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Sample records for aprotinin

  1. Aprotinin and major orthopedic surgery

    OpenAIRE

    Samama, Charles Marc

    2004-01-01

    Aprotinin is a potent pharmacological agent that reduces bleeding and limits blood transfusion requirements in current surgical practice. Many studies have been conducted in orthopedic surgery. In several trials performed in total hip replacement (THR) and total knee replacement (TKN) patients, aprotinin only moderately decreased blood-loss-replacement requirements. Conversely, when aprotinin was used in patients at high risk for bleeding (cancer, sepsis, redone surgery), it developed a poten...

  2. Affinity purification of aprotinin from bovine lung.

    Science.gov (United States)

    Xin, Yu; Liu, Lanhua; Chen, Beizhan; Zhang, Ling; Tong, Yanjun

    2015-05-01

    An affinity protocol for the purification of aprotinin from bovine lung was developed. To simulate the structure of sucrose octasulfate, a natural specific probe for aprotinin, the affinity ligand was composed of an acidic head and a hydrophobic stick, and was then linked with Sepharose. The sorbent was then subjected to adsorption analysis with pure aprotinin. The purification process consisted of one step of affinity chromatography and another step of ultrafiltration. Then purified aprotinin was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, trypsin inhibitor activity, gel-filtration, and thin-layer chromatography analysis. As calculated, the theoretical maximum adsorption (Qmax ) of the affinity sorbent was 25,476.0 ± 184.8 kallikrein inactivator unit/g wet gel; the dissociation constant of the complex "immobilized ligand-aprotinin" (Kd ) was 4.6 ± 0.1 kallikrein inactivator unit/mL. After the affinity separation of bovine lung aprotinin, reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis and gel-filtration chromatography revealed that the protein was a single polypeptide, and the purities were ∼ 97 and 100%, respectively; the purified peptide was also confirmed with aprotinin standard by gel-filtration chromatography and thin-layer chromatography. After the whole purification process, protein, and bioactivity recoveries were 2.2 and 92.6%, respectively; and the specific activity was up to 15,907.1 ± 10.2 kallikrein inactivator unit/mg. PMID:25677462

  3. Successful Management of Tendinopathy With Injections of the MMP-inhibitor Aprotinin

    OpenAIRE

    Orchard, John; Massey, Andrew; Brown, Richard; Cardon-Dunbar, Adéline; Hofmann, Jamie

    2008-01-01

    Aprotinin is a broad spectrum proteinase inhibitor (including matrix metalloproteinase [MMP] inhibitor) used for treating patellar and Achilles tendinopathies. One previous randomized control trial demonstrated aprotinin injections superior to both corticosteroid and saline injections in patellar tendinopathy (Level II), whereas results reported for aprotinin treatment in Achilles tendinopathy have been mixed. We performed a case review and followup questionnaire for 430 consecutive patients ...

  4. Aprotinin inhibits the hormone binding of the estrogen receptor from calf uterus.

    Science.gov (United States)

    Nigro, V; Medici, N; Abbondanza, C; Minucci, S; Molinari, A M; Puca, G A

    1989-11-15

    Micromolar concentrations of the proteinase inhibitor, aprotinin, produced a dose-dependent inhibition in the binding capacity of the estrogen receptor from calf uterus. Aprotinin inhibition was greater at 28 degrees C than at 4 degrees C and only occurred when conditions allowed the receptor transformation. When aprotinin was tested in the presence of transformation inhibitors, its effect was no longer seen. The binding capacity of the highly purified estrogen-binding subunit was similarly inhibited. PMID:2480113

  5. 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...

  6. An aprotinin binding site localized in the hormone binding domain of the estrogen receptor from calf uterus.

    Science.gov (United States)

    Nigro, V; Medici, N; Abbondanza, C; Minucci, S; Moncharmont, B; Molinari, A M; Puca, G A

    1990-07-31

    It has been proposed that the estrogen receptor bears proteolytic activity responsible for its own transformation. This activity was inhibited by aprotinin. Incubation of transformed ER with aprotinin modified the proteolytic digestion of the hormone binding subunit by proteinase K. The smallest hormone-binding fragment of the ER, obtained by tryptic digestion, was still able to bind to aprotinin. These results suggest that aprotinin interacts with ER and the hormone-binding domain of ER is endowed with a specific aprotinin-binding site. PMID:1696480

  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. Influence of ultra-low dose Aprotinin on thoracic surgical operations: a prospective randomized trial

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    Sourgiadaki Efrosini

    2008-03-01

    Full Text Available Abstract Background The blood saving effect of aprotinin has been well documented in cardiac surgery. In thoracic surgery, very few recent studies, using rather high doses of aprotinin, have shown a similar result. In a randomized prospective trial, we have tested the influence of aprotinin using an ultra-low dose drug regime. Methods Fifty-nine patients, mean age 58 ± 13.25 years (mean ± SD undergoing general thoracic procedures were randomized into placebo (Group A and treatment group (Group B. The group B (n = 29 received 500.000 IU of aprotinin after induction to anesthesia and a repeat dose immediately after chest closure. A detailed protocol with several laboratory parameters was recorded. Patients were transfused when perioperative Ht was less than 26%. Results The two groups were similar in terms of age, gender, diagnosis, pathology, co-morbidity and operations performed. The mean drainage of the first and second postoperative day in group B was significantly reduced (412.6 ± 199.2 vs. 764.3 ± 213.9 ml, p Conclusion The perioperative ultra-low dose aprotinin administration was associated with a reduction of total blood losses and blood product requirements. We therefore consider the use of aprotinin safe and effective in major thoracic surgery.

  9. Efficacy of aprotinin treatment on bilateral blunt chest trauma created in rabbits

    International Nuclear Information System (INIS)

    Objectives: To investigate the effects of aprotinin, on blood gasses, oxidant-antioxidant status, and lung histopathology in an experimental bilateral blunt chest trauma model. Methods: Conducted at the Experimental Animal Laboratory of Meram Medical School at Selcuk University, Konya, Turkey, the study comprised 21 New Zealand female albino rabbits who were divided into three groups. Trauma was applied on the sham and aprotinin groups, which was administered intravenous Aprotinin 20.000 U/kg. Arterial blood samples were obtained from all rabbits at hours 0, 3, 24, and 96. At hour 96 after trauma, all rabbits were sacrificed using the decapitation method, and then blood and lung tissue samples were obtained. Blood nitric oxide, malondialdehyde and blood gas measurements were made. Histopathological changes in the lung were examined with a light microscope. Results: While no positive effect of aprotinin was observed on nitric oxide malondialdehyde and partial pressure of carbon dioxide values, it was seen to have an increasing effect on partial oxygen pressure level. Aprotinin had a partial effect on lung histopathology. Conclusion: Aprotinin was determined to have a positive effect on PO/sub 2/ levels. We could not find any positive effects especially on alveolar haemorrhage. (author)

  10. The epidermal growth factor precursor in the rat kidney seems to be processed by an aprotinin sensitive proteinase

    DEFF Research Database (Denmark)

    Nexø, Ebba; Poulsen, Steen Seier; Raaberg, Lasse

    1992-01-01

    Epidermal growth factor (EGF) is synthesized as a membrane bound precursor in the rat kidney. The precursor seems to be processed by an aprotinin sensitive proteinase. Intravenous infusion of aprotinin reduces the urinary excretion of EGF by 85% and increases the amount of renal EGF. Kidney...

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. The influence of aprotinin on regional absorption of soluble human insulin.

    OpenAIRE

    Owens, D R; Vora, J P; Birtwell, J; Luzio, S; Hayes, T M

    1988-01-01

    1. The absorption of 6U of soluble human insulin following subcutaneous injection into the anterior abdominal wall, thigh and into the thigh following admixture with aprotinin was assessed in normal subjects. The plasma immunoreactive insulin profiles were determined during a 6 h post injection period in subjects receiving concomitantly somatostatin to suppress endogenous insulin secretion. 2. Subcutaneous injection of human insulin into the anterior abdominal wall compared with the thigh led...

  17. Assessment of Aprotinin Loaded Microemulsion Formulations for Parenteral Drug Delivery: Preparation, Characterization, in vitro Release and Cytotoxicity Studies.

    Science.gov (United States)

    Okur, Neslihan Üstündağ; Özdemir, Derya İlem; Kahyaoğlu, Şennur Görgülü; Şenyiğit, Zeynep Ay; Aşıkoğlu, Makbule; Genç, Lütfi; Karasulu, H Yeşim

    2015-01-01

    The object of the current study was to prepare novel microemulsion formulations of aprotinin for parenteral delivery and to compare in vitro characteristics and release behaviour of different Technetium-99m ((99m)Tc)-Aprotinin loaded microemulsion formulations. In addition, cytotoxicity of microemulsion formulation was evaluated with cell culture studies on human immortalized pancreatic duct epithelial-like cells. For this aim, firstly, pseudo-ternary phase diagrams were plotted to detect the formulation region and optimal microemulsions were characterized for their thermodynamic stability, conductivity, particle size, zeta potential, viscosity, pH and in vitro release properties. For in vitro release studies aprotinin was labelled with (99m)Tc and labelling efficiency, radiochemical purity and stability of the radiolabeled complex were determined by several chromatography techniques. Radiolabeling efficiency of (99m)Tc-Aprotinin was found over than 90% without any significant changes up to 6 hours after labelling at room temperature. After that, in vitro release studies of (99m)Tc-Aprotinin loaded microemulsions were performed with two different methods; dissolution from diffusion cells and dialysis bags. Both methods showed that release rate of (99m)Tc- Aprotinin from microemulsion could be controlled by microemulsion formulations. Drug release from the optimized microemulsion formulations was found lower compared to drug solution at the end of six hours. According to stability studies, the optimized formulation was found to be stable over a period of 12 months. Also, human immortalized pancreatic duct epithelial-like cells were used to evaluate the cytotoxicity of optimum formulation. Developed microemulsion did not reveal cytotoxicity. In conclusion the present study indicated that the M1-APT microemulsion is appropriate for intravenous application of aprotinin. PMID:26306401

  18. 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

  19. 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

  20. Efficacy of tranexamic acid as compared to aprotinin in open heart surgery in children

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    Nagarajan Muthialu

    2015-01-01

    Full Text Available Background: Coagulopathy is a major issue in children undergoing high-risk pediatric cardiac surgery. Use of anti-fibrinolytics is well documented in adults, but recently there are questions raised about safety and effectiveness of their use on routine use. Tranexamic acid is a potent anti-fibrinolytic, but its role is not fully understood in children. This study aims to study the benefits tranexamic acid in controlling postoperative bleeding in pediatric cardiac surgical patients. Methods and Results: Fifty consecutive children who underwent cardiac surgery were randomized prospectively to receive either aprotinin (Group A; n = 24 or tranexamic acid (Group B; n = 26 from September 2009 to February 2010 were studied. Primary end points were early mortality, postoperative drainage, reoperation for bleeding and complications. Mean age and body weight was smaller in Group A (Age: 48.55 vs. 64.73 months; weight 10.75 vs. 14.80 kg respectively. Group A had more cyanotic heart disease than Group B (87.5% vs. 76.92%. Mean cardiopulmonary bypass time (144.33 vs. 84.34 min and aortic cross-clamp time (78.5 vs. 41.46 min were significantly higher in group A. While the blood and products usage was significantly higher in Group A, there was no difference in indexed postoperative drainage in first 4, 8 and 12 h and postoperative coagulation parameters. Mean C-reactive protein was less in Group A than B and renal dysfunction was seen more in Group A (25% vs. 7.6%. Mortality in Group A was 16.66% and 7.6% in Group B. Conclusion: Anti-fibrinolytics have a definitive role in high-risk children who undergo open-heart surgery. Tranexamic acid is as equally effective as aprotinin with no additional increase in morbidity or mortality. Ultramini Abstract: Coagulopathy has been a major issue in pediatric cardiac surgery, and anti-fibrinolytics have been used fairly regularly in various settings. This study aims to evaluate the efficacy of tranexamic acid as compared

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

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    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.

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

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    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

  3. 抑肽酶在体外循环心脏手术中的局部应用%The Topical Use of Aprotinin in Cardiac Surgery with Cardiopulmonary Bypass

    Institute of Scientific and Technical Information of China (English)

    陈亦江; 王晓伟; 陈广明

    2003-01-01

    Objective To investigate the effects of the topical use of aprotinin an the basisof comprrehensive blood conservations in cardiopulmonary bypass ( CPB). Methods In a prospectiveclinical trial, 20 patients were randomly divided into 2 groups. Control group: placebo was used topical-ly. Aprotinin group: aprotinin was poured into the pericardial cavity before closure of the sternotomy.Before and 24 h after surgery, hemoglobin ( Hb), hematocrit ( Hct), bleeding time (BT), clottingtime (CT) and prothrombin time (PT) were measured. Meanwhile, amounts of the mediastinaldrainage and the hemoglobin loss were observed at 0, 2, 6 and 24 h after operation. The samples fromthe mediastinal drainage were also collected to measure D-Dimer (D-D), tissue type plasminogen activa-tor (t-PA) activity, plasminogen activator inhibitor ( PAI) activity and protein C (PC). ResultsIn Aprotinin group, D-D, t-PA activity and PC were significantly reduced, compared with those in Con-trol group ( P < 0. 05, P<0. 05, P<0. 01). On the contrary, PAI activity was significantly in-creased, compared with that in Control group. Amounts of the mediastinal drainage and the hemoglobinloss were decreased by 43% and 52%, compared with those in Control group. Conclusion Ourresults suggest that the topical use of aprotinin can have better effects an the basis of comprehensive mod-erate blood conservation.%目的:探讨体外循环(CPB)心脏手术中采取综合性血液保护措施的基础上(转流中预充小剂量抑肽酶4~5万kIU/kg,手术失血回输,术后机器余血回输)再局部应用抑肽酶的血液保护效果.方法:体外循环心脏手术病人20例随机分为对照组(C组10例)、局部应用抑肽酶组(A组10例).检测指标:①D-二聚体(D-D)含量、组织型纤溶酶原激活物(t-PA)活性、组织型纤溶酶原激活物抑制剂(PAI)活性、蛋白C(PC)含量;②纵隔心包引流量、血红蛋白丢失.结果:局部用抑肽酶组引流液中D-D含量、t-PA活性

  4. 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

  5. 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

  6. Aprotinina não influencia troponina I, NTproBNP e função renal em crianças operadas com circulação extracorpórea High-dose aprotinin does not affect troponin I, N-Terminal pro-B-type natriuretic peptid and renal function in children submitted to surgical correction with extracorporeal circulation

    Directory of Open Access Journals (Sweden)

    Cesar Augusto Ferreira

    2009-12-01

    Full Text Available OBJETIVO: Avaliar se o uso de aprotinina em altas doses hemostáticas pode influenciar as funções miocárdicas, renais e metabólicas em crianças operadas com circulação extracorpórea (CEC. 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, a droga foi administrada antes e durante a CEC. As disfunções miocárdicas e multiorgânicas foram analisadas por marcadores clínicos e bioquímicos. Foram consideradas significantes as diferenças com POBJECTIVE: To evaluate if the use of hemostatic high-dose aprotinin seems influence to myocardial, renal and metabolic functions in children submitted to surgical correction with extracorporeal circulation (ECC. Material and Methods A prospective randomized study was conducted on children aged 30 days to 4 years submitted to correction of acyanogenic congenital heart disease with ECC and divided into two groups: Control (n=9 and Aprotinin (n=10. In the Aprotinin Group the drug was administered before and during ECC and the myocardial and multiorgan dysfunctions were analyzed on the basis of 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, permanence in the pediatric postoperative intensive care unit (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 24h after surgery in the Control Group. Blood loss was similar for both groups. Cardiac troponin I (cTnI, creatine kinase MB fraction (CKMB, serum

  7. 抑肽酶对体外循环呼吸指数和胸肺顺应性的影响%The influence of aprotinin on respiratory index and lung-thorax compliance during cardiopulmonary bypass

    Institute of Scientific and Technical Information of China (English)

    张兰; 王泉云; 肖红; 廖刃; 刘斌

    2001-01-01

    Objective To study the projection effect of aprotinin on acute lung injury during cardiopulmonary bypass (CPB). Methods Twenty-eight patients scheduled for cardiac value replacement for the first time were randomized into two groups: control group (14 patients) and aprotinin treatment group (14). Pulmonary thoracic compliance (Cs) and dynamic compliance (Cd) were measured at pre-CPB(T1), 10 minutes after aortic crossclamping (T2), and 10 minutes (T3), 60 minutes (T4) after CPB. Arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide(PaCO2), fractional concentration of oxygen in inspired gas (FiO2), and barometric pressure (PB) were measured at the same time. Then respiratory indexes (RI) were calculated. Results There were no differences in the values of Cs,Cd and RI at T1 between the aprotinin group(52.8±13.49, 42.97±9.06, 0.44±0.1) and the control group (56.43±9.29,45.62±6.92,0.45±0.13) respectively(P>0.05).There was a significant (P<0.05) decrease of Cs and Cd at T2 (33.82±8.83, 28.39±6.04), T3(35.42±11.83,30.6±17.88) and T4(37.96±7.27,32.38±6.52) in the control group as compared with basline values (T1)and the values of the aprotinin group (42.74±9.08,36.83±5.47; 43.38±10.88 , 37.27±10.57; 47.64±12.27, 40.52±9.07) respectively. The RI of the control group at T2 (0.78±0.14), T3(0.69±0.12) was higher than either their baseline(T1) or the values of the aprotinin group (0.53±0.14, 0.51±0.1) at the same measure points (P<0.05). Though there was no difference(P>0.05) between T4(0.63±0.13) and their baseline′s in the control group, the values of the control group were higher than those of the aprotinin group at T4(0.49±0.13)(P<0.05). Conclusion Aprotinin has a protection effect on acute lung injury after CPB.%目的探讨抑肽酶是否减轻体外循环(CPB)所致急性肺损伤。方法 28例首次心脏瓣膜置换术患者随机分为对照组及抑肽酶组,各14例。于CPB前(T1

  8. 氨甲环酸与抑肽酶对心脏手术血液保护的Meta分析%Effects of tranexamic acid and aprotinin on blood-conservation in heart surgery: a meta-analysis

    Institute of Scientific and Technical Information of China (English)

    叶博; 张国荣; 范勤; 曹晨; 魏国

    2011-01-01

    目的 系统评价氨甲环酸与抑肽酶在体外循环心脏手术中的血液保护效果.方法 检索Medline、PubMed、中国期刊全文数据库、中国生物医学文献数据库中氨甲环酸与抑肽酶对CPB心脏手术患者血液保护的随机对照研究文献,收集各研究中的血小板、凝血指标、血红蛋白、血细胞比容、术中出血量、术后24 h纵隔心包引流量、输血例数、术后并发症及术后近期死亡例数.计数资料采用优势比和95%可信区间表示,计量资料采用加权平均值和95%可信区间(CI)表示,所有计算和统计用RevMan 4.2.10软件完成,异质性检验采用χ2和I2完成.结果 符合标准的文献共16篇,3 393例患者.分析显示,与抑肽酶相比,氨甲环酸术后24 h纵隔心包引流量偏多(WMD=-65.28 CI:-103.92,-26.65 P=0.0009),但术后肾功能障碍患者例数明显较少(OR=1.36 CI:1.11,1.67 P=0.03),而血小板计数、凝血指标、血红蛋白值、术中出血量、成分输血例数均无显著差异(P>0.05).结论 在体外循环心脏手术中,氨甲环酸的血液保护效果比抑肽酶稍差,但其安全性能高,不易引起肾功能损害.%Objective To systematically evaluate the effect of tranexamic acid and aprotinin on blood conservation in open heart surgery with cardiopulmonary bypass.Methods A systematic overview and Meta - analysis were undertaken on all the randomized controlled trials of tranexamic acid and aprotinin in heart surgery from MEDLINE, PubMed, CNKI and CMB disk, with all the data as to platelets, coagulation indexes, hemoglobin ( Hb ), hematocrit ( Hct ), intraoperative hemorrhage volume, postoperative 24 h mediastinal pericardial drainage, cases of blood transfusion, postoperative complications and recent postoperative deaths.The weighted mean difference ( WMD ) with 95% confidence intervals ( CI ) for continuous data and odds ratio ( OR ) with 95% CI for dichotomous data were calculated.Statistical analysis was

  9. Gamma globulin, Evan's blue, aprotinin A PLA2 inhibitor, tetracycline and antioxidants protect epithelial cells against damage induced by synergism among streptococcal hemolysins, oxidants and proteinases: relation to the prevention of post-streptococcal sequelae and septic shock.

    Science.gov (United States)

    Ginsburg, I; Sadovnic, M

    1998-11-01

    An in vitro model was employed to study the potential role of streptococcal extra-cellular products, rich in streptolysin O, in cellular injury as related to streptococcal infections and post-streptococcal sequelae. Extra-cellular products (EXPA) rich in streptolysin O were isolated from type 4, group A hemolytic streptococci grown in a chemostat, in a synthetic medium. EXPA induced moderate cytopathogenic changes in monkey kidney epithelial cells and in rat heart cells pre-labeled with 3H-arachidonate. However very strong toxic effects were induced when EXP was combined with oxidants (glucose oxides generated H2O2, AAPH-induced peroxyl radical (ROO.), NO generated by sodium nitroprusside) and proteinases (plasmin, trypsin). Cell killing was distinctly synergistic in nature. Cell damage induced by the multi-component cocktails was strongly inhibited either by micromolar amounts of gamma globulin, and Evan's blue which neutralized SLO activity, by tetracycline, trasylol (aprotinin), epsilon amino caproic acid and by soybean trypsin inhibitor, all proteinase inhibitors as well as by a non-penetrating PLA2 inhibitor A. The results suggest that fasciitis, myositis and sepsis resulting from infections with hemolytic streptococci might be caused by a coordinated 'cross-talk' among microbial, leukocyte and additional host-derived pro-inflammatory agents. Since attempts to prolong lives of septic patients by the exclusive administration of single antagonists invariably failed, it is proposed that the administration of 'cocktails' of putative inhibitors against major pro-inflammatory agonizes generated in inflammation and infection might protect against the deleterious effects caused by the biochemical and pharmacological cascades which are known to be activated in sepsis. PMID:9848686

  10. 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.

  11. Fibrinolytic Inhibitors in Off-pump Coronary Surgery: A Prospective, Randomized, Double-Blind TAP Study (Tranexamic Acid, Aprotinin, Placebo)

    Czech Academy of Sciences Publication Activity Database

    Vaněk, T.; Jareš, M.; Fajt, R.; Straka, Z.; Jirásek, K.; Kolesár, M.; Brůček, P.; Malý, Marek

    2005-01-01

    Roč. 28, č. 4 (2005), s. 563-568. ISSN 1010-7940 Source of funding: V - iné verejné zdroje Keywords : tranexamic acid * protinin * off-pump coronary artery bypass * hemostasis Subject RIV: BB - Applied Statistics, Operational Research Impact factor: 1.802, year: 2005

  12. 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.

  13. 术中抑肽酶引起变态反应及处理%The treatment of anaphylactic or anaphylactoid reaction caused by intravenous aprotinin during surgical operation

    Institute of Scientific and Technical Information of China (English)

    林智平

    2002-01-01

    1 病例介绍 例1.男性,53岁,62 kg.ASAⅠ,行胃癌根治术.手术开始后静推112单位抑肽酶,约5 min,血压(mmHg)由给药前的110/76降至70/50,心率无明显增减,静注麻黄素20 mg,氢化可的松100 mg,非那根25 mg,快速输入胶体液5 min后未见明显改善,静滴间羟胺(100 mg加入5%葡萄糖盐水250 ml)后血压升至110/80.随后逐渐减慢静脉滴注间羟胺,15 min后撤药,输液速度恢复至正常速度,心率血压都维持在正常范围内直至手术结束,术后患者清醒,安全返回病室. 例2.女性,45岁,57 kg.ASAⅠ,无变态反应病史,行宫颈癌根治术.术前血压120/80,心率76 min-1.术中缓注抑肽酶112 U,2 min后心率降为38 min-1,血压降到56/30,立即静推肾上腺素0.5 mg(用5%葡萄糖盐水稀释到20 ml),非那根25 mg,氢化可的松100 mg,加快输入胶体,2 min后心率升至110 min-1,血压100/70,继续适当的快速静滴胶体、晶体直至手术结束.术后清醒,安全送返病室. 例3.女性,40岁,60 kg.ASAⅠ,无变态反应史,行乳腺癌根治术.术中用抑肽酶后出现上胸部、面部红斑,此时心率血压均在正常范围内,立即静注氢化可的松100 mg,非那根25 mg.红斑在5 min后褪去,心率血压在随后的手术过程中都正常.

  14. 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 ...... result of the adaptation. Determination of the level of mRNA encoding aprotinin and the plasmid copy number pointed to different mechanisms responsible for the decline in aprotinin yield in the different strains. (C) 2008 Elsevier B.V. All rights reserved....

  15. Estradiol receptor has proteolytic activity that is responsible for its own transformation.

    OpenAIRE

    Puca, G A; Abbondanza, C; Nigro, V; Armetta, I.; Medici, N; Molinari, A M

    1986-01-01

    We have investigated the effect of various protease inhibitors and substrates on the hormone- and temperature-dependent binding of partially purified estradiol-receptor complex to isolated nuclei. Only serine protease substrates and inhibitors significantly depressed estradiol receptor transformation. At 20 degrees C, we observed 50% inhibition with about 3 microM aprotinin or with 1.4 mM diisopropyl fluorophosphate. Aprotinin also blocked those size and charge modifications of receptor that ...

  16. Are antifibrinolytic drugs equivalent in reducing blood loss and transfusion in cardiac surgery? A meta-analysis of randomized head-to-head trials

    Directory of Open Access Journals (Sweden)

    Stokes Barrie J

    2005-07-01

    Full Text Available Abstract Background Aprotinin has been shown to be effective in reducing peri-operative blood loss and the need for re-operation due to continued bleeding in cardiac surgery. The lysine analogues tranexamic acid (TXA and epsilon aminocaproic acid (EACA are cheaper, but it is not known if they are as effective as aprotinin. Methods Studies were identified by searching electronic databases and bibliographies of published articles. Data from head-to-head trials were pooled using a conventional (Cochrane meta-analytic approach and a Bayesian approach which estimated the posterior probability of TXA and EACA being equivalent to aprotinin; we used as a non-inferiority boundary a 20% increase in the rates of transfusion or re-operation because of bleeding. Results Peri-operative blood loss was significantly greater with TXA and EACA than with aprotinin: weighted mean differences were 106 mls (95% CI 37 to 227 mls and 185 mls (95% CI 134 to 235 mls respectively. The pooled relative risks (RR of receiving an allogeneic red blood cell (RBC transfusion with TXA and EACA, compared with aprotinin, were 1.08 (95% CI 0.88 to 1.32 and 1.14 (95% CI 0.84 to 1.55 respectively. The equivalent Bayesian posterior mean relative risks were 1.15 (95% Bayesian Credible Interval [BCI] 0.90 to 1.68 and 1.21 (95% BCI 0.79 to 1.82 respectively. For transfusion, using a 20% non-inferiority boundary, the posterior probabilities of TXA and EACA being non-inferior to aprotinin were 0.82 and 0.76 respectively. For re-operation the Cochrane RR for TXA vs. aprotinin was 0.98 (95% CI 0.51 to 1.88, compared with a posterior mean Bayesian RR of 0.63 (95% BCI 0.16 to 1.46. The posterior probability of TXA being non-inferior to aprotinin was 0.92, but this was sensitive to the inclusion of one small trial. Conclusion The available data are conflicting regarding the equivalence of lysine analogues and aprotinin in reducing peri-operative bleeding, transfusion and the need for re

  17. Thyroid stimulating hormone stability in serum

    International Nuclear Information System (INIS)

    Thyroid stimulating Hormone (TSH) is a thermo labile peptide hormone. It is unstable in serum and rapidly degrades when exposed to ambient temperature (temp) for considerable time. The stability of TSH with regard to storage temp, duration and added preservative was evaluated for performing TSH essay, Venous blood was collected in 5 ml plain and aprotinin containing glass tubes from 37 individuals aged 15-56 years serum obtained was analysed for TSH at zero time value, then divided into 3 aliquots, sets with and without aprotinin. One set was kept at room temperature (RT), 2nd at 4 degree centigrade and 3rd at 20 degree centigrade TSH was measured after 24 and 72 hours for comparison to the zero time value of TSH. Significant decline in TSH was seen in the samples stored at RT for 72 hours. This effect was abolished when aprotinin, the protease inhibitor, was added to the samples, No significant difference from zero time value was noticed in the aprotinin-treated or untreated sera when kept at RT for 24 hours or when stored at 4 degree centigrade -20 degree centigrade for 72 hours. Thus we concluded that proper storage and addition of aprotinin may significantly reduce TSH degradation. (author)

  18. Estradiol receptor has proteolytic activity that is responsible for its own transformation.

    Science.gov (United States)

    Puca, G A; Abbondanza, C; Nigro, V; Armetta, I; Medici, N; Molinari, A M

    1986-08-01

    We have investigated the effect of various protease inhibitors and substrates on the hormone- and temperature-dependent binding of partially purified estradiol-receptor complex to isolated nuclei. Only serine protease substrates and inhibitors significantly depressed estradiol receptor transformation. At 20 degrees C, we observed 50% inhibition with about 3 microM aprotinin or with 1.4 mM diisopropyl fluorophosphate. Aprotinin also blocked those size and charge modifications of receptor that are characteristic of the transformation process. The estradiol receptor was able to bind to aprotinin-agarose only under transforming conditions; i.e., the interaction was hormone- and temperature-dependent and inhibited by molybdate. Diisopropyl fluorophosphate, a covalent reagent for serine esterases, competitively inhibited the binding and specifically eluted the estradiol-receptor complex that had been bound to aprotinin-agarose. These results indicate that estradiol receptor transformation is due to the effect of a serine protease and that the receptor itself is endowed with this catalytic activity, which is triggered by the steroid. PMID:2426695

  19. 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.

  20. Antifibrinolytics in cardiac surgery

    Directory of Open Access Journals (Sweden)

    Achal Dhir

    2013-01-01

    Full Text Available Cardiac surgery exerts a significant strain on the blood bank services and is a model example in which a multi-modal blood-conservation strategy is recommended. Significant bleeding during cardiac surgery, enough to cause re-exploration and/or blood transfusion, increases morbidity and mortality. Hyper-fibrinolysis is one of the important contributors to increased bleeding. This knowledge has led to the use of anti-fibrinolytic agents especially in procedures performed under cardiopulmonary bypass. Nothing has been more controversial in recent times than the aprotinin controversy. Since the withdrawal of aprotinin from the world market, the choice of antifibrinolytic agents has been limited to lysine analogues either tranexamic acid (TA or epsilon amino caproic acid (EACA. While proponents of aprotinin still argue against its non-availability. Health Canada has approved its use, albeit under very strict regulations. Antifibrinolytic agents are not without side effects and act like double-edged swords, the stronger the anti-fibrinolytic activity, the more serious the side effects. Aprotinin is the strongest in reducing blood loss, blood transfusion, and possibly, return to the operating room after cardiac surgery. EACA is the least effective, while TA is somewhere in between. Additionally, aprotinin has been implicated in increased mortality and maximum side effects. TA has been shown to increase seizure activity, whereas, EACA seems to have the least side effects. Apparently, these agents do not differentiate between pathological and physiological fibrinolysis and prevent all forms of fibrinolysis leading to possible thrombotic side effects. It would seem prudent to select the right agent knowing its risk-benefit profile for a given patient, under the given circumstances.

  1. Demonstration of a neurotrophic factor for the maintenance of acetylcholinesterase and butyrylcholinesterase in the preganglionically denervated superior cervical ganglion of the cat.

    OpenAIRE

    Koelle, G B; Ruch, G A

    1983-01-01

    Under sodium pentobarbital anesthesia, the superior cervical ganglia of cats were preganglionically denervated bilaterally. The following day cats were reanesthetized, the external carotid and lingual arteries were ligated bilaterally, and the right common carotid artery was infused for 24 hr with an extract prepared from cat brain, spinal cord, and sciatic nerves, with and without the incorporation of aprotinin, an inhibitor of proteases. They were sacrificed 48 hr after denervation, and the...

  2. Present status and upcoming prospects of hedgehog pathway inhibitors in small cell lung cancer therapy

    OpenAIRE

    Naqvi, Syed Hassan Abbas; Naqvi, Syed Hassan Shiraz; Bandukda, Muhammad Yasin; Naqvi, Syed Mumtaz Ali

    2013-01-01

    Lung cancer is an important etiology of malignant mortality worldwide with global statistics indicating over 1 million deaths annually. Although there have been advances in cytotoxic chemotherapy, the prognosis after treatment still remains poor. Remarkably, recent studies on the molecular level are creating the possibility to hamper lung cancer by inhibiting the hedgehog pathway. Currently, hedgehog pathway inhibitors include IWP-2, cyclopamine and aprotinin. However, Vismodegib is a new upc...

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

    OpenAIRE

    WANG, Meng; Zheng, Xin-Feng; Jiang, Lei-Sheng

    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...

  4. Cholinesterase activity in rat liver and serum during experimentally induced inflammation.

    Science.gov (United States)

    Simon, G; Budavári, I

    1977-01-01

    Cholinesterase activity of albino rats with acute local oedematous inflammation induced by turpentine, croton oil or Freund's adjuvant was elevated in the liver homogenate but decreased in the serum. Aprotinin administration prevented the decrease of serum activity. In the oedema fluid of rats treated with croton oil an enzyme with cholinester splitting activity was detected and it was shown to be identical with serum cholinesterase (EC 3. 1. 1. 8.). PMID:311577

  5. Role of the endogenous kallikrein-kinin system in modulating vasopressin-stimulated water flow and urea permeability in the toad urinary bladder.

    OpenAIRE

    Carvounis, C P; Carvounis, G; Arbeit, L A

    1981-01-01

    This study investigates the endogenous kallikrein-kinin system's role as a modulator of vasopressin action in the toad urinary bladder. Kalli-krein inhibition by aprotinin, which results in decreased kinin production, significantly increased both vasopressin and 8-Br-cyclic (c) AMP-stimulated water flow. Kinin potentiation by the kininase II inhibitor captopril (SQ 14225) significantly decreased vasopressin and 8-Br-cAMP-stimulated water flow. In contrast to water flow, vasopressin-stimulated...

  6. Potential effects of PKC or protease inhibitors on acute pancreatitis-induced tissue injury in rats.

    OpenAIRE

    Shi, Changbin; Zhao, Xia; Wang, Xiangdong; Zhao, Liming; Andersson, Roland

    2007-01-01

    Background: Acute pancreatitis (AP) is still one of the severe diseases, that cause the development of multiple organ dysfunction with a high mortality. Effective therapies for AP are still limited, mainly due to unclear mechanisms by which A-P initiates both pancreatic and extrapancreatic organ injury. Methods: Protease inhibitors (aprotinin, pefabloc, trypsin inhibitor) and PKC inhibitors (polymyxin B, staurosporine) were administrated 30 min before 'induction of AP in rats. To investig...

  7. Measurement of trypsin in duodenal juice by radioimmunoassay.

    OpenAIRE

    Lake-Bakaar, G; McKavanagh, S; Rubio, C E; Epstein, O; Summerfield, J A

    1980-01-01

    Trypsin in duodenal aspirate and pure pancreatic juice samples has been measured by both radioimmunoassay and enzymatic mathods. The radioimmunossay has been shown to be specific and to detect trypsin in the presence of aprotinin (Trasylol). In duodenal juice samples from control subjects and from patients with primary biliary cirrhosis a good correlation was obtained between both immunoreactive trypsin concentration and trypsin activity. The immunoreactive trypsinogen concentration in pure p...

  8. Identification and characterization of hydrolytic enzymes from the midgut of the cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

    OpenAIRE

    Özgür, Ebru; Yücel, Meral; Öktem, Hüseyin Avni

    2009-01-01

    Midgut hydrolytic enzymes of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) were identified and partially characterized. Km, Vmax, optimum pH, and specific activity were determined for proteolytic enzymes and a-amylases. All hydrolytic enzyme activity had an optimum pH value in the alkaline pH range. We observed major serine protease activity, together with minor cysteine-like activity, the former being significantly inhibited by soybean trypsin inhibitor (SBTI) and aprotinin. Moreover,...

  9. Antifibrinolytics in liver surgery

    OpenAIRE

    Jalpa Makwana; Saloni Paranjape; Jyotsna Goswami

    2010-01-01

    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 indica...

  10. Leishmania donovani secretory serine protease alters macrophage inflammatory response via COX-2 mediated PGE-2 production.

    Science.gov (United States)

    Das, Partha; De, Tripti; Chakraborti, Tapati

    2014-12-01

    Leishmania parasites determine the outcome of the infection by inducing inflammatory response that suppresses macrophage's activation. Defense against Leishmania is dependent on Th1 inflammatory response by turning off macrophages' microbicidal property by upregulation of COX-2, as well as immunosuppressive PGE-2 production. To understand the role of L. donovani secretory serine protease (pSP) in these phenomena, pSP was inhibited by its antibody and serine protease inhibitor, aprotinin. Western blot and TAME assay demonstrated that pSP antibody and aprotinin significantly inhibited protease activity in the live Leishmania cells and reduced infection index of L. donovani-infected macrophages. Additionally, ELISA and RT-PCR analysis showed that treatment with pSP antibody or aprotinin hold back COX-2-mediated immunosuppressive PGE-2 secretion with enhancement of Th1 cytokine like IL-12 expression. This was also supported in Griess test and NBT assay, where inhibition of pSP with its inhibitors elevated ROS and NO production. Overall, our study implies the pSP is involved in down-regulation of macrophage microbicidal activity by inducing host inflammatory responses in terms of COX-2-mediated PGE-2 release with diminished reactive oxygen species generation and thus suggests its importance as a novel drug target of visceral leishmaniasis. PMID:25823228

  11. Induction of DNA synthesis in isolated nuclei by cytoplasmic factors: inhibition by protease inhibitors

    International Nuclear Information System (INIS)

    Cytoplasmic extracts from spontaneously proliferating and mitogen-activated lymphoid cells contain a protein factor called ADR (activator of DNA replication) that induces DNA synthesis in isolated quiescent nuclei. ADR-containing preparations have proteolytic activity, as indicated by their ability to degrade fibrin in a plasminogen-independent and plasminogen-dependent manner. In addition, aprotinin, a nonspecific protease inhibitor, abrogates ADR-induced DNA synthesis in a dose-dependent fashion. Preincubation studies demonstrated that the effect of aprotinin is not due to its suppressive effects on the nuclei themselves. Other protease inhibitors such as leupeptin, p-aminobenzamidine, and N-α-tosyllysine chloromethyl ketone are also inhibitory, but soybean trypsin inhibitor is without effect. ADR activity can be removed from active extracts by adsorption with aprotinin-conjugated agarose beads and can be recovered by elution with an acetate buffer (pH 5). These finding are consistent with the interpretation that the initiation of DNA synthesis in resting nuclei may be protease dependent and, further, that the cytoplasmic stimulatory factor the authors have called ADR may be a protease itself

  12. A genetically engineered human Kunitz protease inhibitor with increased kallikrein inhibition in an ovine model of cardiopulmonary bypass.

    Science.gov (United States)

    Ohri, S K; Parratt, R; White, T; Becket, J; Brannan, J J; Hunt, B J; Taylor, K M

    2001-05-01

    A recombinant human serine protease inhibitor known as Kunitz protease inhibitor (KPI) wild type has functional similarities to the bovine Kunitz inhibitor, aprotinin, and had shown a potential to reduce bleeding in an ovine model of cardiopulmonary bypass (CPB). The aim of this study was to assess KPI-185, a modification of KPI-wild type that differs from KPI-wild type in two amino acid residues and which enhances anti-kallikrein activity in a further double-blind, randomized study in an ovine model of CPB, and to compare with our previous study of KPI-wild type and aprotinin in the same ovine model. Post-operative drain losses and subjective assessment of wound 'dryness' showed no significant differences between KPI-185 and KPI-wild type, despite the significant enhancement of kallikrein inhibition using KPI-185 seen in serial kallikrein inhibition assays. These preliminary findings support the hypothesis that kallikrein inhibition is not the major mechanism by which Kunitz inhibitors such as aprotinin reduce perioperative bleeding. PMID:11419655

  13. 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. PMID:27463548

  14. Capillary electrophoresis - electrospray ionization mass spectrometry in small diameter capillaries

    Energy Technology Data Exchange (ETDEWEB)

    Wahl, J.H.; Goodlett, D.R.; Udseth, H.R.; Smith, R.D.

    1992-06-01

    Methods (such as small inner diameter capillaries) are being explored to increase analyte sensitivity in capillary electrophoresis- electrospray ionization/mass spectroscopy(CE-ESI/MS). Results are reported for melittin in a protein mixture, with 10 to 100 {mu}m ID capillaries; and for a mixture of aprotinin, cytochrome c, myoglobin, and carbonic anhydrase, with 5 to 50 {mu}m ID capillaries. It is shown that an increase in solute sensitivity occurs when small ID capillaries ({lt} 20 {mu}m) are used in CE-ESI/MS for both a peptide and a protein mixture. 3 figs. (DLC)

  15. [Experimental studies of the prevention of postoperative adhesions].

    Science.gov (United States)

    Schwarz, J; Donat, H

    1989-01-01

    Female wistar rats were used for the testing of different substances to prevent postoperative intraabdominal adhesions. It could be demonstrated that the best results were obtained by dextran 70 with a concentration of 10%. Good effects were seen also by contrykal (aprotinin) and hylase (hyaluronidase). By the combination of dextran 70 30% with contrykal the spread of adhesions was very low but the effect was not significant better than with dextran 70 10% alone. For the prevention of intraabdominal adhesions operative techniques with minimal lesions are important too of the peritoneal epithelium. PMID:2466384

  16. 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 as...... subsequent hormone analysis. Our data support addition of DPP-4 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...

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

    International Nuclear Information System (INIS)

    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

  18. 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

    Large amounts of epidermal growth factor (EGF) are excreted in urine and the majority of this urinary EGF appears to be of renal origin. EGF is synthesized in the kidneys as a membrane-bound 160 kDa precursor, in the thick ascending limb of Henle and in the early part of the distal convoluted tub....... This is further evidence suggesting an intrarenal source of urinary EGF and suggests that the EGF precursor in the rat kidney is processed by enzyme(s) of renal origin.......Large amounts of epidermal growth factor (EGF) are excreted in urine and the majority of this urinary EGF appears to be of renal origin. EGF is synthesized in the kidneys as a membrane-bound 160 kDa precursor, in the thick ascending limb of Henle and in the early part of the distal convoluted....... 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...

  19. Immobilization of enzymes using non-ionic colloidal liquid aphrons (CLAs): Activity kinetics, conformation, and energetics.

    Science.gov (United States)

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

    2016-05-01

    This study seeks to examine the ability of non-ionic/non-polar Colloidial Liquid Aphrons (CLAs) to preserve enzyme functionality upon immobilization and release. CLAs consisting of micron-sized oil droplets surrounded by a thin aqueous layer stabilized by a mixture of surfactants, were formulated by direct addition (pre-manufacture addition) using 1% Tween 80/mineral oil and 1% Tween 20 and the enzymes lipase, aprotinin and α-chymotrypsin. The results of activity assays for both lipase and α-chymotrypsin showed that kinetic activity increased upon immobilization by factors of 7 and 5.5, respectively, while aprotinin retained approximately 85% of its native activity. The conformation of the enzymes released through desorption showed no significant alterations compared to their native state. Changes in pH and temperature showed that optimum conditions did not change after immobilization, while analysis of activation energy for the immobilized enzyme showed an increase in activity at higher temperatures. Furthermore, the effect of bound water within the aphron structure allowed for some degree of enzyme hydration, and this hydration was needed for an active conformation with results showing a decrease in ΔH* for the immobilized system compared to its native counterpart. Biotechnol. Bioeng. 2016;113: 970-978. © 2015 Wiley Periodicals, Inc. PMID:26497856

  20. Structural Evidence for Regulation and Specificity of Flaviviral Proteases and Evolution of the Flaviviridae Fold

    Energy Technology Data Exchange (ETDEWEB)

    Aleshin,A.; Shiryaev, S.; Strongin, A.; Liddington, R.

    2007-01-01

    Pathogenic members of the flavivirus family, including West Nile Virus (WNV) and Dengue Virus (DV), are growing global threats for which there are no specific treatments. The two-component flaviviral enzyme NS2B-NS3 cleaves the viral polyprotein precursor within the host cell, a process that is required for viral replication. Here, we report the crystal structure of WNV NS2B-NS3pro both in a substrate-free form and in complex with the trypsin inhibitor aprotinin/BPTI. We show that aprotinin binds in a substrate-mimetic fashion in which the productive conformation of the protease is fully formed, providing evidence for an 'induced fit' mechanism of catalysis and allowing us to rationalize the distinct substrate specificities of WNV and DV proteases. We also show that the NS2B cofactor of WNV can adopt two very distinct conformations and that this is likely to be a general feature of flaviviral proteases, providing further opportunities for regulation. Finally, by comparing the flaviviral proteases with the more distantly related Hepatitis C virus, we provide insights into the evolution of the Flaviviridae fold. Our work should expedite the design of protease inhibitors to treat a range of flaviviral infections.

  1. 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.

  2. 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.……

  3. 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

    shows that the membrane fraction of homogenized rat kidney contains an enzyme that releases immuno and receptor reactive EGF from the kidney membranes when incubated at 37 degrees C. Gel filtration shows that the EGF reactivity released from the membranes is similar to the EGF reactivity in rat urine......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 study....... 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....

  4. Temperature-dependent dissociation of homologous in-vivo produced complexes of 125I-des-AB fibrin monomers

    International Nuclear Information System (INIS)

    Des-AB fibrin monomers are prepared by incubation of fibrinogen with thrombin in the presence of aprotinin and EDTA. Plasma samples of rabbits taken at fixed time intervals show an exponential decrease of radioactivity after incorporation of fibrin monomers prepared by means of 125-I-labelled fibrinogen. The fibrin monomer complexes have been separated from the dimers or the fibrinogen by agarose gel filtration for the determination of radioactivity decrease in the various fractions. The results show a specific curve of degradation of the des-AB fibrin monomer complexes with an interpolated half-time of 13.5 hours. The experiments furthermore showed that these soluble fibrin monomer complexes will dissociate into equal parts of complexes of lower molecular weight and monomeric fibrin subsequent to a temperature increase from 200C to 370C. It can be assumed that this dissociation process favourably affects the capability of intravascular fibrin to act as a solubility promoting agent. (orig.)

  5. Proteolytic activity of the purified hormone-binding subunit in the estrogen receptor.

    Science.gov (United States)

    Molinari, A M; Abbondanza, C; Armetta, I; Medici, N; Minucci, S; Moncharmont, B; Nigro, V; Puca, G A

    1991-05-15

    The hormone-binding subunit of the calf uterus estradiol receptor was purified as a hormone-free molecule. Immunoaffinity chromatography with a specific monoclonal antibody was used as the final step. The purified subunit was specifically labeled by radioactive diisopropyl fluorophosphate. The diisopropyl fluorophosphate-labeled amino acid was serine. The purified receptor was able to release the fluorogenic or chromogenic group from synthetic peptides containing phenylalanine at the carboxyl terminus. This occurred only in the presence of estradiol and was hampered by aprotinin and diisopropyl fluorophosphate. Estradiol-dependent hydrolytic activity was also found in the eluate from gel slices after SDS/PAGE of purified receptor. This activity comigrated with the renaturable estradiol-binding activity. The estradiol antagonists 4-hydroxytamoxifen and ICI 164,384 as well as other steroid hormones were unable to activate this hydrolytic activity. PMID:1709742

  6. Topology of the fibrinolytic system within the mural thrombus of human abdominal aortic aneurysms.

    Science.gov (United States)

    Houard, X; Rouzet, F; Touat, Z; Philippe, M; Dominguez, M; Fontaine, V; Sarda-Mantel, L; Meulemans, A; Le Guludec, D; Meilhac, O; Michel, J-B

    2007-05-01

    Development and progression of acquired abdominal aortic aneurysms (AAAs) involve proteolytic activity. In the present study, we investigate the distribution of fibrinolytic system components within mural thrombi of human AAAs. 20 mural thrombi and the remaining AAA walls were dissected. The luminal, intermediate and abluminal thrombus layers, and media and adventitia were separately incubated in cell culture medium. Conditioned media were then analysed for plasminogen activators (PAs), plasminogen activator inhibitor-1 (PAI-1), free-plasmin, plasmin alpha(2)-antiplasmin complexes (PAPs) and D-dimers release. In parallel, PA and PAI-1 mRNA expression analysis was performed by RT-PCR. The study was completed by immunohistochemical localization of these components in AAA, ex vivo functional imaging using (99m)Tc-aprotinin as a ligand and measurement of PAP and D-dimer plasma levels. All fibrinolytic system components were present in each aneurysmal layer. However, the mural thrombus was the main source of active serine-protease release. Interestingly, the luminal layer of the thrombus released greater amounts of PAPs and D-dimers. This paralleled the preferential immunolocalization of plasminogen and PAs, and the (99m)Tc-aprotinin scintigraphic signal observed in the luminal pole of the thrombus. In contrast, mRNA expression analysis showed an exclusive synthesis of tPA and PAI-1 within the wall, whereas uPA mRNA was also expressed within the thrombus. Taken together, these results suggest that the increased plasma concentrations of PAPs and D-dimers found in AAA patients are related to mural thrombus proteolytic activity, thus explaining their known link with AAA progression. Components of the fibrinolytic system could also represent a target for functional imaging of thrombus activities in AAA. PMID:17352452

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

    International Nuclear Information System (INIS)

    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

  8. Human breast cancer cell-mediated bone collagen degradation requires plasminogen activation and matrix metalloproteinase activity

    Directory of Open Access Journals (Sweden)

    Hill Peter A

    2005-02-01

    Full Text Available Abstract Background Breast cancer cells frequently metastasize to the skeleton and induce extensive bone destruction. Cancer cells produce proteinases, including matrix metalloproteinases (MMPs and the plasminogen activator system (PAS which promote invasion of extracellular matrices, but whether these proteinases degrade bone matrix is unclear. To characterize the role that breast cancer cell proteinases play in bone degradation we compared the effects of three human breast cancer cell lines, MDA-MB-231, ZR-75-1 and MCF-7 with those of a normal breast epithelial cell line, HME. The cell lines were cultured atop radiolabelled matrices of either mineralized or non-mineralized bone or type I collagen, the principal organic constituent of bone. Results The 3 breast cancer cell lines all produced significant degradation of the 3 collagenous extracellular matrices (ECMs whilst the normal breast cell line was without effect. Breast cancer cells displayed an absolute requirement for serum to dissolve collagen. Degradation of collagen was abolished in plasminogen-depleted serum and could be restored by the addition of exogenous plasminogen. Localization of plasmin activity to the cell surface was critical for the degradation process as aprotinin, but not α2 antiplasmin, prevented collagen dissolution. During ECM degradation breast cancer cell lines expressed urokinase-type plasminogen activator (u-PA and uPA receptor, and MMPs-1, -3, -9,-13, and -14. The normal breast epithelial cell line expressed low levels of MMPs-1, and -3, uPA and uPA receptor. Inhibitors of both the PAS (aprotinin and PA inhibitor-1 and MMPs (CT1166 and tisue inhibitor of metalloproteinase blocked collagen degradation, demonstrating the requirement of both plasminogen activation and MMP activity for degradation. The activation of MMP-13 in human breast cancer cells was prevented by plasminogen activator inhibitor-1 but not by tissue inhibitor of metalloproteinase-1, suggesting

  9. 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

  10. Labeling proteins with Tc-99m via hydrazinonicotinamide (HYNIC): optimization of the conjugation reaction

    International Nuclear Information System (INIS)

    At present there is considerable interest in labeling peptides with Tc-99m for the development of target specific radiopharmaceuticals for imaging purposes. In the present study the conjugation of the bifunctional coupling agent succinimidyl-hydrazinonicotinamide (S-HYNIC) was studied and optimized in a series of peptides [molecular weight (MW) 6.5-14.3 kDa]. Aprotinin (MW 6.5 kDa), cytochrome C (MW 12.4 kDa), α-lactalbumin (MW 14.2 kDa), and lysozyme (MW 14.3 kDa) were conjugated with S- via the ε amino groups of their lysine residues. The effects of molar conjugation ratio, reaction temperature, pH, and protein concentration were studied. Reaction products were analyzed both with respect to the HYNIC-substitution ratio (spectrophotometrically) as well as to the labeling efficiency silica gel-instant thin layer chromatography (SG-ITLC) and molecular size fast performance liquid chromatography (FPLC). The effects of conjugation on biological activity were studied in three proteins binding to receptors on leukocytes: interleukin-8 (MW 8.5 kDa), interleukin-1α (MW 17 kDa), and interleukin-1 receptor antagonist (MW 17 kDa). The labeling efficiency of aprotinin, cytochrome c, α-lactalbumin, and lysozyme conjugated under optimal conjugation conditions exceeded 90%. Specific activities obtained were up to 7.5 MBq/μg. Conjugation was most efficient at 0 deg. C (as compared to 20 and 40 deg. C), at pH 8.2 (as compared to 6.0, 7.2, and 9.5), and at protein concentrations ≥ 2.5 mg/mL. In general, efficiency increased with increasing molar conjugation ratio (protein-HYNIC-ratio 1:3 99mTc using S-HYNIC is easy, rapid, and efficient, and preparations with high specific activity can be obtained. However, biological activity of proteins may be lost at high HYNIC-substitution ratios. With the proteins tested here a careful balancing of reaction conditions resulted in acceptable, although suboptimal, labeling efficiencies and preservation of biological activity

  11. Optimization of fibrinolytic protease production from Bacillus subtilis I-2 using agro-residues

    Directory of Open Access Journals (Sweden)

    Bijender Kumar Bajaj

    2014-10-01

    Full Text Available The aim of this work was to study the production of fibrinolytic protease by Bacillus subtilis I-2 on agricultural residues. Molasses substantially enhanced (63% protease production (652.32 U/mL than control (398.64 U/mL. Soybean meal supported maximum protease production (797.28 U/mL, followed by malt extract (770.1 U/mL, cotton cake (761.04 U/mL, gelatin (742.92 U/mL and beef extract (724.8 U/mL. Based on the Plackett-Burman designed experiments, incubation time, soybean meal, mustard cake and molasses were identified as the significant fermentation parameters. Ammonium sulfate precipitation and DEAE sephadex chromatography resulted 4.8-fold purification of protease. Zymography showed the presence of three iso-forms in the partially purified protease preparation, which was confirmed by the SDS-PAGE analysis (42, 48, 60 kDa. Protease exhibited maximum activity at 50oC and at pH 8.0. Significant stability was observed at 30-50oC and at pH 7.0-10.0. Mg2+, Zn2+, Co2+, Ca2+, Mn2+ and Cu2+,EGTA, EDTA and aprotinin severely decreased the enzyme activity.

  12. Involvement of Kallikrein-Kinin System on Cardiopulmonary Alterations and Inflammatory Response Induced by Purified Aah I Toxin from Scorpion Venom.

    Science.gov (United States)

    Medjadba, Wafa; Martin-Eauclaire, Marie-France; Laraba-Djebari, Fatima

    2016-02-01

    Bradykinins are released from kininogen by kallikrein. They increase capillary lung permeability after their binding to β1 and especially β2 receptors before being metabolized by kininase enzyme. This study was performed to evaluate cardiopulmonary damages and inflammatory response on injected rats with Aah I toxin of scorpion venom and the involvement of Kallikrein-Kinin system in this pathogenesis. Obtained results revealed that Aah I toxin induces inflammatory cell infiltration accompanied by cellular peroxidase activities, a release of cytokine levels, pulmonary and myocardial damage, with altered metabolic activities and imbalanced redox status. Administration of aprotinin (bradykinin inhibitor) and especially icatibant (bradykinin β2 receptor antagonist) seemed to be able to protect animals against the toxicity of Aah I; nevertheless, the use of captopril (kininase II inhibitor) reduced partially some cardiac disorders. These findings indicate that the kallikrein-kinin system may contribute to the physiopathological effect and lung edema formation induced by toxin, which suggests a potential use of drugs with significant anti-kinin properties. PMID:26361946

  13. Iodine labelling of sea anemone toxin II, and binding to normal and denervated diaphragm

    International Nuclear Information System (INIS)

    1. Sea anemone toxin II (ATX II) which keeps the activated sodium channels open, can be labelled at its histidine residues with 125I up to a specific radioactivity of 500 Ci/mmole. Upon intraventricular injection in mice, ATX II causes acute, short-lasting hyperexcitation and convulsions. Its LD50 in mice is between 25 and 50 ng of the native peptide, and between 50 and 100 ng of the radioactive material per animal. 2. The labelled peptide is bound to mouse diaphragm from where it can be displaced by ATX II and, even better, by scorpion neurotoxin but not by other basic peptides, e.g., histone or aprotinin. Binding is not significantly influenced by 50 mM potassium, by replacing sodium with choline, by veratridine or tetrodotoxin. In contrast to binding of α-bungarotoxin, binding of ATX II is not changed by denervation of the diaphragm. ATX II binds not only to the muscular but also to the tendinous moiety of the mouse diaphragm. 3. ATX II lowers the surface tension of water. Further experiments are needed to establish the usefulness of 125I-ATX for labelling sodium channels in excitable membranes. (orig.)

  14. 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.

  15. 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. PMID:27468462

  16. Cloning, expression and characterisation of an HtrA-like serine protease produced in vivo by Mycobacterium leprae

    Directory of Open Access Journals (Sweden)

    Michelle Lopes Ribeiro-Guimarães

    2009-12-01

    Full Text Available Members of the high temperature requirement A (HtrA family of chaperone proteases have been shown to play a role in bacterial pathogenesis. In a recent report, we demonstrated that the gene ML0176, which codes for a predicted HtrA-like protease, a gene conserved in other species of mycobacteria, is transcribed by Mycobacterium leprae in human leprosy lesions. In the present study, the recombinant ML0176 protein was produced and its enzymatic properties investigated. M. lepraerecombinant ML0176 was able to hydrolyse a variety of synthetic and natural peptides. Similar to other HtrA proteins, this enzyme displayed maximum proteolytic activity at temperatures above 40°C and was completely inactivated by aprotinin, a protease inhibitor with high selectivity for serine proteases. Finally, analysis of M. leprae ML0176 specificity suggested a broader cleavage preference than that of previously described HtrAs homologues. In summary, we have identified an HtrA-like protease in M. lepraethat may constitute a potential new target for the development of novel prophylactic and/or therapeutic strategies against mycobacterial infections.

  17. 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

  18. Perioperative allergy: uncommon agents.

    Science.gov (United States)

    Caimmi, S; Caimmi, D; Cardinale, F; Indinnimeo, L; Crisafulli, G; Peroni, D G; Marseglia, G L

    2011-01-01

    Anesthesia may often be considered as a high-risk procedure and anaphylaxis remains a major cause of concern for anesthetists who routinely administer many potentially allergenic agents. Neuromuscular blocking agents, latex and antibiotics are the substances involved in most of the reported reactions. Besides these three agents, a wide variety of substances may cause an anaphylactic reaction during anesthesia. Basically all the administered drugs or substances may be potential causes of anaphylaxis. Among them, those reported the most in literature include hypnotics, opioids, local anesthetics, colloids, dye, Non-Steroidal Anti-Inflammatory Drugs (NSAIDs), Iodinated Contrast Media (ICM), antiseptics, aprotinin, ethylene oxyde and formaldehyde, and protamine and heparins. No premedication can effectively prevent an allergic reaction and a systematic preoperative screening is not justified for all patients; nevertheless, an allergy specialist should evaluate those patients with a history of anesthesia-related allergy. Patients must be fully informed of investigation results, and advised to provide a detailed report prior to future anesthesia. PMID:22014927

  19. Pharmacological characterization of rat paw edema induced by Cerastes gasperettii (cerastes venom

    Directory of Open Access Journals (Sweden)

    A. K. Al-Asmari

    2006-01-01

    Full Text Available Inflammatory response induced by the venom of the Arabian sand viper Cerastes gasperettii was studied by measuring rat hind-paw edema. Cerastes gasperettii venom (CgV, 3.75-240 µg/paw, heated for 30s at 97°C, caused a marked dose and time-dependent edema in rat paw. Response was maximal 2h after venom administration and ceased within 24h. Heated CgV was routinely used in our experiments at the dose of 120 µg/paw. Among all the drugs and antivenoms tested, cyproheptadine and 5-nitroindazole were the most effective in inhibiting edema formation. Aprotinin, mepyramine, dexamethasone, diclofenac, dipyridamole, Nomega-nitro-L-arginine, quinacrine, and nordihydroguaiaretic acid showed statistically (p<0.001 significant inhibitory effect, but with variations in their inhibition degree. Equine polyspecific and rabbit monospecific antivenoms significantly (p<0.001 reduced edema when locally administered (subplantar but were ineffective when intravenously injected. We can conclude that the principal inflammatory mediators were serotonin, histamine, adenosine transport factors, phosphodiesterase (PDE, cyclooxygenase, lipoxygenase and phospholipase A2 (PLA2, in addition to other prostaglandins and cytokines.

  20. 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. PMID:27280436

  1. Human urinary kallikrein (HUK): large-scale purification and direct solid-phase radioimmunoassay (RIA).

    Science.gov (United States)

    Morichi, S; Sako, E; Iga, Y; Nishida, M; Moriya, H

    1986-01-01

    HUK was purified from 1,000 liters of fresh urine by the following procedures: silica gel adsorption, gel filtration on Sephadex G-75, DEAE-Sephadex chromatography, bentonite treatment, affinity chromatography on aprotinin-Sepharose 4B, and rapid gel filtration on a TSK Gel G-3000 SWG column. Seventeen mg of HUK being found to be pure by means of various analyses was obtained. The pI values of the heterogeneous components of HUK were 3.5, 3.8, and 4.1, while the corresponding molecular weights of these components were 5.4 X 10(4), 4.9 X 10(4), and 4.4 X 10(4), respectively. The antigens (125I- and non-labeled HUK) were incubated for 4 hrs at 37 degrees C in polystyrene test tubes to which anti-HUK rabbit IgG had been immobilized. The quantitative range of the standard curve was 1-128 ng. This Ria principally recognized active form of HUK. Therefore, total and inactive HUK also could be determined by the combination of the RIA with trypsin treatment of the urine sample. The RIA correlated closely with both S-2266 amidolytic assay and kininogenase assay. PMID:3643743

  2. Plasmin: indigenous milk proteinase

    Directory of Open Access Journals (Sweden)

    Samir Kalit

    2002-06-01

    Full Text Available The most important characteristic of plasmin, as significant indigenous milk proteinase, its concentration, concentration measuring procedure and activity of plasmin are described. The most important factors, which have an influence on concentration and plasmin activity in milk, are stage of lactation and mastitis (high somatic cell count – SCC. In high SCC milk indigenous proteinase activity increased, especially in plasmin and plasminogen system.Specific hydrolytic activity of plasmin during primary proteolysis of some casein fractions is described. ß-CN is most susceptible fraction, but αs1-CN and αs2-Cn are less susceptible to degradation by plasmin. Almost all fractions of κ-CN are resistant to degradation by plasmin. Activation of plasminogen to plasmin is very complex biochemical process influenced by activators and inhibitors in milk, and can be increased in high SCC milk. There are many various types of inhibitors in milk serum and ßlactoglobulin is the most important after its thermal denaturation. Addition of aprotinin and soybean tripsin inhibitors in milk inhibits plasmin activity. Most important characteristic of plasmin is its thermostability onpasteurisation and even sterilisation. Mechanism of thermal inactivation of plasmin with developing covalent disulphide interaction between molecule of plasmin and serum proteins (mostly ß-laktoglobulin is described. Thermosensitive inhibitors of plasminogen activators and inhibitors of plasmin are inactivated by short pasteurisation and therefore increase plasmin activity,while higher temperature and longer treatment time inactivate plasmin activity.

  3. Complement system and immunological mediators: Their involvements in the induced inflammatory process by Androctonus australis hector venom and its toxic components.

    Science.gov (United States)

    Bekkari, Nadjia; Martin-Eauclaire, Marie-France; Laraba-Djebari, Fatima

    2015-01-01

    Androctonus australis hector scorpion venom is well known by its high toxicity, it induces massive release of neurotransmitters that lead to pathophysiological disorders in cardiovascular, neuro-hormonal and immune systems. Previous studies have shown the relationship between the severity of scorpion envenoming and immune system activation. This study was assessed to investigate the involvement of complement system and inflammatory mediators after sublethal injection of Aah venom, its toxic fraction (FtoxG50) and its main toxins (AahI and AahII) into NMRI mice. The Activation complement system by the venom is also compared to that induced of lipopolysaccharides (LPS). Obtained results showed that seric complement system (CS) is activated by the venom and by its toxic components; this activation is more pronounced into liver tissue when toxic components (FtoxG50, AahI or AahII) are used. Increase of cytokine levels (IL1β, TNFα and ICAM) into hepatic tissue induced by AahI or AahII neurotoxins is correlated with tissue alterations. Aprotinin, a non specific inhibitor of complement system seems to be able to reduce CS consumption and to restore partially the induced tissue damage by venom. The mechanisms by which toxic fraction or LPS induced the activation of complement system seem to be different. Sensitivity of hepatic tissue is more pronounced after FtoxG50 injection; however lung tissue is more sensible to LPS than FoxG50. PMID:25921955

  4. The role of myoglobin degradation in nephrotoxicity after rhabdomyolysis.

    Science.gov (United States)

    Zorova, Ljubava D; Pevzner, Irina B; Chupyrkina, Anastasia A; Zorov, Savva D; Silachev, Denis N; Plotnikov, Egor Y; Zorov, Dmitry B

    2016-08-25

    The fate of myoglobin in renal cells was explored in an animal model of rhabdomyolysis known as the pathology highly related to oxidative stress resulting in impairment of renal functioning. The working hypothesis was that the proper degradation of myoglobin in rhabdomyolytic kidney can activate the reparative processes in the tissue. We found that incubation of myoglobin with kidney cells causes its accumulation in the cytoplasm. In rhabdomyolytic rats, the level of heme and free iron in cytoplasm and mitochondria of kidney cells is remarkably increased while inhibition of proteolysis results in further elevation of myoglobin content in the renal tissue. Heme oxygenase and ferritin levels were found to be increased in the kidney tissue at rhabdomyolysis and simulating conditions performed by i/v injection of myoglobin. In addition, the level of peroxidized lipids was high in rhabdomyolytic kidney and became even higher after inhibition of proteolysis by aprotinin. Elevated levels of carbonylated proteins were also observed after rhabdomyolysis, however, if prior to induction of rhabdomyolysis the injection of myoglobin was done, the level of carbonylated proteins dropped versus unprimed kidney tissue thus affording protection to the kidney against oxidative stress. Injection of myoglobin to the rat results in impairment of renal functioning and inhibition of myoglobin degradation in the rhabdomyolytic animal aggravates acute renal failure, demonstrating that degradation of myoglobin is somehow beneficial although it may result in undesired release of free iron which can participate in toxic redox cycling. PMID:27329933

  5. C-peptide comparative radioimmunoassays: a study of three commercial kits

    International Nuclear Information System (INIS)

    Plasma C-peptide immunoreactivity (CPR) was measured in 18 fasting subjects with three different commercial kits (RIA-mat C-peptide-, Byk-Mallinckrodt; RIA-gnost-hC-peptide, Hoechst-Behring; human C-peptide radioimmunoassay kit, Novo) The subjects were chosen as to cover a wide range of CPR concentrations (five healthy subjects, six obese subjects, three insulin-dependent diabetics, four normal subjects whose plasmas had been kept at - 200C for periods of 16 or 36 months). CPR was measured with the Novo kit in eight other plasmas which were kept over a period of 36 months, with or without aprotinin. Good correlations have been established among the values found with the three kits. However, absolute concentration values for each subject as well as the dispersion of all plasma C-peptide values varied as a function of the kit used because of antibody specificity differences and because of the various separation methods. The normal range proposed changes with each kit and the blood CPR of a subject can be a normal, reduced or increased one, depending on the kit used. After several months of storage, plasma CPR degradation is observed with the three kits. A protease-inhibitor is necessary in order to avoid this C-peptide degradation due to the apparent existence of a plasma proteolytic enzyme

  6. 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.

  7. Isolation and characterization of human membrane carboxypeptidase (HMCP)

    International Nuclear Information System (INIS)

    The authors detected a membrane-bound carboxypeptidase in human placenta and other tissues which cleaves C-terminal Lys or Arg of peptides such as Lys6-Met5-enkephalin. The enzyme was solubilized from placental microvilli with 0.8% CHAPS and purified 427-fold by ion-exchange chromatography, Sepharose-arginine affinity chromatography, chromatofocusing and gel filtration on HPLC. HMCP had a mol. wt. of 67,000 in SDS-PAGE and 65,300 in gel filtration and a pH optimum of 7.0. HMCP cleaved Bz-Gly-argininic acid the fastest (90 μmol/min/mg) followed by Bz-Ala-Lys (41), Bz-Phe-Lys (26), Bz-Gly-Arg (1.7) and Bz-Gly-Lys (1.6). Activity was stimulated by CoCl2 and inhibited by cadmium acet., o-phenanthroline and 2-mercaptomethyl-3-guanidinoethylthiopropanoic acid but not by phenylmethylsulfonyl fluoride, aprotinin or p-chloromercuriphenylsulfonate. The enzyme was stable for 1 hr at room temp. at pH 4.25, but lost 31% activity at pH 4.0. HMCP did not react with antiserum to human plasma carboxypeptidase N in Western blotting. This study shows that human placental microvilli contain a membrane carboxypeptidase, that differs from other carboxypeptidases, and cleaves C-terminal basic amino acids from peptides. This enzyme could be involved in regulating the level of peptide hormones in the placenta and other tissues

  8. 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.

  9. Active site conformational changes of prostasin provide a new mechanism of protease regulation by divalent cations

    International Nuclear Information System (INIS)

    Prostasin or human channel-activating protease 1 has been reported to play a critical role in the regulation of extracellular sodium ion transport via its activation of the epithelial cell sodium channel. Here, the structure of the extracellular portion of the membrane associated serine protease has been solved to high resolution in complex with a nonselective d-FFR chloromethyl ketone inhibitor, in an apo form, in a form where the apo crystal has been soaked with the covalent inhibitor camostat and in complex with the protein inhibitor aprotinin. It was also crystallized in the presence of the divalent cation Ca+2. Comparison of the structures with each other and with other members of the trypsin-like serine protease family reveals unique structural features of prostasin and a large degree of conformational variation within specificity determining loops. Of particular interest is the S1 subsite loop which opens and closes in response to basic residues or divalent ions, directly binding Ca+2 cations. This induced fit active site provides a new possible mode of regulation of trypsin-like proteases adapted in particular to extracellular regions with variable ionic concentrations such as the outer membrane layer of the epithelial cell.

  10. Neutral protease (T-Kase) in the inflammatory fluid of the rat which releases T-Kinin from T-Kininogen

    International Nuclear Information System (INIS)

    T-Kininogen, an acute phase protein, and thiol protease inhibitor, has been shown by the laboratory to be the major kininogen in normal rat plasma (70%). In rats with adjuvant arthritis, levels in blood rise to over 90% of the total kininogen. The authors have demonstrated the presence of free T-Kinin in the inflammatory fluid of carrageenin-injected rats. Their present study shows that a neutral protease (T-Kase) is present along with Cathepsin D in inflammatory fluids which may be the enzymes releasing T-Kinin from T-Kininogen. 24 hrs after a 2% carrageenin solution (4.0 ml.) was injected into a dorsum air pouch of the rat, the rats were sacrificed and 100 μl of the resulting inflammatory fluid was incubated with T-Kininogen at varying pH in the presence of o-phenanthroline. Released kinins were subjected to HPLC chromatography and radioimmunoassayed. The results indicated the presence of two enzymes which release T-Kinin. One was an acidic protease with a pH optimum at 3.5 which is completely inhibited by pepstatin and is probably cathepsin-D. A second enzyme has a pH optimum at 7.0 and is not inhibited by pepstatin, leupeptin, aprotinin and SBTI. The properties of the neutral T-Kase are under study

  11. Scorpion (Buthus tamulus venom toxicity on cardiopulmonary reflexes involves kinins via 5-HT3 receptor subtypes

    Directory of Open Access Journals (Sweden)

    S. BAGCHI

    2001-01-01

    Full Text Available The mechanisms underlying the action of Indian red scorpion Buthus tamulus (BT venom-induced augmentation of cardiopulmonary reflexes elicited by intravenous injection of 5-HT were examined in urethane anaesthetized rats. The 5-HT produced a concentration-dependent increase in time-response area of bradycardiac response, with the responses at submaximal concentrations shifted to the left after exposure to BT venom (20 µg/kg, IV. Aprotinin (6000 kallikrein inactivating unit, IV as such had no effect on 5-HT reflex responses (bradycardia, hypotension, and apnea, but blocked the venom-induced reflex augmentation. While ondansetron (10 µg/kg, IV completely blocked the 5-HT reflex responses, these reappeared partially after venom exposure (20 µg/kg. Exposure to bradykinin (50 µg/kg, IV for 30 min also augmented the 5-HT-induced reflex responses similar to venom. The bradykinin-induced augmentation was also blocked by ondansetron. Results indicate that the venom-induced augmentation of cardiopulmonary reflexes is mediated through kinins sensitizing 5-HT3 receptor subtypes.

  12. Purification and characterization of a hemorrhagic metalloproteinase from Bothrops lanceolatus (Fer-de-lance) snake venom.

    Science.gov (United States)

    Stroka, Alessandra; Donato, José L; Bon, Cassian; Hyslop, Stephen; de Araújo, Albetiza Lôbo

    2005-03-15

    Bothrops snake venoms contain metalloproteinases that contribute to the local effects seen after envenoming. In this work, a hemorrhagic metalloproteinase (BlaH1) was purified from the venom of the snake Bothrops lanceolatus by a combination of gel filtration, affinity (metal chelating) and hydrophobic interaction chromatographies. The hemorrhagin was homogeneous by SDS-PAGE and had a molecular mass of 28 kDa that was unaltered by treatment with beta-mercaptoethanol. BlaH1 gave a single band in immunoelectrophoresis and immunoblotting using commercial bothropic antivenom. BlaH1 had hemorrhagic, caseinolytic, fibrinogenolytic, collagenolytic and elastinolytic activities, but no phospholipase A(2) activity. The hemorrhagic and caseinolytic activities were inhibited by EDTA, indicating that they were metal ion-dependent. In contrast, aprotinin, benzamidine and PMSF did not affect these activities. The caseinolytic activity of BlaH1 had a pH optimum of 8.0 and was stable in solution at up to 40 degrees C; activity was completely lost at > or =70 degrees C. The hemorrhagic activity was neutralized by commercial bothropic antivenom. These properties suggest that this new hemorrhagin belongs to class P-I snake venom metalloproteinases. PMID:15733562

  13. 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. PMID:25649205

  14. 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

  15. Comparison of the effects of various absorption enhancers and enzyme inhibitors on buccal insulin delivery in vitro and in vivo%不同吸收促进剂及酶抑制剂对胰岛素 体内及体外口腔黏膜渗透性的影响

    Institute of Scientific and Technical Information of China (English)

    杨天智; 陈大兵; 张强

    2001-01-01

    Objective: To study the effects of variou s absorption enhancers and enzyme inhibitors on in sulin permeation in vitro and in vivo . Methods: The pene tration of insulin throu gh hamster and rabbit buccal membrane was investigated by measuring in vitro transbuccal flux. Buccal insulin absorption was estimated in vivo from th e cumula tive response of serum glucose concentrations and comparison was made with the results of subcutaneous experiments. Results: There was a statistically significant permeabil ity increas e of insulin over controls after co-administration with the SDCh, Brij78, SLS or lecithin, aprotinin, bacitracin, whereas 1-menthol or poloxamer were less e ffective. Buccal insulin efficacy in the absence of co-administration adjuvants was very low relative t o subcutaneous administration of insulin. When co-administered buccally with SD Ch, SLS, l ecithin or Brij78, Fr (relative pharmacological bioavailability) values were al l increased significantly. Conclusion: The present studies showed that with the most effective absorption adjuvants, buccal insulin was one-fifth to one-fourth as effective as subcutaneous insulin. Results of in vitro experiments were in agreement with the in vivo re sults with respect to the enhancement of these adjuvants.%目的:研究不同的吸收促进剂及酶抑制剂对胰岛素透过口腔黏膜的影响。方法:体外实验中,在不同吸收促进剂及酶抑制剂作用下,测定胰岛素透过仓鼠和家兔口腔黏膜的渗透系数;体内实验中,在胰岛素口腔喷雾剂中加入不同的吸收促进剂及酶抑制剂,考察大鼠经口腔喷入胰岛素后的血糖降低情况。结果:SDCh, Brij78, SLS以及lecithin可以显著增加胰岛素透过口腔黏膜的渗透系数,而aprotinin,bacitracin, 1-menth ol 以及 poloxamer的作用相对较小。胰岛素溶液中加入SDCh, Brij78, SLS以及lecithin 后,正常大鼠口腔喷雾给药,药理生物利用度都有显著的提高,而aprotinin

  16. 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.

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

    International Nuclear Information System (INIS)

    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 (PAR1), and by PAR1 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 PAR1-PKC-Src-MMP pathway and HB-EGF/AR shedding-dependent EGFR transactivation.

  18. Coronary artery bypass grafting (CABG) in patients with immune thrombocytopenia (ITP): a community hospital experience and review of the literature.

    Science.gov (United States)

    Jubelirer, Steven J; Mousa, Luay; Reddy, Usha; Mir, Mohsin; Welch, Christine A

    2011-01-01

    We reviewed the records of 51 patients with Immune Thrombocytopenia (ITP) who underwent Coronary Artery Bypass Grafting (CABG) at Charleston Area Medical Center between June 1992 and September 2005. There were 41 males and 10 females with a median age of 68 years (range 49-87). Four patients had a previous splenectomy, one of whom had it performed concomitantly with the CABG. Three patients were on chronic corticosteroids on admission. The median pump time was 114 minutes (range: 42-244 minutes). The median cross-clamp time was 62 minutes (range 22-192 minutes). The median total chest tube drainage postoperatively was 1,346 cc (range: 265-9875cc). The mean preoperative and 24 hour postoperative platelet count was 126,000 (range 58,000-323,000) and 99,000/mm3 (range: 27,000-194,000), respectively. Twenty-one (40%) patients received platelet transfusions. Platelets were given intraoperatively or postoperatively in all but two of those patients. The median number of units of platelets given was 10 (range: 6-52). Twenty-seven (53%) received packed red cells intraoperatively or postoperatively. The median number of red cells given was 2 (range: 1-34). Other hemostatic agents given intraoperatively/ postoperatively included aprotinin (8 patients), aminocaproic acid (10 patients), DDAVP (5 patients), and intravenous gammaglobulin (IgG) in 3 patients. Thirteen patients were given corticosteroids preoperatively with little improvement in platelet count. CABG may be successfully performed in ITP patients with moderate thrombocytopenia (> or = 50,000/mm3) using conventional therapies (e.g., transfusions, IV IgG, hematinics) without the need for preoperative splenectomy or prolongation of hospital stay. However, a prospective study on the ideal management of ITP patients undergoing CABG would be beneficial. PMID:22235705

  19. Kallikreins when activating bradykinin B2 receptor induce its redistribution on plasma membrane.

    Science.gov (United States)

    Hecquet, Claudie; Becker, Robert P; Tan, Fulong; Erdös, Ervin G

    2002-12-01

    The bradykinin (BK) B2 receptor (R) is directly activated by kallikreins and other serine proteases independent of BK release. Both the Galpha(i) and Galpha(q) proteins are involved, shown by the release of arachidonic acid and [Ca2+]i elevation. Site-directed mutagenesis of the receptor and the lack of heterogeneous desensitization of the human B2R by the BK and kallikrein emphasize among others the differences between activation by the proteases and the peptide. To characterize further the mechanism thereby kallikreins activate and desensitize the B2R we investigated the distribution of the human B2R tagged with the green fluorescent protein (B2-GFP(Ct)) on the plasma membrane of stably transfected Chinese hamster ovary (CHO) cells. We visualized the movement of B2-GFP(Ct) R with confocal fluorescence microscopy after activation by BK or a by serine protease. Continued exposure of the cells to BK led to B2R internalization within 15-20 min. Porcine pancreatic and human recombinant tissue kallikreins induced a rapid definite redistribution of receptors on the plasma membrane within 5 min, prior to internalization. These effects of kallikrein were blocked by the B2R antagonist HOE 140 and by the kallikrein inhibitor, aprotinin. The B2R was also activated by endoproteinases LysC and ArgC and trypsin, but these enzymes did not induce redistribution, only internalization. In control experiments, kallikrein had no effect on cells transfected to stably express the angiotensin-converting enzyme-green fluorescent protein (GFP). Thus, kallikreins when activating the BK B2R also trigger its redistribution on plasma membrane. PMID:12489794

  20. 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.

  1. New options in the management of tendinopathy

    Directory of Open Access Journals (Sweden)

    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

  2. Time-dependent cellular morphogenesis and matrix stiffening in proteolytically responsive hydrogels.

    Science.gov (United States)

    Kesselman, Dafna; Kossover, Olga; Mironi-Harpaz, Iris; Seliktar, Dror

    2013-08-01

    Mesenchymal stromal cells residing in proteolytically responsive hydrogel scaffolds were subjected to changes in mechanical properties associated with their own three-dimensional (3-D) morphogenesis. In order to investigate this relationship the current study documents the transient degradation and restructuring of fibroblasts seeded in hydrogel scaffolds undergoing active cell-mediated reorganization over 7days in culture. A semi-synthetic proteolytically degradable polyethylene glycol-fibrinogen (PF) hydrogel matrix and neonatal human dermal fibroblasts (NHDF) were used. Rheology (in situ and ex situ) measured stiffening of the gels and confocal laser scanning microscopy (CLSM) measured cell morphogenesis within the gels. The assumption that the matrix modulus systematically decreases as cells locally begin to enzymatically disassemble the PF hydrogel to become spindled in the material was not supported by the bulk mechanical property measurements. Instead, the PF hydrogels exhibited cell-mediated stiffening concurrent with their dynamic morphogenesis, as indicated by a four-fold increase in storage modulus after 1week in culture. Fibrin hydrogels, which were used as the control biomaterial, proved similarly adaptive to cell-mediated remodeling only in the presence of the exogenous serine protease inhibitor aprotinin. Acellular and non-viable hydrogels also served as control groups to verify that transient matrix remodeling was entirely associated with cell-mediated events, including collagen deposition, cell-mediated proteolysis, and the formation of multicellular networks within the hydrogel constructs. The fact that cell network formation and collagen deposition both paralleled transient stiffening of the PF hydrogels, further reinforces the notion that cells actively balance between proteolysis and ECM synthesis when remodeling proteolytically responsive hydrogel scaffolds. PMID:23624218

  3. Human influenza A viruses are proteolytically activated and do not induce apoptosis in CACO-2 cells

    International Nuclear Information System (INIS)

    Replication of human influenza A/H3N2 and A/H1N1 viruses was studied in human CACO-2 cells, a continuous line of intestinal epithelial differentiated cells. Hemagglutinin (HA) was cleaved in these cells by an endogenous protease. Thus, infectious virus was produced that underwent multiple cycle replication and plaque formation in the absence of trypsin added to the media. Cleavage of de novo-synthesized HA occurred at a late stage of the exocytic pathway as indicated by pulse-chase labeling and by experiments employing endoglycosidase H and brefeldin A treatment. However, surface-labeling experiments employing biotinylation suggested that there is no cleavage at the plasma membrane. Unlike HA of serotypes H5 and H7 cleaved at multibasic cleavage sites by furin, the HAs with monobasic cleavage sites analyzed here were not cleaved in CACO-2 cells in the presence of aprotinin, a natural inhibitor of trypsinlike proteases. Growing CACO-2 cells were able to cleave HA of incoming virus, although influenza virus activating protease was not detected in culture medium. These observations indicate that the activating enzyme of CACO-2 cells is a trypsinlike protease functioning in the trans-Golgi network and presumably endosomes. In support of this concept immune staining with antibodies specific to human and bovine trypsin revealed the presence of a trypsinlike protease in CACO-2 cells. Unlike MDCK and CV-1 cells undergoing rapid apoptosis after influenza virus infection, CACO-2 cells showed no apoptosis but displayed cytopathic effects with necrotic signs significantly later after infection. It follows from these data that, depending on the cell type, influenza virus may kill cells either by apoptosis or by necrosis

  4. Leptospira interrogans Lsa23 protein recruits plasminogen, factor H and C4BP from normal human serum and mediates C3b and C4b degradation.

    Science.gov (United States)

    Siqueira, Gabriela H; Atzingen, Marina V; de Souza, Gisele O; Vasconcellos, Silvio A; Nascimento, Ana L T O

    2016-02-01

    It has been reported that pathogenic Leptospira are resistant to normal human serum (NHS) due to their ability to evade the complement immune system by interacting with factor H (FH) and C4b-binding protein (C4BP) regulators. Moreover, plasmin generation on the leptospiral surface diminishes C3b and IgG deposition, decreasing opsonophagocytosis by immune competent cells. We have previously reported that Lsa23 (LIC11360) is a multipurpose protein capable of binding purified extracellular matrix molecules, FH, C4BP and plasminogen (PLG)/plasmin in the presence of PLG activators. In this work, we provide further evidence that Lsa23 is located at the bacterial surface by using immunofluorescence microscopy. We show that Lsa23 has the ability to acquire FH, C4BP and PLG from NHS, and use these interactions to evade innate immunity. The binding with the complement regulators FH and C4BP preserves factor I (FI) activity, leading to C3b and C4b degradation products, respectively. C3b and C4b alpha-chain cleavage was also observed when Lsa23 bound to PLG generating plasmin, an effect blocked by the protease inhibitor aprotinin. Lsa23 also inhibited lytic activity by NHS mediated by both classical and alternative complement pathways. Thus, Lsa23 has the ability to block both pathways of the complement system, and may help pathogenic Leptospira to escape complement-mediated clearance in human hosts. Indeed, NHS treated with Lsa23 confers a partial serum resistance phenotype to Leptospira biflexa, whereas blocking this protein with anti-Lsa23 renders pathogenic L. interrogans more susceptible to complement-mediated killing. Thus, Lsa23 is a multifunctional protein involved in many pathways, featuring C4b cleavage by plasmin, knowledge that may help in the development of preventive approaches to intervene with human complement escape by this versatile pathogen. PMID:26614523

  5. 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. PMID:26183462

  6. Extracellular matrix is a source of mitogenically active platelet-derived growth factor.

    Science.gov (United States)

    Field, S L; Khachigian, L M; Sleigh, M J; Yang, G; Vandermark, S E; Hogg, P J; Chesterman, C N

    1996-08-01

    Platelet-derived growth factor (PDGF) is a chemotactic and mitogenic agent for fibroblasts and smooth muscle cells and plays a key role in the development of atherosclerotic lesions. PDGF is produced by a number of normal and transformed cell types and occurs as homo- or heterodimers of A and B polypeptide chains. Using Chinese hamster ovary (CHO) cells transfected with various forms of PDGF, we have previously shown that PDGF A(s) (short splice version) is secreted, PDGF A(l) (long splice version) predominantly extracellular matrix-associated, and PDGF B divided between medium, cells, and matrix. In the present study we have demonstrated the mitogenic activity of matrix-localized PDGF in artificial and more physiologically relevant models by culturing Balb/c-3T3 cells (3T3), human foreskin fibroblasts (HFF), and rabbit aortic smooth muscle cells (SMC) on extracellular matrix (ECM) laid down by PDGF-expressing CHO cells and human umbilical vein endothelial cells (HUVEC). These cells responded to the local growth stimulus of PDGF-containing CHO ECM and HUVEC ECM. We showed that 3T3 cells required proteolytic activity to utilize matrix-localized PDGF, as aprotinin and epsilon-ACA inhibited growth and 3T3 cells were shown to possess plasminogen activator activity. HFF and SMC did not appear to require proteolytic activity (including metalloproteinase and serine protease activity) as a prerequisite for mitogenesis but were able to access immobilized PDGF by contact with the matrix. An understanding of the mechanisms whereby the utilization of stored PDGF is controlled in situations of excessive cellular proliferation will aid in the development of therapy for these conditions. PMID:8707868

  7. L-lysine escinat, thiotriazolin, gordox and mydocalm influence on oxygen tension in the intestinal wall and acid-base balance and limited proteolysis in intestinal venous blood in terms of intraabdominal hypertension modeling

    Directory of Open Access Journals (Sweden)

    Sapegin V.I.

    2014-11-01

    Full Text Available In acute experiments on rabbits there were studied changes in oxygen tension in the intestinal wall tissues, acid-base balance and limited proteolysis and its inhibitors in intestinal venous blood, protective action of L-lysine escinat (0,15 mg/kg / single dose, thiotriazolin (25 mg/kg / single dose, aprotinin (gordox (10,000 units/kg / single dose in sequential modeling of standard levels increasing of intra-abdominal hypertension (IAH — 50, 100, 150, 200, 250, 300, 350 m H2O, and also of tolperison (mydocalm (5 mg/kg / single dose on modeling of stable 3-hour IAH 200 m H2O. The IAH modeling was performed by means of stand of our construction. Under the influence of IAH the compensated metabolic acidosis in intestinal venous blood with a compensative hyperpnoe develops, decline of oxygen tension in tissues and activating of a limited proteolysis as well as decline of its inhibitors activity in intestinal venous blood occur. By the degree of metabolic acidosis prevention investigational preparations were distributed as follows gordox > thiotriazolin = L-lysine escinat = mydocalm, and by prevention of decline of oxygen tension in tissues — thiotriazolin > L-lysine escinat > mydocalm > gordox, it is is connected with different rate of methabolic products excretion into the blood, due to the influence on blood circulation and transcapilary exchange. By the degree of prevention of proteolytic activity and inhibitory potential changes, investigational preparations were distributed as follows: gordox > mydocalm > thiotriazolin > L-lysine escinat, this is connected with inhibition of proteolysis in gordox, and in other ones – with reduction of ischemic damage of tissues. Owing to different mechanism of action thiotriazolin, L-lysine escinat and mydocalm may be simultaneously recommended for a conservative treatment of patients with intraabdominal hypertension syndrome.

  8. 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.

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

    International Nuclear Information System (INIS)

    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

  10. 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.

  11. Novel approaches to tumor imaging in mice: radiolabeled epidermal growth factor

    International Nuclear Information System (INIS)

    Full text. We had earlier shown that the epidermal growth factor peptide (EGF) may be radio labelled with 99 m Tc at room temperature and neutral pH by using the NHS ester of S-acetyl MAG3 as f bifunctional chelator. By a competition binding assay, we found that MAG3-conjugated EGF retained biological activity. Furthermore, the labelled peptide exhibited saturation binding to EGF receptor-positive tumor cells lines which could be inhibited by presaturation of the cells with unlabeled, native EGF. Biodistribution in normal mice at 3 h post administration showed rapid clearance with minimal retention of the label in sampled organs. We have now investigated the tumor localization properties in mice of this labeled peptide. Nude mice implanted with the EGF receptor-positive tumors A431 and LS-174T were administered labeled EGF and a labeled control peptide (BPTI, aprotinin. Tumour uptake at 12 h post IV administration was 0.44% ID/gm for EGF vs. 0.09%ID/gm for the control. Pretreatment of tumored mice with unlabeled EGF blocked about half the tumor uptake. Animals were also administered an anti-EGF receptor antibody labeled with 99 mTc via MAG3. Relative to the antibody, tumor to muscle ratios were improved from 6 to 15 and tumor to muscle ratios were improved from 6 to 15 and tumor to blood ratios from 0.4 to 7 with EGF. These favorable results along with documented evidence of overexpression of the EGF receptor in many human tumors suggest that 99 mTc-EGF should be further considered for tumor detection

  12. Temporal and pharmacological characterization of angiostatin release and generation by human platelets: implications for endothelial cell migration.

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    Aneta Radziwon-Balicka

    Full Text Available Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis. Amongst these factors is the angiogenesis inhibitor angiostatin, which is released during thrombus formation. The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown. Hence, our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration, an early stage of angiogenesis. We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets. Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor (VEGF. Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin. Acetylsalicylic acid, MRS2395, GPIIb/IIIa blocking peptide, and aprotinin were used to characterize platelet angiostatin release and generation. An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition. Compared to VEGF, angiostatin generation and release from α-granules occurred later temporally during platelet aggregation. Consequently, collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets. Platelet inhibitors prostacyclin, S-nitroso-glutathione, acetylsalicylic acid, and GPIIb/IIIa blocking peptide, but not a P2Y12 inhibitor, suppressed angiostatin release but not generation. Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration. Hence, the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo

  13. 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

  14. 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.

  15. Some physical and chemical properties of the smooth muscle inhibitory factor in extracts of the bovine retractor penis muscle.

    Science.gov (United States)

    Gillespie, J S; Hunter, J C; Martin, W

    1981-06-01

    1. A method of extracting and partially purifying a smooth muscle inhibitory factor from the bovine retractor penis is described. This consists of extraction in methanol followed by adsorption on an anion exchange resin, elution from the resin with 500 mM-sodium chloride solution and, if necessary, removal of adenine nucleotides by adsorption on alumina. 2. The inhibitory factor exists in a stable pharmacologically inactive form and an unstable pharmacologically active form. Conversion to the active form is by a brief exposure to acid at pH 2.0. 3. The inhibitory factor is insoluble in ether or acetone but soluble in methanol. Anhydrous methanol, however, irreversibly destroys pharmacological activity especially if the inhibitory factor is in the active form. This effect of methanol is prevented by the presence of 20-30-% water. 4. The inhibitory factor binds to an anion exchange resin but not to a cation exchange resin. It can be eluted from the resin by 500 mM-sodium chloride solution. 5. The molecular weight of the inhibitory factor, as judged by the ability to pass ultrafiltration membranes, is about 500. 6. Inhibitory activity is unaffected by the proteases trypsin, subtilisin or pepsin or by leucine aminopeptidase, pyroglutamate aminopeptidase or carboxypeptidase. The inhibitory effect of the extract and the inhibitory response to stimulation of the non-adrenergic, non-cholinergic nerves are also unaffected by the protease inhibitor, aprotinin. The active material, therefore, is unlikely to be a peptide. 7. Inhibitory activity is abolished by exposure of the extracts to periodic acid or sodium periodate. Acetic anhydride in pyridine also abolishes activity but the vehicle pyridine is also effective. 8. Sodium borohydride but not borate abolishes inhibitory activity when added to the acid-activated material at pH 2.0 but has no effect or may even potentiate activity if added to the stable inactive form at pH 9.0. When added to the acid-activated but

  16. Weight-based dosing in medication use: what should we know?

    Science.gov (United States)

    Pan, Sheng-dong; Zhu, Ling-ling; Chen, Meng; Xia, Ping; Zhou, Quan

    2016-01-01

    Background Weight-based dosing strategy is still challenging due to poor awareness and adherence. It is necessary to let clinicians know of the latest developments in this respect and the correct circumstances in which weight-based dosing is of clinical relevance. Methods A literature search was conducted using PubMed. Results Clinical indications, physiological factors, and types of medication may determine the applicability of weight-based dosing. In some cases, the weight effect may be minimal or the proper dosage can only be determined when weight is combined with other factors. Medications within similar therapeutic or structural class (eg, anticoagulants, antitumor necrosis factor medications, P2Y12-receptor antagonists, and anti-epidermal growth factor receptor antibodies) may exhibit differences in requirements on weight-based dosing. In some cases, weight-based dosing is superior to currently recommended fixed-dose regimen in adult patients (eg, hydrocortisone, vancomycin, linezolid, and aprotinin). On the contrary, fixed dosing is noninferior to or even better than currently recommended weight-based regimen in adult patients in some cases (eg, cyclosporine microemulsion, recombinant activated Factor VII, and epoetin α). Ideal body-weight-based dosing may be superior to the currently recommended total body-weight-based regimen (eg, atracurium and rocuronium). For dosing in pediatrics, whether weight-based dosing is better than body surface-area-based dosing is dependent on the particular medication (eg, methotrexate, prednisone, prednisolone, zidovudine, didanosine, growth hormone, and 13-cis-retinoic acid). Age-based dosing strategy is better than weight-based dosing in some cases (eg, intravenous busulfan and dalteparin). Dosing guided by pharmacogenetic testing did not show pharmacoeconomic advantage over weight-adjusted dosing of 6-mercaptopurine. The common viewpoint (ie, pediatric patients should be dosed on the basis of body weight) is not always

  17. 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

  18. In vitro comparison of renal handling and uptake of two somatostatin receptor-specific peptides labeled with indium-111

    International Nuclear Information System (INIS)

    Radiolabeled receptor-specific somatostatin analogs labeled with gamma- or beta-emitting radionuclides are useful for scintigraphic imaging and/or therapy of selected neuroendocrine tumors. However, significant renal uptake may result in radiotoxicological injury of the kidney and can limit clinical application of the agents. The aim of the study was to analyze renal handling, rate, and mechanism of renal accumulation of two somatostatin receptor-targeted peptides, [DOTA0, Tyr3, Thr8]-octreotide (DOTA-TATE) and [DOTA0, 1-Nal3]-octreotide (DOTA-NOC), labeled with indium-111 using in vitro methods. The perfused rat kidney and freshly isolated rat renal cells were used as experimental models. The perfusion was performed in a recirculation regimen at constant pressure with solution containing bovine albumin, erythrocytes, and a mixture of essential substrates. The renal cells were isolated from rat kidneys using two-phase collagenase perfusion. Accumulation studies were used to evaluate the renal uptake of the peptides and to compare their accumulation with that of passively or actively transported model drugs. The influence of selected inhibitors of receptor-mediated endocytosis and the inhibition of energy-dependent transport processes on the uptake were also investigated using isolated renal cells. The renal clearance of 111In-DOTA-NOC in the perfused rat kidney was significantly lower than that of 111In-DOTA-TATE. Reverse situation was found in the case of renal retention. Pretreatment of the perfused kidney with maleate markedly decreased the renal retention. 111In-DOTA-NOC was accumulated in the isolated renal cells at a higher rate than 111In-DOTA-TATE (ratio 3:1). The uptake of the radiopeptides in renal cells was higher than the uptake of not only the passively transported sucrose but also actively transported and accumulated methylglucose. The rank order of potency to inhibit the uptake by active endocytosis was approximately aprotinin>maleate>lysine. The

  19. 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.

  20. 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

  1. Weight-based dosing in medication use: what should we know?

    Directory of Open Access Journals (Sweden)

    Pan SD

    2016-04-01

    Full Text Available Sheng-dong Pan,1 Ling-ling Zhu,2 Meng Chen,3 Ping Xia,1 Quan Zhou3 1Division of Medical Administration, 2VIP Care Ward, Division of Nursing, 3Department of Pharmacy, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of China Background: Weight-based dosing strategy is still challenging due to poor awareness and adherence. It is necessary to let clinicians know of the latest developments in this respect and the correct circumstances in which weight-based dosing is of clinical relevance. Methods: A literature search was conducted using PubMed. Results: Clinical indications, physiological factors, and types of medication may determine the applicability of weight-based dosing. In some cases, the weight effect may be minimal or the proper dosage can only be determined when weight is combined with other factors. Medications within similar therapeutic or structural class (eg, anticoagulants, antitumor necrosis factor medications, P2Y12-receptor antagonists, and anti-epidermal growth factor receptor antibodies may exhibit differences in requirements on weight-based dosing. In some cases, weight-based dosing is superior to currently recommended fixed-dose regimen in adult patients (eg, hydrocortisone, vancomycin, linezolid, and aprotinin. On the contrary, fixed dosing is noninferior to or even better than currently recommended weight-based regimen in adult patients in some cases (eg, cyclosporine microemulsion, recombinant activated Factor VII, and epoetin α. Ideal body-weight-based dosing may be superior to the currently recommended total body-weight-based regimen (eg, atracurium and rocuronium. For dosing in pediatrics, whether weight-based dosing is better than body surface-area-based dosing is dependent on the particular medication (eg, methotrexate, prednisone, prednisolone, zidovudine, didanosine, growth hormone, and 13-cis-retinoic acid. Age-based dosing strategy is better than weight

  2. 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

    Directory of Open Access Journals (Sweden)

    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

  3. 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

  4. 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

  5. Monograph: Concluding Remarks

    Directory of Open Access Journals (Sweden)

    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