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The Journal of Biological Chemistry Apr 1981The formation of a fibrin clot is initiated after the proteolytic cleavage of fibrinogen by thrombin. The enzyme removes fibrinopeptides A and B and generates fibrin...
The formation of a fibrin clot is initiated after the proteolytic cleavage of fibrinogen by thrombin. The enzyme removes fibrinopeptides A and B and generates fibrin monomer which spontaneously polymerizes. Polymerization appears to occur though the interaction of complementary binding sites on the NH2-terminal and COOH-terminal (Fragment D) regions of the molecule. A peptide has been isolated from the gamma chain remnant of fibrinogen Fragment D1 which has the ability to bind to the NH2-terminal region of fibrinogen as well as to inhibit fibrin monomer polymerization. The peptide reduces the maximum rate and extent of the polymerization of thrombin or batroxobin fibrin monomer and increases the lag time. The D1 peptide does not interact with disulfide knot, fibrinogen, or Fragment D1, but it binds to thrombin-treated disulfide knot with a Kd of 1.45 X 10(-6) M at approximately two binding sites per molecule of disulfide knot. Fibrin monomer formed either by thrombin or batroxobin binds approximately two molecules of D1 peptide per molecule of fibrin monomer, indicating that the complementary site is revealed by the loss of fibrinopeptide A. The NH2-terminal sequence (Thr-Arg-Trp) and COOH-terminal sequence (Ala-Gly-Asp-Val) of the D1 peptide were determined. Therefore the gamma 373-410 region of fibrinogen contains a polymerization site which is complementary to the thrombin-activated site on the NH2-terminal region of fibrinogen.
Topics: Amino Acids; Batroxobin; Fibrin; Fibrinogen; Fibrinolysin; Humans; Kinetics; Macromolecular Substances; Peptide Fragments; Plasminogen
PubMed: 6451630
DOI: No ID Found -
Batroxobin mobilizes circulating endothelial progenitor cells in patients with deep vein thrombosis.Clinical and Applied... Feb 2011Batroxobin, a thrombin-like enzyme from Bothrops atrox moojeni venom, is associated with the reduction of fibrinogen levels in plasma and the enhancement of...
Batroxobin, a thrombin-like enzyme from Bothrops atrox moojeni venom, is associated with the reduction of fibrinogen levels in plasma and the enhancement of anticoagulation and fibrinolysis. In this study, 15 patients with deep vein thrombosis (DVT) achieved successful limb salvage after the administration of batroxobin. We found that the levels of CD34+, CD31+, CD34+/CD31+, and vascular endothelial cadherin (VE-cadherin+) cells had increased in the peripheral blood of patients at 7 days and 14 days after treatment. At 0 day, 7 days, and 14 days, the percentages of CD34+ cells, which are assumed to be hematopoietic stem cells, are 0.39% ± 0.43%, 0.71% ± 0.50%, and 1.11% ± 0.66%, respectively. The levels of CD34+ cells at 14 days are significantly higher than the levels on the first day (P = .004). The levels of CD31+ cells and VE-cadherin+ cells, which represent mature endothelial cells, at 7 days (34.15% ± 11.32%, P = .013; 1.25% ± 1.39%, P = .014) and 14 days (35.21% ± 7.66%, P = .071; 1.85% ± 2.60%, P = .117) were slightly elevated compared with those at 0 day (27.55% ± 8.65%; 0.25 ± 0.39%). The double positive of CD34 and CD31 cells are assumed to be endothelial progenitor cells (EPCs). The levels of CD34+/CD31+ cells at 7 days (0.69% ± 0.50%, P = .001) and 14 days (1.07% ± 0.66%, P = .006) are significantly higher than that on the initial day (0.28% ± 0.30%). The number of CD34+/CD31+ cells significantly increased, indicating that in addition to its role in anticoagulation and fibrinolysis, treatment with batroxobin might simultaneously activate circulating EPCs that might promote the recanalization of the damaged vessel wall.
Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Batroxobin; Endothelial Cells; Female; Fibrinolytic Agents; Hematopoietic Stem Cell Mobilization; Humans; Male; Middle Aged; Radiography; Stem Cells; Time Factors; Venous Thrombosis
PubMed: 19825915
DOI: 10.1177/1076029609347903 -
Medicine Dec 2019Hemocoagulase is isolated and purified from snake venoms. Hemocoagulase agents have been widely used in the prevention and treatment of surgical bleeding. A systematic... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Hemocoagulase is isolated and purified from snake venoms. Hemocoagulase agents have been widely used in the prevention and treatment of surgical bleeding. A systematic review was performed to evaluate the effects of hemocoagulase on postoperative bleeding and transfusion in patients who underwent cardiac surgery.
METHODS
Electronic databases were searched to identify all clinical trials comparing hemocoagulase with placebo/blank on postoperative bleeding and transfusion in patients undergoing cardiac surgery. Two authors independently extracted perioperative data and outcome data. For continuous variables, treatment effects were calculated as weighted mean difference and 95% confidential interval (CI). For dichotomous data, treatment effects were calculated as odds ratio and 95% CI. Each outcome was tested for heterogeneity, and randomized-effects or fixed-effects model was used in the presence or absence of significant heterogeneity. Sensitivity analyses were done by examining the influence of statistical model and individual trial on estimated treatment effects. Publication bias was explored through visual inspection of funnel plots of the outcomes. Statistical significance was defined as P < .05.
RESULTS
Our search yielded 12 studies including 900 patients, and 510 patients were allocated into hemocoagulase group and 390 into control group. Meta-analysis suggested that, hemocoagulase-treated patients had less bleeding volume, reduced red blood cells and fresh frozen plasma transfusion, and higher hemoglobin level than those of controlled patients postoperatively. Meta-analysis also showed that, hemocoagulase did not influence intraoperative heparin or protamine dosages and postoperative platelet counts. Meta-analysis demonstrated that, hemocoagulase-treated patients had significantly shorter postoperative prothrombin time, activated partial thromboplastin time, and thrombin time, higher fibrinogen level and similar D-dimer level when compared to control patients.
CONCLUSION
This meta-analysis has found some evidence showing that hemocoagulase reduces postoperative bleeding, and blood transfusion requirement in patients undergoing cardiac surgery. However, these findings should be interpreted rigorously. Further well-conducted trials are required to assess the blood-saving effects and mechanisms of Hemocoagulase.
Topics: Batroxobin; Blood Transfusion; Cardiac Surgical Procedures; Hemostatics; Humans; Postoperative Hemorrhage
PubMed: 31876750
DOI: 10.1097/MD.0000000000018534 -
European Journal of Gastroenterology &... Dec 2021This study aimed to evaluate the efficacy and safety of using Hemocoagulase Bothrops Atrox in the submucosal injection solution for endoscopic submucosal dissection... (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
This study aimed to evaluate the efficacy and safety of using Hemocoagulase Bothrops Atrox in the submucosal injection solution for endoscopic submucosal dissection (ESD).
METHODS
A total of 120 patients with superficial neoplastic lesions of the esophagus, stomach, and colon receiving ESD were randomly divided into two groups: The epinephrine group used epinephrine-containing submucosal fluid cushion for ESD, while the hemocoagulase group used Hemocoagulase Bothrops Atrox-containing submucosal fluid cushion for ESD. The preoperative, intraoperative, and postoperative clinical parameters and postoperative adverse events of the two groups were recorded, and comparative analysis within and between groups was performed.
RESULTS
There was no significant difference in the demographic and clinical characteristics between the hemocoagulase and epinephrine group (all P > 0.05). ESD surgery was completed in all patients. The hemocoagulase group had significantly shorter surgery time (P = 0.003) and less number of intraoperative bleeding (P = 0.010) than the epinephrine group. However, there was no significant difference in the incidences of postoperative delayed hemorrhage, and adverse events between the two groups (all P > 0.05). Multivariate linear regression demonstrated that the epinephrine group had significantly more number of intraoperative bleeding (B: 0.98, 95% confidence interval: 0.04-1.93) as compared with the hemocoagulase group.
CONCLUSION
Compared with epinephrine, using Hemocoagulase Bothrops Atrox in the submucosal injection for ESD surgery can significantly reduce the number of intraoperative bleeding, shorten the operation time, and did not elevate the incidence of adverse events.
Topics: Animals; Batroxobin; Bothrops; Endoscopic Mucosal Resection; Epinephrine; Humans; Postoperative Hemorrhage; Stomach Neoplasms; Treatment Outcome
PubMed: 34034279
DOI: 10.1097/MEG.0000000000002206 -
World Journal of Gastroenterology Jul 2016To evaluate the hemostatic effect of topical hemocoagulase spray in digestive endoscopy. (Randomized Controlled Trial)
Randomized Controlled Trial
AIM
To evaluate the hemostatic effect of topical hemocoagulase spray in digestive endoscopy.
METHODS
Eighty-nine patients who developed oozing bleeding during endoscopic treatment from September 2014 to October 2014 at Center for Digestive Endoscopy, Tianjin Medical University General Hospital were randomly divided into either a study group (n = 39) or a control group (n = 50). The study group was given topical hemocoagulase spray intraoperatively, while the control group was given traditional 8% norepinephrine spray. Hemostatic efficacy was compared between the two groups. Bleeding site, wound cleanliness and perforation were recorded, and the rates of perforation and late bleeding were compared.
RESULTS
Successful hemostasis was achieved in 39 (100%) patients of the study group and in 47 (94.0%) patients of the control group, and there was no significant difference in the rate of successful hemostasis between the two groups. Compared with the control group, after topical hemocoagulase spray in the study group, the surgical field was clearer, the bleeding site was more easily identified, and the wound was cleaner. There was no significant difference in the rate of perforation between the study and control groups (16.7% vs 35.0%, P = 0.477), but the rates of late bleeding (0% vs 15.8%, P = 0.048) and overall complications (P = 0.032) were significantly lower in the study group.
CONCLUSION
Topical hemocoagulase spray has a definite hemostatic effect for oozing bleeding in digestive endoscopy, and this method is convenient, safe, and reliable. It is expected to become a new method for endoscopic hemostasis.
Topics: Administration, Topical; Batroxobin; Endoscopic Mucosal Resection; Endoscopy, Digestive System; Female; Hemostasis, Endoscopic; Hemostatics; Humans; Intestinal Perforation; Male; Middle Aged; Norepinephrine; Postoperative Complications; Postoperative Hemorrhage; Treatment Outcome; Vasoconstrictor Agents
PubMed: 27433096
DOI: 10.3748/wjg.v22.i25.5831 -
Heliyon Oct 2015To evaluate the in vivo efficacy of 915 MHz percutaneous coagulation in the treatment of hepatic artery injury.
OBJECTIVES
To evaluate the in vivo efficacy of 915 MHz percutaneous coagulation in the treatment of hepatic artery injury.
METHODS
After inducing hepatic artery injury, 8 dogs in each group underwent 915 MHz microwave percutaneous coagulation therapy and 8 dogs were injected with batroxobin and α-cyanoacrylate.
RESULTS
The hemostatic effects of 915 MHz microwave were better than drug injection, and the amount of bleeding was significantly lower (p < 0.05). Pathological examination showed that vessel wall necrosis were greater.
CONCLUSION
Contrast ultrasound guided 915 MHz microwave percutaneous coagulation treatment has potent hemostatic effects in the repair of in vivo hepatic artery injury.
PubMed: 27441219
DOI: 10.1016/j.heliyon.2015.e00030 -
Journal of Thrombosis and Haemostasis :... May 2018
An international collaborative study to calibrate the WHO 2nd International Standard for Ancrod (15/106) and the WHO Reference Reagent for Batroxobin (15/140): communication from the SSC of the ISTH.
Topics: Ancrod; Batroxobin; Calibration; Fibrinolysis; Fibrinolytic Agents; Humans; International Cooperation; Observer Variation; Predictive Value of Tests; Reference Standards; Reproducibility of Results; Thrombin Time; World Health Organization
PubMed: 29607604
DOI: 10.1111/jth.13996 -
The Journal of Biological Chemistry Jun 1988We have isolated and analyzed the gene for batroxobin, a thrombin-like snake venom enzyme. Three overlapping DNA segments containing the entire batroxobin gene were...
We have isolated and analyzed the gene for batroxobin, a thrombin-like snake venom enzyme. Three overlapping DNA segments containing the entire batroxobin gene were identified. Sequence analysis revealed that the batroxobin gene spans 8 kilobase pairs and contains five exons. Mature batroxobin is encoded by four separate exons, 2 to 5. The catalytic residues of batroxobin, His-41, Asp-86, and Ser-178, are encoded by separate exons, exons 2, 3, and 5, respectively. The exon/intron organization of the batroxobin gene is different from that of the prothrombin gene but very similar to those of the trypsin and kallikrein genes. These results indicate that batroxobin is not a member of the prothrombin family but one of the trypsin/kallikrein family. The snake venom gland is assumed to originate from the submaxillary gland. Therefore, batroxobin is expected to be a member of the glandular kallikrein family.
Topics: Amino Acid Sequence; Base Sequence; Batroxobin; Crotalid Venoms; DNA; Exons; Introns; Kallikreins; Molecular Sequence Data; Serine Endopeptidases; Trypsin
PubMed: 3163691
DOI: No ID Found -
Biophysical Journal May 1982Fibrinogen solutions (concentrations 2 mg/ml, 0.15-M Tris-NaCl buffer, pH 7.4) were incubated at 20 degrees C with quantities of reptilase or thrombin that were so small...
Fibrinogen solutions (concentrations 2 mg/ml, 0.15-M Tris-NaCl buffer, pH 7.4) were incubated at 20 degrees C with quantities of reptilase or thrombin that were so small that the polymerization process could be followed for several hours by means of static and dynamic light scattering. The scattered intensity and its correlation function were recorded at scattering angles between 30 degrees and 150 degrees. The measured data were compared with model calculations based on the Flory-Stockmayer distribution, which predicts a sol-gel phase transition. This distribution is characterized by a parameter, lambda, that indicates the extent of aggregation. lambda = 0 corresponds to the monomeric solution, and lambda = 1 indicates the sol-gel transition. Good agreement was found for monomeric units of 75-nm length aggregating (a) end-to-end in the early stage (0 less than or equal to lambda less than or equal to 0.3), and (b) in a staggered overlap pattern for the progressing polymerization (0.3 less than or equal to lambda less than 1). Before the gel point was reached, no systemic difference was observed between the data obtained after activation with thrombin which releases both fibrinopeptides A and B from fibrinogen, and reptilase, which exclusively releases the fibrinopeptides A. This confirms that the release of the fibrinopeptides A is the essential prerequisite for the aggregation process.
Topics: Batroxobin; Fibrin; Fibrinogen; Humans; Light; Macromolecular Substances; Mathematics; Polymers; Scattering, Radiation; Thrombin
PubMed: 7093417
DOI: 10.1016/S0006-3495(82)84539-6 -
Se Pu = Chinese Journal of... Sep 2022Snake venom thrombin drugs are hemostatic drugs prepared from venom, and the main active ingredients are snake venom thrombin-like enzymes (svTLEs). The svTLEs derived...
[Determination of the species origin and thrombin-like enzyme content of venom by ultra-high performance liquid chromatography-tandem mass spectrometry based on marker peptide].
Snake venom thrombin drugs are hemostatic drugs prepared from venom, and the main active ingredients are snake venom thrombin-like enzymes (svTLEs). The svTLEs derived from different snake species differ in their structures, hemostatic mechanisms, and pharmacological effects. Therefore, accurate identification of the source of snake venom species and determination of the svTLE content are essential to ensure the quality of these products. Based on proteomics technology, the marker peptides of svTLEs from were screened with species specificity for the first time in this study, and an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for species identification and determination of the svTLE content of was established. After reductive alkylation and trypsin enzymolysis of the purified svTLE from , enzymatic peptide fragments were obtained and determined by easy-nano liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (Nano LC-Q-Exactive-MS). The mass spectrum data were analyzed by Proteome Discoverer 2.2 software. The maker peptide "EAYNGLPAK", which characterized the svTLE from , was finally screened and validated by comparison of the basic local alignment search tool (BLAST) with the NCBI and UniProt databases. For the marker peptide, the enzymolysis temperature, enzymolysis time and amount of enzyme for the sample preparation were optimized. The optimized enzymolysis conditions were as follows: enzymolysis temperature, 37 ℃; enzymolysis time, 4 h; and amount of enzyme, 10 μL. A qualitative and quantitative detection method based on UHPLC-MS/MS was established by optimizing the chromatographic and mass spectrometric conditions. Accordingly, 20 mg of the evenly mixed sample was weighed and placed in a 100 mL volumetric flask. Then, 25 mmol/L ammonium bicarbonate solution was added to dissolve the sample, and the solution was diluted to the scale. Precisely 1.00 mL of the solution was extracted; subsequently, addition of 10 μL trypsin solution was added, followed by shaking, and the mixture was placed in an incubator for 4 h to induce enzymolysis at a constant temperature of 37 ℃. The mixture was subsequently removed from the incubator, cooled to ambient temperature, centrifuged at 12000 r/min for 10 min, and analyzed by LC-MS. Separation was performed on the UPLC system with a Thermo Hypersil GOLD C18 column (100 mm×2.1 mm, 3.0 μm) under the gradient elution of acetonitrile containing 0.1% (v/v) acetic acid and water containing 0.1%(v/v) acetic acid, at a flow rate of 0.3 mL/min, column temperature of 30 ℃, and injection volume of 2 μL. The maker peptides were determined in the electrospray positive ionization (ESI) and multiple reaction monitoring (MRM) modes using the external standard curve method. The detection ions were 481.9> 315.2 and 481.9> 485.2. There was a good linear relationship between the mass concentration of the marker peptide and the chromatographic peak area in the range of 2.5-30 ng/mL, and the correlation coefficient () was 0.9996, The limit of detection (=3) and limit of quantification (=10) were 2.5 mg/kg and 6.25 mg/kg, respectively. At spiked levels of 40, 80, and 120 mg/kg, the recoveries of the marker peptides were 95.5%-101.9%, while the relative standard deviations (RSDs) of the results for parallel analyses at various spiked levels were 1.1%-3.2%. The developed method is simple, rapid, sensitive, and specific, and it can be used for the identification of venom species and determination of the svTLE content. The findings of this study would help ensure the quality of hemocoagulase products from the relevant source and provide a reference for the quality control of other snake venom products.
Topics: Acetonitriles; Animals; Batroxobin; Bothrops; Chromatography, High Pressure Liquid; Hemostatics; Peptide Fragments; Peptides; Proteome; Snake Venoms; Tandem Mass Spectrometry; Thrombin; Trypsin; Water
PubMed: 36156627
DOI: 10.3724/SP.J.1123.2021.12020