MH_dev_154

Growth associated proteins in tumor cells and stroma related to disease progression of colon cancer accounting for tumor tissue DB00917 content . Connections among specific proteins ( Bax , Bcl - 2 , P09038 REA , P23219 REA , P35354 REA , E-cad , p15 , p53 , P12004 REA , TGFbeta 3 , TUNEL , P04275 REA ) in control of cell proliferation , apoptosis , cell adhesion , tumor vascularity and DB00917 content were evaluated in colon cancer as related to disease progression and survival . Tumor tissue and adjacent normal colon mucosa were obtained at curative resection in 22 patients . DB00917 concentrations were assessed in tumor tissue and tumor derived blood , splanchnic blood , peripheral venous blood and urine . Host inflammation was determined ( CRP , P03372 REA ) in relationship to tumor differentiation and stage . Patients survived as expected according to Dukes A-D staging . Growth-related proteins correlated between tumor cells and stroma as well as between protein factors within tumor cells and tumor stroma . P35354 REA predicted tumor tissue content of DB00917 ( p < 0.002 ) , without reflection in tumor derived blood . Systemic inflammation was predicted by p15 , TGFbeta 3 and Bcl - 2 in tumor tissue ( p < 0.001 ) . p15 and P04275 REA predicted reduced survival in ungrouped patients ( p < 0.02 ) , while p15 , P12004 REA , TGFbeta 3 and P04275 REA predicted reduced survival ( p < 0.0001 ) when patient grouping accounted for high tumor content of DB00917 . Our results connect systemic inflammation and survival to P35354 REA staining and increased DB00917 in colon cancer . Thus , it seems important to understand proximal signals behind upregulation of P35354 REA and subsequent DB00917 production in certain tumor cells in colon cancer .

DB00013 SUB plasminogen activator , Q03405 REA , P08253 REA , and P14780 REA in the P13671 REA - glioblastoma rat model . BACKGROUND : In glioblastoma multiforme ( GBM ) , the serine protease urokinase plasminogen activator ( uPA ) , and matrix metalloproteases ( P08253 REA / P14780 REA ) contribute to its invasive growth pattern , which is the major obstacle to successful surgical treatment . MATERIALS AND METHODS : The expression of uPA was determined in monolayers and spheroids of the rodent GBM cell line P13671 REA by immunohistochemistry and polymerase chain reaction ( PCR ) . The longitudinal expression of proteases was studied in orthotopically implanted spheroids by semi-quantitative immunohistochemistry ( IHC ) in Sprague Dawley rats ( n = 40 ) . The tumor volume was monitored by magnetic resonance imaging ( Q9BWK5 ) . RESULTS : In vitro , the GBM cell line P13671 REA expresses high levels of uPA . In vivo , a continuous increase of uPA , uPA-receptor ( Q03405 REA ) , P08253 REA , and P14780 REA expression was found in the infiltration zone . uPA was located exclusively in the infiltration zone and in the vascular basal layers . The mean tumor volume 23 days after implantation was 3.2 mm3 . CONCLUSION : uPA , Q03405 REA , P08253 REA and P14780 REA play an important role in GBM growth . Blockade of uPA and interruption of the proteolytic cascade could become a useful tool in the therapy of GBM .

A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 REA and P35354 REA ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 REA ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg .

Angiogenic alterations associated with circulating neoplastic DNA in ovarian carcinoma . OBJECTIVES : Forty percent of women with ovarian carcinoma have circulating free neoplastic DNA identified in plasma . Angiogenesis is critical in neoplastic growth and metastasis . We sought to determine whether circulating neoplastic DNA results from alterations in the balance of angiogenesis activators and inhibitors . METHODS : Sixty patients with invasive ovarian carcinomas with somatic P04637 REA mutations that had been characterized for circulating neoplastic DNA had carcinoma analyzed for microvessel density using immunohistochemistry with CD31 and for the expression of P15692 REA , Q15389 REA , O15123 REA , P35354 REA , P00749 REA , P07996 REA , P09603 REA , P42336 REA , Q16665 REA , P10145 REA , P08253 REA , and P14780 REA message by real-time quantitative polymerase chain reaction . The expression of each gene was calculated relative to P04406 REA expression for each neoplasm . Patient plasma had been tested for circulating neoplastic DNA using a ligase detection reaction . RESULTS : P08253 REA expression was significantly correlated with free plasma neoplastic DNA ( P = . 007 ) . Microvessel density was not correlated with plasma neoplastic DNA or P38398 REA / 2 mutation status . The expression pattern of other angiogenic factors did not correlate with plasma neoplastic DNA but correlated with each other . P38398 REA / 2 mutated carcinomas had significantly different expression profiles of angiogenesis activators and inhibitors in comparison to sporadic carcinomas . CONCLUSIONS : P08253 REA expression is associated with the presence of circulating neoplastic DNA in women with ovarian carcinoma . These data are consistent with the proinvasive properties of P08253 REA and suggest that the presence of circulating neoplastic DNA indicates a more aggressive malignant phenotype . Carcinomas with germ line P38398 REA / 2 mutations had a lower angiogenic profile than those without mutations .

Confocal fluorescence microscopy of urokinase plasminogen activator receptor and cathepsin D in human MDA-MB - 231 breast cancer cells migrating in reconstituted basement membrane . Using confocal fluorescence microscopy with a monoclonal antibody , we have localized the receptor for urokinase plasminogen activator ( Q03405 REA ) in MDA-MB - 231 human breast cancer cells migrating into a reconstituted basement membrane . Patchy and polarized Q03405 REA immunoreactivity was found at the cell membrane , and strong staining was found both in the ruffled border or leading edge of the cells and at pseudopodia penetrating into the membrane . Intracellular Q03405 REA staining was localized in the paranuclear region and in rounded granule-like structures ; some of these were identified as lysosomes by double staining for Q03405 REA and the lysosomal enzyme cathepsin D . DB00013 SUB plasminogen activator ( uPA ) activity has previously been shown to play a role in migration of cells into basement membranes , and it has been proposed that Q03405 REA also is involved in this process . uPA is known to be internalized and degraded after complex formation with the inhibitor P05121 REA . Lysosomal Q03405 REA immunoreactivity may result from concomitant internalization of the receptor .

Transforming growth factor-beta is a strong and fast acting positive regulator of the level of type - 1 plasminogen activator inhibitor mRNA in WI - 38 human lung fibroblasts . We have studied the mechanism of a transforming growth factor-beta ( TGF-beta ) - stimulated production of type - 1 plasminogen activator inhibitor ( P05121 REA ) in WI - 38 human lung fibroblasts . TGF-beta causes an early increase in the P05121 REA mRNA level which reaches a maximal 50 - fold enhancement after 8 h . Blocking of protein synthesis with cycloheximide causes an equally strong increase in the level of P05121 REA mRNA . Quantitative studies of the effect of TGF-beta on P05121 REA protein levels in cell extracts and culture media by using monoclonal antibodies are consistent with the effect on P05121 REA mRNA . The results suggest a primary effect of TGF-beta on P05121 REA gene transcription , and also suggest the possibility that the transcription of this gene in non-induced cells may be suppressed by a short-lived negatively regulating protein . DB00013 SUB - type ( u-PA ) and tissue-type ( t-PA ) plasminogen activators are decreased in the culture media of TGF-beta-treated cells concomitantly with the increase in P05121 REA accumulation . These findings show that a primary and important biological effect of TGF-beta may be an overall decreased extracellular proteolytic activity , and give an insight into the molecular mechanisms underlying TGF-beta action at the genetic level .

P12821 REA inhibition actively promotes cell survival by altering gene expression . We tested the effect of P12821 REA inhibition on the survival of bovine retinal ( Q9UIJ5 ) and choroidal ( CEC ) endothelial cells ( EC ) in culture . The P12821 REA inhibitor captopril delayed the apoptotic tube collapse of Q9UIJ5 on Matrigel for > 15 days . DB01197 MEN treatment of confluent monolayers ( 2-8 weeks ) followed by slow starvation ( 2-4 weeks ) increased EC viability by approximately 200 % . Two-week captopril exposures were sufficient to confer maximal protection . Only vehicle-treated EC demonstrated apoptotic features such as membrane blebbing and DNA laddering . By RT-PCR , the starvation marker p202 was upregulated only in starved cells . In Q9UIJ5 , captopril upregulated the pro-survival proteins mortalin - 2 , uPA , and Q03405 REA while downregulating the anti-growth sprouty - 4 and tPA . In CEC , captopril also upregulated tPA and its inhibitor P05121 REA . DB00594 ( uPA inhibitor ) blocked the captopril-induced increase in EC survival , secondary sprouting , and invasion in Matrigel . The pro-survival effects of captopril involve the reprogramming of genes involved in cell survival and immortalization .

Expression of active plasminogen activator inhibitor - 1 reduces cell migration and invasion in breast and gynecological cancer cells . DB00013 SUB - type ( uPA ) plasminogen activator is regulated by serine protease inhibitors ( serpins ) , especially plasminogen activator inhibitor - 1 ( P05121 REA ) . In many cancers , uPA and P05121 REA contribute to the invasive phenotype . We examined the in vitro migration and invasive capabilities of breast , ovarian , endometrial , and cervical cancer cell lines compared to their plasminogen activator system profiles . We then overexpressed active wild-type P05121 REA and an inactive " substrate " P14 form of P05121 REA ( T333R ) using stable transfection and adenoviral gene delivery . We also upregulated endogenous uPA and P05121 REA in these cells by treatment with transforming growth factor-beta . Some breast and ovarian cancer cell lines with natural expression of uPA , P05121 REA , and urokinase receptor showed substantial migration and invasion compared to other cell lines that lack expression of these proteins . However , overexpression of active wild-type P05121 REA , but not P14 - P05121 REA ( T333R ) , in these cell lines showed reduced migration and invasion . Since vitronectin binding by both forms of P05121 REA is equivalent , these results imply that P05121 REA - vitronectin interactions are less critical in altering migration and invasion . Our results show that the in vitro migratory and invasive phenotype in these breast and ovarian cancer cell lines is reduced by active P05121 REA due to its ability to inhibit plasminogen activation .

[ Treatment of renal vein thromboses in the newborn ] . Surgical thrombectomy is not a rational approach to neonatal renal vein thrombosis since the occlusion mainly involves intrarenal branches rather than the main renal vein , which is even patent in some instances . Conservative management combines supportive therapy for renal failure and systemic hypertension , if needed , and either heparin or thrombolytic agents . DB00086 has proven difficult to handle in neonates and should not be used . DB00013 SUB has been used in 18 patients but results are difficult to interpret because these cases occurred over an 18 - year period . P00747 REA tissue activator , the latest thrombolytic agent developed , has been used in few pediatric patients . An international task force is currently studying whether or not a randomized study is warranted to provide data for standardizing thrombolytic therapy in pediatric renal vein thrombosis .

DB00013 SUB plasminogen activator ( uPA ) and its receptor ( Q03405 REA ) in gestational tissues ; Measurements and clinical implications . BACKGROUND : DB00013 SUB plasminogen activator ( uPA ) and urokinase plasminogen activator receptor ( Q03405 REA ) are central molecules for uPA / Q03405 REA / plasmin-dependent proteolysis , which is thought to play a significant role in the development of pregnancy , as well as its many complications . OBJECTIVE : To measure the levels of uPA and Q03405 REA in the placenta and myometrium , as well as in the foetal membranes and amniotic fluid . STUDY DESIGN : The study group consisted of 35 women with normal course of pregnancy , but with complications arising during delivery , which led to Caesarean section . Samples of placenta , myometrium , foetal membranes , amniotic fluid and blood were obtained at the time of operation . Tissue extracts were prepared . Measurements were made by the ELISA method . RESULTS : uPA and Q03405 REA concentration in gestational tissues , including amniotic fluid , is 100-200 times higher than in plasma . Among tissues , the highest uPA level was found in placenta ( 1.32 + / - 0.48 ng / mg of protein ) , and the highest Q03405 REA level in foetal membranes ( 3.33 + / - 1.20 ng / mg of protein ) . CONCLUSIONS : uPA and Q03405 REA are present in all gestational tissues , in some in relatively high concentrations . Our results support the modern clinical hypothesis that fibrinolytic system can participate in mechanisms of such obstetric complications as pre-term pre-mature rupture of foetal membranes and placental abruption .

The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 REA activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor - 1 ( P05121 REA ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 REA activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline - and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai 1 - / - mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 REA ) and instead reflected proteolytic activation and release of P14210 REA with subsequent induction of P35354 REA . That the P14210 REA / P35354 REA / DB00917 axis mediates in vivo protection from fibrosis in Pai 1 - / - mice was demonstrated by experiments showing that a selective inhibitor of the P08581 REA c - DB00134 increased lung collagen to WT levels while reducing P35354 REA protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 REA and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 REA . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway .

DB11320 , carbachol , and serotonin induce hyperresponsiveness to DB00171 in guinea pig tracheas : involvement of P35354 REA pathway . Extracellular DB00171 promotes an indirect contraction of airway smooth muscle via the secondary release of thromboxane A2 ( TXA 2 ) from airway epithelium . Our aim was to evaluate if common contractile agonists modify this response to DB00171 . Tracheas from sensitized guinea pigs were used to evaluate DB00171 - induced contractions before and after a transient contraction produced by histamine , carbachol , or serotonin . Epithelial mRNA for P23219 REA and P35354 REA was measured by RT-PCR and their expression assessed by immunohistochemistry . Compared with the initial response , DB00171 - induced contraction was potentiated by pretreatment with histamine , carbachol , or serotonin . Either suramin ( antagonist of P2X and P2Y receptors ) plus Q08999 REA ( antagonist of P2Y receptors ) or indomethacin ( inhibitor of P23219 REA and P35354 REA ) annulled the DB00171 - induced contraction , suggesting that it was mediated by P2Y receptor stimulation and TXA 2 production . When P35354 REA was inhibited by SC - 58125 or thromboxane receptors were antagonized by SQ - 29548 , just the potentiation was abolished , leaving the basal response intact . Airway epithelial cells showed increased P35354 REA mRNA after stimulation with histamine or carbachol , but not serotonin , while P23219 REA mRNA was unaffected . Immunochemistry corroborated this upregulation of P35354 REA . In conclusion , we showed for the first time that histamine and carbachol cause hyperresponsiveness to DB00171 by upregulating P35354 REA in airway epithelium , which likely increases TXA 2 production . Serotonin-mediated hyperresponsiveness seems to be independent of P35354 REA upregulation , but nonetheless is TXA 2 dependent . Because acetylcholine , histamine , and serotonin can be present during asthmatic exacerbations , their potential interactions with DB00171 might be relevant in its pathophysiology .

Differential roles of Q03405 REA in peritoneal ovarian carcinomatosis . Epithelial ovarian cancer is the fourth leading cause of death from gynecologic malignancies in the United States . Most cases are diagnosed at late stages , with the solid tumor masses growing as peritoneal implants , or floating within the ascitic fluid ( peritoneal ovarian carcinomatosis ) . Despite aggressive surgical " debulking , " recurrence of recalcitrant disease is frequent with poor patient survival . Efforts to improve survival rates are hindered by lack of biomarkers that can detect and effectively treat ovarian cancer in its early stages . DB00013 SUB plasminogen activator receptor ( Q03405 REA ) is a multifunctional receptor involved in a myriad of tumor cell processes . However , the role of host Q03405 REA in ovarian cancer is still elusive . To define the potential proinflammatory role of Q03405 REA in ovarian cancer , first , using a syngeneic murine model in Q03405 REA ( - / - ) mice , we found that ablation of Q03405 REA restrained tumor take and peritoneal implants and prolonged the survival of Q03405 REA ( - / - ) mice compared with their Q03405 REA ( + / + ) counterparts . Ascitic fluid accumulation was significantly decreased in Q03405 REA ( - / - ) mice with decreased macrophage infiltration . Second , in vitro mechanistic studies revealed that host Q03405 REA is involved in the multiple steps of peritoneal metastatic cascade . Third , we evaluated the prognostic utility of tumor and stromal Q03405 REA in human ovarian cancer tissue microarray . In summary , our studies indicated that Q03405 REA plays a significant role in ovarian cancer cell-stromal crosstalk and contributes to increased vascular permeability and inflammatory ovarian cancer microenvironment . This provides a rationale for targeting the Q03405 REA with either specific neutralizing antibodies or targeting its downstream inflammatory effectors in patients with ovarian cancer .

DB00107 increases invasive properties of endometrial cancer cells through phosphatidylinositol 3 - kinase / AKT-dependent up-regulation of cyclooxygenase - 1 , - 2 , and P98170 REA . Traditionally , oxytocin ( OT ) is well known to play a crucial role in the regulation of cyclic changes in the uterus , implantation of the embryo , and parturition . Recently , an additional role for OT has been identified in several types of cancer cells in which OT acts as a growth regulator . In endometrial cancer cells , OT is known to efficiently inhibit cellular proliferation . In the present study , we show that OT increases invasiveness of human endometrial carcinoma ( O14777 REA ) cells , which are otherwise resistant to the growth-inhibiting effects of OT . Using pharmacological inhibitors , invasion assay , RNA interference , and immunofluorescence , we found that OT enhances the invasive properties of O14777 REA cells through up-regulation of P98170 REA ( P98170 REA ) , matrix-metalloproteinase 2 ( P08253 REA ) , and matrix-metalloproteinase 14 ( P50281 REA ) . In addition , we show that OT-mediated invasion is both cyclooxygenase 1 ( P23219 REA ) and cyclooxygenase - 2 ( P35354 REA ) dependent via the phosphatidylinositol 3 - kinase / AKT ( PIK 3 / AKT ) pathway . P35354 REA knockdown by shRNA resulted in P98170 REA down-regulation . We also show that OT receptor is overexpressed in grade I to III endometrial cancer . Taken together , our results describe for the first time a novel role for OT in endometrial cancer cell invasion .

Expression , purification , and biological characterization of the amino-terminal fragment of urokinase in Pichia pastoris . DB00013 SUB ( uPA ) and its receptor ( Q03405 REA ) play an important role in tumor growth and metastasis . Targeting the excessive activation of this system as well as the proliferation of the tumor vascular endothelial cell would be expected to prevent tumor neovasculature and halt the tumor development . In this regard , the amino-terminal fragment ( P39905 REA ) of urokinase has been confirmed as effective to inhibit the proliferation , migration , and invasiveness of cancer cells via interrupting the interaction of uPA and Q03405 REA . Previous studies indicated that P39905 REA expressed in Escherichia coli was mainly contained in inclusion bodies and also lacked posttranslational modifications . In this study , the biologically active and soluble P39905 REA was cloned and expressed in Pichia pastoris . The recombinant protein was purified to be homogenous and confirmed to be biologically active . The yield of the active P39905 REA was about 30 mg / l of the P . pastoris culture medium . The recombinant P39905 REA ( rATF ) could efficiently inhibit angiogenesis , endothelial cell migration , and tumor cell invasion in vitro . Furthermore , it could inhibit in vivo xenograft tumor growth and prolong the survival of tumor-bearing mice significantly by competing with uPA for binding to cell surfaces . Therefore , P . pastoris is a highly efficient and cost-effective expression system for large-scale production of biologically active rATFs for potential therapeutic application .

DB00013 SUB plasminogen activator activity is increased in the myocardium during coronary artery occlusion . We have previously demonstrated that collateral development takes place in a swine model of coronary artery occlusion . In this report we have examined the effect of coronary artery occlusion on urokinase and tissue plasminogen activator activity in the myocardium . DB00013 SUB activity was increased four-fold in the ischemic heart compared to sham and unoperated controls . In contrast , the level of tissue plasminogen activator activity remained relatively constant . The increase in urokinase activity was associated with an upregulation of urokinase RNA levels and of the RNAs corresponding to the plasminogen activator inhibitors , P05121 REA I and II . DB00013 SUB has been shown to be an important angiogenic protease both in vivo and in cultured cells . Its increase during collateral development suggests that urokinase may play a role in angiogenesis in the ischemic heart .

DB00013 SUB plasminogen activator induces angiogenesis and tumor vessel invasion in breast cancer . DB00013 SUB plasminogen activator ( uPA ) is a proteolytic enzyme implicated in cancer invasion and tumor progression . DB00013 SUB PA and its inhibitor ( P05121 REA ) appear to be new and independent prognostic markers in breast cancer . To investigate how uPA - and P05121 REA - levels correlate with angiogenesis and tumor vessel invasion , we counted microvessels and their tumor invasion and determined the uPA - and P05121 REA levels in 42 primary invasive breast carcinomas . 20 Patients had no lymph node metastasis at the time of surgery , while 22 patients had positive nodes . Using light microscopy , we highlighted the vessels by staining their endothelial cells immunocytochemically for CD31 and Factor VIII . After gaining tumor tissue extracts , we determined the uPA - and P05121 REA - levels by ELISA . A positive correlation between microvessel density , angioinvasion and uPA - and P05121 REA - levels was found . We speculate that high uPA levels may induce tumor neovascularisation , angioinvasion and may cause tumor progression and metastasis . The degradation of the vessel wall by uPA causes a leak . This wall defect may , on the one hand , be the stimulus for endothelial cell proliferation and formation of new blood vessels and , on the other hand , it may be the place of tumor cell entry .

Comparison of expression profiles induced by dust mite in airway epithelia reveals a common pathway . BACKGROUND : Airway epithelial cells have shown to be active participants in the defense against pathogens by producing signaling and other regulatory molecules in response to the encounter . METHODS : In previous manuscripts , we have studied the effect of house dust mite ( HDM ) extract on both an epithelial cell-line ( H292 ) and primary nasal epithelial cell . When we compare these responses we conclude that the H292 cells more closely resemble nasal epithelium of healthy controls ( share 107 probe-sets ) than of allergic individuals ( share 17 probe-sets ) . RESULTS : Interestingly , probably because of an absent intraindividual variation between samples , more probe-sets ( 8280 ) change expression significantly in H292 than in either healthy ( 555 ) or allergic ( 401 ) epithelium . CONCLUSIONS : A direct comparison of all the responses in these epithelial cells reveals a core-response to HDM of just 29 genes . These genes ( P78556 REA , P10145 REA , P19875 REA , P09341 REA , IL - 1B , P15514 REA , P21580 REA , Q99075 REA , P35354 REA , P12643 REA , P01130 REA , Q03405 REA , P00749 REA , Q00653 REA , P19838 REA , P05412 REA , P18847 REA , P18146 REA , O15118 REA , Q8IUC6 , P29317 REA , P29279 REA , P28562 REA , O43609 REA , TLR - 3 , complement factor P01024 REA , Q9Y6Y0 , SerpinB 3 , and Q9Y617 REA ) have described links with allergy or inflammation and may even describe the well-established relationship between viral infections and allergic exacerbations or allergy development .

Investigation of the binding of isoform-selective inhibitors to prostaglandin endoperoxide synthases using fluorescence spectroscopy . Prostaglandin endoperoxide synthase ( PGHS ) is a heme protein that catalyzes the committed step in prostaglandin and thromboxane biosynthesis . Two isoforms of PGHS exist , a constitutive form termed P23219 REA and an inducible form termed P35354 REA . We report here fluorescence resonance energy transfer analysis of isoform-selective inhibitors interacting with P23219 REA and P35354 REA . By measuring fluorescence quenching due to the energy transfer of the inhibitor fluorescence to the heme prosthetic group of PGHS , we determined these inhibitors bind in the arachidonic acid substrate access channel with an R0 of 35 A for P23219 REA with the P23219 REA inhibitor and an R0 of 21 A for P35354 REA with the P35354 REA inhibitor . The observed fluorescence quenching is completely dynamic and dominated by quenching by the heme . Time-resolved results combined with molecular modeling determine the distance from the inhibitor to the heme moiety to be 20 A in P23219 REA and 18 A in P35354 REA . Preliminary stopped-flow kinetic studies reveal that the rate of quenching is limited by a first-order protein transition , which is slow , and that bound inhibitor undergoes rapid exchange .

Down-regulation of Q03405 REA and uPA activates caspase-mediated apoptosis and inhibits the PI3K / AKT pathway . DB00013 SUB plasminogen activator ( uPA ) and its receptor ( Q03405 REA ) play a major role in invasion and proliferation . A growing body of evidence has suggested that the uPA system promotes tumor metastasis by several different mechanisms , and not just solely by breaking down the Q13201 REA . In this study we have used RNAi-mediated simultaneous down-regulation of Q03405 REA and uPA to determine the signaling pathway molecules and caspase-mediated apoptosis . From our in vitro experiments , we have observed that plasmid-based RNAi-mediated down-regulation of Q03405 REA and uPA in SNB 19 human glioma cells caused a decrease in the levels of Q03405 REA protein and uPA enzyme activities . In addition , we observed a decrease in the phosphorylation of the Ras-activated pathway molecules such as Q05397 REA , p38MAPK , JNK and P27361 REA / 2 , as well as the MEK-activated phosphatidylinositol 3 - kinase ( PI3k ) pathway , and also retarded the dephosphorylation of p-AKTser 473 and p-mTORser 2448 , indicative of a feedback signaling mechanism of the Q03405 REA - uPA system . Activation of caspase 8 accompanied by the release of cytochrome c and cleavage of PARP was also observed and indicative of Fas-mediated apoptosis . The use of FMK-VAD - Q05397 REA peptides coupled with FITC indicated activation of polycaspases , which was accompanied by the presence of fragmented nuclei . Our studies provide evidence for the presence of a feedback response of the Q03405 REA - uPA system indicative of the multifaceted role of Q03405 REA , and also the therapeutic potential of simultaneously targeting Q03405 REA and uPA in cancer patients .

Differential inhibition of soluble and cell surface receptor-bound single-chain urokinase by plasminogen activator inhibitor type 2 . A potential regulatory mechanism . DB00013 SUB ( u-PA ) - mediated cell surface plasminogen activation is required for cellular tissue invasion . This invasion occurs in environments rich in plasminogen activator inhibitors ( PAIs ) , which efficiently inhibit receptor-bound two-chain u-PA . Single-chain u-PA ( scu-PA ) was recently found to efficiently initiate cell surface plasminogen activation , and we herein describe the interaction of scu-PA with P05121 REA type 2 ( P05120 REA ) . In the fluid phase ( no cells ) the plasminogen-activating activities of both scu-PA and Glu 158 - scu-PA ( a plasmin non-activatable variant of scu-PA ) were inhibited in a concentration-dependent manner by recombinant human P05120 REA . This inhibition occurred with both forms of scu-PA remaining as single-chain molecules throughout the interactions . Although scu-PA did not form SDS-stable complexes with P05120 REA , preincubation of scu-PA with 125I - P05120 REA demonstrated a dose-dependent inhibition of SDS-stable complex formation between 125I - P05120 REA and subsequently added two-chain u-PA . This indicates that although a " stable intermediate " type complex between scu-PA and P05120 REA was not detected , there was a physical association between the two molecules that shared at least some determinants with the two-chain u-PA - P05120 REA complex . In contrast , Glu 158 - scu-PA bound to u-PA receptors on monocytes was only minimally inhibited by a large molar excess of P05120 REA . These data suggest that the initiation of cell surface plasminogen activation may involve the partitioning of scu-PA between P05120 REA ( a " negative modulator " ) and the u-PA receptor ( a " positive modulator " ) and that the enzymatic activity of receptor-bound scu-PA may allow initiation of cell surface proteolysis even in P05120 REA - rich environments . A model along these lines is presented .

DB01197 MEN attenuates matrix metalloproteinase - 2 and - 9 in monocrotaline-induced right ventricular hypertrophy in rats . Little is known about the influence of angiotensin converting enzyme ( P12821 REA ) inhibitors on matrix metalloproteinase ( MMP ) in right ventricular remodeling . We investigated the effect of captopril , an P12821 REA inhibitor , on P08253 REA and P14780 REA in monocrotaline-induced right ventricular hypertrophy . Six-week-old male Wistar rats were injected intraperitoneally with monocrotaline ( 60 mg / kg ) or saline . The rats were administrated captopril ( 30 mg / kg per day ) or a vehicle orally for 24 days from the day of monocrotaline injection . At day 25 , echocardiography was performed and hearts were excised . Expressions and activities of P08253 REA and P14780 REA were measured by Western blotting and by gelatin zymography , respectively . In monocrotaline-injected rats , right ventricular weight / tail length ratio increased significantly . Histological analysis revealed cardiomyocyte hypertrophy and fibrosis in right ventricular sections . Echocardiography showed right ventricular dysfunction compared with saline-injected rats . The right ventricular hypertrophy , fibrosis , and dysfunction were inhibited by captopril . However , captopril did not attenuate an increase in pulmonary artery pressure . P08253 REA and P14780 REA expressions and activities in right ventricles increased significantly in monocrotaline-injected rats and captopril inhibited them . These findings indicate that captopril attenuates the development of monocrotaline-induced right ventricular hypertrophy in association with inhibition of P08253 REA and P14780 REA in rats .

DB00013 SUB plasminogen activator is elevated in human astrocytic gliomas relative to normal adjacent brain . We assayed urokinase plasminogen activator ( uPA ) , tissue type plasminogen activator ( tPA ) , and plasminogen activator inhibitor - 1 ( P05121 REA ) in 43 human brain tumors ( predominantly astrocytic gliomas ) and in histologically disease-free brain tissue resected with 21 of the tumors . Levels of uPA , tPA , and P05121 REA , measured by enzyme-linked immunosorbent assay , varied widely among individuals in neoplastic and in normal tissue but did not correlate with age or sex . Pairwise comparison of neoplastic and normal tissue from 21 individuals revealed that mean tumor uPA level was elevated 6 - fold ( P < 0.001 ) . Mean tumor tPA and P05121 REA were 2.5- fold greater than those of normal brain , but these differences were not statistically significant . Tumor uPA was elevated 2 - to 30 - fold in 16/21 paired samples ( 76 % ) . In contrast , tumor tPA was elevated 2 - to 22 - fold in 7/21 ( 33 % ) of pairs , whereas tumor P05121 REA was 2 - to 13 - fold greater in 10/21 ( 48 % ) of pairs . Our results demonstrate that elevation of uPA content is frequent in astroglial tumors , as is the case in other major human cancers .

P00747 REA activator inhibitor - 1 ( P05121 REA ) and urokinase plasminogen activator ( uPA ) in sputum of allergic asthma patients . DB00013 SUB plasminogen activator ( uPA ) and its inhibitor ( P05121 REA ) have been associated with asthma . The aim of this study was to evaluate concentration of uPA and P05121 REA in induced sputum of house dust mite allergic asthmatics ( HDM-AAs ) . The study was performed on 19 HDM-AAs and 8 healthy nonatopic controls ( HCs ) . Concentration of uPA and P05121 REA was evaluated in induced sputum supernatants using ELISA method . In HDM-AAs the median sputum concentration of uPA ( 128 pg / ml ; 95 % CI 99 to 183 pg / ml ) and P05121 REA ( 4063 pg / ml ; 95 % CI 3319 to 4784 pg / ml ) were significantly greater than in HCs ( 17 pg / ml ; 95 % CI 12 to 32 pg / ml ; p < 0.001 and 626 pg / ml ; 95 % CI 357 to 961 pg / ml ; p < 0.001 for uPA and P05121 REA respectively ) . The sputum concentration of uPA correlated with sputum total cell count ( r = 0.781 ; p= 0.0001 ) and with logarithmically transformed exhaled nitric oxide concentration ( eNO ) ( r = 0.486 ; p= 0.035 ) but not with FEV 1 or bronchial reactivity to histamine . On the contrary , the sputum P05121 REA concentration correlated with FEV 1 ( r = -0,718 ; p= 0.0005 ) and bronchial reactivity to histamine expressed as log ( PC20 ) ( r = -0.824 ; p < 0.0001 ) but did not correlate with sputum total cell count or eNO . The results of this study support previous observations linking P05121 REA with airway remodeling and uPA with cellular inflammation . Moreover , the observed effect of uPA seems to be independent of its fibrynolytic activity .

O43609 REA promotes the degradation of Q03405 REA and inhibits Q03405 REA - mediated cell adhesion and proliferation . DB00013 SUB plasminogen activator receptor ( Q03405 REA ) is a P06744 REA anchored cell surface protein that is closely associated with invasion , migration , and metastasis of cancer cells . Many functional extracellular proteins and transmembrane receptors interact with Q03405 REA . However , few studies have examined the association of Q03405 REA with cytoplasm proteins . We previously used yeast two-hybrid screening to isolate several novel Q03405 REA - interacting cytoplasmic proteins , including Sprouty 1 ( O43609 REA ) , an inhibitor of the ( Ras-mitogen-activated protein kinase ) MAPK pathway . In this study , we show that O43609 REA interacts with Q03405 REA and directs it toward lysosomal-mediated degradation . Overexpression of O43609 REA decreased the cell surface and cytoplasmic Q03405 REA protein level . Moreover , O43609 REA overexpression augmented Q03405 REA - induced cell adhesion to vitronectin as well as proliferation of cancer cells . Our results also further support the critical role of O43609 REA contribution to tumor growth . In a subcutaneous tumor model , overexpression of O43609 REA in HCT 116 or A549 xenograft in athymic nude mice led to great suppression of tumor growth . These results show that O43609 REA may affect tumor cell function through direct interaction with Q03405 REA and promote its lysosomal degradation .

The urokinase-type plasminogen activator receptor mediates tyrosine phosphorylation of focal adhesion proteins and activation of mitogen-activated protein kinase in cultured endothelial cells . P00749 REA ( uPA ) binds to cells via a specific glycosylphosphatidylinositol-anchored receptor . Although occupancy of the uPA receptor ( Q03405 REA ) has been shown to alter cellular function and to induce gene expression , the signaling mechanism has not been characterized . DB00013 SUB induced an increase in the tyrosine phosphorylation of multiple proteins in bovine aortic endothelial cells . In contrast , low molecular weight uPA did not induce this response . Analysis by immunoblotting demonstrated tyrosine phosphorylation of focal adhesion kinase ( Q05397 REA ) , the focal adhesion-associated proteins paxillin and Q08999 REA ( cas ) , and mitogen-activated protein kinase ( MAPK ) following the occupancy of the Q03405 REA by uPA . Treatment of cells with phosphatidylinositol-specific phospholipase C , which cleaves glycosylphosphatidylinositol-linked proteins from the cell surface , blocked the uPA-induced tyrosine phosphorylation of Q05397 REA , indicating the requirement of an intact Q03405 REA on the cell surface . The uPA-induced activation of MAPK was completely inhibited by genistein , but not by 4 - amino - 5 - ( 4 - methylphenyl ) - 7 - ( t-butyl ) pyrazolo [ 3 , 4 - d ] pyrimidine , a specific inhibitor of Src family kinases . Thus , this study demonstrates a novel role for the Q03405 REA in endothelial cell signal transduction that involves the activation of Q05397 REA and MAPK , which are mediated by the receptor-binding domain of uPA . This may have important implications for the mechanism through which uPA influences cell migration and differentiation .

DB00013 SUB induces matrix metalloproteinase - 9 / gelatinase B expression in THP - 1 monocytes via P27361 REA / 2 and cytosolic phospholipase A2 activation and eicosanoid production . OBJECTIVE : P00749 REA ( uPA ) regulates cell migration and invasion by pericellular proteolysis and signal transduction events . We characterized the mechanisms by which uPA regulates matrix metalloproteinase - 9 ( P14780 REA ) function in THP - 1 monocytes . METHODS AND RESULTS : In THP - 1 monocytes , P14780 REA production induced by urokinase was completely inhibited by the P27361 REA / 2 inhibitor , PD98059 , but not by the p38 mitogen-activated protein kinase inhibitor , SB202190 . A dominant negative Q02750 REA adenovirus also blocked P14780 REA expression . The effect of urokinase was completely suppressed by genistein and by herbimycin A indicating that tyrosine kinase ( s ) are required for P14780 REA production . Bisindolylmaleimide , a protein kinase C ( PKC ) inhibitor , did not decrease P14780 REA expression suggesting that PKC activation is not required . Key roles for cytosolic phospholipase A2 ( P04054 REA ) and eicosanoid production were shown by complete inhibition with methyl arachidonyl fluorophosphonate ( an inhibitor of cytosolic P04054 REA ) , and indomethacin ( a cyclooxygenase inhibitor ) , with no effect of monoalide , a secretory P04054 REA inhibitor . uPA stimulated phosphorylation of cytosolic P04054 REA . CONCLUSIONS : Induction of P14780 REA by uPA in THP - 1 monocytes is via a pathway involving Q02750 REA - P27361 REA / 2 - mediated activation of cytosolic P04054 REA and eicosanoid generation . These data suggest important roles for eicosanoids in monocyte migration induced by uPA and P14780 REA .

DB00013 SUB receptor primes cells to proliferate in response to epidermal growth factor . Epidermal growth factor ( P01133 REA ) expresses mitogenic activity by a mechanism that requires the P01133 REA receptor ( P00533 REA ) . We report that murine embryonic fibroblasts ( MEFs ) proliferate in response to P01133 REA only when these cells express the urokinase receptor ( Q03405 REA ) . P00533 REA expression was equivalent in Q03405 REA - / - and Q03405 REA + / + MEFs . In response to P01133 REA , these cells demonstrated equivalent overall P00533 REA tyrosine phosphorylation and P29323 REA / Q96HU1 kinase activation ; however , phosphorylation of DB00135 - 845 in the P00533 REA , which has been implicated in cell growth , was substantially decreased in Q03405 REA - / - MEFs . STAT 5b activation also was decreased . As DB00135 - 845 is a c-Src target , we overexpressed c-Src in Q03405 REA - / - MEFs and rescued P01133 REA mitogenic activity . Rescue also was achieved by expressing murine but not human Q03405 REA , suggesting a role for autocrine Q03405 REA cell-signaling . In MDA-MB 231 breast cancer cells , P01133 REA mitogenic activity was blocked by Q03405 REA gene silencing , with antibodies that block uPA-binding to Q03405 REA , and with a synthetic peptide that disrupts Q03405 REA - dependent cell signaling . Again , c-Src overexpression rescued the mitogenic activity of P01133 REA . We conclude that Q03405 REA - dependent cell-signaling may prime cells to proliferate in response to P01133 REA by promoting DB00135 - 845 phosphorylation and STAT 5b activation . The importance of this pathway depends on the c-Src level in the cell .

Emodin upregulates urokinase plasminogen activator , plasminogen activator inhibitor - 1 and promotes wound healing in human fibroblasts . DB00013 SUB plasminogen activator ( uPA ) system is important for several biological processes that call for extracellular proteolysis , fibrinolysis , cell migration , proliferation and angiogenesis . The current study highlights the fibrinolytic and wound healing potential of emodin , an anthraquinone , with relevance to the uPA system . Emodin increased the fibrinolytic activity of fibroblast cells in a dose-dependent manner . Zymography linked the activity to increased uPA activity . Subsequent RT-PCR and western analyses demonstrated uPA gene upregulation . Interestingly , P05121 REA , the inhibitor of uPA was also upregulated . EMSA showed the upregulation occurred independent of emodin ' s effect on nuclear factor kappa B ( NFkappaB ) . The effect on uPA system is supposedly via generation of reactive oxygen species ( ROS ) since cotreatment with ascorbic acid , an anti-oxidant , attenuated the activity . In addition to profibrinolytic potential , emodin also demonstrated wound healing activity in in vitro wound models . Presence of emodin in the medium enhanced the rate of migration of fibroblasts into the wounded region . These in vitro experiments reveal that emodin is a potent profibrinolytic and wound healing agent .

P35354 REA induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways . The functional significance of protease-activated receptors ( PARs ) in endothelial cells is largely undefined , and the intracellular consequences of their activation are poorly understood . Here , we show that the serine protease thrombin , a P25116 REA - selective peptide ( TFLLRN ) , and SLIGKV ( P55085 REA - selective peptide ) induce cyclooxygenase - 2 ( P35354 REA ) protein and mRNA expression in human endothelial cells without modifying P23219 REA expression . P35354 REA induction was accompanied by sustained production of 6 - keto-PGF 1alpha , the stable hydrolysis product of prostacyclin , and this was inhibited by indomethacin and the P35354 REA - selective inhibitor NS398 . P25116 REA and P55085 REA stimulation rapidly activated both P27361 REA / 2 and p38MAPK , and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin - and SLIGKV-induced P35354 REA expression and 6 - keto-PGF 1alpha formation . Thrombin and peptide agonists of P25116 REA and P55085 REA increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus , and this , as well as PAR-induced 6 - keto-PGF 1alpha synthesis , was inhibited by co-infection with adenovirus encoding wild-type or mutated ( Y42F ) P25963 REA . Thrombin - and SLIGKV-induced P35354 REA expression and 6 - keto-PGF 1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG - 132 . Activation of P25116 REA or P55085 REA promoted nuclear translocation and phosphorylation of p65 - NF-kappaB , and thrombin-induced but not P55085 REA - induced p65 - NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK . Activation of Q96RI0 by AYPGKF increased phosphorylation of P27361 REA / 2 and p38MAPK without modifying NF-kappaB activation or P35354 REA induction . Our data show that P25116 REA and P55085 REA , but not Q96RI0 , are coupled with P35354 REA expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on P27361 REA / 2 , p38MAPK , and P25963 REA - dependent NF-kappaB activation .

Regulation of the Q03405 REA / uPA system expressed on monocytes by the deactivating cytokines , P05112 REA , P22301 REA and P35225 REA : consequences on cell adhesion to vitronectin and fibrinogen . DB00013 SUB ( uPA ) and its receptor ( Q03405 REA ) have been proposed to be involved in monocyte migration by inducing degradation of matrix proteins . In addition , Q03405 REA is also implicated in cell adhesion to the vascular wall . The adhesive function of Q03405 REA depends on a direct interaction with vitronectin which is increased by uPA and by modification of cell surface integrin ( such as CD11b - P05107 REA ) when associated to Q03405 REA . In this study we analysed the role of three deactivating cytokines , P05112 REA , P22301 REA and P35225 REA , on the surface expression of uPA , Q03405 REA and CD11b by monocytes and their consequences on monocyte adhesion to immobilized fibrinogen and vitronectin . P22301 REA induced a decrease in uPA and CD11b after 18 h incubation and a delayed decrease in Q03405 REA which was only significant after 48 h incubation . These results may explain the decrease in monocyte adhesion , which was observed after an 18 h incubation with P22301 REA , on immobilized vitronectin and fibrinogen . In contrast , P05112 REA and P35225 REA induced a decrease in Q03405 REA after 18 h and a significant increase in uPA both in the cell lysates and at the cell surface , as well as an increase in cell surface associated CD11b . These cytokines did not modify cell adhesiveness to vitronectin or fibrinogen despite the increase in CD11b - P05107 REA . This could be due to the decrease in Q03405 REA because CD11b - P05107 REA / Q03405 REA forms a cell adhesion complex . In addition , the increase in uPA induced by P05112 REA could counterbalance the direct interaction of Q03405 REA with vitronectin . The increase in uPA suggests that P05112 REA and P35225 REA could induce plaque fissuring by monocytes , whereas P22301 REA may induce protection against matrix protein degradation by decreasing uPA .

DB00013 SUB receptor is necessary for adequate host defense against pneumococcal pneumonia . Cell recruitment is a multistep process regulated by cytokines , chemokines , and growth factors . Previous work has indicated that the urokinase plasminogen activator receptor ( Q03405 REA ) may also play a role in this mechanism , presumably by an interaction with the beta ( 2 ) integrin CD11b / P05107 REA . Indeed , an essential role of Q03405 REA in neutrophil recruitment during pulmonary infection has been demonstrated for beta ( 2 ) integrin-dependent respiratory pathogens . We investigated the role of Q03405 REA and urokinase plasminogen activator ( uPA ) during pneumonia caused by a beta ( 2 ) integrin-independent respiratory pathogen , Streptococcus pneumoniae . Q03405 REA - deficient ( Q03405 REA ( - / - ) ) , uPA-deficient ( uPA ( - / - ) ) , and wild-type ( Wt ) mice were intranasally inoculated with 10 ( 5 ) CFU S . pneumoniae . Q03405 REA ( - / - ) mice showed reduced granulocyte accumulation in alveoli and lungs when compared with Wt mice , which was associated with more S . pneumoniae CFU in lungs , enhanced dissemination of the infection , and a reduced survival . In contrast , uPA ( - / - ) mice showed enhanced host defense , with more neutrophil influx and less pneumococci in the lungs compared with Wt mice . These data suggest that Q03405 REA is necessary for adequate recruitment of neutrophils into the alveoli and lungs during pneumonia caused by S . pneumoniae , a pathogen eliciting a beta ( 2 ) integrin-independent inflammatory response . This function is even more pronounced when Q03405 REA is unoccupied by uPA .

DB00013 SUB induces proliferation of human ovarian cancer cells : characterization of structural elements required for growth factor function . Ovarian cancer metastasis is associated with an increase in the urokinase-type plasminogen activator ( uPA ) and its receptor Q03405 REA . We present evidence that binding of uPA to Q03405 REA provokes a mitogenic response in the human ovarian cancer cell line OV-MZ - 6 in which endogenous uPA production had been significantly reduced by stable uPA ' antisense ' transfection . High molecular weight ( HMW ) uPA , independent of its enzymatic activity , produced an up to 95 % increase in cell number concomitant with 2 - fold elevated [ 3H ] thymidine incorporation as did the catalytically inactive but Q03405 REA binding amino-terminal fragment of uPA , P39905 REA . uPA-induced cell proliferation was significantly decreased by blocking uPA / Q03405 REA interaction by the monoclonal antibody IIIF 10 and by soluble Q03405 REA . The efficiency of the Q03405 REA binding synthetic peptide cyclo 19,31 uPA 19-31 to enhance OV-MZ - 6 cell growth proved this molecular domain to be the minimal structural determinant for uPA mitogenic activity . Dependence of uPA-provoked cell proliferation on Q03405 REA was further demonstrated in Raji cells which do not express Q03405 REA and were thus not induced by uPA . However , upon transfection with full-length Q03405 REA , Raji cells acquired a significant growth response to HMW uPA and P39905 REA .

P00749 REA in endothelial cells during acute inflammation of the appendix . DB00013 SUB and tissue-type plasminogen activators ( u-PA and t-PA ) were identified immunohistochemically in normal and inflamed human appendices by means of polyclonal and monoclonal antibodies . In addition , extracts of the tissues were analyzed for u-PA and t-PA by ELISA . Twelve appendices ( five normal and seven with acute inflammation ) were analyzed . In the normal appendices , there was a strong staining of the endothelial cells for t-PA , whereas there was negative staining for u-PA . In contrast , the endothelial cells in the inflamed appendices showed u-PA immunoreactivity , and negative or very weak reactions for t-PA . In the inflamed appendix , there was also a labeling of u-PA in fibroblast-like cells and in interstitial areas . The specificity of the staining was supported by a variety of staining controls and also by analysis of tissue extracts with ELISA , showing that on the average the inflamed appendices contained more than twice as much mu-PA per mg of protein as the normal appendices and less than one third of the amount of t-PA .

Targeting of tumor cells by cell surface urokinase plasminogen activator-dependent anthrax toxin . DB00013 SUB plasminogen activator receptor ( Q03405 REA ) binds pro-urokinase plasminogen activator ( pro-uPA ) and thereby localizes it near plasminogen , causing the generation of active uPA and plasmin on the cell surface . Q03405 REA and uPA are overexpressed in a variety of human tumors and tumor cell lines , and expression of Q03405 REA and uPA is highly correlated to tumor invasion and metastasis . To exploit these characteristics in the design of tumor cell-selective cytotoxins , we constructed mutated anthrax toxin-protective antigen ( PrAg ) proteins in which the furin cleavage site is replaced by sequences cleaved specifically by uPA . These uPA-targeted PrAg proteins were activated selectively on the surface of Q03405 REA - expressing tumor cells in the presence of pro-uPA and plasminogen . The activated PrAg proteins caused internalization of a recombinant cytotoxin , FP59 , consisting of anthrax toxin lethal factor residues 1-254 fused to the ADP-ribosylation domain of Pseudomonas exotoxin A , thereby killing the Q03405 REA - expressing tumor cells . The activation and cytotoxicity of these uPA-targeted PrAg proteins were strictly dependent on the integrity of the tumor cell surface-associated plasminogen activation system . We also constructed a mutated PrAg protein that selectively killed tissue plasminogen activator-expressing cells . These mutated PrAg proteins may be useful as new therapeutic agents for cancer treatment .

Regulation and role of urokinase plasminogen activator in vascular remodelling . 1 . DB00013 SUB plasminogen activator ( uPA ) is produced and secreted by multiple vascular cell types , thus influencing the processes and the extent to which the vasculature is remodelled during the development of the intima or a neointima and during hypertrophy and angiogenesis . 2 . DB00013 SUB plasminogen activator mRNA expression is up - and down-regulated by growth factors , cytokines and steroids . DB00013 SUB plasminogen activator is secreted as a single chain inactive form that may be proteolytically converted to active or inactive forms . Targeting of proteolytic activity may occur via focalized expression of uPA and its cell surface receptors ( Q03405 REA ) . Proteolytic activity is also controlled through the often co-ordinated expression of specific inhibitors . 3 . A proteolytic cascade involving uPA provides its major role in tissue remodelling through the primary degradation of extracellular matrix and secondarily through the activation of transforming growth factor-beta or release from the matrix of basic fibroblast growth factor . In addition , uPA secreted by growth factor-stimulated vascular cells may contribute to the chemotactic and mitogenic responses ascribed to the growth factor and recent evidence strongly suggests that uPA has direct biological actions on vascular cells . 4 . The cell surface binding of uPA via its growth factor-like domain to Q03405 REA localizes and activates the protease , but may also initiate transmembrane signalling of biological responses , including migration / invasion and proliferation . As the Q03405 REA lacks intracellular signalling domains , the signals may be transduced via interactions between uPA / Q03405 REA and more classical signalling receptors . The mechanism by which uPA may be involved in cell signalling is yet to be elucidated .

Molecular imaging of pancreatic cancer in an animal model using targeted multifunctional nanoparticles . BACKGROUND & AIMS : Identification of a ligand / receptor system that enables functionalized nanoparticles to efficiently target pancreatic cancer holds great promise for the development of novel approaches for the detection and treatment of pancreatic cancer . DB00013 SUB plasminogen activator receptor ( Q03405 REA ) , a cellular receptor that is highly expressed in pancreatic cancer and tumor stromal cells , is an excellent surface molecule for receptor-targeted imaging of pancreatic cancer using multifunctional nanoparticles . METHODS : The Q03405 REA - targeted dual-modality molecular imaging nanoparticle probe is designed and prepared by conjugating a near-infrared dye-labeled amino-terminal fragment of the receptor binding domain of urokinase plasminogen activator to the surface of functionalized magnetic iron oxide nanoparticles . RESULTS : We have shown that the systemic delivery of Q03405 REA - targeted nanoparticles leads to their selective accumulation within tumors of orthotopically xenografted human pancreatic cancer in nude mice . The Q03405 REA - targeted nanoparticle probe binds to and is subsequently internalized by Q03405 REA - expressing tumor cells and tumor-associated stromal cells , which facilitates the intratumoral distribution of the nanoparticles and increases the amount and retention of the nanoparticles in a tumor mass . Imaging properties of the nanoparticles enable in vivo optical and magnetic resonance imaging of Q03405 REA - elevated pancreatic cancer lesions . CONCLUSIONS : Targeting Q03405 REA using biodegradable multifunctional nanoparticles allows for the selective delivery of the nanoparticles into primary and metastatic pancreatic cancer lesions . This novel receptor-targeted nanoparticle is a potential molecular imaging agent for the detection of pancreatic cancer .

DB00013 SUB / urokinase receptor and vitronectin / alpha ( v ) beta ( 3 ) integrin induce chemotaxis and cytoskeleton reorganization through different signaling pathways . P04004 REA ( VN ) and pro-urokinase ( pro-uPA ) stimulated migration of rat smooth muscle cells in a dose-dependent and additive way , and induced motile-type changes in cell morphology together with a complete reorganization of the actin filaments and of the microtubules . All these effects were inhibited by pertussis toxin , or by antibodies directed against the urokinase receptor ( Q03405 REA ) or against the VN receptor alpha ( v ) beta ( 3 ) suggesting that an association between the two receptors is required to mediate both signals . Investigation of the signaling pathways showed that increasing the intracellular DB02527 resulted in a selective inhibition of VN-induced cell migration . On the other hand , PD 98059 , an inhibitor of MEK , differentially inhibited the pro-uPA - but not the VN-induced cell migration . Phosphorylation and nuclear translocation of Erk by pro-uPA was directly observed . We conclude that the signaling pathways of pro-uPA and VN must be at least in part different .

P07996 REA and transforming growth factor beta - 1 upregulate plasminogen activator inhibitor type 1 in pancreatic cancer . Controlled degradation of the extracellular matrix by proteases is crucial in tumor cell invasion . We have shown that thrombospondin - 1 ( P07996 REA - 1 ) , through activation of transforming growth factor beta - 1 ( TGF-beta 1 ) , regulates the plasminogen / plasmin protease system in breast cancer . To determine whether this occurred in other epithelial neoplasms , we studied the role of P07996 REA - 1 and TGF-beta 1 in the regulation of the plasminogen / plasmin system in pancreatic cancer . ASPC - 1 and COLO - 357 pancreatic cancer cells were treated with P07996 REA - 1 or TGF-beta 1 at varying concentrations . The P07996 REA - 1 and TGF-beta 1 - treated cells were also treated with either anti - P07996 REA - 1 , anti - P07996 REA - 1 receptor , or anti-TGF-beta 1 antibodies . DB00013 SUB plasminogen activator ( uPA ) and plasminogen activator inhibitor - 1 ( P05121 REA ) expression was determined by enzyme-linked immunosorbent assay . P07996 REA - 1 and TGF-beta 1 promoted a dose-dependent upregulation of ASPC - 1 and COLO - 357 P05121 REA expression . The P07996 REA - 1 effect could be blocked with anti - P07996 REA - 1 or anti-TGF-beta 1 antibodies . The TGF-beta 1 effect could be blocked only with anti-TGF-beta 1 antibody . Anti - P07996 REA - 1 receptor antibody blocked the P07996 REA - 1 effect on P05121 REA expression but had no effect on TGF-beta 1 - mediated P05121 REA expression . Neither P07996 REA - 1 nor TGF-beta 1 had an effect on uPA production . We conclude that P07996 REA - 1 , in a receptor-mediated process that involves the activation of TGF-beta 1 , upregulates P05121 REA expression in pancreatic cancer without an effect on uPA production .

P00747 REA activator production in a rat model of Pneumocystis carinii pneumonia . Several studies have indicated that the serine protease urokinase-plasminogen-activator ( uPA ) is an important factor in host defense against pulmonary pathogens . To gain a better insight into the role of uPA in Pneumocystis carinii ( P . carinii ) pneumonia ( PCP ) , we evaluated PA production in alveolar macrophages ( AMs ) obtained from rats with steroid-induced PCP . Treatment with cortisone acetate favored PCP in 91 % of rats . In the bronchoalveolar lavage ( BAL ) samples of immunosuppressed rats both with and without PCP , we observed a decrease in uPA activity as well as a decrease in cell number . DB00013 SUB - PA production by AMs was reduced in rats treated with cortisone alone . However , an increase in cell-associated uPA was observed in rats with PCP . This increase appears to be produced in response to P carinii infection . In fact , when AMs obtained from untreated healthy or immunosuppressed uninfected rats were challenged with P carinii , a significant increase in PA activity in cell lysates was observed , though a lower response was obtained in cortisone-treated animals . Our results suggest that healthy AMs respond to the presence of P carinii with an increase in uPA production and that this response in immunodepressed rat-AMs is partially impaired .

P00747 REA activators and plasmin in lung cancer . DB00013 SUB plasminogen activator and plasmin contribute to detach neoplastic cells from solid tumor and facilitate the movement of these cells through interstitium and capillary walls as well as infiltration of the surrounding structures . P00747 REA activators inhibitors fulfill a regulatory function in these processes . Determining activity and concentration , finding subcellular , cellular and zonal localization of every component of plasminogen activation system has diagnostic and prognostic importance in different lung cancer types .

DB01197 MEN reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 REA ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 REA ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 REA inhibitor captopril reduces plasma P05121 REA inhibitor activity , we measured changes in plasma P05121 REA activity ( IU / ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng / ml ) , and serum P12821 REA activity ( IU / L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 REA activity decreased significantly , from 14.0 + / - 0.8 to 11.5 + / - 1.2 IU / L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 REA activity and t-PA antigen also decreased significantly-from 11.9 + / - 2.8 to 5.5 + / - 2.2 IU / ml ( p < 0.02 ) and from 9.9 + / - 1.0 to 7.5 + / - 0.9 ng / ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 REA activity , P05121 REA activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity .

P00533 REA is a transducer of the urokinase receptor initiated signal that is required for in vivo growth of a human carcinoma . DB00013 SUB plasminogen activator receptor ( Q03405 REA ) activates alpha 5beta1 integrin and P29323 REA signaling , inducing in vivo proliferation of HEp 3 human carcinoma . Here we demonstrate that P00533 REA mediates the Q03405 REA / integrin / fibronectin ( FN ) induced growth pathway . Its activation is ligand-independent and does not require high P00533 REA , but does require high Q03405 REA expression . Only when Q03405 REA level is constitutively elevated does P00533 REA become alpha 5beta1 - associated and activated . Domain 1 of Q03405 REA is crucial for P00533 REA activation , and Q05397 REA links integrin and P00533 REA signaling . Inhibition of P00533 REA kinase blocks Q03405 REA induced signal to P29323 REA , implicating P00533 REA as an important effector of the pathway . Disruption of Q03405 REA or P00533 REA signaling reduces HEp 3 proliferation in vivo . These findings unveil a mechanism whereby Q03405 REA subverts ligand-regulated P00533 REA signaling , providing cancer cells with proliferative advantage .

P00749 REA and plasminogen activator inhibitor type - 1 mRNA assessment in breast cancer by means of NASBA : correlation with protein expression . DB00013 SUB plasminogen activator ( uPA ) and its main inhibitor , plasminogen activator inhibitor type - 1 ( P05121 REA ) determined in tumor tissue by means of enzyme-linked immunosorbent assay ( ELISA ) can discriminate patients with primary breast cancer at high risk vs low risk for recurrence . The aim of this study was to analyze uPA and P05121 REA messenger RNA ( mRNA ) expression by means of quantitative nucleic acid sequence-based amplification ( NASBA ) on 77 primary breast tumor samples and to correlate this expression with the uPA and P05121 REA protein content . We observed that the 2 markers were significantly overexpressed ( uPA , P < . 0001 ; P05121 REA , P = . 0042 ) in mRNA in the ELISA + group . The receiver operating characteristic ( ROC ) curves demonstrated high concordance between NASBA and ELISA ( area under the ROC curve of 0.84 and 0.70 for uPA and P05121 REA , respectively ) and showed that uPA and P05121 REA status could be predicted by using the molecular assay with sensitivity and specificity values of 80.8 % and 82.4 % and sensitivity and specificity values of 66.7 % and 74.0 % , respectively .

P12821 REA inhibition suppresses plasminogen activator inhibitor - 1 expression in the neointima of balloon-injured rat aorta . BACKGROUND : P00747 REA activator inhibitor - 1 ( P05121 REA ) , an important regulator of fibrinolysis and extracellular matrix turnover , has been implicated in a number of vascular diseases . Studies demonstrating angiotensin II ( Ang II ) to be a potent stimulator of P05121 REA expression in cultured vascular cells suggests that the renin-angiotensin system may modulate vascular P05121 REA expression . METHODS AND RESULTS : We examined the effects of the P12821 REA inhibitor captopril on P05121 REA expression in control and balloon-injured rat aorta . Northern blot analysis demonstrated that aortic P05121 REA mRNA expression was 7.6- fold elevated 3 hours ( P < . 05 ) after balloon injury , back to baseline at 2 days , increased again at 4 days , and by 7 days after balloon injury was 3.2- fold elevated ( P < . 05 ) when compared with control . In captopril-treated rats , the induction of P05121 REA expression by balloon injury was significantly suppressed by 44 % ( P < . 05 ) in the 7 day group but was not altered in the 3 - hour group . DB01197 MEN also reduced baseline aortic P05121 REA mRNA . In situ hybridization and immunohistochemistry revealed dense P05121 REA staining of 7 - day neointima in untreated rats and a dramatic decrease in P05121 REA in neointima of captopril-treated rats . CONCLUSIONS : This report demonstrates that balloon injury results in both a rapid P12821 REA inhibitor-independent induction of aortic P05121 REA expression and a later increase in P05121 REA in the neointima that is significantly suppressed by captopril . This provides the first evidence that the renin-angiotensin system regulates neointimal P05121 REA expression and that P12821 REA inhibitors can reduce P05121 REA in the vessel wall in vivo .

DB00013 SUB plasminogen activator and its inhibitor , P05121 REA , as prognostic markers in breast cancer : from pilot to level 1 evidence studies . BACKGROUND : For optimum management of patients with cancer , accurate assessment of prognosis is essential . The primary determinant of outcome in malignancy is the formation of distant metastases . DB00013 SUB plasminogen activator ( uPA ) is a serine protease causally involved in invasion and metastasis . CONTENT : Data from model systems show that uPA is unequivocally involved in cancer dissemination . Consistent with its role in metastasis , multiple independent groups have shown that high uPA concentrations in primary breast cancers correlate with poor prognosis . For determining outcome , the prognostic impact of uPA was both independent of traditionally used factors and prognostic in patients with axillary node-negative disease . Paradoxically , high concentrations of plasminogen activator inhibitor ( P05121 REA ) , an endogenous inhibitor of uPA , also correlate with poor prognosis in patients with breast cancer , including the subgroup with node-negative disease . The prognostic value of uPA / P05121 REA in axillary node-negative breast cancer patients was recently confirmed in both a prospective randomized trial and a pooled analysis , i . e . , two different level 1 evidence ( LOE - 1 ) studies . CONCLUSIONS : uPA and P05121 REA are among the first biological prognostic factors to have their clinical value validated using LOE - 1 evidence studies . Determination of these analytes may help identify low-risk node-negative breast cancer patients for whom adjuvant chemotherapy is unnecessary .

DB00013 SUB plasminogen activator amino-terminal peptides inhibit development of the rat ventral prostate . The plasma membrane urokinase plasminogen activator receptor ( Q03405 REA ) localizes and enhances activation of pro-uPA . Active uPA , in turn , promotes increased degradation of the extracellular matrix ( Q13201 REA ) by activation of plasminogen . Q03405 REA binds to Q13201 REA molecules and integrins , which can affect cellular adhesion , signal transduction , and gene regulation . The current study examines the expression and function of Q03405 REA in developing rat ventral prostates ( VPs ) . We report that newborn VPs express Q03405 REA mRNA and protein . In addition , the function of Q03405 REA - bound uPA during in vitro prostatic development was studied by adding recombinant peptide competitive inhibitors of uPA - Q03405 REA binding . Newborn VP explants were cultured in serum-free media for one week with 10 (-8 ) M testosterone plus chimeric peptides containing a human immunoglobulin G Fc domain and either human uPA amino acids 1-138 ( hu-uPA 1-138 ) as a control or mouse uPA amino acids 1-138 ( mo-uPA 1-138 ) or 1-48 ( mo-uPA 1-48 ) . Hu-uPA 1-138- treated VPs underwent normal ductal branching morphogenesis and tissue differentiation . In contrast , VPs treated with mo-uPA 1-138 or mo-uPA 1-48 displayed a dose-dependent perturbation of ductal branching . Differentiation of both epithelial and mesenchymal tissues was also impaired . Mo-uPA 1-48- treated VPs contained significantly more apoptotic cells . These observations suggest that disruption of uPA binding to Q03405 REA results in a retardation of the development of newborn VPs .

DB00013 SUB plasminogen activator receptor : Prognostic biomarker for endometrial cancer . Endometrial adenocarcinoma is the most common gynecologic malignancy in the United States . However , reliable diagnostic or prognostic tumor markers have not been identified for endometrial cancer . In this study , we examined whether urokinase plasminogen activator receptor ( Q03405 REA ) , a glycosyl-phosphatidylinositol-linked membrane protein , is a candidate diagnostic or prognostic marker for patients with cancer of the endometrium . Sixty-five surgically excised , formalin-fixed endometrial tissue specimens were accessioned through the Department of Pathology Registry at the University of California , Los Angeles , and analyzed for Q03405 REA expression by using immunohistochemical techniques . A retrospective review was also performed to determine stage and histopathologic grade of disease , recurrence , and mortality . No expression of Q03405 REA protein was present in seven patients with benign neoplasia of the endometrium . Q03405 REA protein expression highly correlated with stage of disease ( ungrouped Spearman correlation = 0.625 , P < 0.0001 ) : 40 % of patients with stage I , 66 % of patients with stage II , 100 % of patients with stage III , and 85 % with stage IV demonstrated the highest level of Q03405 REA expression . Moreover , high Q03405 REA expression positively correlated with grade of disease ( ungrouped Spearman correlation = 0.71 , P < 0.0001 ) : 29 % of grade 1 specimens , 57 % of grade 2 , and over 90 % of specimens with grade 3 , the majority representing uterine papillary serous carcinoma and mixed malignant mesodermal tumor . Finally , Q03405 REA protein expression also positively correlated with rate of recurrence and mortality in patients with adenocarcinoma of the endometrium ( ungrouped P = 0.034 ) . Our data suggest that Q03405 REA is a useful prognostic marker for biologically aggressive forms of endometrial cancer .

Identification of molecular forms of plasminogen and plasmin-inhibitor complexes in urokinase-activated human plasma . DB00013 SUB - activated human plasma was analysed by acetic acid / urea / polyacrylamide-gel electrophoresis . The bands representing plasminogen , the plasmin-alpha 2 - plasmin inhibitor and plasmin-alpha 2 - macroglobulin complexes were identified by immunoprecipitation with specific antibodies and by comparison with purified components . P00747 REA and the plasmin-inhibitor complexes were isolated from plasma or thrombin-clotted plasma containing 125I - labelled DB00142 - plasminogen ( residues 1-790 ) and urokinase . The plasma was kept at 37 degrees C for 0.5 and 10 times the lysis time of the clotted plasma , the clotted plasma until lysis . The plasmin heavy chain from the plasmin-inhibitor complexes was subsequently prepared . Only in one case could a low-grade proteolytic conversion of DB00142 - forms into Lys / DB00134 / DB00161 - forms ( residues 77-790 , 68-790 and 78-790 respectively ) during the preparations be detected . DB00815 / polyacrylamide-gel electrophoresis and N-terminal sequence analysis of the purified plasminogen and plasmin heavy chain showed the following . The plasminogen in plasma was on the DB00142 - form . DB00142 - plasmin constituted 0.74 and 0.58 of the plasmin bound to the alpha 2 - plasmin inhibitor in plasma after brief and prolonged activation respectively . The rest was Lys / DB00134 / DB00161 - plasmin . The clotted plasma contained about equal amounts of DB00142 - plasminogen and Lys / DB00134 / DB00161 - plasminogen , and predominantly Lys / DB00134 / DB00161 - plasmin complexed to alpha 2 - plasmin inhibitor and alpha 2 - macroglobulin . The results of the analysis of the purified material substantiated the identity of radioactive protein bands in the gel after acetic acid / urea / polyacrylamide-gel electrophoresis .

Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 REA ) , epidermal growth factor ( P01133 REA ) and its receptor ( P00533 REA ) , hepatocyte growth factor ( P14210 REA ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 REA ) , vascular endothelial growth factor ( P15692 REA ) , and cyclooxygenase ( P36551 REA ) - 1 and P35354 REA , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 REA , HGFR , P01133 REA , P15692 REA , and P35354 REA , but not P00533 REA , KGF , P21802 REA , P09038 REA , and P23219 REA , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 REA , HGFR , P01133 REA , P15692 REA , and , P35354 REA are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE .

Roles of urokinase type plasminogen activator in a brain stab wound . DB00013 SUB type plasminogen activator ( uPA ) may influence brain pathophysiology after injury . We studied disruption of the blood-brain barrier ( BBB ) and changes in the vasculature after a brain stab wound in uPA-deficient , uPA receptor-deficient , and PA inhibitor - 1 ( P05121 REA ) deficient mice . The extravasation of immunoglobulin was greater in P05121 REA deficient mice ; less pronounced in uPA-deficient mice ; similar to controls in uPA receptor-deficient mice . Vasculatures in the wound proliferated in P05121 REA deficient mice . Our study shows that uPA affects BBB disruption . PA enhances angiogenesis after brain injury .

DB00013 SUB activates macrophage Q15165 REA gene transcription via the PI3K / ROS / MEK / Q12772 REA signalling cascade mediated by the P09619 REA . AIMS : We have recently shown that urokinase plasminogen activator ( uPA ) increases oxidative stress ( OS ) , cholesterol biosynthesis , and paraoxonase 2 ( Q15165 REA ) expression in macrophages via binding to its receptor , the Q03405 REA . Since Q15165 REA is regulated by both OS and cholesterol content , we hypothesized that uPA elicits a cascade of signal transduction events shared by NADPH oxidase and cholesterol biosynthesis that culminates in Q15165 REA gene expression . Here , we investigated the signalling pathway that leads to the expression of Q15165 REA in macrophages in response to uPA . METHODS AND RESULTS : The increase in macrophage Q15165 REA mRNA levels in response to uPA was shown to depend on Q15165 REA gene promoter activation and mRNA transcription . LDL abolished these effects , suggesting a possible role for a transcription factor involved in cellular cholesterogenesis . Indeed , uPA upregulated Q15165 REA expression in a sterol regulatory binding protein - 2 ( Q12772 REA ) - dependent manner , since blocking Q12772 REA maturation by 4 - ( 2 - aminoethyl ) - benzenesulfonyl fluoride abolished uPA-stimulation of Q15165 REA , whereas inhibition of Q12772 REA catabolism by N-acetyl-leucyl-norleucinal had an opposite effect . The upstream signalling mechanisms include uPA activation of extracellular signal-regulated kinases ( P27361 REA / 2 ) , which was dependent on NADPH oxidase and phosphatidylinositol 3 - kinase activation , and these latter effects were mediated by the tyrosine kinase activity of the platelet-derived growth factor receptor-beta . CONCLUSION : These findings provide a framework linking interactions among cellular signalling pathways associated with reactive oxygen species production , macrophage cholesterol biosynthesis , and cellular Q15165 REA expression in vascular pathophysiology .

Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 MENMAX DB00784 MEN ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 REA inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy .

Prevalence of genetic risk factors for coronary artery disease in Corsica island ( France ) . We have investigated the frequencies of seven markers among 100 unrelated individuals with angiographically documented CAD ( Coronary Artery Disease ) and among 100 unrelated healthy blood donors in the central region of Corsica island ( France ) . The seven polymorphisms analyzed were chosen from six candidate genes involved in ( 1 ) P00797 REA - Angiotensin system : Angiotensin converting enzyme ( P12821 REA I / D ) , ( 2 ) Lipid metabolism : DB04540 Ester Transfer Protein gene ( P11597 REA TAQ 1B ) , ( 3 ) Platelet aggregation : alpha and beta subunits of the platelet GpIIb / GpIIIa integrin complex ( GpIIb HPA 3 and GpIIIa Pl ( A1 / A2 ) ) , ( 4 ) Coagulation fibrinolysis : P00747 REA Activator Tissue ( P00750 REA TPA 25 I / D ) and Methylenetetrahydrofolate Reductase ( P42898 REA C677T and A1298C ) . The samples were genotyped using the polymerase chain reaction followed by restriction enzyme analysis for the RFLPs . No significant difference in allele frequencies between patient and control groups was observed . The occurrence of the P42898 REA T677T genotype and of the T677T / A1298A compound genotype is higher in cases ( 20 % ) than in the controls ( 4 % ) . Odds ratio seems to indicate that individuals with the P42898 REA T677T genotype and the T677T / A1298A compound genotype had a 6 - fold increased risk for developing CAD ( ORs = 6 ; 95 % CIs = 1.96- 18.28 ) suggesting a possible association of P42898 REA C677T with the risk of CAD in Corsican population .

DB00013 SUB and its receptors in chronic kidney disease . This review focuses on the role of the serine protease urokinase-type plasminogen activator and its high affinity receptor Q03405 REA / CD87 in chronic kidney disease ( CKD ) progression . An emerging theme is their organ - and site-specific effects . In addition to tubules , uPA is produced by macrophages and fibroblasts in CKD . By activating hepatocyte growth factor and degrading fibrinogen uPA may have anti-fibrotic effects . However renal fibrosis was similar between uPA wild-type and knockout mice in experimental CKD . The Q03405 REA is expressed by renal parenchymal cells and inflammatory cells in a variety of kidney diseases . Such expression appears anti-fibrotic based on studies in Q03405 REA - deficient mice . In CKD Q03405 REA expression is associated with higher uPA activity but its most important effect appears to be due to effects on cell recruitment and migration that involve interactions with a variety of co-receptors and chemoattractant effects of soluble Q03405 REA . P04004 REA and high molecular weight kininogen are alternate Q03405 REA ligands , and receptors in addition to Q03405 REA may also bind directly to uPA and activate cell signaling pathways .

Topical glucocorticoids downregulate P23219 REA positive cells in nasal polyps . BACKGROUND : Influx of inflammatory cells is one of the hallmarks of nasal polyposis . As glucocorticoids ( GC ) are known to exhibit strong anti-inflammatory effects , these drugs are frequently used in the treatment of the disease . Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis . As cyclooxygenases ( P36551 REA ) are key enzymes in the synthesis of both pro - ( P23219 REA , P35354 REA ) and anti-inflammatory prostanoids ( P35354 REA ) , we investigated the role of topical GC on P23219 REA , P35354 REA and inflammatory markers in nasal polyps ( NP ) . METHODS : Immunohistochemical analysis of inflammatory markers ( P34810 REA , CD117 , MBP , elastase , IgE , BB - 1 , P05112 REA , P05113 REA and P05231 REA ) , P23219 REA and P35354 REA was performed on normal nasal mucosa ( NM ) ( n = 18 ) , non-GC treated NP ( n = 27 ) and topical GC treated NP ( n = 12 ) . NP groups were matched for allergy , asthma and ASA intolerance . RESULTS : Increased numbers of eosinophils , P05113 REA + cells and IgE + cells and decreased numbers of mastcells are striking features of NP inflammation ( P < 0.05 ) . In addition , increased numbers of P23219 REA + cells are observed in NP epithelium compared to NM ( P < 0.05 ) . CONCLUSION : Topical GC significantly reduce the number of P23219 REA + NP cells ( P < 0.05 ) , but have no significant effect on P35354 REA + NP cells . No significant reduction in the number of eosinophils is observed for GC treated NP . The number of P05113 REA + cells is however increased significantly upon GC treatment ( P < 0.05 ) .

P14780 REA and urokinase plasminogen activator mediate interleukin - 1 - induced neurotoxicity . Matrix metalloproteinases ( MMPs ) are endopeptidases known to mediate acute neuronal injury , but it is unclear whether these proteases are induced by the primary insult or by inflammation associated with injury . We have reported recently that interleukin - 1 ( IL - 1 ) induces neurotoxicity by an astrocyte-dependent mechanism . The aim of the present study was to test the hypothesis that MMPs mediate IL - 1 neurotoxicity in rat , glial-neuronal cocultures . IL - 1beta induced the release of astrocytic P14780 REA in cocultures , whilst an antagonist of P14780 REA inhibited IL - 1beta - induced neuronal death . DB00013 SUB plasminogen activator ( uPA ) was constitutively expressed on neuronal membrane fractions , and amiloride ( an antagonist of uPA ) or plasminogen activator inhibitor ( P05121 REA ) - 1 significantly reduced IL - 1beta - induced neurotoxicity . Thus , neuronal uPA contributes to IL - 1 neurotoxicity , and may be responsible for activating P14780 REA released from IL - 1 - primed astrocytes . In summary , IL - 1 - induced neurotoxicity is dependent on extracellular protease activity , and these mechanisms may contribute to neuronal cell death in CNS diseases .

DB00013 SUB upregulates matrix metalloproteinase - 9 expression in THP - 1 monocytes via gene transcription and protein synthesis . P00749 REA ( uPA ) is suggested to exert its proliferatory , migratory and invasive action through binding with its membrane receptor , promoting pericellular proteolysis and mediating cell signal transduction . One of the possible actions of urokinase can be related to the regulation of activity and / or the expression of proteolytic enzymes participating in extracellular matrix degradation . In the present study , the role of uPA in regulating matrix metalloproteinase ( MMP ) expression and release by the monocyte cell line THP - 1 was investigated . Recombinant uPA induced the release of P14780 REA / gelatinase B , as detected by zymography and Western blotting , and this release was abolished by actinomycin D and cycloheximide ( inhibitors of DNA transcription and protein synthesis ) and partially suppressed by monensin ( an inhibitor of secretion ) . Proteolytically inactive urokinase with substitution of DB00117 ( 204 ) for Gln was able to reproduce about 70 % of the effect induced by the wild-type recombinant uPA . The reverse transcription-PCR and Northern blot data indicated that the action of r-uPA on THP - 1 cells resulted in formation of P14780 REA mRNA , which depended on time , within 6-48 h , of the cell incubation with r-uPA . These results suggest that urokinase upregulates P14780 REA expression in monocytes via P14780 REA gene transcription and protein biosynthesis .

[ The effect of blood pressure-reducing therapy with captopril on tubular marker excretion in type - 1 diabetics with nephropathy ] . A prospective open clinical trial was carried out with 23 hypertensive type I diabetics ( 13 men , ten women , mean age 49 + / - 9.1 years , duration of diabetes 18 + / - 9.1 years ) with early nephropathy . Glomerular and tubular renal function and metabolic parameters were monitored during 8 months ' treatment with the angiotensin converting enzyme ( P12821 REA ) inhibitor , captopril , in addition to previous antihypertensive treatment with one or more drugs . Blood pressure control tended to improve on captopril ( systolic pressures 152 + / - 13 vs 140 + / - 13 mm Hg , P < 0.05 ; diastolic pressures 89 + / - 10 vs 87 + / - 10 mm Hg , not significant ) . Proteinuria ( > 0.5 g / 24 hours ) fell into the microalbuminuria range ( albumin excretion 2-20 mg / mmol creatinine ) in four out of 13 patients , and microalbuminuria disappeared in four out of ten patients . Urinary levels of the brush border enzyme O60502 ( NAG ) , a marker of tubular dysfunction , were initially raised and fell significantly after 8 months ' treatment with captopril ( 20.3 + / - 14.4 vs 8.8 + / - 8.1 U / g creatinine ; P < 0.01 ) . DB01197 MEN did not affect metabolic control ( HbA 1 , total , HDL and LDL cholesterol , triglycerides , apolipoproteins A1 and B ) or the insulin dosage . These results show that long-term treatment with captopril may favourably influence both albumin excretion and NAG activity , a marker of tubular dysfunction , in type I diabetics with nephropathy .

DB00013 SUB plasminogen activator is a central regulator of macrophage three-dimensional invasion , matrix degradation , and adhesion . DB00013 SUB plasminogen activator ( uPA ) and its receptor ( Q03405 REA ) coordinate a plasmin-mediated proteolytic cascade that has been implicated in cell adhesion , cell motility , and matrix breakdown , for example , during inflammation . As part of their function during inflammatory responses , macrophages move through tissues and encounter both two-dimensional ( 2D ) surfaces and more complex three-dimensional ( 3D ) interstitial matrices . Based on approaches employing uPA gene-deficient macrophages , plasminogen supplementation , and neutralization with specific protease inhibitors , it is reported in this study that uPA activity is a central component of the invasion of macrophages through a 3D Matrigel barrier ; it also has a nonredundant role in macrophage-mediated matrix degradation . For murine macrophages , matrix metalloproteinase - 9 activity was found to be required for these uPA-mediated effects . Evidence for a unique role for uPA in the inverse relationship between macrophage adhesion and 2D migration was also noted : macrophage adhesion to vitronectin was enhanced by uPA and blocked by plasminogen activator inhibitor - 1 , the latter approach also able to enhance in turn the 2D migration on this matrix protein . It is therefore proposed that uPA can have a key role in the inflammatory response at several levels as a central regulator of macrophage 3D invasion , matrix remodeling , and adhesion .

Restoration of flow following haemodialysis catheter thrombus . Analysis of rt-PA infusion in tunnelled dialysis catheters . PROBLEM : DB00013 SUB and streptokinase are commonly used thrombolytic agents for obstructed central venous catheters . Although proven to be efficacious , these agents have the potential to induce fibrin breakdown and streptokinase can not be used repeatedly because of its allergenic nature . Published evidence suggests that DB00013 SUB is safe and effective ( > 70 % efficacy for catheter installation ) and that P00750 REA ( rt-PA ) can achieve as much as 98 % success . OBJECTIVE : To describe our experience with and our protocol for the use of rt-PA as an alternative agent for catheter thrombolysis . DESIGN : Investigation of a cohort of haemodialysis patients with tunnelled central venous catheter ( SPCVC ) placed between December 2001 to August 2003 and who developed catheter thrombus ( female , n = 8 : male , n = 12 ) . Each patient was given an infusion of between 1 and 2 mg rt-PA / h for 4 h . The dose was dependent on partial or total line obstruction . The technical success of rt-PA is defined as returning catheter blood flow to > 250 mL / min for a 4 - h period . FINDINGS : Twenty patients required 57 infusions in 38 lumens between 01/01 / 02 to 01/09 / 03 . For completely blocked lines rt-PA was infused at 2 mg / h for 4 h achieving 85 % success rate . For inadequate flow ( < 250 mL / min ) rt-PA was infused at 1 mg / h for 4 h achieving an 88 % success rate . CONCLUSION : Rt-PA administered at 2 mg / h for blocked lines effectively restores haemodialysis catheter patency , and at 1 mg / h for sluggish lines is also effective in restoring blood flow through catheters .

Sp1 mediates constitutive and transforming growth factor beta-inducible expression of urokinase type plasminogen activator receptor gene in human monocyte-like U937 cells . DB00013 SUB type plasminogen activator receptor ( Q03405 REA ) is known to be involved in conversion of plasminogen into plasmin and its expression can be regulated by a variety of biological agents including transforming growth factor beta ( TGF-beta ) . In the present study , we cloned the promoter region of the human Q03405 REA ( huPAR ) gene ( - 653 to + 61 ) and investigated the transcription regulatory mechanism of the expression of the huPAR gene upon treatment with TGF-beta in human monocyte-like U937 cells . By deletion and point mutational analysis of the huPAR gene promoter , it was found that the sequence positioned at - 70 is required for both constitutive and TGF-beta-inducible expression of the huPAR gene in U937 cells . Using electrophoretic mobility shift assay , we could observe that Sp1 formed a DNA-protein complex at the - 70 sequence . In addition , antisense oligonucleotide against human Sp1 blocked both constitutive and TGF-beta-inducible expression of the luciferase reporter gene driven by the huPAR gene promoter in U937 cells . These results led us to conclude that Sp1 transcription factor mediates constitutive and TGF-beta-inducible expression of the huPAR gene in U937 cells through binding to the sequence located at - 70 .

Association between urokinase-plasminogen activator gene T4065C polymorphism and risk of mitral valve prolapse . BACKGROUND : Abnormalities of collagen and elastic fibers were found in floppy mitral valves ( FMV ) . DB00013 SUB - plasminogen activator ( P00749 REA ) was suggested to be involved in the pathogenesis of elastin and collagen degradation in arterial aneurysm . The role of P00749 REA genetic variant in mitral valve prolapse ( Q14764 REA ) has not been studied . We , therefore , performed a case-controlled study investigating the possible relation between the P00749 REA gene polymorphisms and risk of Q14764 REA in Taiwan Chinese . METHODS : We studied 100 patients with Q14764 REA diagnosed by echocardiography and 106 age - and sex-matched normal control subjects . The T4065C and T3995C polymorphisms of the P00749 REA gene were identified by polymerase chain reaction ( PCR ) - based restriction analysis . RESULTS : There was a significant difference in either the genotype distribution or allelic frequencies between Q14764 REA cases and controls for P00749 REA gene T4065C polymorphism ( P = 0.0001 and 0.0002 , respectively ) . An odds ratio for risk of Q14764 REA associated with P00749 REA T4065C TC genotype was 6.03 ( 95 % confidence interval 2.11- 14.83 ) . An odds ratio for risk of Q14764 REA associated with P00749 REA T4065C T allele was 4.99 ( 95 % confidence interval 1.93- 12.91 ) . There was no significant difference in either the genotype distribution or allelic frequencies between Q14764 REA cases and controls for P00749 REA T3995C polymorphism . Further categorization of the Q14764 REA patients into mild and severe subgroups revealed no statistical difference between these two subgroups for P00749 REA T4065C and T3995C polymorphisms . CONCLUSIONS : This study shows that patients with Q14764 REA have a higher frequency of P00749 REA T4065C TC genotype and T allele that supports a role of the P00749 REA T4065C polymorphism in determining the risk of Q14764 REA among the Chinese population in Taiwan .

Elevated expression of urokinase-like plasminogen activator and plasminogen activator inhibitor type 1 during the vascular remodeling associated with pulmonary thromboembolism . Information is lacking on the mechanisms involved in the organization , resolution , and repair of the vascular lumen after acute pulmonary thromboembolism . Because recent data suggest that the balance between plasminogen activators ( PAs ) and type 1 plasminogen activator inhibitor ( P05121 REA ) plays a role in regulating cell migration within the extracellular matrix , we investigated the expression of these molecules by immunohistochemical and in situ hybridization analysis of pulmonary artery specimens from patients suffering fatal pulmonary embolism . The data were compared with the expression of these molecules in both patients ' noninvolved pulmonary arteries and organ donor pulmonary arteries . Regions of initial organization and vascular remodeling were identified by a modified trichrome stain and by the presence of proliferating cell nuclear antigen ( P12004 REA ) , a cell marker of proliferation . Staining for tissue-type PA antigen was low to undetectable in endothelial cells directly in contact with the fibrin-platelet thromboembolus and in areas in which the endothelial cell lining was replaced by cell growth into the thrombus . DB00013 SUB - like PA ( u-PA ) expression was detected in mononuclear cells within the thrombus in the initial phase of thromboembolism and within cells migrating into the thrombus during the later stages of organization . P05121 REA expression was elevated in the monolayer of endothelial cells underlying the fresh platelet-fibrin thromboembolus and in a P12004 REA - positive cell population present between the pulmonary arterial intima and the thromboembolus that represents early organization . Increased expression of P05121 REA may play a role in inhibiting proteolysis and fostering the localization of the acute fibrin-platelet thrombus to the vascular wall , which is followed by the upregulation of u-PA in migrating cells during the reorganization process .