23 results on '"Rodrigues-Simioni L"'
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2. INTRASPECIFIC VARIATION IN NEUROTOXIC AND MYOTOXIC ACTIVITIES OF Bothrops neuwiedii VENOMS
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BORJA-OLIVEIRA, C. R., SOARES, A. M., ZAMUNÉR, S. R., HYSLOP, S., GIGLIO, J. R., PRADO-FRANCESCHI, J., and RODRIGUES-SIMIONI, L.
- Subjects
myotoxicity ,neuromuscular blockade ,intraspecific variation ,neurotoxicity ,complex mixtures ,Bothrops neuwiedii venom - Abstract
Snake venoms frequently vary in composition. In this work, we compared the neurotoxic and myotoxic activities of 16 lots of Bothrops neuwiedii venoms from different regions of Brazil, using chick biventer cervicis preparations. The neuromuscular blockade varied from 2% to 100% after 120 min incubation with venoms (50 mug/ml). In all cases, this blockade was irreversible and concentration-dependent; at low concentrations (10-20 mug/ml), 15 of the 16 venom lots failed to abolish responses to acetylcholine (110 muM), but blocked responses to KCl (13.4 muM), and induced contracture. At 5-20 mug/ml, the most active venom totally blocked twitch-tension without affecting responses to acetylcholine and KCl. Polyacrylamide gel electrophoresis for basic proteins showed that the most active samples contained a band that was absent in the less active venoms. These results indicate that there may be considerable intraspecific variation in the neurotoxic activity of B. neuwiedii venoms, whereas myotoxic activity is less variable.
- Published
- 2002
3. Histopathological changes in avian kidney caused by Bothrops insularis (jararaca ilhoa) venom and a phospholipase A2-containing fraction
- Author
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da Cruz Hoflingl, M.A., Paronetto, C.C.L., Cogo, J.C., Rodrigues-Simioni, L., and D'Abreu, A.C.F.
- Subjects
Avian ,6 - Ciencias aplicadas::61 - Medicina::615 - Farmacología. Terapéutica. Toxicología. Radiología [CDU] ,Kidney ,complex mixtures - Abstract
The histopathological changes induced in avian kidney by the intramuscular injection of Bothrops insularis (jararaca ilhda) venom and its phospholipase A2 (PLA2)-containing fraction were examined. Acute experiments (3 h and 24 h) with B. insularis crude venom (20 pg and 80 pg) or its PLA2-contaning fraction (10 pg and 40 pg) resulted in significant structural damage to the kidneys of 5-12-day-old chicks. Histopathological analysis indicated that the venom and its fraction acted on the renal tubules and glomeruli. The morphological changes, although widespread, varied in intensity from cell to cell, and from tubule to tubule in venom-injected chicks. The tubular and glomerular changes produced by the venom and its PLA2-containing fraction may be the result of a direct cytotoxic effect potentiated by ischemia-related disturbances in the regional hemodynamics. The venom and its fraction affected more segments along reptilian-type nephrons than along mammalian ones. This divergent sensitivity to the venom and its fraction may reflect the speciesspecific characteristics of B. insularis snake, an example of geographical isolation influencing its diet which is almost exclusively avian.
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- 2001
4. The pharmacological effect of Bothrops neuwiedii pauloensis (jararaca-pintada) snake venom on avian neuromuscular transmission
- Author
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Borja-Oliveira, C.R., primary, Durigon, A.M., additional, Vallin, A.C.C., additional, Toyama, M.H., additional, Souccar, C., additional, Marangoni, S., additional, and Rodrigues-Simioni, L., additional
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- 2003
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5. Rabbit antivenom efficacy against myotoxic and neurotoxic activities of Bothrops jararacussu venom and bothropstoxin-I
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Oshima-Franco, Y., primary, Leite, G. B., additional, Valério, A. A., additional, Hyslop, S., additional, Andriao-Escarso, S., additional, Giglio, J. R., additional, Prado-Franceschi, J., additional, Cruz-Höfling, M. A., additional, and Rodrigues-Simioni, L., additional
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- 2002
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6. INTRASPECIFIC VARIATION IN NEUROTOXIC AND MYOTOXIC ACTIVITIES OF Bothrops neuwiedii VENOMS
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BORJA-OLIVEIRA, C. R., primary, SOARES, A. M., additional, ZAMUNÉR, S. R., additional, HYSLOP, S., additional, GIGLIO, J. R., additional, PRADO-FRANCESCHI, J., additional, and RODRIGUES-SIMIONI, L., additional
- Published
- 2002
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- View/download PDF
7. Biological characterization of Bothrops marajoensis snake venom
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Walter Cavalcante, Hernandez-Oliveira, S., Galbiatti, C., Randazzo-Moura, P., Rocha, T., Ponce-Soto, L., Marangoni, S., Pai-Silva, M. D., Gallacci, M., Da Cruz-Höfling, M. A., Rodrigues-Simioni, L., Universidade Estadual de Campinas (UNICAMP), and Universidade Estadual Paulista (Unesp)
- Subjects
Marajó lancehead ,Neurotoxicity ,Neuromuscular junction ,Myotoxicity ,Presynaptic effects - Abstract
Made available in DSpace on 2016-07-07T12:36:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2011. Added 1 bitstream(s) on 2016-07-07T12:45:29Z : No. of bitstreams: 1 ISSN2044-0324-2011-02-37-41.pdf: 943442 bytes, checksum: de5846986ac9d173f5d2dba31d6ce61c (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) This study describes the effects of Bothrops marajoensis venom (Marajó lancehead) on isolated neuromuscular preparations of chick biventer cervicis (CBC) and mouse phrenic nerve-diaphragm (PND). At low concentrations (1µg/ml for CBC and 5µg/ml for PND), the venom exhibited a neuromuscular blocking without any damaging effect on the muscle integrity. At higher concentration (20µg/ml for PND), together with the neuromuscular blockade, there was a moderate myonecrosis. The results show differences between mammalian and avian preparations in response to venom concentration; the avian preparation was more sensitive to venom neurotoxic effect than the mammalian preparation. The possible presynaptic mechanism underlying the neuromuscular blocking effect was reinforced by the observed increase in MEPPs at the same time (at 15min) when the facilitation of twitch tension occurred. These results indicate that the B. marajoensis venom produced neuromuscular blockade, which appeared to be presynaptic at low concentrations with a postsynaptic component at high concentrations, leading to muscle oedema. These observations demand the fractionation of the crude venom and characterization of its active components for a better understanding of its biological dynamics. Universidade Estadual de Campinas, Departamento de Farmacologia, Faculdade de Ciências Médicas Universidade Estadual de Campinas, Departamento de Histologia e Embriologia, Instituto de Biologia Universidade Estadual de Campinas, Departamento de Bioquímica, Instituto de Biologia Universidade Estadual Paulista, Departamento de Morfologia, Instituto de Biociências de Botucatu Universidade Estadual Paulista, Departamento de Farmacologia, Instituto de Biociências de Botucatu
8. Cordia salicifolia and Lafoensia pacari plant extracts against the local effects of Bothrops jararacussu and Philodryas olfersii snake venoms.
- Author
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Schezaro-Ramos R, Collaço RC, Cogo JC, Dal-Belo CA, Rodrigues-Simioni L, Rocha T, and Randazzo-Moura P
- Abstract
Philodryas olfersii produces similar local effects to Bothrops jararacussu snakebite, which can induce misidentification and bothropic antivenom administration. Antivenom therapy is effective, but has its limitations regarding local damage. Since plants are used in folk medicine to treat snakebite victims, we evaluated the protective properties of Cordia salicifolia and Lafoensia pacari extracts against Philodryas olfersii and Bothrops jararacussu venoms. Preparations pretreated with both extracts inhibited > 90% the B. jararacussu venom-induced neuromuscular blockade, and 52% to 81% the P. olfersii venom-induced blockade. C. salicifolia inhibited the myonecrosis promoted by both venoms; however, L. pacari prevented only the myofilaments hypercontraction. Regarding haemorrhagic activity, C. salicifolia was more effective against B. jararacussu venom, while L. pacari was more effective against P. olfersii venom. On the other hand, for oedema-forming activity the results were the opposite. Considering that both extracts prevented (to different levels) the main manifestations of both snakebites (local symptoms), we endorse further studies involving these plants as coadjuvant in snakebite therapeutics., (© Copyright The Author(s).)
- Published
- 2020
9. Inhibition of Kv2.1 Potassium Channels by MiDCA1, A Pre-Synaptically Active PLA 2 -Type Toxin from Micrurus dumerilii carinicauda Coral Snake Venom.
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Schütter N, Barreto YC, Vardanyan V, Hornig S, Hyslop S, Marangoni S, Rodrigues-Simioni L, Pongs O, and Dal Belo CA
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- Animals, Cells, Cultured, Ganglia, Spinal cytology, Ganglia, Spinal physiology, Kv1.2 Potassium Channel genetics, Kv1.2 Potassium Channel physiology, Male, Mice, Inbred C57BL, Neurons drug effects, Neurons physiology, Oocytes physiology, Phospholipases A2, Xenopus, Coral Snakes, Elapid Venoms toxicity, Kv1.2 Potassium Channel antagonists & inhibitors, Potassium Channel Blockers toxicity
- Abstract
MiDCA1, a phospholipase A
2 (PLA2 ) neurotoxin isolated from Micrurus dumerilii carinicauda coral snake venom, inhibited a major component of voltage-activated potassium (Kv) currents (41 ± 3% inhibition with 1 μM toxin) in mouse cultured dorsal root ganglion (DRG) neurons. In addition, the selective Kv2.1 channel blocker guangxitoxin (GxTx-1E) and MiDCA1 competitively inhibited the outward potassium current in DRG neurons. MiDCA1 (1 µM) reversibly inhibited the Kv2.1 current by 55 ± 8.9% in a Xenopus oocyte heterologous system. The toxin showed selectivity for Kv2.1 channels over all the other Kv channels tested in this study. We propose that Kv2.1 channel blockade by MiDCA1 underlies the toxin's action on acetylcholine release at mammalian neuromuscular junctions.- Published
- 2019
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10. Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison.
- Author
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Rostelato-Ferreira S, André Dal Belo C, Ismael da Silva Junior P, Hyslop S, Rodrigues-Simioni L, and Augusto Alves Rocha-E-Silva T
- Abstract
Purpose: Rhinella schneideri is a toad found in many regions of the South America. The poison of the glands has cardiotoxic effect in animals and neuromuscular effects in mice and avian preparation. The purpose of this work was to identify the toxin responsible for the neuromuscular effect in avian and mice neuromuscular preparation. Methods: The methanolic extract from R. schneideri poison was fractioned by reversed phase HPLC. The purity and molecular mass were determined by LC/MS mass spectrometry. Chick biventer cervicis and mouse phrenic-nerve diaphragm were used as neuromuscular preparations to identify the toxin. Results: The purification resulted in 32 fractions, which 4 of them were active in neuromuscular preparation. The toxin of fraction 20 were chosen for better reproducibility of the whole extract activity and its molecular mass was 730.6 Da. The toxin produced facilitation of the muscle contraction followed by a complete neuromuscular blockade in chick biventer cervicis preparation in 90 min without interfering with the exogenous response to ACh and KCl. The quantal content was increased from 128 ± 13 (control) to 216 ± 44 (after 5 min and sustained until 60 min) in the presence of the toxin. Conclusion: In conclusion, our results demonstrated that the neuromuscular action of the poison of Rhinella schneideri is a multitoxin effect. More, the present work first isolated a 730.6 Da toxin that better represent the whole poison neuromuscular effect, to which is attributed a presynaptic action in avian and mouse neuromuscular preparation.
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- 2018
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11. Presynaptic Proteins as Markers of the Neurotoxic Activity of BmjeTX-I and BmjeTX-II Toxins from Bothrops marajoensis (Marajó Lancehead) Snake Venom.
- Author
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Lisboa A, Melaré R, Franco JR, Bis CV, Gracia M, Ponce-Soto LA, Marangoni S, Rodrigues-Simioni L, da Cruz-Höfling MA, and Rocha T
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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.
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- 2016
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12. Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom.
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Sucasaca-Monzón G, Randazzo-Moura P, Rocha T, Torres-Huaco FD, Vilca-Quispe A, Ponce-Soto LA, Marangoni S, da Cruz-Höfling MA, and Rodrigues-Simioni L
- Abstract
A new PLA2 (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on μ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homology with basic Asp49 PLA2 myotoxins from other Bothrops venoms. Bp-13 showed allosteric enzymatic behavior and maximal activity at pH 8.1, 36°-45°C. Full Bp-13 PLA2 activity required Ca(2+); its PLA2 activity was inhibited by Mg(2+), Mn(2+), Sr(2+), and Cd(2+) in the presence and absence of 1 mM Ca(2+). In the mouse phrenic nerve-diaphragm (PND) preparation, the time for 50% paralysis was concentration-dependent (P < 0.05). Both the replacement of Ca(2+) by Sr(2+) and temperature lowering (24°C) inhibited the Bp-13 PLA2-induced twitch-tension blockade. Bp-13 PLA2 inhibited the contractile response to direct electrical stimulation in curarized mouse PND preparation corroborating its contracture effect. In biventer cervicis preparations, Bp-13 induced irreversible twitch-tension blockade and the KCl evoked contracture was partially, but significantly, inhibited (P > 0.05). The main effect of this new Asp49 PLA2 of Bothrops pauloensis venom is on muscle fiber sarcolemma, with avian preparation being less responsive than rodent preparation. The study enhances biochemical and pharmacological characterization of B. pauloensis venom.
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- 2015
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13. Presynaptic neuromuscular action of a methanolic extract from the venom of Rhinella schneideri toad.
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Rostelato-Ferreira S, Dal Belo CA, Leite GB, Hyslop S, and Rodrigues-Simioni L
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Background: Rhinella schneideri, previously known as Bufo paracnemis, is a common toad in many regions of Brazil. Its venom exerts important cardiovascular effects on humans and other animals. Although this toad venom has been the subject of intense investigations, little is known about its neuromuscular activity., Methods: The neurotoxicity of a methanolic extract of R. schneideri venom was tested on mouse phrenic nerve-diaphragm (PND) preparations mounted for conventional twitch tension recording - in response to indirect stimulation - and for electrophysiological measurements., Results: Venom extract (50 μg/mL) increased the muscle twitch tension in PND preparations but did not significantly alter the resting membrane potential values. Electrophysiological evaluations showed that the extract (50 μg/mL) significantly augmented the frequency of miniature end-plate potential (from 38 ± 3.5 to 88 ± 15 after 60 minutes; n = 5; p < 0.05) and quantal content (from 128 ± 13 to 272 ± 34 after five minutes; n = 5; p < 0.05). Pretreatment with ouabain (1 μg/mL) for five minutes prevented the increase in quantal content (117 ± 18 and 154 ± 33 after five and 60 minutes, respectively)., Conclusion: These results indicate that the methanolic extract of R. schneideri venom acts primarily presynaptically to enhance neurotransmitter release in mouse phrenic-diaphragm preparations.
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- 2014
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14. Neuromuscular activity of Bothrops fonsecai snake venom in vertebrate preparations.
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Fernandes CT, Giaretta VM, Prudêncio LS, Toledo EO, da Silva IR, Collaço RC, Barbosa AM, Hyslop S, Rodrigues-Simioni L, and Cogo JC
- Abstract
The neuromuscular activity of venom from Bothrops fonsecai, a lancehead endemic to southeastern Brazil, was investigated. Chick biventer cervicis (CBC) and mouse phrenic nerve-diaphragm (PND) preparations were used for myographic recordings and mouse diaphragm muscle was used for membrane resting potential (RP) and miniature end-plate potential (MEPP) recordings. Creatine kinase release and muscle damage were also assessed. In CBC, venom (40, 80 and 160μg/ml) produced concentration- and time-dependent neuromuscular blockade (50% blockade in 85±9 min and 73±8 min with 80 and 160μg/ml, respectively) and attenuated the contractures to 110μM ACh (78-100% inhibition) and 40mM KCl (45-90% inhibition). The venom-induced decrease in twitch-tension in curarized, directly-stimulated preparations was similar to that in indirectly stimulated preparations. Venom (100 and 200μg/ml) also caused blockade in PND preparations (50% blockade in 94±13 min and 49±8 min with 100 and 200μg/ml, respectively) but did not alter the RP or MEPP amplitude. In CBC, venom caused creatine kinase release and myonecrosis. The venom-induced decrease in twitch-tension and in the contractures to ACh and K(+) were abolished by preincubating venom with commercial antivenom. These findings indicate that Bothrops fonsecai venom interferes with neuromuscular transmission essentially through postsynaptic muscle damage that affects responses to ACh and KCl. These actions are effectively prevented by commercial antivenom.
- Published
- 2014
15. Neuromuscular activity of Micrurus laticollaris (Squamata: Elapidae) venom in vitro.
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Carbajal-Saucedo A, Floriano RS, Dal Belo CA, Olvera-Rodríguez A, Alagón A, and Rodrigues-Simioni L
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- Animals, Chickens, Elapidae, In Vitro Techniques, Male, Membrane Potentials drug effects, Mice, Muscle, Skeletal drug effects, Neuromuscular Junction metabolism, Phrenic Nerve drug effects, Receptors, Nicotinic drug effects, Receptors, Nicotinic metabolism, Synaptic Transmission drug effects, Elapid Venoms pharmacology, Neuromuscular Junction drug effects
- Abstract
In this work, we have examined the neuromuscular activity of Micrurus laticollaris (Mexican coral snake) venom (MLV) in vertebrate isolated nerve-muscle preparations. In chick biventer cervicis preparations, the MLV induced an irreversible concentration- and time-dependent (1-30 µg/mL) neuromuscular blockade, with 50% blockade occurring between 8 and 30 min. Muscle contractures evoked by exogenous acetylcholine were completely abolished by MLV, whereas those of KCl were also significantly altered (86% ± 11%, 53% ± 11%, 89% ± 5% and 89% ± 7% for one, three, 10 and 30 µg of venom/mL, respectively; n = 4; p < 0.05). In mouse phrenic nerve-diaphragm preparations, MLV (1-10 µg/mL) promoted a slight increase in the amplitude of twitch-tension (3 µg/mL), followed by neuromuscular blockade (n = 4); the highest concentration caused complete inhibition of the twitches (time for 50% blockade = 26 ± 3 min), without exhibiting a previous neuromuscular facilitation. The venom (3 µg/mL) induced a biphasic modulation in the frequency of miniature end-plate potentials (MEPPs)/min, causing a significant increase after 15 min, followed by a decrease after 60 min (from 17 ± 1.4 (basal) to 28 ± 2.5 (t15) and 12 ± 2 (t60)). The membrane resting potential of mouse diaphragm preparations pre-exposed or not to d-tubocurarine (5 µg/mL) was also significantly less negative with MLV (10 µg/mL). Together, these results indicate that M. laticollaris venom induces neuromuscular blockade by a combination of pre- and post-synaptic activities.
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- 2014
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16. In vitro antiophidian mechanisms of Hypericum brasiliense choisy standardized extract: quercetin-dependent neuroprotection.
- Author
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Dal Belo CA, Lucho AP, Vinadé L, Rocha L, Seibert França H, Marangoni S, and Rodrigues-Simioni L
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- Animals, Brain cytology, Brain drug effects, Cell Survival drug effects, Crotalid Venoms toxicity, Crotoxin toxicity, Diaphragm drug effects, Humans, Hypericum chemistry, Mice, Phrenic Nerve physiopathology, Plant Extracts chemistry, Quercetin administration & dosage, Neuromuscular Blockade, Neuroprotective Agents administration & dosage, Phrenic Nerve drug effects, Plant Extracts administration & dosage
- Abstract
The neuroprotection induced by Hypericum brasiliense Choisy extract (HBE) and its main active polyphenol compound quercetin, against Crotalus durissus terrificus (Cdt) venom and crotoxin and crotamine, was enquired at both central and peripheral mammal nervous system. Cdt venom (10 μg/mL) or crotoxin (1 μg/mL) incubated at mouse phrenic nerve-diaphragm preparation (PND) induced an irreversible and complete neuromuscular blockade, respectively. Crotamine (1 μg/mL) only induced an increase of muscle strength at PND preparations. At mouse brain slices, Cdt venom (1, 5, and 10 μg/mL) decreased cell viability. HBE (100 μg/mL) inhibited significantly the facilitatory action of crotamine (1 μg/mL) and was partially active against the neuromuscular blockade of crotoxin (1 μg/mL) (data not shown). Quercetin (10 μg/mL) mimicked the neuromuscular protection of HBE (100 μg/mL), by inhibiting almost completely the neurotoxic effect induced by crotoxin (1 μg/mL) and crotamine (1 μg/mL). HBE (100 μg/mL) and quercetin (10 μg/mL) also increased cell viability in mice brain slices. Quercetin (10 μg/mL) was more effective than HBE (100 μg/mL) in counteracting the cell lysis induced by Cdt venom (1 and 10 μg/mL, resp.). These results and a further phytochemical and toxicological investigations could open new perspectives towards therapeutic use of Hypericum brasiliense standardized extract and quercetin, especially to counteract the neurotoxic effect induced by snake neurotoxic venoms.
- Published
- 2013
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17. Biochemical, pharmacological, and structural characterization of new basic PLA2 Bbil-TX from Bothriopsis bilineata snake venom.
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Corasolla Carregari V, Stuani Floriano R, Rodrigues-Simioni L, Winck FV, Baldasso PA, Ponce-Soto LA, and Marangoni S
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- Amino Acid Sequence, Animals, Calcium metabolism, Cell Line, Edema pathology, Hydrogen-Ion Concentration, Hydrolysis, Inflammation, Interleukin-1 metabolism, Interleukin-6 metabolism, Mass Spectrometry, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Phospholipases A2 pharmacology, Phospholipases A2, Secretory pharmacology, Reptilian Proteins pharmacology, Spectrometry, Mass, Electrospray Ionization, Tumor Necrosis Factor-alpha metabolism, Bothrops, Phospholipases A2 chemistry, Phospholipases A2, Secretory chemistry, Reptilian Proteins chemistry, Snake Venoms enzymology
- Abstract
Bbil-TX, a PLA2, was purified from Bothriopsis bilineata snake venom after only one chromatographic step using RP-HPLC on μ-Bondapak C-18 column. A molecular mass of 14243.8 Da was confirmed by Q-Tof Ultima API ESI/MS (TOF MS mode) mass spectrometry. The partial protein sequence obtained was then submitted to BLASTp, with the search restricted to PLA2 from snakes and shows high identity values when compared to other PLA2s. PLA2 activity was presented in the presence of a synthetic substrate and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0 and 25-37°C. Maximum PLA2 activity required Ca(2+) and in the presence of Cd(2+), Zn(2+), Mn(2+), and Mg(2+) it was reduced in the presence or absence of Ca(2+). Crotapotin from Crotalus durissus cascavella rattlesnake venom and antihemorrhagic factor DA2-II from Didelphis albiventris opossum sera under optimal conditions significantly inhibit the enzymatic activity. Bbil-TX induces myonecrosis in mice. The fraction does not show a significant cytotoxic activity in myotubes and myoblasts (C2C12). The inflammatory events induced in the serum of mice by Bbil-TX isolated from Bothriopsis bilineata snake venom were investigated. An increase in vascular permeability and in the levels of TNF-a, IL-6, and IL-1 was was induced. Since Bbil-TX exerts a stronger proinflammatory effect, the phospholipid hydrolysis may be relevant for these phenomena.
- Published
- 2013
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18. Guanidine affects differentially the twitch response of diaphragm, extensor digitorum longus and soleus nerve-muscle preparations of mice.
- Author
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Ferrari R, Rodrigues-Simioni L, and da Cruz Höfling MA
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- Animals, Calcium metabolism, Diaphragm cytology, Diaphragm metabolism, Male, Mice, Muscle, Skeletal cytology, Muscle, Skeletal metabolism, Myography, Synaptic Transmission drug effects, Diaphragm drug effects, Guanidine pharmacology, Muscle Contraction drug effects, Muscle, Skeletal drug effects, Neuromuscular Junction drug effects
- Abstract
Guanidine has been used with some success to treat myasthenia gravis and myasthenic syndrome because it increases acetylcholine release at nerve terminals through K⁺, Na⁺ and Ca²⁺ channels-involving mechanisms. Currently, guanidine derivatives have been proposed for treatment of several diseases. Studies aimed at providing new insights to the drug are relevant. Experimentally, guanidine (10 mM) induces on mouse phrenic nerve-diaphragm (PND) preparations neurotransmission facilitation followed by blockade and a greatest secondary facilitation after its removal from bath. Herein, we hypothesized that this peculiar triphasic response may differ in muscles with distinct twitch/metabolic characteristics. Morphological alterations and contractile response of PND, extensor digitorum longus (EDL) and soleus (SOL) preparations incubated with guanidine (10 mM) for 15, 30, 60 min were analyzed. Guanidine concentrations of 5 mM (for PND and EDL) and 1 mM (for EDL) were also tested. Guanidine triphasic effect was only observed on PND regardless the concentration. The morphological alterations in muscle tissue varied along time but did not impede the PND post-wash facilitation. Higher doses (20-25 mM) did not increase EDL or SOL neurotransmission. The data suggest a complex mechanism likely dependent on the metabolic/contractile muscle phenotype; muscle fiber types and density/type of ion channels, sarcoplasmic reticulum and mitochondria organization may have profound impact on the levels and isoform expression pattern of Ca²⁺ regulatory membrane proteins so reflecting regulation of calcium handling and contractile response in different types of muscle.
- Published
- 2012
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19. Presynaptic effect of a methanolic extract of toad (Rhinella schneideri) poison in avian neuromuscular preparation.
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Rostelato-Ferreira S, Dal Belo CA, da Cruz-Höfling MA, Hyslop S, and Rodrigues-Simioni L
- Abstract
The neurotoxicity of a methanolic extract of toad (Rhinella schneideri) poison was examined in chick biventer cervicis preparations. The methanolic extract (1, 3, 10 and 30µg/ml) caused concentration-dependent blockade at the three highest concentrations (time for 50% blockade, mean±SEM: 84±10, 51±3 and 12±0.8min for 3, 10 and 30µg/ml, respectively; n=6-8 each) that was preceded by significant, transient facilitation at 10μg/ml. Contractures to exogenous ACh (110μM) or KCl (20mM) were unaffected by the blockade. In curarized (d-Tc, 1μg/ml) preparations, the extract (10µg/ml) caused complete, irreversible blockade that persisted after extensive washing. The extract did not significantly alter the creatine kinase release or morphology of biventer cervicis muscle. These results indicate that the methanolic extract of R. schneideri poison acts primarily presynaptically to enhance neurotransmitter release in this avian preparation.
- Published
- 2011
20. Biological characterization of Bothrops marajoensis snake venom.
- Author
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Cavalcante WL, Hernandez-Oliveira S, Galbiatti C, Randazzo-Moura P, Rocha T, Ponce-Soto L, Marangoni S, Pai-Silva MD, Gallacci M, da Cruz-Höfling MA, and Rodrigues-Simioni L
- Abstract
This study describes the effects of Bothrops marajoensis venom (Marajó lancehead) on isolated neuromuscular preparations of chick biventer cervicis (CBC) and mouse phrenic nerve-diaphragm (PND). At low concentrations (1µg/ml for CBC and 5µg/ml for PND), the venom exhibited a neuromuscular blocking without any damaging effect on the muscle integrity. At higher concentration (20μg/ml for PND), together with the neuromuscular blockade, there was a moderate myonecrosis. The results show differences between mammalian and avian preparations in response to venom concentration; the avian preparation was more sensitive to venom neurotoxic effect than the mammalian preparation. The possible presynaptic mechanism underlying the neuromuscular blocking effect was reinforced by the observed increase in MEPPs at the same time (at 15min) when the facilitation of twitch tension occurred. These results indicate that the B. marajoensis venom produced neuromuscular blockade, which appeared to be presynaptic at low concentrations with a postsynaptic component at high concentrations, leading to muscle oedema. These observations demand the fractionation of the crude venom and characterization of its active components for a better understanding of its biological dynamics.
- Published
- 2011
21. The neuromuscular activity of Micrurus pyrrhocryptus venom and its neutralization by commercial and specific coral snake antivenoms.
- Author
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Camargo TM, de Roodt AR, da Cruz-Höfling MA, and Rodrigues-Simioni L
- Abstract
The neuromuscular activity ofMicrurus pyrrochryptus venom was studied in chick biventer cervicis (BC) and mouse phrenic nerve-diaphragm (PND) preparations. The venom (0.5-50μg/ml) caused irreversible, time- and concentration-dependent blockade, with BC being more sensitive than PND (50% blockade with 10μg/ml in 22±;3min and 62±4min, respectively; mean±SEM, n=6; p<0.05). In BC preparations, venom (0.5μg/ml) progressively abolished ACh-induced contractures, whereas contractures to exogenous KCl and muscle twitches in curarized preparations were unaffected. The venom neither altered creatine kinase release (venom: 25.8±1.75IU/l vs control: 24.3±2.2IU/l, n=6, after 120min), nor it caused significant muscle damage (50μg of venom/ml vs control: 3.5±0.8% vs 1.1±0.7% for PND; 4.3±1.5% vs 1.2±0.5% for BC, n=5). The venom had low PLA(2) activity. Neurotoxicity was effectively neutralized by commercial Micrurus antivenom and specific antivenom. These findings indicate that M. pyrrhocryptus venom acts postsynaptically on nicotinic receptors, with no significant myotoxicity.
- Published
- 2011
22. Heparin at low concentration acts as antivenom against Bothrops jararacussu venom and bothropstoxin-I neurotoxic and myotoxic actions.
- Author
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Rostelato-Ferreira S, Leite GB, Cintra AC, da Cruz-Höfling MA, Rodrigues-Simioni L, and Oshima-Franco Y
- Abstract
Heparin has been shown to antagonize myotoxic effects of crotaline venoms. Here a very low heparin concentration (LHC) was examined in its ability to antagonize the neurotoxic/myotoxic effects of Bothrops jararacussu venom and its phospholipase A(2) myotoxin, bothropstoxin-I (BthTX-I), in an in vitroz nerve-muscle preparation and in mice gastrocnemius. Normalization of results was done by assays with commercial antibothropic antivenom (CBA). LHC (1IU/ml) added to the incubation bath reduced by 4- and 4.5-fold (vs 2.8- and 2.5-fold by CBA) the neuromuscular paralysis, by 5.4 and 4.4-fold (vs 2.5- and 13.3-fold by CBA) the percentage of fibers damaged and by 6- and 1.7-fold (vs 30- and 1.6-fold by CBA) the CK activity induced by B. jararacussu and BthTX-I, respectively. Protamine sulphate added 15min after the incubation of the preparation with LHC+venom, avoided the LHC neutralizing effect against venom neurotoxicity. This strongly attests that given the polycationic nature of protamine, it probably complexed with the polyanionic heparin making it unattainable for binding to basic components of venom, reducing toxicity. Since heparin antagonism is generally stronger against venom effects than is myotoxin we discuss that other venom components than the BthTX-I are likely target for the antagonism promoted by the polyanionic heparin.
- Published
- 2010
23. Influence of local anesthetics on the neuromuscular blockade produced by rocuronium: effects of lidocaine and 50% enantiomeric excess bupivacaine on the neuromuscular junction.
- Author
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Braga Ade F, Carvalho VH, Braga FS, Rodrigues-Simioni L, Loyola YC, and Potério GB
- Subjects
- Animals, Bupivacaine pharmacology, In Vitro Techniques, Lidocaine pharmacology, Male, Neuromuscular Nondepolarizing Agents administration & dosage, Rats, Rats, Wistar, Rocuronium, Androstanols administration & dosage, Anesthetics, Local administration & dosage, Anesthetics, Local pharmacology, Bupivacaine administration & dosage, Lidocaine administration & dosage, Neuromuscular Blockade, Neuromuscular Junction drug effects
- Abstract
Background and Objectives: The effects of local anesthetics (LA) on neuromuscular transmission and their influence on the neuromuscular blockade produced by competitive neuromuscular blockers have not been fully investigated. The objective of this study was to evaluate, in vitro, the effects of lidocaine and 50% enantiomeric excess bupivacaine (S75-R25) on the neuromuscular blockade produced by rocuronium., Methods: The rats were divided in five groups (n = 5) according to the drug used: isolated lidocaine, bupivacaine (S75-R25), or rocuronium (groups I, II, and II); and rocuronium in preparations previously exposed to LAs (groups IV and V). The concentrations used were as follows: 20 microg x mL(-1), 5 microg x mL(-1), and 4 microg x mL(-1) of lidocaine, bupivacaine (S75-R25), and rocuronium, respectively. The following parameters were evaluated: 1) the strength of muscular contraction of the diaphragm to indirect electrical stimulations, before and 60 minutes after the isolated addition of the LAs and rocuronium, and the association AL-rocuronium; and 2) the effects of LAs on membrane potential (MP) and miniature end-plate potentials (MEPP). The effect of LAs on muscle contraction in response to acetylcholine was evaluated in chick biventer cervicis preparations., Results: Isolated lidocaine and bupivacaine (S75-R25) did not change the muscular response and the levels of MPs. In preparations exposed to LAs, rocuroniuminduced blockade was significantly greater than that produced by rocuronium alone. In chick biventer cervicis preparations, lidocaine and bupivacaine (S75R25) decreased contraction in response to acetylcholine. Lidocaine increased the frequency of MEPPs, which was followed by the blockade; bupivacaine (S75R25) caused a reduction in MEPPs followed by blockade., Conclusions: Local anesthetics caused a potentiation of the neuromuscular blockade produced by rocuronium. The results showed pre- and post-synaptic effects.
- Published
- 2009
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