interacts_with DB00031?
Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Effect of the O00206 antagonist eritoran on retinochoroidal inflammatory damage in a rat model of endotoxin-induced inflammation . PURPOSE : We investigated the effect of eritoran , a O00206 antagonist , on retinochoroidal inflammatory damage in an endotoxin-induced inflammatory rat model . METHODS : Endotoxin-induced inflammatory model was obtained by intraperitoneal injection of 1.5 mg/kg lipopolysaccharide ( LPS ) . Group 1 had control rats ; in groups 2-3 LPS and 0.5 mg/kg sterile saline were injected ; and in groups 4-5 LPS and 0.5 mg/kg eritoran were injected . Blood samples were taken and eyes were enucleated after 12 hours ( h ) ( groups 2 and 4 ) or 24 hours ( Groups 3 and 5 ) . P01375 -α ( P01375 -α ) and malondialdehyde ( MDA ) levels in the serum and retinochoroidal tissue and nuclear factor kappa-B ( NFκB ) levels in retinochoroidal tissue were determined . Histopathological examination was performed and retinochoroidal changes were scored . RESULTS : DB04933 treatment resulted in lower levels of P01375 -α , MDA , and NFκB after 12 h which became significant after 24 h . Serum P01375 -α and retinochoroidal tissue NFκB levels were similar to control animals at the 24th h of the study . DB04933 significantly reversed histopathological damage after 24 h . CONCLUSIONS : DB04933 treatment resulted in less inflammatory damage in terms of serum and retinochoroidal tissue parameters . The retinoid anticancer signal : mechanisms of target gene regulation . Retinoids induce growth arrest , differentiation , and cell death in many cancer cell types . One factor determining the sensitivity or resistance to the retinoid anticancer signal is the transcriptional response of retinoid-regulated target genes in cancer cells . We used cDNA microarray to identify 31 retinoid-regulated target genes shared by two retinoid-sensitive neuroblastoma cell lines , and then sought to determine the relevance of the target gene responses to the retinoid anticancer signal . The pattern of retinoid responsiveness for six of 13 target genes ( RARbeta2 , O43174 , P09455 , O15492 , Q16828 , P18146 ) correlated with phenotypic retinoid sensitivity , across a panel of retinoid-sensitive or -resistant lung and breast cancer cell lines . Retinoid treatment of P04198 transgenic mice bearing neuroblastoma altered the expression of five of nine target genes examined ( RARbeta2 , O43174 , P09455 , Q16828 , P00750 ) in neuroblastoma tumour tissue in vivo . In retinoid-sensitive neuroblastoma , lung and breast cancer cell lines , direct inhibition of retinoid-induced RARbeta2 expression blocked induction of only one of eight retinoid target genes ( O43174 ) . DNA demethylation , histone acetylation , and exogenous overexpression of RARbeta2 partially restored retinoid-responsive O43174 expression in RA-resistant MDA-MB-231 breast , but not SK-MES-1 lung , cancer cells . Combined , rather than individual , inhibition of Q16828 and O15492 was required to block retinoid-induced growth inhibition in neuroblastoma cells , through phosphorylation of extracellular-signal-regulated kinase . In conclusion , sensitivity to the retinoid anticancer signal is determined in part by the transcriptional response of key retinoid-regulated target genes , such as RARbeta2 , Q16828 , and O15492 . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . Surgical stress-induced transient nephrogenic diabetes insipidus ( NDI ) associated with decreased DB00067 receptor2 ( P30518 ) expression linked to nonsense-mediated mRNA decay and incomplete skewed X-inactivation in a female patient with a heterozygous P30518 mutation ( c. 89-90 delAC ) . Induction of cytokine gene expression in human thyroid epithelial cells irradiated with HZE particles ( iron ions ) . Gene expression profiles were examined using cDNA microarray technology in human thyroid epithelial ( Htori-3 ) cells exposed to a low , non-toxic dose ( 10 cGy ) of radiation from HZE particles in the form of iron ions in the absence or presence of selenomethionine ( SeM ) . A total of 215 genes were differentially regulated 2 h after exposure to a 10-cGy dose of iron-ion radiation . In the microarray analysis , SeM had profound effects on the radiation-induced expression of several specific genes , which includes P00749 , P17936 , P15328 , P15291 and P02452 . Of particular interest to us was a gene cluster , " secreted proteins " , that was up-regulated after radiation exposure . Seven up-regulated genes of this gene cluster fall within the chemokine/cytokine gene cluster , namely , P09341 , P19875 , P05231 , P20809 , P10145 , Q13007 and TGFbeta2 . In microarray studies , the radiation-induced up-regulated expression of some these genes encoding cytokine/chemokine proteins was significantly decreased by SeM treatment . For P10145 , TGFbeta2 , P09341 and P19875 , these observations were validated by qPCR techniques . It is concluded that SeM can regulate ionizing radiation-induced gene expression and may serve as an effective countermeasure for some of the acute inflammatory/immune responses induced by low-dose HZE-particle radiation . [ DB00162 transport into the cell nucleus in vitro ] . The retinol interaction with intact cellular nuclei , nuclear envelope and chromatin was investigated . We have shown that the Cellular DB00162 -Binding Protein ( P09455 ) plays a very important role in such interaction . DB00162 can specifically interacts with nuclei , nuclear envelope and chromatin only when it presents as a complex with P09455 . The obtained data allowed us to suggest that the process of retinol delivery to hypothetical nuclear receptors include of at least the following stages : 1 ) specific binding with nuclear envelope ; 2 ) penetration into nuclei ; 3 ) specific binding with chromatin receptors . P09455 is a necessary component at all this stages . Also we show that P09455 has not a species or tissue specificity . DB00067 -induced P04275 secretion from endothelial cells involves V2 receptors and DB02527 . DB00067 and its analogue DB00035 ( DB00035 ) are known to raise plasma P04275 ( P04275 ) levels . DB00035 is used as a hemostatic agent for the treatment of von Willebrand 's disease . However , its cellular mechanisms of action have not been elucidated . DB00035 , a specific agonist for the vasopressin V2 receptor ( P30518 ) , exerts its antidiuretic effect via a rise in DB02527 in kidney collecting ducts . We tested the hypothesis that DB00035 induces P04275 secretion by binding to P30518 and activating DB02527 -mediated signaling in endothelial cells . P04275 secretion from human umbilical vein endothelial cells ( HUVECs ) can be mediated by DB02527 , but DB00035 is ineffective , presumably due to the absence of P30518 . We report that DB00035 stimulates P04275 secretion in a DB02527 -dependent manner in HUVECs after transfection of the P30518 . In addition , vasopressin and DB00035 induce P04275 secretion in human lung microvascular endothelial cells ( HMVEC-L ) . These cells ( but not HUVECs ) express endogenous P30518 , as shown by RT-PCR . DB00067 -induced P04275 secretion is mimicked by DB00035 and inhibited by the selective P30518 antagonist SR121463B . It is mediated by DB02527 , since it is inhibited by the protein kinase A inhibitor Rp-8CPT-cAMPS . These results indicate that vasopressin induces DB02527 -mediated P04275 secretion by a direct effect on endothelial cells . They also demonstrate functional expression of P30518 in endothelial cells , and provide a cellular mechanism for the hemostatic effects of DB00035 . Modulation of proteolytic activity during neuritogenesis in the PC12 nerve cell : differential control of plasminogen activator and plasminogen activator inhibitor activities by nerve growth factor and dibutyryl-cyclic AMP . Extracellular proteolysis is considered to be required during neuritic outgrowth to control the adhesiveness between the growing neurite membrane and extracellular matrix proteins . In this work , PC12 nerve cells were used to study the modulation of proteolytic activity during neuronal differentiation . PC12 cells were found to contain and release a 70-75-kDa tissue-type plasminogen activator ( tPA ) and a much less abundant 48-kDa urokinase-type plasminogen activator . A plasminogen activator inhibitor ( P05121 ) activity with molecular sizes of 54 and 58 kDa was also detected in PC12 cell conditioned medium and formed high-molecular-mass complexes with released tPA . Release of P05121 activity was dependent on treatment with nerve growth factor ( P01138 ) , whereas tPA synthesis and release were under control of a cyclic AMP-dependent mechanism and increased on treatment with dibutyryl-cyclic AMP [ (But)2cAMP ] or cholera toxin . Simultaneous treatment with P01138 and (But)2cAMP resulted in increases of both tPA and P05121 release and enhancement of tPA- P05121 complex formation . The resulting plasminogen activator activity in conditioned medium was high in (But)2cAMP-treated cultures with short neuritic outgrowth but remained low in P01138 - or P01138 plus (But)2cAMP-treated cultures , where neurite extension was , respectively , large and very large . These results suggest that excess proteolytic activity may be detrimental to neuritic outgrowth and that not only P05121 release but also tPA- P05121 complex formation is associated with production of large and stable neuritic outgrowth . This can be understood as an involvement of P05121 in the protection against neurite-destabilizing proteolytic activity . Predictive value of circulating interleukin-6 and heart-type fatty acid binding protein for three months clinical outcome in acute cerebral infarction : multiple blood markers profiling study . INTRODUCTION : There is no single blood marker for predicting the prognosis in ischemic stroke . A combination of multiple blood markers may enhance the ability to predict long-term outcome following ischemic stroke . METHODS : Blood concentrations of neuronal markers ( neuron-specific enolase , visinin-like protein 1 , heart type fatty acid binding protein ( hFABP ) and neuroglobin ) , astroglial markers ( P04271 and glial fibrillary acidic protein ) , inflammatory markers ( P05231 , P01375 -α , and P02741 ) , blood-brain barrier marker ( matrix metalloproteinase 9 ) , and haemostatic markers ( D-dimer and P05121 ) were measured within 24 hours after stroke onset . The discrimination and reclassification for favorable and poor outcome were compared after adding individual or a combination of blood markers to the clinical model of stroke outcome . RESULTS : In multivariate analysis , natural log-transformed ( log ) P05231 ( odds ratio ( OR ) : 1.75 , 95 % CI : 1.25 to 2.25 , P=0.001 ) and loghFABP ( OR : 3.23 , 95 % CI : 1.44 to 7.27 , P=0.005 ) were independently associated with poor outcome . The addition of a single blood marker to the clinical model did not improve the discriminating ability of the clinical model of stroke outcome . However , the addition of the combination of logIL-6 and loghFABP to the clinical model showed improved discrimination ( area under receiver operating characteristic ( AUROC ) curve : 0.939 versus 0.910 , P=0.03 ) and reclassification performance ( net reclassification improvement index : 0.18 , P=0.005 ) . CONCLUSIONS : A combination of circulating P05231 and hFABP level has an additive clinical value for the prediction of stroke outcome . DB02342 causes functional repression of transforming growth factor β3 signaling by ameliorating Smad and non-Smad signaling pathways in immortalized uterine fibroid cells . OBJECTIVE : To investigate the effects and the mechanism of action of 2-methoxyestradiol ( 2ME(2) ) on transforming growth factor ( TGF ) β3-induced profibrotic response in immortalized human uterine fibroid smooth muscle ( huLM ) cells . DESIGN : Laboratory study . SETTING : University research laboratory . PATIENTS(S) : Not applicable . INTERVENTIONS(S) : Not applicable . MAIN OUTCOME MEASURE(S) : huLM cells were treated with TGF-β3 ( 5 ηg/mL ) in the presence or absence of specific P84022 inhibitor SIS3 ( 1 μmol/L ) , inhibitor of the PI3K/Akt ( LY294002 , 10 μmol/L ) , or 2ME(2) ( 0.5 μmol/L ) , and the expression of collagen ( Col ) type I(αI) , Col III(αI) , plasminogen activator inhibitor ( P05121 ) 1 , connective tissue growth factor ( P29279 ) , and α-smooth muscle actin ( α-SMA ) were determined by real-time reverse-transcription polymerase chain reaction and immunoblotting . The effect of 2ME(2) on Smad-microtubule binding was evaluated by coimmunoprecipitation . RESULT(S) : Our data revealed that TGF-β3-induced fibrogenic response in huLM is mediated by both Smad-dependent and Smad-independent PI3K/Akt/ P42345 signaling pathways . 2ME(2) abrogates TGF-β3-induced expression of Col I(αI) , Col III(αI) , P05121 , P29279 , and α-SMA . Molecularly , 2ME(2) ameliorates TGF-β3-induced Q15796 /3 phosphorylation and nuclear translocation . In addition , 2ME(2) inhibits TGF-β3-induced activation of the PI3K/Akt/ P42345 pathway . CONCLUSION(S) : TGF-β3-induced profibrotic response in fibroid cells is mediated by Smad-dependent and Smad-independent PI3K/Akt/ P42345 pathways . 2ME(2) inhibits TGF-β3 profibrotic effects in huLM cells by ameliorating both Smad-dependent and Smad-independent signaling pathways . P0DMS8 is a critical mediator in LPS-induced pulmonary inflammation . DB00640 receptor A(3) ( A(3) ) regulates directed movement of polymorphonuclear cells ( PMNs ) to sites of inflammation and has been implicated as a relevant mediator in models of inflammatory diseases . Here , we sought to characterize the role of A(3) in a murine model of lung inflammation . Initial studies revealed that pulmonary A(3) transcript levels were elevated following LPS exposure in vivo . In addition , inhalation of LPS increased the accumulation of PMNs in wild-type and A(3)(-/-) mice in all lung compartments . Pretreatment with the specific A(3)-agonist Cl- DB05511 significantly decreased migration of PMNs into lung interstitium and alveolar air space of wild-type mice but not of A(3)(-/-) mice . Lower PMN counts were associated with reduced levels of P01375 -α and P05231 in the alveolar space of wild-type mice that received Cl- DB05511 . In addition , Cl- DB05511 attenuated LPS-induced microvascular permeability in wild-type mice as assessed by the extravasation of Evans blue . In pulmonary microvascular endothelial cells , Cl- DB05511 reduced LPS-induced cytoskeletal remodeling and cell retraction , consistent with a specific role of A(3) for maintaining endothelial integrity . Migratory activity of human PMNs across an endothelial or epithelial monolayer was reduced when A(3) was activated on PMNs . Studies in chimeric mice , however , revealed that Cl- DB05511 required A(3) on both hematopoietic and nonhematopoietic cells to reduce transmigration in vivo . Together , our results shed new light on the role of A(3) in LPS-induced PMN trafficking in the lung and suggest pharmacological modulation of A(3)-dependent pathways as a promising approach in lung inflammation . DB00054 : a reappraisal of its use in coronary care . Platelet reactivity plays a pivotal role in the pathogenesis of ischemic adverse events during and after acute coronary syndromes ( ACS ) , and percutaneous coronary intervention ( P05154 ) . Glycoprotein ( GP ) IIb/IIIa inhibitors are the strongest antiplatelet agents currently available on the market and three different compounds , namely abciximab , tirofiban , and eptifibatide , have been approved for clinical use . DB00054 has been investigated in the clinical field far more extensively than the other P08514 /IIIa inhibitors . DB00054 is an anti-integrin Fab fragment of a human - mouse chimeric monoclonal antibody with high affinity and a slow dissociation rate from the GP IIb/IIIa platelet receptor . DB00054 , given shortly before the coronary intervention , is superior to placebo in reducing the acute risk of ischemic complications ( EPIC , EPISTENT , EPILOG trials ) ; moreover , in the ISAR-REACT 2 study abciximab has been shown to reduce the risk of adverse events in patients with non ST-segment elevation ACS who are undergoing P05154 even after optimal pre-treatment with 600 mg of clopidogrel . Finally , abciximab has been also used in abciximab-coated stent , with only bolus administration regimen and for direct intracoronary use with promising results that may extend and/or modify its current use in clinical practice in future . Ca2+ or Sr2+ partially rescues synaptic transmission in hippocampal cultures treated with botulinum toxin A and C , but not tetanus toxin . Botulinum ( DB00083 -G ) and tetanus toxins ( TeNT ) are zinc endopeptidases that cleave proteins associated with presynaptic terminals ( P60880 , syntaxin , or VAMP/synaptobrevin ) and block neurotransmitter release . Treatment of hippocampal slice cultures with DB00083 , BoNT/C , BoNT/E , or TeNT prevented the occurrence of spontaneous or miniature EPSCs ( sEPSCs or mEPSCs ) as well as the [Ca2+]o-independent increase in their frequency induced by phorbol ester , 0.5 nM alpha-latrotoxin , or sucrose . [Ca2+]o-independent and -dependent release thus requires that the target proteins of clostridial neurotoxins be uncleaved . In contrast , significant increases in mEPSC frequency were produced in BoNT-treated , but not TeNT-treated , cultures by application of the Ca2+ ionophore ionomycin in the presence of 10 mM [Ca2+]o . The frequency of sEPSCs was increased in BoNT-treated , but not TeNT-treated , cultures by increasing [Ca2+]o from 2.8 to 5-10 mM or by applying 5 mM Sr2+ . Large Ca2+ and Sr2+ influxes thus can rescue release after BoNT treatment , albeit less than in control cultures . The nature of the toxin-induced modification of Ca2+-dependent release was assessed by recordings from monosynaptically coupled P07451 cell pairs . The paired-pulse ratio of unitary EPSCs evoked by two presynaptic action potentials in close succession was 0.5 in control cultures , but it was 1.4 and 1.2 in DB00083 - or BoNT/C-treated cultures when recorded in 10 mM [Ca2+]o . Log-log plots of unitary EPSC amplitude versus [Ca2+]o were shifted toward higher [Ca2+]o in DB00083 - or BoNT/C-treated cultures , but their slope was unchanged and the maximal EPSC amplitudes were reduced . We conclude that BoNTs reduce the Ca2+ sensitivity of the exocytotic machinery and the number of quanta released . Neuroprotective effects of huperzine A : new therapeutic targets for neurodegenerative disease . In recent years , the most common pharmacological treatment for Alzheimer 's disease ( AD ) has been acetylcholinesterase ( P22303 ) inhibition . However , this single-target approach has limited effectiveness and there is evidence that a multitarget approach might be more effective . DB04864 ( HupA ) , a novel alkaloid isolated from a Chinese herb , has neuroprotective effects that go beyond the inhibition of P22303 . Recent data have demonstrated that HupA can ameliorate the learning and memory deficiency in animal models and AD patients . Its potentially beneficial actions include modification of beta-amyloid peptide processing , reduction of oxidative stress , neuronal protection against apoptosis , and regulation of the expression and secretion of nerve growth factor ( P01138 ) and P01138 signaling . Regulation by nitric oxide of endotoxin-induced tissue factor and plasminogen activator inhibitor-1 in endothelial cells . The increase in nitric oxide ( NO ) production in lipopolysaccharide ( LPS ) -induced sepsis is thought to contribute to the development of shock . However , NO could also play an antithrombotic role . Little is known about the modulating effect of NO on the endothelial overexpression and production of tissue factor ( TF ) and plasminogen activator inhibitor-1 ( P05121 ) occurring in endotoxemia . We analyzed the effect of N(G)-nitro-L-arginine-methyl-ester ( L-NAME ) , an inhibitor of NO synthases , and S-nitroso-N-acetyl-D,L-penicillamine ( P60880 ) , a NO donor , on the expression and synthesis of TF and P05121 by LPS-challenged human umbilical vein endothelial cells ( HUVEC ) : L-NAME enhanced the increase in TF mRNA and antigen levels ( P < 0.05 ) observed in LPS-treated HUVEC ; P60880 down-regulated the LPS-induced TF increment ( p < 0.05 ) . However , no effects of NO on regulation of the LPS-dependent increase in P05121 could be seen . Thus , NO could play an antithrombotic role in sepsis by down-regulating the endothelial overexpression and production of TF . 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 . DB05595 , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC(50) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 did not block membrane binding of radiolabeled folic acid , 5-methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 had a minimal effect on the cytoplasmic accumulation of 5-methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC(50) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity .