DOCSLIB.ORG

Cloning and Characterization of Novel Snake Venom Proteins That Block Smooth Muscle Contraction

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cloning and Characterization of Novel Snake Venom Proteins That Block Smooth Muscle Contraction Eur. J. Biochem. 269, 2708–2715 (2002) Ó FEBS 2002 doi:10.1046/j.1432-1033.2002.02940.x Cloning and characterization of novel snake venom proteins that block smooth muscle contraction Yasuo Yamazaki1, Hisashi Koike1, Yusuke Sugiyama1, Kazuko Motoyoshi1, Taeko Wada1, Shigeru Hishinuma2, Mitsuo Mita2 and Takashi Morita1 Departments of 1Biochemistry; and 2Pharmacodynamics, Meiji Pharmaceutical University, Tokyo, Japan In this study, we isolated a 25-kDa novel snake venom caffeine-stimulated contraction. Furthermore, we isolated protein, designated ablomin, from the venom of the Japan- three other proteins from snake venoms that are homolog- ese Mamushi snake (Agkistrodon blomhoffi). The amino-acid ous to ablomin and cloned the corresponding cDNAs. Two sequence of this protein was determined by peptide of these homologous proteins, triflin and latisemin, also sequencing and cDNA cloning. The deduced sequence inhibited high K+-induced contraction of the artery. These showed high similarity to helothermine from the Mexican results indicate that several snake venoms contain novel beaded lizard (Heloderma horridum horridum), which blocks proteins with neurotoxin-like activity. voltage-gated calcium and potassium channels, and ryano- Keywords: snake venom; neurotoxin; helothermine; cysteine- dine receptors. Ablomin blocked contraction of rat tail rich secretory proteins; ablomin. arterial smooth muscle elicited by high K+-induced depolarization in the 0.1–1 lM range, but did not block Over the past 30 years, a plethora of toxins have been homologous to Kunitz-type serine protease inhibitors and isolated from poisonous organisms, such as snakes, scorpi- act primarily by blocking neuronal K+ channels [30,31]. In ons, spiders, and micro-organisms. These natural toxins use contrast to the neurotoxin-rich venom from Elapidae a variety of approaches to arrest the homeostatic mecha- snakes, the venom from other deadly snakes, including nisms of other living organisms, including disruption of Viperidae and Colubridae snakes, contain surprisingly few intracellular signal transduction and cytoskeleton organiza- neurotoxins, although some neurotoxic phospholipases tion [1–4], and activation or inhibition of blood coagulation have been discovered [32–36]. factors [5–10]. Toxins that block synaptic transmission, In this report, we describe the isolation of a novel protein, called neurotoxins, are widely distributed in venoms. These ablomin, from the venom of the Japanese Mamushi snake toxins include the conotoxins from cone snails, agatoxins (Agkistrodon blomhoffi, a member of the Viperidae family). from spiders, and scorpion toxins [11–16]. These toxins exert When applied to arterial smooth muscle preparations from their potentially lethal effects by specifically and potently rat-tails, ablomin blocks K+-stimulated contraction. This blocking a variety of ion channels, including those that effect is similar to that resulting from application of conduct Na+,K+,andCa2+. Therefore, neurotoxins have calciseptine, a well-characterized neurotoxin from black been employed as useful tools to investigate the structure mamba (Dendroaspis polylepis polylepis).Calciseptineisa and function of these ion channels [17–20]. A large number known blocker of L-type Ca2+ channels, a property that of neurotoxin families have also been found in the venom of underlies its ability to block K+-induced contractions of Elapidae snakes. These toxins, the a-neurotoxins [21] aortic smooth muscle and spontaneous contractions of (represented by a-bungarotoxin [22,23], a-cobratoxin uterine smooth muscle [37]. Furthermore, we demonstrate [24–27], and erabutoxin [28,29])potently and specifically that several snake venoms contain ablomin-like proteins, prevent nicotinic acetylcholine receptor activation. A second which may constitute a novel venom protein family. family of snake venom neurotoxins, the dendrotoxins, are EXPERIMENTAL PROCEDURES Correspondence to T. Morita, Department of Biochemistry, Materials Meiji Pharmaceutical University, 2-522-1, Noshio, Kiyose, Tokyo 204-8588, Japan, The lyophilized venom of A. blomhoffi was a kind gift from Fax/Tel.: + 81 424 95 8479, S. Iwanaga (The Chemo-Sero-Therapeutic Research E-mail: [email protected] Institute, Kumamoto, Japan)[38]. Other snake venoms Abbreviations: CRISP, cysteine-rich secretory protein; HLTX, and venom glands were purchased from the Japan Snake helothermine; PsTx, pseudechetoxin; CAP, CRISPs Antigen 5 Institute (Gunma, Japan). Superdex 75 pg and 200 pg, proteins, and Pathogenesis-related proteins. SP–Sepharose High Performance, and Q-Sepharose Fast Note: the nucleotide sequences reported here have been submitted to Flow columns were from Amersham–Pharmacia Biotech. GenBank database (tigrin, AY093955; ablomin, AF384218; triflin, The Vydac Protein & Peptide C18 HPLC column and the AF384219; latisemin, AF384220). COSMOSIL 5C18 AR-300 HPLC column were the (Received 21 December 2001, revised 12 April 2002, products of JASCO (Tokyo, Japan)and Nacalai Tesque accepted 18 April 2002) (Kyoto, Japan), respectively. Endoprotease Lys-C was Ó FEBS 2002 Novel proteins in snake venoms (Eur. J. Biochem. 269)2709 purchased from Seikagaku Corporation (Tokyo, Japan). Amino-acid sequence analysis Other chemicals were of analytical grade (Sigma–Aldrich, Amersham–Pharmacia Biotech., Wako Pure Chemical Ind. Proteins were reduced for 3 h at room temperature with and Kanto Chemical Co.). 20 mM dithiothreitol in the presence of 0.5 M Tris/HCl, pH 8.5, 6 M guanidine hydrochloride, and 2 mM EDTA in a volume of 0.5 mL. Three microliters of 4-vinylpyridine Purification of proteins were then added, and alkylation was allowed to proceed Tigrin was isolated from the extract of Duvernoy’s glands of for 3 h at room temperature. The S-pyridylethylated Rhabdophis tigrinus tigrinus. Ten Duvernoy’s glands were proteins were separated from the reagents by C18 broken into small pieces after freezing in liquid nitrogen, reverse-phase HPLC, and the amino-acid sequence was and then extracted in 30 mL of 50 mM Tris/HCl pH 8.0 for determined by sequencing the peptides obtained by diges- 4hat4°C. After ultracentrifugation, the supernatant was tion with endoprotease Lys-C. All the samples were applied onto Q-Sepharose Fast Flow column (1.6 · 10 cm) analyzed on Applied Biosystems protein sequencers (mod- in the same buffer, and eluted with a linear gradient from 0 els 473 A and 477). to 0.5 M NaCl. A major peak eluted at 0.05 M NaCl, which was subsequently purified by chromatography on Superdex cDNA cloning of proteins 200 pg (2.6 · 60 cm column). Ablomin was purified by three successive chromato- The cDNAs encoding tigrin, ablomin, and latisemin were graphic steps. Five hundred milligrams of lyophilized obtained using the RT-PCR method. Typically, venom A. blomhoffi venom was dissolved in 3 mL of 20 mM gland total RNA was isolated from the venom gland with TM imidazole/HCl pH 6.8 containing 0.2 M NaCl, and insol- ISOGEN (Wako Pure Chemical Industries, Japan) uble materials were removed by centrifugation and according to the manufacturer’s protocol. 5¢ and 3¢ RACE filtration (0.22 lm). The filtrate was loaded onto a were carried out to determine the nucleotide sequence of the Superdex 75 pg column (2.6 · 60 cm)and eluted with 5¢ and 3¢ end cDNAs with the SMARTTM RACE cDNA the same buffer. The ablomin fractions from two gel amplification kit (Clontech). The amino-acid sequences of filtration runs (a total of 1000 mg of snake venom)were peptides derived from purified proteins were used to design pooled and dialyzed against 50 mM Tris/HCl, pH 8.0, and degenerate primers. For the first amplification of tigrin and applied to the Q-Sepahrose Fast Flow column latisemin cDNA, degenerate primers were used for both (1.6 · 15 cm). The column was eluted with a linear sense and antisense primers. For ablomin cDNA, PCR was gradient of NaCl from 0 to 0.4 M at a flow rate of performed with single degenerate primer (sense or antisense) 2mLÆmin)1. Chromatographic fractions containing and an primer recognizing an adaptor sequence that had ablomin were then dialyzed against 20 mM imidazole- been attached to the 5¢ or 3¢ endofcDNAs.Inthecaseof HCl, pH 6.0, and fractionated on a SP–Sepharose High triflin, PCR was carried out using habu cDNA library as a Performance column (1.6 · 11 cm). This column was template [40] with a degenerate primer and an adaptor developed with a linear gradient of NaCl in the imidazole primer. The PCR products were subcloned into the pGEM )1 buffer (0–0.4 M,2mLÆmin ). T-easy vector (tigrin and triflin)or pUC19 vector (ablomin For the purification of triflin, 500 mg of the venom of and latisemin)and sequenced with the DSQ 2000 L DNA Trimeresurus flavoviridis was applied to the SP–Sepharose sequencer (Shimadzu, Japan). Primers used this study are Fast Flow column (1.6 · 30 cm)with 10 m M phosphate described as follows: tigrin, sense 5¢-AA(C,T)GT(A,C,G,T) buffer, pH 6.8, and eluted with a linear gradient from 0 to GA(C,T)TT(C,T)AA(C,T)TC(A,C,G,T)GA(A,G)TC-3¢ 0.15 M NaCl, as described previously [39]. Fractions con- (corresponding to amino acids 1–8 in tigrin)and antisense taining triflin were detected by Western blotting using anti- 5¢-(A,G)TT(A,G)CA(A,G)TT(A,G)TT(A,G)TA(A,G)TC tigrin serum. These fractions were pooled and fractionated (A,G)TC-3¢ (corresponding to amino acids 187–193 in on Superdex 75 pg (2.6 · 60 cm)in a 50-m M Tris/HCl, tigrin); ablomin, sense 5¢-GGCCATTA(C,T)ACTCAG(A,G) pH 8.0, containing 0.2 M NaCl. Finally, triflin was purified T(A,G)G-3¢ (corresponding to amino acids 114–120 in by chromatography on a Blue-Sepharose Fast Flow column ablomin)and antisense 5 ¢-C(C,T)A(C,T)CTGAGT(A,G) (1.6 · 15.5 cm)in 50 m M Tris/HCl, pH 8.0, which was TAATGGCC-3¢ (corresponding to amino acids 114–120 eluted with a linear gradient from 0 to 0.5 M NaCl.
Load more

Recommended publications
  • Snake Charming and the Exploitation of Snakes in Morocco
    Snake charming and the exploitation of snakes in Morocco J UAN M. PLEGUEZUELOS,MÓNICA F ERICHE,JOSÉ C. BRITO and S OUMÍA F AHD Abstract Traditional activities that potentially threaten bio- also for clothing, tools, medicine and pets, as well as in diversity represent a challenge to conservationists as they try magic and religious activities (review in Alves & Rosa, to reconcile the cultural dimensions of such activities. ). Vertebrates, particularly reptiles, have frequently Quantifying the impact of traditional activities on biodiver- been used for traditional medicine. Alves et al. () iden- sity is always helpful for decision making in conservation. In tified reptile species ( families, genera) currently the case of snake charming in Morocco, the practice was in- used in traditional folk medicine, % of which are included troduced there years ago by the religious order the on the IUCN Red List (IUCN, ) and/or the CITES Aissawas, and is now an attraction in the country’s growing Appendices (CITES, ). Among the reptile species tourism industry. As a consequence wild snake populations being used for medicine, % are snakes. may be threatened by overexploitation. The focal species for Snakes have always both fascinated and repelled people, snake charming, the Egyptian cobra Naja haje, is undergo- and the reported use of snakes in magic and religious activ- ing both range and population declines. We estimated the ities is global (Alves et al., ). The sacred role of snakes level of exploitation of snakes based on field surveys and may be related to a traditional association with health and questionnaires administered to Aissawas during – eternity in some cultures (Angeletti et al., ) and many , and compared our results with those of a study con- species are under pressure from exploitation as a result ducted years previously.
    [Show full text]
  • Comprehensive Snake Venomics of the Okinawa Habu Pit Viper, Protobothrops Flavoviridis, by Complementary Mass Spectrometry-Guided Approaches
    Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 6 July 2018 doi:10.20944/preprints201807.0123.v1 Peer-reviewed version available at Molecules 2018, 23, 1893; doi:10.3390/molecules23081893 Article Comprehensive Snake Venomics of the Okinawa Habu Pit Viper, Protobothrops Flavoviridis, by Complementary Mass Spectrometry-Guided Approaches Maik Damm 1,┼, Benjamin-Florian Hempel 1,┼, Ayse Nalbantsoy 2 and Roderich D. Süssmuth 1,* 1 Department of Chemistry, Technische Universität Berlin, 10623 Berlin, Germany; [email protected] (M.D.); [email protected] (B.-F.H.) 2 Department of Bioengineering, Ege University, 35100 Izmir, Turkey; [email protected] (A.N.) * Correspondence: [email protected]; Tel.: +49-30-314-24205 ┼ These authors contributed equal to this work Abstract: The Asian world is home to a multitude of venomous and dangerous snakes, which are attributed to various medical effects used in the preparation of traditional snake tinctures and alcoholics, like the Japanese snake wine, named Habushu. The aim of this work was to perform the first quantitative proteomic analysis of the Protobothrops flavoviridis pit viper venom. Accordingly, the venom was analyzed by complimentary bottom-up and top-down mass spectrometry techniques. The mass spectrometry-based snake venomics approach revealed that more than half of the venom is composed of different phospholipases A2 (PLA2). The combination with an intact mass profiling led to the identification of the three main Habu PLA2s. Furthermore, nearly one-third of the total venom consists of snake venom metalloproteinases and disintegrins, and several minor represented toxins families were detected: CTL, CRISP, svSP, LAAO, PDE and 5’-nucleotidase.
    [Show full text]
  • WHO Guidance on Management of Snakebites
    GUIDELINES FOR THE MANAGEMENT OF SNAKEBITES 2nd Edition GUIDELINES FOR THE MANAGEMENT OF SNAKEBITES 2nd Edition 1. 2. 3. 4. ISBN 978-92-9022- © World Health Organization 2016 2nd Edition All rights reserved. Requests for publications, or for permission to reproduce or translate WHO publications, whether for sale or for noncommercial distribution, can be obtained from Publishing and Sales, World Health Organization, Regional Office for South-East Asia, Indraprastha Estate, Mahatma Gandhi Marg, New Delhi-110 002, India (fax: +91-11-23370197; e-mail: publications@ searo.who.int). The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement. The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsed or recommended by the World Health Organization in preference to others of a similar nature that are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguished by initial capital letters. All reasonable precautions have been taken by the World Health Organization to verify the information contained in this publication. However, the published material is being distributed without warranty of any kind, either expressed or implied. The responsibility for the interpretation and use of the material lies with the reader. In no event shall the World Health Organization be liable for damages arising from its use.
    [Show full text]
  • Mating Season of the Japanese Mamushi, Agkistrodon Blomhoffii Blomhoffii (Viperidae: Crotalinae), in Southern Kyushu, Japan: Relation with Female Ovarian Development
    Japanese Journal of Herpetology 16(2): 42-48., Dec. 1995 (C)1995 by The HerpetologicalSociety of Japan Mating Season of the Japanese Mamushi, Agkistrodon blomhoffii blomhoffii (Viperidae: Crotalinae), in Southern Kyushu, Japan: Relation with Female Ovarian Development KIYOSHI ISOGAWA AND MASAHIDE KATO Abstract: Mating in the southern Kyushu population of the Japanese mamushi, Agkistrodon blomhoffii blomhoffii, occurred in August and September. The largest ovarian follicles of mated females were 6mm or more in length, whereas those of unmated females were usually less than 6mm in length. In addition, body mass was significantly greater in mated females than in unmated ones. These suggest that in females of the Japanese mamushi, mating is strongly associated with ovarian follicle development and body mass. Key words: Mamushi; Agkistrodon blomhoffii blomhoffii; Mating season; Follicle development; Sexual receptivity Many reports have been published on the the southern Kyushu, Japan and to examine the mating season of snakes of the genus relation of the occurrence of mating with Agkistrodon (Allen and Swindell, 1948; Behler ovarian development, body size, and nutritional and King, 1979; Burchfield, 1982; Fitch, 1960; conditions in females. Gloyd, 1934; Huang et al., 1990; Laszlo, 1979; Li et al., 1992; Wright and Wright, 1957). MATERIALSAND METHODS Fukada (1972) surmised that mating in the Animals. -Forty-five mature females and the Japanese mamushi, Agkistrodon blomhoffii same number of mature males of Agkistrodon blomhoffii, as well as in many of the colubrids of blomhoffii blomhoffii were used. All were cap- Japan, occurs in May and June. No data, tured in Kagoshima Prefecture, Japan during however, have been obtained for the mating the summer of 1989.
    [Show full text]
  • An in Vivo Examination of the Differences Between Rapid
    www.nature.com/scientificreports OPEN An in vivo examination of the diferences between rapid cardiovascular collapse and prolonged hypotension induced by snake venom Rahini Kakumanu1, Barbara K. Kemp-Harper1, Anjana Silva 1,2, Sanjaya Kuruppu3, Geofrey K. Isbister 1,4 & Wayne C. Hodgson1* We investigated the cardiovascular efects of venoms from seven medically important species of snakes: Australian Eastern Brown snake (Pseudonaja textilis), Sri Lankan Russell’s viper (Daboia russelii), Javanese Russell’s viper (D. siamensis), Gaboon viper (Bitis gabonica), Uracoan rattlesnake (Crotalus vegrandis), Carpet viper (Echis ocellatus) and Puf adder (Bitis arietans), and identifed two distinct patterns of efects: i.e. rapid cardiovascular collapse and prolonged hypotension. P. textilis (5 µg/kg, i.v.) and E. ocellatus (50 µg/kg, i.v.) venoms induced rapid (i.e. within 2 min) cardiovascular collapse in anaesthetised rats. P. textilis (20 mg/kg, i.m.) caused collapse within 10 min. D. russelii (100 µg/kg, i.v.) and D. siamensis (100 µg/kg, i.v.) venoms caused ‘prolonged hypotension’, characterised by a persistent decrease in blood pressure with recovery. D. russelii venom (50 mg/kg and 100 mg/kg, i.m.) also caused prolonged hypotension. A priming dose of P. textilis venom (2 µg/kg, i.v.) prevented collapse by E. ocellatus venom (50 µg/kg, i.v.), but had no signifcant efect on subsequent addition of D. russelii venom (1 mg/kg, i.v). Two priming doses (1 µg/kg, i.v.) of E. ocellatus venom prevented collapse by E. ocellatus venom (50 µg/kg, i.v.). B. gabonica, C. vegrandis and B.
    [Show full text]
  • (Gloydius Blomhoffii) Antivenom in Japan, Korea, and China
    Jpn. J. Infect. Dis., 59, 20-24, 2006 Original Article Standardization of Regional Reference for Mamushi (Gloydius blomhoffii) Antivenom in Japan, Korea, and China Tadashi Fukuda*, Masaaki Iwaki, Seung Hwa Hong1, Ho Jung Oh1, Zhu Wei2, Kazunori Morokuma3, Kunio Ohkuma3, Lei Dianliang4, Yoshichika Arakawa and Motohide Takahashi Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, Tokyo 208-0011; 3First, Production Department, Chemo-Sero-Therapeutic Research Institute, Kumamoto 860-8568, Japan; 1Korea Food and Drug Administration, Soul 122-704, Korea; 2Shanghai Institute of Biological Products, Shanghai 200052; and 4Department of Serum, National Institute for the Control of Pharmaceutical and Biological Products, Beijing 10050, People’s Republic of China (Received June 27, 2005. Accepted November 11, 2005) SUMMARY: The mamushi (Gloydius blomhoffii) snakes that inhabit Japan, Korea, and China produce venoms with similar serological characters to each other. Individual domestic standard mamushi antivenoms have been used for national quality control (potency testing) of mamushi antivenom products in these countries, because of the lack of an international standard material authorized by the World Health Organization. This precludes comparison of the results of product potency testing among countries. We established a regional reference antivenom for these three Asian countries. This collaborative study indicated that the regional reference mamushi antivenom has an anti-lethal titer of 33,000 U/vial and anti-hemorrhagic titer of 36,000 U/vial. This reference can be used routinely for quality control, including national control of mamushi antivenom products. reference antivenom. INTRODUCTION In the present study, the potency of a candidate regional Snakebites are a threat to human life in areas inhabited by reference mamushi antivenom produced by Shanghai Insti- poisonous snakes.
    [Show full text]
  • Toxicology in Antiquity
    TOXICOLOGY IN ANTIQUITY Other published books in the History of Toxicology and Environmental Health series Wexler, History of Toxicology and Environmental Health: Toxicology in Antiquity, Volume I, May 2014, 978-0-12-800045-8 Wexler, History of Toxicology and Environmental Health: Toxicology in Antiquity, Volume II, September 2014, 978-0-12-801506-3 Wexler, Toxicology in the Middle Ages and Renaissance, March 2017, 978-0-12-809554-6 Bobst, History of Risk Assessment in Toxicology, October 2017, 978-0-12-809532-4 Balls, et al., The History of Alternative Test Methods in Toxicology, October 2018, 978-0-12-813697-3 TOXICOLOGY IN ANTIQUITY SECOND EDITION Edited by PHILIP WEXLER Retired, National Library of Medicine’s (NLM) Toxicology and Environmental Health Information Program, Bethesda, MD, USA Academic Press is an imprint of Elsevier 125 London Wall, London EC2Y 5AS, United Kingdom 525 B Street, Suite 1650, San Diego, CA 92101, United States 50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom Copyright r 2019 Elsevier Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/permissions. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein).
    [Show full text]
  • Venomics of Trimeresurus (Popeia) Nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights Into Venom Proteome, Toxicity and Neutralization of Antivenom
    toxins Article Venomics of Trimeresurus (Popeia) nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights into Venom Proteome, Toxicity and Neutralization of Antivenom Choo Hock Tan 1,*, Kae Yi Tan 2 , Tzu Shan Ng 2, Evan S.H. Quah 3 , Ahmad Khaldun Ismail 4 , Sumana Khomvilai 5, Visith Sitprija 5 and Nget Hong Tan 2 1 Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; 2 Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; [email protected] (K.Y.T.); [email protected] (T.S.N.); [email protected] (N.H.T.) 3 School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia; [email protected] 4 Department of Emergency Medicine, Universiti Kebangsaan Malaysia Medical Centre, 56000 Kuala Lumpur, Malaysia; [email protected] 5 Thai Red Cross Society, Queen Saovabha Memorial Institute, Bangkok 10330, Thailand; [email protected] (S.K.); [email protected] (V.S.) * Correspondence: [email protected] Received: 31 December 2018; Accepted: 30 January 2019; Published: 6 February 2019 Abstract: Trimeresurus nebularis is a montane pit viper that causes bites and envenomation to various communities in the central highland region of Malaysia, in particular Cameron’s Highlands. To unravel the venom composition of this species, the venom proteins were digested by trypsin and subjected to nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) for proteomic profiling. Snake venom metalloproteinases (SVMP) dominated the venom proteome by 48.42% of total venom proteins, with a characteristic distribution of P-III: P-II classes in a ratio of 2:1, while P-I class was undetected.
    [Show full text]
  • Ecology, Behavior and Conservation of the Japanese Mamushi Snake, Gloydius Blomhoffii: Variation in Compromised and Uncompromised Populations
    ECOLOGY, BEHAVIOR AND CONSERVATION OF THE JAPANESE MAMUSHI SNAKE, GLOYDIUS BLOMHOFFII: VARIATION IN COMPROMISED AND UNCOMPROMISED POPULATIONS By KIYOSHI SASAKI Bachelor of Arts/Science in Zoology Oklahoma State University Stillwater, OK 1999 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY December, 2006 ECOLOGY, BEHAVIOR AND CONSERVATION OF THE JAPANESE MAMUSHI SNAKE, GLOYDIUS BLOMHOFFII: VARIATION IN COMPROMISED AND UNCOMPROMISED POPULATIONS Dissertation Approved: Stanley F. Fox Dissertation Adviser Anthony A. Echelle Michael W. Palmer Ronald A. Van Den Bussche A. Gordon Emslie Dean of the Graduate College ii ACKNOWLEDGMENTS I sincerely thank the following people for their significant contribution in my pursuit of a Ph.D. degree. I could never have completed this work without their help. Dr. David Duvall, my former mentor, helped in various ways until the very end of his career at Oklahoma State University. This study was originally developed as an undergraduate research project under Dr. Duvall. Subsequently, he accepted me as his graduate student and helped me expand the project to this Ph.D. research. He gave me much key advice and conceptual ideas for this study. His encouragement helped me to get through several difficult times in my pursuit of a Ph.D. degree. He also gave me several books as a gift and as an encouragement to complete the degree. Dr. Stanley Fox kindly accepted to serve as my major adviser after Dr. Duvall’s departure from Oklahoma State University and involved himself and contributed substantially to this work, including analysis and editing.
    [Show full text]
  • Venom Proteomics and Antivenom Neutralization for the Chinese
    www.nature.com/scientificreports OPEN Venom proteomics and antivenom neutralization for the Chinese eastern Russell’s viper, Daboia Received: 27 September 2017 Accepted: 6 April 2018 siamensis from Guangxi and Taiwan Published: xx xx xxxx Kae Yi Tan1, Nget Hong Tan1 & Choo Hock Tan2 The eastern Russell’s viper (Daboia siamensis) causes primarily hemotoxic envenomation. Applying shotgun proteomic approach, the present study unveiled the protein complexity and geographical variation of eastern D. siamensis venoms originated from Guangxi and Taiwan. The snake venoms from the two geographical locales shared comparable expression of major proteins notwithstanding variability in their toxin proteoforms. More than 90% of total venom proteins belong to the toxin families of Kunitz-type serine protease inhibitor, phospholipase A2, C-type lectin/lectin-like protein, serine protease and metalloproteinase. Daboia siamensis Monovalent Antivenom produced in Taiwan (DsMAV-Taiwan) was immunoreactive toward the Guangxi D. siamensis venom, and efectively neutralized the venom lethality at a potency of 1.41 mg venom per ml antivenom. This was corroborated by the antivenom efective neutralization against the venom procoagulant (ED = 0.044 ± 0.002 µl, 2.03 ± 0.12 mg/ml) and hemorrhagic (ED50 = 0.871 ± 0.159 µl, 7.85 ± 3.70 mg/ ml) efects. The hetero-specifc Chinese pit viper antivenoms i.e. Deinagkistrodon acutus Monovalent Antivenom and Gloydius brevicaudus Monovalent Antivenom showed negligible immunoreactivity and poor neutralization against the Guangxi D. siamensis venom. The fndings suggest the need for improving treatment of D. siamensis envenomation in the region through the production and the use of appropriate antivenom. Daboia is a genus of the Viperinae subfamily (family: Viperidae), comprising a group of vipers commonly known as Russell’s viper native to the Old World1.
    [Show full text]
  • Injuries and Envenomation by Exotic Pets in Hong Kong Vember CH Ng, Albert CH Lit, of Wong *, ML Tse, HT Fung
    Original Article Injuries and envenomation by exotic pets in Hong Kong Vember CH Ng, Albert CH Lit, OF Wong *, ML Tse, HT Fung ABSTRACT effects, and six cases with mild effects. All major effects were related to venomous snakebites. There Exotic pets are increasingly popular Introduction: were no mortalities. in Hong Kong and include fish, amphibians, reptiles, and arthropods. Some of these exotic Conclusion: All human injuries from exotic pets animals are venomous and may cause injuries to arose from reptiles, scorpions, and fish. All cases of and envenomation of their owners. The clinical major envenomation were inflicted by snakes. experience of emergency physicians in the management of injuries and envenomation by these exotic animals is limited. We reviewed the clinical Hong Kong Med J 2018;24:48–55 features and outcomes of injuries and envenomation DOI: 10.12809/hkmj176984 by exotic pets recorded by the Hong Kong Poison 1 VCH Ng, FHKCEM, FHKAM (Emergency Medicine) Information Centre. 2 ACH Lit, FRCSEd, FHKAM (Emergency Medicine) Methods: We retrospectively retrieved and reviewed 2 OF Wong *, FHKAM (Anaesthesiology), FHKAM (Emergency Medicine) 1 cases of injuries and envenomation by exotic pets ML Tse, FHKCEM, FHKAM (Emergency Medicine) 3 HT Fung, recorded by the Hong Kong Poison Information FRCSEd, FHKAM (Emergency Medicine) Centre from 1 July 2008 to 31 March 2017. 1 Hong Kong Poison Information Centre, United Christian Hospital, Kwun Results: There were 15 reported cases of injuries Tong, Hong Kong 2 Accident and Emergency Department, North Lantau Hospital, Tung and envenomation by exotic pets during the study Chung, Lantau, Hong Kong period, including snakebite (n=6), fish sting (n=4), 3 Accident and Emergency Department, Tuen Mun Hospital, Tuen Mun, This article was scorpion sting (n=2), lizard bite (n=2), and turtle Hong Kong published on 5 Jan bite (n=1).
    [Show full text]
  • Neurotoxicity in Snakebite—The Limits of Our Knowledge
    Review Neurotoxicity in Snakebite—The Limits of Our Knowledge Udaya K. Ranawaka1*, David G. Lalloo2, H. Janaka de Silva1 1 Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka, 2 Liverpool School of Tropical Medicine, Liverpool, United Kingdom been well described with pit vipers (family Viperidae, subfamily Abstract: Snakebite is classified by the WHO as a Crotalinae) such as rattlesnakes (Crotalus spp.) [58–67]. Although neglected tropical disease. Envenoming is a significant considered relatively less common with true vipers (family public health problem in tropical and subtropical regions. Viperidae, subfamily Viperinae), neurotoxicity is well recognized Neurotoxicity is a key feature of some envenomings, and in envenoming with Russell’s viper (Daboia russelii) in Sri Lanka and there are many unanswered questions regarding this South India [9,68–75], the asp viper (Vipera aspis) [76–82], the manifestation. Acute neuromuscular weakness with respi- adder (Vipera berus) [83–85], and the nose-horned viper (Vipera ratory involvement is the most clinically important ammodytes) [86,87]. neurotoxic effect. Data is limited on the many other acute neurotoxic manifestations, and especially delayed Acute neuromuscular paralysis is the main type of neurotoxicity neurotoxicity. Symptom evolution and recovery, patterns and is an important cause of morbidity and mortality related to of weakness, respiratory involvement, and response to snakebite. Mechanical ventilation, intensive care, antivenom antivenom and acetyl cholinesterase inhibitors are vari- treatment, other ancillary care, and prolonged hospital stays all able, and seem to depend on the snake species, type of contribute to a significant cost of provision of care. And ironically, neurotoxicity, and geographical variations. Recent data snakebite is common in resource-poor countries that can ill afford have challenged the traditional concepts of neurotoxicity such treatment costs.
    [Show full text]