia, but his vision was normal. RESULTS: Topical local anesthetic was applied to the right eye, which was then flushed with normal saline for an hour. The redness and swelling began to resolve over 2 hours. CONCLUSIONS: Ocular exposure to the contents of a spider can cause severe pain, swelling, and redness associated with photophobia. The effects are short term, so reassurance, irrigation, and symptomatic treatment are sufficient.

Acute Disease↗

The main products of the low molecular mass fraction in the venom of the spider Latrodectus menavodi.

The low molecular mass fraction of the venom of the spider Latrodectus menavodi, a black widow spider domiciled in Madagascar, was analyzed with HPLC combined with electrospray mass spectrometry as lyophilized powder and as freshly milked liquid venom. The main components were found to be the purine derivatives adenosine (1), guanosine (2), inosine (3), and 2,4,6-trihydroxypurine (4). Compounds 1-3 are known as components of spider or snake venoms, but they have never been found previously in the venom of the species Latrodectus, whereas compound 4 has not previously been found in any spider venoms. The lyophilized and unlyophilized venoms were found to have identical composition.

Adenosine↗

The venom optimisation hypothesis: a spider injects large venom quantities only into difficult prey types.

The spider Cupiennius salei needs 0.01-10 microl venom to kill a prey item. Since its venom glands contain only 10 microl and regeneration requires 8-16 days C. salei should use its venom very economically. By a monoclonal antibody we measured, for the first time, the amounts of venom injected by a spider into different prey types. Crickets and stick insects, as victims without special defence mechanism, received only the minimum amount of venom which is not significantly different from the LD(50). Blowflies and ground beetles received considerably more venom because they are difficult to overwhelm or even endanger the spider by their defence behaviour. These results support our venom optimisation hypothesis which supposes that spiders use their venom as economically as possible.

Animals↗

Protection against dermonecrotic and lethal activities of Loxosceles intermedia spider venom by immunization with a fused recombinant protein.

We report the use of a recombinant Loxosceles intermedia spider protein in the form of a fusion protein as an antigen for immunization in rabbits and mice. The aim is to produce model protective antisera in these animals against dermonecrotic and lethal activities of the venom from the Brazilian spider responsible for 3000 cases, reported annually, of spider bites in South Brazil. A protein homologous to the dermonecrotic toxin was cloned from a cDNA expression library made with L. intermedia venom glands, expressed in E. coli cells as a fusion protein with beta-galactosidase and the recombinant protein (Li-rec protein) was purified by molecular filtration and affinity chromatography [Kalapothakis et al., Toxicon (2002) in press]. The Li-rec protein was characterized and used as an antigen to generate antibodies in rabbits and mice. These specifically raised antibodies recognized the native venom. In vitro neutralization assay of lethal effects indicated that 1 ml of rabbit serum raised against Li-rec protein was able to neutralize 25 LD(50) of the whole venom. In vivo protection experiments, the fusion proteins induced a long-term protection in rabbits against the dermonecrotic activity of the native venom. Immunized mice were challenged with various doses of the Loxosceles venom. Mice were fully protected against 2.5 LD(50) of venom. This result provides basic data for the use of such recombinant spider proteins as immunogens in the development of anti-venoms for clinical use or can be used as a vaccine providing efficient immune protection against L. intermedia venom.

Animals↗

Envenomation by a spider, Agelenopsis aperta (family: Agelenidae) previously considered harmless.

This is the first report of clinically significant bites by the grass spider (Agelenopsis aperta). Two cases of envenomation in southern California are presented. In the more serious case, a child exhibited several envenomation symptoms. Although this spider may be generally innocuous, it should be considered as a creature of occasional medical importance. Many skin lesions are attributed to spider bites despite insufficient incriminating evidence, and bites that are clinically significant may often erroneously be attributed to the brown recluse spider. Misidentification can lead to undue anxiety in a patient's family.

Animals↗

Comparisons of the neurotoxic activity of the venom of several species of funnel web spiders (Atrax).

A bioassay has been developed to allow comparison of the neurotoxic activity of the venom of individuals of the four most common species of funnel web spiders (Atrax species). In the absence of purified Atrax neurotoxin, the stimulatory action of the venom on skeletal muscle from the cane toad, Bufo marinus, was compared with that of known concentrations of acetylcholine. Several experiments to validate the assay were performed, then venom samples from both male and female spiders were assayed. It was found that individual female spiders of all Atrax species normally secreted several times as much venom as male spiders, but its neurotoxicity was extremely variable and generally much lower. Both sexes tended to secrete more potent venom in summer than in other seasons. Both also showed some decrease in venom volumes and potencies if repeatedly milked at short intervals. All four Atrax species were found to have approximately equal levels of neurotoxic activity.

Acetylcholine↗

The brown spider Loxosceles laeta: source of the remedy Tarentula cubensis?

The homeopathic remedy Tarentula cubensis (Cuban tarantula), used in homeopathy to treat abscesses with burning pains, gangrene, septicaemia, toxaemia, has been grouped by homeopathic authorities with either the mygalomorph or wolf spiders. The original specimen used for preparation of the mother tincture was decomposed, leaving the spider's exact identity in doubt. Investigation of the toxicological and clinical literature, compared with homeopathic materia medica, reveals the brown spider, Loxosceles laeta, indigenous to South America but present also in Mid- and North America, as a more likely source. Venoms of spiders of the genus Loxosceles cause severe necrotic arachnidism, as well as, in some cases, a life-threatening systemic reaction marked by renal failure, disseminated intravascular coagulation, thrombocytopeania, coma and convulsions.

Animals↗

Lycotoxins, antimicrobial peptides from venom of the wolf spider Lycosa carolinensis.

Two peptide toxins with antimicrobial activity, lycotoxins I and II, were identified from venom of the wolf spider Lycosa carolinensis (Araneae: Lycosidae) by virtue of their abilities to reduce ion and voltage gradients across membranes. Both peptides were purified to homogeneity by reversed-phase liquid chromatography and determined to have the following primary structures by Edman microsequencing: IWLTALKFLGKHAAKHLAKQQLSKL-NH2 for lycotoxin I and KIKWFKTMKSIAKFIAKEQMKKHLGGE-OH for lycotoxin II. The predicted secondary structures of the lycotoxins display amphipathic alpha-helix character typical of antimicrobial pore-forming peptides. Antimicrobial assays showed that both lycotoxins potently inhibit the growth of bacteria (Escherichia coli) and yeast (Candida glabrata) at micromolar concentrations. To verify its hypothesized pore-forming activity, lycotoxin I was synthesized and shown to promote efflux of Ca2+ from synaptosomes, to cause hemolysis of erythrocytes, and to dissipate voltage gradients across muscle membrane. The lycotoxins may play a dual role in spider-prey interaction, functioning both in the prey capture strategy as well as to protect the spider from potentially infectious organisms arising from prey ingestion. Spider venoms may represent a potentially new source of novel antimicrobial agents with important medical implications.

Amino Acid Sequence↗

Jingzhaotoxin-III, a novel spider toxin inhibiting activation of voltage-gated sodium channel in rat cardiac myocytes.

We have isolated a cardiotoxin, denoted jingzhaotoxin-III (JZTX-III), from the venom of the Chinese spider Chilobrachys jingzhao. The toxin contains 36 residues stabilized by three intracellular disulfide bridges (I-IV, II-V, and III-VI), assigned by a chemical strategy of partial reduction and sequence analysis. Cloned and sequenced using 3'-rapid amplification of cDNA ends and 5'-rapid amplification of cDNA ends, the full-length cDNA encoded a 63-residue precursor of JZTX-III. Different from other spider peptides, it contains an uncommon endoproteolytic site (-X-Ser-) anterior to mature protein and the intervening regions of 5 residues, which is the smallest in spider toxin cDNAs identified to date. Under whole cell recording, JZTX-III showed no effects on voltage-gated sodium channels (VGSCs) or calcium channels in dorsal root ganglion neurons, whereas it significantly inhibited tetrodotoxin-resistant VGSCs with an IC(50) value of 0.38 microm in rat cardiac myocytes. Different from scorpion beta-toxins, it caused a 10-mV depolarizing shift in the channel activation threshold. The binding site for JZTX-III on VGSCs is further suggested to be site 4 with a simple competitive assay, which at 10 microm eliminated the slowing currents induced by Buthus martensi Karsch I (BMK-I, scorpion alpha-like toxin) completely. JZTX-III shows higher selectivity for VGSC isoforms than other spider toxins affecting VGSCs, and the toxin hopefully represents an important ligand for discriminating cardiac VGSC subtype.

Amino Acid Sequence↗