[Jellyfish sting injuries]. / Durch Quallen verursachte Verletzungen.

BACKGROUND: Jellyfish are distributed worldwide; they cause local skin injuries upon contact which are often followed by systemic signs of envenoming. OBJECTIVES: Which jellyfish species are of medical importance, which skin reactions and systemic symptoms occur, which first-aid measures and treatment options exist? METHODS: Review of the medical literature and discussion of first-aid and therapeutic options. RESULTS: Jellyfish capable of causing skin injuries occur in almost all oceans. Several jellyfish species may cause severe, potentially lethal, systemic symptoms; they include the Portuguese man-of-war (Physalia physalis) and box jellyfish (Chironex fleckeri, Carukia barnesi, Chiropsalmus quadrigatus). CONCLUSIONS: Among the injuries and envenoming symptoms caused by marine organisms, jellyfish dermatitis should not be underestimated. Skin reactions may not only a dermatological problem, but also be accompanied by complex systemic toxic symptoms which are a challenge for internists.

[Severe encephalopathy after taking a "phytopharmacon" from Vietnam]. / Schwere Enzephalopathie nach Einnahme eines "Phytopharmakons" aus Vietnam.

HISTORY: A 59-year-old woman went into coma after she had taken a phytopharmacon from Vietnamese generally used as an antidiabetic drug to treat her skin disease. INVESTIGATIONS: CT-scans revealed signs of pneumonia and cerebral edema. Severe brain damage was diagnosed by MRI-investigation. By chemical-toxicological analysis of the drug glibenclamide was identified in a concentration of 1.1 mg/g. DIAGNOSIS, TREATMENT AND COURSE: The patient developed severe irreversible encephalopathy. By neurorehabilitative treatment her physical status slightly improved, but she died after 13 months without regaining consciousness. CONCLUSIONS: The cause of losing consciousness remained unclear, however, severe hypoglycaemia following the use of the drug may be taken into account. An urgent warning against the use of such preparations is highly recommended.

Puffer fish poisoning in Bangladesh: clinical and toxicological results from large outbreaks in 2008.

Poisoning after eating puffer fish containing highly lethal tetrodotoxin (TTX) is widespread in Asia. In 2008, naïve inland populations in Bangladesh were exposed to cheap puffer fish sold on markets. In three outbreaks, 141 patients with history of puffer fish consumption were hospitalized. Symptoms of poisoning included perioral paraesthesia, tingling over the entire body, nausea and vomiting, dizziness, headache, abdominal pain and muscular paralysis of the limbs. Seventeen patients (12%) died from rapidly developing respiratory arrest. Blood and urine samples from 38 patients were analyzed using a TTX-specific enzyme-linked immunoassay (ELISA). Medium to high TTX levels were detected (1.7-13.7 ng/ml) in the blood of 27 patients. TTX was below detection level (< 1.6 ng/ml) in 11 blood samples but the toxin was detected in urine. Ten patients had blood levels above 9 ng/ml and developed paralysis; seven of these died. The remaining patients recovered with supportive treatment. High concentrations of TTX and its analogues 4-epiTTX and 4,9-anhydroTTX were also found in cooked puffer fish by post-column liquid chromatography-fluorescence detection. To prevent future instances of puffer fish poisoning of this magnitude, measures should be implemented to increase awareness, to control markets and to establish toxicological testing. To improve the management of this and other poisoning in Bangladesh, facilities for life-saving assisted ventilation and related training of healthcare personnel are urgently needed at all levels of the health system.

New conopeptides of the D-superfamily selectively inhibiting neuronal nicotinic acetylcholine receptors.

The venom of cone snails (Conus spp.) is a rich source of peptides exhibiting a wide variety of biological activities. Several of these conopeptides are neuronal nicotinic acetylcholine receptor (nAChR) antagonists and belong to the A-, M-, S-, C and the recently described D-superfamily (alphaD-conopeptides). Here we describe the discovery and characterization of two alphaD-conopeptides isolated from the venom of Conus mustelinus and Conus capitaneus. Their primary structure was determined by Edman degradation, MS/MS analysis and by a PCR based approach. These peptides show close structural homology to the alphaD-VxXIIA, -B and -C conopeptides from the venom of Conus vexillum and are dimers (about 11kDa) of similar or identical peptides with 49 amino acid residues and a characteristic arrangement of ten conserved cysteine residues. These novel types of conopeptides specifically block neuronal nAChRs of the alpha7, alpha3beta2 and alpha4beta2 subtypes in nanomolar concentrations. Due to their high affinity, these new ligands may provide a tool to decipher the localisation and function of the various neuronal nAChRs.

[Snakes, scorpions and other poisonous creatures: prophylaxis and emergency medicine]. / Erste Hilfe bei Bissen und Stichen durch Gifttiere: Zwischenfall im Paradies.

Most cases of poisoning by animals are caused by snakes, spiders or scorpions. In addition to"first aid" such as calming the victim and immobilization of the bitten limb, other measures include monitoring vital functions and rapid transportation to hospital as dictated by the symptoms presenting, and, where indicated, injection of an antiserum (identification of the animal concerned). On no account should the bite wound be cut or attempts made to suck out the venom. While the sting of a scorpion or a spider bite often do not lead to severe complications in adults, a brush with a poisonous snake may be much more serious.

Primary structure and electrophysiological characterization of two almost identical isoforms of toxin from Isometrus vittatus (family: Buthidae) scorpion venom.

Two almost identical proteins with 70 amino acid residues each, closely packed by four disufide bridges, and molecular masses of 7899.5 and 7884.7 were isolated and sequenced from the venom of the scorpion Isometrus vittatus from Pakistan. They differ by an acidic amino acid residue (glutamic or aspartic) at the same position 55 of the peptide chain, however, they exhibit the same length, the same charge and are undistinguishable when separated by C(18) reverse phase HPLC. The mixture of the two proteins called IsomTx1 depolarizes the cockroach isolated axon; artificial repolarization is followed by sustained repetitive activity, artificial hyperpolarization facilitates bursting activity observed as an answer to rapid depolarization to -60 mV. The depolarization is antagonized by TTX. In voltage-clamp experiments IsomTx1 increases axonal sodium permeability which has a particular importance between resting and threshold potentials and moderately slows down the fast inactivation. These characteristics closely resemble those of other anti-insect scorpion toxins classified as contractive toxins from Androctonus and Buthotus venoms.

The levels of tetrodotoxin and its analogue 6-epitetrodotoxin in the red-spotted newt, Notophthalmus viridescens.

Tetrodotoxin (TTX) and its analogue 6-epiTTX were detected in 11-12 specimens of the red-spotted newt, Notophthalmus viridescens, by a post-column fluorescent-HPLC system and by LC/MS in selected ion monitoring mode. TTX levels varied considerably among individuals from low (less than 0.15 microg TTX/g newt) to high concentrations (23.5 microg TTX/g newt), while 6-epiTTX was found to be a minor constituent in all specimens.

Toxicity in animals. Trends in evolution?

Animals acquire toxicity either by metabolic synthesis of toxins (secondary metabolites), by expression of toxin genes or by the uptake, storage and sequestration of toxins produced by other organisms, i.e., microbes, plants or other animals. Variability of toxin structure and function is high. Peptide toxins in particular, although relying on a limited number of structural frameworks, often exhibit considerable structural hypervariability. An accelerated rate of evolution in the toxin gene structure (conserved introns, but high substitution rates in the exons) leads to the functional diversity of these peptides or proteins. The selective forces which may drive toxin evolution are unknown. Venomousness or the possession of toxins can be essential for survival, but the advantage of toxin biosynthesis may also be of minor importance or has been lost during evolution.

The antihemorrhagic factor, erinacin, from the European hedgehog (Erinaceus europaeus), a metalloprotease inhibitor of large molecular size possessing ficolin/opsonin P35 lectin domains.

From muscle extracts of the European hedgehog, Erinaceus europaeus, an antihemorrhagic factor, erinacin, was purified by ammonium sulfate precipitation followed by chromatography on DEAE-cellulose, hydroxylapatite and gel filtration columns. A purification of approx. 1400-fold was achieved with an overall yield of 21% in antihemorrhagic activity. The molecular weight of erinacin determined by gel filtration was approx. 1000 kDa. SDS-PAGE of erinacin under reducing conditions indicates that it consists of two types of subunits, alpha and beta, with molecular weights of 37 and 35 kDa, respectively, in a ratio of 1:2. In the presence of 6 M guanidine-HCl, erinacin dissociates into alpha-subunits and beta-subunit decamers. From these results the subunit assembling of erinacin has been formulated as alpha(10).2beta(10). The molecular weight of the subunits and of the beta-subunit decamer was confirmed by MALDI-TOF mass spectrometry. Erinacin inhibits the hemorrhagic and proteolytic activity of the major hemorrhagic metalloprotease from the venom of Bothrops jararaca. Complete inhibition was achieved in an equimolar mixture of inhibitor and enzyme suggesting an equimolar complex. Erinacin is not inhibiting serine proteases such as trypsin and chymotrypsin, it was characterized to be a metalloprotease inhibitor. In electronmicroscopy, flower bouquet-like structures characteristic for some animal lectins were observed. Amino acid sequence analysis indicated that both subunits are almost identical and are composed of common amino terminal, collagen- and fibrinogen-like domains homologous to proteins of the ficolin/opsonin P35 lectin family.

Amino acid sequence studies on cytolytic toxins from sea anemone Heteractis magnifica, Entacmaea quadricolor and Stichodactyla mertensii (Anthozoa).

The complete amino acid sequence of a cytolytic toxin, HmT, isolated from sea anemone Heteractis magnifica was determined. It is composed of 177 amino acid residues and lacks half-cystines. Partial N-terminal sequences of three other cytolysins from Entacmaea quadricolor (EnT) and Stichodactyla mertensii (SmT-1 and SmT-2) were also determined. Comparing these sequences with those of other sea anemone cytolysins, a high degree of homology was observed.

Notes on the traditional use of plants to treat snake bite in northern Papua New Guinea.

Information on snake bite treatment using plants was collected in three areas of northern Papua New Guinea. Liquid from six plants is either placed topically on the bite site or plant parts (bark) are chewed and the sap is swallowed. However, beside tannins in the bark no secondary metabolites, e.g. alkaloids, steroids, saponins or flavonoids have been detected by thin-layer chromatography in alcoholic extracts of the plants involved rendering this type of snake bite treatment questionable.

Distribution and sequestration of palytoxin in coral reef animals.

In the reefs off the Colombian coast (Caribbean Sea) and around Lizard Island, Australia (Pacific), palytoxin (PTX), which has been detected in zoanthid species of the genus Palythoa, also occurred in various other marine organisms living in close association with zoanthid colonies, e.g. sponges (Porifera), soft corals (Alcyonaria), gorgonians (Gorgonaria), mussels, and crustaceans. Predators, e.g. polychaete worms (Hermodice carunculata), a starfish (Acanthaster planci) and fish (Chaetodon species) feeding on Palythoa colonies, accumulate high toxin concentrations in their organs, where PTX is stored in its active form. The high level of toxin tolerance observed in marine animals may enable the wide distribution of PTX in marine biota and its transport and sequestration in food chains.

Studies on biological and enzymatic activities of salivary glands from the European hedgehog (Erinaceus europaeus).

Aqueous extracts of salivary glands (Glandula submandibularis and G. parotis) from the European hedgehog (Erinaceus europaeus) exhibited neither lethal effect (intraperitoneal injection, mice), nor haemorrhagic and myonecrotic (mice) activity. Of the various enzymes tested (kallikrein, casein hydrolysis, phospholipase A2, acid and alkaline phosphatase, alpha-amylase), both glands possessed alkaline phosphatase and alpha-amylase activity only. These experiments suggest that toxic saliva in mammals is restricted to certain insectivores (shrews and solenodons) only.

Effects of a centipede venom fraction on insect nervous system, a native Xenopus oocyte receptor and on an expressed Drosophila muscarinic receptor.

Centipede venoms are complex protein mixtures; very few is known about their pharmacological actions. Application of a Scolopendra sp. venom fraction (SC1) on the cockroach giant axon induced an increase in the leak current correlated with a decrease in the membrane resistance, suggesting the presence in SC1 of components opening non-specific pores in the axonal membrane. On a cockroach central cholinergic synapse, microinjection of SC1 induced a small transient depolarization of the postsynaptic membrane, followed by a slow stable depolarization and a drastic decrease in the evoked subthreshold excitatory postsynaptic potential amplitude. A pretreatment of the ganglion with atropine or scopolamine reduced the amplitude of the SC1-induced depolarizing wave, suggesting a possible cholinergic muscarinic target. On control Xenopus oocytes, SC1 induced an inward oscillatory Ca2(+)-dependent Cl- current mediated through the activation of native lysophosphatidic acid receptors (LPAr). Indeed, pretreatment of oocytes with 1 microM N-palmitoyl-tyrosine phosphoric acid, a selective competitive antagonist of LPAr, decreased responses to SC1 by 70%. Application of SC1 to oocytes expressing a cloned Drosophila muscarinic receptor (Dml) induced a biphasic response comprising: (1) a large fast Cl- current that was abolished by pretreatment with atropine and scopolamine and (2) a slow and small oscillating Cl- current corresponding to the response observed in control oocytes. These observations confirm the presence of muscarinic agonists in SCI and reveal their direct action on an insect muscarinic receptor subtype homologous to mammalian M1-M3 receptors.

RFLP and sequence analysis of the cytochrome b gene of selected animals and man: methodology and forensic application.

To identify common animal species by analysis of the cytochrome b gene a method has been developed to obtain PCR products of a large domain of the cytochrome b gene (981 bp out of 1140 bp) in humans, selected mammals and birds using the same specifically designed primers. Species-specific RFLP patterns are generated by co-restriction with the restriction endonucleases ALU I and NCO I. The RFLP patterns obtained are conclusive even in mixtures of two or more species. The results were confirmed by sequence analysis which in addition explained intraspecies variations in the RFLP patterns. The method has been applied to forensic casework studies where the origin of roasted meat, stomach contents and a bone sample has been successfully identified.

Occurrence and sequestration of toxins in food chains.

Animals may acquire toxicity by absorbing toxic compounds from their food, e.g. from plants or other animals. Sequestration and accumulation of toxins may provide protection from predators, which learn to avoid this prey because of unpleasant experiences such as bitter taste. This is a common phenomenon in marine as well as in terrestrial ecosystems. Moreover, toxins may enter food chains where they accumulate reaching high, often lethal concentrations. Palytoxin which had been primarily detected in marine zoanthids (Palythoa sp.), occurs also in a wide range of other animals, e.g. in sponges, corals, shellfish, polychaetes and crustaceans, but also in fish, which feed on crustaceans and zoanthids as well. These animals exhibit a high resistance to the toxin's action. The mechanisms which protect the Na+, K+-ATPase of their cell membranes, the primary target of palytoxin, is unknown. Sequestration of the toxin by other animals may cause health problems due to food poisoning.