, 2000) The proteolytic activity of the venom was determined as

, 2000). The proteolytic activity of the venom was determined as 0.46 U/mg min and showed a dose dependent pattern ( Fig. 1A). This activity has already

been investigated for the H. lunatus venom, by Escobar and BMN 673 nmr co-workers in 2002. However, in this study, no proteolytic activity was found. This difference may be due to the method used in the previous work that may not have a comparable sensitivity to the method chosen in this study. Enzymes with gelatinolytic activity were detected in T. bahiensis and T. serrulatus venom ( Almeida et al., 2002). For these species Magalhães (1946) suggested that proteolytic enzymes were present based on observations of necrosis, hemolysis and gangrene following experimental injection in different animals. Necrosis is reported as one of the symptoms in humans stung by H. lunatus scorpions

( Anales de la Facultad de Medicina, 1967). The role for proteolytic enzymes in scorpion venoms is not very clear, but they may be related to tissue permeability, acting as spreading factors for the venom. These enzymes may also have a direct toxic effect in the development of pancreatitis presented by some envenomed patients ( Almeida et al., 2002; Renner et al., 1983). The H. lunatus venom also displays phospholipase and hyaluronidase activities (Figs. 1B and 2). The phospholipase CHIR99021 activity was examined through an indirect hemolytic assay, as described by Gutierrez in 1988 and is dose Phosphatidylinositol diacylglycerol-lyase dependent. The minimum phospholipasic dose (MPD), which is the dose capable of producing a 1 cm halo, was defined as 0.125 μg of soluble

venom. Phospholipase activity has been described for some scorpion venoms ( Schwartz et al., 2008), and phaiodactylipin was the first molecule from the venom of a scorpion belonging to the Iuridae family to be described ( Valdez-Cruz et al., 2007). This scorpion phospholipase belongs to the A2 family, class III. It is a heterodimeric protein with neurotoxic activity that acts by inhibiting the ryanodine receptor ( Zamudio et al., 1997). Besides their importance in digestion, venom phospholipases may also be related to other toxic symptoms such as inflammation, platelet aggregation, mionecrosis, hemolysis, neurotoxicity and cardiotoxicity ( Lambeau and Lazdunski, 1999). The hyaluronidase activity appears ubiquitous in all venom samples obtained from scorpion venoms ( Pessini et al., 2001; Seyedian et al., 2010). Using the zymogram technique ( Cevallos et al., 1992), we have identified a component with a molecular mass corresponding to 40 kDa as being responsible for the hyaluronidase activity of H. lunatus venom. The presence of hyaluronidase in scorpion venoms is probably related to the spread and absorption of venom’s toxic components and may affect the stability of blood vessel walls ( Veiga et al., 2001; Seyedian et al., 2010).

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