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Chromatographic detection of mushroom toxins other than Amanita phalloides in body fluids
53 2
Report and Abstracts
presence of hemolytic, musculotoxic and neurotoxic protein components that have a range of molecular weights from 10,000 to 600,000. Various techniques have been attempted in the purification process of these active components . In this report, the purification of the active and neurotoxic proteins was attempted using a preparative isoelectric focussing machine obtained from Bio-Rad (Rotofor) . Proteins obtained were characterised and analysed on SDS-PAGE and analytical iscelectric focussing. The use of this method is evaluated in comparison to others that have been previously employed (Othman et al., 1991) . In the second part of this report, the research group was invited to test out the potency of an antidote that was made traditionally using a combination of plant extracts . The effectiveness of these extracts for treatment ofjellyfish stings was explored in an experimental basis. This method was adopted due to the àparsity of human envenomation and evaluation can only be done on animals in the laboratory . Using a known am8ùnt (wet weight) of a mixture of extracts of various plants, the effect of Chironex intoxication was investigated ié the presence and absence of this plant mixture in mice . Preliminary results indicated that treatment with the plant extracts provided 80-100% protection against Chironex envenomation when injected intravenously with toxin extract. However, injection of the mixture 5 min after toxin injection did not show significant protection as observed with the injection of toxin and plant mixture together . Similar observation were also obtained when anti-venom was used instead of plant mixtures . It should also be noted that the protocol use in experiments were not in accordance with the prescribed use of this antidote, which specifies the treatment on externally affected areas only. OTHMAN, I . and Buxxerr, J . W . (1991) Toxicon 28, 821~35 . Cases ojjellyftsh envenomation in Malaysia and some cytotoxic studies of their toxins . N . Aztu and I . OTHMAN (Department of Biochemistry, Faculty of Medicine, University of Malaya, 59100 Kuala Lumpur, Malaysia) . Ttn: sEns around Malaysia contain numerous varieties of jellyfish . In the past, serious and lethal envenomation by jellyfish were rarely reported in Malaysia since most of the species found here are not toxic, and accidental encounters with these animals cause mild injury requiring only topical medication and out-patient treatment . The most serious injury has been associated with the species Carybdea rastoni, which is found in abundance around Penang island. However, in the last few years serious and lethal envenomation has been frequently reported, especially in the waters around Labuan and other islands North of Sabah . Fresh wounds and scar patterns seen on some of the patients indicated possible existence of multi-tentacled box jellyfish in these waters . Investigations are ongoing to determine whether the box jellyfish is of the species Chironexrfleckeri or Chiropsalrnus quadrigarus . The obvious immediate effect of envenomation by various jellyfish is the local reaction of the skin upon contact with the venom, which ranges from edema and urticaria to severe necrosis of the affected area . Edema and hemorrhagic effects had been shown with various species found in the Malaysian waters . The present in vitro studies on the effects of extract from Chironex Jleckeri and Carybdea rastoni on two cell lines, spleen hybridoma NAD(L) 3G410 and fibroblast from human epidermal carcinoma HEP-2, showed that they are cytotoxic . Cytotoxicity is manifested as changes in morphological appearance of the cells, loss of adherence and inhibition ofDNA synthesis. Crude extracts (CE) caused obvious morphological changes to these cells while acetone soluble extracts (AE) which did not cause obvious morphological changes, were more potent in inhibiting DNA synthesis . The observation that the extracts did not cause immediate lysis but caused gradual morphological changes over 24 hr indicated that they may cause apoptosis . Chromatographic detection of mushroom toxins other than Amanita phalloides in body fluids. D . Mtct~LOr (Laboratoire de Chimie du Muséum National d'Histoire Naturelle (URA 401 du CNRS), 63 rue Buffos, 75005 Paris, France) . MUSIfltOOM poisonings occur every year, in several cases they lead to deaths or failure of organs such as kidneys . Chromatographic methods for identification of higher fungi toxins have been developed in order, on the one hand, to detect the toxic agents, since the victim often does not associate his condition with a mushroom meal consumed several hours or days earlier, and, on the other hand, to monitor the appropriate treatment oriented towards elimination of the tonic agent . Our current knowledge of the different poisoning and of the various methods available for detection and quantitation of the toxins are reviewed here. Effects ojwaglerin-1, a toxin from Trimeresurus wagleri, on the neuromuscular transmission ojmouse ntrue-muscle preparations . W. H . HsrEx,~ M. C . TsN,' C . Y. Lam' and L . A . SunrttZ (~ Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan, R .O .C . ; and tU .S. Army Medical Research Institute of Infectious Disease, Fort Detrick, Frederick, MD 21702-5011, U .S .A .) . E~ecrs of waglerin-I, a toxin from Trimeresurus wagleri, on the neuromuscular (NM) transmission were studied on the mouse phrenic nerve~iaphragm preparation and Triangularis sterni nerve-muscle preparation. The toxin
Report and Abstracts
presence of hemolytic, musculotoxic and neurotoxic protein components that have a range of molecular weights from 10,000 to 600,000. Various techniques have been attempted in the purification process of these active components . In this report, the purification of the active and neurotoxic proteins was attempted using a preparative isoelectric focussing machine obtained from Bio-Rad (Rotofor) . Proteins obtained were characterised and analysed on SDS-PAGE and analytical iscelectric focussing. The use of this method is evaluated in comparison to others that have been previously employed (Othman et al., 1991) . In the second part of this report, the research group was invited to test out the potency of an antidote that was made traditionally using a combination of plant extracts . The effectiveness of these extracts for treatment ofjellyfish stings was explored in an experimental basis. This method was adopted due to the àparsity of human envenomation and evaluation can only be done on animals in the laboratory . Using a known am8ùnt (wet weight) of a mixture of extracts of various plants, the effect of Chironex intoxication was investigated ié the presence and absence of this plant mixture in mice . Preliminary results indicated that treatment with the plant extracts provided 80-100% protection against Chironex envenomation when injected intravenously with toxin extract. However, injection of the mixture 5 min after toxin injection did not show significant protection as observed with the injection of toxin and plant mixture together . Similar observation were also obtained when anti-venom was used instead of plant mixtures . It should also be noted that the protocol use in experiments were not in accordance with the prescribed use of this antidote, which specifies the treatment on externally affected areas only. OTHMAN, I . and Buxxerr, J . W . (1991) Toxicon 28, 821~35 . Cases ojjellyftsh envenomation in Malaysia and some cytotoxic studies of their toxins . N . Aztu and I . OTHMAN (Department of Biochemistry, Faculty of Medicine, University of Malaya, 59100 Kuala Lumpur, Malaysia) . Ttn: sEns around Malaysia contain numerous varieties of jellyfish . In the past, serious and lethal envenomation by jellyfish were rarely reported in Malaysia since most of the species found here are not toxic, and accidental encounters with these animals cause mild injury requiring only topical medication and out-patient treatment . The most serious injury has been associated with the species Carybdea rastoni, which is found in abundance around Penang island. However, in the last few years serious and lethal envenomation has been frequently reported, especially in the waters around Labuan and other islands North of Sabah . Fresh wounds and scar patterns seen on some of the patients indicated possible existence of multi-tentacled box jellyfish in these waters . Investigations are ongoing to determine whether the box jellyfish is of the species Chironexrfleckeri or Chiropsalrnus quadrigarus . The obvious immediate effect of envenomation by various jellyfish is the local reaction of the skin upon contact with the venom, which ranges from edema and urticaria to severe necrosis of the affected area . Edema and hemorrhagic effects had been shown with various species found in the Malaysian waters . The present in vitro studies on the effects of extract from Chironex Jleckeri and Carybdea rastoni on two cell lines, spleen hybridoma NAD(L) 3G410 and fibroblast from human epidermal carcinoma HEP-2, showed that they are cytotoxic . Cytotoxicity is manifested as changes in morphological appearance of the cells, loss of adherence and inhibition ofDNA synthesis. Crude extracts (CE) caused obvious morphological changes to these cells while acetone soluble extracts (AE) which did not cause obvious morphological changes, were more potent in inhibiting DNA synthesis . The observation that the extracts did not cause immediate lysis but caused gradual morphological changes over 24 hr indicated that they may cause apoptosis . Chromatographic detection of mushroom toxins other than Amanita phalloides in body fluids. D . Mtct~LOr (Laboratoire de Chimie du Muséum National d'Histoire Naturelle (URA 401 du CNRS), 63 rue Buffos, 75005 Paris, France) . MUSIfltOOM poisonings occur every year, in several cases they lead to deaths or failure of organs such as kidneys . Chromatographic methods for identification of higher fungi toxins have been developed in order, on the one hand, to detect the toxic agents, since the victim often does not associate his condition with a mushroom meal consumed several hours or days earlier, and, on the other hand, to monitor the appropriate treatment oriented towards elimination of the tonic agent . Our current knowledge of the different poisoning and of the various methods available for detection and quantitation of the toxins are reviewed here. Effects ojwaglerin-1, a toxin from Trimeresurus wagleri, on the neuromuscular transmission ojmouse ntrue-muscle preparations . W. H . HsrEx,~ M. C . TsN,' C . Y. Lam' and L . A . SunrttZ (~ Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan, R .O .C . ; and tU .S. Army Medical Research Institute of Infectious Disease, Fort Detrick, Frederick, MD 21702-5011, U .S .A .) . E~ecrs of waglerin-I, a toxin from Trimeresurus wagleri, on the neuromuscular (NM) transmission were studied on the mouse phrenic nerve~iaphragm preparation and Triangularis sterni nerve-muscle preparation. The toxin