Venom Week 2012 4Th International Scientific Symposium on All Things Venomous
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Phylogenetic Diversity, Habitat Loss and Conservation in South
Diversity and Distributions, (Diversity Distrib.) (2014) 20, 1108–1119 BIODIVERSITY Phylogenetic diversity, habitat loss and RESEARCH conservation in South American pitvipers (Crotalinae: Bothrops and Bothrocophias) Jessica Fenker1, Leonardo G. Tedeschi1, Robert Alexander Pyron2 and Cristiano de C. Nogueira1*,† 1Departamento de Zoologia, Universidade de ABSTRACT Brasılia, 70910-9004 Brasılia, Distrito Aim To analyze impacts of habitat loss on evolutionary diversity and to test Federal, Brazil, 2Department of Biological widely used biodiversity metrics as surrogates for phylogenetic diversity, we Sciences, The George Washington University, 2023 G. St. NW, Washington, DC 20052, study spatial and taxonomic patterns of phylogenetic diversity in a wide-rang- USA ing endemic Neotropical snake lineage. Location South America and the Antilles. Methods We updated distribution maps for 41 taxa, using species distribution A Journal of Conservation Biogeography models and a revised presence-records database. We estimated evolutionary dis- tinctiveness (ED) for each taxon using recent molecular and morphological phylogenies and weighted these values with two measures of extinction risk: percentages of habitat loss and IUCN threat status. We mapped phylogenetic diversity and richness levels and compared phylogenetic distances in pitviper subsets selected via endemism, richness, threat, habitat loss, biome type and the presence in biodiversity hotspots to values obtained in randomized assemblages. Results Evolutionary distinctiveness differed according to the phylogeny used, and conservation assessment ranks varied according to the chosen proxy of extinction risk. Two of the three main areas of high phylogenetic diversity were coincident with areas of high species richness. A third area was identified only by one phylogeny and was not a richness hotspot. Faunal assemblages identified by level of endemism, habitat loss, biome type or the presence in biodiversity hotspots captured phylogenetic diversity levels no better than random assem- blages. -
Distinct Muscarinic Acetylcholine Receptor Subtypes Contribute to Stability and Growth, but Not Compensatory Plasticity, of Neuromuscular Synapses
14942 • The Journal of Neuroscience, November 25, 2009 • 29(47):14942–14955 Development/Plasticity/Repair Distinct Muscarinic Acetylcholine Receptor Subtypes Contribute to Stability and Growth, But Not Compensatory Plasticity, of Neuromuscular Synapses Megan C. Wright,1 Srilatha Potluri,1 Xueyong Wang,2 Eva Dentcheva,1 Dinesh Gautam,3 Alan Tessler,1,4 Ju¨rgen Wess,3 Mark M. Rich,2 and Young-Jin Son1 1Department of Neurobiology and Anatomy, Drexel University College of Medicine, Philadelphia, Pennsylvania 19129, 2Department of Neuroscience, Cell Biology, and Physiology, Wright State University, Dayton, Ohio 45435, 3Molecular Signaling Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892, and 4Department of Neurology, Department of Veterans Affairs Hospital, Philadelphia, Pennsylvania 19104 Muscarinic acetylcholine receptors (mAChRs) modulate synaptic function, but whether they influence synaptic structure remains un- known. At neuromuscular junctions (NMJs), mAChRs have been implicated in compensatory sprouting of axon terminals in paralyzed or denervated muscles. Here we used pharmacological and genetic inhibition and localization studies of mAChR subtypes at mouse NMJs to demonstrate their roles in synaptic stability and growth but not in compensatory sprouting. M2 mAChRs were present solely in motor neurons,whereasM1 ,M3 ,andM5 mAChRswereassociatedwithSchwanncellsand/ormusclefibers.BlockadeofallfivemAChRsubtypes with atropine evoked pronounced effects, -
Muscarinic Acetylcholine Type 1 Receptor Activity Constrains Neurite Outgrowth by Inhibiting Microtubule Polymerization and Mito
fnins-12-00402 June 26, 2018 Time: 12:46 # 1 ORIGINAL RESEARCH published: 26 June 2018 doi: 10.3389/fnins.2018.00402 Muscarinic Acetylcholine Type 1 Receptor Activity Constrains Neurite Outgrowth by Inhibiting Microtubule Polymerization and Mitochondrial Trafficking in Adult Sensory Neurons Mohammad G. Sabbir1*, Nigel A. Calcutt2 and Paul Fernyhough1,3 1 Division of Neurodegenerative Disorders, St. Boniface Hospital Research Centre, Winnipeg, MB, Canada, 2 Department of Pathology, University of California, San Diego, San Diego, CA, United States, 3 Department of Pharmacology and Therapeutics, University of Manitoba, Winnipeg, MB, Canada The muscarinic acetylcholine type 1 receptor (M1R) is a metabotropic G protein-coupled Edited by: receptor. Knockout of M1R or exposure to selective or specific receptor antagonists Roberto Di Maio, elevates neurite outgrowth in adult sensory neurons and is therapeutic in diverse University of Pittsburgh, United States models of peripheral neuropathy. We tested the hypothesis that endogenous M1R Reviewed by: activation constrained neurite outgrowth via a negative impact on the cytoskeleton Roland Brandt, University of Osnabrück, Germany and subsequent mitochondrial trafficking. We overexpressed M1R in primary cultures Rick Dobrowsky, of adult rat sensory neurons and cell lines and studied the physiological and The University of Kansas, United States molecular consequences related to regulation of cytoskeletal/mitochondrial dynamics *Correspondence: and neurite outgrowth. In adult primary neurons, overexpression of M1R caused Mohammad G. Sabbir disruption of the tubulin, but not actin, cytoskeleton and significantly reduced neurite [email protected] outgrowth. Over-expression of a M1R-DREADD mutant comparatively increased neurite Specialty section: outgrowth suggesting that acetylcholine released from cultured neurons interacts This article was submitted to with M1R to suppress neurite outgrowth. -
WO 2017/177262 Al 19 October 2017 (19.10.2017) P O P C T
(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/177262 Al 19 October 2017 (19.10.2017) P O P C T (51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, A61K 31/198 (2006.01) A61P 27/02 (2006.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, A61K 31/375 (2006.01) A61P 27/10 (2006.01) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, (21) International Application Number: KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, PCT/AU20 17/0503 10 MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, (22) International Filing Date: NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, 10 April 2017 (10.04.2017) RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, (25) Filing Language: English ZA, ZM, ZW. (26) Publication Language: English (84) Designated States (unless otherwise indicated, for every (30) Priority Data: kind of regional protection available): ARIPO (BW, GH, 2016901339 11 April 2016 ( 11.04.2016) AU GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (71) Applicant: UNIVERSITY OF CANBERRA [AU/AU]; TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, University Drive, Bruce, Australian Capital Territory 2617 DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, (AU). -
Scorpion Toxins Targeted Against the Sarcoplasmic Reticulum Ca2+-Release Channel of Skeletal and Cardiac Muscle
Proc. Nati. Acad. Sci. USA Vol. 89, pp. 12185-12189, December 1992 Physiology Scorpion toxins targeted against the sarcoplasmic reticulum Ca2+-release channel of skeletal and cardiac muscle (ryanodine receptors/Pandinus imperator venom/planar bilayer/ventricular myocytes/Ca2+ indicator) HECTOR H. VALDIVIA*t, MARK S. KIRBYf, W. JONATHAN LEDERER*, AND ROBERTO CORONADO*t *Department of Physiology, University of Wisconsin School of Medicine, Madison, WI 53706; and tDepartment of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201 Communicated by Michael V. L. Bennett, September 21, 1992 ABSTRACT We report the purification of two peptides, peratoxin inhibitor (IpTxi)] or activated [imperatoxin activa- called "imperatoxin inhibitor" and "imperatoxin activator," tor (IpTxa)] ryanodine receptors of skeletal and cardiac from the venom of the scorpion Pandinus imperator targeted muscle. Part of these results have been communicated in an against ryanodine receptor Ca2+-release channels. Impera- abstract form (8). toxin inhibitor has a Mr of 10,500, inhibits [3Hjryanodine binding to skeletal and cardiac sarcoplasmic reticulum with an EDso of 10 nM, and blocks openings of skeletal and cardiac EXPERIMENTAL PROCEDURES Ca2+-release channels incorporated into planar bilayers. In Purification of Scorpion Toxins. Lyophilized P. imperator whole-cell recordings of cardiac myocytes, imperatoxin inhib- venom was obtained from Latoxan (Rosans, France). Venom itor decreased twitch amplitude and intracellular Ca2+ tran- (50 mg per batch) was extracted in 2-3 ml of deionized water sients, suggesting a selective blockade of Ca2+ release from the and chromatographed on a column (1. 5 x 125 cm) of Sepha- sarcoplasmic reticulum. Imperatoxin activator has a Mr of dex G-50 fine. -
ANA CAROLINA MARTINS WILLE.Pdf
UNIVERSIDADE FEDERAL DO PARANÁ ANA CAROLINA MARTINS WILLE AVALIAÇÃO DA ATIVIDADE DE FOSFOLIPASE-D RECOMBINANTE DO VENENO DA ARANHA MARROM (Loxosceles intermedia) SOBRE A PROLIFERAÇÃO, INFLUXO DE CÁLCIO E METABOLISMO DE FOSFOLIPÍDIOS EM CÉLULAS TUMORAIS. CURITIBA 2014 i Wille, Ana Carolina Martins Avaliação da atividade de fosfolipase-D recombinante do veneno da aranha marrom (Loxosceles intermedia) sobre a proliferação, influxo de cálcio e metabolismo de fosfolipídios em células tumorais Curitiba, 2014. 217p. Tese (Doutorado) – Universidade Federal do Paraná – UFPR 1.veneno de aranha marrom. 2. fosfolipase-D. 3.proliferação celular. 4.metabolismo de lipídios. 5.influxo de cálcio. ANA CAROLINA MARTINS WILLE AVALIAÇÃO DA ATIVIDADE DE FOSFOLIPASE-D RECOMBINANTE DO VENENO DA ARANHA MARROM (Loxosceles intermedia) SOBRE A PROLIFERAÇÃO, INFLUXO DE CÁLCIO E METABOLISMO DE FOSFOLIPÍDIOS EM CÉLULAS TUMORAIS. Tese apresentada como requisito à obtenção do grau de Doutor em Biologia Celular e Molecular, Curso de Pós- Graduação em Biologia Celular e Molecular, Setor de Ciências Biológicas, Universidade Federal do Paraná. Orientador(a): Dra. Andrea Senff Ribeiro Co-orientador: Dr. Silvio Sanches Veiga CURITIBA 2014 ii O desenvolvimento deste trabalho foi possível devido ao apoio financeiro do Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), a Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação Araucária e SETI-PR. iii Dedico este trabalho àquela que antes da sua existência foi o grande sonho que motivou minha vida. Sonho que foi a base para que eu escolhesse uma profissão e um trabalho. À você, minha amada filha GIOVANNA, hoje minha realidade, dedico todo meu trabalho. iv Dedico também este trabalho ao meu amado marido, amigo, professor e co- orientador Dr. -
Multiple Actions of Φ-LITX-Lw1a on Ryanodine Receptors Reveal a Functional Link Between Scorpion DDH and ICK Toxins
Multiple actions of φ-LITX-Lw1a on ryanodine receptors reveal a functional link between scorpion DDH and ICK toxins Jennifer J. Smitha, Irina Vettera, Richard J. Lewisa, Steve Peigneurb, Jan Tytgatb, Alexander Lamc, Esther M. Gallantc, Nicole A. Beardd, Paul F. Alewooda,1,2, and Angela F. Dulhuntyc,1,2 aChemical and Structural Biology, Institute for Molecular Biosciences, University of Queensland, St. Lucia, QLD 4072, Australia; bLaboratory of Toxicology, University of Leuven, 3000 Leuven, Belgium; cDepartment of Molecular Bioscience, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia; and dDiscipline of Biomedical Sciences, Faculty of Education, Science, Technology and Mathematics, University of Canberra, Canberra, ACT 2601, Australia Edited by Clara Franzini-Armstrong, University of Pennsylvania Medical Center, Philadelphia, PA, and approved April 23, 2013 (received for review August 26, 2012) We recently reported the isolation of a scorpion toxin named U1- mutations, atypical posttranslational modifications of RyRs, liotoxin-Lw1a (U1-LITX-Lw1a) that adopts an unusual 3D fold termed including hyperphosphorylation and S-nitrosylation, have been the disulfide-directed hairpin (DDH) motif, which is the proposed implicated in heart failure and pathologic muscle fatigue (10– evolutionary structural precursor of the three-disulfide-containing 12). Because of their wide distribution, there is mounting evidence inhibitor cystine knot (ICK) motif found widely in animals and plants. to suggest that dysregulation of RyRs also plays a role in a plethora Here we reveal that U1-LITX-Lw1a targets and activates the mam- of other disease states, including chronic pain, neurodegenerative malian ryanodine receptor intracellular calcium release channel (RyR) disorders such as Alzheimer’s disease, and intellectual deficit (13– with high (fM) potency and provides a functional link between DDH 15). -
Mechanistic Insights Into Functional Characteristics of Native Crotamine
Toxicon 146 (2018) 1e12 Contents lists available at ScienceDirect Toxicon journal homepage: www.elsevier.com/locate/toxicon Mechanistic insights into functional characteristics of native crotamine Daniel Batista da Cunha a, Ana Vitoria Pupo Silvestrini a, Ana Carolina Gomes da Silva a, Deborah Maria de Paula Estevam b,Flavia Lino Pollettini b, Juliana de Oliveira Navarro a, Armindo Antonio^ Alves a, Ana Laura Remedio Zeni Beretta a, * Joyce M. Annichino Bizzacchi c, Lilian Cristina Pereira d, Maurício Ventura Mazzi a, a Graduate Program in Biomedical Sciences Hermínio Ometto University Center, UNIARARAS, 7 Av. Dr. Maximiliano Baruto, 500, CEP 13607-339, Araras, SP, Brazil b Graduate Program in Agrarian and Veterinary Sciences, State University Paulista Júlio de Mesquita Filho-UNESP, Jaboticabal, SP, Brazil c Blood Hemostasis Laboratory, Faculty of Medical Sciences, State University of Campinas, Campinas, SP, Brazil d Department of Bioprocesses and Biotechnology, Faculty of Agronomic Sciences, State University Paulista Júlio Mesquita Filho-UNESP, Botucatu, SP, Brazil article info abstract Article history: The chemical composition of snake venoms is a complex mixture of proteins and peptides that can be Received 4 October 2017 pharmacologically active. Crotamine, a cell-penetrating peptide, has been described to have antimicro- Received in revised form bial properties and it exerts its effects by interacting selectively with different structures, inducing 6 March 2018 changes in the ion flow pattern and cellular responses. However, its real therapeutic potential is not yet Accepted 20 March 2018 fully known. Bearing in mind that crotamine is a promising molecule in therapeutics, this study inves- Available online 21 March 2018 tigated the action of purified molecule in three aspects: I) antibacterial action on different species of clinical interest, II) the effect of two different concentrations of the molecule on platelet aggregation, and Keywords: fi Crotalus durissus terrificus III) its effects on isolated mitochondria. -
Snakes: Cultural Beliefs and Practices Related to Snakebites in a Brazilian Rural Settlement Dídac S Fita1, Eraldo M Costa Neto2*, Alexandre Schiavetti3
Fita et al. Journal of Ethnobiology and Ethnomedicine 2010, 6:13 http://www.ethnobiomed.com/content/6/1/13 JOURNAL OF ETHNOBIOLOGY AND ETHNOMEDICINE RESEARCH Open Access ’Offensive’ snakes: cultural beliefs and practices related to snakebites in a Brazilian rural settlement Dídac S Fita1, Eraldo M Costa Neto2*, Alexandre Schiavetti3 Abstract This paper records the meaning of the term ‘offense’ and the folk knowledge related to local beliefs and practices of folk medicine that prevent and treat snake bites, as well as the implications for the conservation of snakes in the county of Pedra Branca, Bahia State, Brazil. The data was recorded from September to November 2006 by means of open-ended interviews performed with 74 individuals of both genders, whose ages ranged from 4 to 89 years old. The results show that the local terms biting, stinging and pricking are synonymous and used as equivalent to offending. All these terms mean to attack. A total of 23 types of ‘snakes’ were recorded, based on their local names. Four of them are Viperidae, which were considered the most dangerous to humans, besides causing more aversion and fear in the population. In general, local people have strong negative behavior towards snakes, killing them whenever possible. Until the antivenom was present and available, the locals used only charms, prayers and homemade remedies to treat or protect themselves and others from snake bites. Nowadays, people do not pay attention to these things because, basically, the antivenom is now easily obtained at regional hospitals. It is under- stood that the ethnozoological knowledge, customs and popular practices of the Pedra Branca inhabitants result in a valuable cultural resource which should be considered in every discussion regarding public health, sanitation and practices of traditional medicine, as well as in faunistic studies and conservation strategies for local biological diversity. -
Paula Henriques Cruz Ciscotto
PAULA HENRIQUES CRUZ CISCOTTO Purificação e caracterização biológica (estrutural, antibacteriana, antiparasitária, hemolítica e antigênica) de componentes do veneno da serpente Bothrops jararaca. BELO HORIZONTE MINAS GERAIS-BRASIL 2005 PAULA HENRIQUES CRUZ CISCOTTO Purificação e caracterização biológica (estrutural, antibacteriana, antiparasitária, hemolítica e antigênica) de componentes do veneno da serpente Bothrops jararaca. Dissertação submetida ao Curso de Pós- graduação em Bioquímica e Imunologia do Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais como requisito parcial para obtenção do título de mestre em Bioquímica e Imunologia. Orientador: Prof. Dr. Carlos Chávez Olórtegui BELO HORIZONTE MINAS GERAIS-BRASIL 2005 Este trabalho foi desenvolvido no Laboratório de Imunoquímica de Toxinas, do Departamento de Bioquímica e Imunologia do Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais. Contando com a colaboração do Laboratório de Microbiologia Oral e Anaeróbios, do Departamento de Microbiologia, do Laboratório de Enzimologia e Físico- Química de Proteínas, do Núcleo de Estrutura e Função de Biomoléculas, ambos do Departamento de Bioquímica e Imunologia, todos do Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais. ii Agradecimentos Aos meus pais por sempre estarem ao meu lado, incentivando meu crescimento pessoal e profissional, e apoiando minhas decisões. Ao Luiz pelo carinho constante e incentivo permanente, por sua tolerância em momentos delicados. Graças à suas palavras de perseverança e otimismo consegui chegar ao final desta caminhada. Ao Prof. Carlos Chávez Olórtegui por me incentivar a começar esta trajetória científica pelo caminho da especialização profissional e por depositar em mim sua confiança para realização deste trabalho. Ao Jamil por seus ensinamentos, incentivo e por estar sempre disponível a me ajudar. -
(12) United States Patent (10) Patent No.: US 9,062,119 B2 Varga Et Al
USOO90621-19B2 (12) United States Patent (10) Patent No.: US 9,062,119 B2 Varga et al. (45) Date of Patent: Jun. 23, 2015 (54) MODIFIED PEPTIDE TOXINS OTHER PUBLICATIONS (71) Applicants:Zoltan Varga, Debrecen (HU); Gyorgy Bagdanyi, M. et al. Anuroctoxin, a new scorpion toxin of the alpha Panyi, Debrecen (HU); Gabor Toth, KTX 6 subfamily, is highly selective for Kv1.3 over IKCal ion Szeged (HU); Kinga Rakosi, Szeged channels of human T lymphocytes. Mol Pharmacol (2005), vol. 67. (HU) pp. 1034-1044. Batista, CV. et al. Two novel toxins from the Amazonian scorpion (72) Inventors: Zoltan Varga, Debrecen (HU); Gyorgy Tityus cambridgei that block Kv1.3 and Shaker BK(+)-channels with Panyi, Debrecen (HU); Gabor Toth, distinctly different affinities. Biochim Biophys Acta (2002), vol. Szeged (HU); Kinga Rakosi, Szeged 1601, pp. 123-131. Beeton, C. et al. Selective blockade of T lymphocyte K+ channels (HU) ameliorates experimental autoimmune encephalomyelitis, a model (73) Assignees: University of Debrecen, Debrecen for multiple sclerosis. Proc Natl Acad Sci USA (2001), vol. 98, pp. (HU); University of Szeged, Szeged 13942-13947. Beeton, C. et al. Kv1.3 channels are a therapeutic target for T cell (HU) mediated autoimmune diseases. Proc Natl AcadSci USA (2006), vol. 103, pp. 17414-17419. (*) Notice: Subject to any disclaimer, the term of this Corzo, G. et al. A selective blocker of Kv1.2 and Kv1.3 potassium patent is extended or adjusted under 35 channels from the venom of the Scorpion Centruroides suffusus suf U.S.C. 154(b) by 0 days. fusus. Biochem Pharmacol (2008), vol. -
Maurocalcine-Derivatives As Biotechnological Tools for The
Maurocalcine-derivatives as biotechnological tools for the penetration of cell-impermeable compounds Narendra Ram, Emilie Jaumain, Michel Ronjat, Fabienne Pirollet, Michel de Waard To cite this version: Narendra Ram, Emilie Jaumain, Michel Ronjat, Fabienne Pirollet, Michel de Waard. Maurocalcine- derivatives as biotechnological tools for the penetration of cell-impermeable compounds: Technological value of a scorpion toxin. de Lima, M.E.; de Castro Pimenta, A.M.; Martin-Eauclaire, M.F.; Bendeta Zengali, R.; Rochat, H. E. Animal Toxins: state of the art: Perspectives in Health and Biotechnology., Editora UFMG Belo Horizonte, Brazil, pp.715-732, 2009. inserm-00515224 HAL Id: inserm-00515224 https://www.hal.inserm.fr/inserm-00515224 Submitted on 5 Sep 2013 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Maurocalcine-derivatives as biotechnological tools for the penetration of cell-impermeable compounds by Narendra Ram1,2, Emilie Jaumain1,2, Michel Ronjat1,2, Fabienne Pirollet1,2 & Michel De Waard1,2* 1 INSERM, U836, Team 3: Calcium channels, functions and pathologies, BP 170, Grenoble Cedex 9, F-38042, France 2 Université Joseph Fourier, Institut des Neurosciences, BP 170, Grenoble Cedex 9, F-38042, France Running title: Technological value of a scorpion toxin *To whom correspondence should be sent: Dr.