interacts_with DB08860?
Extracellular signal-regulated kinase and the small GTP-binding protein , Rac , contribute to the effects of transforming growth factor-beta1 on gene expression . The kinases and regulatory proteins that convey signals initiated by transforming growth factor-beta ( TGF-beta ) to the nucleus are poorly characterized . To study the role of the extracellular signal-regulated kinase ( P29323 ) pathway in this process , we transiently transfected NIH 3T3 fibroblasts with TGF-beta-responsive luciferase reporter genes and expression vectors designed to interrupt this kinase cascade . Mitogen-activated protein ( Q96HU1 ) kinase phosphatase-1 and a dominant negative Q96HU1 / P29323 kinase 1 mutant reduced stimulation of plasminogen activator inhibitor-1 ( P05121 ) promoter activity by TGF-beta1 from 11.5- to 4-fold and 4.9-fold , respectively . Similar results were observed with the type I collagen promoters . TGF-beta1 increased P27361 activity 4.5-fold at 5 min and 3 . 1-fold at 3 h , while Jun kinase and p38 activity were not affected . Cotransfection of a dominant negative mutant of the small G protein , Rac , but not dominant negative Ras , Cdc42 , or Rho mutants , reduced the effects of TGF-beta1 on the P05121 promoter by approximately half . In support of a role for Rac in signaling by TGF-beta , GTP binding to Rac was increased 3.7-fold following exposure of NIH 3T3 cells to TGF-beta1 for 3 min . These findings indicate that TGF-beta1 modulates gene expression partly through P29323 and Rac in NIH 3T3 cells . Moving beyond chemotherapy : novel cytostatic agents for malignant mesothelioma . It is now known that vascular endothelial growth factor ( P15692 ) and platelet derived growth factor ( PDGF ) are autocrine growth factors in malignant mesothelioma ; epidermal growth factor receptor ( P00533 ) is also highly overexpressed . Cytotoxic drugs that target these growth factors offer fresh potential for the treatment of mesothelioma . Clinical trials have recently been initiated to evaluate the anti-tumour activity of the P15692 inhibitors SU5416 , bevacizumab and thalidomide . ZD1839 ( DB00317 , AstraZeneca ) , an inhibitor of P00533 tyrosine kinase , is also being evaluated . Two clinical trials are planned to evaluate the two PDGF inhibitors Gleevec ( Imatinib mesylate , STI-571 , Novartis Pharmaceuticals ) and PTK787 ( Novartis Pharmaceuticals ) . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Response of chondrocytes to shear stress : antagonistic effects of the binding partners O00206 and caveolin-1 . Osteoarthritis ( OA ) is often a consequence of excessive mechanical loading of cartilage , which produces hydrostatic stress , tensile strain , and fluid flow . Application of high fluid shear to chondrocytes recapitulates the earmarks of OA , as evidenced by the induction of cyclooxygenase-2 , prostaglandins ( PGs ) , and interleukin-6 ( P05231 ) . Here , we delineated the signaling pathway by which high fluid shear mediates the temporal regulation of P05231 synthesis in human chondrocytes . We determined that O00206 ( O00206 ) and caveolin-1 are binding partners in chondrocytes . Their expression is temporally regulated by fluid shear via the sequential up-regulation of microsomal PGE synthase-1 ( mPGES-1 ) and L-PGDS . High shear stress rapidly induces an 8-fold up-regulation of O00206 expression via an mPGES-1-dependent pathway , whereas prolonged shear exposure concurrently down-regulates O00206 by > 4-fold and up-regulates caveolin-1 expression by > 2.5-fold in an L-PGDS-dependent manner . O00206 and caveolin-1 exert opposing effects on the activation of P27361 /2 , P19957 -K and PKA signaling pathways , which , in turn , regulate the NF-κB-dependent P05231 synthesis in a time-dependent fashion . Reconstructing the signaling network regulating shear-induced P05231 expression in chondrocytes may provide insights for developing therapeutic strategies to combat osteoarthritis . P15692 enhance cortical newborn neurons and their neurite development in adult rat brain after cerebral ischemia . To study the effect of P15692 overexpression on development of cortical newborn neurons in the brains after stroke , we injected human P15692 (165)-expressive plasmids ( phVEGF ) into the lateral ventricle of rat brains with a transient middle cerebral artery occlusion ( MCAO ) . An injection of phVEGF significantly promoted angiogenesis ( BrdU(+)-von Willebrand 's factor(+) ) and reduced infarct volume in the rat brain after MCAO . Single labeling of 5'-bromodeoxyuridine ( BrdU ) and double staining of BrdU with lineage-specific neuronal markers were used to indicate the proliferated cells and maturation of newborn neurons in the brain section of rats at 2 , 4 , and 8 weeks after MCAO . The results showed that BrdU positive ( BrdU(+) ) cells existed in ipsilateral frontal cortex within 8 weeks after MCAO and reached the maximum at 2 weeks of reperfusion . The phVEGF treatment significantly increased BrdU(+) cells compared with the control plasmid ( pEGFP ) injection . Cortical neurogenesis was indicated by the presence of newborn immature ( BrdU(+)-Tuj1(+) ) , newborn mature ( BrdU(+)- P11137 (+) ) , and newborn GABAergic ( BrdU(+)- Q99259 (+) ) neurons . All these neurons declined within 8 weeks after MCAO in the controls . Injection of phVEGF significantly increased BrdU(+)-Tuj1(+) neurons at 2 weeks , and BrdU(+)- P11137 (+) neurons and BrdU(+)- Q99259 (+) neurons at 4 and 8 weeks , respectively after MCAO . Moreover , phVEGF treatment significantly increased neurite length and branch numbers of BrdU(+)- P11137 (+) newborn neurons compared with pEGFP treatment . These results demonstrate that P15692 enhances maturation of stroke-induced cortical neurogenesis and dendritic formation of newborn neurons in adult mammalian brains . Vascular changes after cardiac surgery : role of NOS , P36551 , kinases , and growth factors . Cardiovascular disease remains the leading cause of mortality in the industrialized world . Despite advances in pharmacotherapy and catheter based interventions , coronary artery bypass grafting remains an essential therapeutic modality . The majority of coronary artery bypass operations , as well as other cardiac surgical procedures require the use of ischemic cardioplegic arrest and cardiopulmonary bypass , both of which result in iatrogenic injury to the vasculature and microcirculation . This injury can manifest as impaired vasorelaxation or vasoconstriction , depending upon the organ system involved , resulting in impaired tissue perfusion and the development of edema . Key to this dysfunction are changes in the following : nitric oxide signaling secondary to changes in P29474 and P35228 expression and activity , cyclooxygenase function with increases in pro-inflammatory P35354 activity , alterations in Protein Kinase C and Mitogen Activated Protein Kinase signaling , and an increase in Vascular Endothelial Growth Factor expression increasing vascular permeability and dilatation . This review discusses our current understanding of cardioplegia and cardiopulmonary bypass induced changes in the vasculature , and therapeutic interventions aimed at modulating the altered signaling pathways . The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy . Concomitant expression of urokinase type plasminogen activator ( uPA ) and its surface receptor ( Q03405 ) has been shown to correlate strongly with a more invasive tumor cell phenotype . A highly malignant human epidermoid carcinoma cell line ( HEp3 ) was transfected with a vector capable of expressing an antisense transcript complementary to 300 bases of the 5' end of Q03405 , including the ATG codon . Six stably transfected antisense ( AS-2 , 3 , 5 , 9 , 10 , 12 ) and eight control clones were characterized . All clones produced high levels of uPA activity . Examination of collagenase production and doubling time showed that all of the clones tested produced similar activities . The antisense clones showed a 20-74 % reduction in the Q03405 sites ; the Q03405 mRNA level was also reduced . A test of the invasive ability of all clones in a modified chorioallantoic membrane ( P62158 ) showed that invasiveness of the antisense-inhibited clones was directly proportional to the density of surface Q03405 . The AS-2 clone , which expressed the lowest number of uPARs showed a significantly reduced level of invasion . The invasiveness of additional AS-inhibited clones was also reduced . Seven control and four AS-inhibited clones were tested for tumorigenicity on CAMs of chick embryos . Inoculation of control cells produced large tumors , while the As clones were non-tumorigenic . AS-2 did not produce tumors even if kept in vivo for up to 10 weeks. ( ABSTRACT TRUNCATED AT 250 WORDS ) HMG DB01992 reductase inhibitor suppresses the expression of tissue factor and plasminogen activator inhibitor-1 induced by angiotensin II in cultured rat aortic endothelial cells . BACKGROUND : It has been demonstrated that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors ( HRIs ) reduce the incidence of acute cardiovascular events in patients with hyperlipidemia . Recent reports have shown that the protective effects of these drugs against cardiovascular events are also observed in patients without hyperlipidemia , but the mechanism of this favorable effect still remains unclear . In this study , the effects of HRIs on the endothelial regulation of thrombus formation were elucidated . METHODS AND RESULTS : The mRNA and protein expression of tissue factor ( TF ) and plasminogen activator inhibitor-1 ( P05121 ) induced by angiotensin II ( Ang II ) were evaluated in cultured rat aortic endothelial cells . Pretreatment with simvastatin ( 0.03-3 microg/ml ) significantly inhibited TF and P05121 induction by Ang II in a dose- and time-dependent manner . These inhibitions were significantly attenuated by mevalonic acid or geranylgeranyl pyrophosphate . Both Rho inhibitor , P01024 exoenzyme , and Rho kinase inhibitor , Y-27632 , mimicked the inhibitory effects of simvastatin against TF and P05121 induced by Ang II . This result suggested that the Rho/Rho kinase pathway is related to the TF and P05121 induction by Ang II . CONCLUSION : It was indicated that simvastatin maintains endothelial cells to be antithrombotic by inhibiting TF and P05121 expression via the Rho/Rho kinase pathways in which AngII induces TF and P05121 expression . These observations explain , at least partly , the mechanism of the favorable effects of simvastatin in reducing the thrombotic events . DB08860 inactivates NF-kappaB and decreases P05231 production through Rho kinase pathway in MCF-7 cells . The aim of the present study was to provide new mechanistic insight into the effect of pitavastatin at low dose on NF-kappaB activated by P01375 in the human breast cancer cell line ( MCF-7 ) . We found that treatment of MCF-7 with 1 microM pitavastatin inhibited the proliferation and suppressed the nuclear expression of NF-kappaB p65 induced by P01375 with Western blotting . Furthermore , EMSA showed that pitavastatin significantly reduced the DNA binding activity of NF-kappaB induced by P01375 . Subsequently , luciferase assay revealed that pitavastatin ( 1 microM ) inhibited the transcriptional activity of the NF-kappaB promoter , which was clearly related to the P04035 activity because addition of mevalonic acid ( MEV ) could elevate the NF-kappaB activity . Moreover , the Rho kinase inhibitor Y27632 abolished the effect of pitavastatin on NF-kappaB activity . Finally , the addition of P01375 significantly increased P05231 protein production , which was suppressed by the addition of pitavastatin . These results suggest that pitavastatin at low dose ( 1 microM ) inhibits NF-kappaB activation and decreases P05231 production induced by P01375 . It is dependent on Rho kinase pathway in human breast cancer cells . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . Cystic fibrosis modifier genes related to Pseudomonas aeruginosa infection . Cystic fibrosis ( CF ) is one of the most common life-shortening genetic disorders , and the CF transmembrane conductance regulator ( P13569 ) is the major causal gene . However , a substantial clinical variability among patients with identical P13569 genotypes suggests the presence of modifier genes . We tested the effect of four genes involved in Pseudomonas aeruginosa infection . Analysis of a primary cohort detected eight candidate polymorphisms that were genotyped in the secondary cohort of 1579 patients ; lung function and age at first infection with P. aeruginosa were considered as the phenotypes . Both additive and codominant models were considered , adjusting for confounding variables but not for multiple comparisons . In the secondary cohort , heme oxygenase-1 ( P09601 ) rs2071749 had the most significant effect on lung function in the pediatric group ( P=0.01 ; P(corrected)=0.03 ) , and complement factor 3 ( P01024 ) rs11569393 and P09601 rs2071746 in the adult groups ( P=0.03 for both variants ; P(corrected)=0.16 , 0.09 ) . No polymorphism of complement factor B ( P00751 ) or toll-like receptor 4 ( O00206 ) had a significant modifying effect on lung function in either group . We have identified two genes that showed nominal association with disease severity among CF patients . However , because of the multiple comparisons made , further studies are required to confirm the interaction between these modifying genes and P13569 . DB08860 attenuates the PDGF-induced Q92673 /uPA receptor-mediated migration of smooth muscle cells . Statins , inhibitors of P04035 , elicit various actions on vascular cells including the modulation of proliferation and migration of smooth muscle cells ( SMCs ) . Here , we have elucidated the mechanism by which statins , in particular pitavastatin , attenuate the migration activity of SMCs . The expression of Q92673 , a member of the P01130 family and an enhancer of cell surface localization of urokinase-type plasminogen activator receptor ( Q03405 ) , is increased in cultured SMCs by treatment with DB00102 . DB08860 attenuates the DB00102 -induced surface expression of Q92673 and Q03405 . The increased migration of SMCs observed both upon overexpression of Q92673 and via stimulation of secretion of soluble Q92673 is not reversed by pitavastatin . In vivo studies showed that the SMCs expressing Q92673 in plaques are almost congruent with intimal cells expressing nonmuscle myosin heavy chain ( SMemb ) . DB08860 reduced the expression of Q92673 and SMemb , and the levels of Q92673 , Q03405 , and SMemb in cultured intimal SMCs were reduced to those seen in medial SMCs . We propose that this statin reduces PDGF-induced migration through the attenuation of the Q92673 / Q03405 system in SMCs . Modulation of the Q92673 / Q03405 system with statins suggests a novel treatment strategy for atherogenesis based on suppression of intimal SMC migration . The Kv2.1 K+ channel targets to the axon initial segment of hippocampal and cortical neurons in culture and in situ . BACKGROUND : The Kv2.1 delayed-rectifier K+ channel regulates membrane excitability in hippocampal neurons where it targets to dynamic cell surface clusters on the soma and proximal dendrites . In the past , Kv2.1 has been assumed to be absent from the axon initial segment . RESULTS : Transfected and endogenous Kv2.1 is now demonstrated to preferentially accumulate within the axon initial segment ( AIS ) over other neurite processes ; 87 % of 14 DIV hippocampal neurons show endogenous channel concentrated at the AIS relative to the soma and proximal dendrites . In contrast to the localization observed in pyramidal cells , Q99259 positive inhibitory neurons within the hippocampal cultures did not show AIS targeting . Photoactivable-GFP-Kv2.1-containing clusters at the AIS were stable , moving < 1 microm/hr with no channel turnover . Photobleach studies indicated individual channels within the cluster perimeter were highly mobile ( P42345 tau=10.4+/-4.8 sec ) , supporting our model that Kv2.1 clusters are formed by the retention of mobile channels behind a diffusion-limiting perimeter . Demonstrating that the AIS targeting is not a tissue culture artifact , Kv2.1 was found in axon initial segments within both the adult rat hippocampal P00915 , P00918 , and P07451 layers and cortex . CONCLUSION : In summary , Kv2.1 is associated with the axon initial segment both in vitro and in vivo where it may modulate action potential frequency and back propagation . Since transfected Kv2.1 initially localizes to the AIS before appearing on the soma , it is likely multiple mechanisms regulate Kv2.1 trafficking to the cell surface . Anti-atherogenic and anti-angiogenic activities of polyphenols from propolis . Propolis is a polyphenol-rich resinous substance extensively used to improve health and prevent diseases . The effects of polyphenols from different sources of propolis on atherosclerotic lesions and inflammatory and angiogenic factors were investigated in P01130 gene ( LDLr-/- ) knockout mice . The animals received a cholesterol-enriched diet to induce the initial atherosclerotic lesions ( IALs ) or advanced atherosclerotic lesions ( AALs ) . The IAL or AAL animals were divided into three groups , each receiving polyphenols from either the green , red or brown propolis ( 250 mg/kg per day ) by gavage . After 4 weeks of polyphenol treatment , the animals were sacrificed and their blood was collected for lipid profile analysis . The atheromatous lesions at the aortic root were also analyzed for gene expression of inflammatory and angiogenic factors by quantitative real-time polymerase chain reaction and immunohistochemistry . All three polyphenol extracts improved the lipid profile and decreased the atherosclerotic lesion area in IAL animals . However , only polyphenols from the red propolis induced favorable changes in the lipid profiles and reduced the lesion areas in AAL mice . In IAL groups , VCAM , P13500 , FGF , PDGF , P15692 , PECAM and P14780 gene expression was down-regulated , while the metalloproteinase inhibitor P01033 gene was up-regulated by all polyphenol extracts . In contrast , for advanced lesions , only the polyphenols from red propolis induced the down-regulation of P16671 and the up-regulation of P09601 and P01033 when compared to polyphenols from the other two types of propolis . In conclusion , polyphenols from propolis , particularly red propolis , are able to reduce atherosclerotic lesions through mechanisms including the modulation of inflammatory and angiogenic factors . A review and proposed nomenclature for major proteins of the milk-fat globule membrane . The characteristics and possible functions of the most abundant proteins associated with the bovine milk-fat globule membrane are reviewed . Under the auspices of the Milk Protein Nomenclature Committee of the ADSA , a revised nomenclature for the major membrane proteins is proposed and discussed in relation to earlier schemes . We recommend that proteins be assigned specific names as they are identified by molecular cloning and sequencing techniques . The practice of identifying proteins according to their Mr , electrophoretic mobility , or staining characteristics should be discontinued , except for uncharacterized proteins . The properties and amino acid sequences of the following proteins are discussed in detail : P15941 , xanthine dehydrogenase/oxidase , P16671 , butyrophilin , adipophilin , periodic acid Schiff 6/7 ( DB00233 6/7 ) , and fatty acid binding protein . In addition , a compilation of less abundant proteins associated with the bovine milk-fat globule membrane is presented . A new P04035 inhibitor , pitavastatin remarkably retards the progression of high cholesterol induced atherosclerosis in rabbits . BACKGROUND : The remarkable anti-atherosclerotic effects of 3-hydroxy-3-methyl-glutaryl- DB01992 reductase inhibitor have not been demonstrated in diet induced severe hyperlipidemia in rabbit model . OBJECTIVE : We have investigated the effect of pitavastatin , a newly developed statin , on atherosclerosis in rabbits . METHODS AND RESULTS : Oophorectomized female NZW rabbits were fed 0.3 % cholesterol chow for 12 weeks with or without pitavastatin ( 0.1mg/kg per day ) ( Gp.NK and HCD ) . The level of serum cholesterol was decreased in Gp.NK compared with Gp.HCD ( 772.8 +/- 70.2 versus 1056.9 +/- 108.3 mg/d ) , whereas no significant alterations were observed in triglyceride and HDL-cholesterol . NO dependent response stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-l-arginine acetate were all improved by pitavastatin treatment . DB08860 treatment increased the level of cyclic GMP in the aorta of cholesterol fed rabbits . In the aorta , the expression of P29474 mRNA was significantly up regulated and O(2)(-) production was slightly reduced in Gp.NK animals . Atherosclerotic area was significantly decreased in aortic arch and thoracic aorta from Gp.NK compared with those from Gp.HCD ( 15.1 +/- 5.3 versus 41.9 +/- 10.2 % , 3.1 +/- 1.1versus 7.9 +/- 1.2 % in Gp.NK and Gp.HCD aortic arch and thoracic aorta ) . Anti-macrophage staining area , the P03956 or 2 and the nitrotyrosine positive area were decreased in Gp.NK . CONCLUSION : DB08860 retards the progression of atherosclerosis formation and it improves NO bioavailability by P29474 up-regulation and decrease of O(2)(-) . MEK/ P29323 -dependent Q03405 expression is required for motility via phosphorylation of P70S6K in human hepatocarcinoma cells . Motility and invasiveness events require specific intracellular signaling cascade activations . In cancer liver cells , one of these mechanisms could involve the MAPK MEK/ P29323 cascade activation which has been shown over expressed and activated in hepatocellular carcinoma . To study whether the MEK/ P29323 cascade is involved in the motility of HCC , we examined the effect of MEK inhibitor and P28482 silencing using monolayer wound-healing assays and fluoroblock invasion systems . Evidence was provided that the MAPK cascade is a key transduction pathway which controls HCC cells motility and invasiveness . We could disconnect proliferation to motility using mitomycin C and we established that RNAi-mediated inhibition of P28482 led to strongly reduced cell motility . To improve our understanding , we analysed the regulation and the role of urokinase receptor ( Q03405 ) in this process . We provided evidence that Q03405 was under a MEK/ P29323 dependent mechanism and blocking Q03405 activity using specific antagonist or inhibiting its expression by RNA interference which resulted in complete inhibition of motility . Moreover , we found in MAPK inhibited cultures and in Q03405 silencing cells that p70S6K phosphorylation on residue DB00156 -389 was significantly reduced , whereas DB00133 -421/ DB00156 -424 phosphorylation did not change . We highlighted that the P42345 / P42345 pathway did not affect motility and DB00156 -389 phosphorylation . Furthermore , we demonstrated that p70S6K inhibition by RNA interference completely inhibited hepatocarcinoma cell motility . Therefore , targeting Q03405 and/or MEK/ P29323 /S6K by RNA interference could be a major therapeutic strategy for the future treatment of invasive hepatocarcinoma cells . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . DB08860 , a potent hydroxymethylglutaryl coenzyme a reductase inhibitor , increases cholesterol 7 alpha-hydroxylase gene expression in HepG2 cells . BACKGROUND : The effect of pitavastatin on the mRNA levels of apolipoprotein ( apo ) A-I , peroxisome proliferator-activated receptor alpha ( PPARalpha ) , cholesterol 7alpha-hydroxylase ( P22680 ) , and farnesoid X receptor ( Q96RI1 ) in HepG2 cells was examined to establish whether pitavastatin affects bile acid synthesis and if so , to determine a possible molecular mechanism . METHODS AND RESULTS : HepG2 cells were cultured in serum-free Dulbecco 's modified Eagle medium for 18 h before drug treatment . Total RNA was extracted at set times and mRNA levels were quantified by reverse transcription-real time polymerase chain reaction . DB08860 at 0.1 , 1 , 5 , and 10 micromol/L increased the mRNA levels of apo A-I , PPARalpha , P22680 , and Q96RI1 in a dose-dependent manner . The mRNA levels of apo A-I , Q07869 alpha , P22680 , and Q96RI1 similarly increased with increasing doses of pitavastatin . Coincubation of mevalonate ( 4 mmol/L ) with pitavastatin ( 5 micromol/L ) reversed the inductive effects of pitavastatin on the mRNA levels of these genes , indicating that the inductive effects of pitavastatin were related to its inhibition of P04035 . CONCLUSIONS : DB08860 increased the mRNA levels of P22680 in HepG2 cells , suggesting that increased conversion of cholesterol to bile acids may be the mechanism for its potent low-density lipoprotein cholesterol-lowering effects . DB08860 , an P04035 inhibitor , ameliorates endothelial function in chronic smokers . BACKGROUND : Smoking is a major cardiovascular risk factor , leading to endothelial dysfunction . The present study investigated the hypothesis that pitavastatin , an P04035 inhibitor , may improve endothelial function in chronic smokers via its antioxidant properties . METHODS AND RESULTS : The 30 male chronic smokers who exhibited mild hypercholesterolemia at the time of physical check-up were enrolled and randomized to the pitavastatin group ( 2 mg/day , n=15 ) or the untreated control group ( n=15 ) . Before and after the 4-week treatment period , endothelium-dependent flow-mediated dilation ( FMD ) and endothelium-independent dilation by glyceryl trinitrate ( GTD ) were examined , and the FMD/GTD ratio was calculated . The pitavastatin group showed a significant restoration of endothelial function ( percent change in FMD : +49.6 % vs +1.4 % ; percent change in FMD/GTD ratio : +26.6 % vs 4.5 % , P < 0.05 respectively ) , and a significant reduction in oxidative stress levels ( malondialdehyde-low-density lipoprotein-cholesterol : 16.6 % vs +7.5 % ; free radical activity : 1.8 % vs +9.7 % , P < 0.05 respectively ) compared with the control group . DB08860 had no effect on the number of circulating P28906 (+)CD133(+) progenitor cells , endothelial progenitor cells , or the P08253 , P14780 and P15692 levels . In vitro oxidative stress monitoring assay revealed that pitavastatin protected endothelial cells against oxidative stress . CONCLUSIONS : DB08860 restores endothelial function , even in chronic smokers , possibly through its antioxidative properties . ( Circ J 2010 ; 74 : 195 - 202 ) . DB08860 inhibits vascular smooth muscle cell proliferation by inactivating extracellular signal-regulated kinases 1/2 . We recently reported that lysophosphatidylcholine ( lysoPC ) acts on vascular smooth muscle cells ( VSMCs ) to produce a mitogenic response through the activation of extracellular signal-regulated kinases 1/2 ( P27361 /2 ) . In this study , we examined the role of P04035 inhibitors on lysoPC-induced VSMC proliferation . DB08860 , a new P04035 inhibitor , suppressed lysoPC-induced DNA synthesis in primary cultured rat VSMCs . Since lysoPC-induced P27361 /2 activation contributes to smooth muscle cell proliferation , we explored the effect of pitavastatin on P27361 /2 activation . DB08860 inhibited lysoPC-induced P27361 /2 phosphorylation and P27361 /2 activation . The other P04035 inhibitors , atrovastatin and fluvastatin , also inhibited lysoPC-induced P27361 /2 phosphorylation . DB08860 also inhibited lysoPC-induced c-fos mRNA expression . To gain insight into the mechanism of the inhibitory effect of pitavastatin on P27361 /2 activation by lysoPC , we examined the role of the mevalonate pathways . Mevalonate and farnesylpyrophosphate reduced the inhibition of P27361 /2 phosphorylation by pitavastatin . These studies demonstrate that pitavastatin may inhibit lysoPC-induced VSMC proliferation , at least in part , by inactivating P27361 /2 , which is linked to mevalonate metabolism . Novel , small molecule induced GABA-hATSCs for targeting of neuropathic pain . Recent study showed that ROS has a crucial function during neuropathic pain development and maintenance . In this study , we suggest that a small , novel molecule , CMB-1078 , can effectively induce GABAergic neuronal differentiation from human adipose tissue-derived stromal cells ( hATSCs ; GABA-hATSCs ) , which play a key role in ameliorating neuropathic pain caused by spinal cord injury . Compared to control hATSCs , the engraftment of GABA-hATSCs into animals with neuropathic pain significantly reduced secondary injury , including inflammation , GABAergic neuronal degeneration , and the circulation or propagation of proinflammatory factors cyclooxygenase2 ( P35354 ) , interlukin-1 β ( IL-1β ) , P04839 ( NOX 2 ) , Q9NPH5 ( NOX 4 ) and tumor necrosis factor α ( TNFα ) into the lesion . At the protein level , we also demonstrated that GABA-hATSCs engrafted into animals with neuropathic pain increased glutamic acid decarboxylase 65 ( Q05329 ) and glutamic acid decarboxylase 67 ( Q99259 ) expression levels . In addition , we evaluated functional pain behavior in the GABA-hATSCs- or control hATSCs-engrafted animal group , the pain in the PBS-infused animal group , and healthy animals by measuring mechanical and heat sensitivity . The pain plus GABA-hATSCs-engrafted animal groups showed paw withdrawal thresholds ( PWTs ) that gradually improved . In contrast , the mice with neuropathic pain did not show improved PWT . Further , the control hATSCs-engrafted animal showed attenuated PWTs . Finally , we suggest that the molecular function of GABA-hATSCs in neuropathic pain may provide potential therapeutic tools for the treatment of pain by controlling the pathology of neuropathic pain through neuroprotection and regeneration . The novel P04035 inhibitor , DB08860 , induces a protective action in vascular endothelial cells through the production of nitric oxide ( NO ) . This study sought to induce the effect of nitric oxide ( NO ) production in vascular endothelial cells by DB08860 , which is a novel P04035 inhibitor ( statin ) . The growth capacity of vascular endothelial cells significantly ( p < 0.01 ) declined when stimulated with P01375 ( 10 ng/ml ) . The growth capacity of the P01375 treated cells recovered , when the P01375 stimulation was performed after DB08860 ( 100 nM ) pretreatment . The recovery of the growth capacity of the cells was suppressed by the presence of the NO synthase inhibitor , L-NAME . DB08860 increased NO production by the vascular endothelial cells in a dose and time dependent manner . The NO production was suppressed by the presence of mevalonic acid and geranylgeranyl pyrophosphate . In addition , the expression of endothelial nitric oxide synthase was strongly induced by DB08860 , and was suppressed by mevalonic acid and geranylgeranyl pyrophosphate by Western blot analysis . Our results show that DB08860 induces NO production by vascular endothelial cells , and protects vascular endothelial cells from injury due to the inflammatory reaction induced by P01375 . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . P10275 coregulator Q96L73 -alpha interacts with death receptor-6 revealed by the yeast two-hybrid . Q96L73 -alpha is a newly identified androgen receptor coactivator . In order to further elucidate its precise role in cells , using the Q96L73 -alpha fragment containing four P20941 and one Q01105 conserved domains as bait we revealed an Q96L73 -alpha- P20941 - Q01105 -interacting protein , death receptor-6 ( O75509 ) , in the yeast two-hybrid screening . O75509 is the member of P01375 receptor family and has a death domain in its intracellular cytoplasmic portion ( DR6cp ) to mediate the cell apoptosis . The interaction between Q96L73 -alpha- P20941 - Q01105 and DR6cp was confirmed in vitro and in vivo . Our finding implied that androgen signaling pathway might cross talk with apoptosis signaling pathway through the interaction between Q96L73 -alpha and O75509 . DB08860 prevents DB01221 -induced retinal ganglion cell death by suppressing leukocyte recruitment . Excitotoxicity is a major cause of retinal ganglion cell ( RGC ) death during ischemic diseases such as vessel occlusion and diabetic retinopathy . However , the underlying mechanisms are not well understood . Statins , inhibitors of the P04035 , have neuroprotective effects in addition to their original role in lowering cholesterol . We hypothesize that pitavastatin , a recently introduced potent statin , is protective against N-methyl-d-aspartic acid ( DB01221 ) -induced RGC death . DB08860 , administered by gavage , abolished DB01221 -induced loss of RGCs . To elucidate the mechanisms underlying the neuroprotective effect of pitavastatin , we investigated its impact on inflammation . DB01221 increased the expression of interleukin-1beta and P01375 , and endothelial adhesion molecules , including P05362 , and induced leukocyte accumulation in the retinal vessels . DB08860 significantly reduced DB01221 -induced leukocyte accumulation and up-regulation of endothelial adhesion molecules , whereas cytokine expression was unaffected . Systemic blockade of P05362 in wild-type mice or absence of P05107 in gene-deficient ( P05107 (-/-) ) mice significantly suppressed DB01221 -induced leukocyte accumulation and RGC death . These findings suggest a novel and causative role for inflammatory leukocyte recruitment in DB01221 -induced excitotoxicity . Furthermore , we show the novel neuroprotective effect of statins against excitotoxicity-induced RGC death . Statins or other anti-inflammatory agents may thus have therapeutic benefits in excitotoxicity-associated neuronal diseases through blockade of leukocyte recruitment . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . DB08860 suppresses acute and chronic rejection in murine cardiac allografts . INTRODUCTION : P04035 inhibitors play several roles in the maintenance of organ transplants . We investigated the role of pitavastatin , a potent and newly developed P04035 inhibitor , in cardiac allograft rejection and mechanism of graft arterial disease ( Q99259 ) suppression . METHODS : Balb/c mice hearts were transplanted into C3H/He mice ( a full allomismatch combination ) to assess acute rejection or C57BL/6 hearts into B6.C-H2 ( < bm12 > ) KhEg ( a class II mismatch combination ) to examine the extent of Q99259 . DB08860 was administered orally to mice everyday ( 3 mg/kg/day ) . To assess the effect in acute rejection , mixed lymphocyte reaction was performed and cytokine mRNA expression was examined with ribonuclease protection assay . RESULTS : DB08860 significantly prolonged allograft survival . Lymphocyte proliferation was inhibited by pitavastatin , and RPA showed down-regulation of interleukin-6 in pitavastatin-treated cardiac allografts . Allografts in the pitavastatin-treated group after 8 weeks showed less Q99259 compared with the control group . In vitro , pitavastatin suppressed the smooth muscle cell proliferation in response to activated T cells and inhibited extracellular signal-regulated kinase 1/2 activation . CONCLUSION : DB08860 could be effective in the suppression of acute rejection and Q99259 development in cardiac transplantation . Anti-adipogenic action of pitavastatin occurs through the coordinate regulation of PPARgamma and Pref-1 expression . BACKGROUND AND PURPOSE : Adipocyte differentiation in vitro is coordinately activated by two transcription factors , peroxisome proliferator-activated receptor gamma ( PPARgamma ) and CCAAT enhancer binding protein alpha ( C/EBPalpha ) , but it is inhibited by preadipocyte factor-1 ( pref-1 ) . Statins , inhibitors of P04035 and de novo cholesterol synthesis , can have pleiotropic effects which influence adipocyte phenotype by ill-defined mechanisms . We investigated the effects of pitavastatin ( NK-104 ) on adipocyte differentiation and the transcriptional pathways involved . EXPERIMENTAL APPROACH : The effects of pitavastatin on adipocyte differentiation were evaluated by the formation of oil droplets , content of cellular triglyceride and expression of adipocyte-specific genes . Regulatory mechanisms were assessed by analysis of PPARgamma , C/EBPalpha and pref-1 expression . KEY RESULTS : DB08860 significantly inhibited adipocyte differentiation of 3T3- Q9NUQ9 preadipocytes in response to adipogenic inducers . Evidence for inhibition included fewer Oil Red O positive droplets , less cellular triglyceride and decreased expression of adipocyte-specific genes , including fatty acid binding protein ( aP2 ) , P16671 , adipsin and glucose transporter 4 ( P14672 ) . The inhibitory effects of pitavastatin on adipocyte differentiation of 3T3- Q9NUQ9 preadipocytes were time and concentration dependent . DB08860 significantly blocked induction of PPARgamma expression , but not C/EBPalpha expression or DNA binding activity of PPARgamma . Also , pitavastatin induced pref-1 expression in preadipocytes and maintained expression of pref-1 at high levels in differentiated cells . CONCLUSIONS AND IMPLICATIONS : Our data suggest that pitavastatin inhibits adipocyte differentiation by blocking PPARgamma expression and activating pref-1 expression . These studies may have implications in the regulation of adipogenesis in response to statins . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . DB08860 up-regulates the induction of P35228 through enhanced stabilization of its mRNA in pro-inflammatory cytokine-stimulated hepatocytes . Studies have indicated that protective effects of statins ( P04035 inhibitor ) are associated with the regulation of endothelial nitric oxide synthase ( P29474 ) or inducible NOS ( P35228 ) in heart and liver diseases . Statins have been reported to enhance hepatic NO production and decrease the vascular tone in patients with cirrhosis . However , it is unclear which NOS contributes to the increased NO production . We hypothesized that statins are involved in the up-regulation of P35228 in inflammatory liver , resulting in decreased hepatic resistance . Primary cultured rat hepatocytes were treated with pro-inflammatory cytokine interleukin ( IL ) -1beta in the presence or absence of pitavastatin . Pretreatment of cells with pitavastatin resulted in up-regulation of P35228 induction by IL-1beta , followed by increased NO production . DB08860 had no effects on the degradation of IkappaB or activation of NF-kappaB . However , pitavastatin super-induced the up-regulation of type I IL-1 receptor ( IL-1RI ) , which is essential for P35228 induction in addition to the IkappaB/NF-kappaB pathway . Mevalonate and geranylgeranylpyrophosphate blocked the stimulatory effects of pitavastatin on P35228 and IL-1RI induction . Transfection experiments revealed that pitavastatin increased the stability of P35228 mRNA rather than its promoter transactivation . In support of this observation , pitavastatin increased the antisense-transcript corresponding to the 3'-UTR of P35228 mRNA , which stabilizes P35228 mRNA by interacting with the 3'-UTR- and RNA-binding proteins . These findings demonstrate that pitavastatin up-regulates P35228 by the stabilization of its mRNA , presumably through the super-induction of IL-1RI and antisense-transcript . This implies that statins may contribute to a novel potentiated treatment in liver injuries including cirrhosis . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . P10275 promotes esophageal cancer cell migration and proliferation via matrix metalloproteinase 2 . Esophageal squamous cell carcinoma ( ESCC ) is one of the most common malignancies worldwide . P10275 ( AR ) plays an important role in many kinds of cancers . However , the molecular mechanisms of AR in ESCC are poorly characterized . In the present study , Western blot analysis and real-time quantitative PCR were performed to identify differentially expressed AR in 40 ESCC tissue samples , which revealed that the messenger RNA ( mRNA ) and protein expression of AR is upregulated in the ESCC tissue samples . AR overexpression induced increases in ESCC cell invasion and proliferation in vitro . Silencing of AR inhibited the proliferation of KYSE450 cells which have a relatively high level of AR , and the invasion of KYSE450 cells was distinctly suppressed . Furthermore , AR knockdown led to substantial reductions in matrix metalloproteinase 2 ( P08253 ) and p-AKT levels in ESCC cell lines , but no significant change in AKT and P14780 expression . These results suggest that AR is involved in tumor progression , and thus , AR could represent selective targets for the molecularly targeted treatments of ESCC . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Lack of endothelial nitric oxide synthase aggravates murine pneumococcal meningitis . DB00435 ( NO ) plays a central role in the pathogenesis of bacterial meningitis . However , the role of NO produced by endothelial NO synthase ( P29474 ) in meningitis is still unclear . We investigated the influence of P29474 depletion on the inflammatory host response , intracranial complications , and outcome in experimental pneumococcal meningitis . Leukocyte accumulation in the cerebrospinal fluid was more pronounced in infected P29474 -deficient mice than in infected wild type mice . This effect could be attributed to an increased expression of P16109 , macrophage inflammatory protein-2 , keratinocyte-derived cytokine , and interleukin ( IL ) -1beta in the brain of infected P29474 -deficient mice . However , no differences in the cerebral expression of intercellular adhesion molecule-1 , tumor necrosis factor-alpha , and P05231 as well as of neuronal NOS and inducible NOS could be detected between infected wild type and mutant mice . In addition to enhanced leukocyte infiltration into the P04141 , meningitis-associated intracranial complications including blood-brain barrier disruption and the rise in intracranial pressure were significantly augmented in infected P29474 -deficient mice . The aggravation of intracranial complications was paralleled by a worsening of the disease , as evidenced by a more pronounced hypothermia , an enhanced weight reduction , and an increased death rate . The current data indicate that P29474 deficiency is detrimental in bacterial meningitis . This effect seems to be related to an increased expression of ( certain ) cytokines/chemokines and adhesion molecules ; thus leading to increased meningeal inflammation and , subsequently , to aggravated intracranial complications . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Novel antiangiogenic pathway of thrombospondin-1 mediated by suppression of the cell cycle . We have recently reported that keratin 14-promoter-driven vascular endothelial growth factor ( P15692 ) -E(NZ-7) transgenic mice have a significant number of capillary vessels in subcutaneous tissue . However , these vessels are generated in a layer some distance from the epithelial basal cells that express Q9NRA1 (NZ-7) , suggesting that one or more antiangiogenenic molecules may exist very near the basal cell layer . By screening keratinocyte-conditioned medium , we found that thrombospondin-1 ( P07996 -1 ) is produced from keratinocytes and suppresses human umbilical vein endothelial cells ( HUVEC ) growth as well as tubular formation in a HUVEC-fibroblast coculture system . Different to the known mechanism of P16671 -dependent endothelial cell apoptosis , the HUVEC we used did not express P16671 at detectable levels , indicating a new mechanism for P07996 -1-induced antiangiogenesis . We found that P07996 -1 induces little apoptosis of endothelial cells but causes cell-cycle arrest , increasing the amounts of P38936 (CIP/WAF-1) and unphosphorylated retinoblastoma ( Rb ) in HUVEC . P16671 -binding peptide in P07996 -1 and P16671 -neutralizing antibody did not block the P07996 -1-induced cell-cycle arrest . Our results strongly suggest that P07996 -1 utilizes a novel pathway for its antiangiogenic effect independent of P16671 , and suppresses the cell cycle . Oxidized LDL and lysophosphatidylcholine stimulate plasminogen activator inhibitor-1 expression through reactive oxygen species generation and P27361 /2 activation in 3T3- Q9NUQ9 adipocytes . P00747 activator inhibitor-1 ( P05121 ) is secreted from adipose tissue and is considered to be a risk factor for both atherosclerosis and insulin resistance . Here we report for the first time that P05121 expression is enhanced by oxidized low-density lipoprotein ( OxLDL ) and its lipid component lysophosphatidylcholine ( Q16549 ) in mouse 3T3- Q9NUQ9 adipocytes . In fully differentiated 3T3- Q9NUQ9 cells , OxLDL treatment increased the mRNA expression and protein secretion of P05121 in a dose- and time-dependent manner , whereas native LDL had no effect . The addition of an anti- P16671 antibody suppressed OxLDL-stimulated P05121 expression by 50 % , suggesting that adipose-derived P16671 contributes to roughly half of the P05121 expression stimulated by OxLDL . In addition , pharmacological experiments showed that the OxLDL-stimulated enhancement in P05121 expression was mediated through the generation of reactive oxygen species ( ROS ) and phosphorylation of extracellular signal-regulated kinase 1/2 . Furthermore , Q16549 , a major lipid component of OxLDL , was responsible for the enhanced expression of P05121 as phospholipase A(2)-treated acetyl LDL , which generates Q16549 , strongly stimulated P05121 expression , whereas acetyl LDL itself had no such activity . These data demonstrate that the uptake of OxLDL and , in particular , its lipid component Q16549 into adipocytes triggers aberrant ROS-mediated P05121 expression , which may be involved in the pathogenesis of metabolic syndrome . Managing the underlying cause of cystic fibrosis : a future role for potentiators and correctors . Cystic fibrosis ( CF ) , a severe genetic disease , is caused by mutations that alter the structure and function of P13569 , a plasma membrane channel permeable to chloride and bicarbonate . Defective anion transport in CF irreversibly damages the lungs , pancreas , liver , and other organs . CF mutations cause loss of P13569 function in multiple ways . In particular , class 3 mutations such as p.Gly551Asp strongly decrease the time spent by P13569 in the open state ( gating defect ) . Instead , class 2 mutations impair the maturation of P13569 protein and its transport from the endoplasmic reticulum to the plasma membrane ( trafficking defect ) . The deletion of phenylalanine 508 ( p.Phe508del ) , the most frequent mutation among CF patients ( 70-90 % ) , destabilizes the P13569 protein , thus causing both a trafficking and a gating defect . These two defects can be overcome with drug-like molecules generically called correctors and potentiators , respectively . The potentiator Kalydeco™ ( also known as DB08820 or VX-770 ) , developed by Vertex Pharmaceuticals , has been recently approved by the US FDA and the European Medicines Agency ( P15941 ) for the treatment of CF patients carrying at least one P13569 allele with the p.Gly551Asp mutation ( 2-5 % of all patients ) . In contrast , the corrector VX-809 , which significantly improves p.Phe508del- P13569 trafficking in vitro , is still under study in clinical trials . Because of multiple defects caused by the p.Phe508del mutation , it is probable that rescue of the mutant protein will require combined treatment with correctors having different mechanisms of action . This review evaluates the status of experimental and clinical research in pharmacotherapy for the CF basic defect . P01375 polymorphisms as a potential modifier gene in the cystic fibrosis . Modifier genes , as the P01375 -α gene , can modulate the cystic fibrosis ( CF ) severity . Thus , -238G > A and -308G > A polymorphisms of P01375 -α gene were analyzed as modifiers of CF . In this context , the present study enrolled 49 CF patients ( diagnosis performed by sweat test and complete P13569 screening ) . The -238G > A polymorphism analysis was performed by Q9ULH0 -PCR , and -308G > A , by PCR-RFLP . In our data , the -238G > A polymorphism was not associated with clinical variability . The AA genotype for -308G > A polymorphism was a risk factor for early gastrointestinal symptoms ( OR=5.98 , 95 % CI=1.06-49.68 ) and protection for the first Pseudomonas aeruginosa ( OR=0.05 , 95 % CI=0.0003-0.007 ) . For the first P. aeruginosa , GA genotype was a risk factor ( OR=10.2 , 95 % CI=1.86-84.09 ) ; for the same genotype , the diagnosis was made in minor time than the AA genotype ( p=0.031 ) . Considering the -308G > A polymorphism alleles , the G allele was a risk factor for early pulmonary symptoms ( OR=3.81 , 95 % CI=1.13-12.97 ) and P. aeruginosa ( OR=66.77 , 95 % CI=15.18-482.7 ) ; however , the same allele showed better transcutaneous oxygen saturation ( OR=9.24 , 95 % CI=1.53-206.1 ) . The A allele was a protective factor for early pulmonary symptoms ( OR=12.26 , 95 % CI=0.08-0.89 ) and P. aeruginosa ( OR=12.15 , 95 % CI=0002-0007 ) , however , the same allele was a risk factor for worst transcutaneous oxygen saturation ( OR=7.01 , 95 % CI=1.14-157.4 ) . As conclusion , the -308G > A polymorphism of the P01375 -α gene was associated with the CF severity . P10244 prevents growth arrest associated with terminal differentiation of monocytic cells . P10244 is a transcriptional regulator of gene expression and is highly homologous to c-Myb in its N-terminal DNA binding domain . However , unlike c-myb , whose expression is restricted largely to immature hematopoietic cells , B-myb mRNA has been found to be expressed in all proliferating mammalian cell lines and is clearly regulated in a cell cycle dependent manner . That c-Myb and P10244 proteins perform different roles in proliferation and/or differentiation is suggested by the redundancy of their expression . It was previously shown that degenerated c-Myb expression can inhibit P05231 induced terminal differentiation of the leukemia cell line M1 . We found that , unlike the downregulation of c-Myb protein which is an early response of progenitor M1 cells to P05231 treatment , the downregulation of P10244 occurs late , just prior to terminal differentiation and growth arrest . It was , therefore , of interest to examine the role of the murine P10244 protein in the proliferation and differentiation of the M1 cells and to compare these effects to those of c-Myb in the same system . Clones ectopically producing P10244 , like those ectopically expressing c-Myb , proliferated in the presence of the differentiation-inducing agent and did not undergo the programmed cell death which normally follows terminal macrophage differentiation . In addition , the cell-cycle distribution of M1/ P10244 cells was comparable to untreated cells . Although M1/ P10244 and M1/c-Myb clones treated with P05231 appeared quite immature , differentiation markers were demonstrated to be maintained at near normal levels ( e.g. MyD88 , Mac-2 ) , or be partially reduced in expression ( P01024 , Fc and Mac-1 receptors ) suggesting that the cells had undergone commitment to maturation , but were unable to terminally differentiate . 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 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , 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 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Candidate genetic markers and the risk of restenosis after coronary angioplasty . The aim of the present study was to test for possible associations between candidate gene polymorphisms and the risk of restenosis and recurrent restenosis after percutaneous transluminal coronary angioplasty ( PTCA ) without stenting . We followed up 511 PTCA patients , and restenosis and recurrent restenosis were defined according to angiographical criteria . Genotyping of the beta-fibrinogen -455 G/A , glycoprotein ( GP ) IIIa PlA1/PlA2 , plasminogen activator inhibitor-1 ( P05121 ) 4G/5G , factor V Leiden 1691 G/A , tumour necrosis factor alpha ( TNFalpha ) -238 G/A , TNFalpha -308 G/A , interleukin ( IL ) -1alpha -889 C/T , IL-1beta -511 C/T , methylenetetrahydrofolate reductase ( P42898 ) 677 C/T and endothelial nitric oxide synthase ( P29474 ) 4 b/a gene polymorphisms was performed by PCR and restriction-fragment-length-polymorphism-based techniques . One hundred and sixty patients ( 31.3 % ) developed restenosis and in 130 of these patients , of whom 123 were available for analysis , a second PTCA without stenting was performed . Of these patients , 35 ( 28.5 % ) developed recurrent restenosis . None of the investigated genotypes were associated with the risk of restenosis or recurrent restenosis after PTCA . The degree of stenosis before and immediately after PTCA and the severity of the lesion were independent predictors for restenosis after PTCA . In conclusion , there was no association between the beta-fibrinogen -455 G/A , GP IIIa PlA1/A2 , P05121 4G/5G , factor V Leiden 1691 G/A , TNFalpha -238 G/A , TNFalpha -308 G/A , IL-1alpha -889 C/T , the IL-1beta -511 C/T , P42898 677 C/T and P29474 4 b/a gene polymorphisms and the risk of restenosis after PTCA as well as recurrent restenosis after repeated PTCA . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . Genes controlling spread of breast cancer to lung " gang of 4 " . Cancer-related mortality is caused in a large part by the metastasis of primary tumor . Each cancer has a particular way of spreading cancerous cells . Recently , genetic and pharmacological analysis identified the set of genes , such as epidermal growth factor receptor ligand epiregulin ( O14944 ) , cyclooxygenase-2 ( P35354 ) and matrix metalloproteinases 1 and 2 ( P03956 and P08253 ) that have been found to be associated with metastasis of breast cancer to lung . Inhibition of P00533 and P35354 could minimize lung metastasis of breast cancer in a clinical setting . In this review , we summarized the current knowledge on O14944 , P35354 , P03956 and P08253 in tumor development and metastasis . Induction of autophagy is an early response to gefitinib and a potential therapeutic target in breast cancer . Gefitinib ( DB00317 (®) , ZD1839 ) is a small molecule inhibitor of the epidermal growth factor receptor ( P00533 ) tyrosine kinase . We report on an early cellular response to gefitinib that involves induction of functional autophagic flux in phenotypically diverse breast cancer cells that were sensitive ( BT474 and SKBR3 ) or insensitive ( MCF7-GFPLC3 and JIMT-1 ) to gefitinib . Our data show that elevation of autophagy in gefitinib-treated breast cancer cells correlated with downregulation of AKT and P27361 /2 signaling early in the course of treatment . Inhibition of autophagosome formation by BECLIN-1 or O95352 siRNA in combination with gefitinib reduced the abundance of autophagic organelles and sensitized SKBR3 but not MCF7-GFPLC3 cells to cell death . However , inhibition of the late stage of gefitinib-induced autophagy with hydroxychloroquine ( HCQ ) or bafilomycin A1 significantly increased ( p < 0.05 ) cell death in gefitinib-sensitive SKBR3 and BT474 cells , as well as in gefitinib-insensitive JIMT-1 and MCF7-GFPLC3 cells , relative to the effects observed with the respective single agents . Treatment with the combination of gefitinib and HCQ was more effective ( p < 0.05 ) in delaying tumor growth than either monotherapy ( p > 0.05 ) , when compared to vehicle-treated controls . Our results also show that elevated autophagosome content following short-term treatment with gefitinib is a reversible response that ceases upon removal of the drug . In aggregate , these data demonstrate that elevated autophagic flux is an early response to gefitinib and that targeting P00533 and autophagy should be considered when developing new therapeutic strategies for P00533 expressing breast cancers . Gene expression by PBMC in primary sclerosing cholangitis : evidence for dysregulation of immune mediated genes . Primary sclerosing cholangitis ( PSC ) is a chronic disease of the bile ducts characterized by an inflammatory infiltrate and obliterative fibrosis . The precise role of the immune system in the pathogenesis of PSC remains unknown . We used RNA microarray analysis to identify immune-related genes and pathways that are differentially expressed in PSC . Messenger RNA ( mRNA ) from peripheral blood mononuclear cells ( PBMC ) was isolated from both patients with PSC and age and sex matched healthy controls . Samples from 5 PSC patients and 5 controls were analyzed by microarray and based upon rigorous statistical analysis of the data , relevant genes were chosen for confirmation by RT-PCR in 10 PSC patients and 10 controls . Using unsupervised hierarchical clustering , gene expression in PSC was statistically different from our control population . Interestingly , genes within the P60568 receptor beta , P05231 and Q96HU1 Kinase pathways were found to be differently expressed in patients with PSC compared to controls . Further , individual genes , P01375 induced protein 6 ( TNFaip6 ) and membrane-spanning 4-domains , subfamily A ( ms4a ) were found to be upregulated in PSC while similar to Q99717 ( Q99717 ) was downregulated . In conclusion , several immune-related pathways and genes were differentially expressed in PSC compared to control patients , giving further evidence that this disease is systemic and immune-mediated . DB08860 , an P04035 inhibitor , exerts P29474 -independent protective actions against angiotensin II induced cardiovascular remodeling and renal insufficiency . Angiotensin II ( Ang II ) plays a pivotal role in cardiovascular remodeling leading to hypertension , myocardial infarction , and stroke . DB08860 , an P04035 inihibitor , is known to have pleiotropic actions against the development of cardiovascular remodeling . The objectives of this study were to clarify the beneficial effects as well as the mechanism of action of pitavastatin against Ang II-induced organ damage . C57BL6/J mice at 10 weeks of age were infused with Ang II for 2 weeks and were simultaneously administered pitavastatin or a vehicle . DB08860 treatment improved Ang II-induced left ventricular hypertrophy and diastolic dysfunction and attenuated enhancement of cardiac fibrosis , cardiomyocyte hypertrophy , coronary perivascular fibrosis , and medial thickening . Ang II-induced oxidative stress , cardiac TGFbeta-1 expression , and Smad 2/3 phosphorylation were all attenuated by pitavastatin treatment . DB08860 also reduced Ang II-induced cardiac remodeling and diastolic dysfunction in P29474 -/- mice as in wild-type mice . In P29474 -/- mice , the Ang II-induced cardiac oxidative stress and TGF-beta-Smad 2/3 signaling pathway were enhanced , and pitavastatin treatment attenuated the enhanced oxidative stress and the signaling pathway . Moreover , pitavastatin treatment reduced the high mortality rate and improved renal insufficiency in Ang II-treated P29474 -/- mice , with suppression of glomerular oxidative stress and TGF-beta-Smad 2/3 signaling pathway . In conclusion , pitavastatin exerts P29474 -independent protective actions against Ang II-induced cardiovascular remodeling and renal insufficiency through inhibition of the TGF-beta-Smad 2/3 signaling pathway by suppression of oxidative stress . DB08860 , a P04035 inhibitor , blocks vascular smooth muscle cell populated-collagen lattice contraction . Constrictive arterial remodeling plays a major role in lumen narrowing following angioplasty . We investigated the effect of pitavastatin , a 3-hydroxy-3-methylglutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor , on vascular smooth muscle cell ( SMC ) -populated collagen lattice contraction , an in vitro model of vascular contraction . Type I collagen gel contraction by SMCs , which are cultured in collagen gel , was used as a model of vascular remodeling . DB08860 pretreatment inhibited 10 % serum- or platelet-derived growth factor-BB ( PDGF ) -induced SMC-mediated collagen lattice contraction in a concentration-dependent manner . The effect of pitavastatin was prevented by mevalonate or geranylgeranyl pyrophosphate , but not by squalene , a precursor of cholesterol , or farnesyl pyrophosphate . The serum- or PDGF-induced SMC-mediated collagen gel contraction was inhibited by GGTI-298 , a geranylgeranyltransferase inhibitor , P01024 exoenzyme , an inhibitor of Rho , or Y27634 , a Rho kinase inhibitor , but not by FTI-277 , a farnesyltransferase inhibitor . Serum or PDGF treatment increased the stress fiber organization in SMCs , which was blocked by the pitavastatin pretreatment . DB08860 had no effect on the serum- and PDGF-induced lamelliopodia extension of SMC . These results may suggest that pitavastatin attenuates SMC-mediated collagen gel contraction probably via an inhibition of geranylgeranylated Rho protein and a disruption of actin cytoskeletal reorganization . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . Antitumor effect and antiangiogenic potential of the P42345 inhibitor temsirolimus against malignant pleural mesothelioma . The P42345 inhibitor temsirolimus has antitumor and antiangiogenic activity against several carcinomas , yet few reports document the efficacy of temsirolimus against malignant pleural mesothelioma ( MPM ) . Therefore , we evaluated the efficacy of temsirolimus and the antiangiogenic effect of temsirolimus in the treatment of MPM . We examined the efficacy of temsirolimus alone and the efficacy of the combination of temsirolimus and cisplatin or pemetrexed against four MPM cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ( MTT ) assay . The effect of temsirolimus on the production of proangiogenic cytokines by MPM cell lines was examined by enzyme-linked immunosorbent assay ( ELISA ) . Expression of P42345 and proangiogenic cytokines in clinical specimens from MPM patients was determined by immunohistochemistry . DB06287 inhibited cell viability and suppressed cell proliferation of all MPM cell lines . Combined treatment with temsirolimus and cisplatin inhibited the viability of all MPM cell lines more effectively than temsirolimus alone . DB06287 strongly inhibited the phosphorylation of p70s6k , a downstream molecule of P42345 , in all MPM cell lines and led to an increase in the levels of cleaved caspase-3 in the H226 and Y-meso14 cells . DB06287 also inhibited the production of vascular endothelial growth factor ( P15692 ) and platelet-derived growth factor-AA ( PDGF-AA ) . Phosphorylated P42345 and high expression of P15692 and PDGF were detected in 2 and 3 , respectively , out of the 5 MPM specimens . These results suggest that temsirolimus has activity against MPM cells by inhibition of cell proliferation and angiogenesis , and may be beneficial for a subset of MPM patients with high P42345 expression . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . Q07869 gamma ligands , rosiglitazone and pioglitazone , inhibit P09038 - and P15692 -mediated angiogenesis . OBJECTIVE : To study the effect of peroxisome proliferator-activated receptor-gamma ( Q07869 gamma ) agonists , pioglitazone and rosiglitazone , on vascular endothelial growth factor ( P15692 ) - and basic fibroblast growth factor ( P09038 ) -induced angiogenesis and on endothelial cell migration . METHODS : Chick chorioallantoic membrane ( P62158 ) model was used to evaluate the efficacy of pioglitazone and rosiglitazone on P15692 - and P09038 -induced angiogenesis . In addition , the effect of pioglitazone and rosiglitazone on endothelial cell migration was evaluated using 8 mm pore filter to a feeder layer containing vitronectin as chemoattractant . RESULTS : Pioglitazone and rosiglitazone inhibited the pro-angiogenic effects of P09038 and P15692 in the P62158 model significantly ( P < 0.001 ) to the same extent . Endothelial cell migration was also inhibited by both pioglitazone and rosiglitazone ( P < 0.001 ) . CONCLUSIONS : These results suggest that Q07869 gamma ligands , pioglitazone and rosiglitazone , in addition to their important regulatory role in adipogenesis and inflammation , possess anti-angiogenic properties . Thus , Q07869 gamma ligands may be useful in the treatment of diabetic retinopathy , macular degeneration , and other ocular disorders and may lower the risk to develop cancer in diabetic patients . Endothelial cell-derived nitric oxide enhances aerobic glycolysis in astrocytes via HIF-1α-mediated target gene activation . Astrocytes exhibit a prominent glycolytic activity , but whether such a metabolic profile is influenced by intercellular communication is unknown . Treatment of primary cultures of mouse cortical astrocytes with the nitric oxide ( NO ) donor DetaNONOate induced a time-dependent enhancement in the expression of genes encoding various glycolytic enzymes as well as transporters for glucose and lactate . Such an effect was shown to be dependent on the hypoxia-inducible factor HIF-1α , which is stabilized and translocated to the nucleus to exert its transcriptional regulation . NO action was dependent on both the PI3K/Akt/ P42345 and MEK signaling pathways and required the activation of P36551 , but was independent of the soluble guanylate cyclase pathway . Furthermore , as a consequence of NO treatment , an enhanced lactate production and release by astrocytes was evidenced , which was prevented by downregulating HIF-1α . Several brain cell types represent possible sources of NO . It was found that endothelial cells , which express the endothelial NO synthase ( P29474 ) isoform , constitutively produced the largest amount of NO in culture . When astrocytes were cocultured with primary cultures of brain vascular endothelial cells , stabilization of HIF-1α and an enhancement in glucose transporter-1 , hexokinase-2 , and monocarboxylate transporter-4 expression as well as increased lactate production was found in astrocytes . This effect was inhibited by the NOS inhibitor l-NAME and was not seen when astrocytes were cocultured with primary cultures of cortical neurons . Our findings suggest that endothelial cell-derived NO participates to the maintenance of a high glycolytic activity in astrocytes mediated by astrocytic HIF-1α activation . Androgen receptors in the posterior bed nucleus of the stria terminalis increase neuropeptide expression and the stress-induced activation of the paraventricular nucleus of the hypothalamus . The posterior bed nuclei of the stria terminalis ( BST ) are important neural substrate for relaying limbic influences to the paraventricular nucleus ( PVN ) of the hypothalamus to inhibit hypothalamic-pituitary-adrenal ( Q9Y251 ) axis responses to emotional stress . P10275 -expressing cells within the posterior BST have been identified as projecting to the PVN region . To test a role for androgen receptors in the posterior BST to inhibit PVN motor neurons , we compared the effects of the non-aromatizable androgen dihydrotestosterone ( DB02901 ) , the androgen receptor antagonist hydroxyflutamide ( HF ) , or a combination of both drugs implanted unilaterally within the posterior BST . Rats bearing unilateral implants were analyzed for PVN Fos induction in response to acute-restraint stress and relative levels of corticotrophin-releasing hormone and arginine vasopressin ( AVP ) mRNA . DB00142 decarboxylase ( Q99259 ) 65 and Q99259 67 mRNA were analyzed in the posterior BST to test a local involvement of GABA . There were no changes in Q99259 expression to support a GABA-related mechanism in the BST . For PVN neuropeptide expression and Fos responses , basic effects were lateralized to the sides of the PVN ipsilateral to the implants . However , opposite to our expectations of an inhibitory influence of androgen receptors in the posterior BST , PVN AVP mRNA and stress-induced Fos were augmented in response to DB02901 and attenuated in response to HF . These results suggest that a subset of androgen receptor-expressing cells within the posterior BST region may be responsible for increasing the biosynthetic capacity and stress-induced drive of PVN motor neurons . Caenorhabditis elegans expressed sequence tags identify gene families and potential disease gene homologues . A database containing mapped partial cDNA sequences from Caenorhabditis elegans will provide a ready starting point for identifying nematode homologues of important human genes and determining their functions in C. elegans . A total of 720 expressed sequence tags ( ESTs ) have been generated from 585 clones randomly selected from a mixed-stage C. elegans cDNA library . Comparison of these ESTs with sequence databases identified 422 new C. elegans genes , of which 317 are not similar to any sequences in the database . Twenty-six new genes have been mapped by YAC clone hybridization . Members of several gene families , including cuticle collagens , GTP-binding proteins , and RNA helicases were discovered . Many of the new genes are similar to known or potential human disease genes , including P13569 and the P01130 . Silencing urokinase in the ventral tegmental area in vivo induces changes in cocaine-induced hyperlocomotion . DB00133 proteases in the nervous system have functional roles in neural plasticity . Among them , urokinase-type plasminogen activator ( uPA ) exerts a variety of functions during development , and is involved in learning and memory . Furthermore , psychostimulants strongly induce uPA expression in the mesolimbic dopaminergic pathway . In this study , doxycycline-regulatable lentiviruses expressing either uPA , a dominant-negative form of uPA , or non-regulatable lentiviruses expressing small interfering RNAs ( siRNAs ) targeted against uPA have been prepared and injected into the ventral tegmental area ( VTA ) of rat brains . Over-expression of uPA in the VTA induces doxycycline-dependent expression of its receptor , Q03405 , but not its inhibitor , plasminogen activator inhibitor-1 ( P05121 ) . Q03405 expression in the VTA is repressed upon silencing of uPA with lentiviruses expressing siRNAs . In addition , over-expression of uPA in the VTA promotes a 15-fold increase in locomotion activity upon cocaine delivery . Animals expressing the dominant-negative form of uPA did not display such hyperlocomotor activity . These cocaine-induced behavioural changes , associated with uPA expression , could be suppressed in the presence of doxycycline or uPA-specific siRNAs expressing lentiviruses . These data strongly support the major role of urokinase in cocaine-mediated plasticity changes . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Fluorescence imaging reveals the nuclear behavior of peroxisome proliferator-activated receptor/retinoid X receptor heterodimers in the absence and presence of ligand . In a global approach combining fluorescence recovery after photobleaching ( P42345 ) , fluorescence correlation spectroscopy ( FCS ) , and fluorescence resonance energy transfer ( FRET ) , we address the behavior in living cells of the peroxisome proliferator-activated receptors ( PPARs ) , a family of nuclear receptors involved in lipid and glucose metabolism , inflammation control , and wound healing . We first demonstrate that unlike several other nuclear receptors , PPARs do not form speckles upon ligand activation . The subnuclear structures that may be observed under some experimental conditions result from overexpression of the protein and our immunolabeling experiments suggest that these structures are subjected to degradation by the proteasome . Interestingly and in contrast to a general assumption , PPARs readily heterodimerize with retinoid X receptor ( RXR ) in the absence of ligand in living cells . Q07869 diffusion coefficients indicate that all the receptors are engaged in complexes of very high molecular masses and/or interact with relatively immobile nuclear components . PPARs are not immobilized by ligand binding . However , they exhibit a ligand-induced reduction of mobility , probably due to enhanced interactions with cofactors and/or chromatin . Our study draws attention to the limitations and pitfalls of fluorescent chimera imaging and demonstrates the usefulness of the combination of FCS , P42345 , and FRET to assess the behavior of nuclear receptors and their mode of action in living cells .