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Sample records for aprotinin

  1. Anaphylactic reaction after systemic application of aprotinin triggered by aprotinin-containing fibrin sealant.

    Science.gov (United States)

    Kober, Benjamin J; Scheule, Albertus M; Voth, Vladimir; Deschner, Norbert; Schmid, Eckhard; Ziemer, Gerhard

    2008-08-01

    We report a 67-yr-old male after multiple surgical procedures for treatment of arterial occlusive disease who suffered an anaphylactic reaction after administration of aprotinin (Trasylol) prior to urgent coronary artery bypass surgery. The patient had been treated with aprotinin-containing fibrin sealant in 2004 and in 2007, 2 wk before coronary artery bypass surgery. The postoperative serologic screening revealed positive results for qualitative aprotinin-specific IgG, highly elevated quantitative aprotinin-specific IgG and moderately elevated aprotinin-specific IgE antibodies.

  2. Aprotinin and hemostasis monitoring concerns during cardiac surgery.

    Science.gov (United States)

    Swartz, Michael F; Fink, Gregory W; Searles, Bruce

    2004-12-01

    Aprotinin (Trasylol) is a serine protease inhibitor, isolated from bovine lung that initially was marketed for the treatment of pancreatitis. In the mid 1980s, reports of its ability to decrease hemorrhaging after cardiopulmonary bypass surgery introduced the drug to the realm of cardiac surgery. Unfortunately, its introduction into this arena was followed by the publication of multiple studies and case reports that blamed aprotinin for poor outcomes in the form of early graft closure. More than 17 years have passed since the initial article describing the use of aprotinin during cardiopulmonary bypass, and with time there has been a significant increase in scientific knowledge and clinical experience. Interestingly, modern literature does not support the dogma that aprotinin is a procoagulant. Aprotinin increases the activated partial thromboplastin time (aPTT), as well as the kaolin- and celite-activated clotting time (ACT), regardless of heparin. Aprotinin, because of its ability to inhibit kallikrein, has been found to decrease thrombin antithrombin III complexes, fibrin-split products, fibrinopeptide 1+2, prothrombin fragments, and all markers of thrombin formation. Some authors have suggested that it may have a synergistic effect with heparin to ensure graft patency. Anticoagulation monitoring during the use of aprotinin also has been developed based on early studies. Aprotinin administration does influence the results of various ACT tests, and consequently different methods of testing anticoagulation have been developed. Researchers have demonstrated that the celite ACT is not "artificially" prolonged in the presence of heparin and aprotinin, rather the kaolin ACT is "artificially" shortened. This article will review the scientific literature with regard to aprotinin's anticoagulatory effects and review the current recommendations for hemostasis monitoring during the use of aprotinin.

  3. MONITORING OF ANTICOAGULATION IN APROTININ-TREATED PATIENTS DURING HEART OPERATION

    NARCIS (Netherlands)

    TABUCHI, N; NJO, TL; TIGCHELAAR, [No Value; HUYZEN, RJ; BOONSTRA, PW; VANOEVEREN, W

    1994-01-01

    Since aprotinin has become extensively used during cardiopulmonary bypass the maintenance of safe anticoagulation is a concern. Aprotinin affects anticoagulation measurement by the activated clotting time. Therefore, a reliable new measurement is needed to monitor anticoagulation during cardiopulmon

  4. Aprotinin induced lipohypertrophy and glomerulonephritis in an insulin dependent diabetic.

    Science.gov (United States)

    Dandona, P; Mier, A; Boag, F; Chappell, M; Beckett, A G

    1985-07-01

    In an insulin dependent diabetic who was hyperglycaemic and ketotic despite 3,000 u of insulin injected subcutaneously in 2 divided doses daily, 50 u of intravenous insulin infused over 24 hr restored normal glucose homeostasis. A combination of insulin (800 u) and aprotinin (10,000 u) given twice daily also produced adequate glucose homeostasis for a period of 12 months. The patient then developed local hypertrophy of subcutaneous tissue at the injection site and her diabetic control deteriorated. Non-selective proteinuria followed and she developed nephrotic syndrome. Renal biopsy revealed a membraneous glomerulonephritis with subepithelial immune complexes, appearances consistent with a drug-induced glomerulonephritis. Withdrawal of aprotinin led to a gradual remission of nephrotic syndrome and proteinuria over several months. During this period, her diabetes was well controlled with continuous subcutaneous infusion of insulin at a dose of 500 u/24 hr. This case report demonstrates: the effective use of aprotinin for prolonged periods in insulin dependent diabetics with abnormal absorption of subcutaneously injected insulin; aprotinin induced lipohypertrophy which was not observed when insulin was injected alone; aprotinin-associated glomerulonephritis and nephrotic syndrome; the effective use of CSII--at higher insulin doses--in such patients with subcutaneous malabsorption of insulin.

  5. Aprotinin in orthotopic liver transplantation : Evidence for a prohemostatic, but not a prothrombotic, effect

    NARCIS (Netherlands)

    Molenaar, IQ; Legnani, C; Groenland, THN; Palareti, G; Begliomini, B; Terpstra, OT; Porte, RJ

    2001-01-01

    Aprotinin reduces blood transfusion requirements in orthotopic liver transplantation (OLT). Concern has been voiced about the potential risk for thrombotic complications when aprotinin is used. The aim of this study is to evaluate the effects of aprotinin on the two components of the hemostatic syst

  6. Increased incidence of acute kidney injury with aprotinin use during cardiac surgery detected with urinary NGAL

    DEFF Research Database (Denmark)

    Wagener, G.; Gubitosa, G.; Wang, S.;

    2008-01-01

    BACKGROUND: Use of aprotinin has been associated with acute kidney injury after cardiac surgery. Neutrophil gelatinase-associated lipocalin (NGAL) is a novel, very sensitive marker for renal injury. Urinary NGAL may be able to detect renal injury caused by aprotinin. This study determined...... if the use of aprotinin is associated with an increased incidence of acute kidney injury and increased levels of urinary NGAL. METHODS: In this prospective, observational study 369 patients undergoing cardiac surgery were enrolled. 205 patients received aprotinin and 164 received epsilon amino-caproic acid...... intraoperatively. Urinary NGAL was measured before and immediately after cardiac surgery and 3, 18 and 24 h later. The association of aprotinin use with the incidence of acute kidney injury (increase of serum creatinine >0.5 mg/dl) and NGAL levels was determined using logistic and linear regression models. RESULTS...

  7. Impact of aprotinin and renal function on mortality: a retrospective single center analysis

    Directory of Open Access Journals (Sweden)

    Von Visger Jon

    2011-08-01

    Full Text Available Abstract Background An estimated up to 7% of high-risk cardiac surgery patients return to the operating room for bleeding. Aprotinin was used extensively as an antifibrinolytic agent in cardiac surgery patients for over 15 years and it showed efficacy in reducing bleeding. Aprotinin was removed from the market by the U.S. Food and Drug Administration after a large prospective, randomized clinical trial documented an increased mortality risk associated with the drug. Further debate arose when a meta-analysis of 211 randomized controlled trials showed no risk of renal failure or death associated with aprotinin. However, only patients with normal kidney function have been studied. Methods In this study, we look at a single center clinical trial using patients with varying degrees of baseline kidney function to answer the question: Does aprotinin increase odds of death given varying levels of preoperative kidney dysfunction? Results Based on our model, aprotinin use was associated with a 3.8-fold increase in odds of death one year later compared to no aprotinin use with p-value = 0.0018, regardless of level of preoperative kidney dysfunction after adjusting for other perioperative variables. Conclusions Lessons learned from our experience using aprotinin in the perioperative setting as an antifibrinolytic during open cardiac surgery should guide us in testing future antifibrinolytic drugs for not only efficacy of preventing bleeding, but for overall safety to the whole organism using long-term clinical outcome studies, including those with varying degree of baseline kidney function.

  8. Aprotinin and transfusion requirements in orthotopic liver transplantation : a multicentre randomised double-blind study

    NARCIS (Netherlands)

    Porte, RJ; Molenaar, IQ; Begliomini, B; Groenland, THN; Januszkiewicz, A; Lindgren, L; Palareti, G; Hermans, J; Terpstra, OT

    2000-01-01

    Background Intraoperative hyperfibrinolysis contributes to bleeding during adult orthotopic liver transplantation. We aimed to find out whether aprotinin, a potent antifibrinolytic agent, reduces blood loss and transfusion requirements. Methods We did a randomised, double-blind placebo-controlled tr

  9. Chitosan-aprotinin coated liposomes for oral peptide delivery: Development, characterisation and in vivo evaluation.

    Science.gov (United States)

    Werle, Martin; Takeuchi, Hirofumi

    2009-03-31

    In order to improve the systemic uptake of therapeutic peptides/proteins after oral administration, the polymer-protease inhibitor conjugate chitosan-aprotinin was synthesised and polyelectrolyte complexes between negatively charged multilamellar vesicles (MLV) and positively charged chitosan-aprotinin conjugate were prepared. It could be demonstrated that chitosan-aprotinin was capable of significantly inhibiting Trypsin in vitro in concentrations of 0.05% and 0.1%, whereas no inhibition was observed in the presence of 0.1% chitosan. The size range of the prepared MLV was between 3 and 4.5microm and the initially negative zeta potential (ca. -90mV) of the core liposomes switched to a positive value after polymer coating (ca. +40mV). Confocal laser microscopy studies showed comparable mucoadhesive properties of chitosan-aprotinin coated MLV and chitosan coated MLV. In comparison to calcitonin in solution, the area above the blood calcium concentration-time curve (AAC) after oral administration of calcitonin loaded chitosan coated MLV to rats increased around 11-fold, and around 15-fold in the case of calcitonin loaded chitosan-aprotinin coated MLV. Data gained in the current study are believed to contribute to the development of novel polymer-protease inhibitor based delivery systems.

  10. (99m)Tc-aprotinin - optimisation and validation of radiolabelling kits for routine preparation for diagnostic imaging of amyloidosis

    DEFF Research Database (Denmark)

    Denholt, Charlotte; Gillings, Nic

    2016-01-01

    Technetium-99m aprotinin was prepared from an optimised radiolabelling kit formulation containing aprotinin, alkaline buffer and stannous chloride (reducing agent) and radiolabelled using (99m) Tc-pertechnetate. The labelling was achieved within 25 min, with radiochemical purities of >98%....

  11. 99mTc‐aprotinin – optimisation and validation of radiolabelling kits for routine preparation for diagnostic imaging of amyloidosis

    OpenAIRE

    2016-01-01

    Abstract Technetium‐99m aprotinin was prepared from an optimised radiolabelling kit formulation containing aprotinin, alkaline buffer and stannous chloride (reducing agent) and radiolabelled using 99mTc‐pertechnetate. The labelling was achieved within 25 min, with radiochemical purities of >98%.

  12. 99mTc‐aprotinin – optimisation and validation of radiolabelling kits for routine preparation for diagnostic imaging of amyloidosis

    Science.gov (United States)

    Gillings, Nic

    2016-01-01

    Abstract Technetium‐99m aprotinin was prepared from an optimised radiolabelling kit formulation containing aprotinin, alkaline buffer and stannous chloride (reducing agent) and radiolabelled using 99mTc‐pertechnetate. The labelling was achieved within 25 min, with radiochemical purities of >98%. PMID:26923297

  13. MECHANISM OF PRESERVING EFFECT OF APROTININ ON PLATELET FUNCTION DURING CARDIOPULMONARY BYPASS

    Institute of Scientific and Technical Information of China (English)

    黄惠民; 丁文祥; 苏肇伉; 张伟忠

    1992-01-01

    The deficiency of platelet function is the main defect of hemostatic mechanism during cardiopulmonary bypass (CPB), which attributed to the postoperative bleeding complication to a great extent. The proteinase inhibitor aprotinin was reported to have preserving effect on platelet adhesion during CPB. In this clinical reserch we found that CPB caused plasma alpha 2-antiplasmin decreasing, indicating the fibrinolytic system activation. Meanwhile, the ristocetin-induced aggregation declined to 39.6% and platelet GPIb decreased to 50% of preoperative value. However, by treatment with aprotinin, the plasma alpha 2-antiplasmin during CPB did not change, platelet aggregation was improved and platelet GPIb was preserved, and consequently resulted in a 46% lower blood loss postoperatively. These results confirmed that aprotinin could inhibit the fibrinolysis during CPB, and thus relieve the platelet damage and improve the postoperative hemostatic mechanism.

  14. A clinical study of aprotinin blood anesthesia used in the surgery of liver cancer

    Institute of Scientific and Technical Information of China (English)

    易斌; 陶国才; 毕敏; 刘怀琼

    2004-01-01

    Objective: To investigate the role of aprotinin blood anesthesia used in hepatotomy. Methods: Patients with liver cancer undergoing hepatotomy were divided into two groups. In experimental group (40 patients) a loading dose with 1112 EPU aprotinin and maintained by 278 EPU/h was used until 2 h after operation. The control group (42 patients)was treated with 0.9% normal saline. The venous blood was withdrew for blood routine, thrombelastography and coagulable test at the time of preinduced, 1 h, 2 h and 4 h following the operation beginning, 6 h and 12 h after operation. The change of TEG and coagulable profile were monitored during the whole surgery. The volume of blood transfusion and hemorrhage between two groups were compared. Results: After the usage of aprotinin, the preoperative hypercoagulability of the experimental group was remitted and the coagulative state was kept relatively stable during the operation. However, hypercoagulability of the control group aggravated following the operation beginning and some of them switched to hypocoagulability. The volumes and rates of hemorrhage and transfusion were smaller in the experimental group than in the control group. Conclusion: Aprotinin can stabilize the coagulable state, reduce the volumes and rates of hemorrhage and transfusion, and is worth using in the surgery of operations of liver cancer.

  15. APROTININ PRESERVES HEMOSTASIS IN ASPIRIN-TREATED PATIENTS UNDERGOING CARDIOPULMONARY BYPASS

    NARCIS (Netherlands)

    TABUCHI, N; HUET, RCG; STURK, A; EIJSMAN, L; WILDEVUUR, CRH

    1994-01-01

    Various clinical trials have shown that hemostasis is improved by the administration of aprotinin during cardiopulmonary bypass. However, this effect has not been proved for those patients treated preoperatively with aspirin. Therefore, a double-blind, placebo-controlled study was conducted to test

  16. The effect of aprotinin on hypoxia-reoxygenation-induced changes in neutrophil and endothelial function.

    LENUS (Irish Health Repository)

    Harmon, D

    2012-02-03

    BACKGROUND AND OBJECTIVE: An acute inflammatory response associated with cerebral ischaemia-reperfusion contributes to the development of brain injury. Aprotinin has potential, though unexplained, neuroprotective effects in patients undergoing cardiac surgery. METHODS: Human neutrophil CD11 b\\/CD18, endothelial cell intercellular adhesion molecule-1 (ICAM-1) expression and endothelial interleukin (IL)-1beta supernatant concentrations in response to in vitro hypoxia-reoxygenation was studied in the presence or absence of aprotinin (1600 KIU mL(-1)). Adhesion molecule expression was quantified using flow cytometry and IL-1beta concentrations by enzyme-linked immunosorbent assay. Data were analysed using ANOVA and post hoc Student-Newman-Keuls test as appropriate. RESULTS: Exposure to 60-min hypoxia increased neutrophil CD11b expression compared to normoxia (170+\\/-46% vs. 91+\\/-27%, P = 0.001) (percent intensity of fluorescence compared to time 0) (n = 8). Hypoxia (60 min) produced greater upregulation of CD11b expression in controls compared to aprotinin-treated neutrophils [(170+\\/-46% vs. 129+\\/-40%) (P = 0.04)] (n = 8). Hypoxia-reoxygenation increased endothelial cell ICAM-1 expression (155+\\/-3.7 vs. 43+\\/-21 mean channel fluorescence, P = 0.0003) and IL-1beta supernatant concentrations compared to normoxia (3.4+\\/-0.4 vs. 2.6+\\/-0.2, P = 0.02) (n = 3). Hypoxia-reoxygenation produced greater upregulation of ICAM- 1 expression [(155+\\/-3.3 vs. 116+\\/-0.7) (P = 0.001)] and IL-1beta supernatant concentrations [(3.4+\\/-0.3 vs. 2.6+\\/-0.1) (P = 0.01)] in controls compared to aprotinin-treated endothelial cell preparation (n = 3). CONCLUSIONS: Hypoxia-reoxygenation-induced upregulation of neutrophil CD11b, endothelial cell ICAM-1 expression and IL-1beta concentrations is decreased by aprotinin at clinically relevant concentrations.

  17. The evaluation of aprotinin contained preservation solution with a new animal model

    Institute of Scientific and Technical Information of China (English)

    FU Qing-lin; ZHANG Xin-zhong; HAN Pei-li; SHI Rui-feng

    2008-01-01

    Objective To explore the protective effect of aprotinin contained LPD ( low potassium dextran) solution via an in situ rabbit lung preservation model. Methods Thirty New Zealand rabbits were divided randomly into 3 groups, 10 in each group. In group A (control group), the left lung hilus was clamped without solution perfusion; In group B ( LPD group) and group C ( aprotinin group), the lungs were perfused with LPD solution and aprotinin contained LPD, respectively. The lungs in all groups were stored at 10 centigrade in a specially made lung preservation container for 2 hours and then unclamped the lung hilus to rcperfuse the lung for 2 hours. Pulmonary venous blood samples were collected at pre-clamping of lung hilus,5 minutes and 120 minutes after reperfusion for analysis of blood gas. Biopsy of lung tissue was excised for morphological examination at pre-unclamping of lung hilas and 2 hours after reperfusion. Examination of bronchoalveolar lavage fluid was taken for the evaluation of inflammation status. Results Pulmonary venous partial pressure of oxygen ( PvPO2) in the 3 groups at 5 minutes and 120 minutes after reperfusion were significantly higher than those before clamping of lung hilus,respectively. PvPO2 in group A and group B at 120 minutes after reperfusion were significantly higher than those at 5 minutes after reperfusion. There was no significant difference of PvPO2 in group C between 5 minutes and 120 minutes after reperfusion. PvPO2 in group C at 5 minutes and 120 minutes after reperfusion were significantly higher than those in group A and group B. The morphological lesion was more severe in group A and B than that in group C. The PMN percentage in bronchoalveelar lavage fluid in group A and B was significantly higher than that in group C. Conclusions The protective effect of aprotinin is obvious for lung protection in animal model. Aprotinin contained lung preservation solution is superior to LPD for lung protection.

  18. The effects of aprotinin on blood product transfusion associated with thoracic aortic surgery requiring deep hypothermic circulatory arrest.

    LENUS (Irish Health Repository)

    Seigne, P W

    2012-02-03

    OBJECTIVE: To compare the effects of aprotinin on blood product use and postoperative complications in patients undergoing thoracic aortic surgery requiring deep hypothermic circulatory arrest. DESIGN: A retrospective study. SETTING: A university hospital. PARTICIPANTS: Nineteen patients who underwent elective or urgent thoracic aortic surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The total number of units of packed red blood cells, fresh frozen plasma, and platelets was significantly less in the group that received aprotinin (p = 0.01, 0.04, and 0.01). The intraoperative transfusion of packed red blood cells and platelets, collection and retransfusion of cell saver, and postoperative transfusion of fresh frozen plasma were also significantly less in the aprotinin group (p = 0.01, 0.02, 0.01, and 0.05). No patient in either group sustained renal dysfunction or a myocardial infarction. Two patients who had not received aprotinin suffered from chronic postoperative seizures, and one patient who had received aprotinin sustained a perioperative stroke. CONCLUSIONS: Low-dose aprotinin administration significantly decreases blood product transfusion requirements in the setting of thoracic aortic surgery requiring deep hypothermic circulatory arrest, and it does not appear to be associated with renal or myocardial dysfunction.

  19. Aprotinin decreases the incidence of cognitive deficit following CABG and cardiopulmonary bypass: a pilot randomized controlled study.

    LENUS (Irish Health Repository)

    Harmon, Dominic C

    2012-02-03

    PURPOSE: Cognitive deficit after coronary artery bypass surgery (CABG) has a high prevalence and is persistent. Meta-analysis of clinical trials demonstrates a decreased incidence of stroke after CABG when aprotinin is administrated perioperatively. We hypothesized that aprotinin administration would decrease the incidence of cognitive deficit after CABG. METHODS: Thirty-six ASA III-IV patients undergoing elective CABG were included in a prospective, randomized, single-blinded pilot study. Eighteen patients received aprotinin 2 x 10(6) KIU (loading dose), 2 x 10(6) KIU (added to circuit prime) and a continuous infusion of 5 x 10(5) KIU.hr(-1). A battery of cognitive tests was administered to patients and spouses (n = 18) the day before surgery, four days and six weeks postoperatively. RESULTS: Four days postoperatively new cognitive deficit (defined by a change in one or more cognitive domains using the Reliable Change Index method) was present in ten (58%) patients in the aprotinin group compared to 17 (94%) in the placebo group [95% confidence interval (CI) 0.10-0.62, P = 0.005); (P = 0.01)]. Six weeks postoperatively, four (23%) patients in the aprotinin group had cognitive deficit compared to ten (55%) in the placebo group (95% CI 0.80-0.16, P = 0.005); (P = 0.05). CONCLUSION: In this prospective pilot study, the incidence of cognitive deficit after CABG and cardiopulmonary bypass is decreased by the administration of high-dose aprotinin.

  20. THE IMPAIRMENT OF PLATELET FUNCTION IN FIBRINOLYSIS AND PRESERVING EFFECT OF APROTININ

    Institute of Scientific and Technical Information of China (English)

    黄惠民; 丁文祥; 苏肇伉; 张伟忠

    1992-01-01

    Platelet adhesion depends on the platelet membrane glycoprotein Ib (GPIb) and plasma von Willebrand Factor (vWF), which can be reflected by ristocetin-induced aggregation. Here we report damage effect of fibrinolysis and preserving effect of aprotinin on platelet function. Addition of 40 U/ml urokinase and 0.3 U/ml plasmin to PRP or washed platelets made the ristocetin-induced aggregation decline to 31.6% and 38.5% of control value respectively. The extent of declining was positively correlated with the concentration of urokinase and plasmin. Meanwhile, the platelet GPIb decreased to 76.4% of control value. The results showed that the fibrinolysis impaired the platelet function and this effect may be associated with the hydrolysis of GPIb. Further research found that by adding the same dose of urokinase or plasmin to aprotinin-pretreated PRP or washed platelets, the aggregation did not change statistically and decrement of GPIb is much less marked. We concluded that the aprotinin could relieve the platelet dsfunction effectively by its inhibitory effect on fibrinolytic activity.

  1. The effect of aprotinin, tranexamic acid, and aminocaproic acid on blood loss and use of blood products in major pediatric surgery : A meta-analysis

    NARCIS (Netherlands)

    Schouten, Esther S.; van de Pol, Alma C.; Schouten, Anton N. J.; Turner, Nigel M.; Jansen, Nicolaas J. G.; Bollen, Casper W.

    2009-01-01

    Objective: Aprotinin reduces the blood loss and transfusion of blood products in children undergoing major surgery. Aprotinin has been associated with severe side effects in adults, and tranexamic acid and aminocaproic acid have been found to be safer alternatives in adults. This systematic review a

  2. Comparison of three dose regimens of aprotinin in infants undergoing the arterial switch operation

    Directory of Open Access Journals (Sweden)

    Verma Yashwant

    2010-01-01

    Full Text Available To determine the most effective dose regimen of aprotinin for infants undergoing arterial switch operation for transposition of the great arteries in reducing blood loss and postoperative packed red blood cell (PRBC requirements. A total of 24 infants scheduled for arterial switch operation for transposition of the great arteries were included in the study. The infants were randomly assigned to one of the three groups. Group I (n = 8 patients received aprotinin in a dose of 20,000 kallikrein inhibiting units (KIU/kg after induction of anesthesia, 20,000 KIU/kg was added to the pump prime, and 20,000 KIU/kg/hour infusion for three hours after weaning from bypass; group II (n = 8 patients received aprotinin 30,000 KIU/kg after induction of anesthesia, 30,000 KIU/kg was added to the pump prime and 30,000 KIU/Kg/hour infusion for three hours after weaning from bypass; group III patients (n = 8 received aprotinin 40,000 KIU/kg after induction of anesthesia, 40,000 KIU/kg was added to the pump prime and 40,000 KIU/kg/hour infusion for three hours after weaning from bypass. Postoperatively, the cumulative hourly blood loss and PRBC requirements were noted up to 24 hours from the time of admission in the intensive care unit (ICU. Use of blood and blood products were noted. Coagulation parameters such as hematocrit, activated clotting time (ACT, fibrinogen, prothrombin time (PT, international normalized ratio (INR, platelet count, and fibrin degradation products (FDP were investigated before cardiopulmonary bypass (CPB, after protamine administration, and at four hours postoperatively in the ICU. The number of infants reexplored for increased mediastinal drainage was recorded. Renal functions were monitored by measuring urine output (hourly and serum urea (mg% and serum creatinine (mg% at 24 hours. The sternal closure time was comparable in all the three groups. Cumulative blood loss (ml/kg/24 hours was greatest in group I (17.30 ± 7.7, least in group

  3. Requirements for transfusion and postoperative outcomes in orthotopic liver transplantation:A meta-analysis on aprotinin

    Institute of Scientific and Technical Information of China (English)

    Cun-Ming Liu; Jing Chen; Xue-Hao Wang

    2008-01-01

    AIM:To study the effect of aprotinin used in orthotopic liver transplantation (OLT) on the intraoperative requirement for blood products and on the incidence of laparotomy for bleeding,thrombotic events and mortality.METHODS:A systematic review of the literature in the electronic database Medline and the Clinic Trials Registry Database was performed.Literature that did not fit our study were excluded.Patients in the reviewed studies were divided into two groups; one group used aprotinin (aprotinin group) while the other did not (control group).The data in the literature that fit our requirements were recorded.Weighted mean differences (WMD) in the requirements for blood products between the aprotinin group and the control group were tested using a fixed effect model.A Z test was performed to examine their reliability; the Fleiss method of fixed effect model was used to analyze data on postoperative events,and odds ratios (ORs) were tested and merged.RESULTS:Seven citations were examined in our study.Among them,a requirement for blood products was reported in 4 studies including 321 patients,while postoperative events were reported in 5 studies including 477 patients.The requirement for red blood cells and fresh frozen plasma in the aprotinin group was statistically lower than that in the control group (WMD = -1.80 units,95% CI,-3.38 to -0.22; WMD = -3.99 units,95% CI,-6.47 to -1.50,respectively).However,no significant difference was indicated in the incidence of laparotomy for bleeding,thrombotic events and mortality between the two groups.Analysis on blood loss,anaphylactic reactions and renal function was not performed in this study due to a lack of sufficient information.CONCLUSION:Aprotinin can reduce the intraoperative requirement for blood products in OLT,and has no significant effect on the incidence of laparotomy for bleeding,thrombotic events and mortality.

  4. Protective effects of intraperitoneal vitamin C, aprotinin and melatonin administration on retinal edema during experimental uveitis in the guinea pig.

    Science.gov (United States)

    Kükner, A Sahap; Kükner, Aysel; Naziroğlu, Mustafa; Colakoğlu, Neriman; Celebi, Serdal; Yilmaz, Turgut; Aydemir, Orhan

    2004-01-01

    A considerable amount of clinical and experimental evidence exists suggesting the involvement of reactive oxygen substances (ROS) in the aetiology of uveitis. The activated phagocytic system of polymorphonuclear leucocytes in uveitis is involved in the generation of ROS. In addition to their direct free radical scavenging action, aprotinin, melatonin and vitamin C are known to protect against oedema formation and can preserve plasma membrane fluidity and free radical production. Histological changes in the retina that occur during uveitis are not well explained. The purpose of this study was to determine whether vitamin C, aprotinin and melatonin can protect the retina from damage accompanying experimental uveitis (EU). Thirty adult male guinea pigs were divided into five groups of six animals each. The first group was used as control. The right eyes of groups 2, 3, 4 and 5 received an intravitreal injection of bovine serum albumin for induction of experimental uveitis. At the same time and also on the consecutive third day, groups 3, 4 and 5 received intraperitoneal injections of vitamin C (ascorbic acid, 100 mg kg(-1) body wt), aprotinin (20,000 kIU kg(-1) body wt) and melatonin (10 mg kg(-1) body wt), respectively. The animals were killed on the sixth day. The average thickness of the retina and inner plexiform layer for each eye was measured in sagittal section near the optic nerve and expressed in microns. The thickness of the retina and inner plexiform layer in the control group was significantly (p vitamin C, group EU plus aprotinin, group EU plus melatonin (p vitamin C, group EU plus aprotinin and group EU plus melatonin were significantly (p 0.05). In conclusion, this study demonstrated that oedematous effects of EU on the retina were reduced by the administration of intraperitoneal vitamin C, aprotinin and melatonin, i.e. these antioxidants had significant protective effects on the retina of guinea pigs against oedematous damage in EU. However, the

  5. PREOPERATIVE THERAPY OF LOW-DOSE ASPIRIN IN INTERNAL MAMMARY ARTERY BYPASS OPERATIONS WITH AND WITHOUT LOW-DOSE APROTININ

    NARCIS (Netherlands)

    SCHONBERGER, JPAM; BREDEE, JJ; VANOEVEREN, W; VANZUNDERT, AAJ; VERKROOST, M; TERWOORST, J; BAVINCK, JH; BERREKLOUW, E; WILDEVUUR, CRH

    1993-01-01

    The effect of preoperative low-dose aspirin (1 mg/kg of body weight) and intraoperative low-dose aprotinin (2 million kallikrein inactivator units) treatment on perioperative blood loss and blood requirements in patients who undergo internal mammary artery bypass operations is unknown. Therefore, we

  6. The urinary excretion of epidermal growth factor in the rat is reduced by aprotinin, a proteinase inhibitor

    DEFF Research Database (Denmark)

    Jørgensen, P E; Raaberg, Lasse; Poulsen, Steen Seier

    1990-01-01

    The present study on the rat shows that i.v. administration of the proteinase inhibitor aprotinin reduces the urinary output of immunoreactive epidermal growth factor (EGF) while the amount of immunoreactive EGF in the kidneys is increased. This indicates that the EGF-precursor in the rat kidney ...

  7. Purification of recombinant aprotinin produced in transgenic corn seed: separation from CTI utilizing ion-exchange chromatography

    Directory of Open Access Journals (Sweden)

    A. R. Azzoni

    2005-09-01

    Full Text Available Protein expression in transgenic plants is considered one of the most promising approaches for producing pharmaceutical proteins. As has happened with other recombinant protein production schemes, the downstream processing (dsp of these proteins produced in plants is key to the technical and economic success of large-scale applications. Since dsp of proteins produced transgenically in plants has not been extensively studied, it is necessary to broaden the investigation in this field in order to more precisely evaluate the commercial feasibility of this route of expression. In this work, we studied the substitution of an IMAC chromatographic step, described in previous work (Azzoni et al., 2002, with ion-exchange chromatography on SP Sepharose Fast Flow resin as the second step in the purification of recombinant aprotinin from transgenic maize seed. The main goal of this second purification step is to separate the recombinant aprotinin from the native corn trypsin inhibitor. Analysis of the adsorption isotherms determined at 25°C under different conditions allowed selection of 0.020 M Tris pH 8.5 as the adsorption buffer. The cation-exchange chromatographic process produced a high-purity aprotinin that was more than ten times more concentrated than that generated using an IMAC step.

  8. Cardiac and pleuropulmonary AL amyloid imaging with technetium-99m labelled aprotinin

    Energy Technology Data Exchange (ETDEWEB)

    Aprile, C. [Dept. of Nuclear Medicine, Fondazione Clinica del Lavoro-IRCCS, Pavia (Italy); Marinone, G. [Inst. of Clinical Medicine II and Research Lab. Biotechnology, Policlinico S. Matteo-IRCCS, Pavia (Italy); Saponaro, R. [Dept. of Nuclear Medicine, Fondazione Clinica del Lavoro-IRCCS, Pavia (Italy); Bonino, C. [SORIN Biomedica, Saluggia VC (Italy); Merlini, G. [Inst. of Clinical Medicine II and Research Lab. Biotechnology, Policlinico S. Matteo-IRCCS, Pavia (Italy)

    1995-12-01

    Antiproteases are known to be present in amyloid deposits. We evaluated the possibility of using an anti-serine protease (aprotinin) labelled with technetium-99m (TcA), usually employed as a cortical renal tracer, for the imaging of amyloid deposits. Because of the known high uptake of TcA by the kidneys, we limited our analysis to extra-abdominal amyloid localizations. We report the scintigraphic findings observed in 24 patients with light chain amyloidosis (AL) and one with a hereditary form who were known or suspected to have extra-abdominal involvement. Planar scans obtained 100 min after i.v. TcA administration showed myocardial accumulation in 11 patients, pleuropulmonary accumulation in nine, pericardial accumulation in two and localization in the neck region (thyroid, salivary glands and tongue) in eight. TcA scintigraphy was negative in five patients without clinical or laboratory evidence of extra-abdominal involvement, as well as in 12 control group patients with cardiac and renal diseases. These preliminary results indicate TcA to be a low-cost, readily available radiopharmaceutical for imaging of extra-abdominal involvement in AL type amyloidosis. (orig.)

  9. /sup 99m/Tc-aprotinin: A new tracer for kidney morphology and function

    Energy Technology Data Exchange (ETDEWEB)

    Bianchi, C.; Donadio, C.; Tramonti, G.; Lorusso, P.; Bellitto, L.; Lunghi, F.

    1984-01-01

    Aprotinin (Ap), a low molecular weight polyeptide (6500 dalton), is a protease inhibitor which is electively and stably accumulated in the kidney. In 112 adult patients, with either uni- or bilateral renal disease with different degrees of renal impairment (from normal GFR to advanced renal failure), renal scans were performed by means of Ap labelled with /sup 99m/Tc. Highly satisfactory renal scans were obtained in all patients. In 20 patients with renal failure (serum creatinine 1.8 - 8.5 mg/dl, mean 4.7) a comparison was made of the renal scans obtained with /sup 99m/Tc-Ap and with /sup 99m/Tc-DMSA. /sup 99m/Tc-Ap was slightly better than /sup 99m/Tc-DMSA, especially in patients with far advanced renal failure. Some aspects of the pharmacokinetics of /sup 99m/Tc-Ap were studied in 72 cases. In 22 of these patients plasma clearance of /sup 99m/Tc-Ap was determined by the single injection method using a two-compartment model. In patients with GFR>90 ml/min plasma cl of /sup 99m/Tc-Ap was 67.6 +- 8.4 SD ml/min. A good correlation was observed between plasma clearance of /sup 99m/Tc-Ap and GFR (r = 0.74). After i.v. injection /sup 99m/Tc-Ap was stably fixed by the kidney. Renal radioactivity remained stable between the 2nd and the 8th hour after the injection. Urinary excretion of radioactivity measured in 35 patients in the first and in the second 2-hour interval after i.v. injection of /sup 99m/Tc-Ap was negligible in all patients (2.7 +- 1.5 SD percent of the dose in the fist 2 hours; 2.8 +- 1.4 SD between the 2nd and the 4th hour). Conclusions. /sup 99m/Tc-Ap is an excellent agent for renal imaging. It also seems promising for renal function studies.

  10. Protective effects of the antioxidant Ginkgo biloba extract and the protease inhibitor aprotinin against Leiurus quinquestriatus venom-induced tissue damage in rats

    Directory of Open Access Journals (Sweden)

    A. J. Fatani

    2006-04-01

    Full Text Available Oxidative stress and proteases have been implicated in several diseases and extensive evidence indicates that antioxidants and protease inhibitors help prevent organ functional damage. Leiurus quinquestriatus (LQQ scorpion venom causes cellular injuries that may lead to multiple organ failure. Thus, the capability of the antioxidant "natural standardized extract of Gingko biloba leaves (Gin, EGb 761" and the non-selective protease inhibitor, aprotinin, in ameliorating venom-induced biochemical alterations indicative of cellular injury and oxidative stress was studied to determine their effectiveness in protecting rats from venom-evoked cellular damages. Thus, in this study, rats were treated with LQQ venom (0.3mg.kg-1, subcutaneously alone or after Gin (150mg.kg-1, orally, daily for 2 weeks before venom and/or aprotinin (Apr, 46000 KIU.kg-1, intraperitoneally, 30 min before venom. Control groups were injected with saline or treatment modalities. Lungs and hearts were excised after decapitating rats (n=8/group 60 min after venom injection and the following activities were measured: reduced glutathione (GSH, malondialdehyde (MDA - an index of lipid peroxidation, glutathione peroxidase (GPx, glucose-6-phosphate dehydrogenase (G6PD, and lactate dehydrogenase (LDH. Our findings demonstrate that LQQ venomsignificantly elevated GSH (p<0.05 vs. control, MDA (p<0.05, G6PD (p<0.05, and LDH activities (p<0.001 in hearts of envenomed rats. The venom also elevated MDA (p<0.05 vs. control and reduced GSH and GPx (p<0.05 in the lungs of envenomed rats. In general, pretreatment with EGb761 attenuated LQQ venom-evoked increases in GSH (p<0.05 vs. venom, MDA in rat hearts and lungs (p<0.05 vs. venom, plus LDH in the heart (p<0.01. Aprotinin alone significantly reduced the venom-elicited increase in G6PD and LDH activities and the decrease in GPx levels (p<0.05. In general, these protective effects of EGb761 on GSH, MDA (p<0.01 vs. venom and LDH (p<0.001 in the

  11. Estudo comparativo do emprego da aprotinina em baixas doses X placebo, durante a circulação extracorpórea Comparative study of low-dose aprotinin x placebo during cardiopulmonary bypass

    Directory of Open Access Journals (Sweden)

    José Carlos D. V. PONTES

    2002-03-01

    Full Text Available FUNDAMENTOS: A aprotinina, um antifibrinolítico de natureza protéica, tem sido proposta pela literatura, no intuito de minimizar os efeitos adversos da circulação extracorpórea ao sistema fibrinolítico, permitindo assim uma hemostasia mais adequada. OBJETIVO: Estudar comparativamente o efeito da utilização profilática de baixas doses de aprotinina em pacientes submetidos à circulação extracorpórea. MÉTODO: Dezessete pacientes portadores de valvopatia mitral submetidos a correção cirúrgica utilizando-se circulação extracorpórea foram aleatoriamente divididos em dois grupos: I (controle -- 9 pacientes nos quais foi administrado placebo no perfusato e a cada hora de CEC; II (aprotinina -- 8 pacientes nos quais foram administrados 30000 KIU/kg e 7500 KIU/kg de aprotinina adicionados ao volume do perfusato a cada hora de CEC. O volume total de sangramento pós-operatório nas primeiras 24 horas foi monitorado, assim como amostras sangüíneas foram colhidas após indução anestésica e após a neutralização da heparina com sulfato de protamina para que os seguintes parâmetros fossem avaliados: atividade de protrombina (AP, tempo de tromboplastina parcial ativado (TTPA, tempo de trombina (TT, dosagem de fibrinogênio, concentração de antitrombina III (ATIII, tempo de lise da euglobulina (TLE e dosagem do dímero --D (DDi. RESULTADOS: O volume médio total de sangramento nas primeiras 24 horas no grupo controle foi 690,67 ± 377,12, enquanto que no grupo da aprotinina em baixas doses foi de 248,75 ± 103,13 (p=0,0017. No quadro abaixo estão apresentados os resultados obtidos a partir das análises das amostras sangüíneas pré e pós-operatórias comparativas dos grupos controle (I e aprotinina (II. CONCLUSÃO: A utilização da aprotinina, em baixa dose, foi eficaz na redução do sangramento, assim na diminuição da fibrinólise.BACKGROUND: The use of aprotinin, a antifibrinolytic agent, has been shown to decrease

  12. Aprotinina preserva plaquetas em crianças com cardiopatia congênita acianogênica operadas com circulação extracorpórea? Does aprotinin preserve platelets in children with acyanogenic congenital heart disease undergone surgery with cardiopulmonary bypass?

    Directory of Open Access Journals (Sweden)

    Cesar Augusto Ferreira

    2009-09-01

    Full Text Available OBJETIVO: Avaliação dos efeitos hemostáticos e plaquetários em crianças submetidas a correção de cardiopatias congênitas acianogênicas com circulação extracorpórea que receberam aprotinina. MÉTODOS: Estudo prospectivo randomizado em crianças de 30 dias a 4 anos de idade, submetidas a correção de cardiopatia congênita acianogênica, com circulação extracorpórea (CEC e divididas em dois grupos, um denominado Controle (n=9 e o outro, Aprotinina (n=10. Neste, a droga foi administrada antes e durante a CEC. A disfunção hemostática foi analisada por marcadores clínicos e bioquímicos. Foram consideradas significantes as diferenças com POBJECTIVE: Evaluation of the hemostatic and platelets effects in children with acyanogenic congenital heart disease undergone on-pump surgery who received aprotinin. METHODS: A prospective randomized study was performed on children aged 30 days to 4 years who had undergone correction of acyanogenic congenital heart disease using cardiopulmonary bypass (CPB and were divided into two groups: Control (n=9 and Aprotinin (n=10. In the Aprotinin Group the drug was administered before and during CPB and the hemostatic dysfunction was analyzed by clinical and biochemical markers. Differences were considered to be significant when P<0.05. RESULTS: The groups were similar regarding demographic and intraoperative variables, except for a greater hemodilution in the Aprotinin Group. The drug presented no benefit regarding time of mechanical pulmonary ventilation, stay in the postoperative intensive care unit and hospital, or regarding the use of inotropic drugs and renal function. Platelet concentration was preserved with the use of Aprotinin, whereas thrombocytopenia occurred in the Control Group since the initiation of CPB. Blood loss was similar for both groups. There were no complications with the use of Aprotinin. CONCLUSION: Aprotinin quantitatively preserved the blood platelets in children with

  13. Avaliação da aprotinina na redução da resposta inflamatória sistêmica em crianças operadas com circulação extracorpórea Assessment of aprotinin in the reduction of inflammatory systemic response in children undergoing surgery with cardiopulmonary bypass

    Directory of Open Access Journals (Sweden)

    Cesar Augusto Ferreira

    2010-03-01

    Full Text Available OBJETIVO: Avaliar se a aprotinina em altas doses hemostáticas pode reduzir o processo inflamatório após circulação extracorpórea (CEC em crianças. MÉTODOS: Estudo prospectivo randomizado em crianças de 30 dias a 4 anos de idade, submetidas à correção de cardiopatia congênita acianogênica, com CEC e divididas em dois grupos, um denominado Controle (n=9 e o outro, Aprotinina (n=10. Neste, o fármaco foi administrado antes e durante a CEC. A resposta inflamatória sistêmica e disfunções hemostática e multiorgânicas foram analisadas por marcadores clínicos e bioquímicos. Foram consideradas significantes as diferenças com POBJECTIVE: To assess if the hemostatic high-dose aprotinin is able to reduce the inflammatory process after cardiopulmonary bypass (CPB in children. METHODS: A prospective randomized study was performed on children aged 30 days to 4 years who underwent correction of acyanogenic congenital heart disease with CPB and were divided into two groups: Control (n=9 and Aprotinin (n=10. In the Aprotinin Group the drug was administered before and during CPB and the systemic inflammatory response and hemostatic and multiorgan dysfunctions were assessed through clinical and biochemical markers. Differences were considered to be significant when P<0.05. RESULTS: The groups were similar regarding demographic and intraoperative variables, except for a greater hemodilution in the Aprotinin Group. The drug had no benefit regarding time of mechanical pulmonary ventilation, staying in the postoperative ICU and length of hospitalization, or regarding the use of inotropic drugs and renal function. The partial arterial oxygen pressure/ inspired oxygen fraction ratio (PaO2/FiO2 was significantly reduced 24 h after surgery in the Control Group. Blood loss was similar for both groups. Significant leukopenia was observed in the Aprotinin Group during CPB, followed by leukocytosis. Tumor necrosis factor alpha (TNF- α, interleukins (IL

  14. Anestesia para tratamento de aspergilose cardíaca em paciente com trombocitopenia: o uso criterioso da aprotinina Anestesia para tratamiento de aspergilosis cardiaca en paciente con trombocitopenia: el uso con criterio de la aprotinina Anesthesia for treatment of cardiac aspergillosis in a patient with thrombocytopenia and the judicious use of aprotinin

    Directory of Open Access Journals (Sweden)

    Raquel Reis Soares

    2007-12-01

    : Aprotinin has been widely used in cardiac surgeries as a therapeutic resource for reducing the effects of cardiopulmonary bypass (CPB on coagulation and fibrinolysis. Recovery of adequate hemostasia at the end of the procedure is one of the objectives of the anesthesiologist. However, aprotinin has specific indications. The objective of this report was to present the case of a patient with severe thrombocytopenia undergoing cardiac surgery in which consultation with Hematology and adequate planning were responsible for the success of the procedure. CASE REPORT: An 18-year old male patient, weighing 64 kg, physical status ASA IV, with a diagnosis of bone marrow aplasia, was being investigated to undergo bone marrow transplantation. He had persistent fever for a month, which did not improve with antibiotics. During the investigation with imaging exams, a left atrial mass was discovered. Laboratory exams revealed hemoglobin 9 g.dL-1 and thrombocytopenia with 6,000 platelets.mm³. He underwent a sternotomy with CPB to remove the intracavitary thrombus. In order to control intraoperative bleeding, the following was administered: plateletpheresis, hydrocortisone, and aprotinin. Increased bleeding and hemodynamic instability did not develop during the surgery, and the patient was transferred to the Intensive Care Unit (ICU without intercurrences. The anatomo-pathologic exam revealed the thrombus to be filled with Aspergillus (fungal mass. On the seventh postoperative day the patient developed respiratory failure and cardiorespiratory arrest that did not respond to resuscitation maneuvers. CONCLUSIONS: Despite the increased risk of bleeding in this patient, cardiac surgery with CPB was performed without intercurrences due to the use of aprotinin and plateletpheresis.

  15. 抑肽酶在体外循环肺保护机制中的作用%Protective Effect of Aprotinin on Lungs in Cardiopulmonary Bypass Patients

    Institute of Scientific and Technical Information of China (English)

    魏磊; 刘标; 梁永年; 陈亦江

    2004-01-01

    目的研究CPB的肺损伤和抑肽酶的肺保护作用.方法选择24例患者随机分为两组(各12例),实验组在体外循环机中加入抑肽酶10万KIU/Kg;对照组则不用抑肽酶.动态检测两组患者左右房中性粒细胞(PMN)、血小板(Pt)、IL-6、IL-10、TNF-α、肺泡氧合指数(OI).结果两组转流前左、右房PMN、Pt无显著差异,对照组主动脉开放心脏复跳后5分钟检测左房PMN、Pt明显低于右房(P<0.05);实验组左、右心房血中PMN、Pt无明显差异(P>0.05).CPB开始后,两组桡动脉血IL-6、IL-10、TNF-α进行性增高,对照组IL-6、TNF-α增高更显著(P<0.05).实验组IL-10增高更显著(P<0.05).CPB后实验组患者肺泡氧合指数(OI)明显低于对照组(P<0.05).结论抑肽酶能通过保护血小板,抑制炎症因子IL-6、TNF-α的产生,并能加速抗炎因子IL-10的释放,从而抑制白细胞的激活、肺内聚集和扣留,减轻体外循环后肺损伤和通气功能障碍,改善术后肺功能.

  16. 抑肽酶对血小板膜糖蛋白功能的影响%Effects of aprotinin on platelet membrane glycoprotein function

    Institute of Scientific and Technical Information of China (English)

    仲焰; 隋波; 尹格平; 李训美

    2001-01-01

    目的探讨抑肽酶对心肺转流(CPB)期间血小板功能的保护作用.方法选择20例先天性心脏病房、室间隔缺损在CPB下行心内修补术的病人,随机分为对照组(不加抑肽酶)和抑肽酶组(加抑肽酶200万KIU),每组10例.分别测定不同时间点的血小板计数和血小板膜糖蛋白GPI b、GPⅡb-Ⅲa的表达.结果与对照组相比,抑肽酶组血小板计数和血小板膜糖蛋白GPI b、GPⅡb-Ⅲa的表达减少明显减轻(P<0.05),术后出血也明显减少.结论抑肽酶在体外循环期间可抑制血小板计数和血小板膜糖蛋白GPI b和GPⅡb-Ⅲa的减少,对血小板功能具有保护作用.

  17. High-Dose Aprotinin in Cardiac Surgery:Is High-Dose High Enough?An Analysis of 8281 Cardiac Surgical Patients Treated with Aprotinin%"大剂量"抑肽酶剂量足够大吗?8281例心脏手术患者应用大剂量抑肽酶的临床分析

    Institute of Scientific and Technical Information of China (English)

    Wulf Dietrich; Raimund Busley; Monika Kriner; 周仁龙

    2007-01-01

    有假说认为抑肽酶剂量大于6×106 KIU(kallikrein inhibiting units,激肽释放酶抑制单位)时对减少出血的效果优于(5~6)×106 KIU的大剂量方案,在本回顾性分析中,我们对此进行了验证.8281例成年心脏手术患者根据体重公斤数与手术时间分钟数给予抑肽酶.采用线性分析与logistic回归模型,以检测抑肽酶剂量与手术后出血、输血及其他因素之间的相关性.最低四分位数剂量组与最高四分位数剂量组的手术后6小时胸导管引流量分别为447±319 ml和360±290 ml(p<0.001),需异体血输注的比例分别为55%和47%(P<0.01).抑肽酶剂量可以作为需再次开胸手术止血的一个独立性的预测因素(最高四分位数组为1.9%,最低四分位数组为2.4%,P<0.01).大剂量组肾衰竭需透析的风险(大剂量组为2.3%,小剂量组为3.3%,P<0.01)及出现肾功能受损(手术后肌酐水平升高≥2 mg/dl,大剂量组为6.4%,小剂量组为10.0%,P<0.01)的风险较低.因此,抑肽酶剂量和肾功能之间无明显相关性.我们的结果支持在心脏手术中应用更大、更个体化剂量的抑肽酶可能会取得更好的治疗效果.

  18. 不同剂型药用抑肽酶纯度的胶束电动毛细管色谱测定%Determination of Purity of Different Types of Aprotinin by Micellar Electrokinetic Capillary Chromatography

    Institute of Scientific and Technical Information of China (English)

    姜廷福; 陆豪杰; 李辰; 梁冰; 欧庆瑜

    2002-01-01

    以十六烷基三甲基溴化铵(CTAB)为阳离子表面活性剂,用胶束电动毛细管色谱(MECC)分别对抑肽酶粉针剂和抑肽酶注射液进行纯度测定.实验中选择了最佳缓冲液(含4 mmol/L CTAB的80 mmol/L Na2HPO4-H3PO4,pH 7.00),考察了进样量与样品中高浓度盐对分离的影响.并对毛细管区带电泳、MECC和高效液相色谱的分离效果加以比较,表明MECC的分离效果最佳.

  19. Urinary epidermal growth factor is excreted from the rat isolated perfused kidney in the absence of plasma

    DEFF Research Database (Denmark)

    Jørgensen, P E; Hilchey, S D; Nexø, Ebba;

    1993-01-01

    . Administration of the proteinase inhibitor aprotinin reduced urinary EGF excretion from the rat isolated perfused kidney by approximately 50%. In conclusion, the rat isolated perfused kidney excreted significant amounts of urinary EGF without having access to plasma, and EGF excretion was reduced by aprotinin...

  20. [Anaphylactic shock caused by fibrin glue].

    Science.gov (United States)

    Orsel, I; Guillaume, A; Feiss, P

    1997-01-01

    We observed a case of anaphylactic shock in a 68-year-old woman after a nasal intramucosal injection of fibrin glue for telangiectasies therapy. The tests showed an allergy to aprotinin contained in the glue. In the previous years, glue and aprotinin had been administered to the patient several times for nasal bleeding.

  1. Adaption of Saccharomyces cerevisiae expressing a heterologous protein

    DEFF Research Database (Denmark)

    Krogh, Astrid Mørkeberg; Beck, Vibe; Højlund Christensen, Lars;

    2008-01-01

    Production of the heterologous protein, bovine aprotinin, in Saccharomyces cerevisiae was shown to affect the metabolism of the host cell to various extent depending on the strain genotype. Strains with different genotypes, industrial and laboroatory, respectively, were investigated. The maximal...

  2. Alternativas farmacológicas a la transfusión sanguínea: ¿Qué hay de nuevo?

    OpenAIRE

    2004-01-01

    Pharmacological approaches to reduce blood transfusion include the protease inhibitor aprotinin, lysine-analogue antifibrinolytics synthetic arginine-vasopressin derivatives (DDAVP) and recombinant factor VII (rfVIIa). These agents are known to prevent the need for blood after major surgery (cardiac, hepatic, and orthopaedic). Among the nonhemostatic agents erythropoietin (EPO) may be effective to reduce blood requirements in medical and surgical patients. Aprotinin is consistently effective ...

  3. Aprotininskintigrafi ved amyloidose

    DEFF Research Database (Denmark)

    Haraldsen, Ate; Eskild-Jensen, Anni; Frøkiær, Jørgen

    2009-01-01

    Amyloidosis is a disease characterized by protein deposition (amyloid) in THE extracellular matrix leading to organ dysfunction and death. New treatment modalities have emphasized the need for accurate and early diagnosis. Aprotinin scintigraphy is a radionuclide imaging technique in which...... the localisation and extent of amyloid deposits are visualized. It is specific and sensitive in detecting cardiac amyloidosis, which is associated with a poor prognosis. In addition, aprotinin scintigraphy appears to be a useful tool for the monitoring of disease progression and treatment efficacy. Udgivelsesdato...

  4. Identification of fibrin clot-bound plasma proteins

    NARCIS (Netherlands)

    S. Talens (Simone); F.W.G. Leebeek (Frank); J.A.A. Demmers (Jeroen); D.C. Rijken (Dingeman)

    2012-01-01

    textabstractSeveral proteins are known to bind to a fibrin network and to change clot properties or function. In this study we aimed to get an overview of fibrin clot-bound plasma proteins. A plasma clot was formed by adding thrombin, CaCl2 and aprotinin to citrated platelet-poor plasma and unbound

  5. The influence of retention on the plate height in ion-exchange chromatography

    DEFF Research Database (Denmark)

    Hansen, Ernst; Mollerup, Jørgen

    2004-01-01

    The plate heights for the amino acid tyrosine (anion exchange) and the polypeptide aprotinin (cation exchange) were determined on a porous media (Resource 15) and a get filled media (HyperD 20) at salt concentrations ranging from weak to strong retention. At a constant velocity, measurements show...

  6. Prevention of Bleeding in Orthognathic Surgery--A Systematic Review and Meta-Analysis of Randomized Controlled Trials

    DEFF Research Database (Denmark)

    Olsen, Jesper J; Skov, Jane; Ingerslev, Janne

    2016-01-01

    and operating time. This review is registered at PROSPERO (CRD42014014840). RESULTS: Eleven trials were included for review. The individual trials demonstrated the effects on IOB from hypotensive anesthetic regimens, the use of aprotinin, and the herbal medicine Yunnan Baiyao. Six studies of tranexamic acid...

  7. Thrombocyte and thrombocyte function

    Institute of Scientific and Technical Information of China (English)

    1997-01-01

    970384 The mechanism of preserving platelet func-tion by aprotinin during cardiopulmonary bypass.YANG Tianyu(杨天宇), et al. Anesthesiol &Cardiòpulmon Bypass Lab, Cardiovasc Instit & FuwaiHosp, CAMS & PUMC, Beijing, 100037. Chin Cir J1996; 11(12): 739-742.

  8. Stability of glucagon-like peptide-1 and glucagon in human plasma

    DEFF Research Database (Denmark)

    Albrechtsen, Nicolai Jacob Wewer; Bak, Monika Judyta; Hartmann, Bolette;

    2015-01-01

    To investigate the stability of glucagon-like peptide-1(GLP-1) and glucagon in plasma under short- and long-term storage conditions. Methods: Pooled human plasma (n=20), to which a dipeptidyl peptidase 4 (DPP-4) inhibitor and aprotinin were added, was spiked with synthetic GLP-1 (intact, 7-36NH2 ...

  9. Drug: D08813 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available tic category of drugs in Japan [BR:br08301] 7 Agents not mainly for therapeutic purpose 79 Other agents not ...mainly for therapeutic purpose 799 Miscellaneous 7990 Miscellaneous D08813 Aprotinin - thrombin - human fibrinogen mixt PubChem: 96025496 ...

  10. Antifibrinolytics in liver surgery

    Directory of Open Access Journals (Sweden)

    Jalpa Makwana

    2010-01-01

    Full Text Available Hyperfibrinolysis, a known complication of liver surgery and orthotopic liver transplantation (OLT, plays a significant role in blood loss. This fact justifies the use of antifibrinolytic drugs during these procedures. Two groups of drug namely lysine analogues [epsilon aminocaproic acid (EACA and tranexamic acid (TA] and serine-protease-inhibitors (aprotinin are frequently used for this purpose. But uniform data or guidelines on the type of antifibrinolytic drugs to be used, their indications and correct dose, is still insufficient. Antifibrinolytics behave like a double-edged sword. On one hand, there are benefits of less transfusion requirements but on the other hand there is potential complication like thromboembolism, which has been reported in several studies. We performed a systematic search in PubMed and Cochrane Library, and we included studies wherein antifibrinolytic drugs (EACA, TA, or aprotinin were compared with each other or with controls/placebo. We analysed factors like intraoperative red blood cell and fresh frozen plasma requirements, the perioperative incidence of hepatic artery thrombosis, venous thromboembolic events and mortality. Among the three drugs, EACA is least studied. Use of extensively studied drug like aprotinin has been restricted because of its side effects. Haemostatic effect of aprotinin and tranexamic acid has been comparable. However, proper patient selection and individualized treatment for each of them is required. Purpose of this review is to study various clinical trials on antifibrinolytic drugs and address the related issues like benefits claimed and associated potential complications.

  11. Efficacy and Safety of Antifibrinolytic Agents in Reducing Perioperative Blood Loss and Transfusion Requirements in Scoliosis Surgery: A Systematic Review and Meta-Analysis

    OpenAIRE

    Meng Wang; Xin-Feng Zheng; Lei-Sheng Jiang

    2015-01-01

    Background Routine use of antifibrinolytic agents in spine surgery is still an issue of debate. Objective To gather scientific evidence for the efficacy and safety of antifibrinolytic agents including aprotinin, tranexamic acid (TXA) and epsilon aminocaproic acid (EACA, traditionally known as Amicar) in reducing perioperative blood loss and transfusion requirements in scoliosis surgery. Methods We conducted a systematic review and meta-analysis for randomized controlled trials (RCTs), retrosp...

  12. Identification of proteolytic activities in ROS 17/2.8 cell lysates which cleave peptide substrates for protein kinase C-mediated phosphorylation.

    Science.gov (United States)

    Guidon, P T; Harrison, P

    1996-04-01

    We have observed two proteolytic activities in cell lysates from the rat osteoblastic osteosarcoma cell line ROS 17/2.8 which are capable of cleaving a peptide substrate for protein kinase C-mediated phosphorylation, and other peptides containing similar sequences. Both activities are inhibited by Pefabloc, a serine protease inhibitor, while one of the activities is inhibited by either EDTA or aprotinin. The protease inhibitors pepstatin, bestatin, E-64, leupeptin and phosphoramidon do not block either of these proteolytic activities.

  13. Cell Growth Arrest Mediated by STAT Proteins in Breast Cancer Cells

    Science.gov (United States)

    1998-07-01

    pepstatin, and aprotinin (1 (Xg/ml each). Whole cell extracts were immediately subjected to electromobility shift assay. Preparation of membrane and...activation on the cytosol fraction (STAT protein) concentration. Electromobility shift assay (EMSA) The sample after in vitro activation (3 ul) (1 [il...transcription; EGF, epidermal growth factor; NGF, nerve growth factor; EMSA, electromobility shift assay; SIF, sis-inducible factor; SIE, sis

  14. Pharmacological manipulation of gastric juice: thrombelastographic assessment and implications for treatment of gastrointestinal haemorrhage.

    Science.gov (United States)

    Patchett, S E; O'Donoghue, D P

    1995-03-01

    The impairment of formation and maintenance of a formed fibrin clot contributes to the prolonged bleeding and high incidence of rebleeding in upper gastrointestinal haemorrhage. To investigate the basis for the use of drug therapy in gastric bleeding, this study used thrombelastography to determine the effects of pharmacological manipulation of gastric juice on coagulation and fibrinolysis. The thrombelastograph is a mechanical device that provides a visual assessment of all stages of coagulation and fibrinolysis. The effects of fresh and pharmacologically changed gastric juice was assessed after its addition to fresh whole blood in the thrombelastograph cuvette. Pharmacological manipulation was achieved through alkalisation or through addition of tranexamic acid, aprotinin, or sucralfate. Fresh gastric juice delayed clot formation, decreased maximum clot amplitude, and stimulated clot lysis. Alkalisation inhibited the lytic effects of fresh gastric juice and improved the induced abnormalities in coagulation. Tranexamic acid partially inhibited gastric juice induced clot lysis but did not exhibit a beneficial effect on coagulation. Sucralfate, and to a lesser extent aprotinin significantly inhibited fibrinolysis but exacerbated the detrimental effect of gastric juice on the parameters of coagulation. Alkalisation of gastric juice reduces the adverse effect on coagulation and fibrinolysis. Tranexamic acid, aprotinin, and sucralfate can all reduce or inhibit clot lysis, but the adverse effects on clot formation may outweigh any potential benefit in the treatment of gastrointestinal bleeding.

  15. Influence of storage conditions on in vitro stability of atrial natriuretic peptide and of anesthesia on plasma atrial natriuretic peptide concentration in cats.

    Science.gov (United States)

    Heishima, Yasuhiro; Hori, Yasutomo; Chikazawa, Seishiro; Kanai, Kazutaka; Hoshi, Fumio; Itoh, Naoyuki

    2016-08-01

    OBJECTIVE To investigate the in vitro stability of atrial natriuretic peptide (ANP) in plasma samples under various storage conditions and the influence of anesthesia on plasma ANP concentration in cats. ANIMALS 1 cat with congestive heart failure and 5 healthy adult mixed-breed cats. PROCEDURES A plasma sample from the cat with heart failure was serially diluted, and dilutional parallelism of ANP concentration was evaluated. Plasma samples containing aprotinin or serum samples from the 5 healthy cats were kept at room temperature (27°C) for ≤ 12 hours. Plasma samples from the same healthy cats were stored at -70°, -20°, or 4°C for ≤ 14 days. Plasma samples were obtained from the healthy cats before and during isoflurane anesthesia. Plasma ANP concentrations were measured at a commercial laboratory by use of a human ANP chemiluminescence assay. RESULTS Intra- and interassay coefficients of variation were 1.5% and 2.5%, respectively, and dilutional parallelism was established. Although ANP concentration decreased by 82.4 ± 13.6% (mean ± SD) after sample storage for 12 hours at room temperature, this decrease was prevented by aprotinin. Plasma ANP concentrations were stable for 7 days at -20°C and for 14 days at -70°C. However, concentrations decreased markedly to 57.6 ± 6.9% at -20°C and to 18.0 ± 3.0% at 4°C after 14 days. Plasma ANP concentration decreased significantly in cats during anesthesia and was correlated with blood pressure. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that aprotinin should be added routinely in preparation of plasma samples from cats for measurement of ANP concentration, and those samples, if stored, should be frozen immediately at ≤ -20°C. General anesthesia or systemic blood pressure may affect plasma ANP concentration in cats.

  16. Divergent Inhibitor Susceptibility among Airway Lumen-Accessible Tryptic Proteases.

    Directory of Open Access Journals (Sweden)

    Shilpa Nimishakavi

    Full Text Available Tryptic serine proteases of bronchial epithelium regulate ion flux, barrier integrity, and allergic inflammation. Inhibition of some of these proteases is a strategy to improve mucociliary function in cystic fibrosis and asthmatic inflammation. Several inhibitors have been tested in pre-clinical animal models and humans. We hypothesized that these inhibitors inactivate a variety of airway protease targets, potentially with bystander effects. To establish relative potencies and modes of action, we compared inactivation of human prostasin, matriptase, airway trypsin-like protease (HAT, and β-tryptase by nafamostat, camostat, bis(5-amidino-2-benzimidazolylmethane (BABIM, aprotinin, and benzamidine. Nafamostat achieved complete, nearly stoichiometric and very slowly reversible inhibition of matriptase and tryptase, but inhibited prostasin less potently and was weakest versus HAT. The IC50 of nafamostat's leaving group, 6-amidino-2-naphthol, was >104-fold higher than that of nafamostat itself, consistent with suicide rather than product inhibition as mechanisms of prolonged inactivation. Stoichiometric release of 6-amidino-2-naphthol allowed highly sensitive fluorometric estimation of active-site concentration in preparations of matriptase and tryptase. Camostat inactivated all enzymes but was less potent overall and weakest towards matriptase, which, however was strongly inhibited by BABIM. Aprotinin exhibited nearly stoichiometric inhibition of prostasin and matriptase, but was much weaker towards HAT and was completely ineffective versus tryptase. Benzamidine was universally weak. Thus, each inhibitor profile was distinct. Nafamostat, camostat and aprotinin markedly reduced tryptic activity on the apical surface of cystic fibrosis airway epithelial monolayers, suggesting prostasin as the major source of such activity and supporting strategies targeting prostasin for inactivation.

  17. Suppression of BRCA2 by Mutant Mitochondrial DNA in Prostate Cancer

    Science.gov (United States)

    2011-05-01

    Nonidet P - 40 , 2 mmol/L phenylmethylsulfonyl fluoride, 10 g/mL aprotinin, 10 g/mL leupeptin, 10 mmol/L sodium fluoride, 1 mmol/L sodium orthovanadate, and...specimens were pulverized and homogenized in lysis buffer containing 0.1% SDS, 1% Non- idet P - 40 , 50 mmol/L Tris-HCl (pH 7.5), 150 mmol/L NaCl, 200...Analyze the correlation between type/number of mtDNA mutations and clinical stage (Months 10-11) We extracted total DNA from 6 BPH and 40 PCA frozen

  18. Yin and Yang of Heparanase in Breast Tumor Initiation

    Science.gov (United States)

    2013-04-01

    Nonidet P (NP)- 40 lysis buffer (50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1% NP- 40 , 5 mM EDTA, 10 µg/ml aprotinin, 10 µg/ml leupeptinin, and 1 mM...532- 40 . 28. Xu X, Quiros RM, Gattuso P , Ain KB, and Prinz RA High prevalence of BRAF gene mutation in papillary thyroid carcinomas and thyroid tumor...enzymatic activity. Neu, mice enco brea trans TVA sugg Fig. form old) 8C (red each palp and ( p ɘ Fi RC an lev lin

  19. The membrane fraction of homogenized rat kidney contains an enzyme that releases epidermal growth factor from the kidney membranes

    DEFF Research Database (Denmark)

    Nexø, Ebba; Poulsen, Steen Seier

    1991-01-01

    High levels of epidermal growth factor (EGF) are excreted in the urine and high levels of mRNA for the EGF-precursor have been demonstrated in the kidney. The EGF-precursor is a membrane bound peptide in the kidney, but little is known about the renal processing of the precursor. The present stud....... The EGF releasing enzyme is inhibited by the serine proteinase inhibitor aprotinin and by low temperatures (4 degrees C). The pH optimum of the reaction is pH 7.5-8.0....

  20. Plasminogen-independent initiation of the pro-urokinase activation cascade in vivo. Activation of pro-urokinase by glandular kallikrein (mGK-6) in plasminogen-deficient mice

    DEFF Research Database (Denmark)

    List, K; Jensen, Ole Nørregaard; Bugge, T H;

    2000-01-01

    kallikrein). The pro-uPA converting activity of the mGK-6 enzyme, as well as its ability to cleave a synthetic substrate for glandular kallikrein, was inhibited by the serine proteinase inhibitor leupeptin but not by other serine proteinase inhibitors such as aprotinin, antithrombin III, or alpha(1...... the cascade by activating pro-uPA. The urine from Plg -/- mice contained active two-chain uPA as well as a proteinase capable of activating exogenously added pro-uPA. The active component was purified and identified by mass spectrometry-based peptide mapping as mouse glandular kallikrein mGK-6 (true tissue...

  1. Plasminogen-independent initiation of the pro-urokinase activation cascade in vivo. Activation of pro-urokinase by glandular kallikrein (mGK-6) in plasminogen-deficient mice

    DEFF Research Database (Denmark)

    List, K; Jensen, O N; Bugge, T H;

    2000-01-01

    , and as for other cascade systems, understanding the physiological initiation mechanism is of central importance. The attempts to identify initiation routes for activation of the proform of the key enzyme urokinase-type plasminogen activator (pro-uPA) in vivo have been hampered by the strong activator potency...... kallikrein). The pro-uPA converting activity of the mGK-6 enzyme, as well as its ability to cleave a synthetic substrate for glandular kallikrein, was inhibited by the serine proteinase inhibitor leupeptin but not by other serine proteinase inhibitors such as aprotinin, antithrombin III, or alpha(1...

  2. Hyaluronan-Based Therapy for Metastatic Prostate Cancer

    Science.gov (United States)

    2015-07-01

    cancer cell lines (PC3, DU145) correlate with greater tumorigenic and metastatic properties over prostate cancer cell lines (LNCaP) that do not...tumorigenic and metastatic properties (right). B) Cy5.5-labeled HA-NPs (50 µg/mL) or HA polymer (234.4 kDa) were incubated for 2 hours with PC3 cells. C...lysed with DISC IP lysis buffer (30 mM Tris, pH 7.4, 150 mM NaCl, 10% glycerol , 1% Triton X-100 with 1 mM PMSF, and 1 μg/mL each of aprotinin, leupeptin

  3. Contribution of the Kallikrein/Kinin System to the Mediation of ConA-Induced Inflammatory Ascites.

    Science.gov (United States)

    Baintner, Károly

    2016-03-01

    Intraperitoneal administration of concanavalin A (ConA, 25 mg/kg b.w.), a cell-binding plant lectin was used for inducing inflammatory ascites, and potential inhibitors were tested in 1 h and 2.5 h experiments, i.e. still before the major influx of leucocytes. At the end of the experiment the peritoneal fluid was collected and measured. The ConA-induced ascites was significantly (p<0.01) and dose-dependently inhibited by icatibant (HOE-140), a synthetic polypeptide antagonist of bradykinin receptors. Aprotinin, a kallikrein inhibitor protein also had significant (p<0.01), but less marked inhibitory effect. L-NAME, an inhibitor of NO synthesis, and atropine methylnitrate, an anticholinergic compound, were ineffective. It is concluded, that the kallikrein/kinin system contributes to the mediation of the ConA-induced ascites by increasing subperitoneal vascular permeability, independent of the eventual vasodilation produced by NO. It is known, that membrane glycoproteins are aggregated by the tetravalent ConA and the resulting distortion of membrane structure may explain the activation of the labile prekallikrein. Complete inhibition of the ConA-induced ascites could not be achieved by aprotinin or icatibant, which indicates the involvement of additional mediators.

  4. Calcium binding to cardiac myocytes protected from proteolytic enzyme activity.

    Science.gov (United States)

    Bailey, L E; Fawzi, A B

    1985-04-17

    Excitation-contraction coupling in cardiac muscle is dependent on extracellular calcium and calcium bound to the surface of the myocardial cell. In this study, we examined the physical characteristics of calcium binding to adult guinea pig ventricular myocytes disaggregated mechanically in oxygenated tissue culture medium containing a proteinase inhibitor (aprotinin), and separated from cellular debris by Cytodex beads. Cells prepared in this manner excluded Trypan blue and showed no evidence of spontaneous contraction or contracture. Scatchard plots of calcium binding determined by continuous flow equilibrium dialysis revealed a high-affinity, low-capacity pool, Ka = 65 X 10(3) M-1 and Bt = 1.3 nmol X mg-1 and a low-affinity, high-capacity pool, Ka = 141 M-1 and Bt = 138 nmol X mg-1. The low-affinity pool was not detectable after lanthanum, trypsin or collagenase treatment or in cells prepared without aprotinin in the isolation medium. Both neuraminidase and phospholipase C reduced Bt of the low-affinity pool by one half, but only neuraminidase affected the affinity constant of this pool. Ka was increased to 516.7 M-1, similar to the apparent affinity constant for calcium binding estimated from dP/dtmax measured at several extracellular calcium concentrations (470 M-1). The results suggest that calcium bound to sarcolemmal phospholipids represents the superficial calcium involved in excitation-contraction coupling in the heart.

  5. Role of kinin B2 receptors in opioid-induced hyperalgesia in inflammatory pain in mice.

    Science.gov (United States)

    Grastilleur, Sébastien; Mouledous, Lionel; Bedel, Jerome; Etcheverry, Jonathan; Bader, Michael; Girolami, Jean-Pierre; Fourcade, Olivier; Frances, Bernard; Minville, Vincent

    2013-03-01

    Postoperative pain management is a clinical challenge that can be complicated by opioid-induced hyperalgesia (OIH). Kinin receptors could mediate both the acute and chronic phases of inflammation and pain. A few recent studies suggest that dynorphin A could maintain neuropathic pain by activating the bradykinin (BK) receptor. Thus, the effect of a single administration of sufentanil (a μ-opioid receptor agonist) was investigated in a model of carrageenan-induced inflammatory pain using three strains of mice, i.e., knockout mice for one kinin receptor, B1R or B2R (B1KO, B2KO), and wild-type C57/BL6J mice (WT) treated with either a B1R (R954) or a B2R antagonist (HOE140) or a KKS inhibitor (aprotinin). Pain was assessed and compared between the different groups using two behavioral tests exploring mechanical (von Frey filaments) and thermal (Hargreaves test) sensitivity. Pretreatment with sufentanil induced a sustained increase in pain sensitivity with a delayed return to baseline values characterizing an OIH in carrageenan-injected mice only. Sufentanil-induced OIH was not observed in B2KO but persisted in B1KO and was blunted by aprotinin and the B2R antagonist only. Collectively, our data indicate that the B2R receptor and BK synthesis or availability are essential peripheral steps in the mechanism leading to OIH in a pain context.

  6. Crystallization and preliminary X-ray analysis of a Kunitz-type inhibitor, textilinin-1 from Pseudonaja textilis textilis

    Energy Technology Data Exchange (ETDEWEB)

    Millers, Emma-Karin I. [School of Molecular and Microbial Sciences, University of Queensland, Brisbane 4072, QLD (Australia); Masci, Paul P. [School of Medicine, Southern Division, University of Queensland, Princess Alexandra Hospital, Woolloongabba, Brisbane 4102, QLD (Australia); Lavin, Martin F. [The Queensland Cancer Fund Research Unit, The Queensland Institute of Medical Research, PO Box, Royal Brisbane Hospital, Herston, Brisbane 4029, QLD (Australia); Jersey, John de; Guddat, Luke W., E-mail: luke.guddat@uq.edu.au [School of Molecular and Microbial Sciences, University of Queensland, Brisbane 4072, QLD (Australia)

    2006-07-01

    Crystals of a canonical inhibitor of plasmin from Australian Brown snake venom has been obtained. In complex with trypsin these diffract to 2.0 Å resolution, while the free inhibitor diffracts to 1.63 Å. Textilinin-1 (Txln-1), a Kunitz-type serine protease inhibitor, is a 59-amino-acid polypeptide isolated from the venom of the Australian Common Brown snake Pseudonaja textilis textilis. This molecule has been suggested as an alternative to aprotinin, also a Kunitz-type serine protease inhibitor, for use as an anti-bleeding agent in surgical procedures. Txln-1 shares only 47% amino-acid identity to aprotinin; however, six cysteine residues in the two peptides are in conserved locations. It is therefore expected that the overall fold of these molecules is similar but that they have contrasting surface features. Here, the crystallization of recombinant textilinin-1 (rTxln-1) as the free molecule and in complex with bovine trypsin (229 amino acids) is reported. Two organic solvents, phenol and 1,4-butanediol, were used as additives to facilitate the crystallization of free rTxln-1. Crystals of the rTxln-1–bovine trypsin complex diffracted to 2.0 Å resolution, while crystals of free rTxln-1 diffracted to 1.63 Å resolution.

  7. Immobilization of enzymes using non-ionic colloidal liquid aphrons (CLAs): Surface and enzyme effects.

    Science.gov (United States)

    Ward, Keeran; Xi, Jingshu; Stuckey, David C

    2015-12-01

    The use of non-ionic colloidal liquid aphrons (CLAs) as a support for enzyme immobilisation was investigated. Formulation required the mixing of an aqueous-surfactant solution with a relatively non-polar solvent-surfactant solution, forming a solvent droplet surrounded by a thin stabilised aqueous film (soapy shell). Studies utilising anionic surfactants have showed increased retention, however, very little have been understood about the forces governing immobilisation. This study seeks to determine the effects of enzyme properties on CLA immobilisation by examining a non-ionic/non-polar solvent system comprised of two non-ionic surfactants, Tween 20 and 80, mineral oil and the enzymes lipase, aprotinin and α-chymotrypsin. From these results it was deduced that hydrophobic interactions strongly governed immobilisation. Confocal Scanning Laser Microscopy (CSLM) revealed that immobilisation was predominantly achieved by surface adsorption attributed to hydrophobic interactions between the enzyme and the CLA surface. Enzyme surface affinity was found to increase when added directly to the formulation (pre-manufacture addition), as opposed to the bulk continuous phase (post-manufacture addition), with α-chymotrypsin and aprotinin being the most perturbed, while lipase was relatively unaffected. The effect of zeta potential on immobilisation showed that enzymes adsorbed better closer to their pI, indicating that charge minimisation was necessary for immobilisation. Finally, the effect of increasing enzyme concentration in the aqueous phase resulted in an increase in adsorption for all enzymes due to cooperativity between protein molecules, with saturation occurring faster at higher adsorption rates.

  8. Blood hibernation: a novel strategy to inhibit systemic inflammation and coagulation induced by cardiopulmonary bypass

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jing; WU Xiao-dong; LIN Ke; Raphael C. Lui; AN Qi; TAO Kai-yu; DU Lei; LIU Jin

    2010-01-01

    Background Inflammation and coagulation are two intimately cross-linked defense mechanisms of most, if not all organisms to injuries. During cardiopulmonary bypass (CPB), these two process-is are activated and interact with each other through several common pathways, which may result in subsequent organ dysfunction. In the present study, we hypothesized that the addition of nitric oxide, prostaglandin E1 (PGE1), and aprotinin to the systemic circulation, hereby referred to as blood hibernation, would attenuate the inflammation and coagulation induced by CPB. Methods Thirty adult mongrel dogs were equally divided into five groups, anesthetized and placed on hypothermic CPB (32 C). Each group received respectively the following treatments: (1) inhalation of 40 ppm nitric oxide; (2) intravenous infusion of 20 ng·kg-1·min-1 of PGE1; (3) 80 000 kallikrein inhibitor units (KIU)/kg of aprotinin; (4) the combination of all three agents (blood hibernation group); and (5) no treatment (control group) during CPB. Activation of leukocyte, platelet, endothelial cell, and formation of thrombin were assessed after CPB.Results As compared with the other four groups, leukocyte counts were higher, while plasma elastase, interleukin-8, CD11b mRNA expression, myeloperoxidase activities and lung tissue leukocyte counts were lower in the blood hibernation group (P<0.05 versus other four groups after CPB). Plasma prothrombin fragment (PTF)1+2, and platelet activation factors were lower, while platelet counts were higher in the blood hibernation group (P<0.05 versus other four groups at 6 and 12 hours after CPB). Electron microscopy showed endothelial pseudopods protrusion, with cell adherence in all four groups except the blood hibernation group where endothelial cells remained intact.Conclusion Blood hibernation, effected by the addition of nitric oxide, PGE1 and aprotinin to the circulating blood during extra-corporeal circulation, was observed to attenuate the inflammation and

  9. BESCT (Biology, Education, Screening, Chemoprevention and Treatment) Program

    Science.gov (United States)

    2010-10-01

    Nonidet P - 40 , 1 m M phenylmethylsulfonyl fl uoride, aprotinin at 5 μ g/mL, and leupeptin at 5 μ g/mL. After incubation on ice for 30 minutes and...m M NaCl, 5 m M EDTA, 0.5% Nonidet P - 40 , 5% glycerol, 1 m M phenylmethylsulfonyl fl uoride, and protease inhibitor mixture [Roche diagnostics...buffer (50 mM HEPES (pH 7.5), 150 mM NaCl, 1.5 mM MgCl2, 1 mM EDTA, 0.2 mM EGTA, 1% Nonidet P - 40 , 10% glycerol, 1 mM dithiothreitol, 1 mM

  10. AANA journal course. Update for nurse anesthetists--part 4--life in the balance: the role of serpins in disease genesis and prevention.

    Science.gov (United States)

    Aker, John; Biddle, Chuck

    2007-10-01

    Complex biological systems are often shaped and maintained by opposing forces. A relevant biological example is the delicate balance between proteases and their inhibitors. Serine proteases contain a serine residue in the active site of the molecule that is essential to the activity of the enzyme. Protease inhibitors limit the activity of proteases in the body. As examples, aprotinin (Trasylol), a serine protease inhibitor, and aminocaproic acid (Amicar), a lysine protease inhibitor, are used to decrease the rate of fibrinolysis and have recently been the subject of considerable controversy in the literature regarding safety and efficacy. This AANA journal course reviews 2 common examples of protease inhibitor disorders, angioedema and a form of emphysema, that are of particular anesthetic relevance.

  11. Activation of pro-urokinase and plasminogen on human sarcoma cells

    DEFF Research Database (Denmark)

    Stephens, R W; Pöllänen, J; Tapiovaara, H

    1989-01-01

    Human HT-1080 fibrosarcoma cells produce urokinase-type plasminogen activator (u-PA) and type 1 plasminogen activator inhibitor (PAI-1). We found that after incubation of monolayer cultures with purified native human plasminogen in serum-containing medium, bound plasmin activity could be eluted...... from the cells with tranexamic acid, an analogue of lysine. The bound plasmin was the result of plasminogen activation on the cell surface; plasmin activity was not taken up onto cells after deliberate addition of plasmin to the serum-containing medium. The cell surface plasmin formation was inhibited...... to inhibition by endogenous PAI-1 and by added PAI-2, while the cell-bound plasmin was inaccessible to serum inhibitors, but accessible to added aprotinin and an anticatalytic monoclonal antibody. A model for cell surface plasminogen activation is proposed in which plasminogen binding to cells from serum medium...

  12. Optimization of Cyclic Plasmin Inhibitors: From Benzamidines to Benzylamines.

    Science.gov (United States)

    Hinkes, Stefan; Wuttke, André; Saupe, Sebastian M; Ivanova, Teodora; Wagner, Sebastian; Knörlein, Anna; Heine, Andreas; Klebe, Gerhard; Steinmetzer, Torsten

    2016-07-14

    New macrocyclic plasmin inhibitors based on our previously optimized P2-P3 core segment have been developed. In the first series, the P4 residue was modified, whereas the 4-amidinobenzylamide in P1 position was maintained. The originally used P4 benzylsulfonyl residue could be replaced by various sulfonyl- or urethane-like protecting groups. In the second series, the P1 benzamidine was modified and a strong potency and excellent selectivity was retained by incorporation of p-xylenediamine. Several analogues inhibit plasmin in the subnanomolar range, and their potency against related trypsin-like serine proteases including trypsin itself could be further reduced. Selected derivatives have been tested in a plasma fibrinolysis assay and are more effective than the reference inhibitor aprotinin. The crystal structure of one inhibitor was determined in complex with trypsin. The binding mode reveals a sterical clash of the inhibitor's linker segment with the 99-hairpin loop of trypsin, which is absent in plasmin.

  13. Detection of Metastatic Potential in Breast Cancer by RhoC-GTPase and WISP3 Proteins

    Science.gov (United States)

    2007-05-01

    Lex- ington, KY, USA). Western blot analysis Cells were lysed in lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Nonidet P40 , 1 mM Na3VO4, 1 mM...Available online http://breast-cancer-research.com/content/7/6/R1080 R1084a buffer composed of 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Nonidet ... P40 , 1 mM Na3VO4, 1 mM phenylmethylsulphonyl fluoride, 1 µg/ml leupeptin, and 10 µg/ml aprotinin. The IGF- 1R was immunoprecipitated overnight from

  14. Genomic Instability and Breast Cancer

    Science.gov (United States)

    2009-10-01

    buffer (20 mM Tris-HCl pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5% Nonidet P40 ) containing 50 mM β-glycerophosphate, 10 mM NaF, 1 µg ml−1 pepstatin A and 1 µg...buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40) containing 20 mM NaF and 1 g/ml of pepstatin A and aprotinin on ice for 20 min...irradiated (10 Gy) and harvested 4 h later. Cells were lysed in NETN buffer (20 mM Tris-Hcl at pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40

  15. Activation of Pro-uPA is Critical for Initial Escape from the Primary Tumor and Hematogenous Dissemination of Human Carcinoma Cells

    DEFF Research Database (Denmark)

    Bekes, Erin; Deryugina, Elena; Kuprianova, Tatyana;

    2011-01-01

    Urokinase-type plasminogen activator (uPA) and plasmin have long been implicated in cancer progression. However, the precise contributions of the uPA/plasmin system to specific steps involved in cancer cell dissemination have not been fully established. Herein, we have employed a highly...... disseminating variant of the human PC-3 prostate carcinoma cell line, PC-hi/diss, as a prototype of aggressive carcinomas to investigate the mechanisms whereby pro-uPA activation and uPA-generated plasmin functionally contribute to specific stages of metastasis. The PC-hi/diss cells secrete and activate....... To mechanistically examine the uPA/plasmin-mediated aspects of tumor cell dissemination, the anti pro-uPA mAb-112 and the potent serine protease inhibitor, aprotinin, were utilized in parallel in a number of in vivo assays modeling various rate-limiting steps in early metastatic spread. Our findings demonstrate...

  16. Adsorption of Proteins with Tannin Modified Chitosan Magnetic Particles

    Institute of Scientific and Technical Information of China (English)

    CHANG Yue; SU ZhiXing; WANG YunPu

    2001-01-01

    @@ Magnetic polymer particles have been widely used in biochemistry and medicine in recent years [1-4], mainly due to their property of relatively rapid and easy separation. There were many ways for preparation of magnetic particles [5-9]. We know natural polymer having convenient site such as-NH2,-COOH,-OH,-CONH2, etc. for the affinity ligand attachment. The literature reported chitosan as magnetic polymer matrix, dye as affinity ligand to purify bovine serum albumin and lysozyme[10l. Tannin, a natural product having multiple adjacent hydroxy groups, has extremely high affinity to adsorb protein or alkaloid. However, the information about tannin modified magnetic support is still sparse. Therefore, tannin modified chitosan magnetic particle was prepared and the adsorption of trypsin and aprotinin were studied.

  17. Adsorption of Proteins with Tannin Modified Chitosan Magnetic Particles

    Institute of Scientific and Technical Information of China (English)

    CHANG; Yue

    2001-01-01

    Magnetic polymer particles have been widely used in biochemistry and medicine in recent years [1-4], mainly due to their property of relatively rapid and easy separation. There were many ways for preparation of magnetic particles [5-9]. We know natural polymer having convenient site such as-NH2,-COOH,-OH,-CONH2, etc. for the affinity ligand attachment. The literature reported chitosan as magnetic polymer matrix, dye as affinity ligand to purify bovine serum albumin and lysozyme[10l. Tannin, a natural product having multiple adjacent hydroxy groups, has extremely high affinity to adsorb protein or alkaloid. However, the information about tannin modified magnetic support is still sparse. Therefore, tannin modified chitosan magnetic particle was prepared and the adsorption of trypsin and aprotinin were studied.……

  18. Protein-loaded microspheres prepared by sequential adsorption of dextran sulphate and protamine on melamine formaldehyde core.

    Science.gov (United States)

    Balabushevich, Nadezda G; Larionova, Natalia I

    2009-11-01

    Polyelectrolyte multilayer microspheres were fabricated by layer-by-layer self-assembly of a dextran sulphate and a protamine on melamine formaldehyde cores, followed by the partial decomposition of the core. Effects of pH on the encapsulation of proteins and enzymes with different physico-chemical properties (insulin, aprotinin, peroxidase, glucose oxidase (GOD), catalase (Cat)) in the prepared microspheres were then studied. This method of protein encapsulation demonstrated a high loading capacity and efficiency. The protein incorporation and release was regulated by the pH of the solution. Encapsulated enzymes retained a high specific activity depending on the amount of protein incorporated. Bienzyme system GOD/Cat immobilized in the microspheres was suitable for the glucose content assay.

  19. Role of tissue plasminogen activator/plasmin cascade in delayed neuronal death after transient forebrain ischemia.

    Science.gov (United States)

    Takahashi, Hiroshi; Nagai, Nobuo; Urano, Tetsumei

    We studied the possible involvement of the tissue plasminogen activator (t-PA)/plasmin system on both delayed neuronal death in the hippocampus and the associated enhancement of locomotor activity in rats, after transient forebrain ischemia induced by a four-vessel occlusion (FVO). Seven days after FVO, locomotor activity was abnormally increased and, after 10 days, pyramidal cells were degraded in the CA1 region of the hippocampus. FVO increased the t-PA antigen level and its activity in the hippocampus, which peaked at 4 h. Both the enhanced locomotor activity and the degradation of pyramidal cells were significantly suppressed by intracerebroventricular injection of aprotinin, a plasmin inhibitor, at 4 h but not during FVO. These results suggest the importance of the t-PA/plasmin cascade during the early pathological stages of delayed neuronal death in the hippocampus following transient forebrain ischemia.

  20. The nature of interactions between tissue-type plasminogen activator and platelets

    Energy Technology Data Exchange (ETDEWEB)

    Torr, S.R.; Winters, K.J.; Santoro, S.A.; Sobel, B.E. (Washington Univ., St. Louis, MO (USA))

    1990-07-15

    To elucidate interactions responsible for inhibition of aggregation of platelets in platelet-rich plasma (PRP) harvested from whole blood preincubated with t-PA, experiments were performed with PRP and washed platelets under diverse conditions of preincubation. Both ADP and collagen induced aggregation were inhibited in PRP unless aprotinin had been added to the preincubated whole blood concomitantly with t-PA. However, in washed platelets prepared after the same exposure aggregation was intact. When washed platelets were supplemented with fibrinogen degradation products (FDPs) in concentrations simulating those in whole blood preincubated with t-PA, aggregation induced with either ADP or collagen was inhibited. Thus, the inhibition in PRP depended on generation of FDPs by activated plasminogen. The functional integrity of surface glycoprotein (GP) IIb/IIIa receptors in washed platelets was documented by autoradiography after SDS-PAGE of surface labeled GPs and by fibrinogen binding despite preincubation of the whole blood or washed platelets themselves with t-PA and plasminogen as long as exogenous calcium (greater than or equal to 0.1 microM) was present. In contrast, when calcium was absent, the platelet GP IIb/IIIa receptor was rendered susceptible to degradation by plasmin, and aggregation was inhibited by preincubation at 37 degrees C even if aprotinin was present when aggregation was being assayed. These observations reconcile disparate results in the literature from studies in vivo and in vitro by demonstrating that inhibition of aggregation of platelets in PRP and in whole blood reflects indirect effects of plasminogen activation rather than direct effects of t-PA or plasmin on the platelets themselves.

  1. Purification and characterization of tenerplasminin-1, a serine peptidase inhibitor with antiplasmin activity from the coral snake (Micrurus tener tener) venom.

    Science.gov (United States)

    Vivas, Jeilyn; Ibarra, Carlos; Salazar, Ana M; Neves-Ferreira, Ana G C; Sánchez, Elda E; Perales, Jonás; Rodríguez-Acosta, Alexis; Guerrero, Belsy

    2016-01-01

    A plasmin inhibitor, named tenerplasminin-1 (TP1), was isolated from Micrurus tener tener (Mtt) venom. It showed a molecular mass of 6542Da, similarly to Kunitz-type serine peptidase inhibitors. The amidolytic activity of plasmin (0.5nM) on synthetic substrate S-2251 was inhibited by 91% following the incubation with TP1 (1nM). Aprotinin (2nM) used as the positive control of inhibition, reduced the plasmin amidolytic activity by 71%. Plasmin fibrinolytic activity (0.05nM) was inhibited by 67% following incubation with TP1 (0.1nM). The degradation of fibrinogen chains induced by plasmin, trypsin or elastase was inhibited by TP1 at a 1:2, 1:4 and 1:20 enzyme:inhibitor ratio, respectively. On the other hand, the proteolytic activity of crude Mtt venom on fibrinogen chains, previously attributed to metallopeptidases, was not abolished by TP1. The tPA-clot lysis assay showed that TP1 (0.2nM) acts like aprotinin (0.4nM) inducing a delay in lysis time and lysis rate which may be associated with the inhibition of plasmin generated from the endogenous plasminogen activation. TP1 is the first serine protease plasmin-like inhibitor isolated from Mtt snake venom which has been characterized in relation to its mechanism of action, formation of a plasmin:TP1 complex and therapeutic potential as anti-fibrinolytic agent, a biological characteristic of great interest in the field of biomedical research. They could be used to regulate the fibrinolytic system in pathologies such as metastatic cancer, parasitic infections, hemophilia and other hemorrhagic syndromes, in which an intense fibrinolytic activity is observed.

  2. Efficacy and Safety of Antifibrinolytic Agents in Reducing Perioperative Blood Loss and Transfusion Requirements in Scoliosis Surgery: A Systematic Review and Meta-Analysis.

    Directory of Open Access Journals (Sweden)

    Meng Wang

    Full Text Available Routine use of antifibrinolytic agents in spine surgery is still an issue of debate.To gather scientific evidence for the efficacy and safety of antifibrinolytic agents including aprotinin, tranexamic acid (TXA and epsilon aminocaproic acid (EACA, traditionally known as Amicar in reducing perioperative blood loss and transfusion requirements in scoliosis surgery.We conducted a systematic review and meta-analysis for randomized controlled trials (RCTs, retrospective case-control studies, and retrospective cohort studies on the use of antifibrinolytic agents in scoliosis surgery by searching in the MEDLINE and EMBASE databases and the Cochrane Database of Systematic Reviews and Controlled Trials of papers published from January 1980 through July 2014. Safety of the antifibrinolytic agents was evaluated in all included studies, while efficacy was evaluated in RCTs.Eighteen papers with a total of 1,158 patients were eligible for inclusion in this study. Among them, 8 RCTs with 450 patients were included for evaluation of pharmacologic efficacy (1 RCT was excluded because of a lack of standard deviation data. Mean blood loss was reduced in patients with perioperative use of antifibrinolytic agents by 409.25 ml intraoperatively (95% confidence interval [CI], 196.57-621.94 ml, 250.30 ml postoperatively (95% CI, 35.31-465.30, and 601.40 ml overall (95% CI, 306.64-896.16 ml. The mean volume of blood transfusion was reduced by 474.98 ml (95% CI, 195.30-754.67 ml. The transfusion rate was 44.6% (108/242 in the patients with antifibrinolytic agents and 68.3% (142/208 in the patients with placebo. (OR 0.38; 95% CI; 0.25-0.58; P<0.00001, I2 = 9%. All studies were included for evaluation of safety, with a total of 8 adverse events reported overall (4 in the experimental group and 4 in the control group.The systematic review and meta-analysis indicated that aprotinin, TXA, and EACA all significantly reduced perioperative blood loss and transfusion requirements

  3. On-Chip Separation and MALDI-TOF MS Analysis of His-tagged Protein with NiⅡ-NTA Derivatized Ag Nanoparticles/Porous Silicon Chip%NiⅡ-NTA修饰的银纳米粒/多孔硅芯片在线分离组氨酸标记蛋白和MALDI-TOF质谱检测

    Institute of Scientific and Technical Information of China (English)

    颜红; 王冲; 周小会; 肖守军

    2011-01-01

    An affinity chip was developed via self-assembly of 4-aminothiophenol onto silver nanoparticles (AgNPs)/porous silicon (PSi) chip and chemical conversion of amino groups to Ni Ⅱ-nitrilotriacetic acid (Ni Ⅱ-NTA) termini.The NiⅡ-NTA modified chip was applied to separate and preconcentrate histidine-tagged (his-tagged) fusion proteins,thioredoxin-urodilatin and the lysate of small ubiquitin-related modifier (SUMO)-hu-aprotinin,in a buffer solution containing high levels of salts and solubilizing agents.The on-chip system overcomes the interruption problem of cocrystallization between analytes and matrix molecules due to contaminants from direct injection.It also avoids the complicated off-line pre-treatment of samples.This on-chip separation,purification,and MALDI-TOF MS analysis system is possible to detect target molecules from a complex or an original body fluid solution.%本文通过沉积在多孔硅表面的银纳米粒吸附对氨基苯硫酚和氨基的化学转化得到终端为NiⅡ-Nα,Nα-二(羧甲基)-L-赖氨酸水合物-即Ni Ⅱ-NTA体系的芯片.NiⅡ-NTA修饰的芯片被用于从高浓度的盐和助溶剂的缓冲体系中亲和捕获组氨酸标记的融合蛋白:thioredoxin-urodilatin和SUMO-hu-aprotinin,并进行在线的MALDI-TOF质谱检测,克服了MALDI-TOF质谱中直接点样污染物妨碍样品与基质共结晶的问题,避免了繁琐的离线样品预处理.芯片在线分离、纯化和MALDI-TOF质谱分析体系有望在复杂或原始体液的溶液中分析目标分子.

  4. Whole-saliva proteolysis and its impact on salivary diagnostics.

    Science.gov (United States)

    Thomadaki, K; Helmerhorst, E J; Tian, N; Sun, X; Siqueira, W L; Walt, D R; Oppenheim, F G

    2011-11-01

    There is growing interest in the use of human whole saliva for diagnostics and disease monitoring as an alternative to blood samples. In contrast to blood, whole saliva is a non-sterile body fluid. Proper hand-ling and storage are required to preserve the integrity of potential biomarkers. We investigated salivary autoproteolytic degradation using a variety of approaches. We determined inhibition of protease activities by monitoring the endogenous proteome. In addition, the stability of highly protease-susceptible proteins-histatin 5, statherin, and PRP1-was assessed. Experimental variables included (a) protease inhibitors, (b) salivary pH, (c) incubation temperatures, and (d) sample heating. A cocktail containing AEBSF, aprotinin, pancreatic trypsin inhibitor, leupeptin, antipain, and EDTA could not prevent histatin 5, statherin, or PRP1 degradation in whole saliva. Among the other treatments evaluated, short-term storage of freshly collected samples on ice was effective without interfering with the chemistry of the proteome. In conclusion, whole saliva contains a unique mixture of enzymes as evidenced from their resilience to protease inhibition. Analytical evidence on protein stability is needed to ensure the validity of salivary biomarker study outcomes. Analysis of the data presented will provide help and guidance for the use of saliva samples for diagnostic purposes.

  5. Rational tailoring of substrate and inhibitor affinity via ATRP polymer-based protein engineering.

    Science.gov (United States)

    Murata, Hironobu; Cummings, Chad S; Koepsel, Richard R; Russell, Alan J

    2014-07-14

    Atom transfer radical polymerization (ATRP)-based protein engineering of chymotrypsin with a cationic polymer was used to tune the substrate specificity and inhibitor binding. Poly(quaternary ammonium) was grown from the surface of the enzyme using ATRP after covalent attachment of a protein reactive, water-soluble ATRP-initiator. This "grafting from" conjugation approach generated a high density of cationic ammonium ions around the biocatalytic core. Modification increased the surface area of the protein over 40-fold, and the density of modification on the protein surface was approximately one chain per 4 nm(2). After modification, bioactivity was increased at low pH relative to the activity of the native enzyme. In addition, the affinity of the enzyme for a peptide substrate was increased over a wide pH range. The massively cationic chymotrypsin, which included up to 2000 additional positive charges per molecule of enzyme, was also more stable at extremes of temperature and pH. Most interestingly, we were able to rationally control the binding of two oppositely charged polypeptide protease inhibitors, aprotinin and the Bowman-Birk trypsin-chymotrypsin inhibitor from Glycine max, to the cationic derivative of chymotrypsin. This study expands upon our efforts to use polymer-based protein engineering to predictably engineer enzyme properties without the need for molecular biology.

  6. Oral tolerance induction with altered forms of ovalbumin

    Directory of Open Access Journals (Sweden)

    Stransky B.

    1998-01-01

    Full Text Available As a T cell-dependent phenomenon, oral tolerance is not expected to depend necessarily on native configuration of antigens. We investigated the induction of oral tolerance with modified ovalbumin (Ova. Oral administration of heat-denatured (HD-Ova and cyanogen bromide-degraded ovalbumin was less effective than native Ova in inducing oral tolerance in B6D2F1 mice. HD-Ova was effective in suppressing delayed-type hypersensitivity (DTH reactions but did not suppress specific antibody formation. Injection of Ova directly into the stomach, but not into the ileum or cecum, suppressed subsequent immunization to DTH reactions. Gavage with protease inhibitors (aprotinin or ovomucoid before gavage with Ova was ineffective in blocking tolerance induction. Treatment with hydroxyurea to destroy cycling cells 24 h before gavage with Ova blocked oral tolerance induction and also the possibility to passively transfer tolerance to naive recipients with the serum of mice gavaged with Ova 1 h before. The implications of these findings about oral tolerance induction are discussed

  7. Identification of fibrin clot-bound plasma proteins.

    Directory of Open Access Journals (Sweden)

    Simone Talens

    Full Text Available Several proteins are known to bind to a fibrin network and to change clot properties or function. In this study we aimed to get an overview of fibrin clot-bound plasma proteins. A plasma clot was formed by adding thrombin, CaCl(2 and aprotinin to citrated platelet-poor plasma and unbound proteins were washed away with Tris-buffered saline. Non-covalently bound proteins were extracted, separated with 2D gel electrophoresis and visualized with Sypro Ruby. Excised protein spots were analyzed with mass spectrometry. The identity of the proteins was verified by checking the mass of the protein, and, if necessary, by Western blot analysis. Next to established fibrin-binding proteins we identified several novel fibrin clot-bound plasma proteins, including α(2-macroglobulin, carboxypeptidase N, α(1-antitrypsin, haptoglobin, serum amyloid P, and the apolipoproteins A-I, E, J, and A-IV. The latter six proteins are associated with high-density lipoprotein particles. In addition we showed that high-density lipoprotein associated proteins were also present in fibrinogen preparations purified from plasma. Most plasma proteins in a fibrin clot can be classified into three groups according to either blood coagulation, protease inhibition or high-density lipoprotein metabolism. The presence of high-density lipoprotein in clots might point to a role in hemostasis.

  8. TFPI inhibits lectin pathway of complement activation by direct interaction with MASP-2.

    Science.gov (United States)

    Keizer, Mischa P; Pouw, Richard B; Kamp, Angela M; Patiwael, Sanne; Marsman, Gerben; Hart, Margreet H; Zeerleder, Sacha; Kuijpers, Taco W; Wouters, Diana

    2015-02-01

    The lectin pathway (LP) of complement has a protective function against invading pathogens. Recent studies have also shown that the LP plays an important role in ischemia/reperfusion (I/R)-injury. MBL-associated serine protease (MASP)-2 appears to be crucial in this process. The serpin C1-inhibitor is the major inhibitor of MASP-2. In addition, aprotinin, a Kunitz-type inhibitor, was shown to inhibit MASP-2 activity in vitro. In this study we investigated whether the Kunitz-type inhibitor tissue factor pathway inhibitor (TFPI) is also able to inhibit MASP-2. Ex vivo LP was induced and detected by C4-deposition on mannan-coated plates. The MASP-2 activity was measured in a fluid-phase chromogenic assay. rTFPI in the absence or presence of specific monoclonal antibodies was used to investigate which TFPI-domains contribute to MASP-2 inhibition. Here, we identify TFPI as a novel selective inhibitor of MASP-2, without affecting MASP-1 or the classical pathway proteases C1s and C1r. Kunitz-2 domain of TFPI is required for the inhibition of MASP-2. Considering the role of MASP-2 in complement-mediated I/R-injury, the inhibition of this protease by TFPI could be an interesting therapeutic approach to limit the tissue damage in conditions such as cerebral stroke, myocardial infarction or solid organ transplantation.

  9. Fibrinolysis and anticoagulant potential of a metallo protease produced by Bacillus subtilis K42

    Indian Academy of Sciences (India)

    Wesam A Hassanein; Essam Kotb; Nadia M Awny; Yehia A El-Zawahry

    2011-12-01

    In this study, a potent fibrinolytic enzyme-producing bacterium was isolated from soybean flour and identified as Bacillus subtilis K42 and assayed in vitro for its thrombolytic potential. The molecular weight of the purified enzyme was 20.5 kDa and purification increased its specific activity 390-fold with a recovery of 14%. Maximal activity was attained at a temperature of 40°C (stable up to 65°C) and pH of 9.4 (range: 6.5–10.5). The enzyme retained up to 80% of its original activity after pre-incubation for a month at 4°C with organic solvents such as diethyl ether (DE), toluene (TO), acetonitrile (AN), butanol (BU), ethyl acetate (EA), ethanol (ET), acetone (AC), methanol (ME), isopropanol (IP), diisopropyl fluorophosphate (DFP), tosyl-lysyl-chloromethylketose (TLCK), tosyl-phenylalanyl chloromethylketose (TPCK), phenylmethylsulfonylfluoride (PMSF) and soybean trypsin inhibitor (SBTI). Aprotinin had little effect on this activity. The presence of ethylene diaminetetraacetic acid (EDTA), a metal-chelating agent and two metallo protease inhibitors, 2,2′-bipyridine and -phenanthroline, repressed the enzymatic activity significantly. This, however, could be restored by adding Co2+ to the medium. The clotting time of human blood serum in the presence of this enzyme reached a relative PTT of 241.7% with a 3.4-fold increase, suggesting that this enzyme could be an effective antithrombotic agent.

  10. Nuclear imaging in cardiac amyloidosis

    Energy Technology Data Exchange (ETDEWEB)

    Glaudemans, A.W.J.M.; Slart, R.H.J.A.; Veltman, N.C.; Dierckx, R.A.J.O. [University Medical Center Groningen, Department of Nuclear Medicine and Molecular Imaging, Hanzeplein 1, P.O. Box 30001, Groningen (Netherlands); Zeebregts, C.J. [University Medical Center Groningen, Department of Surgery (Division of Vascular Surgery), Groningen (Netherlands); Tio, R.A. [University Medical Center Groningen, Department of Cardiology, Groningen (Netherlands); Hazenberg, B.P.C. [University Medical Center Groningen, Department of Rheumatology and Clinical Immunology, Groningen (Netherlands)

    2009-04-15

    Amyloidosis is a disease characterized by depositions of amyloid in organs and tissues. It can be localized (in just one organ) or systemic. Cardiac amyloidosis is a debilitating disease and can lead to arrhythmias, deterioration of heart function and even sudden death. We reviewed PubMed/Medline, without time constraints, on the different nuclear imaging modalities that are used to visualize myocardial amyloid involvement. Several SPECT tracers have been used for this purpose. The results with these tracers in the evaluation of myocardial amyloidosis and their mechanisms of action are described. Most clinical evidence was found for the use of {sup 123}I-MIBG. Myocardial defects in MIBG activity seem to correlate well with impaired cardiac sympathetic nerve endings due to amyloid deposits. {sup 123}I-MIBG is an attractive option for objective evaluation of cardiac sympathetic level and may play an important role in the indirect measurement of the effect of amyloid myocardial infiltration. Other, less sensitive, options are {sup 99m}Tc-aprotinin for imaging amyloid deposits and perhaps {sup 99m}Tc-labelled phosphate derivatives, especially in the differential diagnosis of the aetiology of cardiac amyloidosis. PET tracers, despite the advantage of absolute quantification and higher resolution, are not yet well evaluated for the study of cardiac amyloidosis. Because of these advantages, there is still the need for further research in this field. (orig.)

  11. Whole-saliva Proteolysis and Its Impact on Salivary Diagnostics

    Science.gov (United States)

    Thomadaki, K.; Helmerhorst, E.J.; Tian, N.; Sun, X.; Siqueira, W.L.; Walt, D.R.; Oppenheim, F.G.

    2011-01-01

    There is growing interest in the use of human whole saliva for diagnostics and disease monitoring as an alternative to blood samples. In contrast to blood, whole saliva is a non-sterile body fluid. Proper hand-ling and storage are required to preserve the integrity of potential biomarkers. We investigated salivary autoproteolytic degradation using a variety of approaches. We determined inhibition of protease activities by monitoring the endogenous proteome. In addition, the stability of highly protease-susceptible proteins—histatin 5, statherin, and PRP1—was assessed. Experimental variables included (a) protease inhibitors, (b) salivary pH, (c) incubation temperatures, and (d) sample heating. A cocktail containing AEBSF, aprotinin, pancreatic trypsin inhibitor, leupeptin, antipain, and EDTA could not prevent histatin 5, statherin, or PRP1 degradation in whole saliva. Among the other treatments evaluated, short-term storage of freshly collected samples on ice was effective without interfering with the chemistry of the proteome. In conclusion, whole saliva contains a unique mixture of enzymes as evidenced from their resilience to protease inhibition. Analytical evidence on protein stability is needed to ensure the validity of salivary biomarker study outcomes. Analysis of the data presented will provide help and guidance for the use of saliva samples for diagnostic purposes. PMID:21917601

  12. Captopril augments acetylcholine-induced bronchial smooth muscle contractions in vitro via kinin-dependent mechanisms.

    Science.gov (United States)

    Agrawal, Naman; Akella, Aparna; Deshpande, Shripad B

    2016-06-01

    Angiotensin converting enzyme (ACE) inhibitors therapy is aassociated with bothersome dry cough as an adverse effect. The mechanisms underlying this adverse effect are not clear. Therefore, influence of captopril (an ACE inhibitor) on acetylcholine (ACh)-induced bronchial smooth muscle contractions was investigated. Further, the mechanisms underlying the captopril-induced changes were also explored. In vitro contractions of rat bronchial smooth muscle to cumulative concentrations of ACh were recorded before and after exposure to captopril. Further, the involvement of kinin and inositol triphosphate (IP₃) pathways for captopril-induced alterations were explored. ACh produced concentration-dependent (5-500 µM) increase in bronchial smooth muscle contractions. Pre-treatment with captopril augmented the ACh-induced contractions at each concentration significantly. Pre-treatment with aprotinin (kinin synthesis inhibitor) or heparin (inositol triphosphate, IP₃-inhibitor), blocked the captopril-induced augmentation of bronchial smooth muscle contractions evoked by ACh. Further, captopril-induced augmentation was absent in calcium-free medium. These results suggest that captopril sensitizes bronchial smooth muscles to ACh-induced contractions. This sensitization may be responsible for dry cough associated with captopril therapy.

  13. Purification and characterization of a serine protease (CPM-2) with fibrinolytic activity from the dung beetles.

    Science.gov (United States)

    Ahn, Mi Young; Hahn, Bum-Soo; Ryu, Kang Sun; Hwang, Jae Sam; Kim, Yeong Shik

    2005-07-01

    Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gln Asp Phe Val Glu Glu Ile Leu. CPM-2 was inactivated by Cu2+ and Zn2+ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and alpha1-antitrypsin. However, EDTA, EGTA, cysteine, beta-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved Aalpha- and Bbeta-chains of fibrinogen and fibrin, and gamma-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

  14. Protease activities of Acanthamoeba polyphaga and Acanthamoeba castellanii.

    Science.gov (United States)

    Serrano-Luna, José de Jesús; Cervantes-Sandoval, Isaac; Calderón, Jesús; Navarro-García, Fernando; Tsutsumi, Victor; Shibayama, Mineko

    2006-01-01

    Acanthamoeba spp. are free-living amoebae that cause amoebic granulomatous encephalitis, skin lesions, and ocular amoebic keratitis in humans. Several authors have suggested that proteases could play a role in the pathogenesis of these diseases. In the present work, we performed a partial biochemical characterization of proteases in crude extracts of Acanthamoeba spp. and in conditioned medium using 7.5% SDS-PAGE copolymerized with 0.1% m/v gelatin as substrate. We distinguished a total of 17 bands with proteolytic activity distributed in two species of Acanthamoeba. The bands ranged from 30 to 188 kDa in A. castellanii and from 34 to 144 kDa in A. polyphaga. Additionally, we showed that the pattern of protease activity differed in the two species of Acanthamoeba when pH was altered. By using protease inhibitors, we found that the proteolytic activities belonged mostly to the serine protease family and secondly to cysteine proteases and that the proteolytic activities from A. castellanii were higher than those in A. polyphaga. Furthermore, aprotinin was found to inhibit crude extract protease activity on Madin-Darby canine kidney (MDCK) monolayers. These data suggest that protease patterns could be more complex than previously reported.

  15. Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri

    Directory of Open Access Journals (Sweden)

    Mariana Ferreira Silva

    2015-11-01

    Full Text Available Hymenoptera venoms constitute an interesting source of natural toxins that may lead to the development of novel therapeutic agents. The present study investigated the enzymatic and biological characteristics of the crude venom of the ant Odontomachus bauri. Its crude venom presents several protein bands, with higher staining for six proteins with gelatinolytic activity (17, 20, 26, 29, 43 and 48 kDa. The crude venom showed high proteolytic activity on azocasein at optimal pH 8.0 and 37 °C. In the presence of protease inhibitors as aprotinin, leupeptin and EDTA, the azocaseinolytic activity was reduced by 45%, 29% and 9%, respectively, suggesting that the enzymes present in the crude venom belong to the three classes of proteases, with the serine proteases in greater intensity. The crude venom degraded the fibrinogen α-chain faster than the β-chain, while the fibrinogen γ-chain remained unchanged. In biological assays, O. bauri venom showed hemolytic and coagulant activity in vitro, and defibrinating activity in vivo. In addition, the venom showed antimicrobial activity against Staphylococcus aureus and Escherichia coli as well as antiparasitic activity on Toxoplasma gondii infection in vitro. In that sense, this study sheds perspectives for pharmacological applications of O. bauri venom enzymes.

  16. Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri.

    Science.gov (United States)

    Silva, Mariana Ferreira; Mota, Caroline Martins; Miranda, Vanessa dos Santos; Cunha, Amanda de Oliveira; Silva, Maraísa Cristina; Naves, Karinne Spirandelli Carvalho; de Oliveira, Fábio; Silva, Deise Aparecida de Oliveira; Mineo, Tiago Wilson Patriarca; Santiago, Fernanda Maria

    2015-11-30

    Hymenoptera venoms constitute an interesting source of natural toxins that may lead to the development of novel therapeutic agents. The present study investigated the enzymatic and biological characteristics of the crude venom of the ant Odontomachus bauri. Its crude venom presents several protein bands, with higher staining for six proteins with gelatinolytic activity (17, 20, 26, 29, 43 and 48 kDa). The crude venom showed high proteolytic activity on azocasein at optimal pH 8.0 and 37 °C. In the presence of protease inhibitors as aprotinin, leupeptin and EDTA, the azocaseinolytic activity was reduced by 45%, 29% and 9%, respectively, suggesting that the enzymes present in the crude venom belong to the three classes of proteases, with the serine proteases in greater intensity. The crude venom degraded the fibrinogen α-chain faster than the β-chain, while the fibrinogen γ-chain remained unchanged. In biological assays, O. bauri venom showed hemolytic and coagulant activity in vitro, and defibrinating activity in vivo. In addition, the venom showed antimicrobial activity against Staphylococcus aureus and Escherichia coli as well as antiparasitic activity on Toxoplasma gondii infection in vitro. In that sense, this study sheds perspectives for pharmacological applications of O. bauri venom enzymes.

  17. Effect of poly-L-arginine on intestinal absorption of hydrophilic macromolecules in rats.

    Science.gov (United States)

    Yamaki, Tsutomu; Uchida, Masaki; Kuwahara, Yusuke; Shimazaki, Yohei; Ohtake, Kazuo; Kimura, Mitsutoshi; Uchida, Hiroyuki; Kobayashi, Jun; Ogihara, Masahiko; Morimoto, Yasunori; Natsume, Hideshi

    2013-01-01

    We have already reported that poly-L-arginine (PLA) remarkably enhanced the in vivo nasal absorption of hydrophilic macromolecules without producing any significant epithelial damage in rats. In the present study, we examined whether PLA could enhance the absorption of a model hydrophilic macromolecule, fluorescein isothiocyanate-dextran (FD-4), across the intestinal mucosa, as well as the nasal mucosa, by an in situ closed-loop method using the rat intestine. PLA was found to enhance the intestinal absorption of FD-4 in a concentration-dependent manner within the concentrations investigated in this study, but segment-specific differences were found to be associated with this effect (ileum>jejunum>duodenum≧colon). The factors responsible for the segment-specific differences were also investigated by intestinal absorption studies using aprotinin, a trypsin inhibitor, and an analysis of the expression of occludin, a tight junction protein. In the small intestine, the differences in the effect of PLA on the absorption of FD-4 may be related to the enzymatic degradation of PLA. In the colon, the reduced effect of PLA on the absorption of FD-4 may be related to the smaller surface area for absorption and the higher expression of occludin compared with other segments.

  18. Effect of pH and temperature on stability and kinetics of novel extracellular serine alkaline protease (70 kDa).

    Science.gov (United States)

    Bhunia, Biswanath; Basak, Bikram; Mandal, Tamal; Bhattacharya, Pinaki; Dey, Apurba

    2013-03-01

    A novel extracellular serine protease (70 kDa by SDS-PAGE) was purified and characterized. This enzyme retained more than 93% of its initial activity after preincubation for 30 min at 37 °C in the presence of 25% (v/v) tested organic solvents and showed feather degradation activity. The purified enzyme was deactivated at various combinations of pH and temperature to examine the interactive effect of them on enzyme activity. The deactivation process was modeled as first-order kinetics and the deactivation rate constant (k(d)) was found to be minimum at pH 9 and 37 °C. The kinetic analysis of enzyme over a range of pH values indicated two pK values at 6.21 and at 10.92. The lower pK value was likely due to the catalytic histidine in the free enzyme and higher pK value likely reflected deprotonation of the proline moiety of the substrate but ionization of the active site serine is another possibility. Inhibition kinetic showed that enzyme is serine protease because enzyme was competitively inhibited by antipain and aprotinin as these compounds are known to be competitive inhibitors of serine protease. The organic solvent, thermal and pH tolerances of enzyme suggested that it may have potential for use as a biocatalyst in industry.

  19. A Culture-Based Method for Determining the Production of Secreted Protease Inhibitors

    Science.gov (United States)

    Quintero, David; Bermudes, David

    2014-01-01

    We have developed a culture-based method for determining the production of secreted protease inhibitors. The assay utilizes standard proteolysis detection plates to support microbial growth followed by infiltrating the plate with a protease and subsequently detecting the remaining protein by trichloroacetic acid (TCA) precipitation, or by bromocreosol green (BCG) or Ponseau S (PS) staining. The presence of a protease inhibitor can be observed in the form of a protected zone of protein around the protease inhibitor-producing strain. Using the protease inhibitors α-2-macroglobulin, aprotinin, leupeptin, and bestatin and the primary and secondary forms of Photorhabdus luminescens in combination with the protease trypsin, we were able to demonstrate that the assay is specific for the cognate inhibitor of the protease and for bacteria secreting protease inhibitors. In addition, when casein-containing plates were used, the size of the diffusion zone was inversely correlated with the molecular weight of the inhibitor allowing a relative estimation of the protease inhibitor molecular weight. This assay is useful for detecting the presence of microbial secreted protease inhibitors and may reveal their production by microorganisms that were not previously recognized to produce them. PMID:24632514

  20. Purification and characterization of a novel fibrinolytic enzyme from culture supernatant of Pleurotus ostreatus.

    Science.gov (United States)

    Liu, Xiao-Lan; Zheng, Xi-Qun; Qian, Peng-Zhi; Kopparapu, Narasimha-Kumar; Deng, Yong-Ping; Nonaka, Masanori; Harada, Naoki

    2014-02-28

    A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDSPAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the α and β chains of fibrinogen followed by the γ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at 45°C and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

  1. Production and analysis of a biosimilar erythropoietin in Egypt

    Directory of Open Access Journals (Sweden)

    Ebied WM

    2014-05-01

    Full Text Available Wael M Ebied,1 Hytham M Ahmed,2 Fawzy A Elbarbry31SEDICO Pharmaceuticals, Merck & Co External Partner, 6th of October City, Cairo, 2Pharmaceutical Analysis Department, Faculty of Pharmacy, Damanhour University, Damanhour, Egypt; 3Pharmaceutical Sciences, School of Pharmacy, Pacific University Oregon, Hillsboro, OR, USAAbstract: Although management of chronic diseases has been a major challenge for health care systems in developed and developing countries, biopharmaceuticals have been successful in treating many life-threatening conditions. However, the high cost of these agents restricts their availability to countries where patients and/or health care systems are able to afford them. Licensing these biopharmaceuticals as biosimilars after expiration of their patents might increase access to such medicines at an affordable price in developing countries. South Egypt Drug Industries Company (SEDICO is an Egyptian pharmaceutical company that has had the opportunity to manufacture some of these drugs. SEDICO biotechnology products, such as insulin, erythropoietin, streptokinase, angiokinase, follicle-stimulating hormone, aprotinin, filgrastim, and somatropin, have been available on the Egyptian market for more than 6 years. For this paper, erythropoietin, which has been investigated over a number of years, was chosen as a representative example of SEDICO biotechnology products. Our findings confirm that SEDICO erythropoietin can compete with the originator epoetins on the Egyptian market with high quality and at a lower cost.Keywords: biosimilars, developing countries, insulin, human growth hormone, erythropoietin, epoetin, Egypt

  2. An operational research approach to identify cardiac surgery patients at risk of severe post-operative bleeding.

    Science.gov (United States)

    Reddy, Brian; Pagel, Christina; Vuylsteke, Alain; Gerrard, Caroline; Nashef, Sam; Utley, Martin

    2011-09-01

    Severe post-operative bleeding can lead to adverse outcomes for cardiac surgery patients and is a relatively common complication of cardiac surgery. One of the most effective drugs to prevent such bleeding, aprotinin, has been withdrawn from the market due to concerns over its safety. Alternative prophylactic drugs which can be given to patients to prevent bleeding can result in significant side effects and are expensive. For this reason it is difficult to make a clinical or economic case for administering these drugs to all cardiac surgery patients, and the prevailing view is that their use should be targeted at patients considered to be at relatively high risk of post-operative bleeding. However, there is currently no objective method for identifying such patients. Over the past 7 years, a team of clinicians and researchers at Papworth Hospital has collected data concerning post-operative blood loss for each cardiac surgery patient, totalling 11,592 consecutive records. They approached a team of operational researchers (MU, ACP, BR) with extensive experience of developing clinical risk models with the aim of devising a risk stratification scheme that could potentially be used to identify a cohort of higher risk patients. Such patients could be treated with the available prophylactic drugs or recruited to studies to evaluate new interventions. This paper is intended to describe the Operational Research process adopted in the development of this scheme. A concise description of the scheme and its clinical interpretation is published elsewhere.

  3. Characterizing natural hydrogel for reconstruction of three-dimensional lymphoid stromal network to model T-cell interactions.

    Science.gov (United States)

    Kim, Jiwon; Wu, Biming; Niedzielski, Steven M; Hill, Matthew T; Coleman, Rhima M; Ono, Akira; Shikanov, Ariella

    2015-08-01

    Hydrogels have been used in regenerative medicine because they provide a three-dimensional environment similar to soft tissues, allow diffusion of nutrients, present critical biological signals, and degrade via endogenous enzymatic mechanisms. Herein, we developed in vitro system mimicking cell-cell and cell-matrix interactions in secondary lymphoid organs (SLOs). Existing in vitro culture systems cannot accurately represent the complex interactions happening between T-cells and stromal cells in immune response. To model T-cell interaction in SLOs in vitro, we encapsulated stromal cells in fibrin, collagen, or fibrin-collagen hydrogels and studied how different mechanical and biological properties affect stromal network formation. Overall, fibrin supplemented with aprotinin was superior to collagen and fibrin-collagen in terms of network formation and promotion of T-cell penetration. After 8 days of culture, stromal networks formed through branching and joining with other adjacent cell populations. T-cells added to the newly formed stromal networks migrated and attached to stromal cells, similar to the T-cell zones of the lymph nodes in vivo. Our results suggest that the constructed three-dimensional lymphoid stromal network can mimic the in vivo environment and allow the modeling of T-cell interaction in SLOs.

  4. [Epidemiology and prevention of anaphylactoid reactions in heart surgery patients].

    Science.gov (United States)

    Trekova, N A; Solovova, L E; Kuznetsov, R V; Asmangulian, E T

    2000-01-01

    A retrospective analysis of the incidence, severity, and causes of anaphylactoid reactions (AR) in 1504 cardiosurgical patients operated on at Research Center of Surgery in 1995-1999 showed that AR occurred in 109 (7.4%) patients: 60% during aortocoronary bypass operations, 27.2% during correction of acquired heart diseases, and 12.8% during correction of congenital heart diseases. Skin symptoms predominated in the structure of AR (59.7%); cardiovascular episodes ranked second (38.5%), and the incidence of pulmonary reactions was lowest (1.8%). The causes of AR during anesthesia and surgery were platelet-rich plasma and fresh-frozen plasma (35.3%), antibiotics (12.1%), protamine (12.1%), myorelaxants (9.9%), colloid plasma substitutes (8.8%), dioxidine (3.3%), heparin (2.2%), aprotinin (1.1%), diazepam (2.2%), and other agents (7.6%). A history of AR and repeated interventions are risk factors of AR. The protocol of AR prevention in cardiosurgical patients includes determination of risk factors, selection of the least hazardous agents, strict adherence to the rate of infusion of histamine-releasing drugs, minimum utilization of donor blood components (platelets and plasma), use of H1 and H2 blockers, corticosteroids (celestone) during premedication and operation. Such treatment helped decrease the severity and incidence of AR in cardiosurgical patients to 4.7%.

  5. Action of Antiproteases on the Inflammatory Response in Acute Pancreatitis

    Directory of Open Access Journals (Sweden)

    Chun-Chia Chen

    2007-07-01

    Full Text Available The spectrum of acute pancreatitis ranges from mild edematous disease to a severe necrotizing process which is usually accompanied by local or systemic complications and even mortality. Early deaths (within the first week due to severe acute pancreatitis are generally caused by massive inflammatory responses which result in multiple organ failure. Although the exact mechanisms which trigger the inflammatory and necrotizing processes are not completely understood, it is generally accepted that autodigestion and activated leukocytes play important roles in the pathogenesis of acute pancreatitis. Proinflammatory cytokines are associated with systemic inflammatory response syndrome and multiple organ failure syndrome in acute pancreatitis. A compensatory anti-inflammatory response occurs in parallel with systemic inflammatory response syndrome. Trypsin secreted by the pancreatic acinar cells activates proteaseactivated receptor-2 which can result in the production of cytokines. Protease inhibitors such as aprotinin, gabexate mesilate, nafamostat mesilate, ulinastatin, etc. can inhibit the various enzymes and inflammatory response in experimental and clinical studies. Thus, protease inhibitors have been considered as a potential treatment to inhibit the pancreatic inflammation in acute pancreatitis. The beneficial effects of antiproteases on experimental severe acute pancreatitis may be, in part, due to the modulation of inflammatory cytokine responses. The effect of protease inhibitors on the inflammatory response in human acute pancreatitis deserves further study.

  6. Minimizing infarct size. Annual scientific report, 1 Jul 1975--15 Apr 1976

    Energy Technology Data Exchange (ETDEWEB)

    Braunwald, E.

    1976-04-15

    Several goals were achieved during this period of 9 months, both in the experimental laboratory and in patients with acute myocardial infarction. (1) A study of the effects of aprotinin administration on myocardial ischemic injury, subsequent necrosis and collateral blood flow following acute coronary artery occlusion was carried out to completion. (2) A study of the effect of cobra venom factor on myocardial necrosis was completed and the factors responsible for its action were examined. (3) A comparison was made of the effects of nitroglycerin and nitroprusside on ischemic injury and regional myocardial blood flow in patients with acute myocardial infarction and in dogs with coronary occlusions. (4) A method of direct measurement of infarct size in the rat was developed. It consists of occlusion of the main left coronary artery and the histologic quantification of the infarct at 48 hours and 3 weeks later by serial histologic sections or alternatively by measuring total left ventricular myocardial creative phosphokinase activity. (5) New electrocardiographic methods have been developed in order to evaluate atraumatically the extent of myocardial infarction in patients. (6) Intravenous injection of (113)mIn-ENTMP and (99m)TcENTMP in dogs following coronary artery occlusion permitted a sequential double labeling of the damaged myocardium. (7) Since hyaluronidase is a very effective drug in reducing myocardial damage both in the experimental animal and in patients with acute myocardial infarction, a study was carried out to ascertain its effects on collateral flow.

  7. Degradation of extracellular matrix proteins (fibronectin, vitronectin and laminin) by serine-proteinases isolated from Lonomia achelous caterpillar hemolymph.

    Science.gov (United States)

    Lucena, Sara; Guerrero, Belsy; Salazar, Ana M; Gil, Amparo; Arocha-Piñango, Carmen L

    2006-09-01

    Lonomia achelous is a caterpillar distributed in southern Venezuela and in northern Brazil that causes an acute hemorrhagic syndrome in people who have contact with its bristles. The effect of the crude hemolymph and its chromatographic fractions (FDII, Lonomin V and Lonomin V-2) on extracellular matrix proteins was studied. The chromatographic fractions show activities similar to plasmin and urokinase. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, both lonomins appear as a protein band of 25 kDa under reduced conditions. By exclusion chromatography, the molecular weights of Lonomin V and Lonomin V-2 were 26.5 and 24.5 kDa, respectively. Fibronectin, laminin and vitronectin were degraded by all venom components. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, under reduced conditions, shows that lonomins degrade fibronectin in four main fragments of 116, 60, 50 and 30 kDa. Molecular exclusion chromatography in native conditions shows that the molecular masses of these fragments are > or = 300, 62 and 27 kDa. The proteolytic effect of lonomins was abolished by benzamidine/HCl, iodoacetic acid and aprotinin. The extracellular matrix protein degradation together with the fibrino(geno)lytic activity of hemolymph and its fractions could explain, in part, the hemorrhagic syndrome, and the wound dehiscence in persons who have had contact with the L. achelous caterpillar.

  8. Involvement of α2-antiplasmin in dendritic growth of hippocampal neurons.

    Science.gov (United States)

    Kawashita, Eri; Kanno, Yosuke; Asayama, Haruka; Okada, Kiyotaka; Ueshima, Shigeru; Matsuo, Osamu; Matsuno, Hiroyuki

    2013-07-01

    The α2-Antiplasmin (α2AP) protein is known as a principal physiological inhibitor of plasmin, but we previously demonstrated that it acts as a regulatory factor for cellular functions independent of plasmin. α2AP is highly expressed in the hippocampus, suggesting a potential role for α2AP in hippocampal neuronal functions. However, the role for α2AP was unclear. This study is the first to investigate the involvement of α2AP in the dendritic growth of hippocampal neurons. The expression of microtubule-associated protein 2, which contributes to neurite initiation and neuronal growth, was lower in the neurons from α2AP⁻/⁻ mice than in the neurons from α2AP⁺/⁺ mice. Exogenous treatment with α2AP enhanced the microtubule-associated protein 2 expression, dendritic growth and filopodia formation in the neurons. This study also elucidated the mechanism underlying the α2AP-induced dendritic growth. Aprotinin, another plasmin inhibitor, had little effect on the dendritic growth of neurons, and α2AP induced its expression in the neurons from plaminogen⁻/⁻ mice. The activation of p38 MAPK was involved in the α2AP-induced dendritic growth. Therefore, our findings suggest that α2AP induces dendritic growth in hippocampal neurons through p38 MAPK activation, independent of plasmin, providing new insights into the role of α2AP in the CNS.

  9. Endosymbiotic and host proteases in the digestive tract of the invasive snail Pomacea canaliculata: diversity, origin and characterization.

    Directory of Open Access Journals (Sweden)

    Martín S Godoy

    Full Text Available Digestive proteases of the digestive tract of the apple snail Pomacea canaliculata were studied. Luminal protease activity was found in the crop, the style sac and the coiled gut and was significantly higher in the coiled gut. Several protease bands and their apparent molecular weights were identified in both tissue extracts and luminal contents by gel zymography: (1 a 125 kDa protease in salivary gland extracts and in the crop content; (2 a 30 kDa protease throughout all studied luminal contents and in extracts of the midgut gland and of the endosymbionts isolated from this gland; (3 two proteases of 145 and 198 kDa in the coiled gut content. All these proteases were inhibited by aprotinin, a serine-protease inhibitor, and showed maximum activity between 30°C and 35°C and pH between 8.5 and 9.5. Tissue L-alanine-N-aminopeptidase activity was determined in the wall of the crop, the style sac and the coiled gut and was significantly higher in the coiled gut. Our findings show that protein digestion in P. canaliculata is carried out through a battery of diverse proteases originated from the salivary glands and the endosymbionts lodged in the midgut gland and by proteases of uncertain origin that occur in the coiled gut lumen.

  10. 蛋白酶抑制剂对霞水母毒素溶血活性的影响%Effect of the protease inhibitors on the hemolytic activity of jellyfish nematocyst venom

    Institute of Scientific and Technical Information of China (English)

    李荣锋; 于华华; 冯金华; 李鹏程

    2011-01-01

    目的 研究蛋白酶抑制剂对霞水母毒素溶血活性的影响.方法 首先制备霞水母刺丝囊细胞,再利用Mini-Beadbeater组织研磨器破碎细胞提取毒紊,然后向获得的霞水母毒素中加入不同种类及剂量的蛋白酶抑制剂.如金属酶抑制剂EDTA、酸性蛋白酶抑制剂Pepstantin A、丝氨酸蛋白酶抑制剂PMSF和Aprotinin.巯基蛋白酶抑制剂Leupeptin,并且测定蛋白酶抑制剂对霞水母毒素溶血活性的影响.结果 EDTA和Pep-stantin A能够明显抑制蛋白酶的活性从而增强霞水母毒素的溶血活性.1mmo1·L-1EDTA和4μg·m1-1Pepstantin A使其溶血率分别从40%上升到93%和5%上升到78%;而PMSF、Aprotinin和Leupcptin却对霞水母毒素溶血活性的影响较小.结论 金属蛋白酶抑制剂EDTA和酸性蛋白酶抑制刺Pepstantin A能有效地保护霞水母毒素的溶血活性,为深入研究霞水母毒素提供帮助.%Objective To investigate the effect of protease inhibitors on the hemolytic activity of the nematocyst venom of jellyfish Cyanea nozakii Kishinouye. Methods The nematocyst of the jellyfish was prepared and Mini-Beadbeater was used to extract the venom from the nematocyst. Then the effect of protease inhibitors was investigated including EDTA,pepstantin A, PMSF,aprotinin, and leupeptin on the hemolytic activity of the nematocyst venom of jellyfish Cyanea nozakii Kishinouye. Results EDTA and pepstantin A could largely inhibit the activity of the protease in the venom so that the hemolytic activity of the venom could be protected, especially 1 mmol · L-1 EDTA and 4 μg · mL-1 pepstantin A made the hemolytic ratio increase from 40% to 93% and from 5% to 78%, respectively. Conclusion protease inhibitors EDTA and pepstantin A can protect the hemolytic activity of the jellyfish nematocyst venom,which is useful to do further research on the venom.

  11. Cardiac-surgery associated acute kidney injury requiring renal replacement therapy. A Spanish retrospective case-cohort study

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    Garcia-Fernandez Nuria

    2009-09-01

    Full Text Available Abstract Background Acute kidney injury is among the most serious complications after cardiac surgery and is associated with an impaired outcome. Multiple factors may concur in the development of this disease. Moreover, severe renal failure requiring renal replacement therapy (RRT presents a high mortality rate. Consequently, we studied a Spanish cohort of patients to assess the risk factors for RRT in cardiac surgery-associated acute kidney injury (CSA-AKI. Methods A retrospective case-cohort study in 24 Spanish hospitals. All cases of RRT after cardiac surgery in 2007 were matched in a crude ratio of 1:4 consecutive patients based on age, sex, treated in the same year, at the same hospital and by the same group of surgeons. Results We analyzed the data from 864 patients enrolled in 2007. In multivariate analysis, severe acute kidney injury requiring postoperative RRT was significantly associated with the following variables: lower glomerular filtration rates, less basal haemoglobin, lower left ventricular ejection fraction, diabetes, prior diuretic treatment, urgent surgery, longer aortic cross clamp times, intraoperative administration of aprotinin, and increased number of packed red blood cells (PRBC transfused. When we conducted a propensity analysis using best-matched of 137 available pairs of patients, prior diuretic treatment, longer aortic cross clamp times and number of PRBC transfused were significantly associated with CSA-AKI. Patients requiring RRT needed longer hospital stays, and suffered higher mortality rates. Conclusion Cardiac-surgery associated acute kidney injury requiring RRT is associated with worse outcomes. For this reason, modifiable risk factors should be optimised and higher risk patients for acute kidney injury should be identified before undertaking cardiac surgery.

  12. Correlation of Glypican-4 Level with Basal Active Glucagon-Like Peptide 1 Level in Patients with Type 2 Diabetes Mellitus

    Science.gov (United States)

    Koh, Gwanpyo; Cho, Suk Ju; Yoo, So-Yeon; Chin, Sang Ouk

    2016-01-01

    Background Previous studies have reported that glypican-4 (GPC4) regulates insulin signaling by interacting with insulin receptor and through adipocyte differentiation. However, GPC4 has not been studied with regard to its effects on clinical factors in patients with type 2 diabetes mellitus (T2DM). We aimed to identify factors associated with GPC4 level in T2DM. Methods Between January 2010 and December 2013, we selected 152 subjects with T2DM and collected serum and plasma into tubes pretreated with aprotinin and dipeptidyl peptidase-4 inhibitor to preserve active gastric inhibitory polypeptide (GIP) and glucagon-like peptide 1 (GLP-1). GPC4, active GLP-1, active GIP, and other factors were measured in these plasma samples. We performed a linear regression analysis to identify factors associated with GPC4 level. Results The subjects had a mean age of 58.1 years, were mildly obese (mean body mass index [BMI], 26.1 kg/m2), had T2DM of long-duration (mean, 101.3 months), glycated hemoglobin 7.5%, low insulin secretion, and low insulin resistance (mean homeostatic model assessment of insulin resistance [HOMA-IR], 1.2). Their mean GPC4 was 2.0±0.2 ng/mL. In multivariate analysis, GPC4 was independently associated with age (β=0.224, P=0.009), and levels of active GLP-1 (β=0.171, P=0.049) and aspartate aminotransferase (AST; β=–0.176, P=0.043) after being adjusted for other clinical factors. Conclusion GPC4 was independently associated with age, active GLP-1, and AST in T2DM patients, but was not associated with HOMA-IR and BMI, which are well known factors related to GPC4. Further study is needed to identify the mechanisms of the association between GPC4 and basal active GLP-1 levels. PMID:27704740

  13. HIF-2alpha-dependent PAI-1 induction contributes to angiogenesis in hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Geis, Theresa, E-mail: geis@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Döring, Claudia, E-mail: C.Doering@em.uni-frankfurt.de [Dr. Senckenberg Institute of Pathology, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Popp, Rüdiger, E-mail: popp@vrc.uni-frankfurt.de [Institute for Vascular Signalling, Centre for Molecular Medicine, Faculty of Medicine Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60596 Frankfurt am Main (Germany); Grossmann, Nina, E-mail: grossmann@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Fleming, Ingrid, E-mail: fleming@vrc.uni-frankfurt.de [Institute for Vascular Signalling, Centre for Molecular Medicine, Faculty of Medicine Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60596 Frankfurt am Main (Germany); Hansmann, Martin-Leo, E-mail: m.l.hansmann@em.uni-frankfurt.de [Dr. Senckenberg Institute of Pathology, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Dehne, Nathalie, E-mail: dehne@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Brüne, Bernhard, E-mail: b.bruene@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany)

    2015-02-01

    Hypoxia promotes progression of hepatocellular carcinoma (HCC), not only affecting tumor cell proliferation and invasion, but also angiogenesis and thus, increasing the risk of metastasis. Hypoxia inducible factors (HIF)-1α and -2α cause adaptation of tumors to hypoxia, still with uncertainties towards the angiogenic switch. We created a stable knockdown of HIF-1α and HIF-2α in HepG2 cells and generated cocultures of HepG2 spheroids with embryonic bodies as an in vitro tumor model mimicking the cancer microenvironment. The naturally occuring oxygen and nutrient gradients within the cocultures allow us to question the role of distinct HIF isoforms in regulating HCC angiogenesis. In cocultures with a HIF-2α knockdown, angiogenesis was attenuated, while the knockdown of HIF-1α was without effect. Microarray analysis identified plasminogen activator inhibitor 1 (PAI-1) as a HIF-2α target gene in HepG2 cells. The knockdown of PAI-1 in HepG2 cells also lowered angiogenesis. Blocking plasmin, the downstream target of PAI-1, with aprotinin in HIF-2α knockdown (k/d) cells proved a cause–effect relation and restored angiogenesis, with no effect on control cocultures. Suggestively, HIF-2α increases PAI-1 to lower concentrations of active plasmin, thereby supporting angiogenesis. We conclude that the HIF-2α target gene PAI-1 favors the angiogenic switch in HCC. - Highlights: • HepG2 were cocultured with stem cells to mimic a cancer microenvironment in vitro. • A knockdown of HIF-2α reduces angiogenesis. • PAI-1 was identified as a HIF-2α target gene in HCC by microarray analysis. • HIF-2α induces the angiogenic switch via inhibition of plasmin.

  14. Biochemical characterization of a novel fibrinolytic enzyme from Cordyceps militaris.

    Science.gov (United States)

    Liu, Xiaolan; Kopparapu, Narasimha-Kumar; Li, Yao; Deng, Yongping; Zheng, Xiqun

    2017-01-01

    A fibrinolytic enzyme was produced by the medicinal mushroom, Cordyceps militaris using submerged fermentation. The enzyme was purified from culture supernatant by hydrophobic interaction, ion exchange and gel filtration chromatographies. It was purified by 36 fold, with a specific activity of 1,467.4U/mg protein and the final yield was 5.8%. The molecular weight of the enzyme as determined by SDS-PAGE and gel filtration was 28kDa and 24.5kDa, respectively, and its isoelectric point (pI) was 9.0±0.2. It was found to be a glycoprotein with carbohydrate content of 1.67% (w/v). The enzyme was optimally active at 37°C and pH 7.2. The enzyme activity was strongly inhibited by soybean trypsin inhibitor (SBTI) and aprotinin which indicated it to be a serine protease, while other inhibitors like N-α-tosyl-l-phenylalanine chloromethyl ketone (TPCK), phenyl methane sulfonyl fluoride (PMSF), pepstatin and metal chelator EDTA did not inhibit its activity. Amino acid sequences of the purified enzyme were determined partially by Q-TOF2 and they were IEDFPYQVDLR; ANCGGTVISEK; YVLTAGHCAEGYTGLNIR; TNYASVTPITADMICAGFPEGK; KDSCSGDSGGPLVTGGK; VVGIVSFGTGCAR; ANKPGVYSSVASAEIR. Sequences of the seven peptides completely matched with those of a trypsin-like serine protease from Cordyceps militaris CM01 (accession no. EGX95217.1). The purified enzyme degraded α chains of fibrinogen first and then β and γ chains and also activated plasminogen into plasmin. It can act as an anticoagulant and prevent clot formation by degrading fibrinogen. Based on these studies, the purified enzyme has great potential to be developed as a natural agent for prevention and treatment of thrombolytic diseases.

  15. Inhibition of tryptase and chymase induced nucleated cell infiltration by proteinase inhibitors

    Institute of Scientific and Technical Information of China (English)

    Shao-heng HE; Han-qiu CHEN; Jian ZHENG

    2004-01-01

    AIM: To investigate the ability of proteinase inhibitors to modulate nucleated cell infiltration into the peritoneum of mice induced by tryptase and chymase. METHODS: Human lung tryptase and skin chymase were purified by a similar procedure involving high salt extraction, heparin agarose affinity chromatography followed by S-200 Sephacryl gel filtration chromatography. The actions of proteinase inhibitors on tryptase and chymase induced nucleated cell accumulation were examined with a mouse peritoneum model. RESULTS: A selective chymase inhibitor Z-Ile-GluPro-Phe-CO2Me (ZIGPPF) was able to inhibit approximately 90% neutrophil, 73% eosinophil, 87% lymphocyte and 60% macrophage accumulation induced by chymase at 16 h following injection. Soy bean trypsin inhibitor (SBTI), chymostatin, and α1-antitrypsin showed slightly less potency than ZIGPPF in inhibition of the actions of chymase. While all tryptase inhibitors tested were able to inhibit neutrophil, eosinophil, and macrophage accumulation provoked by tryptase at 16 h following injection, only leupeptin, APC366, and aprotinin were capable of inhibiting tryptase induced lymphocyte accumulation. The inhibitiors of tryptase tested were also able to inhibit tryptase induced neutrophil and eosinophil accumulation at 6 h following injection. When being injected alone, all inhibitors of chymase and tryptase at the concentrations tested by themselves had no significant effect on the accumulation of nucleated cells in the peritoneum of mice at both 6 h and 16 h. CONCLUSION: Proteinase inhibitors significantly inhibited tryptase and chymase-induced nucleated cell accumulation in vivo, and therefore they are likely to be developed as a novel class of anti-inflammatory drugs.

  16. A novel signaling pathway of tissue kallikrein in promoting keratinocyte migration: Activation of proteinase-activated receptor 1 and epidermal growth factor receptor

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Lin; Chao, Lee [Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425-2211 (United States); Chao, Julie, E-mail: chaoj@musc.edu [Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425-2211 (United States)

    2010-02-01

    Biological functions of tissue kallikrein (TK, KLK1) are mainly mediated by kinin generation and subsequent kinin B2 receptor activation. In this study, we investigated the potential role of TK and its signaling pathways in cultured human keratinocyte migration and in a rat skin wound healing model. Herein, we show that TK promoted cell migration and proliferation in a concentration- and time-dependent manner. Inactive TK or kinin had no significant effect on cell migration. Interestingly, cell migration induced by active TK was not blocked by icatibant or L-NAME, indicating an event independent of kinin B2 receptor and nitric oxide formation. TK's stimulatory effect on cell migration was inhibited by small interfering RNA for proteinase-activated receptor 1 (PAR{sub 1}), and by PAR{sub 1} inhibitor. TK-induced migration was associated with increased phosphorylation of epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK), which was blocked by inhibition of protein kinase C (PKC), Src, EGFR and ERK. TK-induced cell migration and EGFR phosphorylation were blocked by metalloproteinase (MMP) inhibitor, heparin, and antibodies against EGFR external domain, heparin-binding EGF-like growth factor (HB-EGF) and amphiregulin (AR). Local application of TK promoted skin wound healing in rats, whereas icatibant and EGFR inhibitor blocked TK's effect. Skin wound healing was further delayed by aprotinin and neutralizing TK antibody. This study demonstrates a novel role of TK in skin wound healing and uncovers new signaling pathways mediated by TK in promoting keratinocyte migration through activation of the PAR{sub 1}-PKC-Src-MMP pathway and HB-EGF/AR shedding-dependent EGFR transactivation.

  17. Vacuolar targeting of r-proteins in sugarcane leads to higher levels of purifiable commercially equivalent recombinant proteins in cane juice.

    Science.gov (United States)

    Palaniswamy, Harunipriya; Syamaladevi, Divya P; Mohan, Chakravarthi; Philip, Anna; Petchiyappan, Anushya; Narayanan, Subramonian

    2016-02-01

    Sugarcane is an ideal candidate for biofarming applications because of its large biomass, rapid growth rate, efficient carbon fixation pathway and a well-developed storage tissue system. Vacuoles occupy a large proportion of the storage parenchyma cells in the sugarcane stem, and the stored products can be harvested as juice by crushing the cane. Hence, for the production of any high-value protein, it could be targeted to the lytic vacuoles so as to extract and purify the protein of interest from the juice. There is no consensus vacuolar-targeting sequence so far to target any heterologous proteins to sugarcane vacuole. Hence, in this study, we identified an N-terminal 78-bp-long putative vacuolar-targeting sequence from the N-terminal domain of unknown function (DUF) in Triticum aestivum 6-SFT (sucrose: fructan 6-fructosyl transferase). In this study, we have generated sugarcane transgenics with gene coding for the green fluorescent protein (GFP) fused with the vacuolar-targeting determinants at the N-terminal driven by a strong constitutive promoter (Port ubi882) and demonstrated the targeting of GFP to the vacuoles. In addition, we have also generated transgenics with His-tagged β-glucuronidase (GUS) and aprotinin targeted to the lytic vacuole, and these two proteins were isolated and purified from the transgenic sugarcane and compared with commercially available protein samples. Our studies have demonstrated that the novel vacuolar-targeting determinant could localize recombinant proteins (r-proteins) to the vacuole in high concentrations and such targeted r-proteins can be purified from the juice with a few simple steps.

  18. Stability of glucagon-like peptide 1 and glucagon in human plasma

    Science.gov (United States)

    Wewer Albrechtsen, Nicolai J; Bak, Monika J; Hartmann, Bolette; Christensen, Louise Wulff; Kuhre, Rune E; Deacon, Carolyn F; Holst, Jens J

    2015-01-01

    To investigate the stability of glucagon-like peptide 1 (GLP-1) and glucagon in plasma under short- and long-term storage conditions. Pooled human plasma (n=20), to which a dipeptidyl peptidase 4 (DPP4) inhibitor and aprotinin were added, was spiked with synthetic GLP-1 (intact, 7–36NH2 as well as the primary metabolite, GLP-1 9–36NH2) or glucagon. Peptide recoveries were measured in samples kept for 1 and 3 h at room temperature or on ice, treated with various enzyme inhibitors, after up to three thawing–refreezing cycles, and after storage at −20 and −80 °C for up to 1 year. Recoveries were unaffected by freezing cycles or if plasma was stored on ice for up to 3 h, but were impaired when samples stood at RT for more than 1 h. Recovery of intact GLP-1 increased by addition of a DPP4 inhibitor (no ice), but was not further improved by neutral endopeptidase 24.11 inhibitor or an inhibitor cocktail. GLP-1, but not glucagon, was stable for at least 1 year. Surprisingly, the recovery of glucagon was reduced by almost 50% by freezing compared with immediate analysis, regardless of storage time. Plasma handling procedures can significantly influence results of subsequent hormone analysis. Our data support addition of DPP4 inhibitor for GLP-1 measurement as well as cooling on ice of both GLP-1 and glucagon. Freeze–thaw cycles did not significantly affect stability of GLP-1 or glucagon. Long-term storage may affect glucagon levels regardless of storage temperature and results should be interpreted with caution. PMID:25596009

  19. New options in the management of tendinopathy

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    Nicola Maffulli

    2010-03-01

    Full Text Available Nicola Maffulli1, Umile Giuseppe Longo2, Mattia Loppini2, Filippo Spiezia2, Vincenzo Denaro21Centre for Sports and Exercise Medicine, Queen Mary University of London, Barts and The London School of Medicine and Dentistry, Mile End Hospital, London, England; 2Department of Orthopedic and Trauma Surgery, Campus Biomedico University, Rome, ItalyAbstract: Tendon injuries can be acute or chronic, and caused by intrinsic or extrinsic factors, either alone or in combination. Tendinopathies are a common cause of disability in occupational medicine and account for a substantial proportion of overuse injuries in sports. Tendinopathy is essentially a failed healing response, with haphazard proliferation of tenocytes, abnormalities in tenocytes, with disruption of collagen fibres and subsequent increase in noncollagenous matrix. The scientific evidence base for managing tendinopathies is limited. What may appear clinically as an “acute tendinopathy” is actually a well advanced failure of a chronic healing response in which there is neither histologic nor biochemical evidence of inflammation. In this review we report the new options for the management of tendinopathy, including eccentric exercises, extracorporeal shockwave therapy, injections (intratendinous injections of corticosteroids, aprotinin, polidocanol platelet-rich plasma, autologous blood injection, high-volume injections and surgery. Open surgery aims to excise fibrotic adhesions, remove areas of failed healing and make multiple longitudinal incisions in the tendon to detect intratendinous lesions, and to restore vascularity and possibly stimulate the remaining viable cells to initiate cell matrix response and healing. New surgical techniques aim to disrupt the abnormal neoinnervation to interfere with the pain sensation caused by tendinopathy. These procedures are intrinsically different from the classical ones in present use, because they do not attempt to address directly the pathologic

  20. Partial biochemical and biological characterization of purified chicken growth hormone (cGH). Isolation of cGH charge variants and evidence that cGH is phosphorylated.

    Science.gov (United States)

    Arámburo, C; Carranza, M; Sanchez, R; Perera, G

    1989-11-01

    Chicken growth hormone (cGH) was purified from frozen pituitary glands obtained from recently sacrificed broilers. Glands were homogenized in a protease inhibitor solution (0.5 mM PMSF, 50 KIU/ml aprotinin, pH 7.2); extract was taken to pH 9.0 with calcium hydroxide and the supernatant was differentially precipitated with 20% (fraction A) and 50% (fraction B) ammonium sulfate. cGH (fraction B-DE-1) was obtained in pure form from fraction B after DEAE-cellulose chromatography at pH 8.6, with a yield of 2.9 mg/g tissue. Three charge variants of cGH (Rf = 0.23, 0.30, and 0.35) could be isolated by electroelution after semipreparative nondenaturing polyacrylamide gel electrophoresis of fraction B-DE-1. These charge variants showed the same apparent molecular weight (26,300 Da) by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions. Isoelectric focusing of fraction B-DE-1 revealed two major components (pI = 7.2 and 7.4) and four minor bands (pI = 6.2, 6.7, 7.1, and 7.5). It was found that fraction B-DE-1 contained a significant amount of esterified phosphate (1 nmol PO4/3.5 nmol protein) similar to that reported previously for ovine GH. The functional integrity of the cGH obtained here was characterized by two heterologous and one homologous bioassays. High activity was shown by fraction B-DE-1 in the tibia assay (1.76 UI/mg) and in the liver ornithine decarboxylase assay (sixfold over control), both made in hypophysectomized rats; and it also stimulated lipolysis (138 and 215% at 10 and 100 ng/ml, respectively) on chicken abdominal adipose tissue explants.

  1. Purification and biochemical characterization of a novel fibrinolytic enzyme, PSLTro01, from a medicinal animal Porcellio scaber Latreille.

    Science.gov (United States)

    Tian, Zhou; Li, Bo; Guo, Liwei; Wu, Mianhua; Fu, Tingming; Cheng, Haibo; Zhu, Huaxu

    2015-09-01

    A novel protease, named PSLTro01, with fibrinolytic and anticoagulant activity was isolated from Porcellio scaber Latreille and was purified by a combination of hollow fibre membrane molecular weight cut-off (MWCO), ammonium sulfate fractionation, gel filtration and ion-exchange chromatography. PSLTro01 is a single-chain protein with a molecular mass of 38,497 Da as estimated by non-reduced SDS-PAGE and MALDI-TOF MS spectrometry, and its N-terminal 15 amino acid sequence was determined as DINGGGATLPQPLYQ. PSLTro01 is stable in the range of 20-40 °C and pH 6.0-10.0, with a maximum fibrinolytic activity at 40 °C and pH 7.0. The PSLTro01-induced fibrinolytic activity was not influenced by K(+) or Na(+) but was slightly increased by Mg(2+) and completely inhibited by aprotinin and pepstatin A. Fibrin plate assays revealed that PSLTro01 could not directly degrade fibrin but was a plasminogen activator. PSLTro01 exhibited high specificity for the substrate S-2251 for plasmin, followed by S-2238 for thrombin and S-2444 for urokinase. Moreover, the fibrinogenolysis pattern of PSLTro01 was Aα-chains>Bβ-chains>γ-chain. Tail-thrombus of the enzyme treated group was significantly shorter than the physiological saline treated group and the thrombus decrement was correlated with the enzyme dose. PSLTro01 prolongs both thrombin time (TT) and activated partial thromboplastin time (APTT). These results indicate that PSLTro01 may have potential applications in the prevention and treatment of thrombosis.

  2. Strategies to prevent intraoperative lung injury during cardiopulmonary bypass

    Directory of Open Access Journals (Sweden)

    Siminelakis Stavros N

    2010-01-01

    Full Text Available Abstract During open heart surgery the influence of a series of factors such as cardiopulmonary bypass (CPB, hypothermia, operation and anaesthesia, as well as medication and transfusion can cause a diffuse trauma in the lungs. This injury leads mostly to a postoperative interstitial pulmonary oedema and abnormal gas exchange. Substantial improvements in all of the above mentioned factors may lead to a better lung function postoperatively. By avoiding CPB, reducing its time, or by minimizing the extracorporeal surface area with the use of miniaturized circuits of CPB, beneficial effects on lung function are reported. In addition, replacement of circuit surface with biocompatible surfaces like heparin-coated, and material-independent sources of blood activation, a better postoperative lung function is observed. Meticulous myocardial protection by using hypothermia and cardioplegia methods during ischemia and reperfusion remain one of the cornerstones of postoperative lung function. The partial restoration of pulmonary artery perfusion during CPB possibly contributes to prevent pulmonary ischemia and lung dysfunction. Using medication such as corticosteroids and aprotinin, which protect the lungs during CPB, and leukocyte depletion filters for operations expected to exceed 90 minutes in CPB-time appear to be protective against the toxic impact of CPB in the lungs. The newer methods of ultrafiltration used to scavenge pro-inflammatory factors seem to be protective for the lung function. In a similar way, reducing the use of cardiotomy suction device, as well as the contact-time between free blood and pericardium, it is expected that the postoperative lung function will be improved.

  3. Endogenously generated plasmin at the vascular wall injury site amplifies lysine binding site-dependent plasminogen accumulation in microthrombi.

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    Tomasz Brzoska

    Full Text Available The fibrinolytic system plays a pivotal role in the regulation of hemostasis; however, it remains unclear how and when the system is triggered to induce thrombolysis. Using intra-vital confocal fluorescence microscopy, we investigated the process of plasminogen binding to laser-induced platelet-rich microthrombi generated in the mesenteric vein of transgenic mice expressing green fluorescent protein (GFP. The accumulation of GFP-expressing platelets as well as exogenously infused Alexa Fluor 568-labeled Glu-plasminogen (Glu-plg on the injured vessel wall was assessed by measuring the increase in the corresponding fluorescence intensities. Glu-plg accumulated in a time-dependent manner in the center of the microthrombus, where phosphatidylserine is exposed on platelet surfaces and fibrin formation takes place. The rates of binding of Glu-plg in the presence of ε-aminocaproic acid and carboxypeptidase B, as well as the rates of binding of mini-plasminogen lacking kringle domains 1-4 and lysine binding sites, were significantly lower than that of Glu-plg alone, suggesting that the binding was dependent on lysine binding sites. Furthermore, aprotinin significantly suppressed the accumulation of Glu-plg, suggesting that endogenously generated plasmin activity is a prerequisite for the accumulation. In spite of the endogenous generation of plasmin and accumulation of Glu-plg in the center of microthrombi, the microthrombi did not change in size during the 2-hour observation period. When human tissue plasminogen activator was administered intravenously, Glu-plg further accumulated and the microthrombi were lysed. Glu-plg appeared to accumulate in the center of microthrombi in the early phase of microthrombus formation, and plasmin activity and lysine binding sites were required for this accumulation.

  4. Interaction of human malignant melanoma (ST-ML-12) tumor spheroids with endothelial cell monolayers. Damage to endothelium by oxygen-derived free radicals.

    Science.gov (United States)

    Offner, F. A.; Wirtz, H. C.; Schiefer, J.; Bigalke, I.; Klosterhalfen, B.; Bittinger, F.; Mittermayer, C.; Kirkpatrick, C. J.

    1992-01-01

    Clinical and experimental observations suggest that tumor-induced endothelial cell injury may be one of several initial events in the establishment of tumor metastases. To test this hypothesis, the authors have analyzed the interaction of malignant melanoma (ST-ML-12) multicenter tumor spheroids with endothelial cell monolayers in a three-dimensional coculture system. After 1.5 hours of interaction, the authors observed a toxic effect on endothelial cells in the perispheroid region. The latter was demonstrated by testing membrane integrity with the fluorescent probes acridine orange/ethidium bromide and resulted in sensitivity to shear stress of the damaged cells. The endothelium then underwent a regenerative cycle to replace the denuded halo. Addition of the oxygen radical-scavenging enzyme superoxide dismutase to the culture medium prevented this endothelial cell damage in a dose-dependent manner for up to 12 hours. By contrast, catalase, deferoxamine mesylate, allopurinol, and the proteinase inhibitors soybean trypsin inhibitor and aprotinin were not protective under the same conditions. The endothelial damage was dependent on the attachment of the spheroids. Medium conditioned by ST-ML-12-spheroids proved to be ineffective. A similar, but less prominent, deleterious effect was seen when human peritoneal mesothelial cells were used in place of the human umbilical vein endothelial cells. Spheroids of the uroepithelial cell line HU-609 were used as control. No toxicity was observed in these cocultures. Melanin biosynthesis is associated with the production of oxygen-derived free radicals. The results suggest a possible implication of these free radicals in metastasis formation of malignant melanoma. Images Figure 1 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 PMID:1519667

  5. Increased nociceptin/orphanin FQ plasma levels in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Ferenc Szalay; Mónika B Hantos; Andrea Horvath; Peter L. Lakatos; Aniko Folhoffer; Kinga Dunkel; Dalma Hegedus; Kornélia Tekes

    2004-01-01

    AIM: The heptadecapeptide nociceptin alias orphanin FQ is the endogenous agonist of opioid receptor-like1 receptor.It is involved in modulation of pain and cognition. High blood level was reported in patients with acute and chronic pain,and in Wilson disease. An accidental observation led us to investigate nociceptin in hepatocellular carcinoma.METHODS: Plasma nociceptin level was measured by radioimmunoassay, aprotinin was used as protease inhibitor.Hepatocellular carcinoma was diagnosed by laboratory,ultrasound, other imaging, and confirmed by fine needle biopsy. Results were compared to healthy controls and patients with other chronic liver diseases.RESULTS: Although nociceptin levels were elevated in patients with Wilson disease (14.0±2.7 pg/mL, n=26),primary biliary cirrhosis (12.1±3.2 pg/mL, n=21) and liver cirrhosis (12.8±4.0 pg/mL, n=15) compared to the healthy controls (9.2±1.8 pg/mL, n=29, P<0.001 for each), in patients with hepatocellular carcinoma a ten-fold increase was found (105.9±14.4 pg/mL, n=29, P<0.0001). High plasma levels were found in each hepatocellular carcinoma patient including those with normal alpha fetoprotein and those with pain (104.9±14.9 pg/mL, n=12) and without (107.7±14.5pg/mL, n=6).CONCLUSION: A very high nociceptin plasma level seems to be an indicator for hepatocellular carcinoma. Further research is needed to clarify the mechanism and clinical significance of this novel finding.

  6. Concentration-Dependent Dual Role of Thrombin In Protection of Cultured Rat Cortical Neurons

    Science.gov (United States)

    García, Paul S.; Ciavatta, Vincent T.; Fidler, Jonathan A.; Woodbury, Anna; Levy, Jerrold H.; Tyor, William R.

    2015-01-01

    Background Thrombin’s role in the nervous system is not well understood. Under conditions of blood-brain barrier compromise (e.g., neurosurgery or stroke), thrombin can result in neuroapoptosis and the formation of glial scars. Despite this, preconditioning with thrombin has been found to be neuroprotective in models of cerebral ischemia and intracerebral hemorrhage. Methods We investigated the effects of physiologically relevant concentrations of thrombin on cortical neurons using two culture-based assays. We examined thrombin’s effect on neurites by quantitative analysis of fluorescently labeled neurons. To characterize thrombin’s effects on neuron survival, we spectrophotometrically measured changes in enzymatic activity. Using receptor agonists and thrombin inhibitors, we separately examined the role of thrombin and its receptor in neuroprotection. Results We found that low concentrations of thrombin (1 nM) enhances neurite growth and branching, neuron viability, and protects against excitotoxic damage. In contrast, higher concentrations of thrombin (100 nM) are potentially detrimental to neuronal health as evidenced by inhibition of neurite growth. Lower concentrations of thrombin resulted in equivalent neuroprotection as the antifibrinolytic, aprotinin, and the direct thrombin inhibitor, argatroban. Interestingly, exogenous application of the species-specific thrombin inhibitor, antithrombin III, was detrimental to neuronal health; suggesting that some endogenous thrombin is necessary for optimal neuron health in our culture system. Activation of the thrombin receptor, protease-activated receptor - 1 (PAR-1), via micromolar concentrations of the thrombin receptor agonist peptide, TRAP, did not adversely affect neuronal viability. Conclusions An optimal concentration of thrombin exists to enhance neuronal health. Neurotoxic effects of thrombin do not involve activation of PAR receptors and thus separate pharmacologic manipulation of thrombin’s receptor

  7. Effect of naloxone on level of plasma beta-endorphin in neonates with severe asphyxia

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    BACKGROUND: β-endorphin is the most actively endogenous substance of cerebral endorphin. When combined with opiate receptor specially, it manifests a strong morphine-like activity and can decrease sensitivity of central nervous system to carbon dioxide so as to inhibit breath.OBJECTIVE: To observe the changes of content of plasma β -endorphin in neonates with severe asphyxia after naloxone treatment in a large dosage.DESIGN: Randomized controlled observation.SETTINGS: Department of Pediatrics, Shenzhen Shajing People's Hospital; Center of Pediatrics,Guangzhou Zhujiang Hospital.PARTICIPANTS: A total of 97 neonates with severe asphyxia including 57 boys and 40 girls were selected from Neonatal Intensive Care Unit, Department of Pediatrics, Shenzhen Shajing People's Hospital from January 2004 to November 2005. Their gestational age was (38 ±3) weeks, body mass was (3.2± 1.7) kg,and hospitalization duration was (2.8±2.3) hours. All neonates met the diagnostic criteria of with severe asphyxia and all their parents provided the confirmed consent.METHODS: All neonates were treated with inspired oxygen, sedation, stopping terror, decreasing cranial pressure, maintaining a well blood perfusion and normal level of blood glucose (about 5.0 mmol/L). After hospitalization, 0.1 mg/(kg ·d) naloxone hydrochloride (Beijing Sihuan Pharmaceutical Technology Co., Ltd.;certification: H10900021; bullet preparation; 0.4 mg/ampoule) was intravenously dribbled into neonates for 4 - 6 hours, 14 days in total. 2 mL blood was collected from radial artery in neonates at the beginning of hospitalization and at 3 days after naloxone treatment, put in aprotinin-pre-cool tube, mixed evenly, and centrifuged at hypothermia. Plasma was maintained in refrigerator at - 70 ℃. The kit was provided by Neurobiology Department of Shanghai Second Military Medical University of Chinese PLA. Concentration of plasma β -endorphin was measured by using radio-immunity assay.All data were expressed as Mean

  8. 抑肽酶以剂量依赖性方式影响小鼠缺血-再灌注后左心室收缩功能和细胞因子释放

    Institute of Scientific and Technical Information of China (English)

    Matthew D.McEvoy; Rupak Mukherjee; Francis G.Spinale; 黄瑛; Michel J.Sabbagh; Anna Greta Taylor; Juozas A.Zavadzkas; Christine N.Koval; Robert E.Stroud; Rachael L.Ford; Julie E.McLean; Scott T.Reeves

    2012-01-01

    背景 心脏手术期间阶段性的缺血-再灌注(ischemia-reperfusion,I/R)常伴随短暂性左室(left ventricular,LV)功能障碍和炎症反应.本研究探讨了I/R情况下丝氨酸蛋白酶抑制剂抑肽酶(potential dose-dependent effects of aprotinin,APRO)影响LV收缩功能和细胞因子释放的可能剂量依赖性效应.方法 应用微传感器容量分析法测定基础水平、缺血30分钟,再灌注60分钟时的LV收缩功能指数,左心室最大弹性(maximal elastance,Emax).小鼠随机分组为:(a) APRO 20 000激肽释放酶抑制单位(KIU)/kg组(n=11); (b)APRO4×104KIU/kg组(n=10);(c) APRO 8×104KIU/kg组(n=10);(d)溶剂组(生理盐水;n=10).计算的APRO用量反映临床哈默史密斯剂量表用量的半量、1个剂量和2个剂量.采集I/R后的动物血浆测定细胞因子浓度.结果 I/R后溶剂组以及APRO 4×104KIU/kg组和APRO 8×104KIU/kg组动物Emax较基础水平下降超过40%(P<0.05).然而,APRO 2×104KIU/kg组Emax回到接近基础水平.I/R后血浆肿瘤坏死因子(tumor necrosis factor,TNF)升高10倍,但是TNF随着APRO剂量的增加而下降.结论 本研究结果证实,低剂量的APRO对LV收缩功能具有保护作用,而较高剂量APRO可抑制TNF释放.这些独创性的研究结果提示,APRO在I/R情况下具有其不同的独立的作用机制.

  9. Prospective observational study for perioperative volume replacement with 6% HES 130/0,42, 4% gelatin and 6% HES 200/0,5 in cardiac surgery

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    Winterhalter M

    2010-09-01

    Full Text Available Abstract Background The constantly growing amount of different kinds of colloid fluids necessitates comparative investigations with regards to the safety and effectivity in clinical use of these preparations. Hence we compared three colloid fluids in an observational study. The objective was the exploration of the influence of these three colloids on blood coagulation, hemodynamics and renal function of the cardiac surgical patient. Methods We included 90 patients undergoing an elective open-heart surgery with the use of the heart-lung machine and observed them consecutively. Group 1 [gelatin 4% (n = 30], Group 2 [HES 200/0,5 (n = 30] and Group 3 [HES 130/0,42 (n = 30]. We measured the perioperative volume replacement, the administration of blood- and coagulation-products, the application of catecholamines, the renal function, blood gas and the platelet aggregation using multiplate electrode analyzer (Multiplate®, Dynabyte medical, Munich, Germany. Results The gelatin-group needed significantly more norepinephrine than the HES 130/0.42 group. The responsible surgeon considered the blood coagulation in the HES 200/0.5 group most frequently as impaired. Furthermore we saw a significant decrease in platelet function in the HES 200/0.5 group when performing the multiplate®-analysis (ADP-and COL-test. HES 130/0.4 as well as gelatin 4% showed no significant change in platelet function. The gelatin-group and the HES 200/0.5 needed significantly more aprotinine than the HES 130/0.4 group. We saw no significant difference with regards to administration of blood and coagulation products between the three groups. The urinary excretion during the intervention was significantly higher in the HES 200/0.5 group and in the gelatin group than in the HES 130/0.4 group. Conclusions Our results confirm the lower stabilizing effect of gelatin on circulation during fluid resuscitation. The blood coagulation was mostly impaired due to HES 200/0.5 confirmed by the

  10. Angiogenic synergistic effect of basic fibroblast growth factor and vascular endothelial growth factor in an in vitro quantitative microcarrier-based three-dimensional fibrin angiogenesis system

    Institute of Scientific and Technical Information of China (English)

    Xi-Tai Sun; Yi-Tao Ding; Xiao-Gui Yan; Ling-Yun Wu; Qiang Li; Ni Cheng; Yu-Dong Qiu; Min-Yue Zhang

    2004-01-01

    AIM: To develop an in vitro three-dimensional (3-D)angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and to quantify their synergistic activity.METHODS: Microcarriers (MCs) coated with human microvascular endothelial cells (HMVECs) were embedded in fibrin gel and cultured in 24-well plates with assay media. The growth factors bFGF, or VEGF, or both were added to the system. The wells (n = 8/group) were digitally photographed and the average length of capillary-like sprouts (ALS) from each microcarrier was quantitated.RESULTS: In aprotinin-stabilized fibrin matrix, human microvascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina. The angiogenic effects of bFGF or VEGF were dose-dependent in the range from 10 to 40 ng/mL. At d 1, 10 ng/mL of bFGF and VEGF induced angiogenesis with an ALS of 32.13±16.6 μm and 43.75±27.92 μm, respectively, which were significantly higher than that of the control (5.88±4.45 μm, P<0.01),and the differences became more significant as the time increased. In addition, the combination of 10 ng/mL of bFGF and VEGF each induced a more significant effect than the summed effects of bFGF (10 ng/mL) alone and VEGF (10 ng/mL) alone when analyzed using SPSS system for general linear model (GLM) (P= 0.011), and that also exceeded the effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION: A microcarrier-based in vitro threedimensional angiogenesis model can be developed in fibrin.It offers a unique system for quantitative analysis of angiogenesis. Both bFGF and VEGF exert their angiogenic effects on HMVECs synergistically and in a dose-dependent manner.

  11. Effect of antifibrinolytic drugs on transfusion requirement and blood loss during orthotopic liver transplantation: Results from a single center

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    Devi A

    2008-01-01

    Full Text Available Background: During orthotopic liver transplantation (OLT, activation of the fibrinolytic system can contribute significantly to perioperative bleeding. Prophylactic administration of antifibrinolytic agents has been shown to reduce blood loss and the need for allogenic transfusion. Objective: To study the effect of antifibrinolytics on requirement of blood components, blood loss and operative time during OLT in patients with end stage liver disease, reporting to a single centre. Materials and Methods: Consecutive patients who underwent OLT at this centre during the period February 2003-October 2007 were the subjects of this study. Based on the individual anesthesiologist′s preference, patients were assigned to receive either two million units of aprotinin (AP as a bolus followed by 5,00,000 units/hour or 10 mg/kg tranexamic acid (TAas a bolus followed by 10 mg/kg every six to eight hours, administered from the induction till the end of the surgery. Transfusion policy was standardized in all patients. Intraoperative red cell salvage was done wherever possible. The effect of these two antifibrinolytic drugs on transfusion requirement was evaluated as a whole and in a sub group of patients from each treatment group and compared with a concurrent control group that did not receive antifibrinolytic drugs. Results: Fifty patients (40 M / 10 F, 44 adults, 6 pediatric patients underwent OLT in the study period. Fourteen patients were given AP, 25 patients were given TA and 11 patients did not receive any of the agents(control group. The median volume of total blood components transfused in antifibrinolytic group (n=39 was 4540 ml(0-19,200ml, blood loss 5 l(0.7-35l and operative time 9h (4.5-17h and that of control group(n=11 was 5700 ml(0-15,500ml, 10 l(0.6-25 l and 9h (6.4-15.8h respectively. The median volume of blood transfusions, blood loss and operative time was lesser in AP group(n=14 than that of TA group(n=25. Conclusion: There is definite

  12. 高脂饮食叙利亚金黄地鼠胰岛局部血管紧张素Ⅱ的生成途径研究%Pathways of local angiotensin II generation in islets of Langerhans in Syrian golden hamster fed with high-fat diet

    Institute of Scientific and Technical Information of China (English)

    叶文慧; 孙侃; 孙嘉; 张桦; 陈宏; 蔡德鸿

    2011-01-01

    AIM : To investigate the diverse pathways of local angiotensin Ⅱ ( Ang Ⅱ ) generation in islets of Langerhans in Syrian golden hamsters with dyslipidemia.METHODS : The Syrian golden hamsters were fed with high - fat diet to induce dyslipidemia.Purified islet cells from dyslipidemia and normal Syrian hamsters were prepared and divided into control group, captopril group, chymotrypsin endostatin group, aprotinin group, α - antitrypsin group and captopril +chymotrypsin endostatin group by adding respective reagents into the cultured cells after treated with angiotensin Ⅰ.The Ang Ⅱ levels in the supernatants of each group were examined hy ELISA.RESULTS : Compared with the c:ontrol animals, Ang Ⅱ levels decreased in all groups with interventions.Compared with the normal hamsters islet cells, the Ang Ⅱ levels in captopril group, chymotrypsin endostatin group, α - antitrypsin group, aprotinin group and captopril + chymotrypsin endostatin group were decreased by 39.98% , 50.10% ( P< 0.01 ), 23.04%, 20.85% ( P < 0.05 ) and 82.78% ( P <0.01 ), respectively.Compared with high - fat group, the corresponding data were 42.12% , 56.96% ( P < 0.01 ),26.11% . 22.68% ( P < 0.05 ) and 83.59% ( P <0.01 ), respectively.The levels of Ang Ⅱ in chymotrypsin group between normal and high - fat diet hamsters were significantly different ( P < 0.05 ).CONCLUSION : Under the condition of dyslipidemia, the classic angiotensin - converting enzyme - based pathway and chymotrypsin pathway are still the main approaches of producing Ang Ⅱ in male Syrian hamster islet to produce angiotensin.The effect of chymotrypsin endostatin is comparatively stronger in inhibiting the production of local Ang Ⅱ than the effect of angiotensin - converting enzyme inhihitor.%目的:观察不同抑制剂阻断局部肾素血管紧张素系统(RAS)后,对血脂谱异常血症叙利亚金黄地鼠胰岛细胞生成血管紧张素Ⅱ(AngⅡ)的影响,探讨高脂饮

  13. The Impact of Pancreatic Enzyme Supplementation on Postprandial Responses of Glucagon-Like Peptide-2 in Patients with Chronic Pancreatitis and Pancreatic Exocrine Insufficiency

    Directory of Open Access Journals (Sweden)

    Filip K Knop

    2010-09-01

    from our 2007 study [8]. Eight patients with chronic pancreatitis and pancreatic exocrine insufficiency were investigated using two liquid meal tests consumed with and without two capsules of pancreatic enzyme supplementation (Creon® 25,000 U, Solvay Pharma, Herlev, Denmark as previously described [8]. A group of 8 healthy sexage- and body mass index-matched control subjects was examined using the same meal. Arterialized blood was drawn and distributed into EDTA tubes containing aprotinin and a dipeptidyl peptidase 4 (DPP4 inhibitor as previously described [8]. The tubes were centrifuged for 20 minutes at 1,700 g and 4°C, and plasma was stored at -20 °C until analysis. The GLP-2 radioimmunoassay employs antiserum code no. 92160 and standards of human GLP-2 (proglucagon 126-158, a gift from Novo Nordisk A/S, Bagsværd, Denmark and monoiodinated Tyr-12 GLP-1, specific activity greater than 70 MBq/nmol. The antiserum is directed against the N-terminus of GLP-2 and therefore measures only fully processed intact GLP-2 of intestinal origin. Sensitivity for the assay is below 2 pmol/L and the intra-assay coefficient of variation is below 6% [9]. Data are expressed as mean±SEM and were compared by means of analysis of variance (ANOVA. Area under curve (AUC values were calculated using the trapezoidal rule.

  14. Capacidade da matriz extracelular da medula óssea de induzir proliferação de células mielóides in vitro no modelo de desnutrição protéica em camundongos Capacity of the extracellular matrix of the bone marrow to induce proliferation of myeloid cells in vitro in model of protein malnutrition in mice

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    Cidônia de Lourdes Vituri

    2008-09-01

    Full Text Available Este trabalho tem por objetivo verificar se a matriz extracelular (MEC obtida da medula óssea de camundongos com desnutrição protéica energética sustenta a sobrevivência, se induz proliferação de células mielóides, bem como avaliar a capacidade desta MEC de interagir com citocinas hematopoiéticas in vitro. Camundongos machos "Swiss" foram submetidos à desnutrição protéica (4% de caseína até que perdessem 20% do peso inicial e o grupo-controle foi mantido com uma dieta contendo 14% de caseína. A medula óssea foi extraída com tampão PBS suplementado com 1 mg de aprotinina/mL. Os ensaios de proliferação foram realizados com a linhagem mielóide FDC-P1, pelo método colorimétrico de redução do MTT. A MEC obtida tanto do grupo-controle como do desnutrido induziu proliferação celular in vitro. Os ensaios de interação foram realizados com IL-3 e GM-CSF na concentração de 10 ρg e 500 ρg/mL, que demonstraram efeito sinérgico e efeito regulatório, respectivamente. A MEC obtida de animais do grupo desnutrido quando submetida ao ensaio de ligação ao GM-CSF mostrou maior proliferação celular do que a MEC obtida de animais do grupo-controle (pThe aim of this study was to verify the capacity of the extracellular matrix (ECM obtained from bone marrow of malnourished mice to sustain survival and to induce the proliferation of myeloid cells. We also verified the capacity of the tests to interact with in vitro hematopoietic cytokines. Male "Swiss" mice were submitted to protein malnutrition with a diet content of '4% casein until they lost 20% of the original weight, while the group-control was kept with a diet content of 14% of casein. The bone marrow was extracted with 1.0 mg of aprotinin/mL in PBS. The proliferation tests were carried out with myeloid cell line FDCP-1, by the colorimetric method of reduction of the MTT. The obtained ECM from nourished and undernourished mice induced cellular proliferation invitro. Tests

  15. Regulación por proteasas del canal de sodio sensible al amiloride (ENaC Amiloride sensitive sodium channels (ENaC and their regulation by proteases

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    Luciano Galizia

    2011-04-01

    regulate the activity of ENaC channels: 1 the number of channels inserted in the membrane and 2 the open probability of the channels or time that the channel is open. The number of channels is the result of a balance between the synthesis and degradation of ENaC channels. The open probability depends on the proteolysis of specific segments in the α and γ subunits of ENaC by multiple proteases inside of the cell or in the extracellular space. Among the most studied proteases are furin, prostasin, elastase, plasmin and trypsin. There are endogenous substances that block the activity of these proteases such as aprotinin, bikunin and nexin-1 and the expression of both, proteases and their inhibitors are controlled by the rate of Na+ movement, aldosterone and TFG-β levels. In this work we present some examples of this regulation and the potential role that this process may play under normal and pathological conditions such as cystic fibrosis, kidney diseases and hypertension.

  16. 冠心病糖代谢异常患者血浆Ghrelin水平及临床意义%Plasma ghrelin level in patients with coronary heart disease with abnormal glucose metabolism and its clinical significance

    Institute of Scientific and Technical Information of China (English)

    庞军刚; 徐新; 唐良秋; 张社兵; 江志平

    2012-01-01

    目的:探讨冠心病糖代谢异常患者血浆胃饥饿素(Ghrelin)水平及其相关临床意义.方法:将纳入研究对象依据相关检验及检查结果分为正常对照组、冠心病组(冠心病糖代谢正常组和冠心病糖代谢异常组)、单纯糖代谢异常组.收集所有入选对象人院第2天清晨空腹血样,采用ELISA方法同批检测血浆Ghrelin水平.结果:①冠心病组及单纯糖代谢异常组血浆Ghrelin水平均显著低于正常对照组.②冠心病糖代谢异常组血浆Ghrelin水平显著低于冠心病糖代谢正常组及单纯糖代谢异常组.③析因分析结果显示:冠心病与糖代谢异常在对血浆Ghrelin水平影响方面不存在交互作用.然而,糖代谢异常比冠心病对血浆Ghrelin水平的影响更明显.结论:冠心病糖代谢异常患者血浆Ghrelin水平显著下降,且糖代谢异常对Ghrelin的影响更明显.%AIM: To study plasma ghrelin level distribution in patients with coronary heart disease (CHD) with abnormal glucose metabolism and to discuss its clinical significance. METHODS; According to laboratory examination results, subjects were divided into control group, coronary heart disease with normal glucose metabolism group, coronary heart disease with abnormal glucose metabolism group and abnormal glucose metabolism group. Fasting blood samples were collected the morning after admission with EDTA-2K anticoagulation tubes. Blood samples were then transferred to centrifuge tubes containing aprotinin and were centrifuged to extract plasma for cryopreservation. All blood plasma ghrelin levels were tested with ELISA. RESULTS: Compared with those in control group, ghrelin levels were significantly reduced in the group with CHD with normal glucose metabolism, group of CHD with abnormal glucose metabolism and group with abnormal glucose metabolism. Compared with those in the group of CHD with normal glucose metabolism, levels of ghrelin were significantly reduced in patients with

  17. Rol del sistema kallicreína kinina y su interrelación con sistemas vasoactivos durante la preñez Role of the kallikrein kinin system and its interrelationship with vasoactive systems in pregnancy

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    Elisabet Oddo

    2011-10-01

    systems. It was demonstrated in rats that activation of KKS precedes the installation of glomerular hyperfiltration as aprotinin prevents the increase in glomerular filtration. In addition, individual or associated inhibition of specific kallikrein kinin system effectors, prostaglandins (PGs and nitric oxide (NO, confirm the glomerular filtration rate dependence of KKS during pregnancy. It was also found that the renin-angiotensin system (RAS contributes to glomerular hyperfiltration as this is affected by the administration of RAS blockers. The peak of hyperfiltration maximum inhibition was obtained by the blockade of both systems (KKS and RAS. In addition, strategies used to alter the glomerular hyperfiltration and increased sodium reabsorption during pregnancy, showed abnormalities in the development of the fetus and placenta, fewer offspring, more fetus resorptions and intrauterine growth retardation. KKS inhibitors associated with RAS or nitric oxide blockers showed the greatest impact. As a consequence, it was demonstrated that KKS plays a central role in the adaptation phenomenom that accompanies normal pregnancy. The interplay of KKS with several vasoactive systems, seem to arrange a network involved in the hemodynamic adaptations to allow the proper development of pregnancy and the fetus and placenta.

  18. Anestesia para o recém-nascido submetido a cirurgia cardíaca com circulação extracorpórea Anestesia para el recién nacido sometido a cirugía cardiaca con circulación extracorpórea Anesthesia for the newborn submitted to cardiac surgery with cardiopulmonary bypass

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    Sérgio Bernardo Tenório

    2005-02-01

    ítrico o los inhibidores de la fosfodiesterasa. CONCLUSIONES: El anestesista tiene papel preponderante en el ajuste de la homeostasia durante el período peri-operatorio. Conocimientos sobre el tipo de lesión cardiaca, la corrección a ser realizada, la respuesta del organismo a la CEC pueden ser útiles en el manoseo de estos niños.BACKGROUND AND OBJECTIVES: Congenital heart diseases affect 0.8% of liveborn infants and many need neonatal surgical correction. Cardiac surgery with cardiopulmonary bypass (CPB in this age is associated to higher risk of complications related to child's functional immaturity, lack of CPB equipment fully compatible with neonate (NN size and technical difficulties to correct cardiac defects. This article aimed at describing aspects related to anesthetic technique, CPB and their effects on NN. CONTENTS: High fentanyl or sufentanil doses promote adequate anesthesia without interfering with cardiocirculatory stability. Opioids residual respiratory depression is not a problem for these patients because most of them will need immediate postoperative respiratory assistance. CPB may be followed by heart manipulation-induced hypotension and/or bleeding. Inadequate venous and aortic cannula position may lead to severe complications, such as insufficient brain flow or difficult venous drainage. Deep hypothermia and total circulatory arrest are common during CPB. Hypothermia changes blood viscosity, which is treated with hemodilution and has implications on pH correction (alpha-stat versus pH stat. Low cardiac output is common during CPB weaning and adjustments in one or all its components (preload, contractility, afterload and heart rate may be necessary. In addition to classic drugs, such as epinephrine and dopamine, other substances may be needed, such as aprotinin, nitric oxide or phosphodiesterase inhibitors. CONCLUSIONS: Anesthesiologists play a major role in adjusting perioperative homeostasis. Understanding the type of cardiac disease, the

  19. Guidelines, Editors, Pharma And The Biological Paradigm Shift

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    Ajai R. Singh

    2007-01-01

    Full Text Available Private investment in biomedical research has increased over the last few decades. At most places it has been welcomed as the next best thing to technology itself. Much of the intellectual talent from academic institutions is getting absorbed in lucrative positions in industry. Applied research finds willing collaborators in venture capital funded industry, so a symbiotic growth is ensured for both. There are significant costs involved too. As academia interacts with industry, major areas of conflict of interest especially applicable to biomedical research have arisen. They are related to disputes over patents and royalty, hostile encounters between academia and industry, as also between public and private enterprise, legal tangles, research misconduct of various types, antagonistic press and patient-advocate lobbies and a general atmosphere in which commercial interest get precedence over patient welfare.Pharma image stinks because of a number of errors of omission and commission. A recent example is suppression of negative findings about Bayer's Trasylol (Aprotinin and the marketing maneuvers of Eli Lilly's Xigris (rhAPC. Whenever there is a conflict between patient vulnerability and profit motives, pharma often tends to tilt towards the latter. Moreover there are documents that bring to light how companies frequently cross the line between patient welfare and profit seeking behaviour. A voluntary moratorium over pharma spending to pamper drug prescribers is necessary. A code of conduct adopted recently by OPPI in India to limit pharma company expenses over junkets and trinkets is a welcome step.Clinical practice guidelines (CPG are considered important as they guide the diagnostic/therapeutic regimen of a large number of medical professionals and hospitals and provide recommendations on drugs, their dosages and criteria for selection. Along with clinical trials, they are another area of growing influence by the pharmaceutical industry. For

  20. Guidelines, editors, pharma and the biological paradigm shift.

    Science.gov (United States)

    Singh, Ajai R; Singh, Shakuntala A

    2007-01-01

    Private investment in biomedical research has increased over the last few decades. At most places it has been welcomed as the next best thing to technology itself. Much of the intellectual talent from academic institutions is getting absorbed in lucrative positions in industry. Applied research finds willing collaborators in venture capital funded industry, so a symbiotic growth is ensured for both.There are significant costs involved too. As academia interacts with industry, major areas of conflict of interest especially applicable to biomedical research have arisen. They are related to disputes over patents and royalty, hostile encounters between academia and industry, as also between public and private enterprise, legal tangles, research misconduct of various types, antagonistic press and patient-advocate lobbies and a general atmosphere in which commercial interest get precedence over patient welfare.Pharma image stinks because of a number of errors of omission and commission. A recent example is suppression of negative findings about Bayer's Trasylol (Aprotinin) and the marketing maneuvers of Eli Lilly's Xigris (rhAPC). Whenever there is a conflict between patient vulnerability and profit motives, pharma often tends to tilt towards the latter. Moreover there are documents that bring to light how companies frequently cross the line between patient welfare and profit seeking behaviour.A voluntary moratorium over pharma spending to pamper drug prescribers is necessary. A code of conduct adopted recently by OPPI in India to limit pharma company expenses over junkets and trinkets is a welcome step.Clinical practice guidelines (CPG) are considered important as they guide the diagnostic/therapeutic regimen of a large number of medical professionals and hospitals and provide recommendations on drugs, their dosages and criteria for selection. Along with clinical trials, they are another area of growing influence by the pharmaceutical industry. For example, in a

  1. Monograph: Concluding Remarks

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    Ajai R. Singh

    2007-01-01

    Full Text Available Private investment in biomedical research has increased over the last few decades. At most places it has been welcomed as the next best thing to technology itself. There are significant costs involved too. Major areas of conflict of interest especially applicable to biomedical research have arisen, as academia interacts with industry. Pharma image stinks because of a number of errors of omission and commission. A recent example is suppression of negative findings about Bayer's Trasylol (Aprotinin and the marketing maneuvers of Eli Lilly's Xigris (rhAPC. A voluntary moratorium over pharma spending to pamper drug prescribers is necessary.The integrity of industry-sponsored clinical research has come under increasing scrutiny. The basic shizm is between the value system of a patient welfare driven professional and that of a profit driven industry . While one wants to avoid control but wants the dough, the other wants to exercise the control by supplying the dough. Clinical practice guidelines (CPG are considered important as they guide diagnostic/therapeutic regimen of a large number of medical professionals and hospitals and provide recommendations on drugs, dosages and criteria for selection. Besides clinical trials, they are another area of growing influence by the pharmaceutical industry. For example, in a recent survey it was found that about 60% of 192 authors of clinical practice guidelines reported they had financial connections with the companies whose drugs were under consideration. This finding casts serious doubt on the credibility of this important pillar of modern clinical practice. It needs urgent reparative action. One of them is prospective and retrospective disclosure of financial conflict of interest by authors of CPGs.A Conference on Guideline Standardization (COGS was convened in April 2002 'to define a standard for guideline reporting that would promote guideline quality and facilitate implementation'. It includes items for