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A myotoxin from Bothrops atrox snake venom was purified by cationic exchange on CM-Sephadex C-50 with 0,05M ammonium acetate pH 7. The myotoxin is a basic protein and by gel filtration and PAGE-SDS was demonstrated that protein has a molecular weight of 27 kDa and two polipeptides chain of 14 kDa each one. The inoculation of myotoxin in gastrocnemius muscle of white mice produce liberation of creatin kinase as well as myonecrosis. The myotoxin has phospholipasic, anticoagulant and edematic activity, but not hemolytic activity.

The myotoxin isolated from the venom of the snake Bothrops brazili has been studied in its mode of action. In the first hour of activity the myotoxin injected in the gastrocnemius muscle of white mice, induces the release of creatine kinase and lactate dehydrogenase, while by PAGE-SDS, it is shown that the incubation of miotoxin with isolated gastrocnemius muscle releases other proteins from this tissue. Moreover myotoxin produces hipercontraction, delta lesions and increases the concentration of intramuscular calcium “in vivo” and “in vitro”, which does not depend on extracellular calcium entry through dyhidropyridine receptors. This increase of calcium would explain the hipercontraction observed. Also it could trigger the activation of endogenous proteases and lipases, which would lead to muscle necrosis.

A myotoxin from the venom of the snake Bothrops brazili has been purified by ion-exchange chromatography on CM-Sephadex C-50 with 0,05 M ammonium acetate buffer pH 7. The homogeneity was evaluated by PAGE with and without SDS, immunodiffusion and immunoelectrophoresis. The myotoxin is a basic protein with 15,6% of Lys+Arg; it is not a glicoprotein, has not enzymatic activity, and corresponds to 25% of the whole venom protein. The molecular weight of the myotoxin was determined by PAGE-SDS and gel filtration chromatography. The myotoxin has 30 KDa of molecular weight and two polypeptide chains of 15 KDa each. Myotoxin produces a severe necrosis on the gastrocnemius muscle of white mice. The myotoxin does not have hemolytic nor anticoagulant activity. However, produces edema with a DEM of 32,6 mg of protein.

A proteolitic enzyme was purified from Bothrops brazili peruvian snake venom using Sephadex G-100 followed by CM-Sephadex C-50, in both two cases with 0.05M ammonium acetate pH 7,0. The enzyme was purified 3,2 fold with 52,5% of yield and by gel filtration the enzyme showed 18 000 of molecular weight, while the PAGE-SIDS showed only one protein band of 22 000 in the presence of mercaptoethanol and 20 300 under nonreducing conditions which suggests that the enzyme has a single polypeptide chain with disulfide bond. The enzyme hydrolizes fibrinogen, fibrin, casein and albumin, but not hemoglobin and mioglobin. The enzyme is a Aa-fibrinogenase because preferentially hydrolizes the Aa chain of the fibrinogen molecule. This activity is inhibited by EDTA but not by PMSF, TLCK, iodoacetate and pepstatin, suggests that is a metalloproteinase, however the ions Ca++, Mg++ and Zn++ cannot reactivat...

Snake venoms are polienzymatic concentrated whose biological activity on some bacteria and protozoa has been proven. To study the in vitro activity of Lachesis muta and B othrops atrox venoms on the viability and the development of Ascaris suum eggs is the main objecbve of this work. The venoms were employed in non embryonated and in vitro embryonated eggs of Ascaris suum at different concentrations (2, 4, 8,16 mg/mL). The activity of the venoms in the eggs and other substances like 5,25% sodium hypochlorite, Albendazol (commercial solution) and sodium chloride was compared. Both venoms, at 2, 4, 8, 16 mg/mL concentrations, inhibited segmentation of these eggs until the sixth day of incubation, when they began an apparently normal embryonation process that ended in the formation of the infective stage. The most effective inhibitory concentration was 16 mg/mL of B. atrox venom. The sodium...

Phospholipase A activity was evaluated into six snake venoms. Spectrophotometrical method adapted for our lab conditions previously and delay lipoprotein coagulation macroscopical method were compared. In both cases, the phospholipase activity was registrated in the venom of Micrurus spixii, Crotalus durissus, Bothrops brazili Lachesis muta and Bothrops atrox in the same decrease order. However the enzymatic activity orily was detected in the venom of Mícrurus surinamensis with the spectrophotometrical method. In addition the results related to amounth venom with each technique achieved the major sensitive of spectrophotometrical method than macroscopical technique because of 2 to 10 minor folds are required.

Objective: To characterize a hemorrhagic protein from Bothrops brazili snake venom and to determine if the polyvalent antibotropic serum is able to neutralize it. Material and Methods: A hemorrhagic protein from Bothrops brazili snake venom was purified through two chromatographical steps: Sephadex G-100 and CM Sephadex C-50, respectively, using 0,05M ammonium acetate buffer pH 7. In the last chromatographical system the protein was eluted after 0,3M sodium chloride was applied; thus, a unique band was achieved by PAGE-SDS. A hemorrhagic action monitored through caseinolytic activity obtained 8,4 folds of purification and the minimum hemorrhagic dose (MHD) was 6,61 ug in albine mice. Results: A structural analysis of associated carbohydrates showed 8,05% of hexoses, 11,62% of hexosamines, and 0,69% of sialic acid; its termostability was detected at 50°C for 10 minutes while total inhibi...

A myotoxin from Bothrops atrox snake venom was purified by cationic exchange on CM-Sephadex C-50 with 0,05M ammonium acetate pH 7. The myotoxin is a basic protein and by gel filtration and PAGE-SDS was demonstrated that protein has a molecular weight of 27 kDa and two polipeptides chain of 14 kDa each one. The inoculation of myotoxin in gastrocnemius muscle of white mice produce liberation of creatin kinase as well as myonecrosis. The myotoxin has phospholipasic, anticoagulant and edematic activity, but not hemolytic activity.

The myotoxin isolated from the venom of the snake Bothrops brazili has been studied in its mode of action. In the first hour of activity the myotoxin injected in the gastrocnemius muscle of white mice, induces the release of creatine kinase and lactate dehydrogenase, while by PAGE-SDS, it is shown that the incubation of miotoxin with isolated gastrocnemius muscle releases other proteins from this tissue. Moreover myotoxin produces hipercontraction, delta lesions and increases the concentration of intramuscular calcium “in vivo” and “in vitro”, which does not depend on extracellular calcium entry through dyhidropyridine receptors. This increase of calcium would explain the hipercontraction observed. Also it could trigger the activation of endogenous proteases and lipases, which would lead to muscle necrosis.

A myotoxin from the venom of the snake Bothrops brazili has been purified by ion-exchange chromatography on CM-Sephadex C-50 with 0,05 M ammonium acetate buffer pH 7. The homogeneity was evaluated by PAGE with and without SDS, immunodiffusion and immunoelectrophoresis. The myotoxin is a basic protein with 15,6% of Lys+Arg; it is not a glicoprotein, has not enzymatic activity, and corresponds to 25% of the whole venom protein. The molecular weight of the myotoxin was determined by PAGE-SDS and gel filtration chromatography. The myotoxin has 30 KDa of molecular weight and two polypeptide chains of 15 KDa each. Myotoxin produces a severe necrosis on the gastrocnemius muscle of white mice. The myotoxin does not have hemolytic nor anticoagulant activity. However, produces edema with a DEM of 32,6 mg of protein.

Se ha aislado una enzima proteolítica del veneno de la serpiente peruana Bothrops brazili, por cromatografía en Sephadex G-100 y CM-Sephadex C-50, con buffer acetato de amonio 0,05M pH 7,0. La enzima fue purificada 3,2 veces con un rendimiento de 52,5% y el peso molecular calculado por filtración en gel fue de 18 000, mientras que la PAGE-SDS permitió observar una sola banda proteica de 22 000 daltons en condiciones reductoras y de 20 300 daltons en condiciones no reductoras, determinándose que la enzima es de una sola cadena polipeptídica con al menos un enlace disulfuro. La enzima hidroliza fibrinógeno, fibrina, caseína y albúmina, pero no hemoglobina ni mioglobina. Por su acción sobre el fibrinógeno, es una Aa-fibrinogenasa ya que hidroliza primero la cadena Aa del fibrinógeno y luego la cadena Bb. Esta actividad es inhibida por EDTA pero no por PMSF, TLCK, iodoacetato y p...

Snake venoms are polienzymatic concentrated whose biological activity on some bacteria and protozoa has been proven. To study the in vitro activity of Lachesis muta and B othrops atrox venoms on the viability and the development of Ascaris suum eggs is the main objecbve of this work. The venoms were employed in non embryonated and in vitro embryonated eggs of Ascaris suum at different concentrations (2, 4, 8,16 mg/mL). The activity of the venoms in the eggs and other substances like 5,25% sodium hypochlorite, Albendazol (commercial solution) and sodium chloride was compared. Both venoms, at 2, 4, 8, 16 mg/mL concentrations, inhibited segmentation of these eggs until the sixth day of incubation, when they began an apparently normal embryonation process that ended in the formation of the infective stage. The most effective inhibitory concentration was 16 mg/mL of B. atrox venom. The sodium...

Phospholipase A activity was evaluated into six snake venoms. Spectrophotometrical method adapted for our lab conditions previously and delay lipoprotein coagulation macroscopical method were compared. In both cases, the phospholipase activity was registrated in the venom of Micrurus spixii, Crotalus durissus, Bothrops brazili Lachesis muta and Bothrops atrox in the same decrease order. However the enzymatic activity orily was detected in the venom of Mícrurus surinamensis with the spectrophotometrical method. In addition the results related to amounth venom with each technique achieved the major sensitive of spectrophotometrical method than macroscopical technique because of 2 to 10 minor folds are required.

An L-aminoacid oxidase was purified from Bothrops brazili snake venom using Sephadex G-100 and CM-Sepahdex C?50 chromatographic method; in both cases 0,1M ammonium acetate pH 6 was used as a eluted. The enzyme was purified 29,3 fold with 30,9% of yield. A homogeneus protein band was achieved by PAGE-SDS, inmunodifusion as well as immunoelectrophoresis. The enzyme was an acid glicoprotein with a molecular weight of 125,7 kd consisting of two subunits of 59,91 kd each, linked by noncovalent bonds. The optimum pH was in the range of 7,5 to 9,0 depending on the L-aminoacid used as substrate. It was a thermoestable protein untill 55 oC, but its activity decreased by alcaline treatment. On the other hand, antibacterial assays using the Grove's method demonstrated that the whole venom as well as its purified enzyme produced a severe action on standardized grown cultures of Staphylococcus aureus...

An L-aminoacid oxidase was purified from Bothrops brazili snake venom using Sephadex G-100 and CM-Sepahdex C?50 chromatographic method; in both cases 0,1M ammonium acetate pH 6 was used as a eluted. The enzyme was purified 29,3 fold with 30,9% of yield. A homogeneus protein band was achieved by PAGE-SDS, inmunodifusion as well as immunoelectrophoresis. The enzyme was an acid glicoprotein with a molecular weight of 125,7 kd consisting of two subunits of 59,91 kd each, linked by noncovalent bonds. The optimum pH was in the range of 7,5 to 9,0 depending on the L-aminoacid used as substrate. It was a thermoestable protein untill 55 oC, but its activity decreased by alcaline treatment. On the other hand, antibacterial assays using the Grove's method demonstrated that the whole venom as well as its purified enzyme produced a severe action on standardized grown cultures of Staphylococcus aureus...

A hemorrhagin with metalloprotease activity was purified from the venom of Bothrops pictussnake using Sephadex G-75 molecular gel filtration and DEAE A-50 ionic exchange column.Thus a homogeneus protein entity was obtained with 62 kDa under non reducting conditions.Hemorrhagin is a acid protein, attack both casein and collagen being 0,226 μg as a DHM.Chelating agent such as EDTA as well as 2 ß-mercaptoetanol and DTT produced stronginhibition both caseinolitic and hemorrhagic activities. Optimus pH was 7,5 and heatingtreatment reduced both activities, At 55 °C recovered activity on casein was 30,4%. On theother hand hemorrhagin is an antigenic entity showed on double immunodiffusion test andwas neutralizated fulling with 0,5, 1 and 2 doses of antivenom.