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Sample records for bothrops alternatus increases

  1. Effect of Bothrops alternatus snake venom on macrophage phagocytosis and superoxide production: participation of protein kinase C

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    SS Setubal

    2011-01-01

    Full Text Available Envenomations caused by different species of Bothrops snakes result in severe local tissue damage, hemorrhage, pain, myonecrosis, and inflammation with a significant leukocyte accumulation at the bite site. However, the activation state of leukocytes is still unclear. According to clinical cases and experimental work, the local effects observed in envenenomation by Bothrops alternatus are mainly the appearance of edema, hemorrhage, and necrosis. In this study we investigated the ability of Bothrops alternatus crude venom to induce macrophage activation. At 6 to 100 ¼g/mL, BaV is not toxic to thioglycollate-elicited macrophages; at 3 and 6 ¼g/mL, it did not interfere in macrophage adhesion or detachment. Moreover, at concentrations of 1.5, 3, and 6 ¼g/mL the venom induced an increase in phagocytosis via complement receptor one hour after incubation. Pharmacological treatment of thioglycollate-elicited macrophages with staurosporine, a protein kinase (PKC inhibitor, abolished phagocytosis, suggesting that PKC may be involved in the increase of serum-opsonized zymosan phagocytosis induced by BaV. Moreover, BaV also induced the production of anion superoxide (O2_ by thioglycollate-elicited macrophages. This BaV stimulated superoxide production was abolished after treating the cells with staurosporine, indicating that PKC is an important signaling pathway for the production of this radical. Based on these results, we suggest that phagocytosis and reactive oxygen species are involved in the pathogenesis of local tissue damage characteristic of Bothrops spp. envenomations.

  2. Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom

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    J. O. Costa

    2007-01-01

    Full Text Available Two fibrinogenolytic enzymes, Bothrops alternatus metalloprotease isoform (BaltMP-I and II, were purified from Bothrops alternatus venom using Diethylaminoethyl (DEAE Sephacel, Sephadex G-75 and Heparin-Agarose column chromatography. Purified BaltMP-I and II ran as single protein bands on analytical polyacrylamide gel electrophoresis and showed molecular weights of 29000 and 36000, respectively, under reducing conditions in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. BaltMP-II, but not BaltMP-I, displayed blood-clotting activity in bovine plasma, which was about 10-fold higher than that of the crude venom. Both enzymes were proteolytically active against bovine fibrinogen as substrate. When fibrinogen and each enzyme were incubated at 37°C, at a ratio of 1:100 (w/w, BaltMP-II cleaved preferentially the Aalpha -chain and more slowly the Bbeta -chain. The action of BaltMP-I was similar, but lower. None of the proteases degraded the gamma-chain of fibrinogen. The fibrinogenolytic activity of the enzymes was inhibited by 1,10-phenanthroline, suggesting they are metalloproteases. Since both enzymes were found to cause defibrinogenation when intraperitoneally (i.p. administered to mice, they can be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.

  3. A transcriptomic analysis of gene expression in the venom gland of the snake Bothrops alternatus (urutu

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    Menossi Marcelo

    2010-10-01

    Full Text Available Abstract Background The genus Bothrops is widespread throughout Central and South America and is the principal cause of snakebite in these regions. Transcriptomic and proteomic studies have examined the venom composition of several species in this genus, but many others remain to be studied. In this work, we used a transcriptomic approach to examine the venom gland genes of Bothrops alternatus, a clinically important species found in southeastern and southern Brazil, Uruguay, northern Argentina and eastern Paraguay. Results A cDNA library of 5,350 expressed sequence tags (ESTs was produced and assembled into 838 contigs and 4512 singletons. BLAST searches of relevant databases showed 30% hits and 70% no-hits, with toxin-related transcripts accounting for 23% and 78% of the total transcripts and hits, respectively. Gene ontology analysis identified non-toxin genes related to general metabolism, transcription and translation, processing and sorting, (polypeptide degradation, structural functions and cell regulation. The major groups of toxin transcripts identified were metalloproteinases (81%, bradykinin-potentiating peptides/C-type natriuretic peptides (8.8%, phospholipases A2 (5.6%, serine proteinases (1.9% and C-type lectins (1.5%. Metalloproteinases were almost exclusively type PIII proteins, with few type PII and no type PI proteins. Phospholipases A2 were essentially acidic; no basic PLA2 were detected. Minor toxin transcripts were related to L-amino acid oxidase, cysteine-rich secretory proteins, dipeptidylpeptidase IV, hyaluronidase, three-finger toxins and ohanin. Two non-toxic proteins, thioredoxin and double-specificity phosphatase Dusp6, showed high sequence identity to similar proteins from other snakes. In addition to the above features, single-nucleotide polymorphisms, microsatellites, transposable elements and inverted repeats that could contribute to toxin diversity were observed. Conclusions Bothrops alternatus venom gland

  4. Kinetics of venom and antivenom serum and clinical parameters and treatment efficacy in Bothrops alternatus envenomed dogs.

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    Jacome, Do; Melo, M M; Santos, M M B; Heneine, L G D

    2002-12-01

    Dogs envenomed with non-lethal doses of Bothrops alternatus venom received standard antivenom therapy, im injections of flunixin meglumine, or topical treatmentwith aqueous Curcuma longa plant extract. Biodistribution of the venom and antivenom were determined by ELISA. There was no significant difference in the efficacy of antivenom and plant extract on local effects; flunixin treatment had lower efficacy. Distribution of the venom was similar with all 3 treatments. Serum levels of the antivenom reached maximum 2-4 h after administration and were not detected after the 5th d.

  5. Treatment of Bothrops alternatus envenomation by Curcuma longa and Calendula officinalis extracts and ar-turmerone Tratamento local do envenenamento por Bothrops alternatus com extrato de Curcuma longa e Calendula officinalis e ar-turmerone

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    M.M. Melo

    2005-02-01

    Full Text Available It was investigated the efficiency of two extracts of plants and one fraction of their properties against the local effects of bothropic envenomation. Bothrops alternatus venom (1.25µg diluted in 100µl of sterile saline solution was inoculated (intradermally into the shaved dorsal back skin of 30 New Zealand rabbits. The animals were divided in six groups receiving the following treatments: group I: subcutaneous application of Curcuma longa extract (1.0ml; group II: topic treatment of Curcuma longa hydroalcoholic extract (1.0ml; group III: topic application of ar-turmerone in vaseline (1.0g; group IV: topic application of Curcuma longa methanolic extract (1.0ml; group V: topic application of Calendula officinalis ointment (1.0g; group VI: topic application of saline (1.0ml. These treatments were done at 30 minutes, and at 2, 4, 24 and 72 hours after venom inoculation. Intensity of local edema, hemorrhagic halo and necrosis were evaluated until 168h after that. Additionally, seven days after the Bothrops venom inoculation, blood was collected from heart with and without EDTA (10% for hemogram and biochemical parameters (total protein, blood urea nitrogen, creatinine, and fibrinogen and all the animals were anesthetized, sacrificed by ether inhalation and submitted to necropsy. Fragments of tissues were taken for histopathological evaluation. The most efficient treatment for inhibition of edema, necrosis and local hemorrhage after Bothrops alternatus venom was the topic application of ar-turmerone.Investigou-se a eficácia do extrato de plantas no tratamento local do envenenamento botrópico. Veneno de serpentes Bothrops alternatus (1,25µg diluído em 100µl de solução salina estéril foi inoculado (via intradérmica entre as escápulas de 30 coelhos. Os animais foram divididos em seis grupos (tratamentos: grupo I: tratamento subcutâneo com extrato de Curcuma longa; grupo II: tratamento tópico com extrato hidroalcoólico de Curcuma longa

  6. Aspectos clínico-patológicos e laboratoriais do envenenamento experimental por Bothrops alternatus em bovinos Clinic and pathological and laboratory aspects of experimental poisoning by Bothrops alternatus venom in cattle

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    Saulo A. Caldas

    2008-06-01

    , acompanhada de hemorragia, no entorno do local da inoculação nos animais que receberam o veneno por via intramuscular; essa lesão era discreta nos músculos próximos ao local de inoculação subcutânea. Nos bovinos deste estudo, o aumento de volume observado no local de inoculação e adjacências era constituído por sangue e não edema. Não foram observadas mioglobinúria, nem lesões macro ou microscópicas significativas nos rins. Este estudo indica que exemplares de B. alternatus, caso inoculem todo seu veneno, poderiam levar bovinos adultos à morte. Por outro lado, pelo fato de ofídios serem capazes de regular a quantidade de veneno inoculada e, possivelmente, não considerarem bovinos como presa potencial, é provável que o número de acidentes nessa espécie seja pequeno, o que está de acordo com o observado na maioria dos centros diagnóstico anátomo-patológico no país.The aim of this study was to determine the clinical-pathological alterations and laboratory findings in cattle inoculated with Bothrops alternatus venom, with the intention of providing information for the establishment of diagnosis and differential diagnosis procedures, as well as to elucidate some obscurities observed in the pertinent literature. The lyophilized venom was diluted in 1 ml of physiologic solution. It was administered to 5 bovines by the subcutaneous route at doses of 0.0625, 0.125 and 0.25mg/kg body weight, and to 2 bovines by the intramuscular route at doses of 0.25 e 0.45mg/kg. Six bovines died and the only animal that survived, who had subcutaneously received the venom at a dose of 0.0625mg/kg, recovered. The first clinical signs were observed from 25min to 5h30min after the inoculation. The clinical evolution time varied from 7 hours 18 minutes to 92 hours. Regardless of the dose, the clinical picture was characterized by swelling (hemorrhage/hematoma at the site of inoculation, increase in bleeding time and capillary refill time, paleness of mucous membranes and

  7. Heterologous expression and biochemical and functional characterization of a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus snake.

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    Santos-Filho, Norival A; Boldrini-França, Johara; Santos-Silva, Ludier K; Menaldo, Danilo L; Henrique-Silva, Flávio; Sousa, Tiago S; Cintra, Adélia C O; Mamede, Carla C N; Oliveira, Fábio; Arantes, Eliane C; Antunes, Lusânia M Greggi; Cilli, Eduardo M; Sampaio, Suely V

    2014-10-01

    Venomous and non-venomous snakes possess phospholipase A2 (PLA2) inhibitory proteins (PLIs) in their blood serum. This study shows the expression and biochemical and functional characterization of a recombinant alpha inhibitor from Bothrops alternatus snake, named rBaltMIP. Its expression was performed in Pichia pastoris heterologous system, resulting in an active recombinant protein. The expressed inhibitor was tested regarding its ability to inhibit the phospholipase activity of different PLA2s, showing slight inhibitions especially at the molar ratios of 1:1 and 1:3 (PLA2:PLI). rBaltMIP was also effective in decreasing the myotoxic activity of the tested toxins at molar ratios greater than 1:0.4 (myotoxin:PLI). The inhibition of the myotoxic activity of different Asp49 (BthTX-II and PrTX-III) and Lys49 (BthTX-I and PrTX-I) myotoxins was also performed without the prior incubation of myotoxins/inhibitor in order to analyze the real possibility of using snake plasma inhibitors or recombinant inhibitors as therapeutic agents for treating envenomations. As a result, rBaltMIP was able to significantly inhibit the myotoxicity of Lys49 myotoxins. Histopathological analysis of the gastrocnemius muscles of mice showed that the myotoxins are able to induce severe damage to the muscle fibers of experimental animals by recruiting a large number of leukocyte infiltrates, besides forming an intense accumulation of intercellular fluid, leading to local edema. When those myotoxins were incubated with rBaltMIP, a reduction of the damage site could be observed. Furthermore, the cytotoxic activity of Asp49 PLA2s and Lys49 PLA2-like enzymes on C2C12 cell lines was decreased, as shown by the higher cell viabilities after preincubation with rBaltMIP. Heterologous expression would enable large-scale obtainment of rBaltMIP, thus allowing further investigations for the elucidation of possible mechanisms of inhibition of snake PLA2s, which have not yet been fully clarified. PMID:25047442

  8. Caracterização individual do veneno de Bothrops alternatus Duméril, Bibron & Duméril em função da distribuição geográfica no Brasil (Serpentes,Viperidae Individual characterization of Bothrops alternatus Duméril, Bibron & Duméril venoms, according to their geographic distribution in Brazil (Serpentes, Viperidae

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    Marisa M. T. da Rocha

    2005-06-01

    Full Text Available Bothrops alternatus Duméril, Bibron & Duméril, 1854 é uma serpente de importância em saúde pública, com ampla distribuição geográfica, desde o Mato Grosso do Sul até o sudeste do Brasil, chegando até a Argentina e Uruguai, ocupando vários domínios morfoclimáticos. Neste trabalho investigou-se a variação do veneno de adultos de Bothrops alternatus, em função de sua distribuição geográfica no Brasil, comparativamente ao veneno elaborado sob a forma de "pool" desta espécie (veneno referência, que inclui serpentes, em sua maioria, da região do estado de São Paulo. Foram analisadas as atividades letal, coagulante sobre o plasma, proteolítica sobre a caseína e miotóxica, bem como os padrões eletroforéticos de 61 amostras individuais de veneno contrapostas ao "pool". Os resultados mostraram que o veneno de B. alternatus é pouco ativo, comparativamente ao de outros Bothrops Wagler, 1824. A variação individual prevaleceu, não apresentando correlação com as áreas de distribuição geográfica e domínios morfoclimáticos, porém a atividade coagulante das amostras de veneno provenientes do nordeste da distribuição geográfica apresentaram-se menos ativas comparativamente às da porção central da distribuição. Os venenos provenientes das bordas da distribuição apresentaram ações proteolíticas e miotóxicas mais intensas, que estatisticamente não foram significativamente diferentes. As variações individuais prevaleceram.Bothrops alternatus Duméril, Bibron & Duméril, 1854 snakebites are an important public health problem in Brazil. Such snakes are found from Mato Grosso do Sul (central Brazil to southeastern Brazil, reaching even Argentina and Uruguay and thereby occupying different morphoclimatic domains. This work investigated venom variation occurring in adult specimens of B. alternatus specimens, according to their geographic distribution in Brazil. The standard venom pool (reference venom produced by

  9. Comparación de caracteres corporales y del veneno de Bothrops alternatus entre poblaciones de las provincias de Buenos Aires y Entre Ríos, Argentina

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    Rocco, Daniela Marisa

    2011-11-01

    Full Text Available Comparamos caracteres corporales y producción de veneno de ejemplares de Bothrops alternatus de una población aislada geográficamente (Olavarría, región de Tandilia, Buenos Aires con otra en su área de distribución continua de Concordia (Entre Ríos. Estudiamos el largo corporal, peso, separación entre dientes inoculadores, cantidad de veneno y de proteínas en el veneno por ejemplar. No se hallaron diferencias en los caracteres estudiados entre ambas poblaciones (p > 0.05. Las hembras fueron mayores que los machos en ambas muestras, entre un 12-18% (p 0.5; Olavarría: 142 ± 65 mg/animal, Concordia: 160 ± 80 mg/animal, aún ajustando la cantidad de veneno producida respecto al tamaño, mediante el cociente veneno/largo corporal (p >0.6. Tampoco hubo diferencias en el contenido proteico, siendo para ambas muestras de 0.697 ± 0.096 mg de proteínas/mg de veneno seco. Nuestros datos sugieren que los ejemplares de la población aislada de Tandilia no presentan variaciones en el tamaño corporal o en la cantidad de veneno producida, respecto a los ejemplares de Concordia. Some corporal characters and venom yield of adult specimens of Bothrops alternatus from the isolated region of Olavarría (Tandilian Region, Buenos Aires and the region of continuous distribution of Concordia (Entre Ríos were compared. Corporal length, weight, separation between fangs, venom yield by snake and the amount of protein in the venom were determined. No differences in the distinct characters from specimens from both regions were found (p > 0.05. Females were greater than males in both samples, 12-18% (p 0.5; Olavarría= 142 ± 65 mg / animal, Concordia= 161 ± 80 mg / animal even adjusting the venom yield with the corporal length by the relation mg of venom / corporal length (p > 0.6. No differences were observed in the protein content of the dry venom which was of 0.697 ± 0.096 mg of protein / mg of dry venom. From the study of these samples, it could be

  10. Avaliação das lesões locais de cães envenenados experimentalmente com Bothrops alternatus após diferentes tratamentos Evaluation of local lesions in dogs experimentally envenomed by Bothrops alternatus after different treatments

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    M.M.B. Santos

    2003-10-01

    Full Text Available The aim of this study was to evaluate the therapeutic action of flunixin meglumine and aqueous extract of Curcuma longa (10% on local lesions caused by bothropic venom, comparing with the specific antivenom. Twelve adult dogs were inoculated in the middle third of the lateral face of the thigh with 0.3mg/kg of venom. The dogs were divided into three groups of four animals and the treatment was done two hours after venom inoculation as follows: Group I- specific antivenom diluted in saline (one dosage sufficient to neutralize the venom, Group II- flunixin meglumine (1.1mg/kg, IM, once a day for five days, Group III- topical application of aqueous extract of C. longa (10% (three times a day for five days. Necrosis was observed in all dogs of groups I and II, and in two dogs of group III. Edema and hemorrhagic halo were observed in all dogs. Better recovery of lesions was observed in dogs of group I and III. The results denote a satisfactory effect of C. longa on treatment of local lesions caused by bothropic venom in dogs.

  11. Alternagin-C, a disintegrin-like protein from the venom of Bothrops alternatus, modulates a2ß1 integrin-mediated cell adhesion, migration and proliferation

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    Selistre-de-Araujo H.S.

    2005-01-01

    Full Text Available The alpha2ß1 integrin is a major collagen receptor that plays an essential role in the adhesion of normal and tumor cells to the extracellular matrix. Alternagin-C (ALT-C, a disintegrin-like protein purified from the venom of the Brazilian snake Bothrops alternatus, competitively interacts with the alpha2ß1 integrin, thereby inhibiting collagen binding. When immobilized in plate wells, ALT-C supports the adhesion of fibroblasts as well as of human vein endothelial cells (HUVEC and does not detach cells previously bound to collagen I. ALT-C is a strong inducer of HUVEC proliferation in vitro. Gene expression analysis was done using an Affimetrix HU-95A probe array with probe sets of ~10,000 human genes. In human fibroblasts growing on collagen-coated plates, ALT-C up-regulates the expression of several growth factors including vascular endothelial growth factor, as well as some cell cycle control genes. Up-regulation of the vascular endothelial growth factor gene and other growth factors could explain the positive effect on HUVEC proliferation. ALT-C also strongly activates protein kinase B phosphorylation, a signaling event involved in endothelial cell survival and angiogenesis. In human neutrophils, ALT-C has a potent chemotactic effect modulated by the intracellular signaling cascade characteristic of integrin-activated pathways. Thus, ALT-C acts as a survival factor, promoting adhesion, migration and endothelial cell proliferation after binding to alpha2ß1 integrin on the cell surface. The biological activities of ALT-C may be helpful as a therapeutic strategy in tissue regeneration as well as in the design of new therapeutic agents targeting alpha2ß1 integrin.

  12. Estudo histopatológico das lesões causadas pelo veneno de urutu (Bothrops alternatus em músculo esquelético de camundongos

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    L. S. Queiroz

    1984-10-01

    Full Text Available Veneno bruto de urutu (Bothrops alternatus dissolvido em solução salina fisiológica foi injetado no músculo tibial anterior direito de camundongos adultos na dose de 80 μg. Os músculos foram examinados em cortes de parafina, corados por Hematoxilina e Eosina. Aos 10 minutos já havia intensa hemorragia difusa no M. tibial anterior, mas apenas raras fibras musculares estavam necróticas. Nas horas seguintes, contudo, observou-se rápido aumento do número de fibras afetadas, sendo que às 24 hs o músculo apresentava-se totalmente necrótico. Vasos sangüíneos intramusculares e nas proximidades do M. tibial anterior mostravam necrose hialina da camada média e por vezes trombose. A fagocitose dos restos celulares ocorreu da periferia para o centro e acompanhou-se de regeneração muscular. Após 1 a 2 meses, em vários animais houve recuperação considerável do músculo, embora com persistência de cicatriz. As fibras regeneradas possuiam núcleos centrais e variavam em diâmetro, estando muitas atróficas. Em outros camundongos a regeneração do M. tibial anterior foi muito precária, tendo este sido substituído por tecido fibroadiposo com apenas raras fibras musculares. Os resultados mostram que, apesar da gravidade das lesões iniciais devidas ao veneno, ocorre regeneração muscular em grau variável de animal para animal. Sugere-se que a má regeneração observada em alguns casos poderia ser devida, ao menos em parte, a dano vascular permanente.

  13. The defensive strike of five species of lanceheads of the genus Bothrops (Viperidae).

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    Araújo, M S; Martins, M

    2007-05-01

    We studied the defensive strike of one species of each of five recognized lineages within the genus Bothrops, namely, B. alternatus, B. jararaca, B. jararacussu, B. moojeni and B. pauloensis. The defensive strike of the studied species was in general similar to that of Crotalus viridis and C. atrox, but some important differences were observed. Bothrops alternatus and B. pauloensis struck preferentially from a tight body posture, whereas B. jararaca and B. moojeni from a loose body posture. Defensive strikes were either true or false (during the latter, the mouth remains closed or partially open). Almost all strikes were successful; only on a few occasions snakes missed their target (flawed strikes). Strike variables were very conservative among the five species, especially strike distance and height, and one possible explanation may be related to constraints imposed on strike variables as a way of increasing strike accuracy. PMID:17876444

  14. Bothrops jararaca venom metalloproteinases are essential for coagulopathy and increase plasma tissue factor levels during envenomation.

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    Karine M Yamashita

    2014-05-01

    Full Text Available BACKGROUND/AIMS: Bleeding tendency, coagulopathy and platelet disorders are recurrent manifestations in snakebites occurring worldwide. We reasoned that by damaging tissues and/or activating cells at the site of the bite and systemically, snake venom toxins might release or decrypt tissue factor (TF, resulting in activation of blood coagulation and aggravation of the bleeding tendency. Thus, we addressed (a whether TF and protein disulfide isomerase (PDI, an oxireductase involved in TF encryption/decryption, were altered in experimental snake envenomation; (b the involvement and significance of snake venom metalloproteinases (SVMP and serine proteinases (SVSP to hemostatic disturbances. METHODS/PRINCIPAL FINDINGS: Crude Bothrops jararaca venom (BjV was preincubated with Na2-EDTA or AEBSF, which are inhibitors of SVMP and SVSP, respectively, and injected subcutaneously or intravenously into rats to analyze the contribution of local lesion to the development of hemostatic disturbances. Samples of blood, lung and skin were collected and analyzed at 3 and 6 h. Platelet counts were markedly diminished in rats, and neither Na2-EDTA nor AEBSF could effectively abrogate this fall. However, Na2-EDTA markedly reduced plasma fibrinogen consumption and hemorrhage at the site of BjV inoculation. Na2-EDTA also abolished the marked elevation in TF levels in plasma at 3 and 6 h, by both administration routes. Moreover, increased TF activity was also noticed in lung and skin tissue samples at 6 h. However, factor VII levels did not decrease over time. PDI expression in skin was normal at 3 h, and downregulated at 6 h in all groups treated with BjV. CONCLUSIONS: SVMP induce coagulopathy, hemorrhage and increased TF levels in plasma, but neither SVMP nor SVSP are directly involved in thrombocytopenia. High levels of TF in plasma and TF decryption occur during snake envenomation, like true disseminated intravascular coagulation syndrome, and might be implicated in

  15. Genotoxic effect of Bothrops snake venoms and isolated toxins on human lymphocyte DNA.

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    Marcussi, Silvana; Stábeli, Rodrigo G; Santos-Filho, Norival A; Menaldo, Danilo L; Silva Pereira, Luciana L; Zuliani, Juliana P; Calderon, Leonardo A; da Silva, Saulo L; Antunes, Lusânia M Greggi; Soares, Andreimar M

    2013-04-01

    In the present study, micronucleus with cytokinesis blocking and comet assays were used to evaluate the genotoxic potential of Bothrops jararacussu, Bothrops atrox, Bothrops moojeni, Bothrops alternatus (Rhinocerophis alternatus) and Bothrops brazili snake venoms, and also of some isolated toxins (MjTX-I, BthTX-I and II myotoxins, BjussuMP-II metalloprotease, and BatxLAAO l-amino acid oxidase) on human lymphocytes. Significant DNA damages were observed, indicating genotoxic potential after exposure of the lymphocytes to the toxins BthTX-I, II and BatxLAAO compared to untreated and Cisplatin-treated controls, which were able to induce greater formation of micronuclei. B. brazili, B. jararacussu and B. atrox crude venoms also presented genotoxic potential, and the latter two induced DNA breakage 5 times more often than in normal environmental conditions (control without treatment). B. jararacussu venom and its isolated toxins, as well as an LAAO from B. atrox, were able to cause lymphocyte DNA breakage in the comet test with more than 85% damage levels. The DNA damage evaluation allows a widening of the toxic-pharmacological characterization of snake venoms and their toxins and also contributes to the understanding of the mechanisms of action of these molecules in several human pathologies. PMID:23333649

  16. Cross-neutralization of the coagulant activity of Crotalus durissus terrificus venom from the northeast of Argentina by bivalent bothropic antivenom

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    JP Rodríguez

    2012-01-01

    Full Text Available Cross-neutralization of Crotalus durissus terrificus venom coagulant activity was tested using bivalent horse antivenom against Bothrops alternatus and Bothrops diporus venoms. Our in vitro and in vivo experiments showed that bothropic antivenom neutralizes the thrombin-like activity of crotalic snake venom and this cross-reaction was demonstrated by immunoassays either with whole venom or a purified thrombin-like enzyme. These results suggest common antigenic properties and, consequently, similar molecular structure among venom thrombin-like enzymes. Besides, they provide information that could be further used in the development of new antivenom formulations.

  17. Bothrops lanceolatus (Fer de lance) venom induces oedema formation and increases vascular permeability in the mouse hind paw.

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    de Araújo, A L; de Souza, A O; da Cruz-Höfling, M A; Flores, C A; Bon, C

    2000-02-01

    The ability of snake venoms to increase vascular permeability and to induce oedema through the release of pharmacologically active substances is well known. We have studied the oedema and vascular permeability induced by Bothrops lanceolatus venom in male Swiss white mice. Paw oedema was induced by the subplantar injection of B. lanceolatus venom (125-1000 ng/paw) and was quantified as the increase in paw weight. Changes in vascular permeability were assessed by measuring the amount of Evans blue dye extravasation. The oedema and the increase in vascular permeability were maximal within 2 h and had resolved after 24 h. The administration of the vasodilator iloprost (20 ng/paw) immediately after B. lanceolatus venom potentiated the oedema and the increase in vascular permeability by approximately four-fold. Pretreating the mice with indomethacin, dexamethasone, NDGA or BW A4C inhibited the venom-induced oedema and the increase in vascular permeability. In contrast, histamine, serotonin and PAF-acether antagonists (mepyramine, cyproheptadine and WEB 2086, respectively) were ineffective. Histological examination showed that B. lanceolatus venom (250 ng and 500 ng/paw) caused thickening of the inner dermal layers which was accompanied by extensive intercellular spaces indicative of oedema. In addition, there was a marked infiltration of inflammatory cells, particularly neutrophils, into the underlying muscle layer. The latter, however, remained morphologically unaffected during the 3 h of observation. Venom doses larger than 500 ng/paw produced intense haemorrhage. These results indicate that B. lanceolatus venom induces oedema and increases vascular permeability in the mouse hind paw. The principal mediators of this inflammatory response are cyclooxygenase and lipoxygenase products. PMID:10665802

  18. Cold tolerance and cold hardening strategy of the Japanese pine sawyer Monochamus alternatus (Coleoptera: Cerambycidae)

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    Jing Tian; Shu-Guang Hao; Wei-Na Kong; Rui-Yan Ma; Le Kang

    2008-01-01

    The Japanese pine sawyer, Monochamus alternatus, is an important pine forest pest and vector transmitting the pine wilt nematode that causes pine wilt disease. Low temperatures in autumn, winter and spring often differentially affect mortality of M.alternatus larvae. In this paper, we mainly compared the differences of mortality and cold hardening of larvae from different seasons, based on supercooling point (SCP) and cumulative probability of individuals freezing (CPIF). The cold hardening of the larvae from autumn, winter and spring seasons were largely different. Correlations between mortality and CPIF of autumn and spring larvae were highest on day 1/4, and gradually decreased with prolonged exposure duration. This beetle's death mainly resulted from freezing in short exposure duration. However, the correlation between mortality and CPIF of winter larvae increased gradually with the prolonged exposure duration. Death did not mainly result from freezing in long exposure duration. Autumn larvae are more susceptible and adaptable than winter and spring larvae. Winter larvae have a slight freeze-tolerance trend. Our research showed that M. alternatus came into complex cold-hardening strategies under natural selection. Freeze avoidance is the primary strategy; with prolonged exposure duration to above SCP or < 0℃, chill tolerance is more important; this is followed by freeze tolerance during harsh winters.

  19. Genetic relationships among species of the genus Bothrops based on RAPD markers

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    Felipe Grazziotin

    2005-05-01

    Full Text Available The genetic relationships among 11 species of Bothrops found in Brazil were determined using random amplified fragment (RAPD information. A total of 239 amplified bands were scored using 20 aleatory primers. Most of the bands (88.7% were polymorphic. Phylogenetic analysis of the data determined three similarity groups within Bothrops: Group I- B. alternatus, B. neuwiedi, B. cotiara, and B. jararacusu; Group II- B. insularis, B. jararaca, and B. erythromelas; and Group III- B. moojeni, B. leucurus, and B. atrox. These groups coincided with those obtained by other methods, indicating that RAPD's could be a useful tool for the evaluation of genetic relationships at the interspecific level.As relações genéticas entre 11 espécies de Bothrops (sensus stricto de ocorrência no Brasil foram determinadas através da comparação dos perfis de fragmentos amplificados aleatoriamente (RAPD. Um total de 239 bandas foram obtidas utilizando 20 sequencias iniciadoras (primers. A maior parte das bandas (88,7% foram polimórficas. A análise filogenética dos dados permitiu a identificação de três grupos dentro do gênero Bothrops: Grupo 1- B. alternatus, B. neuwiedi, B. cotiara, e B. jararacusu; Grupo II- B. insularis, B. jararaca, e B. erythromelas; e Grupo III- B. moojeni, B. leucurus, e B. atrox. Estes grupos coincidem com aqueles obtidos por outros métodos indicando que os marcadores de RAPD podem ser considerados ferramentas úteis para a avaliação de relações a nível interespecífico.

  20. Neuromuscular activity of Bothrops alcatraz snake venom in chick biventer cervicis preparations.

    Science.gov (United States)

    de Moraes, Delkia Seabra; Aparecido de Abreu, Valdemir; Rostelato-Ferreira, Sandro; Leite, Gildo B; Alice da Cruz-Höfling, Maria; Travaglia-Cardoso, Silvia R; Hyslop, Stephen; Rodrigues-Simioni, Léa

    2012-02-01

    Venom (10-100 μg/ml) from Bothrops alcatraz, a pitviper from the Alcatrazes Archipelago off the coast of southeastern Brazil, caused progressive, irreversible neuromuscular blockade in chick isolated biventer cervicis preparations. The venom also inhibited contractures to exogenous ACh (110 μM) and KCl (20 mM), caused myofiber damage and increased creatine kinase release. Commercial bothropic antivenom raised against mainland Bothrops species neutralized the neuromuscular activity, depending on the venom concentration. PMID:22155137

  1. Effects of methamidophos and deltamethrin on in vitro protein phosphorylation in Monochamus alternatus

    Institute of Scientific and Technical Information of China (English)

    Jie Liu; Xi-Wu Gao; Yi-Jun Wu; Wei Li; Qi-Lian Qin; Jiang-Hua Sun

    2008-01-01

    Monochamus alternatus Hope(Coleoptera:Cerambycidae)is not onlY a serious Dest insect to pine trees but alSO the main vector of pine wood nemadote Bursaphelenchus xylophilus,which causes pine wilt disease.To explore the insecticidal mechanism of insecticides to M alternatus,we chose methamidophos and deltamethrin as the representa-tives of two groups of insecticides(organophosphates and pyrethroids),which arc widely used for pest controlin China and investigated their effects on phosphorylation of proteins from the insect.Phosphorylation of proteins from the insect fat body and head was determined by fn vitro 32P-labelling.In the fat body,deltamethtin obviously reduced basal phosphorylation levels of proteins at 111,95,77,and 44 kDa,but enhanced the basal phosphorylation level of a protein at 138 kDa.However,in the presence of calmodulin but not cyclic adenosine monophosphate(cAMP),deitamethrin increased phosphorylation of the protein at 111 kDa.In the head,deltamethrin inhibited basal phosphorylation levels of proteins at 113,98,and 51 kDa,but potentiated phosphorylation of a protein at 167 kDa activated by cAMP.Methamidophos inhibited phosphorylation of a protein at 44 kDa in the fat body.Although methamidophos did not impact basal phosphorylation levels of any proteins in the head,it inhibiled calcium/calmodulin(Ca2+/CAM) stimulated phosphoryla-tion of a protein at 5 1 kDa.Together.our dam indicate that methamidophos and deltamethrin altered phosphorylation levels of various proteins in thc head and fat body of the pine insect and these two kinds of inseeticides acted on the proteins that call be phosphorylated in the tissues respectively,Which is possibly related to their toxicity.

  2. Identification of differentially expressed genes in Monochamus alternatus digested with azadirachtin.

    Science.gov (United States)

    Lin, Tong; Liu, Qisi; Chen, Jingxiang

    2016-01-01

    The pine sawyer beetle Monochamus alternatus Hope, a major forest insect pest, is the primary vector of the destructive forest pest pine wood nematode, Bursaphelenchus xylophilus. Azadirachtin, an active compound of neem, is biologically interesting because it represents a group of important, successful botanical pesticides. We provide insight into the molecular effects of azadirachtin on M. alternatus at the transcriptional level to provide clues about possible molecular-level targets and to establish a link between azadirachtin and insect global responses. We found that 920 and 9894 unique genes were significantly up- and down-regulated, respectively. We obtained expression patterns of the differentially expressed genes (DEGs), identifying 4247, 3488 and 7613 sequences that involved cellular components, molecular functions and biological processes, respectively, and showed that the DEGs were distributed among 50 Gene Ontology categories. The Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that the DEGs were enriched in 50 pathways. Detailed gene profile knowledge of the interaction of azadirachtin with M. alternatus should facilitate the development of more effective azadirachtin-based products against M. alternatus and other target Coleoptera. These results further enhance the value of azadirachtin as a potential insecticide of biological origin, as well as for other biological applications. PMID:27629396

  3. [Bites by snakes in the genus Bothrops: a series of 3,139 cases].

    Science.gov (United States)

    Ribeiro, L A; Jorge, M T

    1997-01-01

    Medical records of 3,139 patients bitten by Bothrops snakes and attended at Vital Brazil Hospital (HVB) from 1981 to 1990 were reviewed. They were more frequent in males (75.7%). In 1,412 cases (45.0%) the snake was classified by species, and 1,376 were B. jararaca, 20 B. jararacussu, 11 B. neuwiedi, 2 B. moojeni, 2 B. alternatus e 1 B. pradoi. The most frequent bitten anatomic regions were: foot (47.5%) and hand (21.3%). Tourniquet was used in 38.2% of the cases and its frequency fell down during the study period (p < 0.05). The clinical features at the bite site were: pain (95.6%), swelling (95.4%), ecchymosis (56.1%), blisters (13.8%), necrosis (16.5%), and abscess (11.0%). Systemic manifestations were: bleeding (12.3%), acute renal failure (1.6%), and shock (0.7%). There were blood coagulation disorders in 1,730 (57.9%) of the 2,990 cases. There were 21 amputations (0.7%) and 9 deaths (0.3%). The average serum dose that was used in treatment fell down during the study period (p < 0.001). PMID:9463193

  4. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming.

    Science.gov (United States)

    Boechat, A L; Paiva, C S; França, F O; Dos-Santos, M C

    2001-01-01

    Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was not effective in neutralizing the desfibrinating and edema-forming activities of B. atrox venom and the hemorrhagic and coagulant actions of both venoms. Heparin diminished the effectiveness of the ABS in the neutralization of the hemorrhagic and edema-forming activities of the B. atrox venom. However, heparin in the 6 IU dose was capable of neutralize the edema-forming of the B. erythromelas and increase the effectiveness of the ABS. Heparin also neutralized the phospholipasic A2 activity of B. atrox (14.3%) and B. erythromelas (28.0%) venoms. For B. erythromelas venom, the associated treatment, heparin and ABS, was more effective in the neutralization of its lethal activity. PMID:11246276

  5. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming

    Directory of Open Access Journals (Sweden)

    BOECHAT Antônio Luiz R.

    2001-01-01

    Full Text Available Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS. It was verified that heparin, in doses of 3 and 6 IU, was not effective in neutralizing the desfibrinating and edema-forming activities of B. atrox venom and the hemorrhagic and coagulant actions of both venoms. Heparin diminished the effectiveness of the ABS in the neutralization of the hemorrhagic and edema-forming activities of the B. atrox venom. However, heparin in the 6 IU dose was capable of neutralize the edema-forming of the B. erythromelas and increase the effectiveness of the ABS. Heparin also neutralized the phospholipasic A2 activity of B. atrox (14.3% and B. erythromelas (28.0% venoms. For B. erythromelas venom, the associated treatment, heparin and ABS, was more effective in the neutralization of its lethal activity.

  6. Host Preference by Monochamus alternatus (Hope) during Maturation Feeding on Pine Species and Masson Pine Provenances

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Host preferences pine of the sawyer beetle, Monochamus alternates (Hope), during maturation feeding on 8 conifer trees and 40 masson pine provenances, were investigated using 3 types of laboratory bioassay of consistent feeding preference, feeding area and visitation frequency. M. alternatus adults have the highest frequency of feeding and prefer to feed on the branches of P. massoniana and P. densiflora and had significant host selectivity on 8 conifer trees in the area of Nanjing. The adult feeding vi...

  7. Acidente por serpentes do gênero Bothrops: série de 3.139 casos

    Directory of Open Access Journals (Sweden)

    Lindioneza Adriano Ribeiro

    1997-12-01

    Full Text Available Em avaliação dos prontuários médicos de 3.139 pacientes picados por serpentes do gênero Bothrops atendidos no Hospital Vital Brazil (HVB, de 1981 a 1990, observou-se maior acometimento do sexo masculino (75,7%. Em 1.412 casos (45,0% a serpente foi identificada, sendo 1.376 B. jararaca, 20 B. jararacussu, 11 B. neuwiedi, 2 B. moojeni, 2 B. alternatus e 1 B. pradoi. As regiões anatômicas mais comumente picadas foram: pé (47,5% e mão (21,3%. O torniquete foi realizado em 38,2% dos casos e sua freqüência diminuiu durante esse período (p Medical records of 3,139 patients bitten by Bothrops snakes and attended at Vital Brazil Hospital (HVB from 1981 to 1990 were reviwed. They were more frequent in males (75.7%. In 1,412 cases (45.0% the snake was classified by species, and 1,376 were B. jararaca, 20 B. jararacussu, 11 B. neuwiedi, 2 B. moojeni, 2 B. alternatus e 1 B. pradoi. The most frequent bitten anatomic regions were: foot (47.5% and hand (21.3%. Tourniquet was used in 38.2% of the cases and its frequency fell down during the study period (p < 0.05. The clinical features at the bite site were: pain (95.6%, swelling (95.4%, echimosis (56.1%, blisters (13.8%, necrosis (16.5%, and abscess (11.0%. Systemic manifestations were: bleeding (12.3%, acute renal failure (1.6%, and shock (0.7%. There were blood coagulation disorders in 1,730 (57.9% of the 2,990 cases. There were 21 amputations (0.7% and 9 deaths (0.3%. The average serum dose that was used in treatment fell down during the study period (p < 0.001.

  8. 21 CFR 864.8100 - Bothrops atrox reagent.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Bothrops atrox reagent. 864.8100 Section 864.8100...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8100 Bothrops atrox reagent. (a) Identification. A Bothrops atrox reagent is a device made from snake venom and used to determine blood...

  9. Postprandial thermogenesis in Bothrops moojeni (Serpentes: Viperidae

    Directory of Open Access Journals (Sweden)

    DR Stuginski

    2011-01-01

    Full Text Available Snakes that can ingest prey that are proportionally large have high metabolic rates during digestion. This great increase in metabolic rate (specific dynamic action - SDA may create a significant augment in the animal's body temperature. The present study investigated postprandial thermogenesis in Bothrops moojeni. Briefly, two groups of snakes were fed meals equivalent to 17 ± 3% and 32 ± 5% of their body weight and were observed for 72 hours, in which thermal images of each snake were taken with an infrared camera in a thermostable environment with a constant air temperature of 30°C. The results showed a significant increase in snake surface temperature, with a thermal peak between 33 and 36 hours after feeding. The meal size had a great impact on the intensity and duration of the thermogenic response. Such increase in temperature appears to be connected with the huge increase in metabolic rates during digestion of relatively large prey by snakes that feed infrequently. The ecologic implication of the thermogenic response is still not well understood; however, it is possible that its presence could affect behaviors associated with the snake digestion, such as postprandial thermophily.

  10. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming

    OpenAIRE

    Antônio Luiz R. BOECHAT; Paiva, Cristiano S.; FRANÇA Francisco Oscar; DOS-SANTOS Maria Cristina

    2001-01-01

    Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was no...

  11. Action of anti-bothropic factor isolated from Didelphis marsupialis on renal effects of Bothrops erythromelas venom.

    Science.gov (United States)

    Martins, Alice M C; Sousa, Fabiola C M; Barbosa, Paulo S F; Toyama, Marcos H; Toyama, Daniela O; Aprígio, Cleidiana C; Queiroz, Maria G R; Guarnieri, Mirian C; Havt, Alexandre; de Menezes, Dalgimar B; Fonteles, Manassés C; Monteiro, Helena S A

    2005-11-01

    Acute renal failure is the most common complication in the lethal cases caused by snakebites in Brazil. Among the Brazilian venom snakes, Bothrops erythromelas is responsible for the majority of accidents in Northeastern Brazil. Didelphis marsupialis serum could inhibit myonecrotic, hemorrhagic, edematogenic hyperalgesic and lethal effects of envenomation determined by ophidian bites. In the present study, we evaluated the action of the anti-bothropic factor isolated from D. marsupialis on the renal effects promoted by B. erythromelas venom without systemic interference. Isolated kidneys from Wistar rats were perfused with Krebs-Henseleit solution containing 6% bovine serum albumin. We analyzed renal perfusion pressure (PP), renal vascular resistance (RVR), glomerular filtration rate (GFR), urinary flow (UF), and the percentages of sodium and potassium tubular transport (%TNa+, %TK+). The B. erythromelas venom (10 microg mL(-1)) decreased the PP (ct = 108.71+/-5.09 mmHg; BE = 65.21+/-5.6 mmHg*) and RVR (ct = 5.76+/-0.65 mmHg mL(-1) g(-1) min(-1); BE = 3.10+/-0.45 mmHg mL(-1) g(-1) min(-1)*). On the other hand, the GFR decreased at 60 min (ct60 = 0.76+/-0.07 mL g(-1) min(-1); BE60 = 0.42+/-0.12 mL g(-1) min(-1)*) and increased at 120 min (ct120 = 0.72+/-0.01 mL g(-1) min(-1); BE120 = 1.24+/-0.26 mL g(-1) min(-1)*). The UF increased significantly when compared with the control group (ct = 0.14+/-0.01 mL g(-1) min(-1); BE = 0.47+/-0.08 mL g(-1) min(-1)*). The venom reduced the %TNa(+) (ct90 = 79.18+/-0.88%; BE90 = 58.35+/-4.86%*) and %TK+ (ct90 = 67.20+/-4.04%; BE90 = 57.32+/-5.26%*) The anti-bothropic factor from D. marsupialis (10 microg mL(-1)) incubated with B. erythromelas venom (10 microg mL(-1)) blocked the effects on PP, RVR, %TNa+, and %TK+, but was not able to reverse the effects in UF and GFR promoted by venom alone. However, the highest concentration of D. marsupialis serum (30 microg mL(-1)) reversed all the renal effects induced by the venom. In

  12. Biochemical and immunological alterations of 60 Co irradiated Bothrops jararacussu venom

    International Nuclear Information System (INIS)

    Proteins irradiation leads to structural alterations resulting in activity and function loss. This process has been useful to detoxify animal venoms and toxins, resulting in low toxicity products which increased immunogenicity. The Bothrops jararacussu venom behaves as a weak immunogen and its lethal activity in not neutralized by either autologous, heterologous or bothropic polyvalent antisera. This venom is markedly myotoxic and and the commercial bothropic antiserum does not neutralize this activity, because of this low immunogenicity of the myotoxins. This present work was done in order to evaluate the possibility of irradiating Bothrops jararacussu, intending to increase the immunogenicity of the myotoxic components, leading to productions of myotoxins neutralizing antibodies. Bothrops jararacussu venom samples were irradiated with 500, 1000 and 2000 Gy of 60 Co gamma rays. A 2.3 folds decrease of toxicity was observed for the 1000 Gy irradiated samples while the 2000 Gy irradiated sample was at least 3.7 folds attenuated. On the other hand, the 500 Gy did not promote any detoxification. Electrophoresis and HPLC data indicate that the irradiation lead to the formation of high molecular weight products (aggregates). The proteolytic and phospholipasic activities decreased in a dose dependent manner, the phospholipases being more resistant than the proteases. Both the animals (rabbit) immunized with either native or 2000 Gy irradiated venom produced native venom binding antibodies, a slightly higher titer being obtained in the serum of the rabbit immunized with the irradiated samples. Western blot data indicate that the anti-irradiated venom Ig Gs recognised a greater amount of either autologous or heterologous venom bands, both sera behaving as genus specific. The anti-native serum did not neutralize the myotoxic activity of native venom, while the anti-irradiated one was able to neutralize this activity. (author)

  13. Trichomoniasis in Bothrops jararaca (serpentes, viperidae)

    OpenAIRE

    F.C. Vilela; M.G. da Silva; T. H. Barrella; R. J. DA SILVA

    2003-01-01

    We describe a case of trichomoniasis in a Bothrops jararaca (Serpentes, Viperidae) donated to the Center for the Study of Venoms and Venomous Animals - CEVAP/UNESP. The animal had diarrhea with great quantity of flagellated protozoa in the feces. Microscopic examination of fecal smears stained with Giemsa revealed the presence of trichomonads, morphologically similar to Trichomonas acosta. Trichomonads were not detected in fecal exams after treatment with a single dose of 40 mg/kg metronidazo...

  14. Trichomoniasis in Bothrops jararaca (serpentes, viperidae

    Directory of Open Access Journals (Sweden)

    F. C. Vilela

    2003-01-01

    Full Text Available We describe a case of trichomoniasis in a Bothrops jararaca (Serpentes, Viperidae donated to the Center for the Study of Venoms and Venomous Animals - CEVAP/UNESP. The animal had diarrhea with great quantity of flagellated protozoa in the feces. Microscopic examination of fecal smears stained with Giemsa revealed the presence of trichomonads, morphologically similar to Trichomonas acosta. Trichomonads were not detected in fecal exams after treatment with a single dose of 40 mg/kg metronidazole (Flagyl®.

  15. Dispersal of the Japanese pine sawyer, Monochamus alternatus (Coleoptera: Cerambycidae), in mainland China as inferred from molecular data and associations to indices of human activity.

    Science.gov (United States)

    Hu, Shao-ji; Ning, Tiao; Fu, Da-ying; Haack, Robert A; Zhang, Zhen; Chen, De-dao; Ma, Xue-yu; Ye, Hui

    2013-01-01

    The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is an important forest pest as well as the principal vector of the pinewood nematode (PWN), Bursaphelenchus xylophilus (Steiner et Buhrer), in mainland China. Despite the economic importance of this insect-disease complex, only a few studies are available on the population genetic structure of M. alternatus and the relationship between its historic dispersal pattern and various human activities. The aim of the present study was to further explore aspects of human activity on the population genetic structure of M. alternatus in mainland China. The molecular data based on the combined mitochondrial cox1 and cox2 gene fragments from 140 individuals representing 14 Chinese populations yielded 54 haplotypes. Overall, a historical (natural) expansion that originated from China's eastern coast to the western interior was revealed by the haplotype network, as well as several recent, long-distant population exchanges. Correlation analysis suggested that regional economic status and proximity to marine ports significantly influenced the population genetic structure of M. alternatus as indicated by both the ratio of shared haplotypes and the haplotype diversity, however, the PWN distribution in China was significantly correlated with only the ratio of shared haplotypes. Our results suggested that the modern logistical network (i.e., the transportation system) in China is a key medium by which humans have brought about population exchange of M. alternatus in mainland China, likely through inadvertent movement of infested wood packaging material associated with trade, and that this genetic exchange was primarily from the economically well-developed east coast of China, westward, to the less-developed interior. In addition, this study demonstrated the existence of non-local M. alternatus in new PWN-infested localities in China, but not all sites with non-local M. alternatus were infested with PWN

  16. Dispersal of the Japanese pine sawyer, Monochamus alternatus (Coleoptera: Cerambycidae, in mainland China as inferred from molecular data and associations to indices of human activity.

    Directory of Open Access Journals (Sweden)

    Shao-ji Hu

    Full Text Available The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae, is an important forest pest as well as the principal vector of the pinewood nematode (PWN, Bursaphelenchus xylophilus (Steiner et Buhrer, in mainland China. Despite the economic importance of this insect-disease complex, only a few studies are available on the population genetic structure of M. alternatus and the relationship between its historic dispersal pattern and various human activities. The aim of the present study was to further explore aspects of human activity on the population genetic structure of M. alternatus in mainland China. The molecular data based on the combined mitochondrial cox1 and cox2 gene fragments from 140 individuals representing 14 Chinese populations yielded 54 haplotypes. Overall, a historical (natural expansion that originated from China's eastern coast to the western interior was revealed by the haplotype network, as well as several recent, long-distant population exchanges. Correlation analysis suggested that regional economic status and proximity to marine ports significantly influenced the population genetic structure of M. alternatus as indicated by both the ratio of shared haplotypes and the haplotype diversity, however, the PWN distribution in China was significantly correlated with only the ratio of shared haplotypes. Our results suggested that the modern logistical network (i.e., the transportation system in China is a key medium by which humans have brought about population exchange of M. alternatus in mainland China, likely through inadvertent movement of infested wood packaging material associated with trade, and that this genetic exchange was primarily from the economically well-developed east coast of China, westward, to the less-developed interior. In addition, this study demonstrated the existence of non-local M. alternatus in new PWN-infested localities in China, but not all sites with non-local M. alternatus were

  17. Assessment of efficacy of bothropic antivenom therapy on microcirculatory effects induced by Bothrops jararaca snake venom.

    Science.gov (United States)

    Battellino, Carolina; Piazza, Roxane; da Silva, Ana M M; Cury, Yara; Farsky, Sandra H P

    2003-04-01

    Intravenous administration of antibothropic antivenom (BAv) neutralises the systemic effects, but does not efficiently reverse the local symptoms elicited by the Bothrops jararaca venom (BjV). The mechanisms involved in this poor protection have not been clarified. In this work, intravital microscopy studies were carried out to determine the efficacy of different schedules of BAv treatment on local effects evoked by topical application of BjV in the microcirculatory network of the internal spermatic fascia of Wistar rats. Results demonstrated that BAv administration 15 min before, simultaneously with, or 15 min after BjV application did not totally reverse the local symptoms, represented by disturbances of coagulation, development of haemorrhage lesions, vascular permeability increase and increment on leukocyte-endothelium interactions. This lack of effectiveness neither reflects an inadequate amount of specific antibodies in the antivenom against toxins responsible for local effects nor an insufficient dose of circulating BAv during the assays. Administration of fluorescein isothiocyanate (FITC) labelled-BAv showed the dynamics of distribution of the antivenom in the microcirculatory network. Images obtained from prior and simultaneously treated animals showed that the antivenom remains at luminal side of vessels before venom application, and the latency time to antivenom leakage is coincidental to that for local effects evoked by the venom. In addition, images from posterior treatment demonstrated that the intense alterations in the microcirculatory network impair antivenom distribution at the site of injection. Together, our data show that the lack of effectiveness of antivenom therapy is due to impaired and delayed venom and antivenom interaction at the site of injury.

  18. Hemostatic properties of Venezuelan Bothrops snake venoms with special reference to Bothrops isabelae venom.

    Science.gov (United States)

    Rodríguez-Acosta, Alexis; Sánchez, Elda E; Márquez, Adriana; Carvajal, Zoila; Salazar, Ana M; Girón, María E; Estrella, Amalid; Gil, Amparo; Guerrero, Belsy

    2010-11-01

    In Venezuela, Bothrops snakes are responsible for more than 80% of all recorded snakebites. This study focuses on the biological and hemostatic characteristics of Bothrops isabelae venom along with its comparative characteristics with two other closely related Bothrops venoms, Bothrops atrox and Bothrops colombiensis. Electrophoretic profiles of crude B. isabelae venom showed protein bands between 14 and 100 kDa with the majority in the range of 14-31 kDa. The molecular exclusion chromatographic profile of this venom contains five fractions (F1-F5). Amidolytic activity evaluation evidenced strong thrombin-like followed by kallikrein-like activities in crude venom and in fractions F1 and F2. The fibrinogenolytic activity of B. isabelae venom at a ratio of 100:1 (fibrinogen/venom) induced a degradation of A alpha and B beta chains at 15 min and 2 h, respectively. At a ratio of 100:10, a total degradation of A alpha and B beta chains at 5 min and of gamma chains at 24 h was apparent. This current study evidences one of rarely reported for Bothrops venoms, which resembles the physiologic effect of plasmin. B. isabelae venom as well as F2 and F3 fractions, contain fibrinolytic activity on fibrin plate of 36, 23.5 and 9.45 mm(2)/microg, respectively using 25 microg of protein. Crude venom and F1 fraction showed gelatinolytic activity. Comparative analysis amongst Venezuelan bothropoid venoms, evidenced that the LD(50) of B. isabelae (5.9 mg/kg) was similar to B. atrox-Puerto Ayacucho 1 (6.1 mg/kg) and B. colombiensis-Caucagua (5.8 mg/kg). B. isabelae venom showed minor hemorrhagic activity, whereas B. atrox-Parguasa (Bolivar state) was the most hemorrhagic. In this study, a relative high thrombin-like activity was observed in B. colombiensis venoms (502-568 mUA/min/mg), and a relative high factor Xa-like activity was found in B. atrox venoms (126-294 mUA/min/mg). Fibrinolytic activity evaluated with 10 microg protein, showed that B. isabelae venom contained higher

  19. Hemostatic properties of Venezuelan Bothrops snake venoms with special reference to Bothrops isabelae venom.

    Science.gov (United States)

    Rodríguez-Acosta, Alexis; Sánchez, Elda E; Márquez, Adriana; Carvajal, Zoila; Salazar, Ana M; Girón, María E; Estrella, Amalid; Gil, Amparo; Guerrero, Belsy

    2010-11-01

    In Venezuela, Bothrops snakes are responsible for more than 80% of all recorded snakebites. This study focuses on the biological and hemostatic characteristics of Bothrops isabelae venom along with its comparative characteristics with two other closely related Bothrops venoms, Bothrops atrox and Bothrops colombiensis. Electrophoretic profiles of crude B. isabelae venom showed protein bands between 14 and 100 kDa with the majority in the range of 14-31 kDa. The molecular exclusion chromatographic profile of this venom contains five fractions (F1-F5). Amidolytic activity evaluation evidenced strong thrombin-like followed by kallikrein-like activities in crude venom and in fractions F1 and F2. The fibrinogenolytic activity of B. isabelae venom at a ratio of 100:1 (fibrinogen/venom) induced a degradation of A alpha and B beta chains at 15 min and 2 h, respectively. At a ratio of 100:10, a total degradation of A alpha and B beta chains at 5 min and of gamma chains at 24 h was apparent. This current study evidences one of rarely reported for Bothrops venoms, which resembles the physiologic effect of plasmin. B. isabelae venom as well as F2 and F3 fractions, contain fibrinolytic activity on fibrin plate of 36, 23.5 and 9.45 mm(2)/microg, respectively using 25 microg of protein. Crude venom and F1 fraction showed gelatinolytic activity. Comparative analysis amongst Venezuelan bothropoid venoms, evidenced that the LD(50) of B. isabelae (5.9 mg/kg) was similar to B. atrox-Puerto Ayacucho 1 (6.1 mg/kg) and B. colombiensis-Caucagua (5.8 mg/kg). B. isabelae venom showed minor hemorrhagic activity, whereas B. atrox-Parguasa (Bolivar state) was the most hemorrhagic. In this study, a relative high thrombin-like activity was observed in B. colombiensis venoms (502-568 mUA/min/mg), and a relative high factor Xa-like activity was found in B. atrox venoms (126-294 mUA/min/mg). Fibrinolytic activity evaluated with 10 microg protein, showed that B. isabelae venom contained higher

  20. Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

    Energy Technology Data Exchange (ETDEWEB)

    Lucas de Oliveira, P.C.; Madruga, R.A.; Barbosa, N.P.U. [Uberaba School of Veterinary Medicine (UNIUBE), MG (Brazil)]. E-mail: pedrolucaso@uol.com.br; Sakate, M. [UNESP, Botucatu, SP (Brazil). School of Veterinary Medicine and Animal Husbandry

    2007-07-01

    Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from {sup 60}Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca) venom, natural and irradiated from a {sup 60}Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5 mg/kg of natural venom; and GIIV, inoculated with 0.5 mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase, aspartate amino-transferase and alanine amino-transferase; clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom. (author)

  1. Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

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    P. C. Lucas de Oliveira

    2007-01-01

    Full Text Available Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from 60Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca venom, natural and irradiated from a 60Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5mg/kg of natural venom; and GIIV, inoculated with 0.5mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase (CK, aspartate amino-transferase (AST and alanine amino-transferase (ALT; clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom.

  2. Effects of Schizolobium parahyba extract on experimental Bothrops venom-induced acute kidney injury.

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    Monique Silva Martines

    Full Text Available BACKGROUND: Venom-induced acute kidney injury (AKI is a frequent complication of Bothrops snakebite with relevant morbidity and mortality. The aim of this study was to assess the effects of Schizolobium parahyba (SP extract, a natural medicine with presumed anti-Bothrops venom effects, in an experimental model of Bothrops jararaca venom (BV-induced AKI. METHODOLOGY: Groups of 8 to 10 rats received infusions of 0.9% saline (control, C, SP 2 mg/kg, BV 0.25 mg/kg and BV immediately followed by SP (treatment, T in the doses already described. After the respective infusions, animals were assessed for their glomerular filtration rate (GFR, inulin clearance, renal blood flow (RBF, Doppler, blood pressure (BP, intra-arterial transducer, renal vascular resistance (RVR, urinary osmolality (UO, freezing point, urinary neutrophil gelatinase-associated lipocalin (NGAL, enzyme-linked immunosorbent assay [ELISA], lactate dehydrogenase (LDH, kinetic method, hematocrit (Hct, microhematocrit, fibrinogen (Fi, Klauss modified and blinded renal histology (acute tubular necrosis score. PRINCIPAL FINDINGS: BV caused significant decreases in GFR, RBF, UO, HcT and Fi; significant increases in RVR, NGAL and LDH; and acute tubular necrosis. SP did not prevent these changes; instead, it caused a significant decrease in GFR when used alone. CONCLUSION: SP administered simultaneously with BV, in an approximate 10∶1 concentration, did not prevent BV-induced AKI, hemolysis and fibrinogen consumption. SP used alone caused a decrease in GFR.

  3. Intradermal injection of Bothrops cotiara venom in mice in an experimental wound model

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    JA Lopes

    2012-01-01

    Full Text Available Bothropic envenomation induces hemorrhage, coagulant disturbances and necrosis. Regarding therapies against the local damage caused by the venom, there is little information on tissue changes until the complete healing. In the current study, local damage was evaluated by examination of morphological inflammatory alterations, mast cell count, and analysis of collagen deposition. Bleeding was evident four hours after inoculation. After 24 hours, a large area of injury appeared presenting disorganized tissue, significant hemorrhage and acute inflammation. After three days, the damaged area was extensive, with a large amount of inflammatory cells and the presence of scab. In seven days, healing and reepithelization process started. And, 21 days later, the epithelium showed less infiltration and no skin appendages. The number of mast cells was similar to control after four hours, with a drop of 50% at 24 hours, followed by an increase until the 21st day. No differences of collagen deposition were observed among experimental groups. Taken together, wound healing after intradermal injection of Bothrops cotiara venom in mice follows similar parameters to wounds caused by other bothropic venoms. The present work reveals the importance of experimental wound models to the study of neutralizing agents against venom toxins.

  4. Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

    International Nuclear Information System (INIS)

    Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from 60Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca) venom, natural and irradiated from a 60Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5 mg/kg of natural venom; and GIIV, inoculated with 0.5 mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase, aspartate amino-transferase and alanine amino-transferase; clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom. (author)

  5. Metabolic Profiling of Somatic Tissues from Monochamus alternatus (Coleoptera: Cerambycidae Reveals Effects of Irradiation on Metabolism

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    Liangjian Qu

    2014-06-01

    Full Text Available A high-level of sexual sterility is of importance for the sterile insect technique (SIT. However, the use of high-dose-intensity gamma radiation to induce sterility has negative impacts not only on reproductive cells but also on somatic cells. In this study, we investigated the metabolite differences in somatic tissues between non-irradiated, 20-Gy-irradiated, and 40-Gy-irradiated male Monochamus alternatus, an important vector of the pathogenic nematode, Bursaphelenchus xylophilus, which kills Asian pines. The results showed that metabolite levels changed moderately in the 20-Gy samples but were markedly altered in the 40-Gy samples compared with the non-irradiated samples. Twenty-six and 53 metabolites were disturbed by 20-Gy and 40-Gy radiation, respectively. Thirty-six metabolites were found to be markedly altered in the 40-Gy samples but were not changed significantly in the 20-Gy samples. The comprehensive metabolomic disorders induced by 40-Gy radiation dysregulated six metabolic pathways involved in the life process. The findings presented in this manuscript will contribute to our knowledge of the characteristic metabolic changes associated with gamma-radiation-induced damage to somatic cells and will allow for better exploration of the SIT for the control of this target pest.

  6. Accidente ofídico causado por Bothrops Asper

    OpenAIRE

    Galofre-Ruiz Mario David

    2013-01-01

    Introduction: The snake bite of the genus Bothrops is an important cause of ophidic accident in Colombia, with high morbidity and mortality. Clinical case: A case of bite by the snake Bothrops Asper, which was classified as mild grade of poisoning at the entrance of the hospital center is presented. It was managed with antiophidic serum in doses lower than the recommended one, with progression of the symptoms of poisoning. Adjustments in the doses were done and total recovery w...

  7. [Effect of reduction in the Bothrops antivenin dose administrated in patients bitten by the Bothrops snake].

    Science.gov (United States)

    Jorge, M T; Ribeiro, L A

    1994-01-01

    Seven hundred and thirty accidents by Bothrops snake bite attended at Hospital Vital Brazil-Instituto Butantan (HVB-IB) were studied: 411 of in 1983/84 (group A) and 319 in 1986 (group B). All the patients were seen in the Hospital 6 hours after the bite. As an average, all the 411 patients of the group A and the 319 patients of the group B received antivenom capable of neutralizing 215 mg and 117 mg of Bothrops jararaca, respectively. Both groups were similar concerning to: sex and age of the patients; anatomic region bitten; frequency of incision and tourniquet use at the site bite; time in between of the accident and the admission at HVB-IB; classification of B. jararaca either young or adult; frequency of the coagulation impairment. There was no difference in the evolution of the local envenoming manifestations and there were no cases of death. PMID:8061698

  8. Influence of 60Co γ irradiation on fertility of Japanese pine sawyer beetle Monochamus alternatus hope (Coleoptera: Cerambycidae)

    International Nuclear Information System (INIS)

    The fertility of the Japanese pine sawyer beetle Monochamus alternatus (Coleoptera: Cerambycidae) irradiated with 60Co γ-rays was remarkable reduced at the doses of 30Gy, 35Gy and 40Gy, especially at 40Gy. When the non-irradiated females were coupled with the irradiated males first, and then coupled with non-irradiated males, the hatchability and the fertility had little higher but lower than the control. It explained that radiation has certain influence to the female gonad. It also has difference between the hatching rate and the amount of eggs in different match. (authors)

  9. Evaluation of antivenoms in the neutralization of hyperalgesia and edema induced by Bothrops jararaca and Bothrops asper snake venoms

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    Picolo G.

    2002-01-01

    Full Text Available Neutralization of hyperalgesia induced by Bothrops jararaca and B. asper venoms was studied in rats using bothropic antivenom produced at Instituto Butantan (AVIB, 1 ml neutralizes 5 mg B. jararaca venom and polyvalent antivenom produced at Instituto Clodomiro Picado (AVCP, 1 ml neutralizes 2.5 mg B. aspar venom. The intraplantar injection of B. jararaca and B. asper venoms caused hyperalgesia, which peaked 1 and 2 h after injection, respectively. Both venoms also induced edema with a similar time course. When neutralization assays involving the independent injection of venom and antivenom were performed, the hyperalgesia induced by B. jararaca venom was neutralized only when bothropic antivenom was administered iv 15 min before venom injection, whereas edema was neutralized when antivenom was injected 15 min or immediately before venom injection. On the other hand, polyvalent antivenom did not interfere with hyperalgesia or edema induced by B. asper venom, even when administered prior to envenomation. The lack of neutralization of hyperalgesia and edema induced by B. asper venom is not attributable to the absence of neutralizing antibodies in the antivenom, since neutralization was achieved in assays involving preincubation of venom and antivenom. Cross-neutralization of AVCP or AVIB against B. jararaca and B. asper venoms, respectively, was also evaluated. Only bothropic antivenom partially neutralized hyperalgesia induced by B. asper venom in preincubation experiments. The present data suggest that hyperalgesia and edema induced by Bothrops venoms are poorly neutralized by commercial antivenoms even when antibodies are administered immediately after envenomation.

  10. Fatal bothropic snakebite in a horse: a case report

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    NS Silva

    2011-01-01

    Full Text Available The present study reports a snakebite in a horse in the state of Pará, Brazil. At initial evaluation the animal was reluctant to walk and had tachycardia, tachypnea, severe lameness, bleeding on the pastern and swelling around the left hind leg. Blood samples from the bleeding sites, took on the first day, showed leukocytosis and neutrophilia, whereas biochemical values of urea and creatinine were significantly increased. The chosen treatment was snake antivenom, fluid therapy, antibiotics, anti-inflammatory agents and diuretic drugs. On the fourth day of therapy, the hematological values were within normal parameters. There was improvement related to the clinical lameness and swelling of the limb. However, a decrease in water intake and oliguria were observed. On the seventh day the animal died. Necropsy revealed areas of hemorrhagic edema in the left hind limb and ventral abdomen; the kidneys presented equimosis in the capsule, and when cut they were wet. Moreover, the cortex was pale, slightly yellow and the medullary striae had the same aspect. Based on these data, we concluded that the snakebite in the present study was caused by Bothrops spp. and that renal failure contributed to death.

  11. Bothrops lanceolatus bites: guidelines for severity assessment and emergent management.

    Science.gov (United States)

    Resiere, Dabor; Mégarbane, Bruno; Valentino, Ruddy; Mehdaoui, Hossein; Thomas, Laurent

    2010-01-01

    Approximately 20-30 declared snakebite cases occurin Martinique each year. Bothrops lanceolatus, a member of the Crotalidae family, is considered to be the only involved snake. B. lanceolatus, commonly named "Fer-de-Lance", is endemic and only found on this Caribbean island. Envenomation local features include the presence of fang marks, swelling, pain, bleeding from punctures, and ecchymosis. Severe envenomation is associated with multiple systemic thromboses appearing within 48 h of the bite and resulting in cerebral, myocardial or pulmonary infarctions. Diagnosis requires first of all identification of the snake. Coagulation tests are helpful to identify thrombocytopenia or disseminated intravascular coagulation. A clinical score based on 4 grades is helpful to assess envonimation severity. A specific monovalent equine anti-venom (Bothrofav(®), Sanofi-Pasteur, France) to neutralize B. lanceolatus venom is available. Its early administration within 6h from the biting in case of progressive local injures, general signs or coagulation disturbances is effective to prevent severe thrombosis and coagulopathy. Its tolerance is considered to be good. Despite an increasing incidence of bites, no deaths have been recently attributed to B. lanceolatus in Martinique, probably due to the currently recommended strategy of early antivenom administration when required. PMID:22069552

  12. First report of hepatic hematoma after presumed Bothrops envenomation

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    Fernanda Cristina Cunha

    2015-10-01

    Full Text Available ABSTRACTIn Latin America, Bothrops envenomation is responsible for the majority of accidents caused by venomous snakes. Patients usually present local edema, bleeding and coagulopathy. Visceral hemorrhage is extremely rare and considered a challenge for diagnosis and management. We report the first case of hepatic hematoma owing to the bothropic envenomation in a 66-year-old man who was bitten in the left leg. He presented local edema, coagulopathy, and acute kidney injury. Radiological findings suggested hepatic hematoma, with a volume of almost 3 liters. The hepatic hematoma was gradually absorbed without the need for surgical intervention with complete resolution in 8 months.

  13. Ontogenetic variability of Bothrops atrox and Bothrops asper snake venoms from Colombia.

    Science.gov (United States)

    Saldarriaga, Mónica María; Otero, Rafael; Núñez, Vitelbina; Toro, Maria Fabiola; Díaz, Abel; Gutiérrez, José María

    2003-09-15

    The lancehead snakes Bothrops asper and Bothrops atrox inflict 70-90% of the 3000 bites reported every year in Colombia. In this work, the venoms of B. atrox from Meta (Villavicencio, 33 specimens) and B. asper from Antioquia (San Carlos, 45 specimens), all of them born in captivity, were obtained at different ages (0-6 months; 1, 2 and 3-years old) and compared in terms of their pharmacological and immunochemical characteristics. A conspicuous ontogenetic variability was observed in venom samples from both species. Venoms from newborn and juvenile specimens showed higher lethal, hemorrhagic, edema-forming and coagulant activities, whereas venoms from 3-year old specimens showed higher indirect hemolytic, i.e. phospholipase A2 activity, being more significant in the case of B. asper. SDS-polyacrylamide gel electrophoresis of whole venom for both species evidenced a predominance of high mol. mass bands in the venoms from specimens of <1 year of age, with a change towards bands having lower mol. mass as snakes aged. Gel filtration chromatography showed five peaks in the venoms of B. asper of <6 months and in those from 3-year old specimens. Venom of adult specimens showed a higher number of peaks with indirect hemolytic activity than venom of newborn specimens. Polyvalent antivenom produced in Costa Rica recognized all the bands of both venoms from specimens at all ages tested, when assayed by Western blotting. PMID:14505941

  14. Purification of an acidic phospholipase A2 from Bothrops lanceolatus (fer de lance) venom: molecular and enzymatic properties.

    Science.gov (United States)

    de Araújo, A L; Radvanyi, F; Bon, C

    1994-09-01

    The acidic phospholipase A2 from Bothrops lanceolatus venom has been purified by gel filtration on Sephadex G-50 and ion exchange chromatography on CM-cellulose. Analysis by FPLC on Mono-Q column of the purified phospholipase A2 indicated that it is a mixture of several isoenzymes. The two major isoforms consist of a single polypeptide chain with mol. wts of 14,500 and 15,000, which slightly differ in their isoelectric point (4.9 and 5.3) and amino acid composition. However, enzymatic and pharmacological properties of the various isoenzymes are identical. The phospholipase from B. lanceolatus venom is characterized by a progressive increase in the rate of hydrolysis when enzymatic activity is determined with crude egg yolk as substrate in the absence of detergent. This phenomenon, which is not observed with mixed micelles of lecithin-detergent, is not due to the presence of a phospholipase A2 inhibitor in the venom, as previously suggested by several investigators in the case of other Bothrops and Cobra venoms. It is rather a catalytic characteristics of B. lanceolatus venom phospholipase, the enzymatic activity of which depends on the physical state of phospholipids. Bothrops lanceolatus acidic phospholipase A2 is non-toxic. PMID:7801343

  15. Envenomation by Bothrops atrox in a traveler to Manaus, Brazil.

    Science.gov (United States)

    Larréché, Sébastien; Imbert, Patrick; Mornand, Pierre; Andriamanantena, Dina; Mion, Georges; Rapp, Christophe

    2013-01-01

    Snakebites are an infrequent but real risk for travelers. We report a case of envenomation by Bothrops atrox in a traveler to Manaus, Brazil. Rapid administration of specific antivenom prevented the expected systemic disorders. This case gives opportunity to review prevention and first aid measures of snakebites in travelers. PMID:23570867

  16. Brainstem ischemic stroke after to Bothrops atrox snakebite.

    Science.gov (United States)

    Cañas, Carlos A

    2016-09-15

    We report case of a 48 years old woman bitten on her right foot by a Bothrops atrox viper. As a result, she developed a severe coagulopathy which improved with application of polyvalent antivenom. Four days after bite she suffered a devastating brainstem ischemic stroke. Possible pathogenetic mechanisms are discussed. PMID:27527269

  17. Purification and biological effects of a C-type lectin isolated from Bothrops moojeni

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    PSF Barbosa

    2010-01-01

    Full Text Available Snake venom proteins from the C-type lectin family have very distinct biological activities despite their highly conserved primary structure, which is homologous to the carbohydrate recognition region of true C-type lectins. We purified a lectin-like protein (BmLec from Bothrops moojeni venom and investigated its effect on platelet aggregation, insulin secretion, antibacterial activity, and isolated kidney cells. The BmLec was purified using two chromatographic steps: affinity chromatography and reverse phase high performance liquid chromatography (HPLC. BmLec showed a dose-dependent platelet aggregation and significantly decreased the bacterial growth rate in approximately 15%. During scanning electron microscopy, the profile of Xanthomonas axonopodis pv. passiflorae treated with lectin disclosed a high vesiculation and membrane rupture. BmLec induced a strong and significant increase in insulin secretion at 2.8 and 16.7 mM glucose concentrations, and this effect was seen in the presence of EGTA in both experiments. BmLec (10 µg/mL increased the perfusion pressure, renal vascular resistance and urinary flow. The glomerular filtration rate and percentages of sodium, potassium and chloride tubular transport were reduced at 60 minutes of perfusion. Renal alterations caused by BmLec were completely inhibited by indomethacin in all evaluated parameters. In conclusion, the C-type lectin isolated from Bothrops moojeni affected platelet aggregation, insulin secretion, antibacterial activity and isolated kidney function.

  18. Accidente ofídico causado por Bothrops Asper

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    Galofre-Ruiz Mario David

    2013-12-01

    Full Text Available Introduction: The snake bite of the genus Bothrops is an important cause of ophidic accident in Colombia, with high morbidity and mortality. Clinical case: A case of bite by the snake Bothrops Asper, which was classified as mild grade of poisoning at the entrance of the hospital center is presented. It was managed with antiophidic serum in doses lower than the recommended one, with progression of the symptoms of poisoning. Adjustments in the doses were done and total recovery was reached in five days. Conclusions: Antiophidic serums, named also antivenins, are the cornerstone of the treatment to minimize the local tissue damage and the systemic complications. Rev. cienc.biomed. 2013;4(2:353-357

  19. Bothrops lanceolatus Bites: Guidelines for Severity Assessment and Emergent Management

    OpenAIRE

    Resiere, Dabor; Mégarbane, Bruno; Valentino, Ruddy; Mehdaoui, Hossein; Thomas, Laurent

    2010-01-01

    Approximately 20-30 declared snakebite cases occurin Martinique each year. Bothrops lanceolatus, a member of the Crotalidae family, is considered to be the only involved snake. B. lanceolatus, commonly named “Fer-de-Lance”, is endemic and only found on this Caribbean island. Envenomation local features include the presence of fang marks, swelling, pain, bleeding from punctures, and ecchymosis. Severe envenomation is associated with multiple systemic thromboses appearing within 48 h of the bit...

  20. Capacidade reprodutiva de Bothrops jararaca (Serpentes, Viperidae Reproductive capacity of Bothrops jararaca (Serpentes, Viperidae

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    Thélia Rosana Forte Janeiro-Cinquini

    Full Text Available The relationship between female body size and egg number was examined in the snake Bothrops jararaca (Wied, 1924. Some authors have demonstrated that the estimated clutch size may be affected by the inclusion or exclusion of dead neonates or non-developed eggs. The reproductive system of 254 mature females newly arrived from nature was examined and 29.55% (n=75 had eggs in the oviducts in different stages of embryonic development. The presence of atresic eggs was observed in 34.5% of those females. A significant difference was observed in the number of eggs in the right and left oviducts (t=9.48, P<0.001, but not in the number of atresic eggs (t=1.110, P=0.278. The correlation of female body size and egg number was greater when all, the normal and atresic eggs were considered (r=0.63, P<0.001. The exclusion of atresic eggs from the clutch size decreased slightly this correlation (r=0.54, P<0.001.

  1. Therapeutic possibilities of Bothrops jararaca in high dilution

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    Eduardo Costa Gaia Nazareth

    2011-09-01

    Full Text Available Introduction: The knowledge and use of the venom of Bothrops jararaca in high dilutions is still quite limited. One of the important properties is the use of one of its components, bradykinin, for the development of antihypertensive medication known as captopril. Other situations, such as clinical, local and systemic should receive more depth to the composition of Materia Medica related to various medical actions on the man and mammals in general. The systemic action of the bite of this snake, includes hemostasis disorders, culminating as bleeding gums, in addition to sweating, hypertension, and hypothermia. The action includes local pain and swelling with bruising, bleeding and often blistering and tissue necrosis. The action on the immune system, through action on the complement C3 and other complement components may show its possible use in cases of bacterial infections, including mycobacteria, as presented in the study of 1970 Vanessa Birdsey, "Interactions of poisons toxic with the addition, "the journal of Immunology 1971. Today, this poison has a toxicology published by Anibal Melgarejo, "Venomous Animals of Brazil", 2003, which subsidizes the development of study for its use in high dilutions, and a comprehensive study of the biology of the animal itself. Published studies on biomolecular analysis add more details about the relations of the poison and mammals. All these characteristics suggest the use of poison as a homeopathic remedy. Objective: To investigate the therapeutic possibilities in high dilutions of the venom of the snake Bothrops jararaca, expanding its clinical use. Methodology: Methodological description of this poison in contemporary bases including: Origin, physical description chemistry, toxicology, pharmacology and medicine in preparation of high dilution, general action, specific actions on systems or organs, sensations, modalities, concomitants, etiological indications relations main clinics. Results: Defining

  2. Isotopic analysis of Bothrops atrox in Amazonian forest

    Science.gov (United States)

    Martinez, M. G.; Silva, A. M.; Chalkidis, H.; de Oliveira Júnior, R. C.; Camargo, P. B.

    2012-12-01

    The poisoning of snakes is considered a public health problem, especially in populations from rural areas of tropical and subtropical countries. In Brazil, the 26,000 snakebites, 90% are of the genus Bothrops, and Bothrops atrox species predominant in the Amazon region including all the Brazilian Amazon. Research shows that using stable isotopes, we can verify the isotopic composition of tissues of animals that depend mainly on food, water ingested and inhaled gases. For this study, samples taken from Bothrops atrox (B. atrox), in forest using pitfall traps and fall ("Pitt-fall traps with drift fence"). The analyzes were performed by mass spectrometry, where the analytical error is 0.3‰ for carbon and 0.5‰ to nitrogen. The results of the forest animals are significantly different from results of animal vivarium. The average values of the tissues and venoms of snakes of the forest for carbon-13 and nitrogen-15 are: δ13C = -24.68‰ and δ15N = 14.22‰ and mean values of tissue and poisons snakes vivarium (Instituto Butantan) to carbon-13 and nitrogen-15 are δ13C = -20.47‰ and δ15N = 8.36‰, with a significantly different due to different sources of food animals. Based on all results isotopic δ13C and δ15N, we can suggest that changes as the power of the serpent, (nature and captivity), changes occur in relation to diet and environment as the means of the isotopic data are quite distinct, showing that these changes can also cause metabolic changes in the body of the animal itself and the different periods of turnover of each tissue analyzed.

  3. Analgesic Effect of Photobiomodulation on Bothrops Moojeni Venom-Induced Hyperalgesia: A Mechanism Dependent on Neuronal Inhibition, Cytokines and Kinin Receptors Modulation

    Science.gov (United States)

    Oliveira, Victoria Regina da Silva; Toniolo, Elaine Flamia; Feliciano, Regiane dos Santos; da Silva Jr., José Antonio; Zamuner, Stella Regina

    2016-01-01

    Background Envenoming induced by Bothrops snakebites is characterized by drastic local tissue damage that involves an intense inflammatory reaction and local hyperalgesia which are not neutralized by conventional antivenom treatment. Herein, the effectiveness of photobiomodulation to reduce inflammatory hyperalgesia induced by Bothrops moojeni venom (Bmv), as well as the mechanisms involved was investigated. Methodology/Principal Findings Bmv (1 μg) was injected through the intraplantar route in the right hind paw of mice. Mechanical hyperalgesia and allodynia were evaluated by von Frey filaments at different time points after venom injection. Low level laser therapy (LLLT) was applied at the site of Bmv injection at wavelength of red 685 nm with energy density of 2.2 J/cm2 at 30 min and 3 h after venom inoculation. Neuronal activation in the dorsal horn spinal cord was determined by immunohistochemistry of Fos protein and the mRNA expression of IL-6, TNF-α, IL-10, B1 and B2 kinin receptors were evaluated by Real time-PCR 6 h after venom injection. Photobiomodulation reversed Bmv-induced mechanical hyperalgesia and allodynia and decreased Fos expression, induced by Bmv as well as the mRNA levels of IL-6, TNF-α and B1 and B2 kinin receptors. Finally, an increase on IL-10, was observed following LLLT. Conclusion/Significance These data demonstrate that LLLT interferes with mechanisms involved in nociception and hyperalgesia and modulates Bmv-induced nociceptive signal. The use of photobiomodulation in reducing local pain induced by Bothropic venoms should be considered as a novel therapeutic tool for the treatment of local symptoms induced after bothropic snakebites. PMID:27749899

  4. Bothropic antivenom based on monoclonal antibodies, is it possible?

    Science.gov (United States)

    Frauches, Thiago S; Petretski, Jorge H; Arnholdt, Andrea C V; Lasunskaia, Elena B; de Carvalho, Eulógio C Q; Kipnis, Thereza L; da Silva, Wilmar D; Kanashiro, Milton M

    2013-09-01

    Neutralizing monoclonal antibodies against three major toxic components of Bothrops atrox venom were produced and tested. The mAbs against phospholipase A2, hemorrhagic metalloprotease, and thrombin-like enzymes were produced in large amounts and purified with caprylic acid followed by ammonium sulfate precipitation. Purified mAbs were analyzed by SDS-PAGE and their ability to neutralize the respective toxins was tested. Five Swiss mice were injected i.p. with 13.5 mg of pooled mAbs and challenged via s.c. route with venom. Survival rate was recorded for the next 48 h. All mice treated and challenged with venom survived, whereas only one mouse in the control group survived. Bleeding time in mice treated with mAbs was similar to that observed in control mice. Our results show that monoclonal antibodies neutralized the lethal toxicity of Bothrops venom and indicate that there is a reasonable possibility of developing antivenoms based on humanized mAbs to treat victims of venomous animals in the future. PMID:23732123

  5. Comparison of phylogeny, venom composition and neutralization by antivenom in diverse species of bothrops complex.

    Science.gov (United States)

    Sousa, Leijiane F; Nicolau, Carolina A; Peixoto, Pedro S; Bernardoni, Juliana L; Oliveira, Sâmella S; Portes-Junior, José Antonio; Mourão, Rosa Helena V; Lima-dos-Santos, Isa; Sano-Martins, Ida S; Chalkidis, Hipócrates M; Valente, Richard H; Moura-da-Silva, Ana M

    2013-01-01

    In Latin America, Bothrops snakes account for most snake bites in humans, and the recommended treatment is administration of multispecific Bothrops antivenom (SAB--soro antibotrópico). However, Bothrops snakes are very diverse with regard to their venom composition, which raises the issue of which venoms should be used as immunizing antigens for the production of pan-specific Bothrops antivenoms. In this study, we simultaneously compared the composition and reactivity with SAB of venoms collected from six species of snakes, distributed in pairs from three distinct phylogenetic clades: Bothrops, Bothropoides and Rhinocerophis. We also evaluated the neutralization of Bothrops atrox venom, which is the species responsible for most snake bites in the Amazon region, but not included in the immunization antigen mixture used to produce SAB. Using mass spectrometric and chromatographic approaches, we observed a lack of similarity in protein composition between the venoms from closely related snakes and a high similarity between the venoms of phylogenetically more distant snakes, suggesting little connection between taxonomic position and venom composition. P-III snake venom metalloproteinases (SVMPs) are the most antigenic toxins in the venoms of snakes from the Bothrops complex, whereas class P-I SVMPs, snake venom serine proteinases and phospholipases A2 reacted with antibodies in lower levels. Low molecular size toxins, such as disintegrins and bradykinin-potentiating peptides, were poorly antigenic. Toxins from the same protein family showed antigenic cross-reactivity among venoms from different species; SAB was efficient in neutralizing the B. atrox venom major toxins. Thus, we suggest that it is possible to obtain pan-specific effective antivenoms for Bothrops envenomations through immunization with venoms from only a few species of snakes, if these venoms contain protein classes that are representative of all species to which the antivenom is targeted. PMID

  6. Comparison of phylogeny, venom composition and neutralization by antivenom in diverse species of bothrops complex.

    Directory of Open Access Journals (Sweden)

    Leijiane F Sousa

    Full Text Available In Latin America, Bothrops snakes account for most snake bites in humans, and the recommended treatment is administration of multispecific Bothrops antivenom (SAB--soro antibotrópico. However, Bothrops snakes are very diverse with regard to their venom composition, which raises the issue of which venoms should be used as immunizing antigens for the production of pan-specific Bothrops antivenoms. In this study, we simultaneously compared the composition and reactivity with SAB of venoms collected from six species of snakes, distributed in pairs from three distinct phylogenetic clades: Bothrops, Bothropoides and Rhinocerophis. We also evaluated the neutralization of Bothrops atrox venom, which is the species responsible for most snake bites in the Amazon region, but not included in the immunization antigen mixture used to produce SAB. Using mass spectrometric and chromatographic approaches, we observed a lack of similarity in protein composition between the venoms from closely related snakes and a high similarity between the venoms of phylogenetically more distant snakes, suggesting little connection between taxonomic position and venom composition. P-III snake venom metalloproteinases (SVMPs are the most antigenic toxins in the venoms of snakes from the Bothrops complex, whereas class P-I SVMPs, snake venom serine proteinases and phospholipases A2 reacted with antibodies in lower levels. Low molecular size toxins, such as disintegrins and bradykinin-potentiating peptides, were poorly antigenic. Toxins from the same protein family showed antigenic cross-reactivity among venoms from different species; SAB was efficient in neutralizing the B. atrox venom major toxins. Thus, we suggest that it is possible to obtain pan-specific effective antivenoms for Bothrops envenomations through immunization with venoms from only a few species of snakes, if these venoms contain protein classes that are representative of all species to which the antivenom is

  7. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming Associação de heparina e antiveneno: eficácia da neutralização dos venenos de Bothrops atrox e Bothrops erythromelas

    OpenAIRE

    Antônio Luiz R. BOECHAT; Paiva, Cristiano S.; FRANÇA Francisco Oscar; DOS-SANTOS Maria Cristina

    2001-01-01

    Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was no...

  8. In vitro hemolytic activity of Bothrops lanceolatus (fer-de-lance) venom

    OpenAIRE

    LJ Martins; PMF de Araújo; Bon, C.; S. HYSLOP; AL de Araújo

    2009-01-01

    Bothrops lanceolatus venom contains a variety of enzymatic and biological activities. The present work investigated the hemolytic activity of this venom and its phospholipase A2 (PLA2). Bothrops lanceolatus venom (6.7 µg/mL) caused indirect hemolysis of cow, horse, rat and sheep erythrocytes, with horse erythrocytes being the most sensitive; no direct hemolysis was observed. Hemolysis in sheep erythrocytes was concentration-dependent (5-11.7 µg/mL) and markedly attenuated by heating the venom...

  9. ELISA assays for the detection of Bothrops lanceolatus venom in envenomed patient plasmas.

    Science.gov (United States)

    Rodriguez-Acosta, A; Uzcategui, W; Azuaje, R; Giron, M E; Aguilar, I

    1998-01-01

    A double antibody sandwich enzyme linked immunosorbant assay (ELISA) was carried out to detect Bothrops Ianceolatus venom in plasma from envenomed patients at various time intervals (0, 6, 12, 18 and 24 hrs). The test could detect Bothrops lanceolatus levels up to 12 ng/mL of envenomed patient plasmas. Elaboration of an easy, fast and species-diagnostic based on this ELISA technique useful to physicians is discussed. PMID:11845439

  10. Pharmacological characterization and neutralization of the venoms used in the production of Bothropic antivenom in Brazil.

    Science.gov (United States)

    Camey, Kyoko U; Velarde, David T; Sanchez, Eladio F

    2002-05-01

    Some pharmacological effects of the venoms of five different Brazilian Bothrops species and a pool of these venoms (AgB) were quantified. The ability of polyspecific Bothropic antivenom produced at Fundação Ezequiel Dias (FUNED, Brazil) to neutralize the principal toxic and enzymatic activities was studied using in vivo and in vitro assays. The lethality, hemorrhagic, necrotizing, proteolytic, phospholipase, coagulant and fibrinolytic activities were measured for each of these venoms. Comparison of protein electrophoretic patterns showed significant differences such as the presence of common and also unique components. Furthermore, experimental studies revealed differences in their biological properties among individual samples. It was found that the Bothrops antivenom was highly effective in the neutralization of the toxic activities of all venoms tested. In addition, indirect ELISA was used to compare the antigenic cross-reactivity for each of the five Bothrops venoms as well as the venoms of B. atrox, B. leucurus and B. erythromelas which were not included in the antigenic pool (AgB). Therefore, the characterization of several toxic activities of snake venoms is necessary, if toxicity is to be properly evaluated. Results indicate that the Brazilian antivenom prepared at FUNED against Bothrops snakes is effective in neutralizing the main toxic effects of Bothrops venoms. PMID:11821121

  11. Transcriptome analysis of the Amazonian viper Bothrops atrox venom gland using expressed sequence tags (ESTs).

    Science.gov (United States)

    Neiva, Márcia; Arraes, Fabricio B M; de Souza, Jonso Vieira; Rádis-Baptista, Gandhi; Prieto da Silva, Alvaro R B; Walter, Maria Emilia M T; Brigido, Marcelo de Macedo; Yamane, Tetsuo; López-Lozano, Jorge Luiz; Astolfi-Filho, Spartaco

    2009-03-15

    Bothrops atrox is a highly dangerous pit viper in the Brazilian Amazon region. We produced a global catalogue of gene transcripts to identify the main toxin and other protein families present in the B. atrox venom gland. We prepared a directional cDNA library, from which a set of 610 high quality expressed sequence tags (ESTs) were generated by bioinformatics processing. Our data indicated a predominance of transcripts encoding mainly metalloproteinases (59% of the toxins). The expression pattern of the B. atrox venom was similar to Bothrops insularis, Bothrops jararaca and Bothrops jararacussu in terms of toxin type, although some differences were observed. B. atrox showed a higher amount of the PIII class of metalloproteinases which correlates well with the observed intense hemorrhagic action of its toxin. Also, the PLA2 content was the second highest in this sample compared to the other three Bothrops transcriptomes. To our knowledge, this work is the first transcriptome analysis of an Amazonian rain forest pit viper and it will contribute to the body of knowledge regarding the gene diversity of the venom gland of members of the Bothrops genus. Moreover, our results can be used for future studies with other snake species from the Amazon region to investigate differences in gene patterns or phylogenetic relationships. PMID:19708221

  12. Inflammation induced by Bothrops asper venom: release of proinflammatory cytokines and eicosanoids, and role of adhesion molecules in leukocyte infiltration.

    Science.gov (United States)

    Zamuner, Stella Regina; Zuliani, Juliana Pavan; Fernandes, Cristina Maria; Gutiérrez, José Maria; de Fátima Pereira Teixeira, Catarina

    2005-12-01

    Bothrops asper venom (BaV) causes systemic and local effects characterized by an acute inflammatory reaction with accumulation of leukocytes and release of endogenous mediators. In this study, the effects of BaV on the release of the cytokines IL-1, IL-6 and TNF-alpha and the eicosanoids LTB4 and TXA2 in the peritoneal cavity of mice were analyzed. We also investigated the participation of beta2 integrin chain, l-selectin, LFA-1, ICAM-1 and PECAM-1 adhesion molecules in the BaV-induced leukocyte accumulation. Levels of proinflammatory cytokines IL-6 and TNF-alpha, as well as eicosanoids LTB4 and TXA2 were significantly increased after BaV injection (250 microg/kg), whereas no increment in IL-1 was observed. Anti-mouse l-selectin, LFA-1, ICAM-1, PECAM-1 and beta2 integrin chain monoclonal antibodies resulted in a reduction of neutrophil accumulation induced by BaV injection compared with isotype-matched control injected animals. These data suggest that BaV is able to induce the activation of leukocytes and endothelium to express adhesion molecules involved in the recruitment of neutrophils into the inflammed site. Furthermore, these results showed that BaV induces the release of cytokines and eicosanoids in the local of the venom injection; these inflammatory mediators may be important for the initiation and amplification of the inflammatory reaction characteristic from Bothrops sp envenomation. PMID:16198389

  13. Lance-headed viper (Bothrops moojeni bite wounding the eye Acidente botrópico (Bothrops moojeni com ferimento ocular

    Directory of Open Access Journals (Sweden)

    Érico Otaviano Brandão

    1993-08-01

    Full Text Available A 5-year-old girl was bitten in her left eye by a lance-headed viper identified as Bothrops moojeni, measuring 115 cm of length. There was severe facial swelling and left exophthalmus, and enucleation of the eye was necessary. The patient apparently had mild systemic envenoming, but local inflammatory signs and histological evidence of necrosis suggest that both the mechanical trauma and the local action of the venom had a role in the genesis of the eye lesion. It is arguable if the loss of the eye could be prevented even if the antivenom was administered earlier.Uma menina de 5 anos de idade foi picada no olho esquerdo por uma serpente da espécie Bothrops moojeni, que media 115 cm de comprimento. Houve intenso edema facial e ex-oftalmia; enucleação do olho foi necessária. A paciente aparentemente apresentou um envenenamento sistêmico leve, mas os sinais inflamatórios locais e a evidência histológica de necrose sugerem que tanto o traumatismo mecânico como a ação local do veneno tiveram um papel na gênese da lesão ocular. É questionável se o prognóstico da lesão ocular teria sido diferente se o antiveneno tivesse sido administrado em dose adequada mais precocemente.

  14. Screening of Superior Strain of Beauveria bassiana Parasitized on Monochamus alternatus%松墨天牛寄生白僵菌的优良菌株筛选

    Institute of Scientific and Technical Information of China (English)

    马良进; 杨毅; 张立钦

    2006-01-01

    对21个球孢白僵茵(Beauveria bassiana)茵株的生长、产孢能力、抗逆性等生物学性状以及对松墨天牛(M. alternatus)4龄幼虫的毒力进行了全面的比较研究.综合各菌株的生物学特性和对天牛幼虫的毒力指标,最终得出防治松墨天牛幼虫最优良菌株是来自浙江象山的Bxs和来自江苏的B5菌株,其中Bxs在生长指标上略占优势,B5在抗性指标上略占优势.

  15. Clinical and immunological aspects of envenomations by Bothrops snakes

    Directory of Open Access Journals (Sweden)

    KPO Luna

    2011-01-01

    Full Text Available Accidents caused by snakes, especially in tropical and subtropical countries, still constitute a serious public health problem due to the lack of knowledge of health professionals and the precariousness of health systems in the regions where most accidents occur. Snake venoms contain a range of molecules that may provoke local swelling, pain, renal and respiratory insufficiencies. The study of the effects of each molecule on humans can help the development of complementary therapy. Similarly, the knowledge of clinical aspects of envenomations provides a better identification and implementation of appropriate treatment. In addition, to understand Bothrops envenomations and improve the therapeutic strategy, it is necessary to understand and study the role of important inflammatory mediators, particularly nitric oxide (NO, cytokines and the complement system.

  16. Occipital infarction revealed by quadranopsia following snakebite by Bothrops lanceolatus.

    Science.gov (United States)

    Merle, Harold; Donnio, Angélique; Ayeboua, Lucas; Plumelle, Yves; Smadja, Didier; Thomas, Laurent

    2005-09-01

    We report a case of snakebite in which envenomation was manifested through impairment of the visual field. The patient, a 46-year-old man, was bitten on the right thumb by Bothrops lanceolatus. Treatment with a specific equine antivenom (Bothrofav) was administered one hour after the bite. With the exception of fang marks, the results of a clinical examination, particularly the neurologic component, were normal. The day after the bite, the patient developed an inferior left lateral homonymous quadranopsia with macular epargne. T2 magnetic resonance imaging showed a right occipital infarction. His condition improved clinically and biologically. This observation of snakebite is the first in which envenomation was accompanied exclusively by an impairment of the visual field. Envenomation by B. lanceolatus is distinct in its incidence of significant thrombotic complications at a distance from the site of the bite. PMID:16172485

  17. Acidente ofídico em ovinos causado por Bothrops jararaca Snake bite accident in sheep by Bothrops jararaca

    Directory of Open Access Journals (Sweden)

    Carlos Hubinger Tokarnia

    2008-12-01

    Full Text Available Relata-se a ocorrência de um acidente ofídico em um rebanho de ovinos, mestiços da raça Bergamascia, no município de Miguel Pereira, Estado do Rio de Janeiro. Quatro ovelhas adultas, com cerca 35 kg de peso, foram picados na cabeça por um exemplar de Bothrops jararaca dentro de curto espaço de tempo. Os animais apresentaram a cabeça muito tumefeita, mas o estado geral era bom. Um ovino foi eutanasiado. Verificou-se que a tumefação da cabeça era causada por hemorragia embaixo da pele, que se estendia ao pescoço. Histologicamente foi verificada na face, hemorragia maciça, principalmente no tecido subcutâneo, com infiltração de hemácias no tecido muscular adjacente, que em áreas onde era mais intensa, provocou afastamento entre as fibras musculares, algumas com aspecto hialinizado. Discute-se a causa das diferenças sobre a ocorrência de edema/hemorragia e necrose no envenenamento causado pelas diversas serpentes do gênero Bothrops nas diferentes espécies animais.The occurrence of a snake bite accident in a herd of mixed Bergamascia sheep in the county of Miguel Pereira, State of Rio de Janeiro, is reported. Four adult sheep were bitten in the head by Bothrops jararaca within a short period of time. Their head was severely swollen, but the general state of them was good. One of the sheep was euthanasied. Masssive hemorrhages were found beneath the skin, which extended to the neck. Histologically, the subcutaneous hemorrhages infiltrated the adjacent muscles. In areas where the hemorrhages were more intense they caused separation of muscle fibers, some of which had a hyaline aspect. Differences in the occurrence of edema/hemorraghes and necrosis in snake bite accidents, caused by various snake species in different animal species, are discussed.

  18. Cloning, Characterization and Anti-Inflammatory Properties of Bothrops jararaca Snake Antithrombin.

    Science.gov (United States)

    Morais-Zani, Karen de; Grego, Kathleen F; Torquato, Ricardo J S; Silva, Caroline S; Tanaka, Aparecida S; Tanaka-Azevedo, Anita M

    2015-01-01

    Antithrombin inhibits blood coagulation through the interaction with serine proteases in both intrinsic and extrinsic pathways. In addition, antithrombin also shows anti-inflammatory properties, which are independent of its effects on coagulation. This work shows for the first time the cloning and sequencing of antithrombin from a snake species. This predicted protein is composed by 430 amino acids and presents about 64.5% sequence identity to human antithrombin. Biacore experiments revealed that the binding affinity of Bothrops jararaca snake antithrombin to heparin was ~30 times higher than that of human antithrombin. Furthermore, Bothrops jararaca antithrombin is more effective in preventing acute inflammation induced by carrageenan when compared to human antithrombin. Hence, the results showed herein suggest that Bothrops jararaca antithrombin can play a key role in the control of acute inflammation and that this molecule might be used as a pharmacological tool and as a prototype for drug development. PMID:25687119

  19. Neuromuscular action of Bothrops lanceolatus (Fer de lance) venom and a caseinolytic fraction.

    Science.gov (United States)

    Lôbo de Araújo, Albetiza; Donato, José Luiz; Leite, Gildo Bernardo; Prado-Franceschi, Júlia; Fontana, Marcos Dias; Bon, Cassian; Rodrigues Simioni, Léa

    2002-09-01

    A protein capable of inducing neuromuscular blockade in avian preparations and of depolarizing mouse diaphragm muscle was isolated from Bothrops lanceolatus venom using gel filtration and ion-exchange chromatography. The purified protein was a single chain polypeptide with an estimated molecular mass of 27.5 kDa by SDS-PAGE and had caseinolytic activity (13.3 units/mg), but no phospholipase A(2). B.lanceolatus venom (50 micro g/ml) and the caseinolytic protein (20 micro g/ml) produced contracture and progressive irreversible blockade (50% in 25+/-5 min (SEM) and 45+/-15 min, respectively), in indirectly stimulated chick biventer cervicis preparations. The contractile responses to acetylcholine (ACh; 37 and 74 micro M, n=6) were inhibited by venom and the caseinolytic protein, whereas those to potassium (13.4mM, n=6) were not. Membrane resting potential measurements in mouse hemidiaphragm preparations showed that B.lanceolatus venom and the purified protein caused depolarization which was prevented by D-tubocurarine (14.6mM). The venom produced a slight increase in the amplitude and frequency of miniature end-plate potentials, but this effect was not seen with the purified fraction. These results suggest that the purified protein acts exclusively post-synaptically. PMID:12220713

  20. Hemorrhagic stroke in children caused by Bothrops marajoensis envenoming: a case report.

    Science.gov (United States)

    de Oliveira Pardal, Pedro Pereira; Pinheiro, Augusto Cezar Jennings da Silva; Silva, Cristiane Tarcis Cunha; Santos, Paulo Roberto Silva Garcez; Gadelha, Maria Apolônia da Costa

    2015-01-01

    According to the World Health Organization, snakebites are considered neglected diseases. Bothrops, the genus most frequently implicated in envenomations in Brazil, includes the species B. marajoensis Hoge, 1966, part of the complex B. atrox, which is found in the savannas of Marajó Island, Pará state, Brazil, a region that presents scarce epidemiological data. This work reports the first case of hemorrhagic stroke in a child, attributed to delayed medical care after snakebite envenoming by Bothrops marajoensis in Anajás city, Marajó Island, Pará, Brazil, which led to permanent hemiplegia as a sequela. PMID:26672486

  1. Reproductive Biology of Bothrops erythromelas from the Brazilian Caatinga

    Directory of Open Access Journals (Sweden)

    Verônica Alberto Barros

    2014-01-01

    Full Text Available The reproductive biology of Bothrops erythromelas, a small pit viper from the Caatinga, a semiarid biome in Brazil, is described based on analysis of individuals deposited in zoological collections. Males are smaller and also attain sexual maturity at a smaller size than females. Female reproductive cycle is seasonal with an extended period of secondary vitellogenesis and births occurring in a restricted period from late spring to early summer. Sperm storage in females may probably occur in infundibular tubular glands and uterine muscular twisting (UMT, which is a polymorphic condition within B. erythromelas. Seasonal spermatogenesis in males is variable with some intraspecific variation regarding the male reproductive stage per season. Most males are reproductively active during spring/summer and reproductively quiescent during autumn/winter, although some individuals vary (e.g., show testicular spermatogenesis and active sexual segment of the kidneys (SSK during winter. The SSK could be identified in every male. Most males showed highly hypertrophied SSK in spring/summer and moderately hypertrophied SSK in autumn/winter. The ampulla ductus deferentis was observed and histochemical reactions were conducted. We discuss the probable influence of the unique environmental conditions of the Caatinga region and phylogenetic inertia in the reproductive patterns of this snake species.

  2. 松褐天牛入侵新疆的风险分析%Risk Analysis of Monochamus alternatus in Xinjiang

    Institute of Scientific and Technical Information of China (English)

    张新平; 焦淑萍; 张静文; 岳朝阳

    2013-01-01

    Qualitative and quantitative analyses were carried out on the risk of the invasion of Monochamus alternatus to Xinjiang from the following aspects: distribution of the pest,potential harmfulness,economic importance of host plants,possibility of the pest transmission,and the difficulty of management. Comprehensive evaluation was conducted on the risk of the invasion of the pest to Xinjiang. The results showed that M. alternatus was a highly dangerous forest pest in Xinjiang. Its host plants were widely distributed in the region. In order to prevent the diffusion and extension of the pest,measures should be taken from the strengthening of quarantine to prevent the invasion of the pest via the transportation channels of nursery stocks, branches, logs, boards. Onnce it was imported, the situation should be monitored immediately and relative control measures would be taken.%根据蒋青提出的有害生物危险性的定量分析方法及相关文献,对松褐天牛在我国的分布、潜在的危害性、寄主植物的经济重要性、传播扩散的可能性以及危险性的管理难度等几个方面进行定性和定量分析,并对松褐天牛入侵新疆的危险性作出综合评价.结果表明,松褐天牛在新疆属于高度危险的森林有害生物.松褐天牛寄主植物在新疆的分布范围较广,为防止松褐天牛入侵我区扩散蔓延,应加强检疫关,严防其通过苗木、枝桠、原木、板材等的调运而传入,一旦传入,必须及时进行监测及相关的防治处理.

  3. Determination of inorganic elements in blood of mice immunized with Bothrops Snake venom using XRF and NAA

    Science.gov (United States)

    Lopes da Silva, L. F. F.; Zamboni, C. B.; Bahovschi, V.; Metairon, S.; Suzuki, M. F.; Sant'Anna, O. A.; Rizzutto, M. A.

    2015-07-01

    In this work, mice genetically modified [HIII line] were immunized against different Bothrops snake venoms to produce anti-Bothrops serum (antivenom). The Neutron Activation Analysis (NAA) and Energy Dispersive X-Ray Fluorescence (EDXRF) techniques were used to evaluate Ca and Fe concentrations in blood of these immunized mice in order to establish a potential correlation between both phenotypes: antibody response and blood constituents after Bothrops venom administration. The results were compared with the control group (mice not immunized) and with human being estimative. These data are important for clinical screening of patients submitted to immunological therapy as well as the understanding of the envenoming mechanisms.

  4. NAA TECHNIQUE FOR CLINICAL INVESTIGATION OF MICE IMMUNIZED WITH BOTHROP VENOM

    International Nuclear Information System (INIS)

    In the present study Neutron Activation Analysis (NAA) technique was used to determine sodium concentration in whole blood of mice immunized with Bothrops venom. With this value it was possible to perform clinical investigation in this animal model using whole blood.

  5. Antigenic cross-reactivity and immunogenicity of Bothrops venoms from snakes of the Amazon region.

    Science.gov (United States)

    Furtado, Maria de Fátima D; Cardoso, Silvia Travaglia; Soares, Oscar Espellet; Pereira, Aparecida Pietro; Fernandes, Daniel Silva; Tambourgi, Denise Vilarinho; Sant'Anna, Osvaldo Augusto

    2010-04-01

    Snakebites are still a critical public health problem in developing countries or isolated areas. In Brazil, the North Region has a high distribution coefficient worsened by the significant number of eventually unreported cases, due to difficulties in access to health services, to the natural geographic barriers and the vast territory. In the Rio Negro area, the species Bothrops atrox, Bothrops brazili, Lachesis muta muta and Bothriopsis taeniata are thought to be the major species responsible for snakebites. The aim of this study was to qualitatively and quantitatively determine the antigenic cross-reactivity and expression of toxins and the immunogenicity of Bothrops venom species of the Amazon and to evaluate the general efficacy of the therapeutic sera. The in vivo assays demonstrated that the defibrinating activity of B. taeniata venom was absent but that the lethal and hemorrhagic properties were more intense than in the B. atrox venom. The results evidence venom variability among the two B. atrox populations from two distinct Amazonian regions, which may reveal a subjacent speciation process. The results point to new aspects that may guide the improvement of anti-Bothropic therapeutic serum. PMID:20036275

  6. Natural resistance of opossum (Didelphis marsupialis) to the mapanare (Bothrops lanceolatus) snake venom.

    Science.gov (United States)

    Pifano, F; Aguilar, I; Giron, M E; Gamboa, N; Rodriguez-Acosta, A

    1993-01-01

    The inactivation of local and general effects of the Mapanare (Bothrops lanceolatus) venom by Opossum's (Didelphis marsupialis) serum fractions was tested using an in vivo assay and an in vitro preincubation experiment. A serum fraction of the Opossum serum has been obtained by immunochemical purification. It is only present in opossum's protective opossum serum fraction (F-0.1). PMID:8186456

  7. Purification and characterization of a hemorrhagic metalloproteinase from Bothrops lanceolatus (Fer-de-lance) snake venom.

    Science.gov (United States)

    Stroka, Alessandra; Donato, José L; Bon, Cassian; Hyslop, Stephen; de Araújo, Albetiza Lôbo

    2005-03-15

    Bothrops snake venoms contain metalloproteinases that contribute to the local effects seen after envenoming. In this work, a hemorrhagic metalloproteinase (BlaH1) was purified from the venom of the snake Bothrops lanceolatus by a combination of gel filtration, affinity (metal chelating) and hydrophobic interaction chromatographies. The hemorrhagin was homogeneous by SDS-PAGE and had a molecular mass of 28 kDa that was unaltered by treatment with beta-mercaptoethanol. BlaH1 gave a single band in immunoelectrophoresis and immunoblotting using commercial bothropic antivenom. BlaH1 had hemorrhagic, caseinolytic, fibrinogenolytic, collagenolytic and elastinolytic activities, but no phospholipase A(2) activity. The hemorrhagic and caseinolytic activities were inhibited by EDTA, indicating that they were metal ion-dependent. In contrast, aprotinin, benzamidine and PMSF did not affect these activities. The caseinolytic activity of BlaH1 had a pH optimum of 8.0 and was stable in solution at up to 40 degrees C; activity was completely lost at > or =70 degrees C. The hemorrhagic activity was neutralized by commercial bothropic antivenom. These properties suggest that this new hemorrhagin belongs to class P-I snake venom metalloproteinases. PMID:15733562

  8. Toxicity of Bothrops sp snake venoms from Ecuador and preclinical assessment of the neutralizing efficacy of a polyspecific antivenom from Costa Rica.

    Science.gov (United States)

    Laines, Johana; Segura, Álvaro; Villalta, Mauren; Herrera, María; Vargas, Mariángela; Alvarez, Gladys; Gutiérrez, José María; León, Guillermo

    2014-09-01

    The toxicological profile of the venoms of the snakes Bothrops asper and Bothrops atrox from Ecuador was investigated, together with the venom of a population of B. asper formerly classified as 'Bothrops xanthogrammus'. The three venoms exerted lethal, hemorrhagic, myotoxic, coagulant and defibrinogenating effects, in agreement with the characteristic toxicological profile of Bothrops sp venoms. A polyspecific antivenom (bothropic-crotalic-lachesic) manufactured in Costa Rica was assessed for its preclinical efficacy against the toxic activities of these Ecuadorian venoms. Antivenom was effective in the neutralization of the five activities tested in the three venoms. These observations are in agreement with previous reports on the extensive cross-reactivity and paraspecific neutralization of antivenoms manufactured in Latin America against the venoms of Bothrops sp snakes. PMID:24950051

  9. The distribution and elimination of Bothrops erythromelas venom labeled with 131 I after intravenous injection in mice

    International Nuclear Information System (INIS)

    Pharmacokinetic studies can be used to study the systemic effects of snake venoms and to develop standard serotherapy protocols for envenomation. Bothrops erythromelas is probably responsible for most of the snakebite in Pernambuco. The objective of this study was to investigate the pharmacokinetics of B. erythromelas venom (BeV) in mice, and to evaluate the efficacy of bothropic antivenom. BeV showed bicompartmental distribution in the blood of the experimental animals. (author)

  10. The distribution and elimination of Bothrops erythromelas venom labeled with {sup 131} I after intravenous injection in mice

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, M.L. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Zoologia]. E-mail: rocha@cascavel.uefs.br

    1999-07-01

    Pharmacokinetic studies can be used to study the systemic effects of snake venoms and to develop standard serotherapy protocols for envenomation. Bothrops erythromelas is probably responsible for most of the snakebite in Pernambuco. The objective of this study was to investigate the pharmacokinetics of B. erythromelas venom (BeV) in mice, and to evaluate the efficacy of bothropic antivenom. BeV showed bicompartmental distribution in the blood of the experimental animals. (author)

  11. Effects of irradiated Bothropstoxin-1 and Bothrops jararacussu crude venom on the immune system

    International Nuclear Information System (INIS)

    Ionizing radiation has been successfully employed to modify the immunological properties of biomolecules and has been proven to be a powerful tool to attenuate snake venoms toxicity without affecting and even increasing their immunogenic properties. Very promising results were obtained when crude animal venoms, as well as isolated toxins, were treated with 60Co gamma rays, yielding toxoids with good immunogenicity, however, little is known about the modifications that irradiated molecules undergo and even less about the immunological response that such antigens elicit. At the present work, we have evaluated the effects on immune system of B10.PL and BALB/c mice of Bothrops jararacussu crude venom and isolated bothropstoxin-1 (Bthx-1), before and after gamma radiation exposition. According to our data, irradiation process promoted structural modifications on both isolated toxin and crude venom, characterized by higher molecular weight protein (aggregates and oligomers) formation. Irradiated samples were immunogenic and the antibodies elicited by them were able to recognize the native toxin in ELISA. These results indicate that irradiation of toxic proteins can promote significant modifications in their structures, but still retain many of the original antigenic and immunological properties. Also, our data indicate that the irradiated protein induced higher titers of IgG2b, suggesting that Th1 cells were predominantly involved. Results from Western blot assay showed that antibodies raised against irradiated bothropstoxin-1 recognize both native isolated toxin or crude venom. Cytotoxicity assay showed that irradiated toxin and crude venom were less toxic than their native counterpart. Thus, the viability of the macrophages cultured in the presence of irradiated Bthx-1 or crude venom was higher if compared with their native forms. LDH Assay showed that irradiated Bthx-1 promotes less muscular damage than the native form. Our data confirm a potential use of ionizing

  12. A transcriptomic view of the proteome variability of newborn and adult Bothrops jararaca snake venoms.

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    André Zelanis

    Full Text Available BACKGROUND: Snake bite is a neglected public health problem in communities in rural areas of several countries. Bothrops jararaca causes many snake bites in Brazil and previous studies have demonstrated that the pharmacological activities displayed by its venom undergo a significant ontogenetic shift. Similarly, the venom proteome of B. jararaca exhibits a considerable variation upon neonate to adult transition, which is associated with changes in diet from ectothermic prey in early life to endothermic prey in adulthood. Moreover, it has been shown that the Brazilian commercial antibothropic antivenom, which is produced by immunization with adult venom, is less effective in neutralizing newborn venom effects. On the other hand, venom gland transcripts of newborn snakes are poorly known since all transcriptomic studies have been carried out using mRNA from adult specimens. METHODS/PRINCIPAL FINDINGS: Here we analyzed venom gland cDNA libraries of newborn and adult B. jararaca in order to evaluate whether the variability demonstrated for its venom proteome and pharmacological activities was correlated with differences in the structure of toxin transcripts. The analysis revealed that the variability in B. jararaca venom gland transcriptomes is quantitative, as illustrated by the very high content of metalloproteinases in the newborn venom glands. Moreover, the variability is also characterized by the structural diversity of SVMP precursors found in newborn and adult transcriptomes. In the adult transcriptome, however, the content of metalloproteinase precursors considerably diminishes and the number of transcripts of serine proteinases, C-type lectins and bradykinin-potentiating peptides increase. Moreover, the comparison of the content of ESTs encoding toxins in adult male and female venom glands showed some gender-related differences. CONCLUSIONS/SIGNIFICANCE: We demonstrate a substantial shift in toxin transcripts upon snake development and a

  13. Snake venomics of Bothrops punctatus, a semiarboreal pitviper species from Antioquia, Colombia

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    Maritza Fernández Culma

    2014-01-01

    Full Text Available Bothrops punctatus is an endangered, semi-arboreal pitviper species distributed in Panamá, Colombia, and Ecuador, whose venom is poorly characterized. In the present work, the protein composition of this venom was profiled using the ‘snake venomics’ analytical strategy. Decomplexation of the crude venom by RP-HPLC and SDS-PAGE, followed by tandem mass spectrometry of tryptic digests, showed that it consists of proteins assigned to at least nine snake toxin families. Metalloproteinases are predominant in this secretion (41.5% of the total proteins, followed by C-type lectin/lectin-like proteins (16.7%, bradykinin-potentiating peptides (10.7%, phospholipases A2 (93%, serine proteinases (5.4%, disintegrins (38%, L-amino acid oxidases (3.1%, vascular endothelial growth factors (17%, and cysteine-rich secretory proteins (1.2%. Altogether, 6.6% of the proteins were not identified. In vitro, the venom exhibited proteolytic, phospholipase A2, and L-amino acid oxidase activities, as well as angiotensin-converting enzyme (ACE-inhibitory activity, in agreement with the obtained proteomic profile. Cytotoxic activity on murine C2C12 myoblasts was negative, suggesting that the majority of venom phospholipases A2 likely belong to the acidic type, which often lack major toxic effects. The protein composition of B. punctatus venom shows a good correlation with toxic activities here and previously reported, and adds further data in support of the wide diversity of strategies that have evolved in snake venoms to subdue prey, as increasingly being revealed by proteomic analyses.

  14. Cell adhesion molecules involved in the leukocyte recruitment induced by venom of the snake Bothrops jararaca

    Directory of Open Access Journals (Sweden)

    Stella R. Zamuner

    2002-01-01

    Full Text Available It has been shown that Bothrops jararaca venom (BjV induces a significant leukocyte accumulation, mainly neutrophils, at the local of tissue damage. Therefore, the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1, LECAM-1, CD18, leukocyte function-associated antigen-1 (LFA-1 and platelet endothelial cell adhesion molecule-1 (PECAM-1 on the BjV-induced neutrophil accumulation and the correlation with release of LTB4, TXA2, tumor necrosis factor-α, interleukin (IL-1 and IL-6 have been investigated. Anti-mouse LECAM-1, LFA-1, ICAM-1 and PECAM-1 monoclonal antibody injection resulted in a reduction of 42%, 80%, 66% and 67%, respectively, of neutrophil accumulation induced by BjV (250 μg/kg, intraperitoneal injection in male mice compared with isotype-matched control injected animals. The anti-mouse CD18 monoclonal antibody had no significant effect on venom-induced neutrophil accumulation. Concentrations of LTB4, TXA2, IL-6 and TNF-α were significant increased in the peritoneal exudates of animals injected with venom, whereas no increment in IL-1 was detected. This results suggest that ICAM-1, LECAM-1, LFA-1 and PECAM-1, but not CD18, adhesion molecules are involved in the recruitment of neutrophils into the inflammatory site induced by BjV. This is the first in vivo evidence that snake venom is able to up-regulate the expression of adhesion molecules by both leukocytes and endothelial cells. This venom effect may be indirect, probably through the release of the inflammatory mediators evidenced in the present study.

  15. Radioiodination and biodistribution of Leucurolysin-B isolated from Bothrops Leucurus in mice bearing Ehrlich

    Energy Technology Data Exchange (ETDEWEB)

    Gabriel, L.M.; Soares, M.A.; Bicalho, M.S.; Santos, R.G. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: marcellaaraugio@yahoo.com.br, e-mail: mbs@cdtn.br, e-mail: santosr@cdtn.br; Sanchez, E.O.F.; Silva, S.G. [Ezequiel Dias Foundation, Belo Horizonte, MG (Brazil)], e-mail: silea@funed.mg.gov.br, e-mail: eladio@funed.mg.gov.br

    2009-07-01

    Integrins are family of heterodimeric cell surface adhesion receptors able to recognize and bind to proteins in the extracellular matrix (ECM). This recognition is mainly through the RGD domain present in both the cell surface as the protein in the ECM. Various integrins have been identified as regulators of tumor progression. The RGD domain is also found in some snake venoms named disintegrins. Disintegrins inhibit cell-matrix and a cell-cell interaction mediated by integrin and has been shown that these proteins are able to inhibit metastasis in processes dependent on integrin. The disintegrin-like (ECD), as well as RGD-disintegrin are also able to bind to cell surface integrins and inhibit their adherence to the natural ligands. Leucurolysin-B (Leuc-B) is a metalloproteinase class P-III isolated from Bothrops leucurus (BLV) and possesses a disintegrin-like domain (ECD). The goals of this work were to synthesize a radioactive probe analog to Leuc-B using radioiodine {sup 125}I and evaluate the interaction of {sup 125}I-Leuc-B in tumor cells through the study of biodistribution in animals bearing Ehrlich tumor.125I-Leuc-B was synthesized using lactoperoxidase with high yield (90%) and specific activity of 1.2x10-7Bq/mmol. It was observed that {sup 125}I-Leuc-B had very fast clearance from the blood stream (T1/2= 0.01 h). Tumor uptake of 125I-Leuc-B gradually increased up to (2 min) and remained for a quite long period. The tumor/normal tissue uptake ratios of {sup 125}I-Leuc-B were 1.77 (tumor/normal paw) and 8.44 tumor/skeletal muscle. The results suggest that {sup 125}I-Leuc- B may constitute a good template for development of a tool for detection of solid tumors. (author)

  16. Distribution of 131 I- labeled Bothrops erythromelas venom in mice

    International Nuclear Information System (INIS)

    Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with 131 I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%.Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc) with 0.3 ml (2.35 x 105 cpm/mouse) of 131 I-venom (N = 42), 131 -albumin or 131 I (controls, N = 28 each). Thirty minutes and 1,3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% Na Cl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption int he skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach ( 3 rd h),small intestine (3 rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3 rd h postinjection (1.98%).There was a low binding of labeled venom in the liver (B. erythromelas venom does not specifically target most internal organs. That is, the systemic effects of envenomation ar mainly due to an indirect action. (author)

  17. Tissue damage caused by Bothrops sp envenoming evaluated by magnetic resonance imaging (MRI)

    Energy Technology Data Exchange (ETDEWEB)

    Fonseca, M.G. [Faculdade do Norte Paulista, Bebedouro, SP (Brazil); Matias, M.R.C. [UNESP, Botucatu, SP (Brazil). Hospital Universitario. Unidade de Ressonancia Magnetica; Yamashita, S.; Morceli, J. [UNESP, Botucatu, SP (Brazil). Faculdade de Medicina. Dept. de Doencas Tropicais e Diagnostico por Imagem; Barraviera, B. [Universidade Estadual Paulista (UNESP), Botucatu, SP (Brazil). Centro de Estudos de Venenos e Animais Venenosos (CEVAP)]. E-mail: marilucefonseca@hotmail.com

    2002-07-01

    The objective of this clinical study was to evaluate local tissue damage caused by Bothrops sp envenoming in relation to lesion type and damaged tissues using magnetic resonance imaging (MRI). Fifteen patients bitten by Bothrops snakes were treated at the Emergency Unit of the Tropical Diseases Unit at the University Hospital, Botucatu School of Medicine, UNESP, Sao Paulo State, Brazil. After receiving specific sero therapy, the patients were submitted to MR of the bite site. T 1 spin-echo MRI were obtained revealing the following lesions: edema (n=9), edema associated with hemorrhage (n=5), and hemorrhage (n=1). Peri muscular areas (n=6) and subcutaneous tissues (n=5) were the most affected, followed by muscular tissues (n=4). It is important to mention that MRI did not show myonecrosis of the bite site, a widely reported finding in anatomical and histopathological experimental studies. (author)

  18. Detection of an antibothropic fraction in opossum (Didelphis marsupialis) milk that neutralizes Bothrops jararaca venom.

    Science.gov (United States)

    Jurgilas, P B; Neves-Ferreira, A G; Domont, G B; Moussatché, H; Perales, J

    1999-01-01

    An antibothropic fraction (ABF) from Didelphis marsupialis (opossum) serum, which is responsible for the neutralization of Bothrops jararaca venom was isolated by Perales et al. [Perales, J., Moussatché, H., Marangoni, S., Oliveira, B. and Domont, G. B. (1994). Isolation and partial characterization of an antibothropic complex from the serum of South American Didelphidae. Toxicon 32, 1237-1249]. The aim of this work was to verify the presence of this factor in opossum's milk, which could represent an additional protection for the neonatal opossum against bothropic venoms. An active milk fraction was isolated and showed similar physicochemical, structural, antigenic and biological properties when compared to ABF, indicating that they are probably the same protein. PMID:9920488

  19. Snake Venomics and Antivenomics of Bothrops diporus, a Medically Important Pitviper in Northeastern Argentina.

    Science.gov (United States)

    Gay, Carolina; Sanz, Libia; Calvete, Juan J; Pla, Davinia

    2016-01-01

    Snake species within genus Bothrops are responsible for more than 80% of the snakebites occurring in South America. The species that cause most envenomings in Argentina, B. diporus, is widely distributed throughout the country, but principally found in the Northeast, the region with the highest rates of snakebites. The venom proteome of this medically relevant snake was unveiled using a venomic approach. It comprises toxins belonging to fourteen protein families, being dominated by PI- and PIII-SVMPs, PLA₂ molecules, BPP-like peptides, L-amino acid oxidase and serine proteinases. This toxin profile largely explains the characteristic pathophysiological effects of bothropic snakebites observed in patients envenomed by B. diporus. Antivenomic analysis of the SAB antivenom (Instituto Vital Brazil) against the venom of B. diporus showed that this pentabothropic antivenom efficiently recognized all the venom proteins and exhibited poor affinity towards the small peptide (BPPs and tripeptide inhibitors of PIII-SVMPs) components of the venom. PMID:26712790

  20. Snake venomics and antivenomics of Bothrops colombiensis, a medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela: Contributing to its taxonomy and snakebite management.

    Science.gov (United States)

    Calvete, Juan J; Borges, Adolfo; Segura, Alvaro; Flores-Díaz, Marietta; Alape-Girón, Alberto; Gutiérrez, José María; Diez, Nardy; De Sousa, Leonardo; Kiriakos, Demetrio; Sánchez, Eladio; Faks, José G; Escolano, José; Sanz, Libia

    2009-03-01

    The taxonomic status of the medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela, which has been classified as Bothrops colombiensis, remains incertae cedis. To help resolving this question, the venom proteome of B. colombiensis was characterized by reverse-phase HPLC fractionation followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venom contained proteins belonging to 8 types of families. PI Zn(2+)-metalloproteinases and K49 PLA(2) molecules comprise over 65% of the venom proteins. Other venom protein families comprised PIII Zn(2+)-metalloproteinases (11.3%), D49 PLA(2)s (10.2%), l-amino acid oxidase (5.7%), the medium-sized disintegrin colombistatin (5.6%), serine proteinases (1%), bradykinin-potentiating peptides (0.8%), a DC-fragment (0.5%), and a CRISP protein (0.1%). A comparison of the venom proteomes of B. colombiensis and B. atrox did not support the suggested synonymy between these two species. The closest homologues to B. colombiensis venom proteins appeared to be toxins from B. asper. A rough estimation of the similarity between the venoms of B. colombiensis and B. asper indicated that these species share approximately 65-70% of their venom proteomes. The close kinship of B. colombiensis and B. asper points at the ancestor of B. colombiensis as the founding Central American B. asper ancestor. This finding may be relevant for reconstructing the natural history and cladogenesis of Bothrops. Further, the virtually indistinguishable immunological crossreactivity of a Venezuelan ABC antiserum (raised against a mixture of B. colombiensis and Crotalus durissus cumanensis venoms) and the Costa Rican ICP polyvalent antivenom (generated against a mixture of B. asper, Crotalus simus, and Lachesis stenophrys venoms) towards the venoms of B. colombiensis and B. asper, supports this

  1. Daño renal en ratas inducido por veneno de Bothrops neuwiedii diporus de Argentina Renal injury in rats poisoned by venom of Bothrops neuwiedii diporus from Argentina

    Directory of Open Access Journals (Sweden)

    Patricia Koscinczuk

    2004-08-01

    Full Text Available La insuficiencia renal aguda es una de las complicaciones sistémicas más frecuentes después de un accidente ofídico. En este estudio se evalúan los efectos que el veneno de Bothrops neuwiedii diporus produce en el riñón de ratas machos de la cepa Wistar. La histopatología permitió comprobar el desarrollo de necrosis tubular aguda; las lesiones iniciales se observaron a las 3 horas de la inoculación de una dosis de 700 µg del veneno, observándose en corteza renal congestión y degeneración granulohialina de las células epiteliales tubulares, acompañadas de dilatación y cilindros hialinos en la luz tubular. A las 24 horas se presentó necrosis tubular aguda en una superficie extensa de la corteza sin daño de la membrana basal tubular. Las lesiones de degeneración turbia de células epiteliales tubulares, dilatación de la luz tubular y cilindros hialinos se mantuvieron presentes hasta las 4 semanas post-inoculación. Si bien los parámetros de la bioquímica clínica asociados con insuficiencia renal aguda aumentaron a las 6 horas de la administración del veneno (urea: 1.10±0.22 g/dl; creatinina: 19.60±1.51mg/dl, a la semana descendieron a valores normales. Las densidades urinarias, en cambio, a la semana se mantuvieron más bajas que lo normal, 1.005 ± 0.001 (pAcute renal failure is one of the systemic complications that can be found in bothropic accidents. In this study the effects on male Wistar rats induced by the venom of Bothrops neuwiedii diporus were evaluated. The histopathology revealed acute tubular necrosis, lesions firstly were observed 3 hours post inoculation of 700 µg of venom. Cortical kidney congestion and granulohialin degeneration of tubular epithelial cells were observed, these lesions achieved a maximum at 24 hours after inoculation. Tubular epithelial hidropic degeneration and dilatation of tubular lumen with hyalin casts were present inclusive up to 4 weeks after inoculation. Biochemical parameter

  2. A C-Type Lectin from Bothrops jararacussu Venom Disrupts Staphylococcal Biofilms

    OpenAIRE

    Raphael Contelli Klein; Mary Hellen Fabres-Klein; Leandro Licursi de Oliveira; Renato Neves Feio; François Malouin; Andréa de Oliveira Barros Ribon

    2015-01-01

    Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chro...

  3. Cell adhesion molecules involved in the leukocyte recruitment induced by venom of the snake Bothrops jararaca

    OpenAIRE

    Catarina F. P. Teixeira; Stella R. Zamuner

    2002-01-01

    It has been shown that Bothrops jararaca venom (BjV) induces a significant leukocyte accumulation, mainly neutrophils, at the local of tissue damage. Therefore, the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1), LECAM-1, CD18, leukocyte function-associated antigen-1 (LFA-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1) on the BjV-induced neutrophil accumulation and the correlation with release of LTB4, TXA2, tumor necrosis factor-alpha, interleukin (I...

  4. Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox

    OpenAIRE

    Sousa Marcelo V; Paba Jaime; López-Lozano Jorge L; Shevchenko Andrej; Shevchenko Anna; Guércio Rafael AP; Ricart Carlos AO

    2006-01-01

    Abstract Background Bothrops atrox is responsible for the majority of snakebite accidents in the Brazilian Amazon region. Previous studies have demonstrated that the biological and pharmacological activities of B. atrox venom alter with the age of the animal. Here, we present a comparative proteome analysis of B. atrox venom collected from specimens of three different stages of maturation: juveniles, sub-adults and adults. Results Optimized conditions for two-dimensional gel electrophoresis (...

  5. Neutralizing capacity of a new monovalent anti-Bothrops atrox antivenom: comparison with two commercial antivenoms

    OpenAIRE

    Otero, R; V. Núñez; Gutiérrez, J M; Robles, A; R. Estrada; R.G. Osorio; G. Del-Valle; R. Valderrama; Giraldo, C. A.

    1997-01-01

    Three horse-derived antivenoms were tested for their ability to neutralize lethal, hemorrhagic, edema-forming, defibrinating and myotoxic activities induced by the venom of Bothrops atrox from Antioquia and Chocó (Colombia). The following antivenoms were used: a) polyvalent (crotaline) antivenom produced by Instituto Clodomiro Picado (Costa Rica), b) monovalent antibothropic antivenom produced by Instituto Nacional de Salud-INS (Bogotá), and c) a new monovalent anti-B. atrox antivenom produce...

  6. Immunological assessment of mice hyperimmunized with native and Cobalt-60-irradiated Bothrops venoms

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira Junior, R.S.; Meira, D.A. [UNESP, Botucatu, SP (Brazil). Dept. de Doencas Tropicais; Barraviera, B. [UNESP, Botucatu, SP (Brazil). Center for the Study of Venoms and Venomous Animals - CEVAP; Nascimento, N.; Alves, J.B. [Instituto de Pesquisas Energeticas (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Martinez, J.C. [UNESP, Botucatu, SP (Brazil). Escola de Engenharia]. E-mail: rseabra@cevap.org.br

    2005-10-15

    ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated ({sup 60}Co) venoms of Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by in vivo challenge, and the side effects were assessed. The animals immunization with one LD50 of each venom occurred on days 1, 15, 21, 30, and 45, when blood samples were collected; challenges happened on the 60th day. Results showed that ELISA was efficient in evaluating, accompanying and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after {sup 60} Co-irradiation. The sera produced with native venom showed neutralizing potency and capacity similar to those of the sera produced with irradiated venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom, however, necrosis and death occurred in animals inoculated with native venom. (author)

  7. Immunological assessment of mice hyperimmunized with native and Cobalt-60-irradiated Bothrops venoms

    International Nuclear Information System (INIS)

    ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated (60Co) venoms of Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by in vivo challenge, and the side effects were assessed. The animals immunization with one LD50 of each venom occurred on days 1, 15, 21, 30, and 45, when blood samples were collected; challenges happened on the 60th day. Results showed that ELISA was efficient in evaluating, accompanying and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60 Co-irradiation. The sera produced with native venom showed neutralizing potency and capacity similar to those of the sera produced with irradiated venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom, however, necrosis and death occurred in animals inoculated with native venom. (author)

  8. Nomenclatural instability in the venomous snakes of the Bothrops complex: Implications in toxinology and public health.

    Science.gov (United States)

    Carrasco, Paola Andrea; Venegas, Pablo Javier; Chaparro, Juan Carlos; Scrocchi, Gustavo José

    2016-09-01

    Since nomenclature is intended to reflect the evolutionary history of organisms, advances in our understanding of historical relationships may lead to changes in classification, and thus potentially in taxonomic instability. An unstable nomenclature for medically important animals like venomous snakes is of concern, and its implications in venom/antivenom research and snakebite treatment have been extensively discussed since the 90´s. The taxonomy of the pitvipers of the Bothrops complex has been historically problematic and different genus-level rearrangements were proposed to rectify the long-standing paraphyly of the group. Here we review the toxinological literature on the Bothrops complex to estimate the impact of recent proposals of classification in non-systematic research. This assessment revealed moderate levels of nomenclatural instability in the last five years, and the recurrence of some practices discussed in previous studies regarding the use of classifications and the information provided about the origin of venom samples. We briefly comment on a few examples and the implications of different proposals of classifications for the Bothrops complex. The aim of this review is to contribute to the reduction of adverse effects of current taxonomic instability in a group of medical importance in the Americas. PMID:27242040

  9. Duvernoy's gland secretion of Philodryas olfersii and Philodryas patagoniensis (Colubridae): neutralization of local and systemic effects by commercial bothropic antivenom (Bothrops genus).

    Science.gov (United States)

    Rocha, Marisa M Teixeira; Paixão-Cavalcante, Danielle; Tambourgi, Denise V; Furtado, Maria de Fátima D

    2006-01-01

    Colubrids involved in human envenomation in Brazil are mainly from the genera Helicops, Oxyrhopus, Thamnodynastes and Philodryas. There is a relatively large number of clinical descriptions involving the Xenodontinae snakes, Philodryas olfersii and Philodryas patagoniensis, in human accidents. The most common manifestations of envenomation are local pain, swelling, erythema and ecchymosis and regional lymphadenopathy with normal coagulation. The aims of this study were to characterize the biochemical and biological properties of P. olfersii and P. patagoniensis venoms, and to investigate their immunological cross-reactivities by using both specific antisera and anti-Bothrops sp serum used for human serum therapy in Brazil, in neutralizing the lethal and hemorrhagic effects of these venoms. We show here that P. olfersii e P. patagoniensis venoms present proteolytic and haemorrhagic activities but are devoid of phospholipase A2 activity. Haemorrhage and lethality induced by P. olfersii and P. patagoniensis are associated with metal-dependent proteinases, since EDTA could block these toxic activities. P. olfersii and P. patagoniensis venoms were immunogenic and the antisera produced were able to recognize several bands in P. olfersii, P. patagoniensis venoms in Bothrops jararaca venom. PMID:16360723

  10. A tripeptide isolated from Bothrops atrox venom has neuroprotective and neurotrophic effects on a cellular model of Parkinson's disease.

    Science.gov (United States)

    Martins, N M; Santos, N A G; Sartim, M A; Cintra, A C O; Sampaio, S V; Santos, A C

    2015-06-25

    Parkinson's disease (PD) is the second most common neurodegenerative disorder; however, there is no treatment able to prevent the loss of dopaminergic neurons or its consequences. Trophic factors such as NGF and BDNF has positive effects on different disorders of the brain, including neurodegeneration. Additionally, studies have suggested the use of venom peptides as a therapeutic strategy for neurological disorders. Therefore, in the present study, we investigated the neuroprotective activity of a peptide isolated from Bothrops atrox venom and its trophic ability by using a cellular model of dopaminergic neurotoxicity induced by 1-methyl-4-phenylpyridinium (MPP(+)) in PC12 cells. We showed that it decreased the activities of the apoptotic proteases caspase-9 (mitochondrial) and caspase-3 (executor) and increased cell viability and proliferation in this model. Additionally, it increased neuritogenesis in non-treated PC12 cells (neuronal model) as well as in PC12 cells treated with the dopaminergic neurotoxin. The amino acid sequence of the peptide was identified as Glutamic acid-Valine-Tryptophan (Glu-Val-Trp). These findings suggest that this tripeptide has the potential to protect against the dopaminergic neurons loss and that trophic stimulation of neuroplasticity might be involved in its mechanism of neuroprotection. PMID:25868679

  11. Acidente vascular cerebral hemorrágico associado à acidente ofídico por serpente do gênero bothrops: relato de caso Hemorrhagic stroke related to snakebite by bothrops genus: a case report

    Directory of Open Access Journals (Sweden)

    Amanda Silva Machado

    2010-10-01

    Full Text Available Este trabalho tem como objetivo relatar um caso de acidente vascular cerebral hemorrágico, associado à acidente ofídico por serpente do gênero bothrops e hipertensão arterial sistêmica grave. Apesar do ofidismo botrópico ser frequente no Estado do Pará, tais associações são incomuns, necessitando de uma abordagem especializada e precoce, visando menores complicações.This research reports a clinical case of hemorrhagic stroke due to envenomation by bothrops snakebite associated with severe hypertension. Although bothrops snakebites are frequent in the State of Pará, such associations are uncommon, requiring specialized and early management to avoid severe complications.

  12. [Pulmonary embolism and disseminated intravascular coagulation after being bitten by a Bothrops lanceolatus snake. Apropos of a case].

    Science.gov (United States)

    Estrade, G; Garnier, D; Bernasconi, F; Donatien, Y

    1989-11-01

    The authors report the case of a Bothrops lanceolatus snake bite complicated by severe pulmonary embolism a few hours after admission. This thromboembolic complication developed despite heparin therapy and was followed by disseminated intravascular coagulation (DIC). Vascular thrombosis and pulmonary embolism are rare after Bothrops lanceolatus snake bite as patients are usually hypocoagulable due to DIC. In this case, the thromboembolism was probably caused by the procoagulant effect of the thrombin-like enzymes of the snake venom which may have been injected directly into the vein of a young woman taking a contraceptive pill. A specific antivenin which has recently become available fort treatment may decrease the complications of Bothrops lanceolatus snake bite. PMID:2514645

  13. Clinical indicators of envenoming and serum levels of venom antigens in patients bitten by Bothrops lanceolatus in Martinique. Research Group on Snake Bites in Martinique.

    Science.gov (United States)

    Bucher, B; Canonge, D; Thomas, L; Tyburn, B; Robbe-Vincent, A; Choumet, V; Bon, C; Ketterlé, J; Lang, J

    1997-01-01

    An enzyme-linked immunosorbent assay was developed to measure venom antigen levels in the serum of 40 patients bitten by Bothrops lanceolatus. The grading system used for the severity of envenomation (grades 1 to 4, minor to major) was predominantly based on the presence of local signs. Serum venom levels increased with the grade of severity (P or = 15 ng/mL were observed in all patients with progressive aggravation of swelling despite the use of early antivenom therapy. No venom was detectable in blood samples taken after completion of serotherapy. All patients recovered. These results confirm the efficacy of both the clinical severity scoring system used and the therapeutic regimen. PMID:9196765

  14. Evidence of caspase-mediated apoptosis induced by l-amino acid oxidase isolated from Bothrops atrox snake venom.

    Science.gov (United States)

    Alves, Raquel Melo; Antonucci, Gilmara Ausech; Paiva, Helder Henrique; Cintra, Adélia Cristina Oliveira; Franco, João José; Mendonça-Franqueiro, Elaine Paula; Dorta, Daniel Junqueira; Giglio, José Roberto; Rosa, José César; Fuly, André Lopes; Dias-Baruffi, Marcelo; Soares, Andreimar Martins; Sampaio, Suely Vilela

    2008-12-01

    The aim of this work was to investigate the involvement of caspases in apoptosis induced by l-amino acid oxidase isolated from Bothrops atrox snake venom. The isolation of LAAO involved three chromatographic steps: molecular exclusion on a G-75 column; ion exchange column by HPLC and affinity chromatography on a Lentil Lectin column. SDS-PAGE was used to confirm the expected high purity level of BatroxLAAO. It is a glycoprotein with 12% sugar and an acidic character, as confirmed by its amino acid composition, rich in "Asp and Glu" residues. It displays high specificity toward hydrophobic l-amino acids. The N-terminal amino acid sequence and internal peptide sequences showed close structural homology to other snake venom LAAOs. This enzyme induces in vitro platelet aggregation, which may be due to H2O2 production by LAAOs, since the addition of catalase completely inhibited the aggregation effect. It also showed cytotoxicity towards several cancer cell lines: HL60, Jurkat, B16F10 and PC12. The cytotoxicity activity was abolished by catalase. A fluorescence microscopy evaluation revealed a significant increase in the apoptotic index of these cells after BatroxLAAO treatment. This observation was confirmed by phosphatidyl serine exposure and activation of caspases. BatroxLAAO is a protein with various biological functions that can be involved in envenomation. Further investigations of its function will contribute to toxicology advances. PMID:18804547

  15. Fatal diffuse thrombotic microangiopathy after a bite by the "Fer-de-Lance" pit viper (Bothrops lanceolatus) of Martinique.

    Science.gov (United States)

    Malbranque, Stéphane; Piercecchi-Marti, Marie Dominique; Thomas, Laurent; Barbey, Christophe; Courcier, Dominique; Bucher, Bernard; Ridarch, Alex; Smadja, Didier; Warrell, David A

    2008-06-01

    In Martinique, a man bitten two days earlier by a pit viper (Bothrops lanceolatus) was hospitalized with impaired consciousness and tetraplegia. Investigations confirmed cerebral and myocardial infarctions. Resolving thrombocytopenia was associated with virtually normal blood prothrombin time/activated partial thromboplastin time but increasing hyperfibrinogenemia. Despite specific antivenom treatment, he developed fatal left ventricular failure six days after the bite. At autopsy, multiple cerebral, myocardial and mesenteric infarctions were found. Rupture of mitral chordae tendinae was the likely cause of death. Histopathologic examination showed multi-focal thrombotic microangiopathy with intimal-medial dissection by thrombi extending from foci of endothelial damage in small cerebral, myocardial, pulmonary, mesenteric, and interlobular renal arteries and arterioles. These findings were the causes of infarctions. There was intense angiogenesis in organizing cerebral infarcts. Immunohistochemical analysis showed platelet aggregates and endothelial cells within microthrombi. Viperidae venoms contain vascular endothelial toxins, notably metalloproteinase hemorrhagins, but von Willebrand factor activators or vascular endothelial growth factor-type factors are more likely to have been implicated in this case. PMID:18541759

  16. Human neutrophil migration and activation by BJcuL, a galactose binding lectin purified from Bothrops jararacussu venom

    Directory of Open Access Journals (Sweden)

    Fernandes Luiz

    2011-01-01

    Full Text Available Abstract Background Neutrophil migration to an inflamed site constitutes the first line of the innate immune response against invading microorganisms. Given the crucial role of endogenous lectins in neutrophil mobilization and activation, lectins from exogenous sources have often been considered as putative modulators of leukocyte function. Lectins purified from snake venom have been described as galactoside ligands that induce erythrocyte agglutination and platelet aggregation. This study evaluated human neutrophil migration and activation by C-type lectin BJcuL purified from Bothrops jararacussu venom. Results Utilizing fluorescence microscopy, we observed that biotinylated-BJcuL was evenly distributed on the neutrophil surface, selectively inhibited by D-galactose. Lectin was able to induce modification in the neutrophil morphology in a spherical shape for a polarized observed by optical microscopy and exposure to BJcuL in a Boyden chamber assay resulted in cell migration. After 30 minutes of incubation with BJcuL we found enhanced neutrophil functions, such as respiratory burst, zymozan phagocytosis and an increase in lissosomal volume. In addition, BJcuL delays late apoptosis neutrophils. Conclusion These results demonstrate that BJcuL can be implicated in a wide variety of immunological functions including first-line defense against pathogens, cell trafficking and induction of the innate immune response since lectin was capable of inducing potent neutrophil activation.

  17. The role of TLR2 in the acute inflammatory response induced by Bothrops atrox snake venom.

    Science.gov (United States)

    Moreira, Vanessa; Teixeira, Catarina; Borges da Silva, Henrique; D'Império Lima, Maria Regina; Dos-Santos, Maria Cristina

    2016-08-01

    Envenomation by snakes of the species Bothrops atrox induces local and systemic effects. Local effects include drastic tissue damage and a marked inflammatory response as a result of the synthesis and release of a variety of protein and lipid mediators. Toll-like receptor (TLR) signaling pathways can play an important role in this response, leading to synthesis of these inflammatory mediators. This study investigated the influence of TLR2 on the acute inflammatory response induced by Bothrops atrox venom. Wild-type C57BL/6 mice (WT) and TLR2 gene knockout mice (TLR2(-/-)) were injected with Bothrops atrox venom (BaV), and the following responses to the venom were assessed in peritoneal exudate: leukocyte accumulation; release of mediators, including CCL-2, IL-10, IL-1β, IL-6 and LTB4; protein expression of COX-1 and COX-2; and quantification of their products PGE2 and TXA2. After injection with BaV, the TLR2(-/-) mice (TLR2(-/-)BaV) had higher levels of IL-6 and CCL-2 than WT animals kept under the same conditions (WTBaV), together with an accumulation of polymorphonuclear leukocytes (PMNs), inhibition of IL-1β and LTB4 and reduced mononuclear leukocyte influx. However, no significant differences in COX-2 protein expression or PGE2, TXA2 and IL-10 production between the TLR2(-/-)BaV and WTBav animals were observed. Together, these results indicate that the signaling pathway activated by TLR2 acts by modulating the induced inflammatory response to BaV through the direct action of venom-associated molecular patterns (VAMPs) or indirectly by forming damage-associated molecular patterns (DAMPs) and that this may have important therapeutic implications. PMID:27109323

  18. Pharmacological characterization of the rat paw edema induced by Bothrops lanceolatus (Fer de lance) venom.

    Science.gov (United States)

    de Faria L; Antunes, E; Bon, C; de Araújo, A L

    2001-06-01

    The inflammatory response induced by Bothrops lanceolatus venom (BLV) in the rat hind-paw was studied measuring paw edema. Non-heated BLV (75microg/paw) caused a marked paw edema accompanied by intense haemorrhage whereas heated venom (97 degrees C, 30s; 12.5-100microg/paw) produced a dose- and time-dependent non-haemorrhagic edema. The response with heated BLV was maximal within 15min disappearing over 24h. Heated venom was then routinely used at the dose of 75microg/paw. The prostacyclin analogue iloprost (0.1microg/paw) potentiated by 125% the venom-induced edema. The histamine H(1) receptor antagonist mepyramine (6mg/kg) or the serotonin/histamine receptor antagonist cyproheptadine (6mg/kg) partially inhibited BLV-induced edema whereas the combination of both compounds virtually abolished the edema. The lipoxygenase inhibitor BWA4C (10mg/kg), but not the cyclooxygenase inhibitor indomethacin (10mg/kg), significantly inhibited the edema (35% reduction; P<0.05). Dexamethasone (1mg/kg) also markedly (P<0.001) reduced venom-induced edema. The bradykinin B(2) receptor antagonist Hoe 140 (0.6mg/kg) reduced by 30% (P<0.05) the venom induced edema, whereas the angiotensin-converting enzyme inhibitor captopril (300microg/paw) potentiated by 42% (P<0.05) the edema. Bothrops lanceolatus antivenon (anti-BLV) reduced by 28% (P<0.05) the venom-induced edema while intravenous administration of antivenom failed to affect the edema. In conclusion, BLV-induced rat paw edema involves mast cell degranulation causing local release of histamine and serotonin, a phenomenon mediated mainly by kinins and lipoxygenase metabolites. Additionally, the use of a specific Bothrops lanceolatus antivenom, given subplantarily or intravenously, revealed to be little effective to prevent BLV-induced edema. PMID:11137542

  19. Inactivation and fragmentation of lectin from Bothrops leucurus snake venom by gamma irradiation

    Science.gov (United States)

    Nunes, E. S.; Souza, M. A. A.; Vaz, A. F. M.; Coelho, L. C. B. B.; Aguiar, J. S.; Silva, T. G.; Guarnieri, M. C.; Melo, A. M. M. A.; Oliva, M. L. V.; Correia, M. T. S.

    2012-04-01

    Gamma radiation alters the molecular structure of biomolecules and is able to mitigate the action of snake venoms and their isolated toxins. The effect of γ-radiation on the folding of Bothrops lecurus venom lectin was measured by a hemagglutinating assay, intrinsic and bis-ANS fluorescence. Intrinsic and bis-ANS fluorescence analyses indicated that irradiation caused unfolding followed by aggregation of the lectin. Our results suggest that irradiation can lead to significant changes in the protein structure, which may promote the loss of its binding property and toxic action.

  20. Variação entre filhotes de representantes do complexo Bothrops newied (Serpentes, Viperidae, Crotalinae

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    Vinícius Xavier

    2002-01-01

    Full Text Available External morphological characters of 141 young specimens (69 males and 72 femalesof the Bothrops newied complex were analyzed. Regression analysis was used in the study of morphometric characters and principal components analysis was used in the study of meristic and qualitative characters. Sexual dimorphism was confirmed in the meristic and morphometric characters. Males showed higher counts of subcaudals and longer tails. Females showed eventually higher number of ventrals and dorsal rows, and larger heads. Six different drawing patterns were diagnosed and can indicate the existence of different species. Ontogenetic variation was described.

  1. Two related thrombin-like enzymes present in Bothrops atrox venom

    OpenAIRE

    J.H. Petretski; Kanashiro, M; Silva, C. P.; E.W. Alves; Kipnis, T L

    2000-01-01

    This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical Km on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs) raised against the purified enzyme forms recognized ...

  2. Estudo bacteriológico de abscessos causados por picada de serpentes do genero bothrops Bacterial flora of abscesses following Bothrops snakebite

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    João guimarães De Andrade

    1989-12-01

    Full Text Available Foi analisada a flora bacteriana de 99 abscessos causados por picadas de serpentes do gênero Bothrops, correspondendo a 61,1% dos casos que ocorreram em 1030 acidentes ofídicos atendidos no Hospital de Doenças Tropiciais (HDT de Goiânia, no período de janeiro de 1984 a abril de 1988. O exsudato dos abscessos foi estudado através de bacterioscopia, cultura e testes de sensibilidade para aeróbios. Os bacilos Gram negativos foram isolados em maior frequência, destacando-se a Morganella morganii, Escherichia coli e Providencia sp presentes respectivamente em 44,4%, 20,2% e 13,1% das amostras. Esta flora aeróbica foi semelhante à encontrada na cavidade oral e no veneno das serpentes em outros estudos, nos quais predominaram Morganella morganii. Foi sugerido o uso do cloranfenicol no tratamento dos abscessos que não respondam à simples drenagem, face à grande sensibilidade destes microorganismos demonstrada nos testes "in vitro".The bacterial flora of 99 cases of abscesses following Bothrops snakebite were analysed. They corresponded to 61.1% of all snakebite abs cesses observed in 1030 patients attending the Hospital de Doenças Tropicais de Goiânia in Goiás, Brazil, from January 1984 to April 1988. An exsudate sample of each abscess was examined by Gram stain, culture and susceptibility tests. The Gram negative bacillis, Morganella morganii, Escherichia coli and Providencia sp were the most frequent bacterias isolated. They were identified in 44.4%, 20.2% and 13.1% of the samples respectively. This flora was similar to those described in snake mouth and venom by other researchers. Based on the results of the susceptibility tests the authors suggested the use of chloramphenicol for the treatment of those abscesses which do not respond to simple drainage.

  3. Fatores associados à incoagulabilidade sangüínea no envenenamento por serpentes do gênero Bothrops Risk factors associated with coagulation abnormalities in Bothrops envenoming

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    Ricardo Borges de Oliveira

    2003-12-01

    Full Text Available Com o objetivo de conhecer fatores associados à incoagulabilidade sangüínea no envenenamento botrópico, foram obtidas informações de 2.991 prontuários médicos de pacientes atendidos no Instituto Butantan de 1981 a 1990. Associaram-se positivamente à incoagulabilidade sangüínea (p0,05: horário do acidente; presença de presa recém-deglutida no tubo digestivo da serpente; sexo e idade do paciente; ocorrência de bolha, necrose, abscesso e incisão local, amputação, insuficiência renal e óbito. Pode-se concluir que, embora a incoagulabilidade sangüínea apresente associação com manifestações precoces do envenenamento, não tem boa associação com a evolução clínica do paciente.This study aimed at assessing, in the envenoming by Bothrops, factors that are associated with blood incoagulability. Information was obtained from the charts of 2,991 patients admitted to Instituto Butantan, from 1981 to 1990. Factors positively associated with blood incoagulability (p0.05 were: time of the bite; presence of recently swallowed prey in the snake gut; gender and age of the patient; blister, necrosis, and abscess at the bite site; occurrence of amputation, renal failure and death; presence of an incision at the bite site. We conclude that although blood incoagulability is associated with early manifestations of Bothrops envenoming, it is not associated with the clinical outcome.

  4. Snake venomics of the Lesser Antillean pit vipers Bothrops caribbaeus and Bothrops lanceolatus: correlation with toxicological activities and immunoreactivity of a heterologous antivenom.

    Science.gov (United States)

    Gutiérrez, José María; Sanz, Libia; Escolano, José; Fernández, Julián; Lomonte, Bruno; Angulo, Yamileth; Rucavado, Alexandra; Warrell, David A; Calvete, Juan J

    2008-10-01

    The venom proteomes of the snakes Bothrops caribbaeus and Bothrops lanceolatus, endemic to the Lesser Antillean islands of Saint Lucia and Martinique, respectively, were characterized by reverse-phase HPLC fractionation, followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venoms contain proteins belonging to seven ( B. caribbaeus) and five ( B. lanceolatus) types of toxins. B. caribbaeus and B. lanceolatus venoms contain phospholipases A 2, serine proteinases, l-amino acid oxidases and zinc-dependent metalloproteinases, whereas a long disintegrin, DC-fragments and a CRISP molecule were present only in the venom of B. caribbaeus, and a C-type lectin-like molecule was characterized in the venom of B. lanceolatus. Compositional differences between venoms among closely related species from different geographic regions may be due to evolutionary environmental pressure acting on isolated populations. The venoms of these two species differed in the composition and the relative abundance of their component toxins, but they exhibited similar toxicological and enzymatic profiles in mice, characterized by lethal, hemorrhagic, edema-forming, phospholipase A 2 and proteolytic activities. The venoms of B. caribbaeus and B. lanceolatus are devoid of coagulant and defibrinogenating effects and induce only mild local myotoxicity in mice. The characteristic thrombotic effect described in human envenomings by these species was not reproduced in the mouse model. The toxicological profile observed is consistent with the abundance of metalloproteinases, PLA 2s and serine proteinases in the venoms. A polyvalent (Crotalinae) antivenom produced in Costa Rica was able to immunodeplete approximately 80% of the proteins from both B. caribbaeus and B. lanceolatus venoms, and was effective in neutralizing the lethal, hemorrhagic, phospholipase

  5. Biological and immunological properties of the venom of Bothrops alcatraz, an endemic species of pitviper from Brazil.

    Science.gov (United States)

    Furtado, M F D

    2005-06-01

    Bothrops alcatraz is a new pitviper species derived from the Bothrops jararaca group, whose natural habitat is situated in Alcatrazes Archipelago, a group of marine islands near São Paulo State coast in Brazil. Herein, the biological and biochemical properties of venoms of four adult specimens of B. alcatraz were examined comparatively to a reference pool of Bothrops jararaca venom. Both venoms showed similar activities and electrophoretic patterns, but B. alcatraz venom showed three protein bands of molecular masses of 97, 80 and 38 kDa that were not present in B. jararaca reference venom. The i.p. median lethal dose of B. alcatraz venom ranged from 5.1 to 6.6 mg/kg, while it was 1.5 mg/kg for B. jararaca venom. The minimum hemorrhagic dose of B. jararaca venom was 0.63, whereas 2.28 mug/mouse for B. alcatraz venom. In contrast, B. alcatraz venom was more potent in regard to procoagulant and proteolytic activities. These differences were supported by western blotting and neutralization tests, employing commercial bothropic antivenom, which showed that hemorrhagic and lethal activities of B. alcatraz venom were less effectively inhibited than B. jararaca venom. Such results evidence that B. alcatraz shows quantitative and qualitative differences in venom composition in comparison with its B. jararaca relatives, which might represent an optimization of venom towards a specialized diet. PMID:16002343

  6. Local inflammatory events induced by Bothrops atrox snake venom and the release of distinct classes of inflammatory mediators.

    Science.gov (United States)

    Moreira, Vanessa; Dos-Santos, Maria Cristina; Nascimento, Neide Galvão; Borges da Silva, Henrique; Fernandes, Cristina Maria; D'Império Lima, Maria Regina; Teixeira, Catarina

    2012-07-01

    Bothrops atrox is responsible for most accidents involving snakes in the Brazilian Amazon and its venom induces serious systemic and local effects. The local effects are not neutralized effectively by commercial antivenoms, resulting in serious sequelae in individuals bitten by this species. This study investigates the local inflammatory events induced in mice by B. atrox venom (BaV), such as vascular permeability, leukocyte influx and the release of important inflammatory mediators such as cytokines, eicosanoids and the chemokine CCL-2, at the injection site. The effect of BaV on cyclooxygenase (COX-1 and COX-2) expression was also investigated. The results showed that intraperitoneal (i.p.) injection of BaV promoted a rapid and significant increase in vascular permeability, which reached a peak 1 h after venom administration. Furthermore, BaV caused leukocyte infiltration into the peritoneal cavity between 1 and 8 h after i.p. injection, with mononuclear leukocytes (MNs) predominating in the first 4 h, and polymorphonuclear leukocytes (PMNs) in the last 4 h. Increased protein expression of COX-2, but not of COX-1, was detected in leukocytes recruited in the first and fourth hours after injection of BaV. The venom caused the release of eicosanoids PGD₂, PGE₂, TXA₂ and LTB₄, cytokines TNF-α, IL-6, IL-10 and IL-12p70, but not IFN-γ, and chemokine CCL-2 at different times. The results show that BaV is able to induce an early increase in vascular permeability and a leukocyte influx to the injection site consisting mainly of MNs initially and PMNs during the later stages. These phenomena are associated with the production of cytokines, the chemokine CCL-2 and eicosanoids derived from COX-1 and COX-2. PMID:22465491

  7. Effects of Co60 gamma radiation on the immunogenic and antigenic properties of Bothrops jararacussu venom

    International Nuclear Information System (INIS)

    Ionizing radiation has been successfully employed to attenuate animals toxins and venoms for immunizing antisera producing animals. However, the radiation effects on antigenicity and immunogenecity have not yet been elucidated. In the present work, we investigated the effects of gamma rays on the antigenic and immunogenicity have not yet been elucidated. In the present work, we investigated the effects of gamma rays on the antigenic and immunogenic behaviour of Bothrops jararacussu venon. Venom samples (2mg/ml in 150 mM NaCl) were irradiated with 500, 1000 and 2000 Gy of 60 Co gamma rays. These samples were submitted to antigen capture ELISA on plates coated with commercial bothropic antiserum. Results suggest a loss of reactivity of the 1000 and 2000 Gy irradiated samples. Antibodies against native and 2000 Gy irradiated venoms were produced in rabbits. Both sera able to bind native venom with a slightly higher titer for anti-irradiated serum. These data suggest that radiation promoted structural modification on the antigen molecules. However since the antibodies produced against irradiated antivenom were able to recognize native venom, there must have been preservation of some antigenic determinants. It has already been demosntrated that irradiation of proteins leads to structural modifications and unfolding of the molecules. Our data suggest that irradiation led to conformational epitopes destruction with preservation of linear epitopes and that the response against irradiated venom may be attributed to these linear antigenic determinants. (author). 8 refs., 3 figs

  8. Local inflammation, lethality and cytokine release in mice injected with Bothrops atrox venom

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    S. F. Barros

    1998-01-01

    Full Text Available We have provided evidence that: (a lethality of mice to crude Bothrops venom varies according the isogenic strain (A/J > C57Bl/6 > A/Sn > BALB/c > C3H/ HePas > DBA/2 > C3H/He; (bBALB/c mice (LD50=100.0 μg were injected i.p. with 50 μg of venom produced IL-6, IL-10, INF-γ, TNF-α and NO in the serum. In vitro the cells from the mice injected and challenged with the venom only released IL-10 while peritoneal macrophages released IL-10, INF-γ and less amounts of IL-6; (c establishment of local inflammation and necrosis induced by the venom, coincides with the peaks of TNF-α, IFN-γ and NO and the damage was neutralized when the venom was incubated with a monoclonal antibody against a 60 kDa haemorrhagic factor. These results suggest that susceptibility to Bothrops a trox venom is genetically dependent but MHC independent; that IL-6, IL10, TNF-α, IFN-γ and NO can be involved in the mediation of tissue damage; and that the major venom component inducers of the lesions are haemorrhagins.

  9. Screening of Bothrops snake venoms for L-amino acid oxidase activity

    Energy Technology Data Exchange (ETDEWEB)

    Pessati, M.L.; Fontana, J.D.; Guimaraes, M.F. [Federal Univ. of Parana, Curitiba (Brazil)

    1995-12-31

    Toxins, enzymes, and biologically active peptides are the main components of snake venoms from the genus Bothrops. Following the venom inoculation, the local effects are hemorrhage, edema, and myonecrosis. Nineteen different species of Brazilian Bothrops were screened for protein content and L-amino acid oxidase activity. B. cotiara, formerly found in the South of Brazil, is now threatened with extinction. Its venom contains a highly hemorrhagic fraction and, as expected from the deep yellow color of the corresponding lyophilized powder, a high L-amino acid oxidase (LAO) activity was also characterized. Flavin adenine dinucleotide (FAD) is its associate coenzyme. B. cotiara venom LAO catalyzed the oxidative deamination of several L-amino acids, and the best substrates were methionine, leucine, tryptophan, and phenylalanine, hence, its potential application for the use in biosensors for aspartame determination and for the removal of amino acids from plasma. High levels for LAO were also found in other species than B. cotiara. In addition, the technique of isoelectric focusing (IEF) was employed as a powerful tool to study the iso- or multi-enzyme distribution for LAO activity in the B. cotiara snake venom.

  10. In vitro hemolytic activity of Bothrops lanceolatus (fer-de-lance venom

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    LJ Martins

    2009-01-01

    Full Text Available Bothrops lanceolatus venom contains a variety of enzymatic and biological activities. The present work investigated the hemolytic activity of this venom and its phospholipase A2 (PLA2. Bothrops lanceolatus venom (6.7 µg/mL caused indirect hemolysis of cow, horse, rat and sheep erythrocytes, with horse erythrocytes being the most sensitive; no direct hemolysis was observed. Hemolysis in sheep erythrocytes was concentration-dependent (5-11.7 µg/mL and markedly attenuated by heating the venom for 30 minutes at ≥ 40°C and by the PLA2 inhibitor p-bromophenacyl bromide. An acidic PLA2 (5 µg/mL purified from B. lanceolatus venom also caused hemolysis. This PLA2 showed immunoprecipitin lines with antivenom against B. lanceolatus, which suggests that the enzymatic and hemolytic activities of this enzyme may be neutralized during antivenom therapy. These results indicate that B. lanceolatus venom and its PLA2 can cause hemolysis in vitro.

  11. The Triterpenoid Betulin Protects against the Neuromuscular Effects of Bothrops jararacussu Snake Venom In Vivo

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    Miriéle Cristina Ferraz

    2015-01-01

    Full Text Available We confirmed the ability of the triterpenoid betulin to protect against neurotoxicity caused by Bothrops jararacussu snake venom in vitro in mouse isolated phrenic nerve-diaphragm (PND preparations and examined its capability of in vivo protection using the rat external popliteal/sciatic nerve-tibialis anterior (EPSTA preparation. Venom caused complete, irreversible blockade in PND (40 μg/mL, but only partial blockade (~30% in EPSTA (3.6 mg/kg, i.m. after 120 min. In PND, preincubation of venom with commercial bothropic antivenom (CBA attenuated the venom-induced blockade, and, in EPSTA, CBA given i.v. 15 min after venom also attenuated the blockade (by ~70% in both preparations. Preincubation of venom with betulin (200 μg/mL markedly attenuated the venom-induced blockade in PND; similarly, a single dose of betulin (20 mg, i.p., 15 min after venom virtually abolished the venom-induced decrease in contractility. Plasma creatine kinase activity was significantly elevated 120 min after venom injection in the EPSTA but was attenuated by CBA and betulin. These results indicate that betulin given i.p. has a similar efficacy as CBA given i.v. in attenuating the neuromuscular effects of B. jararacussu venom in vivo and could be a useful complementary measure to antivenom therapy for treating snakebite.

  12. The Triterpenoid Betulin Protects against the Neuromuscular Effects of Bothrops jararacussu Snake Venom In Vivo.

    Science.gov (United States)

    Ferraz, Miriéle Cristina; de Oliveira, Jhones Luiz; de Oliveira Junior, Joel Reis; Cogo, José Carlos; Dos Santos, Márcio Galdino; Franco, Luiz Madaleno; Puebla, Pilar; Ferraz, Helena Onishi; Ferraz, Humberto Gomes; da Rocha, Marisa Maria Teixeira; Hyslop, Stephen; San Feliciano, Arturo; Oshima-Franco, Yoko

    2015-01-01

    We confirmed the ability of the triterpenoid betulin to protect against neurotoxicity caused by Bothrops jararacussu snake venom in vitro in mouse isolated phrenic nerve-diaphragm (PND) preparations and examined its capability of in vivo protection using the rat external popliteal/sciatic nerve-tibialis anterior (EPSTA) preparation. Venom caused complete, irreversible blockade in PND (40 μg/mL), but only partial blockade (~30%) in EPSTA (3.6 mg/kg, i.m.) after 120 min. In PND, preincubation of venom with commercial bothropic antivenom (CBA) attenuated the venom-induced blockade, and, in EPSTA, CBA given i.v. 15 min after venom also attenuated the blockade (by ~70% in both preparations). Preincubation of venom with betulin (200 μg/mL) markedly attenuated the venom-induced blockade in PND; similarly, a single dose of betulin (20 mg, i.p., 15 min after venom) virtually abolished the venom-induced decrease in contractility. Plasma creatine kinase activity was significantly elevated 120 min after venom injection in the EPSTA but was attenuated by CBA and betulin. These results indicate that betulin given i.p. has a similar efficacy as CBA given i.v. in attenuating the neuromuscular effects of B. jararacussu venom in vivo and could be a useful complementary measure to antivenom therapy for treating snakebite. PMID:26633987

  13. Paper electrophoretic and enzimatic studies on blood serum, venom and liver of "Bothrops jararaca"

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    Luiz P. Ribeiro

    1955-12-01

    Full Text Available Fígado, veneno e sôro sanguíneo de "Bothrops jararaca" foram estudados por meio da eletroforese em papel e determinação de atividades enzimáticas. Xantina oxidase e deshidrogenase foram encontradas sòmente no fígado das cobras. A análise espectrográfica do veneno e do sôro confirmam os resultados negativos obtidos para xantina oxidase uma vez que não foi encontrado molibdêneo. L-amino ácido oxidase foi determinada no fígado, sÔro e veneno. A eletroforese em papel do sôro sanguíneo mostrou que existem 7 frações proteicas, sendo que duas apresentam fluorescência característica de flavinas, quando expostas à luz ultra-violeta. Em vista dos resultados obtidos é concluido que as flavinas do sôro e do veneno de Bothrops jararaca estão na maior parte ligadas às proteínas. Estas flavinas combinadas parecem estar sob a forma de FAD (flavina adenina dinucleotídeo fazendo parte do grupo prostético da L-amino ácido oxidase, uma vez que não foi encontrada nenhuma atividade de xantina oxidase.

  14. Activation of J77A.1 Macrophages by Three Phospholipases A2 Isolated from Bothrops atrox Snake Venom

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    Juliana L. Furtado

    2014-01-01

    Full Text Available In the present study, we investigated the in vitro effects of two basic myotoxic phospholipases A2 (PLA2, BaTX-I, a catalytically inactive Lys-49 variant, and BaTX-II, a catalytically active Asp-49, and of one acidic myotoxic PLA2, BaPLA2, a catalytically active Asp-49, isolated from Bothrops atrox snake venom, on the activation of J774A.1 macrophages. At noncytotoxic concentrations, the toxins did not affect the adhesion of the macrophages, nor their ability to detach. The data obtained showed that only BaTX-I stimulated complement receptor-mediated phagocytosis. However, BaTX-I, BaTX-II, and BaPLA2 induced the release of the superoxide anion by J774A.1 macrophages. Additionally, only BaTX-I raised the lysosomal volume of macrophages after 15 min of incubation. After 30 min, all the phospholipases increased this parameter, which was not observed within 60 min. Moreover, BaTX-I, BaTX-II, and BaPLA2 increased the number of lipid bodies on macrophages submitted to phagocytosis and not submitted to phagocytosis. However, BaTX-II and BaPLA2 induced the release of TNF-α by J774A.1 macrophages. Taken together, the data show that, despite differences in enzymatic activity, the three toxins induced inflammatory events and whether the enzyme is acidic or basic does not seem to contribute to these effects.

  15. Activation of J77A.1 macrophages by three phospholipases A2 isolated from Bothrops atrox snake venom.

    Science.gov (United States)

    Furtado, Juliana L; Oliveira, George A; Pontes, Adriana S; Setúbal, Sulamita da S; Xavier, Caroline V; Lacouth-Silva, Fabianne; Lima, Beatriz F; Zaqueo, Kayena D; Kayano, Anderson M; Calderon, Leonardo A; Stábeli, Rodrigo G; Soares, Andreimar M; Zuliani, Juliana P

    2014-01-01

    In the present study, we investigated the in vitro effects of two basic myotoxic phospholipases A2 (PLA2), BaTX-I, a catalytically inactive Lys-49 variant, and BaTX-II, a catalytically active Asp-49, and of one acidic myotoxic PLA2, BaPLA2, a catalytically active Asp-49, isolated from Bothrops atrox snake venom, on the activation of J774A.1 macrophages. At noncytotoxic concentrations, the toxins did not affect the adhesion of the macrophages, nor their ability to detach. The data obtained showed that only BaTX-I stimulated complement receptor-mediated phagocytosis. However, BaTX-I, BaTX-II, and BaPLA2 induced the release of the superoxide anion by J774A.1 macrophages. Additionally, only BaTX-I raised the lysosomal volume of macrophages after 15 min of incubation. After 30 min, all the phospholipases increased this parameter, which was not observed within 60 min. Moreover, BaTX-I, BaTX-II, and BaPLA2 increased the number of lipid bodies on macrophages submitted to phagocytosis and not submitted to phagocytosis. However, BaTX-II and BaPLA2 induced the release of TNF-α by J774A.1 macrophages. Taken together, the data show that, despite differences in enzymatic activity, the three toxins induced inflammatory events and whether the enzyme is acidic or basic does not seem to contribute to these effects. PMID:24592395

  16. Preclinical assessment of the neutralizing capacity of antivenoms produced in six Latin American countries against medically-relevant Bothrops snake venoms.

    Science.gov (United States)

    Segura, A; Castillo, M C; Núñez, V; Yarlequé, A; Gonçalves, L R C; Villalta, M; Bonilla, C; Herrera, M; Vargas, M; Fernández, M; Yano, M Y; Araújo, H P; Boller, M A A; León, P; Tintaya, B; Sano-Martins, I S; Gómez, A; Fernández, G P; Geoghegan, P; Higashi, H G; León, G; Gutiérrez, J M

    2010-11-01

    Species of the genus Bothrops induce the vast majority of snakebite envenomings in Latin America. A preclinical study was performed in the context of a regional network of public laboratories involved in the production, quality control and development of antivenoms in Latin America. The ability of seven polyspecific antivenoms, produced in Argentina, Brazil, Peru, Bolivia, Colombia and Costa Rica, to neutralize lethal, hemorrhagic, coagulant, defibrinogenating and myotoxic activities of the venoms of Bothrops neuwiedi (diporus) (Argentina), Bothrops jararaca (Brazil), B. neuwiedi (mattogrossensis) (Bolivia), Bothrops atrox (Peru and Colombia) and Bothrops asper (Costa Rica) was assessed using standard laboratory tests. Despite differences in the venom mixtures used in the immunization of animals for the production of these antivenoms, a pattern of extensive cross-neutralization was observed between these antivenoms and all the venoms tested, with quantitative differences in the values of effective doses. This study reveals the capacity of these antivenoms to neutralize, in preclinical tests, homologous and heterologous Bothrops venoms in Central and South America, and also highlight quantitative differences in the values of Median Effective Doses (ED50s) between the various antivenoms.

  17. Preclinical assessment of the neutralizing capacity of antivenoms produced in six Latin American countries against medically-relevant Bothrops snake venoms.

    Science.gov (United States)

    Segura, A; Castillo, M C; Núñez, V; Yarlequé, A; Gonçalves, L R C; Villalta, M; Bonilla, C; Herrera, M; Vargas, M; Fernández, M; Yano, M Y; Araújo, H P; Boller, M A A; León, P; Tintaya, B; Sano-Martins, I S; Gómez, A; Fernández, G P; Geoghegan, P; Higashi, H G; León, G; Gutiérrez, J M

    2010-11-01

    Species of the genus Bothrops induce the vast majority of snakebite envenomings in Latin America. A preclinical study was performed in the context of a regional network of public laboratories involved in the production, quality control and development of antivenoms in Latin America. The ability of seven polyspecific antivenoms, produced in Argentina, Brazil, Peru, Bolivia, Colombia and Costa Rica, to neutralize lethal, hemorrhagic, coagulant, defibrinogenating and myotoxic activities of the venoms of Bothrops neuwiedi (diporus) (Argentina), Bothrops jararaca (Brazil), B. neuwiedi (mattogrossensis) (Bolivia), Bothrops atrox (Peru and Colombia) and Bothrops asper (Costa Rica) was assessed using standard laboratory tests. Despite differences in the venom mixtures used in the immunization of animals for the production of these antivenoms, a pattern of extensive cross-neutralization was observed between these antivenoms and all the venoms tested, with quantitative differences in the values of effective doses. This study reveals the capacity of these antivenoms to neutralize, in preclinical tests, homologous and heterologous Bothrops venoms in Central and South America, and also highlight quantitative differences in the values of Median Effective Doses (ED50s) between the various antivenoms. PMID:20621114

  18. N-terminal domain of Bothrops asper Myotoxin II Enhances the Activity of Endothelin Converting Enzyme-1 and Neprilysin

    Science.gov (United States)

    Smith, A. Ian; Rajapakse, Niwanthi W.; Kleifeld, Oded; Lomonte, Bruno; Sikanyika, Nkumbu L.; Spicer, Alexander J.; Hodgson, Wayne C.; Conroy, Paul J.; Small, David H.; Kaye, David M.; Parkington, Helena C.; Whisstock, James C.; Kuruppu, Sanjaya

    2016-01-01

    Neprilysin (NEP) and endothelin converting enzyme-1 (ECE-1) are two enzymes that degrade amyloid beta in the brain. Currently there are no molecules to stimulate the activity of these enzymes. Here we report, the discovery and characterisation of a peptide referred to as K49-P1-20, from the venom of Bothrops asper which directly enhances the activity of both ECE-1 and NEP. This is evidenced by a 2- and 5-fold increase in the Vmax of ECE-1 and NEP respectively. The K49-P1-20 concentration required to achieve 50% of maximal stimulation (AC50) of ECE-1 and NEP was 1.92 ± 0.07 and 1.33 ± 0.12 μM respectively. Using BLITZ biolayer interferometry we have shown that K49-P1-20 interacts directly with each enzyme. Intrinsic fluorescence of the enzymes change in the presence of K49-P1-20 suggesting a change in conformation. ECE-1 mediated reduction in the level of endogenous soluble amyloid beta 42 in cerebrospinal fluid is significantly higher in the presence of K49-P1-20 (31 ± 4% of initial) compared with enzyme alone (11 ± 5% of initial; N = 8, P = 0.005, unpaired t-test). K49-P1-20 could be an excellent research tool to study mechanism(s) of enzyme stimulation, and a potential novel drug lead in the fight against Alzheimer’s disease. PMID:26931059

  19. Ontogenetic variation of metalloproteinases and plasma coagulant activity in venoms of wild Bothrops atrox specimens from Amazonian rain forest.

    Science.gov (United States)

    López-Lozano, Jorge Luis; de Sousa, Marcelo Valle; Ricart, Carlos André O; Chávez-Olortegui, Carlos; Flores Sanchez, Eladio; Muniz, Emiro G; Bührnheim, Paulo F; Morhy, Lauro

    2002-07-01

    A comparative study of venoms from juvenile, sub-adult and adult wild Bothrops atrox specimens captured in Manaus region (Brazil) was performed. All venoms tested had acidic pH (5.5) and the human plasma coagulant activity was higher in venoms from juvenile and sub-adult specimens than in adults. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the most intense bands in adult venoms corresponded to polypeptides of 23 and 50kDa. The 23kDa protein was not detected in juvenile venoms. The 23 and 50kDa proteins were purified by two steps of reversed phase-HPLC followed by size exclusion HPLC. Partial amino acid sequence of the 23kDa protein showed homology to metalloproteinases from other snake venoms. Electrospray ionization mass spectrometric analysis (ESI-MS) showed that the 23kDa band contained at least three isoforms of 23030, 23300 and 23645Da. The 50kDa polypeptide was N-terminally blocked for Edman degradation and presented molecular masses ranging from 46.8 to 49.4kDa by ESI-MS. Both proteins were detected by anti-mutalysin II antibodies in immunoblotting assay indicating that they belong to the metalloproteinase family. Immunoblotting analysis also showed that the 23kDa band increased in intensity from juvenile to adult specimens.SDS-PAGE analysis of juvenile and adult venoms following autoproteolysis in pH 7.4 suggested that endogenous venom metalloproteinases can digest the 50kDa metalloproteinase, originating a new protein band of 27kDa. It was also demonstrated in juvenile venoms that the 23kDa band was not the result of proteolytic processing of the 50kDa metalloproteinase. PMID:12076654

  20. Variaciones en las actividades enzimáticas del veneno de la serpiente Bothrops atrox "jergón", de tres zonas geográficas del Perú Variation of the enzymatic activity of Bothrops atrox "jergon" snake venom from three geographic regions, Peru

    Directory of Open Access Journals (Sweden)

    César Ortiz

    2012-06-01

    casein and by zymogram. Additionally, immunodiffusion and neutralization assays in vitro were done with a polyvalent botropic serum from the national institute of health of Peru. Results. The amidolytic, coagulant, hemorrhagic, proteolytic by zymogram, phospholipase A2, and indirect hemolytic activity were variable, demonstrating increased activity in the venoms from Amazonas, regarding proteolytic by zymogram, phospholipase A2, and indirect hemolytic activity. While the amount of protein electrophoretic bands and proteolytic activity on casein did not demonstrated differences. Regarding neutralization tests, a 0.5 dose of antivenom was sufficient to effectively neutralize (>50% the coagulant activity and phospholipase A2 of all samples analyzed. Conclusions. Some biological properties of the venom from adult Bothrops atrox of Peru are variable, without interference with the in vitro neutralization by the polyvalent botropic serum on coagulant and phospholipase A2 properties of the venom.

  1. Visual cues for the host-finding and mating locations of Monochamus alternatus ( Coleoptera : Cerambycidae)%视觉因素在松墨天牛定位寄主和交配中的作用

    Institute of Scientific and Technical Information of China (English)

    刘博; 徐华潮; 孟俊国; 孙江华; 樊建庭

    2012-01-01

    In order to explore the role of the visual factors, for Monochamus alternatus (Coleoptera: Ceramby cidae) four treatments of field traps (black, brown, green, and gray-control) were tested. The mating behavior was observed after M. alternatus' visual organ compound eyes and olfaction were treated. Results showed that the black and brown traps were stronger than the gray (control) traps (P<0.05); but the green traps were weaker than the control (P<0.05=. In mating behavior assays, after the compound eyes were treated, the percentage of mating success decreased to 50.7% compared to the control which had an average of 93.3%. Also, the proportion of mating success after treating olfaction (antenna were cutting down) averaged 72.3%, which was lower than the control (P<0.05=. With both vision and olfaction treated, the proportion of mating success averaged 26.7%, also lower than the control (P<0.05). This indicated that visual cues over short distances were very important in the mating behavior of M. alternatus. [Ch, 2 fig. 24 ref. ]%在野外诱捕试验中,使用相同的诱芯,测试了4种不同颜色诱捕器对松墨天牛Monochamus alternatus的引诱效果.其中黑色和棕色诱捕器引诱效果显著高于对照灰白色,而绿色诱捕器引诱效果最差,显著低于对照.这表明,在松墨天牛的野外远距离寄主定位过程中,视觉因素起到了重要的作用,黑色和棕色提高了引诱效果,而绿色抑制了引诱效果.在室内交配定位试验中,当松墨天牛的复眼被涂黑处理之后,雌雄抱对交配的成功率降低为50.7%,显著低于对照健康组的成功率(933%).当松墨天牛触角被切除之后,抱对交配成功率为72.3%,同样显著低于对照.而将天牛的复眼涂黑并且切除触角之后,抱对交配的成功率仅为26.7%.视觉因素在天牛近距离交配定位过程中发挥着重要的作用.

  2. Ocorrência de híbridos não naturais entre Bothrops jararaca e B. neuwiedi (Serpentes, Viperidae Occurrence of non natural hybridism between Bothrops jararaca and B. neuwiedi (Serpentes, Viperidae

    Directory of Open Access Journals (Sweden)

    Rafael Lucchesi Balestrin

    Full Text Available In May 1987, a female of Bothrops jararaca (Wied, 1824, from Carazinho, Rio Grande do Sul (RS, Brazil, was placed in the same vivarium with a male of Bothrops neuwiedi Wagler, 1824 coming from Guaíba, RS. There, they stayed for aproximately ten months. In March 1988, it was observed a delivery of five live and two still born, among them six presented morphologic characteristics of B. neuwiedi and one of B. jararaca. After the female died, in April 1988, through necropsy, two fetusus were found, one near the cloaca and, both identified as B. neuwiedi. The morphologic analysis and the origin of the progenitors suggest the hypothesis that the litter was resulted of cross-breeding.

  3. 松墨天牛成虫标本保存及其DNA提取质量比较%Comparison of the quality of genomic DNA extracted from adult specimens of Monochamus alternatus preserved by different methods

    Institute of Scientific and Technical Information of China (English)

    曲良建; 王丽娟; 王青华; 王玉珠; 张永安

    2014-01-01

    Objectives] To find which method of preserving Monochamus alternatus specimens produced the best quality genomic DNA. [Methods] Genomic DNA from Monochamus alternatus adults that had been preserved using different methods (liquid nitrogen, 100%ethanol at-20℃, 100%ethanol at room temperature and museum specimens) for more than 2 years, was extracted using the SDS-proteinase K method and the quality of the extracted genomic DNA compared. [Results] The best quality genomic DNA was obtained from specimens that had been stored in liquid nitrogen, and the next best from those that had been preserved in 100%ethanol at-20℃. [Conclusion] The best quality DNA was extracted from specimens that had been frozen in liquid nitrogen or preserved in 100%ethanol at-20℃. The DNA samples obtained from such specimens are suitable for PCR amplification and sequencing.%【目的】探讨适合DNA 提取的天牛成虫标本保存方法。【方法】采用SDS-蛋白酶K消化法对液氮中冷冻保存、无水乙醇-20℃冷冻保存、无水乙醇室温保存和干标本室温保存且保存时间在2年以上的松墨天牛Monochamus alternates Hope成虫标本基因组DNA进行提取,并对不同保存方式提取的DNA样本进行了质量比较和分析。【结果】在上述常见的松墨天牛成虫标本4种保存方式中,以液氮中冷冻保存效果最佳,其次为无水乙醇-20℃冷冻保存,插针干标本室温保藏效果最差。利用昆虫线粒体基因COⅠ和COⅡ的通用引物从上述DNA中均能够成功扩增出目的片段,测序结果证实扩增片段符合预期。【结论】液氮和无水乙醇-20℃冷冻保存适合松墨天牛成虫标本长期保存,且不影响后续的PCR扩增和测序。

  4. Purificación de fosfolipas A2 a partir de veneno de Bothrops atrox

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    Quevedo B.

    1999-06-01

    Full Text Available La fosfolipasa A2 (PLA2 fue purificada a partir de veneno de serpiente de Bothrops atrox (Sensu lato de Chiriguaná (Colombia por cromatografía de exclusión sobre Sephadex G-75, de donde se obtuvieron cinco fracciones, una de ellas con actividad fosfolipasa A2. Después de pasarla por una columna de intercambio catiónico Mono S, donde se separaron ocho fracciones con activi-dad PLA2, ésta se midió usando el método hemolítico. La electroforesis en gel de poliacrilamida en condiciones desnaturalizantes, mostró pesos moleculares entre 16.000 y 17.000 para las ocho fracciones, seis de las cuales pre-sentaron una sola banda. 

  5. Cross-reactivity and phospholipase A2 neutralization of anti-irradiated Bothrops jararaca venom antibodies

    International Nuclear Information System (INIS)

    The detoxified Bothrops jararaca venom, immunized rabbits with the toxoid obtained and investigated cross-reactivity of the antibodies obtained against autologous and heterelogous venoms was presented. It was also investigated the ability of the IgGs, purified by affinity chromatography, from those sera to neutralize phospholipase. A2, an ubiquous enzyme in animal venoms. Results indicate that venom irradiation leads to an attenuation of toxicity of 84%. Cross-reactivity was investigated by ELISA and Western blot and all venoms were reactive to the antibodies. On what refers to phospholipase A2 activity neutralization, the antibodies neutralized autologous venoms efficiently and, curiously, other venoms from the same genus were not neutralized, while Lachesis muta venom, a remote related specier, was neutralized by this serum. These data suggest that irradiation preserve important epitopes for induction of neutralizing antibodies and that these epitopes are not shared by all venoms assayed. (author). 8 refs, 2 figs, 3 tabs

  6. Neutralizing capacity of a new monovalent anti-Bothrops atrox antivenom: comparison with two commercial antivenoms

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    R. Otero

    1997-03-01

    Full Text Available Three horse-derived antivenoms were tested for their ability to neutralize lethal, hemorrhagic, edema-forming, defibrinating and myotoxic activities induced by the venom of Bothrops atrox from Antioquia and Chocó (Colombia. The following antivenoms were used: a polyvalent (crotaline antivenom produced by Instituto Clodomiro Picado (Costa Rica, b monovalent antibothropic antivenom produced by Instituto Nacional de Salud-INS (Bogotá, and c a new monovalent anti-B. atrox antivenom produced with the venom of B. atrox from Antioquia and Chocó. The three antivenoms neutralized all toxic activities tested albeit with different potencies. The new monovalent anti-B. atrox antivenom showed the highest neutralizing ability against edema-forming and defibrinating effects of B. atrox venom (41 ± 2 and 100 ± 32 µl antivenom/mg venom, respectively, suggesting that it should be useful in the treatment of B. atrox envenomation in Antioquia and Chocó

  7. Local inflammation, lethality and cytokine release in mice injected with Bothrops atrox venom.

    OpenAIRE

    Barros, S F; I. Friedlanskaia; V. L. Petricevich; Kipnis, T L

    1998-01-01

    We have provided evidence that: (a) lethality of mice to crude Bothrops venom varies according the isogenic strain (A/J > C57Bl/6 > A/Sn > BALB/c > C3H/HePas > DBA/2 > C3H/He); (b)BALB/c mice (LD50=100.0 microg) were injected i.p. with 50 microg of venom produced IL-6, IL-10, INF-gamma, TNF-alpha and NO in the serum. In vitro the cells from the mice injected and challenged with the venom only released IL-10 while peritoneal macrophages released IL-10, INF-gamma and less amounts of IL-6; (c) e...

  8. Thrombocytin, a serine protease from Bothrops atrox venom. 1. Purification and characterization of the enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Kirby, E.P. (Temple Univ. Health Sciences Center, Philadelphia, PA); Niewiarowski, S.; Stocker, K.; Kettner, C.; Shaw, E.; Brudzynsi, T.M.

    1979-08-07

    Thrombocytin, a platelet-activating enzyme from Bothrops atrox venom, has been purified to homogeneity by precipitation with sodium salicylate and chromatography on heparin-agarose. Thrombocytin is a single-chain glycoprotein with a molecular weight of 36,000 which contains 5.6% carbohydrate. It causes platelet aggregation, release of platelet serotonin, and activation of factor XIII. The most sensitive substrate for the amidolytic activity of thrombocytin was Tos-Gly-Pro-Arg-p-nitroanilide hydrochloride. The activity of thrombocytin on this substrate and on platelets was inhibited by diisopropyl fluorophosphate (DFP), soybean trypsin inhibitor, and several arginine chloromethyl ketones. Active site titration with nitrophenyl guanidinobenzoate demonstrated that approximately 86% of the preparation was in the active form. These experiments demonstrate the presence of serine and histidine in the active site of thrombocytin and suggest that thrombocytin is a classical serine protease with a platelet-activating activity similar to thrombin.

  9. Local and systemic biochemical alterations induced by Bothrops atrox snake venom in mice.

    Science.gov (United States)

    de Souza, Carlos At; Kayano, Anderson M; Setúbal, Sulamita S; Pontes, Adriana S; Furtado, Juliana L; Kwasniewski, Fábio H; Zaqueo, Kayena D; Soares, Andreimar M; Stábeli, Rodrigo G; Zuliani, Juliana P

    2012-01-01

    The local and systemic alterations induced by Bothrops atrox snake venom (BaV) injection in mice were studied. BaV induced superoxide production by migrated neutrophils, mast cell degranulation and phagocytosis by macrophages. Moreover, BaV caused hemorrhage in dorsum of mice after 2hr post- injection. Three hours post-injection in gastrocnemius muscle, we also observed myonecrosis, which was assessed by the determination of serum and tissue CK besides the release of urea, but not creatinine and uric acid, indicating kidney alterations. BaV also induced the release of LDH and transaminases (ALT and AST) indicating tissue and liver abnormalities. In conclusion, the data indicate that BaV induces events of local and systemic importance. PMID:23487552

  10. Glial cells of the central nervous system of Bothrops jararaca (Reptilia, Ofidae: an ultrastructural study

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    Eduardo F. Bondan

    2015-07-01

    Full Text Available Abstract Although ultrastructural characteristics of mature neuroglia in the central nervous system (CNS are very well described in mammals, much less is known in reptiles, especially serpents. In this context, two specimens of Bothrops jararaca were euthanized for morphological analysis of CNS glial cells. Samples from telencephalon, mesencephalon and spinal cord were collected and processed for light and transmission electron microscopy investigation. Astrocytes, oligodendrocytes, microglial cells and ependymal cells, as well as myelin sheaths, presented similar ultrastructural features to those already observed in mammals and tended to maintain their general aspect all over the distinct CNS regions observed. Morphological similarities between reptilian and mammalian glia are probably linked to their evolutionary conservation throughout vertebrate phylogeny.

  11. Phylogenetic analysis of β-defensin-like genes of Bothrops, Crotalus and Lachesis snakes.

    Science.gov (United States)

    Correa, Poliana G; Oguiura, Nancy

    2013-07-01

    Defensins are components of the vertebrate innate immune system; they comprise a diverse group of small cationic antimicrobial peptides. Among them, β-defensins have a characteristic β-sheet-rich fold plus six conserved cysteines with particular spacing and intramolecular bonds. They have been fully studied in mammals, but there is little information about them in snakes. Using a PCR approach, we described 13 β-defensin-like sequences in Bothrops and Lachesis snakes. The genes are organized in three exons and two introns, with exception of B.atrox_defensinB_01 which has only two exons. They show high similarities in exon 1, intron 1 and intron 2, but exons 2 and 3 have undergone accelerated evolution. The theoretical translated sequences encode a pre-β-defensin-like molecule with a conserved signal peptide and a mature peptide. The signal peptides are leucine-rich and the mature β-defensin-like molecules have a size around 4.5 kDa, a net charge from +2 to +11, and the conserved cysteine motif. Phylogenetic analysis was done using maximum parsimony, maximum likelihood and Bayesian analyses, and all resulted in similar topologies with slight differences. The genus Bothrops displayed two separate lineages. The reconciliation of gene trees and species tree indicated eight to nine duplications and 23 to 29 extinctions depending on the gene tree used. Our results together with previously published data indicate that the ancestral β-defensin-like gene may have three exons in vertebrates and that their evolution occurred according to a birth-and-death model. PMID:23500066

  12. Comparative evaluation of adverse effects in the use of powder trivalent antivenom and liquid antivenoms in Bothrops snake bites

    OpenAIRE

    Iran Mendonça da Silva; Antônio Magela Tavares

    2012-01-01

    INTRODUCTION: Snake bite, a problem in public health, generally occurs where there is no electric power. METHODS: A comparative clinical study was conducted with 102 victims of Bothrops snake bite, from the state of Amazonas, Brazil; 58 victims were treated with liofilizated trivalent antivenom serum (SATL) and 44 victims treated with liquid bivalent and monovalent antivenom serum (SAMBL). RESULTS: 17% (10/58) of patients presented adverse effects with the SATL and 25% (11/44) with the SAMBL....

  13. Occurrence of sulfated fucose branches in fucosylated chondroitin sulfate are essential for the polysaccharide effect preventing muscle damage induced by toxins and crude venom from Bothrops jararacussu snake.

    Science.gov (United States)

    Monteiro-Machado, Marcos; Tomaz, Marcelo A; Fonseca, Roberto J C; Strauch, Marcelo A; Cons, Bruno L; Borges, Paula A; Patrão-Neto, Fernando C; Tavares-Henriques, Matheus S; Teixeira-Cruz, Jhonatha M; Calil-Elias, Sabrina; Cintra, Adélia C O; Martinez, Ana Maria B; Mourão, Paulo A S; Melo, Paulo A

    2015-05-01

    Snake envenoming is an important public health problem around the world, particularly in tropics. Beyond deaths, morbidity induced by snake venoms, such as myotoxicity, is of pivotal consequence to population. Bothrops jararacussu is the main venomous snake in southeast region of Brazil, and particularly presents strong myotoxic effect. The only available therapy, antibothropic antivenom, poorly affects venom-induced myotoxicity. The aim of this study is to assess the ability of fucosylated chondroitin sulfate (fucCS), a glycosaminoglycan with anticoagulant and antithrombotic properties, and its derivatives to inhibit toxic activities of B. jararacussu crude venom and its isolated toxins, named bothropstoxins (BthTX-I and BthTX-II). The in vitro myotoxic activities induced by crude venom, by BthTX-I alone and by toxins together were abolished by fucCS. Carboxyl reduction (fucCS-CR) kept this ability whereas defucosilation (defucCS) abrogates myoprotection. We observed the same pattern in the response of these polysaccharides in antagonizing the increase in plasma creatine kinase (CK) levels, the reduction of skeletal muscle CK content and the rise of myeloperoxidase (MPO) activity induced by crude venom and isolated toxins. FucCS inhibited edematogenic activity and partially prevented the reduction of total leukocytes in blood when pre-incubated with crude venom. Furthermore, the venom procoagulant effect was completely antagonized by increasing concentrations of fucCS, although this polyanion could stop neither the tail bleeding nor the skin hemorrhage induced by Bothrops jararaca venom. The B. jararacussu phospholipase, hyaluronidase, proteolytic and collagenase activities were inhibited in vitro. The results suggest that fucCS could be able to interact with both toxins, and it is able to inhibit BthTX-II phospholipase activity. Light microscopy of extensor digitorum longus muscle (EDL) muscle showed myoprotection by fucCS, once necrotic areas, edema and

  14. Phylogeography of the Bothrops jararaca complex (Serpentes: Viperidae): past fragmentation and island colonization in the Brazilian Atlantic Forest.

    Science.gov (United States)

    Grazziotin, Felipe G; Monzel, Markus; Echeverrigaray, Sergio; Bonatto, Sandro L

    2006-11-01

    The Brazilian Atlantic Forest is one of the world's major biodiversity hotspots and is threatened by a severe habitat loss. Yet little is known about the processes that originated its remarkable richness of endemic species. Here we present results of a large-scale survey of the genetic variation at the mitochondrial cytochrome b gene of the pitviper, jararaca lancehead (Bothrops jararaca), and two closely related insular species (Bothrops insularis and Bothrops alcatraz), endemic of this region. Phylogenetic and network analyses revealed the existence of two well-supported clades, exhibiting a southern and a northern distribution. The divergence time of these two phylogroups was estimated at 3.8 million years ago, in the Pliocene, a period of intense climatic changes and frequent fragmentation of the tropical rainforest. Our data also suggest that the two groups underwent a large size expansion between 50,000 and 100,000 years ago. However, the southern group showed a more marked signal of population size fluctuation than the northern group, corroborating evidences that southern forests may have suffered a more pronounced reduction in area in the late Pleistocene. The insular species B. alcatraz and B. insularis presented very low diversity, each one sharing haplotypes with mainland individuals placed in different subclades. Despite their marked morphological and behavioural uniqueness, these two insular species seem to have originated very recently and most likely from distinct costal B. jararaca populations, possibly associated with late Pleistocene or Holocene sea level fluctuations. PMID:17054497

  15. Low-level laser therapy decreases local effects induced by myotoxins isolated from Bothrops jararacussu snake venom

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    AM Barbosa

    2010-01-01

    Full Text Available The prominent myotoxic effects induced by Bothrops jararacussu crude venom are due, in part, to its polycationic myotoxins, BthTX-I and BthTX-II. Both myotoxins have a phospholipase A2 structure: BthTX-II is an active enzyme Asp-49 PLA2, while BthTX-I is a Lys-49 PLA2 devoid of enzymatic activity. In this study, the effect of low-level laser therapy (LLLT, 685 nm laser at a dose of 4.2 J/cm2 on edema formation, leukocyte influx and myonecrosis caused by BthTX-I and BthTX-II, isolated from Bothrops jararacussu snake venom, was analyzed. BthTX-I and BthTX-II caused a significant edema formation, a prominent leukocyte infiltrate composed predominantly by neutrophils and myonecrosis in envenomed gastrocnemius muscle. LLLT significantly reduced the edema formation, neutrophil accumulation and myonecrosis induced by both myotoxins 24 hours after the injection. LLLT reduced the myonecrosis caused by BthTX-I and BthTX-II, respectively, by 60 and 43%; the edema formation, by 41 and 60.7%; and the leukocyte influx, by 57.5 and 51.6%. In conclusion, LLLT significantly reduced the effect of these snake toxins on the inflammatory response and myonecrosis. These results suggest that LLLT should be considered a potential therapeutic approach for treatment of local effects of Bothrops species venom.

  16. ENVENENAMIENTO OFIDICO POR EL GENERO Bothrops COMPLICADO CON MIOCARDIOPATIA TÓXICA: A PROPOSITO DE UN CASO.

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    Sajar Abusaid Palomo

    2014-12-01

    Full Text Available El envenenamiento ofídico en Venezuela es un problema de salud pública, afectando a agricultores y mineros, implicada con mayor frecuencia la familia Viperidae, genero Bothrops. Presenta clínica variable, desde manifestaciones locales hasta sistémicas. Paciente femenina de 40 años, agricultora, consulta por dolor y aumento de volumen en cara lateral de pie izquierdo por envenenamiento ofídico. Acude a ambulatorio de su localidad, 24 horas después es trasladada al Hospital Universitario Ruiz y Páez, ingresando con diagnóstico de Envenenamiento Ofídico por Bothrops. Evoluciona tórpidamente presentando Insuficiencia Renal Aguda en terapia de hemodiálisis, Sepsis punto de partida respiratorio, Insuficiencia Respiratoria Aguda y Miocardiopatía Tóxica, por lo que ingresa a Unidad de Cuidados Intensivos y Cuidados Coronarios. Luego de marcada mejoría clínica es dada de alta. El envenenamiento ofídico por Bothrops corresponde 80% de los accidentes por mordeduras de serpientes, su pronóstico depende de la especie, atención medica-hospitalaria, el intervalo de tiempo ocurrido y el uso de la terapia antiofídica especifica.

  17. Prognostic significance of clinical grading of patients envenomed by Bothrops lanceolatus in Martinique. Members of the Research Group on Snake Bite in Martinique.

    Science.gov (United States)

    Thomas, L; Tyburn, B; Ketterlé, J; Biao, T; Mehdaoui, H; Moravie, V; Rouvel, C; Plumelle, Y; Bucher, B; Canonge, D; Marie-Nelly, C A; Lang, J

    1998-01-01

    The correlation between clinical grading of patients bitten by Bothrops lanceolatus and the subsequent development of their envenoming was examined. Severity of envenoming was graded using a 1-4 scale (minor to major). Patients were classified into 2 groups according to the time elapsed between bite and treatment with a specific purified equine F(ab')2 antivenom. The late/no treatment group (n = 33) was characterized by a systemic thrombotic complication rate of 14/33 (42.4%) leading to 4 deaths, which increased with the maximum severity assessed on the first day following the bite (P = 0.003). However, infarctions could develop in patients who presented initially with signs of moderate envenoming, normal blood clotting and low serum levels of venom antigens. No such complication of fatality occurred in the early (0.5-6 h) treatment group (n = 70). Multiple regression analysis showed that duration of stay in hospital in this group increased with the length of the snake (P = 0.017), venom antigenaemia (P = 0.016), initial grading (P < 0.001), and with the need for surgical debridement (n = 10/70, P < 0.001). Outcome was correlated with initial severity of envenoming. However, the only factor with a positive prognostic significance for the individual envenomed patient was the early infusion of specific antivenom, which led to 100% recovery in our series. PMID:9861375

  18. A influência dos ritmos circadianos no metabolismo da serpente Bothrops jararaca (Serpentes, Viperidae - DOI: 10.4025/actascibiolsci.v30i3.5030 The influence of circadian rhythms on the metabolism of the snake Bothrops jararaca (Serpentes, Viperidae - DOI: 10.4025/actascibiolsci.v30i3.5030

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    José Geraldo Pereira da Cruz

    2008-10-01

    Full Text Available A atividade termorreguladora conduziu a uma busca extensiva para o entendimento das correlações entre as variáveis fisiológicas, incluindo as funções metabólicas e a temperatura corporal. Frequentes observações mostram que algumas serpentes podem se aquecer, sendo este aumento de temperatura independente da temperatura ambiente, indicando a termorregulação bem sucedida. Bothrops jararaca foram expostas a dois ambientes com diferentes temperaturas (20 e 30oC durante três semanas, sendo mensuradas a temperatura corporal e o consumo de oxigênio. O aumento da temperatura corporal e consumo de oxigênio de Bothrops jararaca ocorreram na fase de escuro do fotoperíodo, consistente para espécies noturnas. Entretanto, antecedendo a fase de escuro, as serpentes em 20oC apresentaram os níveis mais elevados durante o dia para temperatura corporal e consumo de oxigênio. Estes resultados indicam pela primeira vez que animais termodependentes podem controlar a temperatura corporal por meio de ritmos fisiológicos circadianos, semelhante aos observados em termoindependentes. Os ritmos circadianos permitem que os animais antecipem as mudanças no ambiente: parâmetros fisiológicos como a temperatura corporal e as reservas de energia ou sua mobilização podem ser ajustadas antes que as mudanças ambientais previstas ocorram realmente.The thermoregulatory activity has led to an extensive search for correlations between physiological variables, including metabolic functions, and the ideal level of body temperature. Snakes were also often seen basking, when their body temperatures were relatively independent of ambient temperature, indicating successful thermoregulation. Bothrops jararaca were exposed to two different ambient temperatures (20 and 30ºC over a time course of three weeks and oxygen consumption and body temperature were measured. The snakes exhibited a freerunning rhythm of body temperature. Metabolic rate was increased at the same

  19. Hemiparesia esquerda consecutiva a empeçonhamento: Por Bothrops Jararacussu

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    Napoleão L. Teixeira

    1944-09-01

    Full Text Available O A. apresenta uma observação de hemiparesia esquerda, consecutiva a envenenamento por Bothrops Jararacussu. É o segundo caso que publica, com idêntica etiologia; além das suas, há duas outras observações semelhantes, de Octávio de Magalhães. Depois de discorrer sobre as hemiplegias cerebrais infantis - mais comuns - lembra que estas nem sempre se acompanham de espasticidade, podendo apresentar-se, raramente, com hipotonia muscular, ou mesmo de coréia, atetose, ou movimentos córeo-atetóticos que falam em favor de comprometimento extrapiramidal. Assinala, também, que crises epileptiformes - generalizadas ou parciais - podem ocorrer, o mesmo se podendo dizer de distúrbios psíquicos. Traduzir-se-iam, estes, por déficit intelectual marcado, podendo ir desde a debilidade mental até a idiotia; ou então, mesmo quando não há aparente comprometimento da esfera intelectual, por perturbações da afetividade, da vontade e do pragmatismo. Passando em revista as causas mais frequentes das hemiplegias cerebrais infantis, abre lugar, dentre elas, para os empeçonhamentos ofídico e escorpiônico (Magalhães e Guimarães. Estuda, a seguir, a patogenia das hemiplegias no decurso dos referidos empeçonhamentos, alinhando as hipóteses correntes, da preferência de conceituadas autoridades no assunto. Transcreve, finalmente, sua observação clínica, da qual se pode concluir, sem dúvida possível, haver, no caso "que apresenta, estreita e imediata ligação entre o envenenamento ofídico e a hemiparesia em estudo. O observado seria portador, além do mais, de crises epileptiformes generalizadas, aparecidas logo a seguir ao empeçonhamento. Além da fotografia do caso que apresenta, mostra-nos o A. outra, de caso anteriormente publicado, este, porém, de hemiplegia cerebral infantil típica, instalada aos 11 anos, após picada de Bothrops jararaca.

  20. IgG antibodies against phospholipase A2 from Crotalus durissus terrificus: cross-reaction with venoms from Bothrops species from Argentina

    OpenAIRE

    JP Rodríguez; MC De Marzi; S Maruñak; P Teibler; O Acosta; EL Malchiodi; LC Leiva

    2009-01-01

    We examined the ability of IgG anti-crotalic PLA2 to cross-react with Bothrops spp. venoms, from snakes found in the northeast of Argentina. Immunoblotting and ELISA tests showed that IgG anti-crotalic PLA2 recognize antigens of bothropic venoms. Indirect hemolytic activity tests showed that the quantity of antibodies that neutralized 50% of Crotalus durissus terrificus venom (ED50: 2.1 mg IgG anti-crotalic PLA2/100 µg of venom) were also able to neutralize venom from other snakes in the foll...

  1. Envenenamento experimental por Bothropoides jararaca e Bothrops jararacussu em ovinos: aspectos clínico-patológicos e laboratoriais Experimental poisoning by Bothropoides jararaca and Bothrops jararacussu in sheep: clinic-pathological and laboratory aspects

    Directory of Open Access Journals (Sweden)

    Ana Paula Aragão

    2010-09-01

    Bothropoides jararaca and Bothrops jararacussu venom to provide subsidies for the differential diagnosis of snake bites. The liofilized venoms were diluted in 1 ml saline and administrated subcutaneously to four sheep. Three of the animals died, and the one that received 0.5mg/kg (B. jararaca venom recovered. First symptoms were observed from 7 minutes to 1 hour after inoculation, and the clinical course varied from 7 hours and 9 minutes to 21 hours and 59 minutes. The symptoms, independent of the dosage, were swelling of the inoculation site, increased bleeding time and capillary filling, tachycardia, dyspnea, pale mucous membranes and diminished reaction to external stimuli. Laboratory tests revealed pronounced normocytic and normochromic anemia, trombocytopenia, slight reduction of fibrogen and total plasmatic protein, in two animals diminished hematocrit, besides pronounced increase of creatinaquinase and lactic dehydrogenase. At necropsy, the main findings at the inoculation site and adjacent tissues were extensive hemorrhages in the sheep inoculated with jararaca venom, and predominantly edema in the two animals inoculated with jararacussu venom. In two sheep which received jararacussu venom, acute pulmonary edema was observed. Hemorrhage and edema as the main histopathological changes, besides necrosis of muscle fibers and vessels at the inoculation site and adjacent tissue was observed. The renal tubular necrosis was attributed to shock. The volume increase at the inoculation site and surroundings was mainly due to hemorrhage (B. jararaca or edema (B. jararacussu.

  2. Eficacia experimental de anticuerpos IgY producidos en huevos, contra el veneno de la serpiente peruana Bothrops atrox Experimental efficacy of IgY antibodies produced in eggs against the venom of the Peruvian snake Bothrops atrox

    OpenAIRE

    Julio C. Mendoza; Dan Vivas; Edith Rodríguez; Rosio Inga; Gustavo Sandoval; Fanny Lazo; Armando Yarlequé

    2012-01-01

    Objetivos. Desarrollar un protocolo de inmunización para producir inmunoglobulinas IgY de origen aviar contra el veneno de la serpiente peruana Bothrops atrox y evaluar la capacidad neutralizante. Materiales y métodos. Se inmunizaron seis gallinas de postura de la raza hy line brown con 500 μg/dosis de veneno de B. atrox en un periodo de dos meses. Cada semana, los huevos fueron colectados para el aislamiento de inmunoglobulinas IgY a partir de la yema, usando dos pasos consecutivos con ...

  3. The effects of low-level laser on muscle damage caused by Bothrops neuwiedi venom

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    DM Dourado

    2008-01-01

    Full Text Available The present study aimed to assess the effects of low-level laser (660 nm on myonecrosis caused by the insertion of Bothrops neuwiedi venom in the gastrocnemius muscle of rats. Male Wistar rats were divided into three groups (n = 24 each: Group S (0.9% saline solution; Group V (venom and Group VLLL (venom plus low-level laser. These categories were subdivided into four additional groups (n = 6 based on the euthanasia timing (3 hours, 24 hours, 3 days and 7 days. The groups V and VLLL were inoculated with 100 µL of concentrated venom (40 µg/mL in the gastrocnemius muscle. The muscle was irradiated using a gallium-aluminum-arsenide laser (GaAlAs at 35 mW power and 4 J/cm² energy density for 3 hours, 24 hours, 3 days or 7 days after venom inoculation. To evaluate the myotoxic activity of the venom, CK activity was measured and the muscle was histologically analyzed. The low-level laser reduced venom-induced CK activity in the groups euthanized at 3 hours, 24 hours and 3 days (p < 0.0001. Histological analysis revealed that low-level laser reduced neutrophilic inflammation as well as myofibrillar edema, hemorrhage and myonecrosis following B. neuwiedi envenomation. These results suggest that low-level laser can be useful as an adjunct therapy following B. neuwiedi envenomation.

  4. Envenomations by Bothrops and Crotalus snakes induce the release of mitochondrial alarmins.

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    Irene Zornetta

    Full Text Available Skeletal muscle necrosis is a common manifestation of viperid snakebite envenomations. Venoms from snakes of the genus Bothrops, such as that of B. asper, induce muscle tissue damage at the site of venom injection, provoking severe local pathology which often results in permanent sequelae. In contrast, the venom of the South American rattlesnake Crotalus durissus terrificus, induces a clinical picture of systemic myotoxicity, i.e., rhabdomyolysis, together with neurotoxicity. It is known that molecules released from damaged muscle might act as 'danger' signals. These are known as 'alarmins', and contribute to the inflammatory reaction by activating the innate immune system. Here we show that the venoms of B. asper and C. d. terrificus release the mitochondrial markers mtDNA (from the matrix and cytochrome c (Cyt c from the intermembrane space, from ex vivo mouse tibialis anterior muscles. Cyt c was released to a similar extent by the two venoms whereas B. asper venom induced the release of higher amounts of mtDNA, thus reflecting hitherto some differences in their pathological action on muscle mitochondria. At variance, injection of these venoms in mice resulted in a different time-course of mtDNA release, with B. asper venom inducing an early onset increment in plasma levels and C. d. terrificus venom provoking a delayed release. We suggest that the release of mitochondrial 'alarmins' might contribute to the local and systemic inflammatory events characteristic of snakebite envenomations.

  5. The effects of low-level laser on muscle damage caused by Bothrops neuwiedi venom

    International Nuclear Information System (INIS)

    The present study aimed to assess the effects of low-level laser (660 nm) on myonecrosis caused by the insertion of Bothrops neuwiedi venom in the gastrocnemius muscle of rats. Male Wistar rats were divided into three groups (n = 24 each): Group S (0.9% saline solution); Group V (venom) and Group VLLL (venom plus low-level laser). These categories were subdivided into four additional groups (n = 6) based on the euthanasia timing (3 hours, 24 hours, 3 days and 7 days). The groups V and VLLL were inoculated with 100 μL of concentrated venom (40 μg/mL) in the gastrocnemius muscle. The muscle was irradiated using a gallium-aluminum-arsenide laser (GaAlAs) at 35 mW power and 4 J/cm2 energy density for 3 hours, 24 hours, 3 days or 7 days after venom inoculation. To evaluate the myotoxic activity of the venom, CK activity was measured and the muscle was histologically analyzed. The low-level laser reduced venom-induced CK activity in the groups euthanized at 3 hours, 24 hours and 3 days (p < 0.0001). Histological analysis revealed that low-level laser reduced neutrophilic inflammation as well as myofibrillar edema, hemorrhage and myonecrosis following B. neuwiedi envenomation. These results suggest that low-level laser can be useful as an adjunct therapy following B. neuwiedi envenomation. (author)

  6. Bothrops jararacussu venom-induced neuromuscular blockade inhibited by Casearia gossypiosperma Briquet hydroalcoholic extract

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    TM Camargo

    2010-01-01

    Full Text Available The hydroalcoholic extract of Casearia gossypiosperma Briquet (Flacourtiaceae was standardized for the first time through quality control procedures including pharmacognostic methods, fingerprint chromatograms, defined amounts of marker substances and physicochemical characteristics. The pharmacological activity of C. gossypiosperma (Cg hydroalcoholic extract was assayed by a traditional in vitro test, which involved irreversible neuromuscular blockade induced by Bothrops jararacussu (Bjssu venom (60 µg/mL in mouse phrenic nerve-diaphragm preparations. Bjssu venom blocked muscle activity for 26 (± 2.0 minutes (n = 6. Cg extract (0.1 mg/mL induced changes on the baseline muscle activity without impairing the muscle function and inhibited 87.6% (± 1.8 (n = 6 of the Bjssu venom-induced blockade. Both flavonoids (0.624 g% and polyphenols (4.63 g% from the extract were spectrophotometrically quantified. Therefore, the present study confirms the antibothropic activity of Cg extract, supporting the ethnomedical use of Casearia sp. in the treatment of snakebite victims.

  7. Purification and functional characterization of a new metalloproteinase (BleucMP) from Bothrops leucurus snake venom.

    Science.gov (United States)

    Gomes, Mário Sérgio R; de Queiroz, Mayara R; Mamede, Carla C N; Mendes, Mirian M; Hamaguchi, Amélia; Homsi-Brandeburgo, Maria I; Sousa, Marcelo V; Aquino, Elaine Nascimento; Castro, Mariana S; de Oliveira, Fábio; Rodrigues, Veridiana M

    2011-04-01

    A fibrino(geno)lytic nonhemorrhagic metalloproteinase (BleucMP) was purified from Bothrops leucurus snake venom by two chromatographic steps procedure on DEAE-Sephadex A-25 followed by CM-Sepharose Fast Flow column. BleucMP represented 1.75% (w/w) of the crude venom and was homogeneous on SDS-PAGE. BleucMP analyzed by MALDI TOF/TOF, showed a molecular mass of 23,057.54Da and when alkylated and reduced, the mass is 23,830.40Da. Their peptides analyzed in MS (MALDI TOF\\TOF) showed significant score when compared with those of other proteins by NCBI-BLAST2 alignment display. As regards their proteolytic activities, BleucMP efficiently acted on fibrinogen, fibrin, and was inhibited by EDTA and 1.10-phenanthroline. This enzyme was also able to decrease significantly the plasma fibrinogen level provoking blood incoagulability, however was devoid of hemorrhagic activity when tested in the mice skin and did not induce relevant biochemical, hematological and histopathological alterations in mice. The aspects addressed in this paper provide data on the effect of BleucMP in envenomation from B. leucurus snakes in order to better understand the effects caused by snake venom metalloproteinase. PMID:21130897

  8. The effects of low-level laser on muscle damage caused by Bothrops neuwiedi venom

    Energy Technology Data Exchange (ETDEWEB)

    Dourado, D.M.; Matias, R.; Almeida, M.F.; Paula, K.R. de; Carvalho, P.T.C. [University for the Development of the State and of the Region of Pantanal (UNIDERP), Campo Grande, MS (Brazil). Lab. of Experimental Histopathology]. E-mail: ccfi@uniderp.br; Vieira, R.P. [University of Sao Paulo (USP), SP (Brazil). School of Medicine. Dept. of Pathology and Physical Therapy; Oliveira, L.V.F. [Nove de Julho University (UNINOVE), Sao Paulo, SP (Brazil). Masters Program in Rehabilitation Sciences

    2008-07-01

    The present study aimed to assess the effects of low-level laser (660 nm) on myonecrosis caused by the insertion of Bothrops neuwiedi venom in the gastrocnemius muscle of rats. Male Wistar rats were divided into three groups (n = 24 each): Group S (0.9% saline solution); Group V (venom) and Group VLLL (venom plus low-level laser). These categories were subdivided into four additional groups (n = 6) based on the euthanasia timing (3 hours, 24 hours, 3 days and 7 days). The groups V and VLLL were inoculated with 100 {mu}L of concentrated venom (40 {mu}g/mL) in the gastrocnemius muscle. The muscle was irradiated using a gallium-aluminum-arsenide laser (GaAlAs) at 35 mW power and 4 J/cm{sup 2} energy density for 3 hours, 24 hours, 3 days or 7 days after venom inoculation. To evaluate the myotoxic activity of the venom, CK activity was measured and the muscle was histologically analyzed. The low-level laser reduced venom-induced CK activity in the groups euthanized at 3 hours, 24 hours and 3 days (p < 0.0001). Histological analysis revealed that low-level laser reduced neutrophilic inflammation as well as myofibrillar edema, hemorrhage and myonecrosis following B. neuwiedi envenomation. These results suggest that low-level laser can be useful as an adjunct therapy following B. neuwiedi envenomation. (author)

  9. Effects of photobiostimulation on edema and hemorrhage induced by Bothrops moojeni venom.

    Science.gov (United States)

    Nadur-Andrade, Nikele; Barbosa, Ana Maria; Carlos, Fernando Pereira; Lima, Carlos José; Cogo, José Carlos; Zamuner, Stella Regina

    2012-01-01

    Antivenom (AV) treatment has been ineffective in neutralizing the severe local fast-developing tissue damage following snake-bite envenoming. We studied the effectiveness of low-level laser (LLL) and light-emitting diode (LED) irradiation alone or in combination with AV in reducing local edema formation and hemorrhage induced by Bothrops moojeni venom (BmV) in mice. Edema formation was induced by injection of 1 μg per paw of BmV into the right paw and was evaluated before and at several intervals after BmV intraplantar injection. Hemorrhagic activity was evaluated after intradermal injection of 20 μg of BmV by measuring the diameter of the hemorrhagic area on the inner side of the skin. The site of BmV injection was irradiated by LLL or LED 30 min after BmV inoculation. AV was also administered intravenously 30 min after BmV injection. Irradiation with LLL at a wavelength of 685 nm and a dose of 2.2 J/cm(2) and with a red LED and an infrared LED at wavelengths of 635 nm and 945 nm, respectively, and a dose of 4 J/cm(2) reduced edema formation and hemorrhage induced by BmV (p BmV may be clinically relevant.

  10. Evaluation of anti-Bothrops asper venom activity of ethanolic extract of Brownea rosademonte leaves

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    Salazar Marcos

    2014-12-01

    Full Text Available Significant inhibition of the coagulant and hemorrhagic effects of Bothrops asper venom was demonstrated by ethanolic extract prepared from the leaves of Brownea rosademonte. In vitro experiments preincubating 5.5 mg of extract kg-1 b.m. for 30 min with a minimum hemorrhagic dose of venom (273.8 ± 16.1 μg of venom kg-1 b.m. lowered the hemorrhagic activity of the venom alone in CD-1 mice by 51.5 ± 2.6 %. Additionally, 1.7 mg extract L-1 plasma prolonged 5.1 times the plasma coagulation time. Fractionation of the extract led to the isolation of two compounds: ononitol (1 and quercetrin (2. The structure of compounds 1 and 2 was established by spectroscopic analyses, including APCI-HRMS and NMR (1H, 13C, HSQC, HMBC and COSY. A quercetrin concentration of 0.11 μmol L-1 prolonged the plasma coagulation time 2.6 times demonstrating that this compound was one of the active constituents of the Brownea rosademonte extract.

  11. Pulsed ultrasound therapy accelerates the recovery of skeletal muscle damage induced by Bothrops jararacussu venom

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    J. Saturnino-Oliveira

    2012-06-01

    Full Text Available We studied the effect of pulsed ultrasound therapy (UST and antibothropic polyvalent antivenom (PAV on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group received a perimuscular injection of venom (1 mg/kg and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm², pulsed mode. Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively. Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively. The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.

  12. Acción de la miotoxina del veneno de Bothrops brazili Hoge, 1953 (Ophidia: Viperidae

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    Carmen Pantigoso

    2013-06-01

    Full Text Available Se ha estudiado el modo de acción de la miotoxina aislada del veneno de la serpiente Bothrops brazili. La inoculación de la miotoxina en el músculo gastrocnemius de ratones albinos produce durante la primera hora de acción la liberación de creatina kinasa y lactato deshidrogenasa, mientras que por PAGE-SDS, se revela que la incubación de la miotoxina con músculo gastrocnemius aislado, produce además la liberación de otras proteínas musculares. Asimismo, la miotoxina produce hipercontracción, lesiones delta e incrementa los niveles de calcio intramuscular, tanto in vivo como in vitro, lo cual no depende del ingreso de calcio extracelular vía receptores de dihidropiridina. Este incremento de calcio explicaría la hipercontracción observada y podría generar la activación de proteasas y lipasas endógenas dependientes de calcio, que conducirían a la necrosis muscular.

  13. Coagulopathy after snake bite by Bothrops neuwiedi: case report and results of in vitro experiments.

    Science.gov (United States)

    Dempfle, C E; Kohl, R; Harenberg, J; Kirschstein, W; Schlauch, D; Heene, D L

    1990-12-01

    Coagulation studies were performed in a patient who had been bitten by a snake of the species Bothrops neuwiedi. The patient presented with hemorrhagic necrosis at the envenomization site and considerable bleeding from venous puncture sites. He developed a severe defibrination syndrome with a clottable fibrinogen level of approximately 0.1 g/l. Fibrinogen was not measurable by clotting time assay. Fibrin degradation products were greatly elevated. Treatment with antivenom caused an anaphylactic reaction within ten minutes and serum sickness after three days. In vitro experiments revealed that B. neuwiedi venom directly activates Factors II and X, but does not activate Factor XIII. In vivo consumption of Factor XIII after B. neuwiedi envenomization is ascribed to the action of Factor IIa. At low venom concentrations clotting is initiated by activation of prothrombin by the venom either directly or via Factor X activation. Treatment with heparin might be beneficial in coagulopathy secondary to snake bite by reducing circulating active thrombin. The venom contains thrombin-like proteases which cause slow clotting of fibrinogen, and plasmin-like components causing further proteolysis of fibrinogen and fibrin. Antivenom has no effect on the proteolytic action of the snake venom. The in vivo effects of antivenom are presumably caused by acceleration of the elimination of venom components from the circulation. Intravenous administration of antivenom caused normalization of blood coagulation parameters within 48 h. PMID:2291986

  14. Two related thrombin-like enzymes present in Bothrops atrox venom

    Directory of Open Access Journals (Sweden)

    J.H. Petretski

    2000-11-01

    Full Text Available This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical Km on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs raised against the purified enzyme forms recognized different epitopes of the putative corresponding enzymes present in B. atrox crude venom. On Western blotting analysis of B. atrox crude venom, mAbs 5DB2C8, 5AA10 and 5CF11, but not mAbs 6CC5 and 6AD2-G5, revealed two or more protein bands ranging from 25 to 38 kDa. By immunoprecipitation assays, the 6AD2-G5 mAb was able to precipitate protein bands of 36-38 kDa from B. atrox, B. leucurus, B. pradoi, B. moojeni, B. jararaca and B. neuwiedii crude venoms. Fibrinogen-clotting activity was inhibited when the same venom specimens were pre-incubated with mAb 6AD2-G5, except for B. jararaca and B. neuwiedii.

  15. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom.

    Science.gov (United States)

    Mendonça-Franqueiro, Elaine de Paula; Alves-Paiva, Raquel de Melo; Sartim, Marco Aurélio; Callejon, Daniel Roberto; Paiva, Helder Henrique; Antonucci, Gilmara Ausech; Rosa, José César; Cintra, Adélia Cristina Oliveira; Franco, João José; Arantes, Eliane Candiani; Dias-Baruffi, Marcelo; Sampaio, Suely Vilela

    2011-03-01

    Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycan-binding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability, which suggested that galatrox might contribute to venom toxicity by directly inducing cell death. PMID:21297119

  16. A rational protocol for the successful crystallization of L-amino-acid oxidase from Bothrops atrox.

    Science.gov (United States)

    Alves, Raquel Melo; Feliciano, Patricia Rosa; Sampaio, Suely Vilela; Nonato, Maria Cristina

    2011-04-01

    Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme L-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 Å resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 Å, β = 96.03°. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V(M) of 2.12 Å(3) Da(-1), corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques. PMID:21505245

  17. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom

    Institute of Scientific and Technical Information of China (English)

    Elaine de Paula Mendonca-Franqueiro; Eliane Candiani Arantes; Marcelo Dias-Baruffi; Suely Vilela Sampaio; Raquel de Melo Alves-Paiva; Marco Aurélio Sartim; Daniel Roberto Callejon; Helder Henrique Paiva; Gilmara Ausech Antonucci; José César Rosa; Adélia Cristina Oliveira Cintra; Jo(a)o José Franco

    2011-01-01

    Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycanbinding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability,which suggested that galatrox might contribute to venom toxicity by directly inducing cell death.

  18. Hyperbaric oxygen therapy after Bothrops lanceolatus snake bites in Martinique: a brief report.

    Science.gov (United States)

    Hochedez, P; Thomas, L; Mehdaoui, H

    2010-01-01

    Every year 10 to 20 cases of snake bites are reported on the Caribbean island of Martinique. The only snake involved, Bothrops lanceolatus, is endemic on the island, and its bite may lead to systemic multifocal thrombotic complications in the'absence of the monospecific antivenom. Between January 1988 and January 2009, more than 250 snake bites have been reported, and five patients were treated with hyperbaric oxygen (HBO2) therapy for local complications. The patients were male, bitten on the leg or the hand, and presented with severe complications such as necrotizing soft tissue infections, compartment syndrome or abscesses despite prompt wound care and administration of antivenomous serum. Outcomes were favorable for these five patients, except for one who was left with a functional defect of the hand. Although snake bites are not part of the currently recommended indications for HBO2 therapy, local complications, namely compartment syndrome, necrotizing soft tissue infections and enhancement of healing in selected problem wounds, are approved uses of HBO2 therapy as defined by the Hyperbaric Oxygen Therapy Committee and would benefit from prospective studies. PMID:21226390

  19. Coagulant and anticoagulant activities of Bothrops lanceolatus (Fer de lance) venom.

    Science.gov (United States)

    Lôbo de Araújo, A; Kamiguti, A; Bon, C

    2001-01-01

    Bothrops lanceolatus venom contains caseinolytic, phospholipase, esterase and haemorrhagic activities. We have investigated the coagulant and anticoagulant actions of B. lanceolatus venom on human citrated plasma and on purified plasma components. Although B. lanceolatus venom up to 50 microg/ml was unable to clot citrated plasma, at concentrations > or = 5 microg/ml the venom dose-dependently clotted purified human fibrinogen, indicating the presence of a thrombin-like enzyme. Human plasma (final concentration > or = 12.5%) dose-dependently inhibited the venom-induced fibrinogen clotting. This finding suggested that endogenous plasma protease inhibitors can affect the venom's action on fibrinogen. To investigate this possibility, B. lanceolatus venom was incubated with different plasma protease inhibitors and the activity on fibrinogen tested. alpha(2)-Macroglobulin and alpha(1)-antitrypsin did not interfere with the coagulant activity of the venom whereas the antithrombin-III/heparin complex partially inhibited this activity. A non-toxic, acidic phospholipase A(2) purified from B. lanceolatus venom prolonged the activated partial thromboplastin time in human plasma from 39.7+/-0.5 s (control with saline) to 60.2+/-0.9 s with 50 microg of PLA(2) (p<0.001), suggesting an anticoagulant activity associated with this enzyme. This anticoagulant activity may account for some of the effects of the venom on blood coagulation. PMID:10978756

  20. Expression of Muscle LIM Protein Gene from Monochamus alternatus in Response to 11 Kinds of Pesticides%11种杀虫剂对松墨天牛肌肉LIM蛋白基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    罗淋淋; 林同

    2014-01-01

    松墨天牛(Monochamusalternatus Hope)是松树的重要蛀干害虫,也是松材线虫(Bursaphelenchusxylophilus)的主要传播媒介。试验从松墨天牛文库中分离得到肌肉LIM蛋白基因的部分cDNA序列,长度为871 bp,命名为MaLIM(GenBank:KJ872589);与赤拟谷盗(Tribolium castaneum)和家蚕(Bombyxmori)氨基酸相似性分别为77%和75%。用RT-qPCR测定了11种杀虫剂对MaLIM相对表达量的影响,结果表明,杀虫剂处理后MaLIM基因的表达量均有所下降,仅为对照的0.03~0.53。%Monochamus alternatus H ope is one of the important pests of pine stemborers,and also is vector of pine wood nematode (Bursaphelenchusxylophilus).Inthisstudy,apartialcDNA sequenceofmuscleLIMproteingene with870bpinlengthwasisolatedfromM.alternatuslibrary and named MaLIM(G enB ank:K J872589) thathas 77% and 75% amino acid similarity with the muscle L IMprotein of Triboliumcastaneumand Bombyx mori respectively. T he relative expression of MaLIMin response to 11 kinds of insecticides was determined by R T-qPC R ,the results showed thatthe MaLIMwas down regulated and the expression levelwas changed from0.03 times to 0.53 times of the control.

  1. Low-level laser therapy decreases local effects induced by myotoxins isolated from Bothrops jararacussu snake venom

    OpenAIRE

    AM Barbosa; AB Villaverde; L Guimarães-Sousa; AM Soares; SF Zamuner; JC Cogo; SR Zamuner

    2010-01-01

    The prominent myotoxic effects induced by Bothrops jararacussu crude venom are due, in part, to its polycationic myotoxins, BthTX-I and BthTX-II. Both myotoxins have a phospholipase A2 structure: BthTX-II is an active enzyme Asp-49 PLA2, while BthTX-I is a Lys-49 PLA2 devoid of enzymatic activity. In this study, the effect of low-level laser therapy (LLLT), 685 nm laser at a dose of 4.2 J/cm2 on edema formation, leukocyte influx and myonecrosis caused by BthTX-I and BthTX-II, isolated from Bo...

  2. Inhibition of the hyperalgesic activity of Bothrops jararaca venom by an antibothropic fraction isolated from opossum (Didelphis marsupialis) serum.

    Science.gov (United States)

    Rocha, S L; Frutuoso, V S; Domont, G B; Martins, M A; Moussatché, H; Perales, J

    2000-06-01

    The antibothropic fraction (ABF) already isolated from Didelphis marsupialis serum, inhibits the haemorrhagic, oedematogenic, myonecrotic and lethal activities of Bothrops jararaca venom (Bjv). The aim of this work was to verify the capability of ABF to inhibit the hyperalgesic activity of Bjv. Intraplantar injection of Bjv induced hyperalgesia in a time- and dose-dependent manner and ABF administered in situ concomitantly with Bjv or i.v. 30 min before venom injection reduced the induced hyperalgesia. This same effect was observed when ABF was intravenously injected at 5 and 15 min after Bjv. Our results show that ABF inhibits also the hyperalgesia induced by Bjv. PMID:10695972

  3. Comparative evaluation of adverse effects in the use of powder trivalent antivenom and liquid antivenoms in Bothrops snake bites

    Directory of Open Access Journals (Sweden)

    Iran Mendonça da Silva

    2012-08-01

    Full Text Available INTRODUCTION: Snake bite, a problem in public health, generally occurs where there is no electric power. METHODS: A comparative clinical study was conducted with 102 victims of Bothrops snake bite, from the state of Amazonas, Brazil; 58 victims were treated with liofilizated trivalent antivenom serum (SATL and 44 victims treated with liquid bivalent and monovalent antivenom serum (SAMBL. RESULTS: 17% (10/58 of patients presented adverse effects with the SATL and 25% (11/44 with the SAMBL. CONCLUSIONS: There was no statistic difference in number of adverse effects between the two types of snake bite antivenom.

  4. N-terminal amino acid sequences and some characteristics of fibrinolytic/hemorrhagic metalloproteinases purified from Bothrops jararaca venom.

    Science.gov (United States)

    Maruyama, Masugi; Sugiki, Masahiko; Anai, Keita; Yoshida, Etsuo

    2002-08-01

    We determined the N-terminal amino acid sequences of the fibrinolytic/hemorrhagic metalloproteinases (jararafibrases I, III and IV) purified from Bothrops jararaca venom. The N-terminal amino acid sequences of jararafibrase I and its degradation products were identical to those of jararhagin, another hemorrhagic metalloproteinase purified from the same snake venom. Together with enzymatic and immunological properties, we concluded that those two enzymes are identical. The N-terminal amino acid sequence of jararafibrase III was quite similar to C-type lectin isolated from Crotalus atrox, and the protein had a hemagglutinating activity on intact rat red blood cells. PMID:12165326

  5. Neutralization of four Peruvian Bothrops sp. snake venoms by polyvalent antivenoms produced in Perú and Costa Rica: preclinical assessment.

    Science.gov (United States)

    Rojas, Ermila; Quesada, Lil; Arce, Viviana; Lomonte, Bruno; Rojas, Gustavo; Gutiérrez, José María

    2005-01-01

    Envenomations after bites inflicted by snakes of the genus Bothrops constitute a public health hazard in Perú, and the intravenous administration of equine-derived antivenoms represents the only scientifically validated treatment. This study presents a preclinical assessment of the efficacy of two whole IgG antivenoms, prepared in Perú and Costa Rica, to neutralize the most relevant toxic effects induced by the venoms of Bothrops atrox, B. brazili, B. barnetti and B. pictus from Perú. Peruvian antivenom is produced by immunizing horses with Bothrops sp. venoms from this country, whereas the production of Costa Rican antivenom involves immunization with venoms from Central American snakes. The neutralization of lethal, hemorrhagic, edema-forming, myotoxic, coagulant and defibrinating activities was evaluated in assays involving incubation of venom and antivenom prior to testing. Both antivenoms were effective in the neutralization of these effects, with quantitative variations in the values of effective dose 50% depending on the effects being studied. Peruvian antivenom was more effective in the neutralization of lethality induced by B. atrox and B. barnetti venoms. However, Peruvian antivenom failed to neutralize coagulant activity of B. barnetti venom and edema-forming activity of B. brazili venom, whereas neutralization was achieved by Costa Rican antivenom. It is concluded that an extensive immunological cross-reactivity exists between Bothrops sp. venoms from Perú and Costa Rica, and that both antivenoms are effective in the neutralization of these four venoms in a rodent model of envenoming. PMID:15589801

  6. Eficacia experimental de anticuerpos IgY producidos en huevos, contra el veneno de la serpiente peruana Bothrops atrox Experimental efficacy of IgY antibodies produced in eggs against the venom of the Peruvian snake Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Julio C. Mendoza

    2012-03-01

    Full Text Available Objetivos. Desarrollar un protocolo de inmunización para producir inmunoglobulinas IgY de origen aviar contra el veneno de la serpiente peruana Bothrops atrox y evaluar la capacidad neutralizante. Materiales y métodos. Se inmunizaron seis gallinas de postura de la raza hy line brown con 500 μg/dosis de veneno de B. atrox en un periodo de dos meses. Cada semana, los huevos fueron colectados para el aislamiento de inmunoglobulinas IgY a partir de la yema, usando dos pasos consecutivos con αcido caprνlico y sulfato de amonio. La detecciσn de anticuerpos se realizσ por inmunodifusiσn doble mientras que el tνtulo y reactividad cruzada se determinaron por las técnicas de ELISA y Western blot. El cálculo de DL50 y de la DE50 del antiveneno IgY producido se realizó utilizando el método de Probits. Resultados. La masa de anticuerpos aislados fue de 8,5 ± 1,35 mg de IgY/mL de yema. Asimismo, la DE50 del antiveneno aviar fue calculada en 575 μL de antiveneno/mg de veneno. Adicionalmente, los ensayos de reactividad cruzada mostraron que el veneno de B. atrox comparte mas epνtopes comunes con el veneno de B. brazili (47% que con otros veneno del mismo género, en tanto que los venenos de Lachesis muta (19% y Crotalus durissus (12% mostraron una baja reactividad cruzada. Conclusiones. Se ha obtenido IgY purificada contra el veneno de B. atrox con capacidad neutralizante y se ha demostrado su utilidad como herramienta inmunoanalítica para evaluar la reactividad cruzada con venenos de otras especies.Objectives. To develop an immunization protocol in order to produce avian IgY immunoglobulins against Bothrops atrox Peruvian snake venom and to evaluate its neutralizing capacity. Materials and methods. Six Hy Line Brown hens were immunized each two weeks using 500μg/doses of B. atrox venom in a period of two months. Each week, eggs were collected for IgY isolation from yolk using two consecutive steps with caprilic acid and ammonium sulfate

  7. Evaluation of platelet number and function and fibrinogen level in patients bitten by snakes of the Bothrops genus

    Directory of Open Access Journals (Sweden)

    Fábio Cardoso Luan

    1995-03-01

    Full Text Available Platelet function and plasma fibrinogen levels were evaluated in 14 patients, 10 males and 4females, aged 13-59years bitten by Bothrops genus snakes. There was a statistical difference (p Foram avaliadas a função plaquetária e os níveis séricos de fibrinogênio em 14 doentes picados por serpentes do gênero Bothrops, sendo 10 do sexo masculino e 4 do sexo feminino, com idades compreendidas entre 13 e 59 anos. Houve diferença estatística (p < 0,05 entre os níveis séricos defibrinogênio avaliados 24 e 48 horas após o acidente. Houve tendência à normalização após 48 horas do tratamento. A plaquetopenia foi evidente nas avaliações de 24 e 48 horas. Houve tendência à nomalização no 8o dia após o tratamento (p <0,05. Os níveis de produtos de degradação defibrina (PDF mostraram-se alterados em 83,33 % dos pacientes avaliados. Os autores sugerem que a hipoagregação esteja relacionada com níveis baixos de fibrinogênio e elevados de PDF.

  8. Purification and Biochemical Characterization of Three Myotoxins from Bothrops mattogrossensis Snake Venom with Toxicity against Leishmania and Tumor Cells

    Directory of Open Access Journals (Sweden)

    Andréa A. de Moura

    2014-01-01

    Full Text Available Bothrops mattogrossensis snake is widely distributed throughout eastern South America and is responsible for snakebites in this region. This paper reports the purification and biochemical characterization of three new phospholipases A2 (PLA2s, one of which is presumably an enzymatically active Asp49 and two are very likely enzymatically inactive Lys49 PLA2 homologues. The purification was obtained after two chromatographic steps on ion exchange and reverse phase column. The 2D SDS-PAGE analysis revealed that the proteins have pI values around 10, are each made of a single chain, and have molecular masses near 13 kDa, which was confirmed by MALDI-TOF mass spectrometry. The N-terminal similarity analysis of the sequences showed that the proteins are highly homologous with other Lys49 and Asp49 PLA2s from Bothrops species. The PLA2s isolated were named BmatTX-I (Lys49 PLA2-like, BmatTX-II (Lys49 PLA2-like, and BmatTX-III (Asp49 PLA2. The PLA2s induced cytokine release from mouse neutrophils and showed cytotoxicity towards JURKAT (leukemia T and SK-BR-3 (breast adenocarcinoma cell lines and promastigote forms of Leishmania amazonensis. The structural and functional elucidation of snake venoms components may contribute to a better understanding of the mechanism of action of these proteins during envenomation and their potential pharmacological and therapeutic applications.

  9. Isolation and Biochemical Characterization of a New Thrombin-Like Serine Protease from Bothrops pirajai Snake Venom

    Directory of Open Access Journals (Sweden)

    Kayena D. Zaqueo

    2014-01-01

    Full Text Available This paper presents a novel serine protease (SP isolated from Bothrops pirajai, a venomous snake found solely in Brazil that belongs to the Viperidae family. The identified SP, named BpirSP-39, was isolated by three chromatographic steps (size exclusion, bioaffinity, and reverse phase chromatographies. The molecular mass of BpirSP-39 was estimated by SDS-PAGE and confirmed by mass spectrometry (39,408.32 Da. The protein was able to form fibrin networks, which was not observed in the presence of serine protease inhibitors, such as phenylmethylsulfonyl fluoride (PMSF. Furthermore, BpirSP-39 presented considerable thermal stability and was apparently able to activate factor XIII of the blood coagulation cascade, unlike most serine proteases. BpirSP-39 was capable of hydrolyzing different chromogenic substrates tested (S-2222, S-2302, and S-2238 while Cu2+ significantly diminished BspirSP-39 activity on the three tested substrates. The enzyme promoted platelet aggregation and also exhibited fibrinogenolytic, fibrinolytic, gelatinolytic, and amidolytic activities. The multiple alignment showed high sequence similarity to other thrombin-like enzymes from snake venoms. These results allow us to conclude that a new SP was isolated from Bothrops pirajai snake venom.

  10. Exploring the proteomes of the venoms of the Peruvian pit vipers Bothrops atrox, B. barnetti and B. pictus.

    Science.gov (United States)

    Kohlhoff, Markus; Borges, Marcia H; Yarleque, Armando; Cabezas, Cesar; Richardson, Michael; Sanchez, Eladio F

    2012-04-01

    We report the comparative proteomic characterization of the venoms of Bothrops atrox, B. barnetti and B. pictus. The venoms were subjected to RP-HPLC and the resulting fractions analyzed by SDS-PAGE. The proteins were cut from the gels, digested with trypsin and identified via peptide mass fingerprint and manual sequencing of selected peptides by MALDI-TOF/TOF mass spectrometry. Around 20-25 proteins were identified belonging to only 6-7 protein families. Metalloproteinases of the classes P-I and P-III were the most abundant proteins in all venoms (58-74% based on peak area A214 nm), followed by phospholipases-A(2) (6.4-14%), disintegrins (3.2-9%) and serine proteinases (7-11%), and some of these proteins occurred in several isoforms. In contrast cysteine-rich secretory proteins and L-amino acid oxidases appeared only as single isoforms and were found only in B. atrox and B. barnetti. C-type lectins were also detected in all venoms but at low levels (~ 5%). Furthermore, the venoms contain variable numbers of peptides (Bothrops species is in agreement with their pharmacological and pathological effects. PMID:22300577

  11. Isolation and biological characterization of Batx-I, a weak hemorrhagic and fibrinogenolytic PI metalloproteinase from Colombian Bothrops atrox venom.

    Science.gov (United States)

    Patiño, Arley C; Pereañez, Jaime A; Núñez, Vitelbina; Benjumea, Dora M; Fernandez, Maritza; Rucavado, Alexandra; Sanz, Libia; Calvete, Juan J

    2010-11-01

    A hemorrhagic metalloproteinase, named Batx-I, was isolated from the venom of Bothrops atrox specimens (from Southeastern Colombian region) by a combination of CM-Sephadex C25 ion-exchange and Affi-gel Blue affinity chromatographies. This enzyme accounts for about 45% of venom proteins, and it has an ESI-MS isotope-averaged molecular mass of 23296.2 Da and a blocked N-terminus. Two internal fragments sequenced by mass spectrometric analysis showed similarity to other SVMPs from Bothrops venoms. To investigate the possible participation of Batx-I in the envenomation pathophysiology, proteolytic, fibrinogenolytic, hemorrhagic, and other biological activities were evaluated. The minimal hemorrhagic dose obtained was 17 microg/20 g body weight. The enzyme showed proteolytic activity on azocasein, comparable with activity of BaP1. This activity was inhibited by EDTA and 1, 10 o-phenanthroline but not by aprotinin, pepstatin A or PMSF. Fibrinogenolytic activity was analyzed by SDS-PAGE, revealing a preference for degrading the A alpha- and B beta-chains, although partial degradation of the gamma-chain was also detected. The protein lacks coagulant and defibrinating activity. The CK levels obtained, clearly reflects a myotoxic activity induced by Batx-I. The hemorrhagic and fibrinogenolytic activities exhibited by the isolated PI-SVMP may play a role in the hemorrhagic and blood-clotting disorders observed in patients bitten by B. atrox in Colombia. PMID:20600221

  12. Spectroscopic and thermal characterization of alternative model biomembranes from shed skins of Bothrops jararaca and Spilotis pullatus

    Directory of Open Access Journals (Sweden)

    André Rolim Baby

    2009-09-01

    Full Text Available Recently, there has been an interest in the use of shed snake skin as alternative model biomembrane for human stratum corneum. This research work presented as objective the qualitative characterization of alternative model biomembranes from Bothrops jararaca and Spilotis pullatus by FT-Raman, PAS-FTIR and DSC. The employed biophysical techniques permitted the characterization of the biomembranes from shed snake skin of B. jararaca and S. pullatus by the identification of vibrational frequencies and endothermic transitions that are similar to those of the human stratum corneum.Existe atualmente interesse no uso da muda de pele de cobra como modelos alternativos de biomembranas da pele humana. O presente trabalho apresentou como objetivo a caracterização qualitativa de modelos alternativos de biomembranas provenientes de mudas de pele de cobra da Bothrops jararaca e Spilotis pullatus por espectroscopia Raman (FT-Raman, espectroscopia fotoacústica no infravermelho (PAS-FTIR e calorimetria exploratória diferencial (DSC. As técnicas biofísicas FT-Raman, PAS-FTIR e DSC permitiram caracterizar qualitativamente os modelos alternativos de biomembranas provenientes das mudas de pele de cobra da B. jararaca e S. pullatus e identificar freqüências vibracionais e transições endotérmicas similares ao estrato córneo humano.

  13. Effects of Bothrops asper snake venom on lymphatic vessels: insights into a hidden aspect of envenomation.

    Directory of Open Access Journals (Sweden)

    Javier Mora

    Full Text Available BACKGROUND: Envenomations by the snake Bothrops asper represent a serious medical problem in Central America and parts of South America. These envenomations concur with drastic local tissue pathology, including a prominent edema. Since lymph flow plays a role in the maintenance of tissue fluid balance, the effect of B. asper venom on collecting lymphatic vessels was studied. METHODOLOGY/PRINCIPAL FINDINGS: B. asper venom was applied to mouse mesentery, and the effects were studied using an intravital microscopy methodology coupled with an image analysis program. B. asper venom induced a dose-dependent contraction of collecting lymphatic vessels, resulting in a reduction of their lumen and in a halting of lymph flow. The effect was reproduced by a myotoxic phospholipase A(2 (PLA(2 homologue isolated from this venom, but not by a hemorrhagic metalloproteinase or a coagulant thrombin-like serine proteinase. In agreement with this, treatment of the venom with fucoidan, a myotoxin inhibitor, abrogated the effect, whereas no inhibition was observed after incubation with the peptidomimetic metalloproteinase inhibitor Batimastat. Moreover, fucoidan significantly reduced venom-induced footpad edema. The myotoxic PLA(2 homologue, known to induce skeletal muscle necrosis, was able to induce cytotoxicity in smooth muscle cells in culture and to promote an increment in the permeability to propidium iodide in these cells. CONCLUSIONS/SIGNIFICANCE: Our observations indicate that B. asper venom affects collecting lymphatic vessels through the action of myotoxic PLA(2s on the smooth muscle of these vessels, inducing cell contraction and irreversible cell damage. This activity may play an important role in the pathogenesis of the pronounced local edema characteristic of viperid snakebite envenomation, as well as in the systemic biodistribution of the venom, thus representing a potential therapeutical target in these envenomations.

  14. A C-type lectin from Bothrops jararacussu venom disrupts Staphylococcal biofilms.

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    Raphael Contelli Klein

    Full Text Available Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16 showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model.

  15. Biochemical and biological properties of phospholipases A(2) from Bothrops atrox snake venom.

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    Kanashiro, Milton M; de Cássia M Escocard, Rita; Petretski, Jorge H; Prates, Maura V; Alves, Elias W; Machado, Olga L T; da Silva, Wilmar Dias; Kipnis, Thereza L

    2002-10-01

    Phospholipases A(2) (PLA(2)s), of molecular mass 13-15kDa, are commonly isolated from snake venom. Two myotoxins with PLA(2) activity, BaPLA(2)I and BaPLA(2)III, with estimated molecular masses of 15kDa were isolated from the venom of Bothrops atrox using Sephacryl S-100-HR and reverse-phase chromatography. BaPLA(2)I was basic, with a pI of 9.1, while BaPLA(2)III was neutral with a pI of 6.9. On a molecular basis, BaPLA(2)III exhibited higher catalytic activity on synthetic substrates than BaPLA(2)I. Comparison of the N-terminal residues of BaPLA(2)I with other PLA(2) proteins from snake venoms showed that it has the highest homology (94%) with B. asper myotoxin II and homology with a PLA(2) Lys(49) from B. atrox (89%). In contrast, BaPLA(2)III demonstrated 75, 72, and 71% homology with PLA(2) from Vipera ammodytes meridionalis, B. jararacussu, and B. jararaca, respectively. BaPLA(2)I and BaPLA(2)III were capable, in vitro, of inducing mast cell degranulation and, in vivo, of causing creatine kinase release, edema, and myonecrosis typical of PLA(2)s from snake venoms, characterized by rapid disruption of the plasma membrane as indicated by clumping of myofilaments and necrosis of affected skeletal muscle cells. BaPLA(2)I- and BaPLA(2)III-specific monoclonal and polyclonal antibodies, although incapable of neutralizing PLA(2) edematogenic activity, blocked myonecrosis efficiently in an in vivo neutralization assay. The results presented herein suggest that the biological active site responsible for edema induction by these two PLA(2) enzymes is distinct from the myonecrosis active site and is not dependent upon the catalytic activity of the PLA(2) enzyme. PMID:12234622

  16. Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox

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    Sousa Marcelo V

    2006-05-01

    Full Text Available Abstract Background Bothrops atrox is responsible for the majority of snakebite accidents in the Brazilian Amazon region. Previous studies have demonstrated that the biological and pharmacological activities of B. atrox venom alter with the age of the animal. Here, we present a comparative proteome analysis of B. atrox venom collected from specimens of three different stages of maturation: juveniles, sub-adults and adults. Results Optimized conditions for two-dimensional gel electrophoresis (2-DE of pooled venom samples were achieved using immobilized pH gradient (IPG gels of non-linear 3–10 pH range during the isoelectric focusing step and 10–20% gradient polyacrylamide gels in the second dimension. Software-assisted analysis of the 2-DE gels images demonstrated differences in the number and intensity of spots in juvenile, sub-adult and adult venoms. Although peptide mass fingerprinting (PMF failed to identify even a minor fraction of spots, it allowed us to group spots that displayed similar peptide maps. The spots were subjected to a combination of tandem mass spectrometry and Mascot and MS BLAST database searches that identified several classes of proteins, including metalloproteinases, serine proteinases, lectins, phospholipases A2, L-amino oxidases, nerve growth factors, vascular endothelial growth factors and cysteine-rich secretory proteins. Conclusion The analysis of B. atrox samples from specimens of different ages by 2-DE and mass spectrometry suggested that venom proteome alters upon ontogenetic development. We identified stage specific and differentially expressed polypeptides that may be responsible for the activities of the venom in each developmental stage. The results provide insight into the molecular basis of the relation between symptomatology of snakebite accidents in humans and the venom composition. Our findings underscore the importance of the use of venoms from individual specimen at various stages of maturation for

  17. Isolation and characterization of a serine protease, Ba III-4, from Peruvian Bothrops atrox venom.

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    Ponce-Soto, L A; Bonfim, V L; Novello, J C; Navarro Oviedo, R; Yarlequé Chocas, A; Marangoni, S

    2007-09-01

    A serine protease from Bothrops atrox (Peruvian specimen's venom) was isolated in two chromatographic steps in LC molecular exclusion and reverse phase-HPLC. This protein was denominated Ba III-4 (33,080.265 Da determinated by MALDI-TOF mass spectrometry) and showed pI of 5.06, Km 0.2 x 10(-1 ) M and the V (máx) 4.1 x 10(-1 )nmoles p-NA/lt/min on the synthetic substrate BapNA. Ba III-4 also showed ability to coagulate bovine fibrinogen. The serine protease was inhibited by soyben trypsin inhibitor and DA2II, which is an anti-hemorrhagic factor isolated from the opossum specie Didelphis albiventris. The primary structure of Ba III-4 showed the presence of His(44), Asp(94) and Ser(193) residues in the corresponding positions to the catalytic triad established in the serine proteases and Ser(193) are inhibited by phenylmethylsulfonylfluoride (PMSF). Amino acid analysis showed a high content of Asp, Glu, Gly, Ser, Ala and Pro, as well as 12 half-cysteine residues. Ba III-4 contained 293 amino acid residues and the primary structure of VIGGDECDIN EHPFLAFMYY SPRYFCGMTL INQEWVLTAA HCRYFCGMTL IHLGVHRESE KANYDEVRRF PKEKYFIFCD NNFTDDEVDK DIMLIRLDKP VSNSEHIAPL SLPSNPPSVG SVCRIMGWGQ TTTSPIDVLS PDEPHCANIN LFDNTVCHTA HPQVANTRTS TDTLCAGDLQ GGRDTCNGDS GGPLICNEQL HGILSWGGDP CAQPNKPAFY TKVYYFDHPW IKSIIAGNKK TVNFTCPPLR SDAKDDSTTY INQEWDWVLT AEHCDRTHMR NSFYDYSSIN SDS. Titration experiments did not show the presence of free sulfhydryl groups after 4 h incubation, nor were differences found in relation to titration kinetics in the presence of nondenaturating buffer. The isolation of this protein, Ba III-4, is of potential interest for the understanding of the pathomechanism of the snake venom action and for the identification of new blood coagulation enzymes of natural sources. PMID:17522968

  18. Bothrops lanceolatus (Fer de lance) venom stimulates leukocyte migration into the peritoneal cavity of mice.

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    Arruda, Vanessa Alves; de Queiroz Guimarães, Alessandra; Hyslop, Stephen; de Araújo, Paulo Maria Ferreira; Bon, Cassian; de Araújo, Albetiza Lôbo

    2003-01-01

    The ability of Bothrops lanceolatus venom to induce neutrophil migration into the peritoneal cavity of mice was investigated. Intraperitoneal injection of venom caused dose- and time-dependent neutrophil migration, which peaked with 750 ng of venom/cavity 4h after venom injection. The neutrophil migration was significantly reduced by pretreatment with dexamethasone (0.5 mg/kg, s.c.), an indirect inhibitor of phospholipase A(2) (PLA(2)), and AA861 (0.01 mg/kg, s.c.), a 5-lipoxygenase inhibitor, but in contrast, was not modified by pretreatment with indomethacin (2 mg/kg, s.c.), an inhibitor of the cyclooxygenase pathway, meloxicam (5 mg/kg, s.c.), an inhibitor of the cyclooxygenase-2 pathway, or the PAF inhibitor WEB2086 (40 mg/kg, s.c.). Dexamethasone and AA861 also inhibited the neutrophil migration by 60% when administered immediately after venom injection, and the coadministration of these two drugs caused a 75% reduction in migration. BLV-induced neutrophil migration was not due to contamination by endotoxin since polymyxin B-treated venom retained its activity. Heating the venom (97 degrees C, 2 min) reduced the PLA(2) activity by 64% and this was accompanied by a corresponding reduction (68%) in neutrophil migration. These results suggest that arachidonate-derived lipoxygenase metabolites (possibly leukotriene B(4)) are involved in the chemotaxis observed. Macrophages may be an important source of these metabolites since the migratory response to venom was potentiated in mice pretreated with thioglycollate, but reduced when the peritoneal cavity was washed with sterile saline. PMID:12467667

  19. Annual changes in seminal variables of golden lanchead pitvipers (Bothrops insularis) maintained in captivity.

    Science.gov (United States)

    Silva, K B; Zogno, M A; Camillo, A B; Pereira, R J G; Almeida-Santos, S M

    2015-12-01

    Bothrops insularis is an endemic and critically endangered snake with an estimated population of 2000 individuals restricted to Queimada Grande Island, in southeastern Brazil. Brazilian researchers established a captive breeding program for the species that includes the application of assisted reproductive technologies. The present study, therefore, aimed to evaluate semen samples from captive B. insularis throughout the year to ascertain seasonal differences in semen traits as well as correlations with body size and weight. Eighteen males with snout-vent length (SVL) ranging from 43.5 to 73.7 cm were collected at quarterly basis between August 2012 and May 2013. Macroscopic analysis revealed semen volumes ranging from 0.5 to 6.0 μL with samples featuring whitish to yellowish color and creamy and thick consistency. Viable sperm was obtained from all males indicating that individuals with SVL equal to or greater than 43.5 cm are sexually developed. However, adult and immature males (estimated by SVL) exhibited different seasonal profiles for motility and progressive motility. Adult males had a decrease in sperm motility and progressive motility during summer and spring, respectively, whereas the same variables did not vary throughout the year in immature snakes. Sperm concentration in all individuals was less (0.5 × 10(9) μL) during the winter, but no seasonal fluctuations were detected in semen volume. These findings are of particular importance to the development of reproductive tools such as male selection, artificial insemination and sperm freezing for the genetic management of this critically endangered snake. PMID:26559333

  20. Inflammatory effects of BaP1 a metalloproteinase isolated from Bothrops asper snake venom: leukocyte recruitment and release of cytokines.

    Science.gov (United States)

    Fernandes, Cristina Maria; Zamuner, Stella Regina; Zuliani, Juliana Pavan; Rucavado, Alexandra; Gutiérrez, José Maria; Teixeira, Catarina de Fátima Pereira

    2006-04-01

    The inflammatory events induced by BaP1, a 22.7 kDa metalloproteinase isolated from Bothrops asper snake venom, were studied. BaP1 i.p. injection in mice induced a marked inflammatory cell infiltrate into peritoneal cavity of animals with predominance of neutrophils in the early phase followed by mononuclear cells in the late period. Inhibition of enzymatic activity of BaP1 by chelation with EDTA resulted in a drastic reduction of this effect. In addition, BaP1 induced a significant increase of blood neutrophil numbers before its accumulation in peritoneal cavity, thus suggesting a stimulatory action of BaP1 on mechanisms of cell mobilization from bone marrow reserve compartments. A reduction in the number of neutrophils was observed in the exudate when antibodies against LECAM-1, CD18 and LFA-1 were used, suggesting the involvement of these adhesion molecules in the effects of BaP1. In contrast, there was no effect with antibodies against ICAM-1 and PECAM-1. Moreover, a conspicuous increment in the levels of IL-1 and TNF-alpha, but not of LTB4, was observed in peritoneal washes collected from mice injected with BaP1. It is concluded that BaP1 induces in vivo a marked leukocyte influx, which parallels an increased number of these cells in the blood, and is associated to the expression of specific leukocyte adhesion molecules and release of chemotactic inflammatory cytokines. Since BaP1 is a P-I class metalloproteinase, these results indicate that the proteolytic domain of metalloproteinases per se can trigger specific inflammatory events. PMID:16529786

  1. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia.

    Science.gov (United States)

    Núñez, V; Otero, R; Barona, J; Saldarriaga, M; Osorio, R G; Fonnegra, R; Jiménez, S L; Díaz, A; Quintana, J C

    2004-07-01

    We determined the neutralizing activity of 12 ethanolic extracts of plants against the edema-forming, defibrinating and coagulant effects of Bothrops asper venom in Swiss Webster mice. The material used consisted of the leaves and branches of Bixa orellana (Bixaceae), Ficus nymphaeifolia (Moraceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plant of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); rhizomes of Renealmia alpinia (Zingiberaceae), Heliconia curtispatha (Heliconiaceae) and Dracontium croatii (Araceae), and the ripe fruit of Citrus limon (Rutaceae). After preincubation of varying amounts of each extract with either 1.0 microg venom for the edema-forming effect or 2.0 microg venom for the defibrinating effect, the mixture was injected subcutaneously (sc) into the right foot pad or intravenously into the tail, respectively, to groups of four mice (18-20 g). All extracts (6.2-200 microg/mouse) partially neutralized the edema-forming activity of venom in a dose-dependent manner (58-76% inhibition), with B. orellana, S. orbicularis, G. panamensis, B. rosademonte, and D. croatii showing the highest effect. Ten extracts (3.9-2000 microg/mouse) also showed 100% neutralizing ability against the defibrinating effect of venom, and nine prolonged the coagulation time induced by the venom. When the extracts were administered either before or after venom injection, the neutralization of the edema-forming effect was lower than 40% for all extracts, and none of them neutralized the defibrinating effect of venom. When they were administered in situ (sc at the same site 5 min after venom injection), the neutralization of edema increased for six extracts, reaching levels up to 64% for C. limon. PMID:15264003

  2. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia.

    Science.gov (United States)

    Núñez, V; Otero, R; Barona, J; Saldarriaga, M; Osorio, R G; Fonnegra, R; Jiménez, S L; Díaz, A; Quintana, J C

    2004-07-01

    We determined the neutralizing activity of 12 ethanolic extracts of plants against the edema-forming, defibrinating and coagulant effects of Bothrops asper venom in Swiss Webster mice. The material used consisted of the leaves and branches of Bixa orellana (Bixaceae), Ficus nymphaeifolia (Moraceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plant of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); rhizomes of Renealmia alpinia (Zingiberaceae), Heliconia curtispatha (Heliconiaceae) and Dracontium croatii (Araceae), and the ripe fruit of Citrus limon (Rutaceae). After preincubation of varying amounts of each extract with either 1.0 microg venom for the edema-forming effect or 2.0 microg venom for the defibrinating effect, the mixture was injected subcutaneously (sc) into the right foot pad or intravenously into the tail, respectively, to groups of four mice (18-20 g). All extracts (6.2-200 microg/mouse) partially neutralized the edema-forming activity of venom in a dose-dependent manner (58-76% inhibition), with B. orellana, S. orbicularis, G. panamensis, B. rosademonte, and D. croatii showing the highest effect. Ten extracts (3.9-2000 microg/mouse) also showed 100% neutralizing ability against the defibrinating effect of venom, and nine prolonged the coagulation time induced by the venom. When the extracts were administered either before or after venom injection, the neutralization of the edema-forming effect was lower than 40% for all extracts, and none of them neutralized the defibrinating effect of venom. When they were administered in situ (sc at the same site 5 min after venom injection), the neutralization of edema increased for six extracts, reaching levels up to 64% for C. limon.

  3. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia

    Directory of Open Access Journals (Sweden)

    Núñez V.

    2004-01-01

    Full Text Available We determined the neutralizing activity of 12 ethanolic extracts of plants against the edema-forming, defibrinating and coagulant effects of Bothrops asper venom in Swiss Webster mice. The material used consisted of the leaves and branches of Bixa orellana (Bixaceae, Ficus nymphaeifolia (Moraceae, Struthanthus orbicularis (Loranthaceae and Gonzalagunia panamensis (Rubiaceae; the stem barks of Brownea rosademonte (Caesalpiniaceae and Tabebuia rosea (Bignoniaceae; the whole plant of Pleopeltis percussa (Polypodiaceae and Trichomanes elegans (Hymenophyllaceae; rhizomes of Renealmia alpinia (Zingiberaceae, Heliconia curtispatha (Heliconiaceae and Dracontium croatii (Araceae, and the ripe fruit of Citrus limon (Rutaceae. After preincubation of varying amounts of each extract with either 1.0 µg venom for the edema-forming effect or 2.0 µg venom for the defibrinating effect, the mixture was injected subcutaneously (sc into the right foot pad or intravenously into the tail, respectively, to groups of four mice (18-20 g. All extracts (6.2-200 µg/mouse partially neutralized the edema-forming activity of venom in a dose-dependent manner (58-76% inhibition, with B. orellana, S. orbicularis, G. panamensis, B. rosademonte, and D. croatii showing the highest effect. Ten extracts (3.9-2000 µg/mouse also showed 100% neutralizing ability against the defibrinating effect of venom, and nine prolonged the coagulation time induced by the venom. When the extracts were administered either before or after venom injection, the neutralization of the edema-forming effect was lower than 40% for all extracts, and none of them neutralized the defibrinating effect of venom. When they were administered in situ (sc at the same site 5 min after venom injection, the neutralization of edema increased for six extracts, reaching levels up to 64% for C. limon.

  4. Immunogenicity of Bothrops atrox (Ophidia: Viperidae venom and its evaluation by immunoenzymatic methods Inmunogenicidad del veneno de Bothrops atrox (Ophidia: Viperidae y su evaluación por métodos inmunoenzimáticos

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    Gustavo A. Sandoval

    2012-02-01

    Full Text Available The immunogenicity of Bothrops atrox, “jergón”, venom was studied using ELISA and Western Blot methods, as well as cross-reactivity patterns against venoms of Bothrops brazili, Lachesis muta and Crotalus durissus. For this purpose, New Zealand white rabbits (2 kg aprox were immunized with four 500 μg doses of B. atrox venom in a period of 90 days. Antibody production was followed using ELISA technique, and title of hiper-immune serum was determined at the end of immunization protocol. Additionally, electrophoretic patterns of venoms were analyzed by SDS-PAGE and venom reactivity against obtained serum by ELISA and Western Blot. Immunization schedule allowed a pronounced antibody production since day 20 of protocol. At the end of process, serum title was 256000, which demonstrated both efficacy and usefulness of the developed procedure. On the other hand, studied venoms showed a heterogenic protein composition according to their electrophoretic patterns, whereas cross-reactivity values of 23,7%, 4,0% and 1,8% were obtained between B. atrox venom and B. brazili, L. muta and C. durissus venoms, respectively, using immunoenzymatic methods. According to our results, this procedure constitutes an initial step for further assays directed to optimization in immunoserum production for envenoming treatment and development of kits for diagnosis and species identification of snakes.Se estudió la inmunogenicidad del veneno de la serpiente Bothrops atrox, “jergón”, utilizando los métodos inmunoenzimáticos de ELISA y Western Blot, así como los patrones de reactividad cruzada empleando los venenos de las serpientes Bothrops brazili, Lachesis muta y Crotalus durissus. Para este fin se inmunizaron conejos albinos Nueva Zelanda (2 kg aprox con cuatro dosis de 500 μg del veneno de B. atrox en un periodo de 90 días. La producción de anticuerpos fue monitoreada mediante la técnica de ELISA, determinándose el título del suero hiperinmune obtenido

  5. Distribution of 131I-labeled Bothrops erythromelas venom in mice

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    C.M.L. Vasconcelos

    1998-03-01

    Full Text Available Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with 131I by the chloramine T method, and separated in a Sephacryl® S-200 column. The efficiency of labeling was 68%. Male Swiss mice (40-45 g, which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc with 0.3 ml (2.35 x 105 cpm/mouse of 131I-venom (N = 42, 131I-albumin or 131I (controls, N = 28 each. Thirty minutes and 1, 3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% NaCl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption in the skin (51% within the first 30 min compared to albumin (20.1% and free iodine (8.2%. Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach (3rd h, small intestine (3rd h and large intestine (6th h. Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively, and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3rd h postinjection (1.98%. There was a low binding of labeled venom in the liver (<0.54%, thyroid (<0.11% and lungs (<0.08%, and no iodinated venom was detected in brain, heart, diaphragm, spleen or bladder. The low venom binding observed in most internal organs, comparable to that of albumin, suggests that B. erythromelas venom does not specifically target most internal organs

  6. Distribution of {sup 131} I- labeled Bothrops erythromelas venom in mice

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    Vasconcelos, C.M.L.; Valenca, R.C.; Araujo, E.A. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia; Modesto, J.C.A.; Pontes, M.M.; Guarnieri, M.C. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Zoologia; Brazil, T.K. [Bahia Univ., Salvador, BA (Brazil). Inst. de Biologia

    1998-03-01

    Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with {sup 131} I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%.Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc) with 0.3 ml (2.35 x 10{sup 5} cpm/mouse) of {sup 131} I-venom (N = 42), {sup 131} -albumin or {sup 131} I (controls, N = 28 each). Thirty minutes and 1,3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% Na Cl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption int he skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach ( 3 rd h),small intestine (3 rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3 rd h postinjection (1.98%).There was a low binding of labeled venom in the liver (B. erythromelas venom does not specifically target most internal organs). That is, the systemic effects of envenomation ar mainly due to an indirect action. (author) 14 refs., 2 figs.

  7. Flora bacteriana da cavidade oral, presas e veneno de Bothrops jararaca: possível fonte de infecção no local da picada Bacterial flora of the oral cavity, fangs and venom of Bothrops jararaca: possible source of infection at the local bite

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    Miguel Tanús Jorge

    1990-02-01

    Full Text Available Materiais colhidos das presas, das bainhas das presas e do veneno de 15 Bothrops jararaca recém-capturadas, aparentemente saudáveis, foram submetidos a exame bacterioscópico e cultura aeróbia a anaeróbia. As bactérias mais freqüentemente isoladas foram os estreptococos do grupo D (1.2 serpentes, Enterobacter sp. (6, Providencia rettgeri (6, Providencia sp. (4, Escherichia coli (4, Morganella morganii (3 e Clostridium sp. (5. Como estas bactérias são semelhantes às encontradas nos abscessos de pacientes picados por serpentes do gênero Bothrops, é válido considerar a possibilidade de que bactérias da boca da serpente sejam inoculadas no momento da picada e, encontrando condições favoráveis de multiplicação, causem infecção.Culture of fang, fang sheath and venom of fifteen healthy freshly captured Bothrops jararaca were analised. The bacteria most frequently encountered were group D streptococci (12 snakes, Enterobacter sp. (6, Providencia rettgeri (6, Providencia sp. (4, Escherichia coli (4, Morganella morganii (3 and Clostridium sp. (5. The bacteria observed are similar to those found in the abscesses from Bothrops bitten patients. Since these snake mouth bacteria may be inoculated during the snake bite, bacterial multiplication and infection may occur under favorable conditions.

  8. Another new and threatened species of lancehead genus Bothrops (Serpentes, Viperidae) from Ilha dos Franceses, Southeastern Brazil.

    Science.gov (United States)

    Barbo, Fausto E; Gasparini, João Luiz; Almeida, Antonio P; Zaher, Hussam; Grazziotin, Felipe G; Gusmão, Rodrigo B; Ferrarini, José Mário G; Sawaya, Ricardo J

    2016-01-01

    A new insular species of the genus Bothrops is described from Ilha dos Franceses, a small island off the coast of Espírito Santo State, in southeastern Brazil. The new species differs from mainland populations of B. jararaca mainly by its small size, relative longer tail, relative smaller head length, and relative larger eyes. The new species is distinguished from B. alcatraz, B. insularis and B. otavioi by the higher number of ventral and subcaudal scales, relative longer tail and smaller head. The new species is highly abundant on the island, being nocturnal, semiarboreal, and feeding on small lizards and centipeds. Due its unique and restricted area of occurrence, declining quality of habitat, and constant use of the island for tourism, the new species may be considered as critically endangered. PMID:27394563

  9. Isolation and characterization of a myotoxin from Bothrops brazili Hoge, 1953 Hoge, 1953 snake venom (Ophidia: Viperidae.

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    Carmen Pantigoso

    2014-06-01

    Full Text Available A myotoxin from the venom of the snake Bothrops brazili has been purified by ion-exchange chromatography on CM-Sephadex C-50 with 0,05 M ammonium acetate buffer pH 7. The homogeneity was evaluated by PAGE with and without SDS, immunodiffusion and immunoelectrophoresis. The myotoxin is a basic protein with 15,6% of Lys+Arg; it is not a glicoprotein, has not enzymatic activity, and corresponds to 25% of the whole venom protein. The molecular weight of the myotoxin was determined by PAGE-SDS and gel filtration chromatography. The myotoxin has 30 KDa of molecular weight and two polypeptide chains of 15 KDa each. Myotoxin produces a severe necrosis on the gastrocnemius muscle of white mice. The myotoxin does not have hemolytic nor anticoagulant activity. However, produces edema with a DEM of 32,6 mg of protein.

  10. Description of Serpentirhabdias atroxi n. sp. (Nematoda: Rhabdiasidae), a parasite of Bothrops atrox (Linnaeus) (Reptilia: Serpentes: Viperidae) in Brazilian Amazonia.

    Science.gov (United States)

    Kuzmin, Yuriy; Giese, Elane Guerreiro; Melo, Francisco Tiago de Vasconcelos; da Costa, Paulo André Ferreira Borges; Maschio, Gleomar Fabiano; dos Santos, Jeannie Nascimento

    2016-01-01

    A new lung-dwelling nematode species is described from the common lancehead Bothrops atrox (Linnaeus) in the Brazilian Amazon Region. The species is assigned to the genus Serpentirhabdias Tkach, Kuzmin & Snyder, 2014 based on the presence of six lips arranged in two lateral groups, the absence of prominent cuticular inflations, and lung parasitism in snakes. Serpentirhabdias atroxi n. sp. differs from other species of the genus mainly by details of the morphology of the anterior end: cuticularised ring surrounding the anterior part of the buccal cavity and six minute onchia present in the oesophastome. Serpentirhabdias atroxi n. sp. is the seventh species of the genus known from the Neotropical Realm and the second species described from viperid snakes.

  11. Purification and Characterization of Jararassin-I,A Thrombin-like Enzyme from Bothrops jararaca Snake Venom

    Institute of Scientific and Technical Information of China (English)

    Débora F. VIEIRA; Leandra WATANABE; Carolina D. SANT'ANA; Silvana MARCUSSI; Suely V. SAMPAIO; Andreimar M. SOARES; Raghuvir K. ARNI

    2004-01-01

    A thrombin-like serine protease, jararassin-I, was isolated from the venom of Bothrops jararaca. The protein was obtained in high yield and purity by a single chromatographic step using the affinity resin Benzamidine-Sepharose CL-6B. SDS-PAGE and dynamic light scattering analyses indicated that the molecular mass of the enzyme was about 30 kD. The enzyme possessed fibrinogenolytic and coagulant activities. The jararassin-I degraded the Bβ chain of fibrinogen while the Aα chain and γ chain were unchanged.Proteases inhibitors, PMSF and benzamidine inhibited the coagulant activity. These results showed jararassinI is a serine protease similar to coagulating thrombin-like snake venom proteases, but it specifically cleaves Bβ chain of bovine fibrinogen. Single crystals of enzyme were obtained (0.2 mm×0.2 mm×0.2 mm) and used for X-ray diffraction experiments.

  12. Aislamiento y caracterización parcial de una miotoxina del veneno de la serpiente Bothrops atrox (Ophidia: Viperidae

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    Sergio Huatuco

    2013-06-01

    Full Text Available Se ha purificado una miotoxina del veneno de la serpiente Bothrops atrox, empleando una columna de intercambio catiónico de CM-Sephadex C-50 equilibrada con buffer acetato de amonio 0,05M a pH 7. La miotoxina es una proteína básica, y por cromatografía de filtración y PAGE-SDS se ha determinado que tiene un peso molecular de 27 kDa, estando formada por dos cadenas polipeptídicas de 14 kDa cada una. La inoculación de la miotoxina en el músculo gastrocnemius de ratones albinos produce la liberación de creatina kinasa así como la necrosis del tejido. La miotoxina tiene actividad de fosfolipasa, anticoagulante y edemática, pero no hemolítica.

  13. Description of Serpentirhabdias atroxi n. sp. (Nematoda: Rhabdiasidae), a parasite of Bothrops atrox (Linnaeus) (Reptilia: Serpentes: Viperidae) in Brazilian Amazonia.

    Science.gov (United States)

    Kuzmin, Yuriy; Giese, Elane Guerreiro; Melo, Francisco Tiago de Vasconcelos; da Costa, Paulo André Ferreira Borges; Maschio, Gleomar Fabiano; dos Santos, Jeannie Nascimento

    2016-01-01

    A new lung-dwelling nematode species is described from the common lancehead Bothrops atrox (Linnaeus) in the Brazilian Amazon Region. The species is assigned to the genus Serpentirhabdias Tkach, Kuzmin & Snyder, 2014 based on the presence of six lips arranged in two lateral groups, the absence of prominent cuticular inflations, and lung parasitism in snakes. Serpentirhabdias atroxi n. sp. differs from other species of the genus mainly by details of the morphology of the anterior end: cuticularised ring surrounding the anterior part of the buccal cavity and six minute onchia present in the oesophastome. Serpentirhabdias atroxi n. sp. is the seventh species of the genus known from the Neotropical Realm and the second species described from viperid snakes. PMID:26739285

  14. Crystallization and preliminary crystallographic studies of a phospholipase A2 from the venom of the Brazilian snake Bothrops moojeni.

    Science.gov (United States)

    Nonato, M C; Garratt, R C; Mascarenhas, Y P; Jesus, W D; Assakura, M T; Serrano, S M; Oliva, G

    2001-04-01

    A phospholipase A(2) purified from the venom of the snake Bothrops moojeni has been crystallized by vapour-diffusion techniques in hanging drops at 291 K. The crystals, which were grown in the absence of Ca(2+), belong to the cubic system, space group P432, with unit-cell parameters a = b = c = 91.86 A, and contain one molecule in the asymmetric unit (V(M) = 2.71 A(3) Da(-1)). X-ray diffraction experiments provide data to 2.35 A resolution collected on a rotating-anode home source at cryogenic temperatures. The structure has been solved via molecular-replacement techniques using a single monomer of the crystallographic structure of the phospholipase from the Western diamondback rattlesnake (Crotalus atrox) as a search model. PMID:11264594

  15. Preliminary assessment of Hedychium coronarium essential oil on fibrinogenolytic and coagulant activity induced by Bothrops and Lachesis snake venoms

    Science.gov (United States)

    2014-01-01

    Background The search for new inhibitors of snake venom toxins is essential to complement or even replace traditional antivenom therapy, especially in relation to compounds that neutralize the local effects of envenomations. Besides their possible use as alternative to traditional antivenom therapy, some plant species possess bioactive secondary metabolites including essential oils, which can be extracted from weeds that are considered substantial problems for agriculture, such as Hedychium coronarium. Methods The essential oils of leaves and rhizomes from H. coronarium were extracted by hydrodistillation, and their potential inhibitory effects on the coagulant and fibrinogenolytic activities induced by the venoms of Lachesis muta, Bothrops atrox and Bothrops moojeni were analyzed. Citrated human plasma was used to evaluate the clotting time whereas changes in fibrinogen molecules were visualized by electrophoresis in polyacrylamide gel. The experimental design used for testing coagulation inhibition was randomized in a 3 × 2 factorial arrangement (concentration × essential oils), with three replications. The essential oils were compared since they were extracted from different organs of the same botanical species, H. coronarium. Results The results suggest that the oils interact with venom proteases and plasma constituents, since all oils evaluated, when previously incubated with venoms, were able to inhibit the clotting effect, with less inhibition when oils and plasma were preincubated prior to the addition of venoms. Conclusions Thus, after extensive characterization of their pharmacological and toxicological effects, the essential oils can be used as an alternative to complement serum therapy, especially considering that these plant metabolites generally do not require specific formulations and may be used topically immediately after extraction. PMID:26413083

  16. Aqueous Leaf Extract of Jatropha gossypiifolia L. (Euphorbiaceae) Inhibits Enzymatic and Biological Actions of Bothrops jararaca Snake Venom

    Science.gov (United States)

    Félix-Silva, Juliana; Souza, Thiago; Menezes, Yamara A. S.; Cabral, Bárbara; Câmara, Rafael B. G.; Silva-Junior, Arnóbio A.; Rocha, Hugo A. O.; Rebecchi, Ivanise M. M.; Zucolotto, Silvana M.; Fernandes-Pedrosa, Matheus F.

    2014-01-01

    Snakebites are a serious public health problem due their high morbi-mortality. The main available specific treatment is the antivenom serum therapy, which has some disadvantages, such as poor neutralization of local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies is relevant. Therefore, the aim of this study was to evaluate the antiophidic properties of Jatropha gossypiifolia, a medicinal plant used in folk medicine to treat snakebites. The aqueous leaf extract of the plant was prepared by decoction and phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins. The extract was able to inhibit enzymatic and biologic activities induced by Bothrops jararaca snake venom in vitro and in vivo. The blood incoagulability was efficiently inhibited by the extract by oral route. The hemorrhagic and edematogenic local effects were also inhibited, the former by up to 56% and the latter by 100%, in animals treated with extract by oral and intraperitoneal routes, respectively. The inhibition of myotoxic action of B. jararaca reached almost 100%. According to enzymatic tests performed, it is possible to suggest that the antiophidic activity may be due an inhibitory action upon snake venom metalloproteinases (SVMPs) and/or serine proteinases (SVSPs), including fibrinogenolytic enzymes, clotting factors activators and thrombin like enzymes (SVTLEs), as well upon catalytically inactive phospholipases A2 (Lys49 PLA2). Anti-inflammatory activity, at least partially, could also be related to the inhibition of local effects. Additionally, protein precipitating and antioxidant activities may also be important features contributing to the activity presented. In conclusion, the results demonstrate the potential antiophidic activity of J. gossypiifolia extract, including its significant action upon local effects, suggesting that

  17. Aqueous leaf extract of Jatropha gossypiifolia L. (Euphorbiaceae inhibits enzymatic and biological actions of Bothrops jararaca snake venom.

    Directory of Open Access Journals (Sweden)

    Juliana Félix-Silva

    Full Text Available Snakebites are a serious public health problem due their high morbi-mortality. The main available specific treatment is the antivenom serum therapy, which has some disadvantages, such as poor neutralization of local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies is relevant. Therefore, the aim of this study was to evaluate the antiophidic properties of Jatropha gossypiifolia, a medicinal plant used in folk medicine to treat snakebites. The aqueous leaf extract of the plant was prepared by decoction and phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins. The extract was able to inhibit enzymatic and biologic activities induced by Bothrops jararaca snake venom in vitro and in vivo. The blood incoagulability was efficiently inhibited by the extract by oral route. The hemorrhagic and edematogenic local effects were also inhibited, the former by up to 56% and the latter by 100%, in animals treated with extract by oral and intraperitoneal routes, respectively. The inhibition of myotoxic action of B. jararaca reached almost 100%. According to enzymatic tests performed, it is possible to suggest that the antiophidic activity may be due an inhibitory action upon snake venom metalloproteinases (SVMPs and/or serine proteinases (SVSPs, including fibrinogenolytic enzymes, clotting factors activators and thrombin like enzymes (SVTLEs, as well upon catalytically inactive phospholipases A2 (Lys49 PLA2. Anti-inflammatory activity, at least partially, could also be related to the inhibition of local effects. Additionally, protein precipitating and antioxidant activities may also be important features contributing to the activity presented. In conclusion, the results demonstrate the potential antiophidic activity of J. gossypiifolia extract, including its significant action upon local effects

  18. Bothrops asper snake venom and its metalloproteinase BaP–1 activate the complement system. Role in leucocyte recruitment

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    Sandra H. P. Farsky

    2000-01-01

    Full Text Available The venom of the snake Bothrops asper, the most important poisonous snake in Central America, evokes an inflammatory response, the mechanisms of which are not well characterized. The objectives of this study were to investigate whether B. asper venom and its purified toxins – phospholipases and metalloproteinase – activate the complement system and the contribution of the effect on leucocyte recruitment. In vitro chemotaxis assays were performed using Boyden's chamber model to investigate the ability of serum incubated with venom and its purified toxins to induce neutrophil migration. The complement consumption by the venom was evaluated using an in vitro haemolytic assay. The importance of complement activation by the venom on neutrophil migration was investigated in vivo by injecting the venom into the peritoneal cavity of C5-deficient mice. Data obtained demonstrated that serum incubated with crude venom and its purified metalloproteinase BaP–1 are able to induce rat neutrophil chemotaxis, probably mediated by agent(s derived from the complement system. This hypothesis was corroborated by the capacity of the venom to activate this system in vitro. The involvement of C5a in neutrophil chemotaxis induced by venom-activated serum was demonstrated by abolishing migration when neutrophils were pre-incubated with antirat C5a receptor antibody. The relevance of the complement system in in vivo leucocyte mobilization was further demonstrated by the drastic decrease of this response in C5-deficient mice. Pre-incubation of serum with the soluble human recombinant complement receptor type 1 (sCR 1 did not prevent the response induced by the venom, but abolished the migration evoked by metalloproteinase-activated serum. These data show the role of the complement system in bothropic envenomation and the participation of metalloproteinase in the effect. Also, they suggest that the venom may contain other component(s which can cause direct activation

  19. Biochemical Characterization, Action on Macrophages, and Superoxide Anion Production of Four Basic Phospholipases A2 from Panamanian Bothrops asper Snake Venom

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    Aristides Quintero Rueda

    2013-01-01

    Full Text Available Bothrops asper (Squamata: Viperidae is the most important venomous snake in Central America, being responsible for the majority of snakebite accidents. Four basic PLA2s (pMTX-I to -IV were purified from crude venom by a single-step chromatography using a CM-Sepharose ion-exchange column (1.5 × 15 cm. Analysis of the N-terminal sequence demonstrated that pMTX-I and III belong to the catalytically active Asp49 phospholipase A2 subclass, whereas pMTX-II and IV belong to the enzymatically inactive Lys49 PLA2s-like subclass. The PLA2s isolated from Panama Bothrops asper venom (pMTX-I, II, III, and IV are able to induce myotoxic activity, inflammatory reaction mainly leukocyte migration to the muscle, and induce J774A.1 macrophages activation to start phagocytic activity and superoxide production.

  20. ESI-MS/MS identification of a bradykinin-potentiating peptide from Amazon Bothrops atrox snake venom using a hybrid Qq-oaTOF mass spectrometer.

    Science.gov (United States)

    Coutinho-Neto, Antonio; Caldeira, Cleópatra A S; Souza, Gustavo H M F; Zaqueo, Kayena D; Kayano, Anderson M; Silva, Rodrigo S; Zuliani, Juliana P; Soares, Andreimar M; Stábeli, Rodrigo G; Calderon, Leonardo A

    2013-02-01

    A bradykinin-potentiating peptide (BPP) from Amazon Bothrops atrox venom with m/z 1384.7386 was identified and characterized by collision induced dissociation (CID) using an ESI-MS/MS spectra obtained in positive ion mode on a hybrid Qq-oaTOF mass spectrometer, Xevo G2 QTof MS (Waters, Manchester, UK). De novo peptide sequence analysis of the CID fragmentation spectra showed the amino acid sequence ZKWPRPGPEIPP, with a pyroglutamic acid and theoretical monoisotopic m/z 1384.7378, which is similar to experimental data, showing a mass accuracy of 0.6 ppm. The peptide is homologous to other BPP from Bothrops moojeni and was named as BPP-BAX12. PMID:23430539

  1. ESI-MS/MS Identification of a Bradykinin-Potentiating Peptide from Amazon Bothrops atrox Snake Venom Using a Hybrid Qq-oaTOF Mass Spectrometer

    Directory of Open Access Journals (Sweden)

    Leonardo A. Calderon

    2013-02-01

    Full Text Available A bradykinin-potentiating peptide (BPP from Amazon Bothrops atrox venom with m/z 1384.7386 was identified and characterized by collision induced dissociation (CID using an ESI-MS/MS spectra obtained in positive ion mode on a hybrid Qq-oaTOF mass spectrometer, Xevo G2 QTof MS (Waters, Manchester, UK. De novo peptide sequence analysis of the CID fragmentation spectra showed the amino acid sequence ZKWPRPGPEIPP, with a pyroglutamic acid and theoretical monoisotopic m/z 1384.7378, which is similar to experimental data, showing a mass accuracy of 0.6 ppm. The peptide is homologous to other BPP from Bothrops moojeni and was named as BPP-BAX12.

  2. An Isoflavone from Dipteryx alata Vogel is Active against the in Vitro Neuromuscular Paralysis of Bothrops jararacussu Snake Venom and Bothropstoxin I, and Prevents Venom-Induced Myonecrosis

    OpenAIRE

    Miriéle C. Ferraz; Edson H. Yoshida; Renata V.S. Tavares; Cogo, José C; Adélia C.O. Cintra; Dal Belo, Cháriston A.; Franco, Luiz M.; dos Santos, Márcio G; Flávia A Resende; Eliana A. Varanda; Stephen Hyslop; Pilar Puebla; Arturo San Feliciano; Yoko Oshima-Franco

    2014-01-01

    Snakebite is a neglected disease and serious health problem in Brazil, with most bites being caused by snakes of the genus Bothrops. Although serum therapy is the primary treatment for systemic envenomation, it is generally ineffective in neutralizing the local effects of these venoms. In this work, we examined the ability of 7,8,3'-trihydroxy-4'-methoxyisoflavone (TM), an isoflavone from Dipteryx alata, to neutralize the neurotoxicity (in mouse phrenic nerve-diaphragm preparations) and myoto...

  3. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia

    OpenAIRE

    Núñez V.; Otero R.; Barona J.; Saldarriaga M.; Osorio R.G.; Fonnegra R.; Jiménez S.L.; Díaz A.; Quintana J.C.

    2004-01-01

    We determined the neutralizing activity of 12 ethanolic extracts of plants against the edema-forming, defibrinating and coagulant effects of Bothrops asper venom in Swiss Webster mice. The material used consisted of the leaves and branches of Bixa orellana (Bixaceae), Ficus nymphaeifolia (Moraceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plant of Pleop...

  4. Sexual dimorphism in development and venom production of the insular threatened pit viper Bothrops insularism (Serpentes: Viperidae) of Queimada Grande Island, Brazil

    OpenAIRE

    S.R. Travaglia-Cardoso; A. Zelanis; M. F. D. Furtado

    2010-01-01

    Bothrops insularis is a threatened snake endemic to Queimada Grande Island, southern coast of São Paulo, Brazil, and the occurrence of sexual abnormalities in females (females with functional ovaries and rudimentary hemipenis) has been reported in this population. To date there are few data regarding developmental features of this particular species. The aim of this study was to follow some developmental features in specimens maintained in captivity for seven years in the Herpetology Labora...

  5. Increments in cytokines and matrix metalloproteinases in skeletal muscle after injection of tissue-damaging toxins from the venom of the snake Bothrops asper

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    Alexandra Rucavado

    2002-01-01

    Full Text Available Envenomations by the snake Bothrops asper are characterized by prominent local tissue damage (i.e. myonecrosis, blistering, hemorrhage and edema. Various phospholipases A2 and metalloproteinases that induce local pathological alterations have been purified from this venom. Since these toxins induce a conspicuous inflammatory response, it has been hypothesized that inflammatory mediators may contribute to the local pathological alterations described. This study evaluated the local production of cytokines and matrix metalloproteinases (MMPs as a consequence of intramuscular injections of an Asp-49 myotoxic phospholipase A2 (myotoxin III (MT-III and a P-I type hemorrhagic metalloproteinase (BaP1 isolated from B. asper venom. Both enzymes induced prominent tissue alterations and conspicuous increments in interleukin (IL-1β, IL-6 and a number of MMPs, especially gelatinase MMP-9, rapidly after injection. In contrast, no increments in tumor necrosis factor-α (TNF-α and interferon-γ were detected. In agreement, MT-III and BaP1 did not induce the synthesis of TNF-α by resident peritoneal macrophages in vitro. Despite the conspicuous expression of latent forms of MMPs in muscle, evidenced by zymography, there were no increments in activated MMP-2 and only a small increase in activated MMP-9, as detected by a functional enzymatic assay. This suggests that MMP activity was regulated by a highly controlled activation of latent forms and, probably, by a concomitant synthesis of MMP inhibitors. Since no hemorrhage nor dermonecrosis were observed after injection of MT-III, despite a prominent increase in MMP expression, and since inflammatory exudate did not enhance hemorrhage induced by BaP1, it is suggested that endogenous MMPs released in the tissue are not responsible for the dermonecrosis and hemorrhage characteristic of B. asper envenomation. Moreover, pretreatment of mice with the peptidomimetic MMP inhibitor batimastat did not reduce myotoxic nor

  6. Análisis inmunoenzimático (ELISA para determinar niveles de IgG anti Bothrops atrox en accidente ofídico Immunoenzymatic determination of IgG anti bothrops atrox levels after snake bites

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    John J. Estrada

    1993-01-01

    Full Text Available Se desarrolló un método de inmunización de conejos con veneno de Bothrops atrox con el fin de preparar antisueros y estandarizar un inmunoanálisis (ELISA para medir niveles de IgG en pacientes con accidente ofídico. La respuesta Inmune de los conejos se siguió por inmunodifusión en doble dimensión (Ouchterlony e inmunoelectroforesis, demostrando la presencia de bandas nítidas desde el día 60 y en todas las sangrias posteriores; se comprobó que hay variabilidad individual en su respuesta inmune. El ELISA para detección de IgG humana anti B. atrox en los indígenas del Chocó fue una prueba simple y sensible (83.3% pero inespecífica por las reacciones cruzadas en individuos que habían sufrido accidentes por B. nasutus. La técnica para detectar IgG equina anti B. atrox en pacientes tratados con antiveneno fue tambIén simple y muy sensible. We developed an immunization method for the production of rabbit antisera against Bothrops atroxvenoms. An enzyme-Ilnked assay (ELISA was standardized in order to measure IgG levels after snake bites. The immune response of rabbits, as determined by Ouchterlony and immunoelectrophoresis techniques, revealed bands of precipitation from the sixtieth day on. Individual variability in the immune response of rabbits was demonstrated. For the measurement of IgG levels In Indians from the Department of Choco (Colombia, ELISA proved to be a sensitive (83.3% and simple but not an specific procedure, since there were cross-reactions in those previously bitten by B. nasutus. ELISA was also simple and sensitive (100% for the determination of equine anti B. atrox IgG antibodies in patients treated with antivenom

  7. Description of a new ascaridoid parasite of Bothrops jararaca Wied (Reptilia, Ophidia in Brazil Descrição de um novo ascarídeo parasito de Bothrops jararaca Wied (Reptilia, Ophidia no Brasil

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    Liege R. Siqueira

    2005-09-01

    Full Text Available A new species of Ophidascaris Baylis, 1920 is described on the basis of specimens recovered from the pit viper Bothrops jararaca Wied, 1824 in Brazil. The new species referred here was compared to other of the genus that are the closest. The differentiation was based on reliable morphological characters, mainly on what refers to the spicular length, number and distribution of caudal papillae, location of the vulvar opening, eggs size and aspect of the labial indentations. Moreover, female specimens are the only to possess a most prominent and characteristic post-anal tubercle-like structure and when analyzed together with the males, that show a great number of pre-cloacal papillae, the presence of an unpaired papilla and also of a less conspicuous post-cloacal tubercle-like structure, permit the prompt identification of the new species.Uma nova espécie de Ophidascaris Baylis, 1920 é descrita, com base em exemplares parasitos de jararaca, Bothrops jararaca Wied, 1824 no Brasil. A nova espécie agora descrita foi comparada a outras do gênero que lhe são as mais próximas. A diferenciação foi baseada em caracteres morfológicos consistentes, principalmente nos que dizem respeito ao tamanho dos espículos, número e distribuição de papilas caudais, localização da abertura vulvar, tamanho dos ovos e aspecto das indentações labiais. Além disso, os exemplares fêmeas são os únicos no gênero a apresentarem uma estrutura em forma de tubérculo pós-anal bastante característica e proeminente, quando analisados junto aos exemplares machos, nos quais existe um grande número de papilas pré-cloacais, presença de papila caudal ímpar e, também, da estrutura pós-cloacal em forma de tubérculo, embora menos evidente que nas fêmeas, permitem a pronta identificação da espécie nova.

  8. Bothrops jararaca venom (BjV) induces differential leukocyte accumulation in mice genetically selected for acute inflammatory reaction: the role of host genetic background on expression of adhesion molecules and release of endogenous mediators.

    Science.gov (United States)

    Carneiro, Adriana S; Ribeiro, Orlando G; Cabrera, Wafa H K; Vorraro, Francisca; De Franco, Marcelo; Ibañez, Olga M; Starobinas, Nancy

    2008-10-01

    The dynamics of the local inflammatory events induced by Bothrops jararaca venom (BjV) inoculation in footpad of mice genetically selected for maximal (AIRmax) and minimal (AIRmin) acute inflammatory reactivity (AIR) was investigated. The BjV injection induced a marked inflammatory cell infiltrate with predominance of neutrophils, with increased blood cell numbers before its accumulation, suggesting a stimulatory action of BjV on mechanisms of cell mobilization from bone marrow. The process of cell migration is regulated by different cell-adhesion molecules (CAM). Our results showed that neutrophil cells from both lines had the same pattern of response concerning CAMs expression, presenting the involvement of l-selectin, Mac-1 and PECAM-1 adhesion molecules in BjV-induced neutrophil accumulation. The effect of BjV on the release of pro-inflammatory cytokines and chemokines related with cellular migration was also studied and IL-1beta, IL-6, TNF-alpha and MIP-2 levels could be detected after venom injection. The AIRmax mice were shown to be more responsive than AIRmin with respect to leukocyte influx, expression of MIP-2 and release of IL-1beta and IL-6. These results demonstrate the importance of host genetic background in the local response and the involvement of alleles accumulated in AIRmax mice in the inflammatory events induced by BjV. PMID:18723041

  9. Linear B-cell epitopes in BthTX-1, BthTX-II and BthA-1, phospholipase A₂'s from Bothrops jararacussu snake venom, recognized by therapeutically neutralizing commercial horse antivenom.

    Science.gov (United States)

    De-Simone, Salvatore G; Napoleão-Pego, Paloma; Teixeira-Pinto, Luiz A L; Santos, Jonathas D L; De-Simone, Thatiane S; Melgarejo, Anibal R; Aguiar, Aniesse S; Marchi-Salvador, Daniela P

    2013-09-01

    The benefits from treatment with antivenom sera are indubitable. However, the mechanism for toxin neutralization has not been completely elucidated. A mixture of anti-bothropic and anti-crotalic horse antivenom has been reported to be more effective in neutralizing the effects of Bothrops jararacussu snake venom than anti-bothropic antivenom alone. This study determined which regions in the three PLA₂s from B. jararacussu snake venom are bound by antibodies in tetravalent anti-bothropic and monovalent anti-crotalic commercial horse antivenom. Mapping experiments of BthTX-I, BthTX-II and BthA-I using two small libraries of 69 peptides each revealed six major IgG-binding epitopes that were recognized by both anti-bothropic and anti-crotalic horse antivenom. Two epitopes in BthTX-I were only recognized by the anti-bothropic horse antivenom, while anti-crotalic horse antivenom recognized four unique epitopes across the three PLA₂s. Our studies suggest that the harmful activities of the PLA₂s present in the venom of B. jararacussu are neutralized by the combinatorial treatment with both antivenom sera through their complementary binding sites, which provides a wide coverage on the PLA₂s. This is the first peptide microarray of PLA₂s from B. jararacussu snake venom to survey the performance of commercial horse antiophidic antivenom. Regions recognized by the protective antivenom sera are prime candidates for improved venom cocktails or a chimeric protein encoding the multiple epitopes to immunize animals as well as for designing future synthetic vaccines. PMID:23792452

  10. Crystal structure of a Bothrops toxin I, a Myotoxin K 49 like Phospholipase A{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Giotto, M.T.S.; Azevedo, W.F.; Horjales, E.; Oliva, G.; Mascarenhas, Y.P.; Garratt, R.C. [Sao Paulo Univ., Sao Carlos, SP (Brazil). Inst. de Fisica]|[Sao Paulo Univ., Ribeirao Preto, SP (Brazil). Faculdade de Medicina

    1996-12-31

    Full text. Bothrops toxin-I (BthTX-I) is a mytoxin isolated from the Brazilian snake Bothrops jararacussu which is a member of the phospholipase A{sub 2} but presents no catalytic activity due to a D49K substitution. Protein was provided by Prof. Dr. J.R. Giglio and Prof{sup a}. Dr{sup a}. A.C.O. Cintra from the Departamento de Medicina de Ribeirao Preto and used in crystallization experiments which were performed using the vapor diffusion technique in hanging drops at 18{sup 0}C. The BthTX-I crystallized in 0.1 M HEPES, pH ranging from 7.0 to 7.6. The precipitant was (NH{sub 4})SO{sub 4} in concentrations ranging from 7.0 to 7.6. The precipitant was (NH{sub 4})SO{sub 4} in concentrations ranging from 57 to 62%. The data collection was initially performed using the automatic difractometer R-AXIS IIC from the Rigaku Co. at the Laboratorio de Cristalografia of IFESC-USP. Subsequently a second data set was collected at SERC Daresbury Laboratory in England using synchrotron radiation. BthTX-I crystallizes in space group P3{sub 1} 21 with a=b=57.58 A, c=131.27 A, {alpha}={beta}=900{sup 0}C and {gamma}=120{sup 0}C. Processing of the data was performed with the MOSFLM program yielding an R{sub merge}= 6.3% and completeness of 99.6% at 2.1 A of resolution. The structure was solved by Molecular Replacement using the package AMoRe with the Agksitrodon piscivorus piscivorus enyme as search model and refined using the refinement program X-PLOR to a final R=18.7% and R{sub free}=27.4%. The assymetric unit contains two monomers, which can be chosen such that they present similar interactions to those described for the homogeneous myotoxin II from B. asper. The interface is, however, surprisingly small when compared to other dimeric structures and is less complementary than expected. A theoretical model for phospholipid building to BthTX-I suggests that no direct interactions between the sn-2 ester bond and K49 would be expected, thus explaining the lack of catalytic activity.

  11. The crucial role of the MyD88 adaptor protein in the inflammatory response induced by Bothrops atrox venom.

    Science.gov (United States)

    Moreira, Vanessa; Teixeira, Catarina; Borges da Silva, Henrique; D'Império Lima, Maria Regina; Dos-Santos, Maria Cristina

    2013-06-01

    Most snake accidents in North Brazil are attributed to Bothrops atrox, a snake species of the Viperidae family whose venom simultaneously induces local and systemic effects in the victims. The former are clinically more important than the latter, as they cause severe tissue lesions associated with strong inflammatory responses. Although several studies have shown that inflammatory mediators are produced in response to B. atrox venom (BaV), there is little information concerning the molecular pathways involved in innate immune system signaling. Myeloid differentiation factor 88 (MyD88) is an adaptor molecule responsible for transmitting intracellular signals from most toll-like receptors (TLRs) after they interact with pathogen-associated molecular patterns (PAMPs) or other stimuli such as endogenous damage-associated molecular patterns (DAMPs). The MyD88-dependent pathway leads to activation of transcription factors, which in turn induce synthesis of inflammatory mediators such as eicosanoids, cytokines and chemokines. The aim of this study was to investigate the involvement of MyD88 on the acute inflammatory response induced by BaV. Wild-type (WT) C57BL/6 mice and MyD88 knockout (MyD88(-/-)) mice were intraperitoneally injected with BaV. Compared to WT mice, MyD88(-/-) animals showed an impaired inflammatory response to BaV, with lower influx of polymorphonuclear and mononuclear cells to the peritoneal cavity. Furthermore, peritoneal leukocytes from BaV-injected MyD88(-/-) mice did not induce COX-2 or LTB4 protein expression and released low concentrations of PGE2. These mice also failed to produce Th1 and Th17 cytokines and CCL-2, but IL-10 levels were similar to those of BaV-injected WT mice. Our results indicate that MyD88 signaling is required for activation of the inflammatory response elicited by BaV, raising the possibility of developing new therapeutic targets to treat Bothrops sp. poisoning. PMID:23474268

  12. Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    A.V Pérez

    2008-01-01

    Full Text Available A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 µg and fibrinogen (minimum coagulant dose = 4.2 µg in vitro, and promotes defibrin(ogenation in vivo (minimum defibrin(ogenating dose = 1.0 µg. In addition, when injected intravenously in mice at doses of 5 and 10 µg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the `gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.

  13. Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper

    International Nuclear Information System (INIS)

    The venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investigate the possible presence of inhibitors of platelet aggregation. Using a combination of gel filtration, anion-exchange chromatography, and reverse-phase high performance liquid chromatography, we isolated an acidic protein which shows a single chain composition, with a molecular mass of ∼8 kDa, estimated by SDS-polyacrylamide gel electrophoresis. Its N-terminal sequence has high similarity to disintegrins isolated from different snake venoms, which are known to bind to cellular integrins such as the GPIIb/IIIa fibrinogen receptor on platelets. The purified protein exerted potent aggregation inhibitory activity on ADP-stimulated human platelets in vitro, with an estimated IC50 of 50 nM. This biological activity, together with the biochemical characteristics observed, demonstrate that the protein isolated from B. asper venom is a disintegrin, hereby named bothrasperin. This is the first disintegrin isolated from Central American viperid snake species. (Author)

  14. Biology and conservation status of Piraja’s Lancehead Snake Bothrops piraña Amaral, 1923 (Serpentes: Viperidae, Brazil

    Directory of Open Access Journals (Sweden)

    M.A.D. Freitas

    2014-09-01

    Full Text Available The venomous snake Bothrops pirajai (Amaral, 1923 is endemic to Brazil. Despite being described almost a century ago, very little is known about this species, which has never been studied in situ. Here, we present new data on the biology and natural history of B. pirajai based on a review of existing museum specimens and a field study (1504 man hours carried out throughout the species range. The distribution of B. pirajai was found to be restricted to the eastern Atlantic forest of the state of Bahia, Brazil, between Todosos Santos Bay, Itabuna and Ilhéus (12050’S-14050’S, 88-835 m. We recommend the species be uplisted to Endangered in the International Union for the Conservation of Nature’s global Red List of Threatened Species as its estimated extent of occurrence is under 5000km2. The effective conservation of B. pirajai will rely on addressing two key issues: improving our knowledge of the species and successfully mitigating habitat loss and fragmentation.

  15. New methodology for the obtainment of antibothropic factors from the South American opossum (Didelphis marsupialis) and jararaca snake (Bothrops jararaca).

    Science.gov (United States)

    Neves-Ferreira, A G; Valente, R H; Sá, P G; Rocha, S L; Moussatché, H; Domont, G B; Perales, J

    1999-10-01

    The antibothropic factor (ABF) from D. marsupialis was collected from perforated hollow plastic golf balls which were surgically implanted subcutaneously in anesthetized opossums, a technique originally described for the production of polyclonal antibodies. Two months after the implantation of the balls, approximately 15 ml of seromatous fluid from D. marsupialis (SFDm-50 mg total protein/ml) could be recovered monthly. Opossum serum as well as SFDm showed similar SDS-PAGE profiles and antihemorrhagic potencies against Bothrops jararaca snake venom (Bjv). The presence of ABF in SFDm was confirmed by immunoblotting, using rabbit polyclonal antibodies raised against ABF isolated from opossum serum. ABF isolated from SFDm or from serum by ion-exchange chromatography showed identical chromatographic and electrophoretic profiles. ABF fromboth sources displayed very similar antihemorrhagic and anticaseinolytic activities against Bjv. In the case of B. jararaca, polyethylene perforated tubes were inserted in the abdominal cavity and two months after implantation, approximately 4 ml of seromatous fluid from B. jararaca (SFBj-23 mg total protein/ml) were recovered. B.jararaca serum and SFBj showed the same native and SDS-PAGE band pattern. Both serum and SFBj inhibited Bjv hemorrhagic activity. We conclude that this new methodology is very suitable for continuously obtaining opossum ABF and SFBj, in large scale and in an easier way, avoiding animal suffering and eventual sacrifice. PMID:10414866

  16. Bothrops asper envenoming in cattle: Clinical features and management using equine-derived whole IgG antivenom.

    Science.gov (United States)

    Rodríguez, C; Estrada, R; Herrera, M; Gómez, A; Segura, Á; Vargas, M; Villalta, M; León, G

    2016-01-01

    Snakebite envenoming is an important problem in the livestock industry in Costa Rica. Of the 22 species of venomous snakes in the country, Bothrops asper is involved in most animal envenomings. Envenomation is typically characterised by swelling and bleeding at the bite site, coagulopathy, systemic haemorrhage, and, in some cases, death. The aims of the present study were to describe the clinical manifestations of B. asper envenomation in cattle and to evaluate the treatment efficacy of antivenom administration. The clinical effects of naturally occurring envenomation were reproduced experimentally in cattle by giving an intramuscular injection of either 10 mg or 50 mg venom to replicate mild and severe envenomings, respectively. Intravenous antivenom given 6 h after experimental venom injection controlled the symptoms; a dose of 120 mL was found to be appropriate for moderate and 200 mL for severe naturally occurring envenomings. Although administration of antivenom within the first 6 h following a snakebite prevented systemic effects, it did not reduce the extent of swelling at the bite site. Delayed administration of antivenom was not effective in saving naturally envenomed animals. The results indicate that, when promptly administered, antivenom constitutes an effective treatment for B. asper snakebite envenomation in cattle. PMID:27152384

  17. Isolation and characterization of two new non-hemorrhagic metalloproteinases with fibrinogenolytic activity from the mapanare (Bothrops colombiensis) venom.

    Science.gov (United States)

    Girón, María E; Rodríguez-Acosta, Alexis; Salazar, Ana María; Sánchez, Elda E; Galán, Jacob; Ibarra, Carlos; Guerrero, Belsy

    2013-01-01

    Colombienases are acidic, low molecular weight metalloproteinases (Mr of 23,074.31 Da colombienase-1 and 23,078.80 Da colombienase-2; pI of 6.0 and 6.2, respectively) isolated from Bothrops colombiensis snake venom. The chromatographic profile in RP-HPLC and its partial sequence confirmed its high homogeneity. Both colombienases present fibrino(geno)lytic activity, but did not show any hemorrhagic, amidolytic, plasminogen activator or coagulant activities, and no effect on platelet aggregation induced by collagen or ADP. Both enzymes were strongly active on fibrinogen Aα chains followed by the Bβ chains, and colombienases-2, at high doses, also degraded the γ chains. This activity was stable at temperatures ranging between 4 and 37 °C, with a maximum activity at 25 °C, and at pHs between 7 and 9. The homology demonstrated by the comparison of sequences, with zinc-dependent metalloproteinases, as well as the metal chelant effects on, confirmed that the colombienases were metalloproteinases, particularly to α-fibrinogenases belonging to the P-I class of SVPMs (20-30 kDa), which contain only the single-domain proteins. The biological characteristics of the colombienases confer a therapeutic potential, since they contain a high fibrino(geno)lytic activity, devoid of hemorrhagic activity. These metalloproteinases might be explored as thrombolytic agents given that they dissolve fibrin clots or prevent their formation.

  18. Identification of protective B-cell epitopes of Atroxlysin-I: A metalloproteinase from Bothrops atrox snake venom.

    Science.gov (United States)

    Schneider, F S; de Almeida Lima, S; Reis de Ávila, G; Castro, K L; Guerra-Duarte, C; Sanchez, E F; Nguyen, C; Granier, C; Molina, F; Chávez-Olortegui, C

    2016-03-29

    Atroxlysin-I (Atr-I) is a hemorrhagic snake venom metalloproteinase (SVMP) from Bothrops atrox venom, the snake responsible for the majority of bites in the north region of South America. SVMPs like Atr-I produce toxic effects in victims including hemorrhage, inflammation, necrosis and blood coagulation deficiency. Mapping of B-cell epitopes in SVMPs might result in the identification of non-toxic molecules capable of inducing neutralizing antibodies and improving the anti-venom therapy. Here, using the SPOT-synthesis technique we identified two epitopes located in the N-ter region of Atr-I (AtrEp1-(22)YNGNSDKIRRRIHQM(36); and AtrEp2-(55)GVEIWSNKDLINVQ(68)). Based on the sequence of AtrEp1 and AtrEp2 a third peptide named Atr-I biepitope (AtrBiEp) was designed and synthesized ((23)NGNSDKIRRRIH(34)GG(55)GVEIWSNKDLINVQ(68)). AtrBiEp was used to immunize BALB/c mice. Anti-AtrBiEp serum cross-reacted against Atr-I in western blot and was able to fully neutralize the hemorrhagic activity of Atr-I. Our results provide a rational basis for the identification of neutralizing epitopes on Atr-I snake venom toxin and show that the use of synthetic peptides could improve the generation of immuno-therapeutics. PMID:26917009

  19. Inflammatory mediators involved in the paw edema and hyperalgesia induced by Batroxase, a metalloproteinase isolated from Bothrops atrox snake venom.

    Science.gov (United States)

    De Toni, Lanuze G B; Menaldo, Danilo L; Cintra, Adélia C O; Figueiredo, Maria J; de Souza, Anderson R; Maximiano, William M A; Jamur, Maria C; Souza, Glória E P; Sampaio, Suely V

    2015-09-01

    Snake venom metalloproteinases have been described as responsible for several inflammatory effects. In this study, we investigated the edema and hyperalgesia induced in rats by Batroxase, a P-I metalloproteinase from Bothrops atrox venom, along with possible inflammatory mediators involved in these responses. Batroxase or sterile saline was injected into rat paws and the edema and hyperalgesic effects were evaluated for 6h by using a plethysmometer and a Von Frey system, respectively. Batroxase induced significant edematogenic and hyperalgesic peak responses in the first hours after administration. The inflammatory mediators involved in these responses were assayed by pretreatment of animals with synthesis inhibitors or receptor antagonists. Peak responses were significantly reduced by administration of the glucocorticoid dexamethasone, the H1 receptor antagonist diphenhydramine and the FLAP inhibitor MK-886. Rat paws injected with compound 48/80, a mast cell degranulating agent, followed by Batroxase injection resulted in significant reduction of the edema and hyperalgesia. However, Batroxase itself induced minor degranulation of RBL-2H3 mast cells in vitro. Additionally, the inflammatory responses did not seem to be related to prostaglandins, bradykinin or nitric oxide. Our results indicate a major involvement of histamine and leukotrienes in the edema and hyperalgesia induced by Batroxase, which could be related, at least in part, to mast cell degranulation. PMID:26072684

  20. Platelet-active substances in the venom of Bothrops moojeni snake-a novel evaluation method using whole blood aggregometry.

    Science.gov (United States)

    Demler, Christine; Bühler, Beatrice; Menin, Laure; Stöcklin, Reto; Wilmer, Marianne; Ernst, Beat; Perchuc, Anna Maria

    2010-01-01

    The objective of the present study was an investigation of the crude Bothrops moojeni venom, aiming at the identification of new compounds with platelet-activating or -inhibiting activity. The venom was separated by gel filtration chromatography into 18 fractions, which were tested by means of whole blood aggregometry for their activities affecting the aggregation of blood platelets. In order to eliminate interferences caused by prothrombin activators or thrombin like-enzymes, which are frequently present in snake venoms, a test method for screening protein mixtures was developed. To avoid clotting of the blood samples, the thrombin inhibitor hirudin and the synthetic inhibitor of fibrin polymerization Pefabloc FG were applied. In the present study, a platelet aggregation activator with an activity resembling thrombocytin from B. atrox was identified in one of the examined venom fractions. In addition, a platelet antagonist-most likely a disintegrin-with broad inhibitory activity against aggregation triggered by collagen, adenosine diphosphate and thrombin receptor activating peptide, was identified. PMID:19938887

  1. On the quaternary structure of a C-type lectin from Bothrops jararacussu venom--BJ-32 (BjcuL).

    Science.gov (United States)

    Silva, F P; Alexandre, G M C; Ramos, C H I; De-Simone, S G

    2008-12-15

    BJ-32 (also known as BjcuL) is a C-type lectin from the venom of Bothrops jararacussu with specificity for beta-galactosides and a remarkable ability to agglutinate several species of trypanosomatids. Our objective was to study the oligomerization state of native BJ-32 by using different biophysical and computational methods. Small-angle X-ray light scattering (SAXS) experiments disclosed a compact, globular protein with a radius of gyration of 36.72+/-0.04A and molecular weight calculated as 147.5+/-2.0kDa. From analytical ultracentrifugation analysis, it was determined that the BJ-32 sedimentation profile fits nicely to a decamer model. The analysis of the intrinsic emitted fluorescence spectra for BJ-32 solutions indicated that association of subunits in the decamer is accompanied by changes in the environment of Tryptophan residues. Both ab initio and comparative models of BJ-32 supported the resemblance of the decamer in the crystallographic structure from a close homologue, the rattlesnake venom lectin (RSL) from Crotalus atrox. PMID:18948130

  2. Purification from Bothrops lanceolatus (fer de lance) venom of a fibrino(geno)lytic enzyme with esterolytic activity.

    Science.gov (United States)

    Lôbo de Araújo, A; Donato, J L; Bon, C

    1998-05-01

    Bothrops lanceolatus venom has high caseinolytic, phospholipasic, esterolytic and hemorrhagic activities. In spite of having no coagulant effect on plasma, this venom contains a thrombin-like enzyme. Using gel filtration and ion-exchange chromatographies, we have purified an esterolytic fraction (F-II-1a) from this venom with a protein yield of 4% and a 58% recovery in enzyme activity. SDS-PAGE in the presence of beta-mercaptoethanol showed that the enzyme is a single chain polypeptide with a MW=38,100. Immunodiffusion and immunoelectrophoresis of fraction F-II-1a against serum from horses immunized with B. lanceolatus venom and against rabbit antiserum prepared using fraction F-II-1a both showed a single immunoprecipitin line. The Km and Vmax values for TAME hydrolysis were 0.85 mM and 38.6 micromol/min/mg, respectively. The esterolytic activity was completely inhibited by PMSF (10 mM) but not by EDTA (20 mM). Fraction F-II-1a hydrolyzed the alpha and beta chains of fibrinogen. Degradation of the alpha chain occurred within 10 min while that of the beta-chain was slower. The enzyme had no effect on the gamma-chain even after 4 h of hydrolysis. PMID:9655635

  3. Inhibition of the Myotoxicity Induced by Bothrops jararacussu Venom and Isolated Phospholipases A2 by Specific Camelid Single-Domain Antibody Fragments.

    Directory of Open Access Journals (Sweden)

    Nidiane D R Prado

    Full Text Available Antivenoms, produced using animal hyperimmune plasma, remains the standard therapy for snakebites. Although effective against systemic damages, conventional antivenoms have limited efficacy against local tissue damage. Additionally, the hypersensitivity reactions, often elicited by antivenoms, the high costs for animal maintenance, the difficulty of producing homogeneous lots, and the instability of biological products instigate the search for innovative products for antivenom therapy. In this study, camelid antibody fragments (VHH with specificity to Bothropstoxin I and II (BthTX-I and BthTX-II, two myotoxic phospholipases from Bothrops jararacussu venom, were selected from an immune VHH phage display library. After biopanning, 28 and 6 clones recognized BthTX-I and BthTX-II by ELISA, respectively. Complementarity determining regions (CDRs and immunoglobulin frameworks (FRs of 13 VHH-deduced amino acid sequences were identified, as well as the camelid hallmark amino acid substitutions in FR2. Three VHH clones (KF498607, KF498608, and KC329718 were capable of recognizing BthTX-I by Western blot and showed affinity constants in the nanomolar range against both toxins. VHHs inhibited the BthTX-II phospholipase A2 activity, and when tested for cross-reactivity, presented specificity to the Bothrops genus in ELISA. Furthermore, two clones (KC329718 and KF498607 neutralized the myotoxic effects induced by B. jararacussu venom, BthTX-I, BthTX-II, and by a myotoxin from Bothrops brazili venom (MTX-I in mice. Molecular docking revealed that VHH CDRs are expected to bind the C-terminal of both toxins, essential for myotoxic activity, and to epitopes in the BthTX-II enzymatic cleft. Identified VHHs could be a biotechnological tool to improve the treatment for snake envenomation, an important and neglected world public health problem.

  4. Inhibition of the Myotoxicity Induced by Bothrops jararacussu Venom and Isolated Phospholipases A2 by Specific Camelid Single-Domain Antibody Fragments

    Science.gov (United States)

    Prado, Nidiane D. R.; Pereira, Soraya S.; da Silva, Michele P.; Morais, Michelle S. S.; Kayano, Anderson M.; Moreira-Dill, Leandro S.; Luiz, Marcos B.; Zanchi, Fernando B.; Fuly, André L.; E. F. Huacca, Maribel; Fernandes, Cleberson F.; Calderon, Leonardo A.; Zuliani, Juliana P.; Soares, Andreimar M.; Stabeli, Rodrigo G.; F. C. Fernandes, Carla

    2016-01-01

    Antivenoms, produced using animal hyperimmune plasma, remains the standard therapy for snakebites. Although effective against systemic damages, conventional antivenoms have limited efficacy against local tissue damage. Additionally, the hypersensitivity reactions, often elicited by antivenoms, the high costs for animal maintenance, the difficulty of producing homogeneous lots, and the instability of biological products instigate the search for innovative products for antivenom therapy. In this study, camelid antibody fragments (VHH) with specificity to Bothropstoxin I and II (BthTX-I and BthTX-II), two myotoxic phospholipases from Bothrops jararacussu venom, were selected from an immune VHH phage display library. After biopanning, 28 and 6 clones recognized BthTX-I and BthTX-II by ELISA, respectively. Complementarity determining regions (CDRs) and immunoglobulin frameworks (FRs) of 13 VHH-deduced amino acid sequences were identified, as well as the camelid hallmark amino acid substitutions in FR2. Three VHH clones (KF498607, KF498608, and KC329718) were capable of recognizing BthTX-I by Western blot and showed affinity constants in the nanomolar range against both toxins. VHHs inhibited the BthTX-II phospholipase A2 activity, and when tested for cross-reactivity, presented specificity to the Bothrops genus in ELISA. Furthermore, two clones (KC329718 and KF498607) neutralized the myotoxic effects induced by B. jararacussu venom, BthTX-I, BthTX-II, and by a myotoxin from Bothrops brazili venom (MTX-I) in mice. Molecular docking revealed that VHH CDRs are expected to bind the C-terminal of both toxins, essential for myotoxic activity, and to epitopes in the BthTX-II enzymatic cleft. Identified VHHs could be a biotechnological tool to improve the treatment for snake envenomation, an important and neglected world public health problem. PMID:27028872

  5. Inhibition of the Myotoxicity Induced by Bothrops jararacussu Venom and Isolated Phospholipases A2 by Specific Camelid Single-Domain Antibody Fragments.

    Science.gov (United States)

    Prado, Nidiane D R; Pereira, Soraya S; da Silva, Michele P; Morais, Michelle S S; Kayano, Anderson M; Moreira-Dill, Leandro S; Luiz, Marcos B; Zanchi, Fernando B; Fuly, André L; Huacca, Maribel E F; Fernandes, Cleberson F; Calderon, Leonardo A; Zuliani, Juliana P; Pereira da Silva, Luiz H; Soares, Andreimar M; Stabeli, Rodrigo G; Fernandes, Carla F C

    2016-01-01

    Antivenoms, produced using animal hyperimmune plasma, remains the standard therapy for snakebites. Although effective against systemic damages, conventional antivenoms have limited efficacy against local tissue damage. Additionally, the hypersensitivity reactions, often elicited by antivenoms, the high costs for animal maintenance, the difficulty of producing homogeneous lots, and the instability of biological products instigate the search for innovative products for antivenom therapy. In this study, camelid antibody fragments (VHH) with specificity to Bothropstoxin I and II (BthTX-I and BthTX-II), two myotoxic phospholipases from Bothrops jararacussu venom, were selected from an immune VHH phage display library. After biopanning, 28 and 6 clones recognized BthTX-I and BthTX-II by ELISA, respectively. Complementarity determining regions (CDRs) and immunoglobulin frameworks (FRs) of 13 VHH-deduced amino acid sequences were identified, as well as the camelid hallmark amino acid substitutions in FR2. Three VHH clones (KF498607, KF498608, and KC329718) were capable of recognizing BthTX-I by Western blot and showed affinity constants in the nanomolar range against both toxins. VHHs inhibited the BthTX-II phospholipase A2 activity, and when tested for cross-reactivity, presented specificity to the Bothrops genus in ELISA. Furthermore, two clones (KC329718 and KF498607) neutralized the myotoxic effects induced by B. jararacussu venom, BthTX-I, BthTX-II, and by a myotoxin from Bothrops brazili venom (MTX-I) in mice. Molecular docking revealed that VHH CDRs are expected to bind the C-terminal of both toxins, essential for myotoxic activity, and to epitopes in the BthTX-II enzymatic cleft. Identified VHHs could be a biotechnological tool to improve the treatment for snake envenomation, an important and neglected world public health problem. PMID:27028872

  6. Collaborative Calibration of Control Standards of Hemocoagulase Bothrops Atrox%矛头蝮蛇血凝酶工作标准品的协作标定

    Institute of Scientific and Technical Information of China (English)

    李秀琳; 孙东; 丁忠福; 宁静; 于翀; 崔亮亮; 薛雁

    2016-01-01

    目的:标定我公司生产矛头蝮蛇血凝酶工作标准品。方法:以蛇毒血凝酶质量标准WS-349( X-317)-2001中效价测定为基础,改进测定方法,组织多名实验人员协作标定,并对数据进行统计分析。结果:本批矛头蝮蛇血凝酶工作标准品酶活性单位为103 U/支,加权均值标准差为0.8068,95%加权均值可信限的范围为101.0~105.1 U/支。结论:我公司生产的矛头蝮蛇血凝酶工作标准品标化科学、赋值准确,可以用于本公司产品的效价检测。%Objective:To calibrate the control standards of Hemocoagulase Bothrops atrox of our company.Methods:The enzyme ac-tivity was assayed based on the titration of Hemocoagulase Atrox Quality Standard WS-349(X-317)-2001, the method was modified, more than a number of experimenters were organized to collaboratively calibrate and the data was statistically analyzed.Results: The activity of the control standards of Hemocoagulase Bothrops atrox was 103 units per bottle, the weighted average standard deviation was 0.806 8, and the 95%weighted mean confidence range was 101.0-105.1 units per bottle.Conclusion:The calibration of the control standards of Hemocoagulase Bothrops atrox is scientific and accurate, and can be used to detect the titers of our company′s products .

  7. Clinical trial of two antivenoms for the treatment of Bothrops and Lachesis bites in the north eastern Amazon region of Brazil.

    Science.gov (United States)

    Pardal, Pedro Pereira de Oliveira; Souza, Suzana Medeiro; Monteiro, Maria Rita de Cássia da Costa; Fan, Hui Wen; Cardoso, João Luiz Costa; França, Francisco Oscar Siqueira; Tomy, Sandra Corrallo; Sano-Martins, Ida S; de Sousa-e-Silva, Maria Cristina Cirillo; Colombini, Mônica; Kodera, Nancy F; Moura-da-Silva, Ana Maria; Cardoso, Diva Ferreira; Velarde, David Toledo; Kamiguti, Aura S; Theakston, Robert David; Warrell, David A

    2004-01-01

    The efficacies of specific Bothrops atrox-Lachesis and standard Bothrops-Lachesis antivenoms were compared in the north eastern Amazon region of Brazil. The main aim was to investigate whether a specific antivenom raised against the venom of B. atrox, the most important Amazon snake species from a medical point of view, was necessary for the treatment of patients in this region. Seventy-four patients with local and systemic effects of envenoming by Bothrops or Lachesis snakes were randomly allocated to receive either specific (n = 38) or standard (n = 36) antivenoms. In 46 cases (24 in the standard antivenom group, 22 in the other) the snake was identified either by enzyme immunoassay or by examination of the dead snake, as B. atrox in 45, L. muta in one. Patients were similar in all clinical and epidemiological respects before treatment. Results indicated that both antivenoms were equally effective in reversing all signs of envenoming detected both clinically and in the laboratory. Venom-induced haemostatic abnormalities were resolved within 24 h after the start of antivenom therapy in most patients. The extent of local complications, such as local skin necrosis and secondary infection, was similar in both groups. There were no deaths. The incidence of early anaphylactic reactions was 18% and 19%, respectively for specific and standard antivenoms; none was life-threatening. Measurement of serum venom concentrations by enzyme immunoassay (EIA) confirmed that both antivenoms cleared venom antigenaemia effectively. EIA also revealed that one patient had been bitten by Lachesis muta, although the clinical features in this case were not distinctive. PMID:14702836

  8. Combined venomics, venom gland transcriptomics, bioactivities, and antivenomics of two Bothrops jararaca populations from geographic isolated regions within the Brazilian Atlantic rainforest.

    Science.gov (United States)

    Gonçalves-Machado, Larissa; Pla, Davinia; Sanz, Libia; Jorge, Roberta Jeane B; Leitão-De-Araújo, Moema; Alves, Maria Lúcia M; Alvares, Diego Janisch; De Miranda, Joari; Nowatzki, Jenifer; de Morais-Zani, Karen; Fernandes, Wilson; Tanaka-Azevedo, Anita Mitico; Fernández, Julián; Zingali, Russolina B; Gutiérrez, José María; Corrêa-Netto, Carlos; Calvete, Juan J

    2016-03-01

    Bothrops jararaca is a slender and semi-arboreal medically relevant pit viper species endemic to tropical and subtropical forests in southern Brazil, Paraguay, and northern Argentina (Misiones). Within its geographic range, it is often abundant and is an important cause of snakebite. Although no subspecies are currently recognized, geographic analyses have revealed the existence of two well-supported B. jararaca clades that diverged during the Pliocene ~3.8Mya and currently display a southeastern (SE) and a southern (S) Atlantic rainforest (Mata Atlântica) distribution. The spectrum, geographic variability, and ontogenetic changes of the venom proteomes of snakes from these two B. jararaca phylogroups were investigated applying a combined venom gland transcriptomic and venomic analysis. Comparisons of the venom proteomes and transcriptomes of B. jararaca from the SE and S geographic regions revealed notable interpopulational variability that may be due to the different levels of population-specific transcriptional regulation, including, in the case of the southern population, a marked ontogenetic venom compositional change involving the upregulation of the myotoxic PLA2 homolog, bothropstoxin-I. This population-specific marker can be used to estimate the proportion of venom from the southern population present in the B. jararaca venom pool used for the Brazilian soro antibotrópico (SAB) antivenom production. On the other hand, the southeastern population-specific D49-PLA2 molecules, BinTX-I and BinTX-II, lend support to the notion that the mainland ancestor of Bothrops insularis was originated within the same population that gave rise to the current SE B. jararaca phylogroup, and that this insular species endemic to Queimada Grande Island (Brazil) expresses a pedomorphic venom phenotype. Mirroring their compositional divergence, the two geographic B. jararaca venom pools showed distinct bioactivity profiles. However, the SAB antivenom manufactured in Vital Brazil

  9. Combined venomics, venom gland transcriptomics, bioactivities, and antivenomics of two Bothrops jararaca populations from geographic isolated regions within the Brazilian Atlantic rainforest.

    Science.gov (United States)

    Gonçalves-Machado, Larissa; Pla, Davinia; Sanz, Libia; Jorge, Roberta Jeane B; Leitão-De-Araújo, Moema; Alves, Maria Lúcia M; Alvares, Diego Janisch; De Miranda, Joari; Nowatzki, Jenifer; de Morais-Zani, Karen; Fernandes, Wilson; Tanaka-Azevedo, Anita Mitico; Fernández, Julián; Zingali, Russolina B; Gutiérrez, José María; Corrêa-Netto, Carlos; Calvete, Juan J

    2016-03-01

    Bothrops jararaca is a slender and semi-arboreal medically relevant pit viper species endemic to tropical and subtropical forests in southern Brazil, Paraguay, and northern Argentina (Misiones). Within its geographic range, it is often abundant and is an important cause of snakebite. Although no subspecies are currently recognized, geographic analyses have revealed the existence of two well-supported B. jararaca clades that diverged during the Pliocene ~3.8Mya and currently display a southeastern (SE) and a southern (S) Atlantic rainforest (Mata Atlântica) distribution. The spectrum, geographic variability, and ontogenetic changes of the venom proteomes of snakes from these two B. jararaca phylogroups were investigated applying a combined venom gland transcriptomic and venomic analysis. Comparisons of the venom proteomes and transcriptomes of B. jararaca from the SE and S geographic regions revealed notable interpopulational variability that may be due to the different levels of population-specific transcriptional regulation, including, in the case of the southern population, a marked ontogenetic venom compositional change involving the upregulation of the myotoxic PLA2 homolog, bothropstoxin-I. This population-specific marker can be used to estimate the proportion of venom from the southern population present in the B. jararaca venom pool used for the Brazilian soro antibotrópico (SAB) antivenom production. On the other hand, the southeastern population-specific D49-PLA2 molecules, BinTX-I and BinTX-II, lend support to the notion that the mainland ancestor of Bothrops insularis was originated within the same population that gave rise to the current SE B. jararaca phylogroup, and that this insular species endemic to Queimada Grande Island (Brazil) expresses a pedomorphic venom phenotype. Mirroring their compositional divergence, the two geographic B. jararaca venom pools showed distinct bioactivity profiles. However, the SAB antivenom manufactured in Vital Brazil

  10. Pharmacological characterisation of arthritis induced by Bothrops jararaca venom in rabbits: a positive cross talk between bradykinin, nitric oxide and prostaglandin E2.

    OpenAIRE

    Suzana B. V. Mello; Maria Luiza Guzzo; Luiz Filipe Santiago Lisboa; Farsky, Sandra H P

    2002-01-01

    BACKGROUND: Our previous results showed that nitric oxide (NO) and bradykinin (BK) mediate the arthritis induced by Bothrops jararaca venom (BjV) in rabbits. In this study, we investigated the contribution of each receptor of BK as well as the inter-relationship between NO and eicosanoids in BjV-induced arthritis. METHODS: The arthritis was induced in rabbits with 16 microg of BjV injected intra-articularly. Prostaglandin E2 (PGE2), thromboxane B2 (TxB2), leukotriene B4 (LTB4) (radioimmunoass...

  11. ESI-MS/MS Identification of a Bradykinin-Potentiating Peptide from Amazon Bothrops atrox Snake Venom Using a Hybrid Qq-oaTOF Mass Spectrometer

    OpenAIRE

    Calderon, Leonardo A.; Rodrigo G. Stábeli; Juliana P Zuliani; Soares, Andreimar M.; Anderson M. Kayano; Silva, Rodrigo S.; Zaqueo, Kayena D.; Souza, Gustavo H. M. F.; Cleópatra A. S. Caldeira; Antonio Coutinho-Neto

    2013-01-01

    A bradykinin-potentiating peptide (BPP) from Amazon Bothrops atrox venom with m/z 1384.7386 was identified and characterized by collision induced dissociation (CID) using an ESI-MS/MS spectra obtained in positive ion mode on a hybrid Qq-oaTOF mass spectrometer, Xevo G2 QTof MS (Waters, Manchester, UK). D e novo peptide sequence analysis of the CID fragmentation spectra showed the amino acid sequence ZKWPRPGPEIPP, with a pyroglutamic acid and theoretical monoisotopic m/z 1384.7378, which is si...

  12. An alternative method to isolate protease and phospholipase A2 toxins from snake venoms based on partitioning of aqueous two-phase systems

    Directory of Open Access Journals (Sweden)

    GN Gómez

    2012-01-01

    Full Text Available Snake venoms are rich sources of active proteins that have been employed in the diagnosis and treatment of health disorders and antivenom therapy. Developing countries demand fast economical downstream processes for the purification of this biomolecule type without requiring sophisticated equipment. We developed an alternative, simple and easy to scale-up method, able to purify simultaneously protease and phospholipase A2 toxins from Bothrops alternatus venom. It comprises a multiple-step partition procedure with polyethylene-glycol/phosphate aqueous two-phase systems followed by a gel filtration chromatographic step. Two single bands in SDS-polyacrylamide gel electrophoresis and increased proteolytic and phospholipase A2 specific activities evidence the homogeneity of the isolated proteins.

  13. Neutralization of pharmacological and toxic activities of Bothrops jararacussu snake venom and isolated myotoxins by Serjania erecta methanolic extract and its fractions

    Directory of Open Access Journals (Sweden)

    RS Fernandes

    2011-01-01

    Full Text Available Most of the snakebites recorded in Brazil are caused by the Bothrops genus. Given that the local tissue damage caused by this genus cannot be treated by antivenom therapy, numerous studies are focusing on supplementary alternatives, such as the use of medicinal plants. Serjania erecta has already demonstrated anti-inflammatory, antiseptic and healing properties. In the current study, the aerial parts of S. erecta were extracted with methanol, then submitted to chromatographic fractionation on a Sephadex LH20 column and eluted with methanol, which resulted in four main fractions. The crude extract and fractions neutralized the toxic activities of Bothrops jararacussu snake venom and isolated myotoxins (BthTX-I and II. Results showed that phospholipase A2, fibrinogenolytic, myotoxic and hemorrhagic activities were inhibited by the extract. Moreover, the myotoxic and edematous activities induced by BthTX-I, and phospholipase A2 activity induced by BthTX-II, were inhibited by the extract of S. erecta and its fraction. The clotting time on bovine plasma was significantly prolonged by the inhibitory action of fractions SF3 and SF4. This extract is a promising source of natural inhibitors, such as flavonoids and tannins, which act by forming complexes with metal ions and proteins, inhibiting the action of serineproteases, metalloproteases and phospholipases A2.

  14. Structural and biophysical studies with the MjTX-I, a Lys49-phospholipase A{sub 2} homologue from Bothrops moojeni venom

    Energy Technology Data Exchange (ETDEWEB)

    Salvador, G.H.M.; Fernandes, C.A.H.; Fernandez, R.M.; Fontes, M.R.M. [UNESP, Universidade Estadual Paulista, Botucatu, SP (Brazil); Marchi-Salvador, D.P. [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Soares, A.M. [Universidade de Sao Paulo (USP-RP), Ribeirao Preto, SP (Brazil); Oliveira, C.L.P [Universidade de Sao Paulo (USP), SP (Brazil)

    2012-07-01

    Full text: Phospholipases A{sub 2} (PLA{sub 2}) are small proteins found in a great diversity of organisms and belong to a superfamily of proteins involved in many important pharmacological processes, such as neurotoxicity, myotoxicity, platelet aggregation, and anticoagulant activity. Ophidic accidents caused by snakes from Bothrops genus are not efficiently neutralized by conventional serum therapy, and then detailed studies with this class of proteins may be very important to supplement this conventional therapy. Miotoxin-I (MjTX-I) is a basic Lys49-PLA{sub 2}, isolated from Bothrops moojeni snake venom, which induces a drastic local myonecrosis. Crystal structure of MjTX-I shows four molecules in the asymmetric unit, an unusually oligomeric conformation for snake venom Lys49-PLA{sub 2}s. However, bioinformatics techniques indicate a dimer as the biological oligomeric conformation. To get additional information of its biological conformation, we also performed Dynamic Light Scattering, Size Exclusion Chromatography and Small Angle X-ray Scattering experiments. These techniques showed a monomer as the most probable biological conformation in water; however small changes in pH and ionic strength result in different oligomeric assemblies. These novel information for Lys49-PLA{sub 2}s may result in important conclusions for this intriguing class of toxins. (author)

  15. Acidentes ofidicos causados por Bothrops moojeni: correlação do quadro clínico com o tamanho da serpente

    Directory of Open Access Journals (Sweden)

    João Aris Kouyoumdjian

    1989-04-01

    Full Text Available Durante um período de 19 meses (março 1986 a setembro 1987 foram estudados 22 casos de acidentes ofídicos causados por Bothrops moojeni na região de São José do Rio Preto, São Paulo, nos quais o tamanho da serpente foi sistematicamente medido. Foram constituídos dois grupos de pacientes de acordo com o tamanho da serpente: grupo I - 9 casos de serpentes pequenas (30 a 53 cm e grupo II - 13 casos de serpentes grandes (80 a 147 cm. Os resultados mostraram: 1. efeitos locais iniciais - dor e edema - mais leves no grupo I; 2. tempo de coagulação prolongado/incoagulável levemente mais freqüente no grupo I; 3. complicações locais - necrose, infecção e síndrome compartimentai - exclusivamente, e em mais da metade dos casos do grupo II, apesar da terapia com antiveneno ter sido mais rápida e em doses maiores nesse grupo. Conclui-se que as serpentes Bothrops moojeni maiores apresentam grande incremento nas suas ações locais - edema, necrose e infecção secundária - e leve perda em sua ação coagulante.

  16. Isotopic change in the tissues of Bothrops atrox in captivity collected from environments of the eastern Amazon

    Science.gov (United States)

    Martinez, M. G.; Chalkidis, H. D.; Amazonas, D. R.; da Silva, A. M.; De Oliveira, R., Jr.; Camargo, P. B.

    2013-12-01

    The Bothrops atrox is little studied because it is sympatric Amazonian animals, and very little is known about the ecology and natural history of this species. It has a generalist diet and the distribution of this species is very wide. The adult animals forage mostly on the ground, while the younger animals prefer to stay on the vegetation. They are easily find in the rainy months in areas near lakes and seasonally flooded and are difficult to find in the driest months, a period where there is less availability of preys in these environments. Due to its aggressiveness, is considered one of the most feared snakes in South America and in the eastern Amazon, being responsible for the largest number of snakebites in the region. Through stable isotope carbon-13 and nitrogen-15, is intended to characterize the variations of the feeding habits of these collected animals in different environments and also when they are kept in captivity, feeding the animal's bioterium. The serpents were collected in environments with different land uses, such as native forest, savannah, pasture and have been brought to the serpentarium Integrated College Tapajos (FIT), being retained in order to Samplings throughout the experiment with feeding mice's own bioterium. When these snakes came from different locations, samples were collected scales and blood (T0), before receiving the new supply (captive), and every time we fed the mice the vivarium, new tissue samples were collected, (T1, T2, T3) to exchange all the nature of food for the food captivity.Based on the results of δ13C and δ15N, the samples collected in the tissues of snakes of different environments (nature and captivity), it was observed that changes in food sources reflect changes in tissues (blood and scales), also reflecting the production of poison different periods of turnover of absorbed material in those tissues, contributing to the study of animal ecology and behavior in relation to habitat.

  17. Poor regenerative outcome after skeletal muscle necrosis induced by Bothrops asper venom: alterations in microvasculature and nerves.

    Directory of Open Access Journals (Sweden)

    Rosario Hernández

    Full Text Available BACKGROUND: Viperid snakebite envenoming is characterized by prominent local tissue damage, including muscle necrosis. A frequent outcome of such local pathology is deficient skeletal muscle regeneration, which causes muscle dysfunction, muscle loss and fibrosis, thus provoking permanent sequelae that greatly affect the quality of life of patients. The causes of such poor regenerative outcome of skeletal muscle after viperid snakebites are not fully understood. METHODOLOGY/PRINCIPAL FINDINGS: A murine model of muscle necrosis and regeneration was adapted to study the effects of the venom and isolated toxins of Bothrops asper, the medically most important snake in Central America. Gastrocnemius muscle was injected with either B. asper venom, a myotoxic phospholipase A(2 (Mtx, a hemorrhagic metalloproteinase (SVMP, or saline solution. At various time intervals, during one month, tissue samples were collected and analyzed by histology, and by immunocytochemical and immunohistochemical techniques aimed at detecting muscle fibers, collagen, endothelial cells, myoblasts, myotubes, macrophages, TUNEL-positive nuclei, and axons. A successful regenerative response was observed in muscle injected with Mtx, which induces myonecrosis but does not affect the microvasculature. In contrast, poor regeneration, with fibrosis and atrophic fibers, occurred when muscle was injected with venom or SVMP, both of which provoke necrosis, microvascular damage leading to hemorrhage, and poor axonal regeneration. CONCLUSIONS/SIGNIFICANCE: The deficient skeletal muscle regeneration after injection of B. asper venom is likely to depend on the widespread damage to the microvasculature, which affects the removal of necrotic debris by phagocytes, and the provision of nutrients and oxygen required for regeneration. In addition, deficient axonal regeneration is likely to contribute to the poor regenerative outcome in this model.

  18. Snakebites and ethnobotany in the northwest region of Colombia. Part III: neutralization of the haemorrhagic effect of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Barona, J; Fonnegra, R; Jiménez, S L; Osorio, R G; Saldarriaga, M; Díaz, A

    2000-11-01

    Thirty-one of 75 extracts of plants used by traditional healers for snakebites, had moderate or high neutralizing ability against the haemorrhagic effect of Bothrops atrox venom from Antioquia and Chocó, north-western Colombia. After preincubation of several doses of every extract (7.8-4000 microg/mouse) with six minimum haemorrhagic doses (10 microg) of venom, 12 of them demonstrated 100% neutralizing capacity when the mixture was i.d. injected into mice (18-20 g). These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plants of Pleopeltis percussa (Polypodiaceae), Trichomanes elegans (Hymenophyllaceae) and Senna dariensis (Caesalpiniaceae); rhizomes of Heliconia curtispatha (Heliconiaceae); leaves and branches of Bixa orellana (Bixaceae), Philodendron tripartitum (Araceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the ripe fruits of Citrus limon (Rutaceae); leaves, branches and stem of Ficus nymphaeifolia (Moraceae). Extracts of another 19 species showed moderate neutralization (21-72%) at doses up to 4 mg/mouse, e.g. the whole plants of Aristolochia grandiflora (Aristolochiaceae), Columnea kalbreyeriana (Gesneriaceae), Sida acuta (Malvaceae), Selaginella articulata (Selaginellaceae) and Pseudoelephantopus spicatus (Asteraceae); rhizomes of Renealmia alpinia (Zingiberaceae); the stem of Strychnos xinguensis (Loganiaceae); leaves, branches and stems of Hyptis capitata (Lamiaceae), Ipomoea cairica (Convolvulaceae), Neurolaena lobata (Asteraceae), Ocimum micranthum (Lamiaceae), Piper pulchrum (Piperaceae), Siparuna thecaphora (Monimiaceae), Castilla elastica (Moraceae) and Allamanda cathartica (Apocynaceae); the macerated ripe fruits of Capsicum frutescens (Solanaceae); the unripe fruits of Crescentia cujete (Bignoniaceae); leaves and branches of Piper arboreum (Piperaceae) and Passiflora quadrangularis (Passifloraceae). When the extracts were independently administered

  19. Inhibition of toxic activities of Bothrops asper venom and other crotalid snake venoms by a novel neutralizing mixture.

    Science.gov (United States)

    Borkow, G; Gutierrez, J M; Ovadia, M

    1997-12-01

    The majority of snake bites in Central America are caused by Bothrops asper, whose venom induce complex local effects such as myonecrosis, edema and especially hemorrhage. These effects are only partially neutralized by the clinically used antivenom, even when administered rapidly after envenomation. Recently we screened 49 substances for antihemorrhagic activity and found that a mixture composed of CaNa2, EDTA, a B. asper serum fraction (natural antidote), and the currently used horse polyvalent antiserum is highly effective in the neutralization of local and systemic hemorrhage developing after B. asper envenomation (Borkow et al., Toxicon 35, 865-877, 1997). In the present study we screened the best six antihemorrhagic compounds for their capacity to neutralize the lethal activity in mice and the proteolytic, hemolytic, and antiattachment activities in vitro of the venom. The compounds tested included the currently used horse antivenom, rabbit antiserum against whole B. asper venom or against heated venom, B. asper and Natrix tessellata serum fractions, and CaNa2 EDTA. The constituents of the antihemorrhagic mixture were also the best inhibitors of the other examined toxic activities. Importantly, the mixture effectively neutralized toxic activities of an additional nine venoms from snakes abundant in Central America. This work suggests that the polyvalent antivenom used in Central America could be enriched with a B. asper serum fraction producing a more effective antivenom. In addition, the local application of CaNa2 EDTA to neutralize hemorrhagic toxins, immediately after a snake bite, may provide rapid inhibition of local damage caused by the venoms. PMID:9439739

  20. Snakebites and ethnobotany in the northwest region of Colombia. Part III: neutralization of the haemorrhagic effect of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Barona, J; Fonnegra, R; Jiménez, S L; Osorio, R G; Saldarriaga, M; Díaz, A

    2000-11-01

    Thirty-one of 75 extracts of plants used by traditional healers for snakebites, had moderate or high neutralizing ability against the haemorrhagic effect of Bothrops atrox venom from Antioquia and Chocó, north-western Colombia. After preincubation of several doses of every extract (7.8-4000 microg/mouse) with six minimum haemorrhagic doses (10 microg) of venom, 12 of them demonstrated 100% neutralizing capacity when the mixture was i.d. injected into mice (18-20 g). These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plants of Pleopeltis percussa (Polypodiaceae), Trichomanes elegans (Hymenophyllaceae) and Senna dariensis (Caesalpiniaceae); rhizomes of Heliconia curtispatha (Heliconiaceae); leaves and branches of Bixa orellana (Bixaceae), Philodendron tripartitum (Araceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the ripe fruits of Citrus limon (Rutaceae); leaves, branches and stem of Ficus nymphaeifolia (Moraceae). Extracts of another 19 species showed moderate neutralization (21-72%) at doses up to 4 mg/mouse, e.g. the whole plants of Aristolochia grandiflora (Aristolochiaceae), Columnea kalbreyeriana (Gesneriaceae), Sida acuta (Malvaceae), Selaginella articulata (Selaginellaceae) and Pseudoelephantopus spicatus (Asteraceae); rhizomes of Renealmia alpinia (Zingiberaceae); the stem of Strychnos xinguensis (Loganiaceae); leaves, branches and stems of Hyptis capitata (Lamiaceae), Ipomoea cairica (Convolvulaceae), Neurolaena lobata (Asteraceae), Ocimum micranthum (Lamiaceae), Piper pulchrum (Piperaceae), Siparuna thecaphora (Monimiaceae), Castilla elastica (Moraceae) and Allamanda cathartica (Apocynaceae); the macerated ripe fruits of Capsicum frutescens (Solanaceae); the unripe fruits of Crescentia cujete (Bignoniaceae); leaves and branches of Piper arboreum (Piperaceae) and Passiflora quadrangularis (Passifloraceae). When the extracts were independently administered

  1. Hemorrhagic, coagulant and fibrino(geno)lytic activities of crude venom and fractions from mapanare (Bothrops colombiensis) snakes.

    Science.gov (United States)

    Girón, María E; Salazar, Ana M; Aguilar, Irma; Pérez, John C; Sánchez, Elda E; Arocha-Piñango, Carmen L; Rodríguez-Acosta, Alexis; Guerrero, Belsy

    2008-01-01

    Bothrops colombiensis venom from two similar geographical locations were tested for their hemostatic functions and characterized by gel-filtration chromatography and SDS-PAGE electrophoresis. The snakes were from Caucagua and El Guapo towns of the Venezuelan state of Miranda. Fibrino(geno)lytic, procoagulant, hemorrhagic, lethal activities, gel-filtration chromatography and SDS-PAGE profiles were analyzed and compared for both venoms. The highest hemorrhagic activity of 5.3 mug was seen in El Guapo venom while Caucagua venom had the lowest LD(50) of 5.8 mg/kg. Both venoms presented similar thrombin-like activity. El Guapo showed a factor Xa-like activity two times higher than Caucagua. Differences were observed in kallikrein-like and t-PA activities, being highest in El Guapo. Caucagua venom showed the maximum fibrin lysis. Both crude venom runs on Sephadex G-100 chromatography gave fraction SII with the high fibrinolytic activity. Proteases presented in SII fractions and eluted from Benzamidine-Sepharose (not bound to the column) provoked a fast degradation of fibrinogen alpha chains and a slower degradation of beta chains, which could possibly be due to a higher content of alpha fibrinogenases in these venoms. The fibrinogenolytic activity was decreased by metalloprotease inhibitors. The results suggested that metalloproteases in SII fractions were responsible for the fibrinolytic activity. The analysis of samples for fibrin-zymography of SII fractions showed an active band with a molecular mass of approximately 30 kDa. These results reiterate the importance of using pools of venoms for antivenom immunization, to facilitate the neutralization of the maximum potential number of toxins.

  2. Cell cycle arrest evidence, parasiticidal and bactericidal properties induced by L-amino acid oxidase from Bothrops atrox snake venom.

    Science.gov (United States)

    de Melo Alves Paiva, Raquel; de Freitas Figueiredo, Raquel; Antonucci, Gilmara Ausech; Paiva, Helder Henrique; de Lourdes Pires Bianchi, Maria; Rodrigues, Kelly C; Lucarini, Rodrigo; Caetano, Renato Cesar; Linhari Rodrigues Pietro, Rosemeire Cristina; Gomes Martins, Carlos Henrique; de Albuquerque, Sérgio; Sampaio, Suely Vilela

    2011-05-01

    The present article describes an l-amino acid oxidase from Bothrops atrox snake venom as with antiprotozoal activities in Trypanosoma cruzi and in different species of Leishmania (Leishmania braziliensis, Leishmania donovani and Leishmania major). Leishmanicidal effects were inhibited by catalase, suggesting that they are mediated by H(2)O(2) production. Leishmania spp. cause a spectrum of diseases, ranging from self-healing ulcers to disseminated and often fatal infections, depending on the species involved and the host's immune response. BatroxLAAO also displays bactericidal activity against both Gram-positive and Gram-negative bacteria. The apoptosis induced by BatroxLAAO on HL-60 cell lines and PBMC cells was determined by morphological cell evaluation using a mix of fluorescent dyes. As revealed by flow cytometry analysis, suppression of cell proliferation with BatroxLAAO was accompanied by the significant accumulation of cells in the G0/G1 phase boundary in HL-60 cells. BatroxLAAO at 25 μg/mL and 50 μg/mL blocked G0-G1 transition, resulting in G0/G1 phase cell cycle arrest, thereby delaying the progression of cells through S and G2/M phase in HL-60 cells. This was shown by an accentuated decrease in the proportion of cells in S phase, and the almost absence of G2/M phase cell population. BatroxLAAO is an interesting enzyme that provides a better understanding of the ophidian envenomation mechanism, and has biotechnological potential as a model for therapeutic agents. PMID:21300133

  3. Biotechnological application of protein Leuc-B isolated from Bothrops leucurus venom as a prototype for antitumoral radiopharmaceutical;Aplicacao biotecnologica da proteina Leuc-B isolada da peconha de Bothrops leucurus como prototipo de radiofarmaco antitumoral

    Energy Technology Data Exchange (ETDEWEB)

    Gabriel, Lucilene Marcia

    2010-07-01

    According to the report of the International Agency for Research on Cancer, the growth of this disease implies the death of 17 million people a year by 2030. Although the knowledge on development of cancer is growing considerably, just a few advances in the diagnosis and therapy has been achieved. Faced with this scenario, it is clear the need for new substances more specifics with low toxicity to the patient, which can be used for diagnosis and treatment of cancer. Membrane receptors over expressed in tumor cells are promising target candidates for development of diagnostic and therapeutical tools. Integrins are a family of hetero dimeric cell surface adhesion receptors able to recognize and bind to proteins in the extracellular matrix (ECM). This recognition is mainly through the RGD domains presents in both the cell surface as in the protein from the ECM. Various integrins have been identified as regulators of tumor progression. The RGD domain is also found in some snake venoms named disintegrins. Disintegrins inhibit cell-matrix and a cell-cell interactions mediated by integrins and it has been shown that these proteins are able to inhibit metastasis in processes dependent on integrin. The disintegrin-like (ECD), as well as RGD-disintegrin are also able to bind to cell surface integrins and inhibit their adhesion to the natural ligands. In this work it was purified from Bothrops leucurus venom (VBL), a metalloproteinase-class P-III with disintegrin-like domain (ECD), Leucurolisina B (Leuc-B). This metalloproteinase and the crude venom were used to evaluate their applicability in the differential detection of tumors. In vitro results demonstrated that both VBL and Leuc-B have potent antitumoral effect on several cancer cell lines: U87, T98, RT2 (glioblastoma), MCF7 (breast), Ehrlich and UACC (melanoma) with IC{sub 50} values of approximately 0.6 muM. The morphological changes observed in these strains when treated with Leuc-B, and data from the DAPI staining

  4. Biotechnological application of protein Leuc-B isolated from Bothrops leucurus venom as a prototype for antitumoral radiopharmaceutical

    International Nuclear Information System (INIS)

    According to the report of the International Agency for Research on Cancer, the growth of this disease implies the death of 17 million people a year by 2030. Although the knowledge on development of cancer is growing considerably, just a few advances in the diagnosis and therapy has been achieved. Faced with this scenario, it is clear the need for new substances more specifics with low toxicity to the patient, which can be used for diagnosis and treatment of cancer. Membrane receptors over expressed in tumor cells are promising target candidates for development of diagnostic and therapeutical tools. Integrins are a family of hetero dimeric cell surface adhesion receptors able to recognize and bind to proteins in the extracellular matrix (ECM). This recognition is mainly through the RGD domains presents in both the cell surface as in the protein from the ECM. Various integrins have been identified as regulators of tumor progression. The RGD domain is also found in some snake venoms named disintegrins. Disintegrins inhibit cell-matrix and a cell-cell interactions mediated by integrins and it has been shown that these proteins are able to inhibit metastasis in processes dependent on integrin. The disintegrin-like (ECD), as well as RGD-disintegrin are also able to bind to cell surface integrins and inhibit their adhesion to the natural ligands. In this work it was purified from Bothrops leucurus venom (VBL), a metalloproteinase-class P-III with disintegrin-like domain (ECD), Leucurolisina B (Leuc-B). This metalloproteinase and the crude venom were used to evaluate their applicability in the differential detection of tumors. In vitro results demonstrated that both VBL and Leuc-B have potent antitumoral effect on several cancer cell lines: U87, T98, RT2 (glioblastoma), MCF7 (breast), Ehrlich and UACC (melanoma) with IC50 values of approximately 0.6 μM. The morphological changes observed in these strains when treated with Leuc-B, and data from the DAPI staining solution

  5. Diversity of metalloproteinases in Bothrops neuwiedi snake venom transcripts: evidences for recombination between different classes of SVMPs

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    Valente Richard H

    2011-11-01

    Full Text Available Abstract Background Snake venom metalloproteinases (SVMPs are widely distributed in snake venoms and are versatile toxins, targeting many important elements involved in hemostasis, such as basement membrane proteins, clotting proteins, platelets, endothelial and inflammatory cells. The functional diversity of SVMPs is in part due to the structural organization of different combinations of catalytic, disintegrin, disintegrin-like and cysteine-rich domains, which categorizes SVMPs in 3 classes of precursor molecules (PI, PII and PIII further divided in 11 subclasses, 6 of them belonging to PII group. This heterogeneity is currently correlated to genetic accelerated evolution and post-translational modifications. Results Thirty-one SVMP cDNAs were full length cloned from a single specimen of Bothrops neuwiedi snake, sequenced and grouped in eleven distinct sequences and further analyzed by cladistic analysis. Class P-I and class P-III sequences presented the expected tree topology for fibrinolytic and hemorrhagic SVMPs, respectively. In opposition, three distinct segregations were observed for class P-II sequences. P-IIb showed the typical segregation of class P-II SVMPs. However, P-IIa grouped with class P-I cDNAs presenting a 100% identity in the 365 bp at their 5' ends, suggesting post-transcription events for interclass recombination. In addition, catalytic domain of P-IIx sequences segregated with non-hemorrhagic class P-III SVMPs while their disintegrin domain grouped with other class P-II disintegrin domains suggesting independent evolution of catalytic and disintegrin domains. Complementary regions within cDNA sequences were noted and may participate in recombination either at DNA or RNA levels. Proteins predicted by these cDNAs show the main features of the correspondent classes of SVMP, but P-IIb and P-IIx included two additional cysteines cysteines at the C-termini of the disintegrin domains in positions not yet described. Conclusions In

  6. Digenetic trematodes infection in a Bothrops moojeni (Viperidae population from a fauna rescue in Porto Primavera, São Paulo State

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    Barrella T.H.

    2003-01-01

    Full Text Available No presente estudo é descrita a infecção por trematódeos digenéticos parasitas da cavidade oral e esôfago em uma população de serpentes Bothrops moojeni provenientes de resgate de fauna em Porto Primavera, Estado de São Paulo. Foi observada prevalência de infecção de 68%. O grau de infecção (número de trematódeos por serpente variou de 2 a 51 helmintos. Os trematódeos encontrados foram Ophisthogonimus spp. e Sticholecitha serpentis. A alta prevalência de infecção foi associada com a drástica alteração ambiental e o estresse multi-fatorial aos quais os animais foram submetidos, que poderiam ter favorecido o ciclo dos parasitas.

  7. Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil

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    Roger Maia Dias Ledo

    2012-09-01

    Full Text Available Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil. This paper presents a list of necrophagous insects associated with small size carrions of two reptiles and one amphibian, found in areas of riparian forests and Cerrado sensu stricto physiognomies in a Conservation Unit located in Brasilia, Distrito Federal. We found seven species of insects related to these carcasses, being five Sarcophagidae, one Calliphoridae and one Braconidae parasitoid wasp. Lucilia eximia and Peckia (Pattonella intermutans were the most abundant species in the study, corroborating with other studies that suggests that these species have specializations for colonization of small size animal carcasses.

  8. An Isoflavone from Dipteryx alata Vogel is Active against the in Vitro Neuromuscular Paralysis of Bothrops jararacussu Snake Venom and Bothropstoxin I, and Prevents Venom-Induced Myonecrosis

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    Miriéle C. Ferraz

    2014-05-01

    Full Text Available Snakebite is a neglected disease and serious health problem in Brazil, with most bites being caused by snakes of the genus Bothrops. Although serum therapy is the primary treatment for systemic envenomation, it is generally ineffective in neutralizing the local effects of these venoms. In this work, we examined the ability of 7,8,3'-trihydroxy-4'-methoxyisoflavone (TM, an isoflavone from Dipteryx alata, to neutralize the neurotoxicity (in mouse phrenic nerve-diaphragm preparations and myotoxicity (assessed by light microscopy of Bothrops jararacussu snake venom in vitro. The toxicity of TM was assessed using the Salmonella microsome assay (Ames test. Incubation with TM alone (200 μg/mL did not alter the muscle twitch tension whereas incubation with venom (40 μg/mL caused irreversible paralysis. Preincubation of TM (200 μg/mL with venom attenuated the venom-induced neuromuscular blockade by 84% ± 5% (mean ± SEM; n = 4. The neuromuscular blockade caused by bothropstoxin-I (BthTX-I, the major myotoxic PLA2 of this venom, was also attenuated by TM. Histological analysis of diaphragm muscle incubated with TM showed that most fibers were preserved (only 9.2% ± 1.7% were damaged; n = 4 compared to venom alone (50.3% ± 5.4% of fibers damaged; n = 3, and preincubation of TM with venom significantly attenuated the venom-induced damage (only 17% ± 3.4% of fibers damaged; n = 3; p < 0.05 compared to venom alone. TM showed no mutagenicity in the Ames test using Salmonella strains TA98 and TA97a with (+S9 and without (−S9 metabolic activation. These findings indicate that TM is a potentially useful compound for antagonizing the neuromuscular effects (neurotoxicity and myotoxicity of B. jararacussu venom.

  9. Caracterización de la enzima similar a trombina del veneno de Bothrops pictus "jergón de costa

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    Dan Vivas-Ruiz

    2015-12-01

    Full Text Available Objetivos. Realizar una caracterización bioquímica y molecular del principio coagulante del veneno de Bothrops pictus. Materiales y métodos. Se realizó la amplificación del gen a partir de cDNA, se analizó la homología de la secuencia nucleotídica y de la proteína deducida. Se procedió a purificar la enzima para los análisis de secuenciación directa N terminal de los primeros 20 aminoácidos y los ensayos de coagulación sobre plasma humano y fibrinógeno humano, por otro lado, se evaluó el patrón de corte del fibrinógeno por medio de PAGE SDS y la actividad defibrinogenante en roedores albinos (18-22 g. Se determinó el contenido de carbohidratos asociados, el efecto de inhibidores clásicos de proteasas y el efecto de iones bajo la forma de cloruros. Resultados. La enzima mostró homología en la estructura primaria con otras TLEs reportadas para la familia Viperidae, la dosis coagulante mínima (DCM sobre plasma y fibrinógeno humano fue de 18 y 6 µg respectivamente y su potencia coagulante fue de 131,1 NHI unidades de trombina. La enzima se mostró estable a condiciones fisiológicas y prescinde de iones para su actividad. Los carbohidratos asociados detectados fueron hexosas (25,76%, hexosaminas (13,1% y ácido siálico (0,76%. Los agentes fluoruro de fenil metil sulfonil floruro (PMSF ditiotreitol (DTT fueron los principales inhibidores de la actividad enzimática en tanto que la heparina no tuvo efecto inhibidor. Conclusiones. El principio coagulante del veneno de Bothrops pictus es una enzima similar a trombina

  10. Caracterización de la enzima similar a trombina del veneno de Bothrops pictus "jergón de costa

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    Dan Vivas-Ruiz

    2015-10-01

    Full Text Available Objetivos. Realizar una caracterización bioquímica y molecular del principio coagulante del veneno de Bothrops pictus. Materiales y métodos. Se realizó la amplificación del gen a partir de cDNA, se analizó la homología de la secuencia nucleotídica y de la proteína deducida. Se procedió a purificar la enzima para los análisis de secuenciación directa N terminal de los primeros 20 aminoácidos y los ensayos de coagulación sobre plasma humano y fibrinógeno humano, por otro lado, se evaluó el patrón de corte del fibrinógeno por medio de PAGE SDS y la actividad defibrinogenante en roedores albinos (18-22 g. Se determinó el contenido de carbohidratos asociados, el efecto de inhibidores clásicos de proteasas y el efecto de iones bajo la forma de cloruros. Resultados. La enzima mostró homología en la estructura primaria con otras TLEs reportadas para la familia Viperidae, la dosis coagulante mínima (DCM sobre plasma y fibrinógeno humano fue de 18 y 6 µg respectivamente y su potencia coagulante fue de 131,1 NHI unidades de trombina. La enzima se mostró estable a condiciones fisiológicas y prescinde de iones para su actividad. Los carbohidratos asociados detectados fueron hexosas (25,76%, hexosaminas (13,1% y ácido siálico (0,76%. Los agentes fluoruro de fenil metil sulfonil floruro (PMSF ditiotreitol (DTT fueron los principales inhibidores de la actividad enzimática en tanto que la heparina no tuvo efecto inhibidor. Conclusiones. El principio coagulante del veneno de Bothrops pictus es una enzima similar a trombina

  11. Snake population venomics and antivenomics of Bothrops atrox: Paedomorphism along its transamazonian dispersal and implications of geographic venom variability on snakebite management.

    Science.gov (United States)

    Calvete, Juan J; Sanz, Libia; Pérez, Alicia; Borges, Adolfo; Vargas, Alba M; Lomonte, Bruno; Angulo, Yamileth; Gutiérrez, José María; Chalkidis, Hipócrates M; Mourão, Rosa H V; Furtado, M Fatima D; Moura-Da-Silva, Ana M

    2011-04-01

    We describe two geographically differentiated venom phenotypes across the wide distribution range of Bothrops atrox, from the Colombian Magdalena Medio Valley through Puerto Ayacucho and El Paují, in the Venezuelan States of Amazonas and Orinoquia, respectively, and São Bento in the Brazilian State of Maranhão. Colombian and Venezuelan venoms show an ontogenetic toxin profile phenotype whereas Brazilian venoms exhibit paedomorphic phenotypes. Venoms from each of the 16 localities sampled contain both population-specific toxins and proteins shared by neighboring B. atrox populations. Mapping the molecular similarity between conspecific populations onto a physical map of B. atrox range provides clues for tracing dispersal routes that account for the current biogeographic distribution of the species. The proteomic pattern is consistent with a model of southeast and southwest dispersal and allopatric fragmentation northern of the Amazon Basin, and trans-Amazonian expansion through the Andean Corridor and across the Amazon river between Monte Alegre and Santarém. An antivenomic approach applied to assess the efficacy towards B. atrox venoms of two antivenoms raised in Costa Rica and Brazil using Bothrops venoms different than B. atrox in the immunization mixtures showed that both antivenoms immunodepleted very efficiently the major toxins (PIII-SVMPs, serine proteinases, CRISP, LAO) of paedomorphic venoms from Puerto Ayacucho (Venezuelan Amazonia) through São Bento, but had impaired reactivity towards PLA(2) and P-I SVMP molecules abundantly present in ontogenetic venoms. The degree of immunodepletion achieved suggests that each of these antivenoms may be effective against envenomations by paedomorphic, and some ontogenetic, B. atrox venoms. PMID:21278006

  12. Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil Sarcophagidae e Calliphoridae associados às carcaças de Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes e Mabuya frenata (Reptilia, Lacertilia em Brasília, Distrito Federal, Brasil

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    Roger Maia Dias Ledo

    2012-09-01

    Full Text Available Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil. This paper presents a list of necrophagous insects associated with small size carrions of two reptiles and one amphibian, found in areas of riparian forests and Cerrado sensu stricto physiognomies in a Conservation Unit located in Brasilia, Distrito Federal. We found seven species of insects related to these carcasses, being five Sarcophagidae, one Calliphoridae and one Braconidae parasitoid wasp. Lucilia eximia and Peckia (Pattonella intermutans were the most abundant species in the study, corroborating with other studies that suggests that these species have specializations for colonization of small size animal carcasses.Sarcophagidae e Calliphoridae associados às carcaças de Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes e Mabuya frenata (Reptilia, Lacertilia em Brasília, Distrito Federal, Brasil. Este trabalho apresenta uma lista de insetos decompositores associados a carcaças de pequeno porte de dois répteis e de um anfíbio, encontrados em áreas de matas de galeria e de cerrado sensu stricto em unidades de conservação do Distrito Federal. Foram encontradas sete espécies de insetos associados a essas carcaças, sendo cinco sarcofagídeos, um califorídeo e uma vespa parasitóide Braconidae. Lucilia eximia e Peckia (Pattonella intermutans foram as espécies mais abundantes, corroborando com outros estudos que sugerem que estas espécies apresentam especializações para a colonização de carcaças menores.

  13. Comparação entre a epidemiologia do acidente e a clínica do envenenamento por serpentes do gênero Bothrops, em adultos idosos e não idosos Comparison between the epidemiology of accidents and the clinical features of envenoming by snakes of the genus Bothrops, among elderly and non-elderly adults

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    Lindioneza Adriano Ribeiro

    2008-02-01

    Full Text Available O objetivo do presente estudo foi conhecer diferenças epidemiológicas e clínicas do envenenamento por Bothrops spp em adultos idosos (>60 anos e não idosos (20 a 59 anos. Os dados foram obtidos de 1.930 prontuários de pacientes atendidos no Instituto Butantan de 1981 a 1992. Quanto maior a idade do paciente maior a freqüência do acometimento das mãos em relação aos pés (p12 horas após a picada (pThis study had the aim of ascertaining epidemiological and clinical differences in envenoming caused by Bothrops spp between elderly adults (>60 years and non-elderly adults (20 to 59 years. The data were obtained from 1,930 medical records of patients attended at the Butantan Institute between 1981 and 1992. The greater the patient’s age was, the higher the frequency of bites on the hands rather than on the feet was (p 12 hours after the bite (p < 0.05. Necrosis was more common among the elderly patients (p < 0.05 and renal failure was more common among patients aged 50 years or over (p < 0.05, in relation to younger patients. It was concluded that elderly individuals are more often bitten on the hands and less often on the feet, and that they develop local necrosis and renal failure more frequently than do younger individuals.

  14. Alkylation of Histidine Residues of Bothrops jararacussu Venom Proteins and Isolated Phospholipases A2: A Biotechnological Tool to Improve the Production of Antibodies

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    C. L. S. Guimarães

    2014-01-01

    Full Text Available Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2 of this toxin (BthTX-I and BthTX-II were chemically modified (alkylation by p-bromophenacyl bromide (BPB in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated. BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2 diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom.

  15. Purification and partial characterization of phospholipases A2 from Bothrops asper (barba amarilla snake venom from Chiriguaná (Cesar, Colombia

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    J. Ramírez-Avila

    2004-01-01

    Full Text Available Components with phospholipase A2 activity were isolated by gel filtration and cationic exchange chromatography from the venom of Bothrops asper snakes from Chiriguaná, Colombia (9°22´N; 73°37´W. Five fractions were obtained by the gel filtration, and PLA2 activity was found in fraction 3 (F3. In the cationic exchange chromatography, F3 showed eight components with PLA2 activity. Six of these components appeared as one band in polyacrylamide gel electrophoresis (SDS-PAGE. Fractions II and VII exhibited an optimal activity at pH 9 and 52ºC. The optimum calcium concentration for fraction II was 48 mM and for fraction VII, 384 mM. Both fractions showed thermal stability. Fraction II was stable at pH values between 2.5 and 9, and fraction VII, between 2.5 and 8. The Michaelis Menten constant (K M was 3.5x10-3 M for fraction II and 1.6x10-3 M for fraction VII. The molecular weight was 16,000 Dalton for fraction II and 17,000 Dalton for fraction VII. Both isoenzymes did not show any toxic activity (DL50 at 5.3 and 4 µg/g. The two fractions showed different kinetic constant (K M, calcium requirement, and substrate specificity for haemolytic activity.

  16. In Vitro Antiplasmodial Activity of Phospholipases A2 and a Phospholipase Homologue Isolated from the Venom of the Snake Bothrops asper

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    Juan Carlos Alarcón Pérez

    2012-12-01

    Full Text Available The antimicrobial and antiparasite activity of phospholipase A2 (PLA2 from snakes and bees has been extensively explored. We studied the antiplasmodial effect of the whole venom of the snake Bothrops asper and of two fractions purified by ion-exchange chromatography: one containing catalytically-active phospholipases A2 (PLA2 (fraction V and another containing a PLA2 homologue devoid of enzymatic activity (fraction VI. The antiplasmodial effect was assessed on in vitro cultures of Plasmodium falciparum. The whole venom of B. asper, as well as its fractions V and VI, were active against the parasite at 0.13 ± 0.01 µg/mL, 1.42 ± 0.56 µg/mL and 22.89 ± 1.22 µg/mL, respectively. Differences in the cytotoxic activity on peripheral blood mononuclear cells between the whole venom and fractions V and VI were observed, fraction V showing higher toxicity than total venom and fraction VI. Regarding toxicity in mice, the whole venom showed the highest lethal effect in comparison to fractions V and VI. These results suggest that B. asper PLA2 and its homologue have antiplasmodial potential.

  17. Gamma radiation affects the anti-Leishmania activity of Bothrops moojeni venom and correlates with L-amino acid oxidase activity

    International Nuclear Information System (INIS)

    Leishmania causes human disfiguring skin disease in endemic areas of Amazon and North Eastern Brazil. Those parasites present a remarkable resistance to most treatments, except those using toxic antimonial salts. We detected a specific anti-Leishmania activity in snake venoms, using an in vitro promastigote assay. In this report, we analyzed the activity of Bothrops moojeni venom against L. Amazonensis, using whole venom or fractions of L-amino acid oxidase (L-AO). Crude venom of B.moojeni, was fractionated by molecular exclusion chromatography. Activity against promastigotes was detected by respiratory oxidative conversion of MTT in a colorimetric assay and L-AO activity was detected by a colorimetric assay with peroxidase and OPD as revealing reagents. Crude venom was irradiated with 500, 1000, and 2000 Gy in a 60 Co gamma radiation source. The venom had an anti-Leishmania activity of 33 pg/promastigote and the active fraction migrates as 100-150 kDa, close to the size described for L-AOs, and also presented L-AO activity. The radiation reduces both the L-AO and anti-Leishmania activity in a dose dependent effect. Those data suggests the anti-Leishmania activity in this venom is closely related to the L-amino acid oxidase activity and also that radiation could be used as a tool to detect specific activities reduction in water solutions, similarly to observed in dry preparations. (author)

  18. Sexual dimorphism in development and venom production of the insular threatened pit viper Bothrops insularism (Serpentes: Viperidae of Queimada Grande Island, Brazil

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    S.R. Travaglia-Cardoso

    2010-09-01

    Full Text Available Bothrops insularis is a threatened snake endemic to Queimada Grande Island, southern coast of São Paulo, Brazil, and the occurrence of sexual abnormalities in females (females with functional ovaries and rudimentary hemipenis has been reported in this population. To date there are few data regarding developmental features of this particular species. The aim of this study was to follow some developmental features in specimens maintained in captivity for seven years in the Herpetology Laboratory at Instituto Butantan, São Paulo, Brazil. We verified a pronounced sexual dimorphism in development and venom production in the specimens analyzed. In this regard, females showed greater length, mass and amount of venom in comparison to males. Our results suggest a possible niche partitioning between the sexes that reduces (or minimizes intraspecific disharmonic interactions (eg. competition on their small living area (Queimada Grande Island. Taken together, our data suggest that males and females probably are divergent in their diets, with females feeding preferentially on endothermic prey (such as migratory birds, while males maintain the juvenile diet (with the major items being ectothermic prey.

  19. Extracts of Renealmia alpinia (Rottb. MAAS Protect against Lethality and Systemic Hemorrhage Induced by Bothrops asper Venom: Insights from a Model with Extract Administration before Venom Injection

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    Arley Camilo Patiño

    2015-04-01

    Full Text Available Renealmia alpinia (Rottb. MAAS, obtained by micropropagation (in vitro and wild forms have previously been shown to inhibit some toxic activities of Bothrops asper snake venom if preincubated before injection. In this study, assays were performed in a murine model in which extracts were administered for three days before venom injection. R. alpinia extracts inhibited lethal activity of B. asper venom injected by intraperitoneal route. Median Effective Dose (ED50 values were 36.6 ± 3.2 mg/kg and 31.7 ± 5.4 mg/kg (p > 0.05 for R. alpinia wild and in vitro extracts, respectively. At a dose of 75 mg/kg, both extracts totally inhibited the lethal activity of the venom. Moreover, this dose prolonged survival time of mice receiving a lethal dose of venom by the intravenous route. At 75 mg/kg, both extracts of R. alpinia reduced the extent of venom-induced pulmonary hemorrhage by 48.0% (in vitro extract and 34.7% (wild extract, in agreement with histological observations of lung tissue. R. alpinia extracts also inhibited hemorrhage in heart and kidneys, as evidenced by a decrease in mg of hemoglobin/g of organ. These results suggest the possibility of using R. alpinia as a prophylactic agent in snakebite, a hypothesis that needs to be further explored.

  20. Structural and functional characterization of myotoxin I, a Lys49 phospholipase A2 homologue from the venom of the snake Bothrops atrox.

    Science.gov (United States)

    Núñez, Vitelbina; Arce, Viviana; Gutiérrez, José María; Lomonte, Bruno

    2004-07-01

    A new myotoxin was isolated from the venom of Bothrops atrox from Colombia. B. atrox myotoxin I is a homodimer, with a subunit molecular mass of 13,826, and a pI of 8.9. Its complete nucleotide sequence was obtained by cDNA cloning, indicating a mature product of 122 residues that belongs to the family of Lys49 phospholipase A(2) (PLA(2)) homologues, a subgroup of catalytically inactive proteins within the group IIA. Accordingly, the toxin was devoid of phospholipase and anticoagulant activities, in vitro. In mice, it induced conspicuous local myonecrosis, edema, and a systemic interleukin-6 response. In vitro, it was cytolytic upon myoblasts, and weakly bactericidal. The toxin showed highest homology with other Lys49 PLA(2)s, both in its primary and three-dimensional modeled structure, although with an evident difference in the C-terminal region. Unlike Lys49 proteins of American crotalids having 121 residues, this toxin presents an insertion (Asn) between positions 118 and 119. Despite several substitutions within the C-terminal region 115-129 between B. atrox myotoxin I and B. asper myotoxin II, antibodies against synthetic peptide 115-129 of the latter were strongly cross-reactive to the former, indicating the antigenic conservation of this site, known to be critical for the membrane-damaging activities of Lys49 myotoxins. PMID:15225567

  1. Low-molecular-mass peptides from the venom of the Amazonian viper Bothrops atrox protect against brain mitochondrial swelling in rat: potential for neuroprotection.

    Science.gov (United States)

    Martins, N M; Ferreira, D A S; Carvalho Rodrigues, M A; Cintra, A C O; Santos, N A G; Sampaio, S V; Santos, A C

    2010-08-01

    The neurodegenerative diseases are important causes of morbidity and mortality in Western countries. Common mechanisms of toxicity involving mitochondrial damage have been suggested; however, a definitive treatment has not yet been found. Therefore, there has been great interest in the development of mitochondria-targeted protective compounds for the treatment of neuropathies. Animal toxins represent a promising source of new molecules with neuroprotective activity and potential to originate new drugs. We present here the effects of a low-molecular-mass peptides fraction (Ba-V) from Bothrops atrox snake venom, on rat brain mitochondrial function. Ba-V did not induce the mitochondrial swelling and moreover, was as effective as cyclosporin A (CsA) to inhibit the calcium/phosphate-induced swelling, which indicates its potential to prevent the mitochondrial permeability transition (MPT). The membrane electrochemical potential, the oxygen consumption during states-3 and -4 respirations as well as the respiratory control ratio (RCR) were not affected by Ba-V. Additionally, Ba-V did not induce reactive oxygen species (ROS) generation. Interestingly, Ba-V did not protect against the generation of ROS induced by t-BOH, which suggests a protection mechanism other than ROS scavenging. Given the important role of the mitochondrial damage and, more specifically, of MPT, in the development of neuropathies, Ba-V might be useful in the future strategies for the treatment of these diseases. PMID:20338188

  2. The novel metalloproteinase atroxlysin-I from Peruvian Bothrops atrox (Jergón) snake venom acts both on blood vessel ECM and platelets.

    Science.gov (United States)

    Sanchez, Eladio F; Schneider, Francisco S; Yarleque, Armando; Borges, Marcia H; Richardson, Michael; Figueiredo, Suely G; Evangelista, Karla S; Eble, Johannes A

    2010-04-01

    We report the isolation and structure-function relationship of a 23kDa metalloproteinase named atroxlysin-I from the venom of the Peruvian Bothrops atrox (Jergón). Atroxlysin is a P-I metalloproteinase and contains 204 residues. Its proteolytic activity towards dimethylcasein is enhanced by Ca2+ but inhibited by EDTA, dithiothreitol, excessive Zn2+ and alpha2-macroglobulin. Unlike other structurally homologous P-I metalloproteinases, atroxlysin-I causes hemorrhages. To examine its hemorrhagic activity mechanistically, we studied its function in vitro and in vivo. It cleaved the Ala14-Leu15 and Tyr16-Leu17 bonds in oxidized insulin B-chain and specifically hydrolyzed the alpha-chains of fibrin(ogen) in a dose- and time-dependent manner. Atroxlysin-I cleaved plasma fibronectin and other extracellular matrix proteins (collagens I and IV) and the triple-helical fragment CB3 of collagen IV, but did not degrade laminin-111. Complementarily, the laminin and collagen binding integrins alpha7beta1 and alpha1beta1 were cleaved by atroxlysin. Even without catalytic activity atroxlysin-I inhibited collagen- and ADP-triggered platelet aggregation. PMID:20102699

  3. Osteopontin, a chemotactic protein with cytokine-like properties, is up-regulated in muscle injury caused by Bothrops lanceolatus (fer-de-lance) snake venom.

    Science.gov (United States)

    Barbosa-Souza, Valéria; Contin, Daniel Kiss; Filho, Waldemar Bonventi; de Araújo, Albetiza Lôbo; Irazusta, Silvia Pierre; da Cruz-Höfling, Maria Alice

    2011-10-01

    Osteopontin (OPN) is a chemotactic, adhesive protein whose receptors include some integrins and matrix proteins known to have role in inflammatory and repair processes. We examined the time course of OPN expression at acute and chronic stages after intramuscular injection of Bothrops lanceolatus venom in rats. Additionally, we examined the expression of CD68 (a marker for phagocytic macrophages) and the myogenic factors, myoD and myogenin. There was a biphasic upregulation of OPN (6-48 h and 3-14 days post-venom), i.e., during acute inflammation and myogenic cell proliferation and differentiation phases. OPN was detected in CD68 + macrophages, fibroblasts, normal and damaged myofibers, myoblasts and myotubes. Myogenin was expressed in the cytoplasm (atypical pattern) and nucleus of myoblasts and myotubes from 18 h to 7 days, after which it was expressed only in nuclei. Macrophage numbers, OPN and myogenin expression were still elevated at 7, 14 and 7 days. At 3 days, when OPN achieved the peak, some clusters of myoblasts were within regions of intense collagen deposition. Fibrosis may represent limitation for repairing processes and may explain the small diameter of regenerated fibers at 21 days post-venom. The expression of OPN in the course of venom-induced damage and regeneration suggests stages-specific mediation role along the whole process. PMID:21839764

  4. Neutralization, by a monospecific Bothrops lanceolatus antivenom, of toxic activities induced by homologous and heterologous Bothírops snake venoms.

    Science.gov (United States)

    Bogarín, G; Romero, M; Rojas, G; Lutsch, C; Casadamont, M; Lang, J; Otero, R; Gutiérrez, J M

    1999-03-01

    A monospecific Bothrops lanceolatus antivenom, currently used in Martinique, was tested for its efficacy in the neutralization of several toxic and enzymatic activities of the venoms of B. lanceolatus, B. atrox and B. asper. When tested by the i.p. route in mice, B. lanceolatus venom had an LD50 of 12.8 microg/g. In addition, it induced local tissue damage (hemorrhage, edema and myotoxicity) and showed indirect hemolytic activity, but was devoid of coagulant effect on human plasma in vitro and of defibrinating activity in mice. Antivenom was fully effective in the neutralization of lethal, hemorrhagic, edema-forming, myotoxic and indirect hemolytic effects of B. lanceolatus venom in assays involving preincubation of venom and antivenom. When tested against the venoms of B. asper and B. atrox, the antivenom completely neutralized the lethal, hemorrhagic, myotoxic and indirect hemolytic effects, and was partially effective in neutralizing edema-forming activity. In contrast, the antivenom was ineffective in the neutralization of in vitro coagulant and in vivo defibrinating effects induced by these two venoms. PMID:10080358

  5. Quadros clínico-patológicos do envenenamento ofídico por Crotalus durissus terrificus e Bothrops spp. em animais de produção

    Directory of Open Access Journals (Sweden)

    Carlos Hubinger Tokarnia

    2014-04-01

    Full Text Available Foi realizada uma revisão dos quadros clínico-patológicos causados pelos venenos de Crotalus durissus terrificus e Bothrops spp. em bovinos, búfalos, ovinos equinos e suínos. Foram compilados os dados obtidos pela experimentação em animais de produção encontrados na literatura e os obtidos através de experimentação realizada por nossa equipe. Também foram revisados os casos naturais de envenenamento ofídico comunicados. Em dois Quadros foram lançados os mais importantes dados dessas revisões, que revelou diversos aspectos interessantes: 1 em nossos experimentos, o veneno de Crotalus durissus terrificus, quando injetado por via subcutânea em cavalos, causou um edema acentuado no local da aplicação, ao contrário do que tem sido observado em todas as outras espécies animais, aspecto não relatado na literatura; 2 em nossos experimentos, o veneno de diversas espécies de Bothrops, quando injetado por via subcutânea em bovinos, ovinos e equinos, não causou edema como em geral é relatado na literatura, e sim hemorragias subcutâneas acentuadas no local da aplicação. Nos casos não fatais este sangue era reabsorvido em poucos dias sem deixar sequelas. Exceção foi a reação ao veneno de Bothrops jararacussu, que causou edema nos ovinos experimentais, e tumefação acentuada que resultou em fístula com eliminação de líquido seroso nos equinos experimentais. O objetivo do presente estudo visa contribuir para o aperfeiçoamento do diagnóstico de acidentes ofídicos em animais de produção.

  6. Evaluación biológica preliminar de extractos vegetales utilizados en la medicina tradicional de la Sierra Nevada deSanta Marta contra el veneno de la Bothrops asper

    OpenAIRE

    Willinton Barranco Pérez; Vitelbina Nuñez; Mauricio Sanchez

    2013-01-01

    Title: Preliminary biological evaluation of plants extracts used in the Sierra Nevada de Santa Marta against the snake Bothrops asper venom.ResumenLa mordedura de serpientes constituye un problema de salubridad importante en muchos países tropicales y subtropicales, con un estimado de 2,5 millones de personas envenenadas cada año. En Colombia las especies Bothropsasper y Bothropsatrox son las causantes del 70 al 90 % de los accidentes registrados. Se estima que el 60% de estos accidentes son ...

  7. Ineficácia do antiveneno botrópico na neutralização da atividade coagulante do veneno de Lachesis muta muta: relato de caso e comprovação experimental Inefficacy of bothropic antivenom on the neutralization of the coagulant activity of Lachesis muta muta venom: case report and experimental evidence

    Directory of Open Access Journals (Sweden)

    Ramza Bard

    1994-02-01

    Full Text Available No Estado do Amazonas, nas regiões circunvizinhas à cidade de Manaus, as principais espécies de serpentes causadoras de acidentes são Bothrops atrox e Lachesis muta muta com um percentual de ocorrência, dos acidentes confirmados, de 76% e 17%, respectivamente. Rotineiramente, na ausência dos soros antilaquético e antibotrópico-laquético o Instituto de Medicina Tropical de Manaus (IMTM, utiliza-se do soro antibotrópico no tratamento do acidente laquético. Neste trabalho relatamos um caso de acidente por L m. muta, onde o paciente foi tratado com 20 ampolas do soro antibotrópico e permaneceu com o sangue incoagulável até o 13º dia após o acidente. Experimentos foram realizados para obtenção das potências do soro antibotrópico para as atividades coagulante e hemorrágica dos venenos de L. m. muta e de B. atrox. Os resultados mostram que as potências do soro para a atividade hemorrágica dos venenos de L. m. muta e de B. atrox foram similares enquanto que a potência, para a atividade coagulante do veneno de L. m. muta, foi 9,2 vezes menor. Os títulos de anticorpos de três diferentes lotes de soro antibotrópico variaram para o veneno de L. m. muta e foram constantes para o veneno de B. atrox. Devido a ineficácia do soro antibotrópico em neutralizar, principalmente, a atividade coagulante do veneno de L. m. muta, sugerimos a não utilização do antibotrópico no tratamento dos acidentes por L. m. muta.In the State of Amazonas and in areas nearby Manaus, the snakes responsable for the majority of accidents are Bothrops atrox and Lachesis muta muta, with a percentage of confirmed species of 76% and 17%, respectively. Frequently, in the absence of the laquetic and bothropic-laquetic antivenoms, the Instituto de Medicina Tropical de Manaus (IMTM has been using bothropic antivenom in the treatment of laquetic accident. In this paper is related a case of accident caused by L. muta muta; the patient was treated with bothropic

  8. Snake venomics and antivenomics of Bothrops atrox venoms from Colombia and the Amazon regions of Brazil, Perú and Ecuador suggest the occurrence of geographic variation of venom phenotype by a trend towards paedomorphism.

    Science.gov (United States)

    Núñez, Vitelbina; Cid, Pedro; Sanz, Libia; De La Torre, Pilar; Angulo, Yamileth; Lomonte, Bruno; Gutiérrez, José María; Calvete, Juan J

    2009-11-01

    The venom proteomes of Bothrops atrox from Colombia, Brazil, Ecuador, and Perú were characterized using venomic and antivenomic strategies. Our results evidence the existence of two geographically differentiated venom phenotypes. The venom from Colombia comprises at least 26 different proteins belonging to 9 different groups of toxins. PI-metalloproteinases and K49-PLA(2) molecules represent the most abundant toxins. On the other hand, the venoms from Brazilian, Ecuadorian, and Peruvian B. atrox contain predominantly PIII-metalloproteinases. These toxin profiles correlate with the venom phenotypes of adult and juvenile B. asper from Costa Rica, respectively, suggesting that paedomorphism represented a selective trend during the trans-Amazonian southward expansion of B. atrox through the Andean Corridor. The high degree of crossreactivity of a Costa Rican polyvalent (Bothrops asper, Lachesis stenophrys, Crotalus simus) antivenom against B. atrox venoms further evidenced the close evolutionary kinship between B. asper and B. atrox. This antivenom was more efficient immunodepleting proteins from the venoms of B. atrox from Brazil, Ecuador, and Perú than from Colombia. Such behaviour may be rationalized taking into account the lower content of poorly immunogenic toxins, such as PLA(2) molecules and PI-SVMPs in the paedomorphic venoms. The immunological profile of the Costa Rican antivenom strongly suggests the possibility of using this antivenom for the management of snakebites by B. atrox in Colombia and the Amazon regions of Ecuador, Perú and Brazil. PMID:19665598

  9. Anti-parasitic effect on Toxoplasma gondii induced by BnSP-7, a Lys49-phospholipase A2 homologue from Bothrops pauloensis venom.

    Science.gov (United States)

    Borges, Isabela Pacheco; Castanheira, Letícia Eulalio; Barbosa, Bellisa Freitas; de Souza, Dayane Lorena Naves; da Silva, Rafaela José; Mineo, José Roberto; Tudini, Kelly Aparecida Yoneyama; Rodrigues, Renata Santos; Ferro, Eloísa Amália Vieira; de Melo Rodrigues, Veridiana

    2016-09-01

    Toxoplasmosis affects a third of the global population and presents high incidence in tropical areas. Its great relevance in public health has led to a search for new therapeutic approaches. Herein, we report the antiparasitic effects of BnSP-7 toxin, a Lys49 phospholipase A2 (PLA2) homologue from Bothrops pauloensis snake venom, on Toxoplasma gondii. In an MTT assay, BnSP-7 presented significant cytotoxicity against host HeLa cells at higher doses (200 μg/mL to 50 μg/mL), whereas lower doses (25 μg/mL to 1.56 μg/mL) produced low cytotoxicity. Furthermore, the toxin showed no effect on T. gondii tachyzoite viability when evaluated by trypan blue exclusion, but decreased both adhesion and parasite proliferation when tachyzoites were treated before infection. We also measured cytokines in supernatants collected from HeLa cells infected with T. gondii tachyzoites previously treated with RPMI or BnSP-7, which revealed enhancement of only MIF and IL-6 cytokines levels in supernatants of HeLa cells after BnSP-7 treatment. Our results showed that the BnSP-7 PLA2 exerts an anti-Toxoplasma effect at a lower dose than that required to induce cytotoxicity in HeLa cells, and also modulates the immune response of host cells. In this sense, the anti-parasitic effect of BnSP-7 PLA2 demonstrated in the present study opens perspectives for use of this toxin as a tool for future studies on toxoplasmosis. PMID:27212627

  10. Action of Bothrops moojeni venom and its L-amino acid oxidase fraction, treated with 60Co gamma rays, in Leishmania spp

    International Nuclear Information System (INIS)

    Bothrops moojeni venom showed an anti leishmania activity in vitro, as determined by a cell viability assay using the reduction of MTT. After venom purification, by chromatography techniques, the fractions with anti leishmania and L-amino acid oxidase activities, eluted in the same positions. The molecular weight of the enzyme was estimated to be 140 kDa by molecular exclusion chromatography, and 69 kDa, by SDS-PAGE, migrating as a single band, with an isoelectric point of 4.8 as determined by isoelectric focusing. The purified LAO from B. moojeni venom, 135-fold more active than crude venom, showed homo dimeric constitution, and was active against Leishmania spp from the New World, with an effective concentration against L(L). amazonensis of 1.80 μg/ml (EC50), L.(V.) panamensis (0.78 |μg/ml) and L.(L.) chagasi (0.63 (μg/ml). Ultrastructural studies of promastigotes affected by LAO demonstrated cell death, with edema in several organelles such as mitochondria and nuclear membrane, before cell disruption and necrosis. The action of LAO was demonstrated to be hydrogen peroxide-dependent. Studies with LLCMK-2 cells, treated with LAO, showed a toxic effect, with an EC50 of 11|μg/ml. Irradiation of LAO with 60Co gamma rays, did not affect its whole oxidative activity, neither detoxified the enzyme. Amastigotes treated with LAO were not affected by its hydrogen peroxide, otherwise, the exogenous product, killed amastigotes with an EC50 of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  11. Snakebites and ethnobotany in the northwest region of Colombia: Part II: neutralization of lethal and enzymatic effects of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Jiménez, S L; Fonnegra, R; Osorio, R G; García, M E; Díaz, A

    2000-08-01

    Twelve of 74 ethanolic extracts of plants used by traditional healers for snakebites in the northwest region of Colombia, were active against lethal effect of Bothrops atrox venom when they were i.p. injected into mice (18-20 g). After preincubation of sublethal doses of every extract (0.5-4.0 mg/mouse) with 1.5 i.p. lethal dose 50% (LD50) (99.3 microg) of venom, seven of them demonstrated 100% neutralizing capacity within 48 h. These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); rhizomes of Renealmia alpinia (Zingiberaceae) and Heliconia curtispatha (Heliconiaceae); the whole plants of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); and the ripe fruits of Citrus limon (Rutaceae). The other five extracts showing partial neutralization (45-80%; 10-30% survival rate in the control group receiving the venom alone; PBixa orellana (Bixaceae) and Struthanthus orbicularis (Loranthaceae). When the extracts were independently administered per oral or i.p. route 60 min before an i.m. venom injection (204 microg=1.5 i.m. LD50), C. limon, T. elegans, B. orellana and T. rosea extracts had partial and significant neutralizing capacity against B. atrox venom lethal effect. C. limon extract was also partially effective when it was administered either i.v. 15 min before or i.p. 5 min after an i.m. venom injection. Three of the 12 extracts with anti-lethal effect (C. limon, D. croatii and S. acuta) were devoid of antiphospholipase A2 activity, when they were tested against one minimum indirect hemolytic dose of B. atrox venom (2 microg) in agarose-erythrocyte-egg yolk gels. PMID:10940590

  12. Anti-parasitic effect on Toxoplasma gondii induced by BnSP-7, a Lys49-phospholipase A2 homologue from Bothrops pauloensis venom.

    Science.gov (United States)

    Borges, Isabela Pacheco; Castanheira, Letícia Eulalio; Barbosa, Bellisa Freitas; de Souza, Dayane Lorena Naves; da Silva, Rafaela José; Mineo, José Roberto; Tudini, Kelly Aparecida Yoneyama; Rodrigues, Renata Santos; Ferro, Eloísa Amália Vieira; de Melo Rodrigues, Veridiana

    2016-09-01

    Toxoplasmosis affects a third of the global population and presents high incidence in tropical areas. Its great relevance in public health has led to a search for new therapeutic approaches. Herein, we report the antiparasitic effects of BnSP-7 toxin, a Lys49 phospholipase A2 (PLA2) homologue from Bothrops pauloensis snake venom, on Toxoplasma gondii. In an MTT assay, BnSP-7 presented significant cytotoxicity against host HeLa cells at higher doses (200 μg/mL to 50 μg/mL), whereas lower doses (25 μg/mL to 1.56 μg/mL) produced low cytotoxicity. Furthermore, the toxin showed no effect on T. gondii tachyzoite viability when evaluated by trypan blue exclusion, but decreased both adhesion and parasite proliferation when tachyzoites were treated before infection. We also measured cytokines in supernatants collected from HeLa cells infected with T. gondii tachyzoites previously treated with RPMI or BnSP-7, which revealed enhancement of only MIF and IL-6 cytokines levels in supernatants of HeLa cells after BnSP-7 treatment. Our results showed that the BnSP-7 PLA2 exerts an anti-Toxoplasma effect at a lower dose than that required to induce cytotoxicity in HeLa cells, and also modulates the immune response of host cells. In this sense, the anti-parasitic effect of BnSP-7 PLA2 demonstrated in the present study opens perspectives for use of this toxin as a tool for future studies on toxoplasmosis.

  13. Comparison of venoms from wild and long-term captive Bothrops atrox snakes and characterization of Batroxrhagin, the predominant class PIII metalloproteinase from the venom of this species.

    Science.gov (United States)

    Freitas-de-Sousa, L A; Amazonas, D R; Sousa, L F; Sant'Anna, S S; Nishiyama, M Y; Serrano, S M T; Junqueira-de-Azevedo, I L M; Chalkidis, H M; Moura-da-Silva, A M; Mourão, R H V

    2015-11-01

    Comparisons between venoms from snakes kept under captivity or collected at the natural environment are of fundamental importance in order to obtain effective antivenoms to treat human victims of snakebites. In this study, we compared composition and biological activities of Bothrops atrox venom from snakes collected at Tapajós National Forest (Pará State, Brazil) or maintained for more than 10 years under captivity at Instituto Butantan herpetarium after have been collected mostly at Maranhão State, Brazil. Venoms from captive or wild snakes were similar except for small quantitative differences detected in peaks correspondent to phospholipases A2 (PLA2), snake venom metalloproteinases (SVMP) class PI and serine proteinases (SVSP), which did not correlate with fibrinolytic and coagulant activities (induced by PI-SVMPs and SVSPs). In both pools, the major toxic component corresponded to PIII-SVMPs, which were isolated and characterized. The characterization by mass spectrometry of both samples identified peptides that matched with a single PIII-SVMP cDNA characterized by transcriptomics, named Batroxrhagin. Sequence alignments show a strong similarity between Batroxrhagin and Jararhagin (96%). Batroxrhagin samples isolated from venoms of wild or captive snakes were not pro-coagulant, but inhibited collagen-induced platelet-aggregation, and induced hemorrhage and fibrin lysis with similar doses. Results suggest that in spite of environmental differences, venom variability was detected only among the less abundant components. In opposition, the most abundant toxin, which is a PIII-SVMP related to the key effects of the venom, is structurally conserved in the venoms. This observation is relevant for explaining the efficacy of antivenoms produced with venoms from captive snakes in human accidents inflicted at distinct natural environments.

  14. Neutralizing activities of ethanolic extracts of six plants traditionally used in Guatemala as antidotes for the envenomation caused by the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    Patricia Saravia-Otten

    2015-07-01

    Full Text Available Many plants are reported to be used in Guatemalan traditional medicine as antidotes against various effects of the snakebite; however, very few attempts have been made to evaluate their neutralizing capacity in controlled experiments. Six plants (Acacia hindsii, Cissampelos pareira; Hamelia patens, Piper peltatum, Sansevieria hyacinthoides and Aristolochia maxima were evaluated in vitro for their ability to neutralize phospholipase A2(PLA2 and proteolytic effects of the venom of Bothrops asper, the snake responsible for approximately half of the snakebite envenomations in Central America. These effects are indicatives of the ability of B. asper venom to produce myotoxicity, hemorrhage and inflammation. Plants were collected, dried and extracted by maceration with ethanol. After pre-incubation of several amounts of each extract with a challenge dose of venom, S. hyacinthoides demonstrated a low neutralizing capacity (< DE 50 of the PLA2 effect (13.90 ± 6.41%; C. pareira (32.98 ± 5.51% and P. peltatum (24.52 ± 7.45% neutralized less than 50% of the proteolytic effect. The results suggest that neither of the tested plants should be used individually to treat the main effects of B. asper envenomation. However, the three low-active extracts might be potentiated when used in mixtures composed of several plants, as prepared by traditional healers. Given the complexity of the venom components and the multiple pathologic effects produced by B. asper envenomation, more tests are required to fully investigate the ability of this plants to neutralize the coagulant, fibrin(ogenolytic, edematizing and myotoxic effects of the venom.

  15. Isolation and characterization of a new serine protease with thrombin-like activity (TLBm) from the venom of the snake Bothrops marajoensis.

    Science.gov (United States)

    Vilca-Quispe, Augusto; Ponce-Soto, Luis Alberto; Winck, Flavia Vischi; Marangoni, Sergio

    2010-04-01

    The thrombin-like serine protease TLBm from Bothrops marajoensis was isolated in one chromatographic step in reverse phase HPLC. Its molecular mass was 33239.95 Da, as based on the determined primary structure and confirmed experimentally by MALDI-TOF mass spectrometry (33332.5 Da) and it contains 12 half-cysteine residues. This TLBm exhibited high specificity for BArhoNA, Michaelis-Menten behavior with K(m) 2.3x10(-1)M and the V(max) 0.52x10(-1) nmoles rho-NA/lt/min for this substrate. TLBm also showed ability to coagulate bovine fibrinogen and was inhibited by soybean trypsin inhibitor, EDTA and S(Dm) from the serum of the species Didelphis marsupialis. The primary structure of TLBm showed the presence of His(45), Asp(103) and Ser(228) residues in the corresponding positions of the catalytic triad established in the serine proteases and Ser(228) are inhibited by phenylmethylsulfonyl fluoride (PMSF). Amino acid analysis showed a high content of Asp, Glu, Gly, Ser, Ala and Pro as well as 12 half-cysteine residues and calculated pI of 6.47; TLBm presented 285 amino acid residues. In this work, we investigated the ability of TLBm to degrade fibrinogen and we observed that it is able to cause alpha- and beta-chain cleavage. Enzymatic as well as the platelet aggregation activities were strongly inhibited when incubated with PMSF, a specific inhibitor of serine protease. Also, TLBm induced platelet aggregation in washed and platelet-rich plasma, and in both cases, PMSF inhibited its activity. PMID:19931298

  16. Actividad in vitro de los venenos de Lachesis muta y Bothrops atrox sobre la viabilidad y desarrollo embrionario de los huevos de Ascaris suum

    Directory of Open Access Journals (Sweden)

    Claudia Paredes

    2014-06-01

    Full Text Available Los venenos de serpientes son concentrados polienzimáticos cuya actividad biológica sobre algunas bacterias y protozoos ha sido comprobada. El objetivo principal del presente trabajo fue estudiar la actividad in vitro de los venenos totales de las serpientes Lachesis muta y Bothrops atrox sobre la viabilidad y el desarrollo embrionario de los huevos de Ascaris suum. Se emplearon los venenos totales en concentraciones de 2, 4, 8 y 16 mg/mL sobre huevos no embrionados y larvados in vitro. Se comparó la actividad de los venenos con la de otras sustancias como el hipoclorito de sodio al 5,25%, Albendazol (solución comercial y solución salina. Ambos venenos, en concentraciones de 4, 8 y 16 mg/mL, inhibieron la blastulación de estos huevos; hasta el sexto día de incubación; en cambio en concentración de 2 mg/mL la inhibición se dio hasta el cuarto día. Posteriormente iniciaron un proceso de embrionación aparentemente normal hasta la formación del estadio infectante. El veneno de B. atrox fue el que presentó el mayor efecto inhibitorio en concentración de 16 mg/mL. El hipoclorito de sodio destruyó el 100% de los huevos, mientras el albendazol ocasionó que los huevos iniciaran un proceso de segmentación anormal que originó su degeneración. Se concluye que los venenos de L. muta y B. atrox muestran actividad inhibitoria al inicio de la blastulación de los huevos de A. suum y no ejercen ningún efecto en los huevos larvados.

  17. Prevention of thromboses in human patients with Bothrops lanceolatus envenoming in Martinique: failure of anticoagulants and efficacy of a monospecific antivenom. Research Group on Snake Bites in Martinique.

    Science.gov (United States)

    Thomas, L; Tyburn, B; Bucher, B; Pecout, F; Ketterle, J; Rieux, D; Smadja, D; Garnier, D; Plumelle, Y

    1995-05-01

    Envenomation by the Bothrops lanceolatus, a snake found only in Martinique, leads to swelling and pain, and occasionally to systemic signs and/or coagulopathy. Severe thromboses at some distance from the site of the bite may appear within 48 hr. Uncertainties as to the actual development of thrombotic complications in patients appearing to be suffering from moderate poisoning and as to the availability and the toxicity of a monospecific antivenom (AVS) initially led us to reserve antivenom for the most severe cases, and to use anticoagulants to prevent thromboses in all patients. This approach was modified after we observed serious thromboses in patients with moderate poisoning. Of 50 adult snake bite cases hospitalized between June 1991 and August 1994, 11 developed serious thrombotic complications at 36 /+- 27 hr (mean +/- SD) (range 12-96) following envenomation, despite early preventive anticoagulant therapy. Those included pulmonary embolism (two cases), cerebral infarction (six cases), myocardial infarction (one case), and cerebral and myocardial infarctions (two cases). Sixteen patients were not treated with AVS: 10 of these recovered without complications and six developed systemic thrombosis causing permanent disability in three cases. Thirty were treated with an intravenous infusion of 2-6 vials of AVS given 2-48 hr after the bite. Of these, three died of cerebral infarction that developed before the initiation of serotherapy. All others recovered. Among patients treated with AVS, three presented with mild anaphylactic reactions, while one developed serum sickness that responded to steroids. These data indicate that preventive anticoagulant therapy is of limited efficacy in Martinique.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7771608

  18. Snakebites and ethnobotany in the northwest region of Colombia: Part II: neutralization of lethal and enzymatic effects of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Jiménez, S L; Fonnegra, R; Osorio, R G; García, M E; Díaz, A

    2000-08-01

    Twelve of 74 ethanolic extracts of plants used by traditional healers for snakebites in the northwest region of Colombia, were active against lethal effect of Bothrops atrox venom when they were i.p. injected into mice (18-20 g). After preincubation of sublethal doses of every extract (0.5-4.0 mg/mouse) with 1.5 i.p. lethal dose 50% (LD50) (99.3 microg) of venom, seven of them demonstrated 100% neutralizing capacity within 48 h. These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); rhizomes of Renealmia alpinia (Zingiberaceae) and Heliconia curtispatha (Heliconiaceae); the whole plants of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); and the ripe fruits of Citrus limon (Rutaceae). The other five extracts showing partial neutralization (45-80%; 10-30% survival rate in the control group receiving the venom alone; P<0.05) were: leaves, branches and stem of Costus lasius (Costaceae); the whole plant of Sida acuta (Malvaceae); rhizomes of Dracontium croatii (Araceae); leaves and branches of Bixa orellana (Bixaceae) and Struthanthus orbicularis (Loranthaceae). When the extracts were independently administered per oral or i.p. route 60 min before an i.m. venom injection (204 microg=1.5 i.m. LD50), C. limon, T. elegans, B. orellana and T. rosea extracts had partial and significant neutralizing capacity against B. atrox venom lethal effect. C. limon extract was also partially effective when it was administered either i.v. 15 min before or i.p. 5 min after an i.m. venom injection. Three of the 12 extracts with anti-lethal effect (C. limon, D. croatii and S. acuta) were devoid of antiphospholipase A2 activity, when they were tested against one minimum indirect hemolytic dose of B. atrox venom (2 microg) in agarose-erythrocyte-egg yolk gels.

  19. A comparative analysis of the clotting and fibrinolytic activities of the snake venom (Bothrops atrox) from different geographical areas in Venezuela.

    Science.gov (United States)

    Salazar, Ana Maria; Rodriguez-Acosta, Alexis; Girón, Maria E; Aguilar, Irma; Guerrero, Belsy

    2007-01-01

    Venom constitution within the same snake species can present considerable geographical variations. Bothrops atrox venoms were obtained from adult snakes captured at different geographical locations: Parguasa (Bolívar state); Puerto Ayacucho 1, Serranía del Cuao and Puerto Ayacucho 2 (Amazon state). The coagulant and fibrinolytic activities of these venoms were compared. Amidolytic activity of crude snake venom was measured by a micromethod designed in our laboratory. Coagulant activity on plasma and fibrinogen due to thrombin-like activity in venoms was also determined. Crude snake venom fibrinolytic activity by the fibrin plate method was assayed. Chromatographic studies were developed on Protein-Pack 300 column. Polyacrylamide gel electrophoresis was carried out under reduced conditions. After SDS-PAGE of samples, the fibrin-zymography was tested on agarose-fibrin plates. The results demonstrated several differences among B. atrox venoms from different geographical areas. Chromatograms and SDS-PAGE profiles indicated that venoms from the same species presented differences in the molecular mass of their components. The procoagulant activity depended on the utilized method (amidolytic versus clotting). Parguasa and Puerto Ayacucho 2 venoms presented procoagulant activity for both methods. Furthermore, Parguasa venom had also the highest hemorrhagic activity and the lowest LD50. In relation to the fibrinolytic activity, Puerto Ayacucho 1 venom was the most active, equally for fibrin plates as for the amidolytic method (t-PA like). This venom had the lowest coagulant activity, which induced us to think that probably its procoagulant activity was interfered by its fibrinolytic activity.

  20. Comparison of venoms from wild and long-term captive Bothrops atrox snakes and characterization of Batroxrhagin, the predominant class PIII metalloproteinase from the venom of this species.

    Science.gov (United States)

    Freitas-de-Sousa, L A; Amazonas, D R; Sousa, L F; Sant'Anna, S S; Nishiyama, M Y; Serrano, S M T; Junqueira-de-Azevedo, I L M; Chalkidis, H M; Moura-da-Silva, A M; Mourão, R H V

    2015-11-01

    Comparisons between venoms from snakes kept under captivity or collected at the natural environment are of fundamental importance in order to obtain effective antivenoms to treat human victims of snakebites. In this study, we compared composition and biological activities of Bothrops atrox venom from snakes collected at Tapajós National Forest (Pará State, Brazil) or maintained for more than 10 years under captivity at Instituto Butantan herpetarium after have been collected mostly at Maranhão State, Brazil. Venoms from captive or wild snakes were similar except for small quantitative differences detected in peaks correspondent to phospholipases A2 (PLA2), snake venom metalloproteinases (SVMP) class PI and serine proteinases (SVSP), which did not correlate with fibrinolytic and coagulant activities (induced by PI-SVMPs and SVSPs). In both pools, the major toxic component corresponded to PIII-SVMPs, which were isolated and characterized. The characterization by mass spectrometry of both samples identified peptides that matched with a single PIII-SVMP cDNA characterized by transcriptomics, named Batroxrhagin. Sequence alignments show a strong similarity between Batroxrhagin and Jararhagin (96%). Batroxrhagin samples isolated from venoms of wild or captive snakes were not pro-coagulant, but inhibited collagen-induced platelet-aggregation, and induced hemorrhage and fibrin lysis with similar doses. Results suggest that in spite of environmental differences, venom variability was detected only among the less abundant components. In opposition, the most abundant toxin, which is a PIII-SVMP related to the key effects of the venom, is structurally conserved in the venoms. This observation is relevant for explaining the efficacy of antivenoms produced with venoms from captive snakes in human accidents inflicted at distinct natural environments. PMID:26276061

  1. A comparative analysis of the clotting and fibrinolytic activities of the snake venom (Bothrops atrox) from different geographical areas in Venezuela.

    Science.gov (United States)

    Salazar, Ana Maria; Rodriguez-Acosta, Alexis; Girón, Maria E; Aguilar, Irma; Guerrero, Belsy

    2007-01-01

    Venom constitution within the same snake species can present considerable geographical variations. Bothrops atrox venoms were obtained from adult snakes captured at different geographical locations: Parguasa (Bolívar state); Puerto Ayacucho 1, Serranía del Cuao and Puerto Ayacucho 2 (Amazon state). The coagulant and fibrinolytic activities of these venoms were compared. Amidolytic activity of crude snake venom was measured by a micromethod designed in our laboratory. Coagulant activity on plasma and fibrinogen due to thrombin-like activity in venoms was also determined. Crude snake venom fibrinolytic activity by the fibrin plate method was assayed. Chromatographic studies were developed on Protein-Pack 300 column. Polyacrylamide gel electrophoresis was carried out under reduced conditions. After SDS-PAGE of samples, the fibrin-zymography was tested on agarose-fibrin plates. The results demonstrated several differences among B. atrox venoms from different geographical areas. Chromatograms and SDS-PAGE profiles indicated that venoms from the same species presented differences in the molecular mass of their components. The procoagulant activity depended on the utilized method (amidolytic versus clotting). Parguasa and Puerto Ayacucho 2 venoms presented procoagulant activity for both methods. Furthermore, Parguasa venom had also the highest hemorrhagic activity and the lowest LD50. In relation to the fibrinolytic activity, Puerto Ayacucho 1 venom was the most active, equally for fibrin plates as for the amidolytic method (t-PA like). This venom had the lowest coagulant activity, which induced us to think that probably its procoagulant activity was interfered by its fibrinolytic activity. PMID:17045631

  2. Galatrox is a C-type lectin in Bothrops atrox snake venom that selectively binds LacNAc-terminated glycans and can induce acute inflammation.

    Science.gov (United States)

    Sartim, Marco A; Riul, Thalita B; Del Cistia-Andrade, Camillo; Stowell, Sean R; Arthur, Connie M; Sorgi, Carlos A; Faccioli, Lucia H; Cummings, Richard D; Dias-Baruffi, Marcelo; Sampaio, Suely V

    2014-11-01

    Previous studies indicate that snake venom contains glycan-binding proteins (GBPs), although the binding specificity and biological activities of many of these GBPs is unclear. Here we report our studies on the glycan binding specificity and activities of galatrox, a Bothrops atrox snake venom-derived GBP. Glycan microarray analysis indicates that galatrox binds most strongly to glycans expressing N-acetyllactosamine (LacNAc), with a significant preference for Galβ1-4GlcNAcβ over Galβ1-3GlcNAcβ compounds. Galatrox also bound immobilized laminin, a LacNAc-dense extracellular matrix component, suggesting that this GBP can bind LacNAc-bearing glycoproteins. As several endogenous mammalian GBPs utilize a similar binding LacNAc binding preference to regulate neutrophil and monocyte activity, we hypothesized that galatrox may mediate B. atrox toxicity through regulation of leukocyte activity. Indeed, galatrox bound neutrophils and promoted leukocyte chemotaxis in a carbohydrate-dependent manner. Similarly, galatrox administration into the mouse peritoneal cavity induced significant neutrophil migration and the release of pro-inflammatory cytokines IL-1α and IL-6. Exposure of bone marrow-derived macrophages to galatrox induced generation of pro-inflammatory mediators IL-6, TNF-α, and keratinocyte-derived chemokine. This signaling by galatrox was mediated via its carbohydrate recognition domain by activation of the TLR4-mediated MyD88-dependent signaling pathway. These results indicate that galatrox has pro-inflammatory activity through its interaction with LacNAc-bearing glycans on neutrophils, macrophages and extracellular matrix proteins and induce the release of pro-inflammatory mediators. PMID:24973254

  3. The complement system is involved in acute inflammation but not in the hemorrhage produced by a Bothrops atrox snake venom low molecular mass proteinase.

    Science.gov (United States)

    Rodrigues, Flávia G; Petretski, Jorge H; Kanashiro, Milton M; Lemos, Luciana; da Silva, Wilmar Dias; Kipnis, Thereza L

    2004-03-01

    Low molecular weight hemorrhagins were purified from crude Bothrops atrox snake venom by gel filtration followed by ionic strength chromatography. The protein fractions obtained, designated HI-1 to HI-8, contained proteins with molecular masses lower than 30 kDa. HI-5, the most representative among of these fractions, exhibited, in vitro, proteolytic and C inactivating properties, as analyzed by proteolysis of a protein substrate, and C system consumptive activities as assayed by reduction of the hemolytic C activity in normal human serum and by cleavage of partially purified component C3. HI-5 hemorrhagin injected i.m. into C-sufficient BALB/c mice induced a local inflammation characterized by edema, accumulation of polymorphonuclear leucocytes (PMN) and hemorrhage. In contrast, when injected into BALB/c mice previously C-depleted, the number of PMN per tissue section, but not hemorrhage, was significantly reduced (129.668 +/- 31.341 cells per microscopic field) as compared with the control C-sufficient mice (812.168 +/- 111.194 cells per microscopic field). The observations were confirmed by using C5-deficient mice instead of C-depleted mice. The average number of PMN per tissue section in C5-defficient A/J mice was 72.666 +/- 19.416 cells per microscopic field. These data indicate that the C system is involved in PMN accumulation, but not in the hemorrhage, at the local induced lesions by low molecular mass B. atrox hemorrhagins. HI-5 apparently is not contaminated with other direct or indirect inflammation mediators, PMN accumulation and hemorrhage, however, an independent phenomenon, could be mediated by the same hemorrhagin proteinase domain. PMID:15104120

  4. Use of a synthetic biosensor for neutralizing activity-biased selection of monoclonal antibodies against atroxlysin-I, an hemorrhagic metalloproteinase from Bothrops atrox snake venom.

    Directory of Open Access Journals (Sweden)

    Francisco Santos Schneider

    2014-04-01

    Full Text Available BACKGROUND: The snake Bothrops atrox is responsible for the majority of envenomings in the northern region of South America. Severe local effects, including hemorrhage, which are mainly caused by snake venom metalloproteinases (SVMPs, are not fully neutralized by conventional serum therapy. Little is known about the immunochemistry of the P-I SVMPs since few monoclonal antibodies (mAbs against these molecules have been obtained. In addition, producing toxin-neutralizing mAbs remains very challenging. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report on the set-up of a functional screening based on a synthetic peptide used as a biosensor to select neutralizing mAbs against SVMPs and the successful production of neutralizing mAbs against Atroxlysin-I (Atr-I, a P-I SVMP from B. atrox. Hybridomas producing supernatants with inhibitory effect against the proteolytic activity of Atr-I towards the FRET peptide Abz-LVEALYQ-EDDnp were selected. Six IgG1 Mabs were obtained (named mAbatr1 to mAbatr6 and also two IgM. mAbatrs1, 2, 3 and 6 were purified. All showed a high specific reactivity, recognizing only Atr-I and B. atrox venom in ELISA and a high affinity, showing equilibrium constants in the nM range for Atr-I. These mAbatrs were not able to bind to Atr-I overlapping peptides, suggesting that they recognize conformational epitopes. CONCLUSIONS/SIGNIFICANCE: For the first time a functional screening based on a synthetic biosensor was successfully used for the selection of neutralizing mAbs against SVMPs.

  5. Identificación molecular y actividad sobre sustratos cromogénicos de la venombina A del veneno de la serpiente peruana Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Gustavo A. Sandoval

    2011-05-01

    Full Text Available En el presente trabajo se ha realizado la identificación molecular de la enzima similar a trombina (EST del veneno de Bothrops atrox y se ha evaluado su actividad enzimática sobre diversos sustratos sintéticos. La enzima fue purificada utilizando tres pasos cromatrográficos, sobre Sephadex G-75, CM-Sephadex C-50 y Agarosa-PAB, determinándose su peso molecular por PAGE-SDS. La identificación molecular de la enzima aislada se realizó por la técnica de peptide mass fingerprinting basada en espectrometría de masas MALDI-TOF y posterior análisis in silico. Las actividades fibrinocoagulante y amidolítica fueron ensayadas sobre fibrinó- geno bovino y BApNA, respectivamente, así como la hidrólisis sobre los sustratos cromogénicos específicos S-2238, S-2251 y S-2266. Como resultado de los ensayos bioquímicos y estructurales, la EST del veneno de B. atrox, presentó un peso molecular de 29,6 kDa. El análisis mediante espectrometría de masas de los péptidos obtenidos, permitió identificar a esta enzima como una venombina A, presentando una identidad del 75%. Del análisis de actividad enzimática, se obtuvo que la EST de B. atrox produjo coagulación del fibrinógeno bovino y presentó actividad sobre BApNA, S-2238 y S-2266, siendo incapaz de hidrolizar el sustrato S-2251. El empleo de estas aproximaciones estructurales y funcionales ha permitido lograr la identificación molecular del principal componente del veneno de B. atrox relacionado con su acción coagulante, así como evaluar en detalle la naturaleza de su actividad enzimática sobre diversos sustratos.

  6. Neutrophils regulate the expression of cytokines, chemokines and nitric oxide synthase/nitric oxide in mice injected with Bothrops atrox venom.

    Science.gov (United States)

    Escocard, Rita de Cássia Mothé; Kanashiro, Milton Masahiko; Petretski, Jorge Hudson; Azevedo-Silva, Juliana; Queiroz de Carvalho, Eulógio Carlos; Dias da Silva, Wilmar; Kipnis, Thereza Liberman

    2006-01-01

    Bothrops atrox crude venom injected intraperitoneal (i.p.) into BALB/c mice induced local afflux of inflammatory cells, one neutrophil-rich peak after 6h and another macrophage-rich peak after 48 h. A similar pattern of local cell afflux plus edema, Delta lesions of some skeletal muscle cells, and hemorrhage were observed in mice intramuscular (i.m.) injected with the venom. Measurement of serum cytokines in neutrophil-depleted (by anti-mouse rat monoclonal antibody (mAb) RB6-8C5) and non-depleted BALB/c mice was performed by ELISA. With the exception of IL-1beta (78 pg/ml), higher levels of IL-6 (1348 pg/ml), MIP-1beta (437 pg/ml) and MIP-2 (904 pg/ml) were observed in neutrophil-depleted mice, in comparison to the values found in non-neutrophil depleted mice: IL-1beta (437 pg/ml), IL-6 (750 pg/ml), MIP-1beta (165 pg/ml) and MIP-2 (90 pg/ml). TNF-alpha was not detected. NO was detected (18 microM) 24h after venom injection in neutrophil-depleted mice. RT-PCR using representative primers detected expression of mRNA in cells from BALB/c mice injected with B. atrox venom: (a) for IL-1beta, IL-6, inducible nitric oxide synthase (iNOS), CXCR2, MIP-2 and RANTES in cells from mice that were neutrophil-depleted or not; (b) for CCR1, CCR5 and MIP-1beta in cells from neutrophil-depleted mice; (c) for MIP-1alpha in cells from non-neutrophil-depleted mice; (d) TNF-alpha and TGF-beta were not detected in either of the mice. These results indicate that neutrophils play a role in regulating the production of some cytokines and chemokines as well as locally expressed or liberated iNOS/NO in tissues injected with B. atrox crude venom. PMID:16446169

  7. Evaluación biológica preliminar de extractos vegetales utilizados en la medicina tradicional de la Sierra Nevada deSanta Marta contra el veneno de la Bothrops asper

    Directory of Open Access Journals (Sweden)

    Willinton Barranco Pérez

    2013-10-01

    Full Text Available Title: Preliminary biological evaluation of plants extracts used in the Sierra Nevada de Santa Marta against the snake Bothrops asper venom.ResumenLa mordedura de serpientes constituye un problema de salubridad importante en muchos países tropicales y subtropicales, con un estimado de 2,5 millones de personas envenenadas cada año. En Colombia las especies Bothropsasper y Bothropsatrox son las causantes del 70 al 90 % de los accidentes registrados. Se estima que el 60% de estos accidentes son inicialmente tratados por curanderos tradicionales utilizando plantas medicinales en diferentes preparaciones. Este estudio evaluó la capacidad inhibitoria de cinco especies contra el efecto proteolítico y hemolítico indirecto inducido por el veneno de B. asper en ensayos in vitro.Las especies, que fueron seleccionadas de acuerdo a su uso en la medicina tradicional por parte de las comunidades campesinas de la Sierra Nevada de Santa Marta, fueron, Aristolochia máxima, Cissampelospareira, Equisetumbogotense, Mucunacfpruriens y una especie de la familia Asteraceae. La planta E. bogotense mostró los mayores porcentaje de inhibición contra la actividad de las fosfolipasas A2(42,29 %, así como la mayor precipitación de las proteínas en un rango de masas moleculares de 28,2 y 94,43 KDa. Al fraccionar el extracto de E. bogotense se obtuvieron cinco fracciones, las cuales presentaron un porcentaje de inhibición de 36,6 ± 1,07 a 46,1 ± 13,6. Adicionalmente se detectaron por métodos cualitativos núcleos como, alcaloides, esteroides y/o triterpenos, taninos, cumarinas y leucoantocianidinas. En estudio se reporta la actividad antiofídica en ensayos in vitro de la especie E. bogotense contra el veneno de la especie B.asper. (DUAZARY 2012 No. 2, 140 - 150AbstractIn Colombia the species Bothrops asper and Bothrops atrox are responsible for 70 to 90% of the snakebite accidents. Around 60% of these injuries are initially treated by traditional healers; they

  8. Action of Bothrops moojeni venom and its L-amino acid oxidase fraction, treated with {sup 60}Co gamma rays, in Leishmania spp; Acao do veneno de Bothrops moojeni e sua fracao L-aminoacido oxidase, submetida ao tratamento com raios gama de {sup 60}Co, em Leishmania spp

    Energy Technology Data Exchange (ETDEWEB)

    Cardoso, Andre Gustavo Tempone

    1999-07-01

    Bothrops moojeni venom showed an anti leishmania activity in vitro, as determined by a cell viability assay using the reduction of MTT. After venom purification, by chromatography techniques, the fractions with anti leishmania and L-amino acid oxidase activities, eluted in the same positions. The molecular weight of the enzyme was estimated to be 140 kDa by molecular exclusion chromatography, and 69 kDa, by SDS-PAGE, migrating as a single band, with an isoelectric point of 4.8 as determined by isoelectric focusing. The purified LAO from B. moojeni venom, 135-fold more active than crude venom, showed homo dimeric constitution, and was active against Leishmania spp from the New World, with an effective concentration against L(L). amazonensis of 1.80 {mu}g/ml (EC{sub 50}), L.(V.) panamensis (0.78 |{mu}g/ml) and L.(L.) chagasi (0.63 ({mu}g/ml). Ultrastructural studies of promastigotes affected by LAO demonstrated cell death, with edema in several organelles such as mitochondria and nuclear membrane, before cell disruption and necrosis. The action of LAO was demonstrated to be hydrogen peroxide-dependent. Studies with LLCMK-2 cells, treated with LAO, showed a toxic effect, with an EC{sub 50} of 11|{mu}g/ml. Irradiation of LAO with 6{sup 0C}o gamma rays, did not affect its whole oxidative activity, neither detoxified the enzyme. Amastigotes treated with LAO were not affected by its hydrogen peroxide, otherwise, the exogenous product, killed amastigotes with an EC{sub 50} of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  9. Identificación de Bacterias en Cavidad Oral y Valores Hematológicos en Bothrops asper en el Serpentario de la Universidad de Antioquia -resumen-

    Directory of Open Access Journals (Sweden)

    C Martínez-Mejía

    2014-12-01

    Full Text Available La flora bacteriana de la cavidad oral de las serpientes en cautiverio ha sido asociada a infecciones por estomatitis y abscesos secundarios por auto-mordedura. Igualmente‚ su identificación es importante debido a las infecciones secundarias que pueden causar los accidentes ofídicos en humanos y animales. En particular el veneno de la serpiente mapaná‚ Bothrops asper‚ responsable del 70% de los accidentes ofídicos en Colombia‚ genera efectos sistémicos y locales incluyendo dermonecrosis y mionecrosis que puede complicarse con abscesos‚ celulitis y fascitis hasta en el 30% de los casos. En este estudio se realizaron cultivos bacteriológicos y antibiogramas para identificar las bacterias presentes en la cavidad oral‚ y se determinaron los valores hematológicos no reportados aún para B. asper‚ mantenidas en el serpentario de la Universidad de Antioquia. Mediante hisopados de cavidad oral (n=16 se aislaron 9 especies de bacterias pertenecientes a 4 familias: Enterobacteriaceae (Escherichia coli‚ Citrobacter sp.‚ Proteus sp.‚ Salmonella sp. y Enterobacter sp.‚ Enterococcaceae (Enterococcus sp.‚ Staphylococcaceae (Staphylococcus epidermidis‚ Staphylococcus haemolyticus y Bacillaceae (Bacillus licheniformis. De este total‚ la mayoría (55,5% resultaron Gram negativas y el resto (45,5% fueron Gram positivas. Se observó además una asociación entre los individuos con más tiempo en cautiverio (+ 60 meses‚ 4 ♀ y 3♂ y cultivos simultáneos de 2 bacterias‚ mientras que de los individuos con menos tiempo (- 50 meses‚ 4 ♀ y 5♂ se aisló solo una especie de bacteria. En acuerdo con la literatura‚ todas las bacterias aisladas fueron sensibles a la Amikacina y Enrofloxacina‚ mientras que el 66‚6% (n=9 de las bacterias fue resistente a la Ampicilina Sulbactam. Los parámetros hematológicos se determinaron a partir de 24 serpientes en buen estado de salud. El promedio del hematocrito (Hto‚ % fue de 27

  10. Price increase

    CERN Multimedia

    2006-01-01

    Please take note that after five years of stable prices at Restaurant No 1 a price increase will come into force on 1st January 2006. This increase has been agreed after discussions between the CSR (Comité de Surveillance des Restaurants) and the catering company Novae and will reflect the inflation rate of the last few years. In addition, a new children's menu will be introduced, as well as 'Max Havelaar' fair-trade coffee at a price of 1.70 CHF.

  11. Price increase

    CERN Multimedia

    2005-01-01

    Please take note that after five years of stable prices at Restaurant No 1 a price increase will come into force on 1st January 2006. This increase has been agreed after discussions between the CSR (Comité de Surveillance des Restaurants) and the catering company Novae and will reflect the inflation rate of the last few years. In addition, a new children's menu will be introduced as well as 'Max Havelaar' fair-trade coffee at a price of 1.70 CHF.

  12. Alteraciones histológicas en riñones de ratas, inducidas por dosis bajas del veneno de la serpiente Bothrops asper. - Histological injuries in rat’s kidneys induced to low dose of snake’s poison Bothrops asper.

    Directory of Open Access Journals (Sweden)

    Reyes, Adriana

    2009-09-01

    Full Text Available ResumenEl objetivo del trabajo fue demostrar que dosis bajas del veneno deBothrops asper pueden causar patologías renales que se acentúancon el tiempo de exposición al veneno, especialmente a nivelglomérular. Se emplearon cinco grupos de ratas de cinco individuoscada uno, los cuales fueron inoculados con una dosis de veneno de2.1µg/g de peso (por vía intraperitoneal, con adición de un grupocontrol, al cual se le inyectó solución salina. A los animales se lespracticó la eutanasia secuencialmente (1, 48, 96, 144 y 288 horas.En cada tiempo se extrajeron los riñones del grupo seleccionado, paraprocesarlos con técnicas de coloración (HE – PAS. Con el diagnósticohistopatológico se evidenció en la primera hora gloméruloscongestionados con tumefacción, después de 48 horas se presentóuna glomérulonefritis, seguida de un colapso glomerular a las 96horas. Las patologías se evidenciaron mas en los últimos períodos deexperimentación; a las 144 horas se observó glomérulonefritismembranosa y proliferativa endocapilar culminando con la desaparición de la cápsula de Bowman y la estructura total del glomérulo a las 288 horas. Se concluye que cantidades bajas del veneno pueden causar patologías renales progresivas con el paso del tiempo.SummaryAim of this work was to demonstrate that low doses of Bothropsasper venom can cause pathologies that progress over time on therenal tissue, at the glomerular level especially. Five groups of ratsconsisting of five individuals were used and a single dose of poison2.1µg/g in weight was injected. Control group was injected withsaline solution. Euthanasia sequentially was practiced (1, 48, 96, 144and 288 hours. In each time was extracted the kidneys of groupselection, to procession with color’s techniques (HE – PAS.Histopathological diagnostic evidenced congested glomeruli withswelling in the first hour, after 48 hours a glomerulonephritis waspresented, followed by a collapsed glomerular at

  13. Purification and Characterization of F Ⅱ A, a Novel Prothrombin-activator from Bothrops atrox Venom%一种新的蛇毒凝血酶原激活物的分离纯化及特征

    Institute of Scientific and Technical Information of China (English)

    孙东; 石皎; 丁忠福; 李秀琳; 李秀娜; 崔亮亮; 薛雁

    2011-01-01

    In this study, a novel prothrombin-activator from the venom of Bothrops atrox designated F II A was isolated. F H A was purified using a SP Sepharose Fast Flow anion-exchange chromatography, DEAE-Sepharose Fast Flow cation-exchange chromatography followed by SP Sepharose High Performance anion-exchange chromatography. F II A was a single-chain polypeptide with a molecular mass 72 kD as measured by reduced SDS-PAGE. Its pi was 6. 67 as measured by IEF. Its N-terminal amino acid sequence was ALVLIAFAQVLQQCP and the accession number was B3A0N1 in UniProtKB Database. The enzymatic activity of F II A was inhibited by the chelating EDTA, whereas the serine protease inhibitor PMSF had no effect on its activity. Interestingly, its enzymatic activity is not requiring cofactors such as Ca2+ , Fva or phospholipid. The results suggest that F II A is a P- I metalloproteinase and its pathways for prothrombin activation is similar to F X a.%为获得矛头蝮蛇(Bothrops atrox)蛇毒凝血酶原激活物并研究其基本性质,采用SP Sepharose Fast Flow,DEAE-Sepharose Fast Flow和SP Sepharose High Performance等层析方法从巴西矛头蝮蛇蛇毒中分离纯化得到1种单一组分的凝血酶原激活物(prothrombin activator,FⅡA).还原性SDSPAGE结果显示,其分子质量约为72 kD,等电点为6.67.HPSEC显示纯度大于95%.该酶是1种N连接的糖蛋白,N末端氨基酸序列为ALVLIAFAQYLQQCP,获得登录号为:B3A0N1.其活性可被EDTA-Na2抑制,PMSF对其活性无影响,对凝血酶原的激活过程无需Ca2+、FVa、磷脂的参与,为P-I金属蛋白酶,对凝血酶原的激活方式与FXa相似.本研究纯化与鉴定的新凝血酶原激活物为其药学研究及临床应用提供参考.

  14. Fluorometric assay using naphthylamide substrates for assessing novel venom peptidase activities.

    Science.gov (United States)

    Gasparello-Clemente, Elaine; Silveira, Paulo Flávio

    2002-11-01

    In the present study we examined the feasibility of using the fluorometry of naphthylamine derivatives for revealing peptidase activities in venoms of the snakes Bothrops jararaca, Bothrops alternatus, Bothrops atrox, Bothrops moojeni, Bothrops insularis, Crotalus durissus terrificus and Bitis arietans, of the scorpions Tityus serrulatus and Tityus bahiensis, and of the spiders Phoneutria nigriventer and Loxosceles intermedia. Neutral aminopeptidase (APN) and prolyl-dipeptidyl aminopeptidase IV (DPP IV) activities were presented in all snake venoms, with the highest levels in B. alternatus. Although all examined peptidase activities showed relatively low levels in arthropod venoms, basic aminopeptidase (APB) activity from P. nigriventer venom was the exception. Compared to the other peptidase activities, relatively high levels of acid aminopeptidase (APA) activity were restricted to B. arietans venom. B. arietans also exhibited a prominent content of APB activity which was lower in other venoms. Relatively low prolyl endopeptidase and proline iminopeptidase activities were, respectively, detectable only in T. bahiensis and B. insularis. Pyroglutamate aminopeptidase activity was undetectable in all venoms. All examined peptidase activities were undetectable in T. serrulatus venom. In this study, the specificities of a diverse array of peptidase activities from representative venoms were demonstrated for the first time, with a description of their distribution which may contribute to guiding further investigations. The expressive difference between snake and arthropod venoms was indicated by APN and DPP IV activities while APA and APB activities distinguished the venom of B. arietans from those of Brazilian snakes. The data reflected the relatively uniform qualitative distribution of the peptidase activities investigated, together with their unequal quantitative distribution, indicating the evolutionary divergence in the processing of peptides in these different

  15. Purificación y caracterización de la l-amino ácido oxidasa del veneno de la serpiente Bothrops brazili "jergón shushupe"

    Directory of Open Access Journals (Sweden)

    Christian Solís

    2014-06-01

    Full Text Available Se ha purificado y caracterizado parcialmente la L-aminoácido oxidasa de la serpiente Bothrops brazili. El aislamiento se realizó usando técnicas cromatográficas en Sephadex G-100 y CM-Sephadex C-50, utilizando como eluyente buffer acetato de amonio 0,1M pH 6. La enzima fue purificada 29,3 veces con un rendimiento de 30,9%. Usando electroforesis en gel de poliacrilamida con dodecil sulfato de sodio (PAGE-SDS en condiciones reductoras y no reductoras, así como las técnicas de inmunodifusión e inmunoelectroforesis, se demostró la presencia de una sola banda proteica. La enzima fue caracterizada como una glicoproteína ácida con un peso molecular de 125,7 kd formada por dos subunidades de 59,9 kd unidas por enlaces débiles, con un pH óptimo entre 7,5 y 9, dependiendo del aminoácido usado como substrato; siendo termoestable hasta los 550 C y lábil a pH alcalino. Asimismo, los ensayos por el método del cilindro en placa de Grove demostraron el efecto antibacteriano de la proteína aislada en cepas estandarizadas de Staphylococcus aureus, Vibrio cholerae y Streptococcus faecalis.

  16. Interaction between TNF and BmooMP-Alpha-I, a Zinc Metalloprotease Derived from Bothrops moojeni Snake Venom, Promotes Direct Proteolysis of This Cytokine: Molecular Modeling and Docking at a Glance

    Science.gov (United States)

    Silva, Maraisa Cristina; Lopes Silva, Tamires; Silva, Murilo Vieira; Mota, Caroline Martins; Santiago, Fernanda Maria; Fonseca, Kelly Cortes; Oliveira, Fábio; Mineo, Tiago Wilson Patriarca; Mineo, José Roberto

    2016-01-01

    Tumor necrosis factor (TNF) is a major cytokine in inflammatory processes and its deregulation plays a pivotal role in several diseases. Here, we report that a zinc metalloprotease extracted from Bothrops moojeni venom (BmooMP-alpha-I) inhibits TNF directly by promoting its degradation. This inhibition was demonstrated by both in vitro and in vivo assays, using known TLR ligands. These findings are supported by molecular docking results, which reveal interaction between BmooMP-alpha-I and TNF. The major cluster of interaction between BmooMP-alpha-I and TNF was confirmed by the structural alignment presenting Ligand Root Mean Square Deviation LRMS = 1.05 Å and Interactive Root Mean Square Deviation IRMS = 1.01 Å, this result being compatible with an accurate complex. Additionally, we demonstrated that the effect of this metalloprotease on TNF is independent of cell cytotoxicity and it does not affect other TLR-triggered cytokines, such as IL-12. Together, these results indicate that this zinc metalloprotease is a potential tool to be further investigated for the treatment of inflammatory disorders involving TNF deregulation. PMID:27447669

  17. Two phospholipase A2 inhibitors from the plasma of Cerrophidion (Bothrops) godmani which selectively inhibit two different group-II phospholipase A2 myotoxins from its own venom: isolation, molecular cloning and biological properties.

    Science.gov (United States)

    Lizano, S; Angulo, Y; Lomonte, B; Fox, J W; Lambeau, G; Lazdunski, M; Gutiérrez, J M

    2000-01-01

    Myotoxic phospholipases A(2) (PLA(2)s; group II) account for most of the muscle-tissue damage that results from envenomation by viperid snakes. In the venom of the Godman's viper (Cerrophidion godmani, formerly Bothrops godmani), an enzymically active PLA(2) (myotoxin I) and an inactive, Lys-49 variant (myotoxin II) induce extensive muscle damage and oedema. In this study, two distinct myotoxin inhibitor proteins of C. godmani, CgMIP-I and CgMIP-II, were purified directly from blood plasma by selective binding to affinity columns containing either myotoxin I or myotoxin II, respectively. Both proteins are glycosylated, acidic (pI=4) and composed of 20-25-kDa subunits that form oligomers of 110 kDa (CgMIP-I) or 180 kDa (CgMIP-II). In inhibition studies, CgMIP-I specifically neutralized the PLA(2) and the myotoxic, oedema-forming and cytolytic activities of myotoxins I, whereas CgMIP-II selectively inhibited the toxic properties of myotoxin II. N-terminal amino acid sequence analysis and sequencing of cDNAs encoding the two inhibitors revealed that CgMIP-I is similar to gamma-type inhibitors, which share a pattern of cysteine residues present in the Ly-6 superfamily of proteins, whereas CgMIP-II shares sequence identity with alpha-type inhibitors that contain carbohydrate-recognition-like domains, also found in C-type lectins and mammalian PLA(2) receptors. N-terminal sequencing of myotoxin I revealed a different primary structure from myotoxin II [De Sousa, Morhy, Arni, Ward, Díaz and Gutiérrez (1998) Biochim. Biophys. Acta 1384, 204-208], which provides insight into the nature of such pharmacological specificity. PMID:10698689

  18. Glycolic acid inhibits enzymatic, hemorrhagic and edema-inducing activities of BaP1, a P-I metalloproteinase from Bothrops asper snake venom: insights from docking and molecular modeling.

    Science.gov (United States)

    Pereañez, Jaime Andrés; Patiño, Arley Camilo; Rey-Suarez, Paola; Núñez, Vitelbina; Henao Castañeda, Isabel Cristina; Rucavado, Alexandra

    2013-09-01

    Glycolic acid (GA) (2-Hydroxyethanoic acid) is widely used as chemical peeling agent in Dermatology and, more recently, as a therapeutic and cosmetic compound in the field of skin care and disease treatment. In this work we tested the inhibitory ability of glycolic acid on the enzymatic, hemorrhagic and edema-inducing activities of BaP1, a P-I metalloproteinase from Bothrops asper venom, which induces a variety of toxic actions. Glycolic acid inhibited the proteolytic activity of BaP1 on azocasein, with an IC₅₀ of 1.67 mM. The compound was also effective at inhibiting the hemorrhagic activity of BaP1 in skin and muscle in experiments involving preincubation of enzyme and inhibitor prior to injection. When BaP1 was injected i.m. and then, at the same site, different concentrations of glycolic acid were administered at either 0 or 5 min, 7 mM solutions of the inhibitor partially abrogated hemorrhagic activity when administered at 0 min. Moreover, glycolic acid inhibited, in a concentration-dependent manner, edema-forming activity of BaP1 in the footpad. In order to have insights on the mode of action of glycolic acid, UV-vis and intrinsic fluorescence studies were performed. Results of these assays suggest that glycolic acid interacts directly with BaP1 and chelates the Zn²⁺ ion at the active site. These findings were supported by molecular docking results, which suggested that glycolic acid forms hydrogen bonds with residues Glu143, Arg110 and Ala111 of the enzyme. Additionally, molecular modeling results suggest that the inhibitor chelates Zn²⁺, with a distance of 3.58 Å, and may occupy part of substrate binding cleft of BaP1. Our results suggest that glycolic acid is a candidate for the development of inhibitors to be used in snakebite envenomation.

  19. Variaciones ontogénicas y geográficas en los efectos tóxicos y en las características inmunoquímicas del veneno de serpientes del grupo Bothrops atrox del Meta y Antioquia

    Directory of Open Access Journals (Sweden)

    María Fabiola Toro

    2000-02-01

    Full Text Available

    El accidente ofídico es un problema de salud pública frecuente en algunos lugares del mundo. En Latinoamérica, el género Bothrops es el causante del 90% de los accidentes ofídicos y en Colombia el grupo Bothrops atrox es responsable del 50% o más de los casos que se reportan anualmente (2000 casos. Este grupo se encuentra distribuido en la mayor parte del territorio nacional colombiano, incluyendo el este y oeste de los Andes, vertiente Pacífica y Atlántica, además de los valles interandinos.

    El veneno del grupo Bothrops atrox induce un complejo cuadro de alteraciones fisiopatológicas locales y sistémicas, como mionecrosis, dermonecrosis, edema, hemorragia, dolor, coagulopatías, choque cardiovascular, nefrotoxicidad, actividad proteolítica y hemolítica indirecta.

    En diversos países se han realizado estudios sobre la variabilidad de los venenos de diferentes especies de serpientes. Se han encontrado diferencias en todos los niveles taxonómicos, siendo de mayor importancia la variabilidad intraespecífica. Con este estudio se pretende determinar la variabilidad ontogénica y geográfica del veneno de Bothrops atrox procedente de dos regiones colombianas, mediante pruebas biológicas e inmunoquímicas. Los resultados previos muestran que existe una variación ontogénica en el veneno de la especie Bothrops atrox del departamento del Meta con respecto a la letalidad, y a las actividades hemorrágica, coagulante y edematizante. Las serpientes recién nacidas tienen una mayor letalidad (DL50 = 51.8 ± 5.6 mg de veneno con respecto a los adultos (81.4±

  20. Características bioquímicas y capacidad neutralizante de cuatro antivenenos polivalentes frente a los efectos farmacológicos y enzimáticos del veneno de Bothrops asper y Porthidium nasutum de Antioquia y Chocó

    Directory of Open Access Journals (Sweden)

    Mónica Saldarriaga

    2002-01-01

    Full Text Available En Colombia, el 90-95% de las 3000 mordeduras de serpientes informadas cada año, son ocasionadas por Bothrops spp, con una elevada mortalidad y secuelas. Siguiendo recomendaciones de la OMS, se evaluó la capacidad neutralizante de los efectos farmacológicos y enzimáticos de los venenos de Bothrops asper y Porthidium nasutum de Antioquia y Chocó por cuatro antivenenos; 2 de ellos de IgG completa (polivalente antibothrópico, anticrotálico del Instituto Nacional de Salud INS -Colombia; polivalente antibothrópico, anticrotálico, antilachésico de Laboratorios Probiol -Colombia y 2 antivenenos de fragmentos F(ab’2 (polivalente antibothrópico, anticrotálico del Centro de Biotecnología de la Universidad Central de Venezuela; y el polivalente antibothrópico, anticrotálico Antivipmyn® del Instituto Bioclón -México. Se determinó la actividad letal, hemorrágica, desfibrinante, edematizante, mionecrosante y hemolítica indirecta de cada veneno, siguiendo métodos ya estandarizados. Las pruebas de neutralización in vitro e in vivo se realizaron por el método de preincubación a 370C de dosis fijas de veneno y dosis variables de antiveneno. Los antivenenos Antivipmyn® de México y polivalente INS de Colombia tuvieron la mayor potencia neutralizante de todos los efectos farmacológicos y enzimáticos del veneno de B. asper y P. nasutum. El antiveneno polivalente Probiol fue el de menor capacidad neutralizante y mayor concentración de proteínas. Los antivenenos de fragmentos F(ab’2 tuvieron más baja concentración de proteínas y solo cantidades menores de proteínas no inmunes por electroforesis. Ninety to 95% of the snakebites reported yearly in Colombia are inflicted by Bothrops spp with high mortality and sequelae. Following recommendations of the World Health Organization, the neutralizing ability of four polyvalent antivenoms against several pharmacological and enzymatic effects of Bothrops asper and Porthidium nasutum snake

  1. Bothrops bites in Colombia: a multicenter study on the efficacy and safety of Antivipmyn-Tri®, a polyvalent antivenom produced in Mexico Accidente bothrópico en Colombia: estudio multicéntrico de la eficacia y seguridad de Antivipmyn-Tri®, un antiveneno polivalente producido en México

    Directory of Open Access Journals (Sweden)

    María Eugenia Espinal Saldarriaga

    2007-08-01

    Full Text Available Introduction: Colombia is a country with two whole IgG antivenom producers, but the expectancy of all the market are not fulfilled by different technical reasons. Objectives: To evaluate the efficacy and safety of a F(ab´2 polyvalent antivenom produced in Mexico and of a new dosage regimen for Bothrops bites in Colombia. Methods: A clinical trial, including serum venom and antivenom measurements (ELISA, was performed during 9 months in 53 patients. Results: forty four patients were bitten by Bothrops asper in Antioquia and Chocó and 9 by B. atrox in Amazonas; on admission, all of them had nonclottable blood, 30 (56.6% presented local and 24 (45.3% systemic bleeding. The final envenoming grade was mild in 13 (24.5%, moderate in 30 (56.6% and severe in 10 patients (18.9%. At the antivenom doses used in this study (5 vials for mild / moderate and 10 for severe envenoming, Antivipmyn Triwas 100% efficient to decrease significantly serum venom concentrations within the first treatment hour, and to stop local and systemic bleeding within 6-12 hours, 96.2% efficient to restore blood coagulation within 24 hours and 100% within 48 hours. Two patients (3.8% had recurrence of coagulopathy without bleeding, and there were 12 recurrences of antigenaemia without clinical relevance. Ten (18.9% patients suffered early mild adverse reactions to fabotherapy. There were no deaths and four patients (7.5% presented sequelae. Conclusion: at the doses used in this study, Antivipmyn Tri® was efficient and safe for the treatment of Bothrops bites in Colombia. Introducción: Colombia es un país con dos productores de antivenenos de IgG, pero hay un mercado insatisfecho por diferentes razones técnicas. Objetivos: evaluar la eficacia y la seguridad de un antiveneno F(ab´2 polivalente (Antivipmyn-Tri® producido en México, y un nuevo esquema de dosis en accidente bothrópico en Colombia. Métodos: se realizaron durante 9 meses un ensayo clínico-terapéutico y

  2. VARIACIONES EN L COMPOSICIÓN PROTEICA, ACTIVIDADES ENZIMÁTICAS Y BIOLÓGICAS DEL VENENO DE LA SERPIENTE BOTHROPS ATROX (VIPEREIDAE EN RELACIÓN CON LA EDAD

    Directory of Open Access Journals (Sweden)

    O. Málaga

    2014-06-01

    Full Text Available La serpiente Bothrops atrox, que habita en la Selva Amazónica del Perú, es responsable del mayor número de accidentes ofídicos y constituye un problema de salud pública. Por esta razón, se ha realizado un estudio de las variaciones en la composición y actividad del veneno en tres grupos de especimenes en cautiverio, que corresponden a juveniles de 1 y 2 años, así como ejemplares adultos mayores de 5 años. Los venenos obtenidos fueron liofilizados para su conservación y diluidos en concentraciones iniciales de 1 mg/ml en solución salina o en el buffer apropiado. Con estas muestras se efectuaron los análisis de concentración de proteína por D. 0. 280 nm y por el método de Lowry, determinándose además el número de bandas proteicas por electroforesis en gel de poliacrilamida con dodecil sulfato de sodio (PAGE-SIDS. Con las mismas muestras se hicieron ensayos para medir las siguientes actividades enzimáticas: caseinolítica, amidolftica, coagulante, hialuronidasa, L-aminocácido oxidasa (LAO y fosfolipasa A, adicionalmente se determinó la actividad hemorrágica y el efecto edemático con ratones albinos. Los resultados mostraron que la mayor concentración proteica (0,938mg de proteína/mg de veneno así como el número máximo de bandas proteicas (8 bandas se obtuvieron con los venenos de juveniles de 2 años. Asimismo se encontró que algunas actividades enzimáticas como la amidolítica, fosfolipásica y L-aminoácido oxidasa fueron más altas en los juveniles de 2 años, además de los efectos hemorrágico y edemático. En cambio, las actividades coagulante y proteolítica sobre caseína se elevaron progresivamente con la edad de los ejemplares. Sin embargo, la enzima hialuronidasa tuvo su máximo valor en los juveniles de 1 año, decreciendo notablemente en los adultos. Es interesante señalar también que fosfolipasa A2 en los adultos sólo tuvo una actividad de 6,4% con respecto al valor más alto encontrado en el veneno

  3. A comparison of the ability of Bellucia dichotoma Cogn. (Melastomataceae) extract to inhibit the local effects of Bothrops atrox venom when pre-incubated and when used according to traditional methods.

    Science.gov (United States)

    Mourão de Moura, Valéria; Serra Bezerra, Adrielle N; Veras Mourão, Rosa Helena; Varjão Lameiras, Juliana L; Almeida Raposo, Juliana D; Luckwu de Sousa, Rafael; Boechat, Antônio Luiz; Bezerra de Oliveira, Ricardo; de Menezes Chalkidis, Hipocrátes; Dos-Santos, Maria Cristina

    2014-07-01

    Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd

  4. A comparison of the ability of Bellucia dichotoma Cogn. (Melastomataceae) extract to inhibit the local effects of Bothrops atrox venom when pre-incubated and when used according to traditional methods.

    Science.gov (United States)

    Mourão de Moura, Valéria; Serra Bezerra, Adrielle N; Veras Mourão, Rosa Helena; Varjão Lameiras, Juliana L; Almeida Raposo, Juliana D; Luckwu de Sousa, Rafael; Boechat, Antônio Luiz; Bezerra de Oliveira, Ricardo; de Menezes Chalkidis, Hipocrátes; Dos-Santos, Maria Cristina

    2014-07-01

    Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd

  5. Presynaptic Proteins as Markers of the Neurotoxic Activity of BmjeTX-I and BmjeTX-II Toxins from Bothrops marajoensis (Marajó Lancehead) Snake Venom

    Science.gov (United States)

    Lisboa, Antonio; Melaré, Rodolfo; Franco, Junia R. B.; Bis, Carolina V.; Gracia, Marta; Ponce-Soto, Luis A.; Marangoni, Sérgio; Rodrigues-Simioni, Léa; da Cruz-Höfling, Maria Alice

    2016-01-01

    Neuromuscular preparations exposed to B. marajoensis venom show increases in the frequency of miniature end-plate potentials and twitch tension facilitation followed by presynaptic neuromuscular paralysis, without evidences of muscle damage. Considering that presynaptic toxins interfere into the machinery involved in neurotransmitter release (synaptophysin, synaptobrevin, and SNAP25 proteins), the main objective of this communication is to analyze, by immunofluorescence and western blotting, the expression of the synaptic proteins, synaptophysin, synaptobrevin, and SNAP25 and by myography, light, and transmission electron microscopy the pathology of motor nerve terminals and skeletal muscle fibres of chick biventer cervicis preparations (CBC) exposed in vitro to BmjeTX-I and BmjeTX-II toxins from B. marajoensis venom. CBC incubated with toxins showed irreversible twitch tension blockade and unaffected KCl- and ACh-evoked contractures, and the positive colabelling of acetylcholine receptors confirmed that their action was primarily at the motor nerve terminal. Hypercontraction and loose myofilaments and synaptic vesicle depletion and motor nerve damage indicated that the toxins displayed both myotoxic and neurotoxic effect. The blockade resulted from interference on synaptophysin, synaptobrevin, and SNAP25 proteins leading to the conclusion that BmjeTX-I and BmjeTX-II affected neurotransmitter release machinery by preventing the docking of synaptic vesicles to the axolemma of the nerve terminal. PMID:27635261

  6. Increases in Problem Drinking

    Science.gov (United States)

    ... Increases in Problem Drinking Join the Fight Against Superbugs Featured Website: Genetic and Rare Diseases Information Center ... Increases in Problem Drinking Join the Fight Against Superbugs Featured Website: Genetic and Rare Diseases Information Center ...

  7. Effect of increased

    NARCIS (Netherlands)

    Carreira , C.; Heldal, M.; Bratbak, G.

    2013-01-01

    Atmospheric carbon dioxide (CO2) has increased since the pre-industrial period and is predicted to continue to increase throughout the twenty-first century. The ocean is a sink for atmospheric CO2 and increased CO2 concentration will change the carbonate equilibrium of seawater and result in lower c

  8. Increasing SLEDed Linac Gradient

    Energy Technology Data Exchange (ETDEWEB)

    Farkas, Zoltan D

    2001-11-08

    This note will show how to increase the SLED [1] gradient by varying Q{sub e}, the external Q of the SLED cavity, by increasing its Q{sub 0} and by increasing the compression ratio. If varying the external Q is to be effective, then the copper losses should be small so that Q{sub 0} >> Q{sub e}. Methods of varying Q{sub e} will be indicated but no experimental data will be presented. If we increase the klystron pulse width from 3.5 to 5 {micro}S and increase Q{sub 0} from the present 100000 to 300000, then the gradient increases by 19% and the beam energy increases from 50 to 60 GeV. This note will also discuss SLED operation at 11424 MHz, the NLC frequency. Without Q{sub e} switching, using SLED at 11424 MHz increases the SLAC gradient from 21 MV/m to 34 MV/m, and at the same repetition rate, uses about 1/5 of rf average power. If we also double the compression ratio, we reach 47 MV/m and over 100 GeV beam energy.

  9. Increasing productivity: Another approach

    Energy Technology Data Exchange (ETDEWEB)

    Norton, F.J.

    1996-06-10

    An engineering information (EI) and information technology (IT) organization that must improve its productivity should work to further its business goals. This paper explores a comprehensive model for increasing EI/IT productivity by supporting organizational objectives.

  10. Increasing hygiene productivity.

    Science.gov (United States)

    Levin, Roger P

    2003-03-01

    Dentists have many opportunities to expand the role of dental hygienists and provide patients with better oral health care, while increasing production and profits. But the proper business systems and verbal skills need to be incorporated. You must train hygienists to do all they can do for every patient. Begin with one service and add others, as the hygienists becomes familiar with each one. Set a goal of a 15% increase in production per year for the hygiene department. Clinicians using these strategies have experienced as much as a 100% to 200% increase in hygiene revenue during the first year of incorporating these services. An added benefit is that these dentists often see a substantial increase in dental treatment diagnosis and case acceptance. An effective and efficient hygiene department will often identify and help secure more than 50% of a doctor's production.

  11. Increasing Public Library Productivity.

    Science.gov (United States)

    Samuelson, Howard

    1981-01-01

    Suggests ways of improving productivity for public libraries faced with increased accountability, dwindling revenues, and continuing inflation. Techniques described include work simplification, work analysis, improved management, and employee motivation. (RAA)

  12. Exploding Increase of Population

    Energy Technology Data Exchange (ETDEWEB)

    Kim, J.H. [Sunmoon University, Chonan (Korea)

    2002-03-01

    Until 1650, the population of the world did not increase properly. According to studies of the demography, the annual increase rate of the world population during 2500 years, from 850 B.C. to 1650 A.D., was just 0.07%. Currently, however, the world population, which has exceptionally rapidly increased from 1900, is more than 6 billion as of 2000. After World War II, especially, the increase rate of the population has risen to about 1.8%, so we can use the word, explosion of the population. The explosion of the population accompanies the increase of energy consumption. The energy production of every year cannot sufficiently meet the energy demand, so we can face the grand energy crisis someday. The date might be a someday after 2020. According to the future forecasting of Shell, one of the majors, the peak of oil supply will be between 2015 and 2020. Unless the alternative energy is developed, the whole world will suffer the serious oil crisis.

  13. Seismic waves increase permeability.

    Science.gov (United States)

    Elkhoury, Jean E; Brodsky, Emily E; Agnew, Duncan C

    2006-06-29

    Earthquakes have been observed to affect hydrological systems in a variety of ways--water well levels can change dramatically, streams can become fuller and spring discharges can increase at the time of earthquakes. Distant earthquakes may even increase the permeability in faults. Most of these hydrological observations can be explained by some form of permeability increase. Here we use the response of water well levels to solid Earth tides to measure permeability over a 20-year period. At the time of each of seven earthquakes in Southern California, we observe transient changes of up to 24 degrees in the phase of the water level response to the dilatational volumetric strain of the semidiurnal tidal components of wells at the Piñon Flat Observatory in Southern California. After the earthquakes, the phase gradually returns to the background value at a rate of less than 0.1 degrees per day. We use a model of axisymmetric flow driven by an imposed head oscillation through a single, laterally extensive, confined, homogeneous and isotropic aquifer to relate the phase response to aquifer properties. We interpret the changes in phase response as due to changes in permeability. At the time of the earthquakes, the permeability at the site increases by a factor as high as three. The permeability increase depends roughly linearly on the amplitude of seismic-wave peak ground velocity in the range of 0.21-2.1 cm s(-1). Such permeability increases are of interest to hydrologists and oil reservoir engineers as they affect fluid flow and might determine long-term evolution of hydrological and oil-bearing systems. They may also be interesting to seismologists, as the resulting pore pressure changes can affect earthquakes by changing normal stresses on faults. PMID:16810253

  14. Does salt increase thirst?

    Science.gov (United States)

    Leshem, Micah

    2015-02-01

    Our diet is believed to be overly rich in sodium, and it is commonly believed that sodium intake increases drinking. Hence the concern of a possible contribution of dietary sodium to beverage intake which in turn may contribute to obesity and ill health. Here we examine whether voluntary, acute intake of a sodium load, as occurs in routine eating and snacking, increases thirst and drinking. We find that after ingesting 3.5 or 4.4 g NaCl (men) and 1.9 or 3.7 g (women) on nuts during 15 minutes, there is no increase in thirst or drinking of freely available water in the following 2 h compared with eating similar amounts of sugared or unflavored nuts. This suggests that routine ingestion of boluses of salt (~30-40% of daily intake for men, ~ 20-40% for women) does not increase drinking. Methodological concerns such as about nuts as vehicle for sodium suggest further research to establish the generalizability of this unexpected result. PMID:25447020

  15. Increased urinary orosomucoid excretion

    DEFF Research Database (Denmark)

    Christiansen, M S; Iversen, K; Larsen, C T;

    2009-01-01

    OBJECTIVE: In a previous study, urinary orosomucoid excretion rate (UOER) independently predicted cardiovascular mortality in patients with type 2 diabetes. The aim of the present study was to determine whether increased UOER is associated with cardiovascular risk factors such as inflammation, im...

  16. Increasing income inequality

    DEFF Research Database (Denmark)

    Frederiksen, Anders; Poulsen, Odile

    In recent decades most developed countries have experienced an increase in income inequality. In this paper, we use an equilibrium search framework to shed additional light on what is causing an income distribution to change. The major benefit of the model is that it can accommodate shocks...... to the skill composition in the market, employee bargaining power and productivity. Further, when our model is subjected to skill-upgrading and changes in employee bargaining power, it is capable of predicting the recent changes observed in the Danish income distribution. In addition, the model emphasizes...... that shocks to the employees' relative productivity, i.e., skill-biased technological change, are unlikely to have caused the increase in income inequality....

  17. Pemex increasing offshore activity

    Energy Technology Data Exchange (ETDEWEB)

    Beachy, D.

    1985-05-01

    Although austere by boom-year standards, Mexico's National Energy Program for 1984-1988 calls for forty wildcats and 90 to 144 development wells off the coast, primarily in the prolific Campeche Bay area. Platform additions will include nine drilling platforms, each for twelve wells, and eight eight platforms to drill injection wells. Additionally, 7 production, 6 accomodation, 6 linkage and 8 compression platforms and 13 tetrapods will be installed. The main objectives of the plan are energy self-sufficiency through the turn of the century, and energy diversification, savings and productivity. The most controversial portion of Mexico's energy program is that calling for nuclear energy development. The energy program lists three basic goals in hydrocarbon production: continuing research on better techniques of secondary recovery; increasing capacity for refining primary and secondary crude products and improving production of heavy crudes; and increasing storage capacity and installing pipelines capable of carrying a greater volume of crude.

  18. High population increase rates.

    Science.gov (United States)

    1991-09-01

    In addition to its economic and ethnic difficulties, the USSR faces several pressing demographic problems, including high population increase rates in several of its constituent republics. It has now become clear that although the country's rigid centralized planning succeeded in covering the basic needs of people, it did not lead to welfare growth. Since the 1970s, the Soviet economy has remained sluggish, which as led to increase in the death and birth rates. Furthermore, the ideology that held that demography could be entirely controlled by the country's political and economic system is contradicted by current Soviet reality, which shows that religion and ethnicity also play a significant role in demographic dynamics. Currently, Soviet republics fall under 2 categories--areas with high or low natural population increase rates. Republics with low rates consist of Christian populations (Armenia, Moldavia, Georgia, Byelorussia, Russia, Lithuania, Estonia, Latvia, Ukraine), while republics with high rates are Muslim (Tadzhikistan, Uzbekistan, Turkmenistan, Kirgizia, Azerbaijan Kazakhstan). The later group has natural increase rates as high as 3.3%. Although the USSR as a whole is not considered a developing country, the later group of republics fit the description of the UNFPA's priority list. Another serious demographic issue facing the USSR is its extremely high rate of abortion. This is especially true in the republics of low birth rates, where up to 60% of all pregnancies are terminated by induced abortions. Up to 1/5 of the USSR's annual health care budget is spent on clinical abortions -- money which could be better spent on the production of contraceptives. Along with the recent political and economic changes, the USSR is now eager to deal with its demographic problems. PMID:12284289

  19. Increase in family allowances.

    Science.gov (United States)

    1989-01-01

    In July 1989 the family allowance structure in Australia was changed from a 4-rate to a 2-rate structure. The new rates were increased to $A9 a week for the 1st 3 children and $A12 for each additional child. The Family Allowance Supplment rate for children 13-15 years old was raised from $A31 to $A34.10/week. PMID:12344544

  20. Market Expects Demand Increase

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    In the recent releasing Textile Industry Invigorating Plan,"givingattention to both domestlc and overseas markets"is put into a keyposition.Under a series policies,such as increasing the tax rebaterate for textile and garment exports,and granting loan for SME,thefurther development of this industry is expectative.Otherwise,weshould know that it costs time for demand driving.This need ourpatients.The only questionis how much time we have to wait.

  1. Increasing student success

    Science.gov (United States)

    Stewart, Gay; Stewart, John

    2013-03-01

    A more scientifically literate society benefits all STEM disciplines, as well as society as a whole. It is best realized by better serving all undergraduate STEM students. In better-serving all students, a physics department also benefits. The University of Arkansas, Fayetteville physics department has seen a drastic change in number of majors, the number of students active in research and the number of graduates pursuing graduate work, while also increasing the number of majors who decide to teach. Prior to our involvement with the Physics Teacher Education Coalition, graduation rates had increased by more than a factor of 4 in 4 years. After the increased efforts when we became a part of PhysTEC (http://PhysTEC.org) our graduation numbers doubled again. Specific attention to class policy to impact student learning in our introductory courses and strong preparation of the graduate teaching assistants, and quality advising were our primary areas of emphasis. What worked to build these numbers and strengthen these resources at Arkansas will be discussed. This work was supported in part by the National Science Foundation and through the Physics Teacher Education Coalition.

  2. Condom use increasing.

    Science.gov (United States)

    Finger, W R

    1998-01-01

    Condom use is central to the prevention of AIDS among people at risk for contracting HIV. As such, condom use is increasing dramatically even though many men say that they do not like using them. Condom sales through social marketing campaigns have increased dramatically in some countries, where tens of millions of condoms are sold annually. For example, during the period 1991-96, annual social marketing sales increased about five-fold in Ethiopia to 21 million, and nine-fold in Brazil to 27 million. These sales reflect the success of condom social marketing campaigns in making condoms accessible and largely affordable. There is also a greater general awareness of AIDS than there used to be, and communication campaigns have shown that condoms are an effective solution. More condoms still need to be used in the ongoing struggle against HIV/AIDS. The author discusses the factors which affect the limited acceptance of condoms, condom use outside of marriage, social marketing, and family planning programs. PMID:12293530

  3. Alarming increase in refugees.

    Science.gov (United States)

    1992-01-01

    Over the past decade and half there has been an alarming worldwide increase in refugees. The total rose form 2.8 million in 1976 to 8.2 million in 1980, to 17.3 million in 1990. Africa's refugees rose from 1.2 million in 1976 to 5.6 million in 1990. Asia's increase over this period was much more rapid--from a mere 180,000 to 8 million. In the Americas the numbers more than trebled, from 770,000 to 2.7 million. Europe was the smallest increase, from 570,000 to 894,000. International law defines a refugee as someone outside of their own country, who has a well-founded fear of persecution because of their political or religious beliefs or ethnic origin, and who cannot turn to their own country for protection. Most refugees are genuine by this definition. The increase reflects, in part, fallout from the cold war. Ethiopia, Mozambique and Angola accounted for almost 1/2 of Africa's refugees; Afghanistan alone for 3/4 of Asia's total. They fled, for the most part, from 1 poor country into another, where they added to shortages of land and fuelwood, and intensified environmental pressure. Malawi, 1 of the poorest countries in the world, is sheltering perhaps as many as 750,000 refugees from the war in Mozambique. But among these refugees--especially among those who turned to the rich countries for asylum--were an increasing number of people who were not suffering political persecution. Driven out of their homes by the collapse of their environment or economic despair, and ready to take any means to get across borders, they are a new category: economic and environmental refugees. The most spectacular attempts hit the television screens: the Vietnamese boat people, ships festooned with Albanians. Behind the headlines there was a growing tide of asylum seekers. The numbers rose 10-fold in Germany from 1983 to 1990. In Switzerland they multiplied by 4 times. In Europe, as a whole, they grew from 71,000 in 1983 to an estimated 550,000 in 1990. In 1990 the numbers threatened to

  4. CHINA INCREASED PETROLEUM TAX

    Institute of Scientific and Technical Information of China (English)

    Mu Xueping

    2005-01-01

    @@ After half a year's gestation and adjustment, the State Administration of Taxation finally set new tax rate of petroleum resources. Notification issued by the Ministry of Finance and the State Administration of Taxation said the petroleum resources tax standard would be lifted nationwide, after the tax rate adjustment, tax on crude oil would be increased to RMB 14-30 yuan per ton and that on natural gas to RMB 7-15 yuan per thousand cubic meters. The new tax rate has been effective nationwide since July 1.

  5. Increasing Childhood Influenza Vaccination

    Science.gov (United States)

    Nowalk, Mary Patricia; Lin, Chyongchiou J.; Hannibal, Kristin; Reis, Evelyn C.; Gallik, Gregory; Moehling, Krissy K.; Huang, Hsin-Hui; Allred, Norma J.; Wolfson, David H.; Zimmerman, Richard K.

    2014-01-01

    Background Since the 2008 inception of universal childhood influenza vaccination, national rates have risen more dramatically among younger children than older children and reported rates across racial/ethnic groups are inconsistent. Interventions may be needed to address age and racial disparities to achieve the recommended childhood influenza vaccination target of 70%. Purpose To evaluate an intervention to increase childhood influenza vaccination across age and racial groups. Methods In 2011–2012, 20 primary care practices treating children were randomly assigned to Intervention and Control arms of a cluster randomized controlled trial to increase childhood influenza vaccination uptake using a toolkit and other strategies including early delivery of donated vaccine, in-service staff meetings, and publicity. Results The average vaccination differences from pre-intervention to the intervention year were significantly larger in the Intervention arm (n=10 practices) than the Control arm (n=10 practices), for children aged 2–8 years (10.2 percentage points (pct pts) Intervention vs 3.6 pct pts Control) and 9–18 years (11.1 pct pts Intervention vs 4.3 pct pts Control, p<0.05), for non-white children (16.7 pct pts Intervention vs 4.6 pct pts Control, p<0.001), and overall (9.9 pct pts Intervention vs 4.2 pct pts Control, p<0.01). In multi-level modeling that accounted for person- and practice-level variables and the interactions among age, race and intervention, the likelihood of vaccination increased with younger age group (6–23 months), white race, commercial insurance, the practice’s pre-intervention vaccination rate, and being in the Intervention arm. Estimates of the interaction terms indicated that the intervention increased the likelihood of vaccination for non-white children in all age groups and white children aged 9–18 years. Conclusions A multi-strategy intervention that includes a practice improvement toolkit can significantly improve influenza

  6. Increasing Inflationary Pressure

    Institute of Scientific and Technical Information of China (English)

    Guo Yan

    2008-01-01

    @@ China's CPI rose to 8.7 percent in February after hitting 7.1 percent in January,creating a 12-year high,the National Bureau of Statistics said.The figure exceeded market expectations,as the Bank of China,the country's secondlargest lender,had predicted a rise of only 8.3 percent.The Consumer Price Index (CPI)is the main gauge of inflation,andthe high CPI over the last two months has put pressure on the government to take action to curb the price increases,and added even more diffculty to the task of reining in this year's inflation rate.

  7. Core Noise - Increasing Importance

    Science.gov (United States)

    Hultgren, Lennart S.

    2011-01-01

    This presentation is a technical summary of and outlook for NASA-internal and NASA-sponsored external research on core (combustor and turbine) noise funded by the Fundamental Aeronautics Program Subsonic Fixed Wing (SFW) Project. Sections of the presentation cover: the SFW system-level noise metrics for the 2015, 2020, and 2025 timeframes; turbofan design trends and their aeroacoustic implications; the emerging importance of core noise and its relevance to the SFW Reduced-Perceived-Noise Technical Challenge; and the current research activities in the core-noise area, with additional details given about the development of a high-fidelity combustor-noise prediction capability as well as activities supporting the development of improved reduced-order, physics-based models for combustor-noise prediction. The need for benchmark data for validation of high-fidelity and modeling work and the value of a potential future diagnostic facility for testing of core-noise-reduction concepts are indicated. The NASA Fundamental Aeronautics Program has the principal objective of overcoming today's national challenges in air transportation. The SFW Reduced-Perceived-Noise Technical Challenge aims to develop concepts and technologies to dramatically reduce the perceived aircraft noise outside of airport boundaries. This reduction of aircraft noise is critical to enabling the anticipated large increase in future air traffic. Noise generated in the jet engine core, by sources such as the compressor, combustor, and turbine, can be a significant contribution to the overall noise signature at low-power conditions, typical of approach flight. At high engine power during takeoff, jet and fan noise have traditionally dominated over core noise. However, current design trends and expected technological advances in engine-cycle design as well as noise-reduction methods are likely to reduce non-core noise even at engine-power points higher than approach. In addition, future low-emission combustor

  8. Increasing Responsibility towards Environment

    Directory of Open Access Journals (Sweden)

    Mihaela Comanescu

    2009-11-01

    Full Text Available In the international climate of the early millennium, the need for reconciliation between economic and social development, on one hand, and the environment, on the other hand, is essential. Therefore, sustainable development must be promoted, because it integrates economic growth, improved quality of life, health, education, social development and environmental protection.Environmental issues are particularly complex as their causes are interdependent. This requires actions at all levels: local, national, regional as well as at the European Union level, each with its own role in increasing responsibility for the environmental protection.It is widely recognized that to obtain successful results an integrated approach has to be used to manage the environment, by adopting long-term strategies and action plans. It requires a detailed analysis of the links between different policies and responsibilities, including the links between different administrative levels.

  9. Increasing Responsibility towards Environment

    Directory of Open Access Journals (Sweden)

    Mihaela COMĂNESCU

    2010-05-01

    Full Text Available In the international climate of the early millennium, the need for reconciliation between economic and social development, on one hand, and the environment, on the other hand, is essential. Therefore, sustainable development must be promoted, because it integrates economic growth, improves quality of life, health, education, social development and environmental protection. Environmental issues are particularly complex as their causes are interdependent. This requires actions at all levels: local, national, regional as well as at the European Union level, each with its own role in increasing responsibility for the environmental protection. It is widely recognized that to obtain successful results an integrated approach has to be used to manage the environment, by adopting long-term strategies and action plans. It requires a detailed analysis of the links between different policies and responsibilities, including the links between different administrative levels.

  10. Elenoside increases intestinal motility

    Institute of Scientific and Technical Information of China (English)

    E Navarro; SJ Alonso; R Navarro; J Trujillo; E Jorge

    2006-01-01

    AIM: To study the effects of elenoside, an arylnaphthalene lignan from Justicia hyssopifolia, on gastrointestinal motility in vivo and in vitro in rats.METHODS: Routine in vivo experimental assessments were catharsis index, water percentage of boluses,intestinal transit, and codeine antagonism. The groups included were vehicle control (propylene glycol-ethanolplant oil-tween 80), elenoside (i.p. 25 and 50 mg/kg),cisapride (i.p. 10 mg/kg), and codeine phosphate (intragastric route, 50 mg/kg). In vitro approaches used isolated rat intestinal tissues (duodenum, jejunum, and ileum). The effects of elenoside at concentrations of 3.2× 10-4, 6.4 × 10-4 and 1.2 × 10-3 mol/L, and cisapride at 10-6 mol/L were investigated.RESULTS: Elenoside in vivo produced an increase in the catharsis index and water percentage of boluses and in the percentage of distance traveled by a suspension of activated charcoal. Codeine phosphate antagonized the effect of 25 mg/kg of elenoside. In vitro, elenoside in duodenum, jejunum and ileum produced an initial decrease in the contraction force followed by an increase.Elenoside resulted in decreased intestinal frequency in duodenum, jejunum, and ileum. The in vitro and in vivo effects of elenoside were similar to those produced by cisapride.CONCLUSION: Elenoside is a lignan with an action similar to that of purgative and prokinetics drugs.Elenoside, could be an alternative to cisapride in treatment of gastrointestinal diseases as well as a preventive therapy for the undesirable gastrointestinal effects produced by opioids used for mild to moderate pain.

  11. Increasing Possibilities of Nanosuspension

    Directory of Open Access Journals (Sweden)

    Kumar Bishwajit Sutradhar

    2013-01-01

    Full Text Available Nowadays, a very large proportion of new drug candidates emerging from drug discovery programmes are water insoluble and thus poorly bioavailable. To avoid this problem, nanotechnology for drug delivery has gained much interest as a way to improve the solubility problems. Nano refers to particles size range of 1–1000 nm. The reduction of drug particles into the submicron range leads to a significant increase in the dissolution rate and therefore enhances bioavailability. Nanosuspensions are part of nanotechnology. This interacts with the body at subcellular (i.e., molecular scales with a high degree of specificity and can be potentially translated into targeted cellular and tissue-specific clinical applications designed to achieve maximal therapeutic efficacy with minimal side effects. Production of drugs as nanosuspensions can be developed for drug delivery systems as an oral formulation and nonoral administration. Here, this review describes the methods of pharmaceutical nanosuspension production including advantages and disadvantages, potential benefits, characterization tests, and pharmaceutical applications in drug delivery.

  12. Increasing orUnderselling ?

    Institute of Scientific and Technical Information of China (English)

    Guo Yan

    2011-01-01

    The downgrade of sovereignty credit rating of the United States causes new concerns on US treasury bonds.According to the report issued recently by Treasury Ministry of United States,China as the largest oversea holding country of US treasury bonds continues to buy US treasury bonds,with an increase of 5.7 billion US dollars in June,marking the third month in line.Till now,China has US treasury bonds up to a new record of 1,165.5 billion US dollars,which amounts to 26% of the oversea holding US treasury bonds,based on market estimation,followed by Japan (20%),UK (8%),Brazil (5%)and Russia (3%).However,in sharp contrast to China,other countries are underselling us treasury bonds,particularly Russia.Data shows that since October 2010,Russia has been selling US treasury bonds for eight months in line,up to 38% of their holdings.Now Russia only holds 111 billion US dollars of US treasury bond.Why the countries take totally different attitudes towards to US treasury bonds? Tne reporter interviewed the experts from China and Russia for more details.

  13. Increasing instruction time in school does increase learning

    DEFF Research Database (Denmark)

    Humlum, Maria; Nandrup, Anne Brink; Andersen, Simon Calmar

    2016-01-01

    Increasing instruction time in school is a central element in the attempts of many governments to improve student learning, but prior research—mainly based on observational data—disputes the effect of this approach and points out the potential negative effects on student behavior. Based on a large......-scale, cluster-randomized trial, we find that increasing instruction time increases student learning and that a general increase in instruction time is at least as efficient as an expert-developed, detailed teaching program that increases instruction with the same amount of time. These findings support the value...

  14. Strong increase of solar panels

    International Nuclear Information System (INIS)

    The number of installed solar panels in 2011 has increased again. 40 megawatt of new panels have been installed. This increase is twice as high as the year before. The production of solar power increased to 90 million kWh in 2011 as a result of this expansion. However, the share of solar power in total energy use is still very limited.

  15. Increasing tobacco taxes : A cheap tool to increase public health

    NARCIS (Netherlands)

    van Baal, Pieter H. M.; Brouwer, Werner B. F.; Hoogenveen, Rudolf T.; Feenstra, Talitha L.

    2007-01-01

    Introduction: Several studies have estimated health effects resulting from tobacco tax increases. However, studies on the cost effectiveness of tobacco taxes are scarce. The aim of this study was to estimate the cost effectiveness of tobacco tax increases from a health care perspective, explicitly c

  16. Assessing and Increasing Physical Activity

    Science.gov (United States)

    Van Camp, Carole M.; Hayes, Lynda B.

    2012-01-01

    Increasing physical activity is a crucial component of any comprehensive approach to combat the growing obesity epidemic. This review summarizes recent behavioral research on the measurement of physical activity and interventions aimed at increasing physical activity and provides directions for future research.

  17. RESTAURANT NO. 2: PRICE INCREASES

    CERN Multimedia

    2003-01-01

    'DSR', the concession holder of Restaurant no. 2 (bldg. 504 - Meyrin site), has submitted to the Restaurant Supervisory Committee a request to increase certain prices. After close examination, the Committee has established that the proposed increases are compatible with the relevant price indexing mechanisms and other contractual conditions. The new prices will apply as from Monday, June 30, 2003.

  18. RESTAURANT NO. 3: PRICE INCREASES

    CERN Multimedia

    2003-01-01

    'AVENANCE', the concession holder of Restaurant no. 3 (bldg. 866 - Prévessin site), has submitted to the Restaurant Supervisory Committee a request to increase its prices. After close examination, the Committee has established that the proposed increases are compatible with the relevant price indexing mechanisms and other contractual conditions. The new prices will apply as from Monday, June 2, 2003.

  19. Strategies to Increase Physical Activity

    OpenAIRE

    Tuso, Phillip

    2015-01-01

    One key intervention to prevent preventable diseases and to make health care more affordable is to increase the percentage of Americans who are physically active. No single intervention will increase activity rates, but a group of interventions working together in synergy may be the stimulus needed to get Americans moving.

  20. Demand for Phenol Increases Steadily

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    @@ Dramatic growth in output By the end of 2006 China's capacity to produce phenol had reached more than 600 thousand t/a. The total output was 523 thousand tons in 2006,an increase of 18.1% over 2005.

  1. Demand for SBR Increased Steadily

    Institute of Scientific and Technical Information of China (English)

    Gao Yiwen

    2007-01-01

    @@ 1 Rapid consumption growth Consumption of styrene-butadiene rubber (SBR) in China was 747 thousand tons in 2006, an increase of 21% over 2005, with faster growth than the average rubber consumption growth.

  2. Increasing Learning from TV News.

    Science.gov (United States)

    Perloff, Richard M.; And Others

    1982-01-01

    Describes an experiment that manipulated two variables, repetition and pausing for viewer "digestion" of information in a news telecast. Concludes that the use of repetition increased viewers' retention of information, but that pauses did not. (FL)

  3. RESTAURANT NO. 3 : PRICE INCREASES

    CERN Multimedia

    Restaurant Supervisory Committee

    2002-01-01

    'AVENANCE', the concession holder of Restaurant no. 3 (bldg. 866 - Prévessin site), has submitted to the Restaurant Supervisory Committee a request to increase its prices. After close examination, the Committee has established that the proposed increases are compatible with the relevant price indexing mechanisms and other contractual conditions. The new prices will apply as from Monday, June 3, 2002. Restaurant Supervisory Committee, tel. 77551

  4. INCREASING RETURN TO SMART CITIES

    OpenAIRE

    Lööf, Hans; Nabavi, Pardis

    2012-01-01

    Increased urbanization, global warming and sustainable growth belong to the major contemporary policy challenges. Today cities are home to more than 50% of the world population, the largest 600 urban centers generate about 60% of global GDP, and the agglomerated areas are responsible for 75% of world carbon emissions. The UN estimates that 70% of the world's growing population will live in cities by 2050. At the same time the world population is expected to increase from 7 billion people to 9...

  5. Research Evaluation to Increase Impact

    OpenAIRE

    Templeton, Deborah J.

    2003-01-01

    Research evaluation, undertaken throughout the life of the research activity, is an important tool for research managers. When correctly implemented, research evaluation can help to increase the success of existing and future research projects and programs, thereby increasing the social, economic and/or environmental impact of the research. The relationship between the life-cycle of the research project and the type of evaluation determines the appropriate type of evaluation to be undertaken....

  6. Meclofenamate increases ventilation in lambs.

    Science.gov (United States)

    Guerra, F A; Savich, R D; Clyman, R I; Kitterman, J A

    1989-01-01

    To investigate the effects of the prostaglandin synthetase inhibitor, meclofenamate, on postnatal ventilation, we studied 11 unanaesthetised, spontaneously-breathing lambs at an average age of 7.9 +/- 1.1 days (SEM; range 5-14 days) and an average weight of 4.9 +/- 0.5 kg (range 3.0-7.0 kg). After a 30-min control period we infused 4.23 mg/kg meclofenamate over 10 min and then gave 0.23 mg/h per kg for the remainder of the 4 h. Ventilation increased progressively from a control value of 515 +/- 72 ml/min per kg to a maximum of 753 +/- 100 ml/min per kg after 3h of infusion (P less than 0.05) due to an increased breathing rate; the effects were similar during both high- and low-voltage electrocortical activity. There were no significant changes in tidal volume, heart rate, blood pressure, arterial pH or PaCO2, the increased ventilation resulted from either an increase in dead space ventilation or an increase in CO2 production. This study indicates that meclofenamate causes an increase in ventilation in lambs but no changes in pH of PaCO2. The mechanism and site of action remain to be defined. PMID:2507622

  7. Increasing work-time influence

    DEFF Research Database (Denmark)

    Nabe-Nielsen, Kirsten; Garde, Anne Helene; Aust, Birgit;

    2012-01-01

    This quasi-experimental study investigated how an intervention aiming at increasing eldercare workers' influence on their working hours affected the flexibility, variability, regularity and predictability of the working hours. We used baseline (n = 296) and follow-up (n = 274) questionnaire data......-operation with colleagues. In subgroup B and C, the participants ended up discussing the potential consequences of more work-time influence without actually implementing any changes. PRACTITIONER SUMMARY: Employee work-time influence may buffer the adverse effects of shift work. However, our intervention study suggested...... that while increasing the individual flexibility, increasing work-time influence may also result in decreased regularity of the working hours and less continuity in the care of clients and co-operation with colleagues....

  8. Increased burnup of fuel elements

    International Nuclear Information System (INIS)

    The specialists' group for fuel elements of the Kerntechnische Gesellschaft e.V. held a meeting on ''Increased Burnup of Fuel Elements'' on 9th and 10th of November 1982 at the GKSS Research Center Geesthacht. Most papers dealt with the problems of burnup increase of fuel elements for light water reactors with respect to fuel manufacturing, power plant operation and reprocessing. Review papers were given on the burnup limits for high temperature gas cooled reactors and sodium fast breeder reactors. The meeting ended with a presentation of the technical equipment of the hot laboratory of the GKSS and the programs which are in progress there. (orig.)

  9. Increased Spreading Activation in Depression

    Science.gov (United States)

    Foster, Paul S.; Yung, Raegan C.; Branch, Kaylei K.; Stringer, Kristi; Ferguson, Brad J.; Sullivan, William; Drago, Valeria

    2011-01-01

    The dopaminergic system is implicated in depressive disorders and research has also shown that dopamine constricts lexical/semantic networks by reducing spreading activation. Hence, depression, which is linked to reductions of dopamine, may be associated with increased spreading activation. However, research has generally found no effects of…

  10. Electromigration early resistance increase measurements

    NARCIS (Netherlands)

    Niehof, J.; Flinn, P.A.; Maloney, T.J.

    1993-01-01

    An early resistance change measurement set-up, using an AC bridge technique, has been developed, and measurements have been performed. Large sample-to-sample variations occur. The characteristic time for the resistance change curve is shorter for resistance increase (under current stress) than for r

  11. Protistas "incerta sedis" de ofídios do Brasil: Toddia França, 1912 e Pirhemocyton Chatton & Blanc, 1914: estudo comparativo "Incerta sedis" Portists from Brazilian snakes: Toddia França, 1912 and Pirhemocyton Chatton & Blanc, 1914: comparative study

    Directory of Open Access Journals (Sweden)

    Maria Auxiliadora de Sousa

    1973-01-01

    Full Text Available Dentre 683 serpentes procedentes de diferentes pontos do Brasil, 16 (2,4% apresentaram em seu sangue (em glóbulos vermelhos maturos e imaturos Toddia ou Pirhemocyton. Denominamos provisoriamente Toddia os parasitos encontrados em Bothrops moojeni, B. pradoi, B. jararaca e Chironius flavolineatus e Pirhemocyton os encontrados em B. alternatus; fornecemos a primeira referência nestes hospedeiros. Estes parasitos foram estudados comparativamente com o objetivo de estabelecermos a extensão de suas afinidades. Com Toddia de B. moojeni realizamos técnica citoquímica para caracterização de ácidos nucleicos e encontramos positividade apenas para DNA. As maiores afinidades entre os parasitos estudados residiam nas características dos corpúsculos cromáticos e alterações celulares que determinam; observarmos quase total identidade nas maneiras como os parasitos da mesma espécie de serpente se apresentaram. Constatamos a coexistência de inclusões globóides, cristalóides e formas intermediárias associada aos parasitos de B. pradoi e B. alternatus; em C. flavolineatus encotnramos além dos cristais, raríssimas formas intermediárias entre estes e globóides. Afinidades entre Toddia e Pirhemocyton abrangendo o tipo de inclusão contrariam as descrições originais, tornando pouco válida esta separação genérica. Discutimos também os fundamentos da criação das espécies e a possível natureza virótica destes parasitos.Among 683 snakes from several localities of Brazil, 16 (2,4% presented in their blood (in mature or immature red cells Toddia or Pirhemocyton. We named provisorily Toddia the parasites found in Bothrops moojeni, B. pradoi, B. jararaca and Chironius flavolineatus and Pirhemocyton those found in B. alternatus; we give the first reference in these hosts. These parasites were comparatively studied in view to establish the range of their relationship. With Toddia from B. moojeni we attempted nucleic acids by cytochemical

  12. Molecular cloning of a hyaluronidase from Bothrops pauloensis venom gland

    OpenAIRE

    Castanheira, Letícia Eulalio; Rodrigues, Renata Santos; Boldrini-França, Johara; Fonseca, Fernando PP; Henrique-Silva, Flávio; Homsi-Brandeburgo, Maria I; Rodrigues, Veridiana M.

    2014-01-01

    Background Hyaluronate is one of the major components of extracellular matrix from vertebrates whose breakdown is catalyzed by the enzyme hyaluronidase. These enzymes are widely described in snake venoms, in which they facilitate the spreading of the main toxins in the victim’s body during the envenoming. Snake venoms also present some variants (hyaluronidases-like substances) that are probably originated by alternative splicing, even though their relevance in envenomation is still under inve...

  13. Clinical and immunological aspects of envenomations by Bothrops snakes

    OpenAIRE

    KPO Luna; MB da Silva; VRA Pereira

    2011-01-01

    Accidents caused by snakes, especially in tropical and subtropical countries, still constitute a serious public health problem due to the lack of knowledge of health professionals and the precariousness of health systems in the regions where most accidents occur. Snake venoms contain a range of molecules that may provoke local swelling, pain, renal and respiratory insufficiencies. The study of the effects of each molecule on humans can help the development of complementary therapy. Similarly,...

  14. How to increase living donation.

    Science.gov (United States)

    Davis, Connie L

    2011-04-01

    Living donation is the key to increasing access to successful solid organ transplantation worldwide. However, the means to expanding the number of living donors on a global scale are not known. Although there have been many suggestions for the best approach, cultural issues may limit the effectiveness of some strategies. Only a few ideas have been studied, and one in particular- outright payment to donors - may raise ethical issues that are difficult to surmount and might negatively alter altruistic behavior. With respect to the present environment, this article will describe some of the approaches that are being discussed to increase the number of living donors, with a particular focus on kidney transplantation. PMID:21210867

  15. Increasing Accuracy in Environmental Measurements

    Science.gov (United States)

    Jacksier, Tracey; Fernandes, Adelino; Matthew, Matt; Lehmann, Horst

    2016-04-01

    Human activity is increasing the concentrations of green house gases (GHG) in the atmosphere which results in temperature increases. High precision is a key requirement of atmospheric measurements to study the global carbon cycle and its effect on climate change. Natural air containing stable isotopes are used in GHG monitoring to calibrate analytical equipment. This presentation will examine the natural air and isotopic mixture preparation process, for both molecular and isotopic concentrations, for a range of components and delta values. The role of precisely characterized source material will be presented. Analysis of individual cylinders within multiple batches will be presented to demonstrate the ability to dynamically fill multiple cylinders containing identical compositions without isotopic fractionation. Additional emphasis will focus on the ability to adjust isotope ratios to more closely bracket sample types without the reliance on combusting naturally occurring materials, thereby improving analytical accuracy.

  16. Price Increases from Online Privacy

    OpenAIRE

    Ward, Michael R; Chen, Yu-Ching

    2001-01-01

    Consumers value keeping some information about them private from potential marketers. E-commerce dramatically increases the potential for marketers to accumulate otherwise private information about potential customers. Online marketers claim that this information enables them to better market their products. Policy makers are currently drafting rules to regulate the way in which these marketers can collect, store, and share this information. However, there is little evidence yet either of con...

  17. Production Increased with Price Rise

    Institute of Scientific and Technical Information of China (English)

    Feng Shiliang

    2007-01-01

    @@ In April 2007, China's petroleum and chemical industry remained up trend in production. Among the 65 types of petrochemical products that have been tracked by CPCIA (China Petroleum and Chemical Industry Association), the output of 61 types increased compared with April 2006,accounting for 93.8% of the total, and the output of 15 types, in particular, grew by over 20%, accounting for 23.1%.

  18. NSF and NASA budgets increased

    Science.gov (United States)

    Bell, Peter M.

    Research budgets of several of the federal government agencies were increased significantly over the Reagan administration's requests in the House of Representative's appropriations bill H.R. 4034. These budgets had been removed from the Reagan administration's omnibus reconciliation bill, and thus there were worries expressed that certain research funding could be in jeopardy. The rationale was that because the requests were voted on individually on the floor of the House, many sections of the budgets would be subjected to extra scrutiny, which would lead to more cuts.The National Science Foundation (NSF) budget request had been cut and reordered by the Office of Management and Budget (OMB) by making sharp reductions in programs of the social sciences and in programs of science and engineering education. There were fears that these programs would be reinstated to the original request level, at the expense of the budgets of other research activities. These fears materialized, but only momentarily. Efforts to cut the research activities by the House Appropriations Committee were soundly defeated. The budget was supported, with additional increases to provide for the education programs, by a high margin, which included most Republican and Democratic members of the House of Representatives. The overall NSF budget, as passed, has a total appropriation of $1103.5 million, compared with the Administration's request of $1033.5 million (the Fiscal Year 1981 appropriation for the NSF was $1022.4 million). The House approved budget included increases of $44.9 million in research and $25.1 million in science and engineering education. Included in the research budget increase were recommendations by the House Appropriations Committee for support of the social sciences and for the international affairs programs. Also included in the recommendations was support of interdisciplinary research programs that cut across the directorates of the NSF.

  19. Production Increased with Higher Prices

    Institute of Scientific and Technical Information of China (English)

    Feng shiling

    2007-01-01

    @@ In the first quarter of 2007, China's petroleum and chemical industry remained up trend in production.Among the 66 types of petrochemical products that have been tracked by CPCIA (China Petroleum and Chemical Industry Association), the output of 58, types increased compared with the same period of 2006, accounting for 87.9% of the total, and the output of 19 types, in particular, grew by over20%, accounting for 28.8%.

  20. How trade unions increase welfare

    OpenAIRE

    Donado, Alejandro; Klaus WALDE

    2010-01-01

    Historically, worker movements have played a crucial role in making workplaces safer. Firms traditionally oppose better health standards. According to our interpretation, workplace safety is costly for firms but increases the average health of workers and thereby the aggregate labour supply. A laissez-faire approach in which firms set safety standards is suboptimal as workers are not fully informed of health risks associated with jobs. Safety standards set by better-informed trade unions are ...

  1. High Heels Increase Women's Attractiveness.

    Science.gov (United States)

    Guéguen, Nicolas

    2015-11-01

    Research has found that the appearance of women's apparel helps increase their attractiveness as rated by men and that men care more about physical features in potential opposite-sex mates. However, the effect of sartorial appearance has received little interest from scientists. In a series of studies, the length of women's shoe heels was examined. A woman confederate wearing black shoes with 0, 5, or 9 cm heels asked men for help in various circumstances. In Study 1, she asked men to respond to a short survey on gender equality. In Study 2, the confederate asked men and women to participate in a survey on local food habit consumption. In Study 3, men and women in the street were observed while walking in back of the female confederate who dropped a glove apparently unaware of her loss. It was found that men's helping behavior increased as soon as heel length increased. However, heel length had no effect on women's helping behavior. It was also found that men spontaneously approached women more quickly when they wore high-heeled shoes (Study 4). Change in gait, foot-size judgment, and misattribution of sexiness and sexual intent were used as possible explanations. PMID:25408499

  2. Increase Productivity Through Knowledge Management

    Science.gov (United States)

    Gavrikova, N. A.; Dolgih, I. N.; Dyrina, E. N.

    2016-04-01

    Increase in competition level requires companies to improve the efficiency of work force use characterized by labor productivity. Professional knowledge of staff and its experience play the key role in it. The results of Extrusion Line operator’s working time analysis are performed in this article. The analysis revealed that the reasons of working time ineffective use connected with inadequate information exchange and knowledge management in the company. Authors suggest the way to solve this problem: the main sources of knowledge in engineering enterprise have been defined, the conditions of success and the stages of knowledge management control have been stated.

  3. Do trade missions increase trade?

    OpenAIRE

    Head, Keith; Ries, John

    2010-01-01

    In an effort to stimulate trade, Canada has conducted regular trade missions starting in 1994, often led by the Prime Minister. According to the Canadian government, these missions generated tens of billions of dollars in new business deals. This paper uses bilateral trade data to assess this claim. We find that Canada exports and imports above-normal amounts to the countries to which it sent trade missions. However, the missions do not seem to have caused an increase in trade. In the preferr...

  4. Increasing Security in Cloud Environment

    OpenAIRE

    Naik, Priyanka; Sanyal, Sugata

    2013-01-01

    The concept of cloud computing was introduced to meet the increase in demand for new application for a project, and to provide a large storage facility whenever or wherever a user needs it. The cloud system facility helped many industries as well as individual users to get authentic software at a very low cost. But with this new system comes the major concern of security, as the connection to the cloud is through the web and the data and application availability need to be handled for each cl...

  5. [Ozone decline and UV increase].

    Science.gov (United States)

    Winkler, P; Trepte, S

    2004-02-01

    The following results have been obtained from long-term observations on the ozone layer and UV at the Meteorological Observatory Hohenpeigenberg:The seasonally varying decline of the ozone layer determines the maximum exposure to UV. Since ozone decline shows the highest rates in the spring months the UV exposure has most strongly increased in this time of the year. This is especially important because in spring the human skin is not adapted to UV exposure. Weather changes from day to day can induce rapid ozone reductions in spring about -30% which in turn is followed by an increase in UV of about 40%. Clouds, especially the transparent cirrus clouds (high clouds consisting of ice particles) have increased in frequency during spring and fall while a decrease is observed in summer. This change in cloudiness reduces the daily UV dose in spring and fall while it is enhanced in summer. With increasing height above sea level UV rises by roughly 10% per 1000 m (rule of thumb). Snow reflects the UV-radiation by up to 80% enhancing the UV-doses at relevant conditions. Strong volcano eruptions destroy ozone in the stratosphere additionally during 1-2 years after the eruption. Therafter the ozone layer recovers. In April 1993, after the eruption of Mt. Pinatubo (1991), the UV burden was still 40% higher than average. Miniholes and streamers can appear unexpected on a short-time scale and cross over Central Europe within 1-2 days, thus enhancing UV irradiation. The human skin reacts to UV exposure depending on the type of skin. The campaign "Sonne(n) mit Verstand" of the Bavarian Ministries for Environment, for Health and for Education informs about the danger of UV radiation (see www.sonne-mit-ver-stand.de). The German Weather Service informs the public on present developments of the ozone layer and relevant topics byits ozone bulletin, which is also available via internet under (www.dwd.de/deFundE/Observator/MOHp/hp2/ozon/bulletin.htm). PMID:14770335

  6. Increasing situational awareness using smartphones

    Science.gov (United States)

    Boddhu, Sanjay K.; Williams, Robert L.; Wasser, Edward; Kode, Niranjan

    2012-06-01

    In recent years, the United States Armed Services and various law enforcement agencies have shown increasing interest in evaluating the feasibility of using smartphones and hand-held devices as part of the standard gear for its personnel, who are actively engaged on battlefield or in crime-prone areas. The primary motive driving analysis efforts to employ smartphone-based technologies is the prospect of the increased "Situational Awareness" achievable thru a digitally connected network of armed personnel. Personnel would be equipped with customized smart applications that use the device's sensors (GPS, camera, compass, etc...) to sense the hostile environments as well as enabling them to perform collaborative tasks to effectively complete a given mission. In this vein, as part of the Summer At The Edge (SATE) program, a group of student interns under the guidance of mentors from Qbase and AFRL, have employed smartphones and built three smart applications to tackle three real-world scenarios: PinPoint, IStream, and Cooperative GPS. This paper provides implementation details for these prototype applications, along with the supporting visualization and sensor cloud platforms and discusses results obtained from field testing of the same. Further, the paper concludes by providing the implications of the present work and insights into future work.

  7. [Patients' increasing role in healthcare].

    Science.gov (United States)

    Colombo, Cinzia

    2016-03-01

    Since the late '90s citizens have been increasingly involved in healthcare, thanks to a shift in access to healthcare information, greater patients' and citizens' awareness about their rights and needs, and a change in physicians' attitudes and behavior. At the same time, to ensure the sustainability of the national health services, patients' needs are increasingly being examined in relation to the services and interventions needed, and to foster informed demand by patients and citizens, and appropriate prescriptions by physicians. Nowadays, patients already have a section in an authoritative medical journal. Working with clinicians, they set research priorities and the outcomes to be studied. Especially in UK and USA, they are invited to participate in the design and development of trials and in deciding which trials should be funded. The situation varies widely in different countries, though involving citizens in healthcare decisions is a common mantra. Even when they consult a clinician, the patients' role has evolved. People ask for information more often and want an active part in decisions about their health. Physicians start to pay more attention to the person and the social context, feelings and emotions, and person-centered care has become a reference in the doctor-patient relationship. This article offers an overview of the changing roles of patients and citizens in healthcare.

  8. Thunderstorms Increase Mercury Wet Deposition.

    Science.gov (United States)

    Holmes, Christopher D; Krishnamurthy, Nishanth P; Caffrey, Jane M; Landing, William M; Edgerton, Eric S; Knapp, Kenneth R; Nair, Udaysankar S

    2016-09-01

    Mercury (Hg) wet deposition, transfer from the atmosphere to Earth's surface by precipitation, in the United States is highest in locations and seasons with frequent deep convective thunderstorms, but it has never been demonstrated whether the connection is causal or simple coincidence. We use rainwater samples from over 800 individual precipitation events to show that thunderstorms increase Hg concentrations by 50% relative to weak convective or stratiform events of equal precipitation depth. Radar and satellite observations reveal that strong convection reaching the upper troposphere (where high atmospheric concentrations of soluble, oxidized mercury species (Hg(II)) are known to reside) produces the highest Hg concentrations in rain. As a result, precipitation meteorology, especially thunderstorm frequency and total rainfall, explains differences in Hg deposition between study sites located in the eastern United States. Assessing the fate of atmospheric mercury thus requires bridging the scales of global transport and convective precipitation. PMID:27464305

  9. Increasing of Reliability of Converter

    Directory of Open Access Journals (Sweden)

    Trebuňa, F.

    2006-01-01

    Full Text Available The innovative activities in steel production invoke exploitation of new machines with the higher production capacity and productivity of labor. The paper deals with deformation and stress analysis of carrying parts of converter pedestal on the base of which the proposals and supporting measures were made that had the aim to increase reliability of the converter during steel production. This was achieved by lifespan prolongation of anchor and connecting bolts of converter pedestal, by change of stiffness of connected elements as well as by correction of nuts of bolted connections. The realization of structural changes decreased loading amplitudes and consequently the vibrations of pedestal. Solution was verified by numerical and experimental procedures of mechanics.

  10. Propofol directly increases tau phosphorylation.

    Directory of Open Access Journals (Sweden)

    Robert A Whittington

    Full Text Available In Alzheimer's disease (AD and other tauopathies, the microtubule-associated protein tau can undergo aberrant hyperphosphorylation potentially leading to the development of neurofibrillary pathology. Anesthetics have been previously shown to induce tau hyperphosphorylation through a mechanism involving hypothermia-induced inhibition of protein phosphatase 2A (PP2A activity. However, the effects of propofol, a common clinically used intravenous anesthetic, on tau phosphorylation under normothermic conditions are unknown. We investigated the effects of a general anesthetic dose of propofol on levels of phosphorylated tau in the mouse hippocampus and cortex under normothermic conditions. Thirty min following the administration of propofol 250 mg/kg i.p., significant increases in tau phosphorylation were observed at the AT8, CP13, and PHF-1 phosphoepitopes in the hippocampus, as well as at AT8, PHF-1, MC6, pS262, and pS422 epitopes in the cortex. However, we did not detect somatodendritic relocalization of tau. In both brain regions, tau hyperphosphorylation persisted at the AT8 epitope 2 h following propofol, although the sedative effects of the drug were no longer evident at this time point. By 6 h following propofol, levels of phosphorylated tau at AT8 returned to control levels. An initial decrease in the activity and expression of PP2A were observed, suggesting that PP2A inhibition is at least partly responsible for the hyperphosphorylation of tau at multiple sites following 30 min of propofol exposure. We also examined tau phosphorylation in SH-SY5Y cells transfected to overexpress human tau. A 1 h exposure to a clinically relevant concentration of propofol in vitro was also associated with tau hyperphosphorylation. These findings suggest that propofol increases tau phosphorylation both in vivo and in vitro under normothermic conditions, and further studies are warranted to determine the impact of this anesthetic on the acceleration of

  11. Slow motion increases perceived intent.

    Science.gov (United States)

    Caruso, Eugene M; Burns, Zachary C; Converse, Benjamin A

    2016-08-16

    To determine the appropriate punishment for a harmful action, people must often make inferences about the transgressor's intent. In courtrooms and popular media, such inferences increasingly rely on video evidence, which is often played in "slow motion." Four experiments (n = 1,610) involving real surveillance footage from a murder or broadcast replays of violent contact in professional football demonstrate that viewing an action in slow motion, compared with regular speed, can cause viewers to perceive an action as more intentional. This slow motion intentionality bias occurred, in part, because slow motion video caused participants to feel like the actor had more time to act, even when they knew how much clock time had actually elapsed. Four additional experiments (n = 2,737) reveal that allowing viewers to see both regular speed and slow motion replay mitigates the bias, but does not eliminate it. We conclude that an empirical understanding of the effect of slow motion on mental state attribution should inform the life-or-death decisions that are currently based on tacit assumptions about the objectivity of human perception. PMID:27482091

  12. Vocal attractiveness increases by averaging.

    Science.gov (United States)

    Bruckert, Laetitia; Bestelmeyer, Patricia; Latinus, Marianne; Rouger, Julien; Charest, Ian; Rousselet, Guillaume A; Kawahara, Hideki; Belin, Pascal

    2010-01-26

    Vocal attractiveness has a profound influence on listeners-a bias known as the "what sounds beautiful is good" vocal attractiveness stereotype [1]-with tangible impact on a voice owner's success at mating, job applications, and/or elections. The prevailing view holds that attractive voices are those that signal desirable attributes in a potential mate [2-4]-e.g., lower pitch in male voices. However, this account does not explain our preferences in more general social contexts in which voices of both genders are evaluated. Here we show that averaging voices via auditory morphing [5] results in more attractive voices, irrespective of the speaker's or listener's gender. Moreover, we show that this phenomenon is largely explained by two independent by-products of averaging: a smoother voice texture (reduced aperiodicities) and a greater similarity in pitch and timbre with the average of all voices (reduced "distance to mean"). These results provide the first evidence for a phenomenon of vocal attractiveness increases by averaging, analogous to a well-established effect of facial averaging [6, 7]. They highlight prototype-based coding [8] as a central feature of voice perception, emphasizing the similarity in the mechanisms of face and voice perception.

  13. Vocal attractiveness increases by averaging.

    Science.gov (United States)

    Bruckert, Laetitia; Bestelmeyer, Patricia; Latinus, Marianne; Rouger, Julien; Charest, Ian; Rousselet, Guillaume A; Kawahara, Hideki; Belin, Pascal

    2010-01-26

    Vocal attractiveness has a profound influence on listeners-a bias known as the "what sounds beautiful is good" vocal attractiveness stereotype [1]-with tangible impact on a voice owner's success at mating, job applications, and/or elections. The prevailing view holds that attractive voices are those that signal desirable attributes in a potential mate [2-4]-e.g., lower pitch in male voices. However, this account does not explain our preferences in more general social contexts in which voices of both genders are evaluated. Here we show that averaging voices via auditory morphing [5] results in more attractive voices, irrespective of the speaker's or listener's gender. Moreover, we show that this phenomenon is largely explained by two independent by-products of averaging: a smoother voice texture (reduced aperiodicities) and a greater similarity in pitch and timbre with the average of all voices (reduced "distance to mean"). These results provide the first evidence for a phenomenon of vocal attractiveness increases by averaging, analogous to a well-established effect of facial averaging [6, 7]. They highlight prototype-based coding [8] as a central feature of voice perception, emphasizing the similarity in the mechanisms of face and voice perception. PMID:20129047

  14. The increasing burden of depression

    Directory of Open Access Journals (Sweden)

    Lépine J-P

    2011-05-01

    Full Text Available Jean-Pierre Lépine1, Mike Briley21Hôpital Lariboisière Fernand Widal, Assistance Publique Hôpitaux de Paris Unité INSERM 705 CNRS UMR 8206, Université Paris Diderot, Paris, France; 2NeuroBiz Consulting and Communication, Castres, FranceAbstract: Recent epidemiological surveys conducted in general populations have found that the lifetime prevalence of depression is in the range of 10% to 15%. Mood disorders, as defined by the World Mental Health and the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, have a 12-month prevalence which varies from 3% in Japan to over 9% in the US. A recent American survey found the prevalence of current depression to be 9% and the rate of current major depression to be 3.4%. All studies of depressive disorders have stressed the importance of the mortality and morbidity associated with depression. The mortality risk for suicide in depressed patients is more than 20-fold greater than in the general population. Recent studies have also shown the importance of depression as a risk factor for cardiovascular death. The risk of cardiac mortality after an initial myocardial infarction is greater in patients with depression and related to the severity of the depressive episode. Greater severity of depressive symptoms has been found to be associated with significantly higher risk of all-cause mortality including cardiovascular death and stroke. In addition to mortality, functional impairment and disability associated with depression have been consistently reported. Depression increases the risk of decreased workplace productivity and absenteeism resulting in lowered income or unemployment. Absenteeism and presenteeism (being physically present at work but functioning suboptimally have been estimated to result in a loss of $36.6 billion per year in the US. Worldwide projections by the World Health Organization for the year 2030 identify unipolar major depression as the leading cause of disease burden

  15. Increased VLDL in nephrotic patients results from a decreased catabolism while increased LDL results from increased synthesis

    NARCIS (Netherlands)

    de Sain-van der Velden, M; Kaysen, GA; Barrett, HA; Stellaard, F; Gadellaa, MM; Voorbij, HA; Reijngoud, DJ; Rabelink, TJ

    1998-01-01

    Increased very low density lipoprotein (VLDL) in nephrotic patients results from a decreased catabolism while increased low density lipoprotein (LDL) results from increased synthesis. Hyperlipidemias a hallmark of nephrotic syndrome that has been associated with increased risk for ischemic heart dis

  16. Increased skeletal muscle capillarization enhances insulin sensitivity

    DEFF Research Database (Denmark)

    Åkerström, Thorbjörn; Laub, Lasse; Vedel, Kenneth;

    2014-01-01

    Increased skeletal muscle capillarization is associated with improved glucose tolerance and insulin sensitivity. However, a possible causal relationship has not previously been identified. We therefore investigated whether increased skeletal muscle capillarization increases insulin sensitivity......-body insulin sensitivity increased by ~24% and insulin-stimulated skeletal muscle 2-deoxy-[(3)H]-Glucose disposal increased by ~30% concomitant with a ~20% increase in skeletal muscle capillarization. Adipose tissue insulin sensitivity was not affected by the treatment. Insulin-stimulated muscle glucose uptake...... the rats on any other parameters measured. We conclude that an increase in skeletal muscle capillarization is associated with increased insulin sensitivity. These data point towards the importance of increasing skeletal muscle capillarization for prevention or treatment of type 2 diabetes....

  17. 3 Ways to Increase Positive Emotions

    Science.gov (United States)

    ... Friend Who Cuts? 3 Ways to Increase Positive Emotions KidsHealth > For Teens > 3 Ways to Increase Positive ... to give yourself a boost. Track Your Positive Emotions Name the positive emotions you're already familiar ...

  18. Oral microbiota of Brazilian captive snakes

    Directory of Open Access Journals (Sweden)

    MG Fonseca

    2009-01-01

    Full Text Available The present work aimed to determine the oral microbiotic composition of snakes from São José do Rio Preto city, São Paulo State, Brazil. Ten snake species, comprising the families Boidae, Colubridae, Elapidae and Viperidae, were submitted to microbiological examination of their oral cavity, which indicated positivity for all buccal samples. Gram-negative bacilli, gram-negative cocci bacilli, gram-positive bacilli and gram-positive cocci were isolated from the snakes. Among isolated bacterium species, the occurrence of coagulase-negative staphylococci in the buccal cavity of Crotalus durissus (Viperiade, Eunectes murinus (Boidae, Mastigodryas bifossatus (Colubridae and Bacillus subtilis, common to oral cavity of Bothrops alternatus (Viperidae and Phalotris mertensi (Colubridae, was detected. It was observed higher diversity of isolated bacteria from the oral cavity of Micrurus frontalis (Elapidae and Philodryas nattereri (Colubridae, as well as the prevalence of gram-positive baccillus and gram-positive cocci. The composition of the oral microbiota of the studied snakes, with or without inoculating fangs, is diverse and also related to the formation of abscesses at the bite site in the victims of the ophidian accidents, and to pathogenic processes in the snakes that host these microorganisms.

  19. Inhibition of mutalysin II, a metalloproteinase from bushmaster snake venom by human alpha2-macroglobulin and rabbit immunoglobulin.

    Science.gov (United States)

    Souza, C T; Moura, M B; Magalhaes, A; Heneine, L G; Olortegui, C C; Diniz, C R; Sanchez, E F

    2001-09-01

    Mutalysin II is a 22.5-kDa zinc endopeptidase isolated from Lachesis muta muta snake venom. In order to determine whether the inhibitors human alpha2-macroglobulin (alpha2-M) and rabbit antibody to mutalysin II share a common mechanism, we have investigated the inhibition of mutalysin II by these two different glycoproteins. The proteolytic activity of mutalysin II with dimethylcasein as substrate was completely inhibited by human alpha2-M and by a purified rabbit antibody to mutalysin II. The protection of fibrin(ogen) digestion by alpha2-M was slightly better than the protection offered by the antibody. In addition, the purified antibody reacted only with the metalloproteinase in bushmaster venom, as demonstrated by immunodiffusion. SDS-PAGE analysis of reduced samples showed that the interaction of mutalysin II with alpha2-M resulted in the formation of high molecular complex ( approximately 180000) and M(r) 90000 fragments generated by the venom enzyme. Also, fragments at 85 and 23 kDa were detected under non-reducing conditions after incubation of rabbit immunoglobulin with enzyme. Proteolysis of dimethylcasein as substrate revealed that the stoichiometry of inhibition was 1.0 mol of human alpha2-M and 1.5 mol of rabbit IgG antimutalysin II per mole of enzyme. Furthermore, dimethylcasein hydrolysis indicated that several viperid snake venoms, including Bothrops atrox, B. alternatus and Trimeresurus flavoviridis cross-reacted with the specific rabbit antibody to varying degrees. PMID:11544086

  20. Increasing ideological tolerance in social psychology.

    Science.gov (United States)

    Inbar, Yoel; Lammers, Joris

    2015-01-01

    We argue that recognizing current ideological diversity in social psychology and promoting tolerance of minority views is just as important as increasing the number of non-liberal researchers. Increasing tolerance will allow individuals in the minority to express dissenting views, which will improve psychological science by reducing bias. We present four recommendations for increasing tolerance.

  1. A importância dos acidentes ofídicos como causa de mortes em bovinos no Brasil The importance of snake bites as cause of cattle death in Brazil

    Directory of Open Access Journals (Sweden)

    Carlos Hubinger Tokarnia

    2006-06-01

    Full Text Available A revisão da literatura pertinente indica que as opiniões sobre a importância dos acidentes ofídicos, como causa de mortes em bovinos no Brasil, são divergentes no meio veterinário. Enquanto alguns acreditam que são pouco importantes, ou que têm menor significado do que lhes é atribuído, outros são da opinião que esses acidentes são freqüentes. Verificou-se que só foi relatado diagnóstico fundamentado de dois casos fatais de envenenamento por Bothrops spp em bovinos, e de nenhum por Crotalus spp. Um questionário por nós submetido a patologistas e clínicos veterinários que atuam em diversos Estados do país, revelou apenas raros casos suspeitos de envenenamento ofídico fatal em bovinos no Brasil. Em nossas viagens de estudo e nos trabalhos de diagnóstico nunca estabelecemos o diagnóstico de morte por acidente ofídico em bovinos. Os casos tidos como envenenamento ofídico, na sua grande maioria, são apenas suposições, sem embasamento. Esses "diagnósticos", em geral, são feitos à distância dos animais que morreram, à revelia de exame clínico, necropsia e estudo histopatológico. Importante foi a constatação de que, no Brasil, embora algumas serpentes do gênero Bothrops possam, teoricamente, produzir quantidades suficientes de veneno para matar um bovino adulto, em experimentos realizados, apenas Bothrops alternatus foi capaz de levar a morte um dos três bovinos experimentalmente por ela picados; esse animal tinha apenas 279 kg. Já as serpentes do gênero Crotalus poderiam inocular quantidades letais de veneno para bovinos adultos. Mesmo assim, tanto para Bothrops spp, como para Crotalus spp, há que se considerar que as serpentes, em geral, só inoculam parte do veneno disponível. Esse estudo indica que é necessário melhor investigar as mortes suspeitas de terem sido causadas por acidente ofídico em bovinos no Brasil. O estabelecimento do diagnóstico de morte por envenenamento ofídico, porém, só é poss

  2. 松墨天牛病原菌及其致病性研究%Investigation of Pathogens of Monochamus alternatus in East China and Virulence

    Institute of Scientific and Technical Information of China (English)

    马良进; 张立钦; 林海萍; 毛胜凤

    2009-01-01

    us spp.、木霉Trichoderma spp.和粘质沙雷氏菌Serratia marcescens.经致病性测定表明,球孢白僵菌、金龟子绿僵菌小孢变种、粉拟青霉对松墨天牛有较强的致病力,处理后16d的致死率分别为100%、100%和70%.

  3. Are diseases increasing in the ocean?

    Science.gov (United States)

    Lafferty, Kevin D.; Porter, James W.; Ford, Susan E.

    2004-01-01

    Many factors (climate warming, pollution, harvesting, introduced species) can contribute to disease outbreaks in marine life. Concomitant increases in each of these makes it difficult to attribute recent changes in disease occurrence or severity to any one factor. For example, the increase in disease of Caribbean coral is postulated to be a result of climate change and introduction of terrestrial pathogens. Indirect evidence exists that (a) warming increased disease in turtles; (b) protection, pollution, and terrestrial pathogens increased mammal disease; (c) aquaculture increased disease in mollusks; and (d) release from overfished predators increased sea urchin disease. In contrast, fishing and pollution may have reduced disease in fishes. In other taxa (e.g., sea grasses, crustaceans, sharks), there is little evidence that disease has changed over time. The diversity of patterns suggests there are many ways that environmental change can interact with disease in the ocean.

  4. Taurine increases hippocampal neurogenesis in aging mice

    Directory of Open Access Journals (Sweden)

    Elias Gebara

    2015-05-01

    Full Text Available Aging is associated with increased inflammation and reduced hippocampal neurogenesis, which may in turn contribute to cognitive impairment. Taurine is a free amino acid found in numerous diets, with anti-inflammatory properties. Although abundant in the young brain, the decrease in taurine concentration with age may underlie reduced neurogenesis. Here, we assessed the effect of taurine on hippocampal neurogenesis in middle-aged mice. We found that taurine increased cell proliferation in the dentate gyrus through the activation of quiescent stem cells, resulting in increased number of stem cells and intermediate neural progenitors. Taurine had a direct effect on stem/progenitor cells proliferation, as observed in vitro, and also reduced activated microglia. Furthermore, taurine increased the survival of newborn neurons, resulting in a net increase in adult neurogenesis. Together, these results show that taurine increases several steps of adult neurogenesis and support a beneficial role of taurine on hippocampal neurogenesis in the context of brain aging.

  5. Pension Age Increase: Pro et Contra

    OpenAIRE

    Maleva, T.; Sinyavskaya, O.

    2010-01-01

    Population ageing and growing deficit of Russian pension system place on the agenda an issue of increasing pension age. The article reviews international practices of regulating pension ages. Authors critically discuss the pros and cons of increasing pension ages in Russia from perspectives of demographic situation and trends, labor market, pension system and public attitudes toward this issue. They prove necessity of the effective pension age increasing in Russia and present their views on h...

  6. CO2 Increase: Questions Beyond Climate Change

    OpenAIRE

    Beckmann, G.; Klopries, B.

    1990-01-01

    The increase of the tropospheric C02 (carbon dioxide) concentration is considered by scientists all over the world to be an alarming signal, as becomes evident from the huge amount of literature on the subject. The so-called greenhouse-gas effect, or, more precisely, its anthropogenic component, is mainly caused by the C02 increase. However, other trace gases have also contributed their share. Far-reaching climatic changes, a temperature increase in the tropospheric air, especially in the po...

  7. Familial skin cancer syndromes: Increased melanoma risk.

    Science.gov (United States)

    Ransohoff, Katherine J; Jaju, Prajakta D; Jaju, Prajaka D; Tang, Jean Y; Carbone, Michele; Leachman, Sancy; Sarin, Kavita Y

    2016-03-01

    Phenotypic traits, such as red hair and freckling, increase melanoma risk by 2- to 3-fold. In addition, approximately 10% of melanomas are caused by inherited germline mutations that increase melanoma risk from 4- to >1000-fold. This review highlights the key genes responsible for inherited melanoma, with an emphasis on when a patient should undergo genetic testing. Many genetic syndromes associated with increased melanoma risk are also associated with an increased risk of other cancers. Identification of these high-risk patients is essential for preventive behavior reinforcement, genetic counseling, and ensuring other required cancer screenings.

  8. Does Twitter Increase Perceived Police Legitimacy?

    NARCIS (Netherlands)

    Grimmelikhuijsen, Stephan G; Meijer, Albert J

    2015-01-01

    Social media use has become increasingly popular among police forces. The literature suggests that social media use can increase perceived police legitimacy by enabling transparency and participation. Employing data from a large and representative survey of Dutch citizens (N = 4,492), this article t

  9. Exergames: Increasing Physical Activity through Effective Instruction

    Science.gov (United States)

    Rudella, Jennifer L.; Butz, Jennifer V.

    2015-01-01

    Due to the growing obesity epidemic in the United States, educators must consider new ways to increase physical activity in an effort to address obesity. There are a variety of ways educators can increase physical activity in the classroom, and exergames--video games that require physical movement in order to play--are a modern-day approach to…

  10. Increasing turnover through time in tropical forests

    OpenAIRE

    Phillips, O. L; Gentry, A.H.

    1994-01-01

    Tree turnover rates were assessed at 40 tropical forest sites. Averaged across inventoried forests, turnover, as measured by tree mortality and recruitment, has increased since the 1950's, with an apparent pantropical acceleration since 1980. Among 22 mature forest sites with two or more inventory periods, forest turnover also increased. The trend in forest dynamics may have profound effects on biological diversity.

  11. Wilson's disease; increased aluminum in liver.

    Science.gov (United States)

    Yasui, M; Yoshimasu, F; Yase, Y; Uebayashi, Y

    1979-01-01

    Interaction of trace metal metabolism was studied in a patient with Wilson's dease. Atomic absorption analysis showed markedly increased urinary excretion of copper and aluminum and an increased aluminum content was found in the biopsied liver by neutron activation analysis. These findings suggest a complicated pathogenetic mechanism involving other metals besides copper in the Wilson's disease.

  12. Worldwide Increasing Incidences of Cutaneous Malignant Melanoma

    Directory of Open Access Journals (Sweden)

    Dianne E. Godar

    2011-01-01

    Full Text Available The incidence of cutaneous malignant melanoma (CMM has been increasing at a steady rate in fair-skinned populations around the world for decades. Scientists are not certain why CMM has been steadily increasing, but strong, intermittent UVB (290–320 nm exposures, especially sunburn episodes, probably initiate, CMM, while UVA (321–400 nm passing through glass windows in offices and cars probably promotes it. The CMM incidence may be increasing at an exponential rate around the world, but it definitely decreases with increasing latitude up to ~50°N where it reverses and increases with the increasing latitude. The inversion in the incidence of CMM may occur because there is more UVA relative to UVB for most of the year at higher latitudes. If windows, allowing UVA to enter our indoor-working environment and cars, are at least partly responsible for the increasing incidence of CMM, then UV filters can be applied to reduce the rate of increase worldwide.

  13. Increased muscle glucose uptake during contractions

    DEFF Research Database (Denmark)

    Ploug, Thorkil; Galbo, H; Richter, E A

    1984-01-01

    We reinvestigated the prevailing concept that muscle contractions only elicit increased muscle glucose uptake in the presence of a so-called "permissive" concentration of insulin (Berger et al., Biochem. J. 146: 231-238, 1975; Vranic and Berger, Diabetes 28: 147-163, 1979). Hindquarters from rats...... in severe ketoacidosis were perfused with a perfusate containing insulin antiserum. After 60 min perfusion, electrical stimulation increased glucose uptake of the contracting muscles fivefold. Also, subsequent contractions increased glucose uptake in hindquarters from nondiabetic rats perfused for 1...... Berger et al., 3-O-methylglucose uptake increased during contractions and glucose uptake was negative at rest and zero during contractions. An increase in muscle transport and uptake of glucose during contractions does not require the presence of insulin. Furthermore, glucose transport in contracting...

  14. Why does human culture increase exponentially?

    Science.gov (United States)

    Enquist, M; Ghirlanda, S; Jarrick, A; Wachtmeister, C-A

    2008-08-01

    Historical records show that culture can increase exponentially in time, e.g., in number of poems, musical works, scientific discoveries. We model how human capacities for creativity and cultural transmission may make such an increase possible, suggesting that: (1) creativity played a major role at the origin of human culture and for its accumulation throughout history, because cultural transmission cannot, on its own, generate exponentially increasing amounts of culture; (2) exponential increase in amount of culture can only occur if creativity is positively influenced by culture. The evolution of cultural transmission is often considered the main genetic bottleneck for the origin of culture, because natural selection cannot favor cultural transmission without any culture to transmit. Our models suggest that an increase in individual creativity may have been the first step toward human culture, because in a population of creative individuals there may be enough non-genetic information to favor the evolution of cultural transmission. PMID:18571686

  15. Are increases in cigarette taxation regressive?

    Science.gov (United States)

    Borren, P; Sutton, M

    1992-12-01

    Using the latest published data from Tobacco Advisory Council surveys, this paper re-evaluates the question of whether or not increases in cigarette taxation are regressive in the United Kingdom. The extended data set shows no evidence of increasing price-elasticity by social class as found in a major previous study. To the contrary, there appears to be no clear pattern in the price responsiveness of smoking behaviour across different social classes. Increases in cigarette taxation, while reducing smoking levels in all groups, fall most heavily on men and women in the lowest social class. Men and women in social class five can expect to pay eight and eleven times more of a tax increase respectively, than their social class one counterparts. Taken as a proportion of relative incomes, the regressive nature of increases in cigarette taxation is even more pronounced.

  16. U.S. spending increase planned

    International Nuclear Information System (INIS)

    This article summarizes a paper by Salomon Brothers which published its 14th annual survey and analysis of 1996 worldwide oil and gas exploration and production expenditures. The 28-page report analyzing the December 1995 survey of 275 oil and gas companies indicates the industry is planning a worldwide E and P spending increase of 7.7% this year, over 1995 levels. This is somewhat lower than the now-expected 9.0% increase in 1995 from 1994; however, it is the largest planned increase from a year-end survey since December 1990. Results indicate the recovery in international spending that began in 1995 will be sustained at a more-modest 8.6%, compared to the 14.4% increase last year. Deferrals from 1994 are believed to have contributed to strong 1995 international spending. US spending, conversely, is expected to increase a robust 11.8% in 1996, after a 4.7% advance in 1995. US majors are pacing this increase with a planned 13.4% increase, compared to 9.3% for independents. Canadian operators project a 6.4% decline, even greater than the 4.4% drop registered in 1995

  17. NSF to receive 10% funding increase

    Science.gov (United States)

    Jones, Richard M.

    Work is almost complete on the fiscal year 1994 funding bill for the National Science Foundation. On October 1, Conference Committee Chairs Louis Stokes (D-Ohio) and Barbara Mikulski (D-Md.) and their colleagues completed work on the final version of HR2491, the VA, HUD, Independent Agencies Appropriations Bill. The House and Senate will soon give their final approval to the conference report for HR2491, after which it will lack only the president's signature. NSF's overall budget increase of 10%, raising total funding to %3,005.3 million, is the highest increase among all departments and agencies. Last year's increase in NSF funding was 6.3%.Funding for research and related activities will increase 7% to $1,986 million, a compromise between the 10% House increase and the 4% Senate figure. Last year's bill cut research funding. Education and human resources funding will increase 17% to $569.6 million. This is $13.5 million above the Clinton administration's request. Last year's increase for education and human resources was 4.7%.

  18. Increased skeletal muscle capillarization enhances insulin sensitivity.

    Science.gov (United States)

    Akerstrom, Thorbjorn; Laub, Lasse; Vedel, Kenneth; Brand, Christian Lehn; Pedersen, Bente Klarlund; Lindqvist, Anna Kaufmann; Wojtaszewski, Jørgen F P; Hellsten, Ylva

    2014-12-15

    Increased skeletal muscle capillarization is associated with improved glucose tolerance and insulin sensitivity. However, a possible causal relationship has not previously been identified. Therefore, we investigated whether increased skeletal muscle capillarization increases insulin sensitivity. Skeletal muscle-specific angiogenesis was induced by adding the α1-adrenergic receptor antagonist prazosin to the drinking water of Sprague-Dawley rats (n = 33), whereas 34 rats served as controls. Insulin sensitivity was measured ≥40 h after termination of the 3-wk prazosin treatment, which ensured that prazosin was cleared from the blood stream. Whole body insulin sensitivity was measured in conscious, unrestrained rats by hyperinsulinemic euglycemic clamp. Tissue-specific insulin sensitivity was assessed by administration of 2-deoxy-[(3)H]glucose during the plateau phase of the clamp. Whole body insulin sensitivity increased by ∼24%, and insulin-stimulated skeletal muscle 2-deoxy-[(3)H]glucose disposal increased by ∼30% concomitant with an ∼20% increase in skeletal muscle capillarization. Adipose tissue insulin sensitivity was not affected by the treatment. Insulin-stimulated muscle glucose uptake was enhanced independent of improvements in skeletal muscle insulin signaling to glucose uptake and glycogen synthesis, suggesting that the improvement in insulin-stimulated muscle glucose uptake could be due to improved diffusion conditions for glucose in the muscle. The prazosin treatment did not affect the rats on any other parameters measured. We conclude that an increase in skeletal muscle capillarization is associated with increased insulin sensitivity. These data point toward the importance of increasing skeletal muscle capillarization for prevention or treatment of type 2 diabetes. PMID:25352432

  19. Increasing trends of herpes zoster in Australia.

    Directory of Open Access Journals (Sweden)

    Raina MacIntyre

    Full Text Available Increasing trends in incidence of herpes zoster (HZ have been reported in Australia and internationally. This may reflect the impact of childhood VZV vaccination programs introduced universally in Australia in late 2005. The objective of this study was to evaluate changes in incidence of HZ and PHN in Australia over time, and associated healthcare resource utilisation.Australian data on general practice (GP encounters for HZ, specific antiviral prescribing data from the pharmaceutical benefits scheme, emergency department presentations from the states of NSW and Victoria and national hospitalisation data for HZ were analysed for time trends using regression models. Two time periods (2000-2006 and 2006-2013 were compared which correspond broadly with the pre- and post- universal VZV vaccination period.All data sources showed increasing rates of HZ with age and over time. The GP database showed a significant annual increase in encounters for HZ of 2.5 per 100,000 between 1998 and 2013, and the rates of prescriptions for HZ increased by 4.2% per year between 2002 and 2012. In the 60+ population HZ incidence was estimated to increase from 11.9 to 15.4 per 1,000 persons using GP data or from 12.8 to 14.2 per 1,000 persons using prescription data (p<0.05, between the two periods. Hospitalisation data did not show the same increasing trend over time, except for the age group ≥80 years. Most emergency visits for HZ were not admitted, and showed significant increases over time.The burden of HZ in Australia is substantial, and continues to increase over time. This increase is seen both pre- and post-universal VZV vaccination in 2005, and is most prominent in the older population. The substantial burden of HZ, along with ageing of the Australian population and the importance of healthy ageing, warrants consideration of HZ vaccination for the elderly.

  20. Tobacco use increases susceptibility to bacterial infection

    Directory of Open Access Journals (Sweden)

    Demuth Donald R

    2008-12-01

    Full Text Available Abstract Active smokers and those exposed to secondhand smoke are at increased risk of bacterial infection. Tobacco smoke exposure increases susceptibility to respiratory tract infections, including tuberculosis, pneumonia and Legionnaires disease; bacterial vaginosis and sexually transmitted diseases, such as chlamydia and gonorrhoea; Helicobacter pylori infection; periodontitis; meningitis; otitis media; and post-surgical and nosocomial infections. Tobacco smoke compromises the anti-bacterial function of leukocytes, including neutrophils, monocytes, T cells and B cells, providing a mechanistic explanation for increased infection risk. Further epidemiological, clinical and mechanistic research into this important area is warranted.

  1. Who Benefits from a Minimum Wage Increase?

    OpenAIRE

    John W. Lopresti; Mumford, Kevin J.

    2015-01-01

    This paper addresses the question of how a minimum wage increase affects the wages of low-wage workers. Most studies assume that there is a simple mechanical increase in the wage for workers earning a wage between the old and the new minimum wage, with some studies allowing for spillovers to workers with wages just above this range. Rather than assume that the wages of these workers would have remained constant, this paper estimates how a minimum wage increase impacts a low-wage worker's wage...

  2. Increased transvascular lipoprotein transport in diabetes

    DEFF Research Database (Denmark)

    Jensen, Jan Skov; Feldt-Rasmussen, Bo; Borch-Johnsen, Knut;

    2005-01-01

    likely were not caused by altered hepatic LDL receptor expression, glycosylation of LDL, small LDL size, or medicine use. CONCLUSIONS: Transvascular LDL transport is increased in patients with diabetes mellitus, especially if systolic hypertension or albuminuria is present. Accordingly, lipoprotein flux......CONTEXT: Diabetes is associated with a highly increased risk of atherosclerosis, especially if hypertension or albuminuria is present. OBJECTIVE: We hypothesized that the increased transvascular lipoprotein transport in diabetes may be further accelerated if hypertension or albuminuria is present......, possibly explaining increased intimal lipoprotein accumulation and thus atherosclerosis. DESIGN: The study was cross-sectional and was performed in 1999-2002. SETTING: The study took place in the referral center. PATIENTS: The patients included 60 with diabetes mellitus (27 with type 1 diabetes and 33...

  3. Increasing incidence of diabetes after gestational diabetes

    DEFF Research Database (Denmark)

    Lauenborg, Jeannet; Hansen, Torben; Jensen, Dorte Møller;

    2004-01-01

    To study the incidence of diabetes among women with previous diet-treated gestational diabetes mellitus (GDM) in the light of the general increasing incidence of overweight and diabetes and to identify risk factors for the development of diabetes....

  4. The ways of SOFC systems efficiency increasing

    Energy Technology Data Exchange (ETDEWEB)

    Demin, A.K.; Timofeyeva, N.

    1996-04-01

    The efficiency of solid oxide fuel cells (SOFCs) is described. This paper considers methods to lift the fuel utilization and/or the average cell voltage with the goal of increasing the cell efficiency by improved cell designs.

  5. Autism overflows: increasing prevalence and proliferating theories.

    Science.gov (United States)

    Waterhouse, Lynn

    2008-12-01

    This selective review examines the lack of an explanation for the sharply increasing prevalence of autism, and the lack of any synthesis of the proliferating theories of autism. The most controversial and most widely disseminated notion for increasing prevalence is the measles-mumps-rubella/thimerosal vaccine theory. Less controversial causes that have been proposed include changes in autism diagnostic criteria, increasing services for autism, and growing awareness of the disorder. Regardless of its causes, the increasing prevalence of autism has put pressure on the field of autism research to generate productive and predictive theories of autism. However, the heterogeneity of brain deficits, impaired behaviors, and genetic variants in autism have challenged researchers and theorists, and despite 45 years of research, no standard causal synthesis has emerged. Research going forward should assume that autism is an aggregation of myriad independent disorders of impaired sociality, social cognition, communication, and motor and cognitive skills.

  6. Increase in the Array Television Camera Sensitivity

    Science.gov (United States)

    Shakhrukhanov, O. S.

    A simple adder circuit for successive television frames that enables to considerably increase the sensitivity of such radiation detectors is suggested by the example of array television camera QN902K.

  7. Acute hypoxia increases the cerebral metabolic rate

    DEFF Research Database (Denmark)

    Vestergaard, Mark Bitsch; Lindberg, Ulrich; Aachmann-Andersen, Niels Jacob;

    2016-01-01

    imaging techniques were used to measure global cerebral blood flow and the venous oxygen saturation in the sagittal sinus. Global cerebral metabolic rate of oxygen was quantified from cerebral blood flow and arteriovenous oxygen saturation difference. Concentrations of lactate, glutamate, N......-acetylaspartate, creatine and phosphocreatine were measured in the visual cortex by magnetic resonance spectroscopy. Twenty-three young healthy males were scanned for 60 min during normoxia, followed by 40 min of breathing hypoxic air. Inhalation of hypoxic air resulted in an increase in cerebral blood flow of 15.5% (p = 0.......058), and an increase in cerebral metabolic rate of oxygen of 8.5% (p = 0.035). Cerebral lactate concentration increased by 180.3% ([Formula: see text]), glutamate increased by 4.7% ([Formula: see text]) and creatine and phosphocreatine decreased by 15.2% (p[Formula: see text]). The N-acetylaspartate concentration...

  8. Increased aldosterone: mechanism of hypertension in obesity.

    Science.gov (United States)

    Flynn, Colleen

    2014-05-01

    The prevalence of both obesity and hypertension are increasing worldwide. Hypertension is a common consequence of obesity. Increased central adiposity is associated with increased aldosterone levels and blood pressure in human beings. A number of small studies have shown an association between obesity-mediated hypertension and mechanisms directly linked to increased levels of aldosterone. These studies have shown a trend toward relatively greater blood pressure reduction using aldosterone-receptor blockers compared with other classes of antihypertensive agents. Other than treatment for weight loss, treatment of hypertension with specific antihypertensive medications that block or reduce aldosterone action are appropriate in obese patients. Further research is needed to understand the exact role of the adipocyte in obesity-mediated hypertension.

  9. Ethanologenic bacteria with increased resistance to furfural

    Energy Technology Data Exchange (ETDEWEB)

    Miller, Elliot Norman; Jarboe, Laura R.; Yomano, Lorraine P.; York, Sean W.; Shanmugam, Keelnatham; Ingram, Lonnie O' Neal

    2015-10-06

    The invention relates to bacterium that have increased resistance to furfural and methods of preparation. The invention also relates to methods of producing ethanol using the bacterium and corresponding kits.

  10. Physical activity increases bone mass during growth

    OpenAIRE

    Karlsson, Magnus K; Nordvist, Anders; Karlsson, Caroline

    2008-01-01

    Background: The incidence of fragility fractures has increased during the last half of the 1900?s. One important determinant of fractures is the bone mineral content (BMC) or bone mineral density (BMD), the amount of mineralised bone. If we could increase peak bone mass (the highest value of BMC reached during life) and/or decrease the age-related bone loss, we could possibly improve the skeletal resistance to fracture. Objective: This review evaluates the importance of exercise as a strategy...

  11. Increasing Engagement with the Library via Gamification

    OpenAIRE

    Barr, Matthew; Munro, Kay; Hopfgartner, Frank

    2016-01-01

    One of the main challenges faced by providers of interactive information access systems is to engage users in the use their systems. The library sector in particular can benefit significantly from increased user engagement. In this short paper, we present a preliminary analysis of a university library system that aims to trigger users' extrinsic motivation to increase their interaction with the system. Results suggest that different user groups react in different ways to such 'gamified' systems.

  12. Does Movie Violence Increase Violent Crime?

    OpenAIRE

    Gordon Dahl; Stefano DellaVigna

    2007-01-01

    Laboratory experiments in psychology find that media violence increases aggression in the short run. We analyze whether media violence affects violent crime in the field. We exploit variation in the violence of blockbuster movies from 1995 to 2004, and study the effect on same-day assaults. We find that violent crime decreases on days with larger theater audiences for violent movies. The effect is partly due to voluntary incapacitation: between 6PM and 12AM, a one million increase in the audi...

  13. Leptin increasing sympathetic nerve outflow in obesity

    OpenAIRE

    Simonds, Stephanie E.; Cowley, Michael A.; Enriori, Pablo J.

    2012-01-01

    Obesity is a global problem and effective drug therapy treatment is still unavailable. Obesity develops due to an imbalance between energy intake and energy expenditure (EE). Understanding what happens to EE in obesity may be the key to developing new treatments for obesity. If EE in obesity can be elevated, it could be a potential therapeutic target. We recently discovered that in baseline conditions obese mice have increased EE, in terms of thermogenesis. However, this increase in EE is not...

  14. Population effects of increased climate variation

    OpenAIRE

    Drake, John M.

    2005-01-01

    Global circulation models predict and numerous observations confirm that anthropogenic climate change has altered high-frequency climate variability. However, it is not yet well understood how changing patterns of environmental variation will affect wildlife population dynamics and other ecological processes. Theory predicts that a population's long-run growth rate is diminished and the chance of population extinction is increased as environmental variation increases. This results from the fa...

  15. DUAL RANDOM MODEL OF INCREASING ANNUITY

    Institute of Scientific and Technical Information of China (English)

    HeWenjiong; ZhangYi

    2001-01-01

    The dual random models about the life insurance and social pension insurance have received considerable attention in the recent articles on actuarial theory and applications. This paper discusses a general kind of increasing annuity based on its force of interest accumulationfunction as a general random process. The dual random model of the present value of the benefits of the increasing annuity has been set, and their moments have been calculated under certainconditions.

  16. Minimum wage increases under straightened circumstances

    OpenAIRE

    Addison, John T.; Blackburn, McKinley L.; Cotti, Chad

    2011-01-01

    Do apparently large minimum wage increases in an environment of recession produce clearer evidence of disemployment effects than is typically observed in the new minimum wage literature? This paper augments the sparse literature on the most recent increases in the U.S. minimum wage, using three different data sets and the two main estimation strategies for handling geographically-disparate trends. The evidence is generally unsupportive of negative employment effects, still less of a 'recessio...

  17. Taurine increases hippocampal neurogenesis in aging mice

    OpenAIRE

    Elias Gebara; Florian Udry; Sébastien Sultan; Nicolas Toni

    2015-01-01

    Aging is associated with increased inflammation and reduced hippocampal neurogenesis, which may in turn contribute to cognitive impairment. Taurine is a free amino acid found in numerous diets, with anti-inflammatory properties. Although abundant in the young brain, the decrease in taurine concentration with age may underlie reduced neurogenesis. Here, we assessed the effect of taurine on hippocampal neurogenesis in middle-aged mice. We found that taurine increased cell proliferation in the d...

  18. Increasing incidence of diabetes after gestational diabetes

    DEFF Research Database (Denmark)

    Lauenborg, Jeannet; Hansen, Torben; Jensen, Dorte Møller;

    2004-01-01

    To study the incidence of diabetes among women with previous diet-treated gestational diabetes mellitus (GDM) in the light of the general increasing incidence of overweight and diabetes and to identify risk factors for the development of diabetes.......To study the incidence of diabetes among women with previous diet-treated gestational diabetes mellitus (GDM) in the light of the general increasing incidence of overweight and diabetes and to identify risk factors for the development of diabetes....

  19. Heterostructure optimization for increasing LED efficiency

    Science.gov (United States)

    Rabinovich, Oleg; Legotin, Sergey; Didenko, Sergey; Yakimov, Evgeniy; Osipov, Yuriy; Fedorchenko, Irina

    2016-05-01

    Computer simulations were performed to increase the quantum efficiency of LED by optimizing the nanoheterostructure (NH). Furthermore, the InGaN and AlGaP NHs for LEDs were optimized. On the basis of the optimum NH, ways to further increase the efficiency and the influence of impurities and indium atoms doped into barriers between quantum wells were investigated. The optimum impurity and indium atom concentrations to achieve higher flux were determined.

  20. The Effect of Increasing Mass upon Locomotion

    Science.gov (United States)

    DeWitt, John; Hagan, Donald

    2007-01-01

    The purpose of this investigation was to determine if increasing body mass while maintaining bodyweight would affect ground reaction forces and joint kinetics during walking and running. It was hypothesized that performing gait with increased mass while maintaining body weight would result in greater ground reaction forces, and would affect the net joint torques and work at the ankle, knee and hip when compared to gait with normal mass and bodyweight. Vertical ground reaction force was measured for ten subjects (5M/5F) during walking (1.34 m/s) and running (3.13 m/s) on a treadmill. Subjects completed one minute of locomotion at normal mass and bodyweight and at four added mass (AM) conditions (10%, 20%, 30% and 40% of body mass) in random order. Three-dimensional joint position data were collected via videography. Walking and running were analyzed separately. The addition of mass resulted in several effects. Peak impact forces and loading rates increased during walking, but decreased during running. Peak propulsive forces decreased during walking and did not change during running. Stride time increased and hip extensor angular impulse and positive work increased as mass was added for both styles of locomotion. Work increased at a greater rate during running than walking. The adaptations to additional mass that occur during walking are different than during running. Increasing mass during exercise in microgravity may be beneficial to increasing ground reaction forces during walking and strengthening hip musculature during both walking and running. Future study in true microgravity is required to determine if the adaptations found would be similar in a weightless environment.

  1. Social Closeness Increases Salivary Progesterone in Humans

    OpenAIRE

    Brown, Stephanie L.; Fredrickson, Barbara L; Wirth, Michelle M.; Poulin, Michael J.; Meier, Elizabeth A.; Heaphy, Emily D.; Michael D. Cohen; Schultheiss, Oliver C.

    2009-01-01

    We examined whether interpersonal closeness increases salivary progesterone. One hundred and sixty female college students (80 dyads) were randomly assigned to participate in either a closeness task with a partner versus a neutral task with a partner. Those exposed to the closeness induction had higher levels of progesterone relative to those exposed to the neutral task. Across conditions, progesterone increase one week later predicted the willingness to sacrifice for the partner. These resul...

  2. An experimental increase in stuttering frequency.

    Science.gov (United States)

    Martin, R R; Haroldson, S K

    1988-06-01

    The purpose of the present experiment was to investigate whether stuttering frequency could be increased from baseline by an experimental condition. Initially, 10 adult stutterers spoke spontaneously while alone. Next, they continued speaking, but with an adult, male conversationalist. Finally, the subjects again spoke while alone. Percent stuttering increased in the conversational situation compared with the baseline (alone) situation, and decreased in the final baseline (alone) situation compared with the conversational situation.

  3. Wind increases leaf water use efficiency.

    Science.gov (United States)

    Schymanski, Stanislaus J; Or, Dani

    2016-07-01

    A widespread perception is that, with increasing wind speed, transpiration from plant leaves increases. However, evidence suggests that increasing wind speed enhances carbon dioxide (CO2 ) uptake while reducing transpiration because of more efficient convective cooling (under high solar radiation loads). We provide theoretical and experimental evidence that leaf water use efficiency (WUE, carbon uptake per water transpired) commonly increases with increasing wind speed, thus improving plants' ability to conserve water during photosynthesis. Our leaf-scale analysis suggests that the observed global decrease in near-surface wind speeds could have reduced WUE at a magnitude similar to the increase in WUE attributed to global rise in atmospheric CO2 concentrations. However, there is indication that the effect of long-term trends in wind speed on leaf gas exchange may be compensated for by the concurrent reduction in mean leaf sizes. These unintuitive feedbacks between wind, leaf size and water use efficiency call for re-evaluation of the role of wind in plant water relations and potential re-interpretation of temporal and geographic trends in leaf sizes. PMID:26714739

  4. Energy in China: Coping with increasing demand

    International Nuclear Information System (INIS)

    Sustaining the increasing energy consumption is crucial to future economic growth in China. This report focuses on the current and future situation of energy production and consumption in China and how China is coping with its increasing domestic energy demand. Today, coal is the most important energy resource, followed by oil and hydropower. Most energy resources are located in the inland, whereas the main demand for energy is in the coastal areas, which makes transportation and transmission of energy vital. The industrial sector is the main driver of the energy consumption in China, but the transport sector and the residential sector will increase their share of consumption in China, but the transport sector and the residential sector will increase their share of consumption by 2020. China's energy intensity decreased during the 1990s, but it is still high in a global comparison. China is projected to increase its energy consumption at least two times between 2000 and 2025. The government has an equal focus on energy conservation and to develop the current energy resources. Coal will continue to be the most important fuel, but the demand for oil, hydropower, natural gas and nuclear power will also increase. The main future challenges are transportation of energy resources within China and securing oil supply, both domestic and imports

  5. Wind increases leaf water use efficiency.

    Science.gov (United States)

    Schymanski, Stanislaus J; Or, Dani

    2016-07-01

    A widespread perception is that, with increasing wind speed, transpiration from plant leaves increases. However, evidence suggests that increasing wind speed enhances carbon dioxide (CO2 ) uptake while reducing transpiration because of more efficient convective cooling (under high solar radiation loads). We provide theoretical and experimental evidence that leaf water use efficiency (WUE, carbon uptake per water transpired) commonly increases with increasing wind speed, thus improving plants' ability to conserve water during photosynthesis. Our leaf-scale analysis suggests that the observed global decrease in near-surface wind speeds could have reduced WUE at a magnitude similar to the increase in WUE attributed to global rise in atmospheric CO2 concentrations. However, there is indication that the effect of long-term trends in wind speed on leaf gas exchange may be compensated for by the concurrent reduction in mean leaf sizes. These unintuitive feedbacks between wind, leaf size and water use efficiency call for re-evaluation of the role of wind in plant water relations and potential re-interpretation of temporal and geographic trends in leaf sizes.

  6. Birch Stands Growth Increase in Western Siberia

    Science.gov (United States)

    Kharuk, Viacheslav I.; Kuzmichev, Valeriy V.; Im, Sergey T.; Ranson, Kenneth J.

    2014-01-01

    Birch (Betula pendula Roth) growth within the Western Siberia forest-steppe was analyzed based on long-term (1897-2006) inventory data (height, diameter at breast height [dbh], and stand volume). Analysis of biometry parameters showed increased growth at the beginning of twenty-first century compared to similar stands (stands age = 40-60 years) at the end of nineteenth century. Mean height, dbh, and stem volume increased from 14 to 20 m, from 16 to 22 cm, and from approx. 63 to approx. 220 cu m/ha, respectively. Significant correlations were found between the stands mean height, dbh, and volume on the one hand, and vegetation period length (r(sub s) = 0.71 to 0.74), atmospheric CO2 concentration (r(sub s) = 0.71 to 0.76), and drought index (Standardized Precipitation-Evapotranspiration Index, r(sub s) = -0.33 to -0.51) on the other hand. The results obtained have revealed apparent climate-induced impacts (e.g. increase of vegetation period length and birch habitat drying due to drought increase) on the stands growth. Along with this, a high correlation of birch biometric parameters and [CO2] in ambient air indicated an effect of CO2 fertilization. Meanwhile, further drought increase may switch birch stand growth into decline and greater mortality as has already been observed within the Trans-Baikal forest-steppe ecotone.

  7. Exploring Increased Productivity Through Employee Engagement

    Science.gov (United States)

    Richards, Wayne K., Jr.

    Disengaged employees cost U.S. companies billions of dollars annually in lowered productivity, a cost which has been compounded by the difficult economic situations in the country. The potential for increasing productivity through increased employee engagement was examined in this study. Using personal engagement theory and the theory of planned behavior, the purpose of this phenomenological study was to explore how the experiences of salaried aerospace employees affected productivity and the financial performance of an organization. Interviews were conducted with a purposive sample of 20 aerospace employees whose responses were codified and analyzed to identify themes. The analysis indicated that (a) the lived experiences of employees influenced employee engagement, (b) employee engagement affects organizational commitment and performance, and (c) trust and respect and leadership are essential components to keep employees engaged. Eighty percent of the participants indicated that as employee engagement increases so too does organizational performance. The implications for positive social change include new insights for leaders seeking to increase productivity and financial performance, and to support employee engagement for maintaining sustainability, retaining talent, increasing profits, and improving the economy.

  8. Randomness in Sequence Evolution Increases over Time.

    Science.gov (United States)

    Wang, Guangyu; Sun, Shixiang; Zhang, Zhang

    2016-01-01

    The second law of thermodynamics states that entropy, as a measure of randomness in a system, increases over time. Although studies have investigated biological sequence randomness from different aspects, it remains unknown whether sequence randomness changes over time and whether this change consists with the second law of thermodynamics. To capture the dynamics of randomness in molecular sequence evolution, here we detect sequence randomness based on a collection of eight statistical random tests and investigate the randomness variation of coding sequences with an application to Escherichia coli. Given that core/essential genes are more ancient than specific/non-essential genes, our results clearly show that core/essential genes are more random than specific/non-essential genes and accordingly indicate that sequence randomness indeed increases over time, consistent well with the second law of thermodynamics. We further find that an increase in sequence randomness leads to increasing randomness of GC content and longer sequence length. Taken together, our study presents an important finding, for the first time, that sequence randomness increases over time, which may provide profound insights for unveiling the underlying mechanisms of molecular sequence evolution. PMID:27224236

  9. Passive thermal performance increase in cisterns

    Directory of Open Access Journals (Sweden)

    Najmeh Najafi, Arash Alipour, Seyed Mohamad Ali Najafi

    2014-01-01

    Full Text Available Cisterns are ancient building having two main tasks. First task is storing water in raining seasons for using in dry seasons and the second task is decreasing water temperature, which happens because of the air flow above water surface and evaporating water. It can be stated that by increasing fluid flow above the surface, evaporating increase so increasing in heat transfer happens and decreasing of water temperature is the result. This paper has investigated fluid flow around and inside cisterns with FLUENT software results. Increasing in air flow demand to the cistern is the primary purpose of this study, by changing main parameters of cistern geometry like increasing or decreasing of doom hale diameter, inlet and outlet of wind catcher, elevation of wind catcher and also wind speed. This study introduces the best geometry for cisterns according to maximum air flow demand and minimum volume. Considering that, there is a direct relation between heat transfer and airflow rate in cisterns, so in this study, the influence of mentioned parameters are investigated to reach optimum design for the best performance of cisterns.

  10. Are rates of pediatric bipolar disorder increasing?

    DEFF Research Database (Denmark)

    Kessing, Lars Vedel; Vradi, Eleni; Andersen, Per Kragh

    2014-01-01

    Studies from the USA suggest that rates of pediatric bipolar disorder have increased since the mid-90s, but no study outside the USA has been published on the rates of pediatric bipolar disorder. Further, it is unclear whether an increase in rates reflects a true increase in the illness or more...... diagnostic attention. Using nationwide registers of all inpatients and outpatients contacts to all psychiatric hospitals in Denmark, we investigated (1) gender-specific rates of incident pediatric mania/bipolar disorder during a period from 1995 to 2012, (2) whether age and other characteristics...... for pediatric mania/bipolar disorder changed during the calendar period (1995 to 2003 versus 2004 to 2012), and (3) whether the diagnosis is more often made at first psychiatric contact in recent time compared to earlier according to gender. Totally, 346 patients got a main diagnosis of a manic episode (F30...

  11. Barriers to increasing market-oriented activity

    DEFF Research Database (Denmark)

    Bisp, Søren

    innovation rather than supplying large volumes of consistent quality products. One recommendation from policy-makers and academia to industry is to increase its market-oriented activity. A major reason for the recommendation is that market-oriented activity is considered a major contributor to successful...... product development. Despite the increasing evidence of a positive relationship between above average business performance and a high level of market-oriented activity, normative recommendations on how to increase the level of market-oriented activity is sparse. The scientific contribution of the present......Introduction: The Danish food processing industry faces a situation in which intensified competition in its primary markets and product categories forces several companies to rethink their relative focus in terms of marketing rather than production, or, in other words, in terms of value adding...

  12. Increased rheumatoid factor and deep venous thrombosis

    DEFF Research Database (Denmark)

    Meyer-Olesen, Christine L; Nielsen, Sune F; Nordestgaard, Børge G

    2015-01-01

    BACKGROUND: The risk of deep venous thrombosis is increased in patients with rheumatoid arthritis. We tested the hypothesis that increased concentrations of rheumatoid factor are associated with increased risk of deep venous thrombosis in individuals without autoimmune rheumatic disease...... in the general population. METHODS: We included 54628 participants from the Copenhagen City Heart Study (1981-83) and the Copenhagen General Population Study (2004-12), all with a measured concentration of IgM rheumatoid factor and without autoimmune rheumatic disease or venous thromboembolism. The main outcome...... was incident deep venous thrombosis. There were no losses to follow-up. RESULTS: During 368381 person-years, 670 individuals developed deep venous thrombosis. A rheumatoid factor concentration ≥ vs

  13. Poor Semen Quality Predicts Increased Mortality

    DEFF Research Database (Denmark)

    Jensen, Tina Kold; Bostofte, Erik; Jacobsen, Rune

    Objective: Over recent decades a possible decrease in semen quality and an increase in the incidence of testicular cancer have been reported. In addition, men with poor semen quality have been reported to be at increased risk of developing testicular cancer whereas the risk of other cancers...... is not increased. The long-term survival of men with poor semen quality is, however, unknown. We therefore studied the associations between semen characteristics and subsequent mortality. Back to Top Material and Methods: The Copenhagen Sperm Analysis Laboratory is one of several public semen analysis laboratories...... occurred first. Standardized mortality ratios (SMR) compared with total population of Danish men were calculated according to sperm concentration, motility, and morphology. Back to Top Results: Men with a sperm concentration between 1 and 9 and 10 and 19 million/mL had SMRs of 1.57 (95% CI 1.35–1.81) and 1...

  14. Increased health care utilisation in international adoptees

    DEFF Research Database (Denmark)

    Graff, Heidi Jeannet; Siersma, Volkert Dirk; Kragstrup, Jakob;

    2015-01-01

    after adoption. Our study aimed to theassess health-care utilisation of international adoptees inprimary and secondary care for somatic and psychiatricdiagnoses in a late post-adoption period. Is there an increaseduse of the health-care system in this period, evenwhen increased morbidity in the group...... of allservices in primary care, while in secondary care only fewareas showed an increased long-term morbidity. Conclusion: International adoptees use medical servicesin primary care at a higher rate than non-adoptees someyears after adoption. Excess use of services in secondarycare is also present, but only......Introduction: Several studies have documented thatinternational adoptees have an increased occurrence ofhealth problems and contacts to the health-care systemafter arriving to their new country of residence. This maybe explained by pre-adoption adversities, especially for theperiod immediately...

  15. Innovative Double Bypass Engine for Increased Performance

    Science.gov (United States)

    Manoharan, Sanjivan

    Engines continue to grow in size to meet the current thrust requirements of the civil aerospace industry. Large engines pose significant transportation problems and require them to be split in order to be shipped. Thus, large amounts of time have been spent in researching methods to increase thrust capabilities while maintaining a reasonable engine size. Unfortunately, much of this research has been focused on increasing the performance and efficiencies of individual components while limited research has been done on innovative engine configurations. This thesis focuses on an innovative engine configuration, the High Double Bypass Engine, aimed at increasing fuel efficiency and thrust while maintaining a competitive fan diameter and engine length. The 1-D analysis was done in Excel and then compared to the results from Numerical Propulsion Simulation System (NPSS) software and were found to be within 4% error. Flow performance characteristics were also determined and validated against their criteria.

  16. Nutrient enrichment increases mortality of mangroves.

    Directory of Open Access Journals (Sweden)

    Catherine E Lovelock

    Full Text Available Nutrient enrichment of the coastal zone places intense pressure on marine communities. Previous studies have shown that growth of intertidal mangrove forests is accelerated with enhanced nutrient availability. However, nutrient enrichment favours growth of shoots relative to roots, thus enhancing growth rates but increasing vulnerability to environmental stresses that adversely affect plant water relations. Two such stresses are high salinity and low humidity, both of which require greater investment in roots to meet the demands for water by the shoots. Here we present data from a global network of sites that documents enhanced mortality of mangroves with experimental nutrient enrichment at sites where high sediment salinity was coincident with low rainfall and low humidity. Thus the benefits of increased mangrove growth in response to coastal eutrophication is offset by the costs of decreased resilience due to mortality during drought, with mortality increasing with soil water salinity along climatic gradients.

  17. Alcoholic Cirrhosis Increases Risk for Autoimmune Diseases

    DEFF Research Database (Denmark)

    Grønbæk, Lisbet; Vilstrup, Hendrik; Deleuran, Bent;

    2015-01-01

    BACKGROUND & AIMS: Alcoholic cirrhosis is associated with hyperactivation and dysregulation of the immune system. In addition to its ability to increase risk for infections, it also may increase the risk for autoimmune diseases. We studied the incidence of autoimmune diseases among patients...... with alcohol-associated cirrhosis vs controls in Denmark. METHODS: We collected data from nationwide health care registries to identify and follow up all citizens of Denmark diagnosed with alcoholic cirrhosis from 1977 through 2010. Each patient was matched with 5 random individuals from the population...... diagnosed with alcoholic cirrhosis, 532 developed an autoimmune disease, yielding an overall increased adjusted incidence rate ratio (aIRR) of 1.36 (95% confidence interval [CI], 1.24-1.50). The strongest associations were with Addison's disease (aIRR, 2.47; 95% CI, 1.04-5.85), inflammatory bowel disease (a...

  18. Holographic entropy increases in quadratic curvature gravity

    Science.gov (United States)

    Bhattacharjee, Srijit; Sarkar, Sudipta; Wall, Aron C.

    2015-09-01

    Standard methods for calculating the black hole entropy beyond general relativity are ambiguous when the horizon is nonstationary. We fix these ambiguities in all quadratic curvature gravity theories, by demanding that the entropy be increasing at every time, for linear perturbations to a stationary black hole. Our result matches with the entropy formula found previously in holographic entanglement entropy calculations. We explicitly calculate the entropy increase for Vaidya-like solutions in Ricci-tensor gravity to show that (unlike the Wald entropy) the holographic entropy obeys a second law.

  19. Increasing dominance of IT in ICT convergence

    DEFF Research Database (Denmark)

    Henten, Anders; Tadayoni, Reza

    The aim of the paper is to examine the increasing dominance of IT companies in the converging ICT industry and, on the basis of this development, to contribute to extending the theoretical understanding of market and industry convergence in the ICT area.......The aim of the paper is to examine the increasing dominance of IT companies in the converging ICT industry and, on the basis of this development, to contribute to extending the theoretical understanding of market and industry convergence in the ICT area....

  20. Coal exporters win significant price increases

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2001-04-01

    Coal exports won a 3 dollars/t or almost 8% rise in hard coking coal prices to lift the benchmark Goonyella brand to 42.75 dollars/t. This was followed by major price increases for semi-hard coking coal with the prices of some grades increased by 8 dollars/t or by more than 23%. The new prices will come into effect for the year beginning April, 2001 and acre then first price rises in two years. The article reviews the major Asia countries with their coal supply information. 1 tab., 4 photos.

  1. Role of biofertilizers in increasing tea productivity

    Institute of Scientific and Technical Information of China (English)

    P Nepolean; R Jayanthi; R Vidhya Pallavi; A Balamurugan; T Kuberan; T Beulah; R Premkumar

    2012-01-01

    Objective: To increase the tea productivity using biofertilizers like VAM fungi, phosphobacteria, Azosprillium. Methods: Nitrogen fixing Azospirillum, phosphate solubilising bacteria and VAM fungi were isolated from tea soils of southern India and region specific potential strains were selected based on laboratory, greenhouse, as well as field experiments. A technology for their mass multiplication has been developed. Results: Biofertilizers were showed higher yield at the rate of 5g/seedling and VAM fungi with 15 g of formulation/bush. Conclusions: Multifunctional biofertilizers were used to reduce the chemical fertilizers, diseases and to increase soil fertility in tea.

  2. Fresh inflation with increasing cosmological parameter

    CERN Document Server

    Bellini, M

    2003-01-01

    I study a fresh inflationary model with an increasing F-cosmological parameter. The model provides sufficiently e-folds to solve the flatness/horizon problem and the density fluctuations agree with experimental values. The temperature increases during fresh inflation and reach its maximum value when inflation ends. I find that entropy perturbations always remain below $10^{-4}$ during fresh inflation and become negligible when fresh inflation ends. Hence, the adiabatic fluctuations dominate the primordial spectrum at the end of fresh inflation.

  3. STRATEGIES FOR INCREASING PRODUCTIVITY IN PRODUCTION SYSTEMS

    Directory of Open Access Journals (Sweden)

    Diego Pacheco

    2014-05-01

    Full Text Available The main objective of this article is to point a set of practical strategies that can be adopted to increase the capacity of constraints resources on production systems, when the constraint is inside the factory and not is in the market. To serve this purpose will be presented strategies based on best practices of the Theory of Constraints, Lean Manufacturing and Total Productive Maintenance. This article also presents the mains tools for the deployment of these methodologies. The survey results have provided an objective set of practical strategy that can be used to increase the capacity and productivity of production systems according to the needs of each manufacturing system.

  4. Price of Vitamin B5 Increases Rapidly

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    @@ The export price of vitamin B5 in China has increased constantly in 2007. According to the analysis made by experts, the demand of vitamin B5 has not grown greatly. The main reason for the price rise is the change of the supply mode.

  5. Explaining Increases in Higher Education Costs

    OpenAIRE

    Robert B Archibald; David H. Feldman

    2006-01-01

    This paper presents new evidence on the conflict between two competing explanations of the increase in college costs, the cost disease theory of William Baumol and William Bowen and the revenue theory of cost of Howard Bowen. Using cross section data, the paper demonstrates that the cost disease explanation dominates.

  6. Voluntary attention increases perceived spatial frequency

    OpenAIRE

    Abrams, Jared; Barbot, Antoine; Carrasco, Marisa

    2010-01-01

    Voluntary covert attention selects relevant sensory information for prioritized processing. The behavioral and neural consequences of such selection have been extensively documented, but its phenomenology has received little empirical investigation. Involuntary attention increases perceived spatial frequency (Gobell & Carrasco, 2005), but involuntary attention can differ from voluntary attention in its effects on performance in tasks mediated by spatial resolution (Yeshurun, Montagna, & Carra...

  7. Increasing Student Participation in Science Fair Competitions

    Science.gov (United States)

    Miles, Rhea

    2012-01-01

    In the summer of 2009, 22 African American middle school students in eastern North Carolina became participants in the Reach Up program to increase the number of underrepresented students participating in science-, technology-, engineering-, and mathematics-related activities. One of the goals of the program was for these students to participate…

  8. Increasing Efficiency: A Process-Oriented Approach.

    Science.gov (United States)

    Harbour, Jerry L.

    1993-01-01

    Discussion of the need to increase efficiency focuses on a process-oriented approach for systematically identifying and minimizing non-value-adding process steps to analyze and improve tasks, services, and production. Highlights include a historical perspective, a discussion of wasted efforts, and a case study. (Contains 16 references.) (LRW)

  9. Fullerene derivatives with increased dielectric constants

    NARCIS (Netherlands)

    Jahani, Fatemeh; Torabi, Solmaz; Chiechi, Ryan C; Koster, L Jan Anton; Hummelen, Jan C

    2014-01-01

    The invention of new organic materials with high dielectric constants is of extreme importance for the development of organic-based devices such as organic solar cells. We report on a synthetic way to increase the dielectric constant of fullerene derivatives. It is demonstrated that introducing trie

  10. Increased Gonorrhea Cases - Utah, 2009-2014.

    Science.gov (United States)

    Watson, Joanna; Carlile, Jerry; Dunn, Angela; Evans, Megan; Fratto, Erin; Hartsell, Joel; Meinor, Lynn; Mietchen, Matthew; Nakashima, Allyn

    2016-01-01

    Gonorrhea (caused by infection with Neisseria gonorrhoeae) is the second most commonly reported notifiable disease in the United States (1). Left untreated, gonorrhea is associated with serious long-term adverse health effects, including pelvic inflammatory disease, ectopic pregnancy, and infertility. Infection also facilitates transmission of human immunodeficiency virus (2,3). Effective gonorrhea control relies upon early detection and effective antimicrobial treatment. To assess gonorrhea rate trends in Utah, the Utah Department of Health (UDOH) analyzed Utah National Electronic Disease Surveillance System (UT-NEDSS) data for the state during 2009-2014. After declining during 2009-2011, the statewide gonorrhea rate increased fivefold to 49 cases per 100,000 population in 2014. During 2009-2014, the proportion of cases among women increased from 21% to 39% (decreasing among males from 79% to 61%). Among male patients, the proportion who identified as men who have sex with men (MSM) decreased from 67% to 42%. These demographic changes suggest that increased heterosexual transmission of gonorrhea in Utah might be occurring. Health departments need to work with providers to ensure populations at high risk are being screened and properly treated for gonorrhea. Clinicians need to be aware of increases in the risk for infection among women and non-MSM males when making screening and testing decisions and educate their patients regarding gonorrhea transmission and prevention practices. PMID:27583786

  11. Voluntary attention increases perceived spatial frequency.

    Science.gov (United States)

    Abrams, Jared; Barbot, Antoine; Carrasco, Marisa

    2010-08-01

    Voluntary covert attention selects relevant sensory information for prioritized processing. The behavioral and neural consequences of such selection have been extensively documented, but its phenomenology has received little empirical investigation. Involuntary attention increases perceived spatial frequency (Gobell & Carrasco, 2005), but involuntary attention can differ from voluntary attention in its effects on performance in tasks mediated by spatial resolution (Yeshurun, Montagna, & Carrasco, 2008). Therefore, we ask whether voluntary attention affects the subjective appearance of spatial frequency--a fundamental dimension of visual perception underlying spatial resolution. We used a demanding rapid serial visual presentation task to direct voluntary attention and measured perceived spatial frequency at the attended and unattended locations. Attention increased the perceived spatial frequency of suprathreshold stimuli and also improved performance on a concurrent orientation discrimination task. In the control experiment, we ruled out response bias as an alternative account by using a lengthened interstimulus interval, which allows observers to disengage attention from the cued location. In contrast to the main experiment, the observers showed neither increased perceived spatial frequency nor improved orientation discrimination at the attended location. Thus, this study establishes that voluntary attention increases perceived spatial frequency. This phenomenological consequence links behavioral and neurophysiological studies on the effects of attention.

  12. Codon optimizing for increased membrane protein production

    DEFF Research Database (Denmark)

    Mirzadeh, K.; Toddo, S.; Nørholm, Morten;

    2016-01-01

    Reengineering a gene with synonymous codons is a popular approach for increasing production levels of recombinant proteins. Here we present a minimalist alternative to this method, which samples synonymous codons only at the second and third positions rather than the entire coding sequence...

  13. NEPAL AND INDIA MAY INCREASE POWER TRADING

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Nepal and India may increase the quantity of power exchanged between the two nations to meet shortages in bordering areas. Technical paneles from the two countries have recommended that the power exchange be raised from 50MW to 150MW, according to Bholanath Chalise, chief of the state-run Nepal Electricity Authority(NEA).

  14. Potential transgenic routes to increase tree biomass.

    Science.gov (United States)

    Dubouzet, Joseph G; Strabala, Timothy J; Wagner, Armin

    2013-11-01

    Biomass is a prime target for genetic engineering in forestry because increased biomass yield will benefit most downstream applications such as timber, fiber, pulp, paper, and bioenergy production. Transgenesis can increase biomass by improving resource acquisition and product utilization and by enhancing competitive ability for solar energy, water, and mineral nutrients. Transgenes that affect juvenility, winter dormancy, and flowering have been shown to influence biomass as well. Transgenic approaches have increased yield potential by mitigating the adverse effects of prevailing stress factors in the environment. Simultaneous introduction of multiple genes for resistance to various stress factors into trees may help forest trees cope with multiple or changing environments. We propose multi-trait engineering for tree crops, simultaneously deploying multiple independent genes to address a set of genetically uncorrelated traits that are important for crop improvement. This strategy increases the probability of unpredictable (synergistic or detrimental) interactions that may substantially affect the overall phenotype and its long-term performance. The very limited ability to predict the physiological processes that may be impacted by such a strategy requires vigilance and care during implementation. Hence, we recommend close monitoring of the resultant transgenic genotypes in multi-year, multi-location field trials. PMID:24094056

  15. Increasing security in inter-chip communication

    Energy Technology Data Exchange (ETDEWEB)

    Edwards, Nathan J; Hamlet, Jason; Bauer, Todd; Helinski, Ryan

    2014-10-28

    An apparatus for increasing security in inter-chip communication includes a sending control module, a communication bus, and a receiving control module. The communication bus is coupled between the sending control module and the receiving control module. The sending control module operates to send data on the communication bus, disable the communication bus when threats are detected, or both.

  16. Hanford Waste Vitrification Plant capacity increase options

    International Nuclear Information System (INIS)

    Studies are being conducted by the Hanford Waste Vitrification Plant (HWVP) Project on ways to increase the waste processing capacity within the current Vitrification Building structural design. The Phase 1 study on remote systems concepts identification and extent of capacity increase was completed. The study concluded that the HWVP capacity could be increased to four times the current capacity with minor design adjustments to the fixed facility design, and the required design changes would not impact the current footprint of the vitrification building. A further increase in production capacity may be achievable but would require some technology development, verification testing, and a more systematic and extensive engineering evaluation. The primary changes included a single advance melter with a higher capacity, new evaporative feed tank, offgas quench collection tank, ejector venturi scrubbers, and additional inner canister closure station,a smear test station, a new close- coupled analytical facility, waste hold capacity of 400,000 gallon, the ability to concentrate out-of-plant HWVP feed to 90 g/L waste oxide concentration, and limited changes to the current base slab construction package

  17. Robotic Bipedal Running: Increasing disturbance rejection

    NARCIS (Netherlands)

    Karssen, J.G.D.

    2013-01-01

    The goal of the research presented in this thesis is to increase the understanding of the human running gait. The understanding of the human running gait is essential for the development of devices, such as prostheses and orthoses, that enable disabled people to run or that enable able people to inc

  18. Increased textural complexity in food enhances satiation.

    Science.gov (United States)

    Larsen, Danaé S; Tang, Jingyuan; Ferguson, Lynnette R; James, Bryony J

    2016-10-01

    For the first time this study has shown a direct effect of food textural complexity on satiation. Independent of oral processing time, increasing the textural complexity of a food significantly decreased food intake. Foods with complex textures stimulate many sensory perceptions during oral processing, with a succession of textures perceived between first bite and swallow. Previously the impact of texture on satiation (commonly tested by increasing viscosities of semi-solids) has been explained by texture's influence on oral processing time; a long oral processing time enhances satiation. The results of the current study show that subjects in a randomised cross-over trial who consumed a "starter" (preload) model food with high textural complexity went on to eat significantly less of a two course ad libitum meal. Subjects who consumed a "starter" model food with low textural complexity, but with the same flavour, energy density and oral processing time, ate significantly more of the same ad libitum meal. The results show that increasing the number of textures perceived during chewing of a solid food triggers the satiation response earlier than when chewing a less texturally complex food. Increasing textural complexity of manufactured foods, to allow for greater sensory stimulation per bite, could potentially be used as a tool to enhance the satiation response and decrease food intake. PMID:27235823

  19. Smart Partnerships to Increase Equity in Education

    Science.gov (United States)

    Leahy, Margaret; Davis, Niki; Lewin, Cathy; Charania, Amina; Nordin, Hasniza; Orlic, Davor; Butler, Deirdre; Lopez-Fernadez, Olatz

    2016-01-01

    This exploratory analysis of smart partnerships identifies the risk of increasing the digital divide with the deployment of data analytics. Smart partnerships in education appear to include a process of evolution into a synergy of strategic and holistic approaches that enhance the quality of education with digital technologies, harnessing ICT…

  20. Using Mobile Learning to Increase Environmental Awareness

    Science.gov (United States)

    Uzunboylu, Huseyin; Cavus, Nadire; Ercag, Erinc

    2009-01-01

    Mobile learning or m-learning, a relatively new concept, has attracted the interest of educators, researchers, and companies developing learning systems and instructional materials. This study investigated the use of integrating use of mobile technologies, data services, and multimedia messaging systems to increase students' use of mobile…

  1. Increasing Incidence of Juvenile Thyrotoxicosis in Denmark

    DEFF Research Database (Denmark)

    Kjær, R. H.; Andersen, M. S.; Hansen, D.

    2015-01-01

    . Additional data were collected on children diagnosed with GD in 2008-2012. Results: In total, 237 patients with juvenile thyrotoxicosis (JT) were identified. The overall IR in 1998-2012 was 1.58/100,000 person-years and has increased significantly from 0.79/100,000 person-years in 1982-1988 (p

  2. Increased cerebral water content in hemodialysis patients.

    Directory of Open Access Journals (Sweden)

    Kathrin Reetz

    Full Text Available Little information is available on the impact of hemodialysis on cerebral water homeostasis and its distribution in chronic kidney disease. We used a neuropsychological test battery, structural magnetic resonance imaging (MRI and a novel technique for quantitative measurement of localized water content using 3T MRI to investigate ten hemodialysis patients (HD on a dialysis-free day and after hemodialysis (2.4±2.2 hours, and a matched healthy control group with the same time interval. Neuropsychological testing revealed mainly attentional and executive cognitive dysfunction in HD. Voxel-based-morphometry showed only marginal alterations in the right inferior medial temporal lobe white matter in HD compared to controls. Marked increases in global brain water content were found in the white matter, specifically in parietal areas, in HD patients compared to controls. Although the global water content in the gray matter did not differ between the two groups, regional increases of brain water content in particular in parieto-temporal gray matter areas were observed in HD patients. No relevant brain hydration changes were revealed before and after hemodialysis. Whereas longer duration of dialysis vintage was associated with increased water content in parieto-temporal-occipital regions, lower intradialytic weight changes were negatively correlated with brain water content in these areas in HD patients. Worse cognitive performance on an attention task correlated with increased hydration in frontal white matter. In conclusion, long-term HD is associated with altered brain tissue water homeostasis mainly in parietal white matter regions, whereas the attentional domain in the cognitive dysfunction profile in HD could be linked to increased frontal white matter water content.

  3. Amphetamine enhances endurance by increasing heat dissipation.

    Science.gov (United States)

    Morozova, Ekaterina; Yoo, Yeonjoo; Behrouzvaziri, Abolhassan; Zaretskaia, Maria; Rusyniak, Daniel; Zaretsky, Dmitry; Molkov, Yaroslav

    2016-09-01

    Athletes use amphetamines to improve their performance through largely unknown mechanisms. Considering that body temperature is one of the major determinants of exhaustion during exercise, we investigated the influence of amphetamine on the thermoregulation. To explore this, we measured core body temperature and oxygen consumption of control and amphetamine-trea ted rats running on a treadmill with an incrementally increasing load (both speed and incline). Experimental results showed that rats treated with amphetamine (2 mg/kg) were able to run significantly longer than control rats. Due to a progressively increasing workload, which was matched by oxygen consumption, the control group exhibited a steady increase in the body temperature. The administration of amphetamine slowed down the temperature rise (thus decreasing core body temperature) in the beginning of the run without affecting oxygen consumption. In contrast, a lower dose of amphetamine (1 mg/kg) had no effect on measured parameters. Using a mathematical model describing temperature dynamics in two compartments (the core and the muscles), we were able to infer what physiological parameters were affected by amphetamine. Modeling revealed that amphetamine administration increases heat dissipation in the core. Furthermore, the model predicted that the muscle temperature at the end of the run in the amphetamine-treated group was significantly higher than in the control group. Therefore, we conclude that amphetamine may mask or delay fatigue by slowing down exercise-induced core body temperature growth by increasing heat dissipation. However, this affects the integrity of thermoregulatory system and may result in potentially dangerous overheating of the muscles.

  4. A mechanistic study to increase understanding of titanium dioxide nanoparticles-increased plasma glucose in mice.

    Science.gov (United States)

    Hu, Hailong; Li, Li; Guo, Qian; Jin, Sanli; Zhou, Ying; Oh, Yuri; Feng, Yujie; Wu, Qiong; Gu, Ning

    2016-09-01

    Titanium dioxide nanoparticle (TiO2 NP) is an authorized food additive. Previous studies determined oral administration of TiO2 NPs increases plasma glucose in mice via inducing insulin resistance. An increase in reactive oxygen species (ROS) has been considered the possible mechanism of increasing plasma glucose. However, persistently high plasma glucose is also a mechanism of increasing ROS. This study aims to explore whether TiO2 NPs increase plasma glucose via ROS. We found after oral administration of TiO2 NPs, an increase in ROS preceded an increase in plasma glucose. Subsequently, mice were treated with two antioxidants (resveratrol and vitamin E) at the same time as oral administration of TiO2 NPs. Results showed resveratrol and vitamin E reduced TiO2 NPs-increased ROS. An increase in plasma glucose was also inhibited. Further research showed resveratrol and vitamin E inhibited the secretion of TNF-α and IL-6, and the phosphorylation of JNK and p38 MAPK, resulting in improved insulin resistance. These results suggest TiO2 NPs increased ROS levels, and then ROS activated inflammatory cytokines and phosphokinases, and thus induced insulin resistance, resulting in an increase in plasma glucose. Resveratrol and vitamin E can reduce TiO2 NPs-increased ROS and thereby inhibit an increase in plasma glucose in mice.

  5. A mechanistic study to increase understanding of titanium dioxide nanoparticles-increased plasma glucose in mice.

    Science.gov (United States)

    Hu, Hailong; Li, Li; Guo, Qian; Jin, Sanli; Zhou, Ying; Oh, Yuri; Feng, Yujie; Wu, Qiong; Gu, Ning

    2016-09-01

    Titanium dioxide nanoparticle (TiO2 NP) is an authorized food additive. Previous studies determined oral administration of TiO2 NPs increases plasma glucose in mice via inducing insulin resistance. An increase in reactive oxygen species (ROS) has been considered the possible mechanism of increasing plasma glucose. However, persistently high plasma glucose is also a mechanism of increasing ROS. This study aims to explore whether TiO2 NPs increase plasma glucose via ROS. We found after oral administration of TiO2 NPs, an increase in ROS preceded an increase in plasma glucose. Subsequently, mice were treated with two antioxidants (resveratrol and vitamin E) at the same time as oral administration of TiO2 NPs. Results showed resveratrol and vitamin E reduced TiO2 NPs-increased ROS. An increase in plasma glucose was also inhibited. Further research showed resveratrol and vitamin E inhibited the secretion of TNF-α and IL-6, and the phosphorylation of JNK and p38 MAPK, resulting in improved insulin resistance. These results suggest TiO2 NPs increased ROS levels, and then ROS activated inflammatory cytokines and phosphokinases, and thus induced insulin resistance, resulting in an increase in plasma glucose. Resveratrol and vitamin E can reduce TiO2 NPs-increased ROS and thereby inhibit an increase in plasma glucose in mice. PMID:27430421

  6. Increased Baseline C-Reactive Protein Concentrations Are Associated with Increased Risk of Infections

    DEFF Research Database (Denmark)

    Zacho, Jeppe; Benfield, Thomas; Tybjærg-Hansen, Anne;

    2016-01-01

    BACKGROUND: The acute-phase reactant C-reactive protein (CRP) increases rapidly during an infection. We tested the hypothesis that chronic low-level increases in CRP are associated with an increased risk of infectious disease. METHODS: We studied 9660 individuals from a prospective general...... combinations examined conferred an increased risk of infectious disease. CONCLUSIONS: Chronic low-level CRP increases were associated with increased risk of bacterial infections, gram-negative infections in particular. Genotypes associated with increases in CRP were not associated with increased risk...... population cohort, including 3592 in whom infectious disease developed, and another 60 896 individuals from a cross-sectional general population study, of whom 13 332 developed infectious disease; 55% were women, and the mean age was 57 years. Hospital diagnoses of infections in 1977-2010 were based...

  7. Increasing pulse wave velocity in a realistic cardiovascular model does not increase pulse pressure with age

    OpenAIRE

    Mohiuddin, Mohammad W.; Rihani, Ryan J.; Laine, Glen A.; Quick, Christopher M.

    2012-01-01

    The mechanism of the well-documented increase in aortic pulse pressure (PP) with age is disputed. Investigators assuming a classical windkessel model believe that increases in PP arise from decreases in total arterial compliance (Ctot) and increases in total peripheral resistance (Rtot) with age. Investigators assuming a more sophisticated pulse transmission model believe PP rises because increases in pulse wave velocity (cph) make the reflected pressure wave arrive earlier, augmenting systol...

  8. Playing violent video games increases intergroup bias.

    Science.gov (United States)

    Greitemeyer, Tobias

    2014-01-01

    Previous research has shown how, why, and for whom violent video game play is related to aggression and aggression-related variables. In contrast, less is known about whether some individuals are more likely than others to be the target of increased aggression after violent video game play. The present research examined the idea that the effects of violent video game play are stronger when the target is a member of an outgroup rather than an ingroup. In fact, a correlational study revealed that violent video game exposure was positively related to ethnocentrism. This relation remained significant when controlling for trait aggression. Providing causal evidence, an experimental study showed that playing a violent video game increased aggressive behavior, and that this effect was more pronounced when the target was an outgroup rather than an ingroup member. Possible mediating mechanisms are discussed.

  9. Has risk associated with smoking increased?

    DEFF Research Database (Denmark)

    Prescott, E; Osler, M; Andersen, P K;

    1997-01-01

    BACKGROUND: Two recent much cited publications have raised the concern that risk associated with cigarette smoking has so far been underestimated. In this study we wish to determine whether excess all-cause mortality associated with smoking has increased during the last 20-30 years in a study...... population representative of the general Danish population and whether any such changes relate to changes in smoking behaviour. METHODS: Pooled data from three prospective population studies conducted in Copenhagen with detailed information on smoking habits. A total of 31,194 subjects, 17,669 males and 13....... RESULTS: Male smokers' exposure did not change during the study period whereas female smokers' exposure to tobacco increased in terms of age at smoking onset, quantity smoked and depth of inhalation. During follow-up 5744 males and 2900 females died. In males, death rate ratios (comparing continuous...

  10. Playing violent video games increases intergroup bias.

    Science.gov (United States)

    Greitemeyer, Tobias

    2014-01-01

    Previous research has shown how, why, and for whom violent video game play is related to aggression and aggression-related variables. In contrast, less is known about whether some individuals are more likely than others to be the target of increased aggression after violent video game play. The present research examined the idea that the effects of violent video game play are stronger when the target is a member of an outgroup rather than an ingroup. In fact, a correlational study revealed that violent video game exposure was positively related to ethnocentrism. This relation remained significant when controlling for trait aggression. Providing causal evidence, an experimental study showed that playing a violent video game increased aggressive behavior, and that this effect was more pronounced when the target was an outgroup rather than an ingroup member. Possible mediating mechanisms are discussed. PMID:24085715

  11. Increasing the efficiency of solar thermal panels

    Science.gov (United States)

    Dobrnjac, M.; Latinović, T.; Dobrnjac, S.; Živković, P.

    2016-08-01

    The popularity of solar heating systems is increasing for several reasons. These systems are reliable, adaptable and pollution-free, because the renewable solar energy is used. There are many variants of solar systems in the market mainly constructed with copper pipes and absorbers with different quality of absorption surface. Taking into account the advantages and disadvantages of existing solutions, in order to increase efficiency and improve the design of solar panel, the innovative solution has been done. This new solar panel presents connection of an attractive design and the use of constructive appropriate materials with special geometric shapes. Hydraulic and thermotechnical tests that have been performed on this panel showed high hydraulic and structural stability. Further development of the solar panel will be done in the future in order to improve some noticed disadvantages.

  12. Can increased organic consumption mitigate climate changes?

    DEFF Research Database (Denmark)

    Heerwagen, Lennart Ravn; Andersen, Laura Mørch; Christensen, Tove;

    2014-01-01

    consumers. As some consumers believe that climate change can be mitigated by consuming organic food, the authors propose that this is taken into account in the development of organic farming. Originality/value – The authors propose a shift from analysing the climate-friendliness of production to addressing......Purpose – The purpose of this paper is to investigate the evidence for a positive correlation between increased consumption of organic products and potential climate change mitigation via decreased consumption of meat and it is discussed to what extent organic consumption is motivated by climate...... correlation between increasing organic budget shares and decreasing meat budget shares is found. People include food-related behaviour such as the purchase of organic food and reduced meat consumption as ways to mitigate climate change. However, other behavioural modifications such as reduction of car usage...

  13. Do fat supplements increase physical performance?

    Science.gov (United States)

    Macaluso, Filippo; Barone, Rosario; Catanese, Patrizia; Carini, Francesco; Rizzuto, Luigi; Farina, Felicia; Di Felice, Valentina

    2013-02-07

    Fish oil and conjugated linoleic acid (CLA) belong to a popular class of food supplements known as "fat supplements", which are claimed to reduce muscle glycogen breakdown, reduce body mass, as well as reduce muscle damage and inflammatory responses. Sport athletes consume fish oil and CLA mainly to increase lean body mass and reduce body fat. Recent evidence indicates that this kind of supplementation may have other side-effects and a new role has been identified in steroidogenensis. Preliminary findings demonstrate that fish oil and CLA may induce a physiological increase in testosterone synthesis. The aim of this review is to describe the effects of fish oil and CLA on physical performance (endurance and resistance exercise), and highlight the new results on the effects on testosterone biosynthesis. In view of these new data, we can hypothesize that fat supplements may improve the anabolic effect of exercise.

  14. INCREASING DIVERSITY IN OUR SCHOOLS OF NURSING.

    Science.gov (United States)

    Neubrander, Judy; Metcalfe, Sharon E

    2016-01-01

    This article will review one school's quest to address the multi-level social, historical, environmental and structural determinants faced by under-represented ethnic minorities (UREM) and disadvantaged background (DB) students as they seek entrance into a nursing program. Nursing Network Careers and Technology (NN-CAT) provides a nursing career network for underrepresented and disadvantaged students in western North Carolina and has increased the number of underrepresented and disadvantaged students who are admitted, retained and graduate with a bachelor's degree in nursing from Western Carolina University. Initial data from this NN-CAT program have demonstrated that addressing social determinants and eliminating barriers can increase the number of UREM and educationally disadvantaged students who successfully matriculate in our schools of Nursing and subsequently graduate. These nurses then enter the workforce and provide culturally meaningful care in their local communities. PMID:27439229

  15. Increase of Organization in Complex Systems

    CERN Document Server

    Georgiev, Georgi Yordanov; Gombos, Erin; Vinod, Amrit; Hoonjan, Gajinder

    2013-01-01

    Measures of complexity and entropy have not converged to a single quantitative description of levels of organization of complex systems. The need for such a measure is increasingly necessary in all disciplines studying complex systems. To address this problem, starting from the most fundamental principle in Physics, here a new measure for quantity of organization and rate of self-organization in complex systems based on the principle of least (stationary) action is applied to a model system - the central processing unit (CPU) of computers. The quantity of organization for several generations of CPUs shows a double exponential rate of change of organization with time. The exact functional dependence has a fine, S-shaped structure, revealing some of the mechanisms of self-organization. The principle of least action helps to explain the mechanism of increase of organization through quantity accumulation and constraint and curvature minimization with an attractor, the least average sum of actions of all elements ...

  16. Increasing the Size of Microwave Popcorn

    Science.gov (United States)

    Smoyer, Justin

    2005-03-01

    Each year Americans consume approximately 17 billion quarts of popcorn. Since the 1940s, microwaves have been the heating source of choice for most. By treating the popcorn mechanism as a thermodynamic system, it has been shown mathematically and experimentally that reducing the surrounding pressure of the unpopped kernels, results in an increased volume of the kernels [Quinn et al, http://xxx.lanl.gov/abs/cond-mat/0409434 v1 2004]. In this project an alternate method of popping with the microwave was used to further test and confirm this hypothesis. Numerous experimental trials where run to test the validity of the theory. The results show that there is a significant increase in the average kernel size as well as a reduction in the number of unpopped kernels.

  17. Minimal Increase Network Coding for Dynamic Networks.

    Science.gov (United States)

    Zhang, Guoyin; Fan, Xu; Wu, Yanxia

    2016-01-01

    Because of the mobility, computing power and changeable topology of dynamic networks, it is difficult for random linear network coding (RLNC) in static networks to satisfy the requirements of dynamic networks. To alleviate this problem, a minimal increase network coding (MINC) algorithm is proposed. By identifying the nonzero elements of an encoding vector, it selects blocks to be encoded on the basis of relationship between the nonzero elements that the controls changes in the degrees of the blocks; then, the encoding time is shortened in a dynamic network. The results of simulations show that, compared with existing encoding algorithms, the MINC algorithm provides reduced computational complexity of encoding and an increased probability of delivery. PMID:26867211

  18. Increased ocean heat transports and warmer climate

    Science.gov (United States)

    Rind, D.; Chandler, M.

    1991-01-01

    The impact of an increased ocean heat transport on climate is investigated in the framework of the GISS GMC model described by Hansen et al. (1983), using two scenarios: one starting from warmer polar temperatures/no sea ice and the other from the current ocean conditions. A 20-percent increase in cross-equatorial heat transport was sufficient to melt all sea ice; it resulted in a climate that was 2 C warmer for the global average, with values some 20-deg warmer at high altitudes and 1-deg warmer near the equator. It is suggested that the hydrological and dynamical changes associated with this different climate regime may be self-sustaining and, as such, would account for the high-latitude warmth of climates in the Mesozoic and Tertiary periods and the decadenal-scale climate fluctuations during the Holocene.

  19. Leptospirosis: a globally increasing zoonotic disease.

    LENUS (Irish Health Repository)

    Rock, Clare

    2010-01-01

    A 27-year-old previously healthy man was admitted to the intensive care unit with severe jaundice, dyspnoea with haemoptysis, anaemia, thrombocytopenia and acute renal injury. He had no recent history of foreign travel but had been building a shed in his back garden in Cork, Ireland, for the preceding week. The patient\\'s history, clinical observations, haematological and radiological results were all consistent with icteric leptospirosis or Weil\\'s disease. This was confirmed on serological testing. He completed 7 days intravenous ceftriaxone and made a complete recovery. While endemic in tropical climates, leptospirosis incidence is increasing in temperate climates. Recent cases seen in temperate climates can be severe, particularly with pulmonary manifestations. The report of this case serves to increase awareness of this re-emerging potentially fatal infectious disease.

  20. Coyote Attacks: An Increasing Suburban Problem

    OpenAIRE

    Timm, Robert M.; Baker, Rex O.; Bennett, Joe R.; Coolahan, Craig C.

    2004-01-01

    Coyote attacks on humans and pets have increased within the past 5 years in California. We discuss documented occurrences of coyote aggression and attacks on people, using data from USDA Wildlife Services, the California Department of Fish & Game, and other sources. Forty-eight such attacks on children and adults were verified from 1998 through 2003, compared to 41 attacks during the period 1988 through 1997; most incidents occurred in Southern California near the suburban-wildland interfac...