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Biochemical assay using the aqueous extract of Ptychodera flava (hemichordate) on the respiratory enzyme phenoloxidase
First A s i a - Pacific Congress
15
ABSTRACTS The abstracts are arranged alphabetically by first author. Only abstracts of work which was actually presented at the meeting are included in the following list.
Experimental studies on the enteropathogenicity ofVibrio fluvialis. R. K. AOARWALand S. C. SAr~YAL(Enteric Infection Unit, Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi221005, India). STRAINS of Vibriofluvialis isolated from diverse sources were tested for enterotoxigenicity in rabbit ileal loops, rabbit's skin and CHO cells and for enteroinvasiveness in rabbit's eye. Live cells and culture filtrates of all the strains of V. fluvialis tested were found to be enterotoxigenic. The enterotoxic factor was found to be heat labile, as evidenced by heat treatment, time course of fluid accumulation and negative suckling mouse assay. The outpouring of fluid into the loops was inhibited by chlorpromazine, indicating that the enterotoxin may act through mediation of cAMP. The tox gent(s) appears to be chromosomal, as the cells could not be cured of enterotoxicity by different methods used for elimination of plasmids. None of the strains tested could produce keratoconjunctivitis in rabbit's eye, indicating lack of invasive capability.
Clinico-pathological correlation in Russell's viper bite. MAUNOMAUNOAYE (Department of Surgery, Changi Hospital, Singapore). THIS presentation is based on personal clinical experience of 905 snake bite patients treated at Insein Hospital, Rangoon, Burma, over a ten year period. Among 905 patients, 434 victims were bitten by Russell's viper, which caused 145 deaths; post-mortem examination was performed on every fatal case. Russell's viper bite inflicts local reactions such as fang marks, oedema, blister, necrosis and even gangrene. More importantly, severe envenomation produces systemic manifestation such as shock, blood coagulation defect, spontaneous haemorrhages, acute renal failure, pituitary necrosis and even death. Cfinical features of Russell's viper bite will be illustrated and correlated with morbid and microscopic pathology. An outline of management of viper envenomation will also be presented.
Toxicology o f the plant Calotropis gigantea. J. AZARIAH, H. AT-~IAH, S. MALLIKESAN,M. V. SUMATHIand C. StmDARA~J (Department of Zoology and Centre for Advanced Study in Botany, University of Madras, Guindy Campus, Madras 600 025, India). IN TAMILNADUthe plant Calotropisgigantea is harvested and used for agricultural practices, especially in paddy cultivation. The plant is also used to cause abortion and homicide. Hence, a detailed literature survey and a few laboratory experiments were made. Information on the traditional medical use (non-scientific) and scientific work on the toxic nature of the plants was gathered. Paddy variety ADT 36, fresh water mussel Lamellidens marginalis and the mitotic index of root tips of Ailium cepa were used as biological and genetic screeing material to assess the toxicity of the plant extract. Aqueous extracts of the plant were made and other dilutions were made as required. It was found that the extract was toxic in high concentrations and with progressive dilutions seed germination was stimulated. Similarly, the extract was toxic to the survival of the mussels at high concentrations. The heart beat and the ciliary beat of the gill were seen to slow down at high concentrations. Lower dilutions accelerated both the functions. However, it was found that root germination of onion was inhibited due to cytotoxicity. There were both nuclear and chromosomal aberrations. Since people use the plants to cause abortion and in other treatments, it is recommended that the public be educated on its ill effects. A need to standardise the traditional use is suggested.
Biochemical assay using the aqueous extract o f Ptychodera flava (hemichordate) on the respiratory enzyme phenoloxiduse. P. S. R. BAI and J. AZARIAH(Department of Zoology, University of Madras, Guindy Campus, Madras 600 025, India). IT HAS BEEN established that the aqueous extract of Ptychodera flava can inhibit or stimulate the clevage of fertilized eggs of marine polychaetes. A study was made to assess the presence of physiologically active organic compounds. The effect of the extract was studied on the respiratory enzyme phenoloxidase of the surf crab Emerita emeritus. The extract was prepared on the recommendations outlined in a previous publication. On the
16
First A s i a - Pacific Congress
basis of the enzyme activity, the physiological activity of the extract was assayed at 480 nm using a spectrophotometer. The assay revealed that enzyme activity decreased soon after administration of the extract. The results indicated that there are pharmacologically and physiologically active compounds which can inhibit the enzyme activity depending upon its storage period, percentage composition and dosage adminstered. The Indian P. tiara has been characterized as containing iodocresol and ethylphenol compounds. There is no trace of indoles and hydroquinones. The phenol was found to be linked with bromine and iodine, whereas P. flava of western waters contained more bromophenols. It is concluded that the halometabolites may be responsible for the observed changes in enzyme activity.
Clinical features on 19,890 cases o f ciguatera (fish poisoning) in French Polynesia. R. A. BAGNIS (Institut Territorial de Recherces M6dicales Louis Malard6, BP 30, Papeete, Tahiti, Polyn6sie Fran~aise). THE EVALUATION of patients took place as part of an epidemiological program from 1964 to 1986. A standardized l-page questionnaire was used to obtain most of the clinical data. The information was gathered from patients in Society, Marquesas, Tuamotu, Gambier and the Austral Islands, in which ciguatera is known to be a relatively common problem. The majority of the questionnaire were completed by an examining physician or nurse. The most common symptoms could be grouped into four main categories: neurological, digestive, cardiovascular and generalized. From these, paraesthesia, arthralgia, myalgia, vertigo, ataxia, diarrhea, vomiting, nausea, bradycardia, hypotension, pruritus and asthenia account for the most frequent or most typical, with a frequency ranging from 90 to 15%. Approximately 77°70 of the patients became symptomatic less than 12 h after ingestion of the fish. The recovery period may vary from 72 h to several days, weeks or months. The rate of fatality was less than 1% 0, death generally occuring more from cardiac than from respiratory failure. Significant differences in certain symptoms and signs, as well as in evolutionary pattern, may be noted according to species of fish, susceptibility of patients and recurrence of illness. Being poisoned multiple times may result in a clinically more severe illness than disease observed in patients experiencing ciguatera for the first time. Hypersensitization to fish or seafood may be developed by some patients, resulting in allergic-like cutaneous, respiratory or digestive features.
Suppression o f oxidative phosphorylation in mitochondria by hornet venom. L. BARR-NEA,D. SHAHARand J. ISHAY (Suckler School of Medicine, Tel Aviv University, Ramat Aviv, Israel). THE PURPOSE of this work was to estimate the direct influence of hornet (Vespa orientalis) venom on liver mitochondria in vitro, since in vivo morphological studies had shown that the number of mitochondria and amount of their cristae was decreased in the liver of rats treated with hornet venom. The liver was homogenized and a mitochondrial preparation obtained after centrifugation at 10,000 g. The mitochondria were then incubated in different substrates for estimation of the activity of the different sites of electron transfer: malate or pyruvate for site I; succinate for site II; ascorbate for site III. Oxygen consumption was also estimated in an oxygen electrode. The results were expressed as nl oxygen atom/min/mg protein The coupling process was also ascertained by addition of ADP. The venom suspension (VS) was prepared by homogenization of one venom sac in 2 ml of phosphate buffer. Fifty/A of VS was used in each experiment. In all cases the coupling was abolished. The activity of site III was reduced by 86%, of sites II and I by 100%. The reversibility of the venom damage was also estimated, after incubation with 25 tA, 50/al, 75/.d and 100 tal of VS and rinsing of the mitochondria. Treatment with 100 ~l, no recovery; with 25/al, 30% recovery. The impairment of oxidative phosphorylation is reflected in the electron microscope picture of the treated mitochondria, which appeared as swollen bags containing a few vesicles, instead of cristae. In conclusion, hornet venom damages the inner membrane of the mitochondrion. This can be explained by the presence of phosphorylase enzymes in the venom.
Effect o f tiger snake venom on the liver. E. A. BENNETTand I. F. COOMBE(Department of Medical Laboratory of Science, Queensland Institute of Technology, Brisbane, Australia). THE VENOMof the Australian mainland tiger snake (Notechis scutatus scutatas) contains three potent neurotoxic components. As well as acting on the neural synapse, they show powerful myolytic effects, one aspect of their participation in a general histolytic role of the venom in prey digestion. Vacuum-dried venom was dissolved in phosphate buffer (0.05 mole/l, pH 6.5). This was chromatographically fractionated by gel filtration on Sephadex G-150 and then by ion-exchange chromatography on carboxymethyl Sephadex C-25 to obtain five protein fractions (C-25a- e), the first, third and fourth being neurotoxic. Fractions were injected i.p. into mice and compared with whole venom. Tissues were removed for light and electron microscopy. All techniques were standard. Whole venom produced congestion of blood vessels, swelling of liver sinusoids and clearing of the
15
ABSTRACTS The abstracts are arranged alphabetically by first author. Only abstracts of work which was actually presented at the meeting are included in the following list.
Experimental studies on the enteropathogenicity ofVibrio fluvialis. R. K. AOARWALand S. C. SAr~YAL(Enteric Infection Unit, Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi221005, India). STRAINS of Vibriofluvialis isolated from diverse sources were tested for enterotoxigenicity in rabbit ileal loops, rabbit's skin and CHO cells and for enteroinvasiveness in rabbit's eye. Live cells and culture filtrates of all the strains of V. fluvialis tested were found to be enterotoxigenic. The enterotoxic factor was found to be heat labile, as evidenced by heat treatment, time course of fluid accumulation and negative suckling mouse assay. The outpouring of fluid into the loops was inhibited by chlorpromazine, indicating that the enterotoxin may act through mediation of cAMP. The tox gent(s) appears to be chromosomal, as the cells could not be cured of enterotoxicity by different methods used for elimination of plasmids. None of the strains tested could produce keratoconjunctivitis in rabbit's eye, indicating lack of invasive capability.
Clinico-pathological correlation in Russell's viper bite. MAUNOMAUNOAYE (Department of Surgery, Changi Hospital, Singapore). THIS presentation is based on personal clinical experience of 905 snake bite patients treated at Insein Hospital, Rangoon, Burma, over a ten year period. Among 905 patients, 434 victims were bitten by Russell's viper, which caused 145 deaths; post-mortem examination was performed on every fatal case. Russell's viper bite inflicts local reactions such as fang marks, oedema, blister, necrosis and even gangrene. More importantly, severe envenomation produces systemic manifestation such as shock, blood coagulation defect, spontaneous haemorrhages, acute renal failure, pituitary necrosis and even death. Cfinical features of Russell's viper bite will be illustrated and correlated with morbid and microscopic pathology. An outline of management of viper envenomation will also be presented.
Toxicology o f the plant Calotropis gigantea. J. AZARIAH, H. AT-~IAH, S. MALLIKESAN,M. V. SUMATHIand C. StmDARA~J (Department of Zoology and Centre for Advanced Study in Botany, University of Madras, Guindy Campus, Madras 600 025, India). IN TAMILNADUthe plant Calotropisgigantea is harvested and used for agricultural practices, especially in paddy cultivation. The plant is also used to cause abortion and homicide. Hence, a detailed literature survey and a few laboratory experiments were made. Information on the traditional medical use (non-scientific) and scientific work on the toxic nature of the plants was gathered. Paddy variety ADT 36, fresh water mussel Lamellidens marginalis and the mitotic index of root tips of Ailium cepa were used as biological and genetic screeing material to assess the toxicity of the plant extract. Aqueous extracts of the plant were made and other dilutions were made as required. It was found that the extract was toxic in high concentrations and with progressive dilutions seed germination was stimulated. Similarly, the extract was toxic to the survival of the mussels at high concentrations. The heart beat and the ciliary beat of the gill were seen to slow down at high concentrations. Lower dilutions accelerated both the functions. However, it was found that root germination of onion was inhibited due to cytotoxicity. There were both nuclear and chromosomal aberrations. Since people use the plants to cause abortion and in other treatments, it is recommended that the public be educated on its ill effects. A need to standardise the traditional use is suggested.
Biochemical assay using the aqueous extract o f Ptychodera flava (hemichordate) on the respiratory enzyme phenoloxiduse. P. S. R. BAI and J. AZARIAH(Department of Zoology, University of Madras, Guindy Campus, Madras 600 025, India). IT HAS BEEN established that the aqueous extract of Ptychodera flava can inhibit or stimulate the clevage of fertilized eggs of marine polychaetes. A study was made to assess the presence of physiologically active organic compounds. The effect of the extract was studied on the respiratory enzyme phenoloxidase of the surf crab Emerita emeritus. The extract was prepared on the recommendations outlined in a previous publication. On the
16
First A s i a - Pacific Congress
basis of the enzyme activity, the physiological activity of the extract was assayed at 480 nm using a spectrophotometer. The assay revealed that enzyme activity decreased soon after administration of the extract. The results indicated that there are pharmacologically and physiologically active compounds which can inhibit the enzyme activity depending upon its storage period, percentage composition and dosage adminstered. The Indian P. tiara has been characterized as containing iodocresol and ethylphenol compounds. There is no trace of indoles and hydroquinones. The phenol was found to be linked with bromine and iodine, whereas P. flava of western waters contained more bromophenols. It is concluded that the halometabolites may be responsible for the observed changes in enzyme activity.
Clinical features on 19,890 cases o f ciguatera (fish poisoning) in French Polynesia. R. A. BAGNIS (Institut Territorial de Recherces M6dicales Louis Malard6, BP 30, Papeete, Tahiti, Polyn6sie Fran~aise). THE EVALUATION of patients took place as part of an epidemiological program from 1964 to 1986. A standardized l-page questionnaire was used to obtain most of the clinical data. The information was gathered from patients in Society, Marquesas, Tuamotu, Gambier and the Austral Islands, in which ciguatera is known to be a relatively common problem. The majority of the questionnaire were completed by an examining physician or nurse. The most common symptoms could be grouped into four main categories: neurological, digestive, cardiovascular and generalized. From these, paraesthesia, arthralgia, myalgia, vertigo, ataxia, diarrhea, vomiting, nausea, bradycardia, hypotension, pruritus and asthenia account for the most frequent or most typical, with a frequency ranging from 90 to 15%. Approximately 77°70 of the patients became symptomatic less than 12 h after ingestion of the fish. The recovery period may vary from 72 h to several days, weeks or months. The rate of fatality was less than 1% 0, death generally occuring more from cardiac than from respiratory failure. Significant differences in certain symptoms and signs, as well as in evolutionary pattern, may be noted according to species of fish, susceptibility of patients and recurrence of illness. Being poisoned multiple times may result in a clinically more severe illness than disease observed in patients experiencing ciguatera for the first time. Hypersensitization to fish or seafood may be developed by some patients, resulting in allergic-like cutaneous, respiratory or digestive features.
Suppression o f oxidative phosphorylation in mitochondria by hornet venom. L. BARR-NEA,D. SHAHARand J. ISHAY (Suckler School of Medicine, Tel Aviv University, Ramat Aviv, Israel). THE PURPOSE of this work was to estimate the direct influence of hornet (Vespa orientalis) venom on liver mitochondria in vitro, since in vivo morphological studies had shown that the number of mitochondria and amount of their cristae was decreased in the liver of rats treated with hornet venom. The liver was homogenized and a mitochondrial preparation obtained after centrifugation at 10,000 g. The mitochondria were then incubated in different substrates for estimation of the activity of the different sites of electron transfer: malate or pyruvate for site I; succinate for site II; ascorbate for site III. Oxygen consumption was also estimated in an oxygen electrode. The results were expressed as nl oxygen atom/min/mg protein The coupling process was also ascertained by addition of ADP. The venom suspension (VS) was prepared by homogenization of one venom sac in 2 ml of phosphate buffer. Fifty/A of VS was used in each experiment. In all cases the coupling was abolished. The activity of site III was reduced by 86%, of sites II and I by 100%. The reversibility of the venom damage was also estimated, after incubation with 25 tA, 50/al, 75/.d and 100 tal of VS and rinsing of the mitochondria. Treatment with 100 ~l, no recovery; with 25/al, 30% recovery. The impairment of oxidative phosphorylation is reflected in the electron microscope picture of the treated mitochondria, which appeared as swollen bags containing a few vesicles, instead of cristae. In conclusion, hornet venom damages the inner membrane of the mitochondrion. This can be explained by the presence of phosphorylase enzymes in the venom.
Effect o f tiger snake venom on the liver. E. A. BENNETTand I. F. COOMBE(Department of Medical Laboratory of Science, Queensland Institute of Technology, Brisbane, Australia). THE VENOMof the Australian mainland tiger snake (Notechis scutatus scutatas) contains three potent neurotoxic components. As well as acting on the neural synapse, they show powerful myolytic effects, one aspect of their participation in a general histolytic role of the venom in prey digestion. Vacuum-dried venom was dissolved in phosphate buffer (0.05 mole/l, pH 6.5). This was chromatographically fractionated by gel filtration on Sephadex G-150 and then by ion-exchange chromatography on carboxymethyl Sephadex C-25 to obtain five protein fractions (C-25a- e), the first, third and fourth being neurotoxic. Fractions were injected i.p. into mice and compared with whole venom. Tissues were removed for light and electron microscopy. All techniques were standard. Whole venom produced congestion of blood vessels, swelling of liver sinusoids and clearing of the