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  1. Article ; Online: Platelet participation in the pathogenesis of dermonecrosis induced by Loxosceles gaucho venom.

    Tavares, F L / Peichoto, M E / Marcelino, J R / Barbaro, K C / Cirillo, M C / Santoro, M L / Sano-Martins, I S

    Human & experimental toxicology

    2016  Volume 35, Issue 6, Page(s) 666–676

    Abstract: Loxosceles gaucho spider venom induces in vitro platelet activation and marked thrombocytopenia in rabbits. Herein, we investigated the involvement of platelets in the development of the dermonecrosis induced by L. gaucho venom, using thrombocytopenic ... ...

    Abstract Loxosceles gaucho spider venom induces in vitro platelet activation and marked thrombocytopenia in rabbits. Herein, we investigated the involvement of platelets in the development of the dermonecrosis induced by L. gaucho venom, using thrombocytopenic rabbits as a model. L. gaucho venom evoked a drop in platelet and neutrophil counts 4 h after venom injection. Ecchymotic areas at the site of venom inoculation were noticed as soon as 4 h in thrombocytopenic animals but not in animals with initial normal platelet counts. After 5 days, areas of scars in thrombocytopenic animals were also larger, evidencing the marked development of lesions in the condition of thrombocytopenia. Histologically, local hemorrhage, collagen fiber disorganization, and edema were more severe in thrombocytopenic animals. Leukocyte infiltration, predominantly due to polymorphonuclears, was observed in the presence or not of thrombocytopenia. Thrombus formation was demonstrated by immunohistochemistry at the microvasculature, and it occurred even under marked thrombocytopenia. Taken together, platelets have an important role in minimizing not only the hemorrhagic phenomena but also the inflammatory and wound-healing processes, suggesting that cutaneous loxoscelism may be aggravated under thrombocytopenic conditions.
    MeSH term(s) Animals ; Blood Cell Count ; Blood Platelets/physiology ; Disease Models, Animal ; Endothelium, Vascular/drug effects ; Endothelium, Vascular/metabolism ; Necrosis ; Neutrophils/drug effects ; Phagocytosis/drug effects ; Phosphoric Diester Hydrolases/toxicity ; Prothrombin Time ; Rabbits ; Skin/blood supply ; Skin/drug effects ; Skin/pathology ; Skin Diseases/blood ; Skin Diseases/chemically induced ; Skin Diseases/pathology ; Spider Venoms/toxicity ; Thrombocytopenia/blood ; von Willebrand Factor/analysis
    Chemical Substances Spider Venoms ; loxosceles venom ; von Willebrand Factor ; Phosphoric Diester Hydrolases (EC 3.1.4.-)
    Language English
    Publishing date 2016-06
    Publishing country England
    Document type Journal Article
    ZDB-ID 1027454-6
    ISSN 1477-0903 ; 0144-5952 ; 0960-3271
    ISSN (online) 1477-0903
    ISSN 0144-5952 ; 0960-3271
    DOI 10.1177/0960327115597983
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  2. Article ; Online: A functional and thromboelastometric-based micromethod for assessing crotoxin anticoagulant activity and antiserum relative potency against Crotalus durissus terrificus venom.

    Prezoto, B C / Tanaka-Azevedo, A M / Marcelino, J R / Tashima, A K / Nishiduka, E S / Kapronezai, J / Mota, J O / Rocha, M M T / Serino-Silva, C / Oguiura, N

    Toxicon : official journal of the International Society on Toxinology

    2018  Volume 148, Page(s) 26–32

    Abstract: The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies on traditional rodent in vivo lethality assay. ... ...

    Abstract The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies on traditional rodent in vivo lethality assay. ED
    MeSH term(s) Animals ; Antivenins/pharmacology ; Blood Coagulation/drug effects ; Chickens ; Crotalid Venoms/immunology ; Crotalid Venoms/toxicity ; Crotalus ; Crotoxin/toxicity ; Horses ; Neutralization Tests ; Thrombelastography/methods
    Chemical Substances Antivenins ; Crotalid Venoms ; Crotoxin (9007-40-3)
    Language English
    Publishing date 2018-04-11
    Publishing country England
    Document type Journal Article
    ZDB-ID 204479-1
    ISSN 1879-3150 ; 0041-0101
    ISSN (online) 1879-3150
    ISSN 0041-0101
    DOI 10.1016/j.toxicon.2018.04.009
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  3. Article ; Online: An alternative micromethod to access the procoagulant activity of Bothrops jararaca venom and the efficacy of antivenom.

    Oguiura, N / Kapronezai, J / Ribeiro, T / Rocha, M M T / Medeiros, C R / Marcelino, J R / Prezoto, B C

    Toxicon : official journal of the International Society on Toxinology

    2014  Volume 90, Page(s) 148–154

    Abstract: The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies largely on traditional rodent lethality assay (LD50). However, adequately validated in vitro tests should be introduced for assessing antivenom ... ...

    Abstract The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies largely on traditional rodent lethality assay (LD50). However, adequately validated in vitro tests should be introduced for assessing antivenom neutralizing capacity in plasma of immunized horses as well as for in-process quality control. The dynamic of fibrin formation in recalcified avian plasma samples is extremely slow, when compared to that presented by mammalian plasmas. In this study, we present one new coagulant assay, by performing dose-response curve after plotting the clotting time (CT) parameter of the ROTEM profile of recalcified chicken plasma samples (target) against semi-logarithmic doses of Bothrops jararaca venom (agonist), either in absence or in presence of the semi-logarithmic doses of anti-bothropic serum (ABS) (antagonist). The mean coagulant dose 50% (CD50) was defined as the quantity of venom (in μg) which reduces CT to 900 s, between minimum and maximum responses. The CT induced by 5CD50 of the venom was used as the control for calculating the effective dose (ED) of each batch of ABS. ED was defined as the ABS dose (nanoliters, nL) at which CT induced by one amount of venom corresponding to 5CD50 is displaced to the maximum threshold (1800 s). Five batches of the ABS, previously assayed for their lethality neutralizing activity (ED50) were assayed. The correlation coefficient (r) between both in vitro (ED) and in vivo (ED50) values was 0.87 (p value < 0.05). We propose this micro method as highly sensitive for characterization and quantification of possible procoagulant activity of small doses of snake venoms (nanograms) and for detecting small doses (nanoliters) of specific antibodies against this effect in little volume samples of biological fluids.
    MeSH term(s) Animals ; Antivenins/pharmacology ; Bothrops ; Chickens ; Coagulants/toxicity ; Crotalid Venoms/toxicity ; Female ; Male ; Prothrombin Time
    Chemical Substances Antivenins ; Coagulants ; Crotalid Venoms
    Language English
    Publishing date 2014-11
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 204479-1
    ISSN 1879-3150 ; 0041-0101
    ISSN (online) 1879-3150
    ISSN 0041-0101
    DOI 10.1016/j.toxicon.2014.08.004
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  4. Article: Characterization of anti-crotalic antibodies

    Guidolin, F.R / Tambourgi, D.V / Guidolin, R / Marcelino, J.R / Okamoto, C.K / Magnoli, F.C / Queiroz, G.P / Dias da Silva, W

    Toxicon. 2013 May, v. 66

    2013  

    Abstract: Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated ... ...

    Abstract Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated components, crotapotin and phospholipase A2 (PLA2). Treatment of the victims with specific antiserum is the unique effective therapeutic measure. The ability of anti-Crotalus antisera produced by the routine using crude venom to immunize horses or purified crotoxin and PLA2 as individual immunogens was compared. Antisera obtained from horses immunized with C. durissus terrificus crude venom were able to recognize and neutralize not only the toxins presents in C. durissus terrificus, but also the ones present in the venoms from C. d. collilineatus, C. d. cascavella and C. d. marajoensis. Antisera from horses immunized with individual crotoxin or PLA2, although in lesser titers, were also able of recognizing the toxins in all four Crotalus species and neutralize the lethality of the C. d. terrificus venom.
    Keywords Crotalus durissus terrificus ; antibodies ; antigens ; antiserum ; horses ; neurotoxins ; phospholipase A2 ; snake bites ; snakes ; venoms ; Brazil
    Language English
    Dates of publication 2013-05
    Size p. 7-17.
    Publishing place Elsevier Ltd
    Document type Article
    ZDB-ID 204479-1
    ISSN 1879-3150 ; 0041-0101
    ISSN (online) 1879-3150
    ISSN 0041-0101
    DOI 10.1016/j.toxicon.2013.01.015
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  5. Article ; Online: Characterization of anti-crotalic antibodies.

    Guidolin, F R / Tambourgi, D V / Guidolin, R / Marcelino, J R / Okamoto, C K / Magnoli, F C / Queiroz, G P / Dias da Silva, W

    Toxicon : official journal of the International Society on Toxinology

    2013  Volume 66, Page(s) 7–17

    Abstract: Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated ... ...

    Abstract Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated components, crotapotin and phospholipase A2 (PLA2). Treatment of the victims with specific antiserum is the unique effective therapeutic measure. The ability of anti-Crotalus antisera produced by the routine using crude venom to immunize horses or purified crotoxin and PLA2 as individual immunogens was compared. Antisera obtained from horses immunized with C. durissus terrificus crude venom were able to recognize and neutralize not only the toxins presents in C. durissus terrificus, but also the ones present in the venoms from C. d. collilineatus, C. d. cascavella and C. d. marajoensis. Antisera from horses immunized with individual crotoxin or PLA2, although in lesser titers, were also able of recognizing the toxins in all four Crotalus species and neutralize the lethality of the C. d. terrificus venom.
    MeSH term(s) Animals ; Antivenins/biosynthesis ; Antivenins/immunology ; Antivenins/pharmacology ; Biological Assay ; Crotalid Venoms/immunology ; Crotalid Venoms/toxicity ; Crotoxin/immunology ; Disease Models, Animal ; Horses/immunology ; Lethal Dose 50 ; Male ; Mice ; Neurotoxins/immunology ; Neutralization Tests ; Phospholipases A2/immunology ; Snake Bites/drug therapy ; Snake Bites/immunology ; Snake Bites/mortality ; Survival Analysis
    Chemical Substances Antivenins ; Crotalid Venoms ; Neurotoxins ; Crotoxin (9007-40-3) ; Phospholipases A2 (EC 3.1.1.4)
    Language English
    Publishing date 2013-05
    Publishing country England
    Document type Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 204479-1
    ISSN 1879-3150 ; 0041-0101
    ISSN (online) 1879-3150
    ISSN 0041-0101
    DOI 10.1016/j.toxicon.2013.01.015
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  6. Article: Effect of beta-propiolactone treatment on the complement activation mediated by equine antisera.

    Guidolin, R / Morais, J F / Stephano, M A / Marcelino, J R / Yamaguchi, I K / Higashi, H G

    Revista do Instituto de Medicina Tropical de Sao Paulo

    1997  Volume 39, Issue 2, Page(s) 119–122

    Abstract: Reduction of complement activation through an alteration of the Fc fragment of immunoglobulins by beta-propiolactone treatment was carried out in equine antisera raised against rabies virus, Bothrops venoms and diphtherial toxin. Results were evaluated ... ...

    Abstract Reduction of complement activation through an alteration of the Fc fragment of immunoglobulins by beta-propiolactone treatment was carried out in equine antisera raised against rabies virus, Bothrops venoms and diphtherial toxin. Results were evaluated by means of an anaphylactic test performed on guinea-pigs, and compared to the ones obtained with the same sera purified by saline precipitation (ammonium sulfate), followed or not by enzymatic digestion with pepsin. Protein purity levels for antibothropic serum were 184.5 mg/g and 488.5 mg/g in beta-propiolactone treated and pepsin-digested sera, respectively. The recovery of specific activity was 100% and 62.5% when using antibothropic serum treated by beta-propiolactone and pepsin digestion, respectively. The antidiphtherial and anti-rabies sera treated with beta-propiolactone and pepsin presented protein purity levels of 5,698 and 7,179 Lf/g, 16,233 and 6,784 IU/g, respectively. The recovery of specific activity for these antisera were 88.8%, 77.7%, 100% and 36.5%, respectively. beta-propiolactone treatment induced a reduction in complement activation, tested "in vivo", without significant loss of biological activity. This treatment can be used in the preparation of heterologous immunoglobulins for human use.
    MeSH term(s) Animals ; Anti-Infective Agents, Local/pharmacology ; Complement Activation/drug effects ; Complement Activation/immunology ; Guinea Pigs ; Horses ; Immune Sera/immunology ; Propiolactone/pharmacology
    Chemical Substances Anti-Infective Agents, Local ; Immune Sera ; Propiolactone (6RC3ZT4HB0)
    Language English
    Publishing date 1997-03
    Publishing country Brazil
    Document type Journal Article
    ZDB-ID 128928-7
    ISSN 1678-9946 ; 0036-4665
    ISSN (online) 1678-9946
    ISSN 0036-4665
    DOI 10.1590/s0036-46651997000200010
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  7. Article: Neutralization of crotaline snake venoms from Central and South America by antivenoms produced in Brazil and Costa Rica.

    Bogarín, G / Morais, J F / Yamaguchi, I K / Stephano, M A / Marcelino, J R / Nishikawa, A K / Guidolin, R / Rojas, G / Higashi, H G / Gutiérrez, J M

    Toxicon : official journal of the International Society on Toxinology

    2000  Volume 38, Issue 10, Page(s) 1429–1441

    Abstract: A study was performed on the ability of antivenoms, produced in Brazil and Costa Rica, to neutralize lethal, hemorrhagic and coagulant activities of the venoms of 16 species of Central and South American snakes of the subfamily Crotalinae. Neutralization ...

    Abstract A study was performed on the ability of antivenoms, produced in Brazil and Costa Rica, to neutralize lethal, hemorrhagic and coagulant activities of the venoms of 16 species of Central and South American snakes of the subfamily Crotalinae. Neutralization of lethality was studied by two different methods routinely used in the quality control of antivenoms at Instituto Butantan (IB) and Instituto Clodomiro Picado (ICP). Both antivenoms neutralized the majority of the venoms studied, but the values of effective doses 50% (ED(50)) differed markedly depending on the method used. In general, higher potencies were obtained with the method of ICP, where a challenge dose corresponding to 4 LD(50)s is used, than with the method of IB, where a challenge dose of 5 LD(50)s is employed. All venoms induced hemorrhagic activity in the mouse skin test, which was effectively neutralized by the two antivenoms. All venoms, except those of Porthidium nasutum and Bothriechis lateralis, induced coagulation of human plasma in vitro and both antivenoms were effective in the neutralization of this activity. In conclusion, our results provide evidence of an extensive cross reactivity between these antivenoms and Central and South American crotaline snake venoms.
    MeSH term(s) Animals ; Antivenins/administration & dosage ; Antivenins/pharmacology ; Blood Coagulation/drug effects ; Blood Coagulation/physiology ; Blood Coagulation Tests ; Brazil ; Coagulants/antagonists & inhibitors ; Costa Rica ; Cross Reactions ; Crotalid Venoms/antagonists & inhibitors ; Crotalid Venoms/toxicity ; Hemolysis/drug effects ; Hemorrhage/prevention & control ; In Vitro Techniques ; Injections, Intraperitoneal ; Lethal Dose 50 ; Mice ; Neutralization Tests
    Chemical Substances Antivenins ; Coagulants ; Crotalid Venoms
    Language English
    Publishing date 2000-10
    Publishing country England
    Document type Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 204479-1
    ISSN 1879-3150 ; 0041-0101
    ISSN (online) 1879-3150
    ISSN 0041-0101
    DOI 10.1016/s0041-0101(99)00236-6
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  8. Article: Antigenic cross-reactivity among components of Brazilian Elapidae snake venoms.

    Higashi, H G / Guidolin, R / Caricati, C P / Fernandes, I / Marcelino, J R / Morais, J F / Yamagushi, I K / Stephano, M A / Dias-da-Silva, W / Takehara, H A

    Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas

    1995  Volume 28, Issue 7, Page(s) 767–771

    Abstract: Snake venoms from M. corallinus (LD50 = 7.1 +/- 0.83 micrograms), M. frontalis (LD50 = 19.3 +/- 3.13 micrograms), M. ibiboboca (LD50 = 19.8 +/- 2.07 micrograms) and M. spiixi (LD50 = 6.7 +/- 1.25 micrograms) (family Elapidae, genus Micrurus) injected ... ...

    Abstract Snake venoms from M. corallinus (LD50 = 7.1 +/- 0.83 micrograms), M. frontalis (LD50 = 19.3 +/- 3.13 micrograms), M. ibiboboca (LD50 = 19.8 +/- 2.07 micrograms) and M. spiixi (LD50 = 6.7 +/- 1.25 micrograms) (family Elapidae, genus Micrurus) injected into horses alone or in combination (M. corallinus with M. frontalis) elicit antibody production, as indicated in vivo by neutralization of venom lethality and in vitro by enzyme-linked immunosorbent assay (ELISA), immunoelectrophoresis (IE) and Western blotting (WB). Venom lethality was efficiently neutralized by the antisera, with the monovalent antivenoms being more efficient than the bivalent antivenom. Antibodies against venom components were detected by all antisera at different titers by ELISA. Upon IE, antisera against M. spiixi and M. frontalis venoms cross-reacted with the four types of venoms studied and recognized several molecular components, the precipitin lines obtained had distinct intensities and electrophoretic motilities, whereas the antivenom against M. corallinus only recognized components of its venom but not of the others. All antivenoms cross-reacted with all the elapid venoms in WB revealing several bands with distinct MWs in M. corallinus and M. spiixi venoms, two very sharp and separate bands in M. corallinus venom and a very sharp band of high MW together with several other smaller and faint bands in M. frontalis venom. The data indicate that snake venoms of the genus Micrurus are good immunogens that contain many cross-reactive molecules, and that their toxic components are neutralized more effectively by monovalent rather than by bivalent antivenom.
    MeSH term(s) Animals ; Antivenins/biosynthesis ; Brazil ; Cross Reactions ; Elapid Venoms/immunology ; Horses ; Lethal Dose 50
    Chemical Substances Antivenins ; Elapid Venoms
    Language English
    Publishing date 1995-07
    Publishing country Brazil
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 786234-9
    ISSN 0100-879X
    ISSN 0100-879X
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  9. Article: Characterization of anti-crotalic antibodies

    Guidolin, F.R. / Tambourgi, D.V. / Guidolin, R. / Marcelino, J.R. / Okamoto, C.K. / Magnoli, F.C. / Queiroz, G.P. / Dias da Silva, W.

    Toxicon

    Volume v. 66

    Abstract: Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated ... ...

    Abstract Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated components, crotapotin and phospholipase A2 (PLA2). Treatment of the victims with specific antiserum is the unique effective therapeutic measure. The ability of anti-Crotalus antisera produced by the routine using crude venom to immunize horses or purified crotoxin and PLA2 as individual immunogens was compared. Antisera obtained from horses immunized with C. durissus terrificus crude venom were able to recognize and neutralize not only the toxins presents in C. durissus terrificus, but also the ones present in the venoms from C. d. collilineatus, C. d. cascavella and C. d. marajoensis. Antisera from horses immunized with individual crotoxin or PLA2, although in lesser titers, were also able of recognizing the toxins in all four Crotalus species and neutralize the lethality of the C. d. terrificus venom.
    Keywords neurotoxins ; horses ; antibodies ; venoms ; Crotalus durissus terrificus ; phospholipase A2 ; snake bites ; snakes ; antiserum ; antigens
    Language English
    Document type Article
    ISSN 0041-0101
    Database AGRIS - International Information System for the Agricultural Sciences and Technology

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