(Squamata: Viperidae) is the most important venomous snake in Central America,

(Squamata: Viperidae) is the most important venomous snake in Central America, being responsible for the majority of snakebite incidents. 2007 for Central America ranged from 193 to 1 1,461. Since snakebites impact, in most cases, poor people living in rural parts of tropical countries [3], the World Health Business (WHO), integrated snakebite envenoming in its list of neglected diseases (http://www.who.int/neglected_diseases/diseases/en/). Lance-headed pit vipers belonging the Viperidae family, especially the is responsible for 90% of all snakebites instances of major medical importance [4, 6]. is able to inoculate a relative large amount of venom and is considered extreme aggressive, being able to cause severe incidents [7]. 55778-02-4 manufacture Number 1 induces prominent local tissue damage, characterized by Rabbit Polyclonal to NF-kappaB p105/p50 (phospho-Ser893) swelling, blistering, prominent oedema, haemorrhage, dermonecrosis, and myonecrosis with medical manifestations that include bleeding, effects on platelet aggregation, coagulopathy, hypotension, hemodynamic alterations, pulmonary oedema, and acute renal failure. Additional less common effects include intravascular haemolysis, acute myocardial damage multiple organ failure, and death. The clinical features of the envenomation are affected by the venom parts, which vary relating to snake varieties, geographic region, age, sex, and environment [5, 8, 9]. Snake venoms are characterized like a complex mixture of bioactive molecules, which proteins compose more than 90% of the venom dry weight [10C12]. Many of these proteins are enzymes, in which the most abundant are phospholipases A2 (PLA2s; E.C.3.1.1.4) [10]. PLA2s are users of a protein superfamily that comprise several groups of enzymes with different catalytic mechanisms, as well as different practical and structural features, that cleavage the sn-2 acyl ester relationship of glycerolphospholipids generating free fatty acids and lysophospholipids [13, 14]. Snake venom PLA2s (svPLA2s) have been grouped into four classifications relating to small structural variations as group I and II, both sub-classified as type A or B. The group II is found in venoms from Viperidae family, while the group I is found in Elapidae and Hydrophiidae venoms [14]. svPLA2s from Viperidae family are placed into group IIB and are primarily subdivided in two types: Asp49 PLA2s, which have an Asp residue at position 49, and Lys49 PLA2s, showing a Lys residue at position 49. Different from Asp49 PLA2s, Lys49 PLA2s have low or any catalytic activity upon artificial substrates [13, 15, 16]. This present paper explains the biochemical and toxicological characterization of crude snake venom was collected from adult specimens, captured in Caldera and Gomez (Provence of Chiriqu, Panama) and in Arraijn (Provence of Panama, Panama). The snakes were maintained inside a serpentarium in the Gamboa Rainforest Vacation resort, Panam, where the crude venom was acquired by inducing the snake to bite a parafilm-wrapped jar. Venoms were centrifuged at 1,000?xg for 15?min, and supernatants were lyophilized and stored at ?20C in Microbiology Division at the Medicine Faculty of Panama University or college 55778-02-4 manufacture until used. Male albino Swiss mice, weighing 18C22?g, were utilized for the assays. The murine macrophage cell lines (J774A.1) were from Rio de Janeiro Cell Lender Collection 55778-02-4 manufacture (Brazil). RPMI-1640, penicillin, streptomycin, and L-glutamine were purchased from Sigma-Aldrich (MO, USA); fetal bovine serum (FBS) was from Cultilab (Brazil). All reagents were low endotoxin or endotoxin-free marks. Animal care was in accordance with the guidelines of the Brazilian College for Animal Experimentation (COBEA) and was authorized by the Committee for Ethics in Animals Utilization of Universidade de S?o Paulo (CEUA no. 06.1.291.53.3). CM-Sepharose and Phenyl-Sepharose resins were purchased from Amersham Biosciences, Uppsala, Sweden. The Kit CK-UV- Kinetic was purchased from Bioclin, Brazil. The following reagents: ethylenediaminetetraacetic acid (EDTA), molecular excess weight protein requirements, and acrylamide were from Sigma Chemical Co. All other chemicals reagents were of analytical grade from Merck, Aldrich or Pharmacia Biotech. 2.2. Purification and Biochemical Characterization of PLA2s by indirect erythrocyte lysis in agar comprising human being.

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