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1.
Curr Biol ; 30(21): 4316-4321.e2, 2020 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-32916114

RESUMO

Arachnids are the second most successful terrestrial animal group after insects [1] and were one of the first arthropod clades to successfully invade land [2]. Fossil evidence for this transition is limited, with the majority of arachnid clades first appearing in the terrestrial fossil record. Furthermore, molecular clock dating has suggested a Cambrian-Ordovician terrestrialization event for arachnids [3], some 60 Ma before their first fossils in the Silurian, although these estimates assume that arachnids evolved from a fully aquatic ancestor. Eurypterids, the sister clade to terrestrial arachnids [4-6], are known to have undergone major macroecological shifts in transitioning from marine to freshwater environments during the Devonian [7, 8]. Discoveries of apparently subaerial eurypterid trackways [9, 10] have led to the suggestion that eurypterids were even able to venture on land and possibly breathe air [11]. However, modern horseshoe crabs undertake amphibious excursions onto land to reproduce [12], rendering trace fossil evidence alone inconclusive. Here, we present details of the respiratory organs of Adelophthalmus pyrrhae sp. nov. from the Carboniferous of Montagne Noire, France [13], revealed through micro computed tomography (µ-CT) imaging. Pillar-like trabeculae on the dorsal surface of each gill lamella indicate eurypterids were capable of subaerial breathing, suggesting that book gills are the direct precursors to book lungs while vascular ancillary respiratory structures known as Kiemenplatten represent novel air-breathing structures. The discovery of air-breathing structures in eurypterids indicates that characters permitting terrestrialization accrued in the arachnid stem lineage and suggests the Cambrian-Ordovician ancestor of arachnids would also have been semi-terrestrial.


Assuntos
Evolução Biológica , Respiração , Sistema Respiratório/anatomia & histologia , Escorpiões/fisiologia , Animais , Organismos Aquáticos/fisiologia , Fósseis/anatomia & histologia , Fósseis/diagnóstico por imagem , Caranguejos Ferradura/anatomia & histologia , Caranguejos Ferradura/fisiologia , Sistema Respiratório/diagnóstico por imagem , Escorpiões/anatomia & histologia , Microtomografia por Raio-X
2.
Toxicon ; 185: 36-45, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32585220

RESUMO

Scorpionism is a serious public health problem in various regions of the world. In Brazil, a high number of accidents by scorpions have been reported. From 2014 to 2018, about 547,000 cases were recorded, resulting in 466 deaths. The scorpion Tityus stigmurus is the predominant species in the northeast of Brazil, being responsible for most scorpionism cases in this region. With the aid of the transcriptomic approach of the venom gland of this species, components as neurotoxins, antimicrobials, metal chelating peptides and hypotensins, have been identified and characterized in silico, showing different biologic activity in vitro. In addition, the neuronal, pancreatic, renal, and enzymatic effects have been demonstrated for the crude T. stigmurus venom. Therefore, the T. stigmurus scorpion venom constitutes a rich arsenal of bioactive molecules with high potential for therapeutic and biotechnological application.


Assuntos
Picadas de Escorpião , Venenos de Escorpião/toxicidade , Escorpiões/fisiologia , Animais , Brasil , Rim , Neurotoxinas , Peptídeos
3.
Molecules ; 24(14)2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-31340554

RESUMO

Scorpions, a characteristic group of arthropods, are among the earliest diverging arachnids, dating back almost 440 million years. One of the many interesting aspects of scorpions is that they have venom arsenals for capturing prey and defending against predators, which may play a critical role in their evolutionary success. Unfortunately, however, scorpion envenomation represents a serious health problem in several countries, including Iran. Iran is acknowledged as an area with a high richness of scorpion species and families. The diversity of the scorpion fauna in Iran is the subject of this review, in which we report a total of 78 species and subspecies in 19 genera and four families. We also list some of the toxins or genes studied from five species, including Androctonus crassicauda, Hottentotta zagrosensis, Mesobuthus phillipsi, Odontobuthus doriae, and Hemiscorpius lepturus, in the Buthidae and Hemiscorpiidae families. Lastly, we review the diverse functions of typical toxins from the Iranian scorpion species, including their medical applications.


Assuntos
Peptídeos Catiônicos Antimicrobianos/química , Antineoplásicos/química , Proteínas de Artrópodes/química , Venenos de Escorpião/química , Escorpiões/química , Animais , Peptídeos Catiônicos Antimicrobianos/biossíntese , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Proteínas de Artrópodes/biossíntese , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/uso terapêutico , Descoberta de Drogas/métodos , Expressão Gênica , Humanos , Canais Iônicos/agonistas , Canais Iônicos/antagonistas & inibidores , Canais Iônicos/metabolismo , Irã (Geográfico) , Metaloproteases/biossíntese , Metaloproteases/isolamento & purificação , Metaloproteases/toxicidade , Fosfolipases A2/biossíntese , Fosfolipases A2/isolamento & purificação , Fosfolipases A2/toxicidade , Filogenia , Picadas de Escorpião/fisiopatologia , Venenos de Escorpião/biossíntese , Venenos de Escorpião/isolamento & purificação , Escorpiões/classificação , Escorpiões/patogenicidade , Escorpiões/fisiologia , Inibidores de Serina Proteinase/biossíntese , Inibidores de Serina Proteinase/isolamento & purificação , Inibidores de Serina Proteinase/toxicidade , Especificidade da Espécie
4.
J Zoo Wildl Med ; 50(1): 111-122, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31120669

RESUMO

Large arachnids are commonly managed under professional care, and anesthesia is occasionally required for physical examination and diagnostic and therapeutic procedures. Anesthetic responses and hemolymph gas analysis have been studied previously in spiders, but scorpions have yet to be investigated. This study measured hemolymph gas values with an i-STAT point of care blood gas analyzer in healthy adult Asian forest scorpions (Heterometrus longimanus = HL, n = 8) and dictator scorpions (Pandinus dictator = PD, n = 12) breathing: 1) room air (RA), 2) 100% oxygen for 10 min in a chamber (OX), and 3) 5% isoflurane and oxygen (ISO) in a chamber until induction or loss of righting reflex. All scorpions recovered without complications, and there were no cartridge failures. Analysis of hemolymph gas values revealed that pH was lower in OX compared with RA and ISO and was lower in PD compared with HL scorpions. The partial pressure of carbon dioxide did not differ between inhaled gases but was higher in PD compared with HL. The partial pressure of oxygen (pO2) was higher in ISO compared with OX, and both were higher than when breathing RA. Despite a lack of species difference in pO2, PD had a more dramatic increase in pO2 in ISO compared with HL (significant species and inhalant interaction). PD had a significantly shorter induction time than HL, but recovery time (return of righting reflex) did not differ between species. Subjectively, HL exhibited rough inductions compared with PD, characterized by violent whole-body and tail movements. The unexpected increase in pO2 in ISO compared with OX, along with the species-specific differences and anesthetic effects, emphasizes the unique respiratory physiology of scorpions and demonstrates that further species-specific studies of anesthetics are warranted.


Assuntos
Anestésicos Inalatórios/efeitos adversos , Isoflurano/efeitos adversos , Oxigênio/efeitos adversos , Escorpiões/efeitos dos fármacos , Aranhas/efeitos dos fármacos , Anestésicos Inalatórios/administração & dosagem , Animais , Gasometria/veterinária , Hemolinfa/química , Isoflurano/administração & dosagem , Oxigênio/administração & dosagem , Reflexo de Endireitamento/efeitos dos fármacos , Escorpiões/fisiologia , Aranhas/fisiologia
5.
J Cell Mol Med ; 22(9): 4460-4473, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29993185

RESUMO

In this study, a novel scorpion venom-derived peptide named Gonearrestide was identified in an in-house constructed scorpion venom library through a combination of high-throughput NGS transcriptome and MS/MS proteome platform. In total, 238 novel peptides were discovered from two scorpion species; and 22 peptides were selected for further study after a battery of functional prediction analysis. Following a series of bioinformatics analysis alongside with in vitro biological functional screenings, Gonearrestide was found to be a highly potent anticancer peptide which acts on a broad spectrum of human cancer cells while causing few if any observed cytotoxic effects on epithelial cells and erythrocytes. We further investigated the precise anticancer mechanism of Gonearrestide by focusing on its effects on the colorectal cancer cell line, HCT116. NGS RNA sequencing was employed to obtain full gene expression profiles in HCT116 cells, cultured in the presence and absence of Gonearrestide, to dissect signalling pathway differences. Taken together the in vitro, in vivo and ex vivo validation studies, it was proven that Gonearrestide could inhibit the growth of primary colon cancer cells and solid tumours by triggering cell cycle arrest in G1 phase through inhibition of cyclin-dependent kinases 4 (CDK4) and up-regulate the expression of cell cycle regulators/inhibitors-cyclin D3, p27, and p21. Furthermore, prediction of signalling pathways and potential binding sites used by Gonearrestide are also presented in this study.


Assuntos
Antineoplásicos/farmacologia , Neoplasias do Colo/tratamento farmacológico , Fase G1/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Peptídeos/farmacologia , Venenos de Escorpião/química , Animais , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Sítios de Ligação , Linhagem Celular Tumoral , Neoplasias do Colo/genética , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Ciclina D3/genética , Ciclina D3/metabolismo , Quinase 4 Dependente de Ciclina/genética , Quinase 4 Dependente de Ciclina/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Feminino , Fase G1/genética , Células HCT116 , Humanos , Camundongos Nus , Peptídeos/química , Peptídeos/isolamento & purificação , Ligação Proteica , Escorpiões/fisiologia , Transdução de Sinais , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Int Immunopharmacol ; 61: 256-265, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29902709

RESUMO

Antivenom treatment has been largely used against scorpion stings. Despite their efficacy, the use of mammalian antivenoms may cause adverse effects due to the immune system activation. IgYs from hyperimmunized laying hens against venoms could be a promising alternative to equine IgGs due to the various benefits that these antibodies can provide. Here we report the preparation of specific IgYs by immunizing laying hens with Aah (Androctonus australis hector) scorpion venom. IgYs were isolated from egg yolks by water dilution and salt precipitation methods; they were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis, western blot and ELISA. The efficiency of these immunoglobulins on the pathophysiological effects induced by Aah venom was assessed by histological and metabolical analysis of the aorta and the heart. The inflammatory response was assessed by evaluating the granulocyte tissue infiltration and oxidative/nitrosative status. Results revealed high IgYs titers against Aah venom by ELISA. Overall, these IgYs seem to protect efficiently mice against envenomation and neutralized the lethal effects of scorpion venom with a high efficacy; the median effective dose (ED50) was 221 µl/2 LD50; i.e. an amount of 79.23 mg of IgY scan neutralize 1 mg of Aah venom. IgY antibodies neutralize effectively the Aah venom lethality and could prevent severe pathological effects induced by scorpion venom and could be used as an effective alternative to equine IgGs against scorpion envenoming.


Assuntos
Aorta/patologia , Galinhas/imunologia , Imunoglobulinas/uso terapêutico , Imunoterapia Adotiva/métodos , Miocárdio/patologia , Picadas de Escorpião/terapia , Venenos de Escorpião/imunologia , Animais , Antivenenos/uso terapêutico , Humanos , Imunização , Masculino , Camundongos , Camundongos Endogâmicos , Picadas de Escorpião/imunologia , Escorpiões/fisiologia
7.
Sci Rep ; 7: 44685, 2017 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-28322221

RESUMO

Complementary and alternative medicine (CAM) is the term used to describe many kinds of products, practices, and systems that are not part of conventional medicine. Cancer patients usually do everything they can to combat the disease, manage its symptoms, and cope with the side effects of treatment. Unfortunately, patients who use CAM underestimate the risk of interaction with cancer therapy or worse they omit conventional therapy thus reducing the possibility of cancer remission. Herein we analyzed the effects of Vidatox 30 CH (venom extracted from the Junceus Rhopalurus scorpion) on hepatocellular carcinoma (HCC), the second leading cause of cancer-related deaths. We found out that Vidatox increases HCC proliferation and invasion whereas it does not seem to interact with sorafenib, the orally active multikinase inhibitor approved for the treatment of advanced hepatocellular carcinoma. Our results suggest that the concentration of Vidatox used in the present study has not anti-neoplastic effects and care must be taken in hiring Vidatox in patients with HCC.


Assuntos
Antineoplásicos/farmacologia , Carcinoma Hepatocelular/induzido quimicamente , Regulação Neoplásica da Expressão Gênica , Hepatócitos/efeitos dos fármacos , Neoplasias Hepáticas/induzido quimicamente , Niacinamida/análogos & derivados , Compostos de Fenilureia/farmacologia , Venenos de Aranha/toxicidade , Animais , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citocinas/genética , Citocinas/metabolismo , Dietilnitrosamina , Células Hep G2 , Hepatócitos/metabolismo , Hepatócitos/patologia , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Masculino , Niacinamida/farmacologia , Ratos Wistar , Escorpiões/química , Escorpiões/patogenicidade , Escorpiões/fisiologia , Transdução de Sinais , Sorafenibe , Venenos de Aranha/antagonistas & inibidores , Venenos de Aranha/isolamento & purificação , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
8.
Rev. biol. trop ; 64(3): 1019-1027, jul.-sep. 2016. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-958192

RESUMO

Abstract:Approximately 2 000 scorpion species can be found around the world; although few species are considered "harmful" to human beings, a high number of scorpionism cases are reported all over the world. The elaboration of anti-scorpion sera requires the establishment of an animal collection maintained in captivity for venom extraction purposes. The Clodomiro Picado Institute (ICP, for its acronym in Spanish), poses a vast trajectory in manufacturing snakebite antivenoms, and starts a scorpion collection in 2005 for this purpose. In total, 2 043 scorpions were classified in 11 species and collected during a seven-year period using a black-light flashlight and an intensive seeking methodology. The scorpions were collected from several localities of the Pacific and the Caribbean versants of Costa Rica. The venom extraction was performed by applying electrostimulation; the collected venom was characterized by total protein content in addition to median lethal doses. Centruroidesbicolor showed higher amounts of venom yield, total protein content and more lethal dose, all of which were correlated with its body mass. The techniques used to keep scorpions in captivity allowed the animals to live several years. Longevity analysis showed significant differences among scorpion genera (H= 353.80; df= 3; P < 0.0001); moreover, the genus Didymocentrus lived longer with an average of 4.46 years. One key factor of its longevity was that it did not go through venom extraction processes. Additionally, a high survival rate of Tityuspachyurus born in captivity, compared to other species within the same genus, was observed (H= 94.32; df= 3; P < 0.0001). This characteristic should be taken into consideration, when programs of reproduction in captivity are designed. In conclusion, the maintenance of a scorpion collection was efficient for venom extraction purposes and a longer life expectancy of the animals. Moreover, there is a scarcity on publications regarding scorpion maintenance in captivity for venom extraction purposes; therefore, a deeper research in aspects such as reproduction, death causes and feeding behaviors is required. Rev. Biol. Trop. 64 (3): 1019-1027. Epub 2016 September 01.


ResumenExisten alrededor de 2 000 especies de escorpiones en el mundo y a pesar de que muy pocas de ellas son consideradas peligrosas para el ser humano se presentan muchos casos de escorpionismo alrededor del mundo. La elaboración de suero antiescorpiónico requiere en primera instancia del establecimiento de una colección en cautiverio para la extracción del veneno, razón por la cual el Instituto Clodomiro Picado (ICP), con una amplia trayectoria en la elaboración de suero antiofídico, inicia en el 2005 una colección para dicho fin. En total 2 043 escorpiones clasificados en 11 especies fueron recolectados en un periodo de siete años mediante búsqueda intensiva con luz ultravioleta en varias localidades de la vertiente Pacífica, así como la vertiente Atlántica de Costa Rica. Las extracciones de veneno se realizaron con electro-estimulación, y el veneno recolectado fue caracterizado por el contenido total de proteínas, además se realizaron pruebas de letalidad, siendo Centruroides bicolor el más letal y el que presento los valores más altos en cantidad de veneno y proteínas totales lo cual es consecuente con su masa corporal. Las técnicas empleadas en el mantenimiento en cautiverio de los escorpiones permitieron que los escorpiones vivieran varios años, en un análisis de la longevidad se obtuvieron diferencias significativas entre los géneros (H = 353.80; g.l = 3; P < 0.0001), siendo el género Didymocentrus el más longevo con un promedio de 4.46 años, un factor importante de su longevidad es el hecho de no haber formado parte del proceso de extracción. También se pudo evidenciar una alta supervivencia de crías nacidas en cautiverio de la especie Tityus pachyurus con respecto a las otras especies del mismo género (H = 94.32; g.l = 3; P < 0.0001), resultado que puede ser tomado en cuenta para reproducir esta especie en cautiverio. El mantenimiento de la colección de escorpiones fue eficiente para el proceso de extracción de veneno y para una supervivencia prolongada de los animales. Existen muy pocas publicaciones en lo que se refiere al mantenimiento de escorpiones en cautiverio para la extracción de veneno por lo que se requiere de una investigación más profunda en aspectos como la reproducción, las causas de muerte y la alimentación.


Assuntos
Animais , Venenos de Escorpião/isolamento & purificação , Escorpiões/fisiologia , Antivenenos , Especificidade da Espécie , Fatores de Tempo , Expectativa de Vida , Costa Rica , Distribuição Animal , Longevidade
9.
J Biochem Mol Toxicol ; 30(12): 580-587, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27271987

RESUMO

Kaliotoxin (KTX), a specific blocker of potassium channels, exerts various toxic effects due to its action on the central nervous system. Its use in experimental model could help the understanding of the cellular and molecular mechanisms involved in the neuropathological processes related to potassium channel dysfunctions. In this study, the ability of KTX to stimulate neuro-immuno-endocrine axis was investigated. As results, the intracerebroventricular injection of KTX leads to severe structural-functional alterations of both hypothalamus and thyroid. These alterations were characterized by a massive release of hormones' markers of thyroid function associated with damaged tissue which was infiltrated by inflammatory cell and an imbalanced redox status. Taken together, these data highlight that KTX is able to modulate the neuro-endocrine response after binding to its targets leading to the hypothalamus and the thyroid stimulation, probably by inflammatory response activation and the installation of oxidative stress in these organs.


Assuntos
Eosinófilos/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Venenos de Escorpião/toxicidade , Escorpiões/química , Glândula Tireoide/efeitos dos fármacos , Animais , Calcitonina/biossíntese , Calcitonina/metabolismo , Catalase/metabolismo , Eosinófilos/imunologia , Glutationa/metabolismo , Hipotálamo/imunologia , Hipotálamo/metabolismo , Injeções Intraventriculares , Malondialdeído/metabolismo , Camundongos , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Nitrilas/metabolismo , Oxirredução , Estresse Oxidativo , Venenos de Escorpião/isolamento & purificação , Escorpiões/fisiologia , Glândula Tireoide/imunologia , Glândula Tireoide/metabolismo , Tireotropina/biossíntese , Tireotropina/metabolismo , Tiroxina/biossíntese , Tiroxina/metabolismo , Tri-Iodotironina/biossíntese , Tri-Iodotironina/metabolismo
10.
J Biol Chem ; 291(13): 7097-106, 2016 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-26817841

RESUMO

The structural similarity between defensins and scorpion neurotoxins suggests that they might have evolved from a common ancestor. However, there is no direct experimental evidence demonstrating a functional link between scorpion neurotoxins and defensins. The scorpion defensin BmKDfsin4 from Mesobuthus martensiiKarsch contains 37 amino acid residues and a conserved cystine-stabilized α/ß structural fold. The recombinant BmKDfsin4, a classical defensin, has been found to have inhibitory activity against Gram-positive bacteria such as Staphylococcus aureus, Bacillus subtilis, and Micrococcus luteusas well as methicillin-resistant Staphylococcus aureus Interestingly, electrophysiological experiments showed that BmKDfsin4,like scorpion potassium channel neurotoxins, could effectively inhibit Kv1.1, Kv1.2, and Kv1.3 channel currents, and its IC50value for the Kv1.3 channel was 510.2 nm Similar to the structure-function relationships of classical scorpion potassium channel-blocking toxins, basic residues (Lys-13 and Arg-19) of BmKDfsin4 play critical roles in peptide-Kv1.3 channel interactions. Furthermore, mutagenesis and electrophysiological experiments demonstrated that the channel extracellular pore region is the binding site of BmKDfsin4, indicating that BmKDfsin4 adopts the same mechanism for blocking potassium channel currents as classical scorpion toxins. Taken together, our work identifies scorpion BmKDfsin4 as the first invertebrate defensin to block potassium channels. These findings not only demonstrate that defensins from invertebrate animals are a novel type of potassium channel blockers but also provide evidence of a functional link between defensins and neurotoxins.


Assuntos
Antibacterianos/química , Defensinas/química , Neurotoxinas/química , Bloqueadores dos Canais de Potássio/química , Venenos de Escorpião/química , Sequência de Aminoácidos , Animais , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/crescimento & desenvolvimento , Defensinas/genética , Defensinas/metabolismo , Defensinas/farmacologia , Expressão Gênica , Humanos , Canal de Potássio Kv1.1/antagonistas & inibidores , Canal de Potássio Kv1.1/genética , Canal de Potássio Kv1.1/metabolismo , Canal de Potássio Kv1.2/antagonistas & inibidores , Canal de Potássio Kv1.2/genética , Canal de Potássio Kv1.2/metabolismo , Canal de Potássio Kv1.3/antagonistas & inibidores , Canal de Potássio Kv1.3/genética , Canal de Potássio Kv1.3/metabolismo , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Camundongos , Micrococcus luteus/efeitos dos fármacos , Micrococcus luteus/crescimento & desenvolvimento , Modelos Moleculares , Dados de Sequência Molecular , Neurotoxinas/genética , Neurotoxinas/metabolismo , Neurotoxinas/farmacologia , Bloqueadores dos Canais de Potássio/metabolismo , Bloqueadores dos Canais de Potássio/farmacologia , Domínios e Motivos de Interação entre Proteínas , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Venenos de Escorpião/biossíntese , Escorpiões/química , Escorpiões/fisiologia , Alinhamento de Sequência , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Homologia Estrutural de Proteína , Relação Estrutura-Atividade
11.
J Pept Sci ; 21(8): 636-43, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25919411

RESUMO

La1 is a 73-residue cysteine-rich peptide isolated from the scorpion Liocheles australasiae venom. Although La1 is the most abundant peptide in the venom, its biological function remains unknown. Here, we describe a method for efficient chemical synthesis of La1 using the native chemical ligation (NCL) strategy, in which three peptide components of less than 40 residues were sequentially ligated. The peptide thioester necessary for NCL was synthesized using an aromatic N-acylurea approach with Fmoc-SPPS. After completion of sequential NCL, disulfide bond formation was carried out using a dialysis method, in which the linear peptide dissolved in an acidic solution was dialyzed against a slightly alkaline buffer to obtain correctly folded La1. Next, we determined the disulfide bonding pattern of La1. Enzymatic and chemical digests of La1 without reduction of disulfide bonds were analyzed by liquid chromatography/mass spectrometry (LC/MS), which revealed two of four disulfide bond linkages. The remaining two linkages were assigned based on MS/MS analysis of a peptide fragment containing two disulfide bonds. Consequently, the disulfide bonding pattern of La1 was found to be similar to that of a von Willebrand factor type C (VWC) domain. To our knowledge, this is the first report of the experimental determination of the disulfide bonding pattern of peptides having a single VWC domain as well as their chemical synthesis. La1 synthesized in this study will be useful for investigation of its biological role in the venom.


Assuntos
Proteínas de Artrópodes/síntese química , Venenos de Escorpião/química , Escorpiões/química , Sulfetos/química , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/isolamento & purificação , Estrutura Molecular , Venenos de Escorpião/síntese química , Escorpiões/fisiologia , Espectrometria de Massas em Tandem , Fator de von Willebrand/química
12.
Artigo em Inglês | LILACS | ID: lil-724678

RESUMO

Scorpion envenomations are a major public health problem in Brazil, whose most dangerous cases are attributable to the genus Tityus. This study was designed to compare the clinical and demographic features of envenomations by 77tyus obscurus in two areas of the state of Pará located in the Amazon basin. Were compared demographic findings, local and systemic signs and symptoms of human envenomations caused by T. obscurus that occurred in western and eastern areas of the state.


Assuntos
Animais , Intoxicação/complicações , Neurologia , Venenos/análise , Escorpiões/fisiologia
13.
J. venom. anim. toxins incl. trop. dis ; 20: 1-7, 04/02/2014. tab, ilus, map
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484567

RESUMO

Scorpion envenomations are a major public health problem in Brazil, whose most dangerous cases are attributable to the genus Tityus. This study was designed to compare the clinical and demographic features of envenomations by 77tyus obscurus in two areas of the state of Pará located in the Amazon basin. Were compared demographic findings, local and systemic signs and symptoms of human envenomations caused by T. obscurus that occurred in western and eastern areas of the state.


Assuntos
Animais , Intoxicação/complicações , Neurologia , Venenos/análise , Escorpiões/fisiologia
14.
Toxicon ; 63: 44-54, 2013 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-23182832

RESUMO

The Urodacidae scorpions are the most widely distributed of the four families in Australia and represent half of the species in the continent, yet their venoms remain largely unstudied. This communication reports the first results of a proteome analysis of the venom of the scorpion Urodacus yaschenkoi performed by mass fingerprinting, after high performance liquid chromatography (HPLC) separation. A total of 74 fractions were obtained by HPLC separation allowing the identification of approximately 274 different molecular masses with molecular weights varying from 287 to 43,437 Da. The most abundant peptides were those from 1 K Da and 4-5 K Da representing antimicrobial peptides and putative potassium channel toxins, respectively. Three such peptides were chemically synthesized and tested against Gram-positive and Gram-negative bacteria showing minimum inhibitory concentration in the low micromolar range, but with moderate hemolytic activity. It also reports a transcriptome analysis of the venom glands of the same scorpion species, undertaken by constructing a cDNA library and conducting random sequencing screening of the transcripts. From the resultant cDNA library 172 expressed sequence tags (ESTs) were analyzed. These transcripts were further clustered into 120 unique sequences (23 contigs and 97 singlets). The identified putative proteins can be assorted in several groups, such as those implicated in common cellular processes, putative neurotoxins and antimicrobial peptides. The scorpion U. yaschenkoi is not known to be dangerous to humans and its venom contains peptides similar to those of Opisthacanthus cayaporum (antibacterial), Scorpio maurus palmatus (maurocalcin), Opistophthalmus carinatus (opistoporines) and Hadrurus gerstchi (scorpine-like molecules), amongst others.


Assuntos
Antibacterianos/análise , DNA Complementar/química , Peptídeos/farmacologia , Venenos de Escorpião/química , Escorpiões/fisiologia , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Eritrócitos/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Hemolíticos/análise , Hemolíticos/isolamento & purificação , Hemolíticos/farmacologia , Humanos , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Peso Molecular , Mapeamento de Peptídeos , Peptídeos/química , Peptídeos/isolamento & purificação , Bloqueadores dos Canais de Potássio/análise , Bloqueadores dos Canais de Potássio/farmacologia
15.
Toxicon ; 63: 70-7, 2013 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-23186624

RESUMO

A study was conducted to assess the effect of clonidine (α(2)-adrenoceptor selective agonist) on glycemia, serum and urine α-amylase, blood urea nitrogen (BUN), serum creatinine, white blood cell count, kidney histology and zymogen granule content in pancreatic acini, in mice under the effect of Tityus discrepans (Td) scorpion venom. BALB/c male mice (20 ± 2 g, n = 7-11) were intraperitoneally (ip) injected with a sublethal dose (1 µg/g) of Td venom, and were treated (ip) with 0.1 µg/g of clonidine (Catapresan(®)) or 0.9% NaCl 30 min after the venom injection, and then every 2 h. Six hours later, mice were anesthetized with diethylether and urine and blood samples were withdrawn by cystocentesis and cardiocentesis, respectively. Tissue samples were obtained and fixed immediately in buffered formalin (2%, pH 7.4) and then processed for stain H&E. Td venom did not cause hyperglycemia by itself. However, clonidine induced hyperglycemia, which was synergized by Td venom. Although the venom did not produce hyperamylasemia, clonidine significantly diminished serum α-amylase activity in envenomed mice. Td venom did not significantly increase urinary α-amylase activity, which was unaffected by clonidine. Morphometric analysis using microphotographs of pancreata from mice injected with Td venom showed a reduced zymogen granule content as judged by the acidophilic bidimensional area of acini. This effect was significantly reduced by clonidine. Kidney samples showed histological changes which were partially affected by the drug. Clonidine reduced the increase in BUN and serum creatinine concentration in envenomed mice. Td venom produced neutrophilia and lymphopenia, which were clonidine-resistant at the assayed dose. These results suggest that α(2)-adrenoceptor selective agonists would be able to reduce some scorpion venom-induced renal and pancreatic disturbances, possibly through the inhibition of neurotransmitter release from presynaptic cholinergic and noradrenergic terminals, as well as from adrenal medulla.


Assuntos
Agonistas de Receptores Adrenérgicos alfa 2/farmacologia , Clonidina/farmacologia , Venenos de Escorpião/toxicidade , Escorpiões/fisiologia , Animais , Nitrogênio da Ureia Sanguínea , Creatinina/sangue , Sinergismo Farmacológico , Precursores Enzimáticos , Hiperglicemia/sangue , Hiperglicemia/induzido quimicamente , Processamento de Imagem Assistida por Computador , Rim/efeitos dos fármacos , Rim/patologia , Contagem de Leucócitos , Leucócitos/efeitos dos fármacos , Leucócitos/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pâncreas/efeitos dos fármacos , Pâncreas/metabolismo , Pancreatite/induzido quimicamente , Pancreatite/patologia , Vesículas Secretórias/efeitos dos fármacos , Vesículas Secretórias/ultraestrutura , alfa-Amilases/sangue , alfa-Amilases/urina
16.
J Immunotoxicol ; 9(2): 173-83, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22424317

RESUMO

In Brazil, the species Tityus serrulatus is responsible for the most severe cases of scorpion envenomation. There is currently a need for new scorpion anti-venoms that are more effective and less harmful. This study attempted to produce human monoclonal antibodies capable of inhibiting the activity of T. serrulatus venom (TsV), using the Griffin.1 library of human single-chain fragment-variable (scFv) phage antibodies. Four rounds of phage antibody selection were performed, and the round with the highest phage antibody titer was chosen for the production of monoclonal phage antibodies and for further analysis. The scFv 2A, designated serrumab, was selected for the production and purification of soluble antibody fragments. In a murine peritoneal macrophage cell line (J774.1), in vitro assays of the cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-10 were performed. In male BALB/c mice, in vivo assays of plasma urea, creatinine, aspartate transaminase, and glucose were performed, as well as of neutrophil recruitment and leukocyte counts. It was found that serrumab inhibited the TsV-induced increases in the production of IL-6, TNFα, and IL-10 in J774.1 cells. The in vivo inhibition assay showed that serrumab also prevented TsV-induced increases in the plasma levels of urea, creatinine, aspartate transaminase, and glucose, as well as preventing the TsV-induced increase in neutrophil recruitment. The results indicate that the human monoclonal antibody serrumab is a candidate for inclusion in a mixture of specific antibodies to the various toxins present in TsV. Therefore, serrumab shows promise for use in the production of new anti-venom.


Assuntos
Anticorpos Monoclonais Humanizados/imunologia , Antivenenos/imunologia , Proteínas de Insetos/imunologia , Venenos de Escorpião/imunologia , Escorpiões/fisiologia , Anticorpos de Cadeia Única/imunologia , Animais , Anticorpos Monoclonais Humanizados/biossíntese , Anticorpos Monoclonais Humanizados/uso terapêutico , Antivenenos/biossíntese , Antivenenos/uso terapêutico , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Proteínas de Insetos/antagonistas & inibidores , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Testes de Neutralização , Biblioteca de Peptídeos , Venenos de Escorpião/antagonistas & inibidores , Venenos de Escorpião/toxicidade , Anticorpos de Cadeia Única/biossíntese , Anticorpos de Cadeia Única/uso terapêutico
17.
J Hered ; 103(2): 221-9, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22312118

RESUMO

When low dispersal ability of an organism meets geographical barriers, the evolution of inter- and intraspecific differentiation is often facilitated. In the Atlas massif of North Africa, the genus Buthus splits into several species and diverges into numerous genetic lineages, often following the orographic structures of mountain systems. Such high mountain ranges often act as barriers for species with reduced mobility even on small spatial scales. To study the effect of orographic structures on organisms with low dispersal ability, we collected 61 individuals of the scorpion species Buthus elmoutaouakili at 18 locations around the southwestern foothills of the High Atlas and Antiatlas and in the Sousse valley (western Morocco). We analyzed intraspecific differentiation patterns within this geographically restricted area of about 100 × 50 km using 452 bp of the cytochrome oxidase I mitochondrial gene. We detected 5 distinct genetic lineages. In a second analysis, we added 61 previously published sequences from Buthus species from Europe and North Africa. Using a molecular clock approach, we detected old splits (4-5 Ma) separating the samples from 1) the western High Atlas and north of these mountains, 2) the Sousse valley and adjoining mountain areas, and 3) the southwestern Antiatlas. Further differentiation happened in the first 2 geographical groups about 3 Ma. Thus, the divergence time estimates based on a Bayesian approach support the onset of differentiation into these main clades along the Pliocene (5-2.3 Ma) when climatic oscillations started and a constant global cooling preceded the glacial-interglacial cycles of the Pleistocene. Further genetic splits into parapatric groups are detectable for the Sousse valley main group in the early Pleistocene. The climatic oscillations of the Pliocene and early Pleistocene might have caused repeated range shifts, expansions, and retractions leading to repeated vicariance, hereby producing the hierarchical structure of genetic differentiation in B. elmoutaouakili. A taxonomic revision, including morphological and molecular data, is needed to assess the status of each of these Buthus scorpion lineages.


Assuntos
Evolução Biológica , Demografia , Evolução Molecular , Variação Genética , Filogenia , Escorpiões/genética , Animais , Sequência de Bases , Teorema de Bayes , Primers do DNA/genética , Haplótipos/genética , Funções Verossimilhança , Modelos Genéticos , Dados de Sequência Molecular , Marrocos , Filogeografia , Escorpiões/fisiologia , Análise de Sequência de DNA
18.
BMC Genomics ; 11: 452, 2010 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-20663230

RESUMO

BACKGROUND: Lychas mucronatus is one scorpion species widely distributed in Southeast Asia and southern China. Anything is hardly known about its venom components, despite the fact that it can often cause human accidents. In this work, we performed a venomous gland transcriptome analysis by constructing and screening the venom gland cDNA library of the scorpion Lychas mucronatus from Yunnan province and compared it with the previous results of Hainan-sourced Lychas mucronatus. RESULTS: A total of sixteen known types of venom peptides and proteins are obtained from the venom gland cDNA library of Yunnan-sourced Lychas mucronatus, which greatly increase the number of currently reported scorpion venom peptides. Interestingly, we also identified nineteen atypical types of venom molecules seldom reported in scorpion species. Surprisingly, the comparative transcriptome analysis of Yunnan-sourced Lychas mucronatus and Hainan-sourced Lychas mucronatus indicated that enormous diversity and vastly abundant difference could be found in venom peptides and proteins between populations of the scorpion Lychas mucronatus from different geographical regions. CONCLUSIONS: This work characterizes a large number of venom molecules never identified in scorpion species. This result provides a comparative analysis of venom transcriptomes of the scorpion Lychas mucronatus from different geographical regions, which thoroughly reveals the fact that the venom peptides and proteins of the same scorpion species from different geographical regions are highly diversified and scorpion evolves to adapt a new environment by altering the primary structure and abundance of venom peptides and proteins.


Assuntos
Perfilação da Expressão Gênica , Variação Genética , Venenos de Escorpião/genética , Escorpiões/genética , Adaptação Fisiológica , Sequência de Aminoácidos , Animais , Evolução Molecular , Etiquetas de Sequências Expressas/metabolismo , Feminino , Masculino , Dados de Sequência Molecular , Família Multigênica , Neurotoxinas/química , Neurotoxinas/genética , Peptídeos/química , Peptídeos/genética , Venenos de Escorpião/química , Escorpiões/fisiologia , Análise de Sequência de DNA , Especificidade da Espécie
19.
Av. cardiol ; 29(1): 68-75, mar. 2009. graf
Artigo em Espanhol | LILACS | ID: lil-607920

RESUMO

Se presentan características clínicas, epidemiológicas y taxonómicas del emponzoñamiento escorpiónico en Venezuela. Se revisan aspectos de la fisiopatología de la venina en el área cardiovascular y las bases racionales del tratamiento. Finalmente, se presentan las pautas del protocolo del Centro Toxicológico Regional "Dra. EL Bermúdez" para la terapia del emponzoñamiento escorpiónico en el Hospital pediátrico "Dr. Agustin Zubillaga".


Clinical, epidemiological and taxonomic features about scorpion envenoming in Venezuela are presented. Aspects about physiopatology of the venom in the cardiovascular area and the rationale of the treatment are reviewed. Finally, the guidelines of "Dra. EL bermúdez" Regional Poison Center for therapy of the scorpion envenoming in "Dr. Agustin Zubillaga" Pediatric Hospital are presented.


Assuntos
Humanos , Masculino , Feminino , Recém-Nascido , Lactente , Criança , Eletrocardiografia/métodos , Doenças Cardiovasculares/etiologia , Escorpiões/classificação , Escorpiões/fisiologia , Exames Médicos/métodos , Peçonhas/intoxicação , Pressão Sanguínea/fisiologia , Dor no Peito/diagnóstico , Toxicidade/métodos , Venezuela
20.
Toxicon ; 53(1): 129-34, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19027774

RESUMO

Scorpion venoms are rich resources of bioactive peptides with extreme variability. Multiple molecular mechanisms are involved in the diversity of scorpion venom peptides. However, alternative splicing, which plays a major role in the generation of proteomic and functional diversity in metazoan organisms, hasn't been reported in genes coding for scorpion venom peptides. In the EST analysis of venom peptide transcripts from scorpion Lychas mucronatus, we reported an alternative splicing event. Transcripts of LmTxLP11 and LmVP1.1 share identical 5' region. LmVP1.1 is a novel type of scorpion venom peptides constrained by one disulfide bridge, whereas LmTxLP11 is an extended version of LmVP1.1. By transcript alignment with its genomic sequence, it is found that both transcripts are generated from a single gene by alternative poly A site and terminal exon. The gene encoding LmTxLP11 and LmVP1.1 is the first one harboring three introns ever reported from scorpion venoms. This work demonstrates for the first time that alternative splicing is involved in regulating the diversity of scorpion venom peptides.


Assuntos
Processamento Alternativo , Genômica , Peptídeos/química , Peptídeos/genética , Venenos de Escorpião/química , Transcrição Gênica , Sequência de Aminoácidos , Animais , Sequência de Bases , Dados de Sequência Molecular , Peptídeos/metabolismo , Escorpiões/fisiologia , Toxinas Biológicas/química , Toxinas Biológicas/genética , Toxinas Biológicas/metabolismo
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