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[The chromosomal genes for black widow spider neurotoxins do not contain introns].

The overlapping fragments of the chromosomal DNA from black widow spider Latrodectus mactans carrying genes for high-molecular-mass protein neurotoxins, alpha- and delta-latroinsectotoxins (alpha-LIT and delta-LIT) and alpha-latrotoxin (alpha-LTX), were PCR-amplified and cloned. Restriction analysis of the PCR products showed that the distribution and sizes of the restriction fragments coincided with those deduced from the earlier sequencing of cDNAs of the corresponding genes. It thus followed that the alpha-LIT and delta-LIT genes are intronless. Along with our data on the structure of the alpha-latrocrustotoxin (alpha-LCT), this implies that the lack of introns is a common feature of the black widow spider genes encoding high molecular mass neurotoxins.

Animals↗

The effects of season of collection, feeding, maturation and gender on the potency of funnel-web spider (Atrax infensus) venom.

Assays of the venom of adult male and female as well as immature funnel-web spiders (Atrax infensus) were performed both at the end of the winter hibernation period and during the active summer months. For both sexes an increase in venom potency appeared to have occurred with the approach of summer, the venom of adult males then being clearly more potent than that of adult females. However, it was also found that, at least for adult females, forced abstinence from feeding was associated with an increase in venom potency, while feeding (at least on one insect species that A. infensus is known to accept as prey) seemed to lead to a lowered potency. From these observations it was concluded that the neurotoxin in the venom of female (and presumably also male) A. infensus spiders is probably synthesized in the venom glands rather than being extracted from the prey on which A. infensus normally feeds.

Animals↗

Purification of a neuroprotective component of Parawixia bistriata spider venom that enhances glutamate uptake.

(1) In this study, we examined the effects of crude venom from the spider Parawixia bistriata on glutamate and GABA uptake into synaptosomes prepared from rat cerebral cortex. Addition of venom to cortical synaptosomes stimulated glutamate uptake and inhibited GABA uptake in a concentration-dependent manner. (2) The venom was fractionated using reverse-phase high-performance liquid chromatography on a preparative column. The fraction that retained glutamate uptake-stimulating activity was further purified on a reverse-phase analytical column followed by ion-exchange chromatography. (3) The active fraction, referred to as PbTx1.2.3, stimulated glutamate uptake in synaptosomes without changing the K(M) value, and did not affect GABA uptake. Additional experiments showed that the enhancement of glutamate uptake by PbTx1.2.3 occurs when ionotropic glutamate receptors or voltage-gated sodium and calcium channels are completely inhibited or when GABA receptors and potassium channels are activated, indicating that the compound may have a direct action on the transporters. (4) In an experimental model for glaucoma in which rat retinas are subjected to ischemia followed by reperfusion, PbTx1.2.3 protected neurons from excitotoxic death in both outer and inner nuclear layers, and ganglion cell layers. (5) This active spider venom component may serve as a basis for designing therapeutic drugs that increase glutamate clearance and limit neurodegeneration.

Animals↗

Tetracycline protects against dermonecrosis induced by Loxosceles spider venom.

Envenomation by spiders belonging to the Loxosceles genus (brown spider) often results in local dermonecrotic lesions. We have previously shown that Loxosceles sphingomyelinase D (SMase D), the venom component responsible for all the pathological effects, induced the expression of matrix metalloproteinases (MMPs) in rabbits and in human keratinocytic cells. We also showed that the SMase D-induced apoptosis and MMP expression of keratinocytes was inhibited by tetracyclines. We have further investigated the ability of tetracyclines to inhibit or prevent the dermonecrotic lesion induced by Loxosceles venom in vivo and in vitro models. Primary cultures of rabbit fibroblasts incubated with increasing concentrations of venom or SMase D showed a decrease in cell viability, which was prevented by tetracyclines. In vivo experiments showed that topical treatments with tetracycline of rabbits, inoculated with crude Loxosceles intermedia venom or recombinant SMase D, significantly reduced the progression of the dermonecrotic lesion. Furthermore, tetracyclines also reduced the expression of MMP-2 and prevented the induction of MMP-9. Our results suggest that tetracycline may be an effective therapeutic agent for the treatment of cutaneous loxoscelism.

Animals↗

Brown recluse spider (Loxosceles reclusa) venom phospholipase D (PLD) generates lysophosphatidic acid (LPA).

Envenomation by the brown recluse spider (Loxosceles reclusa) may cause local dermonecrosis and, rarely, coagulopathies, kidney failure and death. A venom phospholipase, SMaseD (sphingomyelinase D), is responsible for the pathological manifestations of envenomation. Recently, the recombinant SMaseD from Loxosceles laeta was demonstrated to hydrolyse LPC (lysophosphatidylcholine) to produce LPA (lysophosphatidic acid) and choline. Therefore activation of LPA signalling pathways may be involved in some manifestations of Loxosceles envenomation. To begin investigating this idea, we cloned a full-length cDNA encoding L. reclusa SMaseD. The 305 amino acid sequence of the L. reclusa enzyme is 87, 85 and 60% identical with those of L. arizonica, L. intermedia and L. laeta respectively. The recombinant enzyme expressed in bacteria had broad substrate specificity. The lysophospholipids LPC, LPI (18:1-1-oleyol lysophosphatidylinositol), LPS, LPG (18:1-1-oleoyl-lysophosphatidylglycerol), LBPA (18:1-1-oleoyl-lysobisphosphatidic acid) (all with various acyl chains), lyso-platelet-activating factor (C16:0), cyclic phosphatidic acid and sphingomyelin were hydrolysed, whereas sphingosylphosphorylcholine, PC (phosphatidylcholine; C22:6, C20:4 and C6:0), oxidized PCs and PAF (platelet-activating factor; C16:0) were not hydrolysed. The PAF analogue, edelfosine, inhibited enzyme activity. Recombinant enzyme plus LPC (C18:1) induced the migration of A2058 melanoma cells, and this activity was blocked by the LPA receptor antagonist, VPC32183. The recombinant spider enzyme was haemolytic, but this activity was absent from catalytically inactive H37N (His37-->Asn) and H73N mutants. Our results demonstrate that Loxosceles phospholipase D hydrolyses a wider range of lysophospholipids than previously supposed, and thus the term 'SMaseD' is too limited in describing this enzyme.

Amino Acid Sequence↗

Lyme disease masquerading as brown recluse spider bite.

We report a case of Lyme disease with clinical features resembling those described from brown recluse spider bites. The most striking manifestation was a necrotic skin wound. Brown recluse spider bites may be overdiagnosed in some geographic regions. Tick bite and infection with Borrelia burgdorferi should be considered in the differential diagnosis of necrotic arachnidism in regions endemic for Lyme disease.

Animals↗

CSTX-13, a highly synergistically acting two-chain neurotoxic enhancer in the venom of the spider Cupiennius salei (Ctenidae).

The survival of the spider Cupiennius salei depends on its hunting success, which largely relies on its immediately paralyzing multicomponent venom. Here, we report on the isolation and characterization of CSTX-13, a neurotoxic enhancer in the spider venom. De novo elucidation of the disulfide bridge pattern of CSTX-13 and the neurotoxin CSTX-1 by tandem MS revealed an identical arrangement. However, in contrast to CSTX-1, CSTX-13 is a two-chain peptide with two interchain and two intrachain disulfide bridges. Furthermore, the insecticidal activity of CSTX-13 is synergistically increased in the presence of K+ ions as well as of the cytolytic peptide cupiennin 1a. We demonstrated that the weakly neurotoxic CSTX-13 enhances the paralytic activity of the neurotoxin CSTX-1 by 65% when it is administered with the latter at its entirely nontoxic physiological concentration, which is 440 times below its LD50 concentration.

Amino Acid Sequence↗

The cDNA and genomic DNA organization of a novel toxin SHT-I from spider Ornithoctonus huwena.

In present study the full-length cDNA of a novel toxin from the venom gland of spider Ornithoctonus huwena, named as SHT-I, has been cloned using the strategy of rapid amplification of cDNA ends, and then the whole genomic sequence of SHT-I (Selenocosmia huwena toxin-I) was determined using sequence-specific primers synthesized based on the acquired 3' and 5' ends of SHT-I cDNA sequence. It is unexpectedly found that intron was lacking in the genomic sequence of SHT-I. The result might evoke an interesting question whether the gene code of inhibitor cystine-knot peptides from spider venom is distinct from that of those known toxic peptides of scorpion and cone snail.

Amino Acid Sequence↗

Brown recluse spiders: a review to help guide physicians in nonendemic areas.

The brown recluse spider (Loxosceles reclusa) exists in significant populations mainly in the midwestern United States. Although bites can cause significant harm, envenomation is infrequent, and cases are usually clinically insignificant. Proper knowledge of the spider's habitat and lifestyle as well as the signs and symptoms of loxoscelism are needed to assess clinical cases adequately. Loxoscelism can masquerade as many serious pathologies, and vice versa, so it is important for the clinician to explore all possibilities in the differential diagnosis thoroughly. Treatments are controversial, and no conclusive test for envenomation is currently available. This review provides information to help physicians, especially in nonendemic areas, include or exclude brown recluse bite in the clinical diagnosis and provide proper care.

Animals↗

The structure of spider toxin huwentoxin-II with unique disulfide linkage: evidence for structural evolution.

The three-dimensional structure of huwentoxin-II (HWTX-II), an insecticidal peptide purified from the venom of spider Selenocosmia huwena with a unique disulfide bond linkage as I-III, II-V, and IV-VI, has been determined using 2D (1)H-NMR. The resulting structure of HWTX-II contains two beta-turns (C4-S7 and K24-W27) and a double-stranded antiparallel beta-sheet (W27-C29 and C34-K36). Although the C-terminal double-stranded beta-sheet cross-linked by two disulfide bonds (II-V and IV-VI in HWTX-II, II-V and III-VI in the ICK molecules) is conserved both in HWTX-II and the ICK molecules, the structure of HWTX-II is unexpected absence of the cystine knot because of its unique disulfide linkage. It suggests that HWTX-II adopts a novel scaffold different from the ICK motif that is adopted by all other spider toxin structures elucidated thus far. Furthermore, the structure of HWTX-II, which conforms to the disulfide-directed beta-hairpin (DDH) motif, not only supports the hypothesis that the ICK is a minor elaboration of the more ancestral DDH motif but also suggests that HWTX-II may have evolved from the same structural ancestor.

Amino Acid Sequence↗

Functional characterization of black widow spider neurotoxins synthesised in insect cells.

alpha-latrotoxin, alpha-latroinsectotoxin and the low-molecular-mass protein from black widow spider venom were synthesised in insect cells using the baculovirus expression system. SDS/PAGE analysis of recombinant-virus-infected cells revealed novel proteins that migrated with sizes similar to those of the neurotoxins from spider venom. The identities of these proteins as alpha-latrotoxin, alpha-latroinsectotoxin or the low-molecular-mass protein were confirmed by immunoblot analysis of infected cells with anti-(alpha-latrotoxin), anti-(alpha-latroinsectotoxin) or anti-(low-molecular-mass protein) IgG. Neither the low-molecular-mass protein nor alpha-latrotoxin were toxic upon injection into Trichoplusia ni larvae or upon virus-derived synthesis directly in the cytoplasm of the target tissue. Analysis of the biological activity of the recombinant virus encoding alpha-latroinsectotoxin, however, revealed a strong toxic effect on the T. ni larvae. These data indicate that the toxic effect of the native insectotoxin may be promoted by the alpha-latroinsectotoxin subunit alone and provides evidence that the mechanism of action of alpha-latroinsectotoxin may be mediated by internalisation of part of the neurotoxin alpha-subunit molecule.

Animals↗

Epidemiological and clinical manifestations of patients hospitalized with brown recluse spider bite.

BACKGROUND: Necrotic arachnidism represents a common health problem and standard treatments are usually safe and effective. OBJECTIVE: The aim of this study was to review documented Loxosceles species spider envenomations and identify the natural history of affected patients. METHODS: A retrospective single-centre study included 52 patients with necrotic arachnidism hospitalized in the dermatology department between 1997 and 2004. We examined the relationship between the epidemiological, clinical and laboratory parameters and degree of lesion severity, length of hospitalization and time to complete healing. RESULTS: The bites occurred predominantly in rural areas, 67% between April and August. Only 35% of the patients sought medical care within 24 h post bite. Most bites were of the extremities (67%). Time to complete healing ranged from 14 days to more than 8 weeks (mean, 4.8 weeks). A marked relationship was found between age, comorbidities, lesion severity and time to complete healing (P < 0.01). Duration of hospitalization was significantly longer in patients with severe thigh lesions (P < 0.02). CONCLUSIONS: Loxosceles species spider bites frequently induce necrotic, slowly healing ulcers on the fatty areas of the body. Early, appropriate systemic therapy may provide clinical benefit.

Adolescent↗

ADHESIVENESS OF SPIDER SILK.

Moths, by virtue of the loose scales that cover their wings and bodies, are admirably adapted to elude capture by orb-weaving spiders. Rather than sticking to the web, they may simply lose some of their scales to the viscid threads, and then fly on. Other insects, covered with detachable hairs or waxy powder, are similarly protected against entrapment. Quantitative data are presented on the adhesiveness of spider thread to insect cuticles oq various kinds.

Adhesiveness↗

Finding of primitive polyamine toxins in the venom of a joro spider, Nephila clavata.

A series of joro spider toxins, novel polyamines sharing a common moiety of 2,4-dihydroxyphenylacetyl cadaverine, have been identified using various bioassays, such as inhibition of a glutamatergic transmission and insecticidal activity. In this paper, we tried to chemically find still unknown polyamine toxins in the venom of a joro spider, N. clavata, by several analytical methods based on the characteristics of the common moiety. An aqueous extract from 3000 venoms was separated by preparative high performance liquid chromatography (HPLC). The polyamine toxins were detected by monitoring the fluorescence produced in an on-line reaction of o-phthalaldehyde with amino groups and UV absorption of the phenol group. Two compounds in minute quantity were purified and analyzed by gas-liquid chromatography (GC) and HPLC, which we specifically developed for the simultaneous determination of amino acids and polyamines of the toxins. Judging from the constituents of the hydrolysate by GC and HPLC and the molecular weights determined by fast atom bombardment mass spectrometry, the two compounds were estimated to be N-(2,4-dihydroxyphenylacetyl-L-asparaginyl)-N'- (3-aminopropyl-beta-alanyl) cadaverine and N-(4-hydroxyphenylacetyl-L-asparaginyl)-N'-(3-aminopropyl-beta-ala nyl) cadaverine. These compounds were small in content and molecular weight compared with hitherto known toxins. Both were presumed to be biochemically primitive toxins and were named spidamine and joramine, respectively.

Animals↗

Extracellular matrix molecules as targets for brown spider venom toxins.

Loxoscelism, the term used to describe lesions and clinical manifestations induced by brown spider's venom (Loxosceles genus), has attracted much attention over the last years. Brown spider bites have been reported to cause a local and acute inflammatory reaction that may evolve to dermonecrosis (a hallmark of envenomation) and hemorrhage at the bite site, besides systemic manifestations such as thrombocytopenia, disseminated intravascular coagulation, hemolysis, and renal failure. The molecular mechanisms by which Loxosceles venoms induce injury are currently under investigation. In this review, we focused on the latest reports describing the biological and physiopathological aspects of loxoscelism, with reference mainly to the proteases recently described as metalloproteases and serine proteases, as well as on the proteolytic effects triggered by L. intermedia venom upon extracellular matrix constituents such as fibronectin, fibrinogen, entactin and heparan sulfate proteoglycan, besides the disruptive activity of the venom on Engelbreth-Holm-Swarm basement membranes. Degradation of these extracellular matrix molecules and the observed disruption of basement membranes could be related to deleterious activities of the venom such as loss of vessel and glomerular integrity and spreading of the venom toxins to underlying tissues.

Animals↗

Funnel-web spider (Hadronyche infensa) envenomations in coastal south-east Queensland.

Five patients with confirmed funnel-web spider bites (Hadronyche infensa) presented to Nambour General Hospital, in south-east Queensland, between 1992 and 1998. Two patients required antivenom; low doses of antivenom were effective. Patients were bitten in spring and early summer. In areas such as this, where funnel-web spider bites are reported less frequently than in New South Wales, clinicians and the community should be aware of the risks and immediate management of these bites.

Antivenins↗

[Cloning and structure of gene encoded alpha-latrocrustoxin from the Black widow spider venom].

The primary structure of the crusta gene encoding alpha-latrocrustoxin (alpha-LCT), a high molecular mass neurotoxin specific to crustaceans, was determined in the black widow spider Latrodectus mactans tredicimguttatus genome. The total length of the sequenced DNA was 4693 bp. The structural part of the black widow spider chromosome gene encoding alpha-LCT does not contain introns. The sequenced DNA contains a single extended open reading frame (4185 bp) and encodes a protein precursor of alpha-LCT, comprising 1395 aa. We assume the Met residue at position -10 relative to the N-terminal residue of Glu1 of the mature toxin to be the first one in the protein precursor. The calculated molecular mass of the precursor (156147 Da) exceeds that of the mature toxin by approximately 30 kDa. These data are in agreement with the notion that over the course of maturation the protein precursor undergoes double processing--cleavage of a decapeptide from the N-terminal part and of a approximately 200-aa fragment from the C-terminal part. alpha-LCT displayed a number of imperfect ankyrin-like repeats and areas of structural homology with earlier studied latrotoxins; the highest homology degree (62%) was revealed with alpha-latroinsectotoxin (alpha-LIT).

Amino Acid Sequence↗

[The "hornet spider" of Plinius].

Pliny the Elder describes in his 'Natural History' XXIX 86 a species of spider dangerous for human beings and looking like a hornet without wings. This description corresponds to Solipugae of the genus Karschia, living in arid areas of Central Asia. These animals are not venomous but can cause harm by their powerful bite and in some cases by a following inflammation of the wound. On the contrary, an animal called "wasp-like", mentioned by Nikander of Kolophon ('Theriaca', v. 738-746) and often regarded to be the same creature, seems to be a typical venomous spider which causes an acute intoxication by its bite.

Animals↗