• 목록
• 답변
• 게시판 리스트 옵션
  • 수정
  • 삭제

E fraction F17 (18 mg) eluted with n-hexane/CH2Cl2 (9.7:0.3) and the fraction F76 (16 mg) eluted with n-hexane/CH2Cl2 (5:5) contained the pure halogenated monoterpenes 1 and 2, respectively. The fractions F112 to F118 eluted with n-hexane/CH2Cl2 (3.5:6.5) and F119-F125 eluted with n-hexane/CH2Cl2 (3:7) contained the pure cholesterol (42 mg). These fractions were also obtained according to the method by Fonseca et al. [31], and were analyzed according to Vasconcelos et al. [30] by HRGC/MS on a HP 5890 series GC system, coupled to a HP 5973 mass selective detector in the EI mode (70 eV) equipped withda Silva et al. Journal of Venomous Animals and Toxins including Tropical Diseases (2015) 21:Page 3 ofFigure 1 Chemical structure of (1) monoterpene 1 (8-bromo-3,4,7-trichloro-3,7-dimethyl-1E,5E-octadiene), (2) monoterpene 2 (1,8-dibromo-3,4,7-trichloro-3,7-dimethyl-1E,5E-octadiene) and of the (3) cholesterol fraction.a HP-1 MS capillary column (30 m ?0.25 mm, film thickness 0.25 m), and dissolved in DMSO (30 , v/v) as well.Venom and animalsfor 30 minutes at room temperature, and then, hemolytic activity was evaluated. Positive controls were performed by incubating venom with DMSO (5 v/v) or saline solution, instead of extracts. DMSO (5 v/v) or extracts alone were used 1-(4-Bromo-2-pyridyl)piperazine as negative controls.Anticoagulant activityB. jararaca Methyl 2-((4-nitro-1h-pyrazol-1-yl)methyl)benzoate venom was kindly supplied by the Ezequiel Dias Foundation (FUNED), Belo Horizonte, Minas Gerais state, Brazil, vacuum dried and stored at -20 until use. Male Balb/c mice (Mus musculus species) weighting 18?0 g were obtained from the Center of Laboratory Animals (NAL) of the Federal Fluminense University (UFF). They were housed under controlled conditions of temperature (24 ?1 ) and light. All the experiments performed were approved by the UFF Institutional Committee for Ethics in Animal PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/9638577 Experimentation (CEUA: 200/10) and were in accordance with the guidelines of the Brazilian Committee for Animal Experimentation (COBEA).Antihemolytic activityThe degree of hemolysis of B. jararaca venom PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/13485127 was determined by the indirect hemolytic test using human erythrocytes and hen's egg yolk emulsion as substrates [35]. The lowest amount of B. jararaca venom that produced 100 hemolysis was called the minimum indirect hemolytic dose (MIHD). Inhibitory experiments were performed by incubating algal extracts with one MIHDThe coagulant activity of B. jararaca venom was monitored using a digital Amelung coagulometer, model KC4A (Labcon, Germany). Different concentrations of B. jararaca venom were mixed with diluted citrated plasma (1:1 in saline) donated from healthy volunteers collected at a local public blood bank (University Hospital Ant io Pedro of UFF). The amount of venom (g/mL) that clotted plasma in 60 seconds was called the minimum coagulant dose (MCD). To evaluate the inhibitory effect, algal extracts were incubated for 30 minutes at room temperature with one gahmj.2015.132 MCD, and then, the mixture was added to plasma and coagulation time was recorded. Positive controls were performed in parallel by incubating venom with DMSO (1 v/v) or saline solution. DMSO (1 v/v) or extracts alone were used as negative controls.Antiproteolytic activityProteolytic activity of B. jararaca venom was determined using azocasein as substrate (0.2 w/v, in 20 mM Tris Cl, 8 mM CaCl2, pH 8.8) [36] ?with minor modifications. Anda Silva et al. Journal of Venomous Animals and Toxins including Tropical Diseases (2015) 21:Page 4 ofeffective concentration (EC) was defined.