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Lopap, found in the bristles of Lonomia obliqua caterpillar, is the first exogenous prothrombin activator that shows serine protease-like activity, independent of prothrombinase components and unique lipocalin reported to interfere with hemostasis mechanisms. To assess the action of an exogenous prothrombin activator reversing the anticoagulant and antihemostatic effect induced by low molecular weight heparin (LMWH), male New Zealand rabbits (N = 20, weighing 3.8-4.0 kg) allocated to 4 groups were anticoagulated with 1800 IU/kg LMWH (iv) over 2 min, followed by iv administration of saline (SG) or recombinant Lopap (rLopap) at 1 µg/kg (LG1) or 10 µg/kg (LG10), 10 min after the injection of LMWH, in a blind manner. Control animals (CG) were treated only with saline. The action of rLopap was assessed in terms of activated partial thromboplastin time (aPTT), prothrombin fragment F1+2, fibrinogen, and ear puncture bleeding time (BT) at 5, 10, 15, 17, 20, 30, 40, 60, and 90 min after initiation of LMWH infusion. LG10 animals showed a decrease of aPTT in more than 50% and BT near to normal baseline. The level of prothrombin fragment F1+2 measured by ELISA had a 6-fold increase with rLopap treatment (10 µg/kg) and was inversely proportional to BT in LMWH-treated animals. Thus, Lopap, obtained in recombinant form using E. coli expression system, was useful in antagonizing the effect of LMWH through direct prothrombin activation, which can be a possible strategy for the reversal of bleeding and anticoagulant events.

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Lopap, found in the bristles of Lonomia obliqua caterpillar, is the first exogenous prothrombin activator that shows serine protease-like activity, independent of prothrombinase components and unique lipocalin reported to interfere with hemostasis mechanisms. To assess the action of an exogenous prothrombin activator reversing the anticoagulant and antihemostatic effect induced by low molecular weight heparin (LMWH), male New Zealand rabbits (N = 20, weighing 3.8-4.0 kg) allocated to 4 groups were anticoagulated with 1800 IU/kg LMWH (iv) over 2 min, followed by iv administration of saline (SG) or recombinant Lopap (rLopap) at 1 µg/kg (LG1) or 10 µg/kg (LG10), 10 min after the injection of LMWH, in a blind manner. Control animals (CG) were treated only with saline. The action of rLopap was assessed in terms of activated partial thromboplastin time (aPTT), prothrombin fragment F1+2, fibrinogen, and ear puncture bleeding time (BT) at 5, 10, 15, 17, 20, 30, 40, 60, and 90 min after initiation of LMWH infusion. LG10 animals showed a decrease of aPTT in more than 50% and BT near to normal baseline. The level of prothrombin fragment F1+2 measured by ELISA had a 6-fold increase with rLopap treatment (10 µg/kg) and was inversely proportional to BT in LMWH-treated animals. Thus, Lopap, obtained in recombinant form using E. coli expression system, was useful in antagonizing the effect of LMWH through direct prothrombin activation, which can be a possible strategy for the reversal of bleeding and anticoagulant events.

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AIM: Vein reconstruction using grafts may prevent sequelae of venous interruption or lesion. Autologous vein is sometimes unsuitable or absent for a vascular restoration. The aim of this study was to study glutaraldehyde-treated homologous vein graft as vein substitute and compare it with autologous vein as a substitute for a vena cava segment in rabbits. METHODS: Sixty rabbits were allocated into two groups: autologous vein graft (AG), and glutaraldehyde-treated homologous vein graft (HG). Each group was subdivided into three subgroups (N.=10) to be studied at: 24 hours, 14 days, and 28 days. The veins were treated in 0.19% glutaraldehyde, pH=7.4, for 1 hour and kept at 4 degrees C in saline with added gentamicin and amphotericin B. The animals received benzanthine penicillin on the day of graft implantation and heparin only during surgery. The grafts were implanted into the vena cava. Anastomosis was performed with interrupted sutures. Cavography was performed, after surgery, and at the time the animals were killed. Evaluation of the veins was made macroscopically and by light and scanning electron microscopy. RESULTS: Fibrosis was seen around the grafts at 14 and 28 days, with no difference in intensity between the groups. Cavography performed before euthanasia of the animals showed 4 partial thrombi in AG (2 at 24 hours and 2 at 14 days), 3 in HG (2 at 24 hours and 1 on day 14), and 4 occlusive thombi in HG (3 at 14 days and 1 at 28 days). Macroscopic examination did not show any thrombus in AG. In HG, two partial thrombi were confirmed at 24 hours and three occlusive thrombi at 14 days. There was no statistical difference in relation to patency between the two groups. At 14 and 28 days, the histological sections showed intimal hyperplasia of similar intensity and variable distribution in both groups. Evaluation by electron microscopy showed at 24 hours lesion areas characterized by absence of the endothelium on the graft surface, presence of inflammatory cells, and, at some sites, presence of mural thrombi in AG and HG. Both groups at 14 and 28 days showed endothelial cells covering the lesion area on the graft surface, this covering being larger in AG than in HG. CONCLUSIONS: In the studied model, both grafts behaved similarly in relation to patency and morphological characteristics. This suggests that the glutaraldehyde-treated graft can be a promising alternative for vein reconstruction, justifying further animal studies with the aim of using it in human surgery.

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AIM: This study aimed at assessing the accuracy of ultrasound (US) in the diagnosis of recent deep vein thrombosis (DVT) in an experimental study in dogs. DESIGN: blinded and randomized experimental study. Twenty dogs were randomly divided in two groups: control group (CG) and thrombosis group (TG). US was performed in the pre- and postoperative period. Phlebography was performed immediately prior to the postoperative US. After the second US, a surgery was performed to detect whether thrombus was present or not. US results were compared to those of phlebography and surgical findings. RESULTS: In all dogs, inferior vena cava (IVC) was compressible. The relations of IVC diameter with the aorta were higher (P<0.005) in TG than in CG. Spectral Doppler in spontaneous breathing, tissue harmonic imaging, power Doppler and B flow showed sensitivity, specificity and accuracy of 1. Phlebography presented sensitivity of 90%, specificity of 80% and accuracy of 85%, when compared to surgical finding. CONCLUSIONS: For the diagnosis of recent DVT in the experimental model used, venous compressibility proved to be inefficient. The ratio of IVC diameter to aorta, when increased, suggests thrombosis. The use of new US technological advances increases accuracy. Phlebography was less accurate than US.

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The venom of Lonomia obliqua caterpillar may induce a hemorrhagic syndrome in humans, and blood incoagulability by afibrinogenemia when intravenously injected in laboratory animals. The possible antithrombotic and thrombolytic activities of L. obliqua caterpillar bristle extract (LOCBE) were evaluated in this study. The minimal intravenous dose of the extract necessary to induce afibrinogenemia and anticoagulation was 3.0 and 10.0 microg protein/kg body weight for rabbits and rats, respectively. In rabbits, this dose induced total blood incoagulability for at least 10 h and did not reduce the weight of preformed venous thrombi, in contrast to streptokinase (30,000 IU/kg). In rats, pretreatment with 5.0 and 10.0 microg/kg LOCBE prevented the formation of thrombi induced by venous stasis or by injury to the venous endothelium. The dose of 5.0 microg/kg LOCBE did not modify blood coagulation assay parameters but increased bleeding time and decreased plasma factor XIII concentration. When the extract was administered to rats at the dose of 10.0 microg/kg, the blood was totally incoagulable for 6 h. These data show that LOCBE was effective in preventing experimental venous thrombosis in rats, justifying further studies using purified fractions of the extract to clarify the mechanisms of this effect.

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The venom of Lonomia obliqua caterpillar may induce a hemorrhagic syndrome in humans, and blood incoagulability by afibrinogenemia when intravenously injected in laboratory animals. The possible antithrombotic and thrombolytic activities of L. obliqua caterpillar bristle extract (LOCBE) were evaluated in this study. The minimal intravenous dose of the extract necessary to induce afibrinogenemia and anticoagulation was 3.0 and 10.0 æg protein/kg body weight for rabbits and rats, respectively. In rabbits, this dose induced total blood incoagulability for at least 10 h and did not reduce the weight of preformed venous thrombi, in contrast to streptokinase (30,000 IU/kg). In rats, pretreatment with 5.0 and 10.0 æg/kg LOCBE prevented the formation of thrombi induced by venous stasis or by injury to the venous endothelium. The dose of 5.0 æg/kg LOCBE did not modify blood coagulation assay parameters but increased bleeding time and decreased plasma factor XIII concentration. When the extract was administered to rats at the dose of 10.0 æg/kg, the blood was totally incoagulable for 6 h. These data show that LOCBE was effective in preventing experimental venous thrombosis in rats, justifying further studies using purified fractions of the extract to clarify the mechanisms of this effect

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Com base na utilização empírica do metronidazol no tratamento de picada de cobra e em uma possível ação fibrinolítica, alguns médicos brasileiros o usaram no tratamento de trombose venosa profunda, embora não existam estudos que justifiquem tal emprego. OBJETIVO: Verificar possível existência de efeito antitrombótico do metronidazol em modelo de trombose induzida na veia cava de ratos e uma eventual ação anticoagulante ou fibrinolítica dessa droga. MÉTODO: 71 ratos da raça Wistar foram usados em 4 experimentos. No primeiro, 29 animais foram divididos em três grupos: controle (solução salina), heparina (200UI/kg) e metronidazol (6mg/kg). 10 minutos após a injeção IV a trombose foi induzida pela ligadura da veia cava, distalmente à veia renal esquerda. Três horas depois, a veia cava era removida e aberta, e o trombo, se presente, retirado para pesagem, sendo o sangue do animal retirado para medida do TTPA, TT e TP. No segundo experimento, o sangue foi retirado dos animais (n=18) 10 minutos após a injeção das drogas, para medida dos mesmos parâmetros. No terceiro experimento os animais (n=22) foram injetados com metronidazol ou salina e o tempo de lise de euglobulina medido 3 horas após. No quarto experimento, 22 animais foram sorteados para um de dois grupos: controle, com injeção IV de 12ml de salina ou metronidazol, (12mg/kg), sendo a trombose induzida 10 minutos após. RESULTADOS: Não houve diferença entre os grupos controles e os grupod tratados com metronidazol, em relação à incidência e peso dos trombos e com relação aos parâmetros hemostáticos estudados. Nos animais tratados com heparina não houve formção de trombos, havendo aumento de TTPA e de TT aos 10 minutos, e de TT, após 3 horas. CONCLUSÄO: No modelo experimental empregado, o metronidazol não mostrou efeito antitrombótico e também ação sobre o parâmetros hemostáticos estudados, sugerindo que o uso clínico dessa droga no tratamento de trombose venosa é duvidoso, a menos que outros estudos venham demosntrar algum efeito benéfico.

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The present study evaluates platelet activation following application of an Esmarch bandage and a tourniquet, procedures commonly employed to provide a bloodless operative field during limb surgery. Platelet aggregation was increased in blood samples taken from rabbits 60 min after an Esmarch bandage was applied to one thigh and immediately released. When this treatment was combined with the application of a tourniquet for 60 min, a procedure which alone did not affect platelet aggregation, results were similar to those obtained following the Esmarch bandage alone. These data suggest that tissue compression produced by application of an Esmarch bandage, but not the ischemia derived from the tourniquet, produced platelet aggregation

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The action of threem different topical heparinoids on the evolution of experimental thrombophlebitis was studies. Thrombophlebitis was induced in the marginal vein of the ear of rabbits by stasis and inection of hypertonic glucose solution. Forty-eight hours later animals were allocated to three treatment groups and a control group. The substances were applied over the affected vein three times a day for 6 days and the ears inspected daily by transillumination. After 7 days, the animals were killed and anatomopathological studies performed. No difference in thrombus frequency or inflamatory reaction was observed between the animals treated with heparinoids and the control group, or among the treated groups

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