Fibrinolysis and Proteolysis最新文献

{"title":"Fibrin(ogen) degradation by a 24k-endopeptidase from a Chryseobacterium Sp.","authors":"H. Lijnen, B. Hoef, I. Roelants, D. Collen","doi":"10.1054/FIPR.2000.0076","DOIUrl":"https://doi.org/10.1054/FIPR.2000.0076","url":null,"abstract":"Abstract Summary A novel 24k-endopeptidase purified from the conditioned medium of a novel species of the Chryseobacterium genus, degraded purified fibrinogen and fibrin. At an enzyme/substrate ratio of 1/200 to 1/1000 rapid degradation of the human fibrinogen α-chain occurred and delayed degradation of the β-chain, whereas the γ-chain was more resistant. Degradation was associated with loss of thrombin-inducible coagulation. Incubation of 24k-endopeptidase (500 nM) in human plasma at 37°C for 3 h resulted in a decrease in fibrinogen to 38 ± 7% of baseline, and an increase of the thrombin time from 21 ± 1 to 52 ± 17 s. Other coagulation factors as well as plasminogen were not significantly affected. Fibrinogen digested with 24k-endopeptidase did not confer anticoagulant properties when added to human plasma. In a buffer milieu,125I-fibrin labelled matrix was lysed in a time- and concentration-dependent manner by 24k-endopeptidase. Purified125I-labelled fibrin clots (65 μl) in a buffer milieu (500 μl) were also efficiently lysed, requiring 50 nM 24k-endopeptidase for 50% lysis in 1 h. The presence of a physiological fibrinogen concentration in this assay did not affect lysis. In contrast, no significant lysis of125I-fibrin labelled plasma clots (65 μl) in human plasma (500 μl) was observed upon incubation with 24k-endopeptidase (up to 1 μM) for up to 24 h. Incubation of 24k-endopeptidase with α2-macroglobulin resulted in formation of a complex with impaired proteolytic activity against fibrinogen. These findings indicate that 24k-endopeptidase may reduce plasma coagulability by degradation of fibrinogen and that its differential action on fibrin in a buffer milieu or in plasma may be due to inhibition by α2-macroglobulin.","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"59 1","pages":"247-252"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83962315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Does albumin play a role in fibrinolysis by its inhibition of plasminogen activation","authors":"M. V. D. Velden, H. Smolders, H. V. Rijn, H. Voorbij","doi":"10.1054/FIPR.2000.0067","DOIUrl":"https://doi.org/10.1054/FIPR.2000.0067","url":null,"abstract":"Abstract One of the clinical characteristics of patients with nephrotic syndrome is hypoalbuminemia. The mechanism underlying the susceptibility to thrombosis in this group of patients is not fully understood. The hypoalbuminemia may alter the structure of fibrin clots and affect clot lysis, thereby influencing the coagulolytic balance. In the present study, we investigated the effect of albumin on fibrinolysis. The effect of albumins from several manufacturers and recombinant albumin on the plasminogen activation were tested in an in vitro assay. A chromogenic substrate highly selective for plasmin was used to detect the generation of plasmin. This study revealed a dose-dependent inhibition of plasminogen activation by albumin in a concentration range between 12.5 g/L and 50 g/L. The percentage of inhibition at maximum plasmin activity was different for each albumin, varying from 23% to 90%. Pre-incubation of albumin with fibrinogen enhanced the inhibition, suggesting that by binding to fibrin(ogen) albumin may hinder the formation of the ternary complex and thereby interfere in the fibrinolytic process. This may have consequences for patients with nephrotic syndrome, whose low albumin levels may provide a mechanism partially compensating the well-known hypercoagulability.","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"25 1","pages":"242-246"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89675093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Upregulation of MMP-2 by all-trans retinoic acid is mediated by TGF- β 1 in cultured rat mesangial cell","authors":"W. Lin ,&nbsp;N. Zhang ,&nbsp;S. Zhang ,&nbsp;J. Gu ,&nbsp;M. Guo","doi":"10.1054/fipr.2000.0066","DOIUrl":"https://doi.org/10.1054/fipr.2000.0066","url":null,"abstract":"<div><p>Matrix metalloproteinase-2 (MMP-2) degrades basement membrane collagen and its abnormal expression is associated with glomerulonephritis and glomerulosclerosis. All-trans retinoic acid (ATRA) has been indicated as preventing age-related glomerulosclerosis. The results of our study showed that ATRA upregulated expression of MMP-2 mRNA and secretion of MMP-2 proteins, and increased enzymatic activity of MMP-2 in cultured mesangial cell. In addition, ATRA also caused a dose-dependent increase in the secretion of transforming growth factor-β (TGF-β1) peptides. Since TGF-β1 regulated the expression of MMP-2 in mesangial cell, it is possible that TGF-β1 is involved in ATRA-induced MMP-2 expression. Using antisense TGF-β1 RNA strategy, antisense TGF-β1 construct almost completely blocked secretion of active TGF-β1 peptides in the antisense-bearing transfectants. Northern blot analysis showed that the transfectants treated with 10^–6M ATRA for 72 h neither increased nor decreased the levels of MMP-2 messenger ribonucleic acid (mRNA). Moreover, addition of anti-TGF-β1 antibodies to mesangial cell in the presence of 10^–6M ATRA reduced ATRA-induced MMP-2 expression dose dependently. These data suggest that TGF-β1 might be involved in ATRA-induced MMP-2 expression.</p></div>","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"14 4","pages":"Pages 235-241"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1054/fipr.2000.0066","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71826463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Laboratory Techniques in Thrombosis – A Manual, 2nd Revised Edition of ECAT Assay Procedures","authors":"Professor M.M. Samama","doi":"10.1054/fipr.2000.0086","DOIUrl":"https://doi.org/10.1054/fipr.2000.0086","url":null,"abstract":"","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"14 4","pages":"Page 268"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1054/fipr.2000.0086","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71826465","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An increased thrombin generation is detectable for at least 1 week following elective percutaneous transluminal coronary angioplasty","authors":"D. Prisco, E. Antonucci, M. Capanni, L. Chiarugi, V. Boddi, C. Giglioli, M. Comeglio, S. Fedi, G. Gensini, R. Abbate","doi":"10.1054/FIPR.2000.0075","DOIUrl":"https://doi.org/10.1054/FIPR.2000.0075","url":null,"abstract":"Objective: The present prospective study was planned to investigate: (1) how long the early haemostatic changes after PTCA last and (2) if some coagulation and/or fibrinolytic parameters assessed during the first month after PTCA may be predictive of subsequent clinical recurrence. \u0000 \u0000Setting: Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence, Italy. \u0000 \u0000Material and Methods: In 72 patients undergoing PTCA fibrinogen, F1+2, TAT, D-dimer and ELT were evaluated before the procedure (T1) and 2 (T2), 7 (T7) and 30 days (T30) after PTCA; PAI-1 and t-PA were assessed before PTCA and after 7 and 30 days. Follow-up angiography was performed only in patients with recurrence of ischaemia or positive ergometric tests. \u0000 \u0000Results: F1+2, TAT and fibrinogen were significantly increased at T2 (P<0.005); after a week, F1+2 and fibrinogen were still significantly higher in comparison to baseline values (P<0.005). At T30 these parameters showed significantly lower levels if compared to T1 (P<0.005). Plasma D-dimer concentration significantly increased at T2 and T7 (P<0.001), but no difference was found between baseline values and those at T30. PAI-1 activity significantly decreased at T7 (P<0.001), whereas it was similar to baseline at T30. No significant variations of t-PA levels were observed at the different times. Finally, ELT significantly increased at T2 (P<0.001), but at T7 and T30 the values were similar to baseline values. Clinical recurrence occurred in 19 patients. The values of various parameters investigated were not different at any time considered between the patients with and without subsequent clinical recurrence. Heparin treatment had no significant influence on thrombin generation at different times whereas it had marginal influences on fibrinogen, PAI-1 and t-PA antigen levels. \u0000 \u0000Conclusion: A number of alterations in haemostasis takes place and persists 2 and 7 days after elective PTCA and heparin treatment is not able to blunt clotting activation. The haemostatic parameters assessed during the first month after PTCA seem not to be predictive of subsequent clinical recurrence.","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"40 1","pages":"253-260"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89683290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overexpression of plasminogen activator inhibitor type-1 in HT-1080 fibrosarcoma cells promotes lung colonization and in vitro cell adhesion","authors":"V.A Carroll,&nbsp;J Mihaly,&nbsp;Y Koshelnick,&nbsp;P Hufnagl,&nbsp;B.R Binder","doi":"10.1054/fipr.2000.0061","DOIUrl":"https://doi.org/10.1054/fipr.2000.0061","url":null,"abstract":"<div><p>Plasminogen activator inhibitor type 1 (PAI-1) is associated with tumour invasion, angiogenesis and metastatic spread. It is also a strong prognostic factor for relapse in a number of human cancers. This study was designed to investigate the effects of overexpression of PAI-1 on lung colonization by human HT-1080 fibrosarcoma cells. Full length PAI-1 cDNA was transfected in a non-aggressive HT-1080 clonal cell line (1–3C). Stable transfected clones were isolated of which one (3F52) secreted a 29-fold increase in PAI-1 protein levels (29.1±6.5 μg/10⁶cells/24 h) as compared with control mock transfected cells (1.0±0.2 μg/10⁶cells/24 h). 3F52 cells were significantly better able to form lung colonies after i.v. tail vein injection of athymic mice (mean number of colonies±SE 238±105) as compared with control cells (8±7). In addition, PAI-1 overexpressing cells were between 1.5 and 2.5 fold more adhesive to extracellular matrix membrane (ECM) proteins than mock transfected cells in an in vitro cell adhesion assay. These findings provide experimental support that PAI-1 facilitates tumour cell lodgement in vivo and adhesion of HT-1080 cells in vitro.</p></div>","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"14 4","pages":"Pages 215-220"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1054/fipr.2000.0061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71825985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Controlled evaluation of fibrinolytic response to intraoperative pneumatic intermittent sequential compression of the lower extremities during laparoscopic cholecystectomy","authors":"W. Schwenk, S. Ziemer, J. Neudecker, T. Kuntz, Joachim M. Müller","doi":"10.1054/FIPR.2000.0058","DOIUrl":"https://doi.org/10.1054/FIPR.2000.0058","url":null,"abstract":"Abstract Objective: During a pneumoperitoneum intraoperative intermittent sequential pneumatic compression (ISC) neutralizes venous stasis in the lower extremities and may also have an influence on systemic fibrinolytic activity. Design: Controlled study to evaluate whether ISC stimulates intravascular fibrinolytic activity. Setting: University-Hospital. Materials: 26 patients underwent laparoscopic cholecystectomy with (+ISC, n = 10) or without (–ISC, n = 16) ISC. Intervention: Intraoperative intermittent sequential compression of the lower extremities. Main outcome measures: Major endpoint – postoperative plasma activity of tissue-plasminogen-activator (t-PA). Minor endpoints – postoperative plasminogen-activator-inhibitor-1 (PAl-1) activity, plasmin-antiplasmin-complex (PAP) concentration, concentration of total fibrin degradation products (TDP), and D-Dimer concentration. Results: There was no difference in age, sex, body mass index (BMI), and properative fibrinolytic activity between the groups. Postoperative t-PA-activity (P =0.3), PAI-1-activity (P =0.9), PAP- (P =0.5), TDP-(P =0.2), and D-Dimer-concentrations (P =0.1) were also not different between both groups. Conclusion: During laparoscopic cholecystectomy intraoperative ISC does not activate systemic intravascular fibrinolysis. Because of the positive hemodynamic effect of ISC during pneumoperitoneum, studies of the antithrombotic efficacy of this tool are needed in the setting of laparoscopic surgery.","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"6 1","pages":"229-234"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78833311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Apoptosis","authors":"S. Nagata","doi":"10.1054/fipr.2000.0074","DOIUrl":"https://doi.org/10.1054/fipr.2000.0074","url":null,"abstract":"<div><p>The term ‘apoptosis’ was coined more than 30 years ago to define a cell death process that is accompanied by condensation of cytoplasm, loss of plasma membrane microvilli, and segmentation of the nucleus. This process is characterized biochemically by the extensive degradation of chromosomal DNA into oligomers of 180 bp. The underlying molecular mechanisms of apoptosis, as well as its physiological and pathological roles, were elusive for a long time. Recent studies have rapidly identified molecules that regulate the apoptotic process, and the physiological and pathological roles of this process are emerging. Here, I summarize the current knowledge on apoptotic cell death.</p></div>","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"14 2","pages":"Pages 82-86"},"PeriodicalIF":0.0,"publicationDate":"2000-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1054/fipr.2000.0074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71826232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulation of fibrinolytic activity by localization of inhibitors to fibrin(ogen)","authors":"N.A. Booth","doi":"10.1054/fipr.2000.0071","DOIUrl":"https://doi.org/10.1054/fipr.2000.0071","url":null,"abstract":"<div><p>Fibrin plays a key role in fibrinolysis, acting not only as a substrate but also as a regulator of activity. As well as stimulating plasminogen activation, it controls interactions between proteases and inhibitors, both protecting proteases from inhibition and, conversely, localizing inhibitors. The principal inhibitor of plasmin, α₂-antiplasmin, is cross-linked to fibrinogen and fibrin, inhibiting fibrinolysis. Our studies have shown that a second inhibitor, PAI-2, is also cross-linked to fibrinogen and fibrin, by either factor XIIIa or tissue transglutaminase. These inhibitors are both members of the serpin family but the cross-linking sites are quite unrelated. Cross-links are formed between glutamine residues in the inhibitors and lysine residues in fibrin(ogen). The Gln residues involved are at position 2 in the N-terminus of α₂-AP and at position 83 and 86 in PAI-2, located in a loop between helices C and D. All cross-linking observed was to the Aα chain of fibrin(ogen). The two inhibitors did not compete for cross-linking sites. α₂-AP binds only to Lys 303 of the Aα chain and a 30-residue peptide based on the sequence around this Lys competed with fibrinogen for cross-linking to α₂-AP but not for cross-linking to PAI-2. PAI-2 was cross-linked to several Lys residues (but not Lys 303) in the Aα chain, as shown by tryptic digestion and mass spectrometry. PAI-2 was cross-linked to Lys 148, 176, 183 and 467 by tissue transglutaminase and to Lys 148, 176, 230 and 413 by factor XIIIa. The activity of PAI-2 was not affected by cross-linking, so that this is a mechanism whereby it can be covalently bound to fibrinogen and retained in a fibrin clot, without loss of activity towards u-PA and two-chain t-PA. PAI-1, the other major inhibitor of fibrinolysis, also binds to fibrin but we find no evidence for its being cross-linked. All three inhibitors achieve high local concentrations on fibrin, which they protect from lysis by t-PA, u-PA and plasmin. The inhibitors differ in their major sources in blood, with α₂-AP present at high concentrations in plasma, PAI-1 primarily in platelets, and PAI-2 a product of stimulated monocytes, giving them distinct and complementary roles in stabilizing fibrin in different physiological and pathological locations.</p></div>","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"14 2","pages":"Pages 206-213"},"PeriodicalIF":0.0,"publicationDate":"2000-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1054/fipr.2000.0071","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71826157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fibrinogen functions and fibrin assembly","authors":"M.W. Mosesson","doi":"10.1054/fipr.2000.0054","DOIUrl":"https://doi.org/10.1054/fipr.2000.0054","url":null,"abstract":"<div><p>Fibrinogen and fibrin play important roles in clot formation, fibrinolysis, cellular and matrix interactions, inflammation, and wound healing. These biological events are regulated to a large extent by clot formation itself and by complementary interactions between specific reactive sites on fibrin(ogen) and extrinsic molecules such as enzymes, proteins including other clotting factors, or cell receptors. Fibrinogen is comprised of two sets of three polypeptide chains termed Aα, Bβ, and γ, all six of which are joined by disulfide bridges to form the amino-terminal E domain. The molecules are elongated trinodular structures consisting of two globular outer D domains that are connected to its central E domain by a coiled-coil segment. These domains contain constitutive binding sites (e.g. Da, Db, γXL, D:D, γ′, thrombin substrate, platelet receptor, leukocyte integrin receptor) as well as interactive sites that become expressed as a result of fibrinogen cleavage by thrombin or that are exposed as a consequence of polymerization (e.g. t-PA binding sites). Other relevant constitutive sites in fibrinogen include two thrombin substrate recognition sites in each E domain plus a high-affinity non-substrate thrombin binding site in each γ′ chain that that also binds factor XIII. Constitutive binding sites on fibrinogen participate in fibrin assembly by self-association (γXL or D:D) or by complementary association with exposed fibrin sites (Da to EA and Db to E <span><math><mtext>B</mtext></math></span>). Other relevant constitutive sites in fibrin include a low-affinity thrombin-binding site in the fibrin E domain that evidently remains as a residual of the substrate binding site.</p><p>Fibrin polymerization is initiated by thrombin cleavage of fibrinopeptide A (FPA) from fibrinogen Aα chains, exposing two E domain E <span><math><mtext>A</mtext></math></span> sites. Cleavage of fibrinopeptide B (FPB) from Bβ chains exposes another E domain polymerization site, E <span><math><mtext>B</mtext></math></span>, that also interacts with platelets, fibroblasts and endothelial cells. Fibrin generation is followed by end-to-middle intermolecular D-to-E associations forming linear and equilaterally branched double-stranded fibrils, and is accompanied by lateral fibril associations that form multi-stranded fibers. Concomitantly, thrombin-activated factor XIIIa introduces covalent crosslinks into these polymers, mainly between γ chains (at γXL sites forming γ-dimers) and α chains (α-polymers), to complete the mature clot network structure.</p></div>","PeriodicalId":100526,"journal":{"name":"Fibrinolysis and Proteolysis","volume":"14 2","pages":"Pages 182-186"},"PeriodicalIF":0.0,"publicationDate":"2000-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1054/fipr.2000.0054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71826243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}