Characterization Of The Crotalus Durissus Terrificus Venom By Atomic Force Microscopy

A preliminary analysis of the morphology of crude CDT venom was performed using atomic force microscopy (AFM). As a result, a direct measurement of the forces between the surface of the venom crystals and a sharp probe tip was achieved.20413171321Azevedo Marques, M.M., Cupo, P., Coimbra, T.M., Hering, S.E., Rossi, M.A., Laure, C.J., (1985) Toxicon, 23, p. 631Sanchez, E.E., Soliz, L.A., Susana Ramirez, M., Perez, J.C., (2001) Toxicon, 39, p. 523Soares, A.M., Mancin, A.C., Cecchini, A.L., Arantes, E.C., Franca, S.C., Gutierrez, J.M., Giglio, J.R., (2001) Int. J. Biochem. Cell Biol., 33, p. 877Choumet, V., Lafaye, P., Demangel, C., Bon, C., Mazie, J.C., (1999) Biol. Chem., 380, p. 561Sanchez, S.A., Chen, Y., Muller, J.D., Gratton, E., Hazlett, T.L., (2001) Biochemistry, 40, p. 6903Sharma, S., Jabeen, T., Singh, J.K., Bredhorst, R., Vogel, C.W., Betzel, C., Singh, T.P., (2001) Acta Crystallogr., Sect. D: Biol. Crystallogr., 57, p. 596Nonato, M.C., Garrat, R.C., Mascarenhas, Y.P., Jesus, W.D.P., Assakura, M.T., Serrano, S.M.T., Oliva, G., (2001) Biol. Crystallogr., D, 57, p. 599Arni, R.K., Ward, R.J., (1996) Toxicon, 34, p. 827Souza, D.H.F., Selistre-deAraujo, H.S., Garratt, R.C., (2000) Toxicon, 38, p. 1307Binning, G., Coate, C.F., Gerber, G., (1986) Phys. Rev. Lett., 56, p. 930Baranauskas, V., Vidal, B.C., Parizotto, N.A., (1998) Appl. Biochem. Biotechnol., 69, p. 91Baranauskas, V., Garavello-Freitas, I., Jingguo, Z., Cruz-Hofling, M.A., (2001) J. Vac. Sci. Technol. A, 19, p. 104

Expression Of Vegf And Flk-1 And Flt-1 Receptors During Blood-brain Barrier (bbb) Impairment Following Phoneutria Nigriventer Spider Venom Exposure

Apart from its angiogenic and vascular permeation activity, the vascular endothelial growth factor (VEGF) has been also reported as a potent neuronal protector. Newborn rats with low VEGF levels develop neuron degeneration, while high levels induce protective mechanisms in several neuropathological conditions. Phoneutria nigriventer spider venom (PNV) disrupts the blood-brain barrier (BBB) and causes neuroinflammation in central neurons along with excitotoxic signals in rats and humans. All these changes are transient. Herein, we examined the expression of VEGF and its receptors, Flt-1 and Flk-1 in the hippocampal neurons following envenomation by PNV. Adult and neonatal rats were evaluated at time limits of 2, 5 and 24 h. Additionally, BBB integrity was assessed by measuring the expression of occludin, β-catenin and laminin and neuron viability was evaluated by NeuN expression. VEGF, Flt-1 and Flk-1 levels increased in PNV-administered rats, concurrently with respective mRNAs. Flt-1 and Flk-1 immunolabeling was nuclear in neurons of hippocampal regions, instead of the VEGF membrane-bound typical location. These changes occurred simultaneously with the transient decreases in BBB-associated proteins and NeuN positivity. Adult rats showed more prominent expressional increases of the VEGF/Flt-1/Flk-1 system and earlier recovery of BBB-related proteins than neonates. We conclude that the reactive expressional changes seen here suggest that VEGF and receptors could have a role in the excitotoxic mechanism of PNV and that such role would be less efficient in neonate rats. © 2013 by the authors; licensee MDPI, Basel, Switzerland.51225722588Vassilevsky, A.A., Koslov, S.A., Egorov, T.A., Grishin, E.V., Purification and characterization of biologically active peptides from spider venoms (2010) Methods Mol. 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Bull., 73, pp. 114-126Cruz-Höfling, M.A., Rapôso, C., Verinaud, L., Zago, G.M., Neuroinflammationand astrocytic reaction in the course of Phoneutria nigriventer (armed-spider) blood-brain barrier (BBB) opening (2009) Neurotoxicology, 30, pp. 636-646Jin, K., Zhu, Y., Sun, Y., Mao, X.O., Xie, L., Greenberg, D.A., Vascular endothelial growth factor (VEGF) stimulates neurogenesis in vitro and in vivo (2002) Proc. Natl. Acad. Sci. USA, 99, pp. 11946-11950Ferrara, N., Gerber, H.P., le Couter, J., The biology of VEGF and its receptors (2003) Nat. Med., 9, pp. 669-676Semenza, G., Signal transductions to hypoxia-inducible factor 1 (2002) Biochem. 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Laser Effects On Osteogenesis

The traumatic or surgical cutting of a long bone is immediately followed by a sequence of repair processes in which the osteogenic cells of the periosteum start to proliferate and differentiate in osteoblast cells. In this work, we explored the influence of a He-Ne laser on osteogenesis after a controlled surgical fracture. We used young male adult Wistar rats (of mass between 250 and 300 g). The fracture was provoked by piercing a 2-mm-diameter hole in just one cortical tibia surface. Laser treatment was started 24 h after the surgery. The animals were separated into three groups, for different radiation doses, and after daily applications, they were sacrificed at 8 or 15 days. Light and electron microscopies revealed that the laser treatment of the lesion with doses of 31.5 and 94.7 J cm -2 resulted in the formation of thicker bony trabeculae, which indicates a greater synthesis of collagen fibers and therefore that the osteoblastic activity was increased by the low-energy laser radiation.154548554Lai, J.sm., Tham, C.C.Y., Lam, D.S.C., (1999) Eye, 13, p. 26Bauerberg, J.M., (1999) J. Refract. Surg., 15, p. 66McCarthy, D.C., (1998) Photonics Spectra, 32, p. 90Kitchen, S.S., Partridge, C.J., (1987) Physiotherapy, 77, p. 160Trelles, M.A., Mayayo, E., (1987) Laser Surg. Med., 7, p. 36Yaakobi, T., Maltz, L., Oron, U., (1996) Calcif. Tissue, 59, p. 207Luger, E.J., Rochkind, S., Wollman, Y., Kogan, G., Dekel, S., (1998) Laser Surg. Med., 22, p. 97Schmorl, G., (1922) Die Pathologisch-Histologische Untersuchungsmethoden, p. 243. , Leipzig, Germany: F.G.W. VogelDavid, R., Nissan, M., Cohen, I., Soudry, M., (1996) Laser Surg. Med., 19, p. 19Zaturunsky, B.M., (1988) Lasers Light Opthalmol., 2, p. 63Karu, T., (1989) Health Phys., 56, p. 691Friedmann, H., Lubart, R., Laulicht, I., Rochkind, S., (1991) J. Photochem. Photobiol., B, 11, p. 87Rochkind, S., Ouakine, G.E., (1992) Neurol. Res., 14, p.

Functional And Dissociation Properties And Structural Organization Of The Hemocyanin Of Ampullaria Canaliculata (gastropoda, Mollusca)

1. 1. The molecular weight of the native hemocyanin from Ampullaria canaliculata determined by gel filtration has a molecular mass of 7.5 × 106 Da. Dialysis against buffer free of Ca2+ and Mg2+ ions dissociated the hemocyanin into molecular species of 1.0 × 106 and 3.2 × 105 Da. 2. 2. The oxygen-binding curves of hemocyanin were sigmoidal with Hill coefficient of about 2. A positive Bohr effect was demonstrable at pH values ranging from 6.5 to 9.0. 3. 3. Differences between associated and dissociated hemocyanin as well as the effect of the divalent cations on the aggregation of hemocyanin molecule were illustrated by immunoelectrophoresis analysis. 4. 4. Negatively stained preparations of hemocyanin observed by TEM showed the cylindrical configuration of molluscan hemocyanins molecules. Its inner structure, enhanced by the negative contrasting, reveals a regular periodicity as a result of the alignment of serial subunits. © 1993.10503/04/15725730Barros, Propriedades fisico-quimicas e funcionais da hemocianina de Molusco Gastropoda Ampullaria canaliculata (1992) Dissertation Thesis, p. 58Burky, Burky, Buoyancy changes as related to respiratory behavior in an amphibious snail, Pomacea urcelus (Muller), from Venezuela (1977) The Nautilus, 91, pp. 97-104van Bruggen, Schutter, van Breemen, Bijlholt, Wichertjes, Arthropodan and Molluscan hemocyanins (1981) Electron Microscopy of Proteins, 1, p. 1938. , J.R. Harris, Academic Press, Londonvan Bruggen, An electron microscopists view of the quaternary structure of arthropodan and molluscan hemocyanins (1983) Life Chem. (Suppl.), 1, pp. 1-14Cruz-Hofling, Barròs, Matsuura, Focesi, Jr, Preliminary electron microscopic studies of hemocyanin from the Gastropod Ampullaria canaliculata (1992) Proceedings of the 1st Atlantic Congress of Electron Microscopy, , Merida, VenezuelaD'Enghien, Elliot, Bartels, van Bruggen, The haemocyanin of Pila leopoldvillensis—II. Quaternary structure and oxygenation (1971) Comp. Biochem. Physiol., 40 B, pp. 1045-4055Ellerton, Ellerton, Robinson, Hemocyanin—A current perspective (1983) Prog. biophys. molec. Biol., 41, pp. 143-248Fernández-Morán, van Bruggen, Ohtsuki, Macromolecular organization of hemocyanins and apohemocyanins as revealed by electron microscopy (1966) Journal of Molecular Biology, 16, pp. 191-207Hamilton, Herskovits, Furcinitti, Wall, Scanning transmission electron microscopic study of Molluscan hemocyanins in various aggregation states: comparison with light scattering molecular weights (1989) J. ultrastruct. molec. struct. Res., 102, pp. 221-228Heirwegh, Borgion, Lontie, Separation and absorption spectra of alpha and beta-hemocyanin of Helix pomatia (1961) Biochim. biophys. Acta, 48, pp. 517-526Herskovits, Recent aspects of the subunit organization and dissociation of hemocyanins (1988) Comp. Biochem. Physiol., 91 B, pp. 579-611Herskovits, Russell, Carberry, Light-scattering investigations of the subunit structure and sequencial dissociation of Homarus americanus hemocyanin (1984) Biochemistry, 23, pp. 1875-1881Herskovits, Blake, Gonzalez, Hamilton, Wall, Subunit structure and higher order assembly of the hemocyanins of the Melogenidae family: Melongena corona (Gmelin), Bysicon canaliculatum (Linne), B. carica (Gmelin), B. contrarium (Conrad), and B. spiratum (Lamarck) (1989) Comp. Biochem. Physiol., 94 B, pp. 415-421Herskovits, Otero, Hamilton, The hemocyanin of the Ramshorn snail, Marisa cornuarietis (Linne) (1990) Comp. Biochem. Physiol., 97 B, pp. 623-629Lamy, Immunological approach to the structure and evolution of hemocyanins (1982) EMBO Workshop, pp. 15-25. , Life ChemLarimer, Riggs, Properties of hemocyanins—I. The effect of calcium ions on the oxygen equilibrium of crayfish hemocyanin (1964) Comp. Biochem. Physiol., 13, pp. 35-46Laurent, Killander, A theory of gel filtration and its experimental verification (1964) Journal of Chromatography A, 14, pp. 317-330Mangun, Respiratory function of the hemocyanins (1980) Am. Zool., 20, pp. 19-38Mangun, Salt sensitivity of the hemocyanin of euri- and stenohaline squids (1991) Comparative Biochemistry and Physiology Part A: Physiology, 99 A, pp. 159-161Masseyeff, Gombert, Tanguy, Neuzil, Contribution a l'etude biochimique d'hémocyanine de Cymbium neptuni. II. Etude électrophoretique et immunoelectrophoretique (1963) Bull. Soc. chim. 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Experimental Studies Of Rat Injured Tibiae After Hene Radiation

Atomic force microscopy (AFM) and scanning electron microscopy (SEM) have been used to quantify bone morphology during post injury ossification in rat tibiae and characterize the differences induced by laser photostimulation compared with the naturally-occurring process. SEM and AFM images of the cortical bone at the recovery stages of 8 and 15 days after the injury are discussed.5287102107Lai, J.S.M., Tham, C.C.Y., Lam, D.S.C., (1999) Eye, 13, p. 26Bauerberg, J.M., (1999) J. Refract. Surg., 15, p. 66McCarthy, D.C., (1998) Photonics Spectra, 32, p. 90Baranauskas, V., Garavello-Freitas, I., Jingguo, Z., (2001) J. Vac. Sci. Technol. A, 19, p. 1042Garavello, I., Baranauskas, V., Joazeiro, P.P., Padovani, C.R., Dal Pai-Silva, M., Cruz-Höfling, M.A., J. Photochem. Photobiol. B., , (submitted)Garavello, I., Baranauskas, V., Cruz-Höfling, M.A., Histol. Histopathol., , (submitted)Barushka, O., Yaakobi, T., Oron, U., (1995) Bone, 16, p. 47Yaakobi, T., Maltz, L., Oron, U., (1996) Calcif. Tissue Int., 59, p. 29

Histoenzymological And Ultrastructural Changes In Lateral Muscle Fibers Of Oreochromis Niloticus (teleostei: Cichlidae) After Local Injection Of Veratrine

The effects of veratrine have been investigated in mammalian, amphibian, and crustacean muscle, but not in fish. In this work, the action of veratrine was studied in the lateral muscle of the freshwater teleost Oreochromis niloticus after intramuscular injection. Histoenzymological typing and electron microscopy of muscle fibers before and 15, 30, and 60 min after veratrine injection (10 ng/kg fish) were used to indirectly assess the morphological changes and the oxidative and m-ATPase activities. In some cases, muscles were pretreated with tetrodotoxin to determine whether the ultrastructural changes were the result of Na+ channel activation by veratrine. Veratrine altered the metabolism of fibers mainly after 30 min. Oxidative fibers showed decreased NADH-TR activity, whereas that of glycolytic and oxidative-glycolytic type fibers increased. There was no change in the m-ATPase activity of the three fiber types, except at 60 min postveratrine, when a novel fiber type, which showed no reversal after acidic and alkaline preincubations, appeared. Ultrastructural damage involved sarcomeres, myofibrils, and mitochondria, but the T-tubules remained intact. Pretreatment with tetrodotoxin (1 ng/ml) prevented the ultrastructural changes caused by veratrine. These results show that in fish skeletal muscle veratrine produces some effects that are not seen in mammalian muscle.1166525534Battersby, B.J., Moyes, C.D., Influence of acclimation temperature on mitochondrial DNA, RNA, and enzymes in skeletal muscle (1998) Am J Physiol, 275, pp. R905-R912Benforado, J.M., The veratrum alkaloids (1967) Physiological pharmacology, pp. 331-398. , Root WS, Hofmann, FG (eds). Academic Press, New YorkBrooke, M.H., Kaiser, K.K., Muscle fiber types: How many and what kind? (1970) Arch Neurol, 23, pp. 369-379Carpené, E., Veggetti, A., Increase in muscle fibers in the lateralis muscle (white portion) of Mugilidae (Pisces, Teleostei) (1981) Experientia, 37, pp. 191-193Celesia, G.G., Disorders of membrane channels or channelopathies (2001) Clin Neurophysiol, 112, pp. 2-18Cestèle, S., Catteral, W.A., Molecular mechanisms of neurotoxin action on voltage-gated sodium channels (2000) Biochimie, 82, pp. 883-892Cogswell, A.M., Stevens, R.J., Hood, D.A., Properties of skeletal muscle mitochondria isolated from subsarcolemmal and intermyofibrillar regions (1993) Am J Physiol, 264, pp. C383-C389Crockford, T., Johnston, I.A., Temperature acclimation and the expression of contractile protein isoforms in the skeletal muscles of the common carp (Cyprinus carpio L.) 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Mouse Extensor Digitorum Longus And Soleus Show Distinctive Ultrastructural Changes Induced By Veratrine

We investigated whether veratrine (5 μl, 10 ng/kg) injected into the mouse extensor digitorum longus (EDL) (fast-twitch) and soleus (SOL) (slow-twitch) muscles provokes distinctive ultrastructural disturbances 15, 30 and 60 min later. The mitochondria in SOL were affected earlier (within 15 min) than in EDL. Swelling of the sarcoplasmic reticulum terminal cisternae was more marked in EDL than in SOL and caused distortion of sarcomeres so that fragmentation of myofilaments was more pronounced in EDL. Hypercontracted sarcomeres were seen mainly in SOL and veratrine caused infoldings of the sarcolemma only in this muscle. In both muscles, the T-tubules remained unaffected and by 60 min after veratrine most of the above alterations had reverted to normal. Pretreatment with tetrodotoxin prevented the alterations induced by veratrine. This suggests that most of the alterations resulted from the enhanced influx of Na+ into muscle fibers. These results emphasize the importance of considering the type of muscle when studying the action of myotoxic agents.343305313Albuquerque, E.X., Warnick, J.E., Sansone, F.M., The pharmacology of batrachotoxin. II. Effect on electrical properties of the mammalian nerve and skeletal muscle membranes (1971) J. Pharmacol. Exp. Ther., 176, pp. 511-528Appell, H.J., Variability in microvascular pattern dependent upon muscle fiber composition (1984) Skeletal Muscle Microcirculation, p. 15. , Hammersen F. and Messmer K. eds., Karger, BaselBalnave, C.D., Allen, D.G., Evidence for Na+/Ca2- exchange in intact single skeletal muscle fibers from the mouse (1998) Am. J. Physiol., 274, pp. C940-C946Barcenas-Ruiz, L., Beuckelman, D.J., Wier, W.G., Sodium-calcium exchange in heart: Membrane currents and changes in [Ca2+] (1987) Science, 238, pp. 1720-1722Becker, S., Gordon, R.D., Sodium channel specific neurotoxins: Recent advances in the understanding of their molecular mechanisms (1992) Handbook of Experimental Pharmacology. Selective Neurotoxicity, pp. 719-737. , Herken H. and Hucho F. eds., Springer-Verlag, New York-BerlinBeeler, T., Osmotic changes of sarcoplasmic reticulum vesicles during Ca2+ uptake (1983) J. Memb. Biol., 76, pp. 165-171Benforado, J.M., The veratrum alkaloids (1967) Physiological Pharmacology, pp. 331-398. , Root W.S. and Hofmann F.G. eds., Academic Press, New YorkBlaustein, M.P., The interrelationship between sodium and calcium fluxes across cell membranes (1974) Rev. Physiol. Biochem. Pharmacol., 70, pp. 32-82Castillo, E., González-Serratos, H., Rasgado-Flores, H., Rozycka, M., Na+-Ca2+ exchange studies in frog phasic muscle cells (1991) Ann. N.Y. Acad. Sci., 639, pp. 554-557Catterall, W.A., Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes (1980) Ann. Rev. Pharmacol. Toxicol., 20, pp. 15-43Chang, C.C., Tseng, H.K., Effect of crotamine, a toxin of South American rattlesnake venom, on the sodium channel of murine skeletal muscle (1978) Br. J. Pharmacol., 63, pp. 551-559Chang, C.C., Hong, S.J., Su, M.J., A study on the membrane depolarization of skeletal muscles caused by a scorpion toxin, sea anemone toxin II and crotamine and the interaction between toxins (1983) Br. J. Pharmacol., 79, pp. 673-680Cruz-Höfling, M.A., Neder, F., Rodrigues-Simioni, L., Vital Brazil, O., Effects of veratrine on the ultrastructure of nerve terminals of the mouse diaphragm in vitro (1987) Proceedings of the XI Colloquium of the Brazilian Society for Electron Microscopy, p. 139. , Caxambu, MG, BrasilCruz-Höfling, M.A., Vital Brazil, O., Alterações ultraestruturais causadas pela veratrina no diafragma isolado de camundongo (1990) Proceedings of the V Annual Meeting of FESBE, p. 324. , Caxambu, MG, BrasilDauber, W., Zur Fasertypischen Morphologie und Funktion der Triaden im Skeletmuskel des Frosches (Rana sculenta) (1979) Z. Microsc. Anat. Forsch., 93, pp. 512-536Donoso, P., Hidalgo, C., Sodium-calaum exchange in transverse tubules isolated from frog skeletal muscle (1989) Biochim. Biophys. Acta, 978, pp. 8-16Eastwood, A.B., Franzini-Armstrong, C., Peracchia, C., Structure of membranes in crayfish muscle: Comparison of phasic and tonic fibers (1982) J. Muscle Res. Cell Motil., 3, pp. 273-294Endo, M., Calcium release from the sarcoplasmic reticulum (1977) Physiol. Rev., 57, pp. 71-108Fletcher, J.E., Hubert, M., Wieland, S.J., Gong, Q.-H., Jiang, M.-S., Similarities and differences in mechanisms of cardiotoxins, melittin and other myotoxins (1996) Toxicon, 34, pp. 1301-1311Florendo, J.A., Reger, J.F., Law, P.K., Electrophysiologic differences between mouse extensor digitorum lungus and soleus (1983) Exp. Neurol., 82, pp. 404-412Freitas, E.M.S., Dal Pai-Silva, M., Cruz-Höfling, M.A., Histoenzymological and ultrastructural changes in lateral muscle fibers of Oreochromis niloticus (Teleostei: Cichlidae) after local injection of veratrine (2001) Histochem. Cell Biol., 116, pp. 525-534Freitas, E.M.S., Dal Pai-Silva, M., Cruz-Höfling, M.A., Histochemical differences in the responses of predominantly fast-twitch glycolytic muscle and slow-twitch oxidative muscle to veratrine (2002) Toxicon, , in pressGarcia, M.C., Díaz, A.F., Godinez, R., Sánchez, J.A., Effect of sodium deprivation on contraction and charge movement in frog skeletal muscle fibers (1992) J. Muscle Res. Cell Motil., 13, pp. 354-365Honerjager, P., Cardioactive substances that prolong tlie open state of sodium channels (1982) Rev. Physiol. Biochem. Pharmacol., 92, pp. 1-74Honerjager, P., Dugas, M., Zong, X.-G., Mutually exclusive action of cationic veratridine and cevadine at an intracellular site of cardiac sodium channel (1992) J. Gen. 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Observation Of The Bone Matrix Structure Of Intact And Regenerative Zones Of Tibias By Atomic Force Microscopy

The structure of bone matrix of rat tibia from an intact and regenerative zones of tibias was studied using atomic force spectroscopy. The young male adult rats (wistar) were used for the study with corporal masses between 250 and 300 g. A 1.5-mm-diam hole was drilled in one cortical tibia surface to provoke the injury and the healing course was monitored. Atomic force spectroscopic images were used at different magnifications to identify the time dependence of the osteoblast activity, measured by the increase in the area of neoformed primary bone and in the organization of the collagen fibers.19410421045Manolagas, S.C., (1998) Aging-Clinical Exp. Res., 10, p. 182Rodan, G.A., Martin, T.J., (2000) Science, 289, p. 1508Silver, I.A., Murrills, R.J., Etherington, D.J., (1998) Exp. Cell Res., 175, p. 266Rodan, G.A., (1997) Bone (N.Y.), 20, p. 1Sandmaier, B.M., Sorb, R., Kniley, J., Hardy, W., Black, M., Moseley, A., Deans, R., (1998) Blood, 92, p. 473Michaelsson, K., Baron, J.A., Farahmand, B.Y., Johenll, O., Magnusson, C., Persson, P.G., Persson, I., Ljunghall, S., (1998) Br. Med. J., 316, p. 1858Bonadio, J., Smiley, E., Patil, P., Goldstein, S., (1999) Nature Med., 5, p. 753Freitas, I.G.F., Baranauskas, V., Cruz-Höfling, M.A., (2000) Appl. Surf. Sci., 154-155, p. 548Binnig, G., Quate, C.F., Gerber, Ch., (1986) Phys. Rev. Lett., 56, p. 930Yang, J., Shao, Z., (1995) Micron, 26, p. 35Wickramasinghe, H.R., (2000) Acta Mater., 48, p. 347Baranauskas, V., Vidal, B.C., Parizotto, N.A., (1998) Appl. Biochem. Biotech., 69, p. 91Ducy, P., Schinke, T., Karsenty, G., (2000) Science, 289, p. 1501Teitelbaum, S.L., (2000) Science, 289, p. 150

Electrophysiological And Ultrastructural Analysis Of The Neuromuscular Blockade And Miotoxicity Induced By The Micrurus Nigrocinctus Snake Venom.

Micrurus nigrocinctus is the most abundant coral snake in Central America. The venom of this specie induced a concentration-dependent (10-20 micrograms/ml) depolarization in the isolated mouse phrenic nerve-diaphragm preparations incubated at 37 degrees C. d-Tubocurarine (10 micrograms/ml) and (alpha beta ungarotoxin (3-5 micrograms/ml) were able to partially protect against the depolarization induced by the venom (10 micrograms/ml), suggesting the involvement of subsynaptic cholinergic receptors. This venom (10 micrograms/ml) also increased the frequency and amplitude of miniature end-plate potentials (mepps) during the first 10-20 min of incubation. Subsequently, the mepps progressively decreased and disappeared after 60 min. These responses were accompanied by ultrastructural changes involving the nerve terminals, the subsynaptic junctional folds and the muscle mitochondria. The synaptic gutter was shallow and, very often, "shrunken" terminals with omega-shaped axolemmal indentations and a decreased number of synaptic vesicles were present. A common finding was the presence of numerous finger-like, membrane-bounded bodies interposed between the terminal and the Schwann cells or postsynaptic sarcolemma. The preincubation of the venom with specific antivenom or the incubation of the preparations at room temperature (24-26 degrees C) reduced the number and intensity of the ultrastructural alterations. The last finding suggests the involvement of a enzymatic process, probably a phospholipase A2, present in the venom. There was a good correlation between the electrophysiological and ultrastructural effects induced by the venom which allow us to conclude that M. nigrocinctus venom has a presynaptic action in the initial stages of intoxication followed by sub- and postsynaptic effects, the last being the most important cause of neuromuscular blockade. A direct action of the venom on muscle fibers may also contributes to the irreversible blockade.49429029

Low-level Laser Therapy Promotes Vascular Endothelial Growth Factor Receptor-1 Expression In Endothelial And Nonendothelial Cells Of Mice Gastrocnemius Exposed To Snake Venom

Crotalinae snake venoms cause severe local myonecrosis and microvasculature failure at the bite site. We evaluated whether low-level laser therapy (LLLT) could accelerate angiogenesis and myoregeneration in male Swiss mice injected with Bothrops moojeni venom through immunohistochemistry of the vascular endothelial growth factor receptor-1 (VEGFR-1). Envenomed gastrocnemius was either unirradiated (V) or irradiated with HeNe (VHN, 632.8 nm) or GaAs (VGA, 904 nm, 10000 Hz). Animals sacrificed at 3 and 12 h were irradiated once (4 J cm-2), at 24 h (twice) and at 3, 7, 21 days (4, 8, 22 times, respectively). At 3 days, LLLT increased angiogenesis (80%:HeNe vs 40%:GaAs), decreased neutrophils and increased proliferation of regenerating cells. However, after 21 days, myoregeneration observed in the VHN group appeared delayed compared with the V group. As LLLT improved revascularization, the suggestive delay in myoregeneration could be a dose-response inhibitory effect caused by multiple irradiations in myogenesis. The immunodetection of VEGFR-1 in neutrophils, macrophages, satellite cells, fibroblasts, Schwann cells and skeletal and smooth muscle fibers (not seen in saline-controls) at only the acute stages of envenoming suggests a mediator role for VEGFR-1 in local alterations. This is the first time that VEGFR-1 expression, and its modulation by photostimulation, has been demonstrated in endothelial and nonendothelial cells of snake envenomed skeletal muscle. © 2010 The American Society of Photobiology.872418426Da Silva, C.J., Jorge, M.T., Ribeiro, L.A., Epidemiology of snakebite in a central region of Brazil (2003) Toxicon, 41 (2), pp. 251-255. , DOI 10.1016/S0041-0101(02)00287-8, PII S0041010102002878Bieber, A.L., Metal and non-protein constituents in snake venoms (1979) Handbook of Experimental Pharmacology, pp. 295-306. , In (Edited by C. Y. Lee), pp. Springer-Verlag, New YorkLeite, L.C.C., Furtado, M.F.D., Correa, T.C., Raw, I., Characterization of the snake venoms from seven Brazilian species of Bothrops by FPLC anion-exchange chromatography (1992) Comp. Biochem. Physiol., 102, pp. 515-520Ownby, C.L., Locally acting agents: Myotoxins, hemorrhagic toxins and dermonecrotic factors (1990) Handbook of Toxinology, pp. 602-654. , In (Edited by W. T. 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