Molecular and functional characterization of a new non-hemorrhagic metalloprotease from Bothrops jararacussu snake venom with antiplatelet activity

BjussuMP-II is an acidic low molecular weight metalloprotease (Mr similar to 24,000 and pI similar to 6.5), isolated from Bothrops jararacussu snake venom. The chromatographic profile in RP-HPLC and its N-terminal sequence confirmed its high purity level. Its complete cDNA was obtained by RT-PCR and the 615 bp codified for a mature protein of 205 amino acid residues. The multiple alignment of its deduced amino acid sequence and those of other snake venom metalloproteases showed a high structural similarity, mainly among class P-I proteases. The molecular modeling analysis of BjussuMP-II showed also conserved structural features with other SVMPs. BjussuMP-II did not induce hemorrhage, myotoxicity and lethality, but displayed dose-dependent proteolytic activity on fibrinogen, collagen, fibrin, casein and gelatin, keeping stable at different pHs, temperatures and presence of several divalent ions. BjussuMP-II did not show any clotting or anticoagulant activity on human citrated plasma, in contrast to its inhibitory effects on platelet aggregation. The aspects broached, in this work, provide data on the relationship between structure and function, in order to better understand the effects elicited by snake venom metalloproteases. (c) 2007 Elsevier B.V. All rights reserved.

Anti-hemorrhagic Activity of Four Brazilian Vegetable Species Against Bothrops jararaca Venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Gait training with partial body weight support during overground walking for individuals with chronic stroke: a pilot study

Background: It is not yet established if the use of body weight support (BWS) systems for gait training is effective per se or if it is the combination of BWS and treadmill that improves the locomotion of individuals with gait impairment. This study investigated the effects of gait training on ground level with partial BWS in individuals with stroke during overground walking with no BWS.Methods: Twelve individuals with chronic stroke (53.17 +/- 7.52 years old) participated of a gait training program with BWS during overground walking, and were evaluated before and after the gait training period. In both evaluations, individuals were videotaped walking at a self-selected comfortable speed with no BWS. Measurements were obtained for mean walking speed, step length, stride length and speed, toe-clearance, durations of total double stance and single-limb support, and minimum and maximum foot, shank, thigh, and trunk segmental angles.Results: After gait training, individuals walked faster, with symmetrical steps, longer and faster strides, and increased toe-clearance. Also, they displayed increased rotation of foot, shank, thigh, and trunk segmental angles on both sides of the body. However, the duration of single-limb support remained asymmetrical between each side of the body after gait training.Conclusions: Gait training individuals with chronic stroke with BWS during overground walking improved walking in terms of temporal-spatial parameters and segmental angles. This training strategy might be adopted as a safe, specific and promising strategy for gait rehabilitation after stroke.

Structural studies of BmooMP alpha-I, a non-hemorrhagic metalloproteinase from Bothrops moojeni venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The Venomics of Bothrops alternatus is a Pool of Acidic Proteins with Predominant Hemorrhagic and Coagulopathic Activities

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Coronariografia via transradial: curva de aprendizagem, avaliada por estudo multicêntrico

INTRODUÇÃO: A via de acesso femoral tem sido a preferida para os procedimentos diagnósticos e terapêuticos coronarianos, mas apresenta limitações, principalmente relacionadas a complicações vasculares e hemorrágicas. O aces-so transradial é uma técnica mais recentemente empregada com o objetivo principal de diminuir essas complicações, além de produzir maior conforto e redução dos custos de hospitalização, embora com maior curva de aprendizagem. O objetivo deste estudo foi avaliar a realização de coronariografia transradial, executada por operadores sem experiência na técnica, e comparar com a abordagem clássica (Sones e femoral) em termos de sucesso do procedimento e complicações, analisando a influência da curva de aprendizagem. MÉTODO: Estudo multicêntrico realizado em 14 hospitais do interior de São Paulo, no período de um ano. Foram randomizados mil pacientes de maneira equivalente para as técnicas transradial ou clássica. RESULTADOS: A taxa de sucesso em ambos os grupos foi similar (97,8% vs. 98,5%; P = 0,47). No grupo clássico, 95,2% dos procedimentos foram realizados pela técnica de Sones. O número de cateteres utilizados, a duração do procedimento e o tempo de exposição aos raios X foram maiores no grupo transradial (P < 0,001). Não houve diferença quanto às complicações maiores (morte, infarto e acidente vascular cerebral) e quanto às complicações vasculares e hemorrágicas. Os grupos que realizaram mais de 100 procedimentos pela artéria radial (3 centros/5 operadores) obtiveram menor taxa de insucesso (1,6% vs. 3,6%; P = 0,04). CONCLUSÃO: Um período de aprendizagem para a realização de procedimentos pela via transradial mostra-se necessário, porém não se acompanha de menor índice de sucesso, nem tampouco de maior taxa de complicações para os pacientes.

LESION OF THE ANTEROVENTRAL 3RD VENTRICLE REGION IMPAIRS THE RECOVERY OF ARTERIAL-PRESSURE INDUCED BY HYPERTONIC SALINE IN RATS SUBMITTED TO HEMORRHAGIC-SHOCK

The effect of intravenous infusion of hypertonic saline (HS, 7.5% NaCl) on the recovery of mean arteria pressure (MAP) after hemorrhage was studied in sham-operated rats and in rats with electrolytic lesion of the anteroventral third ventricle (AV3V) region (4 h, 4 and 20 days). Rats anesthetized with thiopental sodium were bled (about 2.8 ml/100 g) until the MAP was stabilized at the level of 60 mmHg for 30 min. In sham-lesioned rats, MAP increased to 90 mmHg and became stable near this level after intravenous infusion of 7.5% NaCl (4 ml/kg b.wt.). In AV3V-lesioned rats, the same infusion induced a smaller increase in MAP (80 mmHg) and the MAP returned to pre-infusion levels within 30 min. These results show that the AV3V region plays an important role in the recovery of arterial pressure induced by hypertonic saline in rats submitted to hemorrhagic shock.

Neo-clerodane diterpenoid, a new metalloprotease snake venom inhibitor from Baccharis trimera (Asteraceae): anti-proteolytic and anti-hemorrhagic properties

Many plants are used in traditional medicine as active agents against various effects induced by snakebite. Few attempts have been made however to identify the nature of plain natural products with anti-ophidian properties. Baccharis trimera (Less) DC (Asteraceae), known in Brazil as carqueja. has been popularly used to treat liver diseases. rheumatism. diabetes, as well as digestive, hepatic and renal disorders. The active component was identified as 7alpha-hydroxy-3,13-clerodadiene-16,15:18,19-diolide, C20H28O5, (clerodane diterpenoid, Bt-CD). We report now the anti-proteolytic and anti-hemorrhagic propenies against snake venoms of a Bt-CD inhibitor from B. trimera. Bt-CD exhibited full inhibition of hemorrhage and proteolytic activity caused by Bothrops snake venoms. The inhibitor was able to neutralize the hemorrhagic, fibrinogenolytic and caseinolytic activities of class P-I and III metalloproteases isolated from B. neuwiedi and B. jararacussu venoms. No inhibition of the coagulant activity was observed. Bt-CD also partially inhibited the edema induced by other crude venoms, metallopronteases, basic and acidic phospholipases A(2). To further elucidate the inhibitory specificity of Bt-CD against metalloproteases isolated from snake venoms, a deeper understanding of its Structure and function is necessary. Furthermore, the potential use of these inhibitors to complement anti-venom as an alternative treatment of snakebite envenomations needs to be evaluated in future Studies. (C) 2004 Elsevier B.V.. All rights reserved.